Interaction of herpes simplex virus type 2 (HSV-2) glycoprotein D with the host cell surface is sufficient to induce Chlamydia trachomatis persistence.
When presented with certain unfavourable environmental conditions, Chlamydia trachomatis reticulate bodies (RBs) enter into a viable, yet non-cultivable state called persistence. Previously, we established an in vitro C. trachomatis and herpes simplex virus type 2 (HSV-2) co-infection model. These data indicate that (i) viral co-infection stimulates chlamydial persistence, (ii) productive HSV replication is not required for persistence induction, and (iii) HSV-induced persistence is not mediated by any currently characterized anti-chlamydial pathway or persistence inducer. In this study we demonstrated that chlamydial infectivity, though initially suppressed, recovered within 44 h of co-infection with UV-inactivated HSV-2, demonstrating that HSV-induced persistence is reversible. Co-incubation of chemically fixed, HSV-2-infected inducer cells with viable, C. trachomatis-infected responder cells both suppressed production of infectious chlamydial progeny and stimulated formation of swollen, aberrantly shaped RBs. In addition, pre-incubation of viral particles with viral glycoprotein D (gD)-specific neutralizing antibody prevented co-infection-induced persistence. Finally, exposure of C. trachomatis-infected cells to a soluble, recombinant HSV-2 gD : Fc fusion protein decreased production of infectious EBs to a degree similar to that observed in co-infected cultures. Thus, we conclude that interaction of HSV gD with the host cell surface is sufficient to trigger a novel host anti-chlamydial response that restricts chlamydial development.


Permeation of roxarsone and its metabolites increases caco-2 cell proliferation.
The benzenearsonate, Roxarsone, has been used since 1944 as an antimicrobial, growth-promoting poultry feed additive. USGS and EPA report that Roxarsone (4-hydroxy-3-nitrobenzenearsonate) and metabolites, including AHBA (3-amino-4-hydroxybenzenearsonate), contaminate waterways at greater than 1100 tons annually. To assess human impact of these organic arsenic water contaminants, it was important to study their potential absorption. The human adenocarcinoma cell line, Caco-2, is a model for intestinal absorption. We found proliferative effects on Caco-2 cells at micromolar levels of these compounds, as monitored by [3H]-thymidine incorporation into DNA. Flow cytometry cell cycle analysis confirmed accumulation in S phase from 21% (control) to 36% (24 hour exposure to 10 μM AHBA). Confluent Caco-2 cells grown on collagen-coated Transwell plates were dosed on the apical side. After exposure, media from apical and basolateral sides were collected separately. Following removal of FBS by 30K centrifugal filtration, the benzenearsonates in the collected media were analyzed by HPLC. Analyses were at wavelengths in the ultraviolet/visible range where the absorbance values were linear with respect to concentration. Concentrations were calculated by comparison with analytically-prepared commercial standards. Results from cells dosed at 10 μM for 24 hours with AHBA, Roxarsone, or Acetarsone indicated 6% - 29% permeation occurring from apical to basolateral side, modeling absorption across intestinal epithelium to the circulatory system. Benzenearsonate feed additives are frequently applied in combination with antibiotics, raising additional health concerns. We conclude that micromolar levels of these benzenearsonates are adequate to stimulate Caco-2 cell proliferation.


Antenna Modification Leads to Enhanced Nitrogenase Activity in a High Light-Tolerant Cyanobacterium.
Biological nitrogen fixation is an energy-intensive process that contributes significantly toward supporting life on this planet. Among nitrogen-fixing organisms, cyanobacteria remain unrivaled in their ability to fuel the energetically expensive nitrogenase reaction with photosynthetically harnessed solar energy. In heterocystous cyanobacteria, light-driven, photosystem I (PSI)-mediated ATP synthesis plays a key role in propelling the nitrogenase reaction. Efficient light transfer to the photosystems relies on phycobilisomes (PBS), the major antenna protein complexes. PBS undergo degradation as a natural response to nitrogen starvation. Upon nitrogen availability, these proteins are resynthesized back to normal levels in vegetative cells, but their occurrence and function in heterocysts remain inconclusive. Anabaena 33047 is a heterocystous cyanobacterium that thrives under high light, harbors larger amounts of PBS in its heterocysts, and fixes nitrogen at higher rates compared to other heterocystous cyanobacteria. To assess the relationship between PBS in heterocysts and nitrogenase function, we engineered a strain that retains large amounts of the antenna proteins in its heterocysts. Intriguingly, under high light intensities, the engineered strain exhibited unusually high rates of nitrogenase activity compared to the wild type. Spectroscopic analysis revealed altered PSI kinetics in the mutant with increased cyclic electron flow around PSI, a route that contributes to ATP generation and nitrogenase activity in heterocysts. Retaining higher levels of PBS in heterocysts appears to be an effective strategy to enhance nitrogenase function in cyanobacteria that are equipped with the machinery to operate under high light intensities. IMPORTANCE The function of phycobilisomes, the large antenna protein complexes in heterocysts has long been debated. This study provides direct evidence of the involvement of these proteins in supporting nitrogenase activity in Anabaena 33047, a heterocystous cyanobacterium that has an affinity for high light intensities. This strain was previously known to be recalcitrant to genetic manipulation and, hence, despite its many appealing traits, remained largely unexplored. We developed a genetic modification system for this strain and generated a ΔnblA mutant that exhibited resistance to phycobilisome degradation upon nitrogen starvation. Physiological characterization of the strain indicated that PBS degradation is not essential for acclimation to nitrogen deficiency and retention of PBS is advantageous for nitrogenase function.


GFP as a Genetic Marker Scorable Throughout the Life Cycle of Transgenic Zebra Fish.
A fish expression vector, FRM, was constructed by fusing the carp beta-actin promoter and first intron to the ocean pout antifreeze protein terminator and putative boundary element. Mutant forms of the green fluorescent protein (GFP) were engineered into this vector, and the resultant series of vectors, FRMwg, FRM3wg (green GFP), and FRM2bl (blue GFP), were used to make transgenic zebra fish. After microinjection of either supercoiled or linearized DNA into one-celled eggs, GFP-expressing cells could be monitored by fluorescence microscopy commencing with the midblastula transition and continuing through embryogenesis. From adult fish, which retained scorable GFP either as patches or as a uniform fluorescence, 11 green and 1 blue GFP-expressing lines of zebra fish have been established. Expression of GFP was nearly ubiquitous and similar among all of these lines. Embryonic expression could be scored at 15 to 30 hours postfertilization and was seen throughout the embryo with the exceptions of the yolk, red blood cells, and in some lines, portions of the head. Adult expression was in a majority of tissues (e.g., muscle, brain, intestine, and heart, but not red blood cells). The notable difference between lines was that fluorescent eggs were scorable in seven of the lines. Adult homozygotes from a different subset of eight lines could be selected by the relative intensity of the GFP marking when compared with that in sibling heterozygotes. All 12 lines contain apparent single locus, multicopy, tandem integrations (1.5-100 copies per cell) of the transgenic DNA. The fish expression vector FRM could be used to drive nearly ubiquitous and strong expression of gene products other than GFP. The GFP expression vectors, FRMwg, FRM2wg, FRM3wg, and FRM2bl, may be useful for optimization of transgenesis and as a comarker. GFP-expressing zebra fish lines could facilitate experimental analysis of chimerism and in vivo gene targeting.


The D1/D5 Dopamine Partial Agonist PF-06412562 in Advanced-Stage Parkinson's Disease: A Feasibility Study.
Current drug treatments have little efficacy in advanced-to-end-stage Parkinson's disease (advPD), yet there are no reports of interventional trials in advPD. D1 dopamine agonists have the potential to provide benefit. To determine the feasibility and safety of the selective D1/D5 dopamine partial agonist PF 06412562 in advPD. A two-week, randomized, double blind, crossover phase Ib study in advPD patients compared standard-of-care (SoC) carbidopa/levodopa with PF 06412562. Each week, there was a Day 1 baseline evaluation with overnight levodopa washout, then treatment on Days 2 and 3 with either SoC or PF-06412562 (split dose 25 + 20 mg), followed by discharge on Day 4. Primary endpoints were safety and tolerability. Secondary endpoints were global clinical impression of change (GCI-C) rated by clinicians and caregivers. Eight advPD patients and their caregivers consented to participate and six were randomized (average disease duration: 22 y). None withdrew voluntarily. One participant with baseline Day 1 dehydration, pre-renal kidney injury, and autonomic dysfunction experienced symptomatic and serious hypotension after receiving PF-06412562 in Week 1 and was discontinued from the study. All other adverse events were rated mild (PF-06412562: n = 1, SoC: n = 0), moderate (PF-06412562: n = 1, SoC: n = 1), or severe but non-serious (PF-06412562: n = 3, SoC: n = 2). No clinically meaningful laboratory changes were observed. Among the five participants who completed the study, GCI-C favored PF-06412562 in two per clinicians' and four participants per caregivers' rating. PF-06412562 was tolerated in advPD patients. This study provides the feasibility for future safety and efficacy studies in this population with unmet needs.


Protein synthesis inhibitors stimulate MondoA transcriptional activity by driving an accumulation of glucose 6-phosphate.
Protein synthesis is regulated by the availability of amino acids, the engagement of growth factor signaling pathways, and adenosine triphosphate (ATP) levels sufficient to support translation. Crosstalk between these inputs is extensive, yet other regulatory mechanisms remain to be characterized. For example, the translation initiation inhibitor rocaglamide A (RocA) induces thioredoxin-interacting protein (TXNIP). TXNIP is a negative regulator of glucose uptake; thus, its induction by RocA links translation to the availability of glucose. MondoA is the principal regulator of glucose-induced transcription, and its activity is triggered by the glycolytic intermediate, glucose 6-phosphate (G6P). MondoA responds to G6P generated by cytoplasmic glucose and mitochondrial ATP (mtATP), suggesting a critical role in the cellular response to these energy sources. TXNIP expression is entirely dependent on MondoA; therefore, we investigated how protein synthesis inhibitors impact its transcriptional activity. We investigated how translation regulates MondoA activity using cell line models and loss-of-function approaches. We examined how protein synthesis inhibitors effect gene expression and metabolism using RNA-sequencing and metabolomics, respectively. The biological impact of RocA was evaluated using cell lines and patient-derived xenograft organoid (PDxO) models. We discovered that multiple protein synthesis inhibitors, including RocA, increase TXNIP expression in a manner that depends on MondoA, a functional electron transport chain and mtATP synthesis. Furthermore, RocA and cycloheximide increase mtATP and G6P levels, respectively, and TXNIP induction depends on interactions between the voltage-dependent anion channel (VDAC) and hexokinase (HK), which generates G6P. RocA treatment impacts the regulation of ~ 1200 genes, and ~ 250 of those genes are MondoA-dependent. RocA treatment is cytotoxic to triple negative breast cancer (TNBC) cell lines and shows preferential cytotoxicity against estrogen receptor negative (ER-) PDxO breast cancer models. Finally, RocA-driven cytotoxicity is partially dependent on MondoA or TXNIP. Our data suggest that protein synthesis inhibitors rewire metabolism, resulting in an increase in mtATP and G6P, the latter driving MondoA-dependent transcriptional activity. Further, MondoA is a critical component of the cellular transcriptional response to RocA. Our functional assays suggest that RocA or similar translation inhibitors may show efficacy against ER- breast tumors and that the levels of MondoA and TXNIP should be considered when exploring these potential treatment options.


Ovine herpesvirus 2 replicates initially in the lung of experimentally infected sheep.
Ovine herpesvirus 2 (OvHV-2), a rhadinovirus in the subfamily Gammaherpesvirinae, is the causative agent of sheep-associated malignant catarrhal fever (SA-MCF), a frequently fatal lymphoproliferative disease primarily of ruminants worldwide. Inability to propagate the virus in vitro has made it difficult to study OvHV-2 replication. Aerosol inoculation of sheep with OvHV-2 from nasal secretions collected from naturally infected sheep during shedding episodes results in infection of naive sheep, providing an excellent system to study OvHV-2 initial replication in the natural host. In this study, we showed that OvHV-2 delivered through the nasal route by nebulization resulted in infection in all lambs, but no infection was established in any lambs after intravenous or intraperitoneal injection. In nebulized lambs, while it was not detected initially in any other tissues, OvHV-2 DNA became detectable in the lung at 3 days post-infection (p.i.), increased to about 900 copies per 50 ng DNA at 5 days p.i., reached peak levels ( approximately 7500 copies) at 7 days p.i., and then declined to an average of 800 copies at 9 days p.i. Transcripts of OvHV-2 open reading frame 25 (coding for the capsid protein), an indicator of virus replication, were only detected in lung tissues, and were positively correlated with OvHV-2 DNA levels in the lungs. In addition, selected immune response genes were also highly expressed in the lung at 5 and 7 days p.i. The data indicate that lung is the primary replication site for OvHV-2 during initial infection in sheep and suggest that viral replication is promptly controlled by a host defence mechanism.


In situ bioremediation of perchlorate in vadose zone soil using gaseous electron donors: microcosm treatability study.
A series of microcosm experiments were performed to determine the effectiveness of various gaseous electron donors (including hydrogen, 1-hexene, ethyl acetate, and liquefied petroleum gas [LPG]) for supporting biological perchlorate reduction under different electron donor concentrations and soil moistures. Under high soil moisture (16% w/w) conditions, complete or partial perchlorate degradation was achieved with all of the tested electron donors, except for ethyl acetate. Hydrogen was the most promising of the tested electron donors, achieving complete perchlorate degradation with first-order rate constants ranging from 0.13 to 0.20 day(-1) and reducing concentrations to non-detectable levels within 35 to 42 days. The LPG and 1-hexene each promoted partial perchlorate reduction, with average first-order rate constants of 0.05 and 0.11 day(-1), respectively. Although significant perchlorate reduction was observed with as little as 13% moisture, the moisture content for complete perchlorate degradation in this particular soil was determined to be 17%.


Synthesis and release of platelet-activating factor is inhibited by plasma alpha 1-proteinase inhibitor or alpha 1-antichymotrypsin and is stimulated by proteinases.
TNF and IL-1 stimulate the synthesis and release of platelet-activating factor (PAF) by neutrophils and vascular endothelial cells. Serum inhibits PAF production even after inactivation of an acetylhydrolase that degrades PAF. Human plasma was fractionated by gel filtration chromatography, and two inhibitory fractions were detected, one containing PAF-acetylhydrolase activity and the other alpha 1-proteinase inhibitor. Low concentrations of this antiproteinase and of human plasma alpha 1-antichymotrypsin inhibited TNF-induced PAF synthesis in neutrophils, macrophages, and vascular endothelial cells. Both antiproteinases also inhibited PAF production stimulated by phagocytosis in macrophages and induced with IL-1 in neutrophils or with TNF in vascular endothelial cells. These results suggest that a proteinase activated on the plasma membrane or secreted by these cells is involved in promoting PAF synthesis. Indeed, addition of elastase to macrophages, neutrophils, and endothelial cells stimulated synthesis and release of PAF much faster than TNF. A similar stimulation was observed in incubations with cathepsin G. To identify a proteinase activated in TNF-treated cells, neutrophils and endothelial cells were incubated with specific chloromethyl ketone inhibitors of elastase and cathepsin G. Synthesis of PAF was significantly inhibited by low concentrations of the cathepsin G inhibitor. The finding that antiproteinases are inhibitory at concentrations 100-fold lower than those present in plasma raises questions as to the ability of TNF and IL-1 to stimulate neutrophils in circulation or endothelial cells to synthesize PAF. We propose that PAF production is limited to zones of close contact between cells, which exclude antiproteinases.


Th2 LCR is essential for regulation of Th2 cytokine genes and for pathogenesis of allergic asthma.
Previous studies have shown that Th2 cytokine genes on mouse chromosome 11 are coordinately regulated by the Th2 locus control region (LCR). To examine the in vivo function of Th2 LCR, we generated CD4-specific Th2 LCR-deficient (cLCR KO) mice using Cre-LoxP recombination. The number of CD4 T cells in the cLCR KO mouse was comparable to that in wild-type mice. The expression of Th2 cytokines was dramatically reduced in in vitro-stimulated naïve CD4 T cells. Deletion of the LCR led to a loss of general histone H3 acetylation and histone H3-K4 methylation, and demethylation of DNA in the Th2 cytokine locus. Upon ovalbumin challenge in the mouse model of allergic asthma, cLCR KO mice exhibited marked reduction in the recruitment of eosinophils and lymphocytes in the bronchoalveolar lavage fluid, serum IgE level, lung airway inflammation, mucus production in the airway walls, and airway hyperresponsiveness. These results directly demonstrate that the Th2 LCR is critically important in the regulation of Th2 cytokine genes, in chromatin remodeling of the Th2 cytokine locus, and in the pathogenesis of allergic asthma.


Three-dimensional structures of RNA obtained by means of knowledge-based interaction potentials.
We derive a set of effective potentials describing the interaction between pairs of nucleotides that belong to an RNA molecule. Such interaction potentials are then used as the main constituents of a simplified simulation model, which is tested in the description of small secondary structure motifs. Our simulated RNA hairpins are consistent with the experimental structures obtained by NMR.


How do patients with systemic sclerosis experience currently provided healthcare and how should we measure its quality?
To gain insight into SSc patients' perspective on quality of care and to survey their preferred quality indicators. An online questionnaire about healthcare setting, perceived quality of care (CQ index) and quality indicators, was sent to 2093 patients from 13 Dutch hospitals. Six hundred and fifty patients (mean age 59 years, 75% women, 32% limited cutaneous SSc, 20% diffuse cutaneous SSc) completed the questionnaire. Mean time to diagnosis was 4.3 years (s.d. 6.9) and was longer in women compared with men (4.8 (s.d. 7.3) vs 2.5 (s.d. 5.0) years). Treatment took place in a SSc expert centre for 58%, regional centre for 29% or in both for 39% of patients. Thirteen percent of patients was not aware of whether their hospital was specialized in SSc. The perceived quality of care was rated with a mean score of 3.2 (s.d. 0.5) (range 1.0-4.0). There were no relevant differences between expert and regional centres. The three prioritized process indicators were: good patient-physician interaction (80%), structural multidisciplinary collaboration (46%) and receiving treatment according to SSc guidelines (44%). Absence of disease progression (66%), organ involvement (33%) and digital ulcers (27%) were the three highest rated outcome indicators. The perceived quality of care evaluated in our study was fair to good. No differences between expert and regional centres were observed. Our prioritized process and outcome indicators can be added to indicators suggested by SSc experts in earlier studies and can be used to evaluate the quality of care in SSc.


Pathogenesis of viral hemorrhagic fevers: Rift Valley fever and Lassa fever contrasted.
Although many viral infections have on occasion been associated with hemorrhagic complications, infection with any of several RNA viruses regularly results in vascular involvement and the syndrome called viral hemorrhagic fever (VHF). In spite of clinically useful similarities among various VHFs, there are significant differences in their pathogenesis and clinical evolution; these are often related to characteristics of their viral taxon. Infection with Rift Valley fever (RVF) virus, a phlebovirus, appears to be regulated by interferon and terminated by neutralizing antibody. In contrast, Lassa fever (LF) virus, an arenavirus, is resistant to interferon, and LF is terminated by cellular immune effector mechanisms. The lytic virus-cell interaction typical of RVF virus suggests its major effects occur by direct, virus-induced cellular necrosis, particularly in the liver. In the primate RVF model, disseminated intravascular coagulation (DIC) may be important. LF virus--characteristically noncytopathic--may exert its effects through induction of mediator secretion from infected macrophages. DIC does not appear to be a central pathogenetic mechanism in LF. Pichinde virus, which is not pathogenic for humans, provides an alternate model for study of LF. Infected guinea pigs do not show histologic lesions that could explain their body wasting, cardiovascular deterioration, and pulmonary edema. In the heart, for example, loss of tissue mass, protein, and contractile function proceed without direct viral involvement or myocarditis. Sulfidopeptide leukotrienes have been implicated as one relevant soluble mediator participating in the disease state.


Stereotactic LINAC-Radiosurgery for Glomus Jugulare Tumors: A Long-Term Follow-Up of 27 Patients.
The optimal treatment of glomus jugulare tumors (GJTs) remains controversial. Due to the critical location, microsurgery still provides high treatment-related morbidity and a decreased quality of life. Thus, we performed stereotactical radiosurgery (SRS) for the treatment of GJTs and evaluated the long-term outcome. Between 1991 and 2011, 32 patients with GJTs underwent SRS using a linear accelerator (LINAC) either as primary or salvage therapy. Twenty-seven patients (median age 59.9 years, range 28.7-79.9 years) with a follow-up greater than five years (median 11 years, range 5.3-22.1 years) were selected for retrospective analysis. The median therapeutic single dose applied to the tumor surface was 15 Gy (range 11-20 Gy) and the median tumor volume was 9.5 ml (range 2.8-51 ml). Following LINAC-SRS, 10 of 27 patients showed a significant improvement of their previous neurological complaints, whereas 12 patients remained unchanged. Five patients died during follow-up due to old age or other, not treatment-related reasons. MR-imaging showed a partial remission in 12 and a stable disease in 15 patients. No tumor progression was observed. The actuarial overall survival rates after five, ten and 20 years were 100%, 95.2% and 79.4%, respectively. Stereotactic LINAC-Radiosurgery can achieve an excellent long-term tumor control beside a low rate of morbidity in the treatment of GJTs. It should be considered as an alternative therapy regime to surgical resection or fractionated external beam radiation either as primary, adjuvant or salvage therapy.


Management of upper extremity dysfunction in people with Parkinson disease and Huntington disease: facilitating outcomes across the disease lifespan.
Parkinson Disease (PD) and Huntington Disease (HD) are degenerative neurological diseases, which can result in impairments and activity limitations affecting the upper extremities from early in the disease process. The progressive nature of these diseases poses unique challenges for therapists aiming to effectively maximize physical functioning and minimize participation restrictions in these patient groups. Research is underway in both diseases to develop effective disease-modifying agents and pharmacological interventions, as well as mobility-focused rehabilitation protocols. Rehabilitation, and in particular task-specific interventions, has the potential to influence the upper extremity functional abilities of patients with these degenerative conditions. However to date, investigations of interventions specifically addressing upper extremity function have been limited in both PD, and in particular HD. In this paper, we provide an update of the known pathological features of PD and HD as they relate to upper extremity function. We further review the available literature on the use of outcome measures, and the clinical management of upper extremity function in both conditions. Due to the currently limited evidence base in both diseases, we recommend utilization of a clinical management framework specific for degenerative conditions that can serve as a guideline for disease management.


TLR4 Asp299Gly polymorphism may be protective against chronic periodontitis.
Periodontitis results from interplay between genetic and environmental factors. Single nucleotide polymorphisms (SNPs) in the coding region of the toll-like receptor 4 gene (TLR4) may be associated with periodontitis, although previous studies have been inconclusive. Moreover, the interaction between environmental factors, such as cigarette smoking (a major risk factor for periodontitis), and Porphyromonas gingivalis (a major periodontal pathogen) with the TLR4 coding region Asp299Gly SNP (rs4986790; a SNP associated with lipopolysaccharide-mediated inflammatory responses in periodontitis), have been largely ignored in previous reports. Therefore, the objective of this study was to examine the association between TLR4 Asp299Gly (rs4986790) with alveolar bone height loss (ABHL) and periodontitis, accounting for interactions between this SNP with smoking and P. gingivalis prevalence. The CD14/-260 SNP (rs2569190) served as a control, as a recent meta-analysis suggested no relationship between this SNP and periodontitis. This multicenter study included 617 participants who had rheumatoid arthritis or osteoarthritis. This report presents a secondary outcome from the primary case-control study examining the relationship of periodontitis with established rheumatoid arthritis. The Centers for Disease Control/American Academy of Periodontology case definitions of periodontitis were used for this analysis. Participants received a full-mouth clinical periodontal examination and panoramic radiograph. Percentage ABHL was measured on posterior teeth. The TLR4 Asp299Gly and CD14/-260 SNPs were selected a priori and genotypes were determined using the ImmunoChip array (Illumina(®) ). Minor allele frequencies and associations with periodontitis and ABHL did not differ according to rheumatoid arthritis vs. osteoarthritis status; therefore, data from these two groups were pooled. The presence of P. gingivalis was detected in subgingival plaque by PCR. Multivariate ordinal logistic regression examined associations between the SNPs and periodontitis or ABHL. SNP interactions with smoking and P. gingivalis were analyzed. A significant, negative interaction was observed between the TLR4 SNP and the presence of P. gingivalis (p = 0.045) with respect to periodontitis. The TLR4 minor variant was also associated with less ABHL: 16.8% of individuals with low ABHL, 9.0% with moderate ABHL and 11.2% with high ABHL had the minor allele [p = 0.029; odds ratio = 0.58 (95% confidence interval: 0.36-0.95)]. The interaction between the TLR4 SNP and smoking was not significant with respect to periodontitis or ABHL. The CD14 SNP was not associated with periodontitis or ABHL. The TLR4 Asp299Gly SNP significantly interacted with P. gingivalis in conferring a decreased risk of periodontitis and may be protective against ABHL, a feature of periodontitis. Agents blocking TLR4 signaling, a strategy currently under investigation for the treatment of other inflammatory conditions, may warrant investigation in the context of periodontitis related to the presence of P. gingivalis.


Experimental optimization of protein refolding with a genetic algorithm.
Refolding of proteins from solubilized inclusion bodies still represents a major challenge for many recombinantly expressed proteins and often constitutes a major bottleneck. As in vitro refolding is a complex reaction with a variety of critical parameters, suitable refolding conditions are typically derived empirically in extensive screening experiments. Here, we introduce a new strategy that combines screening and optimization of refolding yields with a genetic algorithm (GA). The experimental setup was designed to achieve a robust and universal method that should allow optimizing the folding of a variety of proteins with the same routine procedure guided by the GA. In the screen, we incorporated a large number of common refolding additives and conditions. Using this design, the refolding of four structurally and functionally different model proteins was optimized experimentally, achieving 74-100% refolding yield for all of them. Interestingly, our results show that this new strategy provides optimum conditions not only for refolding but also for the activity of the native enzyme. It is designed to be generally applicable and seems to be eligible for all enzymes.


[Sublingual gland amyloidosis causing obstructive sleep apnea hypopnea syndrome: a case report and review of the literatures].
To improve the understanding of sublingual gland amyloidosis causing obstructive sleep apnea hypopnea syndrome(OSAHS). A case of sublingual gland amyloidosis causing OSAHS diagnosed in april 2012 was reported and the related literatures were reviewed. The literature review was carried out respectively with "amyloidosis, sublingual gland, obstructive sleep apnea hypopnea syndrome", as the search terms in Wanfang Data and PubMed by November 2012. A case of 74 year-old male patient was admitted to the hospital because of snoring for 5 years, sleep apnea for 1 year and arousal for 1 month. After admission, polysomnography showed severe OSAHS, physical examination showed redundant the sublingual gland. Enhanced CT scanning showed soft tissue masses at the sublingual gland. Abdominal B ultrasonic and CT also showed a spaces-occupying lesion in the left retroperitoneal. B-guided core needle biopsy was performed in the left retroperitoneal. Pathology report showed amyloidosis. Subsequently, sublingual gland mass resection was performed. Pathology report after operation showed amyloid deposits staining with Congo red, which gives it a characteristic green birefringence in polarised light. Accordingly, it was diagnosed as sublingual gland amyloidosis. The symptoms of snore and sleep apnea were disappeared after operation.So far, there was no local recurrence with 10 months follow-up. A total of 3 literatures were received in Wanfang Data, including 2 of macroglossia amyloidosis causeing OSAHS case report and one of retrospective study. There were no reports about sublingual gland amyloidosis in Wanfang Data. A total of 5 literatures were received in Pubmed, including 2 of sublingual gland amyloidosis case report, 2 of macroglossia amyloidosis causeing OSAHS case report and one of retrospective study. However, there were no reports about sublingual gland amyloidosis causing OSAHS. Amyloidosis rarely occurred in the sublingual gland and is easy to be misdiagnosed and missed diagnosed, which can causing severe OSAHS. To make a definite diagnosis, histopathology and staining with Congo red are needed and a characteristic green birefringence in polarised light is a reliable marker for diagnosis. After sublingual gland mass resection, the patient had good prognosis.


Electron microscopic studies of mitosis in amebae. I. Amoeba proteus.
Individual organisms of Amoeba proteus have been fixed in buffered osmium tetroxide in either 0.9 per cent NaCl or 0.01 per cent CaCl(2), sectioned, and studied in the electron microscope in interphase and in several stages of mitosis. The helices typical of interphase nuclei do not coexist with condensed chromatin and thus either represent a DNA configuration unique to interphase or are not DNA at all. The membranes of the complex nuclear envelope are present in all stages observed but are discontinuous in metaphase. The inner, thick, honeycomb layer of the nuclear envelope disappears during prophase, reappearing after telophase when nuclear reconstruction is in progress. Nucleoli decrease in size and number during prophase and re-form during telophase in association with the chromatin network. In the early reconstruction nucleus, the nucleolar material forms into thin, sheet-like configurations which are closely associated with small amounts of chromatin and are closely applied to the inner, partially formed layer of the nuclear envelope. It is proposed that nucleolar material is implicated in the formation of the inner layer of the envelope and that there is a configuration of nucleolar material peculiar to this time. The plasmalemma is partially denuded of its fringe-like material during division.


Late Pleistocene age and archaeological context for the hominin calvaria from GvJm-22 (Lukenya Hill, Kenya).
Kenya National Museums Lukenya Hill Hominid 1 (KNM-LH 1) is a Homo sapiens partial calvaria from site GvJm-22 at Lukenya Hill, Kenya, associated with Later Stone Age (LSA) archaeological deposits. KNM-LH 1 is securely dated to the Late Pleistocene, and samples a time and region important for understanding the origins of modern human diversity. A revised chronology based on 26 accelerator mass spectrometry radiocarbon dates on ostrich eggshells indicates an age range of 23,576-22,887 y B.P. for KNM-LH 1, confirming prior attribution to the Last Glacial Maximum. Additional dates extend the maximum age for archaeological deposits at GvJm-22 to >46,000 y B.P. (>46 kya). These dates are consistent with new analyses identifying both Middle Stone Age and LSA lithic technologies at the site, making GvJm-22 a rare eastern African record of major human behavioral shifts during the Late Pleistocene. Comparative morphometric analyses of the KNM-LH 1 cranium document the temporal and spatial complexity of early modern human morphological variability. Features of cranial shape distinguish KNM-LH 1 and other Middle and Late Pleistocene African fossils from crania of recent Africans and samples from Holocene LSA and European Upper Paleolithic sites.


Effect of the urgency and landing zone on rates of in-hospital death, stroke, and paraplegia after thoracic endovascular aortic repair in Japan.
The present study used data from the Japanese Committee for Stentgraft Management's national registry, which contains unique surgical data, including surgical timing, anatomic factors, and pathologic factors, to determine the generalized community experience with thoracic endovascular abdominal aortic repair (TEVAR). The medical background and short-term outcomes were reviewed for patients who had undergone TEVAR for a thoracic aortic aneurysm (TAA; 14,235 cases) or aortic dissection (AD; 990 type A and 4259 type B) from 2008 to 2015. TEVAR for AD was separated from that for TAAs; only the background and short-term outcomes were evaluated. The technical outcomes of TEVAR for TAA were also evaluated. All the cases were categorized as follows: elective, urgent (within 24 hours after admission), or emergent (immediately after admission). The outcomes included in-hospital mortality and persistent stroke and paraplegia diagnosed at discharge. The number of debranching bypasses, proximal landing zone (0, 1, 2, ≥3), and zone length were included in the logistic regression analysis. The mortality, stroke, and paraplegia rates in the TAA and AD groups were 4.4%, 4.6%, and 3.7% and 4.0%, 2.9%, and 2.8%, respectively. After analyzing the TAA cohort, we found that urgent and emergent cases were associated with all adverse outcomes. The rate of paraplegia increased drastically in the patients with stent graft coverage that extended for six or more zones. Massive atheroma was associated with stroke and paraplegia. The cumulative survival rate of the TAA group was stratified by the urgency (ie, elective, urgent, emergent; P < .001). We found that the more proximal (0, 1, and 2) the landing zone, the greater the risk of stroke. Likewise, the longer (six or more zones) the coverage, the greater the risk of paraplegia. Urgency was strongly associated with mortality, stroke, and paraplegia, and the classification of urgent and emergent, according to the surgical timing after admission, successfully stratified the population in the long-term overall survival analysis. A proximal landing zone involving the aortic arch and debranching bypasses were associated with the occurrence of stroke, and the length of stent graft coverage for six or more zones was associated with paraplegia. Identifying these risk factors will help operators of TEVAR develop appropriate operative strategies to mitigate patient risk.


Thyrotropin receptor-adenylate cyclase function in human thyroid neoplasms.
The action of thyrotropin (TSH) on plasma membranes was studied to elucidate the mechanism of hormonal regulation of malignant versus normal human thyroid tissue. Thyroid plasma membranes of six specimens of papillary or follicular carcinoma and six of adenoma, as well as adjacent normal tissue obtained from these patients, were evaluated with respect to binding of 125I-labeled TSH and stimulation of adenylate cyclase. Scatchard analysis of TSH binding revealed the presence of two species of binding sites in normal thyroid of different affinities and capacities. In 11 of 12 tumors studied, the high-affinity binding site remained intact; however, the total number of low-affinity sites was markedly lower than normal tissue. Other parameters of binding were not altered in neoplastic thyroid. In each of these tissues, the hormone responsiveness and kinetics of adenylate cyclase activation were essentially identical to those observed in normal tissue, although basal activity was typically greater in the neoplasm. One carcinoma was totally deficient in both 125I-labeled TSH binding and TSH-stimulatable adenylate cyclase, although basal activity was detected. Furthermore, adenylate cyclase of this specimen was not activated by prostaglandin, in contrast to normal thyroid and other thyroid tumors. These results suggest that: (a) clinical behavior of thyroid carcinomas may not be reflected by TSH receptor-adenylate cyclase function; (b) lack of clinical response as manifest by tumor regression cannot be ascribed to the absence of functional TSH receptors or adenylate cyclase; and (c) decreased low-affinity binding present in tumors is not correlated with altered hormone responsiveness of adenylate cyclase but may reflect more general cancer-induced changes in membrane structure or composition.


Anode Reaction Control for a Single-Compartment Electrochemical CO2 Reduction Reactor with a Surface-Activated Diamond Cathode.
The electrochemical reaction of carbon dioxide (CO2) in aqueous electrolyte solutions is attracting increasing attention for sustainable chemical production. Boron-doped diamond (BDD) electrodes have been previously shown to be very effective for the stable electrochemical production of formic acid from CO2. Typically, the electrochemical production of formic acid by CO2 reduction (CO2R) reaction is performed with a dual-compartment flow reactor equipped with a membrane separator. The problems caused by the membrane separator, such as scaling-up, complicated operational control and materials costs can be solved using a membrane free single-compartment reactor. Here we demonstrate anode reaction control for a single-compartment CO2R flow reactor using a surface-activated BDD cathode and achieve a Faradaic efficiency for formic acid production of over 70%.


[Clinico-economical aspects of cardioversion of paroxysmal atrial fibrillation at phehospital stage and during hospitalization].
We carried out clinico-economical analysis of 2 tactics of rhythm restoration in patients with paroxysmal atrial fibrillation (AF) lasting less than 48 hours: cardioversion at prehospital stage with intravenous procainamide and inhospital cardioversion with any method. This retrospective study was based on the data from department of urgent aid of an outpatient clinic. The results showed that within 48 hours inhospital was a was more effective, safe, and more economically profitable compared with administration of procainamide at prehospital stage. Intravenous procainamide resulted in effective cardioversion in 70.6% of patients. It was associated with arterial hypotension and proarrhythmogenic action in 14,7% of cases. Patients with effective cardioversion with procainamide had lesser mean values of left ventricular anterior-posterior dimension (echocardiography) and shorter duration of arrhythmia.


Breast implants and cancer risk in Denmark.
Although millions of women worldwide have received breast implants for cosmetic or medical reasons, possible late effects (in particular cancer) have not been well studied. To provide quantitative information on cancer occurrence among women undergoing breast implant surgery, 1,135 women treated for cosmetic reasons in Denmark were evaluated. Patients were identified using the nationwide Hospital Discharge Registry with linkage to the nationwide Danish Cancer Registry to determine subsequent cancer incidence. The average age of the women at implant surgery was 31 years, and the average follow-up was 8.4 years, up to a maximum of 17 years. Overall, 27 cancers developed after implant surgery compared with 24.7 expected based on incidence rates from the general population (standardized incidence ratio [SIR] = 1.1; 95% CI: 0.7-1.6). Eight breast cancers were observed vs. 7.8 expected (SIR = 1.0; 95% CI: 0.4-2.0). No evidence was found to link breast implants with increased cancer risk in the decade after surgery. While the results are encouraging, longer follow-up into later life will be necessary to assess fully any possible adverse effects.


Feasibility of melatonin for prevention of delirium in critically ill patients: a protocol for a multicentre, randomised, placebo-controlled study.
Delirium is highly prevalent in the intensive care unit (ICU) and is associated with adverse clinical outcomes. At this time, there is no drug that effectively prevents delirium in critically ill patients. Alterations in melatonin secretion and metabolism may contribute to the development of delirium. Administration of exogenous melatonin has been shown to prevent delirium in non-critically ill surgical and medical patients. This trial will demonstrate the feasibility of a planned multicentre, randomised controlled trial to test the hypothesis that melatonin can prevent delirium in critically ill patients compared with placebo. This feasibility trial is a randomised, 3-arm, placebo-controlled study of melatonin (2 vs 0.5 mg vs placebo, administered for a maximum of 14 days) for the prevention of delirium in critically ill patients. A total of 69 patients aged 18 years and older with an expected ICU length of stay >48 hours will be recruited from 3 Canadian ICUs. The primary outcome is protocol adherence (ie, overall proportion of study drug doses administered in the prescribed administration window). Secondary outcomes include pharmacokinetic parameters, incidence, time to onset, duration of delirium, number of delirium-free days, adverse events, self-reported sleep quality, rest-activity cycles measured by wrist actigraphy, duration of mechanical ventilation, ICU length of stay and mortality. Data will be analysed using an intention-to-treat approach. The study has been approved by Health Canada and the research ethics board of each study site. Trial results will be presented at international conferences and published in a peer-reviewed journal. NCT02615340: Pre-results.


Correction of extrahepatic portal vein thrombosis by the mesenteric to left portal vein bypass.
The goal of this study was to determine the effectiveness of mesenteric vein to left portal vein bypass operation (MLPVB) in correcting extrahepatic portal vein thrombosis (EHPVT) in children. The treatment of idiopathic EHPVT has been primarily palliative, whereas MLPVB restores hepatic portal flow in patients with EHPVT. Thirty-four children with symptomatic EHPVT underwent surgery with intent to perform MLPVB and were followed for up to 7 years. MLPVB was successful in 31 patients (91%), all of whom maintain patent vein grafts and have symptomatic relief of EHPVT in follow-up. All patients had complete relief from gastrointestinal bleeding. Patients with hypersplenism had significant increases in platelet and leukocyte counts and reduction in spleen size. Superior mesenteric vein flow increased from 119 +/- 66 mL/min before bypass to 447 +/- 225 mL/min (P < 0.0001) after surgery. Postoperative blood flow in the bypass graft expressed as a fraction of calculated ideal portal flow for size correlated inversely with age (P < 0.001). Left-portal vein diameter increased from 2.6 +/- 1.6 mm to 7.3 +/- 2.4 mm 2 years after surgery (P < 0.002). Liver volume increased from 703 +/- 349 cm3 to 799 +/- 351 cm3 1 week after surgery (P < 0.001). Prothrombin time improved to normal in all patients 1 year after surgery. MLPVB provides excellent relief of symptoms in children with idiopathic EHPVT and results in liver growth and normalization of coagulation parameters. This surgery is corrective and should be done at as early an age as possible.


[Gamma-scintigraphy in assessment cardioresynchronizing therapy at chronic heart failure].
The purpose of the work was to study influence of cardioresynchronizing therapy (CRT) on systolic and diastolic myocardial function with the aid of radionuclide methods. 19 patients (9 women) at the age from 21 to 72 years (average age 55.4 +/- 8.3 years) with NYHA class III - IV cardiac insufficiency, dilated cardiomyopathy (n = 14) and ischemic cardiomyopathy (n = 5) were included in the study. Radionuclide angiopulmonography and balanced radionuclide ventriculography were performed by standard method before CRT, in 20 - 30 days and in 12 - 18 months. In 20 - 30 days after CRT significant increase in stroke volume and stroke index, ejection fraction and part of circular fibers contraction was detected, which confirmed improve in systolic heart function. In 12 - 18 months significant increase in maximal filling rate index, filling fraction of 1/3 and 2/3 of diastole was noted, which reflected improve of left ventricle diastolic function. Recovery of myocardial contraction synchronism leaded to rehabilitation of normal pulmonary microcirculation. Radionuclide examination methods as high-informative modes of combine assessment of heart contractile function and pulmonary circulation in CRT dynamics are described. CRT, improving systolic and diastolic myocardial functions, has positive influence on pulmonary hemodynamics in patients with chronic heart failure.


Effect of overlaying rubber on fully slatted concrete floors on hoof health and lying postures in finishing dairy-origin bulls offered two contrasting diets.
Fully slatted concrete floors are labour-efficient, cost-effective and thus common in beef cattle housing. However, the welfare of cattle accommodated on them has been questioned. The objective of this study was to evaluate the effect of floor and diet on hoof health and lying behaviours of housed dairy-origin bulls, from a mean age of 8 months to slaughter at 15.5 months old. Forty-eight bulls, which had a mean initial live weight of 212 (SD = 23.7) kg, were allocated to one of four treatments, which consisted of two floors and two diets arranged in a 2 × 2 factorial design. The floors evaluated were a fully slatted concrete floor and a fully slatted concrete floor overlaid with rubber, while the diets offered were either a high concentrate diet or a grass-silage-based diet supplemented with concentrates. The mean total duration of the study was 216 days. Floor had no significant effect on claw measurements measured on day 62 or 139. However, bulls accommodated on slats overlaid with rubber had a tendency to have a higher front toe length measured pre-slaughter than those accommodated on concrete slats (P = 0.063). Floor had no significant effect on the net growth of toes or heels during the duration of the study. The number of bruises (P < 0.01) and the bruising score (P < 0.05) were significantly higher on day 62 in bulls accommodated on fully slatted concrete floors than on concrete slats overlaid with rubber, but there was no significant effect of floor on these parameters on day 139 or at the measurement taken pre-slaughter. There was a tendency for bulls accommodated on concrete slats to have a higher probability of having sole bruising at the end of the experiment than those accommodated on slats overlaid with rubber (P = 0.052). Diet had no significant effect on toe length or heel height, number of bruises, or overall bruising score at any time point of the study. There was little evidence in the current study to suggest that bulls lying on fully slatted concrete floors could not express lying postures similar to those on concrete slats overlaid with rubber.


Functionalizing micro-3D-printed protein hydrogels for cell adhesion and patterning.
The extracellular matrix has been shown to profoundly influence both cell morphology and numerous cellular processes - including adhesion, differentiation, and alignment - through a range of chemical, mechanical, and topographical features. In these studies, we investigate a versatile platform for functionalizing micro-3D-printed (μ-3DP) protein hydrogels via multiphoton excitation of benzophenone-biotin, a photoactivatable ligand capable of reacting with the hydrogel matrix, which is subsequently linked to a biotinylated cell-adhesive peptide through a NeutrAvidin® bridge. This functionalization strategy is potentially applicable to a broad range of hydrogel platforms, enabling biomolecules to be precisely patterned at specified locations within 3D materials. As proof-of-concept of this strategy's utility, we demonstrate that chemical modifications can be made to μ-3DP protein hydrogels that enable Schwann cells to be patterned without altering the mechanical or topographical properties of the hydrogel to an extent that influences SC cell adhesion. The ability to independently control potential cellular cues within in vitro cellular microenvironments is essential to investigating decoupled effects of biomaterial properties on cell-matrix interactions. In addition, we demonstrate feasibility for generating arbitrary immobilized chemical gradient profiles, a result that opens important opportunities for understanding and controlling haptotactic behaviors, such as directed migration, that are key to various tissue regeneration applications.


Incidence, treatment, and outcomes of iron deficiency after laparoscopic Roux-en-Y gastric bypass: a 10-year analysis.
Laparoscopic Roux-en-Y gastric bypass (LRYGB) can lead to iron malabsorption through exclusion of the duodenum and proximal jejunum, decreased gastric acidity, and modified diet. Intravenous (IV) iron is a treatment for severe iron deficiency, but the incidence of iron deficiency and the frequency of treatment with IV iron after LRYGB are largely unknown. Our objective was to determine the incidence of iron deficiency and the frequency of IV iron administration after LRYGB. After obtaining IRB approval, the medical records of patients who underwent LRYGB from September 2001 to December 2011 were retrospectively reviewed. Inclusion criteria consisted of determination of at least 1 ferritin value after surgery. Patients were grouped by level of iron deficiency. Patients with at least 1 ferritin <50 ng/mL were considered iron deficient. Statistical analysis included ANOVA. There were 959 patients included; 84.9% were female. Mean age was 43.8 years, and preoperative body mass index was 47.4 kg/m(2). Four hundred ninety-two (51.3%) patients were iron deficient. Of these, 40.9% were severely iron deficient, with a ferritin <30 ng/mL. Intravenous iron was required by 6.7%. After IV iron therapy, 53% had improvement in hemoglobin and ferritin values, and 39% had improvement in ferritin values only. Given the incidence of iron deficiency after LRYGB observed in our series, patients should have iron status monitored carefully by all providers and be appropriately referred for treatment. Female patients should be counseled that there is a 50% chance they will become iron deficient after LRYGB.


The social and cultural life of the prisoners in the Jewish forced labor camp at Skarzysko-Kamienna.
The article discusses the social and cultural life among the Jewish prisoners at the Skarzysko-Kamienna forced labor camp in central Poland. The activities included organized observances of religious and national customs and creative arts and performances of many kinds. The activities served several functions for the artists and audiences. Whether as a source of income in the struggle for physical survival, or as a psychological escape from the terrible reality of daily life in the camp, or as a unifying element for the fragmented prisoner population, or as a from of political resistance, these social and cultural activities in the camp constitute a singular phenomenon in the history of the Holocaust.


[Clinical inertia in geriatrics].
Therapeutic inertia is a phenomenon with multiple etiologies, characterized by a "lack of initiation or modification of therapy when appropriate" and mainly affecting common chronic diseases (hypertension, diabetes, dyslipidemia). Caused by various processes including the lack of clinical evidence for the elderly, it can lead, in the elderly fragile by nature, to inappropriate prescribing. In a multidisciplinary approach, integrating the expertise of a pharmacist in the health care team improves the pharmacotherapeutic management of the geriatric patient.


Retuning mortality risk prediction in paediatric cardiac surgery: the additional role of early postoperative metabolic and respiratory profile.
The existing risk stratification scores for paediatric patients undergoing cardiac surgery include the Aristotle Basic Complexity (ABC) Score, the Risk Adjustment in Congenital Heart Surgery-1 (RACHS-1) Score and the Aristotle Comprehensive Complexity (ACC) Score. They are all based on the nature of the surgical operation (ABC and RACHS-1 Scores) with possible adjustment for a number of patient conditions (ACC Score). The present study investigates if the early postoperative parameters may be used to improve the preoperative mortality risk prediction. A retrospective study on 1392 consecutive patients aged ≤12 years old, undergoing cardiac surgery with cardiopulmonary bypass and without a residual right-to-left shunt was conducted. The ABC Score and metabolic and respiratory postoperative parameters at arrival in the intensive care unit were tested for association and discriminative power for operative mortality. The ABC yielded a c-statistic of 0.746. Additional independent predictors of operative mortality were postoperative hypoxia [Formula: see text] and arterial blood lactates. In a multivariable model including the ABC Score, postoperative hypoxia and arterial blood lactates remained independently associated with operative mortality. A modified ABC Score was created, consisting of the ABC Score plus 1.5 points in case of postoperative hypoxia plus 1 point per each 1 mmol/l of arterial blood lactates. The new model was significantly (P = 0.043) more discriminative than the ABC Score, with a c-statistic of 0.803. Early postoperative respiratory and metabolic parameters increased the accuracy and discrimination of the ABC Score. An external validation is needed to confirm our results.


Perspective: the potential of student organizations for developing leadership: one school's experience.
Leadership development is vital to the future of medicine. Some leadership development may take place through the formal curriculum of the medical school, yet extracurricular activities, such as student government and affiliated student organizations, can provide additional, highly valuable leadership development opportunities. These organizations and their missions can serve as catalysts for students to work with one another, with the faculty and administration of the medical school, with the community, and with local, regional, and national organizations. The authors have organized this discussion of the leadership development potential of student organizations around six important principles of leadership: ownership, experience, efficacy, sense of community, service learning, and peer-to-peer mentoring. They provide practical examples of these leadership principles from one institution. They do not presume that the school is unique, but they do believe their practical examples help to illuminate the potential of extracurricular programs for enhancing the leadership capabilities of future physicians. In addition, the authors use their examples to demonstrate how the medical school, its surrounding community, and the profession of medicine can benefit from promoting leadership through student organizations.


Targeted Thromboelastographic (TEG) Blood Component and Pharmacologic Hemostatic Therapy in Traumatic and Acquired Coagulopathy.
Trauma-induced coagulopathy (TIC) is a recently described condition which traditionally has been diagnosed by the common coagulation tests (CCTs) such as prothrombin time/international normalized ratio (PT/INR), activated partial thromboplastin time (aPTT), platelet count, and fibrinogen levels. The varying sensitivity and specificity of these CCTs have led trauma coagulation researchers and clinicians to use Viscoelastic Tests (VET) such as Thromboelastography (TEG) to provide Targeted Thromboelastographic Hemostatic and Adjunctive Therapy (TTHAT) in a goal directed fashion to those trauma patients in need of hemostatic resuscitation. This review describes the utility of VETs, in particular, TEG, to provide TTHAT in trauma and acquired non-trauma-induced coagulopathy.


Evidence for reduced and dysregulated turnover of dopamine in schizophrenia.
The dopamine (DA) hypothesis of schizophrenia proposes that schizophrenia is related to dopaminergic hyperactivity. To examine this hypothesis, this article reviews studies that have measured levels of DA and its metabolites in schizophrenic patients. From these studies, the following conclusions about schizophrenia emerge: (1) levels of DA and its metabolites can be highly variable; (2) DA turnover appears to be reduced in a subgroup of schizophrenic patients characterized as more chronic and treatment refractory, compared with normal controls; (3) DA turnover has been correlated positively with acute symptomatology and negatively with signs of chronic illness; and (4) central DA levels appear to be elevated. These conclusions support the author's hypothesis that DA turnover may be both reduced and dysregulated in schizophrenia. It is speculated that reduced turnover is primarily due to a deficiency in DA release and that dysregulation may result from the disruption of feedback mechanisms. Acute psychosis may be associated with a relative increase in the release of DA impinging on supersensitive postsynaptic receptors made so by chronic synaptic depletion of the transmitter.


Providing deterministic quality of service in slotted optical networks.
This paper proposes an efficient framework for deterministic service guarantees in slot-based optical networks. The framework uses the combination of a control plane and a data plane to solve the complex problem of capacity allocation, slot-matching and traffic scheduling. The control plane implements admission control and capacity allocation to source-destination node pairs and the data plane handles traffic aggregation, buffering, and scheduling. We propose an efficient algorithm in the control plane for slot collision-resolution. In the data plane, we present a comprehensive aggregation and scheduling mechanism that realizes Service Curves assurance. We use the Time-Domain Wavelength Interleaved Network (TWIN) architecture for the proof of concept and conduct extensive simulations to assess the performance of the algorithm and scheduling mechanism.


Brain-wide circuit-specific targeting of astrocytes.
Astrocytes are integral components of brain circuitry. They enwrap synapses, react to neuronal activity, and regulate synaptic transmission. Astrocytes are heterogeneous and exhibit distinct features and functions in different circuits. Selectively targeting the astrocytes associated with a given neuronal circuit would enable elucidation of their circuit-specific functions but has been technically challenging to date. Recently, we constructed anterograde transneuronal viral vectors based on yellow fever vaccine YFV-17D. Among them, the replication-incompetent YFVΔNS1-Cre can selectively turn on reporter genes in postsynaptic neurons if the viral gene NS1 is expressed in postsynaptic neurons. Here we show that without exogenous expression of NS1 at the postsynaptic sites, locally injected YFVΔNS1-Cre selectively turns on reporter genes in astrocytes in downstream brain regions. The targeting of astrocytes can occur across the whole brain but is specific for the neuronal circuits traced. Therefore, YFVΔNS1-Cre provides a tool for selective genetic targeting of astrocytes to reveal their circuit-specific roles.


Evaluation of an IgE ELISA with Culicoides spp. extracts and recombinant salivary antigens for diagnosis of insect bite hypersensitivity in Warmblood horses.
Insect bite hypersensitivity (IBH) in horses represents an immunoglobulin E (IgE)-mediated hypersensitivity to salivary antigens from biting midges (Culicoides spp.). The aim of this study was to evaluate and compare the performances of IgE ELISAs using recombinant Culicoides spp. Obsoletus group salivary gland antigens or crude whole body extracts ('ObsWBE'), C. nubeculosus recombinant proteins (Culn1, 3, 4, 5, 7, 8 and 10) and Obsoletus group recombinant proteins (Culo1 and 2). IgE levels were measured in plasma of 343 Warmblood horses classified as IBH-affected (n=167) and IBH-unaffected (n=176) according to the owners' descriptions. IBH-affected horses were subdivided based on the severity of their clinical signs at sampling and whether or not their IBH history was considered to be classical. The accuracies of the tests increased when clinical signs at sampling were more pronounced or when the IBH history could be considered as classical. A combination of IgE levels against the three best performing Culicoides spp. recombinant proteins (Culn4, Culo1 and Culo2) and ObsWBE resulted in the best performing test. When IBH-affected horses showing a classical history of the disease and severe clinical signs were compared with IBH-unaffected horses, the Youden's index at the optimal cut-off for the three tests in combination was 0.67. This optimal cut-off had a sensitivity of 70%, a specificity of 97% and a total accuracy of 92%. The performance of the IgE ELISA was affected by the severity of IBH clinical signs at sampling and was improved when IgE levels against several recombinant proteins were combined.


Stingray injuries.
Stingray injuries to humans are common in warm coastal areas. Wounds have a traumatic (puncture) component and a toxic (envenomation) component. The puncture component is like a stiletto-type knife wound, most often inflicted on the lower leg (waders) or arm (fishermen). Rare puncture injuries to the thorax or abdomen can cause serious injuries and death. Envenomation causes intense pain that is out of proportion to the apparent injury. The pain of stingray envenomation is relieved with hot-water immersion of the injured part, although the exact mechanism for this well-established phenomenon remains unclear. Current recommendations for treating stingray wounds are reviewed.


Genome-based vaccine development: a short cut for the future.
Bacterial infectious diseases remain a major cause of deaths and disabilities in the world. Although conventional vaccinology approaches were successful in conferring protection against several diseases, they failed in providing efficient vaccines against many others. Together to the sequencing of the first genome, a new chapter in the vaccinology history started to be written. Reverse vaccinology changed the way to think about vaccine development, using the information provided by the microorganisms' genome against themselves. Since then, reverse vaccinology has evolved and helped researchers to overcome the limits of the conventional vaccinology approaches and led to the discovery and development of novel vaccines concerning emerging diseases, like Neisseria meningitidis B and Streptococcus agalactiae. A lot of work must be done, but deciphering the information provided by genome sequences and using it to better understand the host-pathogen interactions has proved to be the key for protection.


Pathogenicity and oxidative stress in Nile tilapia caused by Aphanomyces laevis and Phoma herbarum isolated from farmed fish.
Identified (n = 17) and unidentified (n = 1) fish-pathogenic fungal species from 10 genera of Oomycetes and soil fungi were isolated from 40 infected freshwater fish samples of the species Oreochromis niloticus niloticus (Nile tilapia) and Clarias gariepinus (African catfish). Samples were collected from various fish farms in the Nile Delta, Egypt. Nile tilapia were tested in aquaria for their susceptibility to the commonest Oomycetes species, Aphanomyces laevis and Achlya klebsiana, and also against the 2 most prevalent pathogenic soil fungi, Paecilomyces lilacinus and Phoma herbarum. Two techniques were used: water bath exposure and intramuscular (subcutaneous) injection. Water bath exposure to the 2 species of Oomycetes caused greater mortalities of O. niloticus niloticus than intramuscular injection, but the reverse was true of the soil fungal species. Regardless of the infection method, the 2 Oomycetes species were more potent pathogens than the soil fungal species. In both gills and mytomal muscles of fish infected by A. laevis and P. herbarum, we measured and compared with controls the oxidative stress parameters total peroxide (TP), lipid peroxidation (LPO) and nitric oxide (NO), as well as levels of the antioxidants vitamin E and glutathione (GSH), and superoxide dismutase (SOD) and catalase (CAT) activities. Infection by these 2 fungal species through either spore suspension or spore injection significantly increased oxidative damage in gills and induced marked decrease in most studied antioxidants. In addition, both routes showed similar effects and A. laevis depressed the antioxidants CAT, vitamin E and GSH more than P. herbarum.


[Innovating the service management of public health: the experience of the Barcelona Public Health Agency (Spain)].
We describe the evolution of the organization of public health services in the city of Barcelona (Catalonia, Spain) until the creation of the Barcelona Public Health Agency. This Agency is a consortium created by the Barcelona City Council and the Government of Catalonia as the sole entity responsible for regional and local public health services in the city. The underlying logic for the Agency's design, as well as its mission, vision and value statements, strategy, services' portfolio, and the role of leadership in the process, are analyzed. Aspects related to the Agency's quality and communication plans, as well as the design of its processes, and its policy in terms of alliances for research and training in public health, are discussed. Finally, the main challenges for the future are described.


Social Media Use, Unhealthy Lifestyles, and the Risk of Miscarriage Among Pregnant Women During the COVID-19 Pandemic: Prospective Observational Study.
The COVID-19 pandemic has resulted in changes to normal life and disrupted social and economic function worldwide. However, little is known about the impact of social media use, unhealthy lifestyles, and the risk of miscarriage among pregnant women during the COVID-19 pandemic. This study aims to assess the association between social media use, unhealthy lifestyles, and the risk of miscarriage among pregnant women in the early stage of the COVID-19 pandemic in China. In this prospective cohort study, 456 singleton pregnant women in mainland China were recruited during January and February 2020. Sociodemographic characteristics, history of previous health, social media use, and current lifestyles were collected at baseline, and we followed up about the occurrence of miscarriage. Log-binomial regression models were used to estimate the risk ratios (RRs) of miscarriage for women with different exposures to COVID-19-specific information. Among all the 456 pregnant women, there were 82 (18.0%) who did no physical activities, 82 (18.0%) with inadequate dietary diversity, 174 (38.2%) with poor sleep quality, and 54 (11.8%) spending >3 hours on reading COVID-19 news per day. Women with excessive media use (>3 hours) were more likely to be previously pregnant (P=.03), have no physical activity (P=.003), have inadequate dietary diversity (P=.03), and have poor sleep quality (P<.001). The prevalence of miscarriage was 16.0% (n=73; 95% CI 12.6%-19.4%). Compared with women who spent 0.5-2 hours (25/247, 10.1%) on reading COVID-19 news per day, miscarriage prevalence in women who spent <0.5 hours (5/23, 21.7%), 2-3 hours (26/132, 19.7%), and >3 hours (17/54, 31.5%) was higher (P<.001). Miscarriage prevalence was also higher in pregnant women with poor sleep quality (39/174, 22.4% vs 34/282, 12.1%; P=.003) and a high education level (66/368, 17.9% vs 7/88, 8.0%; P=.02). In the multivariable model, poor sleep quality (adjusted RR 2.06, 95% CI 1.24-3.44; P=.006), 2-3 hours of media use daily (adjusted RR 1.74, 95% CI 1.02-2.97; P=.04), and >3 hours of media use daily (adjusted RR 2.56, 95% CI 1.43-4.59; P=.002) were associated with miscarriage. In the sensitivity analysis, results were still stable. Pregnant women with excessive media use were more likely to have no physical activity, inadequate dietary diversity, and poor sleep quality. Excessive media use and poor sleep quality were associated with a higher risk of miscarriage. Our findings highlight the importance of healthy lifestyles during the COVID-19 pandemic.


Combination of scoring functions improves discrimination in protein-protein docking.
Two structure-based potentials are used for both filtering (i.e., selecting a subset of conformations generated by rigid-body docking), and rescoring and ranking the selected conformations. ACP (atomic contact potential) is an atom-level extension of the Miyazawa-Jernigan potential parameterized on protein structures, whereas RPScore (residue pair potential score) is a residue-level potential, based on interactions in protein-protein complexes. These potentials are combined with other energy terms and applied to 13 sets of protein decoys, as well as to the results of docking 10 pairs of unbound proteins. For both potentials, the ability to discriminate between near-native and non-native docked structures is substantially improved by refining the structures and by adding a van der Waals energy term. It is observed that ACP and RPScore complement each other in a number of ways (e.g., although RPScore yields more hits than ACP, mainly as a result of its better performance for charged complexes, ACP usually ranks the near-native complexes better). As a general solution to the protein-docking problem, we have found that the best discrimination strategies combine either an RPScore filter with an ACP-based scoring function, or an ACP-based filter with an RPScore-based scoring function. Thus, ACP and RPScore capture complementary structural information, and combining them in a multistage postprocessing protocol provides substantially better discrimination than the use of the same potential for both filtering and ranking the docked conformations.


Fatal and nonfatal injuries involving fishing vessel winches--Southern shrimp fleet, United States, 2000-2011.
Workers in the commercial fishing industry have the highest occupational fatality rate in the United States, nearly 35 times higher in 2011 than the rate for all U.S. workers. During 2000-2009, a total of 504 fishermen were killed in the U.S. fishing industry, most commonly by drowning as a result of vessels sinking (51%) and falls overboard (30%). Another 10% of fatalities (51 deaths) were caused by injuries sustained onboard vessels, such as entanglement in machinery. This type of fatality occurred most often in the Gulf of Mexico. To analyze fatal and nonfatal injuries involving deck winches in the Southern shrimp fleet during 2000-2011, CDC obtained data from its Commercial Fishing Incident Database and the U.S. Coast Guard. Injury patterns were examined, and risk ratios (RRs) were calculated to compare the probability of fatal outcomes from incidents involving different winch mechanisms and operating situations. During 2000-2011, eight fatal and 27 work-related injuries involving deck winches occurred in the Southern shrimp fleet, which operates in the Gulf of Mexico and off the Atlantic coast from Florida to North Carolina. Injuries involving the winch drum had a higher risk for fatal outcomes compared with injuries involving the winch cathead. Fatal outcomes also were associated with being alone on the vessel and being alone on deck. Interventions to prevent deck winch injuries might include guarding of winch drums and catheads, avoiding working alone on deck, not wearing baggy clothing, and improvements to cable winding guides. Training of deckhands in first aid and emergency procedures might reduce the severity of injuries when entanglements occur.


Intravenous fish oil lipid emulsion promotes a shift toward anti-inflammatory proresolving lipid mediators.
Parenteral nutrition (PN)-associated liver disease (PNALD) is a life-threatening complication of the administration of PN. The development of PNALD may be partly due to the composition of the lipid emulsion administered with PN: soybean oil-based lipid emulsions (SOLE) are associated with liver disease, while fish oil-based lipid emulsions (FOLE) are associated with prevention and improvement of liver disease. The objective of this study was to determine how the choice of lipid emulsion modified the production of bioactive lipid mediators (LMs). We utilized a mouse model of steatosis to study the differential effect of FOLE and SOLE. We subsequently validated these results in serum samples from a small cohort of human infants transitioning from SOLE to FOLE. In mice, FOLE was associated with production of anti-inflammatory, proresolving LMs; SOLE was associated with increased production of inflammatory LMs. In human infants, the transition from SOLE to FOLE was associated with a shift toward a proresolving lipidome. Together, these results demonstrate that the composition of the lipid emulsion directly modifies inflammatory homeostasis.


Novel sample-substrates for the determination of new psychoactive substances in oral fluid by desorption electrospray ionization-high resolution mass spectrometry.
A reliable screening and non invasive method based on the use of microextraction by packed sorbent coupled with desorption electrospray ionization-high resolution mass spectrometry was developed and validated for the detection of new psychoactive substances in oral fluid. The role of different sample substrates in enhancing signal intensity and stability was evaluated by testing the performances of two polylactide-based materials, i.e. non-functionalized and functionalized with carbon nanoparticles, and a silica-based material compared to commercially available polytetrafluorethylene supports. The best results were achieved by using the non-functionalized polylactide substrates to efficiently ionize compounds in positive ionization mode, whereas the silica coating proved to be the best choice for operating in negative ionization mode. LLOQs in the low μg/L, a good precision with CV% always lower than 16% and RR% in the 83(±4)-120(±2)% range, proved the suitability of the developed method for the determination of the analytes in oral fluid. Finally, the method was applied for screening oral fluid samples for the presence of psychoactive substances during private parties, revealing mephedrone in only one sample out of 40 submitted to analysis.


High prevalence of hypovitaminosis D in pregnant Japanese women with threatened premature delivery.
Serum 25-hydroxyvitamin D (25-OHD) concentrations are thought to accurately reflect vitamin D stores, and vitamin D deficiency causes secondary hyperparathyroidism, irreversible bone loss, and increased risk of fracture. Recent studies suggest that decrease of serum 25-OHD level in mothers could increase the risk of preeclampsia, cesarean section, and craniotabes. Furthermore, this deficiency may affect bone mass and the incidence of neuromuscular diseases of their children in the future. In the present study, the serum concentration of 25-OHD in 93 pregnant women after the 30th week of their gestation was determined by direct radioimmunoassay. Mean 25-OHD levels in spring, summer, fall, and winter were 14.3 ± 5.1, 15.7 ± 6.4, 13.7 ± 5.1, and 13.9 ± 4.2 ng/ml, respectively. Severe vitamin D deficiency (25-OHD < 10 ng/ml) was found in 10 of these 93 women. Overall, hypovitaminosis D, which was defined as serum 25-OHD concentration equal to or less than 20 ng/ml, was revealed in 85 mothers (89.5%). Serum 25-OHD levels were not associated with either intact parathyroid hormone or corrected calcium concentrations, but were negatively associated with serum type I collagen N-terminal telopeptide and bone-specific alkaline phosphatase in these subjects. Mothers with threatened premature delivery had significantly lower 25-OHD levels (11.2 ± 3.2 ng/ml) than those in mothers with normal delivery (15.6 ± 5.1 ng/ml). In conclusion, the present data suggest a high prevalence of hypovitaminosis D in perinatal pregnant Japanese women throughout the year, which seems to affect bone metabolism and to be associated with threatened premature delivery.


On the use of a quality index to specify high energy photon beams.
It is important to specify the beam quality in a simple and nonambiguous way in order on one hand to make comparisons easier between treatments performed in various hospitals, or at different times in the same hospital and on the other hand to facilitate the choice of numerical values for factors like restricted mass-collision stopping-power ratios and perturbation correction factors used in the conversion of ionization measurements into absorbed dose. We have adopted for high-energy photon beam specification a quality index (I) defined by the ratio (I20/I10) of ionizations measured with a constant source-detector distance for a reference field size 10 X 10 cm2. We have found that this quality index is independent of the source detector distance. On the other hand, the apparent linear attenuation coefficient measured on the exponential part of the tissue-maximum ratio curve can be calculated for any field size from the value of I for most high energy photon beams. In order to check the validity of the quality index for other linacs from other manufacturers, we have compared our results to published data related to various photon beams in a wide energy range: 2.5 to 45 MV.


Robust initial detection of landmarks in film-screen mammograms using multiple FFDM atlases.
Automated analysis of mammograms requires robust methods for pectoralis segmentation and nipple detection. Locating the nipple is especially important in multiview computer aided detection systems, in which findings are matched across images using the nipple-to-finding distance. Segmenting the pectoralis is a key preprocessing step to avoid false positives when detecting masses due to the similarity of the texture of mammographic parenchyma and the pectoral muscle. A multiatlas algorithm capable of providing very robust initial estimates of the nipple position and pectoral region in digitized mammograms is presented here. Ten full-field digital mammograms, which are easily annotated attributed to their excellent contrast, are robustly registered to the target digitized film-screen mammogram. The annotations are then propagated and fused into a final nipple position and pectoralis segmentation. Compared to other nipple detection methods in the literature, the system proposed here has the advantages that it is more robust and can provide a reliable estimate when the nipple is located outside the image. Our results show that the change in the correlation between nipple-to-finding distances in craniocaudal and mediolateral oblique views is not significant when the detected nipple positions replace the manual annotations. Moreover, the pectoralis segmentation is acceptable and can be used as initialization for a more complex algorithm to optimize the outline locally. A novel aspect of the method is that it is also capable of detecting and segmenting the pectoralis in craniocaudal views.


Use of Oxidized Regenerated Cellulose (ORC)/Collagen/Silver-ORC Dressings Alone or Subsequent to Advanced Wound Therapies in Complex Wounds.
Chronic or senescent wounds are difficult to heal and often require a multimodal treatment plan. Negative pressure wound therapy (NPWT) or advanced wound dressings, such as oxidized regenerated cellulose (ORC)/collagen/silver-ORC dressings, can be used to promote granulation tissue development and stimulate wound healing in these complex wounds. This article examines the use of ORC/collagen/silver-ORC dressings alone or subsequent to advanced wound therapies in a retrospective cohort of 24 patients. Wounds were assessed upon presentation. If necessary, oral and/or intravenous antibiotics were administered. Each wound underwent sharp debridement. Patients received either ORC/collagen/silver-ORC dressings with a secondary dressing alone or following NPWT. Skin substitutes and epidermal grafting also were utilized to promote wound healing and wound size reduction. Twenty-four patients with an average age of 66.8 ± 12.7 years were treated. The most prevalent comorbidities were hypertension, diabetes, obesity, peripheral neuropathy, hyperlipidemia, coronary heart disease, and tobacco use. Wound types (N = 27) included diabetic foot ulcers, surgical wounds, traumatic wounds, an ulcer (secondary to chronic gout with tophi), and thermal burns. All 27 wounds fully closed, with an average time to heal of 65.5 ± 41.5 days. Use of advanced treatment modalities including NPWT, epidermal grafting, and ORC/collagen/silver-ORC dressings contributed to wound healing in these patients with complex and/or chronic wounds.


Photoinduced intramolecular electron-transfer processes in [60]fullerene and N,N-Bis(biphenyl)aniline molecular systems in solutions.
Photoinduced intramolecular charge-separation and charge-recombination processes in covalently connected C(60)-(spacer)-bis(biphenyl)aniline (C(60)-sp-BBA) and C(60)-((spacer)-bis(biphenyl)aniline)(2) (C(60)-(sp-BBA)(2)) have been studied by time-resolved fluorescence and transient absorption methods. Since a flexible alkylthioacetoamide chain was employed as the spacer, the folded structures in which the BBA moiety approaches the C(60) moiety were obtained as optimized structures by molecular orbital calculations. The observed low fluorescence intensity and the short fluorescence lifetime of the C(60) moiety of these molecular systems indicated that charge separation takes place via the excited singlet state of the C(60) moiety in a quite fast rate and high efficiency even in the nonpolar solvent toluene, which was a quite new observation compared with reported dyads with different spacers. From the absorption bands at 880 and 1000 nm in the nanosecond transient absorption spectra, generations of C(60)(.-)-sp-BBA(.+) and C(60)(.-)-(sp-BBA(.+))(sp-BBA) were confirmed. The rates of charge separation and charge recombination for C(60)-(sp-BBA)(2) are faster than those for C(60)-sp-BBA, suggesting that one of the BBA moieties approaches the C(60) moiety by pushing another BBA moiety because of the flexible spacers.


Improving Endoscopic Adherence to Quality Metrics in Colonoscopy.
Appropriate documentation of quality metrics in the endoscopy reports provides evidence that a thorough and complete examination was performed. The aim of our study was to assess compliance with 3 current quality metrics for colonoscopy defined by the American Society for Gastrointestinal Endoscopy. We retrospectively examined colonoscopy reports from 6 gastroenterologists at Ochsner Medical Center for appropriate documentation of the quality of the bowel preparation and photodocumentation of the appendiceal orifice and the ileocecal valve. A performance review and educational session then took place with each physician. Subsequent colonoscopy reports were evaluated to monitor for improvement. Bowel preparation documentation was high before and after the educational sessions (97.5% and 97.2%). Preeducation, the mean photodocumentation rate of the appendiceal orifice was 55% (range, 23%-84%). For the ileocecal valve, the documentation rate was 32.5% (range, 3%-73%). Posteducation, the mean appendiceal orifice labeling increased to an average of 91%, with a median change of 28.5% (P=0.0313). Documentation of the ileocecal valve improved to an average of 73%, a median change of 37.5% (P=0.0625). Although reassessment of subsequent reports will be necessary to evaluate the permanence of this intervention, our evidence suggests that educational sessions can improve the quality and accuracy of documentation of quality metrics during colonoscopies.


A combined experimental and theoretical investigation on the role of halide ligands on the catecholase-like activity of mononuclear nickel(II) complexes with a phenol-based tridentate ligand.
Three new mononuclear nickel(II) complexes, namely, [NiL(1)(H2O)3]I2·H2O (1), [NiL(1)(H2O)3]Br2·H2O (2), and [NiL(1)(H2O)3]Cl2·2H2O (3) [HL(1) = 2-[(2-piperazin-1-ylethylimino)methyl]phenol], have been synthesized and structurally characterized. Structural characterization reveals that they possess similar structure: [NiL(1)(H2O)3](2+) complex cations, two halide counteranions, and lattice water molecules. One of the nitrogen atoms of the piperazine moiety is protonated to provide electrical neutrality to the system, a consequence observed in earlier studies (Inorg. Chem. 2010, 49, 3121; Polyhedron 2013, 52, 669). Catecholase-like activity has been investigated in methanol by a UV-vis spectrophotometric study using 3,5-di-tert-butylcatechol (3,5-DTBC) as the model substrate. Complexes 1 and 2 are highly active, but surprisingly 3 is totally inactive. The coordination chemistries of 1 and 2 remain unchanged in solution, whereas 3 behaves as a 1:1 electrolyte, as is evident from the conductivity study. Because of coordination of the chloride ligand to the metal in solution, it is proposed that 3,5-DTBC is not able to effectively approach an electrically neutral metal, and consequently complex 3 in solution does not show catecholase-like activity. Density functional theory (DFT) calculations corroborate well with the experimental observations and thus, in turn, support the proposed hypothesis of inactivity of 3. The cyclic voltametric study as well as DFT calculations suggests the possibility of a ligand-centered reduction at -1.1 V vs Ag/AgCl electrode. An electron paramagnetic resonance (EPR) experiment unambiguously hints at the generation of a radical from EPR-inactive 1 and 2 in the presence of 3,5-DTBC. Generation of H2O2 during catalysis has also been confirmed. DFT calculations support the ligand-centered radical generation, and thus a radical mechanism has been proposed for the catecholase-like activity exhibited by 1 and 2. Upon heating, 2 and 3 lose water molecules in two steps (first lattice waters, followed by coordinating water molecules), whereas 3 loses four water molecules in a single step, as revealed from thermogravimetric analysis. The totally dehydrated species are red, in all cases having square-planar geometry, and have amorphous nature, as is evident from a variable-temperature powder X-ray diffraction study.


Foxj3 regulates thermogenesis of brown and beige fat via induction of PGC-1α.
Enhancing the development and thermogenesis in brown and beige fat represents a potential treatment for obesity. In the present study, we show that Foxj3 expression in fat is stimulated by cold exposure and a β-adrenergic agonist. Adipose-specific Foxj3 knockout impairs the thermogenic function of brown fat, leading to morphological whitening of brown fat and obesity. Adipose Foxj3-deficient mice display increased fasting blood glucose levels and hepatic steatosis on a chow diet. Foxj3 deficiency inhibits the browning of inguinal white adipose tissue (iWAT) following β3-agonist treatment of mice. Furthermore, depletion of Foxj3 in primary brown adipocytes reduces the expression of thermogenic genes and cellular respiration, indicating that the Foxj3 effects on the thermogenic program are cell autonomous. In contrast, Foxj3 overexpression in primary brown adipocytes enhances the thermogenic program. Moreover, AAV-mediated Foxj3 overexpression in brown fat and inguinal WAT increases energy expenditure and improves systemic metabolism on either a chow diet or a high-fat diet. Finally, Foxj3 deletion in fat inhibits the β3 agonist-mediated induction of WAT browning and brown adipose tissue thermogenesis. Mechanistically, cold-inducible Foxj3 stimulates the expression of PGC-1α and UCP1, subsequently promoting energy expenditure. The present study identifies Foxj3 as a critical regulator of fat thermogenesis, and targeting Foxj3 in fat might be a therapeutic strategy for treating obesity and metabolic diseases.


Ursolic Acid Impairs Cellular Lipid Homeostasis and Lysosomal Membrane Integrity in Breast Carcinoma Cells.
Cancer is one of the leading causes of death worldwide, thus the search for new cancer therapies is of utmost importance. Ursolic acid is a naturally occurring pentacyclic triterpene with a wide range of pharmacological activities including anti-inflammatory and anti-neoplastic effects. The latter has been assigned to its ability to promote apoptosis and inhibit cancer cell proliferation by poorly defined mechanisms. In this report, we identify lysosomes as the essential targets of the anti-cancer activity of ursolic acid. The treatment of MCF7 breast cancer cells with ursolic acid elevates lysosomal pH, alters the cellular lipid profile, and causes lysosomal membrane permeabilization and leakage of lysosomal enzymes into the cytosol. Lysosomal membrane permeabilization precedes the essential hallmarks of apoptosis placing it as an initial event in the cascade of effects induced by ursolic acid. The disruption of the lysosomal function impairs the autophagic pathway and likely partakes in the mechanism by which ursolic acid kills cancer cells. Furthermore, we find that combining treatment with ursolic acid and cationic amphiphilic drugs can significantly enhance the degree of lysosomal membrane permeabilization and cell death in breast cancer cells.


Hypertrophic cardiomyopathy: low frequency of mutations in the beta-myosin heavy chain (MYH7) and cardiac troponin T (TNNT2) genes among Spanish patients.
Mutations in the cardiac beta-myosin heavy chain (MYH7) and cardiac troponin T (TNNT2) genes are reportedly responsible for up to 40% of familial cases with hypertrophic cardiomyopathy (HC). Although there are no mutational hotspots, most of the mutations are located in specific exons of the MYH7 and TNNT2 genes. Currently it is not possible to predict the phenotype in carriers of mutations in these genes, although it is widely accepted that mutations in the MYH7 gene predispose to severe HC, whereas TNNT2 mutations are frequently linked to sudden cardiac death (SCD) in spite of minimal hypertrophy. We sequenced exons 8, 9, 13-16, 19, 20, 22-24, and 30 of the MYH7 gene and exons 8, 9, 11, and 14-16 of the TNNT2 gene in 30 HC patients (18-60 years of age) from the region of Asturias (Northern Spain); 25 cases (80%) had a family history of the disease. Genomic DNA was amplified, and fragments were directly sequenced. Each DNA variant found in the patients was also analyzed in 200 healthy controls through single-strand conformation analysis. Four of the probands had nucleotide changes absent in the healthy controls. Two cases had mutations previously described in the MYH7 gene (exon 14, Arg453Cys) or the TNNT2 gene (exon 16, Arg278Cys). Two cases had new mutations (MYH7 exon 22, Met822Val; TNNT2 exon 14, Lys247Arg) not found among the healthy controls. We found MYH7 Met822Val in a woman with a severe form of HC; the mutation was absent in her parents, indicating a de novo mutation. MYH7 R453C was present in a woman with mild HC, mother of a son who died from SCD. TNNT2 R278C was present in a woman with severe HC, but a sister and a daughter were mutation carriers and did not have hypertrophy. A patient with severe HC was carrier of TNNT2 247Arg. Mutations in the MYH7 and TNNT2 genes can be found in patients without a family history of HC. However, compared with other populations MYH7 or TNNT2 mutations were rare among our HC patients. This study illustrates the extreme phenotypic heterogeneity in carriers of MYH7 or TNNT2 mutations.


Detection of dietary DNA, protein, and glyphosate in meat, milk, and eggs.
Products such as meat, milk, and eggs from animals that have consumed genetically engineered (GE) feed are not currently subject to mandatory GE labeling requirements. Some voluntary "non-genetically modified organism" labeling has been associated with such products, indicating that the animals were not fed GE crops, as there are no commercialized GE food animals. This review summarizes the available scientific literature on the detection of dietary DNA and protein in animal products and briefly discusses the implications of mandatory GE labeling for products from animals that have consumed GE feed. Because glyphosate is used on some GE crops, the available studies on glyphosate residues in animal products are also reviewed. In GE crops, recombinant DNA (rDNA) makes up a small percentage of the plant's total DNA. The final amount of DNA in food/feed depends on many factors including the variable number and density of cells in the edible parts, the DNA-containing matrix, environmental conditions, and the specific transgenic event. Processing treatments and animals' digestive systems degrade DNA into small fragments. Available reports conclude that endogenous DNA and rDNA are processed in exactly the same way in the gastrointestinal tract and that they account for a very small proportion of food intake by weight. Small pieces of high copy number endogenous plant genes have occasionally been detected in meat and milk. Similarly sized pieces of rDNA have also been identified in meat, primarily fish, although detection is inconsistent. Dietary rDNA fragments have not been detected in chicken or quail eggs or in fresh milk from cows or goats. Collectively, studies have failed to identify full-length endogenous or rDNA transcripts or recombinant proteins in meat, milk, or eggs. Similarly, because mammals do not bioaccumulate glyphosate and it is rapidly excreted, negligible levels of glyphosate in cattle, pig and poultry meat, milk, and eggs have been reported. Despite consumer concern about the presence of trace concentrations of glyphosate that might have been applied to feed crops and/or the presence of rDNA or recombinant proteins in meat, milk, and eggs, the available data do not provide evidence to suggest that products from animals that have consumed approved GE feed crops differ in any distinguishable way from those derived from animals fed conventional feed or that products from animals fed GE feedstuffs pose novel health risks.


Normality sensing licenses local T cells for innate-like tissue surveillance.
The increasing implication of lymphocytes in general physiology and immune surveillance outside of infection poses the question of how their antigen receptors might be involved. Here, we show that macromolecular aggregates of intraepidermal γδ T cell antigen receptors (TCRs) in the mouse skin aligned with and depended on Skint1, a butyrophilin-like (BTNL) protein expressed by differentiated keratinocytes (KCs) at steady state. Interruption of TCR-mediated 'normality sensing' had no impact on γδ T cell numbers but altered their signature phenotype, while the epidermal barrier function was compromised. In addition to the regulation of steady-state physiology, normality sensing licensed intraepidermal T cells to respond rapidly to subsequent tissue perturbation by using innate tumor necrosis factor (TNF) superfamily receptors. Thus, interfering with Skint1-dependent interactions between local γδ T cells and KCs at steady state increased the susceptibility to ultraviolet B radiation (UVR)-induced DNA damage and inflammation, two cancer-disposing factors.