Label-free microfluidic chip for segregation and recovery of circulating leukemia cells: clinical applications in acute myeloid leukemia.
We present a label-free microfluidic chip for the segregation of circulating leukemia cells (CLCs) from blood samples, with a focus on its clinical applications in Acute Myeloid Leukemia (AML). The microfluidic chip achieved an approximate capture efficiency of 92%. The study analyzed a comprehensive set of 66 blood specimens from AML patients in different disease stages, including newly diagnosed and relapsing cases, patients in complete remission, and those in partial remission. The results showed a significant difference in CLC counts between active disease stages and remission stages (p < 0.0001), with a proposed threshold of 5 CLCs to differentiate between the two. The microfluidic chip exhibited a sensitivity of 95.4% and specificity of 100% in predicting disease recurrence. Additionally, the captured CLCs were subjected to downstream molecular analysis using droplet digital PCR, allowing for the identification of genetic mutations associated with AML. Comparative analysis with bone marrow aspirate processing by FACS demonstrated the reliability and accuracy of the microfluidic chip in tracking disease burden, with highly agreement results obtained between the two methods. The non-invasive nature of the microfluidic chip and its ability to provide real-time insights into disease progression make it a promising tool for the proactive monitoring and personalized patient care of AML.


[Diabetes mellitus and atypical antipsychotics].
Schizophrenic patients present a higher risk for the development of hyperglycemic disorder and the use of antipsychotic drugs seems to increase the risk of diabetes mellitus. The present review concerns the relation between atypical antipsychotic drugs and the risk of developing diabetes mellitus. A Medline and Webofscience search was performed by using the terms "Hyperglycemia", "Diabetes Mellitus" and "Antipsychotic Agents", to identify original papers and reviews published between 1997 and september 2002. It is concluded that there is a higher risk of glycemic disorders in the population of patients treated by antipsychotic drugs. Dietetic measures and attention to risk factors should be taken into account during the treatment of psychotic patients.


Affibody-mediated transferrin depletion for proteomics applications.
An Affibody (Affibody) ligand with specific binding to human transferrin was selected by phage display technology from a combinatorial protein library based on the staphylococcal protein A (SpA)-derived Z domain. Strong and selective binding of the selected Affibody ligand to transferrin was demonstrated using biosensor technology and dot blot analysis. Impressive specificity was demonstrated as transferrin was the only protein recovered by affinity chromatography from human plasma. Efficient Affibody-mediated capture of transferrin, combined with IgG- and HSA-depletion, was demonstrated for human plasma and cerebrospinal fluid (CSF). For plasma, 85% of the total transferrin content in the samples was depleted after only two cycles of transferrin removal, and for CSF, 78% efficiency was obtained in single-step depletion. These results clearly suggest a potential for the development of Affibody-based resins for the removal of abundant proteins in proteomics analyses.


Breath biomarkers in diagnosis of pulmonary diseases.
Breath analysis provides a convenient and simple alternative to traditional specimen testing in clinical laboratory diagnosis. As such, substantial research has been devoted to the analysis and identification of breath biomarkers. Development of new analytes enhances the desirability of breath analysis especially for patients who monitor daily biochemical parameters. Elucidating the physiologic significance of volatile substances in breath is essential for clinical use. This review describes the use of breath biomarkers in diagnosis of asthma, chronic obstructive pulmonary disease (COPD), cystic fibrosis (CF), lung cancer, as well as other pulmonary diseases. A number of breath biomarkers in lung pathophysiology will be described including nitric oxide (NO), carbon monoxide (CO), hydrogen peroxide (H₂O₂) and other hydrocarbons.


On the Security of a Class of Diffusion Mechanisms for Image Encryption.
The need for fast and strong image cryptosystems motivates researchers to develop new techniques to apply traditional cryptographic primitives in order to exploit the intrinsic features of digital images. One of the most popular and mature technique is the use of complex dynamic phenomena, including chaotic orbits and quantum walks, to generate the required key stream. In this paper, under the assumption of plaintext attacks we investigate the security of a classic diffusion mechanism (and of its variants) used as the core cryptographic primitive in some image cryptosystems based on the aforementioned complex dynamic phenomena. We have theoretically found that regardless of the key schedule process, the data complexity for recovering each element of the equivalent secret key from these diffusion mechanisms is only (1). The proposed analysis is validated by means of numerical examples. Some additional cryptographic applications of this paper are also discussed.


Nonoperative Management of Appendicitis.
We evaluated the outcomes for nonoperative management (NOM) of all children with suspected nonperforated appendicitis, including those patients with an appendicolith. Parents of all children with suspected nonperforated appendicitis were offered NOM versus laparoscopic appendectomy. NOM included administration of intravenous antibiotics and hospital admission. If no improvement within 24 hours, laparoscopic appendectomy was performed. Primary outcomes were initial success rate and recurrence rate. Fifty patients selected NOM. The initial failure rate for NOM was 20%. Of the 10 who failed, 7 had complicated appendicitis. The recurrence rate was 13%. Overall, 34 (68%) patients avoided appendectomy. Patients with an appendicolith had a higher initial failure rate (37%) compared to patients without an appendicolith (10%; P < .05). NOM is feasible and effective in pediatric nonperforated appendicitis. The presence of an appendicolith was associated with a higher failure rate but is not an absolute contraindication for NOM.


[3H]WB4101 labels the 5-HT1A serotonin receptor subtype in rat brain. Guanine nucleotide and divalent cation sensitivity.
In the presence of a 30 nM prazosin mask, [3H]-2-(2,6-dimethoxyphenoxyethyl) aminomethyl-1,4-benzodioxane ([3H]WB4101) can selectively label 5-HT1 serotonin receptors. Serotonin exhibits high affinity (Ki = 2.5 nM) and monophasic competition for [3H] WB4101 binding in cerebral cortex. Furthermore, we have found a significant correlation (r = 0.96) between the affinities of a number of serotonergic and nonserotonergic compounds at [3H]WB4101-binding sites in the presence of 30 nM prazosin and [3H] lysergic acid diethylamide ([3H]LSD)-labeled 5-HT1 serotonin receptors in homogenates of rat cerebral cortex. Despite similar pharmacological profiles, distribution studies indicate that, in the presence of 5 mM MgSO4, the Bmax of [3H]WB4101 is significantly lower than the Bmax of [3H]LSD in various brain regions. WB4101 competition for [3H] LSD-labeled 5-HT1 receptors fits best to a computer-derived model assuming two binding sites, with the KH for WB4101 being similar to the KD of [3H]WB4101 binding derived from saturation experiments. This suggests that [3H]WB4101 labels only one of the subtypes of the 5-HT1 serotonin receptors labeled by [3H]LSD. Interestingly, the selective 5-HT1A serotonin receptor antagonist, spiperone, and the selective 5-HT1A agonist, 8-hydroxy-2-(di-n-propylamino) tetraline, exhibit high affinity and monophasic competition for [3H]WB4101 but compete for multiple [3H]LSD 5-HT1 binding sites. These data indicate that [3H]WB4101 selectively labels the 5-HT1A serotonin receptor, whereas [3H] LSD appears to label both the 5-HT1A and the 5-HT1B serotonin receptor subtypes. The divalent cations, Mn2+, Mg2+, and Ca2+ were found to markedly increase the affinity and Bmax of [3H]WB4101 binding in cerebral cortex. Conversely, the guanine nucleotides guanylylimidodiphosphate and GTP, but not the adenosine nucleotide ATP, markedly reduce the Bmax of [3H]WB4101 binding. These characteristics are typical of agonists interacting with receptors which modulate cellular function via a guanine nucleotide-regulatory subunit.


Nimodipine modulates Bcl-2 and Bax mRNA expression after cerebral ischemia.
In order to explore whether the member of Bcl-2 gene family, for example, Bcl-2 and Bax, are induced after cerebral ischemia, and whether expression of genes can be modulated by calcium-antagonist, the rat cerebral ischemic models were made by occluding left middle cerebral artery. The expression of Bcl-2 and Bax mRNA was measured by RT-PCR method. After middle cerebral artery occlusion (MCAO), the expression of both Bcl-2 and Bax mRNA were induced. Level of Bcl-2 mRNA increased steadily and level of Bax mRNA increased gradually at first, reached a peak after 24 h, then decreased slowly. After administration of nimodipine, Bcl-2 mRNA was up-regulated in the hippocampus 6 and 24 h after ischemia, while Bax mRNA was down-regulated 6 and 24 h after ischemia. Focal cerebral ischemia can induce proto-oncogenes to express, which was associated with apoptosis. Calcium-antagonist can up-regulate Bcl-2 mRNA and down-regulate Bax mRNA. The increased ratio of Bcl-2 and Bax mRNA may contribute to the anti-apoptic effect of nimodipine. The study indicates that pharmacological modulation of Bcl-2 family member expression could become a new strategy to manage neuronal damage.


Identifying patient-centered quality indicators for the care of adult home parenteral nutrition (HPN) patients.
Previous qualitative research, aimed at eliciting the experiences and views of home parenteral nutrition (HPN) patients in daily life, revealed little information on HPN care. However, to perform patient-centered care, it is important to understand what "quality of care" means for adult HPN patients. Therefore, the aim of this study was to identify quality indicators through patients' perspectives. In-depth, face-to-face, semistructured interviews were conducted between June and November 2011 with adult patients who recently started HPN. Patients were selected by purposive sampling. Interviews were audio recorded, transcribed verbatim, and analyzed using a framework approach. Nvivo 9.0 software was used for managing transcripts. Twelve adult patients (7F/5M) were interviewed. At the time of the interview, median HPN experience was 59 days. Two structure, 31 process, and 9 outcome indicators were identified. The majority of process indicators are related to the home nurse, indicating the important role of these healthcare professionals in HPN care. In addition, all interviewees reported communication as an essential aspect of the complex care pathway. The identified outcome indicators are comparable with the indicators that were previously developed by an expert panel. For example, improvement in quality of life during HPN treatment and incidence of catheter-related infections were identified by both groups. Interviews with HPN patients revealed an interesting set of indicators that can be used to perform, evaluate, or improve current HPN care and are a starting point for a more patient-centered approach.


The association between thyroid volume, L-thyroxine therapy and hepatocyte growth factor levels among patients with euthyroid and hypothyroid goitrous and non-goitrous Hashimoto's thyroiditis versus healthy subjects.
Hashimoto's thyroiditis (HT) is the most common etiology of hypothyroidism in regions where iodine deficiency is not a concern. To date, many clinical investigations have been conducted to elucidate its pathogenesis. Several growth factors have been shown to have a role in its development. Hepatocyte growth factor (HGF) is one of the aforementioned molecules. We aimed to demonstrate whether HGF is responsible for HT and goiter development. Also, we aimed to test the hypothesis that levo-thyroxine sodium therapy will suppress HGF levels. Sixty-one premenopausal women who were admitted to our outpatient clinic between November 2010 and September 2011 were enrolled. Three groups were determined according to their thyroid function tests (TFTs) as euthyroid Hashimoto's, control and subclinical hypothyroid Hashimoto's groups. Basal TFTs, anti-thyroid peroxidase (anti-TPO), anti-thyroglobulin (anti-tg), thyroid ultrasonography (USG) and HGF were studied and recorded. Subclinical hypothyroid HT patients received levo-thyroxine sodium replacement therapy, and were re-assessed for the same laboratory and radiologic features after a median 3.5 month follow-up. Basal HGF levels were not different between groups. In the subclinical hypothyroidism group, HGF levels (752.75 ± 144.91 pg/ml vs. 719.37 ± 128.05 pg/ml; p = 0.496) and thyroid volumes (12.51 ± 3.67 cc vs. 12.18 ± 4.26 cc; p = 0.7) before and after treatment did not change significantly. No correlations were found between HGF and other parameters. HGF levels were similar between subjects with nodular goiter and normal thyroid structure. HGF was not shown to be associated with HT and goiter development. In addition, levo-thyroxine sodium replacement therapy did not alter serum HGF levels significantly.


Comparative safety of serotonin (5-HT3) receptor antagonists in patients undergoing surgery: a systematic review and network meta-analysis.
Serotonin (5-HT3) receptor antagonists are commonly used to decrease nausea and vomiting for surgery patients, but these agents may be harmful. We conducted a systematic review on the comparative safety of 5-HT3 receptor antagonists. Searches were done in MEDLINE, Embase, and the Cochrane Central Register of Controlled Trials to identify studies comparing 5-HT3 receptor antagonists with each other, placebo, and/or other antiemetic agents for patients undergoing surgical procedures. Screening search results, data abstraction, and risk of bias assessment were conducted by two reviewers independently. Random-effects pairwise meta-analysis and network meta-analysis (NMA) were conducted. PROSPERO registry number: CRD42013003564. Overall, 120 studies and 27,787 patients were included after screening of 7,608 citations and 1,014 full-text articles. Significantly more patients receiving granisetron plus dexamethasone experienced an arrhythmia relative to placebo (odds ratio (OR) 2.96, 95 % confidence interval (CI) 1.11-7.94), ondansetron (OR 3.23, 95 % CI 1.17-8.95), dolasetron (OR 4.37, 95 % CI 1.51-12.62), tropisetron (OR 3.27, 95 % CI 1.02-10.43), and ondansetron plus dexamethasone (OR 5.75, 95 % CI 1.71-19.34) in a NMA including 31 randomized clinical trials (RCTs) and 6,623 patients of all ages. No statistically significant differences in delirium frequency were observed across all treatment comparisons in a NMA including 18 RCTs and 3,652 patients. Granisetron plus dexamethasone increases the risk of arrhythmia.


Injection 3D Concrete Printing (I3DCP): Basic Principles and Case Studies.
Today, the majority of research in 3D concrete printing focuses on one of the three methods: firstly, material extrusion; secondly, particle-bed binding; and thirdly, material jetting. Common to all these technologies is that the material is applied in horizontal layers. In this paper, a novel 3D concrete printing technology is presented which challenges this principle: the so-called Injection 3D Concrete Printing (I3DCP) technology is based on the concept that a fluid material (M1) is robotically injected into a material (M2) with specific rheological properties, causing material M1 to maintain a stable position within material M2. Different to the layered deposition of horizontal strands, intricate concrete structures can be created through printing spatially free trajectories, that are unconstrained by gravitational forces during printing. In this paper, three versions of this method were investigated, described, and evaluated for their potential in construction: A) injecting a fine grain concrete into a non-hardening suspension; B) injecting a non-hardening suspension into a fine grain concrete; and C) injecting a fine grain concrete with specific properties into a fine grain concrete with different properties. In an interdisciplinary research approach, various material combinations were developed and validated through physical experiments. For each of the three versions, first architectural applications were developed and functional prototypes were fabricated. These initial results confirmed both the technological and economic feasibility of the I3DCP process, and demonstrate the potential to further expand the scope of this novel technology.


Flow-dependent Smad2 phosphorylation and TGIF nuclear localization in human aortic endothelial cells.
Endothelial cells respond to fluid flow stimulation through transient and sustained signal pathway activation. Smad2 is a signaling molecule and transcription factor in the Smad signaling pathway, traditionally associated with TGF-β. Although phosphorylation of Smad2 in the receptor-dependent COOH-terminal region is the most appreciated way Smad2 is activated to affect gene expression, phosphorylation may also occur in the MH1-MH2 linker region (L-psmad2). Here, we show that in human aortic endothelial cells (HAEC), Smad2 was both preferentially phosphorylated in the linker region and localized to the nucleus in a flow-dependent manner. The Smad corepressor transforming growth interacting factor (TGIF) was also found to have flow-dependent nuclear localization. Tissue studies confirmed this L-psmad2 generation trend in rat aorta, indicating likely importance in arterial tissue. HAEC-based inhibitor studies demonstrated that L-psmad2 levels were not related to MAPK phosphorylation, but instead followed the pattern of pAkt(473), both with and without the phosphatidylinositol 3-kinase inhibitor PI-103. Akt and Smad species were also shown to directly interact under flow relative to static controls. To further evaluate impacts of PI-103 treatment, expression profiles for two TGF-β and shear stress-dependent genes were determined and showed that mRNAs were lower from untreated 10 dyn/cm(2) than 2 dyn/cm(2) average shear stress cultures. However, upon exposure to PI-103, this trend was reversed, with a stronger response observed at 10 dyn/cm(2). Taken together, the results of this work suggest that fluid flow exposure may influence endothelial gene expression by a novel mechanism involving Akt, L-psmad2, and TGIF.


Emerging relationship between hydrogen sulfide and ferroptosis: A literature review.
Gaseous hydrogen sulfide (H2S) can function as a signaling molecule similar to nitric oxide or carbon monoxide under physiological conditions, ultimately exerting anti-inflammatory, anti-apoptotic, and antioxidant activities. Many studies have investigated the role of H2S in a variety of biological contexts, and both endogenous H2S and H2S donors have been leveraged as tools for fundamental biomedical research, and it has been suggested that they may provide value for the design of novel therapeutic strategies in the years to come. Ferroptotic cell death is a distinct programmed cell death resulting from excessive lipid peroxidation in an iron-dependent manner, and is characterized by high levels of iron accumulation, reactive oxygen species (ROS) production, and peroxidation of cellular lipids. Several recent studies have revealed a close relationship between ferroproteins and their precursors, H2S, and the enzymes that produce them. This review summarizes the current information pertaining to the relationship between ferroptosis and H2S, with a particular focus on the underlying mechanisms and biological applications of this knowledge.


Decreased frequency of FBN1 missense variants in Ghent criteria-positive Marfan syndrome and characterization of novel FBN1 variants.
The diagnosis of Marfan syndrome (MFS) remains challenging despite the 2010 revision to Ghent nosology criteria, and there is a lack of published information regarding FBN1 genotype associations in patients since the update in Ghent criteria. Applying revised Ghent criteria, we reviewed consecutive proband cases (n=292) submitted for FBN1 sequencing. Testing yielded 207 pathogenic or likely pathogenic FBN1 variants, with 114/207 (55%) missense, 67/207 (32%) non-sense or frameshift, and 28/207 (13%) splicing. There were 130 novel FBN1 variants predicted as pathogenic or likely pathogenic (n=109) or variant of undetermined significance (n=21). Of the 104 patients who met 2010 revised Ghent criteria, 87/104 (82%) had a pathogenic or likely pathogenic variant. There was a significantly lower frequency of missense variants (41 vs 89%; P<0.0001) observed in the Ghent-positive (vs Ghent-negative) patients, and this association held true in age-based groupings. Previously described genotype associations with ectopia lentis and early onset/'neonatal' MFS were confirmed in our cohort. Overall, our study points to the imperfect nature of relying solely on clinical criteria to diagnose MFS as well as the potential importance of truncating/splicing variants in Ghent-positive cases. Furthermore, the description of numerous novel variants and associated clinical findings may be useful for future clinical interpretation of FBN1 genotype in patients with suspected MFS.


Mental-behavioral health data: 2001 NHIS.
These data highlights are based on analysis of the 2001 National Health Interview Survey (NHIS) public use data (http://www.cdc. gov/nchs/nhis.htm). NHIS is a multi-purpose survey conducted by the National Center for Health Statistics, Centers for Disease Control and Prevention. NHIS has been conducted continuously since 1957.


Cigarette smoking is associated with adverse pathological response and increased disease recurrence amongst patients with muscle-invasive bladder cancer treated with cisplatin-based neoadjuvant chemotherapy and radical cystectomy: a single-centre experience.
To investigate the association between smoking status and pathological response to cisplatin-based neoadjuvant chemotherapy (NAC) and survival outcomes in patients with muscle-invasive bladder cancer (MIBC) treated with radical cystectomy (RC). We reviewed 201 patients treated with NAC and RC for cT2-cT4N0M0 BC between 01/1999 and 01/2015. Smoking status was categorised as: 'never', 'former', and 'current' smoker. Pathological response to NAC was defined as: complete (ypT0N0), partial (ypTis/Ta/T1, N0), and no response (ypT2-4 or ypN+). Clinicopathological characteristics were analysed according to smoking status. Logistic regression analyses tested the association between smoking status and pathological response to NAC. Cox regression analyses tested risk factors associated with recurrence, overall (OM) and cancer-specific mortality (CSM). Overall, there were 58 (28.9%) never smokers, 87 (43.3%) former smokers, and 56 (27.9%) current smokers. No response to NAC was more frequently noted in current smokers (73.2%; P = 0.007). Former smoker (odds ratio [OR] 2.28; P = 0.024) and current smoker statuses (OR 4.52; P < 0.001) were significantly associated with no response to NAC, after adjusting for age, gender, Charlson Comorbidity Index, and clinical stage. Similarly, current smoking status (hazard ratio [HR] 2.14; P = 0.03) and extravesical pathological tumour stage (HR 3.31; P < 0.001) were independently associated with an increased risk of recurrence after RC. Cigarette smoking was significantly associated with adverse pathological response to cisplatin-based NAC in patients with MIBC treated with RC. Current smokers were at significantly higher risk of disease recurrence as compared to former and never smokers.


Noninvasive, repetitive, quantitative measurement of gene expression from a bicistronic message by positron emission tomography, following gene transfer with adenovirus.
Gene therapy protocols are hampered by the inability to monitor the location, magnitude, and duration of ectopic gene expression following DNA delivery. Consequently, it is difficult to establish quantitative correlations and/or causal relationships between therapeutic gene expression and phenotypic responses in treated individuals. One approach to monitor "therapeutic gene" expression indirectly is to incorporate reporter genes that can be imaged in vivo into bicistronic transcription units, along with the therapeutic genes. Expression of the dopamine D2 receptor (D2R) and herpes simplex virus thymidine kinase (HSV1-TK) can both be monitored, in vivo, by positron-emission tomography (PET). We created ad.DTm, an adenovirus containing a cytomegalovirus (CMV) early promoter-driven transcription unit, in which the D2R gene is placed proximal to an encephalomyocarditis virus internal ribosomal entry site (IRES) and a modified HSV1-tk gene is placed distal to the IRES. Following intravenous ad.DTm injection into mice, correlated hepatic D2R and HSV1-sr39tk PET reporter gene expression was demonstrated. Repeated microPET scanning quantitated both D2R-dependent sequestration of a positron-emitting ligand and HSV1-TK-dependent sequestration of a positron-emitting product. It is possible, in living mice, to investigate noninvasively and to measure quantitatively and repeatedly correlated expression of two coding regions from a bicistronic transcription unit over a 3-month period following adenovirus delivery.


Towards better models and mechanistic biomarkers for drug-induced gastrointestinal injury.
Adverse drug reactions affecting the gastrointestinal (GI) tract are a serious burden on patients, healthcare providers and the pharmaceutical industry. GI toxicity encompasses a range of pathologies in different parts of the GI tract. However, to date no specific mechanistic diagnostic/prognostic biomarkers or translatable pre-clinical models of GI toxicity exist. This review will cover the current knowledge of GI ADRs, existing biomarkers and models with potential application for toxicity screening/monitoring. We focus on the current gaps in our knowledge, the potential opportunities and recommend that a systematic approach is needed to identify mechanism-based GI biomarkers with potential for clinical translation.


Inferior Vena Cava Filters to Prevent Pulmonary Embolism: Systematic Review and Meta-Analysis.
Inferior vena cava (IVC) filters are widely used for prevention of pulmonary embolism (PE). However, uncertainty persists about their efficacy and safety. The authors conducted a systematic review and meta-analysis of the published reports on the efficacy and safety of IVC filters. The authors searched PubMed, the Cochrane Central Register of Controlled Trials, and ClinicalTrials.gov through October 3, 2016, for randomized controlled trials (RCTs) or prospective controlled observational studies of IVC filters versus none in patients at risk of PE. Inverse variance fixed-effects models with odds ratio (OR) as the effect measure were used for primary analyses. Main outcomes included subsequent PE, PE-related mortality, all-cause mortality, and subsequent deep vein thrombosis (DVT). The authors' search retrieved 1,986 studies, of which 11 met criteria for inclusion (6 RCTs and 5 prospective observational studies). Quality of evidence for RCTs was low to moderate. Overall, patients receiving IVC filters had lower risk for subsequent PE (OR: 0.50; 95% confidence interval [CI]: 0.33 to 0.75); increased risk for DVT (OR: 1.70; 95% CI: 1.17 to 2.48); nonsignificantly lower PE-related mortality (OR: 0.51; 95% CI: 0.25 to 1.05); and no change in all-cause mortality (OR: 0.91; 95% CI: 0.70 to 1.19). Limiting the results to RCTs showed similar results. Findings were substantively similar across a wide range of sensitivity analyses. Very few prospective controlled studies, with limited quality of evidence, exist regarding the efficacy and safety of IVC filters. Overall, filters appear to reduce the risk of subsequent PE, increase the risk for DVT, and have no significant effect on overall mortality.


Disease-modifying genes and monogenic disorders: experience in cystic fibrosis.
The mechanisms responsible for the determination of phenotypes are still not well understood; however, it has become apparent that modifier genes must play a considerable role in the phenotypic heterogeneity of Mendelian disorders. Significant advances in genetic technologies and molecular medicine allow huge amounts of information to be generated from individual samples within a reasonable time frame. This review focuses on the role of modifier genes using the example of cystic fibrosis, the most common lethal autosomal recessive disorder in the white population, and discusses the advantages and limitations of candidate gene approaches versus genome-wide association studies. Moreover, the implications of modifier gene research for other monogenic disorders, as well as its significance for diagnostic, prognostic, and therapeutic approaches are summarized. Increasing insight into modifying mechanisms opens up new perspectives, dispelling the idea of genetic disorders being caused by one single gene.


Aggregation Effects and Population-Based Dynamics as a Source of Therapy Resistance in Cancer.
Evolution of resistance allows cancer cells to adapt and continue proliferating even when therapy is initially very effective. Most investigations of treatment resistance focus on the adaptive phenotypic properties of individual cells. We propose that the resistance of a single cell to therapy may extend beyond its own phenotypic and molecular properties and be influenced by the phenotypic properties of surrounding cells and variations in cell density. Similar variation exists in population densities of animals living in groups and can significantly affect the outcome of an external threat. We investigate aggregation effects in cancer therapy using Darwinian models that integrate phenotypic properties of individual cells and common population effects found in nature to simulate the dynamics of resistance and sensitivity in the diverse cellular environments within cancers. We demonstrate that the density of cancer cell populations can profoundly influence response to chemotherapy independent of the properties of individual cells. Most commonly, these aggregation effects benefit the tumor allowing cells to survive even with phenotypic properties that would render them highly vulnerable to therapy in the absence of population effects. We demonstrate aggregation effects likely play a significant role in conferring resistance to therapy on tumor cells that would otherwise be sensitive to treatment. The potential role of aggregation in outcomes from cancer therapy has not been previously investigated. Our results demonstrate these dynamics may play a key role in resistance to therapy and could be used to design evolutionarily-enlightened therapies that exploit aggregation effects to improve treatment outcomes.


Electrophysiological population dynamics reveal context dependencies during decision making in human frontal cortex.
Evidence from monkeys and humans suggests that the orbitofrontal cortex (OFC) encodes the subjective value of options under consideration during choice. Data from non-human primates suggests that these value signals are context-dependent, representing subjective value in a way influenced by the decision makers' recent experience. Using electrodes distributed throughout cortical and subcortical structures, human epilepsy patients performed an auction task where they repeatedly reported the subjective values they placed on snack food items. High-gamma activity in many cortical and subcortical sites including the OFC positively correlated with subjective value. Other OFC sites showed signals contextually modulated by the subjective value of previously offered goods-a context dependency predicted by theory but not previously observed in humans. These results suggest that value and value-context signals are simultaneously present but separately represented in human frontal cortical activity.


Aging-related magnification of genetic effects on cognitive and brain integrity.
Heritability studies document substantial genetic influences on cognitive performance and decline in old age. Increasing evidence shows that effects of genetic variations on cognition, brain structure, and brain function become stronger as people age. Disproportionate impairments are typically observed for older individuals carrying disadvantageous genotypes of different candidate genes. These data support the resource-modulation hypothesis, which states that genetic effects are magnified in persons with constrained neural resources, such as older adults. However, given that findings are not unequivocal, we discuss the need to address several factors that may resolve inconsistencies in the extant literature (gene-gene and gene-environment interactions, study populations, gene-environment correlations, and epigenetic mechanisms).


An extract of Lycium barbarum mimics exercise to improve muscle endurance through increasing type IIa oxidative muscle fibers by activating ERRγ.
Lycium barbarum berry (gouqi, Goji, goji berry, or wolfberry), a traditional medicine and functional food, has a wide range of biological effects, including immuno-modulation, anti-aging, antitumor, neuro-protection, and hepato-protection. However, thus far, little is known about the traditional effects of L. barbarum on strengthening muscles. Therefore, this study focused on the effects of an extract of L. barbarum on skeletal muscles. First, the extract of L. barbarum significantly increased the mass of the tibial anterior muscle and gastrocnemius muscle and improved the average running distance of mice. Then, in vivo and in vitro experiments showed that the extract enhanced muscle endurance by increasing the proportion of type IIa oxidative muscle fibers and aerobic respiration. In an in-depth study of the molecular mechanism of these effects, we found that the extract upregulated the proportion of type IIa oxidative muscle fibers by activating ERRγ and that the PKA-CREB signaling pathway was involved in its activation. This study is the first to show that L. barbarum extract modulates skeletal muscle remodeling and has mimetic effects on skeletal muscles in a manner similar to exercise. It provides a scientific explanation based on modern biological technologies and concepts for the traditional function of L. barbarum in improving muscle fitness. This study lays a theoretical foundation for the application of L. barbarum in skeletal muscles as an exercise mimetic.


Prevalence of symptomatic urinary calculi in Kerman, Iran.
In a cross-sectional study, we evaluated the prevalence of symptomatic urinary calculi in Kerman, Iran. A total of 2431 citizens of Kerman were surveyed from September of 2005 to April 2006. The study population was selected by cluster method from 5 different areas, and 100 houses in each area were randomly approached. Questions on the urinary symptoms consisted of flank pain, urogenital pain, dysuria or any difficulty in urination, and alterations in urine color. Individuals with a suspicion of urinary calculi based on their symptoms were evaluated by physical examination, laboratory investigations, and plain abdominal radiography. Ultrasonography and intravenous urography were done if required to confirm the diagnoses. Of 2431 individuals, 196 (8.1%) had symptoms in favor of urinary calculus diagnosis, of whom 47 (24.0%) had urinary calculi (prevalence of symptomatic urolithiasis was 1.9%). Of the patients, 35 (74.5%) were underweight. Compared to the other participants, the patients with urinary calculus were younger (P = .001) and a larger proportion of them had a positive family history of urinary calculi (14.9% versus 6.5%; P = .02) and were rug weavers and office employees. Dependency on opium and its derivative was significantly more frequent in patients with urinary calculi (25.5% versus 0.2%; P = .001). This study showed that the prevalence of symptomatic urinary calculi in this hot and dry area is relatively high. According to our findings, the other factors including specific occupations, malnutrition, and substance use may also have influence on the rate of urinary calculus formation. Therefore, to prevention and early treatment of urinary calculi, evaluation of potential predisposing conditions should be considered with special attention to regional factors.


Protein engineering strategies for improving the selective methylation of target CpG sites by a dCas9-directed cytosine methyltransferase in bacteria.
Mammalian gene expression is a complex process regulated in part by CpG methylation. The ability to target methylation for de novo gene regulation could have therapeutic and research applications. We have previously developed a dCas9-MC/MN protein for targeting CpG methylation. dCas9-MC/MN is composed of an artificially split M.SssI methyltransferase (MC/MN), with the MC fragment fused to a nuclease-null CRISPR/Cas9 (dCas9). Guide RNAs directed dCas9-MC/MN to methylate target sites in E. coli and human cells but also caused some low-level off-target methylation. Here, in E. coli, we show that shortening the dCas9-MC linker increases methylation of CpG sites located at select distances from the dCas9 binding site. Although a shortened linker decreased methylation of other CpGs proximal to the target site, it did not reduce off-target methylation of more distant CpG sites. Instead, targeted mutagenesis of the methyltransferase's DNA binding domain, designed to reduce DNA affinity, significantly and preferentially reduced methylation of such sites.


A decrease in GTP content is associated with aerial mycelium formation in Streptomyces MA406-A-1.
Aerial mycelium formation by Streptomyces sp. MA406-A-1, a formycin-producing strain, was suppressed by the presence of excess nutrient. In such suppressed cultures, decoyinine, which specifically inhibits GMP synthetase, initiated the formation of aerial mycelium at concentrations which only partially inhibited growth. The intracellular GTP pool of organisms growing in liquid culture markedly decreased on the addition of decoyinine. Decoyinine was also effective in initiating aerial mycelium formation of three other Streptomyces spp. examined. Regardless of the successful initiation of aerial mycelium formation, the ability of the cells to produce antibiotics (formycin or actinomycin D) did not increase, but decreased, on the addition of decoyinine. It is concluded that aerial mycelium formation by Streptomyces results from a decrease in the pool of GTP (or GDP), whereas antibiotic synthesis results from a different signal(s).


Distribution of [3H] alpha, beta-methylene ATP binding sites in pulmonary blood vessels of different species.
Purinergic neurotransmission has been reported to take part in the regulation of pulmonary vascular resistance in many species. The receptor which mediates the contraction of blood vessels by ATP was defined as P2X-purinoceptors in pharmacological studies. In the present study, autoradiographic localization of P2X-purinoceptors was carried out in pulmonary blood vessels from rat, guinea-pig, rabbit, piglet and human by using [3H] alpha, beta-methylene ATP ([3H] alpha, beta-MeATP) as the radioligand. Autoradiographs showed that all the pulmonary arteries had been labelled with [3H] alpha, beta-MeATP. The specific binding sites were only associated with the smooth muscle of the blood vessels. Semi-quantitation of the autoradiographs revealed significant differences in the densities of P2X-purinoceptors amongst the vessels studied, both regional and interspecific. Generally, intrapulmonary arteries were labelled more heavily than the main pulmonary arteries, and the medium- and small-sized arteries had higher densities of P2X-purinoceptor than the large muscular arteries. The veins were sparsely labelled, except that rat intrapulmonary veins were labelled moderately. The results from this study provide: (1) direct evidence for the existence of P2X-purinoceptors in pulmonary vasculature, (2) a semi-quantitative estimation of the relative density of P2X-purinoceptors in pulmonary vessels of different sizes; and (3) a comparison of the P2X-purinoceptor densities among the species studied. Furthermore, the distribution of the P2X-purinoceptors is consistent with known pharmacological responses elicited by ATP in these vessels.


RNA interference-mediated silencing of the Hsp70 gene inhibits human gastric cancer cell growth and induces apoptosis in vitro and in vivo.
The role of heat shock protein (HSP) 70 in gastric cancer has been extensively examined in many studies for the past decade. It has been demonstrated that over-expression of Hsp70 might play important role in malignant transformation and maintenance of malignant phenotypes. Therefore, silencing the Hsp70 gene could be applicable in molecular therapies of human gastric cancer. Herein, we designed a small interfering RNA targeting Hsp70 to knock down its expression and investigated its effect on cell proliferation and apoptosis in a gastric cancer cell line. Two plasmids (phsp1-siRNA, phsp2-siRNA), along with a negative control (phsp3-siRNA), were created using a genic recombination technique. BGC823 cell lines were used to perform experiments. Western blotting and RT-PCR were used to detect Hsp70 expression in vitro and in vivo. Cell morphology was observed under light microscope. Cell cycle and apoptosis were analyzed by flow cytometry and acridine orange/ethidium bromide double stain, and cell proliferative activity was measured by alamarblue assay. In all experiments, a negative control served as a baseline measure. We successfully constructed phsps-siRNA plasmids and transfected them into BGC823 cells. RT-PCR and western blotting revealed that the expression of Hsp70 was down-regulated in transfection groups compared with the control group. Flow cytometric analysis indicated that less S-phase fraction accumulated in small interfering RNA transfected cells than in parental cells and the cells transfected with empty vector. Our results demonstrated that RNAi against Hsp70 could effectively knock down gene expression, inhibit growth of cancer cells, induce cell cycle arrest and increase cell apoptosis in vitro and in vivo. Hsp70 might serve as a therapeutic target for human gastric cancer.


Comparison of midazolam and propofol for sedation in pediatric diagnostic imaging studies.
This study aims to compare the efficacy of propofol and midazolam in terms of adverse effect potentials and to determine the appropriate strategy for pediatric procedural sedation. A total of 200 pediatric patients (aged < 14 years) undergoing diagnostic procedures were recruited for this nonrandomized prospective controlled cohort study. The patients were assigned to 2 treatment arms: either propofol (Group 1: IV bolus dose of 2 mg/kg during a 2-minute period, IV maintenance dose of 100 mcg/kg/min) or midazolam (Group 2: IV bolus dose of 0.15 mg/kg during a period of 2 to 3 minutes) to achieve sedation. Demographic data, body weight, and clinical status of the patients were evaluated and recorded. The vital signs and sedation levels (ie, Ramsay sedation scale scores) were evaluated and recorded, as well as the complications detected and medications administered in 10-minute intervals throughout the sedation procedure. Findings between the study arms were compared. Arterial blood pressures decreased significantly in both groups (P = 0.001). The patients in Group 1 experienced a greater difference in diastolic blood pressure (P = 0.001) than those in Group 2. Sedation scores in Group 1 were more favorable (P = 0.014) and reached the appropriate sedation level in a shorter time than those in Group 2 (P = 0.010). Likewise, recovery time of patients was shorter in Group 1 than in Group 2 (P = 0.010). Hypoxia was found to be more common in the propofol group, but the difference was not significant (P = 0.333). Propofol seems to be more effective, achieve the appropriate sedation level more quickly, and provide a faster onset of sedation than midazolam in pediatric procedural sedation and analgesia. Propofol is preferred for imaging studies (computed tomography and magnetic resonance imaging) to reduce the occurrence of undesired motion artefacts. Although both drugs are safe to use for sedation before pediatric imaging procedures, propofol is preferred with appropriate preparation.


COVID-19: pathophysiology, diagnosis, complications and investigational therapeutics.
The novel coronavirus disease 2019 (COVID-19) outbreak started in early December 2019 in the capital city of Wuhan, Hubei province, People's Republic of China, and caused a global pandemic. The number of patients confirmed to have this disease has exceeded 9 million in more than 215 countries, and more than 480 600 have died as of 25 June 2020. Coronaviruses were identified in the 1960s and have recently been identified as the cause of a Middle East respiratory syndrome (MERS-CoV) outbreak in 2012 and a severe acute respiratory syndrome (SARS) outbreak in 2003. The current SARS coronavirus 2 (SARS-CoV-2) is the most recently identified. Patients with COVID-19 may be asymptomatic. Typical symptoms include fever, dry cough and shortness of breath. Gastrointestinal symptoms such as nausea, vomiting, abdominal pain and diarrhoea have been reported; neurologically related symptoms, particularly anosmia, hyposmia and dysgeusia, have also been reported. Physical examination may find fever in over 44% of patients (and could be documented in over 88% of patients after admission), increased respiratory rate, acute respiratory disease and maybe decreased consciousness, agitation and confusion. This article aims at presenting an up-to-date review on the pathogenesis, diagnosis and complications of COVID-19 infection. Currently no therapeutics have been found to be effective. Investigational therapeutics are briefly discussed.


[A patient classification system for our primary care: the ambulatory care groups (ACGs)].
The ambulatory care groups (ACGs) were developed at the Johns Hopkins University in 1987 and they differ from other systems because their longitudinal approach and its capability to describe the case-mix of a population of reference. This paper describes the ACGs and assess its potential application to spanish primary care (PC). Observational study with the voluntary participation of 13 physicians (general practitioner and paediatrician) and nursing staff. During six months all diagnoses and interventions to a sample of attended patients were gathered. The first stage in grouping involves ascribing one of the 34 ambulatory diagnostic groups (ADGs) to each of the health problems attended. The second stage involves, after several intermediates steps, assigning one of the 51 ACGs to each patient. The total number of episodes was 7,559 (input), the number of patients (output) 2,467, the ICD codes with error 292, and the number of patients with ICD codes with error 278. With 5 ACGs we classify 44% of all the patients and with 10 ACGs, 64% of all them. Only two ACGs were empty. The article explores the potential applications of the system in spanish PC: for provider financing when a capitation payment component were introduced, utilization review and quality assurance when we compare several providers or populations, and research interested to link the process and its outcomes. Among the various patients classification system for grouping ambulatory care, the ambulatory care groups (ACGs) appear as the method with more potentialities to PC in Spain. The ACGs should be validated before been applied and its adaption to a more suitable PC classification of diagnoses would assists its extension.


Estrogen receptors in human bladder cells regulate innate cytokine responses to differentially modulate uropathogenic E. coli colonization.
The bladder epithelial cells elicit robust innate immune responses against urinary tract infections (UTIs) for preventing the bacterial colonization. Physiological fluctuations in circulating estrogen levels in women increase the susceptibility to UTI pathogenesis, often resulting in adverse health outcomes. Dr adhesin bearing Escherichia coli (Dr E. coli) cause recurrent UTIs in menopausal women and acute pyelonephritis in pregnant women. Dr E. coli bind to epithelial cells via host innate immune receptor CD55, under hormonal influence. The role of estrogens or estrogen receptors (ERs) in regulating the innate immune responses in the bladder are poorly understood. In the current study, we investigated the role of ERα, ERβ and GPR30 in modulating the innate immune responses against Dr E. coli induced UTI using human bladder epithelial carcinoma 5637 cells (HBEC). Both ERα and ERβ agonist treatment in bladder cells induced a protection against Dr E. coli invasion via upregulation of TNFα and downregulation of CD55 and IL10, and these effects were reversed by action of ERα and ERβ antagoinsts. In contrast, the agonist-mediated activation of GPR30 led to an increased bacterial colonization due to suppression of innate immune factors in the bladder cells, and these effects were reversed by the antagonist-mediated suppression of GPR30. Further, siRNA-mediated ERα knockdown in the bladder cells reversed the protection against bacterial invasion observed in the ERα positive bladder cells, by modulating the gene expression of TNFα, CD55 and IL10, thus confirming the protective role of ERα. We demonstrate for the first time a protective role of nuclear ERs, ERα and ERβ but not of membrane ER, GPR30 against Dr E. coli invasion in HBEC 5637 cells. These findings have many clinical implications and suggest that ERs may serve as potential drug targets towards developing novel therapeutics for regulating local innate immunity and treating UTIs.


Bacteriophages as an alternative for biological control of biofilm-forming Salmonella enterica.
Salmonellosis is one of the most common foodborne diseases worldwide. Surface adherence and biofilm formation are among the main strategies evolved by Salmonella to survive under harsh conditions and are risk factors for its spread through the food chain. Owing to the increase in antimicrobial resistance, there is a growing need to develop other methods to control foodborne pathogens, and bacteriophages have been suggested as a potential alternative for this purpose. The aim of this study was to evaluate bacteriophages as a biological control of Salmonella enterica serotypes to inhibit and remove bacterial biofilms. A total of 12 S. enterica isolates were selected for this study, all of which were biofilm producers. Seven bacteriophages were tested, individually and in a cocktail, for their host range and efficiency of plating (EOP). The phage cocktail was evaluated for its antibiofilm effect against the Salmonella biofilms. Phages UPF_BP1, UPF_BP2, UPF_BP3, UPF_BP6, and 10:2 possessed a broad lytic spectrum and could infect all S. enterica strains. Phages 10:2, UPF_BP6, and UPF_BP3 had high EOP in 10, 9, and 9 out of the 12 S. enterica strains, respectively. The cocktail was able to infect all S. enterica strains and had a high EOP in 10 out of 12 S. enterica isolates, presenting a broader host range than any of the tested single phages. A wide variation of inhibition among strains was observed, ranging from 14.72% to 88.53%. Multidrug-resistant and strong biofilm producer strains showed high biofilm inhibition levels by phage cocktail. Our findings demonstrate the ability of the cocktail to prevent biofilm formation and remove formed biofilms of Salmonella. These results indicate that the phage cocktail is a promising candidate to be used as an alternative for the control of Salmonella biofilms through surface conditioning.


[Effect of late motility on the fertilization of ova in the IVF program].
In the course of the IVF programme at our department we made the observation that even with primarily normal sperm counts late motility often varies widely. The impression that late sperm motility might be related to successful fertilisation caused us to study the influence of late sperm motility on fertilisation rates in a series of 80 patients. Sperm was capacitated with B2 medium (Menezo INRA). Even in the presence of an originally normal sperm count, reduced late motility was associated with lower fertilisation and embryo transfer rates. It seems that late sperm motility will have to be considered as an important parameter in evaluating the prognosis for successful IVF and ET.


Mouse hepatitis virus infection of the central nervous system: chemokine-mediated regulation of host defense and disease.
Infection of the central nervous system (CNS) of susceptible mice with mouse hepatitis virus (MHV), a positive-strand RNA virus that is a member of the Coronaviridae family, reproducibly results in an acute encephalomyelitis followed by a demyelinating disease similar to the human demyelinating disease multiple sclerosis (MS). MHV infection triggers a robust cell-mediated response in which both CD4+ and CD8+ T cells are essential in controlling viral replication and spread. However, viral clearance is incomplete and viral RNA and protein can persist within white matter tracts, areas of viral persistence are often associated with demyelinating lesions, and recent studies have indicated an important role for both T cells and macrophages in contributing to myelin destruction. The molecular mechanisms governing leukocyte trafficking and accumulation within the CNS of MHV-infected mice are just now being understood and recent studies indicate that chemokines and chemokine receptors have an important role in this process. This article will provide an overview on how these molecules regulate T cell and macrophage trafficking into the CNS of MHV-infected mice and illustrate the delicate balance that exists with regards to expression of chemokines and their receptors as it relates to both host defense and disease development.


The national burden of poorly controlled asthma, school absence and parental work loss among school-aged children in the United States.
The degree of poorly controlled asthma and its association with missed school days and parental missed work days is not well understood. This was a retrospective analysis of missed school days and missed work days for school-aged children (SAC; aged 6-17) and their caregivers in the nationally representative 2007-2013 Medical Expenditure Panel Survey (MEPS). Indicators of poor asthma control included: exacerbation in previous 12 months; use of >3 canisters of short-acting beta agonist (SABA) in 3 months; and annual asthma-specific (AS) Emergency Department (ED) or inpatient (IP) visits. Negative binomial regression was used for missed school days, and a Heckman two-step selection model was used for missed work days. All analyses controlled for sociodemographics and other covariates. There were 44,320 SAC in MEPS, of whom 5,890 had asthma. SAC with asthma and an indicator of poor control missed more school days than SAC without asthma: exacerbation (1.8 times more; p < 0.001); >3 canisters SABA (2.7 times more; p < 0.001) and ED/IP visit (3.8 times more; p < 0.001). The parents/caregivers of SAC with asthma and an exacerbation missed 1.2 times more work days (p < 0.05), while those with SAC with asthma and an ED/IP visit missed 1.8 times more work days (p < 0.01) than the parents of SAC without asthma. This study provides evidence of the significant national burden of poorly controlled asthma due to missed school and work days in the United States. More effective and creative asthma management strategies, with collaboration across clinical, community and school-based outreach, may help address this burden.


Morphine (10, 20 mg) in a Postoperative Dressing Used with Patients After Surgical Debridement of Burn Wounds: A Prospective, Double-Blinded, Randomized Controlled Trial.
Objective: This is the first clinical trial to evaluate the analgesic effect of 10 and 20 mg of morphine used in a postoperative dressing with patients after surgical debridement of burn wounds. Approach: In this randomized controlled trial, 20 adult patients with third-degree flame burns, who had undergone surgical debridement under general anesthesia, were randomly assigned to either group A, whose members were treated with a burn dressing that contained 10 mg of morphine, or group B, whose members were treated with a burn dressing that contained 20 mg of morphine; the dressing was also soaked with octenidine and phenoxyethanol in the case of both groups. The plasma morphine concentrations were measured 1, 2, 3, and 6 h after surgery, while the level of pain intensity was determined on the Numeric Pain Rating Scale (NRS), and the occurrence of side effects was observed. Results: The serum morphine concentration levels were very low, but statistically different between the two groups at all time points. The NRS value was similar in both groups at all time points (p > 0.05). Despite this, in group B, the NRS value was 0 in all patients in postoperative hours 1, 2, and 3. No adverse effect of morphine sulfate was observed in any patient. Innovation: This project is the first clinical study to have demonstrated that morphine administered in dressings in concentrations of 0.02-0.08 mg/mL significantly reduces the occurrence of pain. Conclusion: The use of morphine in dressings after surgical treatment of burn wounds is very effective when it comes to pain management and is safe for the patient.


Electrostatic spray ionization mass spectrometry imaging.
Imaging samples on a surface by mass spectrometry (MS) requires the combination of MS detection with a scanning mode that enables localized desorption and ionization and/or detection of sample analytes with good spatial resolution. We have developed a new mass spectrometry imaging (MSI) method based on electrostatic spray ionization. It works under ambient conditions and can be applied to a wide range of molecules providing quantitative MS analysis even in the presence of salts in excess. 2D MS images of protein and peptide spots, inkjet-printed black dye patterns, and cells were obtained. The presented novel ambient ionization mass spectrometry imaging method can find many applications in analytical and bioanalytical chemistry.


Effects of Adsorbed Polyaniline on Redox Process on MoO3 Surface.
The effects exhibited by adsorbed conducting polyaniline on the redox process on a molybdenum oxide surface were studied. Thermogravimetric results indicate a 4% polyaniline deposition. Cyclic voltammograms of the adsorbed polymer on MoO3 show that polyaniline exerts remarkable effects on the molybdenum blue oxidation-reduction process, with oxidation and reduction potentials of 0.33 and 0.18 V, respectively. This effect strongly enhances the electrode response, and can be used as an important tool in qualitative and/or quantitative determinations of molybdenum in solution as well as in any substrate. Copyright 1999 Academic Press.


Multicenter study of high-dose daptomycin for treatment of enterococcal infections.
Enterococci are among the leading pathogens isolated in hospital-acquired infections. Current antimicrobial options for vancomycin-resistant enterococci (VRE) are limited. Prior data suggest that daptomycin at >6 mg/kg of body weight/day may be used to treat enterococcal infections. We retrospectively evaluated the effectiveness and safety of high-dose daptomycin (HD-daptomycin) therapy (>6 mg/kg) in a multicenter cohort of adult patients with enterococcal infections to describe the characteristics and outcomes. Two hundred forty-five patients were evaluated. Enterococcus faecium was identified in 175 (71%), followed by Enterococcus faecalis in 49 (20%) and Enterococcus spp. in 21 (9%); overall, 204 (83%) isolates were VRE. Enterococcal infections included bacteremia (173, 71%) and intra-abdominal (35, 14%) and bone and joint (25, 10%) infections. The median dosage and duration of HD-daptomycin were 8.2 mg/kg/day (interquartile range [IQR], 7.7 to 9.7) and 10 days (IQR, 6 to 15), respectively. The overall clinical success rate was 89% (193/218), and microbiological eradication was observed in 93% (177/191) of patients. The median time to clearance of blood cultures on HD-daptomycin was 3 days (IQR, 2 to 5). The 30-day all-cause mortality rate was 27%, and 5 (2%) patients developed daptomycin-nonsusceptible enterococcal strains while on HD-daptomycin. Seven patients (3%) had creatine phosphokinase (CPK) elevation, yet no HD-daptomycin regimen was discontinued due to an elevated CPK and all patients were asymptomatic. Overall, there was a high frequency of clinical success and microbiological eradication in patients treated with HD-daptomycin for enterococcal infections, even in patients with complicated and difficult-to-treat infections. No adverse event-related discontinuation of HD-daptomycin was noted. HD-daptomycin may be an option for the treatment of enterococcal infections.


Transition From Hospital to Home Following Hematopoietic Stem Cell Transplantation: A Feasibility Study for "Rooming in".
Background: The experience of hematopoietic stem cell transplant (HSCT) on both the patients and their caregivers is complex and challenging during hospitalization and post discharge. Complex patient populations require heightened attention on discharge practices to ensure that they are prepared for home regimens. "Rooming in" is a standardized intervention implemented prior to discharge that allows patients and caregivers to assume post discharge care with the support of staff. Other complex patient populations have reported positive outcomes related to "rooming in." Aims: The purpose of this study was to assess the feasibility of a standardized "rooming in" intervention for discharge of pediatric HSCT patients. An additional aim was to describe the quality of discharge teaching, readiness for hospital discharge, and postdischarge coping difficulty in a cohort of HSCT patients using validated questionnaires. Method: Data were collected via medical chart review. A prospective cohort completed validated study questionnaires at discharge and 30 days postdischarge. Results: All caregivers of post-HSCT patients were able to complete the "rooming in" intervention. There was no statistically significant difference for length of stay between the retrospective and prospective cohorts. Caregivers enrolled on the study rated the Quality of Discharge Teaching Scale-Parent Form high (Mdn = 165). Conclusion: We conclude that "rooming in" is a feasible discharge intervention for caregivers of pediatric HSCT patients.


An overview of the neuro-cognitive processes involved in the encoding, consolidation, and retrieval of true and false memories.
Perception and memory are imperfect reconstructions of reality. These reconstructions are prone to be influenced by several factors, which may result in false memories. A false memory is the recollection of an event, or details of an episode, that did not actually occur. Memory formation comprises at least three different sub-processes: encoding, consolidation and the retrieval of the learned material. All of these sub-processes are vulnerable for specific errors and consequently may result in false memories. Whereas, processes like imagery, self-referential encoding or spreading activation can lead to the formation of false memories at encoding, semantic generalization during sleep and updating processes due to misleading post event information, in particular, are relevant at the consolidation stage. Finally at the retrieval stage, monitoring processes, which are assumed to be essential to reject false memories, are of specific importance. Different neuro-cognitive processes have been linked to the formation of true and false memories. Most consistently the medial temporal lobe and the medial and lateral prefrontal cortex have been reported with regard to the formation of true and false memories. Despite the fact that all phases entailing memory formation, consolidation of stored information and retrieval processes, are relevant for the forming of false memories, most studies focused on either memory encoding or retrieval. Thus, future studies should try to integrate data from all phases to give a more comprehensive view on systematic memory distortions. An initial outline is developed within this review to connect the different memory stages and research strategies.


Influence of mercury ions on electrical properties of rat proximal and distal renal tubules.
The present study was designed to elucidate the acute effects of mercury ions on electrogenic transport systems in rat proximal and distal convoluted tubules. Male Munich Wistar rats were anesthetized with Inactin and prepared for micropuncture in the usual way. Under control conditions, the transepithelial potential difference (PDte) approaches -3.2 +/- 0.6 mV in early proximal tubules, +2.1 +/- 0.3 mV in late proximal tubules, and -17.8 +/- 1.8 mV in distal convoluted tubules. PDte of neither early nor late proximal tubules was significantly affected by the acute application of mercury ions. In the distal tubules, mercury ions (10 mumol/l) lead to a gradual depolarization of the epithelium from -17.8 +/- 1.8 to -9.3 +/- 2.7 mV within 8 min. A linear correlation between PDte under control conditions and depolarization caused by amiloride is seen. The slope of this correlation is significantly reduced during the application of mercury ions. In conclusion, mercury ions decrease the distal PDte an effect possibly contributing to altered renal electrolyte excretion following the acute administration of mercury ions. The depolarization is, at least in part, due to an inhibition of amiloride-sensitive sodium channels at the luminal cell membrane of principal cells.


Pediatric analgesia and sedation for the management of orthopedic conditions.
Orthopedic injuries are common in the emergency department; this is especially true in children. The orthopedist must often supervise sedation and analgesia for the injured child. While the techniques of conscious sedation have improved, the therapeutic index between adequate and excessive sedation is much narrower in children than adults, especially for painful procedures. The purpose of this review is to outline a reasonable approach to sedation and analgesia in the pediatric patient. In addition, current monitoring guidelines are reviewed, as well as the basic pharmacology of the most commonly used drugs.


Management of Primary Plasma Cell Leukemia Remains Challenging Even in the Era of Novel Agents.
Primary plasma cell leukemia (PCL) is a rare and aggressive variant of multiple myeloma (MM). PCL is characterized by peripheral blood involvement by malignant plasma cells and an aggressive clinical course leading to poor survival. There is considerable overlap between MM and PCL with respect to clinical, immunophenotypic, and cytogenetic features, but circulating plasma cell count exceeding 20% of peripheral blood leukocytes or an absolute plasma cell count of >2000/mm3 distinguishes it from MM. After initial stabilization and diagnosis confirmation, treatment of PCL in a fit patient typically includes induction combination chemotherapy containing novel agents typically, with proteasome inhibitors (such as bortezomib) and immunomodulatory drugs (eg, lenalidomide), followed by autologous hematopoietic stem cell transplant (HSCT) and multidrug maintenance therapy using novel agents post-HSCT. Long-term outcomes have improved employing this strategy but the prognosis for non-HSCT candidates remains poor and new approaches are needed for such PCL patients not eligible for HSCT. Here, we report a case of primary PCL, and a comprehensive and up to date review of the literature for diagnosis and management of PCL. We also present the findings of Positron Emission Tomography (PET) scan. Since PCL is often associated with extra-medulary disease, including PET scan at the time of staging and restaging may be a novel approach particularly to evaluate the extra-medullary disease sites.


Implantation of stents for treatment of recurrent and native coarctation in children weighing less than 20 kilograms.
We report our experience with implantation of stents for treatment of recurrent and native aortic coarctation in children weighing less than 20 kilograms. We treated 9 such patients between March, 2003, and January, 2006. In 2 patients, the coarctation had not previously been treated, while in 7 it had recurred after surgery. The patients had a median weight of 14 kilograms, with a range from 5.5 to 19 kilograms. Balloon dilation was needed in 1 patient before the stent was implanted. We used Palmaz Genesis XD stents in 7 patients, these having lengths from 19 to 29 millimetres, 1 Palmaz Genesis 124P stent, and 1 peripheral JoStent with a diameter of 6 to 12 millimetres. Implantation was effective in all patients. Immediately after implantation, the mean peak systolic gradient decreased from 30 millimetres of mercury, the range having been 15 to 50 mm, to 3 millimetres of mercury, with the final range from zero to 10 mm. There were no complications, with no observations of aneurysms, dissections, or dislocated stents. In 1 patient, the peripheral pulse was weak secondary to arterial access, but treatment with Heparin led to complete resolution. It was necessary to re-dilate the stent in another patient, while 2 others are scheduled for redilation because of growth-related restenosis. Our findings suggest that implantation of stents can produce excellent relief of the gradient produced by recurrent or native coarctation. The process is safe and effective in patients weighing less than 20 kilograms.


Otic barotrauma from air travel.
Otic barotrauma occurring during air travel involves traumatic inflammation of the middle ear, caused by a pressure difference between the air in the middle ear and the external atmosphere, developing after ascent or more usually descent. The pressure difference occurs because of failure of the eustachian tube to equilibrate middle ear and atmospheric pressures. It is a common problem, presenting with ear fullness, otalgia and deafness. Severe cases may result in tympanic membrane perforation and even round window membrane rupture. Of three randomized controlled trials, one showed that oral pseudoephedrine decongestants reduced otalgia in adults with recurrent ear pain during air travel, whilst another found that oral pseudoephedrine did not decrease in-flight ear pain in children. The third trial showed that oxymetazoline decongestant nasal spray, taken 30 minutes before descent, did not produce a statistically significant reduction in symptoms of barotrauma in adults with recurrent ear pain during air travel. We review the causes, prevention and treatment of this condition.


Shoulder Damage Model and Its Application for Single Point Diamond Machining of ZnSe Crystal.
The damaging of ZnSe crystal has a significant impact on its service performance and life. Based on the specific cutting energies for brittle and ductile mode machining, a model is proposed to evaluate the damage depth in the shoulder region of ZnSe crystal during single point diamond machining. The model considers the brittle-ductile transition and spring back of ZnSe crystal. To verify the model, the elastic modulus, hardness, spring back, and friction coefficient of ZnSe crystal are measured by nanoindentation and nanoscratch tests, and its critical undeformed chip thickness is obtained by spiral scratching. Meanwhile, orthogonal cutting experiments are conducted to obtain the different shoulder regions and cutting surfaces. The shoulder damage depth is analyzed, indicating that the effect of the feed on the damage depth at a high cutting depth is stronger than that at a low one. The model is verified to be effective with an average relative error of less than 7%. Then, the model is used to calculate the critical processing parameters and achieve a smooth ZnSe surface with a roughness Sa = 1.0 nm. The model is also extended to efficiently predict the bound of the subsurface damage depth of a cutting surface. The research would be useful for the evaluation of surface and subsurface damages during the ultra-precision machining of ZnSe crystal.


Recurrent autoimmune inner ear disease (AIED) and polyarteritis nodosa in a patient with large cell lung carcinoma.
Autoimmune inner ear disease (AIED) is a very rare disorder with distinct clinical features and can occur in patients with malignancy or autoimmune diseases. We report a 72-year-old male patient with polyarteritis nodosa treated continuously for 5 years with aggressive immunosuppressive drugs, including cyclophosphamide, who experienced three episodes of acute hearing loss during treatment. Organic lesions of the external and middle ear were excluded by repeated examinations, and if one subscribes to McCabe's (Ann Otol Rhinol Laryngol 88:585-589, 1979) definition of AIED, this condition must be considered as the likely cause of the hearing loss. During the period of treatment, three episodes of AIED occurred, and eventually, lung cancer developed. From the time relationship and clinical manifestations of neuropathy and livedo reticularis, the first episode of hearing loss was more likely to be related to vasculitis itself, while the third episode may well have been associated with the development of lung cancer given the dramatic improvement in the clinical condition following treatment of the tumor by excision and cancer chemotherapy. Coexistence of AIED, vasculitis, and malignancy in the same patient has only been reported infrequently, and our case suggests that this coexistence may not be coincidental. For those patients with autoimmune disease who are on long-term immunosuppressive drug therapy, active surveillance for a nascent malignant tumor should be exercised if AIED recurs or persists.


PIK3CA amplification is associated with poor prognosis among patients with curatively resected esophageal squamous cell carcinoma.
To investigate the clinicopathologic characteristics and the prognostic impact of PIK3CA gene amplification in curatively resected esophageal squamous cell carcinoma (ESCC). Using 534 curatively resected ESCCs, the PIK3CA gene copy number was evaluated with fluorescent in situ hybridization. PIK3CA amplification was defined as PIK3CA/centromere 3 ratio is ≥ 2.0 or average number of PIK3CA signals/tumor cell nucleus ≥ 5.0. PIK3CA mutations in exon 9 and 20, encoding the highly conserved helical and kinase domains were assessed by direct sequencing in 388 cases. PIK3CA amplification was detected in 56 (10.5%) cases. PIK3CA amplification was significantly associated with higher T-stage (P=0.026) and pathologic stage (P=0.053). PIK3CA amplification showed a significantly shorter disease free survival (DFS) compared with that of non-amplified group (33.4 vs 63.1 months, P=0.019). After adjusting for gender, tumor location, pathologic stage, histologic grade and adjuvant treatment, PIK3CA amplification was significantly associated with a shorter DFS (adjusted hazard ratio [AHR] 1.53; 95% CI, 1.10-2.17; P=0.02). Though the statistical insignificance, PIK3CA amplification showed tendency of shorter OS (52.1 vs 96.5 moths, P=0.116). PIK3CA mutations were detected in 6 (1.5%) of 388 cases; 5 cases with exon 9 mutations in E545K while one exon 20 mutation in H1047L. PIK3CA amplification is a frequent oncogenic alteration and associated with shorter survival, suggesting its role as a prognostic biomarker in resected ESCC. PIK3CA amplification may represent a promising therapeutic target for ESCC.


Spectral characteristics of air and bone conduction transducers used to record the auditory brain stem response.
This study sought to determine differences in the acoustic spectra of five different transducers commonly used for stimulus presentation to record the auditory brain stem response (ABR). The outputs of three commercially available bone conduction vibrators (Radioear B-70A, B-71 and B-72), a TDH-49 earphone, and an insert receiver were measured by applying a 0.1 msec rectangular electrical pulse to each transducer. The resultant output for each transducer was converted to one-third octave band data and plotted against reference threshold levels. Results demonstrated relatively flat acoustic spectra and high output levels for the two air conduction receivers. In contrast, each of the bone oscillators had its greatest concentration of energy in the 2000 Hz region with the spectrum characterized by a precipitous decrease in output at frequencies above and below this resonance peak. Maximum output never exceeded 35 dB HL for any of the three bone conduction devices. Of the three oscillators, however, the B-70A appeared to provide the highest output before reaching saturation. Results are discussed relative to the limitations for recording the auditory brain stem response to bone conducted transient signals.


Y-complex nucleoporins independently contribute to nuclear pore assembly and gene regulation in neuronal progenitors.
Besides assembling nuclear pore complexes, the conduits of nuclear transport, many nucleoporins also contribute to chromatin organization and gene expression, with critical roles in development and pathologies. We previously reported that Nup133 and Seh1, two components of the Y-complex subassembly of the nuclear pore scaffold, are dispensable for mouse embryonic stem cell viability but required for their survival during neuroectodermal differentiation. Here, a transcriptomic analysis revealed that Nup133 regulates a subset of genes at early stages of neuroectodermal differentiation, including Lhx1 and Nup210l, which encodes a newly validated nucleoporin. These genes are also misregulated in Nup133ΔMid neuronal progenitors, in which nuclear pore basket assembly is impaired. However, a four-fold reduction of Nup133 levels, despite also affecting basket assembly, is not sufficient to alter Nup210l and Lhx1 expression. Finally, these two genes are also misregulated in Seh1-deficient neural progenitors, which only show a mild reduction in nuclear pore density. Together these data reveal a shared function of Y-complex nucleoporins in gene regulation during neuroectodermal differentiation, apparently independent of nuclear pore basket integrity.


Pigment dispersion syndrome masquerading as acute anterior uveitis.
Signs and symptoms of pigment dispersion may be confused with those of acute anterior uveitis. This case series is intended to aid the ophthalmologist in the clinical differentiation between these two disorders. The authors present a series of 6 patients with pigment dispersion who were initially diagnosed as having acute anterior uveitis and treated with anti-inflammatory medication, including corticosteroids. The patients were referred for a second opinion due to poor or no response to therapy and were found to have pigment dispersion instead of uveitis. Symptoms of pigment dispersion may consist of blurred vision, redness, ocular pain, and photophobia, all of which are also symptoms of acute anterior uveitis. These symptoms, plus the fact that pigment floating in the aqueous humor can be mistaken for inflammation, make diagnosis challenging. Moreover, the possible co-existence of true anterior uveitis and pigment dispersion makes the diagnosis and treatment more difficult.


Contrast-Enhanced Ultrasound Assessment of Renal Parenchymal Perfusion in Patients with Atherosclerotic Renal Artery Stenosis to Predict Renal Function Improvement After Revascularization.
Identifying patients with atherosclerotic renal artery stenosis (ARAS) who will be improved in renal function after percutaneous transluminal renal artery stenting (PTRAS) is crucial since most patients show no worthwhile benefit of PTRAS. Although the assessment of renal parenchymal perfusion is useful for the identification, few studies predict the renal functional improvement by evaluating the characteristics of renal perfusion. The aim of this study was to assess the renal parenchymal perfusion in ARAS patients with contrast-enhanced ultrasonography (CEUS) and predict the benefits of renal function after PTRAS utilizing time-intensity curve (TIC) parameters. Thirty-eight kidneys in 30 ARAS patients received PTRAS in this study. They were divided into moderate stenosis group (n=25) and severe stenosis group (n=13) and mild dysfunction group (n=14) and moderate dysfunction group (n=24) according to the degree of renal stenosis and radioisotope glomerular filtration rate (rGFR). The baseline assessment of renal function and renal parenchymal perfusion were performed for all patients. rGFR was repeated to evaluate the renal outcome at 4 months after PTRAS. The outcome of PTRAS was classified as improved, stable, or deteriorated compared to the baseline. Time-intensity curve (TIC) parameters obtained from CEUS were analyzed to evaluate the predictive accuracy. TIC parameters (AUC and PI) were positively correlated with renal function (r=0.617, 0.663; P<0.05) but weakly and negatively correlated with the stenosis (r=-0.360, -0.435; P<0.05). Baseline rGFR was not accurate in predicting improved renal function after PTRAS (0.670). The accuracy of the combined prediction model of baseline AUC and PI (0.889) was higher than the individual indicators (baseline AUC: 0.855 and PI: 0.782). CEUS could accurately assess renal parenchymal perfusion and identify ARAS patients with potential benefit after PTRAS. The combination of TIC parameters (AUC and PI) is valuable in the prediction of improved renal function after PTRAS.


The importance of efficacy and partial agonism in evaluating models of B lymphocyte activation.
Immunologists have developed a range of in vitro techniques for probing the receptor mediated response of cells comprising the immune system. An important and ubiquitous method is the use of antibodies in either soluble or aggregated form to engage cell surface receptors and transmit a signal. Models of cell and molecular interactions, derived from the use of these antibodies, form the basis of our efforts to understand and explain the corresponding in vivo systems. However, interpreting in vitro experiments and distinguishing between alternative models is difficult. This complexity is illustrated here using B cell stimulation by surface immunoglobulin and CD40. The fluorescent cell labelling dye carboxyfluorescein, diacetate, succinimidyl ester (CFSE) is used to show that many anti-Ig and CD40 stimulatory agents, used to assess the role of B cells and lymphokines, are partial agonists. By modelling each step in B cell signalling, activation and division it is possible to show that small changes in signal contributed by a second receptor can generate numerous distinct dose response curves that are highly dependent on the "efficacy" of signal transmission by the primary ligand and the number of cell divisions taken in culture. Differences in dose response curves become particularly striking if the primary activating stimulus is a partial agonist. Although exemplified here with B cell stimulation the conclusions are applicable to other in vitro activation systems and suggest ways to improve both the design and interpretation of in vitro experiments.


An approach to multiobjective optimization of rotational therapy. II. Pareto optimal surfaces and linear combinations of modulated blocked arcs for a prostate geometry.
The purpose of this work is twofold: To further develop an approach to multiobjective optimization of rotational therapy treatments recently introduced by the authors [J. Pardo-Montero and J. D. Fenwick, "An approach to multiobjective optimization of rotational therapy," Med. Phys. 36, 3292-3303 (2009)], especially regarding its application to realistic geometries, and to study the quality (Pareto optimality) of plans obtained using such an approach by comparing them with Pareto optimal plans obtained through inverse planning. In the previous work of the authors, a methodology is proposed for constructing a large number of plans, with different compromises between the objectives involved, from a small number of geometrically based arcs, each arc prioritizing different objectives. Here, this method has been further developed and studied. Two different techniques for constructing these arcs are investigated, one based on image-reconstruction algorithms and the other based on more common gradient-descent algorithms. The difficulty of dealing with organs abutting the target, briefly reported in previous work of the authors, has been investigated using partial OAR unblocking. Optimality of the solutions has been investigated by comparison with a Pareto front obtained from inverse planning. A relative Euclidean distance has been used to measure the distance of these plans to the Pareto front, and dose volume histogram comparisons have been used to gauge the clinical impact of these distances. A prostate geometry has been used for the study. For geometries where a blocked OAR abuts the target, moderate OAR unblocking can substantially improve target dose distribution and minimize hot spots while not overly compromising dose sparing of the organ. Image-reconstruction type and gradient-descent blocked-arc computations generate similar results. The Pareto front for the prostate geometry, reconstructed using a large number of inverse plans, presents a hockey-stick shape comprising two regions: One where the dose to the target is close to prescription and trade-offs can be made between doses to the organs at risk and (small) changes in target dose, and one where very substantial rectal sparing is achieved at the cost of large target underdosage. Plans computed following the approach using a conformal arc and four blocked arcs generally lie close to the Pareto front, although distances of some plans from high gradient regions of the Pareto front can be greater. Only around 12% of plans lie a relative Euclidean distance of 0.15 or greater from the Pareto front. Using the alternative distance measure of Craft ["Calculating and controlling the error of discrete representations of Pareto surfaces in convex multi-criteria optimization," Phys. Medica (to be published)], around 2/5 of plans lie more than 0.05 from the front. Computation of blocked arcs is quite fast, the algorithms requiring 35%-80% of the running time per iteration needed for conventional inverse plan computation. The geometry-based arc approach to multicriteria optimization of rotational therapy allows solutions to be obtained that lie close to the Pareto front. Both the image-reconstruction type and gradient-descent algorithms produce similar modulated arcs, the latter one perhaps being preferred because it is more easily implementable in standard treatment planning systems. Moderate unblocking provides a good way of dealing with OARs which abut the PTV. Optimization of geometry-based arcs is faster than usual inverse optimization of treatment plans, making this approach more rapid than an inverse-based Pareto front reconstruction.


Thiazolidinediones enhance sodium-coupled bicarbonate absorption from renal proximal tubules via PPARγ-dependent nongenomic signaling.
Thiazolidinediones (TZDs) improve insulin resistance by activating a nuclear hormone receptor, peroxisome proliferator-activated receptor γ (PPARγ). However, the use of TZDs is associated with plasma volume expansion through a mechanism that remains to be clarified. Here we showed that TZDs rapidly stimulate sodium-coupled bicarbonate absorption from the renal proximal tubule in vitro and in vivo. TZD-induced transport stimulation is dependent on PPARγ-Src-EGFR-ERK and observed in rat, rabbit and human, but not in mouse proximal tubules where Src-EGFR is constitutively activated. The existence of PPARγ-Src-dependent nongenomic signaling, which requires the ligand-binding ability, but not the transcriptional activity of PPARγ, is confirmed in mouse embryonic fibroblast cells. The enhancement of the association between PPARγ and Src by TZDs supports an indispensable role of Src in this signaling. These results suggest that the PPARγ-dependent nongenomic stimulation of renal proximal transport is also involved in TZD-induced volume expansion.


In vitro fungistatic effects of natural coniferous resin from Norway spruce (Picea abies).
Resins (rosin, pitch) are natural products of the coniferous trees and are antimicrobial against a wide range of microbes. The antifungal effectiveness of resin, purified from Norway spruce (Picea abies), was studied against human pathogenic fungi and yeasts with the agar plate diffusion tests and electron microscopy (EM). The fungistatic effect of these resin mixtures (resin salves) was tested against a set of Candida yeasts, dermatophytes, and opportunistic fungi. Transmission and scanning EM was done from samples of fungi (Trichophyton mentagrophytes). In agar diffusion tests, the resin was strongly antifungal against all dermatophytes tested, e.g., against all fungi of the genus Trichophyton, but it was not antifungal against the Candida yeasts or against the opportunistic fungi tested. According to EM, resin caused damages in the cell hyphae and cell wall structures. We conclude that, in the agar plate diffusion test, coniferous resins are strongly fungistatic against the dermatophytic fungi only.


Regulation of IGF binding protein-1 in hep G2 cells by cytokines and reactive oxygen species.
The liver is a major site of synthesis for insulin-like growth factor binding protein (IGFBP)-1. Because IGFBP-1 inhibits many anabolic actions of IGF-I, increases in IGFBP-1 may be partly responsible for the decrease in lean body mass observed in catabolic/inflammatory conditions. This study aimed to determine in Hep G2 cells 1) the sensitivity of IGFBP-1 synthesis to treatment with interleukin (IL)-1, tumor necrosis factor-alpha (TNF-alpha), and IL-6, 2) the ability of reactive oxygen species (ROS) to enhance IGFBP-1 production, and 3) the role of ROS in mediating cytokine-induced increases in IGFBP-1. Hep G2 cells responded to IL-1beta, TNF-alpha, and IL-6 with maximal 8- to 10-fold increases in IGFBP-1 production. Although the maximal responsiveness of cells treated with TNF-alpha and IL-6 was 20-30% less than that with IL-1beta, cells demonstrated a similar sensitivity to all cytokines (half-maximal responsive dose of approximately 10 ng/ml). A low concentration (3 ng/ml) of all three cytokines had an additive effect on IGFBP-1 production. Cytokines also increased IGFBP-1 mRNA. The half-life of IGFBP-1 mRNA was approximately 4 h and not altered by IL-1beta. Incubation with ROS, including H2O2 and nitric oxide (NO) donors, resulted in a relatively smaller increase in IGFBP-1. However, preincubating Hep G2 cells with various free radical scavengers and NO synthase and eicosanoid inhibitors failed to prevent or attenuate cytokine-induced increases in IGFBP-1. Finally, preincubating cells with pyrrolidinedithiocarbamate (PDTC) but not SN50 (inhibitors of nuclear factor-kappaB activation and nuclear translocation, respectively) attenuated increases in IGFBP-1 induced by IL-1. These results indicate that 1) proinflammatory cytokines directly enhance IGFBP-1 synthesis by stimulating transcription without altering mRNA stability, 2) addition of exogenous ROS also stimulates IGFBP-1 production but to a smaller extent than cytokines, and 3) the cytokine-induced increase in IGFBP-1 production is not mediated by endogenous production of ROS or eicosanoids but appears to at least partially involve a PDTC-sensitive pathway.


Retinal artery emboli. Indications for angiography and carotid endarterectomy.
Six patients with ocular symptoms were referred by the Eye Service to the Vascular Service because of the presence of cholesterol emboli on fundoscopic examination of the retinal arteries. None of the six patients had classic intermittent retinal or cerebral ischemic attacks. Four-vessel aortic arch arteriogram was suggested and significant ipsilateral carotid disease was found in all patients. Four patients underwent carotid endarterectomy, with removal of ulcerated plaques from the carotid bifurcation. Two patients had total occlusion of the ipsilateral internal carotid artery and therefore were not operative candidates. The presence of retinal cholesterol emboli is an indication for extracranial arteriography. When ipsilateral ulcerative disease is found, carotid endarterectomy is indicated regardless of the symptoms.


LncRNA GUARDIN suppresses cellular senescence through a LRP130-PGC1α-FOXO4-p21-dependent signaling axis.
The long noncoding RNA GUARDIN functions to protect genome stability. Inhibiting GUARDIN expression can alter cell fate decisions toward senescence or apoptosis, but the underlying molecular signals are unknown. Here, we show that GUARDIN is an essential component of a transcriptional repressor complex involving LRP130 and PGC1α. GUARDIN acts as a scaffold to stabilize LRP130/PGC1α heterodimers and their occupancy at the FOXO4 promotor. Destabilizing this complex by silencing of GUARDIN, LRP130, or PGC1α leads to increased expression of FOXO4 and upregulation of its target gene p21, thereby driving cells into senescence. We also found that GUARDIN expression was induced by rapamycin, an agent that suppresses cell senescence. FOS-like antigen 2 (FOSL2) acts as a transcriptional repressor of GUARDIN, and lower FOSL2 levels in response to rapamycin correlate with increased levels of GUARDIN. Together, these results demonstrate that GUARDIN inhibits p21-dependent senescence through a LRP130-PGC1α-FOXO4 signaling axis, and moreover, GUARDIN contributes to the anti-aging activities of rapamycin.


Predictors of Intubation in Patients With Acute Hypoxemic Respiratory Failure Treated With a Noninvasive Oxygenation Strategy.
In patients with acute hypoxemic respiratory failure, noninvasive ventilation and high-flow nasal cannula oxygen are alternative strategies to conventional oxygen therapy. Endotracheal intubation is frequently needed in these patients with a risk of delay, and early predictors of failure may help clinicians to decide early. We aimed to identify factors associated with intubation in patients with acute hypoxemic respiratory failure treated with different noninvasive oxygenation techniques. Post hoc analysis of a randomized clinical trial. Twenty-three ICUs. Patients with a respiratory rate greater than 25 breaths/min and a PaO2/FIO2 ratio less than or equal to 300 mm Hg. Patients were treated with standard oxygen, high-flow nasal cannula oxygen, or noninvasive ventilation. Respiratory variables one hour after treatment initiation. Under standard oxygen, patients with a respiratory rate greater than or equal to 30 breaths/min were more likely to need intubation (odds ratio, 2.76; 95% CI, 1.13-6.75; p = 0.03). One hour after high-flow nasal cannula oxygen initiation, increased heart rate was the only factor associated with intubation. One hour after noninvasive ventilation initiation, a PaO2/FIO2 ratio less than or equal to 200 mm Hg and a tidal volume greater than 9 mL/kg of predicted body weight were independent predictors of intubation (adjusted odds ratio, 4.26; 95% CI, 1.62-11.16; p = 0.003 and adjusted odds ratio, 3.14; 95% CI, 1.22-8.06; p = 0.02, respectively). A tidal volume above 9 mL/kg during noninvasive ventilation remained independently associated with 90-day mortality. In patients with acute hypoxemic respiratory failure breathing spontaneously, the respiratory rate was a predictor of intubation under standard oxygen, but not under high-flow nasal cannula oxygen or noninvasive ventilation. A PaO2/FIO2 below 200 mm Hg and a high tidal volume greater than 9 mL/kg were the two strong predictors of intubation under noninvasive ventilation.


Spindle birefringence of isolated mitotic apparatus: further evidence for two birefringent spindle components.
We studied sea-urchin zygote mitotic apparatus (MA) isolated in hexylene glycol, transferred immediately to a glycerol-dimethylsulphoxide medium, and stored for 2 weeks at room temperature. Treatment with 0-5 M KC1 caused loss of 45% of the birefringence, but microtubules remained intact (as seen electron microscopically in glutaraldehyde-fixed MA), and tubulin was not extracted (as determined by polyacrylamide gel electrophoresis). These results suggest that a non-tubulin component which is extracted by the KC1 contributes 45% of the MA birefringence. Further evidence for this conclusion came from indirect immunofluorescence experiments. Non-extracted (control) MA were fixed with formaldehyde and reacted with antibody against tubulin; there was intense staining of the spindle fibres and astral rays. Electron microscopically, however, microtubules were not present in formaldehyde-fixed MA. Since formaldehyde fixation caused breakdown of microtubules but the tubulin remained in the MA (as judged by reaction with antibodies) we suggest that after microtubule breakdown the tubulin remains in the MA because it is bound to a peri-microtubule spindle component (which we call 'substance gamma'). When KCl-extracted MA were fixed with formaldehyde and reacted with antibody against tubulin there was very little staining of spindle fibres and astral rays. Electron microscopically, formaldehyde caused microtubule breakdown, and since the tubulin is lost from formaldehydefixed, KC1-extracted MA (as judged by reaction with antibodies), we suggest that the tubulin-binding component, substance gamma, is extracted by the 0-5 M KC1. Pressure treatment caused the asters not to stain with antibody against tubulin, suggesting that the stability of substance gamma is different in different regions of the mitotic apparatus.