Transcriptomic and iTRAQ proteomic approaches reveal novel short-term hyperosmotic stress responsive proteins in the gill of the Japanese eel (Anguilla japonica).
Osmoregulation is critical for the survival of fishes that migrate between freshwater (FW) and seawater (SW). The eel, as a catadromous fish, has been studied for decades to reveal the mechanisms of osmoregulation. These studies, however, have been limited by the lack of a genomic database to decipher the mechanism of osmoregulation at a molecular level. In this study, using high-throughput transcriptomic and proteomic technologies, we have provided the first genome-wide study to identify hyperosmotic responsive proteins in the gills of the Japanese eel. Deep sequencing using the 454 platform produced over 660,000 reads with a mean length of 385 bp. For the proteomic study, we collected gill samples from three different treatment groups of fish that had fully adapted to FW/SW or were transferred from FW to SW for 6h. The respective group of gill proteins were extracted and labeled using an isobaric tag for relative and absolute quantitation (iTRAQ) using LTQ-Orbitrap, a high resolution mass spectrometer. Among the 1519 proteins identified from the gill samples, 96 proteins were differentially expressed between FW and SW adapted fish. Nineteen hyperosmotic responsive proteins were detected (10 up-regulated and 9 down-regulated proteins) after 6h post FW to SW transfer. The study has provided the most comprehensive, targeted investigation of eel gill proteins to date, and shown the powerfulness of combining transcriptomic and proteomic approaches to provide molecular insights of osmoregulation mechanisms in a non-model organism, eel.


A historical and clinical perspective endorsing person-centered management of fibromyalgia syndrome.
Fibromyalgia or fibromyalgia syndrome (FMS) is a complex chronic pain disorder of unknown causation frequently associated with debilitating fatigue, unrefreshing sleep, cognitive and affective symptoms. A fibromyalgia-type suffering was possibly described in the Book of Job. Analogous symptomatic conditions have been medically recognized since the early 1900s, when initially labeled as "fibrositis". Since the early 1980s, FMS has evolved and differentiated after its characterization in a controlled study. Since then, research has focused on multiple aspects of this disorder, including characterization and management of symptoms, psychophysiology, neuroendocrine-immune pathophysiology, including central sensitization mechanisms. The complex and multifaceted nature of FMS lends itself better to a holistic (integrative medicine) or biopsychosocial approach than the more specific bioscientific pathways typical for a pathologically-defined disease. A person-centered approach to evaluation and care more effectively addresses and encompasses the biopsychosocial aspects of this disorder than traditional bioscientific clinical methods. This review outlines a holistic multi-modal, patientcentered approach to evaluation and care as a framework for primary clinic settings. Future directions in research, diagnosis, and management of fibromyalgia patients should incorporate revised person-centered and other qualitative models of care for critical comparison to current conventional concepts and clinical practice. The more comprehensive personcentered services need to be compared to the current standardized practice in terms of their cost-effective outcomes, patient satisfaction, physician gratification, and practical logistics of providing long-term follow up and management.


Changes in the microbial communities associated with Gorgonia ventalina during aspergillosis infection.
The surface mucopolysaccharide layer (SML) secreted by corals is a rich environment where bacteria live and proliferate, with population levels often being several orders of magnitude higher than in the surrounding waters (at least for culturable microbes). Some studies have suggested that these communities play an important role in energy and nutrient flux in marine environments. We hypothesize that the microbial community structure of the SML also plays a role in protection against disease. This hypothesis is based on studies that have shown differences in the bacterial composition of the mucus of healthy and diseased corals. In this study we tested the differences in the microbial communities living in association with the SML of healthy and diseased Gorgonia ventalina by comparing their metabolic profiles using Biolog EcoPlates. Overall, metabolic profiles of the coral surface microbiota were significantly different to those in the water column based on stepwise canonical discriminant analyses (CDAs). Furthermore, differences between communities living in healthy and diseased corals were also found. Changes were observed between affected and unaffected areas of the same colony, although these changes were not as obvious as between individual healthy and diseased colonies. Results suggest that the microbial communities living in the SML of G. ventalina are affected by the presence of aspergillosis, even if the area is not in direct contact with the infection. This suggests the possibility of changes in the composition of the SML throughout the colony as a response to aspergillosis infection.


Alpha waves: a neural signature of visual suppression.
Alpha waves are traditionally considered a passive consequence of the lack of stimulation of sensory areas. However, recent results have challenged this view by showing a modulation of alpha activity in cortical areas representing unattended information during active tasks. These data have led us to think that alpha waves would support a 'gating function' on sensorial stimulation that actively inhibits unattended information in attentional tasks. Visual suppression occurring during a saccade and blink entails an inhibition of incoming visual information, and it seems to occur at an early processing stage. In this study, we hypothesized that the neural mechanism through which the visual system exerts this inhibition is the active imposition of alpha oscillations in the occipital cortex, which in turn predicts an increment of alpha amplitude during a visual suppression phenomena. We measured visual suppression occurring during short closures of the eyelids, a situation well suited for EEG recordings and stimulated the retinae with an intra-oral light administered through the palate. In the behavioral experiment, detection thresholds were measured with eyes steady open and steady closed, showing a reduction of sensitivity in the latter case. In the EEG recordings performed under identical conditions we found stronger alpha activity with closed eyes. Since the stimulation does not depend on whether the eyes were open or closed, we reasoned that this should be a central effect, probably due to a functional role of alpha oscillation in agreement with the 'gating function' theory.


Antigenic specificity of neutralizing antibody to cholera toxin.
Selected rabbit antisera to cholera toxin antigens and convalescent cholera patient sera were analyzed using the permeability factor neutralization test and two sensitive in vitro serological assays specific for cholera toxin, cholera toxin A subunit, and cholera toxin B subunit. The results indicated that antisera to cholera toxin contained toxin-neutralizing activity as well as antibodies specific for both the A subunit and B subunit. It was clearly established that antisera to B subunit, devoid of significant anti-A subunit activity, neutralized the vascular permeability activity of cholera toxin. Antisera to A subunit contained neutralizing antibodies and antibodies to both A and B subunits. Absorption with B subunit removed both the toxin-neutralizing and anti-B subunit activities, while the anti-A activity was unaffected. Neutralizing antibody titers of rabbits immunized with B subunit were also observed to be significantly higher than neutralizing antibody titers of sera from A subunit-immunized rabbits, despite the overall similarity in anti-B subunit titers as determined by passive hemagglutination and radioimmunoassay of sera from the two groups of rabbits. Anti-alpha chain sera neither neutralized cholera toxin nor possessed significant antitoxin or anti-B subunit titers as determined by passive hemagglutination and radioimmunoassay. The anti-alpha chain sera contained high levels of antibody specific for A subunit, which is consistent with the hypothesis that the alpha chain is part of the A subunit structure. In contrast, the gamma chain was not shown to be antigenic. Sera from convalescent cholera patients possessed toxin-neutralizing antibody as well as passive hemagglutination and radioimmunoassay antibody against both A and B subunits.


Direct evidence that thromboxane mimetic U44069 preferentially constricts the afferent arteriole.
The thromboxane A2 (TXA2) mimetic U44069 has been demonstrated to reduce the GFR and filtration fraction of the normal isolated perfused rat kidney markedly, suggesting a predominant constriction of preglomerular vessels. To assess this possibility directly, effects of U44069 on the renal microvessels of the isolated perfused hydronephrotic kidney were examined. At 10(-6) mol/L, U44069 elicited a 27 +/- 2% decrease in afferent arteriolar (AA) diameter (from 18.8 +/- 0.3 to 13.7 +/- 0.3 micron, P < 0.001). In contrast, efferent arteriolar (EA) diameter decreased by only 9 +/- 1% (from 16.4 +/- 0.5 to 15.0 +/- 0.5 micron, P < 0.001). These effects on both AA and EA were completely reversed by the TXA2 receptor antagonist SQ29548. The calcium antagonist diltiazem reversed U44069-induced AA constriction by 83 +/- 5%. The U44069-induced EA constriction was insensitive to the vasodilator action of diltiazem at concentrations from 10(-8) to 10(-6) mol/L, but at 10(-5) mol/L, diltiazem increased the EA diameter significantly, albeit modestly. Nifedipine also reversed the U44069-induced AA constriction (81 +/- 7%), but failed to inhibit the EA constriction at concentrations from 10(-9) to 10(-6) mol/L. These findings constitute the first direct evidence that a TXA2 agonist preferentially constricts the afferent arteriole. Furthermore, the ability of both the calcium antagonist and SQ29548 to reverse the renal microvascular actions of TXA2 agonists suggests a potential utility of these agents in ameliorating TXA2-induced renal hemodynamic abnormalities.


Cholecystectomy and NAFLD: does gallbladder removal have metabolic consequences?
Pathogenesis of nonalcoholic fatty liver (NAFLD) disease and gallbladder (GB) disease secondary to cholesterol gallstones is complex, yet both conditions share similar associated risk factors, most of them related to the metabolic syndrome. Cholecystectomy, the best treatment for GB disease, is one of the most performed abdominal surgeries worldwide. In this issue of the American Journal of Gastroenterology, Ruhl and Everhart, using data from the Third United States National Health and Nutrition Examination Survey (1988-1994), show that NAFLD is associated with cholecystectomy (odds ratio (OR)=2.4; 1.8-3.3), but not with gallstones (OR=1.1; 0.84-1.4). This finding suggests that cholecystectomy may itself represent a risk factor for NAFLD, which is in line with the recently undisclosed role of the GB and bile acids in systemic metabolic regulation. Thus, cholecystectomy may not be innocuous and may have a major impact on public health by contributing to NAFLD development.


Multimodality treatment for non melanoma skin cancer: a prospective study done on 120 egyptian patients.
Multiple currently accepted treatments for non melanoma skin cancer (NMSC) are now available. To present multimodality treatment for NMSCs and to report any side effects or recurrence during three years follow-up period. This is a prospective study including 120 patients with NMSCs. Patients were subjected to different treatment modalities with follow-up for three years. 103 Patients (85.8%) were treated by surgery with or without post operative radiotherapy, 8 patients (5%) received only radiation therapy, 7 patients (5.8%) treated by shave excision followed by medical treatment (intralesional interferon for 3 months, systemic retinoids and cyclo-oxygenase inhibitor for 6 months), and 2 patients (1.7%) escaped the treatment. All patients treated by shave excision followed by medical treatment showed complete healing of all lesions 6 months after completing the course of treatment. Alternatives to the surgical management of NMSC may be preferred under certain circumstances. BCC - Interferon - Retinoids - SCC.


Characterization of the Fe metalloproteome of a ubiquitous marine heterotroph, Pseudoalteromonas (BB2-AT2): multiple bacterioferritin copies enable significant Fe storage.
Fe is a critical nutrient to the marine biological pump, which is the process that exports photosynthetically fixed carbon in the upper ocean to the deep ocean. Fe limitation controls photosynthetic activity in major regions of the oceans, and the subsequent degradation of exported photosynthetic material is facilitated particularly by marine heterotrophic bacteria. Despite their importance in the carbon cycle and the scarcity of Fe in seawater, the Fe requirements, storage and cytosolic utilization of these marine heterotrophs has been less studied. Here, we characterized the Fe metallome of Pseudoalteromonas (BB2-AT2). We found that with two copies of bacterioferritin (Bfr), Pseudoalteromonas possesses substantial capacity for luxury uptake of Fe. Fe : C in the whole cell metallome was estimated (assuming C : P stoichiometry ∼51 : 1) to be between ∼83 μmol : mol Fe : C, ∼11 fold higher than prior marine bacteria surveys. Under these replete conditions, other major cytosolic Fe-associated proteins were observed including superoxide dismutase (SodA; with other metal SOD isoforms absent under Fe replete conditions) and catalase (KatG) involved in reactive oxygen stress mitigation and aconitase (AcnB), succinate dehydrogenase (FrdB) and cytochromes (QcrA and Cyt1) involved in respiration. With the aid of singular value decomposition (SVD), we were able to computationally attribute peaks within the metallome to specific metalloprotein contributors. A putative Fe complex TonB transporter associated with the closely related Alteromonas bacterium was found to be abundant within the Pacific Ocean mesopelagic environment. Despite the extreme scarcity of Fe in seawater, the marine heterotroph Pseudoalteromonas has expansive Fe storage capacity and utilization strategies, implying that within detritus and sinking particles environments, there is significant opportunity for Fe acquisition. Together these results imply an evolved dedication of marine Pseudoalteromonas to maintaining an Fe metalloproteome, likely due to its dependence on Fe-based respiratory metabolism.


Health, trust, or "just understood": explicit and implicit condom decision-making processes among black, white, and interracial same-sex male couples.
Among gay and bisexual men, primary partners are a leading source of HIV infection. Trust, intimacy, and advancements in HIV treatment may impact same-sex male (SSM) couples' decisions to engage in unprotected anal intercourse (UAI). This qualitative study explored how Black, White and interracial couples discussed, and made decisions regarding condoms. Qualitative interviews were conducted with 48 SSM couples in the New York and San Francisco metropolitan areas. Stratified purposive sampling was used to include Black (n = 16), White (n = 17), and interracial (Black-White) (n = 15) couples. Twenty-six couples were concordant HIV-negative and 22 were HIV-discordant. Interviews were recorded, transcribed, coded, and analyzed using a grounded theory approach. Some couples described explicit processes, which involved active discussion, while others described implicit processes, where condom-use decisions occurred without any explicit discussion. These processes also differed by race and HIV status. Black couples tended to report condom-use as "just understood." White, HIV-discordant couples decided not to use condoms, with some identifying the HIV-positive partner's suppressed viral load and high CD4 count as deciding factors. After an unplanned episode of UAI, White, HIV-negative couples tended to discontinue condom use while Black HIV-negative couples decided to revert to using condoms. HIV prevention efforts focused on same-sex, male couples must consider the explicit/implicit nature of condom decision-making processes. Understanding differences in these processes and considering relationship dynamics, across race and HIV status, can promote the development of innovative couple-level, HIV prevention interventions.


Alcohol consumption of UK members of parliament: cross-sectional survey.
This study examined the prevalence of risky drinking by members of parliament (MPs), as well as the relationship between risky drinking and age, years spent as an MP, working outside parliament, awareness of the Parliamentary Health and Wellbeing Service, and probable mental ill health. A survey questionnaire assessed alcohol consumption using the Alcohol Use Disorders Identification Test (AUDIT). Risky drinking was identified by combining categories of increasing (hazardous), higher (harmful) and probable dependent drinking for those with a total score of 8 or more. Comparator groups from the 2014 Adult Psychiatric Morbidity Survey (APMS) were used as controls. UK House of Commons. 650 MPs. Compared with all 650 MPs, participants (n=146) were more likely to be female (p<0.05) or have an educational qualification (p<0.05). Weighted proportions on AUDIT items were higher than the APMS comparator group for participants who had a drink four or more times a week, 10 or more drinks on a typical drinking day, six or more drinks in one occasion, or felt guilty because of drinking (p<0.01). Weighted percentages for risky drinking were higher in MPs compared with the whole English population (p<0.05), but similar when compared with socioeconomic comparator groups. The odds of risky drinking were 2.74 times greater for MPs who had an additional work role outside parliament compared with those who did not (95% CI 0.98 to 7.65) and 2.4 times greater for MPs with probable mental ill health compared with those with no evidence of probable mental ill health (95% CI 0.78 to 7.43). A low level of awareness of the Parliamentary Health and Wellbeing Service has implications for improving the detection of risky drinking and improving access to this service by MPs. Possible increased likelihood of risky drinking in MPs who also had an additional work role outside Parliament and among those with probable mental ill health requires further exploration.


Rotavirus vaccines and vaccination in Latin America.
Worldwide, rotaviruses account for more than 125 million cases of infantile gastroenteritis and nearly 1 million deaths per year, mainly in developing countries. Rather than other control measures, vaccination is most likely to have a major impact on rotavirus disease incidence. The peak incidence of rotavirus diarrhea occurs between 6 and 24 months of age. In developing countries, however, cases are not uncommon among children younger than 6 months. G serotypes 1 to 4 are responsible for most disease, but there are indications that in Brazil that G type 5 is of emerging epidemiological importance. Both homotypic and heterotypic responses are elicited during natural rotavirus infection, and the immunological response at the intestinal mucosal surface is probably the more consistent predictor of clinical immunity. With the primary objective of protecting children against life-threatening dehydrating diarrhea, many approaches to rotavirus vaccine development have been attempted. One vaccine, the tetravalent rhesus-human reassortant rotavirus vaccine (RRV-TV), was given licensing approval in the United States of America, introduced to the market, and later withdrawn. A number of studies have found better efficacy of RRV-TV in developed countries than in developing ones. Field trials with a 4 x 10(4) plaque-forming units (PFU) preparation of RRV-TV have been carried out in two countries in Latin America, Brazil and Peru. Those trials yielded protective efficacy rates against all rotavirus diarrhea ranging from 18% to 35%. Data from a large catchment trial in Venezuela with a higher RRV-TV dose, of 4 x 10(5) PFU/dose, indicated an efficacy rate of 48% against all rotavirus diarrhea and 88% against severe rotavirus diarrhea. It appears that breast-feeding does not compromise the efficacy of RRV-TV if three doses of the vaccine are administered. Similarly, possible interference of oral poliovirus vaccine with the "take" of the rotavirus vaccine can be overcome by giving three doses of the rotavirus vaccine or by using a higher-titer formulation of it. Wild enteroviruses, however, may cause primary rotavirus vaccine failure in developing countries. Studies in Peru with RRV-TV have shown a trend towards higher vaccine efficacy rates against "pure" (rotavirus-only) diarrheal episodes. Economic analyses made in the United States indicate that a vaccine that costs less than US$ 9 per dose would lead to a net savings in medical costs. To date, however, cost-benefit studies have not been done in developing countries. In the future, it is possible that some Latin American countries might adapt their polio production facilities to the preparation of rotavirus vaccines for human use. A year after RRV-TV was licensed for vaccination of infants in the United States, the occurrence of intussusception as an adverse event led to the vaccine's withdrawal from the market. The implications of that action, particularly for Latin America, will be addressed in this article, including the need to explore alternative rotavirus candidate vaccines, particularly through the conduct of parallel clinical trials in both developed and developing countries.


Association of polymorphisms in HCN4 with mood disorders and obsessive compulsive disorder.
Hyperpolarization activated cyclic nucleotide-gated (HCN) potassium channels are implicated in the control of neuronal excitability and are expressed widely in the brain. HCN4 is expressed in brain regions relevant to mood and anxiety disorders including specific thalamic nuclei, the basolateral amygdala, and the midbrain dopamine system. We therefore examined the association of HCN4 with a group of mood and anxiety disorders. We genotyped nine tag SNPs in the HCN4 gene using Sequenom iPLEX Gold technology in 285 Caucasian patients with DSM-IV mood disorders and/or obsessive compulsive disorder and 384 Caucasian controls. HCN4 polymorphisms were analyzed using single marker and haplotype-based association methods. Three SNPs showed nominal association in our population (rs12905211, rs3859014, rs498005). SNP rs12905211 maintained significance after Bonferroni correction, with allele T and haplotype CTC overrepresented in cases. These findings suggest HCN4 as a genetic susceptibility factor for mood and anxiety disorders; however, these results will require replication using a larger sample.


Evaluation of Deep Learning Strategies for Nucleus Segmentation in Fluorescence Images.
Identifying nuclei is often a critical first step in analyzing microscopy images of cells and classical image processing algorithms are most commonly used for this task. Recent developments in deep learning can yield superior accuracy, but typical evaluation metrics for nucleus segmentation do not satisfactorily capture error modes that are relevant in cellular images. We present an evaluation framework to measure accuracy, types of errors, and computational efficiency; and use it to compare deep learning strategies and classical approaches. We publicly release a set of 23,165 manually annotated nuclei and source code to reproduce experiments and run the proposed evaluation methodology. Our evaluation framework shows that deep learning improves accuracy and can reduce the number of biologically relevant errors by half. © 2019 The Authors. Cytometry Part A published by Wiley Periodicals, Inc. on behalf of International Society for Advancement of Cytometry.


Clonal Spread of Candida glabrata Bloodstream Isolates and Fluconazole Resistance Affected by Prolonged Exposure: a 12-Year Single-Center Study in Belgium.
Candida glabrata is a major cause of candidemia in immunocompromised patients and is characterized by a high-level of fluconazole resistance. In the present study, the acquisition of antifungal resistance and potential clonal spread of C. glabrata were explored at a single center over a 12-year period by analyzing 187 independent clinical C. glabrata bloodstream isolates. One strain was found to be micafungin resistant due to a mutation in the FKS2 gene. Fluconazole resistance remained stable throughout the period and was observed in 20 (10.7%) of the isolates. An analysis of the antifungal consumption data revealed that recent prior exposure to fluconazole increased the risk to be infected by a resistant strain. In particular, the duration of the treatment was significantly longer for patients infected by a resistant isolate, while the total and mean daily doses received did not impact the acquisition of resistance in C. glabrata No link between genotype and resistance was found. However, multilocus variable-number tandem-repeat analyses indicated a potential intrahospital spread of some isolates between patients. These isolates shared the same genetic profiles, and infected patients were hospitalized in the same unit during an overlapping period. Finally, quantitative real-time PCR analyses showed that, unlike that for other ABC efflux pumps, the expression of CgCDR1 was significantly greater in resistant strains, suggesting that it would be more involved in fluconazole (FLC) resistance. Our study provides additional evidence that the proper administration of fluconazole is required to limit resistance and that strict hand hygiene is necessary to avoid the possible spreading of C. glabrata isolates between patients.


Focussing over the edge: adaptive subsurface laser fabrication up to the sample face.
Direct laser writing is widely used for fabrication of subsurface, three dimensional structures in transparent media. However, the accessible volume is limited by distortion of the focussed beam at the sample edge. We determine the aberrated focal intensity distribution for light focused close to the edge of the substrate. Aberrations are modelled by dividing the pupil into two regions, each corresponding to light passing through the top and side facets. Aberration correction is demonstrated experimentally using a liquid crystal spatial light modulator for femtosecond microfabrication in fused silica. This technique allows controlled subsurface fabrication right up to the edge of the substrate. This can benefit a wide range of applications using direct laser writing, including the manufacture of waveguides and photonic crystals.


A prospective randomised phase II trial of thalidomide with carboplatin compared with carboplatin alone as a first-line therapy in women with ovarian cancer, with evaluation of potential surrogate markers of angiogenesis.
To compare the safety and efficacy of thalidomide in combination with carboplatin to carboplatin alone as a first-line therapy in women with ovarian cancer and to evaluate the anti-angiogenic effects of thalidomide by measurement of surrogate markers of angiogenesis. Forty patients with Stage IC-IV ovarian cancer were randomly assigned to receive either carboplatin (AUC 7) intravenously every four weeks for up to six doses (n = 20) or carboplatin at the same dose and schedule, plus thalidomide 100 mg orally daily for six months (n = 20). After median follow-up of 1.95 years, there was no difference in the overall response rate (90% in carboplatin arm, 75% in combination arm; p = 0.41). Increased incidence of symptoms of constipation, dizziness, tiredness and peripheral neuropathy was observed in the combination arm. There was a significant fall in CA-125 and E-selectin in both arms after treatment and VCAM-1 in the carboplatin arm. No significant difference between the two arms was observed in any of the markers analysed. In our trial the addition of thalidomide to carboplatin was well tolerated with no increased efficacy. The fall in some of the angiogenic markers in both groups may reflect tumour response rather than any specific anti-angiogenic effect of thalidomide.


Electrophysiology on Channel-Forming Proteins in Artificial Lipid Bilayers: Next-Generation Instrumentation for Multiple Recordings in Parallel.
Artificial lipid bilayers have been used for several decades to study channel-forming pores and ion channels in membranes. Until recently, the classical two-chamber setups have been primarily used for studying the biophysical properties of pore forming proteins. Within the last 10 years, instruments for automated lipid bilayer measurements have been developed and are now commercially available. This chapter focuses on protein purification and reconstitution of channel-forming proteins into lipid bilayers using a classic setup and on the commercially available systems, the Orbit mini and Orbit 16.


Herpes B virus utilizes human nectin-1 but not HVEM or PILRα for cell-cell fusion and virus entry.
To investigate the requirements of herpesvirus entry and fusion, the four homologous glycoproteins necessary for herpes simplex virus (HSV) fusion were cloned from herpes B virus (BV) (or macacine herpesvirus 1, previously known as cercopithecine herpesvirus 1) and cercopithecine herpesvirus 2 (CeHV-2), both related simian simplexviruses belonging to the alphaherpesvirus subfamily. Western blots and cell-based enzyme-linked immunosorbent assay (ELISA) showed that glycoproteins gB, gD, and gH/gL were expressed in whole-cell lysates and on the cell surface. Cell-cell fusion assays indicated that nectin-1, an HSV-1 gD receptor, mediated fusion of cells expressing glycoproteins from both BV and CeHV-2. However, herpesvirus entry mediator (HVEM), another HSV-1 gD receptor, did not facilitate BV- and CeHV-2-induced cell-cell fusion. Paired immunoglobulin-like type 2 receptor alpha (PILRα), an HSV-1 gB fusion receptor, did not mediate fusion of cells expressing glycoproteins from either simian virus. Productive infection with BV was possible only with nectin-1-expressing cells, indicating that nectin-1 mediated entry while HVEM and PILRα did not function as entry receptors. These results indicate that these alphaherpesviruses have differing preferences for entry receptors. The usage of the HSV-1 gD receptor nectin-1 may explain interspecies transfer of the viruses, and altered receptor usage may result in altered virulence, tropism, or pathogenesis in the new host. A heterotypic cell fusion assay resulting in productive fusion may provide insight into interactions that occur to trigger fusion. These findings may be of therapeutic significance for control of deadly BV infections.


[Hemorheological alterations in hypertensive patients].
The aim of this study was to investigate the blood viscosity profile and to evaluate the influence of plasmatic (fibrinogen) and cellular (erythrocyte aggregation) factors in a group of hypertensive patients, compared with a normotensive group. We worked with anticoagulated blood of both non diabetic hypertensive patients (n=31), and healthy individuals (n=40). The plasmatic viscosity and whole blood determination were obtained with a cone-plate viscometer. Erythrocyte aggregation was studied by microscopical observation and quantified by an Aggregate Shape Parameter (ASP), defined as the relation projected area/perimeter. Fibrinogen was determined by the Clauss method with a coagulometer. A comparison between these groups led us to assert that whole blood viscosity was significantly higher in hypertensive patients than in the controls at all shear rates. Plasma viscosity values only showed significant differences between both groups at low shear rate (1.15 a 11.56 seg(-1)). The hypertensive patients showed irregular and amorphous aggregates so that ASP appeared significantly higher (p< 0.001) in patients with hypertension (0.69 +/- 0.11) than in healthy subjects (0.25 +/- 0.12). Fibrinogen appeared slightly higher (p<0.01) in the hypertensive group than in the normal group. Several hemorheological parameters play important roles in the pathogenesis of hypertension. Among these factors, several hemorheological parameters could be altered in hypertension (hematocrit, plasma fibrinogen level, erythrocyte deformability and aggregability, plasma and whole blood viscosity). An increased RBC aggregation has been identified as an important factor responsible for disturbing blood rheological behavior in the microcirculation. The present study demonstrates an abnormal erythrocyte aggregation, which was detected by increased ASP values that could be responsible for vascular complications in hypertension.


Schmallenberg virus experimental infection of sheep.
Since late 2011, a novel orthobunyavirus, named Schmallenberg virus (SBV), has been implicated in many cases of severely malformed bovine and ovine offspring in Europe. In adult cattle, SBV is known to cause a mild transient disease; clinical signs include short febrile episodes, decreased milk production and diarrhoea for a few days. However, the knowledge about clinical signs and pathogenesis in adult sheep is limited. In the present study, adult sheep of European domestic breeds were inoculated with SBV either as cell culture grown virus or as virus with no history of passage in cell cultures. Various experimental set-ups were used. Sampling included blood collection at different time points during the experimental period and selected organ material at autopsy. Data from this study showed, that the RNAemic period in sheep was as short as reported for cattle; viral genome was detectable for about 3-5 days by real-time RT-PCR. In total, 13 out of 30 inoculated sheep became RNAemic, with the highest viral load in animals inoculated with virus from low cell culture passaged or the animal passaged material. Contact animals remained negative throughout the study. One RNAemic sheep showed diarrhoea for several days, but fever was not recorded in any of the animals. Antibodies were first detectable 10-14 days post inoculation. Viral RNA was detectable in spleen and lymph nodes up to day 44 post inoculation. In conclusion, as described for cattle, SBV-infection in adult sheep predominantly results in subclinical infection, transient RNAemia and a specific antibody response. Maintenance of viral RNA in the lymphoreticular system is observed for an extended period.


Nucleotide excision repair in chronic neurodegenerative diseases.
Impaired DNA repair involving the nucleotide excision repair (NER)/transcription-coupled repair (TCR) pathway cause human pathologies associated with severe neurological symptoms. These clinical observations suggest that defective NER/TCR might also play a critical role in chronic neurodegenerative disorders (ND), such as Alzheimer's and Parkinson's disease. Involvement of NER/TCR in these disorders is also substantiated by the evidence that aging constitutes the principal risk factor for chronic ND and that this DNA repair mechanism is very relevant for the aging process itself. Our understanding of the exact role of NER/TCR in chronic ND, however, is extremely rudimentary; while there is no doubt that defective NER/TCR can lead to neuronal death, evidence for its participation in the etiopathogenesis of ND is inconclusive thus far. Here we summarize the experimental observations supporting a role for NER/TCR in chronic ND and suggest questions and lines of investigation that might help in addressing this important issue. We also present a preliminary yet unprecedented meta-analysis on human brain microarray data to understand the expression levels of the various NER factors in the anatomical areas relevant for chronic ND pathogenesis. In summary, this review intends to highlight elements supporting a role of NER/TCR in these devastating disorders and to propose potential strategies of investigation.


Utility of the Ion S5™ and MiSeq FGx™ sequencing platforms to characterize challenging human remains.
Often in missing persons' and mass disaster cases, the samples remaining for analysis are hard tissues such as bones, teeth, nails, and hair. These remains may have been exposed to harsh environmental conditions, which pose challenges for downstream genotyping. Short tandem repeat analysis (STR) via capillary electrophoresis (CE) is still the gold standard for DNA typing; however, a newer technology known as massively parallel sequencing (MPS) could improve upon our current techniques by typing different and more markers in a single analysis, and consequently improving the power of discrimination. In this study, bone and tooth samples exposed to a variety of DNA insults (cremation, embalming, decomposition, thermal degradation, and fire) were assessed and sequenced using the Precision ID chemistry and a custom AmpliSeq™ STR and iiSNP panel on the Ion S5™ System, and the ForenSeq DNA Signature Prep Kit on the MiSeq FGx™ system, as well as the GlobalFiler™ PCR Amplification Kit on the 3500™ Genetic Analyzer. The results demonstrated that using traditional CE-based genotyping performed as expected, producing a partial or full DNA profile for all samples, and that both sequencing chemistries and platforms were able to recover sufficient STR and SNP information from a majority of the same challenging samples. Run metrics including profile completeness and mean read depth produced good results with each system, considering the degree of damage of some samples. Most sample insults (except decomposed) produced similar numbers of alleles for both MPS systems. Comparable markers produced full concordance between the two platforms.


Crossover sexual offenses.
Crossover sexual offenses are defined as those in which victims are from multiple age, gender, and relationship categories. This study investigates admissions of crossover sexual offending from sex offenders participating in treatment who received polygraph testing. For 223 incarcerated and 266 paroled sexual offenders, sexual offenses were recorded from criminal history records and admissions during treatment coupled with polygraph testing. The majority of incarcerated offenders admitted to sexually assaulting both children and adults from multiple relationship types. In addition, there was a substantial increase in offenders admitting to sexually assaulting victims from both genders. In a group of incarcerated offenders who sexually assaulted children, the majority of offenders admitted to sexually assaulting both relatives and nonrelatives, and there was a substantial increase in the offenders admitting to assaulting both male and female children. Although similar trends were observed for the sample of parolees, the rates were far less dramatic. Parolees appeared to have greater levels of denial, had participated in fewer treatment sessions, and perceived greater supervision restrictions as a result of admitting additional offenses. These findings support previous research indicating that many sexual offenders do not exclusively offend against a preferred victim type.


A metabolically stable analog of 1,4,5-inositol trisphosphate activates a novel K+ conductance in pyramidal cells of the rat hippocampal slice.
IP(s)3, a metabolically stable analog of 1,4,5-inositol trisphosphate (IP3), inhibited action potential firing when injected into hippocampal pyramidal cells. This effect was associated with decreased input resistance, a more negative resting potential, outward rectification at depolarized potentials, and an afterhyperpolarization. The response to IP(s)3 was unaffected by antagonists of Na+, Ca2+, and Cl- conductances, but was sensitive to changes in extracellular K+ concentration. The IP(s)3-induced conductance was voltage-dependent, was activated in 10 ms with depolarization, and was blocked by extracellular Ba2+ or intracellular Ca2+ chelation. It was not suppressed by other K+ conductance antagonists. Thus, IP(s)3 may activate a novel K+ conductance in CA1 pyramidal cells. IP3 itself did not elicit this conductance, suggesting it may be rapidly metabolized in these cells.


The application of DNA and RNA G-quadruplexes to therapeutic medicines.
The intriguing structural diversity in folded topologies available to guanine-rich nucleic acid repeat sequences have made four-stranded G-quadruplex structures the focus of both basic and applied research, from cancer biology and novel therapeutics through to nanoelectronics. Distributed widely in the human genome as targets for regulating gene expression and chromosomal maintenance, they offer unique avenues for future cancer drug development. In particular, the recent advances in chemical and structural biology have enabled the construction of bespoke selective DNA based aptamers to be used as novel therapeutic agents and access to detailed structural models for structure based drug discovery. In this critical review, we will explore the important underlying characteristics of G-quadruplexes that make them functional, stable, and predictable nanoscaffolds. We will review the current structural database of folding topologies, molecular interfaces and novel interaction surfaces, with a consideration to their future exploitation in drug discovery, molecular biology, supermolecular assembly and aptamer design. In recent years the number of potential applications for G-quadruplex motifs has rapidly grown, so in this review we aim to explore the many future challenges and highlight where possible successes may lie. We will highlight the similarities and differences between DNA and RNA folded G-quadruplexes in terms of stability, distribution, and exploitability as small molecule targets. Finally, we will provide a detailed review of basic G-quadruplex geometry, experimental tools used, and a critical evaluation of the application of high-resolution structural biology and its ability to provide meaningful and valid models for future applications (255 references).


Is estimated bullet trajectory a reliable predictor of severe injury? Case report of a thoraco-abdominal gunshot with a protracted trajectory managed nonoperatively.
Operative management of all gunshot's traumas carries a high rate of unwarranted interventions that are known to cause serious complications. Selective nonoperative management is thus being increasingly practiced which has reduced these avoidable interventions. Physical examination and computed tomography scans are most sensitive in assessing need of laparotomy. Assessment of internal injuries on the basis of an estimated bullet trajectory is often practiced but has seldom been studied. We report a case of conservative management of a thoraco abdominal gun shot patient where an estimated bullet trajectory was indicative of serious injuries. To the best of our knowledge this is the first report of a thoraco abdominal gunshot that, despite of a protracted trajectory, had no sequelae and was thus managed nonoperatively. A 30 year old male patient having height of 180 cm and weight of 70 kg (Body Mass Index 21.6) presented with complaint of a penetrating injury at left side of upper torso. The patient had no symptoms or obvious bleeding and was vitally stable. On examination a 1 cm × 1 cm entry wound at the left 3rd intercostal space in the mid clavicular line was identified. The chest and abdomen were otherwise unremarkable on examination. The chest radiograph displayed clear lung fields. The abdominal radiographs displayed a bullet in the upper left quadrant of the abdomen lateral to the spine. The bullets estimated trajectory from 3rd intercostal space and its lodgment in the abdomen lateral to the spine indicated severe visceral injury. The computed tomography scan showed that the bullet was lodged postero-medially to the left kidney. All thoracic, intra peritoneal and retroperitoneal visceral structures were identified to be normal. The patient remained clinically and vitally stable, hence was managed nonoperatively being discharged after 48 h of observation. From this case we conclude that decision for managing gun shot patients should be based on objective clinical and diagnostic findings. We recommend further investigation of the predictability of estimated trajectory for visceral injuries and consequent operative intervention as we found it to be misleading in this case.


Shikonin production by extractive cultivation in transformed-suspension and hairy root cultures of Lithospermum erythrorhizon.
Effects of in situ extraction, fungal elicitation, a permeabilizing agent, and the oxygen transfer rate on shikonin production in transformed suspension and hairy root cultures of Lithospermum erythrorhizon were studied. Shikonin production with in situ extraction in transformed cell and hairy root cultures by n-hexadecane was 7.6 and 3 times higher than those of the control culture. Shikonin production of transformed L. erythrohizon increased with the enhanced gas exchange, and in situ extraction also increased sucrose consumption and shikonin production. The optimal volume of n-hexadecane in the hairy root culture was similar to that in the transformed cell cultures. In situ extraction at an earlier stage significantly enhanced shikonin production both in transformed cell and hairy root cultures. Dimethylsulfoxide used as a permeabilizing agent was harmful to cell growth and shikonin production, and permeabilizing was unnecessary when in situ extraction was applied. This occurred because with the solvent addition, most shikonin was spontaneously released from the cells and was dissolved in the solvent layer. The combined addition of n-hexadecane of the extract of the fungus Penicillium as an elicitor seemed to result in a higher production of shikonin both in cell suspensions and transformed root cultures. However, an increase of shikonin induction by fungal elicitation in a hairy root culture was not significant in comparison with that of normal cell cultures.


Lead contamination of subarctic lakes and its response to reduced atmospheric fallout: can the recovery process be counteracted by the ongoing climate change?
Can a climate-triggered export of old contaminants from the soil alter the lead (Pb) contaminant burden of subarctic lakes? To address this question, we reconstructed the pollution history of three high latitude lakes situated in a region where a recent climatic shift has occurred. Dated sediment records were used as archives of past Pb inputs to the lakes, where the difference in the (206)Pb/(207)Pb ratio between atmospheric contaminants ((206)Pb/(207)Pb ratio <1.16) and geogenic Pb in the catchment soil ((206)Pb/(207)Pb ratio >1.22) were used to trace fluxes of Pb contaminants. Lead contaminants were found in sediments deposited since Roman times. A significant export of Pb from the soil contaminant pool is indicated in two of the lakes surrounded by near-shore permafrost soils. Here, levels of Pb contaminants and (206)Pb/(207)Pb ratios of sediments deposited after the 1970s appear not to have been strongly affected by the >or=90% reduction in atmospheric deposition rates and increasing (206)Pb/(207)Pb ratios of atmospheric Pb since the 1990s. We concluded that soil processes stimulated by the ongoing climate change at high latitudes might work counteractive to efforts to reduce contaminant levels in subarctic lakes.


Accurate, robust, and automated longitudinal and cross-sectional brain change analysis.
Quantitative measurement of brain size, shape, and temporal change (for example, in order to estimate atrophy) is increasingly important in biomedical image analysis applications. New methods of structural analysis attempt to improve robustness, accuracy, and extent of automation. A fully automated method of longitudinal (temporal change) analysis, SIENA, was presented previously. In this paper, improvements to this method are described, and also an extension of SIENA to a new method for cross-sectional (single time point) analysis. The methods are fully automated, robust, and accurate: 0.15% brain volume change error (longitudinal): 0.5-1% brain volume accuracy for single-time point (cross-sectional). A particular advantage is the relative insensitivity to differences in scanning parameters. The methods provide easy manual review of their output by the automatic production of summary images which show the results of the brain extraction, registration, tissue segmentation, and final atrophy estimation.


High-power, fiber-laser-pumped, picosecond optical parametric oscillator based on MgO:sPPLT.
We report a stable, high-power, mid-infrared synchronously-pumped optical parametric oscillator (SPOPO) based on MgO:sPPLT, pumped by a 1064 nm, picosecond Yb-fiber laser operating at a repetition rate of 81.1 MHz. The singly resonant SPOPO is tunable over 1531-1642 nm (111 nm) in the near-infrared signal and 3022-3488 nm (466 nm) in the mid-infrared idler, providing a total tuning range of 577 nm. Careful optimization of output coupling results in a signal output power as high as 4.3 W at 1593 nm and a mid-infrared idler power of 2 W at 3204 nm for 13.4 W of pump power at a total extraction efficiency of 47%. The SPOPO can be operated near room temperature, down to 30 °C, and exhibits passive peak-to-peak power stability better than 8.6% at 1568 nm (signal) and 8.2% at 3310 nm (idler) over 13 hours at full power. The output signal pulses have duration of 17.5 ps, with a FWHM spectral bandwidth of 1.4 nm centered at 1568 nm.


Free-breathing abdominal MRI improved by repeated k-t-subsampling and artifact-minimization (ReKAM).
We report an approach, termed Repeated k-t-subsampling and artifact-minimization (ReKAM), for removing motion artifacts in free-breathing abdominal MRI. The method is particularly valuable for challenging patients who may not hold their breath for a long time or have irregular respiratory rate. The ReKAM framework comprises one acquisition module and two reconstruction modules. A fast MRI sequence is used to repeatedly acquire multiple sets of k-t space data. Motion artifacts are then minimized by two reconstruction modules: (a) a bootstrapping module in k-t-space is used to identify a low-artifact image; (b) a constrained reconstruction module that integrates projection onto convex set (POCS) and multiplexed sensitivity encoding (MUSE), termed POCSMUSE, is applied to further remove residual artifact. The ReKAM framework is compatible with different pulse sequences, and generally applicable to irregular data sampling patterns in k-space. Free-breathing fast spin-echo MRI data, acquired from healthy volunteers and patients, were used to evaluate the developed ReKAM method. Experimental results show that the ReKAM technique can produce high-quality free-breathing images with the artifact levels comparable to that of breath-holding MRI. The ReKAM framework improves the quality of free-breathing abdominal MRI data, and is compatible with various MRI pulse sequences.


Evidence-Based Checklist to Delay Cardiac Arrest in Brain-Dead Potential Organ Donors: The DONORS Cluster Randomized Clinical Trial.
The effectiveness of goal-directed care to reduce loss of brain-dead potential donors to cardiac arrest is unclear. To evaluate the effectiveness of an evidence-based, goal-directed checklist in the clinical management of brain-dead potential donors in the intensive care unit (ICU). The Donation Network to Optimize Organ Recovery Study (DONORS) was an open-label, parallel-group cluster randomized clinical trial in Brazil. Enrollment and follow-up were conducted from June 20, 2017, to November 30, 2019. Hospital ICUs that reported 10 or more brain deaths in the previous 2 years were included. Consecutive brain-dead potential donors in the ICU aged 14 to 90 years with a condition consistent with brain death after the first clinical examination were enrolled. Participants were randomized to either the intervention group or the control group. The intention-to-treat data analysis was conducted from June 15 to August 30, 2020. Hospital staff in the intervention group were instructed to administer to brain-dead potential donors in the intervention group an evidence-based checklist with 13 clinical goals and 14 corresponding actions to guide care, every 6 hours, from study enrollment to organ retrieval. The control group provided or received usual care. The primary outcome was loss of brain-dead potential donors to cardiac arrest at the individual level. A prespecified sensitivity analysis assessed the effect of adherence to the checklist in the intervention group. Among the 1771 brain-dead potential donors screened in 63 hospitals, 1535 were included. These patients included 673 males (59.2%) and had a median (IQR) age of 51 (36.3-62.0) years. The main cause of brain injury was stroke (877 [57.1%]), followed by trauma (485 [31.6%]). Of the 63 hospitals, 31 (49.2%) were assigned to the intervention group (743 [48.4%] brain-dead potential donors) and 32 (50.8%) to the control group (792 [51.6%] brain-dead potential donors). Seventy potential donors (9.4%) at intervention hospitals and 117 (14.8%) at control hospitals met the primary outcome (risk ratio [RR], 0.70; 95% CI, 0.46-1.08; P = .11). The primary outcome rate was lower in those with adherence higher than 79.0% than in the control group (5.3% vs 14.8%; RR, 0.41; 95% CI, 0.22-0.78; P = .006). This cluster randomized clinical trial was inconclusive in determining whether the overall use of an evidence-based, goal-directed checklist reduced brain-dead potential donor loss to cardiac arrest. The findings suggest that use of such a checklist has limited effectiveness without adherence to the actions recommended in this checklist. ClinicalTrials.gov Identifier: NCT03179020.


Electrochemical Skin Conductance Correlates with Skin Nerve Fiber Density.
Electrochemical skin conductance (ESC) using reverse iontophoresis and chronoamperometry has been used to evaluate abnormal function of small fibers. How ESC correlates with loss of small fibers in skin is unclear. This was a prospective, blinded study. The primary outcome measure was the correlation between ESC at the feet and results of skin biopsies including epidermal nerve fiber density (ENFD) and sweat gland nerve fiber density (SGNFD) at the distal leg. ESC, ENFD, and SGNFD data were normalized by adjusting for weight. The secondary outcome measures were the correlation between ESC and the following variables: quantitative sudomotor axon reflex test (QSART) and symptom scales (neuropathy, pain and autonomic). Eighty-one patients (mean ± sd): age = 53.3 ± 17.3, men/women = 25/56 were enrolled in the study. ESC was reduced in subjects with abnormally low ENFD (ENFD normal/abnormal, ESC = 1.17 ± 0.27/0.87 ± 0.34 μSiemens/kg, p < 0.0008) and abnormally low SGNFD (SGNFD normal/abnormal ESC = 1.09 ± 0.34/0.78 ± 0.3 μSiemens/kg, p < 0.0003). ESC correlated with ENFD (ρ = 0.73, p = 0.0001) and SGNFD (ρ = 0.64, p = 0.0001). ESC did not correlate with symptom scales. ESC is diminished in subjects who have a reduced number of small fibers in the skin and the ESC reduction is proportional to ENFD and SGNFD. ESC can be useful in detecting loss of small nerve fibers.


Ischemic heart disease and ankylosing spondylitis-assessing the role of inflammation.
To assess the association of ankylosing spondylitis (AS) and ischemic heart disease (IHD) compared to traditional cardiovascular (CV) risk factors. Primary care and hospital records of patients with AS were analyzed, using the largest health maintenance organization in Israel, the "Clalit" Health Services data. These patients were compared with age- and gender-matched controls regarding the proportion of IHD in a cross-sectional study. Parameters including socioeconomic status, body mass index (BMI), smoking habits, and coexistent medical conditions hypertension, hyperlipidemia, and diabetes mellitus (DM) - as well as the use of NSAIDs and anti-TNFs were also assessed. The study included 4076 AS patients compared to 20,290 age- and gender-matched controls without AS. The proportion of IHD was higher among AS patients as compared to controls (14.1 vs. 6.36%, respectively, p < 0.01) and patients treated with anti-TNFs had a lower risk for IHD compared to non-anti-TNF users. The proportion of hypertension, hyperlipidemia, DM, and smoking was also higher among AS patients. However, in multivariate analyses following adjustment to these risk factors, AS was not found to be associated with IHD nor anti-TNF therapy to be a protective factor. Patients with AS have more traditional CV risk factors, thus are in a higher risk for IHD. AS itself was not shown to be independently associated with IHD. These findings emphasize the multifactorial process leading to increased proportion of IHD among AS patients and the need for a stringent control of traditional risk factors in these patients.


Present state of metal-on-metal hybrid hip resurfacing.
Bone conservation and preservation, joint stability, and low wear of the large metal-on-metal resurfacing bearings have been convincingly demonstrated in the current literature. The clinical results of 600 MM Hybrid Conserve Plus Resurfacing in 519 patients with an average follow-up of 6.9 years (range, 4.0-10.4 years) have been excellent. The average age was 48.9 years, 74% of the patients were male, and the study included all etiologies of the young with arthritis. The complication rates other than dislocation and fracture of the femoral neck are comparable between resurfacing and conventional total hip replacement. The incidence of femoral neck fracture is low (1.2% worldwide) with less than 0.6% in this series and none occurring in the last 5 years due to proper patient selection and improved surgical technique. Component loosening after metal-on-metal resurfacing has been significantly reduced and acetabular component loosening is uncommon and has not happened in this series. Femoral bone preparation and optimal cementing techniques are paramount to prevention of femoral loosening. Clearance between the cylindrically reamed part of the head and the component varies in different designs, and the surgeon must note the need for different cementing strategies for different recommended clearances. The learning curve of a surgeon undertaking resurfacing can be greatly reduced by observation and hands-on training in specialized centers with surgeons experienced in resurfacing.


Failure of desmopressin to lower serum sodium or prevent crisis in patients with sickle cell anemia.
An analogue of arginine vasopressin (desmopressin, DDAVP) was evaluated for production chronic hyponatremia and prevention of sickle cell crisis. With sodium restriction (100 meq Na + / day) and water loading ( greater than 3 liters/day), persistent hyponatremia could not be achieved, nor could crises be prevented or aborted. Patients would not comply with a regimen of lower salt and higher fluid intake. More rigorous treatment might be practical during acute sickle cell crises, and a regimen similar to that used here might be more effective in children, whose renal concentrating mechanisms are still intact.


Chiropractic chronic low back pain sufferers and self-report assessment methods. Part I. A reliability study of the Visual Analogue Scale, the Pain Drawing and the McGill Pain Questionnaire.
Comprehensive assessment of the problems of low back pain sufferers requires that (in addition to objective measures and clinicians' judgments) an examination of subjective perception of their difficulties should also be made. Subjective measurement has, in the past, presented several problems to researchers. However, in recent years, several new methods of exploring individual reactions to pain and disability have been developed. These assessment measures have, in the main, been carefully explored with severe chronic, acutely ill and surgical patients, but they have not been shown to be appropriate for use with mild chronic low back pain sufferers. This study examines the reliability of several widely used self-report measures. In general, the measures were shown to have acceptable levels of reliability for use with this population, although recommendations for caution in the use of the pain drawing is urged.


Youth Risk Behavior Surveillance: National College Health Risk Behavior Survey--United States, 1995.
Colleges and universities are important settings for delivering health promotion education and services to many young adults. However, before this national college-based survey was conducted in 1995, the prevalences of health-risk behaviors among college students nationwide had not been well characterized. January through June 1995. The Youth Risk Behavior Surveillance System (YRBSS) monitors six categories of priority health-risk behaviors among youth and young adults: behaviors that contribute to unintentional and intentional injuries, tobacco use, alcohol and other drug use, sexual behaviors, unhealthy dietary behaviors, and physical inactivity. The YRBSS includes a) national, state, and local school-based surveys of high school students conducted biennially since 1991, b) a household-based survey conducted in 1992 among a national sample of youth aged 12-21 years, whether enrolled in school, and c) the national college-based survey conducted in 1995. This report summarizes results from the national college-based survey-the National College Health Risk Behavior Survey (NCHRBS)-and describes priority health-risk behaviors among college students nationwide and health promotion programs on college campuses. Data from the 1995 survey indicated that many college students throughout the United States engage in behaviors that place them at risk for serious health problems. Almost one third (29.0%) of college students were current cigarette smokers. One third (34.5%) of college students reported episodic heavy drinking during the 30 days preceding the survey, 27.4% reported drinking alcohol and driving during the 30 days preceding the survey, and 30.5% of students who had gone boating or swimming during the 12 months preceding the survey had drunk alcohol while boating or swimming. One in five (20.4%) female college students had been forced to have sexual intercourse during her lifetime. Only 29.6% of students who had had sexual intercourse during the 3 months preceding the survey had used a condom at last sexual intercourse, and 34.5% had used birth control pills. Approximately one in five (20.5%) college students was overweight. Survey results indicated that three fourths (73.7%) of students had failed to eat five or more servings of fruits and vegetables on the day preceding the survey, 21.8% had eaten three or more high-fat foods on the day preceding the survey, and few students had engaged in vigorous (37.6%) or moderate (19.5%) physical activity at recommended levels. The NCHRBS data will be used to measure progress toward achieving 28 national health objectives related to the health-risk behaviors of college students and two national health objectives related to the availability and characteristics of health promotion programs for college students. These data also will be used nationwide by college health and education officials to improve health policies and programs designed to reduce risks associated with the leading causes of mortality and morbidity among college students.


Pulsed blue laser at 491 nm by nonlinear cavity dumping.
A nonlinear cavity dumping process is applied for the first time to generate kW peak power pulses at 491 nm. The system is based on efficient sum-frequency mixing of 1063 nm and 912 nm radiations in a BiBO nonlinear crystal placed inside a Nd:GdVO4 laser oscillator with a high finesse cavity at 912 nm. The nonlinear cavity dumping process is triggered by high peak power nanosecond pulses from a 1063 nm Q-switched Nd:GdVO4 laser operating at 10 kHz. To reach the kW range at 491 nm a key point is to Q-switch the high finesse 912 nm cavity instead of continuous wave operation. Thus, the peak power (9.3 kW for 3 ns pulses) and the average power (280 mW) obtained at 491 nm are 14 times higher than the one obtained when the 912 nm laser operated in continuous wave.


An efficient and scalable extraction and quantification method for algal derived biofuel.
Microalgae are capable of synthesizing a multitude of compounds including biofuel precursors and other high value products such as omega-3-fatty acids. However, accurate analysis of the specific compounds produced by microalgae is important since slight variations in saturation and carbon chain length can affect the quality, and thus the value, of the end product. We present a method that allows for fast and reliable extraction of lipids and similar compounds from a range of algae, followed by their characterization using gas chromatographic analysis with a focus on biodiesel-relevant compounds. This method determines which range of biologically synthesized compounds is likely responsible for each fatty acid methyl ester (FAME) produced; information that is fundamental for identifying preferred microalgae candidates as a biodiesel source. Traditional methods of analyzing these precursor molecules are time intensive and prone to high degrees of variation between species and experimental conditions. Here we detail a new method which uses microwave energy as a reliable, single-step cell disruption technique to extract lipids from live cultures of microalgae. After extractable lipid characterization (including lipid type (free fatty acids, mono-, di- or tri-acylglycerides) and carbon chain length determination) by GC-FID, the same lipid extracts are transesterified into FAMEs and directly compared to total biodiesel potential by GC-MS. This approach provides insight into the fraction of total FAMEs derived from extractable lipids compared to FAMEs derived from the residual fraction (i.e. membrane bound phospholipids, sterols, etc.). This approach can also indicate which extractable lipid compound, based on chain length and relative abundance, is responsible for each FAME. This method was tested on three species of microalgae; the marine diatom Phaeodactylum tricornutum, the model Chlorophyte Chlamydomonas reinhardtii, and the freshwater green alga Chlorella vulgaris. The method is shown to be robust, highly reproducible, and fast, allowing for multiple samples to be analyzed throughout the time course of culturing, thus providing time-resolved information regarding lipid quantity and quality. Total time from harvesting to obtaining analytical results is less than 2h.


Prevalence of postnatal psychiatric morbidity in mothers and fathers.
In the first study to systematically examine postnatal depression in fathers, we examined depression in 200 postnatal couples, using a two-stage design. The prevalence of depression ascertained by the 13-item Edinburgh Postnatal Depression Scale (EPDS), using a cut-off score for 'caseness' of 13 or more in an unselected postnatal sample, was 27.5% in mothers at six weeks postpartum, 25.7% in mothers at six months postpartum, 9.0% in fathers at six weeks postpartum, and 5.4% in fathers at six months postpartum. The prevalence did not differ significantly in either mothers or fathers from a control group of parents with children between three and five years of age. As expected, mothers had a significantly higher prevalence of psychiatric 'caseness' at both six weeks and six months postpartum than fathers. Fathers were significantly more likely to be cases if their partners were also cases. The hypothesis that different aetiological factors would be important in brief and persistent disorders in mothers was upheld.


Polygenic risk scores for major depressive disorder and neuroticism as predictors of antidepressant response: Meta-analysis of three treatment cohorts.
There are currently no reliable approaches for correctly identifying which patients with major depressive disorder (MDD) will respond well to antidepressant therapy. However, recent genetic advances suggest that Polygenic Risk Scores (PRS) could allow MDD patients to be stratified for antidepressant response. We used PRS for MDD and PRS for neuroticism as putative predictors of antidepressant response within three treatment cohorts: The Genome-based Therapeutic Drugs for Depression (GENDEP) cohort, and 2 sub-cohorts from the Pharmacogenomics Research Network Antidepressant Medication Pharmacogenomics Study PRGN-AMPS (total patient number = 760). Results across cohorts were combined via meta-analysis within a random effects model. Overall, PRS for MDD and neuroticism did not significantly predict antidepressant response but there was a consistent direction of effect, whereby greater genetic loading for both MDD (best MDD result, p < 5*10-5 MDD-PRS at 4 weeks, β = -0.019, S.E = 0.008, p = 0.01) and neuroticism (best neuroticism result, p < 0.1 neuroticism-PRS at 8 weeks, β = -0.017, S.E = 0.008, p = 0.03) were associated with less favourable response. We conclude that the PRS approach may offer some promise for treatment stratification in MDD and should now be assessed within larger clinical cohorts.


Plasma amino acid profiles of breast cancer patients early in the trajectory of the disease differ from healthy comparison groups.
This study describes and compares fasting plasma amino acid profiles of breast cancer patients near the initiation of chemotherapy with those of healthy age- and body mass index-matched females (HM), as well as young healthy females (HY). Breast cancer patients had significantly greater glutamate and histidine concentrations and significantly lower threonine concentrations compared with HM and HY females independent of protein or caloric intake. These differences may be related to metabolic perturbations associated with the disease.


Supplemented ERA-EDTA Registry data evaluated the frequency of dialysis, kidney transplantation, and comprehensive conservative management for patients with kidney failure in Europe.
The aims of this study were to determine the frequency of dialysis and kidney transplantation and to estimate the regularity of comprehensive conservative management (CCM) for patients with kidney failure in Europe. This study uses data from the ERA-EDTA Registry. Additionally, our study included supplemental data from Armenia, Germany, Hungary, Ireland, Kosovo, Luxembourg, Malta, Moldova, Montenegro, Slovenia and additional data from Israel, Italy, Slovakia using other information sources. Through an online survey, responding nephrologists estimated the frequency of CCM (i.e. planned holistic care instead of kidney replacement therapy) in 33 countries. In 2016, the overall incidence of replacement therapy for kidney failure was 132 per million population (pmp), varying from 29 (Ukraine) to 251 pmp (Greece). On 31 December 2016, the overall prevalence of kidney replacement therapy was 985 pmp, ranging from 188 (Ukraine) to 1906 pmp (Portugal). The prevalence of peritoneal dialysis (114 pmp) and home hemodialysis (28 pmp) was highest in Cyprus and Denmark respectively. The kidney transplantation rate was nearly zero in some countries and highest in Spain (64 pmp). In 28 countries with five or more responding nephrologists, the median percentage of candidates for kidney replacement therapy who were offered CCM in 2018 varied between none (Slovakia and Slovenia) and 20% (Finland) whereas the median prevalence of CCM varied between none (Slovenia) and 15% (Hungary). Thus, the substantial differences across Europe in the frequency of kidney replacement therapy and CCM indicate the need for improvement in access to various treatment options for patients with kidney failure.


Cell biology and immunology of Listeria monocytogenes infections: novel insights.
Listeria monocytogenes is an intracellular bacterium responsible for a disease characterized by several clinical features, such as septicemia, brain infection, abortion, and perinatal infection. These clinical features are attributed to its amazing capacity to cross several barriers in susceptible hosts. Intracellular infection is a consequence of the bacterium's capacity to enter a wide variety of mammalian cells, to not only survive but also replicate therein, and to its faculty to spread from one cell to the next, thereby escaping the humoral immune response. Here, we review both the well-established and the newly discovered strategies used by this bacterium to achieve this intracellular lifestyle while escaping from the host innate immune response. More than ever, Listeria appears as a model system and a reference in infection biology.


Should epitope-based HLA compatibility be used in the kidney allocation system?
The new kidney allocation system (KAS) still applies donor-recipient HLA compatibility mostly at the antigen level and although some four-digit alleles have been included. This system is used to record unacceptable mismatches for sensitized transplant candidates with serum HLA antibodies. Since the reactivities of such antibodies are specifically associated with epitopes rather than HLA antigens, a more scientifically accurate assessment of mismatch acceptability could be based on epitopes. HLA class I and class II epitope specificity analyses can now be readily performed with serum antibody assays with single allele panels. This report describes an epitope-based HLA compatibility system for KAS and involves recipient and donor HLA typing at the four-digit allele level. It focuses on sensitized patients who have serum antibodies specific for HLA epitopes that can be entered as unacceptable mismatches in the transplant candidate database. Newly developed software programs could readily identify compatible HLA types.


How much does reducing inequality matter for global poverty?
The goals of ending extreme poverty by 2030 and working towards a more equal distribution of incomes are part of the United Nations' Sustainable Development Goals. Using data from 166 countries comprising 97.5% of the world's population, we simulate scenarios for global poverty from 2019 to 2030 under various assumptions about growth and inequality. We use different assumptions about growth incidence curves to model changes in inequality, and rely on a machine-learning algorithm called model-based recursive partitioning to model how growth in GDP is passed through to growth as observed in household surveys. When holding within-country inequality unchanged and letting GDP per capita grow according to World Bank forecasts and historically observed growth rates, our simulations suggest that the number of extreme poor (living on less than $1.90/day) will remain above 600 million in 2030, resulting in a global extreme poverty rate of 7.4%. If the Gini index in each country decreases by 1% per year, the global poverty rate could reduce to around 6.3% in 2030, equivalent to 89 million fewer people living in extreme poverty. Reducing each country's Gini index by 1% per year has a larger impact on global poverty than increasing each country's annual growth 1 percentage point above forecasts. We also study the impact of COVID-19 on poverty and find that the pandemic may have driven around 60 million people into extreme poverty in 2020. If the pandemic increased the Gini index by 2% in all countries, then more than 90 million may have been driven into extreme poverty in 2020.


Evaluation of Plasma miR-21 and miR-152 as Diagnostic Biomarkers for Common Types of Human Cancers.
Stable blood based miRNA species have allowed for the differentiation of patients with various types of cancer. Therefore, specific blood-based miRNA might be considered as a methodology which could be informative of the presence of cancer potentially from multiple distinct organ sites. Recently, miR-21 has been identified as an "oncomir" in various tumors while miR-152 as a tumor suppressor. In this study, we investigated whether circulating miR-21 and miR-152 can be used for early detection of lung cancer (LuCa), colorectal carcinoma (CRC), breast cancer (BrCa) and prostate cancer (PCa), with distinguishing cancer from various benign lesions on these organ sites. We measured the two miRNA levels by using real-time RT-PCR in plasma samples from a total of 204 cancer patients, 159 various benign lesions, and 228 normal subjects. We observed significantly elevated expression of miR-21 and miR-152 in LuCa, CRC, and BrCa when compared with normal controls. We also found upregulation of plasma miR-21 and miR-152 levels in patients with benign lesions of lung and breast, as compared to normal controls, respectively. No significant expression variation of the two miRNAs was observed in PCa or prostatic benign lesions as compared to healthy controls. Receiver operating characteristic (ROC) analyses revealed that miR-21 and/or miR-152 can discriminate LuCa, CRC and BrCa from normal controls. Our results suggest that plasma miR-21 and miR-152 may serve as non-specific noninvasive biomarkers for early screening of LuCa, CRC, and BrCa, but not PCa.


Cordycepin, a Natural Antineoplastic Agent, Induces Apoptosis of Breast Cancer Cells via Caspase-dependent Pathways.
Cordycepin, a major compound separated from Cordyceps sinensis, is known as a potential novel candidate for cancer therapy. Breast cancer, the most typical cancer diagnosed among women, remains a global health problem. In this study, the anti-breast cancer property of cordycepin and its underlying mechanisms was investigated. The direct effects of cordycepin on breast cancer cells both in in vitro and in vivo experiments were evaluated. Cordycepin exerted cytotoxicity in MCF-7 and MDA-MB-231 cells confirmed by reduced cell viability, inhibition of cell proliferation, enhanced lactate dehydrogenase release and reactive oxygen species accumulation, induced mitochondrial dysfunction and nuclear apoptosis in human breast cancer cells. Cordycepin increased the activation of pro-apoptotic proteins, including caspase-8, caspase-9, caspase-3 and Bax, and suppressed the expression of the anti-apoptotic protein, B-cell lymphoma 2 (Bcl-2). The inhibition on MCF-7-xenografted tumor growth in nude mice further confirmed cordycepin's anti-breast cancer effect. These aforementioned results reveal that cordycepin induces apoptosis in human breast cancer cells via caspase-dependent pathways. The data shed light on the possibility of cordycepin being a safe agent for breast cancer treatment.


Bound volatile precursors in genotypes in the pedigree of 'Marion' blackberry (Rubus sp.).
Glycosidically bound volatiles and precursors in genotypes representing the pedigree for 'Marion' blackberry were investigated over two growing seasons. The volatile precursors were isolated using a C18 solid-phase extraction column. After enzymatic hydrolysis, the released volatiles were analyzed using stir bar sorptive extraction gas chromatography-mass spectrometry (GC-MS) and direct microvial insert thermal desorption GC-MS. The most abundant volatile precursors in the genotypes were alcohols, followed by shikimic acid derivatives. High amounts of furanone glycosides were also detected, while norisoprenoids only existed in a small amount in blackberries. The volatile precursor composition in the genotypes in the 'Marion' pedigree was very similar to their free volatile distribution. 'Logan' and 'Olallie' predominantly had bound norisoprenoids. Wild 'Himalaya' predominated with terpene alcohol and furaneol glycosides, whereas 'Santiam' and 'Chehalem' contained a high level of terpene alcohol glycosides. A similar inheritance pattern was also observed for some volatile precursors in the genotypes in the 'Marion' pedigree. A high content of linalool, hydroxylinalool, and alpha-ionol glycosides in 'Olallie' and a low content in 'Chehalem' resulted in a moderate level in their offspring 'Marion', while a low content of (E)-linalool oxide precursor in 'Olallie' and a high content in 'Chehalem' also resulted in a moderate level in 'Marion'. However, the concentration of furaneol glycosides in 'Marion' exceeded that of its two parents.


How a grafting anchor tailors the cellular uptake and in vivo fate of dendronized iron oxide nanoparticles.
Superparamagnetic spherical iron oxide nanoparticles of 10 nm diameter have been synthesized by thermal decomposition and grafted through a direct ligand exchange protocol with two dendrons bearing respectively a monophosphonic anchor (D2) or a biphosphonic tweezer (D2-2P) at their focal point. Physico-chemical characterization techniques such as dynamic light scattering (DLS), zeta potential, Fourier transform infrared spectroscopy (FTIR), transmission electron microscopy (TEM) and superconducting quantum interference device (SQUID) magnetometry were used to assess their composition, colloidal stability and magnetic properties. High-resolution magic angle spinning (HR-MAS) nuclear magnetic resonance (NMR) spectroscopy studies have been conducted to understand the organic shell composition and to determine both the grafting rate of the dendrons onto the nanoparticle surface and the influence of the remaining oleic acid originating from the synthesis protocol on the cellular uptake. Both dendronized IONPs showed moderate in vitro toxicity (MTT and LDH tests) in human cancer and primary cell lines. Furthermore, in vivo MRI studies showed high contrast enhancement as well as renal and hepatobiliary excretions and highlighted the influence of the grafting anchor (mono- versus bi-phosphonate) on the in vivo fate of dendronized magnetic iron oxides.


Characteristics of Early Paget's Disease in SQSTM1 Mutation Carriers: Baseline Analysis of the ZiPP Study Cohort.
Mutations in SQSTM1 are strongly associated with Paget's disease of bone (PDB), but little is known about the clinical characteristics of those with early disease. Radionuclide bone scans, biochemical markers of bone turnover, and clinical characteristics were analyzed in SQSTM1 mutation carriers who took part in the Zoledronic acid in the Prevention of Paget's disease (ZiPP) study. We studied 222 individuals, of whom 54.9% were female, with mean ± SE age of 50.1 ± 0.6 years. Twelve SQSTM1 mutations were observed, including p.Pro392Leu, which was present in 141 of 222 (63.5%) subjects. Bone scan examination revealed evidence of PDB in 20 subjects (9.0%), ten of whom (50%) had a single affected site. Participants with lesions were older than those without lesions but the difference was not significant (53.6 ± 9.1 versus 49.8 ± 8.9; p = .07). The mean age of participants with lesions was not significantly different from the age at which their parents were diagnosed with PDB (55 years versus 59 years, p = .17). All individuals with lesions were asymptomatic. Serum concentrations of total alkaline phosphatase (ALP) normalized to the upper limit of normal in each center were higher in those with lesions (0.75 ± 0.69 versus 0.42 ± 0.29 arbitary units; p < .0001). Similar findings were observed for other biochemical markers of bone turnover, but the sensitivity of ALP and other markers in detecting lesions was poor. Asymptomatic PDB is present in about 9% of SQSTM1 mutation carriers by the fifth decade. Further follow-up of this cohort will provide important information on the natural history of early PDB and its response to treatment. © 2020 The Authors. Journal of Bone and Mineral Research published by American Society for Bone and Mineral Research.


The ribosomal RNA gene cluster in aneuploid chickens: evidence for increased gene dosage and regulation of gene expression.
In the chicken, the nucleolus organizer regions, or sites of the genes encoding 18S, 5.8S, and 28S ribosomal RNA (rRNA), map to one pair of microchromosomes that can be identified by silver nitrate cytochemistry. This nucleolar organizer chromosome also contains the major histocompatibility complex. Chickens aneuploid for this chromosome have been identified and reproduced for over seven generations. Crossing two trisomic parents results in the production of viable disomic, trisomic, and tetrasomic progeny, showing two, three, and four nucleoli and nucleolar organizers per cell, respectively. A molecular analysis of rRNA genes was undertaken to establish the gene copy numbers in the aneuploid genotypes, and to determine if elevated numbers of rRNA genes are stably maintained and inherited over multiple generations. Gene copy numbers were determined using hybridization analysis of erythrocyte DNA obtained from individuals comprising a family which segregated disomic, trisomic, and tetrasomic genotypes. The values obtained were 290, 420, and 570 rDNA repeats per cell for disomic, trisomic, and tetrasomic animals, respectively. These results provide molecular confirmation of the two aneuploid states and show that elevated gene copy numbers have been maintained over multiple generations. Fibroblasts derived from disomic and tetrasomic embryos were found to grow at similar rates in culture, and mature rRNA levels in chicken embryo fibroblasts from disomic, trisomic and tetrasomic embryos were also found to have similar levels of mature rRNA. Therefore, despite the increase in rDNA content, the level of rRNA is regulated to diploid amounts in aneuploid fibroblasts.


Phase Ib study of Buparlisib plus Trastuzumab in patients with HER2-positive advanced or metastatic breast cancer that has progressed on Trastuzumab-based therapy.
Phosphoinositide 3-kinase (PI3K)/AKT/mTOR pathway activation in patients with HER2-positive (HER2(+)) breast cancer has been implicated in de novo and acquired trastuzumab resistance. The purpose of this study was to determine the clinical activity of the PI3K inhibitor buparlisib (BKM120) in patients with HER2(+) advanced/metastatic breast cancer resistant to trastuzumab-based therapy. In the dose-escalation portion of this phase I/II study, patients with trastuzumab-resistant locally advanced or metastatic HER2(+) breast cancer were treated with daily oral doses of buparlisib and weekly intravenous trastuzumab (2 mg/kg). Dose escalation was guided by a Bayesian logistic regression model with overdose control. Of 18 enrolled patients, 17 received buparlisib. One dose-limiting toxicity of grade 3 general weakness was reported at the 100-mg/day dose level (the single-agent maximum tolerated dose) and this dose level was declared the recommended phase II dose (RP2D) of buparlisib in combination with trastuzumab. Common (>25%) adverse events included rash (39%), hyperglycemia (33%), and diarrhea (28%). The pharmacokinetic profile of buparlisib was not affected by its combination with trastuzumab. At the RP2D, there were two (17%) partial responses, 7 (58%) patients had stable disease (≥6 weeks), and the disease control rate was 75%. Pharmacodynamic studies showed inhibition of the PI3K/AKT/mTOR and RAS/MEK/ERK pathways. In this patient population, the combination of buparlisib and trastuzumab was well tolerated, and preliminary signs of clinical activity were observed. The phase II portion of this study will further explore the safety and efficacy of this combination at the RP2D. Clin Cancer Res; 20(7); 1935-45. ©2014 AACR.


A growth hormone receptor SNP promotes lung cancer by impairment of SOCS2-mediated degradation.
Both humans and mice lacking functional growth hormone (GH) receptors are known to be resistant to cancer. Further, autocrine GH has been reported to act as a cancer promoter. Here we present the first example of a variant of the GH receptor (GHR) associated with cancer promotion, in this case lung cancer. We show that the GHRP495T variant located in the receptor intracellular domain is able to prolong the GH signal in vitro using stably expressing mouse pro-B-cell and human lung cell lines. This is relevant because GH secretion is pulsatile, and extending the signal duration makes it resemble autocrine GH action. Signal duration for the activated GHR is primarily controlled by suppressor of cytokine signalling 2 (SOCS2), the substrate recognition component of the E3 protein ligase responsible for ubiquitinylation and degradation of the GHR. SOCS2 is induced by a GH pulse and we show that SOCS2 binding to the GHR is impaired by a threonine substitution at Pro 495. This results in decreased internalisation and degradation of the receptor evident in TIRF microscopy and by measurement of mature (surface) receptor expression. Mutational analysis showed that the residue at position 495 impairs SOCS2 binding only when a threonine is present, consistent with interference with the adjacent Thr494. The latter is key for SOCS2 binding, together with nearby Tyr487, which must be phosphorylated for SOCS2 binding. We also undertook nuclear magnetic resonance spectroscopy approach for structural comparison of the SOCS2 binding scaffold Ile455-Ser588, and concluded that this single substitution has altered the structure of the SOCS2 binding site. Importantly, we find that lung BEAS-2B cells expressing GHRP495T display increased expression of transcripts associated with tumour proliferation, epithelial-mesenchymal transition and metastases (TWIST1, SNAI2, EGFR, MYC and CCND1) at 2 h after a GH pulse. This is consistent with prolonged GH signalling acting to promote cancer progression in lung cancer.


Early cytokine induction by Plasmodium falciparum is not a classical endotoxin-like process.
We have investigated the widely held view that malaria parasites induce pro-inflammatory cytokines primarily through an endotoxin-like stimulatory effect on macrophages. We report that the pattern of cytokine production by non-immune human peripheral blood mononuclear cells following stimulation by Plasmodium falciparum-infected erythrocytes (Pfe) in vitro differs considerably from that induced by bacterial endotoxin. The Pfe-induced TNF response at day 1 is associated with a much higher level of IFN-gamma production and a much lower level of IL-12 p40 and IL-10 expression than a comparable endotoxin-induced TNF response. Both CD3(+) and CD14(+) populations are required for this early TNF response to Pfe, whereas the endotoxin-induced response is unaffected by depletion of the CD3(+) population. Pfe fails to stimulate the monocyte-like cell line MonoMac6 to express pro-inflammatory cytokines. These findings suggest that the early inflammatory response to malaria is critically dependent on lymphocyte subpopulations that play a lesser role in the response to bacterial endotoxin.


RNA in defense: CRISPRs protect prokaryotes against mobile genetic elements.
The CRISPR/Cas system in prokaryotes provides resistance against invading viruses and plasmids. Three distinct stages in the mechanism can be recognized. Initially, fragments of invader DNA are integrated as new spacers into the repetitive CRISPR locus. Subsequently, the CRISPR is transcribed and the transcript is cleaved by a Cas protein within the repeats, generating short RNAs (crRNAs) that contain the spacer sequence. Finally, crRNAs guide the Cas protein machinery to a complementary invader target, either DNA or RNA, resulting in inhibition of virus or plasmid proliferation. In this article, we discuss our current understanding of this fascinating adaptive and heritable defense system, and describe functional similarities and differences with RNAi in eukaryotes.


Characterization of rotavirus strains from newborns in New Delhi, India.
Between 1986 and 1993, 72% of rotavirus strains isolated from newborns at five hospitals in New Delhi, India, had long electropherotypes, subgroup II VP6 antigens, and G and P genotypes (G9P11) identical to those of prototype strain 116E. A novel strain with a G9P6 genotype, representing 13% of the isolates, was identified. These results demonstrate that G9P11 and G9P6 rotavirus strains are common in nurseries in New Delhi.


Transfer of disrupted-in-schizophrenia 1 aggregates between neuronal-like cells occurs in tunnelling nanotubes and is promoted by dopamine.
The disrupted-in-schizophrenia 1 (DISC1) gene was identified as a genetic risk factor for chronic mental illnesses (CMI) such as schizophrenia, bipolar disorder and severe recurrent depression. Insoluble aggregated DISC1 variants were found in the cingular cortex of sporadic, i.e. non-genetic, CMI patients. This suggests protein pathology as a novel, additional pathogenic mechanism, further corroborated in a recent transgenic rat model presenting DISC1 aggregates. Since the potential role of aggregation of DISC1 in sporadic CMI is unknown, we investigated whether DISC1 undergoes aggregation in cell culture and could spread between neuronal cells in a prion-like manner, as shown for amyloid proteins in neurodegenerative diseases. Co-culture experiments between donor cells forming DISC1 aggregates and acceptor cells showed that 4.5% of acceptor cells contained donor-derived DISC1 aggregates, thus indicating an efficient transfer in vitro DISC1 aggregates were found inside tunnelling nanotubes (TNTs) and transfer was enhanced by increasing TNT formation and notably by dopamine treatment, which also induces DISC1 aggregation. These data indicate that DISC1 aggregates can propagate between cells similarly to prions, thus providing some molecular basis for the role of protein pathology in CMI.


Endothelial Shc regulates arteriogenesis through dual control of arterial specification and inflammation via the notch and nuclear factor-κ-light-chain-enhancer of activated B-cell pathways.
Arteriogenesis, the shear stress-driven remodeling of collateral arteries, is critical in restoring blood flow to ischemic tissue after a vascular occlusion. Our previous work has shown that the adaptor protein Shc mediates endothelial responses to shear stress in vitro. To examine the role of the adaptor protein Shc in arteriogenesis and endothelial-dependent responses to shear stress in vivo. Conditional knockout mice in which Shc is deleted from endothelial cells were subjected to femoral artery ligation. Hindlimb perfusion recovery was attenuated in Shc conditional knockout mice compared with littermate controls. Reduced perfusion was associated with blunted collateral remodeling and reduced capillary density. Bone marrow transplantation experiments revealed that endothelial Shc is required for perfusion recovery because loss of Shc in bone marrow-derived hematopoietic cells had no effect on recovery. Mechanistically, Shc deficiency resulted in impaired activation of the nuclear factor κ-light-chain-enhancer of activated B-cell-dependent inflammatory pathway and reduced CD45⁺ cell infiltration. Unexpectedly, Shc was required for arterial specification of the remodeling arteriole by mediating upregulation of the arterial endothelial cell marker ephrinB2 and activation of the Notch pathway. In vitro experiments confirmed that Shc was required for shear stress-induced activation of the Notch pathway and downstream arterial specification through a mechanism that involves upregulation of Notch ligands delta-like 1 and delta-like 4. Shc mediates activation of 2 key signaling pathways that are critical for inflammation and arterial specification; collectively, these pathways contribute to arteriogenesis and the recovery of blood perfusion.


Predicting the outcome of psychotherapy. findings of the Penn Psychotherapy Project.
Our study of predictability of outcomes of psychotherapy used predictions of two kinds: (1) direct predictions by patients, therapists, and clinical observers; and (2) predictive measures derived from the same sources. Seventy-three nonpsychotic patients were treated in psychoanalytically oriented psychotherapy (mean, 44 sessions). Two thirds of the therapists were residents in psychiatry; one third were more experienced. The two main composite outcome measures, measured at termination, were Raw Gain (residualized) and Rated Benefits, which intercorrelated at .76. Most patients improved and showed a considerable range of benefits. The clinical observers' direct predictions of Rated Benefits were highest (.27, P less than 905). The success of the predictive measures were generally insignificant, and the best of them were in the .2 to .3 range meaning that only 5% to 10% of the outcome variance was predicted. The Prognostic Index Interview variables did the best (eg, emotional freedom composite, .30; a crossvalidation for 30 patients was .39 (P less than .05). Neither the therapist measures nor the early psychotherapy session measures predicted significantly. Reanalysis of the similar Chicago Counseling Center study, in our terms, showed a similar low level of prediction success, eg, adequacy of functioning, marital status match, and length of treatment predicted significantly in both studies.


The use of modeling to understand the impact of screening on U.S. mortality: examples from mammography and PSA testing.
Surveillance data represent a vital resource for understanding the impact of cancer control interventions on the population cancer burden. However, population cancer trends are a complex product of many factors, and estimating the contribution of any one of these factors can be challenging. Surveillance modeling is a technique for estimating the contribution of one or more interventions of interest to trends in disease incidence and mortality. In this article, we present several approaches to surveillance modeling of cancer screening interventions. We classify models as biological or epidemiological, depending on whether they model the full unobservable aspects of disease onset and progression, or models which reduce the complex process to simpler terms by summarizing portions of the disease process using mostly observed population level measures. We also describe differences between macrolevel models, microsimulation models and mechanistic models. We discuss procedures for model calibration and validation, and methods for presenting model results which are robust with respect to certain types of biased model estimates. As examples, we present several models of the impact of mammography screening on breast cancer mortality, and PSA screening on prostate cancer mortality. Both these examples are appropriate uses of surveillance modeling, even though for mammography there is extensive (although somewhat controversial) randomized trial evidence, whereas for PSA this biomarker has seen extensive use as a screening test prior to any controlled trial evidence of its efficacy. Some of the models presented here were developed as part of the National Cancer Institute's Cancer Intervention and Surveillance Modeling Network.