Alpha-particle-induced p53 protein expression in a rat lung epithelial cell strain.
Other investigators have shown that both sparsely ionizing and UV radiation cause cell cycle arrest that is associated with increased expression of wild-type p53 protein. The effect of exposure to alpha-particles from 238Pu on the induction of the p53 protein has now been examined in cultured lung epithelial cells derived from male F344 rats. The number of cells having increased levels of p53 protein was determined by flow cytometry after the cells had been stained with a monoclonal antibody to p53. alpha-Particle irradiation caused a dose-dependent increase in p53 protein levels detectable at doses as low as 0.6 cGy, with no evidence of a threshold. An increase in p53 protein also occurred in X-irradiated cells. However, no increase was seen in cells exposed to less than 10 cGy of X-rays, indicating the existence of a relatively higher DNA damage threshold for sparsely ionizing radiation. In addition, more cells exposed to low doses of alpha radiation had increased p53 protein levels than would be predicted based on the number of nuclei expected to be traversed by an alpha-particle, suggesting that alpha-particles cause genetic damage by mechanisms in addition to direct interactions with DNA.


Drug-induced lung disease: 1990 review.
Numerous drug-induced pulmonary manifestations have been reported but studies of their pathogenic mechanisms are still rare. These mechanisms should, however, be precisely determined in order to identify subjects at risk and to prevent some of these complications by the proper use of certain drugs in more appropriate conditions. The possibility of an iatrogenic manifestation should always be considered in patients developing pulmonary symptoms. Data from biological investigations, although not specific, contribute to the understanding of lung injury mechanisms.


Cloning, preparation and characterization of biologically active recombinant caprine placental lactogen.
Caprine placental lactogen (cPL) cDNA was cloned by reverse transcription (RT)-PCR from total RNA of goat placenta. The PCR product encoding for the mature protein was gel purified, ligated to pGEM-T and finally subcloned into a pET8c prokaryotic expression vector. E. coli cells (BL-21) transformed with this vector overexpressed large amounts of cPL upon induction with Isopropyl-1-thio-beta-D-galactopyranoside. The expressed protein, found in the inclusion bodies, was refolded and purified to homogeneity on Q-Sepharose and SP-Sepharose columns, yielding two electrophoretically pure fractions (cPL-Q and cPL-S), composed of over 98% of monomeric protein of the expected molecular mass of approximately 23 kDa. Binding of cPL to the extracellular domain (ECD) of prolactin receptors (PRLR) from rat (r), rabbit (rb), and bovine (b), growth hormone receptors (GHR) from human (h) and rabbit, and binding to rabbit mammary gland membranes revealed similar binding profiles for cPL-Q, cPL-S and ovine (o)PL. Caprine PL was capable of forming 1:2 complexes with hGHR-ECD, rbGHR-ECD, rPRLR-ECD and rbPRLR-ECD whereas with bPRLR-ECD only a 1:1 complex was detected. The biological activity of both cPL fractions resulting from proper renaturation was further evidenced by their ability to stimulate proliferation of Nb2 cells, FDC-P1 cells transfected with rabbit or human GHRs and by stimulation of beta-casein synthesis in rabbit and ovine mammary gland acini cultures.


Ecotypic differentiation and phenotypic plasticity in reproductive traits of Armadillidium vulgare (Isopoda: Oniscidea).
Armadillidium vulgare differed in growth and survivorship on two field sites. Growth rates were higher at a site with consistently higher quality food than at the other site where less high-quality food was produced and which was less predictably accessible. Survivorship was higher at the second site where temperature fluctuations were consistently smaller. Individuals from the two populations were kept for 6 months under the same food and temperature conditions and patterns of resource allocation to reproductive traits analysed. Members of the population from the site with good growth conditions had significantly higher reproductive allocation, by 13.5%, and larger broods, by 9.1%, than those from the site with poor growth conditions. Contrary to theoretical predictions, they also had significantly larger offspring, by 7.5% dry mass. Larger offspring survived better than small ones. This differential survivorship, by 20% for a 3.4% difference in live mass, was much more pronounced under conditions of moisture stress and under fluctuating temperature regimes. Larger offspring would therefore be at a selective advantage on the site with more severe temperature fluctuations. Phenotypic plasticity in reproductive traits in response to experimental changes in food quality, temperature and crowding were monitored. Reproductive allocation was increased by 20.8% under conditions of higher food quality, by 14.7% at higher temperatures, and by 12.5% under less crowded conditions. Brood size, but not offspring dry mass, increased when food quality increased. When crowding increased by 25.0%, the size of broods remained the same but the dry mass of individual offspring decreased by 11.2%. Members of the population from the site with more variable access to high-quality food showed more plasticity in reproductive traits in response to changes in food supply than members of the population from the site with the more predictable food supply. Members of the population from the site with more stable temperatures showed less plasticity to temperature changes than members of the population from the site with greater temperature fluctuations. It is concluded that the observed microevolutionary processes and phenotypic plasticity have adaptative value as responses to spatial and temporal heterogeneity in environmental conditions.


Natural Correction and Adaptation of a Severely Deformed Sacral Dome in an Adolescent with Isthmic Spondylolisthesis: A Case Report.
A spinal deformity with wedging of L5 and rounding of the surface of the sacral dome is very common in pediatric patients with spondylolisthesis at the L5 vertebral level, and it has been well documented that severe spondylolisthesis is a good indication for spinal arthrodesis. We report the natural correction and adaptation of a severely deformed sacral dome with slippage (40.8%) in a pediatric patient with nonoperative treatment. Vertebral deformity in children can be reversible. Orthopaedic surgeons should be aware that pediatric patients with a severe deformity of the sacral dome and vertebral slippage can be managed nonoperatively.


Measured and predicted bone mineral content in healthy boys and girls aged 6-18 years: adjustment for body size and puberty.
Dual-energy X-ray absorptiometry (DEXA) is a rapid and precise technique for the assessment of bone mineralization in children. Interpretation of the results in growing children is complex as results are influenced by age, body size (height and weight) and puberty. Conventionally, bone mineral data derived from DEXA have been presented as an areal density [BMD; bone mineral content (BMC, g)/projected bone area (BA, cm2)], yet this fails to account for changes in BMC that result from changes in age, body size or pubertal development. Measurement of BMC and BA of the whole body, lumbar spine and left hip were made in 58 healthy boys and girls using DEXA. The relationship between BMC and BA was curvilinear, with the best fit being that of a power model (BMD = BMC/BAlambda, where lambda is the exponent to which BA is raised in order to remove its influence on BMC). The value of lambda changed when measures of body size and puberty were taken into account (e.g. for lumbar spine from 1.66 to 1.49). Predictive formulae for BMC were produced using regression analysis and based on the variables of age, body size and pubertal development. This provides a method for interpreting the measured BMC which is independent of such variables and a constant reference range for children aged 6-18 y.


Brain Atrophy and the Risk of Futile Endovascular Reperfusion in Acute Ischemic Stroke.
Background and Purpose- We aimed to evaluate the impact of brain atrophy on long-term clinical outcome in patients with acute ischemic stroke treated with endovascular therapy, and more specifically, to test whether there are interactions between the degree of atrophy and infarct volume, and between atrophy and age, in determining the risk of futile reperfusion. Methods- We studied consecutive patients with acute ischemic stroke with proximal anterior circulation intracranial arterial occlusions treated with endovascular therapy achieving successful arterial recanalization. Brain atrophy was evaluated on baseline computed tomography with the global cortical atrophy scale, and Evans index was calculated to assess subcortical atrophy. Infarct volume was assessed on control computed tomography at 24 hours using the formula for irregular volumes (A×B×C/2). Main outcome variable was futile recanalization, defined by functional dependence (modified Rankin Scale score >2) at 3 months. The predefined interactions of atrophy with age and infarct volume were studied in regression models. Results- From 361 consecutive patients with anterior circulation acute ischemic stroke treated with endovascular therapy, 295 met all inclusion criteria. Futile reperfusion was observed in 144 out of 295 (48.8%) patients. Cortical atrophy affecting parieto-occipital and temporal regions was associated with futile recanalization. Total global cortical atrophy score and Evans index were independently associated with futile recanalization in an adjusted logistic regression. Multivariable adjusted regression models disclosed significant interactions between global cortical atrophy score and infarct volume (odds ratio, 1.003 [95%CI, 1.002-1.004], P<0.001) and between global cortical atrophy score and age (odds ratio, 1.001 [95% CI, 1.001-1.002], P<0.001) in determining the risk of futile reperfusion. Conclusions- A higher degree of cortical and subcortical brain atrophy is associated with futile endovascular reperfusion in anterior circulation acute ischemic stroke. The impact of brain atrophy on insufficient clinical recovery after endovascular reperfusion appears to be independently amplified by age and by infarct volume.


Depletion of securin increases arsenite-induced chromosome instability and apoptosis via a p53-independent pathway.
Arsenic is a pathologic factor of cardiovascular diseases and cancers; nevertheless, it also acts as an anticancer agent effective on acute promyelocytic leukemia and multiple myeloma. Securin, a proposed proto-oncogene, regulates cell proliferation and tumorigenesis. However, roles of securin on the arsenic-induced cell cycle arrest and apoptosis remain unknown. In this study, the effects of sodium arsenite on the expression of securin in two tissue types of cell lines, the vascular endothelial and colorectal epithelial cells, were investigated. Arsenite (8-16 microM, 24 h) increased the cytotoxicity, apoptosis, and growth inhibition in both endothelial and epithelial cells. The levels of phospho-CDC2 (threonine-161), CDC2, and cyclin B1 proteins were decreased, and the G2/M fractions were increased by arsenite. Concomitantly, arsenite markedly diminished the securin protein expression and induced the abnormal sister chromatid separation. The depletion of securin proteins increased the induction of mitotic arrest, aberrant chromosome segregation, and apoptosis after arsenite treatment. p53, a tumor suppressor protein, balances the cell survival and apoptosis. Arsenite raised the levels of phospho-p53 (serine-15) and p53 (DO-1) proteins in both the securin-wild-type and -null cells. The p53-functional cells were more susceptible than the p53-mutational cells to arsenite on the cytotoxicity and apoptosis. Besides, arsenite decreased the levels of securin proteins to a similar degree in both the p53-functional and -mutational cells. Together, it is the first time to demonstrate that the inhibition of securin expression induced by arsenite increases the chromosomal instability and apoptosis via a p53-independent pathway.


'maskBAD'--a package to detect and remove Affymetrix probes with binding affinity differences.
Hybridization differences caused by target sequence differences can be a confounding factor in analyzing gene expression on microarrays, lead to false positives and reduce power to detect real expression differences. We prepared an R Bioconductor compatible package to detect, characterize and remove such probes in Affymetrix 3'IVT and exon-based arrays on the basis of correlation of signal intensities from probes within probe sets. Using completely mouse genomes we determined type 1 (false negatives) and type 2 (false positives) errors with high accuracy and we show that our method routinely outperforms previous methods. When detecting 76.2% of known SNP/indels in mouse expression data, we obtain at most 5.5% false positives. At the same level of false positives, best previous method detected 72.6%. We also show that probes with differing binding affinity both hinder differential expression detection and introduce artifacts in cancer-healthy tissue comparison. Detection and removal of such probes should be a routine step in Affymetrix data preprocessing. We prepared a user friendly R package, compatible with Bioconductor, that allows the filtering and improving of data from Affymetrix microarrays experiments.


A leading role for the immune system in the pathophysiology of preeclampsia.
Preeclampsia syndrome is characterized by inadequate placentation, because of deficient trophoblastic invasion of the uterine spiral arteries, leading to placental hypoxia, secretion of proinflammatory cytokines, the release of angiogenic and antiangiogenic factors and miRNAs. Although immune-system alterations are associated with the origin of preeclampsia, other factors, including proinflammatory cytokines, neutrophil activation, and endothelial dysfunction, are also related to the pathophysiology of this syndrome. The pathophysiology of preeclampsia may involve several factors, including persistent hypoxia at the placental level and the release of high amounts of STBMs. DAMP molecules released under hypoxic conditions and STBMs, which bind TLRs, may activate monocytes, DCs, NK cells, and neutrophils, promoting persistent inflammatory conditions in this syndrome. The development of hypertension in preeclamptic women is also associated with endothelial dysfunction, which may be mediated by various mechanisms, including neutrophil activation and NET formation. Furthermore, preeclamptic women have higher levels of nonclassic and intermediate monocytes and lower levels of lymphoid BDCA-2(+) DCs. The cytokines secreted by these cells may contribute to the inflammatory process and to changes in adaptive-immune system cells, which are also modulated in preeclampsia. The changes in T cell subsets that may be seen in preeclampsia include low Treg activity, a shift toward Th1 responses, and the presence of Th17 lymphocytes. B cells can participate in the pathophysiology of preeclampsia by producing autoantibodies against adrenoreceptors and autoantibodies that bind the AT1-R.


Complex nonunion of fractures of the femoral shaft treated by wave-plate osteosynthesis.
We have treated 42 consecutive complex ununited fractures of the femoral shaft by wave-plate osteosynthesis at five different medical centres. There were 13 with previous infection, 12 with segmental cortical defects, and 3 were pathological fractures. In 39 cases there had been previous internal fixation and 21 patients had had more than one earlier operation. Union was achieved in 41 patients at an average of six months, although three had required a second bone graft. Two patients had recurrence of infection and in one this resulted in the persistence of nonunion. There were no failures of the implant. All 41 patients with union are now fully weight-bearing, but four have a leg-length discrepancy, one has axial malalignment, and nine have residual stiffness of the knee. These results are surprisingly good, despite the complexity of the initial problem, and appear to confirm the biological and mechanical advantages of the wave plate over the conventional plate for such cases.


[Subcapsular hematoma of liver during pregnancy: report of 4 cases].
A Subcapsular hematoma of the liver is a rare but very serious complication of pregnancy. Diagnosis is confirmed at imaging (Ultrasound, CT, MRI) since clinical symptoms are not specific and biological signs have a delayed manifestation. The management of this complication depends on the integrity or rupture of Glisson's capsule. We report the cases of 4 patients and review of the literature.


Three staged minimally invasive esophagectomy with end-to-end esophago-gastric anastomosis for thoracic esophageal cancers: An experience from a Low Middle-Income Country.
Esophageal cancer is on a steady rise and carries significant mortality and morbidity. Depending upon the clinical stage at presentation, either chemotherapy, radiotherapy with or without surgical resection is the treatments in practice. Traditionally, open esophagectomy was performed but over time, the importance of minimally invasive esophagectomy has been established. In this study, we aimed to report our data of totally minimally invasive esophagectomies performed for thoracic esophageal cancers in last four years. A prospective cross-sectional study was conducted at the Department of Upper GI Surgery, Dow University of Health Sciences, Karachi. All diagnosed cases of esophageal carcinoma undergoing minimally invasive esophagectomy, from 2019 to 2022 were included in this study. Outcomes measured were operative time, intra operative complications, conversion rate to open, postoperative complications, number of lymph nodes harvested, margin clearance, in-hospital mortality and 90-days mortality. A total of 53 cases were included in the study, the most prevalent histological type was squamous cell carcinoma 42(79.2%) as compared to adenocarcinoma 8(15.1%). Most common tumor site was lower thoracic esophagus (30-38 cm) in 20 (56.6%) cases. Neo-adjuvant chemotherapy was given in all 53(100%) cases, whereas neo-adjuvant radiation therapy was offered to 49(92.5%) patients. There was a significant and favorable patient response to the neo-adjuvant treatment in 37(69.8%) cases, leading to a decrease in tumor size. Laparoscopic McKeown Esophagectomies were performed in 44 (83.0%) and 9(17.0%) were Robot-assisted Minimally Invasive esophagectomy (RAMIE). Intraoperative injuries (i.e., lung parenchymal injury and bleeding) were reported in only 2(3.8%) patients. Post-operative complications were recorded in 12(22.6%) patients. Margin clearance was observed in 53 (100%) of the patients. The 90-day mortality rate was 3(5.7%), one due to bleeding and other two mortalities were due to COVID related respiratory complications. Minimally invasive esophagectomy was found to be safe and feasible technique with encouraging results in terms of decreased intraoperative and post operative complications as well as achieving the standard oncological surgery with acceptable lymph node yield and margin clearance and in hospital and 90 days mortality.


Dynamic Changes in Heparan Sulfate Nanostructure in Human Pluripotent Stem Cell Differentiation.
Heparan sulfate (HS) is a heterogeneous, cell-surface polysaccharide critical for transducing signals essential for mammalian development. Imaging of signaling proteins has revealed how their localization influences their information transfer. In contrast, the contribution of the spatial distribution and nanostructure of information-rich, signaling polysaccharides like HS is not known. Using expansion microscopy (ExM), we found striking changes in HS nanostructure occur as human pluripotent stem (hPS) cells differentiate, and these changes correlate with growth factor signaling. Our imaging studies show that undifferentiated hPS cells are densely coated with HS displayed as hair-like protrusions. This ultrastructure can recruit fibroblast growth factor for signaling. When the hPS cells differentiate into the ectoderm lineage, HS is localized into dispersed puncta. This striking change in HS distribution coincides with a decrease in fibroblast growth factor binding to neural cells. While developmental variations in HS sequence were thought to be the primary driver of alterations in HS-mediated growth factor signaling, our high-resolution images indicate a role for the HS nanostructure. Our study highlights the utility of high-resolution glycan imaging using ExM. In the case of HS, we found that changes in how the polysaccharide is displayed link to profound differences in growth factor binding.


The effects of induced emotions on environmental preferences and behavior: An experimental study.
Communication policies employed by policymakers and non-governmental organizations (NGOs) often appeal to the emotions to persuade people to adopt virtuous behavior. The aim of this paper is to study the impact of induced emotions on pro-environmental behavior (PEB). We design a three-stage laboratory experiment. In the first stage, we determine the level of the subjects' environmental awareness. In the second stage, subjects read scripts that place them in realistic hypothetical scenarios designed to induce specific emotions. We implement a 2 x 2 in-between design by varying both the valence and social dimension of the four emotional states induced: happiness, sadness, pride and shame. In the third stage, subjects play a modified dictator game in which the recipient is an environmental non-governmental organization (ENGO). We show that the emotional states of subjects can influence PEB. In particular, negative emotions significantly reduce the average individual amount of donations made to ENGOs. We also find that the precise impact of the emotional states is more complex and appears to be dependent on individuals' characteristics and awareness for environmental issues. For instance, in positive emotional states, men donate significantly less than women. In addition, a high level of environmental awareness increases donations in subjects experiencing shame and decreases their likelihood to donate when feeling pride. Also, we observe behavioral consistency for negative emotions and rather compensatory behavior for positive emotions.


The Efficacy of Silver-Based Electrospun Antimicrobial Dressing in Accelerating the Regeneration of Partial Thickness Burn Wounds Using a Porcine Model.
(1) Background: Wounds with damages to the subcutaneous are difficult to regenerate because of the tissue damages and complications such as bacterial infection. (2) Methods: In this study, we created burn wounds on pigs and investigated the efficacy of three biomaterials: polycaprolactone-gelatin-silver membrane (PCLGelAg) and two commercial burn dressings, Aquacel® Ag and UrgoTulTM silver sulfadiazine. In vitro long-term antibacterial property and in vivo wound healing performance were investigated. Agar diffusion assays were employed to evaluate bacterial inhibition at different time intervals. Minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and time-kill assays were used to compare antibacterial strength among samples. Second-degree burn wounds in the pig model were designed to evaluate the efficiency of all dressings in supporting the wound healing process. (3) Results: The results showed that PCLGelAg membrane was the most effective in killing both Gram-positive and Gram-negative bacteria bacteria with the lowest MBC value. All three dressings (PCLGelAg, Aquacel, and UrgoTul) exhibited bactericidal effect during the first 24 h, supported wound healing as well as prevented infection and inflammation. (4) Conclusions: The results suggest that the PCLGelAg membrane is a practical solution for the treatment of severe burn injury and other infection-related skin complications.


Analysis of Interaction Between Interfacial Structure and Fibrinogen at Blood-Compatible Polymer/Water Interface.
The correlation between the interfacial structure and protein adsorption at a polymer/water interface was investigated. Poly(2-methoxyethyl acrylate)(PMEA), which is one of the best blood compatible polymers available, was employed. Nanometer-scale structures generated through the phase separation of polymer and water were observed at the PMEA/phosphate buffered saline interface. The interaction between the interfacial structures and fibrinogen (FNG) was measured using atomic force microscopy. Attraction was observed in the polymer-rich domains as well as in the non-blood compatible polymer. In contrast, no attractive interactions were observed, and only a repulsion occurred in the water-rich domains. The non-adsorption of FNG into the water rich domains was also clarified through topographic and phase image analyses. Furthermore, the FNG molecules adsorbed on the surface of PMEA were easily desorbed, even in the polymer-rich domains. Water molecules in the water-rich domains are anticipated to be the dominant factor in preventing FNG adsorption and thrombogenesis on a PMEA interface.


[Effect of a new anti-inflammatory agent chlotazol on nucleic acid metabolism in rat liver cells].
Effects of a new antiinflammatory preparation chlotozol on the activity of enzymes participating in metabolism of nucleic acids, on rates of DNA and RNA synthesis and on the transport of RNA from nuclei into cytoplasm were studied in liver cells of Wistar rats. Administration of chlotozol did not distinctly affect the rate of DNA synthesis, the RNAase activity, the rate of RNA transport from nuclei into cytoplasm. Chlotozol caused a decrease of the RNA polymerase activity in nuclei in vitro. Thus, chlotozol did not exhibit negative effects on reduplication and transport of RNA from nuclei into cytoplasm in liver cells of intact animals. Inhibitory effect of chlotozol on the RNA-polymerase activity appears to be one of mechanisms responsible for regulation of transcription.


The Preferred Retinal Locus Used to Watch Videos.
Eccentric viewing is a common strategy used by people with central vision loss (CVL) to direct the eye such that the image falls onto functioning peripheral retina, known as the preferred retinal locus (PRL). It has been long acknowledged that we do not know whether the PRL used in a fixation test is also used when performing tasks. We present an innovative method to determine whether the same PRL observed during a fixation task was used to watch videos and whether poor resolution affects gaze location. The gaze of a group of 60 normal vision (NV) observers was used to define a democratic center of interest (COI) of video clips from movies and television. For each CVL participant (N = 20), we computed the gaze offsets from the COI across the video clips. The distribution of gaze offsets of the NV participants was used to define the limits of NV behavior. If the gaze offset was within this 95% degree confidence interval, we presumed that the same PRL was used for fixation and video watching. Another 15 NV participants watched the video clips with various levels of defocus blur. CVL participants had wider gaze-offset distributions than NV participants (P < 0.001). Gaze offsets of 18/20 CVL participants were outside the NV confidence interval. Further, none of the 15 NV participants watching the same videos with spherical defocus blur had a gaze offset that was decentered (outside the NV confidence interval), suggesting that resolution was not the problem. This indicates that many CVL participants were using a PRL to view videos that differed from that found with a fixation task and that it was not caused by poor resolution alone. The relationship between these locations needs further investigation.


Gibberellin concentration and transport in genetic lines of pea : effects of grafting.
Effects of the Na and Le loci on gibberellin (GA) content and transport in pea (Pisum sativum L.) shoots were studied. GA(1), GA(8), GA(17), GA(19), GA(20), GA(29), GA(44), GA(8) catabolite, and GA(29) catabolite were identified by full-scan gas chromatography-mass spectrometry in extracts of expanding and fully expanded tissues of line C79-338 (Na Le). Quantification of GAs by gas chromatography-single-ion monitoring using deuterated internal standards in lines differing at the Na and Le alleles showed that na reduced the contents of GA(19), GA(20), and GA(29) on average to <3% and of GA(1) and GA(8) to <30% of those in corresponding Na lines. In expanding tissues from Na le lines, GA(1) and GA(8) concentrations were reduced to approximately 10 and 2%, respectively, and GA(29) content increased 2- to 3-fold compared with those in Na Le plants. There was a close correlation between stem length and the concentrations of GA(1) or GA(8) in shoot apices in all six genotypes investigated. In na/Na grafts, internode length and GA(1) concentration of nana scions were normalized, the GA(20) content increased slightly, but GA(19) levels were unaffected. Movement of labeled GAs applied to leaves on Na rootstocks indicated that GA(19) was transported poorly to apices of na scions compared with GA(20) and GA(1). Our evidence suggests that GA(20) is the major transported GA in peas.


Non-Traumatic Subdural Hemorrhage and Risk of Arterial Ischemic Events.
Background and Purpose- The risk of arterial ischemic events after subdural hemorrhage (SDH) is poorly understood. This study aimed to evaluate the risk of acute ischemic stroke and myocardial infarction among patients with and without nontraumatic SDH. Methods- We performed a retrospective cohort study using claims data from 2008 through 2014 from a nationally representative sample of Medicare beneficiaries. The exposure was nontraumatic SDH. Our primary outcome was an arterial ischemic event, a composite of acute ischemic stroke and acute myocardial infarction. Secondary outcomes were ischemic stroke alone and myocardial infarction alone. We used validated International Classification of Diseases, Ninth Revision, Clinical Modification diagnosis codes to identify our predictor and outcomes. Using Cox regression and corresponding survival probabilities, adjusted for demographics and vascular comorbidities, we computed the hazard ratio in 4-week intervals after SDH discharge. We performed secondary analyses stratified by strong indications for antithrombotic therapy (composite of atrial fibrillation, peripheral vascular disease, valvular heart disease, and venous thromboembolism). Results- Among 1.7 million Medicare beneficiaries, 2939 were diagnosed with SDH. In the 4 weeks after SDH, patients' risk of an arterial ischemic event was substantially increased (hazard ratio, 3.6 [95% CI, 1.9-5.5]). There was no association between SDH diagnosis and arterial ischemic events beyond 4 weeks. In secondary analysis, during the 4 weeks after SDH, patients' risk of ischemic stroke was increased (hazard ratio, 4.2 [95% CI, 2.1-7.3]) but their risk of myocardial infarction was not (hazard ratio, 0.8 [95% CI, 0.2-1.7]). Patients with strong indications for antithrombotic therapy had increased risks for arterial ischemic events similar to patients in the primary analysis, but those without such indications did not demonstrate an increased risk for arterial ischemic events. Conclusions- Among Medicare beneficiaries, we found a heightened risk of arterial ischemic events driven by an increased risk of ischemic stroke, in the 4 weeks after nontraumatic SDH. This increased risk may be due to interruption of antithrombotic therapy after SDH diagnosis.


Thromboembolic disease in vasculitis.
To give an overview of recent clinical findings of thromboembolic disease in vasculitis and provide insight into possible explanations of the association between thrombosis and inflammation. A high incidence of venous thrombotic events has recently been described in four distinct cohorts of patients with antineutrophil cytoplasmic antibody-associated vasculitis (AAV), especially during periods of active disease. No factors other than the vasculitis itself have been identified that explain this high occurrence of thrombosis. Several studies have shown an increased rate of thrombosis in Behçet's disease, with a different clinical presentation from that observed in AAV. Recent laboratory findings provide exciting insights into a bidirectional feedback loop between coagulation and inflammation that may be applicable to vasculitis. Thrombosis is an important clinical manifestation of some types of vasculitis. Better understanding of the association of thrombosis with inflammation in vasculitis might lead to development of clinically useful biomarkers and new approaches to therapy. Additionally, study of the specific factors involved in thrombosis in systemic vasculitis could help explain the role of inflammation in more common settings of venous thrombotic events.


The active zone protein family ELKS supports Ca2+ influx at nerve terminals of inhibitory hippocampal neurons.
In a presynaptic nerve terminal, synaptic vesicle exocytosis is restricted to specialized sites called active zones. At these sites, neurotransmitter release is determined by the number of releasable vesicles and their probability of release. Proteins at the active zone set these parameters by controlling the presynaptic Ca(2+) signal, and through docking and priming of synaptic vesicles. Vertebrate ELKS proteins are enriched at presynaptic active zones, but their functions are not well understood. ELKS proteins are produced by two genes in vertebrates, and each gene contributes ∼50% to total brain ELKS. We generated knock-out mice for ELKS1 and found that its constitutive removal causes lethality. To bypass lethality, and to circumvent redundancy between ELKS1 and ELKS2 in synaptic transmission, we used a conditional genetic approach to remove both genes in cultured hippocampal neurons after synapses are established. Simultaneous removal of ELKS1 and ELKS2 resulted in a 50% decrease of neurotransmitter release at inhibitory synapses, paralleled by a reduction in release probability. Removal of ELKS did not affect synapse numbers or their electron microscopic appearance. Using Ca(2+) imaging, we found that loss of ELKS caused a 30% reduction in single action potential-triggered Ca(2+) influx in inhibitory nerve terminals, consistent with the deficits in synaptic transmission and release probability. Unlike deletion of the active zone proteins RIM, RIM-BP, or bruchpilot, ELKS removal did not lead to a measurable reduction in presynaptic Ca(2+) channel levels. Our results reveal that ELKS is required for normal Ca(2+) influx at nerve terminals of inhibitory hippocampal neurons.


Tuberculosis of the foot and ankle.
Tuberculosis of the foot and ankle is an uncommon presentation of skeletal tuberculosis. The uncommon site, lack of awareness, and ability to mimic other disorders clinically and on radiographs, leads to diagnostic and therapeutic delays. In the early stages and when the disease is limited to bone, medical treatment leads to excellent healing and limited residual disabilities. Joint involvement occurs because of spread from a periarticular bony focus, and in the midfoot, the disease may spread to involve all the interconnected joints, leading to a stiff foot and residual deformities. Of 74 cases reviewed, the calcaneus was involved most commonly, followed by infection of the midtarsal, Lisfranc joints, and ankle. The most common radiologic finding is that of osteoporosis, which may be intense: cancellous bone involvement may present as a cystic lesion with or without sequestrum. Computed tomography scans and magnetic resonance imaging are helpful examinations. Because the disease is paucibacillary, a positive acid fast bacilli culture is rare and the diagnosis usually is confirmed by obtaining granulomatous tissue on biopsy. The treatment basically is medical, with surgical intervention being reserved for patients with intractable disease or as a salvage procedure for patients with deformed hindfoot joints.


From 1,2-difunctionalisation to cyanide-transfer cascades - Pd-catalysed cyanosulfenylation of internal (oligo)alkynes.
Internal alkynes substituted by aliphatic or aromatic moieties or by heteroatoms were converted into sulphur-substituted acrylonitrile derivatives. Key is the use of Pd catalysis, which allows the addition of aromatic and aliphatic thiocyanates in an intra- and intermolecular manner. Substrates with several alkyne units underwent further carbopalladation steps after the initial thiopalladation step, thus generating in a cascade-like fashion an oligoene unit with sulphur at one terminus and the cyano group at the other.


Differential effect of adriamycin on DNA replicative and repair synthesis in cultured neonatal rat cardiac cells.
The effect of the potent antitumor antiobiotic Adriamycin (ADM) on DNA replication and unscheduled DNA synthesis in cultured rat cardiac cells was investigated. Autoradiography and [3H]thymidine incorporation studies were carried out on parallel cultures. DNA replication was depressed for up to 6 days following a 3-hr pulse of ADM administration. An ADM concentration of 1 microgram/ml which was effective in reducing replicative DNA synthesis by as much as 75% did not reduce the ability of cardiac cells to repair UV-damaged DNA. However, cells exposed to higher ADM concentrations failed to undergo significant UV-induced repair. In the absence of UV treatment, ADM did not stimulate unscheduled DNA synthesis. To account for the differential response of the cardiac cell cultures to replicate and repair DNA, we propose that ADM exerts a localized effect on DNA synthesis covering a region proximal to its primary intercalation site.


Carnitine palmitoyltransferase in liver and five extrahepatic tissues in the rat. Inhibition by DL-2-bromopalmitoyl-CoA and effect of hypothyroidism.
Mitochondria were isolated from rat adult liver, foetal liver, kidney cortex, heart, skeletal muscle and interscapular brown adipose tissue. DL-2-Bromopalmitoyl-CoA inhibited the overt form of carnitine palmitoyltransferase (CPT1) in heart, skeletal muscle and brown adipose tissue, with an IC50 value (concentration giving 50% inhibition) of 1.3-1.6 microM. By contrast, the IC50 value for inhibition of the kidney or adult liver enzyme was 0.08-0.1 microM. CPT1 in near-term foetal liver differed from that in adult liver in that the IC50 for inhibition by 2-bromopalmitoyl-CoA was 0.57 microM. It is suggested that there may be tissue-specific forms of the catalytic entity of CPT1 and that foetal liver may contain a mixture of adult liver- and muscle-type enzymes. In rats made hypothyroid by administration of propylthiouracil and an iodine-deficient diet, hepatic CPT1 activity was decreased by 83%. However, CPT1 activity in extrahepatic tissues showed no adaptive decrease in hypothyroidism.


Thermodynamic basis for the demarcation of Arctic and alpine treelines.
At the edge of alpine and Arctic ecosystems all over the world, a transition zone exists beyond which it is either infeasible or unfavorable for trees to exist, colloquially identified as the treeline. We explore the possibility of a thermodynamic basis behind this demarcation in vegetation by considering ecosystems as open systems driven by thermodynamic advantage-defined by vegetation's ability to dissipate heat from the earth's surface to the air above the canopy. To deduce whether forests would be more thermodynamically advantageous than existing ecosystems beyond treelines, we construct and examine counterfactual scenarios in which trees exist beyond a treeline instead of the existing alpine meadow or Arctic tundra. Meteorological data from the Italian Alps, United States Rocky Mountains, and Western Canadian Taiga-Tundra are used as forcing for model computation of ecosystem work and temperature gradients at sites on both sides of each treeline with and without trees. Model results indicate that the alpine sites do not support trees beyond the treeline, as their presence would result in excessive CO[Formula: see text] loss and extended periods of snowpack due to temperature inversions (i.e., positive temperature gradient from the earth surface to the atmosphere). Further, both Arctic and alpine sites exhibit negative work resulting in positive feedback between vegetation heat dissipation and temperature gradient, thereby extending the duration of temperature inversions. These conditions demonstrate thermodynamic infeasibility associated with the counterfactual scenario of trees existing beyond a treeline. Thus, we conclude that, in addition to resource constraints, a treeline is an outcome of an ecosystem's ability to self-organize towards the most advantageous vegetation structure facilitated by thermodynamic feasibility.


Antigen induced arthritis in beige (Chediak-Higashi) mice.
Mice with the beige mutation, which are known to be deficient for leucocyte elastase and cathepsin G, were used to investigate the role of neutral proteases in a model for antigen induced arthritis. Surprisingly, it was shown that in this model of arthritis, using methylated bovine serum albumin as an antigen, C57/black/6 'beige' mice (deficient for leucocyte neutral proteases) developed a more severe form of arthritis than the control mice ('black' mice), resulting in a higher degree of tissue damage. The incidence and degree of bone apposition and destruction of articular cartilage at day 21 after induction of arthritis were significantly higher in the beige mice. These findings could not be ascribed to differences in the cellular immune response to methylated bovine serum albumin. Autoradiographic detection of radiolabelled methylated bovine serum albumin suggested that more antigen is retained in the joints of beige mice than in black mice, which might account for the sustained arthritis and the concomitant tissue damage. These findings do not support the contention that leucocyte elastase and cathepsin G contribute to the pathogenesis of joint destruction in this model.


[Current status of the SAPHO syndrome].
In 1987, our group proposed the acronym SAPHO (Synovitis Acne Pustulosis Hyperostosis Osteitis) to identify a clinical entity with characteristic manifestations formerly described under a wide variety of names. Since our first description, several new points have been recognized. For example it has been demonstrated that the skin lesions are not necessarily concomitant with osteoarticular manifestations sometimes separated by a long time interval. There are also several arguments suggesting a relationship with spondyloarthropathy. In addition, pseudo-tumoral fibrosis of anterior thoracic lesions which may lead to venous compression have been observed. It is difficult to determine the frequency of SAPHO. Most of the cases reported were observed in western Europe or Japan. Now that the concept has been accepted by our British colleagues, the number of cases reported has increased steadily, perhaps suggesting a reason for the apparently low frequency in the United States. The pathogenesis is still unknown. There is some evidence however that Propionibacterium acnes infection may be involved, at least in some cases, but formal proof is still lacking. Treating patients with SAPHO thus remains a question of debate. Non-steroid antiinflammatory agents are currently the treatment of choice although certain authors have tried colchicine and sulfasalazine or methotrexate with little success. There remains a good deal of work to be done before we thoroughly understand the SAPHO syndrome but we can nevertheless assure our patients that this strange bone disease is neither a tumour nor an infection and is never severely invalidating.


Portable Anterior Eye Segment Imaging System for Teleophthalmology.
This study aims to compare a new prototype for a portable anterior eye segment imaging system with the standard method for ophthalmology examination. The new imaging system consisted of two IMX219 Arducam autofocus sensors (Arducam, China, Nanjing) for Raspberry Pi V2 camera module connected to a Raspberry Pi Zero W (Raspberry Pi Foundation, UK, Cambridge) that clips to a wearable headset. The 2D videos of the anterior eye segment were recorded with the new system and a 720p FaceTime HD camera (Apple, Cupertino, CA). Afterward, ophthalmologists evaluated the videos using a standard clinical eye examination form. These evaluations were compared with the standard slit-lamp clinical assessment performed during the patient's visit. Thirty-five eyes were evaluated. The sensitivity and specificity percentages were statistically significant between the two imaging modalities (P ≤ 0.001). The evaluations performed from videos obtained with the new imaging system had better sensitivity and specificity percentages overall. However, statistically significant differences were only observed in cornea, anterior chamber, iris, and lens. Specificity percentages were higher than sensitivity percentages in both imaging modalities, indicating that video evaluations are less accurate for pathological screening. Nevertheless, the new system evaluations were significantly better than the webcam evaluations. This study presented an alternative system to assess eye conditions for telemedicine, one that provides more details than the current standard and uses new wearable headsets technologies.


[New imaging methods for visualizing coronary arteries].
Techniques in the field of coronary artery imaging can be divided into two groups: invasive and non-invasive methods. Apart from the conventional coronary artery angiography, invasive methods include intracoronary ultrasound, intracoronary angioscopy, and optical coherence tomography. Non-invasive methods include magnetic resonance tomography, synchrotron-coronary angiography, and electron beam computed tomography. In the late 1980s, intracoronary ultrasound has come into clinical practice. It offers a real-time, cross-sectional image of the coronary artery in high resolution. Coronary arteries enlarge in the presence of atherosclerotic plaque formation in order to compensate for luminal narrowing caused by plaque formation (remodeling). With coronary angiography, the plaque formation cannot be detected until a lumen reduction of about 40-45%. With intravascular ultrasound, the early stages of atherosclerosis can clearly be demonstrated. In combination with the intracoronary Doppler technique, syndrome X can be differentiated. Another important role of intracoronary ultrasound in the diagnosis of coronary artery disease is to guide coronary interventions and to assess the result of coronary interventions especially to evaluate the result of stent implantation. Due to the clinical use of intracoronary ultrasound and the guidance of high pressure stent implantation, the incidence of acute stent thrombosis has decreased to about 1%. Coronary angioscopy portrays the surface of the vessel lumen. It is helpful to identify the mural thrombus especially to differentiate fresh and chronic thrombus formation. Magnetic resonance tomography is able to image the coronary arterial contour of the proximal segment. With today's gating technique, it is possible to portray the whole coronary tree and avoid disturbances resulting from the heart beat and respiration. Electron beam computed tomography is a very promising technique in screening for coronary artery disease. It is a very sensitive method to identify coronary calcification and, thus, to detect atherosclerotic plaque. Studies have shown that the presence of calcification almost invariably indicates the presence of coronary artery disease and that the absence of calcification can nearly rule out significant coronary artery disease. Moreover, a close correlation exists between the amount of calcification and the severity of coronary artery disease. Additionally, in combination with contrast injection, coronary artery perfusion can be evaluated. This is important to assess the conductance of coronary stent and bypass graft.


Tamoxifen-mediated growth inhibition of human cholangiocarcinoma.
Cholangiocarcinoma represents a challenging primary malignancy of the liver with no effective medical therapy and a poor prognosis. We have investigated the role of tamoxifen and estrogen receptors (ERs) in the regulation of growth of human cholangiocarcinoma. Two human cholangiocarcinoma cell lines, OZ and SK-ChA-1, were grown in the presence of graded concentrations of tamoxifen; the effects on cell growth were determined by cell counting or 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium proliferation assay. The presence of ER protein was tested by indirect immunofluorescence and immunoprecipitation. In addition, cells were grown in estrogen-depleted media supplemented with exogenous 17beta-estradiol. ER mRNA was evaluated by reverse transcription-PCR and Northern blotting. Finally, one cholangiocarcinoma cell line was grown as a xenograft in athymic nude mice; tamoxifen effects on in vivo tumor growth were determined with biweekly caliper measurements. Tamoxifen (5-10 microM) caused dose-dependent in vitro growth inhibition of two human cholangiocarcinoma cell lines. In addition, growth inhibition of one cell line (SK-ChA-1) grown as a xenograft in nude mice by tamoxifen was observed. The presence of ER protein was suggested by 17beta-estradiol stimulation of tumor cell growth in vitro and confirmed by immunoprecipitation. Immunofluorescence microscopy was ineffective at detection of ER protein. Reverse transcription-PCR demonstrated the presence of ER mRNA in both cell lines. Northern blot analysis confirmed the presence of full-length 6.5-kb ER mRNA. No ER deletion mutants were detected. Tamoxifen inhibited the growth of human cholangiocarcinoma in vitro and in vivo. ER protein and mRNA were detected in both cell lines. The mechanism(s) of tamoxifen-mediated growth inhibition is unclear but may occur via ER protein or additional pathways. The ability of tamoxifen to inhibit tumor growth may offer an alternative adjunctive treatment for cholangiocarcinoma.


Gelatinase B-deficient mice are resistant to experimental bullous pemphigoid.
Bullous pemphigoid (BP) is an autoimmune subepidermal blistering disease characterized by deposition of autoantibodies at the basement membrane zone. In an experimental BP model in mice, the subepidermal blistering is mediated by antibodies directed against the hemidesmosomal protein BP180 (collagen XVII, BPAG2), and depends on complement activation and neutrophil infiltration. Gelatinase B is present in BP blister fluid and can cleave BP180. In this study we investigated the role of gelatinase B in the immunopathogenesis of experimental BP using mice containing targeted disruption of the gelatinase B (MMP-9, 92 kD gelatinase) gene. Gelatinase B-deficient mice were resistant to the blistering effect of intracutaneous anti-mBP180 antibodies, although these mice showed deposition of autoantibodies at the basement membrane zone and neutrophil recruitment to the skin comparable to that observed in the control mice. Interleukin 8 given intradermally concomitantly with pathogenic anti-mBP180 elicited a significant neutrophil recruitment into the skin in gelatinase B-deficient mice, but blistering did not occur. However, gelatinase B-deficient mice reconstituted with neutrophils from normal mice developed blistering in response to anti-mBP180 antibodies. These results implicate neutrophil-derived gelatinase B in the pathogenesis of experimental BP and might lead to novel therapeutic strategies for BP.


Cigarette use among Canadian undergraduates.
To describe the prevalence of daily cigarette use among Canadian undergraduates. Estimates are also compared to earlier Ontario surveys. Data are drawn from the Canadian Campus Survey, a national mail survey, conducted in the fall of 1998, with a random sample of 7,800 students from 16 universities. Overall, 17.1% reported daily cigarette smoking and 10.4% reported occasional smoking. Rates of daily smoking differed significantly by region (with rates above average among those attending university in the Atlantic and below average among those attending university in British Columbia and the Prairies), residence (those residing off campus without family reported the highest prevalence rate), and year of study (those in the final year typically reported lower rate of use). University campuses represent an environment with potential gains to be made by tobacco control policies.


Idiopathic syrinx in the pediatric population: a combined center experience.
Discovery of a syrinx in a child, without a readily identifiable proximate cause such as a Chiari malformation, tumor, or site of tethering, is often a cause of concern for families and a source of consternation for clinicians. There is a paucity of data describing the natural history of an idiopathic syrinx in the pediatric population. The authors present the combined data of 2 major pediatric neurosurgical centers to describe their experience with this condition. Data were collected at Children's Hospital Boston and St. Louis Children's Hospital according to institutional review board-approved protocols and captured visits over a 2.5-year interval (October 2006-March 2009), with records reviewed if the patient had a preexisting diagnosis of syrinx. Patients were identified by ICD-9 codes derived from departmental databases. All pediatric patients (age < 19 years) in whom idiopathic syrinx had been diagnosed, as defined by MR imaging findings (dilated central canal in the spinal cord of ≥ 1 mm in axial dimension and extending over at least 2 vertebral levels), were included. Forty-eight patients met the criteria for idiopathic syrinx during this period, and in 32 of them detailed follow-up imaging was available. Discovery of a syrinx was incidental in 6 patients, whereas the others were referred for imaging because of the presence of pain, neurological symptoms, scoliosis, or skin markings. The average age at the first MR imaging session was 9.7 years, with a mean syrinx size of 4 mm (range 1.2-9.4 mm). The majority (52%) of patients had a thoracic syrinx, with the average lesion spanning 7.1 vertebral levels. The average follow-up was 23.8 months (range 2-64 months), and subgroups of patients with < 3 years and ≥ 3 years of follow-up were independently reviewed. Overall, symptoms improved in 34% and worsened in 9%; 57% of the patients remained asymptomatic or stable. Radiographically (in the subgroup of 32 patients with detailed follow-up imaging), syrinx size decreased in 25% of patients, increased in 12.5%, and remained unchanged in 62.5%, with no apparent correlation between change in syrinx size and clinical symptoms. Clinically, children with an idiopathic syrinx remained asymptomatic, stable, or improved in 91% of cases. The majority of syringes (87.5%) remained stable or shrank over time, with no apparent correlation between changes in size and changes in symptoms. Although longer follow-up is needed, these data suggest that the natural history of an idiopathic syrinx in children is benign, and that repeated imaging may not be necessary.


Effect of Yoga-Based Cardiac Rehabilitation on Heart Rate Variability: Randomized Controlled Trial in Patients Post-MI.
Autonomic dysfunction is an independent predictor of cardiovascular and all-cause mortality after myocardial infarction (MI). We tested the effects of a 12-week yoga-based cardiac rehabilitation program on heart rate variability (HRV) in 80 patients post-MI. This randomized controlled trial with two parallel groups was carried out in a tertiary care institution in India. The yoga group received 13 hospital-based structured yoga sessions as an adjunct to standard care. Control group participants received enhanced standard care involving three brief educational sessions with a leaflet on the importance of diet and physical activity. HRV was measured in all participants with lead II electrocardiogram (ECG) signals. One yoga group patient's data were excluded due to ECG abnormalities. Baseline measurement was done 3 weeks post-MI, and postintervention assessment took place at the 13th week. HRV frequency and time domain indices were analyzed. There were no significant between-group differences in the HRV time domain indices. Frequency domain indices showed significant between-group differences in HF power (absolute) (yoga vs. control: 114.42 [-794.80-7,993.78] vs. -38.14 [-4,843.50-1,617.87], p = 0.005) and total power (nu) (yoga vs. control: 44.96 [21.94] vs. -19.55 [15.42], p = 0.01) with higher HF power and total power (nu) in the yoga group. It should be noted that these results cannot be generalized to high risk patients. Respiratory frequency control to check for influence of respiratory rate on RR interval was not evaluated. This short-term yoga-based cardiac rehabilitation program had additive effects in shifting sympathovagal balance toward parasympathetic predominance while increasing overall HRV in optimally medicated post-MI patients.


Temporal trends in Enterobacter species bloodstream infection: a population-based study from 1998-2007.
Enterobacter species are the fourth most common cause of Gram-negative bloodstream infection (BSI). We examined temporal changes and seasonal variation in the incidence rate of Enterobacter spp. BSI, estimated 28-day and 1-year mortality, and determined in vitro antimicrobial resistance rates of Enterobacter spp. bloodstream isolates in Olmsted County, Minnesota, from 1 January 1998 to 31 December 2007. Multivariable Poisson regression was used to examine temporal changes and seasonal variation in incidence rate and Kaplan-Meier method was used to estimate 28-day and 1-year mortality. The median age of patients with Enterobacter spp. BSI was 58 years and 53% were female. The overall age- and gender-adjusted incidence rate of Enterobacter spp. BSI was 3.3 per 100,000 person-years (95% CI 2.3-4.4). There was a linear trend of increasing incidence rate from 0.8 (95% CI 0-1.9) to 6.2 (95% CI 3.0-9.3) per 100,000 person-years between 1998 and 2007 (p 0.002). There was no significant difference in the incidence rate of Enterobacter spp. BSI during the warmest 4 months compared to the remainder of the year (incidence rate ratio 1.06; 95% CI 0.47-2.01). The overall 28-day and 1-year mortality rates of Enterobacter spp. BSI were 21% (95% CI 8-34%) and 38% (95% CI 22-53%), respectively. Up to 13% of Enterobacter spp. bloodstream isolates were resistant to third-generation cephalosporins. To our knowledge, this is the first population-based study to describe the epidemiology and outcome of Enterobacter spp. BSI. The increase in incidence rate of Enterobacter spp. BSI over the past decade, coupled with its associated antimicrobial resistance, dictate the need for further investigation of this syndrome.


Validation of the Wave 1 and Wave 2 Population Assessment of Tobacco and Health (PATH) Study Indicators of Tobacco Dependence Using Biomarkers of Nicotine Exposure Across Tobacco Products.
This study examined the predictive relationships between biomarkers of nicotine exposure and 16-item self-reported level of tobacco dependence (TD) and subsequent tobacco use outcomes. The Population Assessment of Tobacco and Health (PATH) Study surveyed adult current established tobacco users who provided urine biospecimens at Wave 1 (September 2013-December 2014) and completed the Wave 2 (October 2014-October 2015) interview (n = 6872). Mutually exclusive user groups at Wave 1 included: Cigarette Only, E-cigarette Only, Cigar Only, Hookah Only, Smokeless Tobacco Only, Cigarette Plus E-cigarette, multiple tobacco product users who smoked cigarettes, and multiple tobacco product users who did not smoke cigarettes. Total Nicotine Equivalents (TNE-2) and TD were measured at Wave 1. Approximate one-year outcomes included frequency/quantity used, quitting, and adding/switching to different tobacco products. For Cigarette Only smokers and multiple tobacco product users who smoked cigarettes, higher TD and TNE-2 were associated with: a tendency to smoke more, smoking more frequently over time, decreased likelihood of switching away from cigarettes, and decreased probability of quitting after one year. For other product user groups, Wave 1 TD and/or TNE-2 were less consistently related to changes in quantity and frequency of product use, or for adding or switching products, but higher TNE-2 was more consistently predictive of decreased probability of quitting. Self-reported TD and nicotine exposure assess common and independent aspects of dependence in relation to tobacco use behaviors for cigarette smokers. For other product user groups, nicotine exposure is a more consistent predictor of quitting than self-reported TD. This study suggests that smoking cigarettes leads to the most coherent pattern of associations consistent with a syndrome of TD. Because cigarettes continue to be prevalent and harmful, efforts to decrease their use may be accelerated via conventional means (eg, smoking cessation interventions and treatments), but also perhaps by decreasing their dependence potential. The implications for noncombustible tobacco products are less clear as the stability of tobacco use patterns that include products such as e-cigarettes continue to evolve. TD, nicotine exposure measures, and consumption could be used in studies that attempt to understand and predict product-specific tobacco use behavioral outcomes.


Assessing the Needs and Perspectives of Patients With Asthma and Chronic Obstructive Pulmonary Disease on Patient Web Portals: Focus Group Study.
As accessibility to the internet has increased in society, many health care organizations have developed patient Web portals (PWPs), which can provide a range of self-management options to improve patient access. However, the available evidence suggests that they are used inefficiently and do not benefit patients with low health literacy. Asthma and chronic obstructive pulmonary disease (COPD) are common chronic diseases that require ongoing self-management. Moreover, patients with COPD are typically older and have lower health literacy. This study aimed to obtain and present an overview of patients' perspectives of PWPs to facilitate the development of a portal that better meets the needs of patients with asthma and COPD. We performed a focus group study using semistructured interviews in 3 patient groups from the north of the Netherlands who were recruited through the Dutch Lung Foundation. Each group met 3 times for 2 hours each at a 1-week interval. Data were analyzed with coding software, and patient descriptors were analyzed with nonparametric tests. The consolidated criteria for reporting qualitative research were followed when conducting the study. We included 29 patients (16/29, 55% male; mean age 65 [SD 10] years) with COPD (n=14), asthma-COPD overlap (n=4), asthma (n=10), or other respiratory disease (n=1). There was a large variation in the internet experience; some patients hardly used the internet (4/29, 14%), whereas others used internet >3 times a week (23/29, 79%). In general, patients were positive about having access to a PWP, considering access to personal medical records as the most important option, though only after discussion with their physician. A medication overview was considered a useful option. We found that communication between health care professionals could be improved if patients could use the PWP to share information with their health care professionals. However, as participants were worried about the language and usability of portals, it was recommended that language should be adapted to the patient level. Another concern was that disease monitoring through Web-based questionnaire use would only be useful if the results were discussed with health care professionals. Participants were positive about PWPs and considered them a logical step. Today, most patients tend to be better educated and have internet access, while also being more assertive and better informed about their disease. A PWP could support these patients. Our participants also provided practical suggestions for implementation in current and future PWP developments. The next step will be to develop a portal based on these recommendations and assess whether it meets the needs of patients and health care providers.


[Biosynthesis of L-asparaginase-2 by cultures of Bacillus polymyxa var. Ross].
Cell extracts of Bacillus polymyxa var. Ross.--producer of the polypeptide antibiotic polymyxin M. showed activity of L-asparaginase-2 (L-asparagine aminohydrolase EC 126.96.36.199). The enzyme activity in the growing culture increased with the biomass. The highest specific activity was detected in the cells at the onset of the stationary stage. The synthesis of L-asparaginase-2 was subjected to glucose catabolite repression in response to its addition to the culture at the logarithmic stage. After purification L-asparaginase-2 was obtained that was 350 times more active than the initial preparation. The enzyme properties were examined.


QEEG and LORETA in Teenagers With Conduct Disorder and Psychopathic Traits.
Few studies have investigated the impact of the psychopathic traits on the EEG of teenagers with conduct disorder (CD). To date, there is no other research studying low-resolution brain electromagnetic tomography (LORETA) technique using quantitative EEG (QEEG) analysis in adolescents with CD and psychopathic traits. To find electrophysiological differences specifically related to the psychopathic traits. The current investigation compares the QEEG and the current source density measures between adolescents with CD and psychopathic traits and adolescents with CD without psychopathic traits. The resting EEG activity and LORETA for the EEG fast spectral bands were evaluated in 42 teenagers with CD, 25 with and 17 without psychopathic traits according to the Antisocial Process Screening Device. All adolescents were assessed using the DSM-IV-TR criteria. The EEG visual inspection characteristics and the use of frequency domain quantitative analysis techniques (narrow band spectral parameters) are described. QEEG analysis showed a pattern of beta activity excess on the bilateral frontal-temporal regions and decreases of alpha band power on the left central-temporal and right frontal-central-temporal regions in the psychopathic traits group. Current source density calculated at 17.18 Hz showed an increase within fronto-temporo-striatal regions in the psychopathic relative to the nonpsychopathic traits group. These findings indicate that QEEG analysis and techniques of source localization may reveal differences in brain electrical activity among teenagers with CD and psychopathic traits, which was not obvious to visual inspection. Taken together, these results suggest that abnormalities in a fronto-temporo-striatal network play a relevant role in the neurobiological basis of psychopathic behavior.


High speed electro-optical cavity dumping of mode-locked laser oscillators.
High speed electro-optical cavity dumping is demonstrated with diode-pumped mode-locked laser oscillators, namely a femtosecond Yb:glass and a picosecond Nd:YVO4 oscillator. Repetition frequencies exceeding 1MHz are obtained with pulse energies of more than 300nJ /1 microJ. Being compact and easy to operate light sources, these laser systems open up various scientific and industrial applications.


Applied tension treatment of vasovagal syncope during pregnancy.
Vasovagal syncope is a common clinical problem that is often difficult and expensive to diagnose and treat. Applied tension is a behavioral treatment approach that has been demonstrated to be efficacious for the treatment of vasovagal syncope associated with injection phobia. The present case study evaluated the treatment of vasovagal syncope in a 41-year-old pregnant patient with injection phobia. The treatment included the use of applied muscle tension to increase blood pressure and prevent syncope during graduated exposure to increasingly greater anxiety-provoking stimuli. After completion of the treatment, the patient was able to undergo a blood draw and other medical procedures involving exposure to needles, with significantly reduced anxiety and no episodes of syncope. Applied tension is an effective and relatively inexpensive treatment for patients with vasovagal syncope related to injection phobia and may hold promise as a treatment for other types of syncope.


Pathogenesis of noroviruses, emerging RNA viruses.
Human noroviruses in the family Caliciviridae are a major cause of epidemic gastroenteritis. They are responsible for at least 95% of viral outbreaks and over 50% of all outbreaks worldwide. Transmission of these highly infectious plus-stranded RNA viruses occurs primarily through contaminated food or water, but also through person-to-person contact and exposure to fomites. Norovirus infections are typically acute and self-limited. However, disease can be much more severe and prolonged in infants, elderly, and immunocompromised individuals. Norovirus outbreaks frequently occur in semi-closed communities such as nursing homes, military settings, schools, hospitals, cruise ships, and disaster relief situations. Noroviruses are classified as Category B biodefense agents because they are highly contagious, extremely stable in the environment, resistant to common disinfectants, and associated with debilitating illness. The number of reported norovirus outbreaks has risen sharply since 2002 suggesting the emergence of more infectious strains. There has also been increased recognition that noroviruses are important causes of childhood hospitalization. Moreover, noroviruses have recently been associated with multiple clinical outcomes other than gastroenteritis. It is unclear whether these new observations are due to improved norovirus diagnostics or to the emergence of more virulent norovirus strains. Regardless, it is clear that human noroviruses cause considerable morbidity worldwide, have significant economic impact, and are clinically important emerging pathogens. Despite the impact of human norovirus-induced disease and the potential for emergence of highly virulent strains, the pathogenic features of infection are not well understood due to the lack of a cell culture system and previous lack of animal models. This review summarizes the current understanding of norovirus pathogenesis from the histological to the molecular level, including contributions from new model systems.


[Current options for antifungal therapy of invasive candidiasis in intensive care units].
Candidemia and invasive candidiasis are the most frequent mycoses in critically ill patients in intensive care units. Recently, the number of systemic antifungal agents has increased, leading to improved treatment options. Yet these infections remain to be characterized by poor prognosis and high mortality rates. The most important predisposing factors are yeast colonization of the mucosa or skin and damage to the integrity of the host's natural barriers. Early diagnosis of invasive candidiasis is difficult, since its clinical manifestations are not characteristic and the laboratory techniques are time-consuming and not completely reliable. The currently available treatments comprise three groups of antifungals: triazoles, polyenes and echinocandins. For its effectiveness, low toxicity and reasonable price, fluconazole is the most widespread drug currently used to treat systemic yeast infections. However, despite high treatment costs, echinocandins are becoming the drug of choice. The advantages are a broad spectrum of species, safe administration to patients with kidney and liver damage, minimal drug interactions and fungicidal effects. Candidemia may often be positively influenced by replacing an intravenous catheter. Despite earlier controversy, the latest treatment strategies clearly recommend its removal. Although antifungal prophylaxis lowers the incidence of invasive candidiasis, it is considered to be useful only if targeted to high-risk groups of patients. Empirical treatment of febrile patients not responding to broad-spectrum antibiotic therapy is only effective in wards with a higher incidence of systemic candidiasis, in patients with risk factors and if other causes are reliably excluded.


Aerial-aquatic robots capable of crossing the air-water boundary and hitchhiking on surfaces.
Many real-world applications for robots-such as long-term aerial and underwater observation, cross-medium operations, and marine life surveys-require robots with the ability to move between the air-water boundary. Here, we describe an aerial-aquatic hitchhiking robot that is self-contained for flying, swimming, and attaching to surfaces in both air and water and that can seamlessly move between the two. We describe this robot's redundant, hydrostatically enhanced hitchhiking device, inspired by the morphology of a remora (Echeneis naucrates) disc, which works in both air and water. As with the biological remora disc, this device has separate lamellar compartments for redundant sealing, which enables the robot to achieve adhesion and hitchhike with only partial disc attachment. The self-contained, rotor-based aerial-aquatic robot, which has passively morphing propellers that unfold in the air and fold underwater, can cross the air-water boundary in 0.35 second. The robot can perform rapid attachment and detachment on challenging surfaces both in air and under water, including curved, rough, incomplete, and biofouling surfaces, and achieve long-duration adhesion with minimal oscillation. We also show that the robot can attach to and hitchhike on moving surfaces. In field tests, we show that the robot can record video in both media and move objects across the air/water boundary in a mountain stream and the ocean. We envision that this study can pave the way for future robots with autonomous biological detection, monitoring, and tracking capabilities in a wide variety of aerial-aquatic environments.


Effects of an intubating dose of succinylcholine and rocuronium on the larynx and diaphragm: an electromyographic study in humans.
Paralysis of the vocal cords is one objective of using relaxants to facilitate tracheal intubation. This study compares the neuromuscular blocking effect of succinylcholine and rocuronium on the larynx, the diaphragm, and the adductor pollicis muscle. Electromyographic response was used to compare the neuromuscular blocking effect of succinylcholine and rocuronium on the laryngeal adductor muscles, the diaphragm, and the adductor pollicis muscle. Sixteen patients undergoing elective surgery were anesthetized with propofol and fentanyl, and their tracheas were intubated without neuromuscular blocking agents. The recurrent laryngeal and phrenic nerves were stimulated at the neck. The electromyographic response was recorded from electrodes placed on the endotracheal tube and intercostally before and after administration of 1 mg/kg succinylcholine or 0.6 mg/kg rocuronium. The maximum effect was greater at the adductor pollicis (100 and 99%) than at the larynx (96 and 97%) and the diaphragm (94 and 96%) after administration of succinylcholine and rocuronium, respectively (P < or = 0.05). Onset time was not different between the larynx (58+/-10 s), the diaphragm (57+/-8 s), and the adductor pollicis (54+/-13 s), after succinylcholine (all mean +/- SD). After rocuronium, onset time was 124+/-39 s at the larynx, 130+/-44 s at the diaphragm, and 115+/-21 s at the adductor pollicis. After succinylcholine administration, time to 90% recovery was 8.3+/-3.2, 7.2+/-3.5, and 9.1+/-3.0 min at the larynx, the diaphragm, and the adductor pollicis, respectively. Time to 90% recovery after rocuronium administration was 34.9+/-7.6, 30.4+/-4.2, and 49.1+/-11.4 min at the larynx, the diaphragm, and the adductor pollicis, respectively. Neuromuscular blocking effect of muscle relaxants on the larynx can be measured noninvasively by electromyography. Although the larynx appears to be resistant to muscle relaxants, we could not demonstrate that its onset time differed from that of peripheral muscles.


Gender-dependent multiple cross-phenotype association of interferon lambda genetic variants with peripheral blood profiles in healthy individuals.
Type III interferons (IFN), also called as lambda IFNs (IFN-λs), are antiviral and immunomodulatory cytokines that are evolutionarily important in humans. Given their central roles in innate immunity, they could be influencing other aspects of human biology. This study aimed to examine the association of genetic variants that control the expression and/or activity of IFN-λ3 and IFN-λ4 with multiple phenotypes in blood profiles of healthy individuals. In a cohort of about 550 self-declared healthy individuals, after applying several exclusion criteria to determine their health status, we measured 30 blood parameters, including cellular, biochemical, and metabolic profiles. We genotyped them at rs12979860 and rs28416813 using competitive allele-specific PCR assays and tested their association with the blood profiles under dominant and recessive models for the minor allele. IFN-λ4 variants rs368234815 and rs117648444 were also genotyped or inferred. We saw no association in the combined cohort under either of the models for any of the phenotypes. When we stratified the cohort based on gender, we saw a significant association only in males with monocyte (p = 1 × 10-3 ) and SGOT (p = 7 × 10-3 ) levels under the dominant model and with uric acid levels (p = 0.01) under the recessive model. When we tested the IFN-λ4 activity modifying variant within groupings based on absence or presence of one or two copies of IFN-λ4 and on different activity levels of IFN-λ4, we found significant (p < 0.05) association with several phenotypes like monocyte, triglyceride, VLDL, ALP, and uric acid levels, only in males. All the above significant associations did not show any confounding when we tested for the same with up to ten different demographic and lifestyle variables. These results show that lambda interferons can have pleiotropic effects. However, gender seems to be an effect modifier, with males being more sensitive than females to the effect.


Destabilization effect of transition metal fluorides on sodium borohydride.
The effect of transition metal fluorides on the decomposition of NaBH4 has been investigated for NaBH4 ball milled with TiF3, MnF3 or FeF3. The compounds were examined by thermal programmed desorption with residual gas analysis, thermo gravimetric analysis and volumetric measurements using a Sieverts-type apparatus. The phase formation process during thermal decomposition was studied by in situ synchrotron radiation powder X-ray diffraction on the as-milled powders. NaBF4 was among the products in all mechano-chemical reactions. (11)B-NMR spectra analysis gave NaBF4 : NaBH4 ratios of 1 : 150 for Na-Ti, 1 : 40 for Na-Mn, and 1 : 10 for Na-Fe. Pure NaBH4 possessed a hydrogen release onset temperature of 430 °C. The hydrogen release in the NaBH4-MnF3 system began as low as 130 °C. FeF3 decreased the onset temperature to 161 °C and TiF3 to 200 °C. TiF3 reacted completely with NaBH4 below 320 °C. All the examined systems have negligible emissions of diborane species. H-sorption studies performed at selected temperatures above 300 °C exhibited relatively fast desorption kinetics. Partial hydrogen re-absorption was observed for the Na-Mn and Na-Fe samples.


Quaternary ammoniums and other preservatives' contribution in oxidative stress and apoptosis on Chang conjunctival cells.
To investigate some of the toxicity mechanisms of 10 preservatives currently used in ophthalmic solutions in vitro. A continuous human conjunctival cell line was treated with different concentrations of various preservatives for 15 minutes and for 15 minutes followed by 24 hours of cell recovery: three benzalkonium chlorides (BACs) with different hydrocarbon chain length, benzododecinium bromide (BOB), cetrimide (Cet), phenylmercuric nitrate (PM), thimerosal (thi), methyl parahydroxybenzoate (MPHB), chlorobutanol (clb), and EDTA. An inhibition study was then conducted using a 1-hour vitamin E pretreatment followed by a 15-minute BAC treatment. Membrane integrity was assessed using a neutral red test and chromatin condensation with a Hoechst 33342 test. Reactive oxygen species were measured using dichlorofluorescein diacetate test for H2O2 production and hydroethidine test for O2.- production. These tests were performed using microplate cold light cytofluorometry. Cell size and DNA content were also analyzed using flow cytometry. Confocal microscopy was used to explore morphologic changes. A significant decrease of membrane integrity with chromatin condensation was observed with all the quaternary ammoniums tested at concentrations of 0.005% and higher. The effect was amplified after 24 hours of cell recovery. The other preservatives tested did not decrease membrane integrity. H2O2 production was observed with all the preservatives, whereas O2.- production was significantly higher with the quaternary ammoniums at 0.005% and 0.01%, compared with the other preservatives. Flow cytometry results confirmed the cytotoxicity observed with cold light cytofluorometry. The quaternary ammoniums tested (BAC, BOB, and Cet) were the most cytotoxic preservatives in the current model. An apoptotic mechanism appeared to be present at low concentrations of quaternary ammoniums, whereas a necrotic process appeared at higher concentrations. Superoxide anions may play an important role in tissue damage induced by preservatives in ocular surface disorders.


MICA polymorphism in South American Indians.
We have studied the MICA alleles of 196 unrelated subjects from three South American Indian tribes (Toba, Wichi and Terena). They are members of isolated tribes located in the Gran Chaco area in northeastern Argentina and in Mato Grosso do Sul in South Central Brazil. Of 55 previously known alleles, nine were observed in South American Indians, compared with 16 that were found in North American Caucasians, suggesting a more restricted allelic distribution of MICA in these tribes. In South American Indians, MICA*00201 was the most frequent allele, with a gene frequency of 33% in Toba, 47% in Wichi and 44% in Terena. MICA*00201, MICA*027 (external domain sequence like MICA*008/TM allele A5) and MICA*010 accounted for more than 90% of all the MICA genes in South American Indians. In North American Caucasians, MICA*00801 (*008/A5.1) accounted for 42% of the genes and was the most common allele. We observed a high degree of linkage disequilibrium between certain alleles of MICA and of HLA-B in the South American Indian populations. Phylogenetic trees constructed using gene frequencies of the transmembrane short tandem repeats in the populations reported here, and in other populations taken from published reports, suggest that South American Indians are more closely related to Asians than to Europeans.


Efficacy and tolerability of the modified Atkins diet in children with drug-resistant genetic generalized epilepsy.
The ketogenic diet therapy is a time-tested and potent non-pharmacologic treatment for epilepsy. However, the study of the ketogenic diet in patients with genetic generalized epilepsy (GGE) is not widely established. The aim of this study was to evaluate the efficacy and tolerability of the modified Atkins diet, a variation of the ketogenic diets, as a treatment for drug-resistant GGE. A retrospective chart review was performed in patients with epilepsy treated with the modified Atkins diet at the University of Chicago from 2017 to 2020. For three months following diet initiation, participants were monitored for diet tolerability and effect on seizures. Response to the treatment was recorded by self-reporting patients and guardians. Thirteen patients with a diagnosis of drug-resistant GGE were identified. An average of 3.8 anti-seizure medications (ASMs) had been tried and 3.4 years had elapsed from seizure onset before dietary therapy was attempted. Patients were receiving a mean of 2.2 ASMs at the time of diet initiation. After undergoing dietary treatment for three months, 12/13 (92%) patients experienced a greater than 50% reduction in seizure frequency, 6/13 (46%) patients became seizure-free, and 7/13 (54%) were able to discontinue at least one ASM. All patients completed at least three months of dietary therapy with an average duration of 9.3 months at the time of report. One patient reported side effects of fatigue which may be attributed to the diet. The modified Atkins diet has shown to be an effective and well-tolerated treatment for children with drug-resistant GGE. The diet provides the additional benefit of aiding to discontinue ASMs and, therefore, minimize the side effects from polypharmacy. Given these results, it seems reasonable to consider the modified Atkins diet as an alternative and possibly earlier treatment option for patients with drug-resistant GGE.


Occupation and risk of uveal melanoma. An exploratory study.
Little is known about the cause of uveal melanoma, the most common primary intraocular malignant lesion in adults. This population-based case-control study evaluated occupational exposures. One hundred ninety-seven newly diagnosed cases of uveal melanoma participated. Approximately two control subjects matched for age, sex, and telephone exchange area were selected for each case by random-digit dialing. Data were collected by a structured telephone interview. Two systems of occupational coding were used: an occupation-exposure linkage system and the Bureau of Census data. Conditional logistic regression for matched studies was used to examine various occupational exposures while controlling for previously reported potential confounders and for family income. The occupation-exposure matrix was used to define clusters of cases exposed to a particular chemical group from various occupations. Odds ratios were elevated for agriculture and farming work for both industry and occupation; this was consistent across both classification systems. Elevated odds ratios also were found for occupations involving machine operations, fabrication, assembling, equipment cleaning, and exposure to metal industries. Exposure to alkylating agents and phenols was associated with a lower risk compared with all other exposures. Several self-reported exposures also were studied. Exposures associated with elevated odds ratios were inks, insecticides, gases, radioactive substances, polybromated biphenyls, and chemical solvents. This exploratory study suggests various occupational associations for uveal melanoma and areas for future research.


Developing and Evaluating a Mobile Foot Care Application for Persons With Diabetes Mellitus: A Randomized Pilot Study.
Ulceration of the foot is a major complication of diabetes mellitus, and optimal self-care may help prevent its development. Research suggests that mobile applications (apps) may affect behavioral change. The purpose of this study was to develop the Mobile Diabetic Foot Personal Care System (m-DAKBAS) and evaluate its effectiveness for patients with diabetes. During Phase 1, a mobile app that included communication features, remote patient monitoring, and information was developed and pilot-tested among 10 patients. The Phase 2 study, conducted from June 2017 to April 2018, used a 2-group, pre-test/post-test design to evaluate the effect of the app on patients' knowledge, behavior, and self-efficacy scores when used for 6 months. Both the experimental (app) and control groups participated in 1 education session at the start of the study. Of 106 patients who enrolled, 88 completed the study (44 in the experimental group and 44 in the control group). Only 6 patients had received education about foot care previously. The average age of all participants was 51.63 years (SD = 8.08). There were significantly more women in the experimental group than in the control group (65% vs. 45.5%; P = 0.5). Each participant used the app for 24 weeks, and the data entry rate was 72.9%. Throughout the study, participants had 1977 data entries (blood glucose and foot observation) in total. Differences between pre- and post-intervention test scores were significantly higher for knowledge, behavior, and self-efficacy in both groups, but the difference was greater in the experimental group (P < .05). Only post-test knowledge scores were significantly higher in the experimental compared with the control group (P < .05). Compared to the start of the study, the proportion of participants with cracked/dry skin and inappropriate footwear was significantly lower in the experimental group but not in the control group. In this study, education and follow-up via the mobile app and verbal-only instruction increased the knowledge, behavior, and self-efficacy scores of patients in both groups. Post-study knowledge scores were significantly higher in the experimental group than in the control group. Patient education remains a crucial component of optimal care, and further development, refinement, and testing of mobile applications to improve self-efficacy and reduce the risk of diabetic foot are warranted.


Effect of template on generating a standard curve for absolute quantification of an RNA virus by real-time reverse transcriptase-polymerase chain reaction.
The effect of different templates on generating standard curves that are needed for the absolute quantification of an RNA virus by real-time reverse transcriptase-PCR (RT-PCR) was evaluated. We used infectious pancreatic necrosis virus (IPNV), a major viral pathogen of wild and cultured salmon, as a RNA virus example for the study. A dilution series of four different templates representing the IPNV protease gene (two in vitro transcribed RNAs of 100 bases and 500 bases in length, a plasmid DNA and a DNA oligo) were used as template to produce standard curves to quantify IPNV load in rainbow trout. The slope, the goodness of fit (r(2)), and the efficiency (e) of PCR were statistically equivalent irrespective of the nature of template used in the PCR. Using a factorial ANOVA, no significant difference in IPNV copy number was observed using the four different standard curves for absolute quantification of IPNV in experimentally-challenged rainbow trout. However, when IPNV transcript abundance was less than 100 copies per reaction and when the template size was bigger than the amplicon size amplification was more variable. The data suggests that the size of the template used to generate standard curve should be very similar to the size of the amplicon. A synthetic DNA oligo template would be optimal for this purpose as it can be custom made and only requires the sequence information for its synthesis. However, if the standard curve is generated with template copy number in excess of 100 copies per reaction, the nature of the template has no effect on the standard curve, and therefore, the cheaper template would be the preferred choice of template over the other more expensive options.


Aboriginal children and family connections to primary health care whilst homeless and in high housing mobility: observations from a Nurse Practitioner-led service.
This article documents the impact of a Nurse Practitioner-led primary health service for disadvantaged children living in housing instability or homelessness. It identifies that First Nations children miss out on essential primary care, particularly immunisation, but have less severe health conditions than non-First Nations children living in housing insecurity. Health services for homeless populations focus on the 11% of rough sleepers, little is done for the 22% of children in Australia living in housing instability; many of whom are from First Nations families. Little is known of the health status of these children or their connections to appropriate primary health care. This research implemented an innovative model of extended health care delivery, embedding a Nurse Practitioner in a homeless service to work with families providing health assessments and referrals, using clinically validated assessment tools. This article reports on proof of concept findings on the service that measured immunisation rates, developmental, medical, dental and mental health needs of children, particularly First Nations children, using a three-point severity level scale with Level 3 being the most severe and in need of immediate referral to a specialist medical service. Forty-three children were referred by the service to the Nurse Practitioner over a 6-month period, with nine identifying as First Nations children. Differences in severity levels between First Nations/non-First Nations children were Level 1, First Nations/non-First Nations 0/15%; Level 2, 10/17%; and Level 3, 45/29%. Forty-five percent of First Nations children had no health problems, as compared to 29% on non-First Nations children. Immunisation rates were low for both cohorts. No First Nations child was immunised and only 9% of the non-First Nations children. While numbers for both cohorts are too low for valid statistical analysis, the lower levels of severity for First Nations children suggest stronger extended family support and the positive impact of cultural norms of reciprocity.


Multinuclear NMR characterisation and dermal delivery of fluorinated drugs in soybean-microemulsion systems.
The present study evaluated the effect of different commercially available soybean lecithins in microemulsion systems in terms of microstructure transformation, physicochemical properties and transport of selected entrapped fluorinated drugs through skin. Physicochemical characterisations by particle size and polydispersity index (PDI) measurements were performed and a direct correlation with NMR self-diffusion coefficients of the individual components was found. An increase of lysophosphatidylcholine (LPC), phosphatidylethanolamine (PE) and lysophosphatidylethanolamine (LPE) in the phospholipid mixtures increased the mean particle sizes and PDI. Bicontinous microemulsion structures were proven by 1H and 31P NMR in the placebo microemulsions. Reasonable permeation of the lipophilic drugs of all microemulsions systems was confirmed in standard diffusion studies using porcine skin. This could be due to the incorporation of the drugs in the surfactant structure of the lecithin based bicontinous micro textures, as proven by 19F NMR self-diffusion studies.


Insulin-like growth factor binding proteins initiate cell death and extracellular matrix remodeling in the mammary gland.
We have demonstrated that insulin-like growth factor binding protein-5 (IGFBP-5) production by mammary epithelial cells increases dramatically during forced involution of the mammary gland in rats, mice and pigs. We proposed that growth hormone (GH) increases the survival factor IGF-I, whilst prolactin (PRL) enhances the effects of GH by decreasing the concentration of IGFBP-5, which would otherwise inhibit the actions of IGFs. To demonstrate a causal relationship between IGFBP-5 and cell death, we created transgenic mice expressing IGFBP-5, specifically, in the mammary gland. DNA content in the mammary glands of transgenic mice was decreased as early as day 10 of pregnancy. Mammary cell number and milk synthesis were both decreased by approximately 50% during the first 10 days of lactation. The concentrations of the pro-apoptotic molecule caspase-3 was increased in transgenic animals whilst the concentrations of two pro-survival molecules Bcl-2 and Bcl-x were both decreased. In order to examine whether IGFBP-5 acts by inhibiting the survival effect of IGF-I, we examined IGF receptor- and Akt-phoshorylation and showed that both were inhibited. These studies also indicated that the effects of IGFBP-5 could be mediated in part by IGF-independent effects involving potential interactions with components of the extracellular matrix involved in tissue remodeling, such as components of the plasminogen system, and the matrix metallo-proteinases (MMPs). Mammary development was normalised in transgenic mice by R3-IGF-I, an analogue of IGF-I which binds weakly to IGFBPs, although milk production was only partially restored. In contrast, treatment with prolactin was able to inhibit early involutionary processes in normal mice but was unable to prevent this in mice over-expressing IGFBP-5, although it was able to inhibit activation of MMPs. Thus, IGFBP-5 can simultaneously inhibit IGF action and activate the plasminogen system thereby coordinating cell death and tissue remodeling processes. The ability to separate these properties, using mutant IGFBPs, is currently under investigation.


Abnormal pelvic morphology and high cervical length are responsible for high-risk pregnancies in women displaying achondroplasia.
Pregnancies of women displaying achondroplasia are at high risk of adverse events. Early sonographic assessment of affected women can indicate an unusually long cervical length. It is the consequence of pathological anatomy of the pelvis. Thus, there is a foreseeable dystocia owing to cephalopelvic disproportion. Furthermore, this situation could also complicate cervical ripening prior to fetal extraction.