Genomic characterization of two Chinese isolates of porcine respiratory and reproductive syndrome virus.
The genomes of two isolates of porcine respiratory and reproductive syndrome virus (PRRSV) from China, designated HB-1(sh)/2002 and HB-2(sh)/2002, were sequenced and analyzed. The size of the genomes of HB-1(sh)/2002 and HB-2(sh)/2002 were 15,411 and 15,373 nucleotides respectively, excluding the poly(A) tails. Comparative analysis with the genomic sequences of another Chinese isolate (BJ-4) and North American (VR2332) and European (Lelystad virus, LV) viruses revealed that HB-1(sh)/2002 shared 89.8% identity with BJ-4 and VR2332, but only 54.7% with LV; while HB-2(sh)/2002 shared 89.4% and 89.5% identity with BJ-4 and VR2332 respectively and 54.3% with LV, indicating that the two new Chinese isolates were related to the North American PRRSV genotype. Phylogenetic analysis based on the nucleotide sequence of the structural protein ORF's showed that the two new Chinese isolates belong to same genetic subgroup. HB-2(sh)/2002 additionally exhibited variations in the NSP2 nonstructural protein encoded by ORF1 and the structural protein GP3 encoded by ORF3 in comparison with other North American PRRSV isolates, namely a 12 amino acids deletion in Nsp2 and one amino acid deletion in GP3 were found in HB-2(sh)/2002. Therefore, HB-2(sh)/2002 was a novel strain with unique deletions.


Novel ionically crosslinked acrylamide-grafted poly(vinyl alcohol)/sodium alginate/sodium carboxymethyl cellulose pH-sensitive microspheres for delivery of Alzheimer's drug donepezil hydrochloride: Preparation and optimization of release conditions.
In this work, the graft copolymer, poly(vinyl alcohol)-grafted polyacrylamide (PVA-g-PAAm), was synthesized and characterized by Fourier transform infrared spectroscopy, differential scanning calorimetry, and elemental analysis. Microspheres of PVA-g-PAAm/sodium alginate (NaAlg)/sodium carboxymethyl cellulose (NaCMC) were prepared by the emulsion-crosslinking method and used for the delivery of an Alzheimer's drug, donepezil hydrochloride (DP). The release of DP increased with the increase in drug/polymer ratio (d/p) and PVA-g-PAAm/NaAlg/NaCMC ratio, while it decreased with the increase in the extent of crosslinking. The optimum DP release was obtained as 92.9% for a PVA-g-PAAm/NaAlg/NaCMC ratio of 1/2/1, d/p ratio of 1/8, and FeCl3 concentration of 7% (w/v).


Pancreatic carcinoma masked as fever of unknown origin: A case report and comprehensive review of literature.
Pancreatic carcinoma is a highly lethal malignancy. Common presenting features of pancreatic cancer include anorexia, asthenia, weight loss, pain, and obstructive jaundice. Nevertheless, fever as a symptom, or even primary manifestation of pancreatic cancer is rather rare. Here, we report a 63-year-old male patient presenting with daily fevers, night sweats, and fatigue of 2-month duration. Laboratory findings showed elevated white blood cell count (WBC), erythrocyte sedimentation rate (ESR), and serum C-reactive protein (CRP). A computed tomography scan demonstrated a tumor between the duodenum and pancreatic head. Chest radiograph was normal. The patient underwent an uneventful tumor resection. Histological examination of a surgical specimen demonstrated an undifferentiated adenocarcinoma originated from pancreatic head. The tumor was compatible with TNM stage IIA (T3N0M0). Complete resolution of the fever was achieved on post-operative day 4 and no recurrence of the tumor or neoplastic fever happen during the 39-month follow-up. Pancreatic adenocarcinoma could manifest as neoplastic fever at the time of diagnosis. If the tumor is resectable, surgical resection is a safe and curative form of therapy not only for the fever but also for the original carcinoma.


Structure-Based Discovery of Potent, Orally Bioavailable Benzoxazepinone-Based WD Repeat Domain 5 Inhibitors.
The chromatin-associated protein WDR5 (WD repeat domain 5) is an essential cofactor for MYC and a conserved regulator of ribosome protein gene transcription. It is also a high-profile target for anti-cancer drug discovery, with proposed utility against both solid and hematological malignancies. We have previously discovered potent dihydroisoquinolinone-based WDR5 WIN-site inhibitors with demonstrated efficacy and safety in animal models. In this study, we sought to optimize the bicyclic core to discover a novel series of WDR5 WIN-site inhibitors with improved potency and physicochemical properties. We identified the 3,4-dihydrobenzo[f][1,4]oxazepin-5(2H)-one core as an alternative scaffold for potent WDR5 inhibitors. Additionally, we used X-ray structural analysis to design partially saturated bicyclic P7 units. These benzoxazepinone-based inhibitors exhibited increased cellular potency and selectivity and favorable physicochemical properties compared to our best-in-class dihydroisoquinolinone-based counterparts. This study opens avenues to discover more advanced WDR5 WIN-site inhibitors and supports their development as novel anti-cancer therapeutics.


Distinct effects of saracatinib on memory CD8+ T cell differentiation.
Immunologic memory involving CD8(+) T cells is a hallmark of an adaptive Ag-specific immune response and constitutes a critical component of protective immunity. Designing approaches that enhance long-term T cell memory would, for the most part, fortify vaccines and enhance host protection against infectious diseases and, perhaps, cancer immunotherapy. A better understanding of the cellular programs involved in the Ag-specific T cell response has led to new approaches that target the magnitude and quality of the memory T cell response. In this article, we show that T cells from TCR transgenic mice for the nucleoprotein of influenza virus NP68 exhibit the distinct phases--priming, expansion, contraction, and memory--of an Ag-specific T cell response when exposed in vitro to the cognate peptide. Saracatinib, a specific inhibitor of Src family kinases, administered at low doses during the expansion or contraction phases, increased CD62L(high)/CD44(high) central memory CD8(+) T cells and IFN-γ production but suppressed immunity when added during the priming phase. These effects by saracatinib were not accompanied by the expected decline of Src family kinases but were accompanied by Akt-mammalian target of rapamycin suppression and/or mediated via another pathway. Increased central memory cells by saracatinib were recapitulated in mice using a poxvirus-based influenza vaccine, thus underscoring the importance of dose and timing of the inhibitor in the context of memory T cell differentiation. Finally, vaccine plus saracatinib treatment showed better protection against tumor challenge. The immune-potentiating effects on CD8(+) T cells by a low dose of saracatinib might afford better protection from pathogens or cancer when combined with vaccine.


Influence of erroneous patient records on population pharmacokinetic modeling and individual bayesian estimation.
Observational data sets can be used for population pharmacokinetic (PK) modeling. However, these data sets are generally less precisely recorded than experimental data sets. This article aims to investigate the influence of erroneous records on population PK modeling and individual maximum a posteriori Bayesian (MAPB) estimation. A total of 1123 patient records of neonates who were administered vancomycin were used for population PK modeling by iterative 2-stage Bayesian (ITSB) analysis. Cut-off values for weighted residuals were tested for exclusion of records from the analysis. A simulation study was performed to assess the influence of erroneous records on population modeling and individual MAPB estimation. Also the cut-off values for weighted residuals were tested in the simulation study. Errors in registration have limited the influence on outcomes of population PK modeling but can have detrimental effects on individual MAPB estimation. A population PK model created from a data set with many registration errors has little influence on subsequent MAPB estimates for precisely recorded data. A weighted residual value of 2 for concentration measurements has good discriminative power for identification of erroneous records. ITSB analysis and its individual estimates are hardly affected by most registration errors. Large registration errors can be detected by weighted residuals of concentration.


Using a Rasch Model to Account for Guessing as a Source of Low Discrimination.
The most common approach to modelling item discrimination and guessing for multiple-choice questions is the three parameter logistic (3PL) model. However, proponents of Rasch models generally avoid using the 3PL model because to model guessing entails sacrificing the distinctive property and advantages of Rasch models. One approach to dealing with guessing based on the application of Rasch models is to omit responses in which guessing appears to play a significant role. However, this approach entails loss of information and it does not account for variable item discrimination. It has been shown, though, that provided specific constraints are met, it is possible to parameterize discrimination while preserving the distinctive property of Rasch models. This article proposes an approach that uses Rasch models to account for guessing on standard multiple-choice items simply by treating it as a source of low item discrimination. Technical considerations are noted although a detailed examination of such considerations is beyond the scope of this article.


Carbon dioxide and trace oxygen concentrations impact growth and product formation of the gut bacterium Phocaeicola vulgatus.
The promising yet barely investigated anaerobic species Phocaeicola vulgatus (formerly Bacteroides vulgatus) plays a vital role for human gut health and effectively produces organic acids. Among them is succinate, a building block for high-value-added chemicals. Cultivating anaerobic bacteria is challenging, and a detailed understanding of P. vulgatus growth and metabolism is required to improve succinate production. One significant aspect is the influence of different gas concentrations. CO2 is required for the growth of P. vulgatus. However, it is a greenhouse gas that should not be wasted. Another highly interesting aspect is the sensitivity of P. vulgatus towards O2. In this work, the effects of varying concentrations of both gases were studied in the in-house developed Respiratory Activity MOnitoring System (RAMOS), which provides online monitoring of CO2, O2, and pressure under gassed conditions. The RAMOS was combined with a gas mixing system to test CO2 and O2 concentrations in a range of 0.25-15.0 vol% and 0.0-2.5 vol%, respectively. Changing the CO2 concentration in the gas supply revealed a CO2 optimum of 3.0 vol% for total organic acid production and 15.0 vol% for succinate production. It was demonstrated that the organic acid composition changed depending on the CO2 concentration. Furthermore, unrestricted growth of P. vulgatus up to an O2 concentration of 0.7 vol% in the gas supply was proven. The viability decreased rapidly at concentrations larger than or equal to 1.3 vol% O2. The study showed that P. vulgatus requires little CO2, has a distinct O2 tolerance and is therefore well suited for industrial applications.


[Age sensitivity characteristics of the human and mouse chromosome apparatus in exposure to viral infection].
It is determined that the measles virus (vaccine strain Leningrad-16) induce cytogenetic alterations in bone marrow cells in all the examined mice, the most considerable aberrations being noted in newly born and especially aged mice. Coxsackie virus A13 is capable of causing cytogenetic aberrations in newly born and aged mice, while the puberal mice of the middle aged group are resistant to the mutagenic effect of the virus. Flu patients aged 65-80 and 2-5 years suffer from the biggest alterations in the blood chromosome leucocytes, with the decrease in number and in T-lymphocyte functional activity. The patients aged 18-35 years suffered less.


Sonoelectrosynthesis of monodisperse metal nanoparticles.
Traditional colloidal syntheses of metal nanoparticles (NPs) are highly sensitive to the selection of and quality of chemical reducing agents and metal precursors. To address these challenges, we demonstrate the complete sonoelectrochemical synthesis of monodisperse metal NPs starting from bulk metal, using Cu as a model system. Electrochemical syntheses of NPs are of great interest as the oxidation and reduction processes that account for product formation can occur directly at the anode and cathode, respectively. This ability has the potential to improve reproducibility by simplifying the chemical pathway to NPs, with electrosyntheses often also providing unique kinetic pathways toward green product formation. Herein, ultrasound is coupled with electrosynthesis to clean the electrode surface, dispersing the NPs produced at the electrode into solution. We were able to shift the size distribution to form monodispersed metal NPs through control of applied potential (Vapplied) and ultrasonic pulses. The synthesis begins with electrooxidation of bulk Cu metal to directly dissolve metal ions into a microemulsion system. This step is followed by sonoelectroreduction of the ions, which facilitates the formation of dispersible, monodisperse Cu NPs with diameters <10 nm. The size distribution can be controlled by adjusting the Vapplied, pulse intensity, and pulse sequence implemented during sonoelectroreduction. We view this technique as a scalable method to synthesize metal NPs from bulk metal without chemical reducing agents.


Cool storage of Queensland fruit fly eggs for increased flexibility in rearing programs.
The Queensland fruit fly (Q-fly) is Australia's most economically damaging insect pest of fruit crops. The Sterile Insect Technique (SIT) used to suppress outbreaks relies on supply of high-quality flies and this can be assisted by the ability to manipulate production schedules. Cool storage at temperatures that are sufficient to slow development without causing significant somatic damage can provide a valuable means of manipulating production schedules. In this study, we investigate the effect of four storage temperatures (10, 13, 16 and 19 °C) and three exposure times (3, 6 or 9 days) on Q-fly eggs. Egg storage proved effective in prolonging the developmental time of Q-flies. Storage at 10 °C was unsuitable, resulting in a low hatching rate for all exposure times. Hatching rate was also significantly reduced when eggs were exposed to 13 °C for 6 or 9 days, followed by a significant reduction in the number of pupae recovered. Storage at 16 °C yielded promising results, prolonging the preimaginal development of Q-flies up to 6.5 days without significantly affecting egg hatching or subsequent development. Cool storage of eggs shows promise as a tool for prolonging the development of Q-flies to manipulate schedules in mass rearing programs. © 2018 Society of Chemical Industry.


CUSUM charts for monitoring clinical practice quality using primary care prescribing data: a case study of an initiative to encourage generic prescribing of proton pump inhibitors.
Expectations on organizations to monitor quality of care are growing. Whilst relevant data are increasingly becoming available it is, in many cases, difficult to distinguish real effects from background variation. Here, the potential usefulness of cumulative sum (CUSUM) charts for monitoring the use of medicines is explored through a case study of an initiative to encourage the prescribing of lowest cost proton pump inhibitors (PPI) in the context of implementation of national guidelines for the management of dyspepsia. This was a longitudinal study involving analysis of routinely collected prescribing data, set in all 12 primary care trusts (PCT) in the North East Strategic Health Authority. In it, comparison (by subtraction) of the time-series of the percentage of generic PPI prescription items for Gateshead with the mean of the other 11 PCTs was used to reduce both variation and bias. This was followed by the construction of a CUSUM chart displaying the effect of the Gateshead initiative. The simple process of comparison was very successful both in removing extraneous trends and reducing background variation, and the CUSUM highly effective for displaying the evidence for the hypothesized step-change in prescribing behaviour consequent on the Gateshead initiative. The effectiveness of the CUSUM here is strongly linked to the success of the preliminary comparison step. CUSUM, a statistical process control technique, has already been tested as a tool for interpreting hospital and general practice mortality rates. Here, its potential for more general applications in quality monitoring is demonstrated using routinely collected prescribing data. Such data contain valuable information about changes in clinical practice and CUSUM charts, when coupled with the idea of removing time trends and extraneous variation by reference to average behaviour, can provide a simple but effective technique for extracting it.


Disruption and overexpression of Arabidopsis phytosulfokine receptor gene affects cellular longevity and potential for growth.
Phytosulfokine (PSK), a 5-amino acid sulfated peptide that has been identified in conditioned medium of plant cell cultures, promotes cellular growth in vitro via binding to the membrane-localized PSK receptor. Here, we report that loss-of-function and gain-of-function mutations of the Arabidopsis (Arabidopsis thaliana) PSK receptor gene (AtPSKR1) alter cellular longevity and potential for growth without interfering with basic morphogenesis of plants. Although mutant pskr1-1 plants exhibit morphologically normal growth until 3 weeks after germination, individual pskr1-1 cells gradually lose their potential to form calluses as tissues mature. Shortly after a pskr1-1 callus forms, it loses potential for growth, resulting in formation of a smaller callus than the wild type. Leaves of pskr1-1 plants exhibit premature senescence after bolting. Leaves of AtPSKR1ox plants exhibit greater longevity and significantly greater potential for callus formation than leaves of wild-type plants, irrespective of their age. Calluses derived from AtPSKR1ox plants maintain their potential for growth longer than wild-type calluses. Combined with our finding that PSK precursor genes are more strongly expressed in mature plant parts than in immature plant parts, the available evidence indicates that PSK signaling affects cellular longevity and potential for growth and thereby exerts a pleiotropic effect on cultured tissue in response to environmental hormonal conditions.


Association of Repetitive Transcranial Magnetic Stimulation Treatment With Subgenual Cingulate Hyperactivity in Patients With Major Depressive Disorder: A Secondary Analysis of a Randomized Clinical Trial.
Hyperactivity in the subgenual cingulate cortex (SGC) is associated with major depressive disorder (MDD) and anticorrelated with activity in the dorsolateral prefrontal cortex (DLPFC). This association was found to be predictive of responsiveness to repetitive transcranial magnetic stimulation (rTMS) treatment. Such findings suggest that DLPFC-SGC connectivity is important for understanding both the therapeutic mechanism of rTMS in patients with MDD and the underlying pathophysiology of MDD. To evaluate SGC hyperactivity in patients with MDD before and after rTMS treatment. In this diagnostic study, among participants recruited from the adult and geriatric mood and anxiety services at the Centre for Addiction and Mental Health, Toronto, Ontario, Canada, who had participated in a randomized clinical trial, baseline SGC activity of patients with MDD was compared with healthy controls. In patients with MDD, SGC activity was compared before and after active or sham high-frequency rTMS treatment. Data collection started in July 2008 and concluded in March 2012. Neurophysiological data analysis started in January 2017 and ended in May 2018. Hyperactivity in the SGC before and after rTMS treatment was measured. Subgenual cingulate cortex hyperactivity activity was quantified using significant current density (SCD), and effective connectivity between the left DLPFC and SGC was computed using significant current scattering (SCS). Both measures were computed around TMS evoked potentials standard peak latencies prior to rTMS and after rTMS treatment, comparing patients with MMD treated with active and sham rTMS. Patients with MDD were assessed with the 17-item Hamilton Rating Scale for Depression. Of 121 patients with MDD in the initial trial, 30 (15 [50.0%] women) were compared with 30 healthy controls (15 [50.0%] women) at rTMS treatment baseline. The mean (SD) age of the cohort with MDD was 39.1 (10.9) years, and the mean (SD) age of healthy controls was 37.0 (11.0) years. Following rTMS treatment, 26 patients with MDD who had active rTMS treatment (21.5%) were compared with 17 patients with MDD who had sham treatment (14.0%). At baseline, the SGC mean (SD) SCD and mean (SD) SCS at 200 milliseconds after TMS pulse were higher in participants with MDD compared with healthy controls (SCD: 1.04 × 10-6 [1.41 × 10-6] μA/mm2 vs 3.8 × 10-7 [7.8 × 10-7] μA/mm2; z = -2.95; P = .004; SCS: 0.87 [0.86] mm vs 0.54 [0.87] mm; z = -2.27; P = .02). Baseline source current density was able to classify MDD with 77% accuracy. Scores on the 17-item Hamilton Rating Scale for Depression were correlated with current density at the SGC (ρ = 0.41; P = .03). After rTMS treatment, SGC mean (SD) SCD and mean (SD) SCS at 200 milliseconds after rTMS pulse were attenuated to approximately the standard TMS-evoked potential latencies in the active rTMS group compared with the sham rTMS group (SCD: 1.57 × 10-7 [3.67 × 10-7] μA/mm2 vs 7.00 × 10-7 [7.51 × 10-7] μA/mm2; z = -2.91; P = .004; SCS: 0.20 [0.44] mm vs 0.74 [0.73] mm; z = -2.78; P = .006). Additionally, the SGC SCS change was correlated with symptom improvement on the 17-item Hamilton Rating Scale for Depression in the active rTMS group (ρ = 0.58; P = .047). The findings of this study further implicate left DLPFC-SGC effective connectivity and SGC excitability in the pathophysiology of MDD and treatment with rTMS. These findings suggest that DLPFC-SGC connectivity may be a marker of rTMS treatment responsiveness. ClinicalTrials.gov identifier: NCT01515215.


Electronic recording of occlusion treatment for amblyopia: potential of the new technology.
Approximately one third of all amblyopic eyes do not reach visual acuity of 20/40 in spite of occlusion therapy. One of the reasons is a lack of adherence to therapy, which, however, could not be quantified in the past. Experience with new devices (occlusion dose monitors, ODMs) for electronic recording of occlusion has recently been reported. The aim of the present study was to evaluate the potential of the ODMs developed in The Netherlands. Various features were tested, including the reliability of the ODM recordings compared to diaries, two ODMs used simultaneously on one patch, the influence of the ambient temperature, and the specificity of the recording pattern for measurements on the eye. The ODMs were taped to the outside of the standard occlusion patch and measured the temperature difference between their front and back surfaces. Members of the research group and the families of two patients kept occlusion diaries while using the ODMs. Recorded and written occlusion periods were compared. Measurements were carried out under various conditions: patch with one ODM tightly on the eye or detached (allowing peeping); ODMs taped to various parts of the body; two ODMs simultaneously on one patch; variation of room temperature. There was good correspondence between the occlusion times recorded by the ODMs and those from the diaries, as well as between the recordings of two ODMs used simultaneously on one patch. High ambient temperatures (33 degrees C to 37 degrees C) prevented reliable ODM measurements. Measurements on other parts of the body were misclassified with probabilities between P=0.099 and P=0.325 as measurements with the patch tightly on the eye. In spite of some technical limitations, the ODMs provide a chance for reliable assessment of compliance and therefore objective information on dose-response function for occlusion therapy. This will lead to a more evidence-based treatment for amblyopia.


Lethal 17D yellow fever encephalitis in mice. I. Passive protection by monoclonal antibodies to the envelope proteins of 17D yellow fever and dengue 2 viruses.
Monoclonal antibodies to the envelope proteins (E) of the 17D vaccine strain of yellow fever virus (17D YF) and to dengue 2 virus were examined for their ability to confer passive protection against lethal 17D YF encephalitis in mice. All 13 IgG anti-17D YF antibodies, regardless of neutralizing capacity, conferred solid protection when given in a relatively high dose prior to intracerebral inoculation of virus. Three antibodies with high in vitro neutralizing titres were all protective at a low dose as were several non-neutralizing antibodies. One flavivirus group-reactive antibody to dengue 2 virus conferred similar protection at low dose. Protection was also observed when antibodies were given several days after virus inoculation when peak infectious virus titres and histopathological evidence of infection were present in brains. The ability of a non-neutralizing antibody to protect could not be attributed to complement-dependent lysis of virus-infected cells and did not correlate with avidity or with proximity of its binding site to a critical neutralizing epitope of the E protein. Some antibodies, characterized as non-neutralizing by plaque reduction assay on Vero cells, inhibited the growth of virus in a mouse neuroblastoma cell line, suggesting one possible mechanism of protection. These results may be relevant to the design of prospective flavivirus vaccines and support the possibility of conferring broadened protection among flaviviruses by stimulating the antibody response to appropriate epitopes of the E protein.


Deriving a probabilistic syntacto-semantic grammar for biomedicine based on domain-specific terminologies.
Biomedical natural language processing (BioNLP) is a useful technique that unlocks valuable information stored in textual data for practice and/or research. Syntactic parsing is a critical component of BioNLP applications that rely on correctly determining the sentence and phrase structure of free text. In addition to dealing with the vast amount of domain-specific terms, a robust biomedical parser needs to model the semantic grammar to obtain viable syntactic structures. With either a rule-based or corpus-based approach, the grammar engineering process requires substantial time and knowledge from experts, and does not always yield a semantically transferable grammar. To reduce the human effort and to promote semantic transferability, we propose an automated method for deriving a probabilistic grammar based on a training corpus consisting of concept strings and semantic classes from the Unified Medical Language System (UMLS), a comprehensive terminology resource widely used by the community. The grammar is designed to specify noun phrases only due to the nominal nature of the majority of biomedical terminological concepts. Evaluated on manually parsed clinical notes, the derived grammar achieved a recall of 0.644, precision of 0.737, and average cross-bracketing of 0.61, which demonstrated better performance than a control grammar with the semantic information removed. Error analysis revealed shortcomings that could be addressed to improve performance. The results indicated the feasibility of an approach which automatically incorporates terminology semantics in the building of an operational grammar. Although the current performance of the unsupervised solution does not adequately replace manual engineering, we believe once the performance issues are addressed, it could serve as an aide in a semi-supervised solution.


Expression of simple epithelial type cytokeratins in stratified epithelia as detected by immunolocalization and hybridization in situ.
Multi-layered ("stratified") epithelia differ from one-layered ("simple") polar epithelia by various architectural and functional properties as well as by their cytoskeletal complements, notably a set of cytokeratins characteristic of stratified tissue. The simple epithelial cytokeratins 8 and 18 have so far not been detected in any stratified epithelium. Using specific monoclonal antibodies we have noted, in several but not all samples of stratified epithelia, including esophagus, tongue, exocervix, and vagina, positive immunocytochemical reactions for cytokeratins 8, 18, and 19 which in some regions were selective for the basal cell layer(s) but extended into suprabasal layers in others. In situ hybridization with different probes (riboprobes, synthetic oligonucleotides) for mRNAs of cytokeratin 8 on esophageal epithelium has shown, in extended regions, relatively strong reactivity for cytokeratin 8 mRNA in the basal cell layer. In contrast, probes to cytokeratin 18 have shown much weaker hybridization which, however, was rather evenly spread over basal and suprabasal strata. These results, which emphasize the importance of in situ hybridization in studies of gene expression in complex tissues, show that the genes encoding simple epithelial cytokeratins can be expressed in stratified epithelia. This suggests that continual expression of genes coding for simple epithelial cytokeratins is compatible with the formation of squamous stratified tissues and can occur, at least in basal cell layers, simultaneously with the synthesis of certain stratification-related cytokeratins. We also emphasize differences of expression and immunoreactivity of these cytokeratins between different samples and in different regions of the same stratified epithelium and discuss the results in relation to changes of cytokeratin expression during fetal development of stratified epithelia, in response to environmental factors and during the formation of squamous cell carcinomas.


Predictors of non-adherence to aromatase inhibitors among commercially insured women with breast cancer.
The purpose of this study is to investigate 1-year adherence rates to aromatase inhibitors (AI) and to determine risk factors for non-adherence among commercially insured post-menopausal breast cancer patients. A retrospective cohort of 13,593 commercially insured breast cancer patients with a prescription claim for an AI therapy (exemestane, anastrozole, and letrozole) in 2006 were identified using the MarketScan Commercial Claims and Encounters Database. Adherence was calculated by the medication possession ratio (MPR) for a 1-year period following the initial claim in 2006. The main outcome variable was non-adherence (<80% MPR) to AI therapy. Multivariate logistic regression was used to determine predictors of non-adherence. Over a 1-year period, 23% of patients were non-adherent with their AI therapy. AI non-adherence was associated with younger age, out-of-pocket costs ≥$30 per AI prescription, as well as with measures during the 12-month period prior to the initial 2006 AI claim of lower patient out-of-pocket total pharmacy costs, no mastectomy, and higher Charlson Comorbidity Index. Non-adherence to AI is a common occurrence. A number of factors were identified that influence patience non-adherence. These factors can be used to identifying patients at increased risk for non-adherence to better target and intervene to support medication taking behaviors.


The clinical profile of children in India with pigmentary anomalies along the lines of Blaschko and central nervous system manifestations.
The aim of this study was to describe the clinical profile of children with congenital pigmentary anomalies along Blaschko lines and the associated manifestations in the central nervous system. Twenty-six children aged 12 years or less (14 boys and 12 girls), with hypopigmentation and hyperpigmentation along Blaschko lines and central nervous system manifestations were included during the period June 2001 to December 2003. Detailed physical, cutaneous, and systemic examinations were carried out. Relevant investigations included histopathology, karyotyping, electroencephalogram, computerized tomography scans, and magnetic resonance imaging of the brain whenever possible. Twenty children had hyperpigmentation along the lines of Blaschko, four had hypopigmentation, and two had a combination of the two. Eight children had diffuse involvement. Of these, two each had hypomelanosis of Ito, incontinentia pigmenti and linear and whorled nevoid hypermelanosis. The remaining 20 patients could not be categorized as any of the described entities. The majority (92.3%) of patients manifested skin and central nervous system disorders before the age of 2 years. Hyperpigmentation along Blaschko lines was significantly higher in patients with central nervous system manifestations (p = 0.01). Developmental delay was the most frequent central nervous system presentation. Multiple systems were affected, including the eyes. Histology was useful to distinguish incontinentia pigmenti from other types of nevoid hyperpigmentation.


Preclinical In Vitro Assessment of Submicron-Scale Laser Surface Texturing on Ti6Al4V.
Loosening of orthodontic and orthopedic implants is a critical and common clinical problem. To minimize the numbers of revision surgeries due to peri-implant inflammation or insufficient osseointegration, developments of new implant manufacturing strategies are indicated. Ultrafast laser surface texturing is a promising contact-free technology to modify the physicochemical properties of surfaces toward an anti-infectious functionalization. This work aims to texture Ti6Al4V surfaces with ultraviolet (UV) and green (GR) radiation for the manufacturing of laser-induced periodic surface structures (LIPSS). The assessment of these surface modifications addresses key aspects of topography, morphology and chemical composition. Human primary mesenchymal stromal cells (hMSCs) were cultured on laser-textured and polished Ti6Al4V to characterize the surfaces in terms of their in vitro biocompatibility, cytotoxicity, and metal release. The outcomes of the in vitro experiment show the successful culture of hMSCs on textured Ti6Al4V surfaces developed within this work. Cells cultured on LIPSS surfaces were not compromised in terms of their viability if compared to polished surfaces. Yet, the hMSC culture on UV-LIPSS show significantly lower lactate dehydrogenase and titanium release into the supernatant compared to polished. Thus, the presented surface modification can be a promising approach for future applications in orthodontics and orthopedics.


The IgM and IgG response to Bordetella pertussis vaccination and infection.
Ultracentrifugation was used to separate IgG from IgM in serum samples from children after pertussis vaccination or infection and the fractions were examined by indirect haemagglutination (IHA) and complement-fixation (CF) tests. IHA detected pertussis-specific IgM and IgG but CF detected only IgG which appeared to be of a different subclass from the IHA IgG. The IgM titres were higher after infection than after vaccination but the reverse tended to apply to the IgG titres. Little IgM or CF IgG was detected 5 months after a dose of vaccine, but the IHA IgG persisted longer. Vaccinated children who were subsequently infected showed IgM and CF IgG responses similar to those of unvaccinated, infected children but the IHA IgG titres reached much higher levels.


Changes in pediatric dental coverage and visits following the implementation of the affordable care act.
To determine the impact of the inclusion of pediatric dental care in the Affordable Care Act's (ACA) core package of essential health benefits on dental coverage and utilization. Children aged 1-18 years included in the nationally representative 2010-2015 National Health Interview Survey (NHIS). We used regression-adjusted difference-in-differences to examine changes in rates of dental coverage and visits pre- and post-ACA for children likely to be affected by the ACA (have a parent working for a small employer) to a comparison group of children who were unlikely to be affected (have a parent in a firm with ≥50 employees). Models adjusted for relevant health and sociodemographic measures. NHIS is an annual household survey conducted by the National Center for Health Statistics. Comparing pre- and post-ACA periods, private dental insurance increased by 4.6 percentage points more (P = 0.013) and annual dental visits were unchanged (2.7 percentage points, P = 0.071) among children likely to be affected by the ACA compared to children unlikely to be affected by the ACA. Inclusion of pediatric dental care as an ACA essential health benefit increased dental insurance coverage, but not dental visits among children likely to be affected by this policy.


Development of an objective method for the comparison of fired projectiles using an air pistol as a template.
The ability to objectify ballistic evidence is a challenge faced by firearms examiners around the world. A number of researchers are trying to improve bullet-identification systems to address deficiencies detailed within the National Academy of Science report (2009). More recently focus has turned to making use of more sophisticated imaging modalities to view entire regions of the projectile and the development of automated systems for the comparison of the topographical surfaces recorded. Projectiles from a newly bought air pistol with 0.177 calibre pellets (unjacketed), fired series of 609 pellets were examined using an optical microscope. A mathematical methodology was developed to pre-process the resultant topographical maps generating point data for comparison, analysed using the principal component analysis (PCA). In most cases limited to reasonable success was achieved. The objective method still requires an operator to identify the Land Engraved Areas to be scanned, however the mathematical alignments were objectively achieved. The PCA results illustrated that the striation marks were neither exclusive nor specific to the LEA regions but rather crossed over regions. This study also proves that a single weapon does not necessarily leave identical marks of projectiles on its surface.


Risk of developing antiphospholipid antibodies following viral infection: a systematic review and meta-analysis.
Objective The objective of this paper is to conduct a systematic review and meta-analysis on the risk of developing elevated antiphospholipid (aPL) antibodies and related thromboembolic and/or pregnancy events following a viral infection. Method We searched Medline, EMBASE, Web of Science, PubMed ePubs, and Cochrane Central Register of Controlled Trials through June 2016. Independent observational studies of elevated aPL antibodies in patients with a viral infection compared with controls or patients with lupus were included. Results We analyzed 73 publications for 60 studies. Human immunodeficiency virus (HIV) and hepatitis C virus (HCV) were most commonly reported. Compared with healthy controls, patients with HIV were more likely to develop elevated anticardiolipin (aCL) antibodies (risk ratio (RR) 10.5, 95% confidence interval (CI) 5.6-19.4), as were those with HCV (RR 6.3, 95% CI 3.9-10.1), hepatitis B virus (HBV) (RR 4.2, 95% CI 1.8-9.5), and Epstein-Barr virus (EBV) (RR 10.9 95% CI 5.4-22.2). The only statistically significant increased risk for anti-β2-glycoprotein I (anti-β2-GPI) antibodies was observed in patients with HCV (RR 4.8 95% CI 1.0-22.3). Compared with patients with lupus, patients with HIV were more likely to develop elevated aCL antibodies (RR 1.8, 95% CI 1.3-2.6), and those with EBV, elevated anti-β2-GPI antibodies (RR 2.2, 95% CI 1.3-3.9). Thromboembolic events were most prevalent in patients with elevated aPL antibodies who had HCV (9.1%, 95% CI 3.0-18.1), and HBV (5.9%, 95% CI 2.0-11.9) infections, and pregnancy events were most prevalent in those with parvovirus B19 (16.3%, 95% CI 0.78-45.7). However, compared to virus-infected patients with negative aPL antibodies, the only statistically significant increased risk was observed in those with HCV and positive aPL. Conclusions Viral infection can increase the risk of developing elevated aPL antibodies and associated thromboembolic events. Results are contingent on the reported information.


Genetic, embryonic and anatomical study of an interspecific cassava hybrid.
A molecular, anatomical and cytogenetic study of an interspecific hybrid between Manihot esculenta (cassava) and the wild species M. oligantha was carried out. Cytogenetics revealed relatively complete chromosome pairing and high viability of the pollen grains. Ovule structure examined by the clearing method showed polyembryony in 2.7% of the ovules. Doubling of the chromosome number resulted in an increase in polyembryony of up to 18% and a reduction in pollen viability. Multivalent formation was also observed. An anatomical study of stems of diploid and tetraploid hybrids showed a larger number of vascular bundles in the tetraploid type.


Plasminogen activator inhibitor-1 and apolipoprotein E gene polymorphisms and diabetic angiopathy.
A point mutation in the plasminogen activator inhibitor-1 (PAI-1) gene and a three-allelic variation in the apolipoprotein-E (ApoE) gene have been suggested as risk factors for the development of diabetic micro- and macrovascular complications. We studied 198 type 1 diabetic patients with diabetic nephropathy [121 men, age (mean+/-SD) 41+/-10 years, diabetes duration 28+/-8 years] and 192 patients with persistent normoalbuminuria (118 men, age 43+/-10 years, diabetes duration 27+/-9 years). Male patients with nephropathy had elevated plasma PAI-1 levels [geometric mean (95% CI)], 70 (62-79) ng/ml, compared with normoalbuminuric men, 43 (38-47) ng/ml, P<0.001. Even though nephropathic patients with the 4G4G genotype tended to have higher plasma PAI-1 levels, P=0.06, no difference in allele frequency (4G/5G) was seen between patients with and without nephropathy: 0.538/0.462 vs 0.539/0.461, respectively. Nor did ApoE allele frequencies (epsilon2/epsilon3/epsilon4) differ between nephropathic and normoalbuminuric patients: 0.099/0.749/0. 152 vs 0.081/0.745/0.174, respectively. Genotype distributions were also similar, n.s. Coronary heart disease was more prevalent (36%) among nephropathic patients carrying the atherogenic epsilon4-allele compared with 12% in patients with the epsilon3,epsilon3 genotype, P<0.001. No associations between diabetic retinopathy and PAI-1 or ApoE polymorphisms were observed, n.s. The ApoE polymorphism may accelerate the development of coronary heart disease often seen in Caucasian patients with type 1 diabetes and diabetic nephropathy, a condition characterized by elevated plasma PAI-1 in men. Neither the PAI-1 nor the ApoE gene polymorphism contributes to the genetic susceptibility to diabetic nephropathy or retinopathy.


The role of health care providers in long term and complicated workers' compensation claims.
Health care providers (HCPs) play a central role in workers' compensation systems. In most systems, they are involved in the legitimization of work-related injury, are required to provide information to workers' compensation boards about the nature and extent of the injury, give recommendations about return-to-work capability and provide treatment for injury or illness. This study identifies problems that occur at the interface between the health care system, injured workers, and workers' compensation boards (WCBs) that may complicate and extend workers' compensation claims and the mechanisms that underlie the development of these problems. Interviews were sought with injured workers, peer helpers and service providers from a variety of geographic locations in order to get a broad picture of return to work problems and concerns. This analysis includes data from total of 34 interviews with injured workers who had long term and complicated claims. Interviews were also conducted with 14 peer helpers and 21 service providers. We identified four domains related to injured workers' interface with the health care system that played a key role in complicating and prolonging compensation claims. These problems, related to health care access, conflicting or imperfect medical knowledge, limited understanding of compensation system requirements and confusion about decision-making authority, resulted in frustration, financial difficulties and mental health problems for injured workers. Recommendations are made about how compensation system parties can find better ways to serve injured worker health care needs and facilitate a smooth relationship between the compensation board and HCPs.


Septic arthritis. Staphylococcal induction of chondrocyte proteolytic activity.
We report herein that cartilage proteolytic activity increased in bovine and rabbit articular cartilage after treatment with a purified staphylococcal culture medium or intraarticular infection with Staphylococcus aureus. Staphylococcal culture medium increased the release of gelatinolytic, collagenolytic, and caseinolytic activity into the medium of isolated chondrocytes or cartilage organ culture. The proteolytic activities were determined in assays using radiolabeled substrate and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Staphylococcal culture medium was proteolytically inactive by both assay techniques. RNA synthesis of isolated chondrocytes was unaffected by staphylococcal culture medium, whereas overall protein synthesis was inhibited by 84%. An analysis of extracts of Staphylococcus aureus-infected rabbit knee cartilage by substrate gels showed increased gelatinolytic and caseinolytic activity compared with extracts of uninfected knee cartilage. Our data suggest that the rapid loss of proteoglycan and persistent degradation of cartilage in staphylococcal septic arthritis is due to the production and activation of chondrocyte proteases.


Examining the relationship between nutrition and cerebral structural integrity in older adults without dementia.
The proportion of adults aged 60 years and over is expected to increase over the coming decades. This ageing of the population represents an important health issue, given that marked reductions to cerebral macro- and microstructural integrity are apparent with increasing age. Reduced cerebral structural integrity in older adults appears to predict poorer cognitive performance, even in the absence of clinical disorders such as dementia. As such, it is becoming increasingly important to identify those factors predicting cerebral structural integrity, especially factors that are modifiable. One such factor is nutritional intake. While the literature is limited, data from available cross-sectional studies indicate that increased intake of nutrients such as B vitamins (for example, B6, B12 and folate), choline, n-3 fatty acids and vitamin D, or increased adherence to prudent whole diets (for example, the Mediterranean diet) predicts greater cerebral structural integrity in older adults. There is even greater scarcity of randomised clinical trials investigating the effects of nutritional supplementation on cerebral structure, though it appears that supplementation with B vitamins (B6, B12 and folic acid) or n-3 fatty acids (DHA or EPA) may be beneficial. The current review presents an overview of available research examining the relationship between key nutrients or adherence to select diets and cerebral structural integrity in dementia-free older adults.