Serpulina hyodysenteriae challenge of fattening pigs vaccinated with anadjuvanted bivalent bacterin against swine dysentery.
An experimental fattening period was performed with 36 pigs in order to evaluate the safety and efficacy of a bivalent adjuvanted Serpulina hyodysenteriae bacterin. The pigs originated from three different farms and were split into six groups. Eighteen pigs were vaccinated twice, two weeks apart and the rest served as controls. One unvaccinated pig in each group was inoculated intragastrically with 4 x 10(8) active S. hyodysenteriae spirochaetes, B 78 strain. Weight evolution, clinical symptoms, shedding of S. hyodysenteriae and serum antibody titres were monitored for 12 weeks in all pigs. The severity of clinical symptoms and shedding of spirochaetes in faeces were significantly reduced in vaccinated pigs as compared to controls. Statistically significant differences in weight gain between vaccinates and controls were detected coinciding with the maximum shedding of S. hyodysenteriae in faeces. All vaccinated pigs seroconverted, as measured by indirect ELISA, and had a good anamnestic response after the second vaccine dose. The bacterin proved to be useful for both the reduction of clinical symptoms and the number of infected animals in a fattening unit. Adverse post-vaccinal reactions were not observed in any pig.


Analysis of local extracellular matrix identifies different aetiologies behind bicuspid and tricuspid aortic valve degeneration and suggests therapies.
Aortic valve degeneration (AVD) is a life-threatening condition that has no medical treatment and lacks individual therapies. Although extensively studied with standard approaches, aetiologies behind AVD are unclear. We compared abundances of extracellular matrix (ECM) proteins from excised valve tissues of 88 patients with isolated AVD of normal tricuspid (TAV) and congenital bicuspid aortic valves (BAV), quantified more than 1400 proteins per ECM sample by mass spectrometry, and demonstrated that local ECM preserves molecular cues of the pathophysiological processes. The BAV ECM showed enrichment with fibrosis markers, namely Tenascin C, Osteoprotegerin, and Thrombospondin-2. The abnormal physical stress on BAV may cause a mechanical injury leading to a continuous Tenascin C-driven presence of myofibroblasts and persistent fibrosis. The TAV ECM exhibited enrichment with Annexin A3 (p = 1.1 × 10-16 and the fold change 6.5) and a significant deficit in proteins involved in high-density lipid metabolism. These results were validated by orthogonal methods. The difference in the ECM landscape suggests distinct aetiologies between AVD of BAV and TAV; warrants different treatments of the patients with BAV and TAV; elucidates the molecular basis of AVD; and implies possible new therapeutic approaches. Our publicly available database (human_avd_ecm.surgsci.uu.se) is a rich source for medical doctors and researchers who are interested in AVD or heart ECM in general. Systematic proteomic analysis of local ECM using the methods described here may facilitate future studies of various tissues and organs in development and disease.


A modified Western blot protocol for enhanced sensitivity in the detection of a membrane protein.
Membrane proteins, owing to their highly hydrophobic nature, have always posed a daunting challenge to biochemists and structural biologists working on the characterization of these "naughty" proteins. Here we describe a problem that we encountered in the immunodetection of a hemagglutinin (HA) epitope-tagged membrane protein, Hgt1p (high-affinity glutathione transporter from the yeast Saccharomyces cerevisiae), for which little or no signal was observed on the blots with monoclonal antibody, on following the standard Western blot protocol. The introduction of a single step that involved posttransfer incubation of the blots with sodium dodecyl sulfate (SDS)/beta-mercaptoethanol solution at 55 degrees C for 15 min enabled us to detect a strong, stable, and reproducible signal for the membrane protein.


Herd immunity in older adults from a middle-income country: A time-series trend analysis of community-acquired pneumonia mortality 2003-2017.
Community-acquired pneumonia is responsible for substantial mortality, and pneumococcus is commonly accepted as a major cause of pneumonia, regardless of laboratory confirmation. Child immunization programs have reported success in decreasing pneumonia mortality: directly in young children and indirectly (herd immunity) in unvaccinated adult populations in some countries. We assess changes in mortality trends for all-cause pneumonia in older adults associated with the introduction of pneumococcal vaccination for children in Peru. This is a secondary analysis on administrative data collected periodically by the Peruvian Ministry of Health. An observational retrospective time series analysis was conducted using longitudinal population-based data from death certificates in Peru between 2003 and 2017. The time series includes 6 years before and 9 years after the introduction of the pneumococcal-conjugated vaccines in the national child immunization program in 2009. Monthly frequencies and annual rates for all-cause pneumonia deaths in children under 5 years of age and adults over 65 years of age are presented. Linear and quadratic trends are analyzed. Deaths among older adults accounted for 75.6% of all-cause pneumonia mortality in Peru, with 94.4% of these reporting "pneumonia due to unspecified organism" as the underlying cause of death. Comparing pre- and post-child immunization program periods, annual average mortality rates from unspecified pneumonia decreased by 22.7% in young children but increased by 19.6% in older adults. A linear trend model supports this overall tendency, but a quadratic curve explains the data better. Pneumococcal-conjugated vaccines are developed using serotypes prevalent in selected countries from less common (invasive) pneumococcal disease and expected to prevent mortality worldwide from widespread (noninvasive) pneumonia. Our results do not support the presence of herd immunity from pneumococcal vaccination of children for community-acquired pneumonia in the increasingly ageing population of Peru. This should direct future research and could influence public health policy.


The role of the nurse in caring for the critical patient with sepsis.
to know the nursing interventions in the identification, prevention and control of sepsis in critical patients. integrative review of the literature, with two parallel researches using different MesH terms, using the EBSCO database and Google Scholar. Nine studies were included in the sample. nursing interventions are centered on the creation/implementation of protocols for the early recognition of sepsis, the training of teams to ensure a safe and effective approach and the adoption of measures for infection prevention and control as a way to prevent sepsis. the evidence shows that nurses are fundamental in the early identification, control and prevention of sepsis, preventing disease progression and contributing to decreased morbidity and mortality.


L-carnosine as an enhancer to computerized cognitive behaviour therapy in Japanese workers, an unjustified claim.
This letter comments on the conclusion drawn by Shirotsuki et al. (2017) in their article entitled "The effect for Japanese workers of a self-help computerized cognitive behaviour therapy program with a supplement soft drink", recently published in BioPsychoSocial Medicine. The authors concluded that their drink, containing L-carnosine, enhances the effects of a computerized cognitive-behavioural therapy (CCBT) on the psychological well-being of healthy Japanese workers. Yet, we argue that their conclusion is unfounded given their results and the methodological shortcomings of their study. Briefly, while the authors reported improvement on the tension-anxiety subscale of the Profile of Mood States (POMS) in the CCBT only group, they also observed a lack of improvement on this subscale in the CCBT+L-carnosine group suggesting that the drink washes out this beneficial effect of CCBT. Methodological issues include the uncontrolled levels of L-carnosine metabolized by participants jeopardize the study's internal validity. Also, the clinical meaningfulness of the findings seems dubious as post-treatment scores remained within the range of the general Japanese population. Consequently, we argue that Shirotsuki et al.'s study should be re-conducted before drawing any valid conclusion.


Effects of inotropic and chronotropic stimuli on acute myocardial ischemic injury. II. Studies with dopamine and ouabain in the barbiturate-anesthetized dog.
To test the hypothesis that selective increases in inotropic state without concomitant acceleration of heart rate would not augment acute ischemic injury in the non-failing heart of the anesthetized dog, we carried out studies in 16 dogs subjected to serial 10-min occlusions of the left anterior descending coronary artery. The severity of ischemic injury was determined by mass spectrometric measurement of the rise in intramural carbon dioxide tension (delta PmCO2) in the ischemic zone, and inotropic stimulation was provided by either dopamine or ouabain. In Group I dogs (n = 9), dopamine [4 +/- 1 (SD) micrograms/kg/min] was infused before the final occlusion to increase left ventricular (LV) dP/dt without changing heart rate; delta PmCO2 was not significantly different between control (64 +/- 21 mm Hg) and postdopamine (67 +/- 22 mm Hg) occlusions. In Group II dogs (n = 7), ouabain (0.03 mg/kg) was administered 15 min before the final occlusion, resulting in a significant increase in LV dP/dt and a slight decrease in heart rate (average 13 beats/min); delta PmCO2 was slightly decreased in the occlusion after ouabain (60 +/- 12 mm Hg) compared with the preceding occlusion without inotropic stimulation (67 +/- 13 mm Hg), p less than 0.05. Throughout the studies in both groups, there were no significant changes in collateral blood flow to the central ischemic zone, or in heart rate-systolic arterial pressure product, an estimate of myocardial oxygen consumption. Analyses of individual responses revealed that when LV dP/dt increased by 50% or more after dopamine or ouabain, ischemia was more likely to intensify.(ABSTRACT TRUNCATED AT 250 WORDS)


Smoking Status Affects the Association Between Hematoma Heterogeneity and Hematoma Expansion.
The purpose of this study was to verify the relationship between hematoma heterogeneity and hematoma expansion and explore any effect modifiers through subgroup analyses. Clinical records of 357 patients with spontaneous cerebral hemorrhage at Shenzhen Second People's Hospital from March 2016 to October 2018 were included in the study. Hematoma heterogeneity was measured on the first noncontrast computed tomography image according to the Barras scale. Hematoma expansion was defined as an absolute hematoma volume increase of 6 mL, or a 33% increase. We performed univariate and multivariate logistic regression analyses, as well as subgroup analyses, to assess the relationship between the presence of heterogeneity on noncontrast computed tomography and hematoma expansion. Hematoma expansion occurred in 79 (22.13%) of the 357 patients with intracerebral hemorrhage (ICH). Among the patients with ICH, there were 83 smokers, accounting for 23.24%. The average patient age was 56.21 ± 13.75 years, and 74.51% were male. Compared with the absence of heterogeneity, the risk of hematoma expansion increased by 1.06 times (odds ratio, 2.06; 95% confidence interval, 1.10-3.86). Based on the subgroup analysis, smoking status was found to modify the association between heterogeneity and hematoma expansion; the association was stronger in smokers than in nonsmokers (odds ratio, 10.23; 95% confidence interval, 2.15-48.65). Heterogeneity independently predicts hematoma expansion, especially in smoking patients.


Depressive states among adults with diabetes: Findings from the National Health and Nutrition Examination Survey, 2007-2012.
To determine (1) the prevalence of SubD states among adults with diabetes, and (2) whether evidence exists of an independent association between diabetes status and SubD, controlling for selected confounders. Data from the 2007-2012 National Health and Nutrition Examination Surveys were combined to estimates of depressive states by diabetes status among the noninstitutionalized U.S. adult population, and to assess the association of diabetes status and depressive states using a polytomous logistic regression model. An estimated 17%, or 3.7 million, of U.S. adults with diabetes (diagnosed and undiagnosed) met criteria for either mD or ssD. The majority of SubD cases with diabetes were found to be ssD (10.1%) compared with mD (6.9%). After controlling for the effects of age, sex, race and ethnicity, education, body mass index, and poverty as covariates, an independent association persists between diagnosed diabetes and each SubD grouping (ssD: OR=1.82, CIs 1.33, 2.47; mD: OR=1.95, CIs 1.39, 2.74) compared with respondents having no diabetes. No association was found between depression and undiagnosed diabetes or prediabetes compared with those having no diabetes. Milder forms of depression such as ssD and mD are more extant than major depressive episodes among adults with diabetes. The odds that an adult with diagnosed diabetes meets the criteria for ssD or mD are higher by 80% and 95%, respectively, after controlling for age, sex, race and ethnicity, education, body mass index, and poverty factors when compared against adults with no diabetes.


Increased sensitivity of carnitine palmitoyltransferase I activity to malonyl-CoA inhibition after preincubation of intact rat liver mitochondria with micromolar concentrations of malonyl-CoA in vitro.
Carnitine palmitoyltransferase I in rat liver mitochondria preincubated with malonyl-CoA was more sensitive to inhibition by malonyl-CoA than was the enzyme in mitochondria preincubated in the absence of malonyl-CoA. For carnitine palmitoyltransferase I in mitochondria from starved animals this increase also resulted in the enzyme becoming significantly more sensitive than that in mitochondria assayed immediately after their isolation. Concentrations of malonyl-CoA that induced half the maximal degree of sensitization observed were 1-3 microM.


Expression of MMP2, MMP9 and MMP3 in breast cancer brain metastasis in a rat model.
In order to study the expression of MMP2, MMP3 and MMP9 in breast cancer brain metastasis, we used a syngeneic rat model of distant metastasis of ENU1564, a carcinogen-induced mammary adenocarcinoma cell line. At six weeks post inoculation we observed development of micro-metastasis in the brain. Immunohistochemistry and Western Blotting analyses showed that MMP-2, -3 and -9 proteins expressions are consistently significantly higher in neoplastic brain tissue compared to normal brain tissue. These results were confirmed by RT-PCR. In situ zymography revealed gelatinase activity within the brain metastasis. Gel zymography showed increase in MMP2 and MMP3 activity in brain metastasis. Furthermore, we were able to significantly decrease the development of breast cancer brain metastasis in animals by treatment with PD 166793, a selective synthetic MMP inhibitor. In addition, PD 166793 decreased the in vitro invasive cell behavior of ENU1546. Together our results suggest that MMP-2, -3 and -9 may be involved in the process of metastasis of breast cancer to the brain.


Separation of thiols as phenyl mercury derivatives by thin-layer chromatography. I. Azathioprine and 6-mercaptopurine metabolites.
A method is described for converting thiol metabolites of azathioprine and 6-mercaptopurine into phenyl mercury derivatives. Separation of these derivatives was performed by chromatography on silica thin layers and they were detected by low temperature (--196 degrees) fluorescence. The parent compounds were regenerated on the chromatogram by spraying with 2 N HC1 and these were detected by low temperature fluorescence. Mercury was also detected in spots by spraying with dithizone. The method provides a simple solution to the problem of oxidation of thiol compounds during isolation procedures.


Amsacrine treatment of patients with supraventricular arrhythmias and acute leukemia.
Three patients with a history of supraventricular arrhythmia presented with relapse of acute leukemia. Two of the three patients were in sinus rhythm, receiving digoxin and/or verapamil daily. The third patient was in atrial fibrillation, but her heart rate was controlled with daily digoxin. All three patients received amsacrine without the occurrence of cardiac events. Although amsacrine may cause ventricular arrhythmias in the setting of hypokalemia, correction of the electrolyte abnormality permits its use in patients with a history of supraventricular arrhythmias.


IgG4-Related Lung Disease - Three Untreated Cases with a Benign Outcome.
IgG4-related disease is a fibroinflammatory disease in which the organs involved share similar pathological findings. Chest disease has been recently clinically and radiologically characterized. Most reports advocate prompt immunosuppressive therapy and describe a fast and good response. We report 3 cases of untreated IgG4-related lung disease that on follow-up have been clinically asymptomatic and radiologically stable or improved. In some cases of IgG4-related lung disease immunosuppressive therapy may not be warranted.


A pooled comparison of cefdinir and penicillin in the treatment of group a beta-hemolytic streptococcal pharyngotonsillitis.
This pooled analysis compared the clinical cure and bacterial eradication rates achieved by cefdinir and penicillin in the treatment of group A beta-hemolytic streptococcal (GABHS) pharngotonsillitis. Data from 4 multicenter, randomized, controlled, investigator-blinded trials, 2 in children receiving oral suspensions and 2 in adults receiving capsules/tablets, were pooled and analyzed in terms of clinical cure rates, microbiologic eradication rates, and adverse events. A total of 2751 patients were enrolled (age <13 years, n = 1274; age > or =13 years, n = 1477). Patients were randomized to receive cefdinir once daily (n = 569) or twice daily (n = 1086) for 5 or 10 days, or penicillin 4 times daily (n = 1096) for 10 days. Of the 2751 patients enrolled, 2198 were evaluable for clinical and microbiologic outcomes. Compared with the 10-day penicillin regimens, the cefdinir regimens for 5 or 10 days produced higher clinical cure and microbiologic eradication rates. Combined clinical cure rates were significantly higher for cefdinir compared with penicillin (94% vs 83%, respectively; P < 0.001). Combined microbiologic eradication rates were also significantly higher for cefdinir compared with penicillin (92% vs 77%; P < 0.001). Both cefdinir and penicillin were well tolerated, as >98% of patients completed the course of therapy. In this pooled analysis of data from 4 multicenter, randomized, controlled, investigator-blinded trials in children and adults, 5- and 10-day regimens of cefdinir achieved significantly higher clinical cure and microbiologic eradication rates compared with 10-day penicillin regimens in the treatment of GABHS pharyngotonsillitis.


Activation of purified 3-hydroxy-3-methylglutaryl-CoA reductase by phospholipids.
3-Hydroxy-3-methylglutaryl-CoA reductase (HMG-CoA reductase), the enzyme that catalyzes the rate-limiting step in cholesterol biosynthesis, has been purified by two previously reported procedures. Enzyme purified by the method of Heller, R. and Shrewsbury, M. (1976) J. Biol. Chem. 251, 3815-3822) shows up to 3-fold enhancement of activity by various types of lipid dispersions while the enzyme purified by the procedure of Tormanen et al. ((1976) Biochem. Biophys. Res. Commun. 68, 754-762) shows no activation. These results suggest that interaction with microsomal membrane lipids may be important in determining the activity of this enzyme. Analysis of bound lipid showed that enzyme prepared by the procedure of Tormanen contained at last 50 times as much phospholipid on a weight basis as enzyme prepared by Heller and Shrewsbury. Analysis of both preparations by gel-electrophoresis indicates that enzyme activities of the two comigrate, but in neither case does activity coincide with the major protein species.


Pollution patterns and their effects on biota within lotic and lentic freshwater ecosystems: How well contamination and response indicators correspond?
Aquatic environments are often severely polluted with chemical substances of anthropogenic origin, which can pose a potential threat to aquatic organisms and human health. In this study, patterns and sources of heavy metals (HMs, 6 metals) and polycyclic aromatic hydrocarbons (PAHs, 16 hydrocarbons), contamination indicators, environmental genotoxicity measures and metrics of ecological status in lotic and lentic ecosystems were collated for the first time. Chemical analysis has confirmed previously reported long-term contamination at certain study sites. The sediments of Lake Talkša, located in a city and characterized by exclusive anthropogenic pressure, exhibited the highest levels of contamination by both HMs and PAHs. Through positive matrix factorization (PMF) analysis, vehicle and industrial emissions were identified as the primary sources of HMs and PAHs. Our results revealed that frequencies of genotoxic aberrations were higher in river sites compared to lakes, with the highest genotoxic risk observed in the Nemunas River below industrial cities Alytus and Kaunas. Surprisingly, even the severely contaminated Lake Talkša showed only a "moderate" grade of genotoxic risk, highlighting the potential for adaptation of biota to long-term contamination especially in lentic ecosystems. The ecological quality status assessed by macroinvertebrate metrics, which may be sensitive to observed high biological contamination, appeared to be unrelated to contamination patterns. Consequently, to obtain the robust information on anthropogenic contamination and its effects, a combination of various assessment methods and metrics should be employed.


Sociocultural drivers of mycological knowledge: insights from Wixarika and Mestizo groups in western Mexico.
Traditional mycological knowledge (TMK) is complex, not distributed equally among the entire population, and constantly adapting to current social situations. There are sociocultural factors that could influence the fact that some people retain a greater wealth of knowledge, for instance, cultural affiliation, migration, occupation, level of schooling, and person's age. We analyze the distribution of the TMK based on sociocultural variables and 12 indicators to quantify the TMK based on a literature review. We chose two sites where there was a Wixarika and Mestizo population with records of use and consumption of wild mushrooms. In each site, 150 semi-structured interviews were conducted. The format of the semi-structured interviews was made up of sociocultural questions plus 12 questions corresponding to each of the indicators. With the data obtained, we performed linear regression tests and principal components analysis (PCA); furthermore, the significance of the groupings obtained by PCA was tested with a discriminant function analysis. We find that TMK was determined by the cultural group to which a person belongs. Contrary to what was expected, age and formal schooling did not influence people's level of knowledge. Likewise, migration and occupation were not determining factors either, although in some specific cases they did influence the differences in knowledge about mushrooms between people. The indicators that most helped to differentiate between the Wixarika people, and the Mestizos were knowledge of the nutritional contribution, propagation methods, and knowledge about toxic mushrooms. In general, sociocultural differences did not affect the transmission of the TMK due to the valorization of this knowledge among the young generations and the maintenance of the use of wild resources. Specifically, the Wixaritari had and preserved a greater TMK thanks to their pride in their cultural identity, which had allowed them to adapt to modernity while preserving their traditions and knowledge. On the other hand, the Mestizos increasingly disused wild resources due to urbanization. The indicators proposed here provided a good tool to quantify TMK; however, to replicate the study in other sites it is necessary to adapt the indicators to the context of the place.


Sexual anomalies in the estuarine snail Ilyanassa obsoleta: imposex in females and associated phenomena in males.
The spatial distribution of the "imposex" phenomenon (the imposition of male characters on females) among populations of Ilyanassa obsoleta in southern Delaware, USA estuaries was studied. Imposex was most frequent among females in a segment of the Lewes and Rehoboth Canal (68%, n=376) and there was significant spatial heterogeneity of imposex frequencies (range=43-98%) even within this habitat. Two lines of evidence indicate that conditions which generated female imposex (organotin molecules in the water) also affected male sexuality: 1) healthy males normally lose the penis following reproduction but they tended to retain it where imposex-inducing conditions were strong; and 2) males parasitized by trematodes usually lose the penis but tended not to under strong imposex-inducing conditions. The altered biology associated with imposexinducing environments extends beyond female I. obsoleta to males and to other species. This must be recognized when studying organisms exposed to these conditions.


Hypernatremic dehydration with hemorrhage into the choroid plexus.
A 2 1/2-month-old patient is reported with computed tomographic evidence of bilateral choroid plexus hemorrhage associated with hypernatremic dehydration. Choroid plexus hemorrhage may have caused increased cerebrospinal fluid protein observed in hypernatremic dehydration, as well as intraventricular hemorrhage.


Nibbling at CRF receptor control of feeding and gastrocolonic motility.
Inadequate pharmacological tools, until recently, hindered the understanding of the roles of corticotropin-releasing factor (CRF) receptor subtypes in appetite regulation and gastrocolonic motor function. Now, novel ligands that are selective for CRF(1) or CRF(2) receptors are helping to uncover the specific functions of CRF receptor subtypes. Central or peripheral CRF(2) receptor activation suppresses feeding independently of CRF(1) receptors. In the rat, central administration of CRF(2) receptor agonists promotes satiation without eliciting the malaise, behavioral arousal or anxiogenesis associated with CRF(1) receptor agonists. Conversely, central administration of CRF(1) receptor agonists elicits short-onset anorexia independently of CRF(2) receptor activation. With respect to gastrointestinal motor function, stress inhibits gastric motility through CRF(2) receptor-dependent central autonomic and peripheral myenteric systems. By contrast, stress stimulates colonic motility via CRF(1) receptor-dependent sacral parasympathetic and colonic myenteric mechanisms. These findings have important physiological implications and suggest targeted approaches for the pharmacotherapy of obesity and stress-related functional gastrointestinal and eating disorders.


Hallux valgus and metatarsus adductus measurements: inter-reader reliability and correlations on radiographs and MRI.
To assess inter-reader reliability of metatarsus adductus (MA) using the traditional method and Engel's angle (EA) on radiography and magnetic resonance imaging (MRI) and assess correlations with hallux valgus (HV). Ninety consecutive patients with radiographs and MRI of the foot were included. Two readers measured HV angle (HVA), traditional metatarsus adductus angle (MAA), and EA on radiographs and HVA and EA on MRI. Three- and two-way mixed model analyses were used for reader agreements. Ninety-five percent bootstrap confidence intervals were calculated. The linear mixed model was used for association between HVA and EA/MAA. Mean age and male to female ratio was 54.2±15.4 and 0.4:1, respectively. Mean HVA and EA were 20.6±9.4 and 21.2±8, 21.2±8.3 and 22.4±7.5 on radiographs and MRI, respectively. Mean MAA was 18.5±5.7 on radiographs. Inter-reader agreement was good for EA (ICC=0.73, 0.6) and moderate for MAA (ICC=0.41). Positive correlations between HVA, MAA, and EA on radiographs and MRI were found, but none were statistically significant (p=0.44 and 0.87). Engel's angle is more reproducible. Although positive correlations exist between the degrees of HV and MA, they are not statistically significant.


Iatrogenic alterations in radionuclide biodistributions.
The biodistribution of radiotracers used in diagnostic imaging is grossly and recognizably altered by a wide variety of drugs and other treatment modalities, such as surgery and radiotherapy. Knowledge of such altered biodistribution is important both in making diagnostic inferences from scans and in dosimetric considerations.


MARKET EVIDENCE OF MISPERCEIVED MORTALITY RISK.
We construct and implement a test of rational consumer behavior in a high-stakes financial market. In particular, we test whether consumers make systematic mistakes in perceiving their mortality risks. We implement this test using data from secondary life insurance markets where consumers with a life-threatening illness sell their life insurance policies to firms in return for an up-front payment. We compare predictions from two models: one with consumers who correctly perceive their mortality risk, and one with consumers who are misguided about their life expectancy, and find that our data are most consistent with the predictions made by the second model.


The complex interplay between stress and bacterial infections in animals.
Over the past decade, an increasing awareness has arisen of the role of neuroendocrine hormones in the susceptibility of mammalian hosts to a bacterial infection. During a stress response, glucocorticoids, catecholamines and neuroendocrine factors are released into the circulation of the host. For a long time the effects of stress on the course of an infection have been exclusively ascribed to the direct effect of stress-related hormones on the immune system and the intestinal barrier function. Chronic stress is known to cause a shift from T helper 1-mediated cellular immunity toward T helper 2-mediated humoral immunity, which can influence the course of an infection and/or the susceptibility to a microorganism. Bacteria can however also respond directly to stress-related host signals. Catecholamines can alter growth, motility, biofilm formation and/or virulence of pathogens and commensal bacteria, and as a consequence influence the outcome of infections by these bacteria in many hosts. For some bacteria, such as Salmonella, Escherichia coli and Pseudomonas aeruginosa it was shown that this influence is regulated by quorum sensing mechanisms. In this manuscript an overview of how and when stress influences the outcome of bacterial infections in animals is provided.


Use of Wisteria floribunda agglutinin affinity chromatography in the structural analysis of the bovine lactoferrin N-linked glycosylation.
Over the years, the N-glycosylation of both human and bovine lactoferrin (LF) has been studied extensively, however not all aspects have been studied in as much detail. Typically, the bovine LF complex-type N-glycans include certain epitopes, not found in human LF N-glycans, i.e. Gal(α1-3)Gal(β1-4)GlcNAc (αGal), GalNAc(β1-4)GlcNAc (LacdiNAc), and N-glycolylneuraminic acid (Neu5Gc). The combined presence of complex-type N-glycans, with αGal, LacdiNAc, LacNAc [Gal(β1-4)GlcNAc], Neu5Ac (N-acetylneuraminic acid), and Neu5Gc epitopes, and oligomannose-type N-glycans complicates the high-throughput analysis of such N-glycoprofiles highly. For the structural analysis of enzymatically released N-glycan pools, containing both LacNAc and LacdiNAc epitopes, a prefractionation protocol based on Wisteria floribunda agglutinin affinity chromatography was developed. The sub pools were analysed by MALDI-TOF-MS and HPLC-FD profiling, including sequential exoglycosidase treatments. This protocol separates the N-glycan pool into three sub pools, with (1) free of LacdiNAc epitopes, (2) containing LacdiNAc epitopes, partially shielded by sialic acid, and (3) containing LacdiNAc epitopes, without shielding by sialic acid. Structural analysis by MALDI-TOF-MS and HPLC-FD showed a complex pattern of oligomannose-, hybrid-, and complex-type di-antennary structures, both with, and without LacdiNAc, αGal and sialic acid. Applying the approach to bovine LF has led to a more detailed N-glycome pattern, including LacdiNAc, αGal, and Neu5Gc epitopes, than was shown in previous studies. Bovine milk proteins contain glycosylation patterns that are absent in human milk proteins; particularly, the LacdiNAc epitope is abundant. Analysis of bovine milk serum proteins is therefore excessively complicated. The presented sub fractionation protocol allows a thorough analysis of the full scope of bovine milk protein glycosylation. This article is part of a Special Issue entitled Glycoproteomics.


The glutathione-related detoxification pathway in the human breast: a highly coordinated system disrupted in the tumour tissues.
Glutathione and the associated enzymes, glutathione S-transferases, peroxidases, and reductase, have been implicated in cancer chemoresistance. This pathway was investigated in paired cancerous and peritumoral breast samples from 41 women. The tumours exhibited a higher redox status as deduced from increased transferase, peroxidase, and reductase activities and from higher total and reduced glutathione contents. Several components were strongly correlated in peritumoral tissues, suggesting a highly co-ordinated glutathione pathway that appeared disrupted in breast tumours with only a few correlations left. Therefore, resistance could spontaneously result from deregulated variations in the glutathione pathway, which might be relevant to the malignant disease progression.


Genetic analysis of a bile salt hydrolase in Bifidobacterium animalis subsp. lactis KL612.
To clone, sequence and characterize a new bile salt hydrolase from a bile tolerant strain of Bifidobacterium animalis ssp. lactis KL612, and further analysis of the bsh promoter and an operon-like structure containing the bsh gene in the genus Bifidobacterium. A new type of bile salt hydrolase from a bile tolerant strain of Bifidobacterium was cloned, completely sequenced and characterized. The putative bsh promoter sequence was analysed by primer extension to determine the transcriptional start point by applying the genomic walking-PCR, an operon-like structure containing the bsh gene and two more open reading frames located within a complete set ranging from a promoter to a transcription terminator sequence is reported for the first time in the genus Bifidobacterium. The polycistronic bsh transcript was revealed by reverse transcriptase-PCR (RT-PCR) as well as by Northern hybridization. Most of bile tolerant strains of bifidobacteria showed a similar genetic organization around the bsh gene. This finding suggests that bile tolerance of those strains is possibly because of the bile salt hydrolase and some transporter proteins, which are functionally related to each other to respond efficiently to the stress from bile salts. Knowledge gained through BSH research would provide further insight into the survival of probiotics in the gastrointestinal tract and some physiological functions of this enzyme in relation to the host as well as the enzyme-producing bacteria.


A histochemical reporter system to study extracellular ATP response in plants.
When cells experience acute mechanical distress, they release ATP from their cellular compartment into the surrounding microenvironment. This extracellular ATP (eATP) can then act as a danger signal-signaling cellular damage. In plants, cells adjacent to damage detect rising eATP concentrations through the cell-surface receptor kinase, P2K1. Following eATP perception, P2K1 initiates a signaling cascade mobilizing plant defense. Recent transcriptome analysis revealed a profile of eATP-induced genes sharing pathogen- and wound-response hallmarks-consistent with a working model for eATP as a defense-mobilizing danger signal. To build on the transcriptional footprint and broaden our understanding of dynamic eATP signaling responses in plants, we aimed to i) generate a visual toolkit for eATP-inducible marker genes using a β-glucuronidase (GUS) reporter system and ii) evaluate the spatiotemporal response of these genes to eATP in plant tissues. Here, we demonstrate that the promoter activities of five genes, ATPR1, ATPR2, TAT3, WRKY46, and CNGC19, were highly sensitive to eATP in the primary root meristem and elongation zones with maximal responses at 2 h after treatment. These results suggest the primary root tip as a hub to study eATP-signaling activity and provide a proof-of-concept toward using these reporters to further dissect eATP and damage signaling in plants.


Vasovasostomy: evaluation of four surgical techniques.
Four techniques of vasovasostomy were evaluated in two groups of twelve dogs each. In group 1, one side was anastomosed by a standard technique over a Silastic stent and the other side was anastomosed using a two-layer microsurgical technique. In group 2, one side was anastomosed by a standard technique without a stent and the other side was anastomosed using a one-layer microsurgical technique. Patency was evaluated by methylene blue injection distal to the anastomosis and was found to be about the same for all three groups in which anastomosis was performed without stents. The stented group had a high rate of obstruction. Histologic evaluation, however, showed superiority of both microsurgical techniques over each of the stnadard procedures.


Analysis of multiple gene co-expression networks to discover interactions favoring CFTR biogenesis and ΔF508-CFTR rescue.
We previously reported that expression of a miR-138 mimic or knockdown of SIN3A in primary cultures of cystic fibrosis (CF) airway epithelia increased ΔF508-CFTR mRNA and protein levels, and partially restored CFTR-dependent chloride transport. Global mRNA transcript profiling in ΔF508-CFBE cells treated with miR-138 mimic or SIN3A siRNA identified two genes, SYVN1 and NEDD8, whose inhibition significantly increased ΔF508-CFTR trafficking, maturation, and function. Little is known regarding the dynamic changes in the CFTR gene network during such rescue events. We hypothesized that analysis of condition-specific gene networks from transcriptomic data characterizing ΔF508-CFTR rescue could help identify dynamic gene modules associated with CFTR biogenesis. We applied a computational method, termed M-module, to analyze multiple gene networks, each of which exhibited differential activity compared to a baseline condition. In doing so, we identified both unique and shared gene pathways across multiple differential networks. To construct differential networks, gene expression data from CFBE cells were divided into three groups: (1) siRNA inhibition of NEDD8 and SYVN1; (2) miR-138 mimic and SIN3A siRNA; and (3) temperature (27 °C for 24 h, 40 °C for 24 h, and 27 °C for 24 h followed by 40 °C for 24 h). Interrogation of individual networks (e.g., NEDD8/SYVN1 network), combinations of two networks (e.g., NEDD8/SYVN1 + temperature networks), and all three networks yielded sets of 1-modules, 2-modules, and 3-modules, respectively. Gene ontology analysis revealed significant enrichment of dynamic modules in pathways including translation, protein metabolic/catabolic processes, protein complex assembly, and endocytosis. Candidate CFTR effectors identified in the analysis included CHURC1, GZF1, and RPL15, and siRNA-mediated knockdown of these genes partially restored CFTR-dependent transepithelial chloride current to ΔF508-CFBE cells. The ability of the M-module to identify dynamic modules involved in ΔF508 rescue provides a novel approach for studying CFTR biogenesis and identifying candidate suppressors of ΔF508.


Integration between anticipatory blocking and redox signaling by the peroxiredoxin/thioredoxin/thioredoxin-reductase system.
Cells are occasionally exposed to high H2O2 concentrations, often preceding exposure to other electrophylic compounds. Both H2O2 and these compounds can irreversibly modify protein thiols, with deleterious consequences. Induction of enzymatic defenses against those agents is too slow to avoid significant damage. Cells may solve this conundrum by reversibly "blocking" the thiols once H2O2 concentrations begin to increase. We term this mechanism "anticipatory blocking" because it acts in anticipation of irreversible damage upon detection of early signs of stress. Here we examine the design requirements for the Peroxiredoxin/Thioredoxin/Thioredoxin-Reductase/Protein-Dithiol System (PTTRDS) to effectively integrate H2O2 signaling and anticipatory blocking of protein dithiols as disulfides, and we compared them to the designs found in cells. To that effect, we developed a minimal model of the PTTRDS, and we defined a set of quantitative performance criteria that embody the requirements for (a) efficient scavenging capacity, (b) low NADPH consumption, (c) effective signal propagation, and (d) effective anticipatory blocking. We then sought the design principles (relationships among rate constants and species concentrations) that warrant fulfillment of all these criteria. Experimental data indicates that the design of the PTTRDS in human erythrocytes fulfills these principles and thus accomplishes effective integration between anticipatory blocking, antioxidant protection and redox signaling. A more general analysis suggests that the same principles hold in a wide variety of cell types and organisms. We acknowledge grants PEst-C/SAU/LA0001/2013-2014, PEst-OE/QUI/UI0612/2013, FCOMP-01-0124-FEDER-020978 (PTDC/QUI-BIQ/119657/2010) financed by FEDER through the "Programa Operacional Factores de Competitividade, COMPETE" and by national funds through "FCT, Fundação para a Ciência e a Tecnologia".


Using evoked compound action potentials to quantify differential neural activation with burst and conventional, 40 Hz spinal cord stimulation in ovines.
Unlike conventional dorsal spinal cord stimulation (SCS)-which uses single pulses at a fixed rate-burst SCS uses a fixed-rate, five-pulse stimuli cluster as a treatment for chronic pain; mechanistic explanations suggest burst SCS differentially modulate the medial and lateral pain pathways vs conventional SCS. Neural activation differences between burst and conventional SCS are quantifiable with the spinal-evoked compound action potential (ECAP), an electrical measure of synchronous neural activation. We implanted 7 sheep with a dorsal stimulation lead at T9/T10, a dorsal ECAP sensing lead at T6/T7, and a lead also at T9/T10 but adjacent to the anterolateral system (ALS). Both burst and conventional SCS with stimulation amplitudes up to the visual motor threshold (vMT) were delivered to 3 different dorsal spinal locations, and ECAP thresholds (ECAPTs) were calculated for all combinations. Then, changes in ALS activation were assessed with both types of SCS. Evoked compound action potential thresholds and vMTs were significantly higher (P < 0.05) with conventional vs burst SCS, with no statistical difference (P > 0.05) among stimulation sites. However, the vMT-ECAPT window (a proxy for the useable therapeutic dosing range) was significantly wider (P < 0.05) with conventional vs burst SCS. No significant difference (P > 0.05) in ALS activation was noted between conventional and burst SCS. When dosed equivalently, no differentially unique change in ALS activation results with burst SCS vs conventional SCS; in addition, sub-ECAPT burst SCS results in no discernable excitability changes in the neural pathways feeding pain relevant supraspinal sites.


Bilateral cardiac myxomas and peripheral myxomas in a patient with recent myocardial infarction.
Coronary arteriography, performed for recent anterior wall myocardial infarction in a 29 year old man, showed multiple aneurysms of the distal branches of the left coronary artery without evidence of obstructions in the major branches, and two clusters of abnormal vessels, supplying tumors in the right ventricle and the left atrium. At operation and histologic examination both tumors appeared to be myxomas. In addition, two myxomas, located elswhere in the body, could be proven histologically.


Three new orosomucoid (ORM) variants revealed by isoelectric focusing and print immunofixation.
Phenotypes of orosomucoid (ORM) in human sera have been analysed by isoelectric focusing and print immunofixation. After neuraminidase treatment the band patterns indicated that the polymorphism of the structural locus ORM1 is controlled by three autosomal codominant alleles. According to the previous nomenclature they were called ORM1F1, ORM1F2, and ORM1S. In a study of 272 unrelated individuals from southern Germany, five of the six expected common ORM1 subtypes were observed. Furthermore, we found three ORM variant phenotypes which have not been reported previously. These variants were characterized by additional bands in a cathodal position. One variant had additional double bands and presumably represents a rare ORM1 variant named ORM1S1. Two variants had additional single bands. They were assigned tentatively to the ORM2 gene locus. While the common gene product of ORM2 may be called ORM2A, the two variants are named ORM2B1 and ORM2B2, respectively. ORM2B1 has, thus far, been found only in a single individual; the variants ORM1S1 and ORM2B2 were found in a father-child pair and a mother-child pair, respectively. The frequency for variants tentatively assigned to the ORM2 locus is very low and was calculated to be 0.0037.


Childhood Maltreatment and Headache Disorders.
Childhood maltreatment is substantiated in 12 % of children, but nearly 50 % adults recall having been neglected or abused as children. Maltreatment, especially emotional abuse, is associated with migraine. Dysregulation of the HPA axis, autonomic, immune, and metabolic systems appears to be a consequence of maltreatment, and is also reported in migraine. Areas of the brain structurally and functionally affected by childhood abuse and by migraine are also similar, and include the limbic system structures, which connect to pain regions in the brainstem. Putative mechanisms by which early life stress increases the likelihood of developing migraine include gene x environment interactions, in addition to epigenetic modifications via DNA methylation. These modifications are stable and may be transferred across generations, but they may also be reversed by some medications commonly used in migraine, including valproic acid and topiramate.


Pharmacogenetics of cyclophosphamide in patients with hematological malignancies.
A high degree of interindividual variation in cyclophosphamide (CPA) pharmacokinetics was reported in certain cancer patient groups. To better understand the mechanisms underlying the variation in CPA metabolism, we have investigated the pharmacokinetics of CPA and its active metabolite 4-hydroxycyclophosphamide (4-OH-CPA) in patients with hematological tumors. The pharmacokinetics of CPA and its active metabolite were related to the genotype of CYP2B6, CYP2C9 and CYP2C19. The influence of liver function on CPA metabolism was also evaluated. Twenty-nine patients with hematological malignancies (MM, ALL or NHL) treated with a conventional CPA dose (1g/m(2)) were recruited to this study. Blood samples were collected before, during and after CPA treatment. HPLC was used to measure plasma concentrations of CPA and 4-OH-CPA. Patients were genotyped for the CYP2B6 G516T, CYP2C9*2, CYP2C9*3, CYP2C19*2 and CYP2C19*3 alleles. Serum bilirubin levels were measured before the treatment. Data was analyzed individually and by population pharmacokinetic methods, using non-linear mixed effect modeling. The interindividual variability in exposure to CPA, 4-OHCPA and 4-OH-CPA/CPA was 5.8-, 3.3- and 10.3-fold, respectively. A positive correlation between half-lives of CPA and 4-OH-CPA was found while a significant negative correlation between AUCs of CPA and 4-OH-CPA was detected. In the population analysis, the CYP2B6 G516T variant allele contribution to CPA clearance was about twice as the contribution from the wild type gene while the genotype of CYP2C9 and CYP2C19 did not influence clearance. A negative correlation was observed between bilirubin level and CPA bioactivation. This study demonstrates for the first time that the presence of the CYP2B6 G516T mutation increases the rate of 4-OH-CPA formation in patients with hematological malignancies. The liver function prior therapy as assessed by s-bilirubin influences CPA metabolism.


Normal parents: Trans pregnancy and the production of reproducers.
Some trans people want to create families in a variety of ways that include pregnancy, but often face obstacles in doing so. This paper explores how trans pregnancy is treated as exceptional and out of the ordinary by reproductive institutions. Analysis of case studies demonstrates the ubiquity of institutional obstacles to trans pregnancy and how reproductive institutions unnecessarily render trans pregnancy exceptional. Reproductive institutions shape the kinds of people for whom achieving pregnancy is made easier, and often fail to imagine the possibility of trans parents. This failure of imagination is not rooted in biological fact, but rather in social logics that ought to be the site for transformations that expand access and shift provider attitudes. Trans parents are unexceptional in the sense that, even though they may experience relatively more concentrated forms of adversity, they share many reproductive capabilities and obstacles with cis parents. In light of that concentrated adversity and the epistemic insights it might generate, how might prospective trans parents engage with new reproductive technologies? How might these engagements render them moral pioneers called to make decisions about the sorts of people created using reproductive biotechnologies?


Surgical outcome of degenerative versus postreconstructive extracranial carotid artery aneurysms.
Extracranial carotid artery aneurysms (ECAAs) are rare vascular lesions, and large series with short-term and long-term outcomes are seldom reported. This study compared the clinical presentation and conventional treatment outcomes of different ECAA types according to their etiology. We retrospectively reviewed the data of 55 consecutive patients (47 men, 8 women) with 61 ECAAs who were treated from January 1986 to December 2007 by conventional surgical techniques. The patients were a mean age of 65 +/- 11 years (range, 30-92 years). Thirty-two ECAAs (52.5%) occurred postoperatively after previous carotid endarterectomy, of which 26 patients had 29 degenerative aneurysms (47.5%). Clinical presentation included cerebral stroke in three patients (4.9%) and transient ischemic attack in 26 (42.7%). Mean follow-up was 42.7 +/- 22.0 months. Statistical analysis was performed within and between degenerative and post-reconstructive ECAA subgroups of patients. Open aneurysm resection included 27 extended polytetrafluoroethylene interposition grafts, 12 venous grafts, and 22 closures using synthetic patch. Cumulative 1-year primary patency rates were 86.9% for the degenerative ECAAs and 96% for the postoperative ECAAs, with respective secondary patency rate at 5 years of 80% and 93.3%. The 5-year patency rate was 88.9% for synthetic grafts compared with 66.7% for vein grafts and 86.4% for synthetic patches. These differences were not statistically significant (P > .05). Complications for the degenerative ECAAs included two reconstruction thromboses <30 days, two cerebral strokes, and one myocardial infarction. The patients with postoperative ECAAs experienced one early thrombosis and two strokes postoperatively. Two patients (3.6%) from the degenerative ECAA subgroup died of cardiac decompensation (n = 1) and cerebral ischemic event (n = 1). Despite the different trends, no significant differences were found between degenerative ECAA and postoperative ECAA patients in clinical presentation, localization, and surgery outcomes. The good middle-term and long-term patency rates of synthetic graft reconstruction justify its use in the treatment of ECAAs, and it is less time consuming and technically demanding compared with vein interposition graft.


Evidence that nitric oxide production increases gamma-amino butyric acid permeability of blood-brain barrier.
Blood-brain barrier permeability (BBB) to the inhibitory neurotransmitter gamma-amino butyric acid (GABA) was studied in rats following intraperitoneal (i.p) injections of GABA alone and in combination with L-Arginine (L-Arg). Administration of GABA (600 mg/kg body weight [b. wt.]) alone increased brain GABA concentration (33%, p < 0.01), when compared to untreated rats and administration of L-Arg (2000 mg/kg b. wt.) alone also increased GABA concentration (65%, p < 0.01) in the brain. Moreover, GABA + L-Arg treated brains showed a fourfold increase in GABA level (383.3%, p < 0.01) when compared to controls. Dose-dependent increase in nitric oxide production was observed 10 min after i.p injections of L-Arg (400, 800, 1000, and 2000 mg/kg b. wt.) and a peak nitric oxide (NO) production was observed at the dose level of 2000 mg/kg b. wt. On the other hand, administration of GABA failed to increase NO production in the brain. Rats pretreated (10 min) with a nonspecific nitric oxide synthase (NOS) inhibitor N(omega)-nitro-L-Arginine methyl ester (L-NAME, 50 mg/kg b. wt.) completely blocked the production of NO induced by L-Arg. In addition, L-NAME attenuated GABA entry into the brain after the administration of GABA alone or in combination with L-Arg. We conclude that high NO concentrations in the brain following L-Arg administration may increase the permeability of BBB to peripheral GABA.


Clinical significance of long non-coding RNA HOTTIP in early-stage non-small-cell lung cancer.
HOTTIP, a long non-coding RNA located in the HOXA cluster, plays a role in the patterning of tissues with mesodermal components, including the lung. Overexpression of HOXA genes, including HOTTIP, has been associated with a more aggressive phenotype in several cancers. However, the prognostic impact of HOTTIP has not yet been explored in non-small-cell lung cancer (NSCLC). We have correlated HOTTIP expression with time to relapse (TTR) and overall survival (OS) in early-stage NSCLC patients. Ninety-nine early-stage NSCLC patients who underwent surgical resection in our center from June 2007 to November 2013 were included in the study. Mean age was 66; 77.8% were males; 73.7% had stage I disease; and 55.5% had adenocarcinoma. A validation data set comprised stage I-II patients from The Cancer Genome Atlas (TCGA) Research Network. HOTTIP was expressed in all tumor samples and was overexpressed in squamous cell carcinoma (p = 0.007) and in smokers (p = 0.018). Patients with high levels of HOTTIP had shorter TTR (78.3 vs 58 months; p = 0.048) and shorter OS (81.2 vs 61 months; p = 0.023) than those with low levels. In the multivariate analysis, HOTTIP emerged as an independent prognostic marker for TTR (OR: 2.05, 95%CI: 1-4.2; p = 0.05), and for OS (OR: 2.31, 95%CI: 1.04-5.1; p = 0.04). HOTTIP was validated as a prognostic marker for OS in the TCGA adenocarcinoma cohort (p = 0.025). Moreover, we identified a 1203-mRNA and a 61-miRNA signature that correlated with HOTTIP expression. The lncRNA HOTTIP can be considered a prognostic biomarker in early-stage NSCLC.


The evolving role of reirradiation in the management of recurrent brain tumors.
Despite aggressive management consisting of surgery, radiation therapy (RT), and systemic therapy given alone or in combination, a significant proportion of patients with brain tumors will experience tumor recurrence. For these patients, no standard of care exists and management of either primary or metastatic recurrent tumors remains challenging.Advances in imaging and RT technology have enabled more precise tumor localization and dose delivery, leading to a reduction in the volume of health brain tissue exposed to high radiation doses. Radiation techniques have evolved from three-dimensional (3-D) conformal RT to the development of sophisticated techniques, including intensity modulated radiation therapy (IMRT), volumetric arc therapy (VMAT), and stereotactic techniques, either stereotactic radiosurgery (SRS) or stereotactic radiotherapy (SRT). Several studies have suggested that a second course of RT is a feasible treatment option in patients with a recurrent tumor; however, survival benefit and treatment related toxicity of reirradiation, given alone or in combination with other focal or systemic therapies, remain a controversial issue.We provide a critical overview of the current clinical status and technical challenges of reirradiation in patients with both recurrent primary brain tumors, such as gliomas, ependymomas, medulloblastomas, and meningiomas, and brain metastases. Relevant clinical questions such as the appropriate radiation technique and patient selection, the optimal radiation dose and fractionation, tolerance of the brain to a second course of RT, and the risk of adverse radiation effects have been critically discussed.


Therapy evaluation of laryngeal carcinomas by tyrosine-pet.
One of the major problems in head and neck oncology is determination of tumor status after radiotherapy. Physical examination and conventional imaging by CT and MRI do not always accurately differentiate between residual or recurrent tumor and posttreatment inflammation, fibrosis, edema, or scarring. The feasibility of positron emission tomography (PET) with L-[1-(11)C]-tyrosine (TYR) for therapy evaluation of laryngeal squamous cell carcinomas by identification of residual or recurrent disease after radiotherapy was investigated. Nineteen patients with laryngeal carcinomas had standard workups with endoscopy and conventional imaging. All subjects underwent a TYR PET scan (PET1) before definitive treatment. For determination of tumor status, a second TYR PET scan (PET2) was performed 3 months after radiotherapy. At the time of scanning, seven patients were clinically suspected of having residual disease, and in these cases, additional CT imaging and biopsies during endoscopy were performed. During the minimal follow-up period of 29 months, six patients had clinical suspicion of recurrent disease. In these six cases, a third TYR PET (PET3), CT imaging, and biopsy were performed. All pretreatment tumors were depicted by TYR PET (PET1). Three months after radiotherapy, sensitivity and specificity of TYR PET (PET2) for discrimination between residual tumor and benign posttreatment tissue changes were both 100%, and for CT, 50% and 67%, respectively. For detection of recurrent tumor during follow-up, sensitivity and specificity of TYR PET (PET3) were also 100%, and CT, 75% and 50%, respectively. Dynamic TYR PET is an accurate imaging modality for therapy evaluation in detection of residual and recurrent disease with higher sensitivity (100%) and specificity (100%) for discrimination of tumor status by TYR PET compared with conventional imaging.


Predicting Hepatocellular Carcinoma With Minimal Features From Electronic Health Records: Development of a Deep Learning Model.
Hepatocellular carcinoma (HCC), usually known as hepatoma, is the third leading cause of cancer mortality globally. Early detection of HCC helps in its treatment and increases survival rates. The aim of this study is to develop a deep learning model, using the trend and severity of each medical event from the electronic health record to accurately predict the patients who will be diagnosed with HCC in 1 year. Patients with HCC were screened out from the National Health Insurance Research Database of Taiwan between 1999 and 2013. To be included, the patients with HCC had to register as patients with cancer in the catastrophic illness file and had to be diagnosed as a patient with HCC in an inpatient admission. The control cases (non-HCC patients) were randomly sampled from the same database. We used age, gender, diagnosis code, drug code, and time information as the input variables of a convolution neural network model to predict those patients with HCC. We also inspected the highly weighted variables in the model and compared them to their odds ratio at HCC to understand how the predictive model works. We included 47,945 individuals, 9553 of whom were patients with HCC. The area under the receiver operating curve (AUROC) of the model for predicting HCC risk 1 year in advance was 0.94 (95% CI 0.937-0.943), with a sensitivity of 0.869 and a specificity 0.865. The AUROC for predicting HCC patients 7 days, 6 months, 1 year, 2 years, and 3 years early were 0.96, 0.94, 0.94, 0.91, and 0.91, respectively. The findings of this study show that the convolutional neural network model has immense potential to predict the risk of HCC 1 year in advance with minimal features available in the electronic health records.


Mapping DNA methylation across development, genotype and schizophrenia in the human frontal cortex.
DNA methylation (DNAm) is important in brain development and is potentially important in schizophrenia. We characterized DNAm in prefrontal cortex from 335 non-psychiatric controls across the lifespan and 191 patients with schizophrenia and identified widespread changes in the transition from prenatal to postnatal life. These DNAm changes manifest in the transcriptome, correlate strongly with a shifting cellular landscape and overlap regions of genetic risk for schizophrenia. A quarter of published genome-wide association studies (GWAS)-suggestive loci (4,208 of 15,930, P < 10(-100)) manifest as significant methylation quantitative trait loci (meQTLs), including 59.6% of GWAS-positive schizophrenia loci. We identified 2,104 CpGs that differ between schizophrenia patients and controls that were enriched for genes related to development and neurodifferentiation. The schizophrenia-associated CpGs strongly correlate with changes related to the prenatal-postnatal transition and show slight enrichment for GWAS risk loci while not corresponding to CpGs differentiating adolescence from later adult life. These data implicate an epigenetic component to the developmental origins of this disorder.


Chest wall implantation of lung cancer after thin-needle aspiration biopsy.
Percutaneous aspirational biopsy of intrathoracic lesions has become a popular technique. One of the rarest complications is that of implantation of malignant cells within the needle tract and subsequent development of a chest wall mass. A case of chest wall implantation of adenocarcinoma of the lung after thin-needle biopsy is reported. The lesion was successfully treated with radical full-thickness excision of the chest wall and immediate reconstruction with a large rectus abdominis musculocutaneous flap.


Inhibition of ICAM-1/LFA-1-mediated heterotypic T-cell adhesion to epithelial cells: design of ICAM-1 cyclic peptides.
In this work, we have designed cyclic peptides (cIBL, cIBR, cIBC, CH4 and CH7) derived from the parent IB peptide (ICAM-1(1-21)) that are inhibitors of ICAM-1/LFA-1-mediated T-cell adhesion to Caco-2 cell monolayers. Cyclic peptide cIBR has the best activity of any of the peptides evaluated. The active ICAM-1 peptides have a common Pro-Arg-Gly sequence that may be important for binding to LFA-1.


Identification, expression profile and analysis of the antimicrobial activity of collectin 11 (CL-11, CL-K1), a novel complement-associated pattern recognition molecule, in half-smooth tongue sole (Cynoglossus semilaevis).
Collectin 11 (CL-11, also known as collectin kidney-1, CL-K1), a new member of the vertebrate C-type lectin superfamily, plays an important role in innate immunity as a pattern recognition molecule of the lectin complement pathway. However, little is known about CL-11 in teleosts. In the present study, a CL-11 homolog was identified and characterized from half-smooth tongue sole (Cynoglossus semilaevis) (designated as CsCL-11). The full-length cDNA of CsCL-11 is 1220 bp long and includes a 5'untranslated region (5'-UTR) of 180 bp, a 3'-UTR of 218 bp and an open reading frame (ORF) of 819 bp encoding 273 amino acids. Multiple sequence alignment revealed that the deduced CsCL-11 protein has the typical modular architecture (EPN and WTD) conserved throughout vertebrates, suggesting a conserved function of CsCL-11. Tissue expression profile analysis by quantitative real-time PCR (qRT-PCR) showed CsCL-11 to be ubiquitously distributed in tissues and highly expressed in the ovary and liver. A pattern of significant upregulation of CsCL-11 expression was observed in the blood, spleen, head kidney and gill at 6 h, 12 h and 24 h after infection with Vibrio anguillarum, and western blotting showed that natural CsCL-11 protein levels in the blood were significantly increased after V. anguillarum infection. Moreover, by binding to various bacteria, recombinant CsCL-11 (rCsCL-11) expressed in HEK-293 T cells displayed strong antibacterial activity. Taken together, these results suggest that CsCL-11 is a unique C-type lectin that is likely involved in host defense against bacterial infection. To our knowledge, this is the first study on CL-11 in marine fish.


Effects of microplastics and food particles on organic pollutants bioaccumulation in equi-fugacity and above-fugacity scenarios.
Microplastics (MPs), as emerging contaminants, sorb organic pollutants from the environment or leach out additives, thereby altering the fate of co-existing pollutants to organisms. We chose equi-fugacity and above-fugacity concentrations of polychlorinated biphenyls (PCBs) as background contamination and plastic additive concentrations, respectively, to investigate the effects of MPs on PCB bioaccumulation; we compared the effects of MPs with those of food-borne particles (FBPs). Co-exposure to MPs and FBPs at both the equi-fugacity and above-fugacity PCB concentrations had no obvious toxic effects (ROS generation and cyp1a expression) on zebrafish. When the zebrafish were exposed to the equi-fugacity PCB concentrations, the PCB concentrations reached 177.7-400.5 ng/g after a 7-d uptake; the presence of MPs did not significantly enhance PCB bioaccumulation. The remaining PCB concentrations in the fish after a 4-d depuration were 58.4-125.1 ng/g; the effects of MPs were the same as those during the uptake period. However, at the above-fugacity PCB concentrations, the MPs markedly increased the PCB bioaccumulation (by 1.8-fold) to 712.9 ng/g. This is because at above-fugacity concentrations, PCBs on MPs migrate to organisms as there were high fugacity gradients. The FBPs enhanced PCB bioaccumulation in zebrafish more effectively than the MPs, even after depuration. In the presence of FBPs, PCB bioaccumulation increased by 2.8- and 4.2- fold after uptake in the equi-fugacity and above-fugacity scenarios, respectively, both of which were significantly higher than that observed for the MPs. This is probably because FBPs are easily assimilated by fish, making the associated PCBs more bioavailable. Finally, during the co-existence of MPs and FBPs, MPs facilitate the depuration of PCBs accumulated via FBP vectors; conversely, FBPs did not affect PCB accumulation via MP vectors. Thus, this study elucidated the effects of MPs and FBPs on the bioaccumulation of pollutants at equi-fugacity or above-fugacity concentrations in aquatic environments.


Thumb metacarpophalangeal ulnar collateral ligament injuries: a biomechanical simulation study of four static reconstructions.
To determine the optimal tunnel placement positions for tendon graft reconstruction of chronic thumb metacarpophalangeal (MCP) ulnar collateral ligament injuries that would stabilize the joint while maintaining motion. Four commonly used tunnel placement methods were simulated on a cadaveric model using fresh-frozen thumbs and a suture/screw construct. The methods were as follows: (1) triangular configuration with apex proximal, (2) triangular configuration with apex distal, (3) cruciate configuration, and (4) parallel configuration. Stability was tested by valgus loading at 0 degrees and 30 degrees , and range of motion was tested by loading the thumb tendons. Statistical analysis was performed by 1-way analysis of variance testing. Valgus load stability testing at 0 degrees and 30 degrees showed that all 4 reconstruction methods stabilized the MCP joint compared with the fully sectioned state. The amount of stability achieved was not significantly different between the 4 methods. Only the reconstruction method, however, with a triangular configuration with the apex proximal restored flexion/extension range of motion not significantly different from the intact state. The other 3 methods resulted in significantly decreased range of motion. The reconstruction tunnel positioning of triangular configuration with apex proximal stabilizes the thumb MCP joint while maintaining flexion/extension range of motion. We recommend this configuration for chronic MCP joint injuries in which the native ulnar collateral ligament is inadequate and tendon graft reconstruction is performed.


Concurrent radiochemotherapy in the treatment of locally advanced non-small cell lung cancer.
Current evidence suggests that concurrent RT/CHT is the standard of treatment in patients with locally advanced, inoperable NSCLC. This treatment approach, however, bears risk of increased toxicity and, therefore, must be carefully balanced regarding the choice of patients suitable for such an intervention. Newer RT technologies, third-generation CHT drugs, and widely used radioprotectors represent the current framework for numerous clinical studies that ultimately will help optimize the treatment of this disease.


Understanding the Role of Removable Solid Additives: Selective Interaction Contributes to Vertical Component Distributions.
Sequentially deposited organic solar cells (SD-OSCs) have attracted great attention owing to their ability in achieving a more favorable, vertically phase-separated morphology to avoid the accumulation of counter charges at absorber/transporting layer interfaces. However, the processing of SD-OSCs is still quite challenging in preventing the penetration of small-molecule acceptors into the polymer donor layer via erosion or swelling. Herein, solid additives (SAs) with varied electrostatic potential distributions and steric hinderance are introduced into SD-OSCs to investigate the effect of evaporation dynamics and selective interaction on vertical component distribution. Multiple modelings indicate that the π-π interaction dominates the interactions between aromatic SAs and active layer components. Among them, p-dibromobenzene shows a stronger interaction with the donor while 2-chloronaphthalene (2-CN) interacts more preferably with acceptor. Combining the depth-dependent morphological study aided by multiple X-ray scattering methods, it is concluded that the evaporation of SAs can drive the stronger-interaction component upward to the surface, while having minor impact on the overall molecular packing. Ultimately, the 2-CN-treated devices with reduced acceptor concentration at the bottom surface deliver a high power conversion efficiency of 19.2%, demonstrating the effectiveness of applying selective interactions to improve the vertical morphology of OSCs by using SAs with proper structure.


Reassessing the role of morphology in bryophyte phylogenetics: Combined data improves phylogenetic inference despite character conflict.
Morphological data has gained renewed attention and has been shown to be crucial in clarifying the phylogenetic relationship in a wide range of taxa. In the last decades, phylogenetic analyses of sequence-level data have radically modified the systematic schemes within bryophytes (early non-vascular land plants) and have revealed a widespread pattern of conflict with morphology-based classifications. Yet, a comprehensive evaluation of character conflict has not yet been performed in the context of combined matrices. In this study, we evaluate the impact of morphology on bryophyte phylogeny following a total-evidence approach across 10 published matrices. The analysed matrices span a wide range of bryophytes, taxonomic levels, gene sampling and number of morphological characters and taxa. Data conflict was addressed by measuring: (i) the topological congruence between individual partitions, (ii) changes in support values of the combined data relative to the molecular partition and (iii) clade stability. The association between these measures and the number of morphological characters per taxon (Nc/T ratio) and the proportion of non-fixed characters (i.e., inapplicable, polymorphic and missing data) was explored. In the individual partition analyses, the Nc/T ratio correlated positively with the topological congruence in six to seven datasets depending on the weighting scheme. The proportion of non-fixed cells had a minor influence on congruence between data partitions. The number of characters and proportion of non-fixed data varied significantly between morphological datasets that improved congruence between data types. This variation suggests that morphological datasets affect the results of combined analyses in different ways, depending on the taxa studied. Combined analyses revealed that, despite the low congruence values between partitions, integrating data types improves support values and stability. However, while non-fixed data had no negative effect on support values, stability was reduced as the proportion of non-fixed cells increased. Nc/T ratio was negatively associated with support values and it showed ambiguous responses in stability evaluations. Overall, the results indicate that adding morphology may contribute to the inference of phylogenetic relationships of bryophytes despite character conflict. Our findings suggest that merely comparing (a) morphology-based classifications with molecular phylogenies or (b) the outcome from individual data partitions can misestimate data conflict. These findings imply that analyses of combined data may provide conservative assessments of data conflict and, eventually, lead to an improved sampling of morphological characters in large-scale analyses of bryophytes.


Characteristics of Highly Cited Articles in Interventional Cardiology.
Citation classics have been published in many fields of medicine; however, none have focused on interventional cardiology. The goal of this study was to identify the top 100 articles in the field of interventional cardiology and highlight their important trends and characteristics. The Scopus database was used by 2 independent reviewers to extract the top 100 articles using a variety of keywords. We found articles published between 1953 and 2012. Majority (n = 78) of the top 100 articles were published between 1996 and 2010, and the United States was affiliated with the highest number of articles in our list (n = 68). Over half (n = 54) the articles were funded. Private funding was correlated with higher citations (p = 0.036). A third (n = 33) of the papers had authors with conflicts of interest; however, conflict of interest had no effect on citations (p = 0.837). Majority (n = 57) of the articles studied coronary angioplasty and stenting; followed by coronary angiography (n = 14). Women were underrepresented, with only 11 female first authors in the top 100 papers, and only 1 female in the list of top authors who had 5 or more publications. In conclusion, the following features define the typical highly cited article in interventional cardiology-a clinical trial conducted in the United States, which studies angioplasty, and has been published relatively recently in a high-impact journal by a male first author.


Identification of rabies virus mimotopes screened from a phage display peptide library with purified dog anti-rabies virus serum IgG.
The rabies virus glycoprotein (G) is a key protein for both virus infectivity and eliciting protective immunity as an antigen. What is more, the nucleoprotein (N) is also a significant rabies virus antigen. In this study, purified anti-rabies virus IgG from dogs immunized with the standard CVS-11 strain was used to screen the Ph.D.-12™ Phage Display Peptide Library for peptides that correspond to or mimic native G and N epitopes. In contrast to previous reports that use monoclonal antibodies or human anti-rabies virus serum, this study describes the first use of dog serum to screen for epitopes. After three rounds of biopanning, selected phage clones were identified by plaque screening, western blotting (WB), and ELISA. Positive phage clones were sequenced, and their amino acid sequences were deduced. Alignment of the peptide sequences to G and N indicated that the epitope peptides matched well with G amino acids at positions 34-42, 198-200, 226-264, 296-371, and 330-343, as well as to N amino acids at positions 22-168 (N-terminal) and 262-450 (C-terminal), confirming that the sequences were indeed mimicking epitopes. Thirty percent of the selected clones matched reported antigenic regions located at sites II and III of the glycoprotein. Two sequences, LEPKGRYDDPWT and ATRYDDIWASTA, that have no homology to the known antigenic sites of either the G or N exhibited a common RYDD-W-T motif that is highly homologous to the amino acid residues at positions 126-141 of the G. This finding indicates that this motif may be a new potential RABV G B cell epitope. Amino acids 126-141 containing the RYDD-W-T motif may become a novel key epitope region and allow the development of a rabies vaccine or diagnostic reagents for the treatment of rabies.


A graphene oxide-based FRET sensor for rapid and sensitive detection of matrix metalloproteinase 2 in human serum sample.
Graphene oxide (GO) has been widely used to develop fluorescence resonance energy transfer (FRET) biosensors for tumor markers (e.g., matrix metalloproteinases, MMPs) due to its superior fluorescence quenching capacity and unique adsorption characteristics for biomolecules. In this study, fluorescein isothiocyanate-labeled peptide (Pep-FITC) was assembled onto the GO surface through covalent binding to construct a GO-Pep-FITC FRET sensor for sensitive, rapid, and accurate detection of MMP-2 in complex serum samples. Compared to similar GO-based FRET sensors fabricated through physical adsorption, the as prepared ones via covalent binding are significantly more stable under physiological conditions, enabling their detection of MMP-2 with high sensitivity (detection limit: 2.5ng/mL). More importantly, it allows for rapid MMP-2 detection (within 3h) even in complex biological samples with satisfactory accuracy and the relative standard deviation ≤7.03%. Our studies further suggest that such a platform developed here for sensitive, rapid, and accurate detection of biomarkers holds great promise for clinical diagnosis of protease-related diseases.


Action mechanism of corticosteroids to aggravate Guillain-Barré syndrome.
Corticosteroids have been proved to be ineffective for Guillain-Barré syndrome, but the mechanism remains unknown. In a rabbit model of axonal Guillain-Barré syndrome, treatment with corticosteroids significantly reduced macrophage infiltration in the spinal ventral roots and the survival rate as well as clinical improvement. On 30(th) day after onset, there was significantly higher frequency of axonal degeneration in the corticosteroids-treated rabbits than saline-treated rabbits. Corticosteroids may reduce the scavengers that play a crucial role for nerve regeneration, thus delay the recovery of this disease.


Diagnostic Accuracy of Bronchodilator Response for Asthma in a Population of South China.
A significant bronchodilator response is commonly defined as a 12% or greater and 200 ml or greater change in FEV1 from baseline according to the 2005 American Thoracic Society (ATS)/European Respiratory Society (ERS) criterion. A number of studies have shown that the ATS/ERS criterion has limitations in asthma diagnosis, and some experts have argued for correcting the criteria. The aim of this study is to investigate the diagnostic value of acute bronchodilator response for asthma in a Southern Chinese population. We prospectively evaluated 805 patients with obstructive lung disease (309 for asthma, 496 for non-asthma). Spirometry was performed according to the ATS/ERS guidelines. Data were analyzed by SPSS 18.0. The receiver-operating characteristic (ROC) curve was drawn to assess the diagnostic accuracy of the ATS/ERS criterion based on FEV1. Linear regression was used to analyze the factors of FEV1 change. The sensitivity and specificity of the acute bronchodilator test when judged by the ATS/ERS criteria (200 ml or higher and 12% improvement) were 68.6% and 78.2%, respectively. For the ATS/ERS criteria, the Youden Index, which comprehensively reflects the authenticity of a diagnostic test, was 46.8%. The absolute change of FEV1 positively correlated with baseline FEV1 and weight and negatively with age, while the percentage change of FEV1 was negatively correlated with baseline FEV1, age and height and positively with weight. Compared with the different diagnostic values, when ∆FEV1 was 195 ml and ∆FEV1i% was 14%, the Youden Index was the largest (48.2%) and the diagnostic capability of the test the biggest. The ATS/ERS criterion for acute bronchodilator response might not be completely suitable for asthma in the Chinese population. Chinese Clinical Trial Registry (Registry ID: ChiCTR-DDT-14004976). This work was supported by the National Natural Science Foundation of China (grant nos. 81670027, 81270080).


Intracellular localization of fructose 1,6-bisphosphate aldolase.
Submission of a rat liver homogenate made in 250 mM sucrose-1 mM EDTA to centrifugation between 9,500 times g for 10 min and 105,000 times g for 60 min results in the sedimentation of 60 to 70% of the total cellular fructose 1,6-bisphosphate aldolase (EC 188.8.131.52). Under these conditions only about one-quarter of the total triose phosphate dehydrogenase and phosphoglycerate kinase appears in the microsomal fraction. Ultrastructural immunologic localization techniques have demonstrated that the aldolase is associated with the endoplasmic reticulum, in situ. The binding of this enzyme to the membrane is sensitive to changes in pH with an optimum at 6.0, and to increasing concentrations of NaCl and fructose 1,6-bisphosphate, being about 100-fold more sensitive to the ester than to the inorganic salt.


Long-term morphologic fundus and optic nerve head pattern of progressive myopia in congenital glaucoma distinguished by age at first surgery.
The purpose of this study was to investigate the preservation of round optic nerve head (ONH) shape in myopic eyes of surgically treated congenital glaucoma patients, with regard to factors associated with intraocular pressure (IOP) elevation-induced peripapillary scleral (PPS) deformation. Using optical coherence tomography (OCT) on the ONH and macula, we identified myopic eyes with round ONH and internally oblique border tissue and those with non-round ONH. We investigated differences in clinical factors between the two groups. We included 51 eyes of 34 patients. Age at first surgery (2.8 vs. 15.2 months, P < 0.001) was significantly different between the two groups. Axial length was also significantly longer (P = 0.004) in the non-round group, but multiple logistic regression analysis revealed age as the only significant factor (P < 0.05) in ONH roundness. Interestingly, the round ONH group also had non-curved fundus morphology and a thick choroid, while the non-round ONH group showed diverse degrees of disc tilt and posterior pole curvature, and a thin choroid. In conclusion, in eyes with congenital glaucoma, age at first surgery, particularly when older than 6 months, was associated with round ONH and emmetropia-like fundus despite high myopia. The findings may indicate two different changes in the posterior sclera and the neural canal in response to the increased IOP.


[Early diagnosis of diabetes mellitus--studies on the effectiveness and value of diabetes screening actions].
Actions to found out cases of diabetes, which exclusively worked with a proof of glucosuria using the biophan-test stripe, were always loaded by a high percentage of falsely pathologic, but also by a not insignificant number of falsely negative investigation results. -- Therefore a multiphasic screening was carried out in 5,424 persons at the age of 20 and more years, which tested the problems mentioned and judged the possibilities of an improved diagnosis.


Effect of ethanol on cyclic nucleotides in vivo: consequences of controlling motor and respiratory changes.
Many psychotropic drugs alter cerebellar cyclic guanosine-3',5'-monophosphate (cGMP) content. Whereas apomorphine increased levels, central depressants such as ethanol, chlordiazepoxide or barbiturates, reduce the content of cerebellar cGMP without altering levels of cyclic adenosine-3',5'-monophosphate (cAMP). Additional data indicate that tolerance develops to this reduction of cerebellar cGMP by ethanol. In paralyzed animals, the increase in cerebellar cGMP content induced by apomorphine and the decrease caused by ethanol were dramatically attenuated. Since relatively high doses of ethanol were needed to decrease blood CO2 tension in spontaneously moving rats, changes in respiratory function appear to be of only minor importance in the ethanol-induced decrease in cerebellar cGMP. It is concluded that ethanol-induced changes in content of cerebellar cGMP in vivo may be secondary to alterations in motor and, to a lesser extent, in respiratory function.


PGF2 alpha, thromboxane B2 and HETE levels in gerbil brain cortex after ligation of common carotid arteries and decapitation.
The effects of ligation of both common carotid arteries in the gerbil on the levels of PGF2 alpha, TXB2, HETE and of energy metabolites in brain cortex, have been investigated. Also, in the same experimental conditions the changes of cyclic AMP in brain cortex, cerebellum, striatum and hippocampus have been monitored. ATP, glycogen, glucose and phosphocreatine decrease whereas, lactate and cyclic AMP are enhanced in the ischemic brain, as previously reported. In contrast, levels of arachidonic acid metabolites are not modified. During ischemia following decapitation, instead, PGF2 alpha, and TXB2, show considerable increase.


Relationships among aural abscesses, organochlorine compounds, and vitamin a in free-ranging eastern box turtles (Terrapene carolina carolina).
Aural abscesses are a common health problem in free-ranging eastern box turtles (Terrapene carolina carolina), and they have been associated with high body burdens of organochlorine (OC) compounds, which are known disruptors of vitamin A. The objective of this study was to determine if the presence of pathologic lesions in box turtles were correlated with increased and decreased levels of hepatic OC compounds and vitamin A, respectively. A graded scale for the pathologic changes observed in tissue samples collected from abscessed and nonabscessed box turtles over a 2-yr period (2003-04) was developed, and the levels of OC compounds and vitamin A in livers collected from the same turtles were determined through chemical analysis. Sixty-eight turtles (40 with aural abscesses and 28 without) were included in the study. Relationships between variables were analyzed using Spearman's Rank Correlation Test, where P</=0.05 was considered significant. Twenty-seven different OC compounds were identified. Mean+/-standard deviation (SD) total OC compound level for all turtles was 0.35+/-0.83 ppm (range 0-5.81 ppm), and mean+/-SD vitamin A level was 72.8+/-98.6 ppm (range 0-535.7 ppm). There was no correlation between pathologic score and total hepatic OC compound concentration (r = -0.18, P = 0.16). However, pathologic score was positively correlated with o,p'-DDT (r = 0.25, P = 0.05). Vitamin A was positively correlated with pathologic score (r = 0.32, P = 0.01), which was contrary to the expected result. There was no linear correlation between vitamin A and total hepatic OC compound concentration (r = -0.04, P = 0.75). However, a nonlinear regression provided a significant fit (r(2) = 0.12, P = 0.02), indicating an initial increase in vitamin A as the OC compound burden increased, followed by a decline as OC compound levels increased further. The hepatic OC compound residue concentrations in these box turtles were lower compared to levels found in freshwater aquatic turtles but similar to levels in other terrestrial reptile species. This study provides mixed results in support of a role of OC compounds, presumably of environmental origin, in the etiology of aural abscesses in free-ranging box turtles.


Efficacy of selamectin in the prevention of adult heartworm (Dirofilaria immitis) infection in dogs in northern Italy.
The efficacy of a novel avermectin, selamectin, was evaluated for the prevention of heartworm disease (adult Dirofilaria immitis infection) in 120 dogs (aged 9 months to 13 years at enrolment) presented as veterinary patients. The study was conducted at five veterinary practices in a heartworm hyperendemic region of northern Italy. Dogs were allocated randomly in a 2:1 ratio to treatment with either selamectin or ivermectin. Treatments were administered at monthly intervals for 6 months during the heartworm transmission season (May-November). Selamectin was applied topically in a single spot to the skin on each animal's back at the base of the neck in front of the scapulae as a unit dose that provided at least the minimum recommended dosage of 6mgkg(-1) (range, 6-12mgkg(-1)). Ivermectin (6microgkg(-1) of body weight) was administered orally at monthly intervals, in accordance with the manufacturer's product label recommendations. Study day 0 was defined individually for each dog as the day of first treatment administration. Efficacy was assessed on the basis of the absence of D. immitis microfilariae and adult heartworm (D. immitis) antigen in tests conducted on days 180 and 300. There were no adverse clinical signs arising due to treatment with selamectin and no drug-related mortalities. The prevention rate for D. immitis microfilariae and adult heartworm antigen was 100% for both selamectin and ivermectin. Thus, selamectin administered as a unit dose, providing at least the recommended minimum dosage of 6mgkg(-1), at monthly intervals during the heartworm transmission season was safe and 100% effective in the prevention of heartworm disease in dogs presented as veterinary patients.


Cognitive impairment of aphasics in a colour-to-picture matching task.
Broca's and Wernicke's aphasics, non-aphasic brain-damaged patients and chronic schizophrenics were tested in a colour-to-picture matching task. For each of 42 black-and-white drawings of objects the most "typical" colour was to be selected from a set of 24 colour strips. Separate scores for the appropriateness of the choices as to hue, brightness, and saturation were obtained by comparison with the data from 50 normal Ss. The choices of both aphasic groups were more deviant than those of brain-damaged controls for chromatically coloured objects with respect to all three colour dimensions, and for achromatically coloured objects with respect to saturation and brightness. Wherever the aphasics' performance was worse than that of the controls, the deviancy-scores correlated significantly with the Token Test. Results are interpreted as an impairment of aphasics in the ability to isolate analytically specific features of objects or concepts which might be conceived of as a prerequisite of propositional language.


Towards optimizing peptide-based inhibitors of protein-protein interactions: predictive saturation variation scanning (PreSaVS).
A simple-to-implement and experimentally validated computational workflow for sequence modification of peptide inhibitors of protein-protein interactions (PPIs) is described.


Integrative genomic analysis of PPP3R1 in Alzheimer's disease: a potential biomarker for predictive, preventive, and personalized medical approach.
Alzheimer's disease (AD) is associated with abnormal calcium signaling, a pathway regulated by the calcium-dependent protein phosphatase. This study aimed to investigate the molecular function of protein phosphatase 3 regulatory subunit B (PPP3R1) underlying AD, which may provide novel insights for the predictive diagnostics, targeted prevention, and personalization of medical services in AD by targeting PPP3R1. A total of 1860 differentially expressed genes (DEGs) from 13,049 background genes were overlapped in AD/control and PPP3R1-low/high cohorts. Based on these DEGs, six co-expression modules were constructed by weight gene correlation network analysis (WGCNA). The turquoise module had the strongest correlation with AD and low PPP3R1, in which DEGs participated in axon guidance, glutamatergic synapse, long-term potentiation (LTP), mitogen-activated protein kinase (MAPK), Ras, and hypoxia-inducible factor 1 (HIF-1) signaling pathways. Furthermore, the cross-talking pathways of PPP3R1, such as axon guidance, glutamatergic synapse, LTP, and MAPK signaling pathways, were identified in the global regulatory network. The area under the curve (AUC) analysis showed that low PPP3R1 could accurately predict the onset of AD. Therefore, our findings highlight the involvement of PPP3R1 in the pathogenesis of AD via axon guidance, glutamatergic synapse, LTP, and MAPK signaling pathways, and identify downregulation of PPP3R1 as a potential biomarker for AD treatment in the context of 3P medicine-predictive diagnostics, targeted prevention, and personalization of medical services. The online version contains supplementary material available at 10.1007/s13167-021-00261-2.


Trough oestradiol levels associated with cognitive impairment in post-menopausal women after 10 years of oestradiol implants.
Oestrogen treatment has been found to improve memory in healthy post-menopausal women, but the effects are small, especially when socio-economic status is controlled. There is some evidence that with long-term treatment the benefits decline, or reverse. To examine the effects of 10 years of oestradiol implants on attention, memory and frontal lobe function in healthy women (aged 51-72 years) who had undergone a surgical menopause. In an open study, patients were recruited from the Guy's Hospital menopause clinic and were tested just before receiving a new implant. Exclusion criteria included any form of psychoactive medication, IQ<90, English not the first language and anxiety or depression scores in the clinical range. All patients had been receiving HRT with an oestradiol implant for about 10 years. Each patient was pair-matched to a woman who had never received HRT. Eighteen pairs were matched for age, IQ, socio-economic status, years of secondary education, and occupation. They completed questionnaires, mood rating scales and a battery of cognitive tests. The patients with oestradiol implants had significantly greater psychological and somatic menopausal symptoms and significantly worse mental flexibility (rule reversal) and long-term episodic memory than did the control group who had never received HRT. Analysis of covariance showed that the difference in menopausal symptoms did not account for the cognitive differences. There were no differences in a test of sustained attention or in a category generation task or in self-ratings of mood after the stress of cognitive testing. Long-term treatment with oestradiol implants does not convey cognitive benefits and may lead to impaired function.


PAX4 enhances beta-cell differentiation of human embryonic stem cells.
Human embryonic stem cells (HESC) readily differentiate into an apparently haphazard array of cell types, corresponding to all three germ layers, when their culture conditions are altered, for example by growth in suspension as aggregates known as embryoid bodies (EBs). However, this diversity of differentiation means that the efficiency of producing any one particular cell type is inevitably low. Although pancreatic differentiation has been reported from HESC, practicable applications for the use of beta-cells derived from HESC to treat diabetes will only be possible once techniques are developed to promote efficient differentiation along the pancreatic lineages. Here, we have tested whether the transcription factor, Pax4 can be used to drive the differentiation of HESC to a beta-cell fate in vitro. We constitutively over-expressed Pax4 in HESCs by stable transfection, and used Q-PCR analysis, immunocytochemistry, ELISA, Ca(2+) microfluorimetry and cell imaging to assess the role of Pax4 in the differentiation and intracellular Ca(2+) homeostasis of beta-cells developing in embryoid bodies produced from such HESC. Cells expressing key beta-cell markers were isolated by fluorescence-activated cell sorting after staining for high zinc content using the vital dye, Newport Green. Constitutive expression of Pax4 in HESC substantially enhances their propensity to form putative beta-cells. Our findings provide a novel foundation to study the mechanism of pancreatic beta-cells differentiation during early human development and to help evaluate strategies for the generation of purified beta-cells for future clinical applications.


Identification of virally encoded microRNAs.
RNA silencing is a widespread phenomenon that regulates gene expression at different levels. Small RNA molecules are at the core of all RNA silencing pathways and can be grouped in distinct families depending on their chemical properties and their mode of action. Among these small RNAs, microRNAs (miRNAs) represent one of the most extensively studied classes in animals. These tiny endogenous RNAs regulate gene expression posttranscriptionally by cleaving the targeted transcript or by interfering with its translation. miRNAs are found in plants and animals and have been identified in mammalian viruses. This indicates that mammalian viruses exploit the host RNA silencing machinery to produce miRNAs that have the potential to act both on the infected host genome and on the viral genome. The techniques used for identification of viral miRNAs mostly parallel what is used for identifying cellular miRNAs. The use of prediction algorithms followed by validation has been successful. A more direct and nonbiased way of identifying viral miRNAs consists of cloning and sequencing small RNA libraries from virally infected cells or tissues.