Characterization of the Haemophilus influenzae tehB gene and its role in virulence.
The Haemophilus influenzae ORF designated HI1275 in the Rd KW20 genomic sequence encodes a putative S-adenosyl methyltransferase with significant similarity to tellurite-resistance determinants (tehB) in other species. While the H. influenzae tehB can complement an Escherichia coli tehB mutation, thus restoring tellurite resistance, its role in H. influenzae is unknown. In a previous study defining the iron and haem modulon of H. influenzae, we showed that transcription of this gene in H. influenzae Rd KW20 increases during growth in iron- and haem-restricted media. Since iron and haem uptake genes, and other known virulence factors, constitute the majority of the iron- and haem-regulated gene set, we postulated that tehB may play a role in nutrient acquisition and/or the virulence of H. influenzae. A tehB mutant was constructed in the H. influenzae type b strain 10810 and was evaluated for growth defects in various supplemented media, as well as for its ability to cause infection in rat models of infection. Deletion of tehB leads to an increase in sensitivity both to tellurite and to the oxidizing agents cumene hydroperoxide, tert-butyl hydroperoxide and hydrogen peroxide. The tehB mutant additionally showed a significantly reduced ability to utilize free haem as well as several haem-containing moieties including haem-human serum albumin, haemoglobin and haemoglobin-haptoglobin. Examination of the regulation kinetics indicated that transcription of tehB was independent of both tellurite exposure and oxidative stress. Paired comparisons of the tehB mutant and the wild-type H. influenzae strain 10810 showed that tehB is required for wild-type levels of infection in rat models of H. influenzae invasive disease. To our knowledge this is the first report of a role for tehB in virulence in any bacterial species. These data demonstrate that H. influenzae tehB plays a role in both resistance to oxidative damage and haem uptake/utilization, protects H. influenzae from tellurite exposure, and is important for virulence of this organism in an animal model of invasive disease.


A mathematical model of the metastatic bottleneck predicts patient outcome and response to cancer treatment.
Metastases are the main reason for cancer-related deaths. Initiation of metastases, where newly seeded tumor cells expand into colonies, presents a tremendous bottleneck to metastasis formation. Despite its importance, a quantitative description of metastasis initiation and its clinical implications is lacking. Here, we set theoretical grounds for the metastatic bottleneck with a simple stochastic model. The model assumes that the proliferation-to-death rate ratio for the initiating metastatic cells increases when they are surrounded by more of their kind. For a total of 159,191 patients across 13 cancer types, we found that a single cell has an extremely low median probability of successful seeding of the order of 10-8. With increasing colony size, a sharp transition from very unlikely to very likely successful metastasis initiation occurs. The median metastatic bottleneck, defined as the critical colony size that marks this transition, was between 10 and 21 cells. We derived the probability of metastasis occurrence and patient outcome based on primary tumor size at diagnosis and tumor type. The model predicts that the efficacy of patient treatment depends on the primary tumor size but even more so on the severity of the metastatic bottleneck, which is estimated to largely vary between patients. We find that medical interventions aiming at tightening the bottleneck, such as immunotherapy, can be much more efficient than therapies that decrease overall tumor burden, such as chemotherapy.


Capillary basement membrane width in diabetic children.
The effect of juvenile onset diabetes mellitus on quadriceps muscle capillary basement membrane (QCBM) width has been examined by the electron microscopic morphometric method previously developed in this laboratory. The results demonstrate that in this age group QCBM thickening is strongly related to the age of the diabetic subject. As a result, in contrast to the almost constant thickening of QCBM that has consistently been documented in diabetic adults, QCBM hypertrophy is present in only 40 per cent of children with diabetes mellitus. As was previously shown to be the case in adults, in children, too, QCBM thickening is unrelated to the duration of the diabetes. Finally, the finding that QCBM hypertrophy is present at the time of acute onset of juvenile diabetes mellitus in 30 per cent of children, coupled with the fact that this lesion is not affected by duration of hyperglycemia, strongly supports our previous conclusion that diabetic microangiopathy is independent of the hyperglycemia of this disease. On the other hand, barring the possibility that microangiopathy in the pancreas precedes that in muscle, these results represent evidence against the suggestion that basement membrane hypertrophy represents the primary lesion of the diabetic syndrome.


The Lipid Energy Model: Reimagining Lipoprotein Function in the Context of Carbohydrate-Restricted Diets.
When lean people adopt carbohydrate-restricted diets (CRDs), they may develop a lipid profile consisting of elevated LDL-cholesterol (LDL-C) and HDL-cholesterol (HDL-C) with low triglycerides (TGs). The magnitude of this lipid profile correlates with BMI such that those with lower BMI exhibit larger increases in both LDL-C and HDL-C. The inverse association between BMI and LDL-C and HDL-C change on CRD contributed to the discovery of a subset of individuals-termed Lean Mass Hyper-Responders (LMHR)-who, despite normal pre-diet LDL-C, as compared to non-LMHR (mean levels of 148 and 145 mg/dL, respectively), exhibited a pronounced hyperlipidemic response to a CRD, with mean LDL-C and HDL-C levels increasing to 320 and 99 mg/dL, respectively, in the context of mean TG of 47 mg/dL. In some LMHR, LDL-C levels may be in excess of 500 mg/dL, again, with relatively normal pre-diet LDL-C and absent of genetic findings indicative of familial hypercholesterolemia in those who have been tested. The Lipid Energy Model (LEM) attempts to explain this metabolic phenomenon by positing that, with carbohydrate restriction in lean persons, the increased dependence on fat as a metabolic substrate drives increased hepatic secretion and peripheral uptake of TG contained within very low-density lipoproteins (VLDL) by lipoprotein lipase, resulting in marked elevations of LDL-C and HDL-C, and low TG. Herein, we review the core features of the LEM. We review several existing lines of evidence supporting the model and suggest ways to test the model's predictions.


Equivalence of two healthcare costing methods: bottom-up and top-down.
This paper compares two quite different approaches to estimating costs: a 'bottom-up' approach, represented by the US Department of Veterans Affairs' (VA) Decision Support System that uses local costs of specific inputs; and a 'top-down' approach, represented by the costing system created by the VA Health Economics Resource Center, which assigns the VA national healthcare budget to specific products using various weighting systems. Total annual costs per patient plus the cost for specific services (e.g. clinic visit, radiograph, laboratory, inpatient admission) were compared using scatterplots, correlations, mean difference, and standard deviation of individual differences. Analysis are based upon 2001 costs for 14 915 patients at 72 facilities. Correlations ranged from 0.24 for the cost of outpatient encounters to 0.77 for the cost of inpatient admissions, and 0.85 for total annual cost. The mean difference between costing methods was $707 ($4168 versus $3461) for total annual cost. The standard deviation of the individual differences was $5934. Overall, the agreement between the two costing systems varied by the specific cost being measured and increased with aggregation. Administrators and researchers conducting cost analyses need to carefully consider the purpose, methods, characteristics, strengths, and weaknesses when selecting a method for assessing cost.


Post-eclosion temperature effects on insect cuticular hydrocarbon profiles.
The insect cuticle is the interface between internal homeostasis and the often harsh external environment. Cuticular hydrocarbons (CHCs) are key constituents of this hard cuticle and are associated with a variety of functions including stress response and communication. CHC production and deposition on the insect cuticle vary among natural populations and are affected by developmental temperature; however, little is known about CHC plasticity in response to the environment experienced following eclosion, during which time the insect cuticle undergoes several crucial changes. We targeted this crucial to important phase and studied post-eclosion temperature effects on CHC profiles in two natural populations of Drosophila melanogaster. A forty-eight hour post-eclosion exposure to three different temperatures (18, 25, and 30°C) significantly affected CHCs in both ancestral African and more recently derived North American populations of D. melanogaster. A clear shift from shorter to longer CHCs chain length was observed with increasing temperature, and the effects of post-eclosion temperature varied across populations and between sexes. The quantitative differences in CHCs were associated with variation in desiccation tolerance among populations. Surprisingly, we did not detect any significant differences in water loss rate between African and North American populations. Overall, our results demonstrate strong genetic and plasticity effects in CHC profiles in response to environmental temperatures experienced at the adult stage as well as associations with desiccation tolerance, which is crucial in understanding holometabolan responses to stress.


Acute vitreoretinal trauma and inflammation after traumatic brain injury in mice.
Limited attention has been given to ocular injuries associated with traumatic brain injury (TBI). The retina is an extension of the central nervous system and evaluation of ocular damage may offer a less-invasive approach to gauge TBI severity and response to treatment. We aim to characterize acute changes in the mouse eye after exposure to two different models of TBI to assess the utility of eye damage as a surrogate to brain injury. A model of blast TBI (bTBI) using a shock tube was compared to a lateral fluid percussion injury model (LFPI) using fluid pressure applied directly to the brain. Whole eyes were collected from mice 3 days post LFPI and 24 days post bTBI and were evaluated histologically using a hematoxylin and eosin stain. bTBI mice showed evidence of vitreous detachment in the posterior chamber in addition to vitreous hemorrhage with inflammatory cells. Subretinal hemorrhage, photoreceptor degeneration, and decreased cellularity in the retinal ganglion cell layer was also seen in bTBI mice. In contrast, eyes of LFPI mice showed evidence of anterior uveitis and subcapsular cataracts. We demonstrated that variations in the type of TBI can result in drastically different phenotypic changes within the eye. As such, molecular and phenotypic changes in the eye following TBI may provide valuable information regarding the mechanism, severity, and ongoing pathophysiology of brain injury. Because vitreous samples are easily obtained, molecular changes within the eye could be utilized as biomarkers of TBI in human patients.


Matrix metalloprotease-2 and -9 concentration and activity in serum and culture medium samples: a methodological reappraisal.
Matrix metalloproteases (MMPs) play an important role in cardiovascular remodeling by degrading the extracellular matrix. The aim of this study was to compare two different methods for MMP-2 and MMP-9 concentration and activity determination. MMP-2 and -9 levels were measured by immunometric and enzymatic assays to determine total and active levels. The two procedures differ in assay principle and in the extent of cross-reactions with interfering substances present in biological samples. Both human serum and culture medium from an ex vivo human model of intimal hyperplasia were checked. All methods were able to detect MMP-2 and -9 with similar levels of sensitivity, reproducibility and accuracy, and furnished positively related results, although significantly different, in both types of sample. Both systems were able to detect changes in MMP production such as the time-course of MMP-2 and -9 release by cultured saphenous vein associated with intima hyperplasia progression. Our data indicate that different values for MMP concentrations can be obtained using different analytical methods, even if they are intrinsically reliable. This suggests that methodological differences should be taken into account when comparing MMP results from different studies.


Bibliography of research utilizing the vividness of visual imagery questionnaire.
The Vividness of Visual Imagery Questionnaire (VVIQ) is receiving increasing use as a self-report measure of mental, visual imagery vividness. This bibliography contains lists of the publications and dissertations which report investigations of experimental, correlational, and factor analytical research. It also includes publications in which research utilizing the VVIQ is reviewed, critiqued, and discussed. The bibliography provides a resource for investigators.


Patient-reported complications associated with Campylobacter jejuni infection.
This study aimed to investigate the occurrence of complications, especially musculoskeletal symptoms, after sporadic Campylobacter jejuni enteritis of domestic origin in Finland. This multi-centre cross-sectional study was conducted during a seasonal peak in 2002. Questionnaires were sent to Campylobacter-positive patients, representing different geographical areas, 2 months after collection of positive stool samples. Medical records were viewed in several cases. Besides antimicrobial susceptibility testing C. jejuni isolates were serotyped. A total of 235 patients (58%) returned the questionnaire and 201 C. jejuni-positive patients were finally included in the study. Musculoskeletal symptoms associated with C. jejuni enteritis were frequent (39%); joint pain was most commonly reported (81%). The incidence of reactive arthritis was 4% and that of Achilles enthesopathy and/or heel pain was 9%. Stomach ache during enteritis was associated with the later development of joint pain. Antimicrobial treatment was common but did not prevent complications.


Effect of concurrent experimentally induced bovine respiratory syncytial virus and bovine viral diarrhea virus infection on respiratory tract and enteric diseases in calves.
To compare experimentally induced concurrent bovine viral diarrhea virus (BVDV) and bovine respiratory syncytial virus (BRSV) infection with single virus infection. 9- to 12-month-old calves. Calves were allotted to 4 groups: 1, mock-infected control (n = 3); 2, BRSV infected (5); 3, BVDV infected (5); and 4, concurrent BRSV and BVDV infected (5). Total and differential WBC counting was done. Concentration and duration of BVDV in nasal secretions and serum, and duration of BRSV in nasal secretions were determined. Concentration of BVDV in various tissues was determined, and isolation of BRSV from lung tissue was attempted. Histologic examination and immunohistochemical analysis were done to detect lesions and distribution of viral antigens, respectively. Calves with concurrent infection developed more severe clinical signs of disease (fever and diarrhea), leukopenia, and more severe lesions. They also shed virus from nasal secretions in greater concentration and for longer duration, and BRSV was isolated from their lungs. Calves with concurrent infection also had more extensive lung lesions. Alimentary epithelial necrosis and severe lymphoid depletion were associated with BVDV infection in calves with or without concurrent BRSV infection. BVDV antigen in lymphatic tissue was detected in stromal cells only. Concurrent infection with BRSV and BVDV resulted in more severe respiratory tract and enteric disease than did infection with either virus alone, possibly indicating synergistic effect between the viruses. BVDV's role in causing respiratory tract disease is attributable, indirectly, to effects on the host's immune system, not to infection of the lungs.


Conjugative transfer genes in staphylococcal isolates from the United States.
Staphylococcus aureus and coagulase-negative staphylococcal isolates from various geographic areas in the United States were examined by using a conjugative transfer gene DNA probe in dot-blot hybridization assays. Of 175 S. aureus isolates, 47 (27%) hybridized with the probe, while 24 of 208 (11.5%) coagulase-negative staphylococci hybridized. However, among methicillin-resistant S. aureus 52% (45 of 89) were probe positive while only 2% (2 of 86) of methicillin-susceptible S. aureus were probe positive. In contrast, 12.5% (22 of 176) of methicillin-resistant and 6% (2 of 32) of methicillin-susceptible coagulase-negative staphylococci contained transfer genes. All but one of the staphylococci containing transfer genes were resistant to gentamicin; 91.5% of S. aureus and 65% of coagulase-negative staphylococci containing transfer genes transferred gentamicin resistance to a S. aureus recipient. Of the 12 isolates that hybridized with the probe but did not transfer resistance, 10 (6 coagulase-negative staphylococci and 4 S. aureus) carried both gentamicin resistance and conjugative transfer genes on the same plasmid. Of these 10, 6 contained plasmid target fragments of sizes different from that of the probe, suggesting additions or deletions of DNA essential for transfer, while in 4 no such alterations could be detected. In two coagulase-negative staphylococci the entire transfer region was apparently integrated into the chromosome. Thus, staphylococci carrying conjugative transfer genes are widely disseminated in the United States and are usually found in multiresistant isolates on plasmids that also encode gentamicin resistance.


Can natural language processing help differentiate inflammatory intestinal diseases in China? Models applying random forest and convolutional neural network approaches.
Differentiating between ulcerative colitis (UC), Crohn's disease (CD) and intestinal tuberculosis (ITB) using endoscopy is challenging. We aimed to realize automatic differential diagnosis among these diseases through machine learning algorithms. A total of 6399 consecutive patients (5128 UC, 875 CD and 396 ITB) who had undergone colonoscopy examinations in the Peking Union Medical College Hospital from January 2008 to November 2018 were enrolled. The input was the description of the endoscopic image in the form of free text. Word segmentation and key word filtering were conducted as data preprocessing. Random forest (RF) and convolutional neural network (CNN) approaches were applied to different disease entities. Three two-class classifiers (UC and CD, UC and ITB, and CD and ITB) and a three-class classifier (UC, CD and ITB) were built. The classifiers built in this research performed well, and the CNN had better performance in general. The RF sensitivities/specificities of UC-CD, UC-ITB, and CD-ITB were 0.89/0.84, 0.83/0.82, and 0.72/0.77, respectively, while the values for the CNN of CD-ITB were 0.90/0.77. The precisions/recalls of UC-CD-ITB when employing RF were 0.97/0.97, 0.65/0.53, and 0.68/0.76, respectively, and when employing the CNN were 0.99/0.97, 0.87/0.83, and 0.52/0.81, respectively. Classifiers built by RF and CNN approaches had excellent performance when classifying UC with CD or ITB. For the differentiation of CD and ITB, high specificity and sensitivity were achieved as well. Artificial intelligence through machine learning is very promising in helping unexperienced endoscopists differentiate inflammatory intestinal diseases. The abstract of this article has won the first prize of the Young Investigator Award during the Asian Pacific Digestive Week (APDW) 2019 held in Kolkata, India.


Systematic review of ventricular peritoneal shunt and percutaneous endoscopic gastrostomy: a safe combination.
OBJECTIVE Various international and national gastrointestinal guidelines take different positions on whether ventriculoperitoneal shunt (VPS) insertion is a contraindication to percutaneous endoscopic gastrostomy (PEG). The objective of this meta-analysis was to try to answer the question of whether VPS insertion is a contraindication to PEG. METHODS A systematic review of the literature was performed according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) criteria. Electronic databases PubMed and Embase were searched using variations of the terms "ventriculo-peritoneal shunt" and "percutaneous (endoscopic) gastrostomy." This search resulted in 70 studies, 9 of which were relevant. These were cross-referenced, and 1 additional study was found, resulting in 10 studies in this systematic review. RESULTS The 10 relevant studies in adult cohorts included 208 patients. All studies save one were retrospective and, in general, poor quality. Among the studies with relevant data, there were 26 (12.5% of 208 cases) VPS infections and 4 (4.4% of 90 cases) VPSs that malfunctioned. In 137 patients the VPS had been placed before the PEG tube, with a VPS infection rate of 4.4%. More VPS infections occurred among the 55 patients who first had a PEG and a subsequent VPS (21.8%) and in the 16 patients who had simultaneous PEG tube and VPS placement (50%). The heterogeneity of the studies in this analysis prohibited statistical comparisons of the timing of VPS and PEG tube placement. CONCLUSIONS This systematic review indicated that VPS placement in combination with a PEG has a high but acceptable VPS complication rate. Therefore, VPS insertion should not be considered a contraindication to the placement of a PEG tube. Preferably, a PEG tube should be placed after the VPS. Waiting 7-10 days between VPS insertion and a PEG seems reasonable, but this could not be corroborated in this review.


Impact of antigenic exposures and role of molecular blood grouping in enhancing transfusion safety in chronically transfused thalassemics.
Red cell alloimmunization is an acknowledged complication of blood transfusion. Current transfusion practices for thalassemia do not cater to this risk. Serological phenotyping is usually not reliable in these cases unless performed before the first transfusion. Under such circumstances, molecular blood grouping is an effective alternative. To perform molecular blood group genotyping in chronically transfused thalassemia patients and assess the risk of antigenic exposure and incidence of alloimmunization with current transfusion protocols. Molecular blood group genotyping was performed for 47 chronically transfused thalassemia patients. Their 1-year transfusion records were retrieved to assess the antigenic exposure and the frequency thereof. Of 47 patients, 6 were already alloimmunized (3 with anti-E and 3 with anti-K) and were receiving the corresponding antigen negative units. We observed that random selection of ABO and Rh D matched units resulted in 57.7% ±8.26% chance of Rh and Kell phenotype matching also. Forty-four patients had received one or more antigenic exposures at least once. The 6 already alloimmunized patients were further exposed to antigens other than the ones they were immunized to. During the study period, only one patient developed an alloantibody, anti-E with exposure to antigens C (92%) and/or E (32%) at each transfusion. Several factors apart from mere antigen exposure may influence the development of alloimmunization as most of our patients received antigenic exposures but not alloimmunized. Our data provide an impetus for future large-scale studies to understand the development of alloimmunization in such patients.


[The spatial organization of the afferent projections in the canine nucleus accumbens].
Using the retrograde axonal marker transport it was shown that in dog nucleus accumbens medial and lateral segments receive similar projections. Cortical ones receive projections from mesocortical (prelimbic, infralimbic, orbital, insular and cingular) cortical fields, whereas subcortical receive projections from ventral tegmental field and basal nucleus of amygdala. The differences between projections of nucleus accumbens studied are that apart from the above-mentioned common afferents medial segment receives projections from allocortical (entorhinal, periamygdalar) fields and subicular portion of hippocampal formation while lateral segment receives projections from all dopaminergic portions of substantia nigra.


A novel GIS-MCDA-based spatial habitat suitability model for Posidonia oceanica in the Mediterranean.
Seagrasses are extremely productive flowering plants that produce oxygen by photosynthesis in the marine environment. They are globally in decline and can become endangered due to human activities and natural hazards. In order to maintain seagrass biodiversity, suitable habitats for this species must be determined and marine protected areas must be established. Recent technology allows acoustic systems to collect accurate high resolution data of the seafloor. Additionally, cost-effective optical satellite images, which provide wide coverage, have been used in various benthic studies. In this study, a habitat suitability model was developed using acoustic and optical data for Posidonia oceanica in Gulluk Bay, Turkey, SE Mediterranean, by applying the geographic information system-multi-criteria decision analysis and remote sensing techniques. Various criteria, namely, depth, sheltered area, slope, sediment yield, and topographic position index, were weighted using the analytic hierarchy process method. The model was able to identify suitable habitats for seagrass with 76% accuracy. The proposed model in the study allows fast, temporal, cost-effective, and sustainable production of seagrass habitat maps.


Biglycan expression in the melanoma microenvironment promotes invasiveness via increased tissue stiffness inducing integrin-β1 expression.
Novel targeted and immunotherapeutic approaches have revolutionized the treatment of metastatic melanoma. A better understanding of the melanoma-microenvironment, in particular the interaction of cells with extracellular matrix molecules, may help to further improve these new therapeutic strategies.We observed that the extracellular matrix molecule biglycan (Bgn) was expressed in certain human melanoma cells and primary fibroblasts when evaluated by microarray-based gene expression analysis. Bgn expression in the melanoma tissues correlated with low overall-survival and low progression-free-survival in patients. To understand the functional role of Bgn we used gene-targeted mice lacking functional Bgn. Here we observed that melanoma growth, metastasis-formation and tumor-related death were reduced in Bgn-/- mice compared to Bgn+/+ mice. In vitro invasion of melanoma cells into organotypic-matrices derived from Bgn-/- fibroblasts was reduced compared to melanoma invasion into Bgn-proficient matrices. Tissue stiffness as determined by atomic-force-microscopy was reduced in Bgn-/- matrices. Isolation of melanoma cells and fibroblasts from the stiffer Bgn+/+ matrices revealed an increase in integrin-β1 expression compared to the Bgn-/- fibroblast matrices. Overexpression of integrin-β1 in B16-melanoma cells abolished the survival benefit seen in Bgn-/- mice. Consistent with the studies performed in mice, the abundance of Bgn-expression in human melanoma samples positively correlated with the expression of integrin-β1, which is in agreement with results from the organotypic invasion-assay and the in vivo mouse studies.This study describes a novel role for Bgn-related tissue stiffness in the melanoma-microenvironment via regulation of integrin-β1 expression by melanoma cells in both mice and humans.


New insights into the structure-spectrum relationship in S65T/H148D and E222Q/H148D green fluorescent protein mutants: a theoretical assessment.
The green fluorescent protein (GFP) variant S65T/H148D recovers the A-band fluorescence lost in the single mutant S65T, and it has been established that Asp148 is the alternate proton acceptor for the excited state proton transfer (ESPT). This mutant has been widely studied and presents unique spectroscopic properties, such as an ultrafast rise in the fluorescence (<50 fs). Also it exhibits a red-shift of the A absorption band of 20 nm with respect to wt-GFP's. The double mutant E222Q/H148D presents a very similar behaviour, at least within the experimental data available (which is scarcer than those of S65T/H148D). By means of dynamic theoretical studies we have been able to (1) reproduce and thoroughly analyse the red-shifted absorption spectra of both mutants and (2) predict the structure that the variant E222Q/H148D (for which there is no X-ray-resolved structure available) most probably adopts in water at room temperature. Our results deepen the understanding of the way GFP variants work and give some new insights into the rational design of fluorescent proteins and biological photosystems in general.


Marmosets in New World leishmaniasis research.
In leishmaniasis, the search for a suitable experimental animal model is highly recommended, especially in Leishmania (Viannia) braziliensis (L. (V) b) research. Diverse species of neotropical primates have increasingly been used as experimental hosts of Leishmania. This article describes aspects of parasite L(V)b and marmoset, Callithrix penicillata (Primates, Callithricidae), interaction and summarizes the biology of parasitism in this primate model. Results of recent studies on primary infections with several strains of L(V)b as well as parasitism evolution, clinical outcome patterns and immunoprotection experiments, are summarily discussed. The relevance of homologous reinfection experiments for vaccine development in leishmaniasis is discussed.


Construction of artificial methemoglobin reduction systems in Hb vesicles.
The hemoglobin vesicle (HbV) is a red cell substitute encapsulating purified concentrated Hb in a phospholipid vesicle. In order to suppress metHb formation for the long term maintenance of oxygen transporting capability in vivo, thiols (cysteine, Cys; homocysteine, Hcy) were studied as reductants of metHb. Hcy showed a suppressive effect on metHb formation, while Cys adversely accelerates metHb formation at the rate of twice the Hb solution without any reductants. The suppression of Cys-induced metHb formation by the addition of superoxide dismutase (SOD) and catalase indicated that Cys was easily oxidized by oxygen and simultaneously generated a large amount of active oxygens. The rate of metHb formation was influenced by PO2 and pH. Furthermore, the reducing systems (methylene blue (MB), NADH or ascorbic acid) were added to the outer aqueous phase of HbV, and the artificial reduction systems constructed through the bilayer membrane were evaluated.


History of foster care among homeless adults with mental illness in Vancouver, British Columbia: a precursor to trajectories of risk.
It is well documented that a disproportionate number of homeless adults have childhood histories of foster care placement(s). This study examines the relationship between foster care placement as a predictor of adult substance use disorders (including frequency, severity and type), mental illness, vocational functioning, service use and duration of homelessness among a sample of homeless adults with mental illness. We hypothesize that a history of foster care predicts earlier, more severe and more frequent substance use, multiple mental disorder diagnoses, discontinuous work history, and longer durations of homelessness. This study was conducted using baseline data from two randomized controlled trials in Vancouver, British Columbia for participants who responded to a series of questions pertaining to out-of-home care at 12 months follow-up (n = 442). Primary outcomes included current mental disorders; substance use including type, frequency and severity; physical health; duration of homelessness; vocational functioning; and service use. In multivariable regression models, a history of foster care placement independently predicted incomplete high school, duration of homelessness, discontinuous work history, less severe types of mental illness, multiple mental disorders, early initiation of drug and/or alcohol use, and daily drug use. This is the first Canadian study to investigate the relationship between a history of foster care and current substance use among homeless adults with mental illness, controlling for several other potential confounding factors. It is important to screen homeless youth who exit foster care for substance use, and to provide integrated treatment for concurrent disorders to homeless youth and adults who have both psychiatric and substance use problems. Both trials are registered with the International Standard Randomized Control Trial Number Register and were assigned ISRCTN57595077 (Vancouver At Home Study: Housing First plus assertive community treatment versus congregate housing plus supports versus treatment as usual) and ISRCTN66721740 (Vancouver At Home Study: Housing First plus intensive case management versus treatment as usual) on September 9, 2012.


Metallic contamination of food during preparation and storage: development of methods and some preliminary results.
A chemical procedure for studying trace metals leached from metallic cooking utensils and preserving cans used in the preparation and storage of food has been developed. The method consists in the destruction of the major part of organic matter with HNO3-vapour followed by a complete mineralization of residues with small amounts of HNO3 in Teflon bombs at 150-160 degrees C under a pressure of 3-12 kg/cm2, depending on the amount and composition of the samples. Subsequently, an ion-exchange step removes major components and concentrates the trace elements in a dilute HNO3-solution, suitable for analysis. The ion-exchange separation, which is performed with an automatic ion-exchange separator, is practically free from blank level problems, e.g., typically a mean of less than 2 per cent of the sample levels of the elements being determined. Preliminary results show that large amounts of aluminium are released from vessels to the water during boiling at the same pH-range which exist for most drinking water in Sweden.


The effects of sildenafil on human sperm function in healthy volunteers.
This double-blind, randomized, four-period, two-way crossover study was conducted to evaluate the acute effects of oral sildenafil (100-mg single dose) on sperm motility, count, density, morphology and vitality as well as ejaculate volume and viscosity in healthy male subjects. The concentrations of sildenafil and its primary circulating metabolite UK-103,320 were measured in ejaculate and compared with those in plasma. The study also included assessments of safety and tolerability. A total of 17 healthy male volunteers aged 19--34 years were randomized to receive a single 100-mg dose of sildenafil for two periods and a single dose of placebo for two periods, with each period separated by a minimum of 5--7 days. Sperm and ejaculate properties were evaluated from semen samples taken at screening and 1.5 h after dose. An additional semen sample was collected 4 h after dose, and drug and metabolite concentrations were measured in this sample and the sample taken 1.5 h after dose for comparison with plasma concentrations. Blood samples were collected before each dose and 0.25, 0.5, 1, 2, 3, 4 and 6 h after dose for measurement of sildenafil and metabolite concentrations. Sildenafil had no statistically significant effect on sperm motility, count or density; the percentage of abnormal sperm forms; or the percentage of living sperm. It also did not affect ejaculate volume or viscosity. All measures were within normal ranges. Sildenafil distributed into the semen rapidly, resulting in significant correlations between concentrations of sildenafil in the semen and total (R2=0.588) or free (R2=0.454) plasma concentrations (P<0.0001). Total semen concentrations of sildenafil were 18% of total plasma concentrations. UK-103,320 appeared to distribute more slowly from the plasma into the semen, resulting in a lack of correlation between semen and plasma concentrations. The amount of sildenafil and UK-103,320 in the ejaculate was small (< 2x10(-4)% of the administered dose at 1.5 h). Sildenafil was well tolerated; no patient withdrew from the study due to adverse events attributed to sildenafil. These results indicate that a single 100-mg oral dose of sildenafil does not have an adverse effect on sperm function or ejaculate quality.


MHC diversity, malaria and lifetime reproductive success in collared flycatchers.
Major histocompatibility complex (MHC) genes encode proteins involved in the recognition of parasite-derived antigens. Their extreme polymorphism is presumed to be driven by co-evolution with parasites. Host-parasite co-evolution was also hypothesized to optimize within-individual MHC diversity at the intermediate level. Here, we use unique data on lifetime reproductive success (LRS) of female collared flycatchers to test whether LRS is associated with within-individual MHC class II diversity. We also examined the association between MHC and infection with avian malaria. Using 454 sequencing, we found that individual flycatchers carry between 3 and 23 functional MHC class II B alleles. Predictions of the optimality hypothesis were not confirmed by our data as the prevalence of blood parasites decreased with functional MHC diversity. Furthermore, we did not find evidence for an association between MHC diversity and LRS.


[Significance of apolipoproteins A and B and the remaining lipid fractions as indicators of protein-calorie malnutrition in the elderly].
Effects of calorie/protein malnourishment have been studied on plasma concentrations of cholesterol, triglycerides, HDL-C, LDL-C and apolipoproteins A and B, in institutionalized elderly people, 53 males and 62 females, of whom 19 females and 12 males were malnourished. In malnourished patients, total cholesterol and LDL-C were significantly lower both in males and in females, but HDL-C was lower only in females. No significant differences in plasma triglycerides were found between the control and the malnourished groups. Apolipoprotein A showed no significant changes on malnourished males, but did show a significant lowering in malnourished females. On the opposite, apolipoprotein B was lower in males than in females. The lowering in cholesterol in malnourished patients leads us to think that this could be a early predictor of nutritional risk in the elderly.


Diagnostic criteria and adjudication process both determine published event-rates: the ACTION trial experience.
Few trials report event-adjudication procedures in detail. Using data from the ACTION (A Coronary disease Trial Investigating Outcome with Nifedipine GITS) study, we compared the impact on event-rates of an adjudication strategy based on systematic screening of all reported serious adverse events (SAEs) with a strategy based on investigator diagnoses. The final diagnosis was always made by a critical events committee (CEC) using standard criteria. ACTION randomized 7665 patients with stable angina to either nifedipine or placebo. Pre-specified events included acute or procedural myocardial infarction (MI), refractory angina, heart failure and debilitating stroke. Clinically related SAEs including in-hospital procedures were combined into episodes independent from the investigator diagnoses entered on SAE reports. All fatal episodes and those episodes suggestive of pre-specified events were adjudicated by the CEC. During follow-up, 17,081 episodes were reported in 5312 patients. The SAE descriptions ruled out the occurrence of a pre-specified event in 28%. The remaining 72% were adjudicated by the CEC and 616 cases of MI, 361 of refractory angina, 275 of heart failure and 190 of debilitating stroke were diagnosed (total=1442). Had adjudication by the CEC been limited to the 3924 episodes (2397 patients) that were fatal or for which the investigator had reported any of the diagnoses mentioned, 98 cases of MI, 35 of refractory angina, 81 of heart failure and 14 of debilitating stroke would have been missed (total=228). Both the diagnostic criteria used and the adjudication process determine event-rates and conclusions about treatment effects in clinical trials. Published trial reports should always state if event-adjudication was independent of the diagnoses of investigators, and if all events of interest were adjudicated or only the first one.


Histone deacetylase 6 acts upstream of DNA damage response activation to support the survival of glioblastoma cells.
DNA repair promotes the progression and recurrence of glioblastoma (GBM). However, there remain no effective therapies for targeting the DNA damage response and repair (DDR) pathway in the clinical setting. Thus, we aimed to conduct a comprehensive analysis of DDR genes in GBM specimens to understand the molecular mechanisms underlying treatment resistance. Herein, transcriptomic analysis of 177 well-defined DDR genes was performed with normal and GBM specimens (n = 137) from The Cancer Genome Atlas and further integrated with the expression profiling of histone deacetylase 6 (HDAC6) inhibition in temozolomide (TMZ)-resistant GBM cells and patient-derived tumor cells. The effects of HDAC6 inhibition on DDR signaling were examined both in vitro and intracranial mouse models. We found that the expression of DDR genes, involved in repair pathways for DNA double-strand breaks, was upregulated in highly malignant primary and recurrent brain tumors, and their expression was related to abnormal clinical features. However, a potent HDAC6 inhibitor, MPT0B291, attenuated the expression of these genes, including RAD51 and CHEK1, and was more effective in blocking homologous recombination repair in GBM cells. Interestingly, it resulted in lower cytotoxicity in primary glial cells than other HDAC6 inhibitors. MPT0B291 reduced the growth of both TMZ-sensitive and TMZ-resistant tumor cells and prolonged survival in mouse models of GBM. We verified that HDAC6 regulated DDR genes by affecting Sp1 expression, which abolished MPT0B291-induced DNA damage. Our findings uncover a regulatory network among HDAC6, Sp1, and DDR genes for drug resistance and survival of GBM cells. Furthermore, MPT0B291 may serve as a potential lead compound for GBM therapy.


Transconjugation and genotyping of the plasmid-mediated AmpC beta-lactamase and extended-spectrum beta-lactamase genes in Klebsiella pneumoniae.
AmpC beta-lactamases and extended-spectrum beta-lactamases (ESBLs) are becoming predominant causes of resistance to third and forth-generation cephalosporins in Klebsiella pneumoniae (K. pneumoniae). It is very difficult to treat infectious diseases caused by multidrug-resistant K. pneumoniae. The purpose of the present study was to investigate transconjugation and characteristics of beta-lactamase genes in K. pneumoniae producing AmpC beta-lactamases and ESBLs. AmpC beta-lactamases were detected by three-dimension test and ESBLs by disc confirmatory test. Minimum inhibitory concentrations (MICs) were determined by agar dilution. Transfer of resistance to EC600 (Rif(r)) was attempted by conjugation in broth and screened on agar containing cefotaxime (2 microg/ml) plus rifampin (1024 microg/ml). The genes encoding AmpC or ESBLs and their transconjugants were detected by PCR and verified by DNA sequencing. The resistant rates to ampicillin and piperacillin were 100% in 18 isolates of K. pneumoniae. However, imipenem was still of great bactericidal activity on K. pneumoniae, and its MIC(50) was 0.5 microg/mL. Eleven beta-lactamase genes, including TEM-1, TEM-11, SHV-13, SHV-28, CTX-M-9, CTX-M-22, CTX-M-55, OXA-1, LEN, OKP-6 and DHA-1, were found from 18 isolates. And at least one beta-lactamase gene occurred in each isolate. To our surprise, there were six beta-lactamase genes in the CZ04 strain. Among 18 isolates of K. pneumoniae, the partial resistant genes in 8 isolates were conjugated successfully, which had 100% homological sequence with donors by sequence analysis. Compared with donors, 8 transconjugants had attained resistance to most beta-lactams, including ampicillin, piperacillin, cefoxitin, cefotaxime and aztreonam, or even amikacin and gentamicin. R plasmids can be easily transferred between the resistant and sensitive negative bacilli. It is very difficult to block and prevent the spread of antimicrobial resistance. So more attention should be paid to reducing the frequency, times and dosage of antimicrobials, especially third or fourth cephalosporins.


Longitudinal aliasing in multislice helical computed tomography: sampling and cone-beam effects.
In this study, we examine longitudinal aliasing properties in multislice helical computed tomography (CT) volumes reconstructed under the multiple parallel fanbeam approximation by use of a 180LI-type algorithm. We focus on the differences between the multislice case and the single-slice case, which has been studied previously. Specifically, we examine longitudinal aliasing properties in four-slice scanners for helical pitches 3 and 6, which are sometimes called "preferred" in four-slice helical CT, because it is believed that the effective longitudinal sampling intervals at these pitches are equivalent to those in single-slice helical CT operating at pitches 1 and 2, respectively. While these equivalences have been supported by comparative studies of slice-sensitivity profiles in single- and multislice helical CT, artifacts have been observed in pitch-3 and pitch-6 multislice images that were not evident in their purported single-slice counterparts. We attribute these differences to aliasing arising in the multislice reconstructions that is not present in the single-slice counterparts. We find that the aliasing has two principal origins: sampling effects similar to those in the single-slice case and cone-beam effects. The difference between the multislice, pitch-3 and single-slice, pitch-1 results is attributed to the small cone angle in multislice helical CT, which introduces inconsistencies among the measurements of different detector rows. The difference between multislice, pitch-6 and single-slice, pitch-2 results is attributed to a combination of the cone angle and genuine differences in sampling patterns. It is argued, however, that the lack of strict equivalence with single-slice counterparts does not necessarily undermine the claim that pitches 3 and 6 are "preferred" relative to other pitches in multislice helical CT.


Malaria 1984. Part I. Malaria prophylaxis.
Guidelines are presented to assist the prescriber in arriving at a logical and up-to-date approach to the chemoprophylaxis of malaria, particularly for residents of and visitors to Africa. Areas of controversy are highlighted, with a critical but simplified practical approach being taken to a complex subject. The importance of a history of recent travel in a patient presenting with an unexplained fever is stressed. A series of tables listing geographical areas where chloroquine- and other drug-resistant Plasmodium falciparum is found is provided as an aid to the logical selection of prophylactic agents for the traveller. For convenience, tables of dosages for all ages are given. Similarly, an algorithm is provided to facilitate decision-making in drug usage. The controversial topic of malaria prophylaxis in pregnancy is discussed in the light of published information. Although the prophylaxis of possible drug-resistant malaria has been alluded to in Part I, in Part II of this series the treatment and theoretical aspects of resistant malaria will be covered with a view to providing guidelines for those faced with this ever-increasing problem.


Granulocyte colony-stimulating factor administered as a single intraperitoneal injection modifies the lethal dose95/30 in irradiated B6D2F1 mice.
Granulocyte colony-stimulating factor (G-CSF) is a hematopoietic growth factor that stimulates the proliferation of progenitor myeloid cells. We have previously demonstrated that recombinant human G-CSF (rhG-CSF) significantly improves survival of lethally irradiated B6D2F1 mice when administered as a single intraperitoneal dose of 1 mg/kg 2 hours after a lethal dose (LD)95/30 irradiation. In our model, rhG-CSF is also able to modify the LD95/30 in irradiated animals and 1.1 has been found to be the dose modification factor (the ratio of LD95/30 for mice treated with rhG-CSF to that for control animals).


Temporal associations between daytime physical activity and sleep in children.
We examined temporal associations between objectively-measured physical activity (PA) during the day and in the evening, and sleep quantity and quality. PA and sleep were measured by actigraphs for an average of one week in an epidemiological cohort study of 275 eight-year-old children. For each one standard deviation (SD) unit of increased PA during the day, sleep duration was decreased by 0.30, sleep efficiency by 0.16, and sleep fragmentation increased by 0.08 SD units that night. For each one SD unit increase in sleep duration and efficiency the preceding night, PA the following day decreased by 0.09 and 0.16 SD units, respectively. When we contrasted days with a high amount of moderate to vigorous activity during the day or in the evening to days with a more sedentary profile, the results were essentially similar. However, moderate to vigorous PA in the evening shortened sleep latency. The relationship between a higher level of PA and poorer sleep is bidirectional. These within-person findings challenge epidemiological findings showing that more active people report better sleep. Since only a few studies using objective measurements of both PA and sleep have been conducted in children, further studies are needed to confirm/refute these results.


Molecular epidemiology of extended-spectrum β-lactamase-, AmpC β-lactamase- and carbapenemase-producing Escherichia coli and Klebsiella pneumoniae isolated from Canadian hospitals over a 5 year period: CANWARD 2007-11.
To assess the proportion of Escherichia coli and Klebsiella pneumoniae from Canadian hospitals that produce extended-spectrum β-lactamases (ESBLs), AmpC β-lactamases and carbapenemases, as well as to describe the patterns of antibiotic resistance and molecular characteristics of these organisms. Some 5451 E. coli and 1659 K. pneumoniae were collected from 2007 to 2011 inclusive as part of the ongoing CANWARD national surveillance study. Antimicrobial susceptibility testing was performed to detect putative ESBL, AmpC and carbapenemase producers, which were then further characterized by PCR and sequencing to detect resistance genes. In addition, isolates were characterized by PFGE and an allele-specific PCR to detect isolates of sequence type (ST) 131. The proportion of ESBL-producing E. coli (2007, 3.4%; 2011, 7.1%), AmpC-producing E. coli (2007, 0.7%; 2011, 2.9%) and ESBL-producing K. pneumoniae (2007, 1.5%; 2011, 4.0%) among the isolates collected increased during the study period. The majority of ESBL-producing E. coli (>95%), AmpC-producing E. coli (>97%) and ESBL-producing K. pneumoniae (>89%) remained susceptible to colistin, amikacin, ertapenem and meropenem. Isolates were generally unrelated by PFGE (<80% similarity); however, ST131 was identified among 55.8% and 28.7% (P < 0.001) of ESBL- and AmpC-producing E. coli, respectively. CTX-M-15 was the dominant genotype in both ESBL-producing E. coli (66.2%) and ESBL-producing K. pneumoniae (50.0%), while the dominant genotype in AmpC-producing E. coli was CMY-2 (55.7%). Carbapenemase production was identified in 0.04% (n = 2) of E. coli and 0.06% (n = 1) of K. pneumoniae, all of which produced KPC-3. The proportion of ESBL- and AmpC-producing E. coli and K. pneumoniae increased significantly during the study period, while the number of carbapenemase producers remained low (<1%). Compared with AmpC-producing E. coli, ESBL-producing E. coli were significantly associated with multidrug resistance and the ST131 clone.


The use of 3D digital anatomy model improves the communication with patients presenting with prostate disease: The first experience in Senegal.
We hypothesized that the use of an interactive 3D digital anatomy model can improve the quality of communication with patients about prostate disease. A 3D digital anatomy model of the prostate was created from an MRI scan, according to McNeal's zonal anatomy classification. During urological consultation, the physician presented the digital model on a computer and used it to explain the disease and available management options. The experience of patients and physicians was recorded in questionnaires. The main findings were as follows: 308 patients and 47 physicians participated in the study. In the patient group, 96.8% reported an improved level of understanding of prostate disease and 90.6% reported an improved ability to ask questions during consultation. Among the physicians, 91.5% reported improved communication skills and 100% reported an improved ability to obtain patient consent for subsequent treatment. At the same time, 76.6% of physicians noted that using the computer model lengthened the consultation. This exploratory study found that the use of a 3D digital anatomy model in urology consultations was received overwhelmingly favorably by both patients and physicians, and it was perceived to improve the quality of communication between patient and physician. A randomized study is needed to confirm the preliminary findings and further quantify the improvements in the quality of patient-physician communication.


Application of biologically-oriented dentin bonding principles to the use of endodontic irrigants.
To compare the removal of smear layer and the structure of the hybrid layer formed after the use of EDTA or MTAD solutions when used as a final flush. Single-rooted extracted premolars (n=18) were collected and treated for root canal therapy using NaOCl irrigation followed by a final rinse of 17% EDTA, Biopure MTAD, or saline (negative control). The roots were obturated with gutta-percha and a hydrophilic HEMA-containing root canal sealer. The TEM specimens were impregnated with 50% silver nitrate to visualize sealing imperfections and nanoleakage. The structure of the coronal, middle and apical parts of root canal walls was examined using transmission electron microscopy. After NaOCl irrigation, a final rinse with BioPure MTAD or 17% EDTA completely removed the 2 microm-thick smear layer on mechanically instrumented root canal walls. The BioPure MTAD hybrid layer was thicker than the 17% EDTA hybrid layer. Both the BioPure MTAD and EDTA caused a collapse of the dentin matrix structure which impeded sealer infiltration and the formation of high quality hybrid layer bonding. The hybrid layers created in smear layer-covered dentin exhibited less potential for nanoleakage than the MTAD or EDTA hybrid layers.


[Chinese expert consensus on minimally invasive radical surgery for pancreatic ductal adenocarcinoma (version 2022)].
Although there are still controversies over the efficiency and safety of minimally invasive radical surgery for pancreatic ductal adenocarcinoma (PDAC), most available studies have suggested a promising application of minimally invasive radical surgery. This consensus, referring to Chinese expert opinions and worldwide researches, aimed to discuss the related issues on minimally invasive radical surgery for PDAC to ensure the perioperative and oncological outcomes. Quality of evidence and strength of recommendations were evaluated based on the GRADE approach. The 15 recommendations covered 5 topics: oncological outcomes and patient safety of laparoscopic and robotic pancreatoduodenectomy, left-side pancreatectomy for PDAC, learning curve, safety of neoadjuvant therapy, and vascular resection in minimally invasive radical surgery for PDAC. This consensus gives reference and guidance to surgeons on the use of minimally invasive radical surgery for PDAC. Although this consensus is not sufficient to answer all the questions about minimally invasive radical surgery for PDAC, it represents the current consensus on the application of the techniques in the treatment of PDAC on the Chinese mainland.


The amino acid composition of 350 lymphocyte proteins.
We determined the amino acid composition of proteins of Sp2 hybridoma cells by a procedure which assembles the information on the polypeptides upon two-dimensional gel electrophoresis, such that biosynthetic labeling with 20 different 3H amino acids provides the data--spot intensities--on the relative representation of the detected polypeptides. The gels were impregnated with 2,5-diphenyloxazol (PPO) and suitably exposed radiofluorographs were selected for analysis. The images originating from the 12 cultures labeled with amino acids R, A, H, I, L, K, M, F, P, S, T and Y were analysed with the Kepler image analysis system. The spot volume data of the 12 analysed patterns were corrected for the unequal labeling efficiencies of the 3H amino acids and for the various exposure times. This correction is performed by applying calibration factors based on the amino acid determination of a hydrolysate of the analysed cells. After the calibration step was applied to the data files we used the amino acid compositions of nine proteins taken from a database to establish for each of these proteins the correlation coefficients with the image analysis derived amino acid compositions of all spots. The correlation coefficients allow us to tentatively identify polypeptide spots on two-dimensional gels, while the amino acid composition of 350 investigated two-dimensional gel spots is usable as an identification tag in the gene retrieval from our cDNA libraries.


An early Cambrian greenhouse climate.
The oceans of the early Cambrian (~541 to 509 million years ago) were the setting for a marked diversification of animal life. However, sea temperatures-a key component of the early Cambrian marine environment-remain unconstrained, in part because of a substantial time gap in the stable oxygen isotope (δ18O) record before the evolution of euconodonts. We show that previously overlooked sources of fossil biogenic phosphate have the potential to fill this gap. Pristine phosphatic microfossils from the Comley Limestones, UK, yield a robust δ18O signature, suggesting sea surface temperatures of 20° to 25°C at high southern paleolatitudes (~65°S to 70°S) between ~514 and 509 million years ago. These sea temperatures are consistent with the distribution of coeval evaporite and calcrete deposits, peak continental weathering rates, and also our climate model simulations for this interval. Our results support an early Cambrian greenhouse climate comparable to those of the late Mesozoic and early Cenozoic, offering a framework for exploring the interplay between biotic and environmental controls on Cambrian animal diversification.


Focal p53 protein expression and lymphovascular invasion in primary prostate tumors predict metastatic progression.
TP53 is one of the most frequently altered genes in prostate cancer. The precise assessment of its focal alterations in primary tumors by immunohistochemistry (IHC) has significantly enhanced its prognosis. p53 protein expression and lymphovascular invasion (LVI) were evaluated for predicting metastatic progression by IHC staining of representative whole-mounted prostate sections from a cohort of 189 radical prostatectomy patients with up to 20 years of clinical follow-up. Kaplan-Meier survival curves were used to examine time to distant metastasis (DM) as a function of p53 expression and LVI status. TP53 targeted sequencing was performed in ten tumors with the highest expression of p53 staining. Nearly half (49.8%) of prostate tumors examined showed focal p53 expression while 26.6% showed evidence of LVI. p53(+) tumors had higher pathologic T stage, Grade Group, Nuclear Grade, and more frequent LVI. p53 expression of > 5% and LVI, individually and jointly, are associated with poorer DM-free survival. TP53 mutations were detected in seven of ten tumors sequenced. Four tumors with the highest p53 expression harbored likely pathogenic or pathogenic mutations. High levels of p53 expression suggest the likelihood of pathogenic TP53 alterations and, together with LVI status, could enhance early prognostication of prostate cancer progression.


Prevalence and distribution of spotted fever and typhus infections in Sierra Leone and Ivory Coast.
A serosurvey for evidence of rickettsial infections was conducted in the rural populations of several tropical rain forest areas in Sierra Leone and Ivory Coast. Seropositivity rates were surprisingly high in both countries, with more than 7% of the individuals in some districts having antibodies to spotted fever-group rickettsiae. No significant difference was found in the overall prevalence of diagnostic antibody titers to spotted fever-group rickettsiae in Sierra Leone (5.3%) and Ivory Coast (6.2%). However, there was a significant difference (p less than 0.001) in the prevalence of diagnostic antibody titers to typhus rickettsiae in the two countries. There were no marked geographic differences within either country in overall prevalence of rickettsial infections, but there were possible area differences in specific seropositivity rates to typhus- and spotted fever-group rickettsiae in Sierra Leone. In both countries, age and sex differences were important in determining seropositivity, but there was no indication of an age-sex interaction. In Sierra Leone, 59 of the 80 positive sera (73.8%) were from persons age 15 or above (p less than 0.001), and 50 of the 80 (62.5%) were from males (p = 0.05). In Ivory Coast, 33 of the 37 positive sera (89.2%) were from the greater than or equal to 15-age group, and 28 of the 37 (75.7%) were from males (p less than 0.001 for both age and sex). The identification of specific areas endemic for these rickettsial diseases should facilitate the diagnosis and treatment of patients with rickettsial illnesses in West Africa.


After acute coronary syndrome, diabetic patients with peripheral vascular disease remain at high risk of cardiovascular events despite secondary prevention measures.
Peripheral vascular disease (PVD) is associated with a high risk of cardiovascular events after an acute coronary syndrome (ACS). The impact of suboptimal risk-factor control and drug prescription on morbidity and mortality rates in patients with PVD following an ACS remains to be established. To assess whether a global atherosclerosis management programme and optimal secondary prevention could benefit high-risk PVD patients after an ACS. A total of 851 ACS patients underwent an intensified intervention focusing on evaluating risk factors and atherosclerosis lesions, and on optimizing treatment and education. We compared its impact on long-term risk factors, medication observance and cardiovascular outcomes in patients with coronary artery disease (CAD) alone (n=715, 84.0%) and with both CAD and PVD (n=136). At a median follow-up of 18.6months, both groups reached recommended secondary prevention goals and showed no significant differences in rates of drug prescription. PVD was not associated with minor cardiovascular events (hazard ratio [HR] 1.32, 95% confidence interval [CI] 0.57-3.02) but remained independently associated with major (HR 2.15, 95% CI 1.12-4.13) and total (HR 1.76, 95% CI 1.05-2.93) cardiovascular events. Compared to patients with CAD alone, this risk was significantly higher in CAD patients with both PVD and diabetes (HR 2.87, 95% CI 1.52-5.43), but not in PVD patients without diabetes (HR 1.35, 95% CI 0.71-2.56) or diabetic patients without PVD (HR 1.11, 95% CI 0.68-1.81). Despite optimization of risk-factor control and drug prescription after ACS, patients with both PVD and diabetes carry a 2.9-fold higher risk of cardiovascular events at 18-month follow-up versus patients with CAD alone. This excess risk was not significant in PVD patients without diabetes or in diabetic patients without PVD.


A p-Adic model for the process of thinking disturbed by physiological and information noise.
We develop a model of the process of thinking in the presence of noise (which is produced by the simultaneous action of a huge number of neurons in the brain as well as by external information and internal cognitive processes). Our model is based on Freud's idea on the splitting of cognitive processes into two (closely connected) domains: consciousness and subconsciousness. We represent the process of thinking as a random dynamical process in a space of ideas endowed with a non-Euclidean geometry (which differs extremely from the ordinary Euclidean geometry of spatial location of neurons in the brain). The so-called p-adic geometry on a space of ideas describes the ability of cognitive systems to form associations. We show that random dynamical thinking systems on a p -adic space of ideas still generate only deterministic ideas. We also study positive and negative effects of noise (in particular, creativeness and stress).


Electronic and magnetic properties of monolayer α-RuCl3: a first-principles and Monte Carlo study.
Recent experiments revealed that monolayer α-RuCl3 can be obtained by a chemical exfoliation method and exfoliation or restacking of nanosheets can manipulate the magnetic properties of the materials. In this paper, the electronic and magnetic properties of an α-RuCl3 monolayer are investigated by combining first-principles calculations and Monte Carlo simulations. From first-principles calculations, we found that the spin configuration of FM corresponds to the ground state for α-RuCl3, however, the other excited zigzag oriented spin configuration has an energy of 5 meV per atom higher than the ground state. The energy band gap is found to be 3 meV using PBE functionals. When the spin-orbit coupling effect is taken into account, the corresponding energy gap is determined to be 57 meV. We also investigate the effect of the Hubbard U energy terms on the electronic band structure of the α-RuCl3 monolayer and revealed that the band gap increases approximately linearly with increasing U value. Moreover, spin-spin coupling terms (J1, J2, and J3) have been obtained using first-principles calculations. By benefiting from these terms, Monte Carlo simulations with a single site update Metropolis algorithm have been implemented to elucidate the magnetic properties of the considered system. Thermal variations of magnetization, susceptibility and also specific heat curves indicate that monolayer α-RuCl3 exhibits a phase transition between ordered and disordered phases at the Curie temperature of 14.21 K. We believe that this study can be utilized to improve two-dimensional magnetic materials.


Interplay Between Duration of Androgen Deprivation Therapy and External Beam Radiotherapy With or Without a Brachytherapy Boost for Optimal Treatment of High-risk Prostate Cancer: A Patient-Level Data Analysis of 3 Cohorts.
Radiotherapy combined with androgen deprivation therapy (ADT) is a standard of care for high-risk prostate cancer. However, the interplay between radiotherapy dose and the required minimum duration of ADT is uncertain. To determine the specific ADT duration threshold that provides a distant metastasis-free survival (DMFS) benefit in patients with high-risk prostate cancer receiving external beam radiotherapy (EBRT) or EBRT with a brachytherapy boost (EBRT+BT). This was a cohort study of 3 cohorts assembled from a multicenter retrospective study (2000-2013); a post hoc analysis of the Randomized Androgen Deprivation and Radiotherapy 03/04 (RADAR; 2003-2007) randomized clinical trial (RCT); and a cross-trial comparison of the RADAR vs the Deprivación Androgénica y Radio Terapía (Androgen Deprivation and Radiation Therapy; DART) 01/05 RCT (2005-2010). In all, the study analyzed 1827 patients treated with EBRT and 1108 patients treated with EBRT+BT from the retrospective cohort; 181 treated with EBRT and 203 with EBRT+BT from RADAR; and 91 patients treated with EBRT from DART. The study was conducted from October 15, 2020, to July 1, 2021, and the data analyses, from January 5 to June 15, 2021. High-dose EBRT or EBRT+BT for an ADT duration determined by patient-physician choice (retrospective) or by randomization (RCTs). The primary outcome was DMFS; secondary outcome was overall survival (OS). Natural cubic spline analysis identified minimum thresholds (months). This cohort study of 3 studies totaling 3410 men (mean age [SD], 68 [62-74] years; race and ethnicity not collected) with high-risk prostate cancer found a significant interaction between the treatment type (EBRT vs EBRT+BT) and ADT duration (binned to <6, 6 to <18, and ≥18 months). Natural cubic spline analysis identified minimum duration thresholds of 26.3 months (95% CI, 25.4-36.0 months) for EBRT and 12 months (95% CI, 4.9-36.0 months) for EBRT+BT for optimal effect on DMFS. In RADAR, the prolongation of ADT for patients receiving only EBRT was not associated with significant improvements in DMFS (hazard ratio [HR], 1.01; 95% CI, 0.65-1.57); however, for patients receiving EBRT+BT, a longer duration was associated with improved DMFS (DMFS HR, 0.56; 95% CI, 0.36-0.87; P = .01). For patients receiving EBRT alone (DART), 28 months of ADT was associated with improved DMFS compared with 18 months (RADAR HR, 0.37; 95% CI, 0.17-0.80; P = .01). These cohort study findings suggest that the optimal minimum ADT duration for treatment with high-dose EBRT alone is more than 18 months; and for EBRT+BT, it is 18 months or possibly less. Additional studies are needed to determine more precise minimum durations.


The somatostatinergic system in the mammalian cochlea.
Little is known about expression and function of the somatostatinergic system in the mammalian cochlea. We have previously shown that somatostatin administration may have a protective effect on gentamicin-induced hair cell loss. In this study, we have analyzed the cochlear expression of somatostatin receptor 1 (SST1) and somatostatin receptor 2 (SST2) at both the mRNA and the protein level in wild-type mice, as well as in SST1 and SST2 knock-out (KO) mice and in cultivated neurosensory cells. We demonstrate that the somatostatin receptors SST1 and SST2 are specifically expressed in outer and inner hair cells (HCs) of the organ of Corti (OC), as well as in defined supporting cells. The expression of SST1 and SST2 receptors in cultivated P5 mouse OC explants was similar to their expression in inner and outer hair cells. Somatostatin itself was not expressed in the mammalian cochlea, suggesting that somatostatin reaches its receptors either through the blood-labyrinthine barrier from the systemic circulation or via the endolymphatic duct from the endolymphatic sac. We used mice with a deletion of either SST1 or SST2 to learn more about the regulation of SST1 and SST2 receptor expression. We demonstrate that in SST1 KO mice, SST2 was expressed in outer HCs and Deiters' cells, but not in pillar cells or inner HCs, as compared with wild-type mice. In contrast, in SST2 KO mice, the expression pattern of the SST1 receptor was not altered relative to wild-type mice. These findings reveal that somatostatin receptors demonstrate specific expression in HCs and supporting cells of the mouse cochlea, and that absence of SST1 alters the expression of SST2. This specific expression pattern suggests that somatostatin receptors may have important functional roles in the inner ear.


[Solid-phase immunoenzyme ass for detecting Giardia lamblia antigens].
An ELISA sandwich for the detection of Giardia lamblia antigens in human faeces was standardized. 175 samples were studied: 77 positive, 61 negative to cysts and/or trophozoites by direct faeces test, and 19 positive to other parasite different from G. lamblia. The sensitivity of the technique was 94.8% and the specificity 98.3%. The method detects an antigen concentration of 31 ng. The procedure is simple, sensitive and specific so, it may be useful for diagnosis and in epidemiological studies.


Overnight orthokeratology lens wear can inhibit the central stromal edema response.
To investigate the overnight corneal edema response during overnight orthokeratology (OK). Eighteen young adult myopic subjects wore reverse-geometry lenses in Boston XO material (nominal Dk/t 46 x 10(-9) cm.mL O2/s.mL.mmHg) on an overnight wearing schedule for 1 month. Another 10 subjects wore conventional rigid gas-permeable (GP) lenses of similar Dk/t in one eye only, on an identical schedule. Corneal stromal thicknesses in the center, midperiphery, and periphery were measured by optical pachometry in the morning after lens removal, after 1, 4, 10, and 30 nights of wear. Changes from baseline for OK, GP and no-lens eyes were compared by repeated-measures ANOVA and protected post hoc t-tests. The central stroma swelled significantly less in OK than in GP eyes (P < 0.001, ANOVA), and less than with no lens wear (P < 0.001, ANOVA) throughout the study. Overnight edema levels consistent with Dk/t were found on day 1 in the midperiphery (3.5 mm from apex) and periphery (5.0 mm) with both OK and GP lenses. The overnight edema response decreased significantly through the study with both lens types. Recovery to baseline stromal thickness during the day was demonstrated for GP eyes and for OK eyes in the central and peripheral cornea. Overnight wear of reverse-geometry OK lenses inhibited the central stromal edema response. Overnight edema levels consistent with Dk/t were found in the corneal midperiphery and periphery. Adaptation of the edema response occurred with continuing overnight lens wear. The results suggest that central pressure exerted by the flat-fitting base curve of the OK lens acts locally as a "clamp" to inhibit overnight central corneal swelling.


Deep denitrification: Stream and groundwater biogeochemistry reveal contrasted but connected worlds above and below.
Excess nutrients from agricultural and urban development have created a cascade of ecological crises around the globe. Nutrient pollution has triggered eutrophication in most freshwater and coastal ecosystems, contributing to a loss in biodiversity, harm to human health, and trillions in economic damage every year. Much of the research conducted on nutrient transport and retention has focused on surface environments, which are both easy to access and biologically active. However, surface characteristics of watersheds, such as land use and network configuration, often do not explain the variation in nutrient retention observed in rivers, lakes, and estuaries. Recent research suggests subsurface processes and characteristics may be more important than previously thought in determining watershed-level nutrient fluxes and removal. In a small watershed in western France, we used a multi-tracer approach to compare surface and subsurface nitrate dynamics at commensurate spatiotemporal scales. We combined 3-D hydrological modeling with a rich biogeochemical dataset from 20 wells and 15 stream locations. Water chemistry in the surface and subsurface showed high temporal variability, but groundwater was substantially more spatially variable, attributable to long transport times (10-60 years) and patchy distribution of the iron and sulfur electron donors fueling autotrophic denitrification. Isotopes of nitrate and sulfate revealed fundamentally different processes dominating the surface (heterotrophic denitrification and sulfate reduction) and subsurface (autotrophic denitrification and sulfate production). Agricultural land use was associated with elevated nitrate in surface water, but subsurface nitrate concentration was decoupled from land use. Dissolved silica and sulfate are affordable tracers of residence time and nitrogen removal that are relatively stable in surface and subsurface environments. Together, these findings reveal distinct but adjacent and connected biogeochemical worlds in the surface and subsurface. Characterizing how these worlds are linked and decoupled is critical to meeting water quality targets and addressing water issues in the Anthropocene.


Complications associated with isotretinoin use after rhinoplasty.
This study aimed to evaluate the association between the postoperative use of isotretinoin and the formation of nasal tip deformities after rhinoplasty. A retrospective study investigated a case series of patients who presented with nasal tip deformities subsequent to the use of isotretinoin after rhinoplasty. Patients who had taken isotretinoin after rhinoplasty were identified from a single surgeon's case log. Clinic charts and operative reports were reviewed. Predisposing factors and time intervals to complications were identified. Three patients were found to have undergone isotretinoin therapy after rhinoplasty. Nasal tip deformities included bossa formation, asymmetry, and prominence of a composite graft. All three patients required subsequent procedures to repair soft tissue nasal tip deformities. In all three cases, isotretinoin was started within 2 years of the primary rhinoplasty. The nasal tip deformities were first observed within 6 months after isotretinoin was started. Three cases are described in which postoperative use of isotretinoin was associated with nasal tip deformities after rhinoplasty. Further studies are needed to investigate this possible causative relationship and to describe the risks involved.


Forty Years of Dengue Surveillance at a Tertiary Pediatric Hospital in Bangkok, Thailand, 1973-2012.
Long-term observational studies can provide valuable insights into overall dengue epidemiology. Here, we present analysis of dengue cases at a pediatric hospital in Bangkok, Thailand, during a 40-year period from 1973 to 2012. Data were analyzed from 25,715 hospitalized patients with laboratory-confirmed dengue virus (DENV) infection. Several long-term trends in dengue disease were identified including an increase in mean age of hospitalized cases from an average of 7-8 years, an increase after 1990 in the proportion of post-primary cases for DENV-1 and DENV-3, and a decrease in the proportion of dengue hemorrhagic fever and dengue shock syndrome cases in primary and post-primary cases over time. Exploratory mechanistic analysis of these observed trends considered changes in diagnostic methods, demography, force of infection, and Japanese encephalitis vaccination as possible explanations. Thailand is an important setting for studying DENV transmission as it has a "mature" dengue epidemiology with a strong surveillance system in place since the early 1970s. We characterized changes in dengue epidemiology over four decades, and possible impact of demographic and other changes in the human population. These results may inform other countries where similar changes in transmission and population demographics may now or may soon be occurring.


Challenges of Implementing an Individual Randomized Controlled Trial (Women First: Preconception Maternal Nutrition Study) in a Rural Study Site: A Case Study From Pakistan.
Persistent global disparities in maternal and neonatal outcomes associated with poor maternal nutrition provided the genesis of the Women First (WF) study, an individually randomized controlled trial on preconceptional maternal nutrition. This article describes the challenges that arose in implementing this trial related to nutrition or diet of the mother, in District Thatta-Pakistan. During different phases of the study, we encountered problems in identifying the eligible participants, taking consent from couples, randomizing participants in different arms, conducting biweekly follow-up visits on time, ensuring compliance to the intervention, and measuring the primary outcome within the 24 hours of birth. Each challenge was itself an opportunity for the research team to address the same through effective coordination and teamwork. Moreover, with adequate resources and dedicated staff with diverse backgrounds, it was possible to implement the WF study across the widely scattered geographic clusters of District Thatta. In addition, there are some broad strategies that could be applied to other studies such as very close contact either in person or at least by talking to mothers via phones and rapport with the study participants, the study leadership of country coordinator and the field supervisors to build trust between those on front lines and the study leadership. Moreover, continuous monitoring and supervision with frequent training and refreshers were also found to be more important to assure the data quality and to meet the study targets. Community meetings were also found to be very helpful and effective to follow the participants for a long time. Researchers conducting a similar type of studies particularly in rural areas can learn many lessons from such experiences. Thus, the process of implementing the study in one of the rural areas of Pakistan provides an insight into where and how similar individual randomized trials might be deployed.


Social support and risk of compulsory admission: part IV of the Amsterdam Study of Acute Psychiatry.
Social support for patients with a mental illness has been associated with a lower rate of hospitalization. It is important to clarify the role played by a lack of social support as a possible predictor of emergency compulsory admission. A random sample of 252 patients who were evaluated by two psychiatric emergency teams in Amsterdam from September 2004 to September 2006 were interviewed approximately one month later about their social networks and social interactions. The number of emergency compulsory admissions was recorded for 244 patients during a two-year follow-up period after the interviews. Patients who lived alone had a higher risk of compulsory admission (p≤.05) and had fewer people in their social network (4.6 versus 6.1, p≤.001) compared with patients who lived with others. Among patients who lived alone, the percentage of patients with a compulsory admission was significantly higher among the patients with a high score for negative interactions than among patients with a low score (34% versus 13%, p≤.05). Of the social support variables, living alone proved to be the only predictor of emergency compulsory admission and readmission, and patients who lived alone had a smaller social network. A high level of negative social interactions increased the risk of compulsory admission among patients who lived alone.


Electron microscopic characterization of chick embryonic skeletal muscle proteoglycans.
In this article, proteoglycans from embryonic chick leg muscle are quantitatively and qualitatively compared with day 8 high density cell culture cartilage proteoglycans by electron microscopy of proteoglycan-cytochrome c monolayers. The visualized proteoglycan profiles were separated into four categories according to shape, size, and complexity. The two major categories were further characterized by lengths of core proteins, lengths of side projections, and distance between side projections. Two large proteoglycans are identifiable in spread leg muscle preparations. One group has a core protein (mean length of 205 nm) from which extend long thin side projections that we interpret to be groups of chondroitin sulfate glycosaminoglycans with a mean length of 79 nm. This large chondroitin sulfate proteoglycan is the only type found in muscle cultures as determined both biochemically in the past and now by electron microscopy and is referred to as muscle proteoglycan. The second large proteoglycan has a mean core protein length of 250 nm and side projections that are visibly shorter (mean length of 38 nm) and thicker than those of the muscle proteoglycan. This group is referred to as the mesenchymal proteoglycan since its biosynthetic origin is still uncertain. We compare these two profiles with the chick cartilage chondroitin sulfate proteoglycan that has a mean core protein length of 202 nm and side projections with a mean length of 50 nm. The data presented here substantiate the earlier biochemical characterization of these noncartilage proteoglycans and establish the unique structural features of the muscle proteoglycan as compared with the similar profiles of the cartilage and mesenchymal proteoglycans.


Optical density of the human lens.
Optic disk reflectance was measured from 27 normal observers with their physiological lenses (aged 21-74 yr) and from two pseudophakic observers (aged 69 and 70 yr) with use of a Utrecht fundus reflection densitometer. Psychophysical heterochromatic flicker photometric luminance matches (10 degrees field) were obtained on the same group of the observers. A four-parameter model incorporating lens density, hemoglobin absorption, optic disk reflectance, and superficial stray light was used to fit the reflectometric data. A model incorporating lens density and the Judd revised spectral luminous-efficiency function was used to fit the psychophysical data. The lens-density spectrum used the two-factor aging model of Pokorny, et al. [Appl. Opt. 26, 1437 (1987)]. The lens density for each normal observer was estimated through a least-squares fitting procedure yielding an estimated lens age. For the reflectometric data the observer's chronological age agreed with estimated lens age with a correlation coefficient of 0.92. The reflectometric regression line underestimated chronological age by approximately 5 yr. The mean reflectance of the optic disk was 0.047 with standard error of the mean of 0.0044. Data from the pseudophakic observers were well described when corneal density was used to replace lens density. The lens density was also estimated from the psychophysical data. The observer's chronological age agreed with psychophysically estimated lens age with a correlation coefficient of 0.92. It was concluded that the in vivo lens density can be estimated from the reflectance spectrum measured off the optic disk. The reflectance spectrum of the optic disk was inferred to be close to spectrally neutral.


SdhA blocks disruption of the Legionella-containing vacuole by hijacking the OCRL phosphatase.
Legionella pneumophila grows intracellularly within a replication vacuole via action of Icm/Dot-secreted proteins. One such protein, SdhA, maintains the integrity of the vacuolar membrane, thereby preventing cytoplasmic degradation of bacteria. We show here that SdhA binds and blocks the action of OCRL (OculoCerebroRenal syndrome of Lowe), an inositol 5-phosphatase pivotal for controlling endosomal dynamics. OCRL depletion results in enhanced vacuole integrity and intracellular growth of a sdhA mutant, consistent with OCRL participating in vacuole disruption. Overexpressed SdhA alters OCRL function, enlarging endosomes, driving endosomal accumulation of phosphatidylinositol-4,5-bisphosphate (PI(4,5)P2), and interfering with endosomal trafficking. SdhA interrupts Rab guanosine triphosphatase (GTPase)-OCRL interactions by binding to the OCRL ASPM-SPD2-Hydin (ASH) domain, without directly altering OCRL 5-phosphatase activity. The Legionella vacuole encompassing the sdhA mutant accumulates OCRL and endosomal antigen EEA1 (Early Endosome Antigen 1), consistent with SdhA blocking accumulation of OCRL-containing endosomal vesicles. Therefore, SdhA hijacking of OCRL is associated with blocking trafficking events that disrupt the pathogen vacuole.


Managing the security of nursing data in the electronic health record.
The Electronic Health Record (EHR) is a patient care information resource for clinicians and nursing documentation is an essential part of comprehensive patient care. Ensuring privacy and the security of health information is a key component to building the trust required to realize the potential benefits of electronic health information exchange. This study was aimed to manage nursing data security in the EHR and also discover the viewpoints of hospital information system vendors (computer companies) and hospital information technology specialists about nursing data security. This research is a cross sectional analytic-descriptive study. The study populations were IT experts at the academic hospitals and computer companies of Tehran city in Iran. Data was collected by a self-developed questionnaire whose validity and reliability were confirmed using the experts' opinions and Cronbach's alpha coefficient respectively. Data was analyzed through Spss Version 18 and by descriptive and analytic statistics. The findings of the study revealed that user name and password were the most important methods to authenticate the nurses, with mean percent of 95% and 80%, respectively, and also the most significant level of information security protection were assigned to administrative and logical controls. There was no significant difference between opinions of both groups studied about the levels of information security protection and security requirements (p>0.05). Moreover the access to servers by authorized people, periodic security update, and the application of authentication and authorization were defined as the most basic security requirements from the viewpoint of more than 88 percent of recently-mentioned participants. Computer companies as system designers and hospitals information technology specialists as systems users and stakeholders present many important views about security requirements for EHR systems and nursing electronic documentation systems. Prioritizing of these requirements helps policy makers to decide what to do when planning for EHR implementation. Therefore, to make appropriate security decisions and to achieve the expected level of protection of the electronic nursing information, it is suggested to consider the priorities of both groups of experts about security principles and also discuss the issues seem to be different between two groups of participants in the research.


Androgen receptor expression in prostate cancer cells is suppressed by activation of epidermal growth factor receptor and ErbB2.
Prostate cancers (PCa) that relapse after androgen deprivation therapies [castration-resistant PCa (CRPC)] express high levels of androgen receptor (AR) and androgen-regulated genes, and evidence from several groups indicates that ErbB family receptor tyrosine kinases [epidermal growth factor (EGF) receptor (EGFR) and ErbB2] may contribute to enhancing this AR activity. We found that activation of these kinases with EGF and heregulin-beta1 rapidly (within 8 hours) decreased expression of endogenous AR and androgen-regulated PSA in LNCaP PCa cells. AR expression was similarly decreased in LAPC4 and C4-2 cells, but not in the CWR22Rv1 PCa cell line. The rapid decrease in AR was not due to increased AR protein degradation and was not blocked by phosphatidylinositol 3-kinase (LY294002) or MEK (UO126) inhibitors. Significantly, AR mRNA levels in LNCaP cells were markedly decreased by EGF and heregulin-beta1, and experiments with actinomycin D to block new mRNA synthesis showed that AR mRNA degradation was increased. AR mRNA levels were still markedly decreased by EGF and heregulin-beta1 in LNCaP cells adapted to growth in androgen-depleted medium, although AR protein levels did not decline due to increased AR protein stability. These findings show that EGFR and ErbB2 can negatively regulate AR mRNA and may provide an approach to suppress AR expression in CRPC.


[Pulmonary artery blood pressure and snoring].
Snoring is an extrathoracic inspiratory airway obstruction during sleep, accompanied by more or less intense noise. The inspiration volume being limited at the same time, this can produce additional blood gas changes. Epidemiological studies have shown an increased risk of myocardial infarction or cerebral ischaemia during night for snorers. Little is known, however, about acute effects of snoring on the haemodynamics of heart and circulation. The present study presents data of the pulmonary artery blood pressure during sleep-related upper airway obstruction. Pulmonary arterial pressure during sleep was examined with five patients via Swan-Ganz catheter. When an upper airway obstruction occurred, the inspiratory pulmonary arterial pressures rose in all the five patients. Two patients showed a clinically significant decrease in arterial oxygen saturation during snoring so that in these cases, a hypoxic vasoconstriction can be discussed as a possible cause. The other three patients did not produce significant decreases in oxygen saturation, so that in these cases mechanical factors (negative inspiratory intrathoracic pressure oscillations) should be regarded as the most probable cause for increases in the pulmonary arterial blood pressure.


A Survey of Prescribers' Attitudes, Knowledge, Comfort, and Fear of Consequences Related to an Opioid Overdose Education and Naloxone Distribution Program.
The alarming increase in opioid-related fatalities has placed the United States in the midst of an opioid epidemic. In an effort to reduce opioid-related fatalities, each Veterans Affairs (VA) hospital is responsible for development of an OEND (Overdose Education and Naloxone Distribution) program to enhance naloxone distribution. To assess attitudes, knowledge, comfort level, and fear of consequences at the prescriber level related to the OEND program at VA Connecticut Healthcare System (VACHS). Anonymous surveys were administered to VACHS prescribers to assess attitude, knowledge, comfort level, and fear of consequences using a 5-point Likert scale. VACHS. VACHS prescribers (primary care physicians/internists, psychiatrists, physician assistants/advanced practice registered nurses, pharmacists) in primary care/medicine, mental health, and substance use disorder (SUD). Differences across 4 domains were analyzed using analysis of variance testing, and pairwise post hoc analysis was conducted using Tukey's HSD for all significant findings. Regression modeling was conducted to evaluate correlations between each group and survey responses. Identify differences and potential barriers to program implementation across professions and practice specialty. While primary care/medicine, mental health, and SUD prescribers had positive attitudes toward dispensing naloxone, SUD clinicians reported being more comfortable, more knowledgeable, and less fearful of consequences. In general, psychiatrists reported to be more knowledgeable and comfortable prescribing naloxone. In addition, prescribers who received naloxone training reported themselves to be more comfortable, more knowledgeable, and less fearful of the consequences of dispensing naloxone. Results indicate that barriers to OEND implementation continue to exist, and not all clinicians or practice specialties at VACHS are comfortable with dispensing naloxone. A targeted approach to training health care clinicians may lead to increased acceptance of naloxone dispensing. Additional research is needed to elicit the best method of improved acceptance of naloxone to expand naloxone access.


Sequence analysis of genome segment 10 encoding the major outer capsid protein (VP7) of genogroup B aquareovirus and its relationship with the VP7 protein of genogroup A aquareovirus.
Genome segment 10 of the coho salmon (CSR) strain of genogroup B aquareovirus has been cloned and sequenced. The gene is at least 936 nucleotides in length and has a major open reading frame encoding a protein of 293 amino acids with a calculated molecular mass of 31.7 kDa. Comparison of nucleotide and deduced amino acid sequences of genome segment 10 of genogroup A and genogroup B strains indicated 55.7 and 36.5% identity at the nucleotide and amino acid level, respectively. Baculovirus expression and serological identification of the genome segment 10 product of the CSR strain indicated that this gene encodes the major outer capsid protein of genogroup B aquareovirus.


Reconsidering Cluster Bias in Multilevel Data: A Monte Carlo Comparison of Free and Constrained Baseline Approaches.
The test for item level cluster bias examines the improvement in model fit that results from freeing an item's between level residual variance from a baseline model with equal within and between level factor loadings and between level residual variances fixed at zero. A potential problem is that this approach may include a misspecified unrestricted model if any non-invariance is present, but the log-likelihood difference test requires that the unrestricted model is correctly specified. A free baseline approach where the unrestricted model includes only the restrictions needed for model identification should lead to better decision accuracy, but no studies have examined this yet. We ran a Monte Carlo study to investigate this issue. When the referent item is unbiased, compared to the free baseline approach, the constrained baseline approach led to similar true positive (power) rates but much higher false positive (Type I error) rates. The free baseline approach should be preferred when the referent indicator is unbiased. When the referent assumption is violated, the false positive rate was unacceptably high for both free and constrained baseline approaches, and the true positive rate was poor regardless of whether the free or constrained baseline approach was used. Neither the free or constrained baseline approach can be recommended when the referent indicator is biased. We recommend paying close attention to ensuring the referent indicator is unbiased in tests of cluster bias. All Mplus input and output files, R, and short Python scripts used to execute this simulation study are uploaded to an open access repository.


Recurrent pulmonary infections in patients with chronic lymphocytic leukemia and hypogammaglobulinemia.
We describe two patients with concomitant recurrent pulmonary infections due to encapsulated organisms and chronic lymphocytic leukemia and hypogammaglobulinemia. We discuss the cases as a subset of acquired hypogammaglobulinemia in adults, focusing on the possible role of impaired defense mechanisms and emphasizing the importance of aggressive therapy with gamma globulin, antibiotics, and good pulmonary toilet to reduce morbidity.