The acute pulmonary and thrombotic effects of cerium oxide nanoparticles after intratracheal instillation in mice.
Cerium oxide nanoparticles (CeO2 NPs), used as a diesel fuel catalyst, can be emitted into the ambient air, resulting in exposure to humans by inhalation. Recent studies have reported the development of lung toxicity after pulmonary exposure to CeO2 NPs. However, little is known about the possible thrombotic effects of these NPs. The present study investigated the acute (24 hours) effect of intratracheal (IT) instillation of either CeO2 NPs (0.1 or 0.5 mg/kg) or saline (control) on pulmonary and systemic inflammation and oxidative stress and thrombosis in mice. CeO2 NPs induced a significant increase of neutrophils into the bronchoalveolar lavage (BAL) fluid with an elevation of tumor necrosis factor α (TNFα) and a decrease in the activity of the antioxidant catalase. Lung sections of mice exposed to CeO2 NPs showed a dose-dependent infiltration of inflammatory cells consisting of macrophages and neutrophils. Similarly, the plasma levels of C-reactive protein and TNFα were significantly increased, whereas the activities of catalase and total antioxidant were significantly decreased. Interestingly, CeO2 NPs significantly and dose dependently induced a shortening of the thrombotic occlusion time in pial arterioles and venules. Moreover, the plasma concentrations of fibrinogen and plasminogen activator inhibitor-1 were significantly elevated by CeO2 NPs. The direct addition of CeO2 NPs (1, 5, or 25 μg/mL) to mouse whole blood, collected from the inferior vena cava, in vitro neither caused significant platelet aggregation nor affected prothrombin time or partial thromboplastin time, suggesting that the thrombotic events observed in vivo may have resulted from systemic inflammation and/or oxidative stress induced by CeO2 NPs. This study concludes that acute pulmonary exposure to CeO2 NPs induces pulmonary and systemic inflammation and oxidative stress and promotes thrombosis in vivo.


The leukocyte beta 1 integrins.
We summarize publications appearing in the past year on the blood cell beta 1 integrins VLA-1 through -6, including characterization of their ligand interactions, activation epitopes, signaling mechanisms, cellular distribution, and in vivo relevance. These studies extend our understanding of the beta 1 integrins, and continue to underscore their importance in leukocyte biology, which derives from their central role in mechanisms of cellular adhesion, growth, and differentiation.


Habitual coffee consumption and 24-h blood pressure control in older adults with hypertension.
In normotensive and hypertensive individuals, blood pressure (BP) rises acutely during a few hours following coffee or caffeine consumption. However, the effect of habitual coffee consumption on BP and BP control is uncertain. The aim of this study was to assess the association of habitual coffee consumption on 24-h BP and BP control among older adults with hypertension. Data were taken from the Seniors-Study on Nutrition and Cardiovascular Risk in Spain (ENRICA), a cross-sectional study conducted in 2012 among 1164 individuals aged ≥63 years. Habitual coffee consumption was assessed with a validated diet history. BP was recorded by 24-h ambulatory monitoring. Ambulatory hypertension was defined as BP ≥ 130/80 mm Hg or being under antihypertensive treatment, and uncontrolled BP was deemed as BP ≥ 130/80 mm Hg among hypertensives. Analyses were performed with linear and logistic regressions adjusted for the main confounders, including diet, time since diagnosis of hypertension and the number of antihypertensive drugs. Among the 715 hypertensive participants, those consuming ≥3 cups of coffee/day showed higher 24-h systolic BP (beta: 3.25 mm Hg, p value = 0.04) and diastolic BP (beta: 2.24 mm Hg, p value = 0.02) than non-coffee drinkers. Compared to non-coffee drinkers, the odds ratios (95% confidence interval) for uncontrolled BP among those consuming 1, 2, and ≥3 cups of coffee/day were, respectively: 1.95 (1.15-3.30), 1.41 (0.75-2.68), and 2.55 (1.28-5.09); p for trend = 0.05. The association was similar among individuals who were smokers, had excess weight (body mass index ≥25 kg/m2), low adherence to the Mediterranean diet, or hypercholesterolemia. No association was found between coffee consumption and having a non-dipper BP pattern (<10% nocturnal decline in BP) among hypertensives. Habitual coffee consumption was associated with uncontrolled BP in a hypertensive older population.


Cosmetic gynecology in the view of evidence-based medicine and ACOG recommendations: a review.
To conduct a methodological review of the existing scientific literature within the field of cosmetic gynecology in the view of evidence-based medicine and to establish their relevance to the ACOG Committee Opinion No. 378. The appropriate medical subject heading terms were selected and applied in the search of the Internet multiple databases since 1900 until January 2010. Articles focusing on cosmetic gynecology were reviewed. Also, anecdotal and advertising literatures were analyzed. A methodological review of the literatures was conducted. In peer review journals, 72 relevant articles related to cosmetic gynecology were identified. Anecdotal information was identified in 3 sources and over 1,100 published marketing literatures were identified on the Internet and no scientific journals. Among reviewed articles on cosmetic gynecology, only two articles met the level II-2 in evidence-based medicine. The absence of documentations on the safety and effectiveness of cosmetic vaginal procedures in the scientific literatures was ACOG's main concern. Practicing cosmetic gynecology within ACOG recommendations is desirable and possible. Currently, the standard of practice of cosmetic gynecology cannot be determined due to the absence of the documentation on safety and effectiveness. Traditional gynecologic surgical procedures cannot be called cosmetic procedures, since it is a deceptive form of practice and marketing. Creating medical terminology trademarks and establishing a business model that tries to control clinical-scientific knowledge dissemination is unethical.


Improvement of the ReaxFF Description for Functionalized Hydrocarbon/Water Weak Interactions in the Condensed Phase.
The ReaxFF protein reactive force field (protein-2013) has been successfully employed to simulate the biomolecules and membrane fuel cells, but it inaccurately describes the weak interaction of functionalized hydrocarbon/water molecules in condensed phase, especially for the density. In this article, the development of a ReaxFF force field (CHON-2017_weak) on the basis of protein-2013 is presented that improves the weak interaction description for atom pairs of carbon, hydrogen, oxygen, and nitrogen. To examine the quality of the force field, we performed a series of molecular dynamics simulations with model systems. These simulations, describing density trends for pure and mixture compound systems, demonstrate that CHON-2017_weak force field predictions are in good agreement with experimental data. Furthermore, ReaxFF can also describe the phase separation in hexane-water mixture and dissolution of ethanol or tetramethylammonium (TMA) in liquid water. To validate it in the application of membrane fuel cells, we studied structural property and degradation mechanism of TMA in alkaline aqueous solution, as well as some typical chemical reactions for small compounds. On the basis of our results, an additional reaction pathway is proposed for the degradation of TMA, which seems to be more energetically favorable compared to the main mechanism predicted from quantum mechanics calculations.


Effect of relative humidity on the airborne survival of rotavirus SA11.
Rotavirus SA11, suspended in tryptose phosphate broth with 2.5 mg of rhodamine B per ml, was aerosolized (Collison nebulizer) into a rotating drum, and the aerosols were held at 20 +/- 1 degree C with the desired relative humidity (RH). An all-glass impinger with tryptose phosphate broth was used to collect 1-min (5.6-liter) samples of air from the drum. The virus was found to survive best at medium (50 +/- 5%) RH, where its half-life was nearly 40 h. The half-life of the virus at the low (25 +/- 5%) RH level was about 9 h. Even at 72 h of aerosol age, 45 and 21% of the infectious virus remained detectable in the air at the medium and low RH levels, respectively. The high (80 +/- 5%) RH level was found to be the least favorable to the survival of the virus, since 50% of the infectious virus became undetectable within 2 h of aerosolization. In a separate experiment at the midrange RH, 3% of the infectious virus was detectable in the drum air after 223 h (9 days) of aerosol age. Rotaviruses could, therefore, survive in air for prolonged periods, thus making air a possible vehicle for their dissemination.


Protein delivery into live cells by incubation with an endosomolytic agent.
We report that a tetramethylrhodamine-labeled dimer of the cell-penetrating peptide TAT, dfTAT, penetrates live cells by escaping from endosomes with high efficiency. By mediating endosomal leakage, dfTAT also delivers proteins into cultured cells after a simple co-incubation procedure. We achieved cytosolic delivery in several cell lines and primary cells and observed that only a relatively small amount of material remained trapped inside endosomes. Delivery did not require a binding interaction between dfTAT and a protein, multiple molecules could be delivered simultaneously, and delivery could be repeated. dfTAT-mediated delivery did not noticeably affect cell viability, cell proliferation or gene expression. dfTAT-based intracellular delivery should be useful for cell-based assays, cellular imaging applications and the ex vivo manipulation of cells.


Designing Hydrogels for 3D Cell Culture Using Dynamic Covalent Crosslinking.
Designing simple biomaterials to replicate the biochemical and mechanical properties of tissues is an ongoing challenge in tissue engineering. For several decades, new biomaterials have been engineered using cytocompatible chemical reactions and spontaneous ligations via click chemistries to generate scaffolds and water swollen polymer networks, known as hydrogels, with tunable properties. However, most of these materials are static in nature, providing only macroscopic tunability of the scaffold mechanics, and do not reflect the dynamic environment of natural extracellular microenvironment. For more complex applications such as organoids or co-culture systems, there remain opportunities to investigate cells that locally remodel and change the physicochemical properties within the matrices. In this review, advanced biomaterials where dynamic covalent chemistry is used to produce stable 3D cell culture models and high-resolution constructs for both in vitro and in vivo applications, are discussed. The implications of dynamic covalent chemistry on viscoelastic properties of in vitro models are summarized, case studies in 3D cell culture are critically analyzed, and opportunities to further improve the performance of biomaterials for 3D tissue engineering are discussed.


Identification of dimethyl sulfoxide reductase in Actinobacillus pleuropneumoniae and its role in infection.
Actinobacillus pleuropneumoniae, the causative agent of porcine pleuropneumonia, is capable of persisting in oxygen-deprived surroundings, namely, tonsils and sequestered necrotic lung tissue. Utilization of alternative terminal electron acceptors in the absence of oxygen is a common strategy in bacteria under anaerobic growth conditions. In an experiment aimed at identification of genes expressed in vivo, the putative catalytic subunit DmsA of anaerobic dimethyl sulfoxide reductase was identified in an A. pleuropneumoniae serotype 7 strain. The 90-kDa protein exhibits 85% identity to the putative DmsA protein of Haemophilus influenzae, and its expression was found to be upregulated under anaerobic conditions. Analysis of the unfinished A. pleuropneumoniae genome sequence revealed putative open reading frames (ORFs) encoding DmsB and DmsC proteins situated downstream of the dmsA ORF. In order to investigate the role of the A. pleuropneumoniae DmsA protein in virulence, an isogenic deletion mutant, A. pleuropneumoniae DeltadmsA, was constructed and examined in an aerosol infection model. A. pleuropneumoniae DeltadmsA was attenuated in acute disease, which suggests that genes involved in oxidative metabolism under anaerobic conditions might contribute significantly to A. pleuropneumoniae virulence.


[When to start dialysis. The predialysis patient].
The incidence and prevalence of chronic renal failure (CKD), defined according to the NFK-KDOQI guidelines as a glomerular filtration rate less than 60 mL/min/1.73 m2 or the presence of microalbuminuria, is increasing worldwide, leading to an increased risk of cardiovascular disease. There is general agreement about the importance of early referral to the nephrologist and predialysis educational programs because this strategy prevents the progression (by the use of renin-angiotensin system blockers, low-protein diet) and complications (arterial hypertension, anemia, malnutrition, osteodystrophy, acidosis) of renal disease. Predialysis education helps patients choose the renal replacement therapy modality (hemodialysis, peritoneal dialysis, transplantation) and improve their quality of life. Furthermore, adequate predialysis care allows the nephrologist to prepare the vascular access well in advance. In contrast to the wished-for practice of early referral, patients are often referred to the nephrologist when renal failure is already advanced. This is mainly due to the fact that non-nephrologists pay little attention to identifying patients at risk for renal failure or defining the degree of renal failure according to the KDOQI classification. In addition, serum creatinine alone provides no adequate estimate of renal function, and both the MDRD equation and the Cockcroft-Gault formula permit a more accurate estimation of the glomerular filtration rate (GFR). Using the MDRD equation, the KDOQI guidelines recommend referral when GFR is less than 30 mL/min/1.73 m2. Late nephrology referral is an independent risk factor for early death while on dialysis, as well as being associated with a more frequent use of temporary catheters, particularly in the elderly but also in patients receiving regular nephrology care. This underlines the importance of a multidisciplinary predialysis approach which may bring additional benefits - beyond referral to a nephrologists - including a reduced hospitalization rate and improved survival. The KDOQI guidelines recommend evaluating the benefits and risks of starting renal replacement therapy when patients reach stage 5 (estimated GFR <15 mL/min/1.73 m2), although the ideal time for replacement therapy initiation remains a matter of debate and the results of prospective clinical trials are awaited to resolve this issue.


Monoclonal antibodies to gonococcal outer membrane protein I: location of a conserved epitope on protein IB.
Hybrid cell lines have been derived which produce monoclonal antibodies reacting with outer membrane protein I from Neisseria gonorrhoeae strain P9. The antibodies obtained showed variable reactivity with other strains but one antibody recognized an epitope present on all of the strains tested which expressed the protease sensitive protein IB. Purified IgG labelled with 125I was used in competitive radioimmunoassays with unlabelled antibody to investigate the spacial distribution of the epitopes recognized. Each pair of antibodies showed some degree of inhibition. The relative magnitude of inhibition suggested that the conserved epitope lies within a variable region containing other epitopes which determine the antigenic specificity of the protein. Western blotting of peptides derived by proteolytic digestion of protein IB revealed that the conserved epitope is located close to the chymotrypsin cleavage site within a 7000 Mr surface exposed region of the molecule.


Parallel on-axis holographic phase microscopy of biological cells and unicellular microorganism dynamics.
We apply a wide-field quantitative phase microscopy technique based on parallel two-step phase-shifting on-axis interferometry to visualize live biological cells and microorganism dynamics. The parallel on-axis holographic approach is more efficient with camera spatial bandwidth consumption compared to previous off-axis approaches and thus can capture finer sample spatial details, given a limited spatial bandwidth of a specific digital camera. Additionally, due to the parallel acquisition mechanism, the approach is suitable for visualizing rapid dynamic processes, permitting an interferometric acquisition rate equal to the camera frame rate. The method is demonstrated experimentally through phase microscopy of neurons and unicellular microorganisms.


Peri-transplant renal dysfunction in patients with non-alcoholic steatohepatitis undergoing liver transplantation.
Non-alcoholic fatty liver disease (NAFLD) is currently the most common etiology of chronic liver disease (CLD) caused by an accumulation of fat in the liver and globally is the leading indication of liver transplantation. Emerging data has recognized an increased association of NAFLD with risk of other metabolic liver diseases like type 2 diabetes mellitus, chronic kidney disease (CKD) and cardiovascular diseases. Pathophysiologically, NAFLD patients have a state of low-grade systemic inflammation, insulin resistance and atherogenic dyslipidemia which causes renal dysfunction. Patients with NAFLD cirrhosis awaiting liver transplant (LT) face unique challenges and have a significantly higher requirement of simultaneous-liver-kidney transplant as compared to other etiologies. Further, NAFLD not only recurs but also occurs as a de novo manifestation post-LT. There is also a significantly higher risk of waiting list stagnation and dropouts due to burdensome cardiometabolic disorders in NAFLD patients. The current review aims to understand the prevalence and pathogenetic basis of renal dysfunction in NAFLD. Additionally, the review describes the choice of immunosuppression protocols and use of intraoperative renal replacement therapy in context of intra and post-operative renal dysfunction in NAFLD patients. Prospective controlled trials focusing on NAFLD and development of CKD are needed to assess the existence of a causal and/or a bidirectional relationship between NAFLD and CKD.


Complete amino-acid sequence of the natural ATPase inhibitor from the mitochondria of the yeast Candida utilis.
The complete alignment of the 63 residues of the mitochondrial ATPase inhibitor from the yeast Candida utilis has been determined. The sequence study was carried out mainly by automatic (liquid and solid-phase) methods. Peptides were obtained by enzymatic digestion with clostripain and purified by reverse-phase high-performance liquid chromatography. The ATPase inhibitor contains three sets of repetitive sequences and eight clusters of charged residues, as also found in the inhibitor of Saccharomyces cerevisiae, with which it shares 58.7% homology of conserved residues. When the two yeast ATPase inhibitor sequences were compared to that of beef heart, 20 residues remained common to the three alignments, although the latter protein contained a long histidine-rich insertion, only found in this inhibitor. Most of the homologous residues were clustered near the center of the protein, which by partial proteolytic digestion of the beef heart ATPase inhibitor [Dianoux, A.C. et al. (1982) FEBS Lett. 140, 223-228] has already been shown to be involved in the biological function.


Prevalence of Chronic Illness and Health Seeking Behaviour in Malaysian Population: Results from the Third National Health Morbidity Survey (NHMS III) 2006.
The third National Health Morbidity Survey was conducted in 2006 on a nationally representative sample of population in Malaysia in order to obtain community-based data and information on the prevalence of chronic illness. Of 57,500 eligible respondents 56710 (98.6%) participated in the study. Estimated overall prevalence of chronic illness in the Malaysian population within a recall period of one year was 15.5% (95% CI 15.1% - 15.9%). Chronic illness was reported significantly higher among the females, 16.8% (16.3 - 17.3). The most common chronic illness was hypertension (7.9%, 7.6 - 8.2), followed by diabetes mellitus (4.0%, 3.8 - 4.2) and highest reported by the Indians (19.7%, 18.4 - 21.0). Among the respondents who had sought treatment for chronic illness from government health facilities, Malays (65.8%) and those with monthly household income of less than RM400 (76.6%) were the highest. Chinese (44.5%) and those with household income of RM5000 and above (54.3%) were the highest groups who sought treatment from the private health facilities. Most of the respondents reported mild illness was the main reason for not seeking treatment for their chronic illness. It is hoped that the results of this survey will help the Ministry of Health Malaysia to enhance health programmes and planning resource allocation in order to improve health status of the population.


Colorimetric optical nanosensors for trace explosive detection using metal nanoparticles: advances, pitfalls, and future perspective.
Warfare threats and acts of terror are challenging situations encountered by defense agencies across the globe and are of growing concern to the general public, and security-minded policy makers. Detecting ultra-low quantities of explosive compounds in remote locations or under harsh conditions for anti-terror purposes as well as the environmental monitoring of residual or discarded explosives in soil, remains a major challenge. The use of metal nanoparticles (NPs) for trace explosive detection has drawn considerable interest in recent years. For nano-based explosive sensor devices to meet real-life operational demands, analytical parameters such as, long-shelf life, stability under harsh conditions, ease-of-use, high sensitivity, excellent selectivity, and rapid signal response must be met. Generally, the analytical performance of colorimetric-based nanosensor systems is strongly dependent on the surface properties of the nanomaterial used in the colorimetric assay. The size and shape properties of metal NPs, surface functionalisation efficiency, and assay fabrication methods, are factors that influence the efficacy of colorimetric explosive nanosensor systems. This review reports on the design and analytical performances of colorimetric explosive sensor systems using metal NPs as optical signal transducers. The challenges of trace explosive detection, advances in metal NP colorimetric explosive design, limitations of each methods, and possible strategies to mitigate the problems are discussed.


Inhibition by yeast-derived mannoproteins of adherence to and invasion of Caco-2 cells by Campylobacter jejuni.
The main objective of the present work was to study the influence of yeast-derived mannoproteins on the adherence to and invasion of Caco-2 cells by Campylobacter jejuni. Mannoprotein fractions were prepared by enzymatic and thermal extraction methods. The method used to prepare the mannoprotein extracts influenced their composition and determined the efficacy of the extract against C. jejuni adherence and/or invasion. The availability of mannose in the mannoprotein fraction seemed to be important for inhibiting effective adherence and invasion of Caco-2-cells by C. jejuni, although protein moieties also played a role in the process. The study of the mechanisms involved in the inhibition of C. jejuni adherence and invasion by mannoproteins may have further implications in the control of this foodborne pathogen.


[Assessment of the Gleason score in biopsies and specimens of radical prostatectomy].
To evaluate the similarity between Gleason grade on prostate biopsies and their final result after radical prostatectomy. We retrospectively review the medical records of 129 patients who underwent radical prostatectomy. Mean PSA value was 10.7 ng/ml. The relationships between pathological reports of biopsies and radical prostatectomy specimens, and other variables such as PSA were established. Globally, 72 cases (55.8%) were in the same risk group by grade on biopsy and surgical specimen. Biopsy result was understaged in 48 cases (37.2%) and overstaged in 9 cases (7%). We found biopsy understaging in 42 cases (60%) in the low grade group, 6 cases (10.9%) in the intermediate grade group, and 50% in the high grade group, although in this latter the number of cases was very low. Overall biopsy understaging was 37.2%, being much more evident in low grade tumors (60%). It seems that understaging was greater when PSA > 10 ng/ml, although differences were not significant.


Gestalt factors modulate basic spatial vision.
Human perception of a stimulus varies depending on the context in which the stimulus is presented. Such contextual modulation has often been explained by two basic neural mechanisms: lateral inhibition and spatial pooling. In the present study, we presented observers with a vernier stimulus flanked by single lines; observers' ability to discriminate the offset direction of the vernier stimulus deteriorated in accordance with both explanations. However, when the flanking lines were part of a geometric shape (i.e., a good Gestalt), this deterioration strongly diminished. These findings cannot be explained by lateral inhibition or spatial pooling. It seems that Gestalt factors play an important role in contextual modulation. We propose that contextual modulation can be used as a quantitative measure to investigate the rules governing the grouping of elements into meaningful wholes.


A novel method for incorporating respiratory-matched attenuation correction in the motion correction of cardiac PET-CT studies.
Mismatches between PET and CT datasets due to respiratory effects can lead to artefactual perfusion defects. To overcome this, we have proposed a method of aligning a single CT with each frame of a gated PET study in a semi-automatic manner, incorporating a statistical shape model of the diaphragm and a rigid registration of the heart. This ensures that the structures that could influence the appearance of the reconstructed cardiac activity are correctly matched between emission and transmission datasets. When tested on two patient studies, it was found in both cases that attenuation correction using the proposed technique resulted in PET images that were closer to the gold standard of attenuation correction with a gated CT, compared with scenarios where only heart matching was considered (and not the diaphragm) or where no transformation was performed (i.e. where a single CT frame was used to attenuation-correct all PET frames). These preliminary results suggest that diaphragm matching between PET and CT improves the quantitative accuracy of reconstructed PET images and that the proposed method of using a statistical shape model to describe the diaphragm shape and motion, in combination with a rigid registration to determine respiratory-induced heart motion, is a feasible method of achieving this.


2-Methoxyestradiol ameliorates paraquat-induced pulmonary fibrosis by inhibiting the TGF-β1/Smad2/3 signaling pathway.
Paraquat (PQ) is a highly effective and highly toxic herbicide that is highly toxic to both humans and animals. Pulmonary fibrosis is the primary cause of fatality in patients with PQ poisoning, there is no effective drug treatment yet. 2-Methoxyestradiol (2ME) is a natural metabolite of estradiol with anti-tumor, anti-angiogenesis, and anti-proliferative effects. Whether 2ME has the potential to inhibit pulmonary fibrosis induced by PQ is unclear. This study aims to investigate the potential effects and mechanism of 2ME on PQ-induced pulmonary fibrosis. C57BL/6 mice and A549 cells were exposed to PQ to establish pulmonary fibrosis model. In vivo, Hematoxylin and eosin (H&E) staining was utilized to assess the pathological characteristics. Masson's trichrome staining was employed to evaluate the collagen deposition. Western blot and immunohistochemistry were conducted to determine the expressions of fibrosis markers. In vitro, the expressions of epithelial-mesenchymal transition (EMT) markers were detected using western blot and immunofluorescence to evaluated the potential inhibition of PQ-induced EMT by 2ME. And proteins associated with the TGF-β1/Smad2/3 signaling pathway were measured by western blot in vivo and in vitro. The result found that 2ME can ameliorated PQ-induced pulmonary fibrosis and inhibit the activation of TGF-β1/Smad2/3 signaling pathway. These findings suggest that 2ME may serve as a potential therapeutic agent for treating PQ-induced pulmonary fibrosis.


NLRX1 inhibits the early stages of CNS inflammation and prevents the onset of spontaneous autoimmunity.
Nucleotide-binding, leucine-rich repeat containing X1 (NLRX1) is a mitochondria-located innate immune sensor that inhibits major pro-inflammatory pathways such as type I interferon and nuclear factor-κB signaling. We generated a novel, spontaneous, and rapidly progressing mouse model of multiple sclerosis (MS) by crossing myelin-specific T-cell receptor (TCR) transgenic mice with Nlrx1-/- mice. About half of the resulting progeny developed spontaneous experimental autoimmune encephalomyelitis (spEAE), which was associated with severe demyelination and inflammation in the central nervous system (CNS). Using lymphocyte-deficient mice and a series of adoptive transfer experiments, we demonstrate that genetic susceptibility to EAE lies within the innate immune compartment. We show that NLRX1 inhibits the subclinical stages of microglial activation and prevents the generation of neurotoxic astrocytes that induce neuronal and oligodendrocyte death in vitro. Moreover, we discovered several mutations within NLRX1 that run in MS-affected families. In summary, our findings highlight the importance of NLRX1 in controlling the early stages of CNS inflammation and preventing the onset of spontaneous autoimmunity.


Traumatic brain injury in children--clinical implications.
Traumatic brain injury (TBI) is the leading cause of death in childhood; however only very few studies focusing on the specific pathophysiology and treatment have been published to date. Head trauma is more likely in young children than in adults given the same deceleration of the body due to their large and heavy heads and weak cervical ligaments and muscles. Resulting brain injury is more severe due to their thin, pliable skulls and the yet unfused sutures. Accordingly, children below the age of 4 years have lower chances of a full recovery after severe TBI, although in general, neurologic recovery after severe brain injury in children is better than in adults. The time course of brain injury can be divided into two steps: primary and secondary injury. Primary brain injury exclusively results from the initial impact. In contrast, adverse physiologic conditions during recovery after head trauma may account for additional brain damage, which is then referred to as secondary brain injury. As primary brain injury can only be influenced by preventive measures, all therapeutic efforts during the post-injury period focus on the reduction of secondary injury to the traumatized brain. Several mechanisms have been identified to be involved in the development of post-traumatic secondary brain injury, which render the rationale for the key treatment strategies. Three evidence based measures are of critical importance to prevent or minimize secondary brain injury: (1) avoid hypoxemia, (2) avoid post-traumatic arterial hypotension, and (3) refer the traumatized child to an experienced trauma team at a center that provides the availability of special equipment, e.g. for surgical procedures and airway management, for this age group. For several other therapeutical means, e.g. hypothermia or specific surgical interventions, clinical evidence to date is insufficient to allow recommendation as rescue treatment for children at risk of severe neurological sequelae following TBI. This review discusses the clinical implication of pathophysiologic mechanisms of TBI in the developing brain according to the recent literature and current guidelines. It follows the clinical approach to a head injured child, that can be divided into three phases, i.e. initial assessment and stabilization, followed by first tier, and if necessary second tier therapeutic interventions to assure adequate oxygenation and perfusion of the brain.


Three-dimensional microplate-enhanced alveolar ridge augmentation--an alternative to nerve transposition.
The placement of endosseous implants in the atrophic posterior mandible has presented a challenge to the surgeon and restorative dentist. Traditional procedures either require nerve transposition or utilization of implants placed at less than favorable angles. A technique is presented for the three-dimensional augmentation of the atrophic alveolar ridge utilizing microplate-enhanced guided bone regeneration technology. The applications of this technique in other intraoral and extraoral sites are currently under investigation by the authors.


A multifaceted health-service intervention in remote Aboriginal communities: 3-year follow-up of the impact on diabetes care.
To examine the trends in processes of diabetes care and in participant outcomes after an intervention in two remote regions of Australia. Follow-up study over 3 years. Seven health centres in the Tiwi Islands and the Katherine West region of the Northern Territory. 137 Aboriginal people with type 2 diabetes. Implementation of a multifaceted trial, including transfer of purchasing and planning responsibility to local health boards, the development and dissemination of clinical guidelines supported by electronic registers, recall and reminder systems and associated staff training, and audit and feedback. Trends in the proportion of Aboriginal people receiving services in accordance with clinical guidelines and in the proportion for whom specified levels of blood pressure and glycosylated haemoglobin (HbA(1c)) were achieved; health staff perceptions of barriers to effective service delivery. An initial improvement in overall service levels from 40% to 49% was not fully sustained over the 3-year period. The overall proportion of services delivered varied from 22% to 64% between communities and over time. The proportion of participants whose most recent HbA(1c) level was less than 7% improved from 19% to 32%, but there was little change in blood pressure control. Perceived barriers to service delivery included discontinuities in staffing, lack of work-practice support and patients' acceptance of services. Multifaceted interventions can improve quality of care in this environment, but achieving sustainable, high-quality care in a range of services and local conditions presents particular challenges. Developing and testing strategies for consistent and sustained improvement should be a priority for service providers and researchers.


Nutritional interventions in sarcopenia: where do we stand?
To document recent evidence regarding the role of nutrition as an intervention for sarcopenia. A review of seven randomized controlled trials (RCTs) on beta-hydroxy-beta-methylbutyrate (HMB) alone on muscle loss in 147 adults showed greater muscle mass gain in the intervention group, but no benefit in muscle strength and physical performance measures. Three other review articles examined nutrition and exercise as combined intervention, and suggest enhancement of benefits of exercise by nutrition supplements (energy, protein, vitamin D). Four trials reported on nutrition alone as intervention, mainly consisting of whey protein, leucine, HMB and vitamin D, with variable results on muscle mass and function. Four trials examined the combined effects of nutrition combined with exercise, showing improvements in muscle mass and function. To date, evidence suggests that nutrition intervention alone does have benefit, and certainly enhances the impact of exercise. Nutrients include high-quality protein, leucine, HMB and vitamin D. Long-lasting impact may depend on baseline nutritional status, baseline severity of sarcopenia, and long-lasting adherence to the intervention regime. Future large-scale multicentered RCTs using standardized protocols may provide evidence for formulating guidelines on nutritional intervention for sarcopenia. There is a paucity of data for nursing home populations.


Decreased expression of N-myc precedes retinoic acid-induced morphological differentiation of human neuroblastoma.
Proto-oncogenes may be important in the cellular processes central for the growth and differentiation of normal cells. N-myc is a DNA sequence which shares limited homology to the proto-oncogene c-myc and has been found to be amplified in both primary tissue and cell lines from neuroblastoma, a childhood tumour of neuroectodermal origin. Differentiation of this embryonal tumour is of clinical importance, since occasional tumours have been noted to differentiate in vivo to benign ganglioneuroma. In vitro, many human neuroblastoma cell lines can be induced to differentiate morphologically and biochemically by a variety of agents. Retinoic acid (RA), an analogue of vitamin A, has been shown to inhibit neuroblastoma cell growth and clonability in soft agar, and to induce extensive neurite outgrowth. Therefore we examined the relationship of N-myc expression to the in vitro differentiation of these cells. We report here that in the case of RA-induced differentiation, a decreased level of expression is detected within 6 h of treatment and precedes both cell-cycle changes and morphological differentiation.


Prenatal iodine deficiency results in structurally and functionally immature lungs in neonatal rats.
Maternal hypothyroidism affects postnatal lung structure. High prevalence of hypothyroxinemia (low T4, normal T3) in iodine-deficient pregnant women and associated risk for neuropsychological development along with high infant/neonatal mortality ascribed to respiratory distress prompted us to study the effects of maternal hypothyroxinemia on postnatal lung development. Female Sprague Dawley rats were given a low-iodine diet (LID) with 1% KClO(4) in drinking water for 10 days, to minimize thyroid hormone differences. Half of these rats were continued on iodine-deficient diet; ID (LID with 0.005% KClO(4)) for 3 mo, whereas the rest were switched to an iodine-sufficient diet; IS [LID + potassium iodide (10 μg iodine/20 g of diet + normal drinking water)]. Pups born to ID mothers were compared with age-matched pups from IS mothers at postnatal days 8 (P8) and 16 (P16) (n = 6-8/group). ID pups had normal circulating T3 but significantly low T4 levels (P < 0.05) and concomitantly approximately sixfold higher thyroid hormone receptor-β mRNA in alveolar epithelium. Lung histology revealed larger and irregularly shaped alveoli in ID pups relative to controls. Lung function was assessed at P16 using a double-chambered plethysmograph and observed reduced tidal volume, peak inspiratory and expiratory flow, and dynamic lung compliance in ID pups compared with IS pups. Significant lowering of surfactant protein (SP)-B and SP-C mRNA and protein found in ID pups at P16. ID pups had 16-fold lower matrix metalloproteinase-9 mRNA levels in their alveolar epithelium. In addition, mRNA levels of thyroid transcription factor-1 and SP-D were significantly higher (3-fold) compared with IS pups. At P16, significantly lower levels of SP-B and SP-C found in ID pups may be responsible for immature lung development and reduced lung compliance. Our data suggest that maternal hypothyroxinemia may result in the development of immature lungs that, through respiratory distress, could contribute to the observed high infant mortality in ID neonates.


Correlations of Inflammatory Factors with Intestinal Flora and Gastrointestinal Incommensurate Symptoms in Children with Asthma.
BACKGROUND Bronchial asthma is a common pediatric disease, the pathogenesis of which is complicated. The correlations of the levels of inflammatory factors in peripheral serum with intestinal flora and gastrointestinal incommensurate symptoms in children with asthma remain to be further elucidated. MATERIAL AND METHODS A total of 70 children diagnosed with asthma in the Pediatric Department of our hospital from February 2016 to March 2017 were enrolled as an observation group, and another 25 healthy children in the same age range were selected as a control group. The levels of inflammatory factors [C-reactive protein (CRP), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6)], the total load of intestinal flora, and the main strains were detected among all included patients. Moreover, incommensurate symptoms of patients in the observation group were recorded and gastrointestinal symptom rating scale (GSRS) scores were calculated. The differences in indexes between the observation group and the control group were compared. RESULTS The levels of CRP, TNF-α, and IL-6 in peripheral serum in the observation group were significantly higher than those in the control group (p<0.05). The analysis of the correlations of inflammatory factors in peripheral serum with intestinal flora and GSRS scores showed that C-reactive protein (CRP) was positively correlated with GSRS scores (r=0.696, p<0.001) and the total load of intestinal bacteria (r=0.813, p<0.001). CONCLUSIONS The inflammatory factors in peripheral serum of children with asthma are closely correlated with intestinal flora and gastrointestinal function. With the increasingly high levels of inflammatory factors in peripheral serum, the probability of intestinal flora disturbance and gastrointestinal incommensurate symptoms will be increased.


Efficacy of water spray for evaporative cooling in athletes with spinal cord injury.
Interventional crossover study. Spinal cord injury (SCI) disrupts afferent input to the hypothalamus and impairs efferent vaso- and sudomotor output, especially in lesions above the sympathetic chain (T1-L2). In consequence, persons with SCI under heat stress experience impairment in the ability to dissipate heat proportional to the lesion level. Thermoregulatory dysfunction places an individual at high risk of hyperthermia, which can be life threatening, especially for athletes with SCI during exercise. Current evidence on therapeutic cooling techniques in athletes with SCI is limited, but basic physiologic and research data suggest water spray (WS) might be efficacious, particularly in athletes with tetraplegia (TP), who are most impaired in thermoregulation. The aim of this study was to evaluate the effect of WS on core temperature (Tc) during exercise in athletes with SCI. Texas, USA. Eleven individuals with SCI: seven with TP, four with paraplegia (PP); and sixteen able-bodied (AB) controls underwent a wheelchair intermittent sprint exercise for 90 min under two conditions: (1) WS application every 15 min and (2) control (C), without WS. Tc was measured every 15 min and was analyzed for the effect of group (TP, PP, and AB) and time. Change in Tc (ΔTc) was also compared between groups. ΔTc was significantly higher in TP vs. PP (p < 0.0001) and TP vs. AB (p < 0.0001) groups under C treatment. WS significantly attenuated ΔTc in TP (p = 0.001), but did not change ΔTc in PP or AB. WS effectively attenuated Tc elevation during exercise in athletes with TP. Texas chapter of the Paralyzed Veterans of America.


Notch Signaling in T Helper Cell Subsets: Instructor or Unbiased Amplifier?
For protection against pathogens, it is essential that naïve CD4+ T cells differentiate into specific effector T helper (Th) cell subsets following activation by antigen presented by dendritic cells (DCs). Next to T cell receptor and cytokine signals, membrane-bound Notch ligands have an important role in orchestrating Th cell differentiation. Several studies provided evidence that DC activation is accompanied by surface expression of Notch ligands. Intriguingly, DCs that express the delta-like or Jagged Notch ligands gain the capacity to instruct Th1 or Th2 cell polarization, respectively. However, in contrast to this model it has also been hypothesized that Notch signaling acts as a general amplifier of Th cell responses rather than an instructive director of specific T cell fates. In this alternative model, Notch enhances proliferation, cytokine production, and anti-apoptotic signals or promotes co-stimulatory signals in T cells. An instructive role for Notch ligand expressing DCs in the induction of Th cell differentiation is further challenged by evidence for the involvement of Notch signaling in differentiation of Th9, Th17, regulatory T cells, and follicular Th cells. In this review, we will discuss the two opposing models, referred to as the "instructive" and the "unbiased amplifier" model. We highlight both the function of different Notch receptors on CD4+ T cells and the impact of Notch ligands on antigen-presenting cells.


Triple therapy and initial renal graft perfusion.
Cyclosporin A may decrease the perfusion of transplanted kidneys, especially if they are ischaemic. Therefore, monitoring of graft perfusion was performed with intravenous 99mTc pertechnetate angiography in 25 patients within 36 hours after renal transplantation and repeated 24 or 48 hours later and then again 48 hours later. In all patients, immunosuppression was performed with triple therapy (low dose prednisone, azathioprine, and cyclosporin A) which included a delay in cyclosporin A administration to the second postoperative day. Treatment with triple therapy did not cause a decrease in renal blood flow in the days immediately after transplantation as single therapy with high-dose cyclosporin A has been reported to do.


The effects of prohibiting gestures on children's lexical retrieval ability.
Two alternative accounts have been proposed to explain the role of gestures in thinking and speaking. The Information Packaging Hypothesis (Kita, 2000) claims that gestures are important for the conceptual packaging of information before it is coded into a linguistic form for speech. The Lexical Retrieval Hypothesis (Rauscher, Krauss & Chen, 1996) sees gestures as functioning more at the level of speech production in helping the speaker to find the right words. The latter hypothesis has not been fully explored with children. In this study children were given a naming task under conditions that allowed and restricted gestures. Children named more words correctly and resolved more 'tip-of-the-tongue' states when allowed to gesture than when not, suggesting that gestures facilitate access to the lexicon in children and are important for speech production as well as conceptualization.


[Autonomy, privacy and the implementation of the principle of "informed consent" with regard to nursing intervention from the view point of the aged].
The aim of this study was to describe patient autonomy, privacy and the implementation of the principle of informed consent in the care of elderly patients in facilities experienced by themselves. This study is part of the BIOMED 2 project "Patients' autonomy and privacy in nursing interventions" supported by the European Commission. Interview data (n = 95) were collected among elderly people in German facilities for geriatrics and in nursing homes. The results showed there was a lack of opportunity by the elderly people to make self-determined decisions. The principle of "informed consent" was hardly realised. The participants felt their privacy was not respected in multi-bedded rooms and in situations of dressing and eliminating. One can proceed on the assumption that the lack of information, the need of help and the fixed organizing structures of the facilities are the reasons why elderly people play a rather passive role as patients. It might be possible to improve the autonomy of elderly people if the nurses as an advocate supported them to make self-determined decisions. The implementation of the principle of informed consent with regard to nursing interventions would promote both autonomy and respect of privacy. Furthermore, one can assume that the autonomy and quality of life of elderly people could be promoted if the organizing structures of the facilities were more flexible.


Detection of thiol modifications by hydrogen sulfide.
Hydrogen sulfide (H2S) is an important gasotransmitter in both animals and plants. Many physiological events, including responses to stress, have been suggested to involve H2S, at least in part. On the other hand, numerous responses have been reported following treatment with H2S, including changes in the levels of antioxidants and the activities of transcription factors. Therefore, it is important to understand and unravel the events that are taking place downstream of H2S in signaling pathways. H2S is known to interact with other reactive signaling molecules such as reactive oxygen species (ROS) and nitric oxide (NO). One of the mechanisms by which ROS and NO have effects in a cell is the modification of thiol groups on proteins, by oxidation or S-nitrosylation, respectively. Recently, it has been reported that H2S can also modify thiols. Here we report a method for the determination of thiol modifications on proteins following the treatment with biological samples with H2S donors. Here, the nematode Caenorhabditis elegans is used as a model system but this method can be used for samples from other animals or plants.


Vonoprazan-associated Gastric Mucosal Redness: A Report of Four Cases.
To date, no cases of vonoprazan-associated gastric mucosal redness have been reported, and its endoscopic and pathological features remain largely unclear. We report four cases of vonoprazan-associated gastric mucosal redness. In all cases, esophagogastroduodenoscopy (EGD) demonstrated linear or spotty redness that newly appeared in the greater curvature of the middle gastric body after the initiation of vonoprazan, which disappeared after its discontinuation. A tissue biopsy taken from the gastric mucosa with redness revealed inflammatory cell infiltration, parietal cell protrusions (PCPs), and oxyntic gland dilatation. To our knowledge, this is the first report describing the endoscopic and pathological features of vonoprazan-associated gastric mucosal redness.


Age-related changes in cardiac structure and function in Fischer 344 x Brown Norway hybrid rats.
The effects of aging on cardiovascular function and cardiac structure were determined in a rat model recommended for gerontological studies. A cross-sectional analysis assessed cardiac changes in male Fischer 344 x Brown Norway F1 hybrid rats (FBN) from adulthood to the very aged (n = 6 per 12-, 18-, 21-, 24-, 27-, 30-, 33-, 36-, and 39-mo-old group). Rats underwent echocardiographic and hemodynamic analyses to determine standard values for left ventricular (LV) mass, LV wall thickness, LV chamber diameter, heart rate, LV fractional shortening, mitral inflow velocity, LV relaxation time, and aortic/LV pressures. Histological analyses were used to assess LV fibrotic infiltration and cardiomyocyte volume density over time. Aged rats had an increased LV mass-to-body weight ratio and deteriorated systolic function. LV systolic pressure declined with age. Histological analysis demonstrated a gradual increase in fibrosis and a decrease in cardiomyocyte volume density with age. We conclude that, although significant physiological and morphological changes occurred in heart function and structure between 12 and 39 mo of age, these changes did not likely contribute to mortality. We report reference values for cardiac function and structure in adult FBN male rats through very old age at 3-mo intervals.


Electrolyte and hormonal effects of deoxycorticosterone acetate in young pigs.
Balances of sodium, potassium, and water were studied in the growing male pig as hypertension developed in response to subcutaneous implantation of deoxycorticosterone acetate (DOCA). Serum sodium, potassium, deoxycorticosterone (DOC), aldosterone, and plasma renin activity (PRA) were determined. These variables were observed in a total of 10 experimental and nine control pigs. All animals were uninephrectomized and fed a diet of Purina Pig Chow and tap water ad libitum. No salt was added to the food or water. Serum DOC levels rose dramatically on the day of the implantation, then gradually declined but remained approximately 10 times greater than control levels 40 days after implant. Plasma renin activity was suppressed rapidly and completely, whereas aldosterone fell only slowly to about half its control value. Sodium retention was maximum during the first 24 hours. Therefore an "escape" process became operative, causing sodium balance to return to normal after the third day, at which time the major rise in arterial pressure occurred. A marked increase in water turnover (intake and output) also began after the third day and persisted throughout the experimental period. Water balance remained normal during this period of increased turnover. Hypokalemia developed in the absence of kaliuresis, suggesting that potassium moved into the cells. Except for the potassium retention, these changes parallel the abnormalities seen in other states of mineralocorticoid excess.


A direct aspiration first-pass technique vs stentriever thrombectomy in emergent large vessel intracranial occlusions.
OBJECTIVE Endovascular thrombectomy in patients with acute ischemic stroke caused by occlusion of the proximal anterior circulation arteries is superior to standard medical therapy. Stentriever thrombectomy with or without aspiration assistance was the predominant technique used in the 5 randomized controlled trials that demonstrated the superiority of endovascular thrombectomy. Other studies have highlighted the efficacy of a direct aspiration first-pass technique (ADAPT). METHODS To compare the angiographic and clinical outcomes of ADAPT versus stentriever thrombectomy in patients with emergent large vessel occlusions (ELVO) of the anterior intracranial circulation, the records of 134 patients who were treated between June 2012 and October 2015 were reviewed. RESULTS Within this cohort, 117 patients were eligible for evaluation. ADAPT was used in 47 patients, 20 (42.5%) of whom required rescue stentriever thrombectomy, and primary stentriever thrombectomy was performed in 70 patients. Patients in the ADAPT group were slightly younger than those in the stentriever group (63.5 vs 69.4 years; p = 0.04); however, there were no differences in the other baseline clinical or radiographic factors. Procedural time (54.0 vs 77.1 minutes; p < 0.01) and time to a Thrombolysis in Cerebral Infarction (TICI) scale score of 2b/3 recanalization (294.3 vs 346.7 minutes; p < 0.01) were significantly lower in patients undergoing ADAPT versus stentriever thrombectomy. The rates of TICI 2b/3 recanalization were similar between the ADAPT and stentriever groups (82.9% vs 71.4%; p = 0.19). There were no differences in the rates of symptomatic intracranial hemorrhage or procedural complications. The rates of good functional outcome (modified Rankin Scale Score 0-2) at 90 days were similar between the ADAPT and stentriever groups (48.9% vs 41.4%; p = 0.45), even when accounting for the subset of patients in the ADAPT group who required rescue stentriever thrombectomy. CONCLUSIONS The present study demonstrates that ADAPT and primary stentriever thrombectomy for acute ischemic stroke due to ELVO are equivalent with respect to the rates of TICI 2b/3 recanalization and 90-day mRS scores. Given the reduced procedural time and time to TICI 2b/3 recanalization with similar functional outcomes, an initial attempt at recanalization with ADAPT may be warranted prior to stentriever thrombectomy.


The role of the pediatrician in the management of children with genital ambiguities.
To present the diagnostic criteria of genital ambiguity, the initial medical management and the attitude expected of pediatricians. Review of the scientific literature in the form of articles indexed on MEDLINE, in English and Portuguese, published between 1990 and 2007 and dealing with the pediatric age group. Pediatricians have a fundamental role to play in the assessment of genital ambiguity, the purpose of which is to arrive at an etiologic diagnosis in the shortest possible time in order to define the patient's sex and plan treatment. There are specific diagnostic criteria, but, in general, genitalia are ambiguous whenever there is difficulty in attributing gender to a child. The pediatrician should inform the patient's family that assignment of their child's sex will depend upon detailed laboratory investigations, preferably carried out by a multidisciplinary team at a tertiary service. The 46,XX or 46,XY karyotypes are not alone sufficient to define the gender of rearing, although the test is fundamental to guide the investigation. When there are no palpable gonads, the first hypothesis should be congenital adrenal hyperplasia. Other causes included partial androgen insensitivity, 5alpha-reductase deficiency, partial gonadal dysgenesis and hermaphroditism. The family should be provided with support and information throughout the assessment process, and their participation is fundamental in the decision of which gender to rear the child in. Although cases of genital ambiguity are relatively rare for pediatricians, they should be well-informed on the subject and the correct management of these conditions, since they will often be responsible for the initial guidance that families receive and for maintaining contact between them and the multidisciplinary team.


Modulation of Respiratory TLR3-Anti-Viral Response by Probiotic Microorganisms: Lessons Learned from Lactobacillus rhamnosus CRL1505.
Respiratory syncytial virus (RSV) is the leading cause of lower respiratory tract illness in infants and young children. Host immune response is implicated in both protective and immunopathological mechanisms during RSV infection. Activation of Toll-like receptor (TLR)-3 in innate immune cells by RSV can induce airway inflammation, protective immune response, and pulmonary immunopathology. A clear understanding of RSV-host interaction is important for the development of novel and effective therapeutic strategies. Several studies have centered on whether probiotic microorganisms with the capacity to stimulate the immune system (immunobiotics) might sufficiently stimulate the common mucosal immune system to improve defenses in the respiratory tract. In this regard, it was demonstrated that some orally administered immunobiotics do have the ability to stimulate respiratory immunity and increase resistance to viral infections. Moreover, during the last decade scientists have significantly advanced in the knowledge of the cellular and molecular mechanisms involved in the protective effect of immunobiotics in the respiratory tract. This review examines the most recent advances dealing with the use of immunobiotic bacteria to improve resistance against viral respiratory infections. More specifically, the article discuss the mechanisms involved in the capacity of the immunobiotic strain Lactobacillus rhamnosus CRL1505 to modulate the TLR3-mediated immune response in the respiratory tract and to increase the resistance to RSV infection. In addition, we review the role of interferon (IFN)-γ and interleukin (IL)-10 in the immunoregulatory effect of the CRL1505 strain that has been successfully used for reducing incidence and morbidity of viral airways infections in children.


Photoluminescent thermometry based on europium-activated calcium sulphide.
A photoluminescent thermometer, based on the transient emission response of a europium-activated calcium sulphide phosphor, is described in detail. This optical thermometry system has special advantages for temperature measurements in microwave and RF fields and potential application in electromagnetically induced clinical hyperthermia. A laboratory system has been constructed which utilises a fibreoptic probe with external diameter 0.8 mm. A system temperature resolution of 0.1 degree C has been achieved in the range 37-47 degrees C.


Breast cancer screening disparities among immigrant women by world region of origin: a population-based study in Ontario, Canada.
Rates of mammography screening for breast cancer are disproportionately low in certain subgroups including low-income and immigrant women. The purpose of the study was to examine differences in rates of appropriate breast cancer screening (i.e., screening mammography every 2 years) among Ontario immigrant women by world region of origin and explore the association between appropriate breast cancer screening among these women groups and individual and structural factors. A cohort of 183,332 screening-eligible immigrant women living in Ontario between 2010 and 2012 was created from linked databases and classified into eight world regions of origin. Appropriate screening rates were calculated for each region by age group and selected sociodemographic, immigration, and healthcare-related characteristics. The association between appropriate screening across the eight regions of origin and selected sociodemographic, immigration, and health-related characteristics was explored using multivariate Poisson regression. Screening varied by region of origin, with South Asian women (48.5%) having the lowest and Caribbean and Latin American women (63.7%) the highest cancer screening rates. Factors significantly associated with lower screening across the world regions of origin included living in the lowest income neighborhoods, having a refugee status, being a new immigrant, not having a regular physical examination, not being enrolled in a primary care patient enrollment model, having a male physician, and having an internationally trained physician. Multiple interventions entailing cross-sector collaboration, promotion of patient enrollment models, community engagement, comprehensive and intensive outreach to women, and knowledge translation and transfer to physicians should be considered to address screening disparities among immigrant population. Consideration should be given to design and delivery of culturally appropriate and easily accessible cancer screening programs targeted at high- risk immigrant subgroups, such as women of South Asian origin, refugees, and new immigrants.


First Case of Typhoid Fever due to Extensively Drug-resistant Salmonella enterica serovar Typhi in Italy.
Typhoid fever is a potentially severe and occasionally life-threatening bacteraemic illness caused by Salmonella enterica serovar Typhi (S. Typhi). In Pakistan, an outbreak of extensively drug-resistant (XDR) S. Typhi cases began in November 2016. We report on a five-year-old boy who contracted enteric fever while travelling in Pakistan and was diagnosed after returning to Italy in September 2019. Blood culture isolated Salmonella enterica serovar Typhi that was XDR to all first-line antibiotics, including ceftriaxone and fluoroquinolones. Empiric therapy was switched to meropenem, and the patient recovered completely. Whole-genome sequencing showed that this isolate was of haplotype H58. The XDR S. Typhi clone encoded a chromosomally located resistance region and harbored a plasmid encoding additional resistance elements, including the blaCTX-M-15 extended-spectrum β-lactamase and the qnrS fluoroquinolone resistance gene. This is the first case of typhoid fever due to XDR S. Typhi detected in Italy and one of the first paediatric cases reported outside Pakistan, highlighting the need to be vigilant for future cases. While new vaccines against typhoid are in development, clinicians should consider adapting their empiric approach for patients returning from regions at risk of XDR S. Typhi outbreak with typhoid symptoms.


Biologic and serologic characterization of the 161A strain of Naegleria fowleri.
The 161A strain of Naegleria isolated from the nasal swab of a boy (9) was grown axenically in Nelson's medium. When 10,000 amoebae from the axenic medium were inoculated onto each monkey kidney cell (Vero) culture, characteristic cytopathic effects (CPE) were noticed in 4 to 5 days. The CPE consisted of granulation of the host cell cytoplasm, cell shrinkage, nuclear pycnosis, and discontinuity of cell sheet. When 10,000 amoebae were instilled intranasally into a group of ten 2- to 3-week-old mice, 8 of the 10 mice exhibited characteristic symptoms of primary amoebic meningoencephalitis and died within 10 to 12 days. Histopathology of the brain revealed necrotic tissue and an acute inflammatory reaction in the superficial regions of the brain. In the gel diffusion and immunoelectrophoresis tests the sonically disrupted antigens of 161A amoebae reacted extensively with the hyper-immune sera against 3 strains (CA, CJ, HB-1) of pathogenic N. fowleri and produced patterns very similar to those produced by the homologous systems. Further, anti-HB-1 serum absorbed with the 161A antigens failed to react with the antigens of HB-1, CA, CJ, and 161A strains thus indicating antigenic identity of 161A strain with N. fowleri. In view of these observations it was concluded that the strain 161A is pathogenic and should be reclassified as N. fowleri.


Methods to determine effects of cranberry proanthocyanidins on extraintestinal infections: Relevance for urinary tract health.
Urinary tract infections (UTI) are one of the most frequent extraintestinal infections caused by Escherichia coli (ExPEC). Cranberry juice has been used for decades to alleviate symptoms and prevent recurrent UTI. The putative compounds in cranberries are proanthocyanidins (PAC), specifically PAC with "A-type" bonds. Since PAC are not absorbed, their health benefits in UTI may occur through interactions at the mucosal surface in the gastrointestinal tract. Recent research showed that higher agglutination of ExPEC and reduced bacterial invasion are correlated with higher number of "A-type" bonds and higher degree of polymerization of PAC. An understanding of PAC structure-activity relationship is becoming feasible due to advancements, not only in obtaining purified PAC fractions that allow accurate estimation, but also in high-resolution MS methodologies, specifically, MALDI-TOF MS. A recent MALDI-TOF MS deconvolution method allows quantification of the ratios of "A-type" to "B-type" bonds enabling characteristic fingerprints. Moreover, the generation of fluorescently labeled PAC allows visualization of the interaction between ExPEC and PAC with microscopy. These tools can be used to establish structure-activity relationships between PAC and UTI and give insight on the mechanism of action of these compounds in the gut without being absorbed.


Methyl mercury influences growth-related signaling in MCF-7 breast cancer cells.
Environmental contaminants have been shown to alter growth-regulating signaling pathways through molecular mechanisms that are mainly unclear. Here we report that within a narrow concentration range (0.5-1 microM) methyl mercury (MeHg) significantly stimulated growth of MCF-7 cells, induced Ca(2+) mobilization, and activated extracellular signal-regulated kinase (1/2) (Erk1/2). MeHg modulated E(2)-dependent stimulation of growth in a dose-dependent manner, although MeHg neither suppresses nor increases constitutive E(2) metabolism. MeHg demonstrated weak estrogen receptor (ER)-binding ability. However, long preincubation with antiestrogens LY(156,758) and ICI(164,384) decreased MeHg-induced foci formation, Ca(2+) mobilization, and Erk1/2 activation, confirming involvement of ERs. The MeHg-induced increase in [Ca(2+)](i) was observed to coincide with enhanced Erk1/2 phosphorylation. These data suggest that MeHg can significantly modulate the intracellular signaling environment in MCF-7 cells, resulting in a dose-dependent alteration of ER-mediated estrogenic capacity and therefore should be considered as a potential estrogen-disrupting compound.


Evaluation of Abbott CMV-M enzyme immunoassay for detection of cytomegalovirus immunoglobulin M antibody.
The Abbott CMV-M enzyme immunoassay (EIA) for the qualitative determination of immunoglobulin M (IgM) antibody to cytomegalovirus in human serum was compared with the indirect fluorescent-antibody (IFA) test on 338 human serum specimens. Discordant specimens were evaluated by IFA following isolation of IgM fractions. Discordant specimens remaining after IFA testing were evaluated by an IgM-specific EIA (CYTOMEGELISA M; M.A. Bioproducts). After resolution of discordant specimens, the CMV-M EIA was 94.7% sensitive and 99.1% specific.


Bovine Lactoferrin to Prevent Neonatal Infections in Low-Birth-Weight Newborns in Pakistan: Protocol for a Three-Arm Double-Blind Randomized Controlled Trial.
Sepsis is a common and severe complication in premature neonates, particularly those born with low birth weights (<2500 g). Neonatal sepsis is steadily emerging as a leading cause of neonatal mortality in Pakistan. Lactoferrin is a natural product with broad-spectrum antimicrobial properties and glycoprotein that is actively involved in innate immune host responses. Clinical trials have revealed its protective effect on sepsis, but lactoferrin dosage, duration, and role in the prevention of sepsis are still uncertain. We aimed to establish the efficacy of bovine lactoferrin in the prevention of late-onset sepsis and to determine the optimal dose and method of administering bovine lactoferrin that may contribute to improvement in overall survival of low birth weight infants. We will implement the study in 2 phases at the Aga Khan University Hospital. The first phase, which we have completed, was formative research. This phase mainly focused on a qualitative exploration of perceptions about feeding and caring practices of low birth weight newborns and a trial of improved practices for the preparation and administration of bovine lactoferrin to newborns. The second phase is a 3-arm double-blind randomized controlled trial. In this phase, we randomly allocated 2 different daily oral prophylactic doses of bovine lactoferrin (150 mg or 300 mg) and placebo to 300 low-birth weight neonates starting within the first 72 hours of birth and continuing for the first 28 days of life. The study protocol was approved by the Ethics Review Committee of Aga Khan University on August 16, 2017. Data collection began in April 2018 and was completed in September 2020. Data analyses are yet to be completed. We expect the results to be published in peer-reviewed journals by autumn of 2021. This intervention, if effective, has the potential to be translated into a safe, affordable, and widely utilized treatment to prevent sepsis and, subsequently, may improve the survival outcomes of low birth weight neonates in Pakistan and other low- and middle-income countries. ClinicalTrials.gov NCT03431558; https://clinicaltrials.gov/ct2/show/NCT03431558. PRR1-10.2196/23994.


Evolution of endogenous retroviruses in the Suidae: evidence for different viral subpopulations in African and Eurasian host species.
Porcine endogenous retroviruses (PERVs) represent remnants of an exogenous form that have become integrated in the domestic pig (Sus scrofa) genome. Although they are usually inactive, the capacity of γ1 ERVs to infect human cells in vitro has raised concerns about xenotransplantation because the viruses could cross the species barrier to humans. Here we have analyzed the evolution of γ1 ERVs in ten species of Suidae (suids, pigs and hogs) from Eurasia and Africa using DNA sequences for their coding domains (gag, pro/pol and env genes). For comparison with γ1 PERVs, we have also analysed γ2 ERVs which in domestic pigs are known to be inactive and do not pose a risk to xenotransplantation. Phylogenetic analysis using Bayesian inference showed that γ1 and γ2 ERVs have distinctive evolutionary histories. Firstly, two different viral lineages of γ1 ERVs were found and a coevolutionary analysis demonstrated that they correspond broadly to their host phylogeny, one of Eurasian and another of African species, and show no evidence of horizontal transmission. γ2 ERVs, however, show a bush-like evolution, suggesting a rapid viral radiation from a single common ancestor with no correspondence between host and viral evolutionary trees. Furthermore, though γ1 ERV env genes do not possess frequent stop codons, γ2 env genes do. To understand whether γ1 suid ERVs may be still replicating, we have also evaluated their likely mechanism of proliferation by statistically testing internal to terminal branches using nonsynonymous versus synonymous substitution ratios. Our results suggest that γ1 ERVs are increasing in copy number by reinfection, which requires the translocation of the virus from one cell to another. Evidence of at least two viral subpopulations was observed in γ1 ERVs from Eurasian and African host species. These results should be taken into account in xenotransplantation since γ1 ERVs appear to be codiverging with their host and maintaining ongoing capacity to infect somatic and germ cells.


Effect of conjugated bilirubin on clinical outcomes in infective endocarditis.
Liver dysfunction is associated with adverse events in infective endocarditis (IE). However, few studies have explored the predictive value of conjugated bilirubin (CB) in IE. We aimed to investigate the nature of the link between CB and adverse prognosis in patients with IE. Consecutive patients with IE between January 2009 and July 2015 were enrolled. Multivariate analysis was performed to confirm whether CB was an independent risk factor for adverse outcomes. In all, 1010 patients were included and divided into two groups according to admission CB level (μmol/L): normal (≤ 7.0, n = 820) and elevated (> 7.0, n = 190) CB groups. In-hospital mortality (5.0% vs. 22.1%, p < 0.001) and major adverse cardiac events (16.8% vs. 36.3%, p < 0.001) were significantly higher in patients with increased CB. A possible J-shaped relationship was found between CB and in-hospital events. Further, CB had more predictive power than total bilirubin in predicting in-hospital death (AUC 0.715 vs. 0.674, p = 0.010). Elevated CB was an independent predictor of in-hospital death (adjusted OR = 2.62, 95%CI 1.40-4.91, p = 0.003). Moreover, CB (increment 1 μmol/L) was independently associated with higher long-term mortality. Kaplan-Meier curves indicated that patients with elevated CB were associated with higher cumulative rate of long-term death (log-rank = 21.47, p < 0.001). CB, a biomarker of liver function, was a relatively powerful predictor of in-hospital and long-term adverse prognosis of IE and could likely comprise a novel risk evaluation strategy.


Effectiveness of early coronary angioplasty and abciximab for failed thrombolysis (reteplase or alteplase) during acute myocardial infarction (results from the GUSTO-III trial). Global Use of Strategies To Open occluded coronary arteries.
We evaluated the effects of abciximab treatment during early angioplasty after clinically failed thrombolysis for acute myocardial infarction. In the Global Use of Strategies To Open occluded coronary arteries (GUSTO-III) trial of reteplase versus alteplase for acute infarction (n = 15,059), 392 patients underwent angioplasty a median of 3.5 hours after thrombolysis and had complete procedural data. We compared 30-day mortality and in-hospital outcomes between patients who received abciximab (n = 83) and those who did not (n = 309), and (among patients given abciximab) between those randomized to alteplase versus reteplase. Patients given abciximab had anterior infarction less often, but were more often in Killip classes III or IV. The 30-day mortality rate tended to be lower with abciximab (3.6% vs 9.7%, p = 0.076), more so after adjustment for baseline differences (p = 0.042). The composite of death, stroke, or reinfarction did not differ significantly with abciximab treatment (12% vs 14%, p = 0.7), but it occurred less often among abciximab-treated patients who had been randomized to reteplase (n = 55) versus alteplase (n = 28) (7% vs 21%, p = 0.08). Severe bleeding was increased among abciximab-treated patients (3.6% vs 1.0%, p = 0.08), despite less heparin use. No intracranial hemorrhages occurred with abciximab. The use of abciximab for early angioplasty after clinically failed thrombolysis resulted in trends toward lower 30-day mortality and increased bleeding.


Oxytocin inhibits food and fluid intake in rats.
Increasing evidence indirectly suggests a role for oxytocinergic neurons in the control of ingestive behaviors. The present study was aimed at directly investigating a possible effect of oxytocin on food and water intake in rats. Oxytocin, whether administered intracerebroventricularly (ICV) (1-10 micrograms/rat) or intraperitoneally (IP) (375-3,000 micrograms/kg) dose dependently inhibited food intake in freely feeding animals; in schedule-fed animals fasting for 21 h, oxytocin not only reduced food intake but also reduced the time spent eating and increased the latency to first meal. On the other hand, oxytocin antagonist d(CH2)5Tyr(Me)-[Orn8]-vasotocin, ICV injected at the dose of 10 micrograms/rat, increased food intake and time spent eating and reduced the latency to first meal; moreover, it completely prevented the effect of oxytocin. Water intake was studied both in freely drinking animals and in three different models of thirst (water deprivation, hypertonic saline administration, angiotensin II injection). In all cases, oxytocin dose dependently inhibited water intake, in a dose range of 0.1-10 micrograms/rat (ICV) or 93-750 micrograms/kg (IP). In the water deprivation model, ICV pretreatment with d(CH2)5Tyr(Me)-[Orn8]-vasotocin completely prevented the antidipsogenic effect of oxytocin. In conclusion, these data show that oxytocin directly inhibits food and water intake in rats, the effect being specifically mediated by brain oxytocin receptors. This may support the idea that the brain oxytocinergic system plays an important role in the regulation of ingestive behaviors.


Controversial opinions on the role of cocaine and amphetamine-regulated transcript (CART) in prolactin release.
Cocaine and amphetamine regulated transcript (CART) is widely expressed in the central nervous system and in several endocrine organs. The physiological role of this peptide includes modulation of appetite control, energy expenditure, thermoregulation and hormone secretion. It has been suggested that CART influences prolactin (PRL) secretion both directly and indirectly. However, the mechanism underlying the regulation of PRL release by CART remains unclear.


Identification of MT-45 Metabolites: In Silico Prediction, In Vitro Incubation with Rat Hepatocytes and In Vivo Confirmation.
MT-45 is a synthetic opioid with a pharmacological activity comparable to morphine and it has been involved in intoxications and fatalities reported in Europe and in USA. It was recently subject to control measures, but to date the metabolic pathways of the substance are still unknown. Using rat hepatocytes and LC-HRMS, 14 novel Phase I and II MT-45 metabolites were identified, products of monohydroxylation, dihydroxylation and N-dealkylation; glucuronide conjugation of mono- and dihydroxylated metabolites also occurred. The detected metabolites were firstly predicted in silico, then incubation of the drug with rat hepatocytes was carried out and the obtained metabolites were identified by LC-HRMS, with retention times, mass shift between theoretical mass and observed mass (<5 ppm), peak abundance and fragmentation pattern. Hydroxylated MT-45 was found to be the major metabolite of MT-45 in vitro experiments. The presence of all metabolites was confirmed by in vivo experiments in urine samples of CD-1 male mice; in these samples hydroxy-MT-45-glucuronide and di-hydroxy-MT-45-glucuronide are the most abundant metabolites, while the parent drug is found at concentration <10 ng mL-1 after 300 min. The knowledge of Phase I and II MT-45 metabolite structure is then crucial to develop analytical methods to identify MT-45 consumption in clinical and forensic testing.


An antimitotic and antivascular agent BPR0L075 overcomes multidrug resistance and induces mitotic catastrophe in paclitaxel-resistant ovarian cancer cells.
Paclitaxel plays a major role in the treatment of ovarian cancer; however, resistance to paclitaxel is frequently observed. Thus, new therapy that can overcome paclitaxel resistance will be of significant clinical importance. We evaluated antiproliferative effects of an antimitotic and antivascular agent BPR0L075 in paclitaxel-resistant ovarian cancer cells. BPR0L075 displays potent and broad-spectrum cytotoxicity at low nanomolar concentrations (IC50 = 2-7 nM) against both parental ovarian cancer cells (OVCAR-3, SKOV-3, and A2780-1A9) and paclitaxel-resistant sublines (OVCAR-3-TR, SKOV-3-TR, 1A9-PTX10), regardless of the expression levels of the multidrug resistance transporter P-gp and class III β-tubulin or mutation of β-tubulin. BPR0L075 blocks cell cycle at the G2/M phase in paclitaxel-resistant cells while equal concentration of paclitaxel treatment was ineffective. BPR0L075 induces cell death by a dual mechanism in parental and paclitaxel-resistant ovarian cancer cells. In the parental cells (OVCAR-3 and SKOV-3), BPR0L075 induced apoptosis, evidenced by poly(ADP-ribose) polymerase (PARP) cleavage and DNA ladder formation. BPR0L075 induced cell death in paclitaxel-resistant ovarian cancer cells (OVCAR-3-TR and SKOV-3-TR) is primarily due to mitotic catastrophe, evidenced by formation of giant, multinucleated cells and absence of PARP cleavage. Immunoblotting analysis shows that BPR0L075 treatment induced up-regulation of cyclin B1, BubR1, MPM-2, and survivin protein levels and Bcl-XL phosphorylation in parental cells; however, in resistant cells, the endogenous expressions of BubR1 and survivin were depleted, BPR0L075 treatment failed to induce MPM-2 expression and phosphorylation of Bcl-XL. BPR0L075 induced cell death in both parental and paclitaxel-resistant ovarian cancer cells proceed through caspase-3 independent mechanisms. In conclusion, BPR0L075 displays potent cytotoxic effects in ovarian cancer cells with a potential to overcome paclitaxel resistance by bypassing efflux transporters and inducing mitotic catastrophe. BPR0L075 represents a novel microtubule therapeutic to overcome multidrug resistance and trigger alternative cell death by mitotic catastrophe in ovarian cancer cells that are apoptosis-resistant.


Is Nonoperative Management the Best First-line Option for High-grade Renal trauma? A Systematic Review.
The management of high-grade (Grade IV-V) renal injuries remains controversial. There has been an increase in the use of (NOM) but limited data exists comparing outcomes with open surgical exploration. To conduct a systematic review to determine if NOM is the best first-line option for high-grade renal trauma in terms of safety and effectiveness. Medline, Embase, and Cochrane Library were searched for all relevant publications, without time or language limitations. The primary harm outcome was overall mortality and the primary benefit outcome was renal preservation rate. Secondary outcomes included length of hospital stay and complication rate. Single-arm studies were included as there were few comparative studies. Only studies with more than 50 patients were included. Data were narratively synthesised in light of methodological and clinical heterogeneity. The risk of bias of each included study was assessed. Seven nonrandomised comparative and four single-arm studies were selected for data extraction. Seven hundred and eighty-seven patients were included from the comparative studies with 535 patients in the NOM group and 252 in the open surgical exploration group. A further 825 patients were included from single-arm studies. Results from comparative studies: overall mortality: NOM (0-3%), open surgical exploration (0-29%); renal preservation rate: NOM (84-100%), open surgical exploration (0-82%); complication rate: NOM (5-32%), open surgical exploration (10-76%). Overall mortality and renal preservation rate were significantly better in the NOM group whereas there was no statistical difference with regard to complication rate. Length of hospital stay was found be significantly reduced in the NOM group. Patients in the open surgical exploration group were more likely to have Grade V injuries, have a lower systolic blood pressure, and higher injury severity score on admission. No randomised controlled trials were identified and significant heterogeneity existed with regard to outcome reporting. However, NOM appeared to be safe and effective in a stable patient with a higher renal preservation rate, a shorter length of stay, and a comparable complication rate to open surgical exploration. Overall mortality was higher in the open surgical exploration group, though this was likely due to selection bias. The data of this systematic review suggest nonoperative management continues to be favoured to surgical exploration in the management of high-grade renal trauma whenever possible. However, comparisons between both interventions are difficult as patients who have surgery are often more seriously injured than those managed nonoperatively, and existing studies do not report on outcomes consistently.


Synergistic interactions of physiologic increments of glucagon, epinephrine, and cortisol in the dog: a model for stress-induced hyperglycemia.
To evaluate the role of anti-insulin hormone actions and interactions in the pathogenesis of stress-induced hyperglycemia, the counterregulatory hormones, glucagon, epinephrine, and cortisol were infused alone as well as in double and triple combinations into normal conscious dogs in doses that were designed to simulate changes observed in severe stress. Infusion of glucagon, epinephrine, or cortisol alone produced only mild or insignificant elevations in plasma glucose concentration. In contrast, the rise in plasma glucose produced by combined infusion of any two counterregulatory hormones was 50-215% greater (P < 0.005-0.001) than the sum of the respective individual infusions. Furthermore, when all three hormones were infused simultaneously, the increment in plasma glucose concentration (144+/-2 mg/dl) was two- to fourfold greater than the sum of the responses to the individual hormone infusions or the sum of any combination of double plus single hormone infusion (P < 0.001). Infusion of glucagon or epinephrine alone resulted in a transient rise in glucose production (as measured by [3-(3)H]glucose). While glucagon infusion was accompanied by a rise in glucose clearance, with epinephrine there was a sustained, 20% fall in glucose clearance. When epinephrine was infused together with glucagon, the rise in glucose production was additive, albeit transient. However, the inhibitory effect of epinephrine on glucose clearance predominated, thereby accounting for the exaggerated glycemic response to combined infusion of glucagon and epinephrine. Although infusion of cortisol alone had no effect on glucose production, the addition of cortisol markedly accentuated hyperglycemia produced by glucagon and(or) epinephrine primarily by sustaining the increases in glucose production produced by these hormones. The combined hormonal infusions had no effect on beta-hydroxybutyrate concentration. It is concluded that (a) physiologic increments in glucagon, epinephrine, and cortisol interact synergistically in the normal dog so as to rapidly produce marked fasting hyperglycemia; (b) in this interaction, epinephrine enhances glucagon-stimulated glucose output and interferes with glucose uptake while cortisol sustains elevations in glucose production produced by epinephrine and glucagon; and (c) these data indicate that changes in glucose metabolism in circumstances in which several counterregulatory hormones are elevated (e.g., "stress hyperglycemia") are a consequence of synergistic interactions among these hormones.


GluR3 subunit regulates sleep, breathing and seizure generation.
The functional role of GluR3 AMPA (alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid) receptor subunits has remained elusive. In vitro studies of genetic knockout mice have not yielded significant alterations in synaptic communication. However, behavioural approaches utilizing knockout mice have shown that the subunit may be involved in exploration and motor coordination, suggesting that in vivo methodologies may be more forthcoming. We tested the hypothesis that GluR3 subunits are involved in the modulation of neural network activity. We used a freely behaving mouse model to examine the effect of GluR3(-/-) on field potential recordings of electroencephalogram, vital functions (i.e. breathing and heart rate) and muscle tone across natural sleep and wakefulness states. We found that GluR3(-/-) mice virtually lack electroencephalographic signatures of NREM sleep (n = 9) as demonstrated by reduction in electroencephalogram power in the low-frequency bands (delta1, delta2 and theta). In addition, three of nine GluR3(-/-) mice expressed seizure activity during wakefulness and sleep, suggesting that deletion of the GluR3 gene may predispose to seizure. GluR3 gene knockout also produced state-dependent respiratory modulation, with a selective reduction in breathing rate during behavioural inactivity. These findings show that GluR3 subunits have diverse neurophysiological impact, modulating oscillatory networks for sleep, breathing and seizure generation. Finally, this is the first study to demonstrate the feasibility of direct diaphragm electromyogram recordings in freely behaving mice.


Expression of low density lipoprotein receptor messenger ribonucleic acid in placentas from pregnancies with intrauterine growth retardation.
The low density lipoprotein receptor plays an important role in the uptake of maternal plasma lipoproteins for placental steroid metabolism. Using an RNase protection assay, the expression of low density lipoprotein receptor mRNA was measured in human placentas from pregnancies in the second and third trimester and from pregnancies with intrauterine growth retardation. Pregnancies complicated by intrauterine growth retardation showed a significantly higher expression (1.28), compared with the term and preterm pregnancies (P < 0.05). The increased low density lipoprotein receptor expression in placentas from pregnancies with intrauterine growth retardation could be a compensatory mechanism for the lower circulating low density lipoprotein concentrations in women with intrauterine growth retardation.


[Electroacupuncture protects the brain against acute ischemic injury via up-regulation of delta-opioid receptor in rats].
To explore the effect of delta-opioid receptor (DOR) in electroacupuncture (EA) protecting the brain against acute ischemic injury. Fifty-one rats were randomly divided into sham ischemia group, ischemia group, sham EA group, EA group, and EA+DOR antagonist (naltrindole) group. Transient focal cerebral ischemia (1 hour) was induced in rat brain by middle cerebral artery occlusion (MCAO) method. EA was applied on Shuigou (GV 26) and Neiguan (PC 6) for 30 min, starting immediately after the onset of reperfusion. Neurological deficit scores and volume of cerebral infarction were detected after 24-hour reperfusion. Other 12 rats were randomly divided into sham ischemia group, ischemia group, EA group and EA + naltrindole group. DOR protein expressions were assessed by Western blotting after 24-hour reperfusion. In comparison with the ischemia group and sham EA group, EA significantly reduced ischemic infarction and neurological deficits (P<0.05); EA significantly increased the expression of 60 kD DOR protein (P<0.05) and tended to increase that of 36 kD DOR protein (P>0.05). When naltrindole was combined with EA, the naltrindole completely abolished the EA-induced protection in ischemic infarction and neurological deficits, and also arrested the expression of DOR. EA can up-regulate DOR expression and protect the brain from ischemia-reperfusion injury.


Opioid use disorder treatment and the role of New Jersey Medicaid policy changes: perspectives of office-based buprenorphine providers.
Background: In the US, seventy percent of drug-related deaths are attributed to opioids. In response to the ongoing opioid crisis, New Jersey's (NJ) Medicaid program implemented the MATrx model to increase treatment access for Medicaid participants with opioid use disorder (OUD). The model's goals include increasing the number of office-based treatment providers, enhancing Medicaid reimbursement for certain treatment services, and elimination of prior authorizations for OUD medications.Objectives: To explore office-based addiction treatment providers' experiences delivering care in the context of statewide policy changes and their perspectives on treatment access changes and remaining barriers.Methods: This qualitative study used purposive sampling to recruit office-based New Jersey medications for opioid use disorder (MOUD) providers . Twenty-two providers (11 females, 11 males) discussed treatment experiences since the policy changes in 2019, including evaluations of the current state of OUD care in New Jersey and perceived outcomes of the MATrx model policy changes.Results: Providers reported the MOUD climate in NJ improved as Medicaid implemented policies intended to reduce barriers to care and increase treatment access. Elimination of prior authorizations was noted as important, as it reduced provider burden and allowed greater focus on care delivery. However, barriers remained, including stigma, pharmacy supply issues, and difficulty obtaining injectable or non-generic medication formulations.Conclusion: NJ policies may have improved access to care for Medicaid beneficiaries by reducing barriers to care and supporting providers in prescribing MOUD. Yet, stigma and lack of psychosocial supports still need to be addressed to further improve access and care quality.


Identification of N-benzothiazolyl-2-benzenesulfonamides as novel ABCA1 expression upregulators.
ATP binding cassette transporter A1 (ABCA1) is a critical transporter that mediates cellular cholesterol efflux from macrophages to apolipoprotein A-I (ApoA-I). Therefore, increasing the expression level of ABCA1 is anti-atherogenic and ABCA1 expression upregulators have become novel choices for atherosclerosis treatment. In this study, a series of N-benzothiazolyl-2-benzenesulfonamides, based on the structure of WY06 discovered in our laboratory, were designed and synthesized as novel ABCA1 expression upregulators. Based on an in vitro ABCA1 upregulatory cell model, ABCA1 upregulation of target compounds was evaluated. Compounds 6c, 6d, and 6i have good upregulated ABCA1 expression activities, with EC50 values of 0.97, 0.37, and 0.41 μM, respectively. A preliminary structure-activity relationship is summarized. Replacing the methoxy group on the benzothiazole moiety of WY06 with a fluorine or chlorine atom and exchanging the ester group with a cyano group resulted in more potent ABCA1 upregulating activity. Moreover, compound 6i increased ABCA1 mRNA and protein expression and significantly promoted cholesterol efflux in RAW264.7 cells. In conclusion, N-benzothiazolyl-2-benzenesulfonamides were identified as novel ABCA1 expression upregulators.


Exploitation of autochthonous Tuscan sourdough yeasts as potential starters.
The increasing demand for healthy baked goods boosted studies on sourdough microbiota with beneficial metabolic traits, to be used as potential functional starters. Here, yeast populations of traditional sourdoughs collected from four Tuscan bakeries were investigated. Among 200 isolated strains, 78 were randomly selected and molecularly characterized. Saccharomyces cerevisiae was dominant, representing the only species detected in three out of the four sourdoughs. The fourth one harbored also Kazachstania humilis. Inter-delta regions analysis revealed a high intraspecific polymorphism discriminating 16 biotypes of S. cerevisiae isolates, which clustered based on their origin. Representative isolates from each biotype group were individually used to ferment soft and durum wheat flour, aiming at evaluating their pro-technological, nutritional and functional features. During fermentation under standardized conditions, all strains were able to grow of ca. 2 log cycles, but only S. cerevisiae L10Y, D18Y and D20Y had a significantly shorter latency phase in both flours. Overall, the highest volumes were reached after 16 h of fermentation in both soft and durum fermented dough. S. cerevisiae D2Y produced the highest dough volume increase. K. humilis G23Y was the only strain able to increase the total free amino acids concentration of the doughs. Overall, values of phytase activity were significantly higher in durum compared to the corresponding soft fermented dough. K. humilis G23Y and S. cerevisiae D20Y, D24Y showed a threefold higher phytase activity than spontaneously fermented control, and the highest concentration of total phenols. Almost all the strains led to increases of antioxidant activity, without significant differences among them. Investigations on the resistance of the strains to simulated gastric and intestinal conditions, that is considered a pre-requisite for the selection of probiotics, revealed the ability to survive in vitro by many of the strains considered. This study proposed the best performing yeast strains selected among autochthonous sourdough yeasts based on their pro-technological, nutritional and functional traits to be used as starters for making sourdough baked goods or functional cereal-based beverages. Although some yeast strains combined several technological and nutritional traits, the association of more selected strains seemed to be a requisite to get optimal sourdough characteristics.


Genomic cis-regulatory logic: experimental and computational analysis of a sea urchin gene.
The genomic regulatory network that controls gene expression ultimately determines form and function in each species. The operational nature of the regulatory programming specified in cis-regulatory DNA sequence was determined from a detailed functional analysis of a sea urchin control element that directs the expression of a gene in the endoderm during development. Spatial expression and repression, and the changing rate of transcription of this gene, are mediated by a complex and extended cis-regulatory system. The system may be typical of developmental cis-regulatory apparatus. All of its activities are integrated in the proximal element, which contains seven target sites for DNA binding proteins. A quantitative computational model of this regulatory element was constructed that explicitly reveals the logical interrelations hard-wired into the DNA.