A multicenter nationwide reference intervals study for common biochemical analytes in Turkey using Abbott analyzers.
A nationwide multicenter study was organized to establish reference intervals (RIs) in the Turkish population for 25 commonly tested biochemical analytes and to explore sources of variation in reference values, including regionality. Blood samples were collected nationwide in 28 laboratories from the seven regions (≥400 samples/region, 3066 in all). The sera were collectively analyzed in Uludag University in Bursa using Abbott reagents and analyzer. Reference materials were used for standardization of test results. After secondary exclusion using the latent abnormal values exclusion method, RIs were derived by a parametric method employing the modified Box-Cox formula and compared with the RIs by the non-parametric method. Three-level nested ANOVA was used to evaluate variations among sexes, ages and regions. Associations between test results and age, body mass index (BMI) and region were determined by multiple regression analysis (MRA). By ANOVA, differences of reference values among seven regions were significant in none of the 25 analytes. Significant sex-related and age-related differences were observed for 10 and seven analytes, respectively. MRA revealed BMI-related changes in results for uric acid, glucose, triglycerides, high-density lipoprotein (HDL)-cholesterol, alanine aminotransferase, and γ-glutamyltransferase. Their RIs were thus derived by applying stricter criteria excluding individuals with BMI >28 kg/m2. Ranges of RIs by non-parametric method were wider than those by parametric method especially for those analytes affected by BMI. With the lack of regional differences and the well-standardized status of test results, the RIs derived from this nationwide study can be used for the entire Turkish population.


Thymectomy in Pemphigus Foliaceus: A Thirty-Year Observation.
Myasthenia gravis is an archetypal human autoimmune disease. Thymectomy is proven effective by controlled clinical trials, and is commonly part of the immunotherapeutic approach when myasthenia creates generalized weakness. Pemphigus foliaceus is also autoimmune but treated medically; and thymectomy is not part of therapy unless thymoma is discovered. Autoimmune mechanisms, age distribution, and response to therapy in autoimmune disorders are likely to be different with thymoma. The concurrence of generalized myasthenia with disfiguring pemphigus foliaceus in one young patient but without thymoma offered a natural experiment to assess immunologic antibody responsiveness postoperatively, and observe more than a quarter of a century of clinical remission of both following total thymectomy surgery.


Impact of initial plasma presepsin level for clinical outcome in hospitalized patients with pneumonia.
Presepsin, the soluble CD14 subtype, is known as a sepsis biomarker. However, its clinical significance in pneumonia is unclear. We investigated the effects of plasma presepsin level on clinical outcomes in patients with pneumonia. Patients over 18 years old admitted to our hospital due to pneumonia from May 2016 through November 2017 were reviewed using electronic medical records. One hundred and seventy-two patients who underwent measurement of plasma presepsin levels on admission were enrolled. Median age of enrolled patients was 81 years [interquartile range (IQR), 68-86 years]. Pneumonia severity index (PSI) class and A-DROP score on admission were calculated. The receiver operating characteristic (ROC) curve analysis was performed to assess the prognostic value of 30-day mortality and to identify the optimal cut-off value of plasma presepsin level. Correlations between plasma presepsin level and other factors were assessed using the Spearman's test. The Kaplan-Meier survival analysis and the log-rank test were performed to assess the two curves differentiated with the optimal cut-off value of plasma presepsin level. Seventeen patients (9.9%) died within 30 days of admission. The deceased patients had higher value of plasma presepsin on admission (539 pg/mL; IQR, 414-832 pg/mL) compared with the survivors (334 pg/mL; IQR, 223-484 pg/mL) (P=0.001). The areas under ROC curve for predicting 30-day mortality were 0.742 for plasma presepsin, 0.755 for A-DROP score, and 0.774 for PSI class. Plasma presepsin level was not associated with etiology of pneumonia. However, it was moderately correlated with serum creatinine level (rs =0.524, P<0.001). The ROC curve analysis derived 470 pg/mL of plasma presepsin level as the optimal cut-off value for predicting 30-day mortality. The Kaplan-Meier survival analysis showed that patients with plasma presepsin level ≥470 pg/mL on admission had significantly higher 30-day mortality than those with plasma presepsin level <470 pg/mL (P<0.001). Among patients with A-DROP score ≥3, those with plasma presepsin level ≥470 mg on admission had significantly higher 30-day mortality (P=0.013). Similarly, among patients with PSI class ≥4, those with plasma presepsin level ≥470 mg on admission had significantly higher 30-day mortality (P=0.005). In hospitalized pneumonia patients, plasma presepsin level on admission could be a useful predictor of 30-day mortality and an additional prognostic biomarker on existing severity assessment scales.


Insertional Torque in Cervical Vertebrae Lateral Mass Screw Fixation: Magerl Technique Versus Roy-Camille Technique.
This is a prospective multicenter comparison study. To measure the insertional torque (IT) for cervical vertebra lateral mass screw (LMS) fixation using the Magerl and Roy-Camille methods, at the discretion of the surgeon. Current fixation methods for patients with preoperative cervical spine instability use LMSs; however, few studies have closely examined intraoperative IT. Patients' bone quality was not measured. A total of 637 posterior cervical multiaxial screws were inserted for LMS (C3-C6) (Magerl technique, 423; Roy-Camille technique, 214) in 107 cases. Patients' mean age was 66 years. Patients treated with the Magerl method were divided into 2 groups, with the men in the MM group and the women in the MF group. Similarly, patients treated with the Roy-Camille method were divided into 2 groups, with the men in the RM group and the women in the RF group. The contralateral cortex was penetrated, and the IT at cerclage was measured at the last time. IT of the lateral mass screw was 53.8±22.4, 45.4±21, 45.5±16.9, and 34±16.4 cN.m in the MM group (258), MF group (165), RM group (163), and RF group (51), respectively. The MM group had a significantly higher IT than the other 3 groups, and the RM group had a significantly higher IT than the RF group. When the correlation between screw length and IT was evaluated, IT was significantly (P<0.05) higher with a longer screw by the Magerl method. The IT was higher with the Magerl method with a longer screw. Screw length and IT were not correlated with the Roy-Camille method; furthermore, the Roy-Camille method went through bilateral cortical bone perpendicularly, so that IT was determined by the fixation power in the cortical part of the bone, which was not thought to be affected by screw length. Level III.


Transarterial therapies for the treatment of intrahepatic cholangiocarcinoma.
Cholangiocarcinoma, whether arising from the intrahepatic or extrahepatic biliary system, is a rare but devastating malignancy. Prognosis is poor, with 5-year overall survival <5% including patients undergoing surgery. Resection is the only curative treatment; however, only ∼30% of patients present at a resectable stage, and intrahepatic recurrence is common even after complete resection. This article discusses the current role of transarterial therapies in the treatment of intrahepatic cholangiocarcinoma.


Occult cocaine and opiate exposure in children and associated physical findings.
We determined the prevalence of cocaine and opiate exposure and the association of exposure with objective physical findings in children presenting to an urban pediatric emergency department. The study included 942 children between one and 60 months of age who required urinalysis for investigation of their chief complaint. Anonymously and without informed consent, urine was screened for benzoylecgonine (BE) and opiates, using an enzyme multiplied immunoassay technique (EMIT) with sensitivity of 50 ng/ml. EMIT-positive samples were rescreened using a fluorescence polarization immunoassay (FPIA). Specimens positive by both EMIT and FPIA were confirmed by gas chromatography/mass spectrometry (GC/MS) if sufficient quantity of urine was available. BE was identified in 41 (4.4%) and opiates in 46 (4.9%) patients by both EMIT and FPIA. The presence of BE or opiate was confirmed by GC/MS in all 34 cases where sufficient urine was available. The age- and sex-adjusted systolic and diastolic blood pressure percentiles were greater, and head circumference and weight percentiles were lower in BE-positive patients compared to those with negative drug screens. There were no associations between opiate exposure and any of these variables. We conclude that occult postnatal cocaine exposure is associated with measurable physical and physiologic differences.


[Nodular hepatic lesions secondary to multiple myeloma].
Multiple myeloma is characterized by the neoplastic proliferation of a single clone of plasma cells producing a monoclonal immunoglobulin. Myelomatous nodular lesions of the liver are infrequent. We describe 3 cases with histological confirmation and we review the bibliography.


Modeling the influence of attitudes, trust, and beliefs on endoscopists' acceptance of artificial intelligence applications in medical practice.
The potential for deployment of Artificial Intelligence (AI) technologies in various fields of medicine is vast, yet acceptance of AI amongst clinicians has been patchy. This research therefore examines the role of antecedents, namely trust, attitude, and beliefs in driving AI acceptance in clinical practice. We utilized online surveys to gather data from clinicians in the field of gastroenterology. A total of 164 participants responded to the survey. Participants had a mean age of 44.49 (SD = 9.65). Most participants were male (n = 116, 70.30%) and specialized in gastroenterology (n = 153, 92.73%). Based on the results collected, we proposed and tested a model of AI acceptance in medical practice. Our findings showed that while the proposed drivers had a positive impact on AI tools' acceptance, not all effects were direct. Trust and belief were found to fully mediate the effects of attitude on AI acceptance by clinicians. The role of trust and beliefs as primary mediators of the acceptance of AI in medical practice suggest that these should be areas of focus in AI education, engagement and training. This has implications for how AI systems can gain greater clinician acceptance to engender greater trust and adoption amongst public health systems and professional networks which in turn would impact how populations interface with AI. Implications for policy and practice, as well as future research in this nascent field, are discussed.


Dose-response effects of the text4baby mobile health program: randomized controlled trial.
Mobile health (mHealth) is growing rapidly, but more studies are needed on how to optimize programs, including optimal timing of messaging, dose of exposure, and value of interactive features. This study evaluates final outcomes of text4baby (a text message service for pregnant and postpartum women) from a randomized trial performed in a population of pregnant female soldiers and family members. The study aims were to evaluate (1) treatment effects and (2) dose-response effects of text4baby on behavioral outcomes compared to control (no text4baby) condition. The study was a randomized trial of text4baby at Madigan Army Medical Center. Female military health beneficiaries who met inclusion criteria were eligible for the study. Participants provided consent, completed a baseline questionnaire, and then were randomized to enroll in text4baby or not. They were followed up at 3 time points thereafter through delivery of their baby. Generalized estimating equation models were used to evaluate outcomes. We examined treatment effects and the effects of higher doses of text4baby messages on outcomes. We report descriptive statistics including dosage of text messages delivered. The main finding was a significant effect of high exposure to text4baby on self-reported alcohol consumption postpartum (OR 0.212, 95% CI 0.046-0.973, P=.046), as measured by the question "Since you found out about your pregnancy, have you consumed alcoholic beverages?" The text4baby participants also reported lower quantities of alcohol consumed postpartum. Studies of text4baby have helped to build the mHealth evidence base. The effects of text4baby offer lessons for future scalable mHealth programs and suggest the need to study dose-response effects of these interventions.


Door-to-balloon times under 90 min can be routinely achieved for patients transferred for ST-segment elevation myocardial infarction percutaneous coronary intervention in a rural setting.
The purpose of this study was to demonstrate the feasibility of routine transfer of ST-segment elevation myocardial infarction (STEMI) patients to achieve percutaneous coronary intervention (PCI) in less than 90 min from presentation. Many PCI hospitals have achieved routine door-to-balloon times under 90 min for patients with STEMI presenting directly to the hospital. However, few patients transferred from a non-PCI center undergo PCI within 90 min of presentation. Our rural PCI hospital implemented a program in 2005 for rapid triage, transfer, and treatment of STEMI patients and made additional improvements in 2006 and 2007. Intervals between milestones in the STEMI triage/transfer/treatment process were assessed before and after implementation of the program. During the 5-year study period, 676 patients with 687 STEMIs were transferred from 19 community hospitals and underwent PCI. Median door-to-balloon time decreased from 189 min to 88 min (p < 0.001). The time intervals reflecting efficiency of the referring hospitals, transfer services, and PCI hospital all significantly improved. In 2008, median door-to-balloon times were <90 min for 6 of the 7 most frequently referring hospitals. Delays during off-hours presentation in 2004 were abolished after the program was implemented in 2005. In-hospital mortality decreased from 6% before to 3% after implementation of the program. In multivariate modeling, presentation before initiation of the STEMI program predicted increased risk of in-hospital mortality (odds ratio: 3.74, 95% confidence interval: 1.22 to 11.51, p = 0.021). A program of rapid triage, transfer, and treatment of STEMI patients presenting to non-PCI hospitals can reduce in-hospital mortality and produce progressive improvements in door-to-balloon time such that median door-to-balloon times under 90 min are feasible.


Mepolizumab rescue therapy for acute pneumonitis secondary to DRESS.
Drug reaction with eosinophilia and systemic symptoms (DRESS) syndrome represents a severe adverse drug reaction driven by eosinophilia. Treatment is focused on withdrawal of medication, supportive care and immunosuppression such as high-dose corticosteroid therapy. Here we report a 56-year-old male patient who initially presented with breathlessness and eosinophilia, subsequent development of respiratory failure and admission to ITU for non-invasive ventilation. The patient continued to deteriorate despite high-dose prednisolone and methylprednisolone. Other causes of hypereosinophilia were normal. He was diagnosed with DRESS syndrome secondary to pregabalin and was treated with subcutaneous mepolizumab. We observed the rapid resolution of eosinophilia and clinical improvement; the patient was discharged home within a month of administration. This represents the successful use of mepolizumab in the acute setting of pulmonary failure secondary to DRESS. A similar approach could be adopted in other acute conditions with refractory eosinophilic inflammation where standard steroid therapy has failed.


[Successful Treatment of Blow-out Type Left Ventricular Free Wall Rupture Using Sutureless Technique].
Blow-out type left ventricular free wall rupture is a serious complication of acute myocardial infarction, that carries high hospital mortality rates and poor surgical outcome. We report the case of an 88-year-old woman who developed cardiac tamponade following percutaneous coronary angioplasty for acute myocardial infarction. She was diagnosed with left ventricular free wall rupture, and rupture type was proved to be blow out after median sternotomy. To address this critical condition, we opted for the sutureless technique for its minimally invasive nature and ability to preserve left ventricular function. The patient was discharged from the hospital without any complications 22 days after surgery. Considering favorable, encouraging outcomes of this case, sutureless technique could be regarded as a viable option for blow-out type left ventricular free wall rupture.


Geographical Distribution, Incidence, Malignancies, and Outcome of 136 Eastern Slavic Patients With Nijmegen Breakage Syndrome and NBN Founder Variant c.657_661del5.
Nijmegen breakage syndrome (NBS) is a DNA repair disorder characterized by combined immunodeficiency and a high predisposition to lymphoid malignancies. The majority of NBS patients are identified with a homozygous five base pair deletion in the Nibrin (NBN) gene (c.657_661del5, p.K219fsX19) with a founder effect observed in Caucasian European populations, especially of Slavic origin. We present here an analysis of a cohort of 136 NBS patients of Eastern Slav origin across Belarus, Ukraine, Russia, and Latvia with a focus on understanding the geographic distribution, incidence of malignancy, and treatment outcomes of this cohort. Our analysis shows that Belarus had the highest prevalence of NBS (2.3 per 1,000,000), followed by Ukraine (1.3 per 1,000,000), and Russia (0.7 per 1,000,000). Of note, the highest concentration of NBS cases was observed in the western regions of Belarus and Ukraine, where NBS prevalence exceeds 20 cases per 1,000,000 people, suggesting the presence of an "Eastern Slavic NBS hot spot." The median age at diagnosis of this cohort ranged from 4 to 5 years, and delay in diagnosis was more pervasive in smaller cities and rural regions. A total of 62 (45%) patients developed malignancies, more commonly in males than females (55.2 vs. 34.2%; p=0.017). In 27 patients, NBS was diagnosed following the onset of malignancies (mean age: 8 years). Malignancies were mostly of lymphoid origin and predominantly non-Hodgkin lymphoma (NHL) (n=42, 68%); 38% of patients had diffuse large B-cell lymphoma. The 20-year overall survival rate of patients with malignancy was 24%. However, females with cancer experienced poorer event-free survival rates than males (16.6% vs. 46.8%, p=0.036). Of 136 NBS patients, 13 underwent hematopoietic stem cell transplantation (HSCT) with an overall survival of 3.5 years following treatment (range: 1 to 14 years). Indications for HSCT included malignancy (n=7) and immunodeficiency (n=6). Overall, 9% of patients in this cohort reached adulthood. Adult survivors reported diminished quality of life with significant physical and cognitive impairments. Our study highlights the need to improve timely diagnosis and clinical management of NBS among Eastern Slavs. Genetic counseling and screening should be offered to individuals with a family history of NBS, especially in hot spot regions.


Multi-animal pose estimation, identification and tracking with DeepLabCut.
Estimating the pose of multiple animals is a challenging computer vision problem: frequent interactions cause occlusions and complicate the association of detected keypoints to the correct individuals, as well as having highly similar looking animals that interact more closely than in typical multi-human scenarios. To take up this challenge, we build on DeepLabCut, an open-source pose estimation toolbox, and provide high-performance animal assembly and tracking-features required for multi-animal scenarios. Furthermore, we integrate the ability to predict an animal's identity to assist tracking (in case of occlusions). We illustrate the power of this framework with four datasets varying in complexity, which we release to serve as a benchmark for future algorithm development.


Evaluation of amniotic fluid in preterm labor with intact membranes.
Amniotic fluid was obtained from 35 pregnant women in preterm labor with intact membranes. Their gestational ages ranged from 24 to 34 weeks. Bacteria were detected in only 1 (3%) of the 35 amniotic fluid samples. The anaerobic culture grew Bacteroides corrodens and Fusobacterium nucleatum. The L/S ratio was greater than 4.0 in six patients (17%), 2.5-4.0 in two (6%) and less than 2.5 in 23 (66%); the quantity of fluid was inadequate for L/S analysis in four (11%). None of the 35 newborns developed evidence of infection in the neonatal period. Only two women (6%) were febrile postpartum, and none experienced a prolonged hospital stay. Seven (20%) of the infants developed respiratory distress syndrome. Five (14%) developed hyaline membrane disease, and two (6%) had transient tachypnea of the newborn. Intrauterine infection may play a lesser role in preterm labor with intact membranes than previously postulated. Amniocentesis may provide useful information for directing management by permitting one to assess fetal pulmonary maturity and the presence of meconium.


Endothelin-1 causes P-selectin-dependent leukocyte rolling and adhesion within rat mesenteric microvessels.
Endothelin-1 (ET-1) is a potent vasoconstrictor postulated to play a role in hypertension, ischemia-reperfusion, and atherosclerosis. In addition to these contributions, it has been also proposed to induce leukocyte-endothelial cell interactions. The aim of the present study was to assess the mechanisms of action of ET-1 on leukocyte recruitment in vivo. Intravital microscopy of the rat mesenteric postcapillary venules was used. Ten minutes after 1 nM ET-1 superfusion, a significant increase in leukocyte rolling (77.5 +/- 22.6 vs. 20.5 +/- 4.5 cells/min) and adhesion (15.5 +/- 2.9 vs. 3.0 +/- 0.8 cells/100 micrometer) but not emigration was observed. These effects were found not to be mediated by mast cell activation. No platelet-endothelial cell interactions were detected in this in vivo system and furthermore, flow cytometry analysis revealed no increase of P-selectin expression in rat platelets on ET-1 stimulation. Pretreatment of animals with an anti-rat P-selectin monoclonal antibody (mAb) dramatically reduced leukocyte rolling and adhesion by 100 and 94% respectively when compared with control mAb-treated animals. At this dose of ET-1, a very transient decrease in shear rate was detected, arteriolar diameter was significantly reduced but venular diameter remained unchanged. A similar mechanical reduction in blood flow did not induce leukocyte recruitment. Thus this study demonstrates that ET-1 can directly cause significant leukocyte rolling and adhesion adding to its potential pathophysiological role in the development of disease states of the cardiovascular system.


Does Treatment with Dexmedetomidine Intra-articularly Improve Postoperative Pain and Rehabilitation after Anterior Cruciate Ligament Reconstruction?
This study aims to evaluate the analgesic efficacy of dexmedetomidine added to levobupivacaine following anterior cruciate ligament (ACL) surgery. Fifty patients undergoing ACL reconstruction were included. Group DL (dexmedetomidin-levobupivacaine) received 20 mL 0.5% levobupivacaine plus 1 mL (100 μg) dexmedetomidine. Group L (levobupivacaine) patients received 20 mL 0.5% levobupivacaine plus 1 mL saline 10 minutes before tourniquet release. A patient-controlled analgesia (PCA) pump was then connected, delivering 0.5 mg at every 10 minutes and 1-mg morphine and 75-mg diclofenac sodium was used as a rescue analgesic. Postoperative pain was evaluated 0, 2, 4, 6, 12, and 24 hours after extubation at rest and during movement. A rehabilitation program was started after surgery. Postoperative continuous passive motion (CPM) starting time, postoperative leg flexion angle, and straight leg lifting time were evaluated for each group. There were no significant differences between the groups in terms of demographic data and operation time. Morphine consumption, analgesic requirements, and visual analogue scale (VAS) assessments were significantly lower in group DL during the 24-hour period after surgery. The time to start CPM in the postoperative period was significantly shorter in group DL. Passive joint flexion angle was significantly higher in group DL. Postoperative straight leg lifting time was significantly shorter in group DL. Adding dexmedetomidine to the intra-articular levobupivacaine provided better postoperative pain control and improved rehabilitation period after ACL surgery.


Generation of virtual monochromatic CBCT from dual kV∕MV beam projections.
To develop a novel on-board imaging technique which allows generation of virtual monochromatic (VM) cone-beam CT (CBCT) with a selected energy from combined kilovoltage (kV)∕megavoltage (MV) beam projections. With the current orthogonal kV∕MV imaging hardware equipped in modern linear accelerators, both MV projections (from gantry angle of 0°-100°) and kV projections (90°-200°) were acquired as gantry rotated a total of 110°. A selected range of overlap projections between 90° to 100° were then decomposed into two material projections using experimentally determined parameters from orthogonally stacked aluminum and acrylic step-wedges. Given attenuation coefficients of aluminum and acrylic at a predetermined energy, one set of VM projections could be synthesized from two corresponding sets of decomposed projections. Two linear functions were generated using projection information at overlap angles to convert kV and MV projections at nonoverlap angles to approximate VM projections for CBCT reconstruction. The contrast-to-noise ratios (CNRs) were calculated for different inserts in VM CBCTs of a CatPhan phantom with various selected energies and compared with those in kV and MV CBCTs. The effect of overlap projection number on CNR was evaluated. Additionally, the effect of beam orientation was studied by scanning the CatPhan sandwiched with two 5 cm solid-water phantoms on both lateral sides and an electronic density phantom with two metal bolt inserts. Proper selection of VM energy [30 and 40 keV for low-density polyethylene (LDPE), polymethylpentene, 2 MeV for Delrin] provided comparable or even better CNR results as compared with kV or MV CBCT. An increased number of overlap kV and MV projection demonstrated only marginal improvements of CNR for different inserts (with the exception of LDPE) and therefore one projection overlap was found to be sufficient for the CatPhan study. It was also evident that the optimal CBCT image quality was achieved when MV beams penetrated through the heavy attenuation direction of the object. A novel technique was developed to generate VM CBCTs from kV∕MV projections. This technique has the potential to improve CNR at selected VM energies and to suppress artifacts at appropriate beam orientations.


Genetic structure and diversity of native Guadua species (Poaceae: Bambusoideae) in natural populations of the Brazilian Amazon rainforest.
The Southwestern Region of the Brazilian Amazon is formed by forests dominated by bamboos. The genus Guadua is endemic to the Americas, and little is known about the genetic diversity and structure of species of this genus. This study aimed to evaluate the genetic diversity and structure of two native Guadua species in natural populations in the Southwestern region of the Brazilian Amazon. Therefore, the genetic diversity and structure of Guadua aff. chaparensis and Guadua aff. lynnclarkiae were evaluated with the use of microsatellite molecular markers (SSR). It was verified that the average genetic diversity for the populations studied was considered high ( H ^ e =0.5) compared to other species of bamboo. All populations had rare and private alleles, and none of them presented significant values of inbreeding. The populations were divergent ( G ^ ST = 0.46), resulting in a low apparent gene flow. The Bayesian analysis showed that among the 350 individuals analyzed, five groups (K=5) were formed, with little similarity among the groups (Populations), although two of them presented clonal individuals. According to the results obtained, it can be conclude that populations should be treated as having unique characteristics, mainly when accessed for management and for in situ and ex situ conservation studies.


Effect of different carbon sources on proteases secreted by the fungal pathogen Sclerotinia sclerotiorum during Phaseolus vulgaris infection.
Sclerotinia sclerotiorum (Sclerotiniaceae) is a plant pathogenic fungus that causes white mold disease in vegetable crops, including the common bean (Phaseolus vulgaris). Proteases produced by fungi are normally an important part of the pathogenic process in the host. We examined the effect of different carbon sources--pectin, glucose, and cell wall of P. vulgaris on the production of proteases in cultures of S. sclerotiorum. These proteases were also assayed in infected P. vulgaris plants. Enzyme activity was increased with all carbon sources, but the highest levels were found when pectin was added. Based on real-time quantitative reverse transcription-PCR analyses, protease induction in S. sclerotiorum was found to occur at the level of gene transcription. The finding of increased expression of acid phosphatase 1 and aspartyl protease in vivo in infected P. vulgaris plants supports the role of these enzymes in the invasion process of S. sclerotiorum.


Impact of wellness indicators on intellectual dimensions of medical teachers of Karachi: Findings from cross sectional study.
To measure impact of social, spiritual, occupational, physical, financial and emotional wellness on intellectual wellness of medical teachers. The cross-sectional study was conducted at 8 medical universities in the Sindh province of Pakistan from January 2012 to December 2014, and comprised faculty members. Using a self-administered questionnaire, teachers' responses on physical, emotional, social, spiritual, intellectual, occupational and financial spokes of the wellness wheel that were graded on a 4-point Likert scale. Generalised linear models were executed to find the best predictive scores for intellectual wellness. Out of 300 subjects approached, 261(87%) returned the questionnaire fully completed. Of them, 86(33%) were males and 175(67%) were females. Occupational wellness followed by spiritual, financial and emotional wellness depicted significantly high correlation with IW (p<0.05). Adjusted analysis revealed significant effects of spiritual, financial and occupational wellness (p<0.05). Findings of Models 1 and 2 suggested that holding the effect of other wellness subscales, intellectual wellness increased up to 0.485 score and 0.245 score due to occupational and spiritual wellness (p<0.05) respectively, while financial wellness was not significant (p>0.05). In the final model, occupational wellness effect increased to 0.504 score and spiritual wellness effect projected to 0.273 score for intellectual wellness (p<0.05). Amongst all the spokes of the wellness wheel, occupational, financial, social, emotional and spiritual wellness improved mental capabilities of medical teachers to overcome the challenges for acquiring intellectual wellness. However, wellness at workplace played the major role in enhancing intellectual proficiencies.


Erythrovirus B19 and autoimmune thyroid diseases. Review of the literature and pathophysiological hypotheses.
Erythrovirus B19 (EVB19) has been incriminated, over recent years, in the onset and/or pathogenesis of many diseases, especially autoimmune thyroid diseases. This review of the literature (published over the last 40 years using Pubmed and Science Direct search engines) was designed to define the role of EVB19, particularly in autoimmune thyroid diseases.Two cases of subacute thyroiditis, one case of Graves' disease (associated with type 1 diabetes and rheumatoid arthritis), and one case of Hashimoto's thyroiditis following acute EVB19 infection were reported. A retrospective case-control study in a pediatric population demonstrated the role of EVB19 in Hashimoto's thyroiditis. Four retrospective studies of pathology slides (including PCR, immunohistochemistry or in situ hybridization) and a prospective case-control study on pathology slides demonstrated the presence of EVB19 in thyroid tissue of patients with benign multinodular goiter, Graves' disease, autoimmune thyroiditis (including Hashimoto's thyroiditis), and thyroid cancer. EVB19 can be demonstrated in the thyroid gland in a wide range of diseases. Although acute EVB19 infection could theoretically trigger autoimmune thyroid disease, there is currently no evidence that EVB19 plays a specific role in the pathophysiology of autoimmune thyroid diseases.


Impurity profile study of dutasteride.
During the process development of dutasteride in the laboratory analysis showed some impurity peaks in HPLC ranging from 0.05 to 0.1%. The same samples were analyzed by LCMS method and identified peak at m/z 508 (desmethyl dutasteride), 530 (dihydro dutasteride) and 528 (isomer of dutasteride). These impurities were synthesized individually and characterized based on the spectroscopic data (HPLC, IR, NMR and MS). The structures of these impurities were 17beta-N-[2,5-bis(trifluoromethyl) phenyl]carbamoyl-3-hydroxyl-4-azaestra-1,3,5,7,9-pentaene (desmethyl of dutasteride 2), 17beta-N-[2,5-bis(trifluoromethyl)phenyl]carbamoyl-4-aza-5alpha-androstane-3-one (dihydro impurity of dutasteride 3), and 17beta-N-[2,5-bis(trifluoromethyl) phenyl] carbamoyl-4-aza-5beta-androst-1-ene-3-one (beta-isomer of dutasteride 4), respectively. The formation and synthesis of dutasteride impurities are discussed.


Motor neuron regeneration in adult zebrafish.
The mammalian spinal cord does not regenerate motor neurons that are lost as a result of injury or disease. Here we demonstrate that adult zebrafish, which show functional spinal cord regeneration, are capable of motor neuron regeneration. After a spinal lesion, the ventricular zone shows a widespread increase in proliferation, including slowly proliferating olig2-positive (olig2+) ependymo-radial glial progenitor cells. Lineage tracing in olig2:green fluorescent protein transgenic fish indicates that these cells switch from a gliogenic phenotype to motor neuron production. Numbers of undifferentiated small HB9+ and islet-1+ motor neurons, which are double labeled with the proliferation marker 5-bromo-2-deoxyuridine (BrdU), are transiently strongly increased in the lesioned spinal cord. Large differentiated motor neurons, which are lost after a lesion, reappear at 6-8 weeks after lesion, and we detected ChAT+/BrdU+ motor neurons that were covered by contacts immunopositive for the synaptic marker SV2. These observations suggest that, after a lesion, plasticity of olig2+ progenitor cells may allow them to generate motor neurons, some of which exhibit markers for terminal differentiation and integration into the existing adult spinal circuitry.


Effectiveness of a quit and win contest with a low-income population.
In a two-group quasi-experimental study, we evaluated the impact of a quit and win contest on quitting among low-income tobacco users and identified contest elements used by successful quitters. Low-income tobacco users have been largely untouched by tobacco cessation approaches. A volunteer sample of 248 low-income tobacco users were recruited from quit and win contest registrants (treatment group). A random sample of 290 low-income tobacco users who had not entered the contest were recruited using random digit dialing (control group). Telephone interviews were conducted with both groups at baseline, 3, 6, and 12 months. Seven-day point prevalence measured self-reported quitting and urine cotinine assessed confirmed quitting. On average, quit and win study participants were 3.5 times more likely than controls to self-report quitting and 12.8 times more likely to demonstrate confirmed quitting after controlling for baseline differences in stage of change, age, education, and marital status. The use of specific contest elements was not related to successful quitting. The overall quit rates in the treatment group were higher than those in the control group. The results are promising given that low-income tobacco users are generally less likely to succeed in quitting.


LncNORAD interference inhibits tumor growth and lung cancer cell proliferation, invasion and migration by down-regulating CXCR4 to suppress RhoA/ROCK signaling pathway.
Non-small cell lung cancer (NSCLC) accounts for the majority of lung cancer, with an unfavorable prognosis of 5-year survival rates. It is of great clinical significance to further search for more efficacious and novel targets for diagnosis and therapeutic strategies. This study aimed at clarifying the role of long non-coding RNA (lncRNA) NORAD in proliferation, invasion and migration and tumor growth of NSCLC. In this study, mRNA levels of lncRNA NORAD were examined by RT-PCR. CCK-8 assay was applied to test cell viability. Furthermore, wound healing assay and transwell assay were performed to detect the migration and invasion of A549 cells, respectively. Immunohistochemistry was applied to assess the levels of CXC chemokine receptor (CXCR) 4 and CXC chemokine ligand (CXCL) 12. Mice models of NSCLC in vivo were exploited to further examine the potential role of NORAD in tumor growth. Key proteins related to Ras homolog gene family member A (RhoA) GTPase/Rho-associated kinase (RhoA/ROCK) pathway were determined by Western blot. NORAD has elevated the levels in NSCLC tissues and cells. NORAD interference dramatically inhibited tumor growth and suppressed A549 cell proliferation, migration and invasion by downregulating CXCR4 and CXCL12 expression. RhoA/ROCK signaling pathway was activated in NSCLC. This study revealed that the downregulation of lncRNA NORAD could slow down the progression of NSCLC by regulating CXCR4 and RhoA/ROCK signaling pathway.


The effects of high-pass and of low-pass filtering upon the intelligibility of speech in white noise.
This paper studied with trained, normal-hearing, young adults the effects of high-pass and also low-pass filtering on the intelligibility of speech mixed at S's ear with additive white noise at 4 S/N. Before each filter--S/N combination Ss were encouraged to adjust overall gain so as to yield seemingly optimum speech intelligibility. Gain chosen was relatively independent of S/N, being primarily a function of speech level. Results provided information on the contribution to intelligibility of various frequency ranges of the speech spectrum. Data were examined in relation also to the articulation index (AI). The intelligibility scores were in good agreement with the AI and yielded an improved understanding of the intelligibility of speech in noise.


Which Components of Metabolic Syndrome have a Greater Effect on Mortality, CVA and Myocardial Infarction, Hyperglycemia, High Blood Pressure or Both?
This study aims to evaluate the incidence of stroke, ischemic heart disease (IHD), and cardiovascular disease (CVD) mortality in clusters containing hypertension (HTN), clusters containing diabetes (diabetes mellitus [DM]), cluster with both HTN, DM, and cluster without HTN, DM in patients with metabolic syndrome (MetS). The association between MetS and outcomes was examined in 2257 patients with MetS that were divided into four groups includes patients with MetS with hyperglycemia (Cluster 1), patients with MetS with HTN (Cluster 2), patients with MetS with HTN and hyperglycemia (Cluster 3) and patients with MetS without HTN and hyperglycemia (Cluster 4). To assess the risk ratio and incidence of CVA, myocardial infarction, and mortality with the use multivariate Cox proportional hazards models. As it shown the lowest prevalence of events was in Cluster 1 and against in Cluster 3 the prevalence of stroke with 34 (38.2%) cases and the prevalence of IHD and CVD mortality in Cluster 2 with, respectively, 168 (54.7%) and 51 patients (49%) had the most frequencies (P < 0.001), and hence that the lowest prevalence of events was seen in Cluster 1, but stroke in Cluster 3 with 34 cases (38.2%) and the prevalence of IHD and CVD mortality in Cluster 2, respectively, with 168 (54.7%) and 51 patients (49%) had the most frequencies (P < 0.001). More intensive changes in lifestyle and management protocols may be required in these patients for controlling the components of the syndrome, with the aim of preventing not only type II diabetes and CVD but also ischemic cerebrovascular events.


Are Androgen and Estrogen Receptors in DCIS Patients Prognostic Indicators of Relapse Independently of Treatment?
Ductal carcinoma in situ (DCIS) is a highly heterogenous tumor that is now more frequently diagnosed because of the increased number of screening programs. Women with DCIS are mainly treated with conservative surgery almost always followed by radiotherapy. Although conventional biomarkers, i.e. ER, PgR, Ki67, and HER2, have been extensively investigated in invasive tumors, little is known about their role in DCIS, especially that of the androgen receptor (AR). In the present study, the expression of conventional biomarkers and AR was determined by immunohistochemistry in 85 DCIS samples from patients monitored for a maximum of 13 years: 43 patients were treated with quadrantectomy and 42 patients underwent quadrantectomy and radiotherapy. Of these, 5 and 11 patients relapsed, respectively. Our findings showed that ER and PgR were higher in nonrelapsed than in relapsed patients (P=0.025 and 0.0038). In contrast, AR expression and the AR/ER ratio were higher in relapsed patients than in the nonrelapsed group (P=0.0069 and 0.0012). At the best cut-off value of 1.1, the AR/ER ratio showed an overall accuracy of 92% and 80% in predicting in situ relapse or progression to invasive carcinoma in DCIS patients treated with surgery and those treated with surgery plus radiotherapy, respectively. AR would therefore appear to be an independent prognostic biomarker in the latter group. Our preliminary results highlight the potentially important role of the AR/ER ratio as a predictive indicator of DCIS relapse, independently of treatment.


Pseudovirion-binding and neutralizing antibodies to cutaneous human papillomaviruses (HPV) correlated with the presence of HPV DNA in skin.
Whereas the antibody response to the anogenital human papillomaviruses (HPVs) is known to be mainly type-specific, correlated with the presence of viral DNA and mainly directed to conformational epitopes of the virion, it is not known if this applies also to the antibody response to cutaneous HPVs. For 434 non-immunosuppressed patients with skin lesions (squamous cell carcinoma and basal cell carcinoma of the skin, actinic keratosis and benign skin lesions), we compared HPV DNA status with seroreactivity to HPV pseudovirions (PsV) and to GST-L1 fusion proteins from HPV types -5, -6, -15, -16, -32 and -38. Biopsies from the skin lesions were tested for the presence of HPV DNA using three different PCR methods, with typing by sequencing. Serum samples from subjects with HPV DNA-positive biopsies and randomly selected serum samples from subjects with HPV DNA-negative biopsies were also tested with neutralization assays with HPV5, -38 and -76 PsV. Agreement of the three serological methods varied from poor to moderate. Type-specific seroprevalences among patients positive for the same type of HPV DNA (sensitivity of serology) was improved with the PsV-based method (mean of 40%, maximum 63%) compared with the GST-L1 method (mean of 20%, maximum of 25%). Neutralization was the most sensitive assay for HPV38 (50%). In summary, cutaneous HPVs also appear to induce a type-specific antibody response that correlates with the presence of HPV DNA and that can be detected with improved sensitivity using PsV-based serology.


Noninvasive mechanical ventilation with average volume assured pressure support (AVAPS) in patients with chronic obstructive pulmonary disease and hypercapnic encephalopathy.
Non-invasive mechanical ventilation (NIV) in patients with acute respiratory failure has been traditionally determined based on clinical assessment and changes in blood gases, with NIV support pressures manually adjusted by an operator. Bilevel positive airway pressure-spontaneous/timed (BiPAP S/T) with average volume assured pressure support (AVAPS) uses a fixed tidal volume that automatically adjusts to a patient's needs. Our study assessed the use of BiPAP S/T with AVAPS in patients with chronic obstructive pulmonary disease (COPD) and hypercapnic encephalopathy as compared to BiPAP S/T alone, upon immediate arrival in the Emergency-ICU. We carried out a prospective interventional match-controlled study in Guayaquil, Ecuador. A total of 22 patients were analyzed. Eleven with COPD exacerbations and hypercapnic encephalopathy with a Glasgow Coma Scale (GCS) <10 and a pH of 7.25-7.35 were assigned to receive NIV via BiPAP S/T with AVAPS. Eleven patients were selected as paired controls for the initial group by physicians who were unfamiliar with our study, and these patients were administered BiPAP S/T. Arterial blood gases, GCS, vital signs, and ventilatory parameters were then measured and compared between the two groups. We observed statistically significant differences in favor of the BiPAP S/T + AVAPS group in GCS (P = .00001), pCO(2) (P = .03) and maximum inspiratory positive airway pressure (IPAP) (P = .005), among others. However, no significant differences in terms of length of stay or days on NIV were observed. BiPAP S/T with AVAPS facilitates rapid recovery of consciousness when compared to traditional BiPAP S/T in patients with chronic obstructive pulmonary disease and hypercapnic encephalopathy. Current Controlled Trials application ref is ISRCTN05135218.


[Parameters of membrane permeability, microcirculation, external respiration, and trace element levels in the drug-laser treatment of pneumonia].
To study effects of laser therapy in combined treatment of pneumonia on external respiration function, membrane permeability, microcirculation and serum trace elements. 142 pneumonia patients were randomly divided into two groups: 96 patients treated with drugs and laser radiation (the study group) and 46 patients treated with drugs only (control group). In the study group there was more pronounced reduction in cell membrane permeability, a rise in concentrations of iron and chromium in the blood serum, improvement of microcirculation. These changes closely correlated with those in immunity, external respiration function. Laser therapy is an effective method of pneumonia treatment and can be included in relevant combined schemes.


Mutations in CFAP43 and CFAP44 cause male infertility and flagellum defects in Trypanosoma and human.
Spermatogenesis defects concern millions of men worldwide, yet the vast majority remains undiagnosed. Here we report men with primary infertility due to multiple morphological abnormalities of the sperm flagella with severe disorganization of the sperm axoneme, a microtubule-based structure highly conserved throughout evolution. Whole-exome sequencing was performed on 78 patients allowing the identification of 22 men with bi-allelic mutations in DNAH1 (n = 6), CFAP43 (n = 10), and CFAP44 (n = 6). CRISPR/Cas9 created homozygous CFAP43/44 male mice that were infertile and presented severe flagellar defects confirming the human genetic results. Immunoelectron and stimulated-emission-depletion microscopy performed on CFAP43 and CFAP44 orthologs in Trypanosoma brucei evidenced that both proteins are located between the doublet microtubules 5 and 6 and the paraflagellar rod. Overall, we demonstrate that CFAP43 and CFAP44 have a similar structure with a unique axonemal localization and are necessary to produce functional flagella in species ranging from Trypanosoma to human.


Fatal parasite-induced enteritis and typhlocolitis in horses in Southern Brazil.
Diseases related to the alimentary system are the main cause of death in horses. This retrospective study aimed to describe the pathological findings of fatal parasite-induced enteritis and typhlocolitis caused by cyathostominae, Eimeria leuckarti, Balantidium coli, and Strongyloides westeri in horses. The records of parasite-induced intestinal lesions in horses necropsied in Southern Brazil between 2005 and 2017 were reviewed. Ten horses had fatal parasitic enteritis and/or typhlocolitis, and the main causes were: cyathostominae typhlocolitis (6/10), E. leuckarti enteritis (1/10), S. westeri enteritis (1/10), B. coli colitis related to cyathostominae (1/10), and infection by multiple agents (1/10). Cyathostominae typhlocolitis showed marked mucosal thickening, with multifocal elevated nodules containing tangled filiform parasites. Microscopic examination revealed that the mucosa and submucosa had encysted parasitic structures surrounded by eosinophilic and granulomatous inflammation. E. leuckarti enteritis was microscopically characterized by macrogamonts, microgamonts, and oocysts inside the host cells. S. westeri enteritis showed microscopic atrophy of the villi with numerous mucosal encysted parasitic structures. B. coli typhlocolitis showed severe diffuse mucosal reddening, with microscopic superficial mucosal necrosis associated with multiple protozoan trophozoites. Fatal parasite-induced enteritis and typhlocolitis are important causes of death in horses in Southern Brazil.


Degradation of adsorbed fibrinogen by surface-generated plasmin.
An investigation of the physical status of fibrinogen that has been adsorbed to glass and then eluted has been conducted. Exposure of Kabi fibrinogen to glass was carried out using a glass bead column experiment. The fibrinogen was eluted sequentially, first by 1 M Tris and then by SDS. The initially eluted fibrinogen showed considerable degradation (SDS polyacrylamide gel electrophoresis) while the later fractions were less degraded. Fibrinogen purified by chromatography on either DEAE-cellulose or Sepharose-lysine to remove plasminogen was less degraded. When purified plasminogen was added to fibrinogen, degradation of column eluates was very extensive in all eluted fractions. These results are interpreted in terms of a surface-mediated activation of plasminogen to plasmin followed by fibrinogenolysis. Although such an effect remains to be demonstrated as a general property of surfaces, it is speculated that surfaces may vary in their activation of fibrinolysis as well as their activation of clotting, and that maximization of fibrinolysis is a worthwhile goal in the development of blood-compatible surfaces.


PTPRC functions as a prognosis biomarker in the tumor microenvironment of cutaneous melanoma.
Cutaneous melanoma is one of the most malignant types of skin cancer, with an extremely poor prognosis. Immune cells infiltrated in the tumor microenvironment (TME) affects melanoma initiation, progression, prognosis and immunotherapy strategies in melanoma. The potential utility of TME-related genes as a prognostic model for melanoma and as a predictor of immunotherapeutic response merits further exploration. In this study, we determined that an immune-related gene, protein tyrosine phosphatase receptor type C (PTPRC), was positively correlated with the positive prognosis of melanoma patients. Integration of this gene with TNM classification created a predictive model that showed better performance in determining overall survival than others. PTPRC expression was positively correlated with the levels of immune checkpoint molecules, and PTPRC knockdown significantly enhanced the migration, invasion, and proliferation of melanoma cells. Finally, immunohistochemical results from HPA and Real-time quantitative PCR of clinical tissues confirmed that PTPRC expression was higher in melanoma than in normal skin. In conclusion, PTPRC served as a potential predictor of survival and response to immunotherapy in melanoma patients. The risk model combining the PTPRC and TNM classifications holds the potential to be a promising tool for prognostic prediction of cutaneous melanoma. This will help in the effective clinical management of melanoma patients.


Polytetrafluoroethylene graft aneurysms: a report of five aneurysms.
True aneurysms have developed in nonreinforced, expanded polytetrafluoroethylene (PTFE). We report our experience in arterial reconstructions with PTFE in the legs of patients having symptomatic occlusive disease. Fifty-four femoropopliteal or femorotibial bypass operations were performed in 51 patients between January 1975 and December 1976. An unsuitable saphenous vein for a vascular conduit was found in 14 patients, and reconstruction was done with PTFE (IMPRA-graft). Two of the 14 PTFE grafts were composite and used a segment of saphenous vein. Aneurysmal dilation developed in five PTFE grafts (36%) in four patients three to 36 months following placement. Nonreinforced PTFE conduits as manufactured during 1975 and 1976 are unsatisfactory for use as limb arterial bypass grafts because of the high incidence of aneurysmal change. Those that are implanted must be followed up indefinitely because of late aneurysmal formation and rupture. Newer PTFE graft modifications should be used with caution.


COPD-Related Anxiety: A Systematic Review of Patient Perspectives.
Anxiety in patients with chronic obstructive pulmonary disease (COPD) is prevalent but often unidentified and therefore not adequately managed. Clinicians find it difficult to detect anxiety symptoms and to differentiate subclinical anxiety from anxiety disorders, because of the considerable overlap between symptoms of COPD and anxiety. We synthesized existing qualitative research on patients' experiences of COPD-related anxiety with the purpose of gaining a richer understanding and proposing a model of the construct. Searches for qualitative studies of patients' experiences of COPD-related anxiety were conducted independently by two authors in the databases of PubMed (MEDLINE), CINAHL (EBSCO), and PsycInfo (APA). English-language studies including patients diagnosed with COPD were reviewed, and data were analyzed using thematic analysis. A total of 41 studies were included in the review. Four themes related to COPD-related anxiety were identified: initial events; internal maintaining factors; external maintaining factors; and behavioral maintaining factors. Based on the identified four themes, a conceptual model of COPD-related anxiety from the patient perspective was developed. A conceptual model of COPD-related anxiety from the patient perspective is now available, with the potential to inform future attempts at improving identification and management of COPD-related anxiety. Future research should focus on the development of a COPD-specific anxiety questionnaire containing domains that are relevant from the patient perspective.


Targeting the NLRP3 inflammasome for the treatment of hypertensive target organ damage: Role of natural products and formulations.
Hypertension is a major global health problem that causes target organ damage (TOD) in the heart, brain, kidney, and blood vessels. The mechanisms of hypertensive TOD are not fully understood, and its treatment is challenging. This review provides an overview of the current knowledge on the role of Nod-like receptor pyrin domain containing 3 (NLRP3) inflammasome in hypertensive TOD and the natural products and formulations that inhibit it. We searched PubMed, Web of Science, Google Scholar, and CNKI for relevant articles using the keywords "hypertension," "target organ damage," "NLRP3 inflammasome," "natural products," and "formulations." We reviewed the effects of the NLRP3 inflammasome on hypertensive TOD in different organs and discussed the natural products and formulations that modulate it. In hypertensive TOD, the NLRP3 inflammasome is activated by various stimuli such as oxidative stress and inflammation. Activation of NLRP3 inflammasome leads to the production of pro-inflammatory cytokines that exacerbate tissue damage and dysfunction. Natural products and formulations, including curcumin, resveratrol, triptolide, and allicin, have shown protective effects against hypertensive TOD by inhibiting the NLRP3 inflammasome. The NLRP3 inflammasome is a promising therapeutic target in hypertensive TOD. Natural products and formulations that inhibit the NLRP3 inflammasome may provide novel drug candidates or therapies for hypertensive TOD. Further studies are needed to elucidate the molecular mechanisms and optimize the dosages of these natural products and formulations and evaluate their clinical efficacy and safety.


Social perceptions of cancer and their impacts: implications for nursing practice arising from the literature.
Social perceptions of cancer and their impacts: implications for nursing practice arising from the literature At the millennium cancer still holds a special mystique and is imbued with socio-cultural meanings, which extend far beyond the rational, scientific and biological facts of the disease. Excessive fear and dread may cause family and friends to display avoidance or overprotective behaviours to the ill person, who may subsequently perceive dissatisfaction with social support. Drawing on a literature review this paper explores the impact of cancer on social relationships. Interpersonal strain in relationship is often explained in the stigmatization of the illness and this concept is explored through contemporary social theorizing. These findings have direct implications for nursing practice where the goal of care is to enhance the support relationship.


Daxx maintains endogenous retroviral silencing and restricts cellular plasticity in vivo.
Tumor sequencing studies have emphasized the role of epigenetics and altered chromatin homeostasis in cancer. Mutations in DAXX, which encodes a chaperone for the histone 3.3 variant, occur in 25% of pancreatic neuroendocrine tumors (PanNETs). To advance our understanding of physiological functions of Daxx, we developed a conditional Daxx allele in mice. We demonstrate that Daxx loss is well tolerated in the pancreas but creates a permissive transcriptional state that cooperates with environmental stress (inflammation) and other genetic lesions (Men1 loss) to alter gene expression and cell state, impairing pancreas recovery from inflammatory stress in vivo. The transcriptional changes are associated with dysregulation of endogenous retroviral elements (ERVs), and dysregulation of endogenous genes near ERVs is also observed in human PanNETs with DAXX mutations. Our results reveal a physiologic function of DAXX, provide a mechanism associated with impaired tissue regeneration and tumorigenesis, and expand our understanding of ERV regulation in somatic cells.


Direct observation of ultrafast hydrogen bond strengthening in liquid water.
Water is one of the most important, yet least understood, liquids in nature. Many anomalous properties of liquid water originate from its well-connected hydrogen bond network1, including unusually efficient vibrational energy redistribution and relaxation2. An accurate description of the ultrafast vibrational motion of water molecules is essential for understanding the nature of hydrogen bonds and many solution-phase chemical reactions. Most existing knowledge of vibrational relaxation in water is built upon ultrafast spectroscopy experiments2-7. However, these experiments cannot directly resolve the motion of the atomic positions and require difficult translation of spectral dynamics into hydrogen bond dynamics. Here, we measure the ultrafast structural response to the excitation of the OH stretching vibration in liquid water with femtosecond temporal and atomic spatial resolution using liquid ultrafast electron scattering. We observed a transient hydrogen bond contraction of roughly 0.04 Å on a timescale of 80 femtoseconds, followed by a thermalization on a timescale of approximately 1 picosecond. Molecular dynamics simulations reveal the need to treat the distribution of the shared proton in the hydrogen bond quantum mechanically to capture the structural dynamics on femtosecond timescales. Our experiment and simulations unveil the intermolecular character of the water vibration preceding the relaxation of the OH stretch.


Physician Satisfaction with Anti-Inflammatory Topical Medications for the Treatment of Dry Eye Disease.
Dry eye disease (DED) is one of the most common ophthalmic disorders. Pathogenesis of the disease includes inflammation of the ocular surface and lacrimal gland. Two anti-inflammatory prescription treatments are currently available: cyclosporine ophthalmic emulsion 0.05% (CYC) and lifitegrast 5% ophthalmic solution (LIF). The objective of this survey-based study was to assess physician satisfaction with CYC and LIF for the treatment of DED. Physicians currently treating DED patients with CYC or LIF were asked to rate the experiences of their patients currently or formerly using CYC and LIF, and their own perspectives on the two treatments. Twenty-one physicians participated in the survey, providing responses on behalf of 210 patients. Overall, physicians reported low levels of satisfaction with onset of action of CYC and LIF, and fewer than half considered either drug to be effective in managing symptoms or improving patient quality of life (QoL). Burning sensation and dysgeusia were the most frequently reported side effects. Onset of action and effectiveness after onset were the main switching drivers. Although two-thirds of physicians were satisfied with the overall effectiveness of CYC and LIF, all physicians agreed that more DED treatment options are needed, with >50% strongly agreeing. Physicians perceived a gap in DED management with currently available topical anti-inflammatory agents. Although satisfaction with CYC and LIF was high, few physicians considered these medications to be effective in managing symptoms or improving QoL.


Red blood cells mediated delivery of 9-(2-phosphonylmethoxyethyl)adenine to primary macrophages: efficiency metabolism and activity against human immunodeficiency virus or herpes simplex virus.
Red blood cells (RBC) may act as selective carriers of drugs to macrophages, an important reservoir of viruses such as human immunodeficiency virus (HIV) and herpes simplex virus type 1 (HSV-1). We therefore assessed the incorporation of 9-(2-phosphonylmethoxyethyl)adenine (PMEA), a potent inhibitor of HIV and HSV-1) into RBC, its delivery to macrophages and its activity against HIV or HSV-1. Loading of PMEA in artificially aged opsonized RBC affords significant levels of intracellular PMEA. RBC metabolize PMEA to its active congener PMEA-diphosphate, although with low efficiency. Exposure of macrophages to RBC-encapsulated PMEA inhibits the replication of both HIV and HSV-1 (about 90% inhibition at the highest RBC:macrophages ratios) even if RBC were removed before virus challenge. By contrast, the antiviral activity of free PMEA removed before virus challenge was irrelevant at concentrations up to 150-fold higher than the 50% effective concentration (EC50). Finally, the antiviral effect of RBC-encapsulated PMEA correlates with PMEA levels in macrophages about 500-fold higher than those achieved by free PMEA (at concentrations 10-fold higher than the EC50). The efficacy of RBC-mediated delivery to macrophages of PMEA (and perhaps of compounds with shorter intracellular half-lives) warrants further studies in infectious diseases involving phagocytizing cells as main targets of the pathogen.


Why perinatal mortality cannot be a proxy for maternal mortality.
In recent years, the perinatal mortality rate (PNMR) has been proposed as a proxy measure of maternal mortality, because perinatal deaths are more frequent and potentially more easily measured. This report assesses evidence for an association between these two statistics. This study, based upon data from Matlab, Bangladesh, shows that the maternal mortality ratio (MMR) and the PNMR do not vary together over time, and that the PNMR does not reliably indicate either the magnitude or the direction of change in the MMR from year to year. Statistical analysis shows that the correlation between the PNMR and the MMR is not significantly different from zero. An examination of the major causes of maternal and perinatal deaths indicates that the two measures cannot be expected to vary together. Almost half of perinatal deaths result from causes that do not pose a threat to the mother's life, and almost half of maternal deaths result from causes that do not lead to perinatal death. Monitoring of the PNMR can give an inaccurate picture of maternal mortality and should not be used as a proxy.


The work environment disability-adjusted life year for use with life cycle assessment: a methodological approach.
Life cycle assessment (LCA) is a systems-based method used to determine potential impacts to the environment associated with a product throughout its life cycle. Conclusions from LCA studies can be applied to support decisions regarding product design or public policy, therefore, all relevant inputs (e.g., raw materials, energy) and outputs (e.g., emissions, waste) to the product system should be evaluated to estimate impacts. Currently, work-related impacts are not routinely considered in LCA. The objectives of this paper are: 1) introduce the work environment disability-adjusted life year (WE-DALY), one portion of a characterization factor used to express the magnitude of impacts to human health attributable to work-related exposures to workplace hazards; 2) outline the methods for calculating the WE-DALY; 3) demonstrate the calculation; and 4) highlight strengths and weaknesses of the methodological approach. The concept of the WE-DALY and the methodological approach to its calculation is grounded in the World Health Organization's disability-adjusted life year (DALY). Like the DALY, the WE-DALY equation considers the years of life lost due to premature mortality and the years of life lived with disability outcomes to estimate the total number of years of healthy life lost in a population. The equation requires input in the form of the number of fatal and nonfatal injuries and illnesses that occur in the industries relevant to the product system evaluated in the LCA study, the age of the worker at the time of the fatal or nonfatal injury or illness, the severity of the injury or illness, and the duration of time lived with the outcomes of the injury or illness. The methodological approach for the WE-DALY requires data from various sources, multi-step instructions to determine each variable used in the WE-DALY equation, and assumptions based on professional opinion. Results support the use of the WE-DALY in a characterization factor in LCA. Integrating occupational health into LCA studies will provide opportunities to prevent shifting of impacts between the work environment and the environment external to the workplace and co-optimize human health, to include worker health, and environmental health.


3D bioprinting for engineering complex tissues.
Bioprinting is a 3D fabrication technology used to precisely dispense cell-laden biomaterials for the construction of complex 3D functional living tissues or artificial organs. While still in its early stages, bioprinting strategies have demonstrated their potential use in regenerative medicine to generate a variety of transplantable tissues, including skin, cartilage, and bone. However, current bioprinting approaches still have technical challenges in terms of high-resolution cell deposition, controlled cell distributions, vascularization, and innervation within complex 3D tissues. While no one-size-fits-all approach to bioprinting has emerged, it remains an on-demand, versatile fabrication technique that may address the growing organ shortage as well as provide a high-throughput method for cell patterning at the micrometer scale for broad biomedical engineering applications. In this review, we introduce the basic principles, materials, integration strategies and applications of bioprinting. We also discuss the recent developments, current challenges and future prospects of 3D bioprinting for engineering complex tissues. Combined with recent advances in human pluripotent stem cell technologies, 3D-bioprinted tissue models could serve as an enabling platform for high-throughput predictive drug screening and more effective regenerative therapies.


Differences in muscle transcriptome among pigs phenotypically extreme for fatty acid composition.
Besides having an impact on human health, the porcine muscle fatty acid profile determines meat quality and taste. The RNA-Seq technologies allowed us to explore the pig muscle transcriptome with an unprecedented detail. The aim of this study was to identify differentially-expressed genes between two groups of 6 sows belonging to an Iberian × Landrace backcross with extreme phenotypes according to FA profile. We sequenced the muscle transcriptome acquiring 787.5 M of 75 bp paired-end reads. About 85.1% of reads were mapped to the reference genome. Of the total reads, 79.1% were located in exons, 6.0% in introns and 14.9% in intergenic regions, indicating expressed regions not annotated in the reference genome. We identified a 34.5% of the intergenic regions as interspersed repetitive regions. We predicted a total of 2,372 putative proteins. Pathway analysis with 131 differentially-expressed genes revealed that the most statistically-significant metabolic pathways were related with lipid metabolism. Moreover, 18 of the differentially-expressed genes were located in genomic regions associated with IMF composition in an independent GWAS study in the same genetic background. Thus, our results indicate that the lipid metabolism of FAs is differently modulated when the FA composition in muscle differs. For instance, a high content of PUFA may reduce FA and glucose uptake resulting in an inhibition of the lipogenesis. These results are consistent with previous studies of our group analysing the liver and the adipose tissue transcriptomes providing a view of each of the main organs involved in lipid metabolism. The results obtained in the muscle transcriptome analysis increase the knowledge of the gene regulation of IMF deposition, FA profile and meat quality, in terms of taste and nutritional value. Besides, our results may be important in terms of human health.


Transition from a heterozygous to a homozygous state of a pair of loci in the inverted repeat sequences of the L component of the herpes simplex virus type 1 genome.
The behavior of herpes simplex virus type 1 heterozygous isolates, in which the two inverted repeats of the L component (RL) were differentiated by a polymorphism marker (the presence [type B] or absence [type A] of a SalI site), was investigated. The progeny viruses derived from the heterozygote (A/B) consisted of heterozygotes (A/B), type A homozygotes (A/A), and type B homozygotes (B/B). The heterology between RL, albeit tolerated, was unstable, as is the case with heterology between the repeats of the S component. The two repeats TRL (terminal) and IRL (internal) were equipotent in generating homozygotes from a heterozygote. Data obtained from an analysis of 426 progeny viruses derived from heterozygous clones supported the hypothesis that the two loci in RL of a herpes simplex virus type 1 genome are determined as a random combination of the corresponding two loci in RL of the parent virus and that the ratio of heterozygotes/type A homozygotes/type B homozygotes in the progeny viruses from a heterozygote is expected to be 2:1:1. An ephemeral dominance of one type of homozygote over the other was observed in subclones from several heterozygous clones.


The Society of Thoracic Surgeons Congenital Heart Surgery Database: 2016 Update on Outcomes and Quality.
The Society of Thoracic Surgeons Congenital Heart Surgery Database is the largest congenital and pediatric cardiac surgical clinical data registry in the world. It is the platform for all activities of The Society of Thoracic Surgeons related to the analysis of outcomes and the improvement of quality in this subspecialty. This article summarizes current aggregate national outcomes in congenital and pediatric cardiac surgery and reviews related activities in the areas of quality measurement, performance improvement, and transparency. The reported data about aggregate national outcomes are exemplified by an analysis of 10 benchmark operations performed from January 2011 to December 2014 and documenting overall discharge mortality (interquartile range among programs with more than 9 cases): off-bypass coarctation, 1.0% (0.0% to 0.9%); ventricular septal defect repair, 0.7% (0.0% to 1.1%); tetralogy of Fallot repair, 1.0% (0.0% to 1.7%); complete atrioventricular canal repair, 3.2% (0.0% to 6.5%); arterial switch operation, 2.7% (0.0% to 5.6%); arterial switch operation plus ventricular septal defect, 5.3% (0.0% to 6.7%); Glenn/hemiFontan, 2.1% (0.0% to 3.8%); Fontan operation, 1.4% (0.0% to 2.4%); truncus arteriosus repair, 9.6% (0.0 % to 11.8%); and Norwood procedure, 15.6% (10.0% to 21.4%).


Ubiquitin-proteasome system stress sensitizes ovarian cancer to proteasome inhibitor-induced apoptosis.
The ubiquitin-proteasome system (UPS) mediates targeted protein degradation. Notably, the UPS determines levels of key checkpoint proteins controlling apoptosis and proliferation by controlling protein half-life. Herein, we show that ovarian carcinoma manifests an overstressed UPS by comparison with normal tissues by accumulation of ubiquitinated proteins despite elevated proteasome levels. Elevated levels of total ubiquitinated proteins and 19S and 20S proteasome subunits are evident in both low-grade and high-grade ovarian carcinoma tissues relative to benign ovarian tumors and in ovarian carcinoma cell lines relative to immortalized surface epithelium. We find that ovarian carcinoma cell lines exhibit greater sensitivity to apoptosis in response to proteasome inhibitors than immortalized ovarian surface epithelial cells. This sensitivity correlates with increased cellular proliferation rate and UPS stress rather than absolute proteasome levels. Proteasomal inhibition in vitro induces cell cycle arrest and the accumulation of p21 and p27 and triggers apoptosis via activation of caspase-3. Furthermore, treatment with the licensed proteasome inhibitor PS-341 slows the growth of ES-2 ovarian carcinoma xenograft in immunodeficient mice. In sum, elevated proliferation and metabolic rate resulting from malignant transformation of the epithelium stresses the UPS and renders ovarian carcinoma more sensitive to apoptosis in response to proteasomal inhibition.


Design of a study to determine the impact of insecticide resistance on malaria vector control: a multi-country investigation.
Progress in reducing the malaria disease burden through the substantial scale up of insecticide-based vector control in recent years could be reversed by the widespread emergence of insecticide resistance. The impact of insecticide resistance on the protective effectiveness of insecticide-treated nets (ITN) and indoor residual spraying (IRS) is not known. A multi-country study was undertaken in Sudan, Kenya, India, Cameroon and Benin to quantify the potential loss of epidemiological effectiveness of ITNs and IRS due to decreased susceptibility of malaria vectors to insecticides. The design of the study is described in this paper. Malaria disease incidence rates by active case detection in cohorts of children, and indicators of insecticide resistance in local vectors were monitored in each of approximately 300 separate locations (clusters) with high coverage of malaria vector control over multiple malaria seasons. Phenotypic and genotypic resistance was assessed annually. In two countries, Sudan and India, clusters were randomly assigned to receive universal coverage of ITNs only, or universal coverage of ITNs combined with high coverage of IRS. Association between malaria incidence and insecticide resistance, and protective effectiveness of vector control methods and insecticide resistance were estimated, respectively. Cohorts have been set up in all five countries, and phenotypic resistance data have been collected in all clusters. In Sudan, Kenya, Cameroon and Benin data collection is due to be completed in 2015. In India data collection will be completed in 2016. The paper discusses challenges faced in the design and execution of the study, the analysis plan, the strengths and weaknesses, and the possible alternatives to the chosen study design.


Negative regulation of FcepsilonRI signaling by FcgammaRII costimulation in human blood basophils.
Signaling through the antigen receptors of human B and T cells and the high-affinity IgE receptor FcepsilonRI of rodent mast cells is decreased by cross-linking these receptors to the low-affinity IgG receptor FcgammaRII. The inhibition is thought to involve the tyrosine phosphorylation of immunoreceptor tyrosine-based inhibitory motifs (ITIMs) in the FcgammaRIIB cytoplasmic tail, creating binding sites for SH2-containing protein (Src homology domain containing protein tyrosine phosphatase 1 and 2 [SHP-1, SHP-2]) and/or lipid (SH2 domain-containing polyphosphatidyl-inositol 5-phosphatase) phosphatases that oppose activating signals from the costimulated antigen receptors. In human basophils and mast cells FcepsilonRI signaling generates mediators and cytokines responsible for allergic inflammation. We proposed to determine whether FcepsilonRI signaling is inhibited by FcgammaRII costimulation in human basophils and to explore the underlying mechanism as an approach to improving the treatment of allergic inflammation. FcgammaR expression on human basophils was examined using flow cytometry and RT-PCR analysis. FcgammaRII/FcepsilonRI costimulation was typically accomplished by priming cells with anti-dinitrophenol (DNP) IgE and anti-DNP IgG and stimulating with DNP-BSA. Phosphatases were identified by Western blotting, and their partitioning between membrane and cytosol was determined by cell fractionation. Biotinylated synthetic peptides and phosphopeptides corresponding to the FcgammaRIIB ITIM sequence were used for adsorption assays. We report that peripheral blood basophils express FcgammaRII (in both the ITIM-containing FcgammaRIIB and the immunoreceptor tyrosine-based activation motif-containing FcgammaRIIA forms) and that costimulating FcgammaRII and FcepsilonRI inhibits basophil FcepsilonRI-mediated histamine release, IL-4 production, and Ca(2+) mobilization. The inhibition of basophil FcepsilonRI signaling by FcgammaRII/FcepsilonRI costimulation is linked to a significant decrease in Syk tyrosine phosphorylation. Human basophils express all 3 SH2-containing phosphatases. Evidence that FcgammaRII/FcepsilonRI costimulation induces SHP-1 translocation from the cytosolic to membrane fractions of basophils and that biotinylated synthetic peptides corresponding to the phosphorylated FcgammaRIIB ITIM sequence specifically recruit SHP-1 from basophil lysates particularly implicates this protein phosphatase in the negative regulation of FcepsilonRI signaling by costimulated FcgammaRII.


Toxic encephalopathy.
Neurotoxic exposures are common. Although there are myriad substances that can cause encephalopathy, this review focuses on common environmental neurotoxins, such as select heavy metals, organic industrial toxins, and pesticides. The central nervous system is susceptible to toxic injury, and many environmental neurotoxins are capable of causing encephalopathy. When a patient presents with toxic encephalopathy, the differential diagnosis is initially broad. The clinical presentation after exposure to a toxin varies in severity among patients. Arriving at the correct diagnosis is often a diagnostic challenge. The importance of taking a good history and performing a comprehensive examination cannot be overemphasized. Neuroimaging and neurophysiologic testing typically play ancillary roles. Confirmatory laboratory testing is available for some toxins. Treatment of most toxic encephalopathies is not supported by clinical trials; additional research is needed in the field.


Effect of pressurized hot water extraction on antioxidants from grape pomace before and after enological fermentation.
Grape pomace was extracted with pressurized hot water at laboratory scale before and after fermentation to explore the effects of fermentation and extraction temperature (50-200 °C) and time (5 and 30 min) on total extracted antioxidant levels and activity and to determine the content and recovery efficiency of main grape polyphenols, anthocyanins, and tannins. Fermented pomace yielded more total antioxidants (TAs), antioxidant activity, and tannins, than unfermented pomace but fewer anthocyanins. Elevating the extraction temperature increased TA extraction and antioxidant activity. Maximum anthocyanin extraction yields were achieved at 100 °C and at 150 °C for tannins and tannin-anthocyanin adducts. Using higher temperatures and longer extraction times resulted in a sharp decrease of polyphenol extraction yield. Relevant proanthocyanidin amounts were extracted only at 50 and 100 °C. Finally, TA recovery and activity were not directly related to the main polyphenol content when performing pressurized hot water grape pomace extraction.


Compulsive hoarding in children.
This article discusses the nature and treatment of compulsive hoarding among children. We summarize the phenomenology of compulsive hoarding, including its clinical presentation, comorbidity with varied mental disorders, and associated impairment. The limited data on treatment outcome are presented along with a behavioral framework that we utilized to treat youth who hoard. Our approach is highlighted in the context of a case illustration of an 11-year-old girl suffering from compulsive hoarding and several comorbid mental health disorders. We conclude with recommendations for clinical work with this challenging and neglected population.


Prognostic value of estrogen receptor status can be improved by combined evaluation of p53, Bcl2 and PgR expression: an immunohistochemical study on breast carcinoma with long-term follow-up.
Steroid receptor analysis is the only widely accepted prognostic/predictive marker in breast cancer (BC) treatment. In the present study we evaluated the prognostic role of ER/PgR with p53 and Bcl2, in a series of 277 BC (153 pN1/2, 122 pNO, 2 pNx) with a long-term follow-up (67 months for DFS, 75 months for OS). Our results, besides confirming the usefulness of ER immunohistochemical expression as a prognostic marker, showed that PgR expression alone had a borderline/not significant prognostic value in the whole series (p=0.08 for DFS and p=0.09 for OS), while showed to be prognostic in N+ cases (p=0.02 for DFS and p=0.03 for OS). PgR prognostic value, however, was not independent at the multivariate analysis. By combining ER with PgR, p53 and Bcl2, we showed that ER/p53 and ER/Bcl2 phenotypes had a better discriminant role than ER/PgR phenotype. The ER+/p53+ phenotype was at higher risk of relapse/death as compared with ER+/p53- phenotype. Conversely ER-/p53+ phenotype showed the most unfavourable prognosis. Similar results could be observed concerning ER/Bcl2 phenotypes. Our study showed that the combined evaluation of ER/PgR weakly enhanced the prognostic/predictive power of ER status alone. On the contrary, the combined evaluation of ER/p53 and ER/Bcl2, improved this prognostic/predictive capability and allowed the separation of ER positive and ER negative cases into subgroups with different prognosis.


Teen Dating Violence Victimization: Associations Among Peer Justification, Attitudes Toward Gender Inequality, Sexual Activity, and Peer Victimization.
The current study, grounded in a social learning theoretical framework, examined attitudes and behaviors associated with verbal and physical teen dating violence (TDV) victimization. Because TDV varies by gender in both frequency and severity, these associations were examined first within the overall sample, and then by gender to further investigate these differences. A total of 1,884 adolescents (49.2% boys; 50.8% girls; average age 14.79 years; SD = .58) who reported ever dating someone were included in the analysis. Specifically, peers' justification of TDV, attitudes supporting gender inequality, sexual activity, and peer victimization were included to determine their cross-sectional association with verbal and physical TDV victimization. Data were analyzed separately for boys and girls. Results indicated that peers' justification of TDV, peer victimization, sexual activity, and attitudes supporting gender inequality were each associated with higher physical and verbal TDV victimization for girls and boys. Most of these factors remained significant when separated by gender, except for sexual activity and attitudes supporting gender inequality, which were not associated with physical TDV victimization for boys and girls, respectively. Implications for practice and research are discussed.


Effect of fatty acid oxidation on efficiency of energy production in rat heart.
Myocardial fatty acid oxidation has been reported to be accompanied by an elevated O2 consumption compared with carbohydrate oxidation. The exact amount of this additional O2 consumption is controversial. Different investigators have observed an O2 wasting effect that is too large to be explained by the different ATP-to-O2 ratios of these substrates. With the use of isolated perfused rat hearts, O2 consumption and hemodynamic measurements were computer analyzed to provide on-line estimates of the ratio between O2 consumption and demand (EQ). Increasing palmitate or octanoate concentrations decreased the respiratory quotient, which was accompanied by a disproportionate increase of EQ. Inhibition of fatty acid oxidation by an inhibitor of acylcarnitine transferase or a blockade of mitochondrial thiolase caused a drastic reduction of fatty acid oxidation. The fatty acid-induced enhancement of O2 consumption was decreased to a much smaller extent, indicating that there are two different mechanisms responsible for the O2-wasting effect, one that depends on mitochondrial fatty acid oxidation and another that is not affected by an inhibition of this pathway.


Beef Fat Enriched with Polyunsaturated Fatty Acid Biohydrogenation Products Improves Insulin Sensitivity Without Altering Dyslipidemia in Insulin Resistant JCR:LA-cp Rats.
The main dietary sources of trans fatty acids are partially hydrogenated vegetable oils (PHVO), and products derived from polyunsaturated fatty acid biohydrogenation (PUFA-BHP) in ruminants. Trans fatty acid intake has historically been associated with negative effects on health, generating an anti-trans fat campaign to reduce their consumption. The profiles and effects on health of PHVO and PUFA-BHP can, however, be quite different. Dairy products naturally enriched with vaccenic and rumenic acids have many purported health benefits, but the putative benefits of beef fat naturally enriched with PUFA-BHP have not been investigated. The objective of the present experiment was to determine the effects of beef peri-renal fat (PRF) with differing enrichments of PUFA-BHP on lipid and insulin metabolism in a rodent model of dyslipidemia and insulin resistance (JCR:LA-cp rat). The results showed that 6 weeks of diet supplementation with beef PRF naturally enriched due to flaxseed (FS-PRF) or sunflower-seed (SS-PRF) feeding to cattle significantly improved plasma fasting insulin levels and insulin sensitivity, postprandial insulin levels (only in the FS-PRF) without altering dyslipidemia. Moreover, FS-PRF but not SS-PRF attenuated adipose tissue accumulation. Therefore, enhancing levels of PUFA-BHP in beef PRF with FS feeding may be a useful approach to maximize the health-conferring value of beef-derived fats.


An evaluation of antisocial behaviour in children after traumatic brain injury: the prospect of improving the quality of life in rehabilitation.
The aim of the article is to present the consequences of traumatic brain injury in children, associated with general cognition and behavioural disorders, mainly of the antisocial type. A total of 20 school-age children took part in the study, including six girls and 14 boys. The average age of the children was 13.35 years (standard deviation SD = 1.95). The research instruments included an analysis of documentation, a structured clinical interview, MMSE and Frontal Behavioral Inventory (FBInv) with additional set of five supplementary questions directed for detection of antisocial behavior. The research was conducted from the beginning of January 2009 until the end of May 2009. As hypothesized, the functioning of the children with traumatic brain injury is severely disrupted, because of the presence of cognitive impairment, however, dementia is not manifested. In a significant number of the children with traumatic brain injury we found not only the frontal syndrome, but also the occurrence of antisocial behaviour. The most commonly reported behavioural problems were: disorganization commonly referred to as laziness, hypersensitivity, and anxiety. The most common types of anti-social behaviour were: impulsivity, physical and verbal aggression, and also an outburst of anger. The children with traumatic brain injury suffer from a cognitive disorders and behavioural problems, especially impulsivity, physical and verbal aggression, increased anxiety, and disorganization. The occurrence of frontal syndrome is related to the development of antisocial behaviour.


Clinicopathological significance of caspase-8 and caspase-10 expression in rectal cancer.
To investigate the expression of caspase-8 and -10 in rectal adenoma, adenocarcinoma and the corresponding normal mucosa tissue, and to clarify the relationship between their expression and clinicopathological parameters of rectal cancer. The expression of caspase-8 and -10 was determined by real-time RT-PCR and immunohistochemistry in 36 rectal adenomas, 93 rectal cancers and 93 corresponding normal rectal mucosa samples. Compared with normal mucosa, the mRNA expression of caspase-8 was higher in adenomas (p = 0.003), while that of caspase-10 was lower in adenomas (p = 0.035) and cancers (p = 0.001). Immunohistochemical results showed caspase-8 up-regulation in adenomas (p = 0.014), and caspase-10 down-regulation in adenomas (p = 0.034) and cancers (p < 0.001) compared with normal mucosa samples. Cancers with poor differentiation had lower caspase-10 mRNA and protein levels than those with better differentiation (p = 0.041 and p = 0.046, respectively). The protein expression of caspase-8 and -10 was in accordance with the mRNA expression (p = 0.043 and p = 0.018, respectively). Caspase-8 expression was up-regulated in rectal adenomas. Caspase-10 expression was down-regulated in both rectal adenomas and cancers, and was further related to differentiation. Caspase-8 and -10 may be involved in the pathogenesis of rectal cancer.


Gene expression analysis reveals novel altered genes in nasal polyps.
Nasal polyposis (NP) is a chronic inflammatory disease of the nasal mucosa characterized by the infiltration of inflammatory cells, mainly eosinophils. Although nasal polyposis occurs in 4% of the population, its physiopathology remains unclear. The aim of this study was to identify and characterize differentially expressed genes that can be used in the prognosis, treatment and elucidation of this physiopathology. To identify novel genes differentially expressed in NP, we applied real-time quantitative PCR to 11 NP samples and to a pool of total RNA from a subset of 13 normal nasal mucosa samples from human autopsies. For selecting genes, the methylated CpG island amplification technique was used. Five differentially methylated clones (ATP2A1, NOVA1, PLCD3, SOLH and TGFβI) were identified. However, these genes presented methylated CpG islands between exons, i.e., not in the promoter regions of the genes. Thus, as shown by real-time PCR, the ATP2A1, SOLH, PLDC3 and TGFβI genes were overexpressed in NP. The genes identified in this study are probably involved in some stage of the process of formation and development of nasal polyposis, as they were highly expressed in the nasal polyp samples.


Endogenous Calcitriol Synthesis Controls the Humoral IgE Response in Mice.
The vitamin D receptor participates in the control of IgE class-switch recombination in B cells. The physiologic vitamin D receptor agonist, 1,25(OH)2D3 (calcitriol), is synthesized by the essential enzyme 25-hydroxyvitamin D3-1α-hydroxylase (CYP27B1), which can be expressed by activated immune cells. The role of endogenous calcitriol synthesis for the regulation of IgE has not been proven. In this study, we investigated IgE-responses in Cyp27b1-knockout (KO) mice following sensitization to OVA or intestinal infection with Heligmosomoides polygyrus Specific Igs and plasmablasts were determined by ELISA and ELISpot, Cyp27b1 expression was measured by quantitative PCR. The data show elevated specific IgE and IgG1 concentrations in the blood of OVA-sensitized Cyp27b1-KO mice compared with wild-type littermates (+898 and +219%). Accordingly, more OVA-specific IgG1-secreting cells are present in spleen and fewer in the bone marrow of Cyp27b1-KO mice. Ag-specific mechanisms are suggested as the leucopoiesis is in general unchanged and activated murine B and T lymphocytes express Cyp27b1 Accordingly, elevated specific IgE concentrations in the blood of sensitized T cell-specific Cyp27b1-KO mice support a lymphocyte-driven mechanism. In an independent IgE-inducing model, i.e., intestinal infection with H. polygyrus, we validated the increase of total and specific IgE concentrations of Cyp27b1-KO compared with wild-type mice, but not those of IgG1 or IgA. We conclude that endogenous calcitriol has an impact on the regulation of IgE in vivo. Our data provide genetic evidence supporting previous preclinical and clinical findings and suggest that vitamin D deficiency not only promotes bone diseases but also type I sensitization.


A PAT-based qualification of pharmaceutical excipients produced by batch or continuous processing.
Pharmaceutical excipients have an influence on the main requirements for medicinal products (viz., quality, safety and efficacy) but also on their manufacturability. During product lifecycle it may become necessary to introduce minor changes (e.g., to continuously improve it) or major changes in the validated process (e.g., moving it to a new production site, replacing process version or even disruptively changing processing type). Those changes can influence the critical to quality attributes of the product. Therefore, it is important to enhance process understanding to avoid the risk of any significant quality changes. Process analytical technology can support better decision making and risk-management as required in quality by design - viz., by many pharmaceutical regulatory authorities. This study compares the quality of the pharmaceutical excipient sodium carbonate (anhydrous) produced either in a batch or a continuous process. For continuous processing two different production lines were available that differed on the dryer and crystallizer types used. Therefore their influence on critical to quality attributes of sodium carbonate was investigated for each of the three processing alternatives. The overall goal was to identify which of the continuous processes ensures a similar product quality to batch processing. Namely, changes on chemical and physical attributes of the product were investigated with Raman spectroscopy, laser diffraction and X-ray powder diffraction. Principal component analysis, a very common multivariate analysis technique, was applied to extract relevant information from small differences at multiple spectral regions from samples from each process type and from each analytical technique used. Changing processing from batch to continuous improved consistency of certain attributes (e.g., particle size distribution) but affected others. However, the increased process/product knowledge gained can lead to an enhanced control strategy and ensure a similar product quality is obtained from distinct process versions.


Recent rodent models for Alzheimer's disease: clinical implications and basic research.
Alzheimer's disease (AD) is the most common origin of dementia in the elderly. Although the cause of AD remains unknown, several factors have been identified that appear to play a critical role in the development of this debilitating disorder. In particular, amyloid precursor protein (APP), tau hyperphosphorylation, and the secretase enzymes, have become the focal point of recent research. Over the last two decades, several transgenic and non-transgenic animal models have been developed to elucidate the mechanistic aspects of AD and to validate potential therapeutic targets. Transgenic rodent models over-expressing human β-amyloid precursor protein (β-APP) and mutant forms of tau have become precious tools to study and understand the pathogenesis of AD at the molecular, cellular and behavioural levels, and to test new therapeutic agents. Nevertheless, none of the transgenic models of AD recapitulate fully all of the pathological features of the disease. Octodon degu, a South American rodent has been recently found to spontaneously develop neuropathological signs of AD in old age. This review aims to address the limitations and clinical relevance of transgenic rodent models in AD, and to highlight the potential for O. degu as a natural model for the study of AD neuropathology.