Steroid therapy for meconium aspiration syndrome in newborn infants.
Meconium aspiration syndrome may cause severe respiratory distress in the newborn infant, with an associated high morbidity and mortality. A chemical pneumonitis is believed to occur secondary to bile, bile acids and pancreatic secretions contained in meconium. It has therefore been hypothesised that corticosteroids may be of benefit in the management of this condition through their anti-inflammatory properties. The objective of this review was to determine whether steroid therapy for meconium aspiration syndrome decreases the morbidity and mortality associated with this condition without adverse effects. Searches were made of PREMEDLINE and MEDLINE from 1966 to April 2003, CINAHL back to 1982, Current Contents back to 1998, The Cochrane Central Register of Controlled Trials (CENTRAL, The Cochrane Library, Issue 1, 2003) and Oxford Database of Perinatal Trials. The search included cross-referencing of previous reviews, and a review of abstracts, conference and symposia proceedings published in Pediatric Research from 1993 to 2003. Randomised controlled trials and quasi-randomised trials comparing steroid treatment to no steroid treatment for neonates with meconium aspiration syndrome were considered for this review. The methodological quality of each trial was assessed independently by each author. Data were extracted, analysed and results reviewed independently by each author. Meta-analysis was performed with RevMan 4.2, using the fixed effects model. Mean difference (MD) and weighted mean differences (WMD) with 95% confidence intervals in brackets for continuous variables and Relative Risk (RR) with 95% confidence intervals for categorical data were reported. Three randomised controlled trials were identified. Two trials, by Wu 1999 (50 participants) and Yeh 1977 (35 participants), were included in the review. The trial by Davey 1995, as yet unpublished, was excluded from this review as insufficient information about methodology and results were available. On meta-analysis, there was no significant reduction in mortality [typical RR 0.95 (0.20, 4.58)]. A small but significant increase in duration of oxygen therapy was seen with the use of steroids [WMD 30.0 hours (8.4, 51.6)]. There was no significant difference in duration of hospital stay in the study by Wu 1999 [MD 0.00 days (-3.09, 3.09)]. Duration of mechanical ventilation was reported by Wu 1999 with no significant difference seen [MD -1.10 days (-2.79, 0.59)]. Incidence of air leak was reported by Yeh 1977 with no significant difference detected [RR 0.64 (0.18, 2.26)]. Long-term outcome was not reported in either of the two studies. At present, there is insufficient evidence to assess the effects of steroid therapy in the management of meconium aspiration syndrome. A further large randomised controlled trial assessing potential benefits and harm would be required to determine its role.


Monte Carlo based method for conversion of in-situ gamma ray spectra obtained with a portable Ge detector to an incident photon flux energy distribution.
A Monte Carlo based method for the conversion of an in-situ gamma-ray spectrum obtained with a portable Ge detector to photon flux energy distribution is proposed. The spectrum is first stripped of the partial absorption and cosmic-ray events leaving only the events corresponding to the full absorption of a gamma ray. Applying to the resulting spectrum the full absorption efficiency curve of the detector determined by calibrated point sources and Monte Carlo simulations, the photon flux energy distribution is deduced. The events corresponding to partial absorption in the detector are determined by Monte Carlo simulations for different incident photon energies and angles using the CERN's GEANT library. Using the detector's characteristics given by the manufacturer as input it is impossible to reproduce experimental spectra obtained with point sources. A transition zone of increasing charge collection efficiency has to be introduced in the simulation geometry, after the inactive Ge layer, in order to obtain good agreement between the simulated and experimental spectra. The functional form of the charge collection efficiency is deduced from a diffusion model.


Hypertension and obstructive sleep apnea.
Obstructive sleep apnea is a common disorder that is often unrecognized and underappreciated. Emerging evidence suggests that there is a causal link between obstructive sleep apnea and hypertension. This relationship appears to be independent of other comorbidities that have been previously linked to hypertension, such as obesity. The majority of studies support the contention that alleviation of sleep disordered breathing has a clinically significant beneficial impact on decreasing both nighttime and daytime blood pressure. A pathophysiologic basis for patients with sleep apnea having an increased risk for hypertension is not fully elucidated. However, there is consistent evidence that autonomic mechanisms are implicated. Sympathetic activation along with humoral responses to repetitive episodes of hypoxemia and apnea over the longer term may cause vasoconstriction, endothelial dysfunction, and possibly hypertension. Patients with sleep apnea are often obese and may be predisposed to weight gain. Hence, obesity may further contribute to hypertension in this patient population.


Fatal human herpesvirus type 1 infection in a white-handed gibbon (Hylobates lar).
This report documents a case of spontaneous, fatal, and likely recrudescent human herpesvirus type 1 (HHV-1) infection in a captive white-handed gibbon (Hylobates lar) confirmed by polymerase chain reaction (PCR). An approximately 44-year-old, captive, female, white-handed gibbon with a history of recurrent conjunctivitis and occasional seizures became acutely weak, disoriented, and ataxic. A postictal state was suspected by caretakers and veterinary staff, and euthanasia was ultimately elected because of lack of clinical improvement with supportive care. No significant abnormalities were detected at necropsy. Histologically, sections of cerebrum and midbrain contained minimal to mild, multifocal lymphoplasmacytic meningoencephalitis with numerous intranuclear viral inclusions within astrocytes and some neurons. The presumptive diagnosis of HHV-1-induced encephalitis was strengthened by nested PCR amplification of a segment of the herpesvirus DNA polymerase gene. Sequences from this region have been found to be unique to each herpesvirus species, thus identifying HHV-1 as the likely etiologic agent in this case. Positive HHV-1 serology from several years before the terminal episode suggested that the disease was most likely due to recrudescence of latent HHV-1 infection.


Plasma magnesium in adult asthmatics at the Lagos University Teaching Hospital, Nigeria.
To determine the plasma magnesium levels in adult asthmatics. Case control study. Lagos University Teaching Hospital, Lagos, Nigeria. Forty stable asthmatics and thirty five apparently healthy controls. Detailed history including drug history was obtained from patients and controls. Fasting venous blood was collected into heparinized tubes from both patients and controls. Plasma magnesium, albumin, calcium and phosphorus were measured using spectrophotometric end point methods. Mean (SD) plasma magnesium concentration of 0.93(0.24) mmol/L in the asthmatics was significantly lower compared to the controls which was 1.15(0.28) mmol/L, p<0.001. There was no significant difference in the plasma albumin, calcium and phosphorus levels between the patients and controls. Plasma magnesium levels correlated with the plasma albumin level (r=0.62) but did not correlate with the duration of disease, peak flow rate and age of patients. There was no gender difference in plasma magnesium levels in the asthmatics. Adult Nigerian asthmatics have lower plasma magnesium concentrations compared to healthy controls. This may be of use in future therapeutic management of asthma. A large scale multi-centre study needs to be carried out since this study was done in one hospital.


Sequestration of multiple RNA recognition motif-containing proteins by C9orf72 repeat expansions.
GGGGCC repeat expansions of C9orf72 represent the most common genetic variant of amyotrophic lateral sclerosis and frontotemporal degeneration, but the mechanism of pathogenesis is unclear. Recent reports have suggested that the transcribed repeat might form toxic RNA foci that sequester various RNA processing proteins. Consensus as to the identity of the binding partners is missing and whole neuronal proteome investigation is needed. Using RNA fluorescence in situ hybridization we first identified nuclear and cytoplasmic RNA foci in peripheral and central nervous system biosamples from patients with amyotrophic lateral sclerosis with a repeat expansion of C9orf72 (C9orf72+), but not from those patients without a repeat expansion of C9orf72 (C9orf72-) or control subjects. Moreover, in the cases examined, the distribution of foci-positive neurons correlated with the clinical phenotype (t-test P < 0.05). As expected, RNA foci are ablated by RNase treatment. Interestingly, we identified foci in fibroblasts from an asymptomatic C9orf72+ carrier. We next performed pulldown assays, with GGGGCC5, in conjunction with mass spectrometry analysis, to identify candidate binding partners of the GGGGCC repeat expansion. Proteins containing RNA recognition motifs and involved in splicing, messenger RNA nuclear export and/or translation were significantly enriched. Immunohistochemistry in central nervous system tissue from C9orf72+ patients with amyotrophic lateral sclerosis demonstrated co-localization of RNA foci with SRSF2, hnRNP H1/F, ALYREF and hnRNP A1 in cerebellar granule cells and with SRSF2, hnRNP H1/F and ALYREF in motor neurons, the primary target of pathology in amyotrophic lateral sclerosis. Direct binding of proteins to GGGGCC repeat RNA was confirmed in vitro by ultraviolet-crosslinking assays. Co-localization was only detected in a small proportion of RNA foci, suggesting dynamic sequestration rather than irreversible binding. Additional immunohistochemistry demonstrated that neurons with and without RNA foci were equally likely to show nuclear depletion of TDP-43 (χ(2) P = 0.75) or poly-GA dipeptide repeat protein inclusions (χ(2) P = 0.46). Our findings suggest two non-exclusive pathogenic mechanisms: (i) functional depletion of RNA-processing proteins resulting in disruption of messenger RNA splicing; and (ii) licensing of expanded C9orf72 pre-messenger RNA for nuclear export by inappropriate association with messenger RNA export adaptor protein(s) leading to cytoplasmic repeat associated non-ATG translation and formation of potentially toxic dipeptide repeat protein.


Exercise performance and skeletal muscles in patients with advanced Chagas disease.
This study was designed to evaluate the peripheral muscle metabolic and structural characteristics in patients with advanced Chagas disease (ChD), and whether they were related with exercise performance. Cross-sectional study. Outpatient cardiomyopathy clinic of a university hospital. We studied 11 stage II patients, 8 stage III patients, and 11 healthy volunteers. All patients underwent exercise testing and peripheral muscle biopsies. The muscles were also studied in control subjects. Muscle biopsy specimens were analyzed for histochemical characteristics. In six patients, the muscle was studied ultrastructurally. The data demonstrate more glycolytic and less oxidative capacity of the peripheral muscle in patients with advanced ChD (increased proportion of type IIb fibers, increased proportion of fibers with low nicotinamide adenine dinucleotide diaphorase activity, high proportion of darkly stained fibers for alpha-glycerophosphate dehydrogenase activity, and lower levels of citrate synthase). Many capillaries in patients with ChD had an abnormal aspect: they were either closed or showed a thicker wall. The ultrastructural study also showed fiber atrophy and abnormal capillaries even in patients with normal functional capacity. Some muscle characteristics (enzyme activity, mean cross-sectional area of the fiber, and capillarity) related with exercise parameters (anaerobic threshold, and peak oxygen pulse). These findings indicate that patients with advanced ChD have decreased oxidative capacity and a shift to anaerobic metabolism in the skeletal muscle. They also suggest that muscular abnormalities are related to oxygen delivery, which is probably reduced in part by the abnormal muscle microvasculature. Those changes could affect oxygen extraction, and therefore exercise tolerance in these patients.


Spread of parasites transported with their hosts: case study of two species of cattle tick.
Like all parasites, ticks can be spread easily along with their hosts. Ticks are obligate parasites of vertebrates, to which they attach themselves for varying periods of time, and are well-adapted to this mode of transport. Once the transport stage is complete and they have detached at destination, they are also able to wait several months for the arrival of a new host on which they will continue their life cycle. This leads to the establishment of a secondary tick population. Two tropical cattle tick species, Rhipicephalus microplus and Amblyomma variegatum, have perfected this strategy of colonisation and occupation of favourable zones. Rhipicephalus microplus, which originated from South and Southeast Asia, is highly specific for ungulates, and thanks to cattle movements it has spread throughout the tropical belt, apart from the remotest areas. Amblyomma variegatum, which originated in Africa, was transported to Madagascar and the Mascarene Islands, as well as to the West Indies, during the time of the Atlantic triangular trade. These two ticks are vectors of particularly serious cattle diseases: babesiosis and anaplasmosis in the case of R. microplus, and heartwater (cowdriosis) in the case of A. variegatum. Anticipated climate changes are likely to modify the potential geographical range of these two parasite species and numerous others. Even now there are still many areas of the Americas, Asia and Oceania into which A. variegatum has not yet spread, but which it would find favourable. It could be spread not only by the transport of cattle, but also by the migration of some of its other hosts, such as birds. Surveillance--and know-how--is needed to identify these parasites when they first appear and to rapidly contain new outbreaks. Efforts should be made to raise the awareness of livestock professionals about the risks of transporting cattle. Regulations should be implemented and precautions taken to avoid such artificial expansion of the range of ticks and the diseases they transmit.


Inspiratory Muscle Training in Patients with Heart Failure.
Prior systematic reviews and meta-analysis addressed that inspiratory muscle training (IMT) improved inspiratory muscle weakness, cardiorespiratory fitness and quality of life similar to conventional exercise training as a first alternative in deconditioned patients with heart failure (HF) lead to a better adaptation to posterior exercise training. The heterogeneity and variability in a wide range of new studies about this topic led to the necessity of an updated and comprehensive narrative review. The present review aimed to analyze and update the most relevant studies about IMT in patients who suffer from HF. A narrative review was carried out about IMT in HF patients including 26 experimental studies divided into 21 clinical trials and 5 quasi-experimental studies identified through database searching in PubMed, Cochrane and PEDro. There is enough evidence to state that IMT produces improvements in functional capacity of patients with HF. Nevertheless, there is not enough evidence to support that IMT could improve cardiovascular parameters, blood biomarkers or quality of life in these patients. Thus, IMT may be recommended to improve functional capacity in patients who suffer from HF; nevertheless, more evidence is needed regarding cardiovascular parameters, biomarkers and quality of life. Furthermore, mortality or HF hospitalization was not evaluated and most studies were not longer than 3 months. According to IMT protocols and study designs heterogeneity and mid-term follow-up, further investigations through high-quality long-term randomized clinical trials should be performed to achieve systematic reviews and meta-analysis to support strong evidence for IMT in HF patients.


Semen donors and STD screening.
The British Andrology Society recommends screening semen donors for sexually transmitted infections to minimise the risk of pathogen transmission to the mother and fetus. The aim was to review recent findings of semen donor screening and, if appropriate, recommend changes to the screening protocol. 175 consecutive men attending for STD screening between January 1992 and December 1995 who had been preselected by the Department of Obstetrics and Gynaecology as suitable semen donors. Retrospective review of case notes and group comparison of demographic and sexual history data. 11 men (6%) had evidence of infection, excluding CMV seropositivity, at their first STD screen. After semen donation, 109 men (63%) were rescreened and, of these, 12% had positive findings. Positive findings at initial screening were predicted by a history of more than one partner in the preceding 6 months (OR 7.11, 95% CI 1.66-30.4) but it did not predict rescreening findings. Other factors such as age, marital status, employment status or past STDs were not predictive for either screen. Less than 20% of initial volunteers meet the full criteria of high quality post-thaw semen, no transmissible genetic disorders, and no transmissible pathogens. Sexual history may predict but would not alone preclude all positive STD screening findings. It is essential that sequential STD screening of donors continues and that genitourinary physicians should be involved in this process. Validation of newer diagnostic techniques as screening tests in this setting is required.


Development of a prognostic model for predicting spontaneous singleton preterm birth.
To develop and validate a prognostic model for prediction of spontaneous preterm birth. Prospective cohort study using data of the nationwide perinatal registry in The Netherlands. We studied 1,524,058 singleton pregnancies between 1999 and 2007. We developed a multiple logistic regression model to estimate the risk of spontaneous preterm birth based on maternal and pregnancy characteristics. We used bootstrapping techniques to internally validate our model. Discrimination (AUC), accuracy (Brier score) and calibration (calibration graphs and Hosmer-Lemeshow C-statistic) were used to assess the model's predictive performance. Our primary outcome measure was spontaneous preterm birth at <37 completed weeks. Spontaneous preterm birth occurred in 57,796 (3.8%) pregnancies. The final model included 13 variables for predicting preterm birth. The predicted probabilities ranged from 0.01 to 0.71 (IQR 0.02-0.04). The model had an area under the receiver operator characteristic curve (AUC) of 0.63 (95% CI 0.63-0.63), the Brier score was 0.04 (95% CI 0.04-0.04) and the Hosmer Lemeshow C-statistic was significant (p<0.0001). The calibration graph showed overprediction at higher values of predicted probability. The positive predictive value was 26% (95% CI 20-33%) for the 0.4 probability cut-off point. The model's discrimination was fair and it had modest calibration. Previous preterm birth, drug abuse and vaginal bleeding in the first half of pregnancy were the most important predictors for spontaneous preterm birth. Although not applicable in clinical practice yet, this model is a next step towards early prediction of spontaneous preterm birth that enables caregivers to start preventive therapy in women at higher risk.


Fucoidan improves serum lipid levels and atherosclerosis through hepatic SREBP-2-mediated regulation.
Hyperlipidemia is associated with increased risk of the development of cardiovascular diseases. Although a great deal of attention has been paid to the hypolipidemic activity of fucoidan, complex polysaccharides from brown seaweeds, the underlying mechanism is still unclear. This study was performed to investigate whether and how fucoidan has lipid-lowering potential in poloxamer-407 (P407)-induced hyperlipidemic mice. Fucoidan treatment 2 h after acute administration of P407 in these mice significantly reduced serum total cholesterol, triglycerides, and LDL cholesterol levels, but increased the levels of HDL cholesterol. In HepG2 hepatocytes and the liver, fucoidan decreased the expression of FAS and ACC mRNA with no or only a moderate inhibitory effect on SREBP-1c mRNA expression. Furthermore, fucoidan attenuated the hepatic expression of mature SREBP-2 protein with a subsequent decrease in hepatic HMG-CoA reductase mRNA expression and an increase in hepatic LDL receptor mRNA expression. In addition, atherosclerotic lesions in the aorta of chronically P407-treated mice were also reduced by fucoidan. These findings indicate that fucoidan improves serum lipid levels by regulating the expression of key enzymes of cholesterol and triglyceride syntheses in the liver through modulation of SREBP-2.


Electrochemical heparin sensing at liquid/liquid interfaces and polymeric membranes.
The monitoring of heparin and its derivatives in blood samples is important for the safe usage of these anticoagulants and antithrombotics in many medical procedures. Such an analytical task is, however, highly challenging due to their low therapeutic levels in the complex blood matrix, and it still relies on classical, indirect, clot-based assays. Here we review recent progress in the direct electrochemical sensing of heparin and its analogs at liquid/liquid interfaces and polymeric membranes. This progress has been made by utilizing the principle of electrochemical ion transfer at the interface between two immiscible electrolyte solutions (ITIES) to voltammetrically drive the interfacial transfer of polyanionic heparin and monitoring the resulting ionic current as a direct measure of heparin concentration. The sensitivity, selectivity, and reproducibility of the ion-transfer voltammetry of heparin are dramatically enhanced compared to those of traditional potentiometry. This voltammetric principle was successfully applied for the detection of heparin in undiluted blood samples, and was used to develop highly sensitive ion-selective electrodes based on thin polymeric membranes that are intended for analytical applications beyond heparin detection. The mechanism of heparin recognition and transfer at liquid/liquid interfaces was assessed quantitatively via sophisticated micropipet techniques, which aided the development of a powerful ionophore that can extract large heparin molecules into nonpolar organic media. Moreover, the reversible potentiometric detection of a lethal heparin-like contaminant in commercial heparin preparations was achieved through the use of a PVC membrane doped with methyltridodecylammonium chloride, which enables charge density dependent polyanion selectivity.


First 4-years orthognathic surgery audit of a Saudi teaching hospital.
The study represents a retrospective audit of the first four years of orthognathic surgery carried out at a Saudi teaching hospital. The results showed that orthognathic surgery was the most common oral and maxillofacial surgery performed in that particular hospital, which is one of two teaching hospitals covered by the division of oral and maxillofacial surgery, faculty of dentistry, King Saud University in Riyadh, Kingdom of Saudi Arabia. The variation in the patients characteristics, treatment provided, and final outcome is presented.


Grey-Matter Metabolism in Relation with White-Matter Lesions in Older Hypertensive Patients with Subjective Memory Complaints: A Pilot Voxel-Based Analysis Study.
This study aimed at assessing the changes in brain metabolism related to white-matter magnetic resonance (MR) hyperintensities of presumed vascular origin, with a voxel-based quantitative analysis of (18F)-fluorodesoxyglucose positron emission tomography (FDG-PET) imaging. Sixty older hypertensive patients with subjective memory complaints (75 ± 5 years, 34 women) were prospectively referred to FDG-PET and MRI brain imaging. The Statistical Parametric Mapping software was used to assess the correlation between brain distribution of FDG and white-matter hyperintensities assessed by the Fazekas score on MRI images. The Fazekas score was inversely related to FDG uptake, independently of age and gender, within 14 Brodmann areas located mainly in the frontal lobe but also in certain limbic, insular and temporal areas. This relationship was also found to be largely independent of the volume of grey matter expressed in percentage of cranial volume, an index of atrophy. White-matter MR hyperintensities of presumed vascular origin are cross-sectionally associated with a lower grey-matter metabolism, mainly but not only within frontal areas and independently of age, gender and grey-matter atrophy.


Characteristics of new patients at Illinois cleft palate teams.
The Illinois Association of Craniofacial Teams (IACT) surveyed all new patient visits to member teams in an effort to assess the needs of patients in Illinois. The survey determined the number of patients, their age at initial visit, the location and status of their defect, and their history of team visits. Four hundred eighty new patient visits were recorded. The number of patients was substantially lower than expected, and the large majority of older children seeing a team for the first time had repaired defects. It is concluded that the main opportunity for improving the quality of care for patients with clefts in Illinois may be in increasing the use of cleft palate teams.


Transcranial color duplex sonography in childhood and adolescence. Age dependence of flow velocities and waveform parameters.
Transcranial color duplex sonographic examinations in children and adolescents without cerebrovascular disease were evaluated retrospectively. Flow velocities and waveform parameters were determined and their side-to-side differences and age dependence analyzed and, finally, compared with analogous data of a previously described group of healthy adults. With a 2.0-MHz sector transducer of a computed sonographic system, the anterior, middle, and posterior cerebral arteries were examined in 64 children and adolescents between 1.5 and 17.5 years of age. Angle-corrected systolic peak, end-diastolic maximum, time-averaged, and time-averaged maximum velocities, and the resistance, pulsatility, and spectral broadening indexes were determined in all vessels. Mean +/- SD values for time-averaged maximum velocity (and time-averaged velocity) were 92.2 +/- 13.0, 79.9 +/- 17.7, and 63.9 +/- 13.6 (55.3 +/- 11.7, 40.4 +/- 10.4, and 34.2 +/- 9.2) cm/s, respectively, in the middle, anterior, and posterior cerebral arteries in children under 10 years of age; values were 83.2 +/- 11.9, 69.4 +/- 13.8, and 55.6 +/- 10.1 (50.8 +/- 9.0, 39.9 +/- 10.5, and 33.1 +/- 6.3) cm/s, respectively, in children 10 years of age and older. Time-averaged maximum velocity decreased significantly with age in all vessels (P < .001). Although time-averaged velocity did not change significantly during childhood and adolescence, a clear decline occurred from adolescence to adulthood (P < .0001 in the middle and posterior cerebral arteries; P < .01 in the anterior cerebral artery). The spectral broadening index decreased significantly from childhood to adolescence in the anterior and posterior cerebral arteries (P < .0001). The resistance and pulsatility indexes remained stable throughout childhood. Transcranial color duplex sonography allows angle-corrected measurements of "true" flow velocities in basal cerebral arteries. Additional determination of time-averaged velocity permits more detailed evaluation of flow characteristics for all age groups. The transcranial color duplex technique may provide deeper insights on normal cerebral perfusion and its disorders.


Design, synthesis and antimicrobial evaluation of novel 1,3-oxazolidin-2- one derivatives.
A series of novel derivatives of 1,3-oxazolidin-2-one 12a-12n has been synthesized starting from 4-nitro-(L)- phenylalanine by involving five-step reaction sequence. All the compounds were screened for their in vitro antibacterial activity against four pathogenic bacterial strains namely, Staphylococcus aureus, Bacillus subtilis (Gram-positive), Escherichia coli, Pseudomonas aeruginosa (Gram-negative) and in vitro antifungal activity against two pathogenic fungal strains namely, Candida albicans and Saccharomyces cerevisiae. All the synthesized compounds showed activity against Gram-positive bacteria. Compounds 12c and 12l exhibited maximum antibacterial activity against Gram-positive bacteria. However, against Gram-negative bacteria only five of screened compounds were found to be active. Compounds 12c and 12i displayed best antifungal activity against the tested fungi. Docking studies were carried out in order to gain insight into the mechanism of action and the binding mode of these compounds. These studies were in agreement with the biological data.


Spectroscopic investigation of the anticancer alkaloid piperlongumine binding to human serum albumin from the viewpoint of drug delivery.
Piperlongumine (PL) is a very promising natural agent with a high potential for cancer treatment. To overcome the poor water solubility of PL, there is a need to develop a novel water-soluble formulation in which PL is non-covalently bound to human serum albumin (HSA). PL binding to HSA was studied by various spectroscopic techniques under simulated physiological conditions. Spectroscopic evidence showed that the interaction of PL with HSA could form a PL-HSA complex. The binding constant (Ka ) values increased with increasing temperature, and a similar dependence was observed for the number of binding sites (n) values. The number of PL molecules bound to HSA reached 8.1 when the temperature was raised to 308 K. Thermodynamic calculation results suggested that the binding reaction occurred spontaneously but was an entropy-driven process, and hydrophobic forces played a major role in stabilizing the complex. Furthermore, PL binding induced conformational and microenvironmental changes in HSA. Displacement studies indicated that PL and warfarin had separate binding regions in site I. Therefore, it would be possible to develop a novel water-soluble formulation involving PL and HSA. This study may provide some valuable information in terms of improving the poor water solubility of PL.


The role of the MRTF-A/SRF pathway in ocular fibrosis.
Tissue contraction and fibrosis are major causes of morbidity in the human body. In the eye in particular, fibrosis and scarring are responsible for the pathogenesis or failure of treatment of all major blinding diseases, with postoperative wound healing responses posing a major problem for most ocular surgery on a worldwide scale. This is one of the largest areas of unmet need in ophthalmology, with currently no antifibrotic treatments available clinically. This review focuses on the ubiquitous myocardin-related transcription factor/serum response factor (MRTF-A/SRF) transcription pathway as a potential novel therapeutic target in fibrotic eye diseases. It describes how the MRTF-A/SRF pathway is intricately linked to all the key regulators and pathways in ocular fibrosis, and how it could potentially lead to a new avenue of antifibrotic therapies in the future.


Early dynamic fate changes in haemogenic endothelium characterized at the single-cell level.
Haematopoietic stem cells (HSCs) are the founding cells of the adult haematopoietic system, born during ontogeny from a specialized subset of endothelium, the haemogenic endothelium (HE) via an endothelial-to-haematopoietic transition (EHT). Although recently imaged in real time, the underlying mechanism of EHT is still poorly understood. We have generated a Runx1 +23 enhancer-reporter transgenic mouse (23GFP) for the prospective isolation of HE throughout embryonic development. Here we perform functional analysis of over 1,800 and transcriptional analysis of 268 single 23GFP(+) HE cells to explore the onset of EHT at the single-cell level. We show that initiation of the haematopoietic programme occurs in cells still embedded in the endothelial layer, and is accompanied by a previously unrecognized early loss of endothelial potential before HSCs emerge. Our data therefore provide important insights on the timeline of early haematopoietic commitment.


Regulation of GluR1 by the A-kinase anchoring protein 79 (AKAP79) signaling complex shares properties with long-term depression.
Second messengers regulate synaptic plasticity by influencing the balance between kinase and phosphatase activity. One target of this balance is the phosphorylation state of the AMPA receptor glutamate receptor 1 (GluR1) subunit. Hippocampal long-term depression (LTD) is a calcium-dependent downregulation of synaptic AMPA receptor currents associated with dephosphorylation of Ser845, a cAMP-dependent protein kinase (PKA) site on GluR1. Recruitment of kinases and phosphatases to the AMPA receptor might enable modulation of AMPA receptor function. The neuronal A-kinase anchoring protein AKAP79/150 interacts with PKA and the calcium-dependent protein phosphatase PP2B and is linked to the AMPA receptor GluR1 subunit by synapse-associated protein 97 (SAP97), a membrane-associated guanylate kinase family protein. Here we demonstrate that AKAP79 not only promotes basal phosphorylation of Ser845 but also confers a calcium- and PP2B-mediated downregulation to GluR1 receptor currents. This AKAP79-dependent downregulation is contingent on the local presence of PKA, Ser845 of GluR1, and a PDZ (postsynaptic density 95/Discs large/zona occludens 1)-domain interaction between GluR1 and SAP97, all of which support basal phosphorylation of the receptor. These findings suggest that the AKAP79 signaling complex is sufficient to couple intracellular calcium levels to the PKA phosphorylation state of GluR1. Thus, the integration of intracellular signals relevant for LTD may be transduced to GluR1 by the AKAP79 signaling complex.


Molecular characterization of Cryptosporidium in bats from Yunnan province, southwestern China.
The present study was conducted to investigate the prevalence and genotypes of Cryptosporidium in bats. A total of 247 bats, belonging to Rhinolophus sinicus , Rousettus leschenaultia, Aselliscus stoliczkanus , and Hipposideros fulvus , were collected in Yunnan Province, Southwestern China, and the intestinal tissues were examined for Cryptosporidium infection by PCR amplification of the small subunit ribosomal RNA (SSU rRNA). The overall infection rate was 7.7% (95% CI, 4.5 to 11.0%), with R. sinicus having the highest level at 9.5% (95% CI, 2.8 to 16.1%) followed by A. stoliczkanus at 7.8% (95% CI, 2.2 to 18.9%), H. fulvus at 7.2% (95% CI, 1.1 to 13.4%), and R. leschenaultia at 5.7% (95% CI, 1.2 to 15.7%). DNA sequence and phylogenetic analyses based on SSU rRNA revealed the presence of 2 novel genotypes, designated as Cryptosporidium bat genotype I in A. stoliczkanus and R. sinicus and Cryptosporidium bat genotype II in R. leschenaultia, R. sinicus , and H. fulvus . This is the first report of Cryptosporidium genotypes in bats. Further biological and genetic characterization is needed to determine the relationship of the 2 novel genotypes to established Cryptosporidium species-genotypes.


Kinetochore-dependent microtubule rescue ensures their efficient and sustained interactions in early mitosis.
How kinetochores regulate microtubule dynamics to ensure proper kinetochore-microtubule interactions is unknown. Here, we studied this during early mitosis in Saccharomyces cerevisiae. When a microtubule shrinks and its plus end reaches a kinetochore bound to its lateral surface, the microtubule end attempts to tether the kinetochore. This process often fails and, responding to this failure, microtubule rescue (conversion from shrinkage to growth) occurs, preventing kinetochore detachment from the microtubule end. This rescue is promoted by Stu2 transfer (ortholog of vertebrate XMAP215/ch-TOG) from the kinetochore to the microtubule end. Meanwhile, microtubule rescue distal to the kinetochore is also promoted by Stu2, which is transported by a kinesin-8 motor Kip3 along the microtubule from the kinetochore. Microtubule extension following rescue facilitates interaction with other widely scattered kinetochores, diminishing long delays in collecting the complete set of kinetochores by microtubules. Thus, kinetochore-dependent microtubule rescue ensures efficient and sustained kinetochore-microtubule interactions in early mitosis.


Impact of thawing on RNA integrity and gene expression analysis in fresh frozen tissue.
Biobanks of fresh, unfixed human tissue represent a valuable source for gene expression analysis in translational research and molecular pathology. The aim of this study was to evaluate the impact of thawing on RNA integrity and gene expression in fresh frozen tissue specimens. Portions of snap frozen tonsil tissue, unfixed or immersed in RNAlater, were thawed at room temperature for 0 minute, 5 minutes, 30 minutes, 45 minutes, 1 hour, 3 hours, 6 hours, and 16 hours before RNA extraction. Additionally, tonsil tissue underwent repetitive freezing and thawing cycles. RNA integrity was analyzed by microchip gel electrophoresis and gene expression by quantitative real-time polymerase chain reaction for selected genes (FOS, TGFB1, HIF1A, BCL2, and PCNA). Minimal RNA degradation was detected after 30 minutes of thawing in unfixed samples. This degradation was accompanied by relevant changes in gene expression for FOS and BCL2 at 45 minutes. Modified primer design or the use of different housekeeping genes could not rectify the changes for FOS. Repetitive thawing cycles had similar effects on RNA integrity. The incubation of the tissue in RNAlater efficiently prevented RNA degradation. In conclusion, degradation of RNA in frozen tissue occurs first after several minutes of thawing. Already minimal decrease in RNA quality may result in significant changes in gene expression patterns in clinical tissue samples.


Cardiac β1-adrenoceptor expression in two stress-induced cardiomyopathy-related deaths.
Stress-induced cardiomyopathy (SICM) is characterized by transient systolic dysfunction of the apical and/or midventricular myocardial segments in the absence of obstructive coronary artery disease and is unique in that it can manifest itself after acute emotional stress. Excessive amounts of catecholamines released from sympathetic nerve endings as well as from the adrenal medulla under stressful conditions are considered to produce intracellular Ca(2+) overload and cardiac dysfunction through the β(1)-adrenoceptor signal transduction pathway. We describe the clinical and pathomorphological findings in two stress-induced cardiomyopathy fatal cases. Levels of catecholamines and their metabolites in urine samples were assessed too. Morphological patterns seen in SICM result from the complex interplay between sympathetic innervations, β-receptor density and function and catecholamine sensitivity.


Detection of candidal antigens in autoimmune polyglandular syndrome type I.
Autoimmune polyglandular syndrome type I (APS I) is associated with chronic mucocutaneous candidiasis. To characterize the antibody responses in this subgroup of Candida albicans infections, we screened a candidal cDNA expression library with patient sera and found four cDNA clones encoding the immunopositive proteins enolase, heat shock protein 90, pyruvate kinase, and alcohol dehydrogenase. The reactivity to these antigens was studied further by immunoprecipitation assays with in vitro-transcribed and -translated proteins. Analysis of sera from 44 APS I patients showed that the highest antibody reactivity was found with enolase (80% of patients reactive), but significant serological responses were also found with heat shock protein 90 (67%), pyruvate kinase (62.5%), and alcohol dehydrogenase (64%). Overall, 95.5% of patients had detectable antibodies to at least one of these proteins. The cDNAs of enolase and heat shock protein 90 were also expressed in Escherichia coli and studied by immunoblotting. Again, 84% of sera reacted with enolase, whereas 44% of sera reacted with heat shock protein 90. A good correlation between the two methods was found for both enolase (r = 0.86; n = 58; P < 0.001) and heat shock protein 90 (r = 0.71; n = 56; P < 0.001). Our results indicate that the four abundant candidal proteins are the major antigens and can be used as accurate markers of candidiasis in APS I patients. The immunoprecipitation assay described here is particularly useful for the rapid analysis of a large number of samples.


Acidic pH or salt treatment can convert soluble antibody aggregates in culture harvest into monomers and improve Protein A chromatography step yield.
While purifying a regular monospecific antibody, we found that the Protein A step yield was much lower than expected. Further studies revealed that the antibody formed large-size aggregates that did not bind to the Protein A resin, hence leading to dropped recovery. In an attempt to solve this low yield issue, we found that mildly acidic pH or ammonium sulfate treatment can partially convert the aggregates into monomers. In addition, when acidic pH treated culture harvest was processed by Protein A chromatography, the yield was restored to the normal range, suggesting that the monomers recovered from aggregates regained Protein A binding capability. Thus, low pH treatment of culture harvest can be potentially used as a general approach for improving Protein A step yield in cases where non-binding antibody aggregates are formed through noncovalent interactions.


Activation of the oxidative stress pathway by HIV-1 Vpr leads to induction of hypoxia-inducible factor 1alpha expression.
The detection of biomarkers of oxidative stress in brain tissue and cerebrospinal fluid of patients with human immunodeficiency virus, type 1 (HIV)-associated dementia indicates the involvement of stress pathways in the neuropathogenesis of AIDS. Although the biological importance of oxidative stress on events involved in AIDS neuropathogenesis and the HIV-1 proteins responsible for oxidative stress remain to be elucidated, our results point to the activation of hypoxia-inducible factor 1 (HIF-1) upon HIV-1 infection and its elevation in brain cells of AIDS patients with dementia. HIF-1 is a transcription factor that is responsive to oxygen. Under hypoxic conditions, HIF-1alpha becomes stable and translocates to the nucleus where it dimerizes with aryl hydrocarbon receptor nuclear translocator and modulates gene transcription. Activation of HIF-1 can also be mediated by the HIV-1 accessory protein Vpr. In addition, cellular components, including reactive oxygen species, contribute to the induction of HIF-1alpha. Our results show that Vpr induces reactive oxygen species by increasing H(2)O(2) production, which can contribute to HIF-1alpha accumulation. Interestingly, increased levels of HIF-1alpha stimulated HIV-1 gene transcription through HIF-1 association with HIV-1 long terminal repeat. These observations point to the existence of a positive feedback interplay between HIF-1alpha and Vpr and that, by inducing oxidative stress via activation of HIF-1, Vpr can induce HIV-1 gene expression and dysregulate multiple host cellular pathways.


Making the PACS workstation a browser of image processing software: a feasibility study using inter-process communication techniques.
To enhance the functional expandability of a picture archiving and communication systems (PACS) workstation and to facilitate the integration of third-part image-processing modules, we propose a browser-server style method. In the proposed solution, the PACS workstation shows the front-end user interface defined in an XML file while the image processing software is running in the background as a server. Inter-process communication (IPC) techniques allow an efficient exchange of image data, parameters, and user input between the PACS workstation and stand-alone image-processing software. Using a predefined communication protocol, the PACS workstation developer or image processing software developer does not need detailed information about the other system, but will still be able to achieve seamless integration between the two systems and the IPC procedure is totally transparent to the final user. A browser-server style solution was built between OsiriX (PACS workstation software) and MeVisLab (Image-Processing Software). Ten example image-processing modules were easily added to OsiriX by converting existing MeVisLab image processing networks. Image data transfer using shared memory added <10ms of processing time while the other IPC methods cost 1-5 s in our experiments. The browser-server style communication based on IPC techniques is an appealing method that allows PACS workstation developers and image processing software developers to cooperate while focusing on different interests.


Illness, disability, and drugs among 25 to 75 year olds living at home.
A survey in a London borough showed that 15% of adults living at home were restricted in one or more areas of their lives because of illness. A sample of these adults aged 25 to 75 years was interviewed using a validated medical questionnaire, and the severity of their restrictions was also assessed using a separate instrument. Many symptoms were found which had not been reported to a doctor and many were not being treated. When the disability scores were regressed on symptoms classified as reported to a doctor, unreported, or absent, with a few exceptions it was the reported symptoms that were significantly associated with disability. Similarly, when symptoms were classified as treated (by doctor or respondent), untreated, or absent, treated symptoms were associated with disability. Some disabling symptoms were similar to the effects, mainly adverse, of commonly prescribed drugs, and these symptoms were reported more frequently by respondents taking the possible offending drug than by those not taking the drug. It appears that making general practitioners aware of unreported and untreated symptoms among their 25 to 75 year old patients will not reduce the overall level of disability in the community. However, the iatrogenic component of disability needs to be studied further.


Orbital Inflammatory Syndrome and Anterior Uveitis: A Case Series.
To describe four cases of orbital inflammatory syndrome (OIS) with associated anterior uveitis that have presented within 2 years to our practice. Charts of patients diagnosed with OIS from June 2013 to May 2015 were reviewed. Four patients, three children and one adult, presented with orbital swelling, pain, and varying degrees of vision loss. Treatment with intravenous methylprednisolone resulted in significant symptomatic improvement in all cases initially; when symptoms recurred, the patients had evidence of anterior uveitis. With continued systemic therapy and the addition of topical prednisolone, the patients all achieved control of their uveitis and OIS and are well controlled with regular outpatient follow-up. Reports of OIS-associated with uveitis are relatively rare. The presentation of three pediatric patients and one adult patient to the same practice with OIS and secondary uveitis within a 2-year period may indicate that the association is underreported.


Characterization of p73 functional domains necessary for transactivation and growth suppression.
p73, a p53 family member, is highly similar to p53 in both structure and function. Like p53, the p73 protein contains an N-terminal activation domain, a DNA-binding domain, a tetramerization domain, and several PXXP motifs. Previously, we and others have shown that some functional domains in p53, such as the DNA-binding and tetramerization domains, are required for inducing both cell cycle arrest and apoptosis whereas others, such as the second activation domain, the proline-rich domain, and the C-terminal basic domain, are only required for inducing apoptosis. To determine the activity of p73 functional domains, we have generated stable inducible cell lines that express p73beta and various mutants deficient in one or more functional domains. We found that in addition to the DNA-binding domain, p73-mediated growth suppression requires the N-terminal activation domain and the tetramerization domain. However, unlike p53, p73-mediated apoptosis does not require the region adjacent to the activation domain or the entire C-terminal region. Interestingly, while the N- or the C-terminal PXXP motifs are dispensable for p73 function, deletion of both the N- and the C-terminal PXXP motifs renders p73 inactive in transactivation. In addition, we found that substitution of two conserved tandem hydrophobic residues with two hydrophilic ones, which can abrogate the activity of the first activation domain in p53, has no effect on p73 transcriptional activity. Together, we showed that the p73 protein has its own unique determinants for transactivation and growth suppression.


Regulation of gene expression downstream of a novel Fgf/Erk pathway during Xenopus development.
Activation of Map kinase/Erk signalling downstream of fibroblast growth factor (Fgf) tyrosine kinase receptors regulates gene expression required for mesoderm induction and patterning of the anteroposterior axis during Xenopus development. We have proposed that a subset of Fgf target genes are activated in the embyo in response to inhibition of a transcriptional repressor. Here we investigate the hypothesis that Cic (Capicua), which was originally identified as a transcriptional repressor negatively regulated by receptor tyrosine kinase/Erk signalling in Drosophila, is involved in regulating Fgf target gene expression in Xenopus. We characterise Xenopus Cic and show that it is widely expressed in the embryo. Fgf overexpression or ectodermal wounding, both of which potently activate Erk, reduce Cic protein levels in embryonic cells. In keeping with our hypothesis, we show that Cic knockdown and Fgf overexpression have overlapping effects on embryo development and gene expression. Transcriptomic analysis identifies a cohort of genes that are up-regulated by Fgf overexpression and Cic knockdown. We investigate two of these genes as putative targets of the proposed Fgf/Erk/Cic axis: fos and rasl11b, which encode a leucine zipper transcription factor and a ras family GTPase, respectively. We identify Cic consensus binding sites in a highly conserved region of intron 1 in the fos gene and Cic sites in the upstream regions of several other Fgf/Cic co-regulated genes, including rasl11b. We show that expression of fos and rasl11b is blocked in the early mesoderm when Fgf and Erk signalling is inhibited. In addition, we show that fos and rasl11b expression is associated with the Fgf independent activation of Erk at the site of ectodermal wounding. Our data support a role for a Fgf/Erk/Cic axis in regulating a subset of Fgf target genes during gastrulation and is suggestive that Erk signalling is involved in regulating Cic target genes at the site of ectodermal wounding.


Patient outcome study of concurrent chemoradiation, external beam radiotherapy, and high-dose rate brachytherapy in locally advanced carcinoma of the cervix.
A regimen of concurrent chemoradiation for definitive treatment of cervical cancer is widely used. This retrospective review has been conducted to determine the outcomes and late toxic effect associated with the specific regimen of whole-pelvic external beam radiotherapy of 45 Gy in 25 fractions with parametrial boosts of 5.4 or 9 Gy and HDR brachytherapy (BT) of 30 Gy in 5 fractions to point A delivered by tandem and ring. This protocol is accepted by the Gynecological Oncology Group and endorsed by the American Brachytherapy Society, but no late toxic effect data have been reported. The electronic records of sequential patients treated definitively at the Sunnybrook Odette Cancer Centre between January 2006 and December 2008 were reviewed. Patient-, tumor-, and treatment-related details (including external beam radiotherapy, BT, and chemotherapy) were obtained. Outcome measures included disease-free status, dates and sites of first recurrence, survival, and grade 3/4 late toxic effect results (Common Terminology Criteria Adverse Events 3.0 criteria). Exclusion criteria were no follow-up or a planned alternative regimen. One hundred twenty-two patients (+11 excluded) were treated with a median follow-up of 18 months from diagnosis. The actuarial 2-year disease-free survival rate was 70%. The median time to recurrence was 8 months (range, 2-22 months). The median time to toxic effect was 10 months (range 4-27 months). Grade 3/4 toxic effect was observed in 13 patients (11%). The actuarial grade 3/4 toxic effect rate at 2 years was 14%. Despite a relatively short follow-up, the toxicity of this regimen seems high compared with other retrospective series, although pelvic control is good. Consideration should be given to a reduction in BT dose alternatively when feasible image-guided BT may allow maintenance of tumor dose with reduced dose to organs at risk.


Frequency and quality of mid-afternoon snack among Spanish children.
There are increasing concerns about the types of foods and beverages consumed by children during mid-afternoon snack in Spain. To describe the frequency and quality of the mid-afternoon snack in a representative sample of Spanish children. A total of 1,233 younger (three to six years) and 1,618 older (seven to twelve years) children participated in the study. The report of food and beverages consumed per week during the mid-afternoon snack meal (merienda)was obtained by an online platform. An 84.4% of younger and 78.3% of older children were mid-afternoon snack consumers. Regular consumers were more likely to be from the South and the Centre regions. Sandwich was the most consumed food item among younger (49.5%) and older (59.7%) consumers; 46.5% of mid-afternoon snacks contained two food items among younger children, and 50.1% of mid-afternoon snacks contained one food item among older children. "Fruit", "milk and biscuits", and "fruit, biscuits and juices" were the most consumed combinations when considering one, two and three or more food items during one-time mid-afternoon occasion, respectively. The frequency of mid-afternoon snack occasions and the number and quality of food items consumed during one-time mid-afternoon snack occasion decrease with age. Spanish children should increase the frequency of mid-afternoon snack consumption and include healthy options in their menus. Longitudinal and intervention studies are needed to address the health impact of snacking and how to use this meal occasion to balance children's diets.


Prenatal and postnatal changes in the content and species of ferritin in rat liver.
The iron and ferritin content of rat liver and the species of ferritin present were examined from 4 days before to 3 weeks after birth. 1. Total iron and ferritin iron accumulated rapidly during the last days of gestation and from the second postnatal day underwent a steady depletion. 2. The amount of iron deposited before birth in the liver of each pup varied inversely with litter size and could be increased moderately by injection of iron into the mother before mating. 3. Intraperitoneal injection of iron 1 day after birth doubled the concentration of total iron, ferritin iron and ferritin protein in the liver over the next 24h, but at 3 weeks after birth it raised the very low concentrations of iron and ferritin severalfold. 4. As shown by electrophoretic migration, ferritin and dissociated ferritin subunits prepared from the livers of rats from 4 days before to 3 weeks after birth differed from those of adult liver ferritin and were indistinguishable from those of adult kidney and spleen ferritin. Treatment with iron at 3 weeks of age induced formation of a ferritin with electrophoretic properties resembling those of adult liver. It is concluded that iron given at this stage of development may activate the genetic cistron for adult liver ferritin.


Social synchronization of circadian rhythms with a focus on honeybees.
Many animals benefit from synchronizing their daily activities with conspecifics. In this hybrid paper, we first review recent literature supporting and extending earlier evidence for a lack of clear relationship between the level of sociality and social entrainment of circadian rhythms. Social entrainment is specifically potent in social animals that live in constant environments in which some or all individuals do not experience the ambient day-night cycles. We next focus on highly social honeybees in which there is good evidence that social cues entrain the circadian clocks of nest bees and can override the influence of conflicting light-dark cycles. The current understanding of social synchronization in honeybees is consistent with self-organization models in which surrogates of forager activity, such as substrate-borne vibrations and colony volatiles, entrain the circadian clocks of bees dwelling in the dark cavity of the nest. Finally, we present original findings showing that social synchronization is effective even in an array of individually caged callow bees placed on the same substrate and is improved for bees in connected cages. These findings reveal remarkable sensitivity to social time-giving cues and show that bees with attenuated rhythms (weak oscillators) can nevertheless be socially synchronized to a common phase of activity. This article is part of the theme issue 'Synchrony and rhythm interaction: from the brain to behavioural ecology'.


Association of elevated IGFBP-1 with increased IGF-II concentration in patients with carcinoma of the liver.
Insulin-like growth factors (IGFs) are mitogens for numerous types of cells including cancer cells. The aim of this work was to analyze some of the components of the IGF system to assess which could be potential clinical biomarkers for monitoring patients diagnosed with liver cancer. Compared to healthy persons, patients with liver cancer had a lower concentration of IGF-I and a higher concentration of IGFBP-1, whereas the concentrations of IGF-II and IGFBP-3 remained unchanged. The IGF-I:IGFBP-3 ratio decreased in patients with cancer, while the IGF-II:IGFBP-1 ratio was not altered. Patients with primary carcinoma and those scheduled for surgery had lower IGF-I and higher IGF-II and IGFBP-1 concentrations than patients with secondary carcinoma and those not eligible for surgery. It may be postulated that a liver with primary cancer is induced to increase IGF-II and IGFBP-1 synthesis more than a liver involved in metastatic response. Similarly, in patients eligible for liver surgery an increase in IGF-II may reflect a gradual change in the concentration associated with a different stage of disease. As increased synthesis of certain IGFBPs is necessary to compensate decreased production of the others or increased IGF production, determination of serum IGF-II, IGFBP-1 and their ratio may aid in estimating the compensatory capacity of the liver affected by cancer.


Evaluation of the effect of ultraviolet light polymerized clear adhesive on shear and tensile bond strength of heat cure denture base resin to the cobalt-chromium retentive minor connector: An in vitro study.
The aim of this study is to evaluate the efficacy of new material ultraviolet (UV) light polymerized clear adhesive on shear and tensile bond strength of heat cure denture base resin (Polymethylmethacrylate (PMMA)) to cobalt-chromium (Co-Cr) retentive minor connector. Comparative evaluation- In-vitro study. Sixty samples of Co-Cr plates mimicking minor connectors were fabricated. Thirty samples were coated with new material UV light polymerized clear adhesive and cured under UV light source for 10 min. In gun-metal flask, metal plates were placed in the lower compartment over it. Heat cure acrylic resin was packed in the dough stage with the help of clamps and processed according to the manufacturer's instructions. Samples were kept in artificial saliva for 90 days. Shear and tensile bond strengths were calculated of each sample with a Universal testing machine, and results were statistically analyzed. Type of bond failure was observed for each sample under stereomicroscope. Unpaired t test. Tensile bond strength, as well as shear bond strength, showed that 0.93 N and 1.64 N respectively for without application of new adhesive was more as compared to that of samples with the application of new adhesive which is 0.75 N and 1.54 N respectively. Bond failure was found to be an adhesive failure in resin-metal interface. Excellent bonding seen between the new adhesive and acrylic interface but limited effect on the metal interface. To increase bond strength between metal and resin interface, some surface treatment with the metal surface is needed to increase the bonding of the new adhesive to the metal surface.


Plasmin accelerates platelet-dependent prothrombinase formation without activating the platelets.
Patients with acute myocardial infarction who undergo thrombolytic therapy may shortly thereafter present evidence for increased platelet activation and thrombin activity, and recurrent thrombosis. This study investigated whether plasmin activates platelets and prothrombin in recalcified platelet-rich plasma (RPRP) to cause (at least in part) these side-effects of thrombolytic therapy. Plasmin (0.1 and 1.0 CU/ml) addition to RPRP with microM r-tick anticoagulant peptide (the latter a factor Xa inhibitor which abrogates prothrombin activation by prothrombinase at the concentration used) resulted in no change in the concentration of prothrombin fragment 1 + 2, or in the expression of GMP-140, the resting and activated GP IIb-IIIa conformers, and GPIb on platelets. Thus, plasmin neither activates platelets nor prothrombin in RPRP. However, plasmin accelerated platelet activation and secretion, and prothrombin fragment 1 + 2 production in RPRP. When combined with 1 microM r-tick anticoagulant peptide and 1 or 10 mM alpha-thrombin to RPRP, plasmin also increased the number of GMP-140 molecules expressed/platelet without enhancing alpha-thrombin binding to the platelets. Additionally, plasmin accelerated prothrombin activation when it was added to washed platelets resuspended in factor V depleted plasma simultaneously with 10 mM CaCl2, 10 nM alpha-thrombin for 10 s (to activate platelets and platelet factor V), followed by 4 microM hirudin and 1 nM factor Xa. Thus, plasmin potentiates the platelet release reaction in response to alpha-thrombin (probably by increasing the availability of factor V on the platelets) to enhance prothrombin activation in RPRP. These actions of plasmin may contribute to the increased platelet activation and thrombotic side-effects that can occur after thrombolytic therapy.


Pefloxacin penetration into human necrotic pancreatic tissue.
Antibiotic prophylaxis may be useful in acute necrotising pancreatis, a disease associated with a considerable incidence of infectious complications. The aim of this study was to assess pefloxacin penetration into necrotic pancreatic tissue during human necrotic pancreatitis. Ten patients (mean age 53.2 +/- 17.4 years) with severe acute pancreatitis (mean Ranson score 4.3) were studied. Pefloxacin was administered at a dose of 400 mg bd every 12 h by i.v. infusion (bolus, 15 min). Intraoperative samples of necrotic pancreatic tissue and blood were collected simultaneously 1, 2, 4.5, 6, 8.5 or 10 h after the last pefloxacin administration in patients treated for 1, 3, 4, 7, 8, 17 or 20 days. Drug concentrations were determined by the microbiological agar-well diffusion method (Escherichia coli Kp 05124 as test micro-organism in Isosensitest Agar). Levels in serum ranged from 2.0 to 9.0 mg/L (at 2 and 6 h, respectively), in necrotic pancreatic tissue from 2.0 to 29.0 micrograms/g depending on different sampling time. Maximum tissue peak concentrations appeared between 4 and 6 h. The necrotic pancreatic tissue/serum concentration ratio ranged from 0.9 to 5.1, values depending on tissue sample collection. Therapeutic concentrations (20.6 micrograms/g) above the MIC of potentially pathogenic enteric microorganisms were still present in necrotic pancreatic tissue 10 h after the last drug administration. Pefloxacin appeared to concentrate in necrotic pancreatic tissue, without appreciable accumulation after multiple-dose administration. The pefloxacin concentrations in necrotic pancreatic tissue showed high variability, depending on the degree of necrosis, inflammation and sample vascularization. Our results provided evidence of good, prompt penetration of pefloxacin into necrotic pancreatic tissue. Pefloxacin seems to exhibit favourable pharmacokinetic and pharmacodynamic properties for pancreatic infections.


Long-term stability of maxillary anterior alignment in non-extraction cases.
The purpose of this retrospective study was to evaluate long-term stability of maxillary incisors alignment in cases submitted to non-extraction orthodontic treatment. The sample comprised 23 patients (13 female; 10 male) at a mean initial age of 13.36 years (SD = 1.81 years), treated with fixed appliances. Dental cast measurements were obtained at three different time points (T1 - pretreatment, T2 - posttreatment and T3 - long-term posttreatment). Variables assessed in maxillary arch were Little Irregularity Index, intercanine, interpremolar and intermolar widths, arch length and perimeter. The statistical analysis was performed by one-way ANOVA and Tukey tests when necessary. Pearson' correlation coefficients were used to investigate possible associations between the evaluated variables. There was no significant change in most arch dimension measurements during and after treatment, however, during the long-term posttreatment period, it was observed a significant maxillary incisors crowding relapse. The maxillary incisors irregularity increased significantly (1.52 mm) during long-term posttreatment. None of the studied clinical factors demonstrated to be predictive of the maxillary crowding relapse.


The periaxonal space of crayfish giant axons.
The influence of the glial cell layer on effective external ion concentrations has been studied in crayfish giant axons. Excess K ions accumulate in the periaxonal space during outward K+ current flow, but at a rate far below that expected from the total ionic flux and the measured thickness of the space. At the conclusion of outward current flow, the external K+ concentration returns to normal in an exponential fashion, with a time constant of approximately 2 ms. This process is about 25 times faster than is the case in squid axons. K+ repolarization (tail) currents are generally biphasic at potentials below about -40 mV and pass through a maximum before approaching a final asymptotic level. The initial rapid phase may in part reflect depletion of excess K+. After block of inactivation and reversal of the Na+ concentration gradient, we could demonstrate accumulation and washout of excess Na ions in the periaxonal space. Characteristics of these processes appeared similar to those of K+. Crayfish glial cell ultrastructure has been examined both in thin sections and after freeze fracture. Layers of connective tissue and extracellular fluid alternate with thin layers of glial cytoplasm. A membranous tubular lattice, spanning the innermost glial layers, may provide a pathway allowing rapid diffusion of excess ions from the axon surface.


Towards a post-modern medicine.
Although there is general agreement that our current approach to health and healing is undergoing substantial change, there has been a lack of critical discussion regarding the extent, character and direction of change. It is important for us to articulate the values that we would like expressed in a reconfigured approach to health and healing, and to ensure that our efforts to initiate change are aligned with these values. An overview of Western history suggests that the post-modern world-view is characterized by four essential values: multidimensional realism, intentionality, holism and personal authenticity. Our current efforts to change the direction of health care have failed to produce fundamental change. This has resulted from the compelling influence of existing perspectives and the promotion of parochial, professional and institutional interests. To assure fundamental change, articulated values that are consistent with a post-modern world must be articulated, fostered and embedded into innovative programmes.


Continuous-flow centrifugation hemapheresis in the horse.
In a continuous-flow centrifugation apheresis technique adapted for blood-component separation and collection in horses, hydroxyethyl starch was not required for erythrocyte sedimentation. The efficacy and separation characteristics of whole blood from 10 horses were evaluated at various gravitational forces (700 to 1,500 rpm), using a constant withdrawal rate (100 ml/min). Maximum leukocyte collection occurred at 700 rpm (P less than 0.01), and optimal neutrophil collection occurred at 700 to 750 rpm (P less than 0.01). Although neutrophil counts decreased and lymphocyte counts remained constant at higher rpm settings, an optimal rpm range could not be determined for lymphocyte collection. Peak platelet collection occurred at 1,500 rpm. The response of whole blood from 5 horses was evaluated at lower (500 to 700 rpm) and higher (1,400 to 2,200 rpm) centrifuge speeds. A constant whole blood withdrawal rate of 100 ml/min and a leukocyte collection rate of 3 ml/min were maintained. The optimal rpm settings were determined and compared with values obtained from the 10 horses. Significant differences (P less than 0.01) did not exist between maximum numbers of leukocytes collected at low and high speeds. There was a significant (P less than 0.01) decrease in neutrophils and a concomitant increase in the numbers of lymphocytes recovered at the higher rpm. Platelet yields increased as rpm was increased to 2,000.


Weissella cibaria MG5285 and Lactobacillus reuteri MG5149 attenuated fat accumulation in adipose and hepatic steatosis in high-fat diet-induced C57BL/6J obese mice.
Excessive consumption of dietary fat is closely related to obesity, diabetes, insulin resistance, cardiovascular disease, hypertension, and non-alcoholic fatty liver disease. Recently, probiotics have been highly proposed as biotherapeutic to treat and prevent diseases. Previously, there are studies that demonstrated the beneficial effects of probiotics against metabolic disorders, including obesity and diabetes. We investigated the anti-obesity effect and mechanism of action of four human-derived lactic acid bacterial (LAB) strains (Lacticaseibacillus rhamnosus MG4502, Lactobacillus gasseri MG4524, Limosilactobacillus reuteri MG5149, and Weissella cibaria MG5285) in high-fat diet (HFD)-induced obese mice. Obesity was induced in mice over 8 weeks, with a 60% HFD. The four human-derived LAB strains (2 × 108 CFU/mouse) were orally administered to male C57BL/6J mice once daily for 8 weeks. Body weight, liver and adipose tissue (AT) weights, glucose tolerance, and serum biochemistry profiles were determined. After collecting the tissues, histopathological and Western blot analyses were conducted. Administration of these LAB strains resulted in decreased body weight, liver and AT weights, and glucose tolerance. Serum biochemistry profiles, including triglyceride (TG), total cholesterol, low-density lipoprotein cholesterol, and leptin, pro-inflammatory cytokines, improved. Hepatic steatosis and TG levels in liver tissue were significantly reduced. In addition, the size of adipocytes in epididymal tissue was significantly reduced. In epididymal tissues, Limosilactobacillus reuteri MG5149 and Weissella cibaria MG5285 groups showed a significantly reduced expression of lipogenic proteins, including peroxisome proliferator-activated receptor γ, CCAAT/enhancer-binding protein α, fatty acid synthase (FAS), and adipocyte-protein 2. In addition, sterol regulatory element-binding protein 1-c and its downstream protein FAS in the liver tissue were significantly decreased. These strains attenuated fat accumulation in the liver and AT by upregulating the phosphorylation of AMP-activated protein kinase and acetyl-CoA carboxylase in HFD-fed mice. We suggest that L. reuteri MG5149 and W. cibaria MG5285 could be used as potential probiotic candidates to prevent obesity.


Time spent on work-related activities, social activities and time pressure as intermediary determinants of health disparities among elderly women and men in 5 European countries: a structural equation model.
Psychosocial factors shape the health of older adults through complex inter-relating pathways. Besides socioeconomic factors, time use activities may explain gender inequality in self-reported health. This study investigated the role of work-related and social time use activities as determinants of health in old age. Specifically, we analysed whether the impact of stress in terms of time pressure on health mediated the relationship between work-related time use activities (i.e. housework and paid work) on self-reported health. We applied structural equation models and a maximum-likelihood function to estimate the direct and indirect effects of psychosocial factors on health using pooled data from the Multinational Time Use Study on 11,168 men and 14,295 women aged 65+ from Italy, Spain, UK, France and the Netherlands. The fit indices for the conceptual model indicated an acceptable fit for both men and women. The results showed that socioeconomic status (SES), demographic factors, stress and work-related time use activities after retirement had a significant direct influence on self-reported health among the elderly, but the magnitude of the effects varied by gender. Social activities had a positive impact on self-reported health but had no significant impact on stress among older men and women. The indirect standardized effects of work-related activities on self-reported health was statistically significant for housework (β = - 0.006; P < 0.001 among men and β = - 0.008; P < 0.001 among women) and paid work (β = 0.012; P < 0.01 among men and β = 0.000; P > 0.05 among women), which implied that the paths from paid work and housework on self-reported health via stress (mediator) was very weak because their indirect effects were close to zero. Our findings suggest that although stress in terms of time pressure has a direct negative effect on health, it does not indirectly influence the positive effects of work-related time use activities on self-reported health among elderly men and women. The results support the time availability hypothesis that the elderly may not have the same time pressure as younger adults after retirement.


CytoBayesJ: software tools for Bayesian analysis of cytogenetic radiation dosimetry data.
A number of authors have suggested that a Bayesian approach may be most appropriate for analysis of cytogenetic radiation dosimetry data. In the Bayesian framework, probability of an event is described in terms of previous expectations and uncertainty. Previously existing, or prior, information is used in combination with experimental results to infer probabilities or the likelihood that a hypothesis is true. It has been shown that the Bayesian approach increases both the accuracy and quality assurance of radiation dose estimates. New software entitled CytoBayesJ has been developed with the aim of bringing Bayesian analysis to cytogenetic biodosimetry laboratory practice. CytoBayesJ takes a number of Bayesian or 'Bayesian like' methods that have been proposed in the literature and presents them to the user in the form of simple user-friendly tools, including testing for the most appropriate model for distribution of chromosome aberrations and calculations of posterior probability distributions. The individual tools are described in detail and relevant examples of the use of the methods and the corresponding CytoBayesJ software tools are given. In this way, the suitability of the Bayesian approach to biological radiation dosimetry is highlighted and its wider application encouraged by providing a user-friendly software interface and manual in English and Russian.


Display of major risk categories for android apps.
Communicating cybersecurity risks to mobile-device users is essential. However, existing means of conveying the risks through detailed permission lists are ineffective. Risk indexes that convey overall risk are effective at influencing app-selection decisions, but many users want more information. We examined how users assess the risks associated with downloading applications on the Android platform by comparing various graphical formats of intermediate-level risk displays containing three risk categories: personal privacy; monetary loss; device stability. Bar-graph and table formats were compared, as were vertical and horizontal displays. Participants performed app risk-rating (Experiment 1) and app-selection (Experiments 2 and 3) tasks for hypothetical apps with risk scores on each of the categories. They also specified which risk category was of most concern to them. Increased risk scores in each category led to higher rated risk and lower app-selection rate which matched with self-reported risk concerns. Bar-graphs were more time-efficient and yielded higher risk-ratings than tables, although the two formats did not differ in the app-selection task. Moreover, horizontal bar-graphs yielded faster responses than vertical bar-graphs. The results indicate that the intermediate-level risk display was effective in conveying risk-category information, especially with horizontal bar-graphs, and personalized design of this display based on users' risk concerns is potentially useful. (PsycINFO Database Record


Immunocytochemical localization of intrinsic factor--cobalamin bound to the guinea pig ileum in vivo.
The absorption of dietary cobalamin requires that a complex of intrinsic factor-cobalamin bind to specific receptors in the distal small intestine. The cellular location of these binding sites is unknown. To localize the intrinsic factor-cobalamin binding site, homogeneously purified hog intrinsic factor (7 pmol) and cobalamin (7 pmol) were instilled into ileal loops (n = 10) in anesthetized guinea pigs. Tissue controls for binding specificity were intrinsic factor-cobalamin in colon (n = 3) and jejunal (n = 3) loops, as well as intrinsic factor alone in ileal loops (n = 3). Intrinsic factor-[57Co]cobalamin was instilled in three ileal loops maintained for 90 min, and the radioactivity in the liver and kidneys was measured. Portions of each loop were washed in Krebs-Ringer buffer + Ca++, pH 7.4, or in either 1 mM Na2 ethylenediaminetetraacetic acid or in pH 4.5 acetate buffer. All tissue was processed for ultrastructural immunocytochemistry using a monospecific antisera to hog intrinsic factor as part of a well-characterized indirect immunoperoxidase method. Essentially all immunoreactive intrinsic factor-cobalamin was associated with the microvillous pits on mature ileal absorptive cells. Intrinsic factor binding was not present in colon or jejunum, or in the ileal loops instilled with intrinsic factor alone. Ethylenediaminetetraacetic acid and acid pH decreased intrinsic factor-cobalamin binding but caused severe disruption of the microvillous architecture. Absorption of [57Co]cobalamin was demonstrated, but intrinsic factor was not identified within enterocytes. The localization of specific intrinsic factor-cobalamin binding sites to the microvillous pit suggests that the intrinsic factor-cobalamin receptor may maintain a relatively fixed position within the surface membrane, and although alternative explanations are possible, our findings support the hypothesis that intrinsic factor is not internalized into the enterocyte during cobalamin absorption.


ZOLA-3D allows flexible 3D localization microscopy over an adjustable axial range.
Single molecule localization microscopy can generate 3D super-resolution images without scanning by leveraging the axial variations of normal or engineered point spread functions (PSF). Successful implementation of these approaches for extended axial ranges remains, however, challenging. We present Zernike Optimized Localization Approach in 3D (ZOLA-3D), an easy-to-use computational and optical solution that achieves optimal resolution over a tunable axial range. We use ZOLA-3D to demonstrate 3D super-resolution imaging of mitochondria, nuclear pores and microtubules in entire nuclei or cells up to ~5 μm deep.


An investigation of the protein components of amyloid using immunoperoxidase and permanganate methods on tissue sections.
Amyloid deposits in tissue from 8 patients with generalized primary amyloidosis, 11 patients with generalized secondary amyloidosis, 11 nasopharyngeal carcinomas, 11 basal cell carcinomas, 4 islet cell tumours, 4 medullary carcinomas of the thyroid and 9 cases of lichen amyloidosis were studied using the indirect immunoperoxidase and peroxidase-antiperoxidase methods with specific antisera against Amyloid A (AA) protein and human immunoglobulin lambda and kappa light chains. The permanganate method of Wright was also applied to tissue sections. Positive staining for AA protein was observed only in secondary amyloidosis. There was excellent correlation between AA positivity and permanganate sensitivity. Positivity for immunoglobulin light chains was not observed in secondary amyloidosis but was noted in 5 (63%) cases of primary amyloidosis and 18-27% of intratumour amyloidosis. Lichen amyloidosis did not stain for AA protein or light chains. It is shown that assessment of the permanganate reaction and AA positivity of amyloid deposits can reliably differentiate secondary from primary amyloidosis and may contribute significantly to selection of patients for appropriate therapy.


Ionizing radiation abrogates the pro-tumorigenic capacity of cancer-associated fibroblasts co-implanted in xenografts.
Cancer-associated fibroblasts (CAFs) are abundantly present in solid tumors and affect tumorigenesis and therapeutic responses. In the context of clinical radiotherapy, the impact of irradiated CAFs to treatment outcomes is largely unexplored. Aiming at improving radiotherapy efficacy, we have here explored the effect of radiation on the inherent pro-tumorigenic capacity of CAFs in animals. Ionizing radiation was delivered to cultured CAFs as single-high or fractionated doses. Tumor development was compared in mice receiving A549 lung tumor cells admixed with irradiated or control CAFs. Biological mechanisms behind tumor growth regulation were investigated by quantitative histology and immunohistochemistry. Viability assessments confirmed that irradiated CAFs are fully functional prior to implantation. However, the enhanced tumorigenic effect observed in tumors co-implanted with control CAFs was abrogated in tumors established with irradiated CAFs. Experiments to ascertain fate of implanted fibroblasts showed that exogenously administered CAFs reside at the implantation site for few days, suggesting that tumor growth regulation from admixed CAFs take place during initial tumor formation. Our work demonstrate that irradiated CAFs lose their pro-tumorigenic potential in vivo, affecting angiogenesis and tumor engraftment. This finding propose a previously unknown advantageous effect induced by radiotherapy, adding to the direct cytotoxic effects on transformed epithelial cells.


A pH-regulated dimeric bouquet in the structure of von Willebrand factor.
At the acidic pH of the trans-Golgi and Weibel-Palade bodies (WPBs), but not at the alkaline pH of secretion, the C-terminal ∼1350 residues of von Willebrand factor (VWF) zip up into an elongated, dimeric bouquet. Six small domains visualized here for the first time between the D4 and cystine-knot domains form a stem. The A2, A3, and D4 domains form a raceme with three pairs of opposed, large, flower-like domains. N-terminal VWF domains mediate helical tubule formation in WPBs and template N-terminal disulphide linkage between VWF dimers, to form ultralong VWF concatamers. The dimensions we measure in VWF at pH 6.2 and 7.4, and the distance between tubules in nascent WPB, suggest that dimeric bouquets are essential for correct VWF dimer incorporation into growing tubules and to prevent crosslinking between neighbouring tubules. Further insights into the structure of the domains and flexible segments in VWF provide an overall view of VWF structure important for understanding both the biogenesis of ultralong concatamers at acidic pH and flow-regulated changes in concatamer conformation in plasma at alkaline pH that trigger hemostasis.


Active space of pheromone plume and its relationship to effective attraction radius in applied models.
The release rate of a semiochemical lure that attracts flying insects has a specific effective attraction radius (EAR) that corresponds to the lure's orientation response strength. EAR is defined as the radius of a passive sphere that intercepts the same number of insects as a semiochemical-baited trap. It is estimated by calculating the ratio of trap catches in the field in baited and unbaited traps and the interception area of the unbaited trap. EAR serves as a standardized method for comparing the attractive strengths of lures that is independent of population density. In two-dimensional encounter rate models that are used to describe insect mass trapping and mating disruption, a circular EAR (EAR(c)) describes a key parameter that affects catch or influence by pheromone in the models. However, the spherical EAR, as measured in the field, should be transformed to an EAR(c) for appropriate predictions in such models. The EAR(c) is calculated as (pi/2EAR(2))/F (L), where F (L) is the effective thickness of the flight layer where the insect searches. F (L) was estimated from catches of insects (42 species in the orders Coleoptera, Lepidoptera, Diptera, Hemiptera, and Thysanoptera) on traps at various heights as reported in the literature. The EAR(c) was proposed further as a simple but equivalent alternative to simulations of highly complex active-space plumes with variable response surfaces that have proven exceedingly difficult to quantify in nature. This hypothesis was explored in simulations where flying insects, represented as coordinate points, moved about in a correlated random walk in an area that contained a pheromone plume, represented as a sector of active space composed of a capture probability surface of variable complexity. In this plume model, catch was monitored at a constant density of flying insects and then compared to simulations in which a circular EAR(c) was enlarged until an equivalent rate was caught. This demonstrated that there is a circular EAR(c), where all insects that enter are caught, which corresponds in catch effect to any plume. Thus, the EAR(c), based on the field-observed EAR, can be used in encounter rate models to develop effective control programs based on mass trapping and/or mating disruption.


Physicochemical stability of voriconazole in elastomeric devices.
Voriconazole is the drug of choice for invasive aspergillosis (IA), a leading cause of mortality and morbidity in immunocompromised patients. Prolong intravenous administration of voriconazole is often needed in such patients due to high incidence of oral mucositis and unreliable bioavailability of oral dosage form. Administration of voriconazole through elastomeric pump may facilitate early hospital discharge of clinically stable immunocompromised patients needing prolonged intravenous treatment. Therefore, we investigated the physicochemical stability of voriconazole in one of the commonly used elastomeric pumps at three different temperatures for various time points. A total of 18 elastomeric pumps were prepared and 6 containing 2 mg/mL of voriconazole (3 in 0.9% sodium chloride and 3 in 5% glucose) were stored at either 4°C for 96 hours, 25°C for 4 hours or at 35°C for 4 hours. An aliquot withdrawn immediately before storage (time 0) and at various time points was analysed for chemical stability using high-performance liquid chromatography and for physical stability using visual, pH and microscopic analyses. Voriconazole was stable for at least 96 hours, 4 hours and 4 hours at 4°C, 25°C and 35°C, respectively, when admixed with either 0.9% sodium chloride or 5% glucose. No evidence of particle formation, colour change or pH change was observed throughout the study period. These findings would allow early hospital discharge using elastomeric intravenous administration of voriconazole in patients in whom oral route of administration is not available.


Continuous versus intermittent beta-agonists in the treatment of acute asthma.
Patients with acute asthma treated in the emergency department are frequently treated with intermittent inhaled beta-agonists delivered by nebulisation. The use of continuous beta-agonist (CBA) via nebulisation in the emergency setting may offer additional benefits in acute asthma. To determine the efficacy (e.g., reductions in admission, improvement in pulmonary functions) and risks (e.g., adverse events, effects on vital signs) of continuous versus intermittent inhaled beta-agonists for the treatment of patients with acute asthma managed in the emergency department. Randomised controlled trials were identified from the Cochrane Airways Review Group "Asthma and WHEEZ*" Register which is a compilation of systematic searches of CINAHL, EMBASE, MEDLINE and CENTRAL and hand searching of 20 respiratory journals. In addition, primary authors and content experts were contacted to identify eligible studies. Bibliographies from included studies, known reviews and texts were also searched. The search is considered updated to June 2003. Only randomised controlled trials (RCTs) were eligible for inclusion. Studies were included if patients presented with acute asthma and were treated with either continuous or intermittent inhaled beta-agonists early in the ED treatment. "Continuous" nebulisation was defined as truly continuous aerosol delivery of beta-agonist medication (e.g., using a commercially available large-volume nebuliser, or a small-volume nebuliser with infusion pump) or sufficiently frequent nebulisations that medication delivery was effectively continuous (i.e., 1 nebulisation every 15 minutes or > 4 nebulisations per hour). Studies also needed to report either pulmonary function or admission results. Two reviewers independently selected potentially relevant articles and two additional reviewers independently selected articles for inclusion. Methodological quality was independently assessed by two reviewers. Data were extracted independently by two reviewers if the authors were unable to verify the validity of information. Missing data were obtained from authors or calculated from other data presented in the paper. The data were analysed using the Cochrane Review Manager (Version 4.1). Relative risks (RR), weighted mean differences (WMD) and standardized mean differences (SMD) are reported with corresponding 95% confidence intervals (CI); both peak expiratory flow rates (PEFR) and forced expiratory volume in one second (FEV-1) data are reported. 165 trials were reviewed and eight were included; a total of 461 patients have been studied (229 with CBA; 232 with intermittent beta-agonists). Overall, admission to hospital was reduced with CBA compared to intermittent beta-agonists (RR: 0.68; 95% CI: 0.5 to 0.9); patients with severe airway obstruction at presentation appeared to benefit most from this intervention (RR: 0.64; 95% CI: 0.5 to 0.9). Patients receiving CBA demonstrated small but statistically significant improvements in pulmonary function tests when all studies were pooled. Patients receiving CBA had greater improvements in % predicted FEV-1 (SMD: 0.3; 95% CI: 0.03 to 0.5) and PEFR (SMD: 0.33; 95% CI: 0.1 to 0.5); this effect was observed by 2-3 hours. Continuous treatment was generally well tolerated, with no clinically important differences observed in pulse rate (WMD: -2.87; 95% CI: -6.0 to 0.3) or blood pressure (WMD: -1.75; 95% CI: -5.6 to 2.1) between the treatment groups. Tremor was equally common in both groups (OR: 0.81; 95% CI: 0.5 to 1.3) and potassium concentration was unchanged (WMD: 0.02; 95% CI: -0.2 to 0.2). Current evidence supports the use of CBA in patients with severe acute asthma who present to the emergency department to increase their pulmonary functions and reduce hospitalisation. Moreover, CBA treatment appears to be safe and well tolerated in patients who receive it.


Earliest known crown-group salamanders.
Salamanders are a model system for studying the rates and patterns of the evolution of new anatomical structures. Recent discoveries of abundant Late Jurassic and Early Cretaceous salamanders are helping to address these issues. Here we report the discovery of well-preserved Middle Jurassic salamanders from China, which constitutes the earliest known record of crown-group urodeles (living salamanders and their closest relatives). The new specimens are from the volcanic deposits of the Jiulongshan Formation (Bathonian), Inner Mongolia, China, and represent basal members of the Cryptobranchidae, a family that includes the endangered Asian giant salamander (Andrias) and the North American hellbender (Cryptobranchus). These fossils document a Mesozoic record of the Cryptobranchidae, predating the previous record of the group by some 100 million years. This discovery provides evidence to support the hypothesis that the divergence of the Cryptobranchidae from the Hynobiidae had taken place in Asia before the Middle Jurassic period.


Pro-inflammatory and antiviral cytokine expression in vaccinated and unvaccinated horses exposed to equine influenza virus.
Most studies of the cytokine response to influenza virus infection have been carried out in human, porcine and murine models, however the data available on equine cytokines is limited. An experimental challenge study was undertaken in unvaccinated naïve horses and horses vaccinated with a commercial inactivated influenza vaccine. The humoral antibody response to vaccination and virus challenge was measured by single radial haemolysis (SRH) assay and clinical signs of influenza and viral shedding were monitored post-challenge. Levels of three equine pro-inflammatory cytokines interleukin (IL)-1beta, IL-6 and tumor necrosis factor (TNF)-alpha and the antiviral cytokine interferon (IFN)-alpha were examined by quantitative RT-PCR of mRNA. Vaccination provided significant clinical and virological protection and resulted in a significant reduction of IFN-alpha and IL-6 expression on day 2 post-challenge. The patterns of cytokine expression observed in control animals suffering from influenza after challenge are comparable to those reported in studies of other species.


A 10-year longitudinal study of dental students' emotional intelligence and the impact of COVID-19.
Emotional intelligence (EI) supports the clinical and social competencies of a practicing dentist. Reuven Bar-On's EI model is an array of inter-related emotional and social competencies, skills, and behaviors, which consist of five key domains: Self-Perception, Self-Expression, Interpersonal, Decision Making, and Stress Management, and associated with the domains are 15 emotional quotient (EQ) subskills. This study measured the impact of COVID-19 on dental students' EI by comparing measures pre-COVID-19 and during COVID-19 matriculation. This retrospective longitudinal study measured EI with the EQ-i 2.0 for higher education. Dental students completed an EQ-i 2.0 assessment (Attempt) at the beginning of matriculation, at the mid-point, and prior to graduation. Ten groups were included, of which the first three completed matriculation prior to the pandemic and the remaining seven matriculated during timeframes intersecting at different times during the pandemic. A paired t-test dependent sample of means (p ≤ 0.05) compared EQ scores for each attempt for all groups. The study compared means for three EQ attempts with the t-test independent sample of means (p ≤ 0.05) for cohorts matriculating pre-COVID-19 and during COVID-19. The pre-COVID-19 groups showed significant increases in EQ with each subsequent attempt. COVID-19-impacted groups demonstrated significant increase in Stress Tolerance and significant decreases, most notably in the domains of Interpersonal and Self-Perception, and subscales of Optimism and Happiness. COVID-19-related stressors impacted dental students' EI as multiple EI areas declined significantly. Dental educators should minimize organizational stressors and support EI during years 2 and 3 of matriculation.


Effect of Type 2 Diabetes Mellitus on the Hypoxia-Inducible Factor 1-Alpha Expression. Is There a Relationship with the Clock Genes?
Limited reports exist on the relationships between regulation of oxygen homeostasis and circadian clock genes in type 2 diabetes. We examined whether the expression of Hypoxia-Inducible Factor-1α (HIF-1α) and HIF-2α relates to changes in the expression of clock genes (Period homolog proteins (PER)1, PER2, PER3, Retinoid-related orphan receptor alpha (RORA), Aryl hydrocarbon receptor nuclear translocator-like protein 1 (ARNTL), Circadian locomotor output cycles kaput (CLOCK), and Cryptochrome proteins (CRY) 1 and CRY2) in patients with type 2 diabetes. A total of 129 subjects were evaluated in this cross-sectional study (48% with diabetes). The gene expression was measured by polymerase chain reaction. The lactate and pyruvate levels were used as surrogate of the hypoxia induced anaerobic glycolysis activity. Patients with diabetes showed an increased plasma concentration of both lactate (2102.1 ± 688.2 vs. 1730.4 ± 694.4 uM/L, p = 0.013) and pyruvate (61.9 ± 25.6 vs. 50.3 ± 23.1 uM/L, p = 0.026) in comparison to controls. However, this finding was accompanied by a blunted HIF-1α expression (1.1 (0.2 to 5.0) vs. 1.7 (0.4 to 9.2) arbitrary units (AU), p ≤ 0.001). Patients with diabetes also showed a significant reduction of all assessed clock genes' expression. Univariate analysis showed that HIF-1α and almost all clock genes were significantly and negatively correlated with HbA1c concentration. In addition, positive correlations between HIF-1α and the clock genes were observed. The stepwise multivariate regression analysis showed that HbA1c and clock genes independently predicted the expression of HIF-1α. Type 2 diabetes modifies the expression of HIF-1α and clock genes, which correlates with the degree of metabolic control.