Sustained virologic response to treatment in 100% of patients recently infected, nosocomially, with HCV genotype 2.
In 2003, a cluster of hepatitis C virus (HCV)-infected patients with a common history of a surgical procedure, performed during 2001 to 2003, was identified in a medical center. An epidemiologic investigation linked a physician, infected with HCV genotype 2, as the possible source of infection in 35 patients. The evaluation, therapy, and outcome of this unique cohort are presented. HCV-RNA was isolated from sera of all patients and the double-stranded phosphorylation homology domain region was sequenced. After a routine clinical investigation 33 patients were offered antiviral therapy. Two patients were not treatment candidates due to old age and comorbidity. Twenty-two (66%) were women. The mean age was 48.5+/-16.9 years. Alanine aminotransferase level was 117+/-135 IU/L. Thirty patients were treated with pegylated interferon alpha 2a, 1 with pegylated interferon alpha 2b, and 1 with standard interferon. All received ribavirin 800 mg daily. One patient refused to be treated and was lost for follow-up. Time from acquisition of disease to initiation of therapy was 14.8+/-4.9 month (5.5 to 26). Therapy duration was 24 weeks except for 1 patient who stopped therapy after 16 weeks. Sustained virologic response was achieved in all 32 treated patients. The sequence motif of the phosphorylation homology domain region, studied in all patients, predicted good response to interferon. Our excellent results can be explained by a constellation of favorable viral characteristics, a short-term disease and adherence to therapy.


High-throughput ligand profile characterization in novel cell lines expressing seven heterologous insect olfactory receptors for the detection of volatile plant biomarkers.
Agriculturally important crop plants emit a multitude of volatile organic compounds (VOCs), which are excellent indicators of their health status and their interactions with pathogens and pests. In this study, we have developed a novel cellular olfactory panel for detecting fungal pathogen-related VOCs we had identified in the field, as well as during controlled inoculations of several crop plants. The olfactory panel consists of seven stable HEK293 cell lines each expressing a functional Drosophila olfactory receptor as a biosensing element along with GCaMP6, a fluorescent calcium indicator protein. An automated 384-well microplate reader was used to characterize the olfactory receptor cell lines for their sensitivity to reference VOCs. Subsequently, we profiled a set of 66 VOCs on all cell lines, covering a concentration range from 1 to 100 μM. Results showed that 49 VOCs (74.2%) elicited a response in at least one olfactory receptor cell line. Some VOCs activated the cell lines even at nanomolar (ppb) concentrations. The interaction profiles obtained here will support the development of biosensors for agricultural applications. Additionally, the olfactory receptor proteins can be purified from these cell lines with sufficient yields for further processing, such as structure determination or integration with sensor devices.


Safety of Ventricular Reservoir Sampling in Pediatric Posthemorrhagic Hydrocephalus Patients: Institutional Experience and Review of the Literature.
INTRODUCTION: Posthemorrhagic hydrocephalus (PHH) is a common disease process encountered in neonates. Management often includes cerebrospinal fluid (CSF) aspiration through ventricular access devices (VADs). However, a common concern surrounding serial access of implanted subcutaneous reservoirs includes introduction of infection. In addition, there is great variability in aseptic technique. Therefore, the authors sought to evaluate the incidence of VAD access-associated infections in the literature and compare them with the rate of infection found at our institution. We also highlight the use of a preassembled VAD access kit and standardized access protocol, as well as the role of provider education, in maintaining safety and sterility during serial VAD access. METHODS: A single-institution retrospective review was performed for PHH patients younger than 1 year old undergoing serial CSF aspirations via implanted VADs (2009-2019). Patients were excluded if they had a ventriculoperitoneal shunt placed as primary intervention. MEDLINE search for reports of serial VAD access in PHH was also performed. Reports were excluded if they did not include full-text articles in the English literature. RESULTS: At our institution, subcutaneous reservoirs were placed in 37 neonates with PHH for serial CSF aspiration. No infections occurred after a total of 630 taps (average, 17 taps per reservoir; range, 0-83) and 10 420 collective reservoir days (average, 282 per patient; range, 6-3700). Only 2 reservoirs required revision for malfunction. Serial VAD taps for PHH were described in 14 articles in the medical literature, with 7.9% (n = 47/592) of patients reported with tap-related infectious complications. CONCLUSION: A standardized VAD access kit, along with stringent adherence to access protocol, can significantly minimize risk of infection associated with serial VAD access. These principles can be generalized to percutaneous aspiration of CSF from subcutaneous reservoirs placed for other indications to promote safety and sterility of this common procedure.


Secukinumab treatment of moderate to severe plaque psoriasis in routine clinical care: real-life data of prior and concomitant use of psoriasis treatments from the PROSPECT study.
Secukinumab, a fully human anti-interleukin-17A monoclonal antibody, has demonstrated efficacy and safety in patients with moderate to severe psoriasis. However, as per study protocols, transition periods from prior psoriasis treatments of a defined minimal length were required and use of concomitant psoriasis medication was prohibited. There is therefore a lack of data on the effect of shorter transition periods and concomitant psoriasis treatment with other pharmacologically active substances on the effectiveness and safety of secukinumab in routine clinical practice. The PROSPECT study was designed to assess prior and concomitant use of psoriasis treatments in subjects receiving secukinumab and the duration of transition periods from prior treatments to secukinumab. Here, we report the baseline characteristics and the duration of transition period in an interim analysis of the first 805 subjects. PROSPECT is an ongoing 24-week, single-cohort, non-interventional study. Subjects with moderate to severe psoriasis with a decision to receive secukinumab were included. The majority of subjects were male (491/796, 61.7%), with a mean age of 47.7 years (SD 13.7). The baseline Psoriasis Area and Severity Index (PASI) was available for 92.4% (744/805) of subjects, and mean baseline PASI was 17.5 (SD 13.1); 93.4% (752/805) of subjects had signs of high disease severity. Use of concomitant treatment increased with the number of signs. Within the last 12 months prior to inclusion, 10%, 40%, and 28% of subjects had received topical, conventional systemic, or biologic treatments as their last prior psoriasis therapy, respectively, and 22% of subjects had not received any psoriasis therapy. Discontinuation of prior treatment due to adverse events was high in subjects with conventional systemic treatment (93/413, 22.5%) compared to biologic treatment (5/210, 2.4%). The median duration of the transition period was 14.0, 30.5, and 38.0 days for prior topical, conventional systemic, and biologic treatments, respectively. PROSPECT is the first study to investigate prior and concomitant use of psoriasis treatments in subjects receiving secukinumab in a real-world setting. The majority of the subjects had a high disease burden and use of concomitant treatment increased with disease severity. The duration of the transition period depended on prior treatment.


Medication overuse headache in 787 patients admitted for inpatient treatment over a period of 32 years.
Definitions of medication overuse headache have changed over time. To evaluate the clinical characteristics of medication overuse headache patients admitted for inpatient withdrawal therapy over a period of 32 years. We included all patients with medication overuse headache treated from 1 January 1984 to 31 December 2015. We obtained all data from the medical reports and defined three periods, P1 (1984-1993), P2 (1994-2003), and P3 (2004-2015). The p-value adjusted for multiple comparisons was set to 0.005. Within 32 years, a total of 787 patients accounted for 904 admissions for MOH. From P1 to P3, the proportion of patients with preexisting migraine increased from 44.3% to 53.3% (chi2 = 9.0, p = 0.01) and that with preexisting tension-type headache decreased from 47.9% to 34.6% (chi2 = 9.3, p < 0.01). The median time since onset of headache and medication overuse headache decreased from 20 to 15 years (p < 0.001) and from 3 to 2 years (p < 0.001). The median cumulative number of single doses decreased from 120 to 90 per month (p = 0.002). Overuse of triptans, non-opioid analgesics, and opioids increased, whereas overuse of ergotamines decreased over time (p < 0.001 for all tests). The use of prophylactic medication before admission increased from 8.3% to 29.9% (chi2 = 89.5, p < 0.001). This retrospective study in a large number of patients with medication overuse headache admitted for inpatient withdrawal therapy over a period of 32 years shows a trend towards changes in the preexisting headache type, a decrease in the time since onset of headache and medication overuse headache, a decrease in the number of drug doses used per month, changes in the type of drugs overused, and an increase in, but still low rate, of prophylactic medication prior to admission.


Could inducible protein-10 and heparin-binding hemagglutinin improve the detection of Mycobacterium tuberculosis-infected subjects in a country with low incidence of tuberculosis?
This study aimed to evaluate inducible protein-10 (IP-10) as a biomarker besides interferon-gamma (IFN-γ) to improve the identification of active tuberculosis (TB) and latent tubercular infection (LTBI) in a country with a low incidence of TB. Whole blood from Mycobacterium tuberculosis-infected subjects was stimulated with region-of-difference-1 (RD1)-specific peptides and with heparin-binding hemagglutinin (HBHA) to determine the release of IP-10 and IFN-γ. No statistically significant difference was observed between positive rates of IP-10 and IFN-γ after RD1-specific peptide stimulation in the TB and LTBI groups; a different response was detected in QuantiFERON TB-gold test-negative (QFT-) subjects. A significantly different proportion of positive responses was observed between IP-10 and IFN-γ following HBHA stimulation in the TB group and in the QFT- group but not in the LTBI group. The IP-10 test seemed to identify false-negative QFT results in some subjects with a positive IFN-γ/IP-10/HBHA pattern.


The efficacy of procalcitonin as a biomarker in the management of sepsis: slaying dragons or tilting at windmills?
Sepsis is defined as systemic inflammatory response syndrome (SIRS) in the context of an underlying infectious process, and is associated with high rates of morbidity and mortality, particularly when initial therapy is delayed. Numerous biomarkers, including but not limited to cytokines (interleukins-2 and -6 [IL-2, IL-6] and tumor necrosis factor-α [TNF-α]), leukotrienes, acute-phase proteins (C-reactive protein [CRP]), and adhesion molecules, have been evaluated and rejected as unsuitable for the diagnosis of sepsis, predicting its severity, and guiding its treatment. Most recently, procalcitonin (PCT) has been suggested as a novel biomarker that may be useful in guiding therapeutic decision making in the management of sepsis. This article assesses critically the published literature on the clinical utility of PCT concentrations for guiding the treatment of sepsis in adult patients. A comprehensive search of all published studies of the use of serum concentrations of PCT to guide the treatment of sepsis in adult patients (1996 to 2011) was conducted with PubMed and Google Scholar. The search focused on the value of PCT concentrations to guide the diagnosis, prognosis, monitoring, and escalation and de-escalation of antbiotic therapy in these patients. Keywords searched included "procalcitonin," "sepsis," "sepsis biomarker," "sepsis diagnosis," "sepsis prognosis," "sepsis mortality," "antibiotic escalation," "antibiotic de-escalation," "antibiotic duration," and "antimicrobial stewardship." Forty-six trials evaluating the efficacy of PCT concentrations in diagnosing sepsis have been published, with 39 of these trials yielding positive results and 7 yielding negative results. Wanner et al. published the largest study (n=405) demonstrating that peak PCT concentrations occur early after injury in both patients with sepsis and those with multiple organ dysfunction syndrome (MODS). Among 17 trials assessing the prognostic value of PCT concentrations with regard to clinical outcome and morbidity, 12 trials yielded positive results and five showed negative or equivocal results. Reith et al. published the largest study of the prognostic use of PCT concentrations (n=246), demonstrating that median PCT values on post-operative days (POD) one, four, and 10 were predictive of mortality in patients with abdominal sepsis (p<0.01). Among 14 trials of the utility of PCT concentrations for establishing an infectious cause of sepsis, 13 yielded positive results and only one yielded negative results. The largest study of this use of PCT concentrations, conducted by Baykut et al. (n=400), evaluated these concentrations in post-operative patients with infection, and demonstrated that concentrations of PCT remained elevated until POD 4, with a second increase observed between POD 4 and POD 6. In uninfected patients, PCT concentrations began to decrease on POD 2. Only a single study has assessed the utility of PCT concentrations in guiding the escalation of antibiotic therapy, and its results were negative. Specifically, Jensen et al. (n=1,200) compared a PCT-guided antibiotic escalation strategy with the standard of care for sepsis and found no difference in outcomes. They also found that the PCT group had a longer average stay in the intensive care unit (ICU), greater rates of mechanical ventilation, and a decreased estimated glomerular filtration rate (eGFR). Among four trials focusing on PCT concentrations and antibiotic de-escalation, all showed positive results with the measurement of PCT concentrations. The largest such study, by Bouadma et al. (n=621), demonstrated a four-day decrease in antibiotic duration when PCT concentrations were used to guide therapy relative to the study arm given the standard of care, with no increase in mortality (p=0.003). The diagnostic value of serum PCT concentrations for discriminating among SIRS, sepsis, severe sepsis, and septic shock remains to be established. Although higher PCT concentrations suggest a systemic bacterial infection as opposed to a viral, fungal, or inflammatory etiology of sepsis, serum PCT concentrations do not correlate with the severity of sepsis or with mortality. At present, PCT concentrations are solely investigational with regard to determining the timing and appropriateness of escalation of antimicrobial therapy in sepsis. Nevertheless, serum PCT concentrations have established utility in monitoring the clinical response to medical and surgical therapy for sepsis, and in surveillance for the development of sepsis in burn and ICU patients, and may have a role in guiding the de-escalation of antibiotic therapy.


Ventral acetabular augmentation for management of caudoventral luxation following total hip replacement in dogs.
To report ventral acetabular augmentation with an internal fixator for management of caudoventral luxation following total hip replacement in dogs and to report clinical outcomes. Clinical records from three UK-based referral hospitals were reviewed retrospectively between 2010 and 2020 to identify dogs with caudoventral hip luxation managed by ventral acetabular augmentation. Hip prosthesis component orientation was radiographically assessed to identify potential risk factors associated with the luxation. Clinical and radiographic assessments were performed at short-term (≤12 weeks) and long-term (>12 months) follow-up and all complications were recorded. A standardised owner telephone questionnaire was used to assess long-term outcomes when a clinical assessment was unavailable. Nine dogs were included. Risk factors contributing to the luxation could not be defined. Implants used included a string-of-pearls plate (6/9), a polyaxial locking plate combined with a polyethylene implant (1/9) or a polyethylene implant alone (2/9). Complications occurred in two of nine dogs, including recurrent luxation and femoral stem loosening; both dogs had received a polyethylene implant alone, of which one received explantation of the hip prosthesis. Median long-term follow-up was 39 months (range 13.5-62). Seven dogs who received a ventral acetabular plate achieved full function of the operated limb and the overall outcome was good for eight dogs. Follow-up radiographs revealed stable hip prostheses in dogs who received a ventral plate, with the longest assessment at 37 months postoperatively. Owner satisfaction was good for all cases. Ventral acetabular augmentation can successfully manage caudoventral luxation following total hip replacement in dogs.


Novel bispecific antibodies increase γδ T-cell cytotoxicity against pancreatic cancer cells.
The ability of human γδ T cells from healthy donors to kill pancreatic ductal adenocarcinoma (PDAC) in vitro and in vivo in immunocompromised mice requires the addition of γδ T-cell-stimulating antigens. In this study, we demonstrate that γδ T cells isolated from patients with PDAC tumor infiltrates lyse pancreatic tumor cells after selective stimulation with phosphorylated antigens. We determined the absolute numbers of γδ T-cell subsets in patient whole blood and applied a real-time cell analyzer to measure their cytotoxic effector function over prolonged time periods. Because phosphorylated antigens did not optimally enhance γδ T-cell cytotoxicity, we designed bispecific antibodies that bind CD3 or Vγ9 on γδ T cells and Her2/neu (ERBB2) expressed by pancreatic tumor cells. Both antibodies enhanced γδ T-cell cytotoxicity with the Her2/Vγ9 antibody also selectively enhancing release of granzyme B and perforin. Supporting these observations, adoptive transfer of γδ T cells with the Her2/Vγ9 antibody reduced growth of pancreatic tumors grafted into SCID-Beige immunocompromised mice. Taken together, our results show how bispecific antibodies that selectively recruit γδ T cells to tumor antigens expressed by cancer cells illustrate the tractable use of endogenous γδ T cells for immunotherapy.


Mechanisms of N-acetyl-p-benzoquinone imine cytotoxicity.
N-Acetyl-p-benzoquinone imine (NAPQI), a reactive metabolite of acetaminophen, rapidly reacts at physiological pH with glutathione (GSH) forming an acetaminophen-glutathione conjugate and stoichiometric amounts of acetaminophen and glutathione disulfide (GSSG). The same reaction products are formed in isolated hepatocytes incubated with NAPQI. In hepatocytes which have been treated with 1,3-bis-(2-chloroethyl)-1-nitrosourea (BCNU) in order to inhibit glutathione reductase, the initial rise in GSSG concentration in the presence of NAPQI is maintained, whereas GSSG is rapidly reduced back to GSH in untreated hepatocytes. Oxidation by NAPQI of GSH to GSSG and the reduction of GSSG back to GSH by the NADPH-dependent glutathione reductase appear to be responsible for the rapid oxidation of NADPH that occurs in hepatocytes incubated with NAPQI in that the effect is blocked by pretreatment of cells with BCNU. When added to hepatocytes, NAPQI not only reacts with GSH but also causes a loss in protein thiol groups. The loss in protein thiols occurs more rapidly in cells pretreated with BCNU or diethylmaleate. Whereas both of these treatments enhance cytotoxicity caused by NAPQI, BCNU pretreatment has no effect on the covalent binding of [14C-ring]NAPQI to cellular proteins. Furthermore, dithiothreitol added to isolated hepatocytes after maximal covalent binding of [14C-ring]NAPQI but preceding cell death protects cells from cytotoxicity and regenerates protein thiols. Thus, the toxicity of NAPQI to isolated hepatocytes may result primarily from its oxidative effects on cellular proteins.


MIRIAM: A machine and deep learning single-cell segmentation and quantification pipeline for multi-dimensional tissue images.
Increasingly, highly multiplexed tissue imaging methods are used to profile protein expression at the single-cell level. However, a critical limitation is the lack of robust cell segmentation tools for tissue sections. We present Multiplexed Image Resegmentation of Internal Aberrant Membranes (MIRIAM) that combines (a) a pipeline for cell segmentation and quantification that incorporates machine learning-based pixel classification to define cellular compartments, (b) a novel method for extending incomplete cell membranes, and (c) a deep learning-based cell shape descriptor. Using human colonic adenomas as an example, we show that MIRIAM is superior to widely utilized segmentation methods and provides a pipeline that is broadly applicable to different imaging platforms and tissue types.


Iron release from recombinant N-lobe and single point Asp63 mutants of human transferrin by EDTA.
Transferrins bind ferric ion and deliver the iron to cells. The mechanism of the iron release has been studied kinetically, in vitro, with the aid of single point mutants in which the iron-binding ligand, Asp63 (aspartic acid-63, D63), has been changed to Ser, Asn, Glu and Ala. Iron release from the unmutated N-lobe of human serum transferrin (hTF/2N) by EDTA is influenced by a variety of factors. The rate-determining conformational-change mechanism may be a major pathway for iron release from hTF/2N's having a 'closed' conformation, which leads to a saturation kinetic mode with respect to ligand concentration. The effect of chloride depends on the protein conformation, showing a negative action in the case of tight binding and a positive action when the protein has an 'open' or 'loose' conformation. The negative effect of chloride could originate from the binding competition between chloride and the chelate to the active site for iron release, and the positive effect could derive from the synergistic participation of chloride in iron removal. The 'open' conformation may be induced by decreasing pH: the transitional point appears to be at about pH 6.3 for the wild-type hTF/2N; the 'loose' conformation may be facilitated by mutations at D63, which result in the loss of a key linking component in interdomain interactions of the protein. In the latter case, structural factors dominate over other potential negative effects because the weak interdomain contacts derived from the mutation of D63 cause the binding site to open easily, even at pH 7.4. Therefore chloride exhibits an accelerating action on iron release by EDTA from all the D63 mutants.


Two distinct Ca(2+) signaling pathways modulate sperm flagellar beating patterns in mice.
Hyperactivation, a swimming pattern of mammalian sperm in the oviduct, is essential for fertilization. It is characterized by asymmetrical flagellar beating and an increase of cytoplasmic Ca(2+). We observed that some mouse sperm swimming in the oviduct produce high-amplitude pro-hook bends (bends in the direction of the hook on the head), whereas other sperm produce high-amplitude anti-hook bends. Switching direction of the major bends could serve to redirect sperm toward oocytes. We hypothesized that different Ca(2+) signaling pathways produce high-amplitude pro-hook and anti-hook bends. In vitro, sperm that hyperactivated during capacitation (because of activation of CATSPER plasma membrane Ca(2+) channels) developed high-amplitude pro-hook bends. The CATSPER activators procaine and 4-aminopyridine (4-AP) also induced high-amplitude pro-hook bends. Thimerosal, which triggers a Ca(2+) release from internal stores, induced high-amplitude anti-hook bends. Activation of CATSPER channels is facilitated by a pH rise, so both Ca(2+) and pH responses to treatments with 4-AP and thimerosal were monitored. Thimerosal triggered a Ca(2+) increase that initiated at the base of the flagellum, whereas 4-AP initiated a rise in the proximal principal piece. Only 4-AP triggered a flagellar pH rise. Proteins were extracted from sperm for examination of phosphorylation patterns induced by Ca(2+) signaling. Procaine and 4-AP induced phosphorylation of proteins on threonine and serine, whereas thimerosal primarily induced dephosphorylation of proteins. Tyrosine phosphorylation was unaffected. We concluded that hyperactivation, which is associated with capacitation, can be modulated by release of Ca(2+) from intracellular stores to reverse the direction of the dominant flagellar bend and, thus, redirect sperm.


Impact of the COVID-19 Pandemic on Surgical Volumes Among Fellowship-Trained Glaucoma Subspecialists.
The change in glaucoma surgical volumes due to the COVID-19 pandemic was not uniform across procedure types and unequal between rural and urban practice locations. To quantify the impact of the COVID-19 pandemic on surgical volumes performed by fellowship-trained glaucoma subspecialists. This retrospective cohort analysis of the CMS Medicare Public Use File extracted all glaucoma surgeries including micro-invasive glaucoma surgeries, trabeculectomy, goniotomy, lasers, and cataract surgery performed by fellowship-trained glaucoma surgeons in rural and urban areas between 2016 and 2020. Predicted estimates of 2020 surgical volumes were created utilizing linear squares regression. Percent change between predicted and observed 2020 surgical volume estimates was analyzed. Statistical significance was achieved at P<0.05. In 2020, fellowship trained glaucoma surgeons operated mostly in urban areas (N=810, 95%). A 29% and 31% decrease in predicted cataract surgery volumes in urban and rural areas was observed. Glaucoma surgeries experienced a 36% decrease from predicted estimates (N=56,781). MIGS experienced a 86% and 75% decrease in rural and urban areas respectively. Trabeculectomy in rural areas experienced a 16% increase relative to predicted estimates while urban areas experienced a decrease of 3% (P>0.05). The number of goniotomies decreased by 10% more in rural areas than in urban areas (-22% and -12%, respectively). Laser procedures decreased by 8% more in urban areas than in rural areas (-18% and -10%, respectively). Among glaucoma-trained surgeons, glaucoma surgeries experienced a greater volume loss than cataract surgeries. In urban US areas, relative reductions in MIGS and goniotomy volumes in urban areas may have been compensated by greater laser and trabeculectomy volumes. Trabeculectomies in rural areas was the only group exceeding predicted estimates. Glaucoma subspecialists may utilize these findings when planning for future events and in overcoming any remaining unmet need in terms of glaucoma care.


Smoking habits among medical students in Western Saudi Arabia.
To estimate the prevalence of tobacco smoking, and understand the attitude, practice, and knowledge among medical students. A cross-sectional study was conducted from September 2009 to May 2010. An anonymous, self-administered, Global Adult Tobacco Survey based questionnaire was completed by the students attending the main Medical College of King Abdulaziz University, Jeddah, Kingdom of Saudi Arabia. Of the 643 students in the study, 90 students (14%) indicated that they smoked tobacco at the time of the study. The prevalence of smoking was 24.8% among males, and 9.1% among females. Smoking was more common in males (p=0.003), but there were more ex-smokers among females (p=0.042). The friends and parents were considered the primary influence for initiating smoking habit, followed by the media. Ninety percent thought that doctors should set a good example by not smoking. Most of the study population indicated that smoking is related to serious illnesses; however, non-smokers were better aware of such illnesses than smokers. Although most thought that smoking tobacco is harmful, approximately 9.5% believe that smoking a water pipe is not. Despite the good knowledge on the hazards of tobacco consumption, 24.8% male, and 9.1% female medical students in Jeddah continue to smoke. The policymakers should address the factors contributing to the initiation, continuation, and spread of this devastating habit.


Severe acute respiratory syndrome-associated coronavirus 3a protein forms an ion channel and modulates virus release.
Fourteen ORFs have been identified in the severe acute respiratory syndrome-associated coronavirus (SARS-CoV) genome. ORF 3a of SARS-CoV codes for a recently identified transmembrane protein, but its function remains unknown. In this study we confirmed the 3a protein expression and investigated its localization at the surface of SARS-CoV-infected or 3a-cDNA-transfected cells. Our experiments showed that recombinant 3a protein can form a homotetramer complex through interprotein disulfide bridges in 3a-cDNA-transfected cells, providing a clue to ion channel function. The putative ion channel activity of this protein was assessed in 3a-complement RNA-injected Xenopus oocytes by two-electrode voltage clamp. The results suggest that 3a protein forms a potassium sensitive channel, which can be efficiently inhibited by barium. After FRhK-4 cells were transfected with an siRNA, which is known to suppress 3a expression, followed by infection with SARS-CoV, the released virus was significantly decreased, whereas the replication of the virus in the infected cells was not changed. Our observation suggests that SARS-CoV ORF 3a functions as an ion channel that may promote virus release. This finding will help to explain the highly pathogenic nature of SARS-CoV and to develop new strategies for treatment of SARS infection.


Macroscopic relations for microscopic properties at the interface between solid substrates and dense fluids.
Strongly confined fluids exhibit inhomogeneous properties due to atomistic structuring in close proximity to a solid surface. State variables and transport coefficients at a solid-fluid interface vary locally and become dependent on the properties of the confining walls. However, the precise mechanisms for these effects are not known as of yet. Here, we make use of nonequilibrium molecular dynamics simulations to scrutinize the local fluid properties at the solid-fluid interface for a range of surface conditions and temperatures. We also derive microscopic relations connecting fluid viscosity and density profiles for dense fluids. Moreover, we propose empirical ready-to-use relations to express the average density and viscosity in the channel as a function of temperature, wall interaction strength, and bulk density or viscosity. Such relations are key to technological applications such as micro-/nanofluidics and tribology but also natural phenomena.


Electron-microscopical approach to a structural model of intima collagen.
Intima collagen was studied by electron microscopy (rotary shadowing and negative staining) and by analytical ultracentrifugation. It was found that the monomeric unit (Mr 170 000) consists of a 105 nm-long triple helix terminated by a small globular domain (Mr about 30 000) at one end and a large globular domain (Mr about 40 000) at the other end. The monomer was produced by selective reduction of interchain disulphide bridges. Before reduction, dimers, tetramers and larger filamentous structures were found. Dimers are lateral staggered aggregates of two monomers aligned in an anti-parallel fashion. This gives rise to an inner 75 nm-long region of two slightly intertwisted triple helices flanked by the large globular domains. The outer triple-helical segments (length 30 nm) with the small globular domains at their ends emerge at both sides of this structure. Interchain disulphide bridges are probably located in the vicinity of the large domains. Only the outer segments could be degraded by bacterial collagenase. In tetramers the outer segments of two dimers are covalently linked, forming a scissors-like structure. In the fibrous forms several tetramers are assembled end-to-end with an overlap between the outer segments. The molecular masses and sedimentation coefficients were calculated for these various forms from the electron-microscopically observed dimensions and agreed with results obtained by ultracentrifugation. The unique structure of intima collagen suggests that it originates from a microfibrillar component and that it can be considered a unique collagenous protein, for which we propose the designation type VI collagen.


The etiology of social change.
A fundamental aspect of human beings is that they learn. The process of learning and what is learned are impacted by a number of factors, both cognitive and social; that is, humans are boundedly rational. Cognitive and social limitations interact, making it difficult to reason about how to provide information to impact what humans know, believe, and do. Herein, we use a multi-agent dynamic-network simulation system, Construct, to conduct such reasoning. In particular, we ask, What media should be used to provide information to most impact what people know, believe, and do, given diverse social structures? All simulated agents are boundedly rational both at the cognitive and social level, and so are subject to factors such as literacy, education, and the breadth of their social network. We find that there is no one most effective intervention; rather, to be effective, messages and the media used to spread the message need to be selected for the population being addressed. Typically, a multimedia campaign is critical.


The Association of Fractalkine Receptor (T280M) Polymorphism in the Pathogenesis of Acute Coronary Syndrome in the Egyptian Population.
Fractalkine (FKN) in its free and membrane bound-forms and its receptor CX3CR1are reported to have an atherosclerotic effect. The relationship of Single Nucleotide Polymorphisms (SNPs) in FKN and CX3CR1genes with the Coronary Artery Disease (CAD) risk showed conflicting results in different populations. The aim of this study was to investigate the influence of CX3CR1 threonine 280 methionine (T280M) polymorphism in the predisposition of Acute Coronary Syndrome (ACS) in Egyptians. 200 Egyptian subjects were recruited for the study. They were divided into 100 ACS patients and 100 healthy controls. Genotyping of CX3CR1 T280M was performed using a Polymerase Chain Reaction-restriction Fragment Length Polymorphism (PCR-RFLP). Serum FKN was assayed by Enzyme - Linked - Immuno- Sorbent-Assay (ELISA). T and M allele frequencies for CX3CR1gene were not significantly different between ACS and Controls (p=0.76). Moreover, none of the genotypes had an atheroprotective effect. Serum analysis showed higher levels of FKN in ACS patients (p=0.041). FKN levels were not significantly different among genotypes of control and ACS groups (p=0.34) and (p=0.38) respectively. This study shows that CX3CR1 T280M polymorphism does not affect the incidence of ACS the Egyptian population. Moreover, none of the genotypes were associated with higher FKN levels.


Broadband focused waves with compensated spatial dispersion: transverse versus axial balance.
We determine the constraints an ABCD optical system must verify to achieve, at the focal region, broadband waves with compensated spatial dispersion either along the optical axis, called on-axis isodiffracting fields, or in the lateral direction, here named in-plane isodiffracting beams. An optical configuration is identified for generating both types of achromatic broadband focused wave fields. An experimental verification is also provided.


Src-dependent Syk activation controls CD69-mediated signaling and function on human NK cells.
CD69 C-type lectin receptor represents a functional triggering molecule on activated NK cells, capable of directing their natural killing function. The receptor-proximal signaling pathways activated by CD69 cross-linking and involved in CD69-mediated cytotoxic activity are still poorly understood. Here we show that CD69 engagement leads to the rapid and selective activation of the tyrosine kinase Syk, but not of the closely related member of the same family, ZAP70, in IL-2-activated human NK cells. Our results indicate the requirement for Src family kinases in the CD69-triggered activation of Syk and suggest a role for Lck in this event. We also demonstrate that Syk and Src family tyrosine kinases control the CD69-triggered tyrosine phosphorylation and activation of phospholipase Cgamma2 and the Rho family-specific exchange factor Vav1 and are responsible for CD69-triggered cytotoxicity of activated NK cells. The same CD69-activated signaling pathways are also observed in an RBL transfectant clone, constitutively expressing the receptor. These data demonstrate for the first time that the CD69 receptor functionally couples to the activation of Src family tyrosine kinases, which, by inducing Syk activation, initiate downstream signaling pathways and regulate CD69-triggered functions on human NK cells.


Impact of depressive symptoms on visceral sensitivity among patients with different subtypes of irritable bowel syndrome.
The etiology of irritable bowel syndrome (IBS) is complex and multifaceted. Psychosocial factors play a role in such a process. Several reports suggest that IBS patients have increased psychopathology scores as compared with population controls. The influence of depressive symptoms on rectal sensitivity thresholds vary across different studies. The influence of predominant bowel habits on rectal sensitivity thresholds as determined by barostat-based investigations is not well established. The present report aimed to investigate the influence of depressive symptoms on rectal sensitivity in different subtypes of IBS patients (diarrhea/constipation-predominant vs. alternating subtypes). Depressive symptoms correlated well with first pain sensitivity threshold in alternating patients (n = 8; [rho] = -0.77; p = 0.02) but not in diarrhea/constipation predominant symptoms (n = 11; [rho] = -0.44; p = 0.27). These data suggest that depressive symptoms might impact pain thresholds differently according to the subtype of IBS.


Macrocyclic peptides self-assemble into robust vesicles with molecular recognition capabilities.
In this study, we developed macrocyclic peptide building blocks that formed self-assembled peptide vesicles with molecular recognition capabilities. Macrocyclic peptides were significantly different from conventional amphiphiles, in that they could self-assemble into vesicles at very high hydrophilic-to-total mass ratios. The flexibility of the hydrophobic self-assembly segment was critical for vesicle formation. The unique features of this peptide vesicle system include a homogeneous size distribution, unusually small size, and robust structural and thermal stability. The peptide vesicles successfully entrapped a hydrophilic model drug, released the payload very slowly, and were internalized by cells in a highly efficient manner. Moreover, the peptide vesicles exhibited molecular recognition capabilities, in that they selectively bound to target RNA through surface-displayed peptides. This study demonstrates that self-assembled peptide vesicles can be used as strong intracellular delivery vehicles that recognize specific biomacromolecular targets.


Drug-resistant breast cancer cells frequently retain expression of a functional wild-type p53 protein.
Abnormalities in the p53 tumor suppressor gene have been shown to affect cellular processes related to cell cycle control and gene amplification. In this study we compare the status and function of wild-type p53 in MCF-7 breast cancer cells with sublines selected for resistance to chemotherapeutic agents having different mechanisms of action. Sublines that were resistant to melphalan, pyrazafurin, mitoxantrone, etoposide and PALA all retained expression of wild-type p53. Methotrexate-resistant MCF-7 cells were unusual heterozygotes that expressed a wild-type and dominant, in-frame p53 deletion mutant and the doxorubicin-resistant cells expressed only mutant p53. Analysis of the G1 checkpoint after treatment with ionizing radiation revealed that the pyrazafurin-, melphalan- and mitoxantrone-resistant cells arrested strongly in G1. The etoposide- and PALA-resistant cells had an intermediate G1 arrest phenotype and the methotrexate- and doxorubicin-resistant cells had a minimal G1 arrest phenotype. mRNA and protein analyses of downstream effector genes, including P21CIP1/Waf1, mdm2, Gadd 45 and the retinoblastoma protein, did not entirely differentiate sublines having a strong versus intermediate G1 arrest phenotype. Neither the p53 status nor the strength of the G1 arrest could be correlated with cell survival after ionizing radiation. When drug-sensitive MCF-7 cells were treated with the same chemotherapeutic agents, p53 and p21CIP1/Waf1 levels increased between 2- and 14-fold. Together these data suggest that other cellular factors likely play a role in overcoming the inhibitory effects of ionizing radiation on p53 in drug-resistant breast cancer cells.


A comparison of pathological methods of measuring lung cancer volume.
To determine which of several pathological methods of measuring lung cancer volume compared most favourably with the gold standard. Three pathological methods were used on 54 resected lung cancers: (1) measuring the maximum dimension and assuming a spherical shape; (2) measuring three dimensions and assuming an ellipsoidal shape; and (3) deriving the volume from the area of tumour on sequential 1 cm slices using a photocopier and an image analysis system. The gold standard was obtained from the area of whole mount tumour sections on sequential 0.1 cm slices of eight cancers. Volumes derived from 1 cm lung slices gave results closest to our gold standard but assuming tumours were ellipsoidal was only a slightly less accurate and less time consuming method. Assuming cancers were spherical resulted in gross overestimation of the tumour volumes. For practical purposes, it is reasonable to measure three dimensions of a lung tumour at sectioning and calculate the volume using the formula for an ellipsoid (V = 4/3 pi d.e.f, where d, e and f are the semi-axes).


Breakthrough cancer pain: review of prevalence, characteristics and management.
Breakthrough pain has been associated with a reduced likelihood of adequate pain control. Despite the large and variable incidence of this phenomenon due to varied definitions of this type of pain, only a few studies have been conducted to assess and effectively treat breakthrough pain though the importance of managing breakthrough pain has been acknowledged by all. A large number of drugs from various classes and novel methods of administration like nasal and transmucosal buccal route, as in the case of fentanyl, have been used in these studies to manage this type of pain. A drug is needed with a quick onset of action and optimal duration that matches the characteristics of breakthrough pain. Some steps have been taken in earlier studies which used nasal formulation of fentanyl as it was found to achieve adequate and quick pain relief. However, further studies are required to confirm this so that in the future we can have as effective protocols for managing breakthrough pain as we have today for managing persistent pain as given by the World Health Organization.


Repositioning techniques for malpositioned neonatal peripherally inserted central catheters.
The utilization of Peripherally Inserted Central Catheters (PICCs) has grown to become a common practice in neonatal care. Proper placement of the catheter in the superior vena cava (SVC) is essential to support safe delivery of care and minimize complications. This article describes three cases of patients whose catheters were malpositioned on initial placement or while in situ, that were successfully guided to appropriate tip location in the superior vena cava by implementing a combination of non-invasive catheter repositioning techniques.


Physiological concentrations of insulin and T3 stimulate 3T3-L1 adipocyte acyl-CoA synthetase gene transcription.
Acyl-CoAsynthetase (ACS) is a key gene for cellular utilization of long-chain fatty acids. We characterized its regulation by physiological concentrations of insulin that acutely regulate metabolism. Our results demonstrate that subnanomolar insulin rapidly and maximally stimulates ACS gene transcription in the absence of protein synthesis; 0.5 nM insulin produced a 2.3 +/- 0.1-fold increase in ACS mRNA levels and induced ACS gene transcription 2.4 +/- 0.3-fold. The insulin sensitivity of ACS was compared with lipoprotein lipase (LPL) and stearoyl-CoA desaturase-1 (SCD-1), which were both less sensitive to insulin. Physiological triiodothyronine (10 nm) also induced ACS mRNA 2.4 +/- 0.1-fold and gene transcription 2.8 +/- 0.3-fold and coordinately induced LPL and SCD-1 mRNA and gene transcription. Because insulin and adenosine 3',5'-cyclic monophosphate often regulate genes involved in lipid and carbohydrate metabolism in a reciprocal manner, we evaluated effects of 1-methyl-3-isobutylxanthine (MIX).ACS mRNA levels were strongly downregulated by MIX in a dose-dependent manner, and ACS gene transcription inhibited in a coordinate manner with LPL and SCD-1. These data demonstrate a uniquely sensitive pattern of stimulation of ACS gene transcription by insulin with reciprocal regulation by MIX, and they suggest a significant role for ACS as a tightly regulated "gatekeeper" gene participating in the control of adipocyte metabolism.


Assessment of Heart Rate Monitoring During Exercise With Smart Wristbands and a Heart Rhythm Patch: Validation and Comparison Study.
The integration of wearable devices into fitness routines, particularly in military settings, necessitates a rigorous assessment of their accuracy. This study evaluates the precision of heart rate measurements by locally manufactured wristbands, increasingly used in military academies, to inform future device selection for military training activities. This research aims to assess the reliability of heart rate monitoring in chest straps versus wearable wristbands. Data on heart rate and acceleration were collected using the Q-Band Q-69 smart wristband (Mobile Action Technology Inc) and compared against the Zephyr Bioharness standard measuring device. The Lin concordance correlation coefficient, Pearson product moment correlation coefficient, and intraclass correlation coefficient were used for reliability analysis. Participants from a Northern Taiwanese medical school were enrolled (January 1-June 31, 2021). The Q-Band Q-69 demonstrated that the mean absolute percentage error (MAPE) of women was observed to be 13.35 (SD 13.47). Comparatively, men exhibited a lower MAPE of 8.54 (SD 10.49). The walking state MAPE was 7.79 for women and 10.65 for men. The wristband's accuracy generally remained below 10% MAPE in other activities. Pearson product moment correlation coefficient analysis indicated gender-based performance differences, with overall coefficients of 0.625 for women and 0.808 for men, varying across walking, running, and cooldown phases. This study highlights significant gender and activity-dependent variations in the accuracy of the MobileAction Q-Band Q-69 smart wristband. Reduced accuracy was notably observed during running. Occasional extreme errors point to the necessity of caution in relying on such devices for exercise monitoring. The findings emphasize the limitations and potential inaccuracies of wearable technology, especially in high-intensity physical activities.


Intermittent gastric outlet obstruction caused by a prolapsing antral gastric polyp.
Most gastric polyps have an asymptomatic presentation and are an incidental finding on upper endoscopy. Symptomatic presentations can range from an ulcerated polyp leading to anemia and occult bleed to complete gastric outlet obstruction. We report a case of an 89-year-old woman who presented with postprandial nausea and early satiety. Her upper endoscopy revealed a 2 cm pedunculated hyperplastic polyp arising from the antrum of the stomach which was seen prolapsing into the pylorus causing intermittent gastric outlet obstruction. In the present report, we statistically analyzed 39 prolapsing gastric polyps previously reported in the English literature and demonstrate the current utility of monopolar snare polypectomy in establishing a histological diagnosis while offering simultaneous treatment. Additionally, we review the literature for the management of all hyperplastic gastric polyps in relation to advancements in digestive endoscopy.


Spatial regulation of membrane fusion controlled by modification of phosphoinositides.
Membrane fusion plays a central role in many cell processes from vesicular transport to nuclear envelope reconstitution at mitosis but the mechanisms that underlie fusion of natural membranes are not well understood. Studies with synthetic membranes and theoretical considerations indicate that accumulation of lipids characterised by negative curvature such as diacylglycerol (DAG) facilitate fusion. However, the specific role of lipids in membrane fusion of natural membranes is not well established. Nuclear envelope (NE) assembly was used as a model for membrane fusion. A natural membrane population highly enriched in the enzyme and substrate needed to produce DAG has been isolated and is required for fusions leading to nuclear envelope formation, although it contributes only a small amount of the membrane eventually incorporated into the NE. It was postulated to initiate and regulate membrane fusion. Here we use a multidisciplinary approach including subcellular membrane purification, fluorescence spectroscopy and Förster resonance energy transfer (FRET)/two-photon fluorescence lifetime imaging microscopy (FLIM) to demonstrate that initiation of vesicle fusion arises from two unique sites where these vesicles bind to chromatin. Fusion is subsequently propagated to the endoplasmic reticulum-derived membranes that make up the bulk of the NE to ultimately enclose the chromatin. We show how initiation of multiple vesicle fusions can be controlled by localised production of DAG and propagated bidirectionally. Phospholipase C (PLCgamma), GTP hydrolysis and (phosphatidylinsositol-(4,5)-bisphosphate (PtdIns(4,5)P(2)) are required for the latter process. We discuss the general implications of membrane fusion regulation and spatial control utilising such a mechanism.


Operando Reactor-Cell with Simultaneous Transmission FTIR and Raman Characterization (IRRaman) for the Study of Gas-Phase Reactions with Solid Catalysts.
Raman and transmission FTIR spectroscopic techniques have been coupled in a new homemade reactor-cell designed in a joint CSIC-LCS collaboration. The setup is easily adapted to any FTIR and fiber-coupled Raman spectrometers and gas analysis techniques. It allows for simultaneous operando FTIR and Raman spectroscopic measurement, which provide complementary characterization of adsorbed species, reaction intermediates, and structural properties of the catalyst. This system was validated with the study of vanadium-based catalysts during propane oxydehydrogenation (ODH). The combined use of both spectroscopies with gas analysis techniques to measure the activity contributes to the understanding of propane ODH and the identification of the role of different oxygen species bound to vanadium sites. For example, the simultaneous characterization of the catalyst under the same conditions by IR and Raman confirms that the V═O mode has the same frequency in both spectroscopies and that bridging oxygen sites (V-O-V, V-O-Zr) present higher activity than terminal V═O bonds. These results demonstrate the high potential of the new simultaneous transmission IR-Raman operando rig to correlate the activity and the structure of catalysts, thus assisting the rational design of catalytic processes.


Accumulation of Ag(0) Single Atoms at Water/Mineral Interfaces during Sunlight-Induced Reduction of Ionic Ag by Phenolic DOM.
Silver (Ag) undergoes a complex and dynamic Ag+/Ag0 cycle under environmental conditions. The Ag+ → Ag nanoparticles (AgNPs) transformation due to the combined actions of sunlight, O2, and dissolved organic matter has been a well-known environmental phenomenon. In this study, we indicate that this process may be accompanied by a pronounced accumulation of Ag(0) single atoms (Ag-SAs) on the minerals' surfaces. According to spherical aberration-corrected scanning transmission electron microscopy and high-energy-resolution X-ray adsorption fine structure analyses, humic acid (HA) and phenol (PhOH) can induce Ag-SAs accumulation, whereas oxalic acid causes only AgNPs deposition. Ag-SAs account for more than 20 wt % of total Ag(0) on the γ-Al2O3 surfaces during HA- and PhOH-mediated photolysis processes. HA also causes Ag-SAs to accumulate on two other prevalent soil minerals, SiO2 and Fe2O3, and the fractions of Ag-SAs are about 15 wt %. Our mechanism studies suggest that a phenolic molecule acts as a reducing agent of Ag+ and a stabilizer of Ag-SAs, protecting Ag-SAs against autocatalytic nucleation.


Are cannabis prevalence estimates comparable across countries and regions? A cross-cultural validation using search engine query data.
Prevalence estimation of cannabis use is usually based on self-report data. Although there is evidence on the reliability of this data source, its cross-cultural validity is still a major concern. External objective criteria are needed for this purpose. In this study, cannabis-related search engine query data are used as an external criterion. Data on cannabis use were taken from the 2007 European School Survey Project on Alcohol and Other Drugs (ESPAD). Provincial data came from three Italian nation-wide studies using the same methodology (2006-2008; ESPAD-Italia). Information on cannabis-related search engine query data was based on Google search volume indices (GSI). (1) Reliability analysis was conducted for GSI. (2) Latent measurement models of "true" cannabis prevalence were tested using perceived availability, web-based cannabis searches and self-reported prevalence as indicators. (3) Structure models were set up to test the influences of response tendencies and geographical position (latitude, longitude). In order to test the stability of the models, analyses were conducted on country level (Europe, US) and on provincial level in Italy. Cannabis-related GSI were found to be highly reliable and constant over time. The overall measurement model was highly significant in both data sets. On country level, no significant effects of response bias indicators and geographical position on perceived availability, web-based cannabis searches and self-reported prevalence were found. On provincial level, latitude had a significant positive effect on availability indicating that perceived availability of cannabis in northern Italy was higher than expected from the other indicators. Although GSI showed weaker associations with cannabis use than perceived availability, the findings underline the external validity and usefulness of search engine query data as external criteria. The findings suggest an acceptable relative comparability of national (provincial) prevalence estimates of cannabis use that are based on a common survey methodology. Search engine query data are a too weak indicator to base prevalence estimations on this source only, but in combination with other sources (waste water analysis, sales of cigarette paper) they may provide satisfactory estimates.


From accuracy to patient outcome and cost-effectiveness evaluations of diagnostic tests and biomarkers: an exemplary modelling study.
Proper evaluation of new diagnostic tests is required to reduce overutilization and to limit potential negative health effects and costs related to testing. A decision analytic modelling approach may be worthwhile when a diagnostic randomized controlled trial is not feasible. We demonstrate this by assessing the cost-effectiveness of modified transesophageal echocardiography (TEE) compared with manual palpation for the detection of atherosclerosis in the ascending aorta. Based on a previous diagnostic accuracy study, actual Dutch reimbursement data, and evidence from literature we developed a Markov decision analytic model. Cost-effectiveness of modified TEE was assessed for a life time horizon and a health care perspective. Prevalence rates of atherosclerosis were age-dependent and low as well as high rates were applied. Probabilistic sensitivity analysis was applied. The model synthesized all available evidence on the risk of stroke in cardiac surgery patients. The modified TEE strategy consistently resulted in more adapted surgical procedures and, hence, a lower risk of stroke and a slightly higher number of life-years. With 10% prevalence of atherosclerosis the incremental cost-effectiveness ratio was € 4,651 and € 481 per quality-adjusted life year in 55-year-old men and women, respectively. In all patients aged 65 years or older the modified TEE strategy was cost saving and resulted in additional health benefits. Decision analytic modelling to assess the cost-effectiveness of a new diagnostic test based on characteristics, costs and effects of the test itself and of the subsequent treatment options is both feasible and valuable. Our case study on modified TEE suggests that it may reduce the risk of stroke in cardiac surgery patients older than 55 years at acceptable cost-effectiveness levels.


Fine particulate matter and cardiovascular disease: Comparison of assessment methods for long-term exposure.
Adverse cardiovascular events have been linked with PM2.5 exposure obtained primarily from air quality monitors, which rarely co-locate with participant residences. Modeled PM2.5 predictions at finer resolution may more accurately predict residential exposure; however few studies have compared results across different exposure assessment methods. We utilized a cohort of 5679 patients who had undergone a cardiac catheterization between 2002-2009 and resided in NC. Exposure to PM2.5 for the year prior to catheterization was estimated using data from air quality monitors (AQS), Community Multiscale Air Quality (CMAQ) fused models at the census tract and 12km spatial resolutions, and satellite-based models at 10km and 1km resolutions. Case status was either a coronary artery disease (CAD) index >23 or a recent myocardial infarction (MI). Logistic regression was used to model odds of having CAD or an MI with each 1-unit (μg/m3) increase in PM2.5, adjusting for sex, race, smoking status, socioeconomic status, and urban/rural status. We found that the elevated odds for CAD>23 and MI were nearly equivalent for all exposure assessment methods. One difference was that data from AQS and the census tract CMAQ showed a rural/urban difference in relative risk, which was not apparent with the satellite or 12km-CMAQ models. Long-term air pollution exposure was associated with coronary artery disease for both modeled and monitored data.


A super-family of genes coding for secreted salivary gland proteins from the Hessian fly, Mayetiola destructor.
We have previously characterized a gene coding for the secreted-salivary-gland-protein 11A1 (SSGP-11A1) from the Hessian fly, Mayetiola destructor (Say) (Diptera Cecidomyiidae). Here we report the cloning and characterization of three new genes coding for proteins designated SSGP-11B1, SSGP-11C1, and SSGP-11C2, and their relationship with the SSGP-11A1-encoding gene. Based on their structural conservation, similar regulation, and clustered genomic organization, we conclude that the four genes represent a gene superfamily, designated SSGP-11, which originated from a common ancestor. Cloning, Southern blot and in situ hybridization data suggest that each of theSSGP-11 families has multiple members that cluster within short chromosome regions. The presence of a secretion signal peptide, the exclusive expression in the larval stage, and the clustered genomic organization indicate that this superfamily might be important for Hessian fly virulence/avirulence.


Afatinib versus cisplatin plus gemcitabine for first-line treatment of Asian patients with advanced non-small-cell lung cancer harbouring EGFR mutations (LUX-Lung 6): an open-label, randomised phase 3 trial.
Afatinib-an oral irreversible ErbB family blocker-improves progression-free survival compared with pemetrexed and cisplatin for first-line treatment of patients with EGFR mutation-positive advanced non-small-cell lung cancer (NSCLC). We compared afatinib with gemcitabine and cisplatin-a chemotherapy regimen widely used in Asia-for first-line treatment of Asian patients with EGFR mutation-positive advanced NSCLC. This open-label, randomised phase 3 trial was done at 36 centres in China, Thailand, and South Korea. After central testing for EGFR mutations, treatment-naive patients (stage IIIB or IV cancer [American Joint Committee on Cancer version 6], performance status 0-1) were randomly assigned (2:1) to receive either oral afatinib (40 mg per day) or intravenous gemcitabine 1000 mg/m(2) on day 1 and day 8 plus cisplatin 75 mg/m(2) on day 1 of a 3-week schedule for up to six cycles. Randomisation was done centrally with a random number-generating system and an interactive internet and voice-response system. Randomisation was stratified by EGFR mutation (Leu858Arg, exon 19 deletions, or other; block size three). Clinicians and patients were not masked to treatment assignment, but the independent central imaging review group were. Treatment continued until disease progression, intolerable toxic effects, or withdrawal of consent. The primary endpoint was progression-free survival assessed by independent central review (intention-to-treat population). This study is registered with ClinicalTrials.gov, NCT01121393. 910 patients were screened and 364 were randomly assigned (242 to afatinib, 122 to gemcitabine and cisplatin). Median progression-free survival was significantly longer in the afatinib group (11·0 months, 95% CI 9·7-13·7) than in the gemcitabine and cisplatin group (5·6 months, 5·1-6·7; hazard ratio 0·28, 95% CI 0·20-0·39; p<0·0001). The most common treatment-related grade 3 or 4 adverse events in the afatinib group were rash or acne (35 [14·6%] of 239 patients), diarrhoea (13 [5·4%]), and stomatitis or mucositis (13 [5·4%]), compared with neutropenia (30 [26·5%] of 113 patients), vomiting (22 [19·5%]), and leucopenia (17 [15·0%]) in the gemcitabine and cisplatin group. Treatment-related serious adverse events occurred in 15 (6·3%) patients in the afatinib group and nine (8·0%) patients in the gemcitabine and cisplatin group. First-line afatinib significantly improves progression-free survival with a tolerable and manageable safety profile in Asian patients with EGFR mutation-positive advanced lung NSCLC. Afatinib should be considered as a first-line treatment option for this patient population. Boehringer Ingelheim.


Significant build-up of soil organic carbon under climate-smart conservation farming in Sub-Saharan Acrisols.
Conservation farming (CF) involving minimum tillage, mulching and crop rotation may offer climate change adaptation and mitigation benefits. However, reported effects of CF, as applied by smallholders, on storage of soil organic carbon (SOC) and soil fertility in Sub-Saharan Africa differ considerably between studies. This is partly due to differences in management practice, soil type and adoption level between individual farmers. Where CF involves planting basins, year-to-year changes in position of basins make SOC stock estimates more uncertain. Here we assess the difference in SOC build-up and soil quality between inside planting basins (receiving inputs of lime and fertilizer; basins opened each year) and outside planting basins (no soil disturbance or inputs other than residues) under hand-hoe tilled CF in an Acrisol at Mkushi, Zambia. Seven years of strict CF husbandry significantly improved soil quality inside planting basins as compared with outside basins. Significant effects were found for SOC concentration (0.74 ± 0.06% vs. 0.57 ± 0.08%), SOC stock (20.1 ± 2.0 vs. 16.4 ± 2.6 t ha-1, 0-20 cm), soil pH (6.3 ± 0.2 vs. 4.95± 0.4) and cation exchange capacity (3.8 ± 0.7 vs. 1.6 ± 0.4 cmolc kg-1). As planting basins only occupy 9.3% of the field, the absolute rate of increase in SOC, compared with outside basins, was 0.05 t C ha-1 yr-1. This corresponds to an overall relative increase of 2.95‰ SOC yr-1 in the upper 20 cm of the soil. Also, hot water extractable carbon (HWEC), a proxy for labile organic matter, and potential nitrification rates were consistently greater inside than outside basins. The significant increase in quantity and quality of SOC may be due to increased inputs of roots, due to favorable conditions for plant growth through input of fertilizer and lime, along with increased rainwater infiltration in the basins.


Efficacy and safety of once versus twice daily administration of methimazole in cats with hyperthyroidism.
To determine whether once daily administration of methimazole was as effective and safe as twice daily administration in cats with hyperthyroidism. Randomized, nonblinded, clinical trial. 40 cats with newly diagnosed hyperthyroidism. Cats were randomly assigned to receive 5 mg of methimazole, PO, once daily (n = 25) or 2.5 mg of methimazole, PO, twice daily (15). A complete physical examination, including measurement of body weight; CBC; serum biochemical analyses, including measurement of serum thyroxine concentration; and urinalysis were performed, and blood pressure was measured before and 2 and 4 weeks after initiation of treatment. Serum thyroxine concentration was significantly higher in cats given methimazole once daily, compared with cats given methimazole twice daily, 2 weeks (3.7 vs 2.0 micro +/- g/dL) and 4 weeks (3.2 vs 1.7 microg/dL) after initiation of treatment. In addition, the proportion of cats that were euthyroid after 2 weeks of treatment was lower for cats receiving methimazole once daily (54%) than for cats receiving methimazole twice daily (87%). Percentages of cats with adverse effects (primarily gastrointestinal tract upset and facial pruritus) were not significantly different between groups. Results suggest that once daily administration of methimazole was not as effective as twice daily administration in cats with hyperthyroidism and cannot be recommended for routine use.


Daily physical activity in low-risk extremely low birth weight preterm infants: positive impact on bone mineral density and anthropometric measurements.
The aim of this study was to evaluate the changes in bone mineral density and anthropometric indices of extremely low birth weight (ELBW) preterm infants undergoing daily physical activity. Twenty-eight low-risk ELBW preterm infants (intervention group = 14, control group = 14) with a birth weight of under 1000 g and gestational age of 26-32 weeks were recruited. Preterm infants in the control group were provided with standard nursing care, whereas those in the intervention group received a daily physical activity programme in addition to standard nursing care during the study. Before and after the study, anthropometric indices and tibial speed of sound (SOS) values were measured. In this study, the tibial SOS values were found to be increased in the intervention group (+111.14 m/s) (p = 0.001) and decreased in the control group (-58.21 m/s) (p = 0.030). The increase in percentage of body weight, height, and tibial length of infants in the intervention group was significantly higher than in the control group (respectively; p = 0.002, p = 0,015, p < 0.001). A daily physical activity programme increased body weight, height, tibial length, and SOS values of the tibia in ELBW preterm infants.


An insight into the catalytic activity of palladium Schiff-base complexes towards the Heck coupling reaction: routes via encapsulation in zeolite-Y.
A small set of palladium Schiff-base complexes were synthesized and entrapped in the supercage of zeolite-Y. All these novel complexes in both states were systematically characterized with the help of different characterization tools like XRD, SEM-EDS, thermal analysis, XPS, IR, electronic spectroscopic and theoretical studies. These systems were thoroughly studied for their catalytic activities towards the Heck coupling reaction between bromobenzene and styrene. The aim was to meticulously compare the performance of the homogeneous catalysts, i.e., neat palladium Schiff-base complexes with that of their heterogeneous encapsulated analogs. The experimental as well as theoretical electronic structure studies suggested significant structural modification of the guest Pd(ii)-Schiff-base complexes after encapsulation in zeolite Y. These complexes manifested modified catalytic activities towards the Heck coupling reaction. The theoretical studies reinforced the correlation between the modified catalytic properties and structural alteration of these complexes on encapsulation. These heterogeneous catalysts essentially demonstrated the benefits of easy separation and reusability as compared to the homogeneous analogues.


Characterization of intermolecular G-quadruplex formation over intramolecular G-triplex for DNA containing three G-tracts.
G-triplex (G3) has been recognized as a popular intermediate during the folding of G-quadruplex (G4). This has raised interest to anticipate the ultimate formation of G3 by shortening the G4-forming oligonucleotides with the remaining three G-tracts. Some G3 structures have been validated and their stability has been found to be affected by the loop sequences similar to G4s. In this work, however, we first found that an intermolecular parallel G4 structure was preferred in K+ for the oligonucleotide 5'-TGGGTAGGGCGGG-3' (DZ3) containing only three G-tracts. We screened auramine O (AO) as the appropriate fluorophore with a molecular rotor feature to target this G4 structure. AO bound with DZ3 in a 1 : 4 ratio, as confirmed by isothermal titration calorimetry experiments, suggesting the formation of a tetramolecular G4 structure (4erG4). The excimer emission from the labelled pyrene and the DNA melting behavior at various pHs in the presence of Ag+ proved the formation of the 4erG4 structure rather than the prevalent intramolecular G3 folding. This work demonstrates that one should be cautious while putatively predicting a G3 structure from an oligonucleotide containing three G-tracts.


[Scoliosis: review of types, aetiology, diagnostics, and treatment 2].
In congenital scoliosis, an anatomical spine deformity arose during embryonic vertebral development which caused a deformity. Approximately 38-55% of these deformities appear as a syndrome. Prognosis and preferred treatment depend on the type offailure and the severity of the deformity, but surgical correction is often necessary. Neuromuscular scoliosis generally arises at a young age, is often rapidly progressive, and the magnitude of the curve can also progress even in adulthood. The prognosis of the curve together with the symptoms and prognosis of the associated neuromuscular condition determine the preferred treatment. The goal of surgical treatment is to improve the patient's quality of life. Two types of adult scoliosis occur: adult idiopathic scoliosis and adult degenerative scoliosis. The surgical principles for adult idiopathic scoliosis are almost identical to those for adolescent idiopathic scoliosis. In degenerative scoliosis, there is almost always a shorter fusion length. In the case of neurologic impairments, decompression of the spinal cord or nerve roots should be carried out.


Evaluation of progesterone levels in feces of captive reticulated giraffe (Giraffa camelopardalis reticulata).
Fresh fecal samples were collected from seven adult female reticulated giraffe (Giraffa camelopardalis reticulata). Samples were collected for several weeks before, during, and for a few weeks after gestation. Fecal samples were analyzed for progesterone levels by radioimmunoassay. There were significant differences in progesterone levels between pregestational and gestational samples and between gestational and postgestational samples. These results demonstrate that fecal progesterone levels are useful in determining pregnant versus nonpregnant reticulated giraffe.


Stopping versus continuing acetylsalicylic acid before coronary artery bypass surgery: A systematic review and meta-analysis of 14 randomized controlled trials with 4499 patients.
This study aimed to evaluate the efficacy and safety of continuing versus stopping aspirin [acetylsalicylic acid (ASA)] preoperatively in patients undergoing coronary artery bypass graft surgery. MEDLINE, EMBASE, CENTRAL/Cochrane Controlled Trials Register (CCTR), ClinicalTrials.gov, Scientific Electronic Library Online (SciELO), Literatura Latino Americana em Ciências da Saúde (LILACS), Google Scholar and reference lists of relevant articles were searched for randomized controlled trials that reported efficacy outcomes of myocardial infarction and mortality, and safety outcomes of blood loss, packed red blood cell transfusion and surgical re-exploration were compared between groups. Fourteen studies fulfilled our eligibility criteria and included a total of 4499 patients (2329 for 'continuing ASA' and 2170 for 'stopping ASA'). In the pooled analysis, continuing aspirin therapy did not reduce the risk of myocardial infarction [risk ratio 0.834, 95% confidence interval (CI) 0.688-1.010; P = 0.063] or operative mortality (risk ratio 1.384, 95% CI 0.727-2.636; P = 0.323). Preoperative ASA increased postoperative chest tube drainage (mean difference 143 ml, 95% CI 39-248 ml; P = 0.007) and packed red blood cell transfusion (mean difference 142 ml, 95% CI 55-228; P = 0.001) but did not increase the risk of surgical re-exploration (risk ratio 1.316, 95% CI 0.910-1.905; P = 0.145). This meta-analysis found no statistically significant difference regarding the risk of operative mortality and myocardial infarction between the 'continuing ASA' and 'stopping ASA' strategies. On the other hand, the mean volume of blood loss and packed red blood cell transfusion was higher in the 'continuing ASA' group, but this finding did not translate into higher risk of reoperation for bleeding.


[Clinical, biochemical and histological indicators of Del Nido cardioplegia efficacy in patients with severe myocardial hypertrophy].
To evaluate the effectiveness of two cardioplegia techniques in patients with severe myocardial hypertrophy undergoing septal myectomy or aortic valve replacement. A comparative pilot prospective single-center randomized study included 46 patients between 2022 and 2023. Patients were randomized into 2 groups: Del Nido (n=23) and Custodiol (n=23). We analyzed perioperative echocardiography data, troponin I at several time points, perioperative complications and histological data. Both groups were comparable in time of myocardial ischemia, cardiopulmonary bypass, duration of anesthesia and surgery (p>0.05). The maximum ischemia time in the Del Nido group was 84 min. The same group showed significantly higher percentage of spontaneous rhythm recovery (65.2% vs. 30%, p=0.008). None patient required mechanical support, high-dose inotropes or vasopressors. Troponin I in 2 hours after cardiopulmonary bypass (p=0.415), 12 (p=0.528) and 24 hours after admission to the intensive care unit (p=0.281) were similar in both groups. No significant difference was found in ventilation time, ICU- and hospital-stay. Del Nido cardioplegia has some advantages compared to Custodiol and does not lead to perioperative complications in case of aortic cross-clamping time <90 min in patients with myocardial hypertrophy.


Effect of hyperbaric oxygen therapy in rats with subtotal splenectomy preserving the inferior pole.
To evaluate the effect of hyperbaric oxygen therapy on the survival and weight of rats submitted to subtotal splenectomy and on the viability and growth of the inferior pole. Forty Wistar rats underwent subtotal splenectomy with preservation of the inferior pole and distributed into two groups: Group A (n=20) - not treated with hyperbaric oxygen, Group B (n=20) – treated with hyperbaric oxygen. These groups were divided into two subgroups of 10 animals each, according to the time of euthanasia, 15th and 45th days. The survival and weight of the animals were recorded. The inferior pole was measured, weighed and morphologically analyzed. All animals survived. The weight of the animals increased in all subgroups, but decreased on the 10th day in the subgroups treated with hyperbaric oxygen (p<0.001). The viability of the inferior pole was more evident in animals treated on the 15th day, but did not differ on the 45th day. The growth of the inferior pole has not occurred on the 15th day but on the 45th day after surgery in untreated animals (p<0.01) and treated animals (p<0.05). Vascular and cellular increase in treated animals was significantly higher than in untreated ones. Hyperbaric oxygen therapy did not affect the survival of animals but reduced their weight. It improved the viability of the inferior splenic pole, but did not interfere with their growth.


Psychopathic-like traits in detained adolescents: clinical usefulness of self-report.
Studies have demonstrated that self-report tools can be used to reliably and validly examine psychopathic-like traits in adolescents. However, it is unclear if self-report instruments are still reliable and valid when confidentiality cannot be guaranteed, such as during routine assessments in juvenile detention centres. To address this issue, the current study used data from the routine mental health screening of 365 detained male adolescents (12-18 years) in two juvenile detention centres. With the intention of gaining insight in the clinical usefulness of self-reported psychopathic-like traits, we examined relations known from literature with emotional and behavioural features. Self-reported psychopathic-like traits, measured by the Youth Psychopathic Traits Inventory-Short version (YPI-S), were uniquely associated with substance abuse, anger/irritability, conduct problems and hyperactivity, but not with internalizing problems. YPI-S-dimensions showed several specific relationships with variables of interest. For example, only the callous unemotional dimension was negatively related with prosocial behaviour and only the behavioural dimension was positively related with hyperactivity. In conclusion, self-reported psychopathic-like traits showed expected relations with relevant variables. These findings suggest that self-report can be used to identify detained youths with high levels of psychopathic-like traits outside a research context, thus, even when anonymity and confidentiality are not guaranteed.


Mussel-inspired direct immobilization of nanoparticles and application for oil-water separation.
Immobilization of various nanoparticles onto complex 2D or 3D macroscopic surface is an important issue for nanotechnology, but the challenge remains to explore a facile, general and environmentally friendly method for achieving this goal. Taking inspiration from the adhesion of marine mussels, we reported here that oxide nanoparticles of different compositions and sizes were directly and robustly anchored on the surface of monolithic foams ranging from polymer to metals in an aqueous solution of dopamine. The effective immobilization of the nanoparticles was strongly dependent on the oxidation of dopamine, which could be tuned by either pH or by adding n-dodecanethiol. Interestingly, the thiol addition not only allowed the immobilization to take place in a wide pH range, but also led to superhydrophobicity of the resulting foams. Application of the superhydrophobic foams was illustrated by fast and selective collecting oils from water surface. Because catecholic derivatives exhibit high affinity to a variety of substances, the present strategy might be extendable to fabricate hybrid nanomaterials desirable for self-cleaning, environmental protection, sensors and catalysts, and so forth.


Measurement of free PSA in the diagnosis and staging of prostate cancer.
The purpose of this study was to analyze the role of total prostate-specific-antigen (PSA) and free-PSA levels in the diagnosis and clinical staging of prostate cancer. We determined total-PSA serum concentration and free-PSA percentage in 352 patients, 234 with benign prostatic hyperplasia (BPH) and 118 with prostate cancer. Clinical stage of patients with prostate cancer was T1 N0 M0 in 17, T2 N0 M0 in 27, T3-4 N0 M0 in 34 and T1-4 N0-3 M0-1 in 40. Median total-PSA serum concentration was 3.1 ng/ml in patients with BPH and 26.9 ng/ml in patients with prostate cancer, p < 0.0001. Median free-PSA level was 16.7% in patients with BPH and 8.1% in patients with prostate cancer, p < 0.0001. A cutpoint of 4.0 ng/ml detected 96.6% of the prostate cancer, but the percent rate of negative biopsies was 42.1%. For a free-PSA level of 25% in patients with total PSA greater than 4.0 ng/ml, the sensitivity was 97.4%, and the decrease in negative biopsies was 21%. Median total-PSA serum concentration in patients with prostate cancer according to clinical stage was 8.9 ng/ml for T1 N0 M0, 12.9 ng/ml for T2 N0 M0, 29.9 ng/ml for T3-4 N0 M0 and 317 ng/ml for T1-4 N1-3 M0-1, p < 0.001. Median free-PSA levels were 10.1%, 8.1%, 8.4% and 6.1% respectively, p > 0.05. According to the Gleason score, median total-PSA serum concentration was 10.6 ng/ml in patients between 2 and 4, 22.3 ng/ml between 5 and 7 and 77.0 ng/ml between 8 and 10, p < 0.05. Free PSA levels were 7.7%, 8.7% and 6.6% respectively, p > 0.05. Determination of percentage of free PSA appears to be a helpful method for enhancing the specificity of total PSA. A cutpoint of 25% could detect more than 95% of prostate cancers and avoid 21% of negative biopsies in patients with total PSA above 4.0 ng/ml. However, this parameter does not provide additional information about the clinical staging of prostate cancer.


Chemical-induced inflammation and inflammatory diseases.
It is now known that human exposure to certain chemicals e.g. benzene, halocarbons, ketones, nitrosamines, etc. can result in adverse health effects that are often not easily recognised as manifestations of chemical toxicity. These are inflammatory states, such as hepatitis, nephritis, scleroderma, and lupus, due to production of reactive oxygen species (ROS) through activation of cytochrome P4502E1 by the chemical, or by metabolism of the chemical to reactive intermediates and neoantigens which initiate immunotoxic effects. Intracellular glutathione (GSH), vitamins C, E and A protect against this ROS toxicity and inflammation; fasting and consumption of alcohol exacerbate it. Chronic inflammatory states may subsequently develop, including rheumatoid disease, atherosclerosis, diabetes, infertility and birth defects, multiple system organ failure (MSOF), Alzheimer's disease, and cancer.


The Karyote physico-chemical genomic, proteomic, metabolic cell modeling system.
Modeling approaches to the dynamics of a living cell are presented that are strongly based on its underlying physical and chemical processes and its hierarchical spatio-temporal organization. Through the inclusion of a broad spectrum of processes and a rigorous analysis of the multiple scale nature of cellular dynamics, we are attempting to advance cell modeling and its applications. The presentation focuses on our cell modeling system, which integrates data archiving and quantitative physico-chemical modeling and information theory to provide a seamless approach to the modeling/data analysis endeavor. Thereby the rapidly growing mess of genomic, proteomic, metabolic, and cell physiological data can be automatically used to develop and calibrate a predictive cell model. The discussion focuses on the Karyote cell modeling system and an introduction to the CellX and VirusX models. The Karyote software system integrates three elements: (1) a model-building and data archiving module that allows one to define a cell type to be modeled through its reaction network, structure, and transport processes as well as to choose the surrounding medium and other parameters of the phenomenon to be modeled; (2) a genomic, proteomic, metabolic cell simulator that solves the equations of metabolic reaction, transcription/translation polymerization and the exchange of molecules between parts of the cell and with the surrounding medium; and (3) an information theory module (ITM) that automates model calibration and development, and integrates a variety of data types with the cell dynamic computations. In Karyote, reactions may be fast (equilibrated) or slow (finite rate), and the special effects of enzymes and other minority species yielding steady-state cycles of arbitrary complexities are accounted for. These features of the dynamics are handled via rigorous multiple scale analysis. A user interface allows for an automated generation and solution of the equations of multiple timescale, compartmented dynamics. Karyote is based on a fixed intracellular structure. However, cell response to changes in the host medium, damage, development or transformation to abnormality can involve dramatic changes in intracellular structure. As this changes the nature of the cellular dynamics, a new model, CellX, is being developed based on the spatial distribution of concentration and other variables. This allows CellX to capture the self-organizing character of cellular behavior. The self-assembly of organelles, viruses, and other subcellular bodies is being addressed in a second new model, VirusX, that integrates molecular mechanics and continuum theory. VirusX is designed to study the influence of a host medium on viral self-assembly, structural stability, infection of a single cell, and transmission of disease.


[Which tools should be used to identify educational objectives in the regional health service? The experience in Marche Region (Italy)].
Through a specific methological path, educational objectives of Regional interest have been identified with a punctual analysis of Health Planning documents. While organizing each educational acrivities, all SSR departments indicate the Regional educational objectives they are linked with, each connected to national educational objectives. Continuing Medical Education program in the Marche Region is strictly connected with the acts of health planning from 2006. Each SSR department report accompanying the balance sheets from 2006 to 2009 must contain a section specifically dedicated to ECM Program, indicating the annual budget for ECM Program. All training activities are systematically monitored through a single information tool, shared by all SSR Departments and linked to the SSR Health Operators registry office. Strenghts of the ECM Educational Program of the Marche Region can be summarized as follows: guidance of Educational Program linked in a structured way to the Health Planning goals; punctual indications of redaction modes for PFA and Educational Reports; analytical scanning of activities/costs of the whole Regional Health Service training system, allowing Executive monitorings and reports, i.e. data that General Management can (... should) use to redirect not only training but their Company management as well.


Mouse extracellular superoxide dismutase: primary structure, tissue-specific gene expression, chromosomal localization, and lung in situ hybridization.
Extracellular superoxide dismutase (EC-SOD) is the major extracellular antioxidant enzyme. We have determined the primary structure of mouse EC-SOD by characterization of complementary DNA (cDNA) clones and by amino-acid sequence analysis of purified protein. cDNA sequence analysis indicates that mouse EC-SOD is synthesized as a 251-amino-acid precursor protein with a predicted molecular weight of 27,400 D. Amino-terminal micro sequence analysis of purified mature mouse lung EC-SOD demonstrated the sequence to begin with SSFDLADRLDPV-. These results indicate that EC-SOD as initially synthesized contains a 24-amino-acid precursor peptide, and that the mature protein is 227 amino acids in length. Computer algorithms that predict the most likely site of cotranslational signal peptidase cleavage suggest that processing will occur between amino acids 18 and 19 or 20 and 21, which implies that EC-SOD may be initially synthesized as a pre-pro-protein. Like human EC-SOD, mature mouse EC-SOD is glycosylated. The full-length mouse EC-SOD cDNA is 1,834 base pairs long and is 82% (79% for protein) identical to rat EC-SOD, but only 60% (60% for protein) identical to human EC-SOD. The mouse EC-SOD gene locus (Sod3) was mapped by interspecific backcross haplotype analysis as being 0.9 +/- 0.9 centimorgans distal to the Qdpr locus on mouse Chromosome 5, a position suggesting that the human homologue of EC-SOD will map close to the human QDPR locus (4p15.3). Of nine tissues examined by Northern blot analysis, those of the kidney and lung are by far the major tissues that express EC-SOD messenger RNA. Using in situ hybridization in the mouse lung, we demonstrate EC-SOD gene expression to be highly localized to alveolar Type II epithelial cells. These data suggest that alveolar Type II cells play a central role in mediating EC-SOD antioxidant function in the lung.


Peripheral blood CD34+ cells efficiently engraft human cytokine knock-in mice.
Human CD34+ hematopoietic stem and progenitor cells (HSPCs) can reconstitute a human hemato-lymphoid system when transplanted into immunocompromised mice. Although fetal liver-derived and cord blood-derived CD34+ cells lead to high engraftment levels, engraftment of mobilized, adult donor-derived CD34+ cells has remained poor. We generated so-called MSTRG and MISTRG humanized mice on a Rag2-/-Il2rg-/- background carrying a transgene for human signal regulatory protein α (SIRPα) and human homologs of the cytokine macrophage colony-stimulating factor, thrombopoietin, with or without interleukin-3 and granulocyte-macrophage colony-stimulating factor under murine promoters. Here we transplanted mobilized peripheral blood (PB) CD34+ cells in sublethally irradiated newborn and adult recipients. Human hematopoietic engraftment levels were significantly higher in bone marrow (BM), spleen, and PB in newborn transplanted MSTRG/MISTRG as compared with nonobese diabetic/severe combined immunodeficient Il2rg-/- or human SIRPα-transgenic Rag2-/-Il2rg-/- recipients. Furthermore, newborn transplanted MSTRG/MISTRG mice supported higher engraftment levels of human phenotypically defined HSPCs in BM, T cells in the thymus, and myeloid cells in nonhematopoietic organs such as liver, lung, colon, and skin, approximating the levels in the human system. Similar results were obtained in adult recipient mice. Thus, human cytokine knock-in mice might open new avenues for personalized studies of human pathophysiology of the hematopoietic and immune system in vivo.


Whole Mitochondrial Genome Analysis in Turkish Patients with Mitochondrial Diseases
Mitochondrial diseases are a clinically heterogeneous group of rare hereditary disorders that are defined by a genetic defect predominantly affecting mitochondrial oxidative phosphorylation. Mitochondrial diseases are caused by mutations of genes encoded by either nuclear DNA or mitochondrial DNA. Hundreds of different mitochondrial DNA point mutations and large-scale mitochondrial DNA rearrangements have been shown to cause mitochondrial diseases including Kearns–Sayre syndrome, Leber's hereditary optic neuropathy, Leigh syndrome, myoclonic epilepsy with ragged-red fibers, mitochondrial encephalopathy lactic acidosis stroke. To investigate new variants that could be associated with mitochondrial diseases and to determine the effect of mitochondrial DNA mutations on the clinical spectrum. Cross-sectional study. We screened whole mitochondrial DNA genome using next-generation sequencing in 16 patients who are considered to have mitochondrial disease. CentoGene and Mikrogen Genetic Diseases Diagnostic Center's database were used to investigate sequence variants. Detected variants were evaluated in bioinformatic databases to determine pathogenicity and were classified as class 1 (pathogenic), class 2 (likely pathogenic), and class 3 (variant of uncertain significance) according to CentoGene-ACMG database. As a result of the study, 2 patients were diagnosed with Leigh syndrome as previously reported class 1 mutations in MT-ATP6 and MT-ND5 genes. Four variants were identified for the first time in literature and 2 variants, previously reported but with uncertain pathogenic effect, are thought to be associated with mitochondrial disease. Mitochondrial DNA screening should be among the primary clinical tests in patients with suspected mitochondrial disease to rule out DNA-associated mutations.


Information routing driven by background chatter in a signaling network.
Living systems are capable of processing multiple sources of information simultaneously. This is true even at the cellular level, where not only coexisting signals stimulate the cell, but also the presence of fluctuating conditions is significant. When information is received by a cell signaling network via one specific input, the existence of other stimuli can provide a background activity -or chatter- that may affect signal transmission through the network and, therefore, the response of the cell. Here we study the modulation of information processing by chatter in the signaling network of a human cell, specifically, in a Boolean model of the signal transduction network of a fibroblast. We observe that the level of external chatter shapes the response of the system to information carrying signals in a nontrivial manner, modulates the activity levels of the network outputs, and effectively determines the paths of information flow. Our results show that the interactions and node dynamics, far from being random, confer versatility to the signaling network and allow transitions between different information-processing scenarios.


Current exposure to persistent polychlorinated biphenyls (PCBs) and dichlorodiphenyldichloroethylene (p,p'-DDE) of Belgian students from food and dust.
Human exposure to individual polychlorinated biphenyl (PCB) congeners and dichloro-diphenyldichloroethylene (p,p'-DDE) through food (duplicate diets) and indoor dust ingestion was assessed for 19 Belgian students. The serum concentrations of the persistent PCB congeners in serum (PCB 118, 138, 153, 170, and 180) have been correlated with the individual intake through food and dust. Dietary intakes of SigmaPCBs ranged between 40 and 204 ng/day (median 133). PCB exposure through dust ingestion ranged between 0.1 and 0.8 ng/day (median 0.3) or 0.3 and 1.7 ng/day (median 0.8), assuming average dust ingestion (20 mg/day) and high dust ingestion rates (50 mg/day), respectively. Dietary intake of p,p'-DDE was comparable to that of PCBs with a range from 21 to 214 ng/day (median 92). The exposure to p,p'-DDE via dust ingestion ranged between 0.02 and 0.43 ng/day (median 0.17) or 0.05 and 1.09 ng/day (median 0.43), assuming average and high dust ingestion rates, respectively. Concentrations measured in blood serum were 28-153 ng/g lipid weight (lw) (median 74) and 32-264 ng/g lw (median 45) for SigmaPCBs and p,p'-DDE, respectively. Serum concentrations in the studied population are slightly lower compared to other European populations. In spite of the uncertainty associated with the dust ingestion rates, food was the predominant exposure pathway for each PCB congener and for p,p'-DDE in the studied population. Food intake contributed more than 99% of the combined PCB intake from food and dust. No significant positive correlations (p > 0.05) were observed between the serum concentrations of PCBs and p,p'-DDE and the total intake through food and dust for each participant. Instead, it is hypothesized that past and episodic higher current intakes are more important determinants of body burden than continuous background exposures at low levels.


Dynamics and Context-Dependent Roles of DNA Methylation.
DNA methylation is one of the most extensively studied epigenetic marks. It is involved in transcriptional gene silencing and plays important roles during mammalian development. Its perturbation is often associated with human diseases. In mammalian genomes, DNA methylation is a prevalent modification that decorates the majority of cytosines. It is found at the promoters and enhancers of inactive genes, at repetitive elements, and within transcribed gene bodies. Its presence at promoters is dynamically linked to gene activity, suggesting that it could directly influence gene expression patterns and cellular identity. The genome-wide distribution and dynamic behaviour of this mark have been studied in great detail in a variety of tissues and cell lines, including early embryonic development and in embryonic stem cells. In combination with functional studies, these genome-wide maps of DNA methylation revealed interesting features of this mark and provided important insights into its dynamic nature and potential functional role in genome regulation. In this review, we discuss how these recent observations, in combination with insights obtained from biochemical and functional genetics studies, have expanded our current knowledge about the regulation and context-dependent roles of DNA methylation in mammalian genomes.