N-Glycans on EGF domain-specific O-GlcNAc transferase (EOGT) facilitate EOGT maturation and peripheral endoplasmic reticulum localization.
Epidermal growth factor (EGF) domain-specific O-GlcNAc transferase (EOGT) is an endoplasmic reticulum (ER)-resident protein that modifies EGF repeats of Notch receptors and thereby regulates Delta-like ligand-mediated Notch signaling. Several EOGT mutations that may affect putative N-glycosylation consensus sites are recorded in the cancer database, but the presence and function of N-glycans in EOGT have not yet been characterized. Here, we identified N-glycosylation sites in mouse EOGT and elucidated their molecular functions. Three predicted N-glycosylation consensus sequences on EOGT are highly conserved among mammalian species. Within these sites, we found that Asn-263 and Asn-354, but not Asn-493, are modified with N-glycans. Lectin blotting, endoglycosidase H digestion, and MS analysis revealed that both residues are modified with oligomannose N-glycans. Loss of an individual N-glycan on EOGT did not affect its endoplasmic reticulum (ER) localization, enzyme activity, and ability to O-GlcNAcylate Notch1 in HEK293T cells. However, simultaneous substitution of both N-glycosylation sites affected both EOGT maturation and expression levels without an apparent change in enzymatic activity, suggesting that N-glycosylation at a single site is sufficient for EOGT maturation and expression. Accordingly, a decrease in O-GlcNAc stoichiometry was observed in Notch1 co-expressed with an N263Q/N354Q variant compared with WT EOGT. Moreover, the N263Q/N354Q variant exhibited altered subcellular distribution within the ER in HEK293T cells, indicating that N-glycosylation of EOGT is required for its ER localization at the cell periphery. These results suggest critical roles of N-glycans in sustaining O-GlcNAc transferase function both by maintaining EOGT levels and by ensuring its proper subcellular localization in the ER.


Monocyte-to-High Density Lipoprotein Cholesterol Ratio Positively Predicts Coronary Artery Disease and Multi-Vessel Lesions in Acute Coronary Syndrome.
We investigated the hypothesis that MHR (monocyte-to-high density lipoprotein cholesterol ratio) is related to the severity of coronary artery in ACS (acute coronary syndrome). In this case-control study, we recruited 15,853 participants undergoing the first time percutaneous coronary intervention (PCI) including 4093 normal controls, 10,518 chronic coronary artery disease (CAD), and 1242 ACS cases. Examination of demographic clinical data and biochemical profiles, as well as MHR values, were performed before PCI. The relationship between MHR and severity of coronary artery lesion in ACS was analyzed. We also used a flow cytometric assay to distinguish CD14+/CD16- classical monocyte subsets in peripheral blood mononucleated cells from CAD patients. MHR was higher in patients with ACS compared with MHR in normal control and chronic CAD (normal control vs chronic CAD vs ACS: 0.46 ± 0.27 × 109/mmol vs 0.53 ± 0.29 × 109/mmol vs 0.73 ± 0.47 × 109/mmol, P < 0.001). MHR showed a significantly progressive increase as the angiographic severity of coronary lesions increased (single vessel lesion vs multi-vessel lesions in ACS: 0.54 ± 0.31 × 109/mmol vs 0.58 ± 0.35 × 109/mmol, P < 0.001), and classical monocyte subset to HDL-C ratio (CMHR) was increased in with CAD patients compared with control [4.69 (IQR, 1.06, 2.97) × 103/mmol vs 1.92 (IQR, 0.92, 3.04) × 103/mmol, P = 0.02]. Using a multivariate analysis, after adjusting for age, gender, body mass index (BMI), diabetes, and dyslipidemia, MHR was positively associated with multi-vessel lesions in ACS [OR (odds ratio): 1.28 (95% CI: 1.03-1.59, P = 0.029)]. MHR level could be a potential predictor of coronary artery lesion severity in ACS.


Comparative oral dose toxicokinetics of sodium selenite and selenomethionine.
Selenium (Se) poisoning by different forms of Se occurs in the United States. However, the toxicokinetics of different selenocompounds after oral ingestion is not well documented. In this study the toxicokinetics of Se absorption, distribution and elimination were determined in serum and whole blood of lambs that were orally dosed with increasing doses of Se as sodium selenite (inorganic Se) or selenomethionine (SeMet, organic Se). Thirty-two lambs were randomly assigned to eight treatment groups, with four animals per group. Se was administered at 1, 2 or 3 mg kg-1 body weight, as either sodium selenite or SeMet with proper control groups. Blood and serum were collected at predetermined time points for 7 days post-dosing. Resulting Se concentrations in both serum and whole blood from SeMet treatment groups were significantly greater than those given equimolar doses of Se as sodium selenite. Se concentrations in serum and whole blood of lambs dosed with SeMet peaked at significantly greater concentrations when compared with lambs dosed with equimolar doses of sodium selenite. Based on the serum and whole blood kinetics, the rate of Se absorption was greater for SeMet than for sodium selenite although rates of absorption for both Se forms decreased with increasing dose. The rates of Se elimination increased with dose. These results demonstrate that SeMet has a greater absorption rate and a similar retention time resulting in a greater area under the curve and thus bioavailability than sodium selenite, which must be considered in both overdose and nutritional exposures. Published 2016. This article is a U.S. Government work and is in the public domain in the USA.


Ganglioside-modulated protein phosphorylation. Partial purification and characterization of a ganglioside-stimulated protein kinase in brain.
Gangliosides have profound modulatory effects on protein phosphorylation in brain. A protein kinase activated directly by gangliosides has been partially purified from the particulate fractions of guinea pig brain through extraction with nonionic detergent, ion-exchange chromatography, hydrophobic chromatography, and gel filtration. This novel ganglioside-stimulated protein kinase is distinct from cAMP-dependent, Ca2+/calmodulin-dependent, and Ca2+/phospholipid-dependent protein kinases. The partially purified kinase preparation could undergo ganglioside-stimulated autophosphorylation of a major phosphoprotein with Mr corresponding to 68,000. It also could phosphorylate exogenous substrates such as the synthetic peptide Leu-Arg-Arg-Ala Ser-Leu-Gly. The requirement of gangliosides for the activation of kinase activity is dose-dependent and specific. Among the various gangliosides tested, GT1b and GD1a were found to be the most potent activators, whereas GD1b and GM1 were slightly less effective. The activation process is rapid and does not require the presence of Ca2+, suggesting that the stimulatory effect of gangliosides is not mediated through limited proteolysis or Ca2+-glycolipid complexes. Although the exact physiological significance of the ganglioside-stimulated protein kinase is not known at present, it is possible that certain functions related to gangliosides in the nervous system are mediated through the activation of this novel enzyme.


Chondroblastoma Affecting the Apophysis of the Greater Trochanter in a Child.
Chondroblastomas are rare primary bone tumours typically affecting the epiphyses and less frequently the apophyses of the growing skeleton. Most cases are treated by intralesional curettage with or without local adjuvants and this technique can produce good long-term outcomes. Herein, we describe a case of chondroblastoma of the greater trochanter in a 12-year-old male child that was treated by intralesional curettage and grafting with calcium phosphate bone cement (Neocement Inject® P, Bioceramed, Loures, Portugal). A brief review of the literature is also presented.


Compact, UAV-mounted hyperspectral imaging system with automatic geometric distortion rectification.
A highly compact hyperspectral imager with an automatic geometric rectification function is developed in this study, which can be mounted on a UAV for ultra-wide range hyperspectral imaging. For better application, the system can provide visible light image transmission and hyperspectral imaging in the real-time mode. A specific design is proposed to allow the visible light camera and hyperspectral camera to share the same telescope optical path, making the system have a high integration level with a total mass of 1.9 kilograms. Thanks to the sharing-optical-path design, the field of view (FOV), frame rate, and spatial resolution are modified the same between the visible light camera and hyperspectral camera. As a result, the geometric rectification is easily performed, and repeated rectifications are eliminated to improve the imaging efficiency. A FOV of 40 degrees in the frame direction and 26 degrees in the flight direction are realized with a focal length of 13mm, providing a large spectral range from 400 nm to 1000 nm and an excellent spectral resolution of 2.5 nm. An automatic geometric rectification workflow is presented and verified in experiments, which can improve the geometric rectification of hyperspectral images in the presence of low-quality UAV navigation data through the incorporation of frame images. Experimental results show that the relative accuracy of geometric rectification is less than 2 pixels when applying the algorithm to our system dataset.


Single-molecule conductance investigation of BDT derivatives: an additional pattern found to induce through-space channels beyond π-π stacking.
The through-space conductance of individual molecules is supposed to improve the macroscopic carrier movement, but the most widely acclaimed through-space conductance channel just existed in sufficiently close π-π stacked benzene rings. As a breakthrough to this primary cognition, additional conducting channels were confirmed to exist in non-strict face-to-face aligned thiophenes or phenyl-thiophene in BDT derivatives for the first time.


Sclerochoroidal calcification associated with Albright's hereditary osteodystrophy.
A 47-year-old woman presented with bilateral gradual loss of vision, ocular discomfort and seeing a black shadow in her right visual field over 6 months duration. Her medical history was extensive including: developmental delay, pseudohypoparathyroidism, hypertension, spinal stenosis, epilepsy and suspected idiopathic intracranial hypertension. Ocular examination revealed choroidal elevation in both eyes, which were highly ecogenic on ecography and confirmed to be calcifications of choroids on CT scan in her both eyes. She had subnormal vision and reduced colour vision in her both eyes. Electrodiagostic studies suggested rod dysfunction. She had typical features of Albright's hereditary dystrophy and was positive for the GNAS 1 mutation. She is currently being monitored by ophthalmologlists and is also under a medical team undergoing further assessment with regard to her treatment.


A genetic polymorphism in lincRNA-uc003opf.1 is associated with susceptibility to esophageal squamous cell carcinoma in Chinese populations.
The development of esophageal squamous cell carcinoma (ESCC) is a multifactorial process, and associations between genetic variants and ESCC have been identified in genome-wide association studies. The aim of this study was to evaluate the effects of single nucleotide polymorphisms (SNPs) of long intergenic non-coding RNAs (lincRNAs) on ESCC susceptibility in Chinese populations. We scoured exons of lincRNAs located in ESCC susceptibility loci for all probable functional SNPs. These 52 SNPs were opted for and genotyped in 1493 ESCC patients and 1553 cancer-free controls from eastern and southern Chinese populations, and their associations with the risk for ESCC were estimated using logistic regression. Functional relevance was further examined by biochemical assays. Significant differences were found between patients and controls in the genotype frequencies for the rs11752942A>G site in the lincRNA-uc003opf.1 exon. Compared with the rs11752942AA genotype, AG and GG genotypes had a significantly reduced risk of ESCC (adjusted odds ratio = 0.73; 95% confidence interval = 0.63-0.84). Biochemical analysis demonstrated that, when compared with the A allele, the rs11752942G allele could markedly attenuate the level of lincRNA-uc003opf.1 both in vivo and in vitro by binding micro-RNA-149*, thereby affecting cell proliferation and tumor growth. These findings indicated that functional polymorphism rs11752942A>G in lincRNA-uc003opf.1 exon might be a genetic modifier for the development of ESCC.


Wave refraction in negative-index media: always positive and very inhomogeneous.
We present the first treatment of the refraction of physical electromagnetic waves in newly developed negative index media (NIM), also known as left-handed media (LHM). The NIM dispersion relation implies that group fronts refract positively even when phase fronts refract negatively. This difference results in rapidly dispersing, very inhomogeneous waves. In fact, causality and finite signal speed always prevent negative wave signal (not phase) refraction. Earlier interpretations of phase refraction as "negative light refraction" and "light focusing by plane slabs" are therefore incorrect, and published NIM experiments can be explained without invoking negative signal refraction.


Exciton Localization on Ru-Based Photosensitizers Induced by Binding to Lipid Membranes.
The characterization of electronic properties of metal complexes embedded in membrane environments is of paramount importance to develop efficient photosensitizers in optogenetic applications. Molecular dynamics and QM/MM simulations together with quantitative wave function analysis reveal a directional electronic redistribution of the exciton formed upon excitation of [Ru(bpy)2(bpy-C17)]2+ when going from water to a lipid bilayer, despite the fact that the media influence neither the metal-to-ligand charge-transfer character nor the excitation energy of the absorption spectra. When the photosensitizer is embedded into the DOPC lipid membrane, exciton population is mainly located in the bypyridyl sites proximal to the positively charged surface of the bilayer due to electrostatic interactions. This behavior shows that the electronic structure of metal complexes can be controlled through the binding to external species, underscoring the crucial role of the environment in directing the electronic flow upon excitation and thus helping rational tuning of optogenetic agents.


Gene expression analysis reveals matrilysin as a key regulator of pulmonary fibrosis in mice and humans.
Pulmonary fibrosis is a progressive and largely untreatable group of disorders that affects up to 100,000 people on any given day in the United States. To elucidate the molecular mechanisms that lead to end-stage human pulmonary fibrosis we analyzed samples from patients with histologically proven pulmonary fibrosis (usual interstitial pneumonia) by using oligonucleotide microarrays. Gene expression patterns clearly distinguished normal from fibrotic lungs. Many of the genes that were significantly increased in fibrotic lungs encoded proteins associated with extracellular matrix formation and degradation and proteins expressed in smooth muscle. Using a combined set of scoring systems we determined that matrilysin (matrix metalloproteinase 7), a metalloprotease not previously associated with pulmonary fibrosis, was the most informative increased gene in our data set. Immunohistochemisry demonstrated increased expression of matrilysin protein in fibrotic lungs. Furthermore, matrilysin knockout mice were dramatically protected from pulmonary fibrosis in response to intratracheal bleomycin. Our results identify matrilysin as a mediator of pulmonary fibrosis and a potential therapeutic target. They also illustrate the power of global gene expression analysis of human tissue samples to identify molecular pathways involved in clinical disease.


Preparation of a Highly Conductive Seed Layer for Calcium Sensor Fabrication with Enhanced Sensing Performance.
The seed layer plays a crucial role in achieving high electrical conductivity and ensuring higher performance of devices. In this study, we report fabrication of a solution-gated field-effect transistor (FET) sensor based on zinc oxide nanorods (ZnO NRs) modified iron oxide nanoparticles (α-Fe2O3 NPs) grown on a highly conductive sandwich-like seed layer (ZnO seed layer/Ag nanowires/ZnO seed layer). The sandwich-like seed layer and ZnO NRs modification with α-Fe2O3 NPs provide excellent conductivity and prevent possible ZnO NRs surface damage from low pH enzyme immobilization, respectively. The highly conductive solution-gated FET sensor employed the calmodulin (CaM) immobilization on the surface of α-Fe2O3-ZnO NRs for selective detection of calcium ions (Ca2+). The solution-gated FET sensor exhibited a substantial change in conductance upon introduction of different concentrations of Ca2+ and showed high sensitivity (416.8 μA cm-2 mM-1) and wide linear range (0.01-3.0 mM). In addition, the total Ca2+ concentration in water and serum samples was also measured. Compared to the analytically obtained data, our sensor was found to measure Ca2+ in the water and serum samples accurately, suggesting a potential alternative for Ca2+ determination in water and serum samples, specifically used for drinking/irrigation and clinical analysis.


Overexpression of hyaluronan synthase-2 reduces the tumorigenic potential of glioma cells lacking hyaluronidase activity.
The interactions of CD44 with hyaluronan are thought to be crucial for tumor cell attachment to the extracellular matrix, migration, and invasion. For migration to occur, however, the interactions between hyaluronan and cell surface receptors need to be transient. Hyaluronidases may facilitate the degradation of hyaluronan bound to the cell surface and thus reduce the interactions of the cells with the matrix, whereas the overproduction of hyaluronan in the absence of hyaluronidase activity may prevent cells from proliferating or invading normal surrounding tissue. We analyzed the effects in vitro and in vivo of hyaluronan synthase-2 (HAS2) overexpression on a murine glioma cell line that is deficient in hyaluronidase activity. In addition, we evaluated the expression levels of HAS and hyaluronidase genes in human glioma cell lines and in glioma specimens. Increased hyaluronan synthesis had no effect on the in vitro proliferation of the cells but diminished their in vivo growth rate. Several human glioma cell lines were found to overexpress hyaluronan synthases, but they did so in conjunction with hyaluronidase Hyal2 and MGEA5 expression. Similarly, all glioblastomas multiforme expressed hyaluronidases MGEA5 and Hyal2. The data suggest that an increased synthesis of hyaluronan by astrocytoma cells is only promoting tumor cell growth in vivo if the cells express hyaluronidases as well.


Intravenous immune globulin for the prevention of bacterial infections in children with symptomatic human immunodeficiency virus infection.
Serious recurrent bacterial infections are a major cause of morbidity and mortality in children infected with the human immunodeficiency virus (HIV). Because intravenous immune globulin has been shown to prevent bacterial infection in patients with primary immunodeficiency and in uncontrolled studies of HIV-infected children, we undertook a multicenter study of its safety and efficacy in children with symptomatic HIV infection. In a double-blind trial, 372 HIV-infected children (mean age, 40 months) with clinical or immunologic evidence of HIV disease were randomly assigned to receive either intravenous immune globulin (400 mg per kilogram of body weight) or placebo (0.1 percent albumin) every 28 days. The children were stratified into two groups according to CD4+ lymphocyte count at entry into the study and the clinical classification of the Centers for Disease Control. The median length of follow-up was 17 months. For children in either group with CD4+ counts greater than or equal to 0.2 x 10(9) per liter (greater than or equal to 200 per cubic millimeter) at entry, treatment with intravenous immune globulin significantly increased the time free from serious infection; estimated infection-free rates after 24 months were 67 percent for children receiving immune globulin as compared with 48 percent for those receiving placebo (P = 0.01). In addition, immune globulin was associated with an overall reduction in the number of both serious and minor bacterial infections (relative risk, 0.68; P = 0.01) and in the number of hospitalizations for acute care (relative risk, 0.65; P = 0.03). No such benefits were seen for children with CD4+ counts below 0.2 x 10(9) per liter at entry. For group 1 overall, there was a trend toward a difference in serious bacterial infection between immune globulin and placebo (24-month infection-free survival, 31 percent for intravenous immune globulin vs. 25 percent for placebo; P = 0.10). For group 2, the estimates of survival without serious infection were 73 percent with intravenous immune globulin as compared with 53 percent with placebo (P = 0.04). There was no effect of treatment on mortality for any group or CD4+ count at entry. Adverse reactions, noted for less than 1 percent of infusions, were minor. In symptomatic HIV-infected children the prophylactic use of intravenous immune globulin, is safe, and it significantly increases the time free from serious bacterial infections for those entering treatment with CD4+ lymphocyte counts greater than or equal to 0.2 x 10(9) per liter.


The influence of hormonal replacement and growth hormone treatment on the lipids in Turner syndrome.
Women with Turner syndrome (TS) have a risk of developing cardiovascular diseases. We assessed the lipid and carbohydrate metabolism in TS-women in the context of current hormone replacement therapy (HRT) and growth hormone (GH) treatment during childhood. The information were collected from medical documentation and anamnesis of 165 TS-women (24.9 ± 7.7 yr) between 1995 and 2011. The patients underwent a pituitary-gonadal axis assessment together with measurements of total cholesterol (TC), high- (HDL) and low- (LDL) density lipoproteins, triglycerides (TG), and glucose levels. Only 58% of women were using HRT. No differences were found in the levels of the lipid components and glucose in women who were undergoing HRT compared to those without it. Compared to TS-women without (n = 113), prior GH treatment in 34 TS-women positively influenced the lipid parameters: TC 5.0 ± 1.1 versus 4.6 ± 0.9 mmol/l (p = 0.03), HDL 1.5 ± 0.5 versus 1.4 ± 0.4 mmol/l (p > 0.05), LDL 3.3 ± 0.9 versus 2.9 ± 0.7 mmol/l (p = 0.03), and TG 1.1 ± 0.6 versus 0.8 ± 0.3 g/l (p = 0.009), respectively. (1) HRT does not affect lipid metabolism in TS-women. (2) The use of GH in TS-children favorably influences their lipid profile in adulthood.


Mechanism of action of IFN-beta in the treatment of multiple sclerosis: a special reference to CD73 and adenosine.
IFN-beta treatment reduces the relapse rate in multiple sclerosis (MS), but the exact mechanism of action of the drug has remained elusive. CD73 (ecto-5'-nucleotidase) is an ectoenzyme, which produces adenosine from adenosine monophosphate (AMP) precursor by enzymatic dephosphorylation. AMP is known to be abundantly present at sites of inflammation, and more importantly adenosine, the product of CD73, is known to possess both anti-inflammatory and neuroprotective activity. Our preliminary work has shown that IFN-beta increases the expression of ecto-5'-nucleotidase on endothelial cells (ECs) both in vitro and after systemic treatment of MS patients in vivo. In the majority of MS patients also an increase in the soluble serum CD73 was noted after IFN-beta treatment. Importantly, this correlated with the clinical outcome. CD73 expression on central nervous system (CNS) microvasculature was confirmed with stainings of frozen tissue sections of MS brain samples taken at autopsy. Adenosine, a known neuroprotective agent, might contribute to the beneficial effects of IFN-beta on MS.


EIPL (extensive intraoperative peritoneal lavage) therapy significantly reduces peritoneal recurrence after pancreatectomy in patients with pancreatic cancer.
One standing problem in the therapy for pancreatic cancer is peritoneal recurrence after pancreatectomy. We have established EIPL (extensive intra-operative peritoneal lavage) therapy as a very simple and non-aggressive prophylactic treatment for peritoneal dissemination of gastric cancer patients with peritoneal free cancer cells. In this study, the impact of EIPL therapy in preventing peritoneal recurrence in patients with pancreatic cancer after a curative surgical operation is analyzed. This study was based on 39 consecutive patients with invasive ductal adenocarcinoma of the pancreas who underwent curative surgical treatment. The patients were divided into two groups: the non-EIPL group, patients without EIPL therapy (from 1995 to 2000) (n=24); and the EIPL group, patients with EIPL therapy (from 2001 to 2003) (n=15). Clinicopathological findings, recurrence patterns and cancer-free survival were compared between the two groups. In addition, to identify free cancer cells during the operation, both carcinoembryonic antigen (CEA)- and cytokeratin 20 (CK20)-specific intraoperative real-time RT-PCR was performed immediately after laparotomy, lymph node dissection and removal of the pancreas in six patients with curative pancreatic cancer. Peritoneal, hepatic, lymphatic, local, and extra-abdominal recurrence rates in the EIPL and non-EIPL groups were 6.7, 40.0, 26.7, 13.3 and 13.3%, and 45.8, 50.0, 20.8, 29.2 and 20.8%, respectively. Among these recurrence patterns, the peritoneal recurrence rate of the EIPL group was significantly lower than that of the non-EIPL (P=0.013). Real-time RT-PCR analysis identified intra-peritoneal free cancer cells in 0, 33 and 67% of patients immediately after laparotomy, lymph node dissection and removal of the pancreatic cancer, respectively. After EIPL therapy, however, no cancer cell was detected in the peritoneal cavity. EIPL therapy was the independent negative risk factor for peritoneal recurrence (P=0.044), and the cancer-free 2-year survival of the EIPL group showed improvement compared with that of the non-EIPL group and was near statistical significance (P=0.097). EIPL therapy significantly prevented peritoneal recurrence after curative operation of pancreatic cancer, and it may contribute to reducing the mortality rate of this aggressive disease. Continued investigation of this promising treatment regimen is warranted.


Two GDP-mannose transporters contribute to hyphal form and cell wall integrity in Aspergillus nidulans.
In order to identify novel genes affecting cell wall integrity, we have generated mutant strains of the filamentous fungus Aspergillus nidulans that show hypersensitivity to the chitin-binding agent Calcofluor White (CFW). Affected loci are designated cal loci. The phenotype of one of these alleles, calI11, also includes shortened hyphal compartments and increased density of branching in the absence of CFW, as well as reduced staining of cell walls by the lectin FITC-Concanavalin A (ConA), which has strong binding affinity for mannosyl residues. We have identified two A. nidulans genes (AN8848.3 and AN9298.3, designated gmtA and gmtB, respectively) that complement all aspects of the phenotype. Both genes show strong sequence similarity to GDP-mannose transporters (GMTs) of Saccharomyces and other yeasts. Sequencing of gmtA from the calI11 mutant strain reveals a G to C mutation at position 943, resulting in a predicted alanine to proline substitution at amino acid position 315 within a region that is highly conserved among other fungi. No mutations were observed in the mutant strain's allele of gmtB. Meiotic mapping demonstrated a recombination frequency of under 1 % between the calI locus and the phenA locus (located approximately 9.5 kb from AN8848.3), confirming that gmtA and calI are identical. A GmtA-GFP chimera exhibits a punctate distribution pattern, consistent with that shown by putative Golgi markers in A. nidulans. However, this distribution did not overlap with that of the putative Golgi equivalent marker CopA-monomeric red fluorescent protein (mRFP), which may indicate that the physically separated Golgi-equivalent organelles of A. nidulans represent physiologically distinct counterparts of the stacked cisternae of plants and animals. These findings demonstrate that gmtA and gmtB play roles in cell wall metabolism in A. nidulans similar to those previously reported for GMTs in yeasts.


Management of comedonal acne vulgaris with fixed-combination topical therapy.
Acne vulgaris (acne) is the most common skin disease we see in dermatology practice. Clinically, it is characterized by a combination of open and closed comedones (formally referred to as noninflammatory lesions) and inflammatory papules and pustules. Comedonal acne is more typical in young adolescents, but can occur in combination with inflammatory papules and pustules at any time. Topical retinoids have long been advocated for the treatment of comedonal acne. Given the increasing recognition of the inflammatory nature of acne and the synergistic benefits seen with fixed combinations we review the latest clinical data to provide guidance on optimal management of comedonal acne. An English language literature search of Medline, EMBASE, and the Web of Science using key terms (acne, comedonal, noninflammatory, clinical trials) was conducted, and relevant articles reviewed. Comparative data is sparse, but we show the importance of fixed combinations with and without retinoids, where treatment benefits are comparable. Adapalene 0.1%-benzoyl peroxide 2.5% gel has been shown to be comparable to clindamycin 1%-benzoyl peroxide 5% gel, and adapalene 0.3%-benzoyl peroxide 2.5% gel. A meta-analysis suggested that clindamycin 1.2%-benzoyl peroxide 2.5% gel was more effective than clindamycin-benzoyl peroxide 5% gel in noninflammatory lesions, and two equivalent clinical programs suggest additional benefits of higher doses of benzoyl peroxide (3.75% vs 2.5%) in this fixed combination. Clindamycin 1.2%-benzoyl peroxide 3.75% gel may afford similar benefits to adapalene 0.3%-benzoyl peroxide 2.5% gel in this sometimes difficult to treat patient population.


[Antenatal diagnosis of placenta accreta].
Placenta accreta is a potentially life-threatening obstetric condition. Diagnosis of placenta accreta before delivery allows multidisciplinary planning in an attempt to minimize potential maternal or neonatal morbidity and mortality The diagnosis is usually established by 2D, 3D Ultrasonography and Color Doppler ultrasonography and occasionally supplemented by Magnetic Resonance Imaging.


[Functional hyperandrogenism--classification, etiology, diagnosis and therapy].
The classification of functional hyperandrogenism (FHA) presented in this paper is based on well known clinical experience supported by recent data of molecular biology. Functional hyperandrogenism is composed of various organ system-specific entities with consequently differential diagnostic and therapeutic strategies. The term polycystic ovary syndromes (PCOS) is misleading and should be replaced by adequate descriptions. Inspite of intense discussions and progress in molecular biology are with the exception of the here described FHA III-group the etiological consequences unresolved in terms of diagnostic and therapeutic procedures. Based on recent findings on the human genome genetic screening methods (Microarrays) may be available in the near future to allow a better understanding of the underlying pathophysiology.


Adipose tissue lipolysis revisited (again!): lactate involvement in insulin antilipolytic action.
Lactate is produced from glucose by adipose tissue. In this issue of Cell Metabolism, Ahmed et al. (2010) show that the metabolite is involved in the antilipolytic effect of insulin, providing a new link between fat and carbohydrate metabolism.


Organization and promoter analysis of the zebrafish (Danio rerio) interferon gene.
Interferon plays important roles in confronting viral infections as the first line of defense. For the purpose of understanding the molecular mechanism which controls transcription of the interferon gene, we cloned and sequenced the interferon promoter region of the zebrafish interferon gene and characterized its activity using firefly luciferase transient transfection expression assays. Different fragments of the zebrafish interferon 5'-flanking region were transfected into ZFL cells. In these cell lines, maximum promoter activity was located in 2.2 kb of the zebrafish interferon 5' flanking region of the ZFL cell line. In this study, we investigated whether the viral replicative intermediate double-stranded RNA (herein we used synthetic polyinosinic-polycytidylic acid [poly(I):poly(C)] modifies the effects of interferon on gene expression. For this purpose, all zebrafish interferon promoter fragments were treated with either 1, 10, or 100 microg/ml poly(I):poly(C). The results showed that after treatment with 10 microg/ml poly(I):poly(C), high promoter activity was observed in the -2.2-kb interferon promoter fragment. Several putative transcription factors were shown in the promoter region, including IRF-1, C/EBP, NFkappaB, and GATA-1. Further study of the in vivo expression of the interferon promoter during development was carried out in transgenic zebrafish expressing an interferon promoter-driven green fluorescent protein (GFP) encoding the GFP cDNA transgene. Morphological studies of transgenic zebrafish indicated that the interferon promoter-driven GFP transcripts appeared in the yolk, head, and lymphoid organs. These results indicate that the interferon promoter is active in a tissue-specific manner, and suggest that the interferon promoter plays an important role in virus resistance during teleost growth.


Characterizing rhizome bud dormancy in Polygonatum kingianum: Development of novel chill models and determination of dormancy release mechanisms by weighted correlation network analysis.
This study was conducted to explore specific chill models and the mechanisms underlying rhizome bud dormancy break in Polygonatum kingianum. Rhizome buds were subjected to various chilling temperatures for different duration and then transferred to warm conditions for germination and subsequent evaluation of their response to temperature and chilling requirements. A CUkingianum model was constructed to describe the contribution of low temperature to the chill unit, and it was suggested that 2.97°C was the optimum temperature and that 11.54°C was the upper limit for bud release. The CASkingianum model showed the relationship between chilling accumulation and sprouting percentage; therefore, rhizome bud development could be predicted through the model. Weighted correlation network analysis (WGCNA) of transcriptomic data of endo-, eco- and nondormant rhizome buds generated 33 gene modules, 6 of which were significantly related to bud sprouting percentage. In addition, 7 significantly matched transcription factors (TFs) were identified from the promoters of 17 "real" hub genes, and DAG2 was the best matched TF that bound to AAAG element to regulate gene expression. The current study is valuable for developing a highly efficient strategy for seedling cultivation and provides strong candidates for key genes related to rhizome bud dormancy in P. kingianum.


The immunogenicity and reactogenicity of four COVID-19 booster vaccinations against SARS-CoV-2 variants following CoronaVac or ChAdOx1 nCoV-19 primary series.
The appropriate COVID-19 booster vaccine following inactivated or adenoviral vector COVID-19 vaccination is unclear. To investigate the immunogenicity of four COVID-19 booster vaccines. We prospectively enrolled healthy adults who received a two-dose CoronaVac or ChAdOx1 8-12 weeks earlier and allocated them to receive one of the following booster vaccine: inactivated (BBIBP-CorV), ChAdOx1 or mRNA (BNT162b2 at full [30 μg] and half [15 μg] dose) vaccines. We determined the reactogenicity and the humoral (anti-receptor binding domain IgG (anti-RBD-IgG), neutralizing antibodies (nAb) against Delta, Beta and Omicron variants) and cellular immunity measuring by interferon gamma (IFN-γ) responses post-booster. AR patients. Among the 352 participants (179 CoronaVac and 173 ChAdOx1 participants), 285 (81%) were female, and median age was 39 (IQR: 31-47) years. Two weeks post-booster, both 30 μg- and 15 μg- BNT162b2 induced the highest anti-RBD IgG concentration (BAU/mL); Coronavac-prime: 30 μg-BNT162b2, 5152.2 (95%CI 4491.7-5909.8); 15 μg-BNT162b2, 3981.1 (3397.2-4665.4); ChAdOx1, 1358.0 (1141.8-1615.1); BBIBP-CorV, 154.6 (92.11-259.47); ChAdOx1-prime: 30 μg-BNT162b2, 2363.8 (2005.6-2786.1; 15 μg-BNT162b2, 1961.9 (1624.6-2369.1); ChAdOx1, 246.4 (199.6-304.2); BBIBP-CorV, 128.1 (93.5-175.4). Similarly, both 30 μg- and 15 μg- BNT162b2 boosting induced the highest nAb titers against Beta, Delta and Omicron BA.1 variants and highest T-cell response at 2 weeks after boosting. While all BNT162b2 or heterologous ChAdOx1-boosted participants had nAb against Omicron, these were < 50% for BBIBP-CorV and 75% for homologous ChAdOx1-boosted participants. There was significant decrease in nAb ( > 4-fold) at 16-20 weeks post booster for all groups. Heterologous boosting with BNT162b2 following CoronaVac or ChAdOx1 primary series is most immunogenic. Additional studies are needed to verify the clinical efficacy and persistence of immunity following half-dose BNT162b2.


A phase 1 healthy male volunteer single escalating dose study of the pharmacokinetics and pharmacodynamics of risdiplam (RG7916, RO7034067), a SMN2 splicing modifier.
Risdiplam (RG7916, RO7034067) is an orally administered, centrally and peripherally distributed, survival of motor neuron 2 (SMN2) mRNA splicing modifier for the treatment of spinal muscular atrophy (SMA). The objectives of this entry-into-human study were to assess the safety, tolerability, pharmacokinetics (PK) and pharmacodynamics of risdiplam, and the effect of the strong CYP3A inhibitor itraconazole on the PK of risdiplam in healthy male volunteers. Part 1 had a randomized, double-blind, adaptive design with 25 subjects receiving single ascending oral doses of risdiplam (ranging from 0.6-18.0 mg, n = 18) or placebo (n = 7). A Bayesian framework was applied to estimate risdiplam's effect on SMN2 mRNA. The effect of multiple doses of itraconazole on the PK of risdiplam was also assessed using a two-period cross-over design (n = 8). Risdiplam in the fasted or fed state was well tolerated. Risdiplam exhibited linear PK over the dose range with a multi-phasic decline with a mean terminal half-life of 40-69 h. Food had no relevant effect, and itraconazole had only a minor effect on plasma PK indicating a low fraction of risdiplam metabolized by CYP3A. The highest tested dose of 18.0 mg risdiplam led to approximately 41% (95% confidence interval 27-55%) of the estimated maximum increase in SMN2 mRNA. Risdiplam was well tolerated and proof of mechanism was demonstrated by the intended shift in SMN2 splicing towards full-length SMN2 mRNA. Based on these data, Phase 2/3 studies of risdiplam in patients with SMA are now ongoing.


5,7-Dihydroxytryptamine induced changes in the postnatal development of central 5-hydroxytryptamine neurons.
Systemic administration of the neurotoxic compound 5,7-dihydroxytryptamine (5,7-HT) to newborn rats led acutely (within 1--2 h) to a marked reduction of the in vitro uptake of [3H]5-hydroxytryptamine (5-HT) in homogenates from the cerebral cortex (75% decrease) and the pons-medulla (60% decrease). When 5,7-HT was administered postnatally we found that between days 5 an7 resistance developed in the cerebral cortex towards the 5,7-HT induced reduction in 3H-5-HT uptake. This was most likely the result of the postnatal development of the blood-brain barrier. These results show that 5,7-HT can pass the blood-brain barrier in the neonate stage and enter the brain to exert its well-known neurotoxic action on 5-HT neurons. The [3H]5-HT uptake in the cerebral cortex was reduced quantitatively to the same extent up to the 28th postnatal day, after which time a moderate recovery took place. Endogenous 5-HT was reduced by 40% in the cerebral cortex when measured in adult animals. In the pons-medulla there was a rapid recovery of the [3H]5-HT uptake during the first week after the 5,7-HT treatment and on the 14th postnatal day the increase was as much as 75% compared with the control. Endogenous 5-HT and [3H]5-HT uptake was increased by 40--50% when the analysis was performed 2 months after the 5,7-HT treatment. Studies of [3H]noradrenaline (NA) uptake after 5,7-HT administration at birth showed that this treatment similarly affected the NA neurons, though to a lesser extent. The effects on the NA neurons could be abolished by pretreatment with the "membrane pump" blocker desipramine, leaving the action of 5,7-HT on 5-HT neurons almost unaffected. Analysis of the [3H]5-HT uptake kinetics in the pons-medulla showed that the 5,7-HT treatment did not affect the Km while the Vmax was increased. It is concluded that neonatal 5,7-HT treatment produces a marked 5-HT denervation of the cerebral cortex, while there is a stimulated postnatal outgrowth of 5-HT nerve terminals in the pons-medulla, after an initial partial damage of the neurons.


Switch effect of the nonquantized intrinsic spin Hall conductivity in monolayered monoclinic transition metal dichalcogenides.
First-principles calculations have been performed to study the intrinsic spin Hall effect (SHE) and its behavior under vertical electric field in monoclinic transition metal dichalcogenide monolayers (1T'-MX2 with M = Mo, W and X = S, Se, Te). We find that the pristine systems exhibit nonquantized intrinsic spin Hall conductivity (SHC) due to the unconserved spin around the direct band gaps though they have nontrivial band topology. The unconserved spin is attributed to the band crossings at Fermi levels for systems without spin-orbit coupling and the distinct composition of the band states around the crossings. Despite the nonquantization of SHC, calculations with the hybrid functional predict SHC approaching the quantized value in W based systems, especially 1T'-WTe2, which has been realized in experiments. More interesting, a sharp drop of SHC to almost zero in semiconducting systems induced by vertical electric field is observed at the topological phase transition point, suggesting that such systems exhibit a strong switch effect of SHC. In contrast, the switch effect is weak in semi-metallic systems, where the SHC decreases almost continuously with increasing field strength for the chemical potential around the Fermi levels. Our findings suggest potential applications of the pristine 1T'-MX2 and those under vertical electric field in spintronics devices by utilizing the intrinsic SHE of their bulk states.


Does the Gillet test assess sacroiliac motion or asymmetric one-legged stance strategies?
The purpose of this study was to quantify the extent to which apparent movements of the posterior superior iliac spine and sacral base areas Gillet sacroiliac motion testing were related to (a) degree of hip flexion and (b) the examiner's palpatory pressure. A preliminary exploratory study quantified relative PSIS/S2 displacements in 10 sacroiliac joints among 5 asymptomatic subjects at 10° increments of hip flexion from 0-90°. A comprehensive follow-up asymptomatic study quantified PSIS/S2 displacements at 0° vs. 30° vs. 90° hip flexion, and for light vs. firm pressure at 30° hip flexion. Displacements measured in pixels on digital photographs were transformed to mm. Mean differences for the various test conditions were evaluated for statistical significance using paired t-testing and Wilcoxon signed rank test. With light pressure, the left PSIS moved caudal for hip flexion ≤30° during right-legged stance, whereas the right PSIS moved cephalad relative to the sacral base. For hip flexion =90°, both PSISs moved cephalad. The use of firm palpatory pressure abolished the initial caudal movement of the left PSIS, as well as differences in the amount of cephalad PSIS movement at 30° vs. 90° hip flexion. The results are consistent with there being left-right differences in gluteus medius and biceps femoris activation among asymptomatic individuals that result in different balancing strategies during one-legged stance. This may create the appearance of relative PSIS/SB displacement, even though the results of Gillet testing can be wholly or partially explained by pelvic obliquity owing to muscle function asymmetry. This study questions the validity of the upright Gillet test for sacroiliac motion.


Application of image spectrometer to in situ infrared broadband optical monitoring for thin film deposition.
A path-folded infrared image spectrometer with five sub-gratings and five linear-array detectors was applied to a broadband optical monitoring (BOM) system for thin film deposition. Through in situ BOM, we can simultaneously acquire the thickness and refractive index of each layer in real time by fitting the measured spectra, and modify the deposition parameters during deposition process according to the fitting results. An effective data processing method was proposed and applied in the BOM process, and it shortened the data processing time and improved the monitoring efficiency greatly. For demonstration, a narrow band-pass filter (NBF) at 1540 nm with ~10 nm full width at half-maximum (FWHM) had been manufactured using the developed BOM system, and the results showed that this BOM method was satisfying for monitoring deposition of thin film devices.


Early growth response protein 1 promotes restenosis by upregulating intercellular adhesion molecule-1 in vein graft.
To verify the relationship between Egr-1 and vein graft restenosis and investigate the related mechanisms. Mouse vein graft models were established in Egr-1 knockout (KO) and wild-type (WT) mice. The vein grafts in the mice were taken for pathological examination and immunohistochemical analysis. The endothelial cells (ECs) were stimulated by using a computer-controlled cyclic stress unit. BrdU staining and PCR were used to detect ECs proliferation activity and Egr-1 and ICAM-1 mRNA expression, respectively. Western-blot analysis was also used to detect expression of Egr-1 and intercellular adhesion molecule-1 (ICAM-1) proteins. The lumens of vein grafts in Egr-1 KO mice were wider than in WT mice. ECs proliferation after mechanical stretch stimulation was suppressed by Egr-1 knockout (P < 0.05). Both in vein grafts and ECs from WT mice after mechanical stretch stimulation, mRNA expression and protein of Egr-1 and ICAM-1 showed increases (P < 0.05). However, ICAM-1 expression was significantly suppressed in ECs from Egr-1 knockout mice (P < 0.05). Egr-1 may promote ECs proliferation and result in vein graft restenosis by upregulating the expression of ICAM-1. As a key factor of vein graft restenosis, it could be a target for the prevention of restenosis after CABG surgery.


Overexpression of the Hsa21 Transcription Factor RUNX1 Modulates the Extracellular Matrix in Trisomy 21 Cells.
Down syndrome is a neurodevelopmental disorder frequently characterized by other developmental defects, such as congenital heart disease. Analysis of gene expression profiles of hearts from trisomic fetuses have shown upregulation of extracellular matrix (ECM) genes. The aim of this work was to identify genes on chromosome 21 potentially responsible for the upregulation of ECM genes and to pinpoint any functional consequences of this upregulation. By gene set enrichment analysis of public data sets, we identified the transcription factor RUNX1, which maps to chromosome 21, as a possible candidate for regulation of ECM genes. We assessed that approximately 80% of ECM genes overexpressed in trisomic hearts have consensus sequences for RUNX1 in their promoters. We found that in human fetal fibroblasts with chromosome 21 trisomy there is increased expression of both RUNX1 and several ECM genes, whether located on chromosome 21 or not. SiRNA silencing of RUNX1 reduced the expression of 11 of the 14 ECM genes analyzed. In addition, collagen IV, an ECM protein secreted in high concentrations in the culture media of trisomic fibroblasts, was modulated by RUNX1 silencing. Attenuated expression of RUNX1 increased the migratory capacity of trisomic fibroblasts, which are characterized by a reduced migratory capacity compared to euploid controls.


Measurement and analysis of diode laser wave fronts.
The wave fronts of several types of diode laser are investigated by means of Mach-Zehnder and phase difference amplification interferometers. The astigmatic aberration tolerance of each diode laser is estimated using aberration theory to determine if the lasers are suitable as light sources for precise optical systems, such as laser beam printers and optical disks.


The nucleolar structure and the activity of NopA100, a nucleolin-like protein, during the cell cycle in proliferating plant cells.
For the purpose of gaining knowledge of the relationships between cell proliferation and ribosome biogenesis, as two fundamental mutually interconnected cellular processes, studies were performed on cell populations synchronized in their cell-cycle progression by treatment with hydroxyurea, followed by sampling at different times after its removal. A structural rearrangement of the nucleolus was observed throughout the interphase, along with changes in the relative amounts of different nucleolar subcomponents. A structural model of nucleolar organization was associated with each interphase period. Throughout interphase, the nucleolin-like protein, NopA100, was immunodetected in the dense fibrillar component of the nucleolus, preferentially near fibrillar centers and its levels were shown to increase from G1 to G2. A western blotting analysis of soluble nuclear protein extracts with anti-NopA100 antibody resulted in the intense labeling of a 100-kDa band, but also of a series of proteins related to it, suggesting that NopA100 undergoes a physiological process of proteolytic maturation, similar to that described for mammalian nucleolin, but not reported in other biological model systems. Physiological proteolysis of NopA100, related to cell-cycle progression, was confirmed after the nuclei extracted from synchronized cells were treated with the protease inhibitor, leupeptin, which resulted in an increase of the 100-kDa band at the expenses of the decrease of some other bands, according to the cell-cycle stages. We therefore conclude that there is a relationship between the increase in nucleolar activity, cell-cycle progression, nucleolar structure, the activity of NopA100, and the proteolysis of this nucleolin-like protein.


Active surveillance of renal masses: an analysis of growth kinetics and clinical outcomes stratified by radiological characteristics at diagnosis.
To determine the growth rate of renal masses (RMs) under active surveillance (AS), and to describe the clinical outcome of AS patients. We conducted a retrospective review of an AS database to obtain demographics, radiological and pathologic characteristics and RM size of patients. RMs were followed at 6-12 month intervals for ≥1 year with computed tomography (CT), magnetic resonance imaging (MRI), or renal ultrasound. Kaplan-Meier analysis determined the annual likelihood of intervention. RMs were divided into 3 radiographic subcategories (solid, cystic, and angiomyolipoma). A linear regression model determined RM growth rates. 131 RMs in 114 patients were included. Median age, Charlson Comorbidity Index score and mean follow-up were 69.1 years, 4.0 and 4.2±2.6 years, respectively. Maximal tumor diameter (MTD) at diagnosis was 2.1 ± 1.3 cm. 49 RMs exhibited negative or zero net growth. Mean MTD growth rate for all RMs was 0.72±3.2 (95% CI: 0.16-1.28) mm/year. When stratified by MTD at diagnosis, mean RM growth rates were 0.84, 0.84, 0.44, 0.74 and 0.71 mm/year for RMs ≤1 cm, 1-≤2cm, 2-≤ 3cm, 3-≤ 4cm and ≥4cm, respectively (p≤0.01). The 5 and 10-year freedom from intervention rates were 93.1% and 88.5%, respectively. There was a single case of suspected metastases, but no deaths related to kidney cancer. RMs under AS grew slowly, and had a low incidence of requiring surgical intervention and progression. Solid enhancing masses grew slowly, and were more likely to trigger intervention. AS should be considered for selected patients with small RMs.


End-effector lower limb robot-assisted gait training effects in subacute stroke patients: A randomized controlled pilot trial.
This pilot study investigated end-effector lower limb rehabilitation robot training effects in subacute stroke patients. Forty-nine stroke patients were randomly assigned to 2 treatment groups: a 30-minute end-effector lower limb rehabilitation robot training plus 1.5-hour conventional physiotherapy (robot group; n = 26), or a 2-hour conventional physiotherapy (control group; n = 23). All patients received 5 treatments weekly for 4 weeks. The functional ambulatory category was the primary outcome and the motricity index, Fugl Meyer assessment-lower extremity, rivermead mobility index, 10 meter walk test, Berg balance scale, and modified Barthel index were secondary outcomes. All outcome measures significantly improved in both groups after training (P > .05). The robot group improved more in FAC than the control group (P = .005). Compared with conventional physiotherapy alone, end-effector lower limb robot-assisted gait training with conventional physiotherapy improved subacute stroke patients walking ability.


HIV Treatment with the Two-Drug Regimen Dolutegravir Plus Lamivudine in Real-world Clinical Practice: A Systematic Literature Review.
The two-drug regimen dolutegravir plus lamivudine demonstrated durable efficacy for up to 3 years in phase III studies and a high barrier to resistance in treatment-naive and virologically suppressed people with HIV (PWH). This systematic literature review summarizes real-world evidence evaluating effectiveness and safety of dolutegravir plus lamivudine. We searched Ovid MEDLINE®, Embase®, PubMed, Cochrane library, and relevant international conference proceedings from 2013 to 2020. Qualitative synthesis of virologic suppression at Week 48, treatment-emergent resistance, discontinuation rates, and comorbidities was undertaken, with no statistical analyses conducted. Linked publications and potential for duplication in reporting of outcomes for cohorts and populations were identified, and the publication reporting the highest number of PWH receiving dolutegravir plus lamivudine was included in the analysis. Thirty-four studies reporting on cohorts of PWH not suspected to be linked or to include duplicate data receiving dolutegravir plus lamivudine were identified (N = 5017). Of 3744 virologically suppressed PWH who switched to dolutegravir plus lamivudine, 603 (16%) reported history of virologic failure. Nineteen studies included effectiveness data (n = 3558), four of which included data from treatment-naive PWH (n = 69). In studies with > 100 PWH, high rates of virologic suppression (Week 48, 97-100%) were maintained with dolutegravir plus lamivudine, with low rates of virologic failure (0-3.3 per 100 person-years of follow-up); one instance of emergent integrase strand transfer inhibitor resistance was reported in a complex treatment-experienced individual. Rates of discontinuation due to adverse events were low and consistent with previously observed trial data. Dolutegravir plus lamivudine minimally impacted renal function and had minimal impact on or improved lipid profiles and bone mineral density. This systematic review demonstrates that effectiveness and safety of dolutegravir plus lamivudine in clinical practice support data from randomized controlled trials with regard to high rates of virologic response, low rates of discontinuation, and a high barrier to resistance.


Alternative strategies towards a sustainable municipal solid waste management system: A case study in Cairo.
Major cities in transitional and developing countries are facing the ever-growing challenge of managing solid waste in a sustainable manner. While a variety of treatments exist for solid waste, cities are in need of a sustainable integrated municipal solid waste management (MSWM) system. Such a system is meant to assist them in selecting and investing in an appropriate combination of treatments for the waste they generate, depending on the composition and quantity of this waste. This study presents a systems analysis of the MSWM in Cairo. A constrained non-linear mathematical model is developed to represent an underlying model of a MSWM system, with six waste material flows (cardboard and paper, plastics, metals, glass, organic material, and others). The developed model depicts combinations of five treatment alternatives (composting, anaerobic digestion, mechanical biological treatment, incineration, and landfilling). The treatment methods and their capacity are determined by the model, indicating possible optimal design solutions and recommendations. Starting with an evaluation of the status quo of the MSWM system in Cairo, the paper proposes a staged strategy, involving investigating improvements to the current mixed-waste management system, then exploring the possibility of introducing at-source waste sorting. The environmental and economic implications of different scenarios are analyzed and compared. A material flow analysis, including input data uncertainties, is also conducted by applying substance flow analysis. This analysis shows the difference between the status quo and the proposed improved solutions for diverting waste from landfills and, thus, decreasing carbon dioxide emissions.


PARROT: Prediction of enzyme abundances using protein-constrained metabolic models.
Protein allocation determines the activity of cellular pathways and affects growth across all organisms. Therefore, different experimental and machine learning approaches have been developed to quantify and predict protein abundance and how they are allocated to different cellular functions, respectively. Yet, despite advances in protein quantification, it remains challenging to predict condition-specific allocation of enzymes in metabolic networks. Here, using protein-constrained metabolic models, we propose a family of constrained-based approaches, termed PARROT, to predict how much of each enzyme is used based on the principle of minimizing the difference between a reference and an alternative growth condition. To this end, PARROT variants model the minimization of enzyme reallocation using four different (combinations of) distance functions. We demonstrate that the PARROT variant that minimizes the Manhattan distance between the enzyme allocation of a reference and an alternative condition outperforms existing approaches based on the parsimonious distribution of fluxes or enzymes for both Escherichia coli and Saccharomyces cerevisiae. Further, we show that the combined minimization of flux and enzyme allocation adjustment leads to inconsistent predictions. Together, our findings indicate that minimization of protein allocation rather than flux redistribution is a governing principle determining steady-state pathway activity for microorganism grown in alternative growth conditions.


Simian virus 5 V protein acts as an adaptor, linking DDB1 to STAT2, to facilitate the ubiquitination of STAT1.
The V protein of simian virus 5 (SV5) facilitates the ubiquitination and subsequent proteasome-mediated degradation of STAT1. Here we show, by visualizing direct protein-protein interactions and by using the yeast two-hybrid system, that while the SV5 V protein fails to bind to STAT1 directly, it binds directly and independently to both DDB1 and STAT2, two cellular proteins known to be essential for SV5-mediated degradation of STAT1. We also demonstrate that STAT1 and STAT2 interact independently of SV5 V and show that SV5 V protein acts as an adaptor molecule linking DDB1 to STAT2/STAT1 heterodimers, which in the presence of additional accessory cellular proteins, including Cullin 4a, can ubiquitinate STAT1. Additionally, we show that the avidity of STAT2 for V is relatively weak but is significantly enhanced by the presence of both STAT1 and DDB1, i.e., the complex of STAT1, STAT2, DDB1, and SV5 V is more stable than a complex of STAT2 and V. From these studies we propose a dynamic model in which SV5 V acts as a bridge, bringing together a DDB1/Cullin 4a-containing ubiquitin ligase complex and STAT1/STAT2 heterodimers, which leads to the degradation of STAT1. The loss of STAT1 results in a decrease in affinity of binding of STAT2 for V such that STAT2 either dissociates from V or is displaced from V by STAT1/STAT2 complexes, thereby ensuring the cycling of the DDB1 and SV5 V containing E3 complex for continued rounds of STAT1 ubiquitination and degradation.


The power of clinical nursing research: engage clinicians, improve patients' lives, and forge a professional legacy.
Sparked by the Institute of Medicine's report titled Crossing the Quality Chasm, research-based decision making has been emphasized for improving care. Patients should receive care that is based on the best available scientific knowledge, and care should not vary from clinician to clinician or from place to place. Implementing research-based practices at the bedside is a complex endeavor. It is all too easy to discover that clinically important research findings are either not known by practitioners or not being used in actual practice. Efforts to instill and sustain research-based practices improve significantly when staff nurses are involved with the research from the start. Institutions that are effective in involving clinicians have built a foundation of infrastructures that enable processes for engaging clinicians to take place. What distinguishes effective from ineffective hospital nursing research and evidence-based practice programs is the presence of structures whereby processes can occur that (1) unleash the creativity of staff by securing their involvement early, (2) educate staff by involving them, (3) create internal expertise for research and evidence-based practice, and (4) ensure that patients experience principled implementation of research-based practices to improve their lives. This article describes infrastructures that can ensure and sustain research-based practices while unleashing the talent and creativity of clinicians as they question practice and ponder the merits of current research. Fostering participation in such clinical inquiry will summon professional growth, influence the lives of patients, and help each nurse develop a unique personal professional legacy.


Expression of the neuron-specific potassium chloride cotransporter KCC2 in adult rat cochlear.
Auditory transduction in the cochlear is subject to modulate higher auditory centers in the brain via the efferent systems, which provide protection against damage caused by excessive excitation during auditory over stimulation. GABA is a proven inhibitory neurotransmitter in the efferent systems in mammalian cochlear. KCC2 is a neuron-specific potassium chloride cotransporter whose role in mature central neurons is to maintain the low intracellular Cl(-) concentrations required for the hyperpolarizing responses to the inhibitory amino acids GABA and glycine. However, there is a lack of information concerning KCC2 expression in the mammalian cochlear. In this study, reverse-transcription polymerase chain reaction (RT-PCR) and immunohistochemistry were used to detect the expression and localization of KCC2 in the mammalian cochlear. The results showed that these neuron-specific KCC2 transporters were present in most spiral ganglion neurons (SGNs) corresponding to the distribution of GABA(A)Rs. In addition, less intense reactions were observed on the organ of Corti, stria vascularis, and fibrocytes of the spiral ligament. These data suggest that KCC2 may play an important role in the modulation of a GABA neurotransmitter's function in a mammalian cochlear. Moreover, the presence of KCC2 on the organ of Corti and its surrounding tissues may contribute to maintaining normal K+ cycling. It is also presumed to be related to Cl(-) transportation in hair cells.


Using stereocartography for predicting efficacy of stereoinduction by chiral catalysts.
A computational method called stereocartography is used to examine regions around chiral catalysts that are most stereoinducing during Diels-Alder reactions. Geometries and atomic charges of catalysts are first generated quantum mechanically. The transition state of the reaction being catalyzed is then computed quantum mechanically and those enantiomeric transition states are used as probes to determine where around the catalyst stereoinduction is optimal. A description of how to treat catalysts with multiple conformations is given. In this article seven catalysts containing a variety of ligand motifs and metals were evaluated. The hypothesis that the region of maximum stereoinduction must be spatially coincident with the site of chemistry for a catalyst to be efficient is upheld.


Ultrasensitive amperometric determination of hand, foot and mouth disease based on gold nanoflower modified microelectrode.
Given the widespread use of point-of-care testing for diagnosis of disease, micro-scale electrochemical deoxyribonucleic acid (DNA) biosensors have become a promising area of research owing to its fast mass transfer, high current density and rapid response. In this study, a gold nanoparticles modified gold microelectrode (AuNPs/Au-Me) was constructed to determine the hand, foot and mouth disease (HFMD)-related gene. The noble metal nanoparticles modification yielded ca. 7.4-fold increase in electroactive surface area of microelectrode, and the signal for HFMD-related gene was largely magnified. Under optimal conditions, the biosensor exhibited salient selectivity and sensitivity with a low detection limit of 0.3 fM (S/N = 3), which is sufficient for clinical diagnosis of HFMD. Additionally, the developed AuNPs/Au-Me was successfully applied to determining the polymerase chain reaction (PCR) amplified products of target gene. Thus, the electrochemical DNA biosensor possesses great potential in early-stage diagnosis and long-term monitoring of various disease.


Use of a light-addressable potentiometric sensor for the detection of Escherichia coli O157:H7.
We describe the development of an immunoligand assay (ILA) in conjunction with a light-addressable potentiometric sensor (LAPS) for the rapid detection of Escherichia coli O157:H7 cells in buffered saline. The ILA protocol consists of "sandwiching" bacterial analyte between biotinylated and fluoresceinated antibodies, indirect enzyme labeling of the bacteria with urease-labeled anti-fluorescein antibody, and active capture of the immune complex at a biotinylated bovine serum albumin-blocked nitrocellulose filter membrane with streptavidin. Using live E. coli O157:H7, the efficiency of the ILA was compared using various ratios of the biotinylated and fluoresceinated antibodies. Simultaneous addition of equimolar biotinylated and fluoresceinated antibodies effected optimal urease labeling and subsequent active capture of the bacteria in the ILA. Equimolar concentrations of the antibodies were varied to achieve optimal LAPS detection response for the live bacteria. Using ILA with LAPS, a minimum detectable level of ca. 7.1 x 10(2) cells/ml of heat-killed or ca. 2.5 x 10(4) cells/ml of live E. coli O157:H7 bacteria was achieved in Tris-buffered saline in an assay time of ca. 45 or ca. 30 min, respectively.


Serious events with infliximab in patients with inflammatory bowel disease: a 9-year cohort study in the Netherlands.
The tumour necrosis factor-alpha inhibitor infliximab is incorporated in the treatment guidelines for patients with inflammatory bowel disease (IBD). However, concerns about serious adverse events such as infections, malignancies and death do exist. To evaluate the occurrence of serious events of infliximab during 9 years in a single-centre cohort of patients with IBD. Consecutive patients (>18 years) with a proven diagnosis of IBD who started treatment for IBD with infliximab at our referral centre in the Netherlands, from June 1999 to October 2007, were included. Infusion data were collected prospectively and medical records were reviewed retrospectively. All serious events were recorded and scored in the following categories: events leading to hospitalization, infections, malignancies and death. Severity and relationship to the use of infliximab were assessed for every serious event. 147 patients (33% male, mean age at first infusion 38 years, standard deviation = 12) received a total number of 1924 infusions (median per patient = 10, range 1-70). A total of 89 patients (61%) were hospitalized during follow-up, involving a total of 300 hospitalizations. Of these, 60 hospitalizations (20%) were considered at least possibly related to the use of infliximab. In 21 hospitalizations, the occurrence of a serious infection was considered at least possibly related to infliximab. Of all hospitalized patients, 70 patients (79%) underwent 139 surgical procedures, of which 70 surgeries (50%) were gastrointestinal related. Nine patients (6%) developed malignancies during follow-up: four colorectal carcinomas, one carcinoid tumour with another primary signet-ring cell carcinoma of the small bowel, one breast cancer, two skin cancers and one superficial melanoma. During follow-up, eight patients (5%) died: six as a result of malignancies, one patient as a result of a complication of short bowel syndrome and one patient due to unknown reasons. Patients who developed malignancies tended to have a longer disease duration than those who did not. Clinicians prescribing biological therapies should be aware of the development of serious events in their patients. Thorough follow-up of all patients during treatment with infliximab is warranted. If infliximab is considered in patients with IBD not responding to conventional treatment, efforts to exclude other possible underlying causes for worsening of symptoms should be made. Careful prescribing and monitoring during follow-up remains necessary.


Proton pump inhibitors enhance the effects of cytotoxic agents in chemoresistant epithelial ovarian carcinoma.
This study was designed to investigate whether proton pump inhibitors (PPI, V-ATPase blocker) could increase the effect of cytotoxic agents in chemoresistant epithelial ovarian cancer (EOC). Expression of V-ATPase protein was evaluated in patients with EOC using immunohistochemistry, and patient survival was compared based on expression of V-ATPase mRNA from a TCGA data set. In vitro, EOC cell lines were treated with chemotherapeutic agents with or without V-ATPase siRNA or PPI (omeprazole) pretreatment. Cell survival and apoptosis was assessed using MTT assay and ELISA, respectively. In vivo experiments were performed to confirm the synergistic effect with omeprazole and paclitaxel on tumor growth in orthotopic and patient-derived xenograft (PDX) mouse models. Expression of V-ATPase protein in ovarian cancer tissues was observed in 44 patients (44/59, 74.6%). Higher expression of V-ATPase mRNA was associated with poorer overall survival in TCGA data. Inhibition of V-ATPase by siRNA or omeprazole significantly increased cytotoxicity or apoptosis to paclitaxel in chemoresistant (HeyA8-MDR, SKOV3-TR) and clear cell carcinoma cells (ES-2, RMG-1), but not in chemosensitive cells (HeyA8, SKOV3ip1). Moreover, the combination of omeprazole and paclitaxel significantly decreased the total tumor weight compared with paclitaxel alone in a chemoresistant EOC animal model and a PDX model of clear cell carcinoma. However, this finding was not observed in chemosensitive EOC animal models. These results show that omeprazole pretreatment can increase the effect of chemotherapeutic agents in chemoresistant EOC and clear cell carcinoma via reduction of the acidic tumor microenvironment.


High-Throughput Mutant Screening via Transposon Sequencing.
Transposon mutagenesis has been the method of choice for genetic screens and selections in bacteria by virtue of the transposon being linked to the disrupted gene, simplifying its identification. Transposon sequencing (Tn-seq) is a high-throughput version of transposon mutant screening, in which massively parallel sequencing is used to simultaneously follow the fitness of all mutants in a complex library. In a single experiment, one can use Tn-seq to interrogate the contribution of all genes of a bacterium to fitness under a condition of interest. Here, we introduce a method to construct a saturating transposon insertion library in Gram-negative bacteria, to capture the transposon junctions en masse, and to identify essential genes and conditional genes using massively parallel sequencing. The accompanying protocol was developed as part of Cold Spring Harbor's Advanced Bacterial Genetics course.


Asymmetric Construction of 3,4-Diamino Pyrrolidines via Chiral N,O-Ligand/Cu(I) Catalyzed 1,3-Dipolar Cycloaddition of Azomethine Ylides with β-Phthalimidonitroethene.
A series of chiral N,O-ligands derived from a 1,2-dihydroimidazo[1,2-a]quinolone motif have been evaluated for the asymmetric 1,3-dipolar cycloaddition of azomethine ylides with a novel dipolarophile β-phthalimidonitroethene. A newly designed DHIPOH ligand 7c bearing 1-methyl and 4-iodo substituents was found to have significant "synergistic steric effects" and consequently afforded the corresponding 4-nitro-3-aminopyrrolidines with excellent diastereo- (dr up to 98:2) and enantio selectivities (ee up to 99%). Subsequent Raney Ni-catalyzed reduction and deprotection of phthalyl led to the structurally and biologically important 3,4-diaminopyrrolidines in a straightforward and efficient pathway.


Modeling the basin of attraction as a two-dimensional manifold from experimental data: applications to balance in humans.
We present a method of modeling the basin of attraction as a three-dimensional function describing a two-dimensional manifold on which the dynamics of the system evolves from experimental time series data. Our method is based on the density of the data set and uses numerical optimization and data modeling tools. We also show how to obtain analytic curves that describe both the contours and the boundary of the basin. Our method is applied to the problem of regaining balance after perturbation from quiet vertical stance using data of an elite athlete. Our method goes beyond the statistical description of the experimental data, providing a function that describes the shape of the basin of attraction. To test its robustness, our method has also been applied to two different data sets of a second subject and no significant differences were found between the contours of the calculated basin of attraction for the different data sets. The proposed method has many uses in a wide variety of areas, not just human balance for which there are many applications in medicine, rehabilitation, and sport.


Effects of previous dynamic arm exercise on power output during repeated maximal sprint cycling.
This study examined the effects of elevating blood lactate concentration by arm exercise on subsequent performance during repeated 30 s sprints with the legs. Eight male students performed two 30 s cycle ergometer sprints separated by 6 min of recovery, on two occasions. On one occasion the subjects performed only the two 30 s cycle ergometer sprints ('legs'), while on the other occasion 5 min of heavy arm cranking preceded the two sprints ('arms and legs'). Blood lactate concentration was determined from capillary samples at rest, after a standardized warm-up and 3 and 5 min following each exercise bout. In the 'legs' condition, the peak power output (PPO) and mean power output (MPO) in the second sprint were 92% (P < 0.05) and 85% (P < 0.01) of the values attained during the first sprint, respectively. Prior arm exercise, which increased blood lactate to 11.0 +/- 0.6 mM, had no effect on PPO and MPO during the first cycle ergometer sprint (approximately 4% drop, N.S.). However, in the second sprint after prior arm exercise, PPO was 10% lower than the PPO attained during the corresponding sprint in the 'legs' condition (sprint 2 'arms and legs' 963 +/- 42 W, sprint 2 'legs' 1074 +/- 60 W, P < 0.05), while MPO was better maintained (sprint 2 'arms and legs' 517 +/- 17 W, sprint 2 'legs' 549 +/- 24 W, N.S.). The rate of blood lactate accumulation after both cycle ergometer sprints was considerably decreased (by approximately 50%) when blood lactate levels were pre-elevated by arm crank exercise.(ABSTRACT TRUNCATED AT 250 WORDS)


Substance P and neurotensin in the control of gastrointestinal function.
Our experimental data show that neurotensin or a metabolite satisfies the rigid physiological criteria that should be met before a substance is classified as a true hormone with endocrine function. The gastrointestinal actions are produced by very small concentrations; the release into the blood is produced by a physiological stimulus (ingestion of food, especially fat); and the plasma concentrations of NTLI following the ingestion of food are comparable to those obtained by giving exogenous neurotensin at doses which produce effects in the gastrointestinal tract.


Meteorological conditions and incidence of Legionnaires' disease in Glasgow, Scotland: application of statistical modelling.
This study investigated the relationships between Legionnaires' disease (LD) incidence and weather in Glasgow, UK, by using advanced statistical methods. Using daily meteorological data and 78 LD cases with known exact date of onset, we fitted a series of Poisson log-linear regression models with explanatory variables for air temperature, relative humidity, wind speed and year, and sine-cosine terms for within-year seasonal variation. Our initial model showed an association between LD incidence and 2-day lagged humidity (positive, P = 0·0236) and wind speed (negative, P = 0·033). However, after adjusting for year-by-year and seasonal variation in cases there were no significant associations with weather. We also used normal linear models to assess the importance of short-term, unseasonable weather values. The most significant association was between LD incidence and air temperature residual lagged by 1 day prior to onset (P = 0·0014). The contextual role of unseasonably high air temperatures is worthy of further investigation. Our methods and results have further advanced understanding of the role which weather plays in risk of LD infection.


Effects of elevated temperatures on the embryonic development and corpus luteum activity in the Japanese long-fingered bat, Miniopterus schreibersii fuliginosus.
Gravid female Japanese long-fingered bats were kept in captivity without hibernation at elevated temperatures (23-25 degrees C) in winter. The embryonic growth rate was accelerated and consequently parturition was advanced by a period equivalent to that of exclusion from hibernation as compared with that in the wild population. The corpus luteum became active, as indicated by an increase in the 'light' lutein cells in an experimental bat pregnant with a 15-mm embryo, but was less active (more 'dark' cells) in 2 hibernating control bats with an implanting blastocyst.


Reversible Redox Activity in Multicomponent Metal-Organic Frameworks Constructed from Trinuclear Copper Pyrazolate Building Blocks.
Inorganic functionalization of metal-organic frameworks (MOFs), such as incorporation of multiple inorganic building blocks with distinct metals into one structure and further modulation of the metal charges, endows the porous materials with significant properties toward their applications in catalysis. In this work, by an exploration of the role of 4-pyrazolecarboxylic acid (H2PyC) in the formation of trinuclear copper pyrazolate as a metalloligand in situ, four new MOFs with multiple components in order were constructed through one-pot synthesis. This metalloligand strategy provides multicomponent MOFs with new topologies (tub for FDM-4 and tap for FDM-5) and is also compatible with a second organic linker for cooperative construction of complex MOFs (1,4-benzenedicarboxylic acid for FDM-6 and 2,6-naphthalenedicarboxylic acid for FDM-7). The component multiplicity of these MOFs originates from PyC's ability to separate Cu and Zn on the basis of their differentiated binding affinities toward pyrazolate and carboxylate. These MOFs feature reversible and facile redox transformations between CuI3(PyC)3 and CuII3(μ-OH)(PyC)3(OH)3 without altering the connecting geometries of the units, thus further contributing to the significant catalytic activities in the oxidation of CO and aromatic alcohols and the decomposition of H2O2. This study on programming multiple inorganic components into one framework and modulating their electronic structures is an example of functionalizing the inorganic units of MOFs with a high degree of control.


Microglia-Specific Expression of HEXA and HEXB Leads to Poor Prognosis in Glioblastoma Patients.
Glioblastoma multiforme (GBM) is one of the deadliest cancers in brain. There have been few treatment advances for GBM despite increasing scientific understanding of this disease. β-hexosaminidase (Hex) is an important enzyme system in human body, encoded by two genes, HEXA and HEXB, are closely related to central nervous system (CNS) diseases such as Sandhoff disease (SD) and Tay-Sachs disease (TSD). However, the expression pattern and function of HEXA and HEXB in GBM remains unclear. Here, we found that both the mRNA and protein expression levels of HEXA and HEXB were significantly upregulated in GBM patient samples. The results from single-cell RNA-sequencing (scRNA-seq) database and double immunostaining showed that HEXA and HEXB were specifically expressed in microglia in GBM patient samples. Furthermore, our in vitro experiments revealed that conditioned media from HEXA and HEXB knockdown-microglia cells could inhibit the proliferation and migration of GBM cells. Finally, according to survival analysis based on online database, higher expression of HEXA and HEXB was associated with poor prognosis in GBM patients. In conclusion, these results suggest that microglial HEXA and HEXB play fundamental role in GBM progression, and they will be potential biomarkers for GBM.


SARS Unique Domain (SUD) of Severe Acute Respiratory Syndrome Coronavirus Induces NLRP3 Inflammasome-Dependent CXCL10-Mediated Pulmonary Inflammation.
Severe acute respiratory syndrome-associated coronavirus (SARS-CoV) initiates the cytokine/chemokine storm-mediated lung injury. The SARS-CoV unique domain (SUD) with three macrodomains (N, M, and C), showing the G-quadruplex binding activity, was examined the possible role in SARS pathogenesis in this study. The chemokine profile analysis indicated that SARS-CoV SUD significantly up-regulated the expression of CXCL10, CCL5 and interleukin (IL)-1β in human lung epithelial cells and in the lung tissues of the mice intratracheally instilled with the recombinant plasmids. Among the SUD subdomains, SUD-MC substantially activated AP-1-mediated CXCL10 expression in vitro. In the wild type mice, SARS-CoV SUD-MC triggered the pulmonary infiltration of macrophages and monocytes, inducing CXCL10-mediated inflammatory responses and severe diffuse alveolar damage symptoms. Moreover, SUD-MC actuated NOD-, LRR- and pyrin domain-containing protein 3 (NLRP3) inflammasome-dependent pulmonary inflammation, as confirmed by the NLRP3 inflammasome inhibitor and the NLRP3-/- mouse model. This study demonstrated that SARS-CoV SUD modulated NLRP3 inflammasome-dependent CXCL10-mediated pulmonary inflammation, providing the potential therapeutic targets for developing the antiviral agents.


Climate-neutral and Smart Cities: a critical review through the lens of environmental justice.
The political choices made by the European institutions in the last twenty years show how the conviction is increasingly rooted that the management of environmental problems and, more specifically, the fight against climate change can find a valid solution in technology and eco-innovations. This is evident starting from the last two growth strategies adopted (Europe 2020 and the European Green Deal), from the long series of measures implemented to put them into practice and from the main R&I funding programs, such as Horizon Europe. In this context, the problem of justice and inclusiveness of the various initiatives implemented is attracting growing attention. In fact, if the institutional documents assume that green and smart participated projects are also fair and inclusive, a growing body of literature based on empirical studies seems to refute this assumption. Within this framework, the present work analyses first the critical literature and then the three main preparatory documents for the Horizon Europe Mission Climate-neutral and Smart Cities, which selected 100 European cities to become climate-neutral by 2030. These have been studied through the lens of environmental justice, in order to assess the European Commission's understanding of the existing and arising equity issues in the path toward climate neutrality. The research shows that, while the first two documents seemed informed by the idea that participation automatically translates into equality, the last guidelines show a deeper acknowledgement of the multidimensional nature of environmental justice. One that, beyond participation, also considers issues of distribution, rights, responsibilities and recognition. The present work should nevertheless be understood as a preparatory, analytical tool that will require the further definition and implementation of Climate City Contracts by the selected cities, in order to assess how the issue of environmental justice is effectively being considered in each specific context.


Influence of Intracanal Materials in Vertical Root Fracture Pathway Detection with Cone-beam Computed Tomography.
Investigating the vertical root fracture (VRF) pathway under different clinical scenarios may help to diagnose this condition properly. We aimed to determine the capability and intrareliability of VRF pathway detection through cone-beam computed tomographic (CBCT) imaging as well as analyze the influence of different intracanal and crown materials. VRFs were mechanically induced in 30 teeth, and 4 clinical situations were reproduced in vitro: no filling, gutta-percha, post, and metal crown. A Prexion (San Mateo, CA) 3-dimensional tomographic device was used to generate 104 CBCT scans. The VRF pathway was determined by using landmarks in the Avizo software (Version 8.1; FEI Visualization Sciences Group, Burlington, MA) by 1 observer repeated 3 times. Analysis of variance and post hoc tests were applied to compare groups. Intrareliability demonstrated an excellent agreement (intraclass correlation coefficient mean = 0.93). Descriptive analysis showed that the fracture line measurement was smaller in the post and metal crown groups than in the no-filling and gutta-percha groups. The 1-way analysis of variance test found statistically significant differences among the groups measurements. The Bonferroni correction showed statistically significant differences related to the no-filling and gutta-percha groups versus the post and metal crown groups. The VRF pathway can be accurately detected in a nonfilled tooth using limited field of view CBCT imaging. The presence of gutta-percha generated a low beam hardening artifact that did not hinder the VRF extent. The presence of an intracanal gold post made the fracture line appear smaller than it really was in the sagittal images; in the axial images, a VRF was only detected when the apical third was involved. The presence of a metal crown did not generate additional artifacts on the root surface compared to the intracanal gold post by itself.


[News in the treatment of hypertension and dyslipidemia].
These review article presents the results of major recent studies in cardiovascular prevention. HOPE-3 study showed that people with medium cardiovascular risk who have not experienced a cardiovascular event, and have normal blood pressure, benefit from deploying a statin, and in the presence of hypertension benefit from deploying a combination of a statin and antihypertensive drugs. Studies evaluating genetic predisposition brought new evidence that both LDL cholesterol and systolic blood pressure are independent and multiplicative causal risk factors for cardiovascular events. A mild long-term reduction in blood pressure and LDL cholesterol has the potential to significantly reduce the lifetime risk of cardiovascular events. In the final section the article discusses options for improving patient adherence to antihypertensive and lipid treatment and the new available fixed combinations for these diseases.Key words: arterial hypertension - cardiovascular prevention - dyslipidemia - fixed combinations - treatment.


Diet and drug interactions in the treatment of mental illness--a review.
Drug-diet interactions in the treatment of mental illness are reviewed from two perspectives: the potential effects of drugs on a person's nutritional status, and the effect of diet on drug action through modification of its absorption or various stages of its metabolism. As mental illness is often chronic and recurrent, requiring long-term drug therapy, it is important to become aware of the effects of drugs and their possible interactions with other important factors such as food.


Pembrolizumab related Guillain barre syndrome, a rare presentation in a patient with a history of lupus and bladder cancer.
Immune checkpoint inhibitor-related neurotoxicity causing Guillain Barre Syndrome is relatively uncommon. We discussed an 80-year-old patient with known systemic lupus erythematosus who presented with lower extremity weakness, areflexia and then progressed to respiratory muscle and upper extremity weakness after receiving immunotherapy with checkpoint inhibitors for metastatic bladder cancer. With the increasing use of immunotherapy for the management of cancer, awareness of neurological autoimmune side effects is essential. Immune checkpoint inhibitor-mediated GBS can be severe and fatal if not diagnosed promptly. The hospitalists, neurologists, and oncologists should be aware of neurotoxicity related to immune checkpoint inhibitor therapy requiring a multidisciplinary approach to patient care. Prompt initiation of immunosuppressive therapy is required for the management of immune checkpoint inhibitor-related neurotoxicity.


Evolution of rhizobia by acquisition of a 500-kb symbiosis island that integrates into a phe-tRNA gene.
Nodulation and nitrogen fixation genes of Mesorhizobium loti are encoded on the chromosome of the bacterium. Nevertheless, there is strong evidence that these genes can be transferred from an inoculant strain to nonsymbiotic mesorhizobia in the field environment. Here we report that the chromosomal symbiotic element of M. loti strain ICMP3153 is transmissible in laboratory matings to at least three genomic species of nonsymbiotic mesorhizobia. The element is 500 kb in size, integrates into a phe-tRNA gene, and encodes an integrase of the phage P4 family just within its left end. The entire phe-tRNA gene is reconstructed at the left end of the element upon integration, whereas the 3' 17 nucleotides of the tRNA gene are present as a direct repeat at the right end. We termed the element a symbiosis island on the basis of its many similarities to pathogenicity islands. It may represent a class of genetic element that contributes to microbial evolution by acquisition.


Characterization of the Maize Stalk Rot Pathogens Fusarium subglutinans and F. temperatum and the Effect of Fungicides on Their Mycelial Growth and Colony Formation.
Maize is a socioeconomically important crop in many countries. Recently, a high incidence of stalk rot disease has been reported in several maize fields in Gangwon province. In this report, we show that maize stalk rot is associated with the fungal pathogens Fusarium subglutinans and F. temperatum. Since no fungicides are available to control these pathogens on maize plants, we selected six fungicides (tebuconazole, difenoconazole, fluquinconazole, azoxystrobin, prochloraz and kresoxim-methyl) and examined their effectiveness against the two pathogens. The in vitro antifungal effects of the six fungicides on mycelial growth and colony formation were investigated. Based on the inhibition of mycelial growth, the most toxic fungicide was tebuconazole with 50% effective concentrations (EC50) of <0.1 μg/ml and EC90 values of 0.9 μg/ml for both pathogens, while the least toxic fungicide was azoxystrobin with EC50 values of 0.7 and 0.5 μg/ml for F. subglutinans and F. temperatum, respectively, and EC90 values of >3,000 μg/ml for both pathogens. Based on the inhibition of colony formation by the two pathogens, kresoxim-methyl was the most toxic fungicide with complete inhibition of colony formation at concentrations of 0.1 and 0.01 μg/ml for F. subglutinans and F. temperatum, respectively, whereas azoxystrobin was the least toxic fungicide with complete inhibition of colony formation at concentrations >3,000 μg/ml for both pathogens.


Dehydration and pancreatitis in a child lost in the mountains for 2 days.
An 8-year-old boy became lost while hiking in the Sierra Nevada Mountains of northern California in August 1994. He was found 2 days later, at which time he was dehydrated and admitted to the hospital for rehydration. After admission, he was found to also have mild pancreatitis. Although trauma could not be ruled out as the cause of the pancreatitis, the stress associated with being lost is felt to be an equally likely etiology.


Nonequilibrium stationary state of a harmonic crystal with alternating masses.
We analyze the nonequilibrium steady states (NESS) of a one-dimensional harmonic chain of N atoms with alternating masses connected to heat reservoirs at unequal temperatures. We find that the temperature profile defined through the local kinetic energy T(j)≡<pj2>/mj oscillates with period two in the bulk of the system. Depending on boundary conditions, either the heavier or the lighter particles in the bulk are hotter. We obtain explicit integral expressions for the bulk temperature profile and steady state current in the limit N→∞. These depend on whether N is odd or even. We also study similar temperature oscillations in the NESS of systems with noise in the dynamics. These die out as N→∞.


Estrogen effects on chromosome number and sister chromatid exchanges in uterine epithelial cells and kidney cells from neonatal mice.
Female mice of the NMRI strain were treated with a high dose (5 micrograms) or a low dose (10(-2) micrograms) of the synthetic estrogen diethylstilbestrol (DES) or estradiol-17 beta (E2) 48 and 24 hr, 24 hr, or 12 hr before death on day 3 after birth. Cultures of epithelium from the uterine cervix and uterine horns as well as of kidney cells were set up, and, after a culture period of 3 days, cells were studied for chromosome number and sister chromatid exchanges. Cervical cells from control females had a distinct peak of tetraploid cells, which was depressed after DES treatment but less so after E2 treatment. The tetraploid peak was lower in uterine horn epithelium and was less influenced by DES. No indications were obtained for an estrogen-induced aneuploid cell population in the uterine horn epithelium or cervical epithelium. The incidence of cells with a high number of sister chromatid exchanges (HFCECs) was only about one-fifth in uterine horn epithelium from control females compared with cervical epithelium from the same females. In the cervical epithelium, DES at both dose levels and E2 at the high dose induced an increased and similar incidence of HFCECs (from about 12.5% in controls to about 35%), which was not related to estrogen exposure time. In uterine horn epithelium, the incidence of HFCECs increased from 2.5% in controls to 8.5% after DES treatment. It is postulated that there is a more pronounced genomic plasticity in the cervical epithelium compared with the uterine horn epithelium. The estrogens had no effect on chromosome number or HFCECs in kidney cells.


Spatial information dynamics during early zebrafish development.
During development inherited information directs growth and specifies the complex spatial organization of cells and molecules. Here we show that a new information metric, the k-space information (kSI), captures the growth and emergence of spatial organization in a developing embryo. Using zebrafish as a model, we quantify the rate of development over the first 24h and demonstrate that important developmental landmarks are associated with well-defined transitions in information dynamics. The rate of development during this time is highest immediately before and after gastrulation, as well early in the segmentation period. We also find that the majority of the information arises from spatial correlations on the length scale of 20-80 μm, but there are contributions from many length scales that change over time. A comparison of the information dynamics in the maternal-zygotic one-eyed pinhead mutant, which is defective in mesoderm induction, with the wild-type embryo shows that the information dynamics diverge near the onset of gastrulation. Subsequently the mutant lacks a peak in the information dynamics that appears to be associated with the formation of trunk somites in the wild-type embryo. These findings provide a common and objective basis by which to quantify spatial organization, compare mutants and quantify developmental dynamics. The kSI can also be applied to any form of developmental data of arbitrary dimensions, and it offers a broad conceptual framework with which to organize the large amounts of data emerging from various sources.