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Background: Tuberculum sellae meningiomas (TSM) account for 3–10% of intracranial meningiomas. Visual loss is the presenting symptom in up to 80% of cases. Surgical management poses a great challenge due to tumor proximity to neurovascular structures such as the optic nerve and the internal carotid artery (ICA); hence, there is controversy regarding the optimal approach. The aim of this study is to determine differences in visual outcomes between transcranial (TCA) and endoscopic endonasal (EEA) approaches. Methods: A retrospective study including 29 patients with TSM surgically treated by TCA or EEA between 2011 and 2023 in a single referral center was conducted. Pre-and post-operative neuro-ophthalmologic evaluations, focusing on visual acuity and campimetry, were evaluated. Results: Sixteen (55.16%) patients were intervened through a TCA and the remaining 13 (44.84%) via an EEA. The lesions in each group were similar in terms of pre- operative volume (15.12 vs 12.9 cm3, p = 0.497) and neurovascular invasion (optic canal invasion 48.26 vs 41.37%, p = 0.664; ICA 44.81 vs 31.03%, p = 0.797). There were no significant differences in visual outcomes between both approaches; TCA presented an improvement of 5.18 points in visual fields (p = 0.140), whereas EEA had an improvement of 17.39 points in visual acuity (p = 0.114). Conclusion: EEA seems to offer greater improvement in visual acuity than TCA. However, the ideal approach should be individualized; taking into account the tumor’s volume and invasiveness, as well as the patient's visual complaints.

Mobile air pollution measurements have the potential to provide a wide range of insights into emission sources and air pollution exposure. The analysis of mobile data is, however, highly challenging. In this work we develop a new regression-based framework for the analysis of mobile data with the aim of improving the potential to draw inferences from such measurements. A quantile regression approach is adopted to provide new insight into the distribution of NOx and CO emissions in Central and Outer London. We quantify the emissions intensity of NOx and CO (ΔNOx/ΔCO2 and ΔCO/ΔCO2) at different quantile levels (τ) to demonstrate how transient high-emission events can be examined in parallel to the average emission characteristics. We observed a clear difference in the emissions behaviour between both locations. On average, the median (τ = 0.5) ΔNOx/ΔCO2 in Central London was 2x higher than Outer London, despite the stringent emission standards imposed throughout the Ultra Low Emissions Zone. A comprehensive vehicle emission remote sensing data set (n ≈ 700,000) is used to put the results into context, providing evidence of vehicle behaviour which is indicative of poorly controlled emissions, equivalent to high-emitting classes of older vehicles. Our analysis suggests the coupling of a diesel-dominated fleet with persistently congested conditions, under which the operation of emissions after-treatment technology is non-optimal, leads to increased NOx emissions.

The collective structure of 106Cd is elucidated by multi-step Coulomb excitation of a 3.849 MeV/A beam of 106Cd on a 1.1 mg/cm2 208Pb target using GRETINA-CHICO2 at ATLAS. Fourteen E2 matrix elements were obtained. The nucleus 106Cd is a prime example of emergent collectivity that possesses a simple structure: it is free of complexity caused by shape coexistence and has a small, but collectively active number of valence nucleons. This work follows in a long and currently active quest to answer the fundamental question of the origin of nuclear collectivity and deformation, notably in the cadmium isotopes. The results are discussed in terms of phenomenological models, the shell model, and Kumar-Cline sums of E2 matrix elements. The 〈02+||E2||21+〉 matrix element is determined for the first time, providing a total, converged measure of the electric quadrupole strength, 〈Q2〉, of the first-excited 21+ level relative to the 01+ ground state, which does not show an increase as expected of harmonic and anharmonic vibrations. Strong evidence for triaxial shapes in weakly collective nuclei is indicated; collective vibrations are excluded. This is contrary to the only other cadmium result of this kind in 114Cd by C. Fahlander et al. (1988) [38], which is complicated by low-lying shape coexistence near midshell.

Wilms tumor (WT) accounts for 90% of renal malignancies in children. Surgical resection is the mainstay of treatment, but neoadjuvant chemotherapy is considered in certain cases of tumor thrombus [1,2]. In North America, tumor biopsies are generally performed prior to initiating neoadjuvant chemotherapy. This report describes a case of a 7-year-old boy who presented with an abdominal mass and gross hematuria. An abdominal ultrasound and CT scan revealed a 9 cm right-sided renal mass most consistent with Wilms tumor. A near obstructing tumor thrombus was identified in the inferior vena cava (IVC) from the right renal vein to the confluence of the hepatic veins. In consultation with oncology and according to Children's Oncology Group (COG) recommendations [2], a biopsy was performed prior to initiating neoadjuvant chemotherapy. An endovascular, transvenous biopsy of the tumor thrombus was performed. In this case, a transvenous biopsy of the IVC tumor thrombus was an alternative technique that yielded adequate tissue to establish the diagnosis of Wilms tumor including histologic sub-classification and LOH status.

A 17-year-old male with Osteogenesis Imperfecta (OI) type 1 and history of prior spinal instrumentation for kyphosis presented with severe symptomatic pectus excavatum (PEx). His Haller index was 6.7 with a Pectus Correction Index of 51%. He underwent uncomplicated Nuss procedure with insertion of 3 bars and complete repair of the PEx. A single sternal fracture was sustained during correction and healed without deformity. After two years, he developed left-sided pain and two of 3 bars were removed with maintained chest correction. This case report demonstrates the feasibility of Nuss procedure in adolescents with OI and outlines improvements to the technique in future patients with OI.

Proton inelastic scattering off a neutron halo nucleus, 11Li, has been studied in inverse kinematics at the IRIS facility at TRIUMF. The aim was to establish a soft dipole resonance and to obtain its dipole strength. Using a high quality 66 MeV 11Li beam, a strongly populated excited state in 11Li was observed at Ex=0.80±0.02 MeV with a width of Γ=1.15±0.06 MeV. A DWBA (distorted-wave Born approximation) analysis of the measured differential cross section with isoscalar macroscopic form factors leads us to conclude that this observed state is excited in an electric dipole (E1) transition. Under the assumption of isoscalar E1 transitions, the strength is evaluated to be extremely large amounting to 30∼296 Weisskopf units, exhausting 2.2%∼21% of the isoscalar E1 energy-weighted sum rule (EWSR) value. The large observed strength originates from the halo and is consistent with the simple di-neutron model of 11Li halo.

A multi-step Coulomb excitation measurement with the GRETINA and CHICO2 detector arrays was carried out with a 430-MeV beam of the neutron-rich 110Ru (t1/2=12 s) isotope produced at the CARIBU facility. This represents the first successful measurement following the post-acceleration of an unstable isotope of a refractory element. The reduced transition probabilities obtained for levels near the ground state provide strong evidence for a triaxial shape; a conclusion confirmed by comparisons with the results of beyond-mean-field and triaxial rotor model calculations.

The ellipsoid body (EB) of the insect brain performs pivotal functions in controlling navigation. Input and output of the EB is provided by multiple classes of R-neurons (now referred to as ER-neurons) and columnar neurons which interact with each other in a stereotypical and spatially highly ordered manner. The developmental mechanisms that control the connectivity and topography of EB neurons are largely unknown. One indispensable prerequisite to unravel these mechanisms is to document in detail the sequence of events that shape EB neurons during their development. In this study, we analyzed the development of the Drosophila EB. In addition to globally following the ER-neuron and columnar neuron (sub)classes in the spatial context of their changing environment we performed a single cell analysis using the multi-color flip out (MCFO) system to analyze the developmental trajectory of ER-neurons at different pupal stages, young adults (4d) and aged adults (∼60d). We show that the EB develops as a merger of two distinct elements, a posterior and anterior EB primordium (prEBp and prEBa, respectively. ER-neurons belonging to different subclasses form growth cones and filopodia that associate with the prEBp and prEBa in a pattern that, from early pupal stages onward, foreshadows their mature structure. Filopodia of all ER-subclasses are initially much longer than the dendritic and terminal axonal branches they give rise to, and are pruned back during late pupal stages. Interestingly, extraneous branches, particularly significant in the dendritic domain, are a hallmark of ER-neuron structure in aged brains. Aging is also associated with a decline in synaptic connectivity from columnar neurons, as well as upregulation of presynaptic protein (Brp) in ER-neurons. Our findings advance the EB (and ER-neurons) as a favorable system to visualize and quantify the development and age-related decline of a complex neuronal circuitry., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Objectives: We propose fast and accurate molecular detection of the Y132F ERG11p substitution directly on pure-cultured Candida parapsilosis isolates. We also assessed a discriminative genotyping scheme to track circulating genotypes., Methods: A total of 223 C. parapsilosis isolates (one patient each) from 20 hospitals, located in Spain and Italy were selected. Isolates were fluconazole-resistant (n = 94; harbouring the Y132F ERG11p substitution [n = 85], the G458S substitution [n = 6], the R398I substitution [n = 2], or the wild-type ERG11 gene sequence) or fluconazole-susceptible (n = 129). Two targeted-A395T-mutation PCR formats (conventional and real-time) were engineered and optimized on fluconazole-susceptible and fluconazole-resistant pure-cultured isolates, thus skipping DNA extraction. Two genotyping schemes were compared: Scheme 1 (CP1, CP4a, CP6, and B markers), and Scheme 2 (6A, 6B, 6C, CP1, CP4a, and CP6 markers)., Results: The screening performed using both PCR formats showed 100% specificity (fluconazole-susceptible isolates; n = 129/129) and sensitivity (Y132F isolates; n = 85/85) values; however, results were available in 3 and 1.5 hours with the conventional and real-time PCR formats, respectively. Overall, Scheme 1 showed higher genetic diversity than Scheme 2, as shown by the number of alleles detected (n = 98; mean 23, range 13-38), the significantly higher observed and expected heterozygosity, and the probability of identity index (2.5 × 10 -6 ). Scheme 2 markers did not provide further genotypic discrimination of Y132F fluconazole-resistant genotypes., Conclusion: Both proposed PCR formats allow us to speed up the accurate detection of substitution Y132F ERG11p in C. parapsilosis isolates with 100% specificity and sensitivity. In addition, we recommend CP1, CP4a, CP6, and B microsatellite markers for genotyping fluconazole-resistant isolates., (Copyright © 2024 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

Objectives: To analyze the hypertensive disorders of pregnancy (HDP) trends in the 2000-2020 period and its association with social determinants of health (SDH) in Mexican municipalities., Study Design: This was a retrospective longitudinal study analyzing data from routine information systems on 21,493,076 hospital discharges due to maternal causes and SDH data of 2,439-2,450 municipalities between 2000 and 2020., Main Outcomes Measures: We used joinpoint regression models to analyze HDP trends and hybrid negative binomial regression models to examine the association between SDH and HDP rates at intra- and inter-municipality levels., Results: HDP rate increased from 0.54 in 2000 to 2.42 cases per 100 live births in 2020 (annual average percent change of 7.7 %, 95 % CI 6.6-8.9). Municipalities with higher marginalization index had higher HDP rates (IRR 1.12, 95 % CI 1.01-1.24; 1.36, 95 % CI 1.25-1.47; 1.30, 95 % CI 1.23-1.37, and 1.06, 95 % CI 1.00-1.12, in 2000-2004, 2005-2009, 2010-2014, 2015-2020, respectively). Municipal increases in the percentage of population with high schooling were associated with decreases in HDP rates in 2000-2009. Meanwhile, the role of socioeconomic status at the state level was less consistent, yet the HDP rates were lower in most socially advantaged regions., Conclusion: During 21 years, HDP rates increased overall in Mexican municipalities, suggesting an increased maternal morbidity burden due to HDP. The association of municipal marginalization and socioeconomic level with HDP trends indicates a need to ensure equitable interventions to improve maternal health in Mexican municipalities., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

In this work nanocomposites based on alginate (Alg) and halloysite as a nanotubular clay (Hy) were developed. Characterization techniques reveal that Hy/Alg nanocomposites are cation exchangers with predominantly negative charge density and good thermal stability. The adsorption equilibrium of Cd(II) in aqueous solution onto Hy/Alg nanocomposites revealed that by increasing the mass of halloysite in the nanocomposite, the adsorption capacity diminished significantly due to the halloysite-alginate interactions. Maximum adsorption capacities of 8, 65, 88, and 132 mg/g of Cd(II) were obtained for samples Hy, Hy/Alg 50%, Hy/Alg 95%, and Alg, respectively. In addition, the adsorption equilibrium of Cd(II) on the Hy/Alg bionanocomposites was affected by the pH and temperature of the solution, demonstrating the presence of electrostatic interactions during adsorption and that this is an exothermic process. The controlling mechanism of adsorption was cation exchange influenced by electrostatic forces. The Cd(II) adsorption rate studies were interpreted by the diffusion-permeation model and reveal that the presence of Hy in the structure of the nanocomposites enhances the permeation coefficient, that is, the adsorption rate was increased. The values of the permeation coefficient varied from 1.95 × 10 -7 to 8.50 × 10 -7  cm 2 /s for Hy/Alg 50% and from 1.70 × 10 -7 to 3.55 × 10 -7  cm 2 /s for Hy/Alg 95%., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Studies are urgently needed to characterize immunogenicity, efficacy, and safety of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) mRNA vaccines in kidney transplant (KT) recipients, excluded from major clinical trials. Complex ELISPOT and other cellular response techniques have been applied, but simpler tools are needed. An easy-to-use real-world monitoring of SARS-CoV-2 IgG antibodies against the Spike protein and QuantiFERON ® SARS-CoV-2 IFNγ release assay (IGRA) were performed at baseline and 28 days after the second dose in KT recipients and controls (dialysis patients and healthy ones). All healthy controls and >95% dialysis controls became positive for anti-S IgG antibodies, while only 63.3% of KT patients seroconverted with a very low antibody level. A positive IGRA was documented in 96.9% of controls, 89.3% peritoneal dialysis, 77.6% hemodialysis, 61.3% of KT patients transplanted more than 1 year ago and only 36% of those transplanted within the previous 12 months. Overall, 100% of healthy controls, 95.4% of dialysis patients and 78.8% KT recipients developed any immune response (humoral and/or cellular) against SARS-CoV-2. KT patients showed low rates of immune responses to mRNA Coronavirus infectious disease 2019 vaccines, especially those with recent transplantations. Simple humoral and cellular monitoring is advisable, so that repeated doses may be scheduled according to the results., (© 2021 The American Society of Transplantation and the American Society of Transplant Surgeons.)

Objectives: The aim of this study was to evaluate the accuracy of various susceptibility methods when testing cefuroxime against a collection of Escherichia coli isolates with MIC values close to the breakpoint., Methods: 80 E. coli strains with a cefuroxime MIC value of 16 mg/L obtained by broth microdilution with Vitek 2 were selected. Microdilution was considered the reference standard and was performed in duplicate, as were disc and gradient diffusion tests using two different manufacturers in each case. EUCAST 8.0 breakpoints were used for MIC interpretation., Results: All strains were resistant according to Vitek 2 (MIC 16 mg/L) but 72.5% (58/80) were classified as susceptible by reference standard microdilution. Categorical and essential agreements between Vitek 2 and reference standard microdilution were 27.5% (95% CI 1.9-1.4) and 86.3% (95% CI 0.8-0.9), respectively. Differences are statistically significant when isolates are classified as 'susceptible' or 'resistant' according to EUCAST breakpoints between diffusion methods (disc and gradient) and reference standard microdilution. Using BioMérieux (BM) and Liofilchem (LF) gradient testing, 24.1% (14/58) and 13.8% (8/58) of results were identified as false susceptible and 4.5% (1/22) and 40.9% (9/22) were found to be false resistant, respectively. Using Oxoid (OX) and Bio Rad (BR) cefuroxime discs, 22.5% (13/58) and 17.2% (10/58) of results were false susceptible and 9.1% (2/22) and 13.6% (3/22) were false resistant, respectively., Discussion: Intertechnique variation around the cefuroxime breakpoint was a considerable source of disagreements and seriously affected the clinical classification of the isolates. We propose that the definition of the area of technical uncertainty (ATU) be modified to include the variability between approved AST methods., (Copyright © 2020 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

Dientamoeba fragilis is a trichomonad parasite of the human intestine that is found worldwide. However, the biological cycle and transmission of this parasite have yet to be elucidated. Although its pathogenic capacity has been questioned, there is increasing evidence that clinical manifestations vary greatly. Different therapeutic options with antiparasitic drugs are currently available; however, very few studies have compared the effectiveness of these drugs. In the present longitudinal study, we evaluate 13,983 copro-parasitological studies using light microscopy of stools, during 2013-2015, in Terrassa, Barcelona (Spain). A total of 1150 (8.2%) presented D. fragilis. Of these, 739 episodes were finally analyzed: those that involved a follow-up parasitology test up to 3 months later, corresponding to 586 patients with gastrointestinal symptoms (53% under 15 years of age). Coinfection by Blastocystis hominis was present in 33.6% of the subjects. Our aim was to compare therapeutic responses to different antiparasitic drugs and the factors associated with the persistence of D. fragilis post-treatment. Gender, age, and other intestinal parasitic coinfections were not associated with parasite persistence following treatment. Metronidazole was the therapeutic option in most cases, followed by paromomycin: 65.4% and 17.5% respectively. Paromomycin was found to be more effective at eradicating parasitic infection than metronidazole (81.8% vs. 65.4%; p = 0.007), except in children under six years of age (p = 0.538). Although Dientamoeba fragilis mainly produces mild clinical manifestations, the high burden of infection means we require better understanding of its epidemiological cycle and pathogenicity, as well as adequate therapeutic guidelines in order to adapt medical care and policies to respond to this health problem., (Copyright © 2019 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Objectives: To determine if telepharmacy can be used to collect medication histories on patients admitted in the emergency department (ED) in a large health system., Practice Description: As part of an effort to address safety concerns, resource limitations, and a decline in medication history completions, a program was developed to use telepharmacy to conduct medication histories on patients admitted in the ED., Setting: The medication history program covers 5 large facilities. It is staffed by 6 full-time pharmacy technicians 7 days a week and is overseen by a pharmacist., Innovation: Medication histories are conducted with the use of mobile carts enhanced with videoconferencing equipment allowing technicians to operate from a remote central location. The program allows the technicians to observe multiple EDs at one time, interview patients through videoconferencing, and document completed medication histories in the electronic medical record (EMR). The technicians also transcribe preoperation (pre-op) medication lists for patients being admitted for surgeries., Evaluation: Medication history completion rates and barriers were assessed. In addition, potential medication errors, medication history accuracy rates for nursing and pharmacy technicians, and a cost analysis of preventable ADEs were explored., Results: The program, on average, conducts medication histories on 56% to 79% of patients admitted through the ED during hours of operation. In fiscal year 2018, the technicians entered 24,980 medication histories and pre-op lists. A cross-sectional analysis of data collected from December 2016 to March 2017, including 124 patients, revealed 320 potential medication errors among a total of 382 high-risk medications., Conclusion: Based on the current performance and continued expansion of this novel strategy, use of telepharmacy to obtain medication histories in the ED has led to resource optimization for the remote delivery of a pharmacy service., (Copyright © 2019 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.)

In this work chili seeds (Capsicum annuum) were used as raw material in the synthesis of biochar at temperatures between 400 and 600 °C. The samples were chemically, texturally and morphologically characterized and their properties were correlated with the calcination temperature. The adsorption mechanism of IBP was elucidated by analyzing the effect of solution pH, ionic strength and temperature, whereas that, the intraparticle diffusion mechanism was clarified through the application of a 3D diffusional model. The results evidenced that raising the pyrolysis temperature promotes a greater content of disordered graphitic carbon (51.6-85.02%) with small surface area (0.52-0.18 m 2 /g) and low quantity of functional groups. The adsorption study demonstrated that the biochar synthesized at 600 °C (C600) enhances the adsorption capacity >50 folds compared with chili seeds. Moreover, at pH = 7 the adsorption mechanism is governed by π-acceptor and attractive electrostatic interactions, whereas at basic pH the main adsorption mechanism is π-acceptor. Additionally, hydrophobic interactions become important by increasing the presence of NaCl. The application of 3D diffusional model based on surface diffusion interpreted clearly the kinetic curves obtaining values of D s ranging from 2.31 × 10 -8 -2.51 × 10 -8  cm 2  s -1 . Besides, it was determined that intraparticle mass flux is larger along the shortest axis of the seed, and always directed toward the particle center. The maximum mass flux takes place in the center of particle, and it advances like a moving front as time was increased., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The competitive sorption of the antibiotics sulfamethoxazole (SMX) and trimethoprim (TMP) and SMX-Cd(II) on a hybrid clay nanosorbent (NanoSorb) was investigated in detail in this work. NanoSorb was synthesized by sorbing a surfactant on bentonite. Besides, the sorption of SMX on the NanoSorb was confirmed by FTIR analysis, and SMX was mainly sorbed on NanoSorb by a partition mechanism due to hydrophobic interactions. Otherwise, the single adsorption of Cd(II) and TMP onto NanoSorb were due to electrostatic interaction and hydrophobic partition, respectively. The capacity of NanoSorb for sorbing single SMX was very similar to that for single Cd(II), but more than 10 times higher than that for single TMP. The competitive sorption of SMX-TMP was antagonistic because the sorption of one antibiotic on NanoSorb was decreased by the presence of the other antibiotic. The uptake of SMX was reduced up to 43.4% by the presence of TMP, whereas the presence of SMX decreased the uptake of TMP up to 29.6%. The non-modified Langmuir multicomponent isotherm (NLMI) interpreted quite well the experimental competitive sorption data of SMX-TMP. On the other hand, the competitive sorption of SMX-Cd(II) on NanoSorb revealed that the sorption of SMX was non-interactive because it was not influenced by the presence of Cd(II). Whereas, the sorption of Cd(II) was synergistic or cooperative since the uptake of Cd(II) sorbed increased considerably with the uptake of SMX sorbed on NanoSorb. The two-site Langmuir model fitted the experimental competitive sorption data of Cd(II) on NanoSorb saturated with SMX. The application of this isotherm was based on the fact that Cd(II) sorbed on two types of sites: a) cationic sites of the NanoSorb and b) Pi-cation interactions between the aromatic ring of the SMX sorbed on NanoSorb and Cd 2+ ., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Objectives: The aim of this study was to monitor the spread, persistence and antibiotic resistance patterns of Legionella spp. strains found in a hospital water distribution system. These environmental studies are intended to help detect the presence of antibiotic resistant strains before they infect patients., Methods: Antimicrobial surveillance tests were performed at 27 different sampling points of the water network of a large Spanish hospital over two years. Water samples were screened for Legionella according to ISO 11731:2007. Legionella spp. isolates were identified by serotyping and by mass spectrometry (MALDI-ToF). Epidemiological molecular typing was done by Pulse-Field Gel Electrophoresis (PFGE) and by Sequence-Based Typing (SBT). Antibiotic susceptibility tests were performed using disk diffusion and ETEST®., Results: Legionella spp. were recurrently isolated for 2 years. All isolates belonged the same group, L. pneumophila serogroups 2-14. Isolates were all attributed by SBT to sequence type (ST) ST328, although PFGE revealed 5 different patterns. No significant change in antibiotic susceptibility could be observed for this study period, irrespectively of the method used., Conclusion: Colonization of water systems by Legionella spp. is still occurring, although all the prevention rules were strictly followed. Antibiotic resistance monitoring may help us to find resistance in bacteria with environmental reservoirs but difficult to isolate from patients. The knowledge of the antibiotic susceptibility in environmental strains may help us to predict changes in clinical strains. This study might also help reconsidering Legionnaires' disease (LD) diagnostic methods. L. pneumophila serogroups 2-14 present all along the time of the investigation in the water distribution system can cause LD. However, they may not be detected by routine urine tests run on patients, thereby missing an ongoing LD infection., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Repetitive oscillations in cytoplasmic Ca 2+ due to periodic Ca 2+ release from the endoplasmic reticulum (ER) drive mammalian embryo development following fertilization. Influx of extracellular Ca 2+ to support the refilling of ER stores is required for sustained Ca 2+ oscillations, but the mechanisms underlying this Ca 2+ influx are controversial. Although store-operated Ca 2+ entry (SOCE) is an appealing candidate mechanism, several groups have arrived at contradictory conclusions regarding the importance of SOCE in oocytes and eggs. To definitively address this question, Ca 2+ influx was assessed in oocytes and eggs lacking the major components of SOCE, the ER Ca 2+ sensor STIM proteins, and the plasma membrane Ca 2+ channel ORAI1. We generated oocyte-specific conditional knockout (cKO) mice for Stim1 and Stim2, and also generated Stim1/2 double cKO mice. Females lacking one or both STIM proteins were fertile and their ovulated eggs displayed normal patterns of Ca 2+ oscillations following fertilization. In addition, no impairment was observed in ER Ca 2+ stores or Ca 2+ influx following store depletion. Similar studies were performed on eggs from mice globally lacking ORAI1; no abnormalities were observed. Furthermore, spontaneous Ca 2+ influx was normal in oocytes from Stim1/2 cKO and ORAI1-null mice. Finally, we tested if TRPM7-like channels could support spontaneous Ca 2+ influx, and found that it was largely prevented by NS8593, a TRPM7-specific inhibitor. Fertilization-induced Ca 2+ oscillations were also impaired by NS8593. Combined, these data robustly show that SOCE is not required to support appropriate Ca 2+ signaling in mouse oocytes and eggs, and that TRPM7-like channels may contribute to Ca 2+ influx that was previously attributed to SOCE., (Published by Elsevier Ltd.)

Fifty-seven nosocomial Klebsiella pneumoniae isolates producing extended-spectrum beta-lactamases (ESBLs) were collected between February 2007 and November 2007 in different wards of the Sarajevo (Bosnia-Herzegovina) reference hospital. These isolates comprise two major epidemic pulsed-field electrophoresis-defined clones plus two minor clones. In addition to the ESBL-mediated resistance, all strains uniformly showed resistance to ciprofloxacin, gentamicin and tobramycin. The beta-lactamases involved in this resistance phenotype were TEM-1, SHV-1, and CTX-M-15, as demonstrated by isoelectric focusing, PCR amplification, and sequencing. TEM-1 and CTX-M-15 beta-lactamases, as well as the aminoglycoside resistance determinants, were encoded in plasmids that could be transferred to Escherichia coli by conjugation. In three of the infected patients with the predominant clone, cefoxitin resistance development (MICs >128 mg/L) was documented. The analysis of the outer membrane proteins of the cefoxitin-susceptible and cefoxitin-resistant isolates revealed that the former expressed only one of the two major porins, OmpK36, whereas in the latter, the expression of Ompk36 was altered or abolished. This is the first report of CTX-M-15-producing K. pneumoniae in Bosnia-Herzegovina. Furthermore, we document and characterize for the first time cefoxitin resistance development in CTX-M-15-producing K. pneumoniae.

Clinical isolates (n = 389) of methicillin-resistant Staphylococcus aureus (MRSA) recovered from 371 patients between January 2003 and June 2004 at the three major public hospitals on the island of Majorca, Spain were studied. The clonal relatedness of MRSA isolates was determined by pulsed-field gel electrophoresis (PFGE) after digestion with SmaI. During the study period, MRSA was found in 31% of patients with S. aureus-positive cultures. PFGE analysis identified three predominant clones, affecting 94% of the patients. The three clones had been detected since 1999 in one hospital, and were designated as clones A, B and C. Whereas clones A and B (multidrug-resistant) were related to the two most prevalent clones in Spain at this time, clone C was identical to EMRSA-15, currently one of the most common MRSA clones in UK hospitals and also detected in other countries, but rarely in Spanish hospitals. This imported epidemic clone was detected in c. 10% of patients admitted to one of the three hospitals in 2002, but its prevalence has increased significantly (32% of the patients investigated in the three hospitals in the present study), and this clone also accounted for 44% of the isolates from non-hospitalised patients. Even though EMRSA-15 showed the least multidrug resistance of the three major clones, it was apparently more virulent, since it was associated significantly (p 0.001) with bacteraemia, and positive blood cultures were documented for 21% of the patients infected by this clone, compared with only 10% and 7% of patients infected with clones A and B, respectively.

This study investigated the efficacy of a commercial enzyme immunoassay (Directigen RSV, ColorPAC) in comparison with the shell vial culture method (using Hep-2 cells) for the detection of respiratory syncytial virus (RSV) in nasopharyngeal aspirates from children with bronchiolitis. During the period 1995-2002, 4950 samples were examined. RSV was detected in 1660 (33.5%) samples, with a sensitivity of 80.9%, a specificity of 97.5%, a positive predictive value of 93.8%, a negative predictive value of 91.6%, and a testing efficiency value of 92.2% compared with shell vial culture. In 83 (5%) samples, the ColorPAC was positive and the shell vial assay was negative. Of these, 71 (85.6%) were false-negative by cell culture. The true false-positive results obtained by ColorPAC represented only 0.7% of all RSV-positive samples. In general, no statistically significant differences were detected between the different months and epidemic periods studied. Compared with ColorPAC, the shell vial culture method displayed a sensitivity of 95.8% and a specificity of 100%. Overall, the ColorPAC assay was an acceptable, simple and rapid method for the antigenic detection of RSV in paediatric respiratory samples.

The use of natural antioxidants and flavonoids in nutritional and pharmaceutical applications is increasing. Because some phytochemicals such as genistein, found in soy products, have estrogenic activity, we investigated the estrogenic potential of a natural antioxidant mixture (NAO) isolated from spinach leaves, using an in vivo uterotrophic bioassay and an in vitro transcriptional activation assay for the estrogen receptor (ER). Outbred female CD-1 mice (17 d old) were given subcutaneous injections of 17beta-estradiol or genistein [500 and 500,000 microg /(kg x d), respectively] as positive controls or NAO [1000 to 1,000,000 microg/(kg x d)] for 3 d. Uterine wet weight/body weight ratios were determined. Both 17beta-estradiol and genistein significantly increased uterine wet weight ratios compared with untreated controls, but NAO did not. Histological examination of the uterus showed that 17beta-estradiol and genistein increased epithelial cell height, number and gland development, but NAO did not. Estrogenic activity of NAO was investigated in vitro using the ER transcriptional activation assay. BG1Luc4E2 cells expressing ER were stably transfected with a luciferase reporter gene responsive to estrogens. 17beta-estradiol dose dependently increased luciferase activity; NAO had no effect. When NAO was tested for antiestrogenic activity, it did not lessen the effects of 17beta-estradiol. These data suggest that NAO does not have estrogenic or antiestrogenic activity. Thus, an antioxidant mixture has been identified that does not have potentially adverse estrogenic activity.

The estrogenic responses of several phytoestrogens including genistein, daidzein, coumestrol, alpha-zearalanol, zearalenone, naringenin, taxifolin and biochanin A were compared over a wide dose range using an in vitro assay that measures transcriptional activation of the estrogen receptor (ER) and an in vivo immature mouse uterotrophic assay consisting of measuring uterine wet weight increase plus sensitive morphological and biochemical endpoints in the uterus. The transcriptional activation assay showed activation of the ER by all compounds tested except taxifolin with varying magnitudes of response as compared to estradiol or diethylstilbestrol. Results from the uterotropic bioassay showed that genistein, coumestrol, zearalanol, and zearalenone caused an increase in uterine wet weight, while naringenin, taxifolin, daidzein and biochanin A failed to do so over the dose range tested. However, sensitive morphological and biochemical parameters such as uterine epithelial cell height increase, uterine gland number increase, and induction of the estrogen-responsive protein lactoferrin demonstrated that all compounds tested in this study gave some measure of estrogenicity although a wide range of estrogenic responses across compounds was shown. Use of multiple in vitro and in vivo estrogenic endpoints as described in this paper will be useful in developing estrogenic profiles for individual compounds and ultimately mixtures of compounds. Furthermore, having an estrogenic "fingerprint" for each phytochemical is an essential first step in determining potential adverse effects of exposure to phytoestrogens.

The aim of this study was to compare the clinical, biological, and radiologic features of presentation in the emergency ward of community-acquired pneumonia (CAP) by Legionella pneumophila (LP) and other community-acquired bacterial pneumonias to help in early diagnosis of CAP by LP. Three hundred ninety-two patients with CAP were studied prospectively in the emergency department of a 600-bed university hospital. Univariate and multivariate analyses were performed to compare epidemiologic and demographic data and clinical, analytical, and radiologic features of presentation in 48 patients with CAP by LP and 125 patients with CAP by other bacterial etiology (68 by Streptococcus pneumoniae, 41 by Chlamydia pneumoniae, 5 by Mycoplasma pneumoniae, 4 by Coxiella burnetii, 3 by Pseudomonas aeruginosa, 2 by Haemophilus influenzae, and 2 by Nocardia species. Univariate analysis showed that CAP by LP was more frequent in middle-aged, male healthy (but alcohol drinking) patients than CAP by other etiology. Moreover, the lack of response to previous beta-lactamic drugs, headache, diarrhea, severe hyponatremia, and elevation in serum creatine kinase (CK) levels on presentation were more frequent in CAP by LP, while cough, expectoration, and thoracic pain were more frequent in CAP by other bacterial etiology. However, multivariate analysis only confirmed these differences with respect to lack of underlying disease, diarrhea, and elevation in the CK level. We conclude that detailed analysis of features of presentation of CAP allows suspicion of Legionnaire's disease in the emergency department. The initiation of antibiotic treatment, including a macrolide, and the performance of rapid diagnostic techniques are mandatory in these cases.

Reactive oxygen species (ROS) have been implicated in the pathophysiology of renal ischemia/reperfusion injury. Endothelin-1 (ET-1) is generated in abundance in renal ischemia/reperfusion with resultant decreases in renal blood flow and glomerular filtration rate. To determine if ROS regulate ET-1 production, the effect of ROS donors or scavengers on ET-1 protein and mRNA levels in cultured human mesangial cells was examined. Incubation with xanthine/xanthine oxidase, glucose oxidase, or H2O2 caused a dose-dependent rise in ET-1 release. Similarly, xanthine/xanthine oxidase or H2O2 augmented ET-1 mRNA levels. In contrast, the ROS scavengers dimethylthiourea (DMTU), dimethylpyrroline N-oxide, or pyrrolidine dithiocarbamate reduced basal ET-1 release, while DMTU lowered ET-1 mRNA levels. Deferoxamine, an iron chelator, also decreased basal ET-1 release. Superoxide dismutase potentiated the ET-1 stimulatory effect of xanthine/xanthine oxidase, while catalase abrogated the effect of xanthine/xanthine oxidase and H2O2. The effects of ROS were unrelated to changes in nitric oxide production or cytotoxicity. These data indicate that exogenously or endogenously-derived ROS can increase ET-1 production by human mesangial cells. While superoxide anion reduces ET-1 levels, H2O2 leads to enhanced production of the peptide. ROS stimulation of mesangial cell ET-1 production may contribute to impaired glomerular hemodynamics in the setting of renal ischemia/reperfusion injury.

Prostaglandin E2 (PGE2) may be an important negative feedback modulator of endothelin-1 (ET-1)-stimulated mesangial cell proliferation and contraction. Recent studies suggest that ET-1 may induce prolonged mesangial cell PGE2 production, however the mechanism of this effect is unknown. The current study was undertaken, therefore, to examine the long-term effect of ET-1 on mesangial cell PGE2 synthesis. ET-1 markedly increased PGE2 release by rat mesangial cells for at least six hours. Cyclooxygenase (COX) activity was increased by one hour and persisted for at least six hours. ET-1 increased COX-2, but not COX-1, protein and mRNA levels. Actinomycin D reduced ET-1-stimulated PGE2 synthesis and COX-2 mRNA expression, while cycloheximide superinduced COX-2 mRNA. Dexamethasone decreased ET-1-stimulated PGE2 release and COX-2 protein and mRNA levels. ET-1-stimulated PGE2 release was prevented by BQ-123, an endothelin receptor A antagonist. We conclude that ET-1, via activation of the endothelin A receptor, causes a prolonged increase in mesangial cell PGE2 production that is partially dependent on induction of dexamethasone-inhibitable COX-2.