Reactivation of interference during sleep does not impair ongoing memory consolidation.
Memory consolidation during sleep is assumed to rely on the repeated reactivation of newly encoded memories particularly during slow wave sleep (SWS). It has been proposed that reactivated memories during sleep - like during wakefulness - undergo a labilisation process, enabling the strengthening and integration of new memories into pre-existing networks. Here, we tested this idea by introducing interference directly during sleep in the reactivation/consolidation phase. We predicted that cueing interfering memories during sleep would impair the consolidation of recently learned memories. Participants learned a visuo-spatial memory task before they were allowed to sleep for 40 min. During sleep, and particularly during SWS, subjects were presented with interference via odour cueing (compared to a no interference vehicle condition). In contrast to our hypothesis, cueing of the interference during sleep did not impair consolidation of the newly learned memories: odour and vehicle conditions did not differ in memory recall after sleep. On the contrary, subjects even displayed significantly fewer intrusions from the interference during memory recall when the odour was presented during sleep. These findings suggest that interference presentation during sleep does not disrupt endogenous memory consolidation, but might even facilitate pattern separation and memory stabilisation through generalisation processes.


Airway hyper-responsiveness in lipopolysaccharide-challenged common marmosets (Callithrix jacchus).
Animal models with a high predictive value for human trials are needed to develop novel human-specific therapeutics for respiratory diseases. The aim of the present study was to examine lung-function parameters in marmoset monkeys (Callithrix jacchus) that can be used to detect pharmacologically or provocation-induced AHR (airway hyper-responsiveness). Therefore a custom-made lung-function device that allows application of defined aerosol doses during measurement was developed. It was hypothesized that LPS (lipopolysaccharide)-challenged marmosets show AHR compared with non-challenged healthy subjects. Invasive plethysmography was performed in 12 anaesthetized orotracheally intubated and spontaneously breathing marmosets. Pulmonary data of R(L) (lung resistance), C(dyn) (dynamic compliance), EF50 (mid-expiratory flow), P(oes) (oesophageal pressure), MV (minute volume), respiratory frequency (f) and V(T) (tidal volume) were collected. Measurements were conducted under baseline conditions and under MCh (methacholine)-induced bronchoconstriction. The measurement was repeated with the same group of animals after induction of an acute lung inflammation by intratracheal application of LPS. PDs (provocative doses) of MCh to achieve a certain increase in RL were significantly lower after LPS administration. AHR was demonstrated in the LPS treated compared with the naïve animals. The recorded lung-function data provide ground for pre-clinical efficacy and safety testing of anti-inflammatory substances in the common marmoset, a new translational NHP (non-human primate) model for LPS-induced lung inflammation.


Mesoscale conformational changes in the DNA-repair complex Rad50/Mre11/Nbs1 upon binding DNA.
The human Rad50/Mre11/Nbs1 complex (hR/M/N) functions as an essential guardian of genome integrity by directing the proper processing of DNA ends, including DNA breaks. This biological function results from its ability to tether broken DNA molecules. hR/M/N's dynamic molecular architecture consists of a globular DNA-binding domain from which two 50-nm-long coiled coils protrude. The coiled coils are flexible and their apices can self-associate. The flexibility of the coiled coils allows their apices to adopt an orientation favourable for interaction. However, this also allows interaction between the tips of two coiled coils within the same complex, which competes with and frustrates the intercomplex interaction required for DNA tethering. Here we show that the dynamic architecture of hR/M/N is markedly affected by DNA binding. DNA binding by the hR/M/N globular domain leads to parallel orientation of the coiled coils; this prevents intracomplex interactions and favours intercomplex associations needed for DNA tethering. The hR/M/N complex thus is an example of a biological nanomachine in which binding to its ligand, in this case DNA, affects the functional conformation of a domain located 50 nm distant.


The separation of alpha-2 macroglobulin into five components with differing electrophoretic and enzyme-binding properties.
The alpha-2 macroglobulins from human serum and plasma were isolated by Bio-Gel P-300 and A5m gel filtration. The material showed a single peak on sedimentation velocity ultracentrifugation, a mol wt of 650,000 by sedimentation equilibrium ultracentrifugation, and a major precipitin arc in the alpha-2 macroglobulin region by immunoelectrophoresis against whole human serum. Two bands were observed in the alpha-2 macroglobulin region when acrylamide gel electrophoresis was performed with a pH 8.9 running gel. When a pH 7.8 gel was used, five electrophoretic species were observed. In both cases, the preaddition of stoichiometric amounts of trypsin or chymotrypsin added to alpha-2 macroglobulin resulted in disappearance of slower bands leaving only one band on acrylamide gel electrophoresis patterns. Preparative acrylamide gel electrophoresis separated alpha-2 macroglobulin obtained from Bio-Gel into five closely-spaced species. Separation was sufficiently adequate to show that those species of alpha-2 macroglobulin which bound trypsin and chymotrypsin were represented by slower moving species and that the fastest moving material had lost virtually all of the ability to bind these enzymes. Preparative acrylamide gel electrophoresis of a mixture of alpha-2 macroglobulin-trypsin complex and alpha-2 macroglobulin revealed that the fast moving component was alpha-2 macroglobulin-trypsin complex and that the slower moving material was unbound alpha-2 macroglobulin. The naturally occurring amidase activity of the alpha-2 macroglobulin using benzoylarginine-p-nitroanilide (BAPNA) as substrate was investigated and unlike its trypsin-binding activity, amidase activity was found to be of the same specific activity in all electrophoretic fractions. Binding of trypsin and chymotrypsin to alpha-2 macroglobulin revealed that alpha-2 macroglobulin maximally bound 2 moles of trypsin and 1 mole of chymotrypsin. When the enzymes were added simultaneously there was competition. Chymotrypsin added to alpha-2 macroglobulin before the addition of trypsin prevented all trypsin binding even though only one site was filled with chymotrypsin. These results were explained by the acrylamide gels which showed that 1 mole of chymotrypsin was sufficient to convert all the alpha-2 macroglobulin to a species with the fastest mobility which no longer binds additional enzyme.


Heat shock protein 47 expression in aged normal human fibroblasts: modulation by Salix alba extract.
Heat shock protein (HSP) 47 is a specific chaperone of procollagen. This heat shock protein is responsible for the correct three-dimensional organization of procollagen and its control-quality prior secretion. The aim of the study is to evaluate the level of HSP 47 in aged, photoaged, and senescent fibroblasts and its modulation by a plant extract (Salix alba). The level of HSP 47 and/or procollagen expression in fibroblasts was measured by real-time RT-PCR (mRNA transcripts) and by flow cytometry (immunochemistry technique for measurement of arbitrary fluorescence intensity). Immunochemistry techniques and confocal microscopy were used to visualize the cellular localization of HSP 47 and procollagen. These parameters were compared with different age donors, nonsenescent, and senescent fibroblasts. Fibroblasts were irradiated by a noncytotoxic dose of UVA (6 J/cm(2)), and HSP 47 level was evaluated. S. alba extract was tested for its capacity to modulate HSP 47 expression. Colocalization of HSP 47 and procollagen was shown by confocal microscopy, indicating that HSP 47 could play a role of procollagen molecular chaperone in the cellular model. It was also shown that the HSP 47 level is decreased in old-donor cells, senescent, and irradiated cells. This decrease can be modulated by a S. alba extract (polyphenols rich) in a dose-dependent manner. The evaluation of HSP 47 expression in the experimental conditions can lead to a new approach of aging and photoaging, pointing out the implication of this chaperone in these pathophysiologic phenomena. Modulation of HSP 47 expression by this family of molecules could be of cosmetic and/or dermatologic interest.


Serum Uric Acid Level as a Prognostic Marker in Patients With Acute Respiratory Distress Syndrome.
Uric acid acts as both a pathogenic inflammatory mediator and an antioxidative agent. Several studies have shown that uric acid level correlates with the incidence, severity, and prognosis of pulmonary diseases. However, the association between uric acid level and acute respiratory distress syndrome (ARDS) has not been studied. This study was conducted to elucidate how serum uric acid level is related with clinical prognosis of ARDS. A retrospective cohort study with propensity score matching was conducted at a medical intensive care unit of a tertiary teaching hospital. The medical records of patients diagnosed with ARDS admitted from 2005 through 2011 were reviewed. Two hundred thirty-seven patients with ARDS met the inclusion criteria. Patients with a serum uric acid level <3.0 mg/dL were classified into the low uric acid group, and those with a level ≥3 mg/dL were classified into the normal to high uric acid group. We selected 40 patients in each group using propensity score matching. A higher percentage of patients in the low uric acid group experienced clinical improvement in ARDS. More patients died from sepsis in the normal to high uric acid group. Kaplan-Meier analysis showed that a low serum uric acid level was significantly associated with better survival rate. In patients with ARDS, a low serum uric acid level may be a prognostic marker of a low risk of in-hospital mortality.


Hydrostatic pressure stimulation of human mesenchymal stem cells seeded on collagen-based artificial extracellular matrices.
Human mesenchymal stem cells (hMSCs) from bone marrow are considered a promising cell source for bone tissue engineering applications because of their ability to differentiate into cells of the osteoblastic lineage. Mechanical stimulation is able to promote osteogenic differentiation of hMSC; however, the use of hydrostatic pressure (HP) has not been well studied. Artificial extracellular matrices containing collagen and chondroitin sulfate (CS) have promoted the expression of an osteoblastic phenotype by hMSCs. However, there has been little research into the combined effects of biochemical stimulation by matrices and simultaneous mechanical stimulation. In this study, artificial extracellular matrices generated from collagen and/or CS were coated onto polycaprolactone-co-lactide substrates, seeded with hMSCs and subjected to cyclic HP at various time points during 21 days after cell seeding to investigate the effects of biochemical, mechanical, and combined biochemical and mechanical stimulations. Cell differentiation was assessed by analyzing the expression of alkaline phosphatase (ALP) at the protein- and mRNA levels, as well as for calcium accumulation. The timing of HP stimulation affected hMSC proliferation and expression of ALP activity. HP stimulation after 6 days was most effective at promoting ALP activity. CS-containing matrices promoted the osteogenic differentiation of hMSCs. A combination of both CS-containing matrices and cyclic HP yields optimal effects on osteogenic differentiation of hMSCs on scaffolds compared with individual responses.


[High prevalence and incidence of hepatitis C virus infections among dialysis patients in the East-Centre of Tunisia].
Hemodialysed patients are recognised as a group at increased risk of infection with hepatitis C virus (HCV). The aim of this study was to determine the prevalence and incidence of HCV infection among dialysis patients of the east-centre part of Tunisia. Two hundred and seventy-six patients dialysed until 2001 were recruited within seven hemodialysis units located in the cities of Sousse, Monastir and Mahdia. The serum markers of HCV infection were tested over the period of March 2000-December 2002, by a 3rd generation ELISA test for antibodies and by qualitative RT-PCR technique for viral RNA. The prevalence of anti-HCV antibodies and of HCV RNA was 32.6% (90 patients) and 25.7% (71 patients), respectively. Between 1998 and 2002, 20 new infections were documented in five of the seven dialysis units corresponding to an incidence of 2.34% per year, with an average time of contamination after the beginning of dialysis of 4.6 years. If all the infections are assessed to have occurred during dialysis, the density of incidence of HCV contamination was 4.4% per year of dialysis. A high correlation was noticed between the presence of HCV markers in serum and the duration of dialysis (F = 34.15, P < 0.0001). In the absence of other risk factors (transfusion, drug-addiction), these results plead for the nosocomial transmission of the observed HCV infections. A phylogenetic analysis of the E2 hypervariable region of the viral genome is in progress to confirm this assumption.


Phospholipase D in TCR-Mediated Signaling and T Cell Activation.
Phospholipase D (PLD) is an enzyme that catalyzes the hydrolysis of phosphatidylcholine, the major phospholipid in the plasma membrane, to generate an important signaling lipid, phosphatidic acid. Phosphatidic acid is a second messenger that regulates vesicular trafficking, cytoskeletal reorganization, and cell signaling in immune cells and other cell types. Published studies, using pharmacological inhibitors or protein overexpression, indicate that PLD plays a positive role in TCR-mediated signaling and cell activation. In this study, we used mice deficient in PLD1, PLD2, or both to assess the function of these enzymes in T cells. Our data showed that PLD1 deficiency impaired TCR-mediated signaling, T cell expansion, and effector function during immune responses against Listeria monocytogenes; however, PLD2 deficiency had a minimal impact on T cells. Biochemical analysis indicated that PLD1 deficiency affected Akt and PKCθ activation. In addition, it impaired TCR downregulation and the secondary T cell response. Together, our results suggested that PLD1 plays an important role in T cell activation.


Why Travel for Complex Cancer Surgery? Americans React to 'Brand-Sharing' Between Specialty Cancer Hospitals and Their Affiliates.
Leading cancer hospitals have increasingly shared their 'brand' with smaller hospitals through affiliations. Because each brand evokes a distinct reputation for the care provided, 'brand-sharing' has the potential to impact the public's ability to differentiate the safety and quality within hospital networks. The general public was surveyed to determine the perceived similarities and differences in the safety and quality of complex cancer surgery performed at top cancer hospitals and their smaller affiliate hospitals. A national, web-based KnowledgePanel (GfK) survey of American adults was conducted. Respondents were asked about their beliefs regarding the quality and safety of complex cancer surgery at a large, top-ranked cancer hospital and a smaller, local hospital, both in the presence and absence of an affiliation between the hospitals. A total of 1010 surveys were completed (58.1% response rate). Overall, 85% of respondents felt 'motivated' to travel an hour for complex surgery at a larger hospital specializing in cancer, over a smaller local hospital. However, if the smaller hospital was affiliated with a top-ranked cancer hospital, 31% of the motivated respondents changed their preference to the smaller hospital. When asked to compare leading cancer hospitals and their smaller affiliates, 47% of respondents felt that surgical safety, 66% felt guideline compliance, and 53% felt cure rates would be the same at both hospitals. Approximately half of surveyed Americans did not distinguish the quality and safety of surgical care at top-ranked cancer hospitals from their smaller affiliates, potentially decreasing their motivation to travel to top centers for complex surgical care.


A review of organizational culture instruments for nurse executives.
Nurse executives are aware of the complexities of organizational culture. It impacts the nursing work environment and patient care safety and quality. The authors describe several widely available tools that nurse leaders can use to assess organizational culture in the work environment. The psychometric and conceptual strengths and weaknesses of the measures are described and recommendations for use in nursing and patient care administration are provided.


EBV-associated Burkitt lymphoma in Taiwan is not age-related.
This study retrospectively investigated 54 cases of sporadic Burkitt lymphoma in Taiwan with histopathology review, immunohistochemistry, Epstein-Barr virus (EBV) in situ hybridization (EBER) and fluorescence in situ hybridization (FISH). The great majority revealed typical immunophenotype and 89% (47/53) cases expressed myc protein. EBER was positive in 20% (11/54) of cases, more frequently with nodal presentation, but not significantly associated with age (pediatric vs. adult), abdominal vs. extra-abdominal presentation or overall survival (OS). MYC and IGH were rearranged in 94% (46/49) and 85% (41/48) of cases, respectively. The concordance rate between myc expression and MYC translocation was 83% (40/48). By univariate analysis, OS was statistically associated with age, with or without chemotherapy, central nervous system (CNS) involvement, CNS prophylaxis and leukemic transformation, but not gender, nodal vs. extranodal involvement, stage, immunohistochemistry, EBER, myc expression, MYC translocation or radiotherapy. By multivariate analysis, CNS involvement at presentation and administration of chemotherapy were statistically associated with OS.


Staged arthrodesis for salvage of the septic hallux metatarsophalangeal joint.
Failed surgery at the hallux metatarsophalangeal joint may present substantial difficulties in treatment, especially when complicated by infection. This retrospective study reviews the staged treatment of 5 patients with complications of hallux valgus surgery associated with sepsis of the metatarsophalangeal joint. The initial salvage treatment included debridement with placement of an antibiotic cement spacer and either an intramedullary Kirschner wire (4 patients) or an external fixator (1 patient). At the second stage procedure, the spacer and fixation were removed, and an autogenous tricortical iliac crest graft was inserted into the joint. All patients had clinical control of joint sepsis. Pseudoarthrosis occurred at the proximal end of the tricortical graft in 2 patients. Patients were evaluated at an average of 28 months after the fusion. Staged arthrodesis appears to be a satisfactory approach to postoperative sepsis of the hallux metatarsophalangeal joint.


Temporal trends and predictors of inhospital death in patients hospitalised for heart failure in Germany.
We investigated trends in incidence, case fatality rate, patient characteristics and adverse inhospital events of patients hospitalised for heart failure in Germany. The German nationwide inpatient sample (2005-2016) was used for this analysis. Patients hospitalised due to heart failure were selected for analysis. Temporal trends in the incidence of hospitalisations, case fatality rate and treatments were analysed and predictors of inhospital death were identified. The analysis comprised a total number of 4,539,140 hospitalisations (52.0% women, 81.0% aged ≥70 years) due to heart failure. Although hospitalisations increased from 381 (2005) to 539 per 100,000 population (2016) (β estimate 0.06, 95% confidence interval (CI) 0.06 to 0.07, P < 0.001) in parallel with median age and prevalence of comorbidities, the inhospital case fatality rate decreased from 11.1% to 8.1% (β estimate -0.36, 95% CI -0.37 to -0.35, P < 0.001) and the rate of major adverse cardiovascular and cerebrovascular events (β estimate -0.24, 95% CI -0.25 to -0.23, P < 0.001) decreased from 12.7% to 10.3%. Age 70 years and older (odds ratio (OR) 2.60, 95% CI 2.57 to 2.63, P < 0.001) and cancer (OR 1.93, 95% CI 1.91 to 1.96, P < 0.001) were independent predictors of inhospital death. Hospitalisations for heart failure increased in Germany from 2005 to 2016, whereas the major adverse cardiovascular and cerebrovascular event rate and inhospital case fatality rate decreased during this period despite higher patient age and increasing prevalence of comorbidities.


New species of Dugdaleiella, gen. nov., Kozloviella, gen. nov., and Pfitzneriella Viette from upper elevation Andes of Ecuador and Peru (Lepidoptera: Hepialidae).
Six new Hepialidae species are described from high elevation habitats in the Andes of Peru and Ecuador. One species is assigned to the genus Kozloviella gen. n., K. viazmenskyi sp. n., from Peru, and five to the genus Pfitzneriella Viette, 1951: P. antonkozlovi sp. n., P. olafi sp. n., P. titarenkoi, sp. n., P. yuliyakovalevae sp. n. from Peru and P. rawlinsi sp. n., from Ecuador. Dugdaleiella gen. n., is proposed to accommodate the Ecuadorian species Dugdaleiella monticola (Maassen, 1890) comb. n., formerly placed in Pfitzneriella. The taxonomic (but not the phylogenetic) status of P. lucicola (Maassen, 1890) is unaltered pending future opportunity to examine the type series. Preliminary phylogenetic analysis of the morphological characters supports Kozloviella gen. n., and Pfitzneriella as sister taxa, and separates P. titarenkoi sp. n., from the other species of Pfitzneriella. We suggest that the ancestor of these high elevation genera already occupied a pre-Andean upland habitat across Peru and Ecuador and this ancestor further diverged into the modern genera and species during the Andean orogeny. We consider it likely that there is a considerable diversity of endemic high elevation Hepialidae that remains to be discovered in the eastern Andes between Venezuela and Bolivia.


Modeling Dose Response for Late Dysphagia in Patients With Head and Neck Cancer in the Modern Era of Definitive Chemoradiation.
To develop personalized multivariate dose-response models for late dysphagia in patients with head and neck cancer treated in the modern era of combined chemotherapy with intensity-modulated radiation therapy. The analysis included 424 patients (oropharyngeal cancer [n = 295] and nasopharyngeal, hypopharyngeal, or laryngeal cancer [n = 129]) who received definitive chemoradiation between January 2004 and April 2009. The superior, middle, and inferior pharyngeal constrictor muscles were contoured. We calculated generalized equivalent uniform dose (gEUD) for each and the total constrictor muscle volume, with the volume effect parameter a varying from log10 a = -1 to +1 in steps of 0.1. We used the National Cancer Institute Common Toxicity Criteria for Adverse Events (version 3.0) to grade late dysphagia and logistic regression to evaluate the correlation of gEUD( a) with grade 2 or higher (≥ G2) and grade 3 or higher (≥ G3) late dysphagia at each value of a. Median follow-up was 33.3 months (range, 6 to 69 months). There were 41 cases (10%) of ≥ G2 dysphagia and 22 cases (5%) of ≥ G3 dysphagia. Mean doses to the total constrictor ranged from 30.1 to 85.7 Gy (median, 61.2 Gy). The predicted rate of ≥ G2 dysphagia increased by approximately 3.4% per Gy at the mean dose, for which the probability of ≥ G2 dysphagia is 50%. The threshold mean total constrictor doses that limited rates of ≥ G2 and ≥ G3 dysphagia to < 5% were < 58 Gy and < 61 Gy, respectively. Other significant factors in the multivariate predictive model included disease site, mean dose to total constrictor muscle, and patient age. Incidences of both ≥ G2 and ≥ G3 dysphagia were dependent on the mean radiation dose to the total constrictor muscle volume, disease site, and patient age. Limiting the total volume of constrictor muscle to < 58 Gy could keep the predicted rate of ≥ G2 dysphagia to < 5%.


A differential hybridization scheme to identify photoreceptor-specific genes.
Identification of genes expressed preferentially or exclusively in photoreceptors will facilitate the understanding of photoreceptor biology as well as provide candidate genes for inherited retinal degenerations. To achieve this goal we performed a differential hybridization screen of 3717 well-isolated phage clones from a human retinal cDNA library. Clones were selected for further study if they hybridized exclusively or strongly preferentially to a probe derived from RNA isolated from the cone-predominant retina of 13-line ground squirrels as compared to a probe derived from human fibroblast RNA. Twenty percent of clones (9/45) identified by this screen were derived from photoreceptor-specific genes and an additional 24.4% (11/45) were from neural-specific genes, demonstrating the utility of this strategy in identifying genes important for retinal biology.


Efficacy of a 0.1% capsaicin hydrogel patch for myofascial neck pain: a double-blinded randomized trial.
The objective of this study was to evaluate the efficacy of a hydrogel patch containing capsaicin 0.1% compared with a placebo hydrogel patch without capsaicin to treat chronic myofascial neck pain. The study was designed as a double-blinded randomized controlled trial. Setting. The study was set at Kyung-hee University Hospital at Gangdong, Korea. Sixty-one participants between 18 and 65 years with at least 3 months duration of neck pain and a clinical presentation of myofascial pain syndrome were enrolled in the study from September 1 to November 20, 2010. Interventions. Participants received capsaicin 0.1% hydrogel patches or control hydrogel patches without capsaicin according to the randomization scheme. All participants were instructed to apply one patch to each side of the neck and shoulder girdle overlying the point of maximal pain for 12 hours daily during the duration of the 4-week study. Each participant completed five surveys at baseline, at 2 weeks after the start of treatments, and at the conclusion of the 4-week study. The primary outcome measure was visual analog scale (VAS). Other outcome measures included the Neck Disability Index (NDI), Beck's Depression inventory (BDI), Short Form 36 Korean version, and Euroqol 5-D. Fifty-seven patients completed the study. The mean VAS, NDI, and BDI scores were significantly decreased at 2 and 4 weeks after the start of the intervention in both groups. There was no significant difference between the two groups in any of the outcome measures. Future research may help to discern specific effects of capsaicin, trigger point stimulation by application of the patch, and the placebo effect.


Low-density lipoproteins induce heat shock protein 27 dephosphorylation, oligomerization, and subcellular relocalization in human vascular smooth muscle cells.
High levels of circulating low-density lipoproteins (LDL) are a major atherosclerotic risk factor. The effects of intimal LDL on vascular smooth muscle cell (VSMC) phenotype and function during plaque remodeling and vascular repair are not fully understood. We have investigated whether exposure of VSMC to LDL induces changes on the proteomic profile of the heat shock protein (HSP) family-molecular chaperones involved in atherosclerosis. Methods & Results- 2D electrophoresis demonstrates that LDL modifies the proteomic profile of HSP27 (HSPB1). Western blot analysis evidenced a significant HSP27 dephosphorylation after exposure of cells to native LDL (nLDL) and aggregated-LDL (agLDL) for 24 hours (P<0.05). Dephosphorylation of HSP27 was not related to changes in p38(MAPK) phosphorylation. Both nLDL and agLDL induced relocalization of unphosphorylated HSP27 to the tip of actin stress fibers and focal adhesion structures in VSMC. During cell adhesion, phospho-HSP27 was located at the cell surface contact region in LDL-treated cells, whereas it remained cytosolic in control cells. Immunohistochemistry studies showed that phosphorylated HSP27 is rarely found in lipid-rich areas of atherosclerotic plaques in human coronary arteries. Our results indicate that in VSMC, LDL modulate HSP27 phosphorylation and subcellular localization, affecting actin polymerization and cytoskeleton dynamics.


Long-term use of inhaled corticosteroids and risk of upper respiratory tract infection in chronic obstructive pulmonary disease: a meta-analysis.
Recent studies have suggested that inhaled corticosteroids (ICS) may be associated with higher risks of tuberculosis and pneumonia in patients with COPD. However, it is not known whether ICS increases the risk of upper respiratory tract infection (URTI). Aim of this study was to explore the relationship between ICS and URTI. Through a comprehensive literature search of PubMed, EMBASE, Cochrane Library, and Google Scholar from inception to March 2016, we identified randomized controlled trials of ICS therapy lasting at least 6 months. A meta-analysis by the Peto approach was also conducted to generate summary estimates comparing ICS with non-ICS treatment on the risk of URTI. A total of 14 studies involving 19,777 subjects were considered in the meta-analysis. Compared with non-ICS treatment, ICS were associated with a significantly increased risk of URTI (Peto OR: 1.16; 95% CI: 1.05-1.29; I2 = 9%; p = .004). Subgroup analyzes were performed for different dose, high-dose ICS was associated with a significantly increased risk of URTI (Peto OR: 1.19; 95% CI: 1.05-1.34; I2 = 0%; p = .005), whereas low-dose ICS showed a non-significant increased risk of URTI (Peto OR: 1.10; 95% CI: 0.91-1.33; I2 = 0%; p = .32). Moreover, fluticasone was observed with an increased risk of URTI but not mometasone; high-dose fluticasone treatment was associated with a significantly higher risk of URTI but not low-dose. These results suggested to us that ICS use may increase the risk of URTI in patients with COPD, but it should be further investigated.


Identification, Characterization, and Formulation of a Novel Carbapenemase Intended to Prevent Antibiotic-Mediated Gut Dysbiosis.
Antibiotics can damage the gut microbiome leading to opportunistic infections and the emergence of antibiotic resistance. Microbiome protection via antibiotic inactivation in the gastrointestinal (GI) tract represents a strategy to limit antibiotic exposure of the colonic microbiota. Proof of concept for this approach was achieved with an orally-administered beta-lactamase enzyme, SYN-004 (ribaxamase), that was demonstrated to degrade ceftriaxone excreted into the GI tract and protect the gut microbiome from antibiotic-mediated dysbiosis. Ribaxamase efficiently degrades penicillin and cephalosporin beta-lactam antibiotics, but is not active against carbapenems. To expand this microbiome protection strategy to include all classes of beta-lactams, three distinct carbapenemases were evaluated for manufacturability, antibiotic degradation spectrum, and stability in human intestinal fluid. E. coli production strains were generated for P2A, a novel metallo-enzyme isolated from B. cereus, New Delhi metallo-beta-lactamase (NDM), and Klebsiella pneumoniae carbapenemase (KPC). While all three enzymes effectively inactivated a broad range of antibiotics, including penicillins, most cephalosporins, and carbapenems in vitro, only P2A retained biological activity when incubated with human chyme. As functional stability in the intestinal tract is a key requirement for an orally-delivered enzyme, P2A was chosen as a potential clinical candidate. An enteric formulation of P2A was developed, called SYN-006, that was inert under high acid conditions, with enzyme dissolution occurring at pH > 5.5. SYN-006 has the potential to expand microbiome protection via antibiotic inactivation to include all classes of beta-lactam antibiotics.


Competitive amperometric immunosensor based on covalent linking of a protein conjugate to dendrimer-functionalised nanogold substrate for the determination of 2,4,6-trinitrotoluene.
A new amperometric immunosensor for 2,4,6-trinitrotoluene based on the working principle of competitive enzyme-linked immunosorbent assay was developed and characterised. An electrodeposited nanogold substrate was functionalised by deposition of self-assembled monolayers of 2-aminoethanethiol as linkers for the subsequent immobilisation of polyamidoaminic dendrimers. Our approach makes use of those dendrimers to anchor a trinitrobenzene-ovalbumin conjugate on the electrode surface. The immunosensor was tested and validated for the determination of 2,4,6-trinitrotoluene showing high selectivity with respect to other nitroaromatic compounds, a limit of detection of 4.8 ng/mL and a limit of quantitation of 6 ng/mL. The immunosensor was tested for the quantification of the analyte in spiked soils and in a real sample of post-blast soil, evidencing a good recovery rate (113 %).


Mussel Consumption as a "Food First" Approach to Improve Omega-3 Status.
Numerous United Kingdom and European Union expert panels recommend that the general adult population consumes ~250 mg of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) per day through the consumption of one portion of oily fish per week. The long-chain omega-3 fatty acids EPA and DHA are only found in appreciable amounts in marine organisms. Increasing oily fish consumption conflicts with sustaining fisheries, so alternative dietary sources of EPA and DHA must be explored. Mussels are high in omega-3 polyunsaturated fatty acids (PUFAs) and a good source of essential amino acids. Therefore, we aimed to investigate the impact of introducing mussels as a protein source in the lunchtime meal three times per week for two weeks on the omega-3 status of free-living participants. Following an initial two-week monitoring period, 12 participants (eight male and four female) attended the nutrition laboratory three times per week for two weeks. Each participant received a personalised lunch constituting one-third of their typical daily calorie consumption with ~20% of the calories supplied as cooked mussels. A portion of cooked mussels from each feeding occasion was tested for total omega-3 content. The mean ± SD mussel EPA + DHA content was 518.9 ± 155.7 mg/100 g cooked weight, meaning that each participant received on average 709.2 ± 252.6 mg of EPA + DHA per meal or 304.0 ± 108.2 mg of EPA + DHA per day. Blood spot analysis revealed a significant increase in the omega-3 index (week 1 = 4.27 ± 0.81; week 4 = 5.07 ± 1.00) and whole blood EPA content during the study (%EPA week 1 = 0.70 ± 0.0.35; %EPA week 4 = 0.98 ± 0.35). Consuming mussels three times per week for two weeks as the protein source in a personalised lunchtime meal is sufficient to moderately improve the omega-3 index and whole blood DHA + EPA content in young healthy adults.


[Histoplasmosis updating].
Histoplasmosis, an infection caused by the fungus Histoplasma capsulatum, has been reported all over the world and is considered endemic in the American continent, including Cuba. This fungus grows on the soils contaminated with bird and bat excreta, where it produces a great number of microconidia that could cause the infection when they are inhaled. The clinical spectrum varies from asymptomatic infections to serious disseminated diseases involving one or many organ systems and affects mainly AIDS patients, patients with hematological neoplasias, transplant recipients or other immunosuppressed patients. The main risk groups include those individuals whose working activities make them be exposed to aerosols contaminated with H. capsulatum. Lab diagnosis is based on the microscopic observation, isolation and identification of the fungus in fluid or tissue samples of patients, and on specific antigen and antibodies detection. The molecular methods based on polymerase chain reaction have not been sufficiently defined, and they could be an important advance in the early diagnosis of this mycosis. Itraconazole is recommended for treatment of moderate, localized and chronic infection whereas amphotericin B is the drug of choice for disseminated and serious manifestations, particularly in its lipidic formulations. At present, histoplasmosis is considered one of the most important systemic mycoses in the Americas, and it is widely spread over all regions of Cuba.


Chemical modification of the lactose carrier of Escherichia coli by plumbagin, phenylarsinoxide or diethylpyrocarbonate affects the binding of galactoside.
The effects of chemical modification of the lactose carrier of Escherichia coli on galactoside binding (in overproducing strains) and on transport was examined. Both the modifying reagents diethylpyrocarbonate and rose bengal and the thiol reagents phenylarsinoxide and plumbagin can completely inhibit the binding of the substrate p-nitrophenyl alpha-D-galactopyranoside to the carrier. If care is taken to inhibit galactoside binding only partially, the loss of transport is found to parallel the loss of binding sites. The modified carrier molecules are completely inactive, while the remaining active carrier molecules evince normal transport and binding parameters. The binding of galactoside protects the carrier partially against these forms of chemical modification. In view of these observations, the results of previous chemical modification studies [Padan, E., Patel, L. and Kaback, H.R. (1979) Proc. Natl Acad. Sci. USA, 76, 6221-6225; Konings, W.N. and Robillard, G.T. (1982) Proc. Natl Acad. Sci. USA, 79, 5480-5484] must be re-interpreted. Our results stress the utility of studying substrate binding, the first step in the transport cycle.


The role of nutrition in the pathophysiology and management of sickle cell disease among children: A review of literature.
Sickle cell disease (SCD) is one of the common inherited blood disorders in humans and has been associated with decreased dietary intake which results in poor nutritional status and impaired growth. Nutrition is one of the most important but often forgotten aspect of care of patients with chronic disorders and there have been emerging concern in literature on increased nutritional needs of SCD patients. This paper sought to review the available literature on the roles of individual nutrients in the pathophysiology and management of SCD among children. Children with SCD have been shown to exhibit suboptimal status with respect to both macronutrients and micronutrients. Thus, nutrition could play an important role in the management of SCD. However, there is paucity of evidence coming from trials with large sample sizes to support the suggestion that supplementation with various nutrients that have been considered in this review will be helpful.


Kinases Mst1 and Mst2 positively regulate phagocytic induction of reactive oxygen species and bactericidal activity.
Mitochondria need to be juxtaposed to phagosomes for the synergistic production of ample reactive oxygen species (ROS) in phagocytes to kill pathogens. However, how phagosomes transmit signals to recruit mitochondria has remained unclear. Here we found that the kinases Mst1 and Mst2 functioned to control ROS production by regulating mitochondrial trafficking and mitochondrion-phagosome juxtaposition. Mst1 and Mst2 activated the GTPase Rac to promote Toll-like receptor (TLR)-triggered assembly of the TRAF6-ECSIT complex that is required for the recruitment of mitochondria to phagosomes. Inactive forms of Rac, including the human Rac2(D57N) mutant, disrupted the TRAF6-ECSIT complex by sequestering TRAF6 and substantially diminished ROS production and enhanced susceptibility to bacterial infection. Our findings demonstrate that the TLR-Mst1-Mst2-Rac signaling axis is critical for effective phagosome-mitochondrion function and bactericidal activity.


Activation of Ras and the mitogen-activated protein kinase pathway promotes protein degradation in muscle cells of Caenorhabditis elegans.
To discover and study intracellular signals that regulate proteolysis in muscle, we have employed transgenic strains of Caenorhabditis elegans that produce a soluble LacZ reporter protein limited to body-wall and vulval muscles. This reporter protein is stable in well-fed wild-type animals, but its degradation is triggered upon a shift to 25 degrees C in a strain carrying a temperature-sensitive activating mutation in the Ras oncogene homologue let-60. These mutants are not physiologically starved, inasmuch as growth rates are normal at 25 degrees C. Ras-induced degradation is not prevented by the presence of cycloheximide added at or before the temperature shift and thus uses preexisting proteolytic systems and signaling components. Furthermore, degradation is triggered when adult animals are shifted to conditions of 25 degrees C, confirming that Ras acutely promotes protein degradation in muscles whose developmental history is normal. Reduction-of-function mutations in the downstream protein kinase Raf (lin-45), MEK (mek-2), or mitogen-activated protein kinase (MAPK) (mpk-1) prevent Ras-induced protein degradation, whereas activated MPK-1 is sufficient to trigger degradation, indicating that this kinase cascade is the principal route by which Ras signaling triggers protein degradation in muscle. This pathway is activated in hypodermal cells by the LET-23 epidermal growth factor receptor homologue, but an activating mutation in let-23 does not promote proteolysis in muscle. Starvation-induced LacZ reporter degradation is unaffected by reduction-of-function mutations in Ras, Raf, MEK, or MAPK, implying that Ras activation and starvation trigger proteolysis by mechanisms that are at least partially independent. This is the first evidence that Ras-Raf-MEK-MAPK signaling activates protein degradation in differentiated muscle.


Electronic structure of alloxan and its dimers: QM/QD simulations and quantum chemical topology analysis.
This study aims to identify the origin of the extra stability of alloxan, a biologically active pyrimidine. To achieve this goal, detailed DFT computations and quantum dynamics simulations have been performed to establish the most stable conformation and the global minimum structure on the alloxan potential energy surface. The effects of the solvent, basis set, and DFT method have been examined to validate the theoretical model adopted throughout the work. Two non-covalent intermolecular dimers of alloxan, the H-bonded and dipolar dimers, have been investigated at the ωB97X-D and M06-2X levels of theory using the triple zeta 6-311++G** to establish their relative stability. Quantum chemical topology features and natural bond orbital analysis (NBO) have been performed to identify and characterize the forces that govern the structures and underlie the extra stability of alloxan.


Is aluminum related bone disease common in hemodialysis units using aluminum based phosphate binders?
The extent of aluminum related bone disease was evaluated in 41 patients on regular maintenance hemodialysis in two dialysis centers in Riyadh, Saudi Arabia. There were 22 males and 19 females aged 20 to 70 years (mean 35.5+11.2 years). Thirty eight of the patients were on aluminum based oral phosphate binders. Investigations performed included serum calcium, magnesium, aluminum and parathormone and radiological investigations including skeletal survey and dual photon absorptiometry. All the patients underwent bone biopsy and the biopsy material was subjected to morphometric studies, including staining for aluminum deposits. The patients were divided into two groups: group 1, (16 patients, 39%) with negative aluminum staining in the bone biopsy tissue, and group 2, (25 patients, 61%) with positive aluminum staining. Bone pain and its distribution as well as fractures were similarly prevalent in both groups. The levels of aluminum in the blood was significantly higher in group 2 (32.9 + 20.2 vs 17.9 + 11.2 ug/L P< 0.05), though it was lower than the lowest accepted toxic level (40 ig/L). There was no significant difference between the two groups in the biochemical or hormonal data, frequency of abnormal radiological signs and pattern of bone histology. Our study indicates that increased aluminum deposition in the bone is prevalent in patients on maintenance hemodialysis using aluminum based phosphate binders, but adynamic bone disease is not prevalent. Further studies may be needed on a larger scale to assess the magnitude of the problem.


A new multi-tracer pellet injection for a simultaneous study of low- and mid/high-Z impurities in high-temperature plasmas.
A new multi-tracer technique in the Tracer-Encapsulated Solid Pellet (TESPEL) method has been developed in order to acquire simultaneously the information about the behaviors of various impurities, i.e., to study concurrently the behaviors of low- and mid/high-Z impurities in magnetically confined high-temperature plasmas. In this new technique, an inorganic compound (for example, lithium titanate, Li2TiO3) is proposed to be used as a tracer embedded in the core of the TESPEL, instead of pure elements. The results of the proof-of-principle experiment clearly demonstrate the applicability of the new multi-tracer technique in the TESPEL method for the simultaneous study of behaviors of low- and mid/high-Z impurities in high-temperature plasmas.


New echocardiographic techniques for evaluating left ventricular myocardial function.
Ultrasound imaging of the heart continues to play an important role in diagnosis and management of patients with cardiovascular diseases. Recent advances in ultrasound technology and introduction of newer imaging modalities have enabled improved assessment of left ventricular myocardial function. Tissue Doppler imaging and 2-dimensional speckle tracking allow more objective quantification of myocardial function in the form of tissue velocities, displacement, strain, and strain rate. Similarly, contrast-enhanced echocardiography and 3-dimensional echocardiography have provided a unique insight into left ventricular form and function that was not possible by unenhanced 2-dimensional echocardiography. In this review, the authors discuss the clinical application of these new imaging techniques in the assessment of left ventricular myocardial function.


Negative regulation contributes to tissue specificity of the immunoglobulin heavy-chain enhancer.
We have identified in and around the immunoglobulin heavy-chain enhancer two apparently distinct negative regulatory elements which repress immunoglobulin H enhancer, simian virus 40 enhancer, and heterologous promoter activity in fibroblasts but not in myeloma cells. We propose that in nonlymphoid cells, negative regulatory elements prevent activation of the immunoglobulin H enhancer by ubiquitous stimulatory trans-acting factors.


Persistence of diminished bone mineral content following renal transplantation in childhood.
Bone mineral content (BMC) was measured in the nondominant arm of 18 children (aged 3 7/12 to 17 1/2 years) for a total 783 months after renal transplantation. Using photon absorptiometry, 89 measurements were made; 17 of the 18 patients had a functioning graft and one patient died. Significant demineralization, a BMC greater than -2 SD below appropriate control volumes, was found in 11 of 18 patients (62%) and 55 of 89 measurements (61%). Bone loss was progressive; among the 16 patients followed for more than 6 months, ten showed a decline of more than 0.5 SD in BMC, five had no change, and only one showed improvement. No relationship was found between BMC and the use of furosemide, type of transplant (15 living, seven cadaver), prior renal disease (six with glomerulonephritis, 11 tubulointerstitial), need for a second graft (five patients), chronic anticonvulsant therapy, or serum calcium and phosphate values. BMC was slightly correlated (P less than .05) with alkaline phosphatase values. BMC was more strongly correlated with serum creatinine (y = -0.48x + 1.25, r = -.042, P less than .001) and prednisone dose (mg/kg/d) (y = -0.65x + 0.481, r = -.543, P less than .001) in an inverse relationship. Patients whose serum creatinine value was less than 1 mg/dL had a BMC of -0.71 +/- 0.34 SD; those with serum creatinine value greater than 2 mg/dL had BMC of -3.32 +/- 0.31 SD, different at P less than .001. Patients receiving daily prednisone therapy had a significantly lower BMC than those receiving alternate-day therapy (-3.11 +/- 1.23 SD v -1.72 +/- 1.29 SD, P less than .005).(ABSTRACT TRUNCATED AT 250 WORDS)


A study of falls among elderly persons in a rural area of Haryana.
Falls are a common, disabling, and frequently fatal health concern among elderly persons. Assessment of the prevalence of falls and associated factors can lead to the identification of corrective measures, which can help in preventing falls and their consequent effects on health and well-being of the elderly. The objective is to determine the prevalence of falls among elderly persons in a rural area and to study the association of falls with sociodemographic variables and selected health conditions. In a community-based, cross-sectional study conducted among 456 elderly persons in a rural area, information regarding sociodemographic details, selected health conditions, and history of falls in the past 12 months was recorded. Univariate analysis followed by stepwise multivariate logistic regression analysis was carried out. The effect of sociodemographic and various health conditions on falls was analyzed using logistic regression analysis. Among the 456 study participants, the prevalence of falls in the past 12 months was 36.6% (95% confidence interval [CI] =32.1-40.0). The prevalence among women was 40.6% (95% CI = 34.5-46.7) and among men was 31.5% (95% CI = 25.0-37.9). Low socioeconomic status, urgency of micturition, knee pain, visual impairment, hearing impairment, functional disability, and depression were significantly associated with falls. Falls are common among elderly persons. Health programs for the elderly must include prevention of falls and rehabilitation of fall-related injuries.


Acute phase cytotoxic T lymphocyte escape is a hallmark of simian immunodeficiency virus infection.
Cytotoxic T-lymphocyte (CTL) responses peak coincident with the decline in acute HIV viremia. Despite two reports of CTL-resistant HIV variants emerging during acute infection, the contribution of acute CTL escape to HIV pathogenesis remains unclear. Difficulties inherent in studying acute HIV infection can be overcome by modeling virus-host interactions in SIV-infected rhesus macaques. We sequenced 21 complete simian immunodeficiency virus (SIV)mac239 genomes at four weeks post-infection to determine the extent of acute CTL escape. Here we show that viruses from 19 of 21 macaques escaped from CTLs during acute infection and that these escape-selecting CTLs were responsive to lower concentrations of peptide than other SIV-specific CTLs. Interestingly, CTLs that require low peptide concentrations for stimulation (high 'functional avidity') are particularly effective at controlling other viral infections. Our results suggest that acute viral escape from CTLs is a hallmark of SIV infection and that CTLs with high functional avidity can rapidly select for escape variants.


Derivative discontinuity and exchange-correlation potential of meta-GGAs in density-functional theory.
We investigate fundamental properties of meta-generalized-gradient approximations (meta-GGAs) to the exchange-correlation energy functional, which have an implicit density dependence via the Kohn-Sham kinetic-energy density. To this purpose, we construct the most simple meta-GGA by expressing the local exchange-correlation energy per particle as a function of a fictitious density, which is obtained by inverting the Thomas-Fermi kinetic-energy functional. This simple functional considerably improves the total energy of atoms as compared to the standard local density approximation. The corresponding exchange-correlation potentials are then determined exactly through a solution of the optimized effective potential equation. These potentials support an additional bound state and exhibit a derivative discontinuity at integer particle numbers. We further demonstrate that through the kinetic-energy density any meta-GGA incorporates a derivative discontinuity. However, we also find that for commonly used meta-GGAs the discontinuity is largely underestimated and in some cases even negative.


Prophylactic Pregabalin to Decrease Pain During Medication Abortion: A Randomized Controlled Trial.
To evaluate whether prophylactic pregabalin reduces pain experienced with medication abortion. We conducted a randomized, double-blind, placebo-controlled trial of women initiating medication abortion with mifepristone and buccal misoprostol up to 70 days of gestation. Participants were randomized to 300 mg oral pregabalin or a placebo immediately before misoprostol. The primary outcome was maximum pain on an 11-point numerical rating scale, reported using real-time electronic surveys over 72 hours. Secondary outcomes included pain at each time point, ibuprofen and narcotic use, side effects, and satisfaction. We estimated that 110 women would be required to have 80% power to detect a difference in pain of 1.3 points. Between June 2015 and October 2016, 241 women were screened and 110 were randomized (56 pregabalin, 54 placebo). Three were lost to follow-up. The primary outcome of mean maximum pain in the pregabalin group was 5.0±2.6 vs 5.5±2.2 in the placebo group (P=.32). Excluding medication taken before the study capsule, ibuprofen was used by 64% (35/55) of the pregabalin group vs 87% (45/52) placebo (P<.01). Narcotics were used by 29% (16/55) of the pregabalin group vs 50% (26/52) placebo (P<.03). More dizziness (P<.001), sleepiness (P<.04), and blurred vision (P<.05) occurred in the pregabalin group. Satisfaction scores for the analgesic regimen were higher in the pregabalin group (very satisfied: 47% vs 22%; P=.006). Compared with placebo, 300 mg pregabalin coadministered with misoprostol during medication abortion did not significantly decrease maximum pain scores. Women who received pregabalin were less likely to require any ibuprofen or narcotic and were more likely to report higher satisfaction with analgesia, despite an increase in dizziness, sleepiness, and blurred vision. ClinicalTrials.gov, NCT02782169.


Endovascular repair of peripheral aneurysms, pseudoaneurysms, and arteriovenous fistulas.
Endovascular repair of peripheral arterial lesions was performed in 10 patients including two iliac aneurysms, two iliac anastomotic pseudoaneurysms, one subclavian pseudoaneurysm, one axillary anastomotic disruption, two prosthetic pseudoaneurysms, and two posttraumatic arteriovenous (AV) fistulas. The indications for repair were aneurysm size in five cases, massive hematoma in one, threatened prosthetic dialysis graft in two, venous hypertension with non-healing ulcer in one, and arm pain in one. Vascular access was obtained through surgical cutdown in all cases, via the femoral artery in five patients, the proximal brachial artery in three and a prosthetic graft in two. Stented prosthetic grafts were used in five cases (1 polyester and polytetrafluoroethylene 4 [PTFE]), and PTFE-covered stents in five cases. Concomitant procedures were done in four patients including two open repairs of a common femoral artery aneurysm, a transluminal dilatation of a proximal aortic anastomotic stenosis, and an iliac artery transluminal angioplasty. Eight of 10 cases were technically successful. Completion arteriography revealed complete exclusion of all lesions, except for one minimal proximal stented graft leak in a pseudoaneurysm, and an incomplete obliteration of an AV fistula. No complications occurred. Operative time ranged from 45 min to 5 hours. Postoperative hospital stay was 1 day in five patients, 2 days in three patients, and 4 days in two patients. Follow-up contrast CT scan, arteriography, or duplex scanning demonstrated complete exclusion of all lesions except an AV fistula, and decrease in size in three aneurysms. The proximal leak initially present in a stented graft resolved. All grafts and covered stents remained patent at 2-19 months of followup. Endovascular exclusion of peripheral arterial aneurysms, pseudoaneurysms, and AV fistulas can be done with a high degree of technical success, low morbidity, and short hospital stay. Short-term follow up is encouraging, however, long term follow up of these procedures is warranted to assess durability of the repair and absence of complications.


Trust, but Verify II: A Practical Guide to Chemogenomics Data Curation.
There is a growing public concern about the lack of reproducibility of experimental data published in peer-reviewed scientific literature. Herein, we review the most recent alerts regarding experimental data quality and discuss initiatives taken thus far to address this problem, especially in the area of chemical genomics. Going beyond just acknowledging the issue, we propose a chemical and biological data curation workflow that relies on existing cheminformatics approaches to flag, and when appropriate, correct possibly erroneous entries in large chemogenomics data sets. We posit that the adherence to the best practices for data curation is important for both experimental scientists who generate primary data and deposit them in chemical genomics databases and computational researchers who rely on these data for model development.


Assessment and characterization of the total geometric uncertainty in Gamma Knife radiosurgery using polymer gels.
This work proposes and implements an experimental methodology, based on polymer gels, for assessing the total geometric uncertainty and characterizing its contributors in Gamma Knife (GK) radiosurgery. A treatment plan consisting of 26, 4-mm GK single shot dose distributions, covering an extended region of the Leksell stereotactic space, was prepared and delivered to a polymer gel filled polymethyl methacrylate (PMMA) head phantom (16 cm diameter) used to accurately reproduce every link in the GK treatment chain. The center of each shot served as a "control point" in the assessment of the GK total geometric uncertainty, which depends on (a) the spatial dose delivery uncertainty of the PERFEXION GK unit used in this work, (b) the spatial distortions inherent in MR images commonly used for target delineation, and (c) the geometric uncertainty contributor associated with the image registration procedure performed by the Leksell GammaPlan (LGP) treatment planning system (TPS), in the case that registration is directly based on the apparent fiducial locations depicted in each MR image by the N-shaped rods on the Leksell localization box. The irradiated phantom was MR imaged at 1.5 T employing a T2-weighted pulse sequence. Four image series were acquired by alternating the frequency encoding axis and reversing the read gradient polarity, thus allowing the characterization of the MR-related spatial distortions. MR spatial distortions stemming from main field (B0) inhomogeneity as well as from susceptibility and chemical shift phenomena (also known as sequence dependent distortions) were found to be of the order of 0.5 mm, while those owing to gradient nonlinearities (also known as sequence independent distortions) were found to increase with distance from the MR scanner isocenter extending up to 0.47 mm at an Euclidean distance of 69.6 mm. Regarding the LGP image registration procedure, the corresponding average contribution to the total geometric uncertainty ranged from 0.34 to 0.80 mm. The average total geometric uncertainty, which also includes the GK spatial dose delivery uncertainty, was found equal to (0.88 ± 0.16), (0.88 ± 0.26), (1.02 ± 0.09), and (1.15 ± 0.24) mm for the MR image series acquired with the read gradient polarity (direction) set toward right, left, posterior, and anterior, respectively. The implemented methodology seems capable of assessing the total geometric uncertainty, as well as of characterizing its contributors, ascribed to the entire GK treatment delivery (i.e., from MR imaging to GK dose delivery) for an extended region of the Leksell stereotactic space. Results obtained indicate that the selection of both the frequency encoding axis and the read gradient polarity during MRI acquisition may affect the magnitude as well as the spatial components of the total geometric uncertainty.


Selection of Effective HTRA3 Activators Using Combinatorial Chemistry.
Herein, we report selection, synthesis, and enzymatic evaluation of a peptidomimetic library able to increase proteolytic activity of HtrA3 (high temperature requirement A) protease. Iterative deconvolution in solution of synthesized modified pentapeptides yielded two potent HtrA3 activators acting in the micromolar range (HCOO-CH2O-C6H4-OCH2-CO-Tyr-Asn-Phe-His-Asn-OH and HCOO-CH2O-C6H4-OCH2-CO-Tyr-Asn-Phe-His-Glu-OH). Both compounds increased proteolysis of an artificial HtrA3 substrate over 40-fold in a selective manner. On the basis of molecular modeling, the selected compounds bind strongly to the PDZ domain.


Mobile medical apps for patient education: a graded review of available dermatology apps.
The utilization of mobile applications (apps) as educational resources for patients highlights the need for an objective method of evaluating the quality of health care-related mobile apps. In this study, a quantified rubric was developed to objectively grade publicly available dermatology mobile apps with the primary focus of patient education. The rubric included 5 criteria thought to be most important in evaluating the adequacy of these apps in relaying health information to patients: educational objectives, content, accuracy, design, and conflict of interest. A 4-point scale was applied to each criterion. The use of this objective rubric could have implications in the evaluation and recommendation of mobile health care apps as a vital educational resource for patients.


[Clinicopathologic characteristics of head and neck carcinoma showing thymus-like element].
Objective: To investigate clinicopathological features of carcinoma showing thymus-like elements (CASTLE) in the head and neck regions. Methods: Clinicopathological data of 7 patients with CASTLE in the head and neck regions were retrospectively reviewed.Immunohistochemical staining and in situ hybridization for EBER were performed. BRAF(V600E) mutation was examined by ARMS method in 6 cases. Results: There were 5 females and 2 males with age between 49 and 78 years (average of 65.6 years). All tumors were solitary nodular lesions with an infiltrative border, including 6 intrathyroid tumors and 1 extrathyroid tumor in the laryngeal pharynx.The tumors were 1.7-4.1 cm in diameter (average of 3.0 cm). Four cases demonstrated lymph node metastasis.All patients were alive without metastasis during follow-up, except one consultation case (with FNA sample) developed recurrence at the primary site. The cases showed different immunoreaction to CD5, diffuse immunoreaction with p63, CK5/6 and CD117, but negative staining for TTF1, TG and calcitonin. One case showed positive immunoreaction with Synin less than 30% tumor cells. The Ki-67 labeling index was between 3% and 90%. No BRAF(V600E) mutation and EB virus infection were detected. Conclusions: Extrathyroid CASTLE involving laryngeal pharynx shows the similar morphological and immunohistochemical features with intrathyroid CASTLE.Immunohistochemical markers of CD5 and CD117 are helpful in the diagnosis. Ki-67 labeling index can be high in CASTLE, especially in lymphoepithelioma type. CD5-negative CASTLE may have neuroendocrine differentiation. BRAF(V600E) mutation and EB virus may not be involved in the carcinogenesis of CASTLE.


Methodological and Cellular Factors Affecting the Magnitude of Breast Cancer and Normal Cell Radiosensitization Using Gold Nanoparticles.
Breast cancer (BC) is the most common malignant tumor in women, which most often originates from the epithelial tissue of the breast gland. One of the most recommended kinds of treatment is radiotherapy (RT), but irradiation (IR) can affect not only the cancer tumor but also the healthy tissue around it. Au nanoparticles (AuNPs) were proposed as a radiosensitizing agent for RT which would allow for lower radiation doses, reducing the negative radiation effects on healthy tissues. The main objective of the study is to assess the dependence on the radiosensitivity of BC (MDA-MB-231) and normal mammary gland epithelial cells (MCF12A) to ionizing radiation, caused by functionalized AuNPs under diverse conditions. The viability, uptake, reactive oxygen species induction, and mitochondrial membrane potential in cells were analyzed applying a time and concentration-dependent manner. After different incubation times with AuNPs, cells were exposed to 2 Gy. The determination of radiation effect in combination with AuNPs was investigated using the clonogenic assay, p53, and γH2AX level, as well as, Annexin V staining. Our results highlighted the strong need for assessing the experimental conditions' optimization before the AuNPs will be implemented with IR. Moreover, results indicated that AuNPs did not act universally in cells. AuNPs could be a promising tool as a radiotherapy sensitizing agent, but it should be specified and deeply investigated under what conditions it will be applied taking into consideration not only AuNPs modifications but also the model and experimental conditions.


Integrating Next-Generation Genomic Sequencing and Mass Spectrometry To Estimate Allele-Specific Protein Abundance in Human Brain.
Gene expression contributes to phenotypic traits and human disease. To date, comparatively less is known about regulators of protein abundance, which is also under genetic control and likely influences clinical phenotypes. However, identifying and quantifying allele-specific protein abundance by bottom-up proteomics is challenging since single nucleotide variants (SNVs) that alter protein sequence are not considered in standard human protein databases. To address this, we developed the GenPro software and used it to create personalized protein databases (PPDs) to identify single amino acid variants (SAAVs) at the protein level from whole exome sequencing. In silico assessment of PPDs generated by GenPro revealed only a 1% increase in tryptic search space compared to a direct translation of all human transcripts and an equivalent search space compared to the UniProtKB reference database. To identify a large unbiased number of SAAV peptides, we performed high-resolution mass spectrometry-based proteomics for two human post-mortem brain samples and searched the collected MS/MS spectra against their respective PPD. We found an average of ∼117 000 unique peptides mapping to ∼9300 protein groups for each sample, and of these, 977 were unique variant peptides. We found that over 400 reference and SAAV peptide pairs were, on average, equally abundant in human brain by label-free ion intensity measurements and confirmed the absolute levels of three reference and SAAV peptide pairs using heavy labeled peptides standards coupled with parallel reaction monitoring (PRM). Our results highlight the utility of integrating genomic and proteomic sequencing data to identify sample-specific SAAV peptides and support the hypothesis that most alleles are equally expressed in human brain.


Tejocote Root's Role in Symptomatic Mobitz Type 1 Heart Block: A Compelling Case Report.
The clinical presentation and diagnosis of Tejocote root toxicity causing Mobitz Type 1 remains a scarcely clinical phenomenon, often resulting in delayed diagnosis and treatment. This case report highlights a 30-year-old female presenting with a constellation of symptoms, including fatigue, dizziness, chest pressure, myalgias, nausea, vomiting, and peripheral tingling. Significantly, the patient had been using Tejocote root as an over-the-counter laxative acquired from Mexico. Laboratory findings revealed detectable Digoxin levels in her bloodstream, while an electrocardiogram (EKG) indicated sinus bradycardia with Mobitz Type 1 heart block. The patient was treated with a single dose of atropine 0.5 mg IV push. A repeat EKG before discharge showed resolution of the Mobitz type 1. This case underscores the potential cardiovascular repercussions of Tejocote root consumption and emphasizes the importance of heightened clinical awareness, especially in regions where such herbal supplement usage is prevalent.


Synthesis and photochemical behavior of phosphorus dendrimers containing azobenzene units within the branches and/or on the surface.
We describe the synthesis of three series of phosphorus-containing dendrimers having azobenzene derivatives specifically placed at some generations in the interior and/or on the surface. The largest compound obtained possesses 48 azobenzene groups on the surface. Irradiation at 350 nm induces isomerization of the azobenzene groups from the E form to the Z form, whatever their location. The thermal back-isomerization to the E form in the dark at room temperature was observed in all cases. The kinetics of this Z-->E back-isomerization was studied in several cases; the rate is not dependent on the number of azobenzene units or of the generation, when the azobenzene groups are linked to the surface of the dendrimer. A different behavior was observed when the azobenzene groups were located within the framework of the dendrimer.


Geographic and temporal validity of prediction models: different approaches were useful to examine model performance.
Validation of clinical prediction models traditionally refers to the assessment of model performance in new patients. We studied different approaches to geographic and temporal validation in the setting of multicenter data from two time periods. We illustrated different analytic methods for validation using a sample of 14,857 patients hospitalized with heart failure at 90 hospitals in two distinct time periods. Bootstrap resampling was used to assess internal validity. Meta-analytic methods were used to assess geographic transportability. Each hospital was used once as a validation sample, with the remaining hospitals used for model derivation. Hospital-specific estimates of discrimination (c-statistic) and calibration (calibration intercepts and slopes) were pooled using random-effects meta-analysis methods. I2 statistics and prediction interval width quantified geographic transportability. Temporal transportability was assessed using patients from the earlier period for model derivation and patients from the later period for model validation. Estimates of reproducibility, pooled hospital-specific performance, and temporal transportability were on average very similar, with c-statistics of 0.75. Between-hospital variation was moderate according to I2 statistics and prediction intervals for c-statistics. This study illustrates how performance of prediction models can be assessed in settings with multicenter data at different time periods.


Designer drugs that are potent inhibitors of CYP2D6.
Some abused drugs are substrates of CYP2D6 (e.g., paramethoxyamphetamine, methylenedioxy-methamphetamine). CYP2D6 inhibition by concurrently used drugs of abuse could potentiate such drugs and increase acute toxicity. Ten designer drugs were tested as inhibitors of CYP2D6. Only 1-methyl-4-phenyl-4-propionoxypiperidine (MPPP) and 1-[2-phenylethyl]-4-phenyl-4-acetoxypiperidine (PEPAP) interacted significantly (Ki 1.6 microM and 0.3 microM, respectively). Both are synthetic analogues of meperidine sold as "synthetic heroin." No CYP2D6-mediated metabolites were detected for either compound. Concurrent oral use of CYP2D6 substrates with MPPP and PEPAP may represent a kinetic drug interaction risk, but this risk must be confirmed clinically.


The burden of bacterial antimicrobial resistance in Croatia in 2019: a country-level systematic analysis.
To deliver the most wide-ranging set of antimicrobial resistance (AMR) burden estimates for Croatia to date. A complex modeling approach with five broad modeling components was used to estimate the disease burden for 12 main infectious syndromes and one residual group, 23 pathogenic bacteria, and 88 bug-drug combinations. This was represented by two relevant counterfactual scenarios: deaths/disability-adjusted life years (DALYs) that are attributable to AMR considering a situation where drug-resistant infections are substituted with sensitive ones, and deaths/DALYs associated with AMR considering a scenario where people with drug-resistant infections would instead present without any infection. The 95% uncertainty intervals (UI) were based on 1000 posterior draws in each modeling step, reported at the 2.5% and 97.5% of the draws' distribution, while out-of-sample predictive validation was pursued for all the models. The total burden associated with AMR in Croatia was 2546 (95% UI 1558-3803) deaths and 46958 (28,033-71,628) DALYs, while the attributable burden was 614 (365-943) deaths and 11321 (6,544-17,809) DALYs. The highest number of deaths was established for bloodstream infections, followed by peritoneal and intra-abdominal infections and infections of the urinary tract. Five leading pathogenic bacterial agents were responsible for 1808 deaths associated with resistance: Escherichia coli, Staphylococcus aureus, Acinetobacter baumannii, Klebsiella pneumoniae, and Pseudomonas aeruginosa (ordered by the number of deaths). Trimethoprim/sulfamethoxazole-resistant E coli and methicillin-resistant S. aureus were dominant pathogen-drug combinations in regard to mortality associated with and attributable to AMR, respectively. We showed that AMR represented a substantial public health concern in Croatia, which reflects global trends; hence, our detailed country-level findings may fast-track the implementation of multipronged strategies tailored in accordance with leading pathogens and pathogen-drug combinations.


Polarizability of red blood cells with an anisotropic membrane.
We predict the complex polarizability of a realistic model of a red blood cell (RBC), with an inhomogeneous dispersive and anisotropic membrane. In this model, the frequency-dependent complex electrical parameters of the individual cell layers are described by the Debye equation while the dielectric anisotropy of the cell membrane is taken into account by the different permittivities along directions normal and tangential to the membrane surface. The realistic shape of the RBC is described in terms of the Jacobi elliptic functions. To calculate the polarizability, we evoke the effective dipole moment method to determine the cell internal electric field distribution, employing an adaptive finite-element numerical approach. We have furthermore investigated the influence of the anisotropic membrane and dispersive electrical parameters of each individual cell layer on the total complex polarizability. Our findings suggest that the individual layer contribution depends on two factors: the volume of the layer and the associated induced electric field, which in turn is influenced by other layers of the cell. These results further show that the average polarizability spectra of the cell are significantly impacted by the anisotropy and associated dispersion of the cellular compartments.


Salmeterol and fluticasone propionate (50/250 microg) administered via combination Diskus inhaler: as effective as when given via separate Diskus inhalers.
To compare the efficacy and safety of a new combination Diskus inhaler containing both salmeterol 50 mg and fluticasone propionate 250 mg (Seretide) with the two drugs delivered via separate Diskus inhalers. A multicentre, double-blind, double-dummy study. Three hundred and seventy-one symptomatic asthma patients (age range 13 to 75 years, mean 42 years) receiving inhaled corticosteroids were randomly assigned to two treatment groups: 28 weeks' treatment with either salmeterol/fluticasone propionate (50/250 microg bid) via a single Diskus inhaler (combination) and placebo bid via another Diskus inhaler, or salmeterol 50 microg bid via one Diskus inhaler and fluticasone propionate 250 microg bid via another (concurrent). Morning peak expiratory flow rate (PEFR) and symptoms were measured for the first 12 weeks and safety data were collected throughout the study. Over weeks 1 to 12, adjusted mean improvements in morning PEFR were 43 and 36 L/min for combination and concurrent therapies, respectively. The difference between the two treatment arms was 6 L/min (90% CI -13 to 0 L/min; P=0.114), which was within the predefined criteria for clinical equivalence. Adjusted mean improvements in forced expiratory volume in 1 s from baseline for week 28 were also similar between the two therapies. Thirty-five per cent of patients receiving combination inhaler and 31% of those receiving concurrent therapy had a mean daytime symptom score of zero over weeks 1 to 12 compared with 1% and 2%, respectively, at baseline. There was no difference in the incidence of adverse events between the two treatment arms. Mean serum cortisol levels were similar, and no differences in frequency of abnormal results were noted between the two groups. This study shows that the combination of salmeterol and fluticasone propionate in a single inhaler is as efficacious in achieving asthma control and as well tolerated over a 28-week period as the two drugs administered individually.


Immunomodulating effects and hypersensitivity reactions caused by Northeast Indian black fly salivary gland extract.
The genus Simulium, black fly (Diptera), comprises >1800 species worldwide, of which 67 species under six subgenera have been well studied in India. While at the extreme, black fly bites can cause onchocerciasis or river blindness, the majority of clinical observations indicate mainly severe pain and continuous itching at bite sites. This investigation experimentally observed that black fly salivary extract (BFSE) causes unique biologic effects including immunomodulation, anti-coagulation, and hypersensitivity reactions in Wistar rats. Salivary glands from black flies were isolated, extracted in saline, and then ≈800 ng extract (BFSE) subcutaneously injected into rats. To evaluate potential immunosuppressive activity of the BFSE, serum levels of interleukins [IL]-6 and -10 and tumor necrosis factor [TNF]-α were assayed. To assess the BFSE impact on coagulation, activated partial thromboplastin time (aPTT), prothrombin time (PT), and bleeding time, as well as generation of coagulation activated factors I, IX, and X were analyzed. Anaphylaxis induction was monitored via electrocardiogram (ECG) and measures of blood pressure and rectal temperature. The data showed that BFSE treatment resulted in a significantly prolonged aPTT, PT, and bleeding time and reversibly inhibited generation of coagulation activated factors I, IX, and X. The extract also led to a prolonged (up to 48 h) suppression of serum IL-6, IL-10, and TNFα production. While these results suggest that BFSE possesses anti-thrombotic, anti-coagulant, and immunomodulatory activities, importantly, they also indicate that the extract has a capacity to induce anaphylaxis and acute cardiotoxicity.


Neurofibromatosis Type 1-Associated Optic Pathway Gliomas: Current Challenges and Future Prospects.
Optic pathway glioma (OPG) occurs in as many as one-fifth of individuals with the neurofibromatosis type 1 (NF1) cancer predisposition syndrome. Generally considered low-grade and slow growing, many children with NF1-OPGs remain asymptomatic. However, due to their location within the optic pathway, ~20-30% of those harboring NF1-OPGs will experience symptoms, including progressive vision loss, proptosis, diplopia, and precocious puberty. While treatment with conventional chemotherapy is largely effective at attenuating tumor growth, it is not clear whether there is much long-term recovery of visual function. Additionally, because these tumors predominantly affect young children, there are unique challenges to NF1-OPG diagnosis, monitoring, and longitudinal management. Over the past two decades, the employment of authenticated genetically engineered Nf1-OPG mouse models have provided key insights into the function of the NF1 protein, neurofibromin, as well as the molecular and cellular pathways that contribute to optic gliomagenesis. Findings from these studies have resulted in the identification of new molecular targets whose inhibition blocks murine Nf1-OPG growth in preclinical studies. Some of these promising compounds have now entered into early clinical trials. Future research focused on defining the determinants that underlie optic glioma initiation, expansion, and tumor-induced optic nerve injury will pave the way to personalized risk assessment strategies, improved tumor monitoring, and optimized treatment plans for children with NF1-OPG.


[The findings of computed tomography (CT) and magnetic resonance imaging (MRI) in pulmonary arterial hypertension].
Technical advances of multidetector-row computed tomography(MDCT) and magnetic resonance imaging(MRI) made these modalities more important in the evaluation and for differential diagnosis of pulmonary arterial hypertension (PAH). The advantages of CT and MRI are noninvasive examination, wide field of view, excellent reproducibility, high spatial resolution and 3-dimensional (3-D) reconstruction images. Morphological changes of the PAH, which are right ventricular hypertrophy and dilatation of main and central pulmonary artery(PA), right ventricle, right atria, superior and inferior vena cava, and coronary sinus, are depicted well by these modalities. The 3-D CT and MR angiography can depict peripheral and central PA, and pulmonary veins, which are important information of the cause of PAH. Myocardial changes of PAH can be detected by gadolinium delayed enhancement of MRI. CT and MRI are promising method to diagnose and manage the PAH in future.


Factor interactions at simian virus 40 GC-box promoter elements in intact nuclei.
Primer extension footprinting was used to probe late simian virus 40 regulatory elements in intact infected cell nuclei. Specific protection was observed over the viral "GC-box" transcription elements. The participation of the bound templates in gene activation is addressed by quantitation that shows that their abundance greatly exceeds that of transcription complexes but is comparable to that of open chromatin.


The 3.5 ångström X-ray structure of the human connexin26 gap junction channel is unlikely that of a fully open channel.
The permeability of gap junction channels to metabolites, and not simply to small inorganic ions, is likely to play an important role in development, physiology as well as in etiology of several diseases. Here, we combined dual patch clamp and fluorescence imaging techniques with molecular dynamics (MD) simulations to investigate the permeation of calcein, a relatively large fluorescent tracer (MW 622 Da) through homomeric gap junction channels formed by wild type human connexin26 (hCx26wt) protomers. Our experimental data indicate that the unitary flux of calcein driven by a 125 μM concentration difference is Jpore = 226 molecule/s per channel. In the light of Eyring transition state theory adapted for the liquid phase, this value corresponds to an energy barrier of ~20 kBT (where kB is the Boltzmann constant and T is absolute temperature). The barrier predicted by our MD simulations, based on the 3.5 Å X-ray structural model of the hCx26wt gap junction channel, is ~45 kBT. The main contributions to the energetics of calcein permeation originated from the interaction between the permeating molecule and the charged aminoacids lining the channel pore. Assigning a fake zero total charge to the calcein molecule yielded a value for the barrier height compatible with the experimental data. These results can be accounted for by two different (although not mutually exclusive) hypotheses: (1) the X-ray model of the hCx26wt gap junction channel is not representative of a fully open state; (2) post translational modifications affecting the hCx26wt protein in our expression system differed from the modifications undergone by the proteins in the conditions used to obtain the crystal structure. Hypothesis (1) is compatible with data indicating that, only 10% or less of the channels forming a gap junction plaque are in the open state, and therefore the averaging procedure intrinsic in the generation of the crystal structure data more closely reflects that of a closed channel. Hypothesis (2) is compatible with recent mass spectrometry data and implies that the charge of several amino acid side chains may have been altered, thus modifying substantially the permeation properties of the channels in living cells.


Utilization of health care resources by HIV-infected white, African-American, and Hispanic men in the era before highly active antiretroviral therapy.
In the highly active antiretroviral therapy (HAART) era, U.S. African-American and Hispanic patients with HIV use HAART less, but emergency and inpatient services more, than white patients. We evaluated whether these patterns existed in the pre-HAART era. Data from prospective cohort studies of 462 male Veterans Affairs patients and 1,309 male patients from the AIDS Costs and Services Utilization Survey were combined. Resource utilization of white, African-American, and Hispanic men was compared. Compared to whites, African Americans were more likely to visit the emergency department and less likely to have mental health, home health, and dental visits; had fewer outpatient and substance abuse treatment visits; and had more inpatient nights. Hispanics were less likely to have mental health and home health visits, and had more inpatient nights. Whites used prescription drugs more than African Americans or Hispanics, but antiretrovirals were equally used. Lower access to HAART for African-American and Hispanic patients is a new phenomenon, not a continuation of pre-HAART patterns, while the undesirable patterns of emergency and outpatient provider resource utilization in the HAART era are a continuation of pre-HAART patterns. Undesirable resource utilization patterns by African-American and Hispanic populations need urgent attention.


Toward high throughput 3D virtual screening using spherical harmonic surface representations.
Searching chemical databases for possible drug leads is often one of the main activities conducted during the early stages of a drug development project. This article shows that spherical harmonic molecular shape representations provide a powerful way to search and cluster small-molecule databases rapidly and accurately. Our clustering results show that chemically meaningful clusters may be obtained using only low order spherical harmonic expansions. Our database search results show that using low order spherical harmonic shape-based correlation techniques could provide a practical and efficient way to search very large 3D molecular databases, hence leading to a useful new approach for high throughput 3D virtual screening. The approach described is currently being extended to allow the rapid search and comparison of arbitrary combinations of molecular surface properties.


Comparison of rhinovirus A infection in human primary epithelial and HeLa cells.
HeLa cells are used to study the life cycles of many different viruses, including the human rhinoviruses (HRV) in the family Picornaviridae. Although the natural targets of HRV are human bronchial epithelial cells (hBE), it is generally more difficult to obtain and maintain the relevant primary cell cultures, relative to HeLa cells. Given that the HRV are now identified as a major cause of human asthma exacerbations, it becomes important to document how much of the virus biology learned from HeLa cells is common also to natural primary cells. When compared directly in matched infections using A01a virus, the kinetics of RNA replication, the synthesis and processing of viral proteins and the general subcellular localization of key non-structural proteins were resembled in hBE and HeLa cells. Viral-induced shutoff of host cell processes (e.g. nucleo-cytoplasmic trafficking) was also comparable.


Comparison of denaturation of tryptophan synthase alpha-subunits from Escherichia coli, Salmonella typhimurium, and an interspecies hybrid.
Guanidine hydrochloride-induced denaturation and thermal denaturation of three kinds of tryptophan synthase alpha subunit have been compared by circular dichroism measurements. The three alpha subunits are from Escherichia coli, Salmonella typhimurium, and an interspecies hybrid in which the C-terminal domain comes from E. coli (alpha-2 domain) and the N-terminal domain comes from S. typhimurium (alpha-1 domain). Analysis of denaturation by guanidine hydrochloride at 25 degrees C showed that the alpha-2 domain of S. typhimurium was more stable than the alpha-2 domain of E. coli, but the alpha-1 domain of S. typhimurium was less stable than the alpha-1 domain of the E. coli protein; overall, the hybrid protein was slightly less stable than the two original proteins. It is concluded that the stability to guanidine hydrochloride denaturation of each of the domains of the interspecies hybrid is similar to the stability of the domain of the species from which it originated. The E. coli protein was more stable to thermal denaturation than the other proteins near the denaturation temperature, but the order of their thermal stability was reversed at 25 degrees C and coincided with that obtained from guanidine hydrochloride-induced denaturation.


How common are side effects of spinal manipulation and can these side effects be predicted?
Little scientific support is available concerning usual and unusual reactions after spinal manipulation although such reactions are very common in clinical practice. Fifty-nine manipulative therapists were requested to enroll 15 consecutive patients attending for their first visit to receive spinal manipulation. These patients were asked to complete a questionnaire after this first visit that asked for possible risk factors for spinal manipulation and asked about any side effects after the manipulation. The participating practitioners were asked to note medical diagnosis, manipulated spinal region, number of treated areas and type of additional treatment. Four hundred and sixty five valuable responses were analysed. Two hundred and eighty three patients (60.9%) reported at least one post-manipulative reaction. The most common were headache (19.8%), stiffness (19.5%), local discomfort (15.2%), radiating discomfort (12.1%) and fatigue (12.1%). Most of these reactions began within 4 h and generally disappeared within the next 24 h. Women were more likely to report adverse effects than men (P < 0.001). Multivariate analysis showed that upper cervical manipulation (OR: 3.20; 95%CI: 1.89-7.77), use of medication (OR: 2.20; 95%CI: 1.31-3.69), gender (OR: 1.66; 95%CI: 1.01-2.75) and age (OR: 1.02; 95%CI: 1.00-1.05) were independent predictors of headache after spinal manipulation. The results of this study indicate that reactions to spinal manipulation may be relatively common but are benign in nature and of short duration. Although it is difficult to label side effects as a risk, it is important to differentiate those patients who are susceptible to side effects in order to inform them correctly.


Genetic regulation of the immune response to hepatitis B surface antigen (HBsAg). I. H-2 restriction of the murine humoral immune response to the a and d determinants of HBsAg.
In hepatitis B virus (HBV) infection correlation of disease activity with major histocompatibility complex (MHC) antigens has been suggested. However, no data are available regarding the nature and regulation of the immune response to the surface viral protein (HBsAg). This consideration and the current emphasis on production of an HBV vaccine led us to investigate the humoral immune response to HBsAg in inbred strains of mice to assess genetic factors that may regulate the humoral anti-HBs response. Studies with H-2 congenic and noncongenic inbred strains revealed the T-dependent humoral immune responses to the group-specific a and subtype-specific d determinants of HBsAg are regulated by gene(s) mapping to the murine MHC. The H-2q haplotype conferred high responsiveness, the H-2s,f haplotypes conferred low to nonresponsiveness, and the H-2a,b,d,k haplotypes were intermediate. Studies of H-2 recombinant strains allowed tentative mapping of regulation of the anti-a response and possibly the anti-d response to the K/I-A, I-B region of the MHC. The H-2-restricted regulation of the humoral immune response to HBsAg was circumvented by immunization with 10-fold higher HBsAg doses. These findings demonstrate a clear linkage between the murine MHC and the regulation of the immune response to specific determinants on HBsAg.


Vascular endothelial growth factor levels in ascites between chemonaive and chemotreated patients.
Vascular endothelial growth factor (VEGF) levels in malignant ascites have high diagnostic value for their discrimination from ascites of non-malignant origin. However, there have been no reports on the comparison of VEGF levels between malignant ascites of chemonaive and chemotreated patients. VEGF levels were measured in 44 ascites patients (cirrhosis ascites, 10; chemonaive patients, 21; chemotreated patients, 13) and compared to the level of carcinoembryonic antigen (CEA) and carbohydrate antigen 19-9 (CA 19-9). The diagnostic parameters of sensitivity, specificity, and correlation among 3 markers were evaluated. VEGF levels in malignant ascites of chemonaive and chemotreated patients were significantly higher than those in cirrhotic ascites (p<0.05). VEGF levels in ascites of chemonaive patients were significantly higher than those in chemotreated patients (p<0.05). A cutoff value of 10.4 pg/mL was calculated using receiver operating characteristic curves (ROCs) for VEGF in chemotreated and chemonaive patients, which gave sensitivities of 75.0% and 53.8% and specificities of 69.6% and 47.1%, respectively. Positive correlations were observed between VEGF and CEA (r=0.353, p<0.05) as well as between VEGF and CA19-9 (r=0.367, p<0.05) in ascites. VEGF levels could be a useful tumor marker for malignant ascites, but its value should carefully be interpreted because of lesser reliability in chemotreated ones.


Anxiolytic effects of a novel herbal treatment in mice models of anxiety.
Anxiety and stress disorders are currently among the ten most important public health concerns, and in recent years, have reached epidemic proportions. The current success rate of treatments for anxiety disorders is not high, reaching 50% at most. These treatments are also associated with a wide variety of side effects. The aim of the present study was to investigate the anxiolytic properties of a novel herbal treatment produced in our laboratory compared to a conventional treatment for anxiety disorders, namely SSRIs. Anxiety-like behavior was evaluated in adult mice exposed to stress during childhood following 1, 2 and 3 weeks of treatment with the novel herbal treatment or escitalopram, using the novel open field and the elevated plus maze paradigms. The behavioral evaluation in these mice was followed by a biochemical assessment of their brain hippocampal BDNF levels and blood corticosterone levels. The study showed that (1) the novel herbal treatment reduced anxiety-like behaviors in both behavioral tests. Interestingly, this reduction was observed only following a 3-week treatment; (2) following the novel treatment, corticosterone levels in the plasma of treated mice were reduced and this reduction was similar to the one observed following escitalopram treatment; and (3) BDNF levels in the hippocampus of mice treated both with the novel herbal treatment and escitalopram were increased. These behavioral and biological findings indicate that our novel herbal compound has the potential of being highly efficacious in treating anxiety disorders.


Nasal Columellar Reconstruction With Reverse Lateral Nasal Artery Pedicled Nasolabial Island Flap.
Nasal columellar is a complex area to reconstruct due to its unique esthetic and functional characteristics. Malignancy, trauma, infection, and esthetic surgery complications can cause columellar defect. Among many surgical plans, nasolabial island flap is most commonly used. However, flap congestions occasionally occur which inevitably lead to leech therapy. The authors introduce reverse lateral nasal artery pedicled nasolabial island flap, which has lower risk of postoperative complications.


Towards Communicative Justice in Health.
This article approaches care from a different angle by looking ethnographically at how it is shaped by structural differences in the power to control the circulation of knowledge. I focus on an investigation conducted by people classified as "indigenous", of an epidemic that killed 38 children and young adults in a Venezuelan rainforest. I trace how health/communicative inequities structured clinical interactions, documents, epidemiological investigations, news stories, and dialogues with healers, thwarting the identification of the epidemic, clinically identified as rabies. Although the Bolivarian socialist government provided access to care, professionals denigrated parents' contributions to care and communication and reduced complex, unequal relations between languages to practical problems of translation. Pointing to parallels with US social movements, I suggest that responding to demands for communicative justice in health requires seeing how health inequities are entangled with health/communicative inequities. The typographical slash points to importance of challenging the subdisciplinary boundary-work that relegates their study to non-overlapping conversations in medical and linguistic anthropology.