Deducing the functional characteristics of the human selenoprotein SELK from the structural properties of its intrinsically disordered C-terminal domain.
The intrinsically disordered proteins (IDPs) cannot be described by a single structural representation but, due to their high structural fluctuation, through conformational ensembles. Certainly, molecular dynamics (MD) simulations represent a useful tool to study their different conformations capturing the conformational distribution. Our group is focusing on the structural characterization of proteins belonging to the seleno-proteome due to their involvement in cancer. They present disordered domains central for their biological function, and, in particular, SELK is a single-pass transmembrane protein that resides in the endoplasmic reticulum membrane (ER) with a C-terminal domain exposed to the cytoplasm that is known to interact with different components of the endoplasmic reticulum associated to the protein degradation (ERAD) pathway. This protein is found to be up-expressed in hepatocellular carcinoma and in other cancers. In this work we performed a detailed analysis of the C-terminal domain sequence of SELK and discovered that it is characterized by many prolines, and four negatively and eleven positively charged residues, which are crucial for its biological activity. This region can be considered as a weak polyelectrolyte and, specifically, a polycation, with high disordered propensity and different phosphorylation sites dislocated along the sequence. Then, we modeled its three-dimensional structure by performing MD simulations in water at neutral pH to analyze the structural stability as well as to identify the presence of HUB residues that play a key structural role as evidenced by the residue-residue interaction network analysis. Through this approach, we demonstrate that the C-terminal domain of SELK (i) presents a poor content of regular secondary structure elements, (ii) is dynamically stabilized by a network of intra-molecular H-bonds and H-bonds with water molecules, (iii) is highly fluctuating and, therefore, can be described only through a conformational ensemble, where we evidenced a distribution of equilibrium conformers which continuously inter-change their conformations. Finally to verify the specific role played by the negative charges, we also performed MD simulations at acidic pH. Overall, all the obtained results evidenced that SELK has the dynamic structural features to be defined as a HUB protein able to interact with multiple members. Therefore, considering the possible role that this protein can have in cancer development and progression, it can represent a target for drug design studies.


Xeroderma pigmentosum: low prevalence of germline XPA mutations in a Brazilian XP population.
Xeroderma pigmentosum (XP) is a rare autosomal recessive disorder characterized by DNA repair defects that cause photophobia, sunlight-induced cancers, and neurodegeneration. Prevalence of germline mutations in the nucleotide excision repair gene XPA vary significantly in different populations. No Brazilian patients have been reported to carry a germline mutation in this gene. In this study, the germline mutational status of XPA was determined in Brazilian patients exhibiting major clinical features of XP syndrome. The study was conducted on 27 unrelated patients from select Brazilian families. A biallelic inactivating transition mutation c.619C>T (p.Arg207Ter) was identified in only one patient with a history of neurological impairment and mild skin abnormalities. These findings suggest that XP syndrome is rarely associated with inherited disease-causing XPA mutations in the Brazilian population. Additionally, this report demonstrates the effectiveness of genotype-phenotype correlation as a valuable tool to guide direct genetic screening.


Inactivation of human immunodeficiency virus type 1 by nonoxynol-9, C31G, or an alkyl sulfate, sodium dodecyl sulfate.
A highly desirable approach to prevention of human immunodeficiency virus type 1 (HIV-1) transmission during sexual intercourse is the development of nontoxic, topical, broad spectrum microbicides effective against transmission of cell-associated and cell-free virus. Toward this end, the HIV-1 inactivation potential of surface active agents C31G and an alkyl sulfate, sodium dodecyl sulfate (SDS) was assessed. Because of its extensive use as a microbicidal agent, nonoxynol-9 (N-9) was used as a reference against which C31G and SDS were compared. Viral inactivation was measured using HIV-1 LTR-beta-galactosidase indicator cells (expressing CD4 or CD4/CCR5) derived from HeLa cells, a cell line of human cervical adenocarcinoma origin. In experiments which examined inactivation of cell-free HIV-1, C31G was generally more effective than N-9. Viral inactivation by SDS occurred at twice the concentration necessary to achieve similar levels of inactivation using either N-9 or C31G. Using HeLa and HeLa-derived cells in cytotoxicity studies, it was demonstrated that SDS is as much as 11 and five times less cytotoxic than N-9 or C31G, respectively, during 48 h of continuous exposure. SDS (unlike C31G and N-9) can inactivate non-enveloped viruses such as human papillomavirus (HPV) [Howett, M.K., Neely, E.B., Christensen, N.D., Wigdahl, B., Krebs, F.C., Malamud, D., Patrick, S.D., Pickel, M.D., Welsh, P.A., Reed, C.A., Ward, M.G., Budgeon, L.R., Kreider, J.W., 1999. A broad-spectrum microbicide with virucidal activity against sexually transmitted viruses. Antimicrob. Agents Chemother. 43(2), 314-321]. Since addition of SDS to C31G or N-9 may make the resulting microbicidal mixtures broadly effective against both enveloped and non-enveloped viruses, several surface active agent combinations were evaluated for their abilities to inactivate HIV-1. Addition of SDS to either C31G or N-9 resulted in mixtures that were only slightly less effective than equivalent concentrations of C31G or N-9 alone. To investigate inactivation of cell-associated infectivity, HIV-1 IIIB-infected SupT1 cells were treated with N-9, C31G, or SDS. Inactivation of cell-associated infectivity required higher microbicide concentrations than were needed for inactivation of cell-free virus. However, the relative activities of N-9, C31G, or SDS were similar to those seen in assays of inactivation using cell-free virus. These studies suggest that C31G and SDS may be attractive candidates for human trials as topical microbicides effective against HIV-1 transmission since both function at concentrations that provide effective viral inactivation with low levels of cytotoxicity. SDS microbicides (used alone or with other microbicides) may provide the added advantage of protection from HPV infection.


Arterial reconstruction of the brachiocephalic trunk and the subclavian arteries. 10 years' experience with a follow-up study.
During a 10-year period, 60 patients were operated on for occlusive lesions in the brachiocephalic trunk and/or the subclavian arteries proximal to the vertebral artery. Angiography showed 68 occlusive lesions, of which 64 were treated surgically. Of these, 47 exhibited subclavian steal. Sixty-three reconstructive procedures were performed together with two arterial ligations (four patients were operated on bilaterally and one patient was operated on twice on the same side). There were 30 transthoracic procedures, essentially endarterectomies, and 35 supraclavicular procedures, mostly carotid-subclavian bypasses and transpositions of the subclavian artery to the common carotid artery. One patient died (1.5 per cent). Serious complications occurred in additional four cases. In the follow-up study data on all the patients were available. During the follow-up period, ranging from four to 124 months (mean 43 months) seven patients died - none of cerebral ischemia. Based on blood pressure recordings and in doubtful cases, angiography, we found a cumulated five-year patency rate of 76 per cent. Seventy per cent of the patients considered their condition after operation to be improved, 26 per cent unchanged and three per cent worsened. Twice as many patients could work after the operation as before. The cumulative survival rate was found to be somewhat lower than that of a normal population.


Critical rearing parameters of Tetrastichus planipennisi (Hymenoptera: Eulophidae) as affected by host plant substrate and host-parasitoid group structure.
In laboratory assays, we evaluated the potential impact of host plant substrate types, host-parasitoid group sizes (densities), and parasitoid-to-host ratios on select fitness parameters of the larval endoparasitoid Tetrastichus planipennisi Yang (Hymenoptera: Eulophidae), newly introduced for biological control of emerald ash borer, Agrilus planipennis Fairmaire (Coleoptera: Buprestidae), in the United States. Results from our study showed that offspring production and critical fitness parameters (body size and sex ratio) of T. planipennisi from parasitized emerald ash borer larvae are significantly influenced by host plant substrate type, host-parasitoid group size, parasitoid-to-host ratio, or a combination in the primary exposure assay. The number of both female and male T. planipennisi progeny was significantly greater when emerald ash borer larvae were inserted into tropical ash [Fraxinus uhdei (Wenz.) Lingelsh.] logs rather than green ash (Fraxinus pensylvanica Marshall). When maintained at a constant 1:1 parasitoid-to-host ratio, assays with larger host-parasitoid group sizes (3:3-12:12) produced significantly greater numbers of both male and female offspring per parental wasp compared with those with the single host-parasitoid (1:1) group treatment. As the parasitoid-to-host ratio increased from 1:1 to 8:1 in the assay, the average brood size (number of offspring per parasitized emerald ash borer larva) increased significantly, whereas the average brood sex ratio (female to male) changed from being female-biased (6:1) to male-biased (1:2); body size of female offspring as measured by the length of ovipositor and left hind tibia also was reduced significantly. Based on these findings, we suggest that the current method of rearing T. planipennisi with artificially infested-emerald ash borer larvae use the tropical ash logs for emerald ash borer insertion, a larger (> or = 3:3) host-parasitoid group size and 1:1 parasitoid-to-host ratio in the primary parasitoid exposure assays.


[Lifetime diagnosis of prion transmissive encephalopathies].
An original method for life-time diagnosis of Greutzfeldt-Jakob's disease, a neurodegenerative disease belonging to transmissive spongiform encephalopathies, is proposed. It consists in inoculation of the examined biological materials (blood serum and clot) into a continuous culture of the rat Gasser's node neurinoma, followed by passages and study of the inoculated culture by morphological methods and electron microscopy.


IMPOSE (IMProving Outcomes after Sepsis)-the effect of a multidisciplinary follow-up service on health-related quality of life in patients postsepsis syndromes-a double-blinded randomised controlled trial: protocol.
Patients post sepsis syndromes have a poor quality of life and a high rate of recurring illness or mortality. Follow-up clinics have been instituted for patients postgeneral intensive care but evidence is sparse, and there has been no clinic specifically for survivors of sepsis. The aim of this trial is to investigate if targeted screening and appropriate intervention to these patients can result in an improved quality of life (Short Form 36 health survey (SF36V.2)), decreased mortality in the first 12 months, decreased readmission to hospital and/or decreased use of health resources. 204 patients postsepsis syndromes will be randomised to one of the two groups. The intervention group will attend an outpatient clinic two monthly for 6 months and receive screening and targeted intervention. The usual care group will remain under the care of their physician. To analyse the results, a baseline comparison will be carried out between each group. Generalised estimating equations will compare the SF36 domain scores between groups and across time points. Mortality will be compared between groups using a Cox proportional hazards (time until death) analysis. Time to first readmission will be compared between groups by a survival analysis. Healthcare costs will be compared between groups using a generalised linear model. Economic (health resource) evaluation will be a within-trial incremental cost utility analysis with a societal perspective. Ethical approval has been granted by the Royal Brisbane and Women's Hospital Human Research Ethics Committee (HREC; HREC/13/QRBW/17), The University of Queensland HREC (2013000543), Griffith University (RHS/08/14/HREC) and the Australian Government Department of Health (26/2013). The results of this study will be submitted to peer-reviewed intensive care journals and presented at national and international intensive care and/or rehabilitation conferences. Australian and New Zealand Clinical Trials Registry ACTRN12613000528752.


The analysis of the relationships of Korean outbound tourism demand: Jeju Island and three international destinations.
This paper investigates the determinants of the relationship among Korean outbound tourism demand for Jeju Island and three other Asian island countries using the multivariate generalized autoregressive conditional heteroskedasticity (MGARCH) and Vector Error Correction (VEC) models. It is found that pairwise conditional correlations among tourism demand for Jeju, Thailand, Singapore and the Philippines are not constant but time-varying. Estimated conditional correlations among Jeju and the three Asian countries are negative over some time periods. This implies that the three Asian countries are substitutes for Jeju in certain specific time horizons. The VEC model is used to investigate the short-run and long-run dynamic relationships and the results reveal that Industrial Production Index and real exchange rates had the positive or negative impact on conditional correlations of tourism demand for these destinations. Tourism policy implications are discussed for managing tourism demand for these destinations.


Fibrinogen binds to integrin alpha(5)beta(1) via the carboxyl-terminal RGD site of the Aalpha-chain.
Fibrinogen interactions with vascular endothelial cells are implicated in various physiological and pathophysiological events, including angiogenesis and wound healing. We have shown previously that integrin alpha(5)beta(1) is a fibrinogen receptor on endothelial cells [Suehiro, K., Gailit, J., and Plow, E.F. (1997) J. Biol. Chem. 272, 5360-5366]. In the present study, we have characterized fibrinogen interactions with purified alpha(5)beta(1) and have identified the recognition sequence in fibrinogen for alpha(5)beta(1). The binding of fibrinogen to immobilized alpha(5)beta(1) was selectively supported by Mn(2+). Fibrinogen bound to purified alpha(5)beta(1) in a time-dependent, specific, and saturable manner in the presence of Mn(2+), and the binding was blocked completely by Arg-Gly-Asp (RGD)-containing peptides and by anti-alpha(5) and anti-alpha(5)beta(1) monoclonal antibodies. A monoclonal antibody directed to the C-terminal RGD sequence at Aalpha572-574 significantly inhibited the binding of fibrinogen to alpha(5)beta(1), whereas monoclonal antibodies directed to either the N-terminal RGD sequence at Aalpha95-97 or the C-terminus of the gamma-chain did not. Furthermore, substituting RGE for RGD at position Aalpha95-97 in recombinant fibrinogen had a minimal effect on binding, whereas substituting RGE for RGD at position Aalpha572-574 decreased binding by 90%. These results demonstrate that the C-terminal RGD sequence at Aalpha572-574 is required for the interaction of fibrinogen with alpha(5)beta(1).


Vancomycin: we can't get there from here.
We sought to characterize the pharmacodynamic profile of the more intensive vancomycin dosing regimens currently used in response to the recent vancomycin guidelines. A series of Monte Carlo simulations was performed for vancomycin regimens ranging from .5 g intravenous (IV) Q12H to 2 g IV Q12H. The probability of achieving an AUC/MIC ratio ≥ 400 for each dosing regimen was calculated for minimum inhibitory concentrations (MICs) from .5 to 2 mg/L. The risk of nephrotoxicity for each regimen was derived from a previously published vancomycin trough-nephrotoxicity logistic regression function. Restricted analyses were performed that only included subjects with troughs between 15 and 20 mg/L. At a MIC of 2 mg/L, even the most aggressive dosing regimen considered (2 g every 12 h) only yielded a probability of target attainment (PTA) of 57% while generating a nephrotoxicity probability upward of 35%(.) At a MIC of 1 mg/L, ≥3 g per day provided PTA in excess of 80% but were associated with unacceptable risks of nephrotoxicity. In the restricted analyses of subjects with troughs between 15 and 20 mg/L, all regimens produced a PTA of 100% at MICs ≤1 mg/L. The PTA was variable among the regimens at a MIC of 2 mg/L and was highly dependent on the total daily dose administered. This study indicates that vancomycin may not be useful for treating serious methicillin-resistant Staphylococcus aureus (MRSA) infections with MIC values > 1 mg/L where PTA is questionable. Since an AUC/MIC ratio ≥ 400 is target associated with efficacy, one should consider incorporating computation of AUC when monitoring vancomycin.


Function of the mitochondrial outer membrane as a diffusion barrier in health and diseases.
The mitochondrial outer membrane separates the intermembrane space from the cytosol. The whole exchange of metabolites, cations and information between mitochondria and the cell occurs through the outer membrane. Experimental evidence is reviewed supporting the hypothesis of dynamic ADP compartmentation within the intermembrane space. The outer membrane creates a diffusion barrier for small molecules (adenine nucleotides, creatine phosphate, creatine etc.) causing rate-dependent concentration gradients as a prerequisite for the action of ADP shuttles via creatine kinases or adenylate kinases. If the outer membrane becomes leaky, cytochrome c and apoptosis-inducing factor can be released, leading to apoptosis, and as a bioenergetic consequence the cytosolic phosphorylation potential decreases. Leaky outer membranes can be detected in saponin-skinned fibres with spectrophotometric and oxygraphic methods. This is of special interest in respect to acute impairment of mitochondria during ischaemia/reperfusion.


Decreased VEGF expression and microvascular density, but increased HIF-1 and 2α accumulation and EPO expression in chronic moderate hyperoxia in the mouse brain.
Normal brain function is dependent on continuous and controlled oxygen delivery. Chronic moderate hypoxia leads to angiogenesis, suggesting a modulatory role for oxygen in determining capillary density. The objective of this study was to determine physiologic and brain angiogenic adaptational changes during chronic moderate normobaric hyperoxia in mice. Four-month old C56BL/6J mice were kept in a normobaric chamber at 50% O(2) for up to 3 weeks. Normoxic littermates were kept in the same room outside the chamber. Freshly collected or fixed brain specimens were analyzed by RT-PCR, Western blot analysis and immunohistochemistry. Results show accumulation of hypoxia inducible factors 1 and 2α (HIF-1 and 2α), and increased expression of erythropoietin (EPO), cyclooxygenase-2 (COX-2) and angiopoietin-2 (Ang-2). Conversely, vascular endothelial growth factor (VEGF), and VEGF receptor-2 (KDR/Flk-1), Peroxisome proliferator-activated receptor gamma coactivator 1-α (PGC-1α) and prolylhydroxylase-2 (PHD-2) expressions were decreased. VEGF mRNA level was diminished but there was no change in HIF-1α mRNA and von Hippel Lindau E3 ubiquitin ligase (VHL) protein expression. Microvascular density was significantly diminished by the end of the 3rd week of hyperoxia. Overall, our results are: (1) increased expression of the potent neuroprotective molecule, EPO; (2) diminished expression of the potent angiogenic factor, VEGF; and (3) decreased microvascular density. We can, therefore, conclude that brain microvascular density can be controlled by HIF-independent mechanisms, and that brain capillary density is a continuously adjusted variable with tissue oxygen availability as one of the controlling modulators.


Recombinant hirudin for prevention of experimental postoperative intraocular fibrin.
To determine the efficacy of a specific antithrombin agent (recombinant desulphatohirudin variant 1 [Revasc, Ciba-Geigy, Ltd., Basel, Switzerland]) administered in the infusion fluid to prevent early postoperative fibrin formation in a rabbit lensectomy and vitrectomy model. Standard fragmatome lensectomies and core vitrectomies were performed prospectively in a masked fashion on ten control eyes with lactated Ringer's infusion and on ten eyes treated with 10 microgram of recombinant hirudin/ml in the infusate. The amounts of fibrin and hemorrhage were graded in a masked fashion by using slit-lamp examination and indirect ophthalmoscopy on postoperative days 1 through 5 and on day 7. The difference in the mean grade of fibrin formed on the first postoperative day in the eyes treated with recombinant hirudin (mean, 0.9) in relation to the mean grade of fibrin in the control eyes (mean, 3.5) was statistically significant (P = .004). This difference was also significant on the second postoperative day (P = .01). None of the treated eyes developed intraoperative or postoperative hemorrhage. Recombinant desulphatohirudin variant 1 is an effective inhibitor of postoperative fibrin formation in a rabbit model and is not associated with an increased risk of intraoperative or postoperative bleeding at the tested dose. This drug may be a useful adjunct in vitreous surgery for both proliferative vitreoretinopathy and the complications of proliferative diabetic retinopathy.


Influence of automatic exposure on the estimation of seasonal variations in leaf area index measured by digital hemispherical photograph.
Automatic exposure is one of the important error sources during measurement of leaf area index (LAI) by digital hemispherical photography (DHP). This study was conducted in a mixed broadleaved-Korean pine (Pinus koraiensis) forest, a secondary birch (Betula platyphylla) forest, a Korean pine plantation and a Dahurian larch (Larix gmelinii) plantation in the Xiaoxing'an Mountains. LAI was measured using DHP and LAI-2200 plant canopy analyzer in the middle of June to September. We compared LAI values measured through these two methods, and then tested whether the forest type and study period had a significant influence on the correlations between the measured values of those two methods. We constructed empirical models for correcting the errors caused by automatic exposure for LAI values measured through DHP at different study periods in different forest types. The results showed that LAI from DHP was underestimated by 20%-49% rela-tive to that from LAI-2200 in four study periods of the four forest types. Forest type had no significant effect on the construction of empirical models between these two measuring methods of LAI, whereas study period showed significant effects. Two classified empirical models (A and B) were constructed, which were suitable for correcting the LAI from DHP in June and September, July and August in four forest types, respectively. After being corrected by the classified empirical models, LAI from DHP of the four forest types increased by 45%-79%, and the measurement accuracy could be improved to 83%-94%. Classified empirical models between LAI from DHP and LAI-2200 could effectively correct the influence of automatic exposure on DHP and greatly improve its mea-surement accuracy, and provide a technical support for rapid and effective measurement of seasonal changes of LAI in different forest types.


Predictive Value of Matrix Metalloproteinases and Their Inhibitors for Mortality in Septic Patients: A Cohort Study.
Over 170 biomarkers are being investigated regarding their prognostic and diagnostic accuracy in sepsis in order to find new tools to reduce morbidity and mortality. Matrix metalloproteinases (MMPs) and their inhibitors have been recently studied as promising new prognostic biomarkers in patients with sepsis. This study is aimed at determining the utility of several cutoff points of these biomarkers to predict mortality in patients with sepsis. A multicenter, prospective, analytic cohort study was performed in the metropolitan area of Bucaramanga, Colombia. A total of 289 patients with sepsis and septic shock were included. MMP-9, MMP-2, tissue inhibitor of metalloproteinase 1 (TIMP-1), TIMP-2, TIMP-1/MMP-9 ratio, and TIMP-2/MMP-2 ratio were determined in blood samples. Value ranges were correlated with mortality to estimate sensitivity, specificity, positive predictive value, negative predictive value, and area under the receiving operating characteristic curve. Sensitivity ranged from 33.3% (MMP-9/TIMP-1 ratio) to 60.6% (TIMP-1) and specificity varied from 38.8% (MMP-2/TIMP-2 ratio) to 58.5% (TIMP-1). As for predictive values, positive predictive value range was from 17.5% (MMP-9/TIMP-1 ratio) to 70.4% (MMP-2/TIMP-2 ratio), whereas negative predictive values were between 23.2% (MMP-2/TIMP-2 ratio) and 80.9% (TIMP-1). Finally, area under the curve scores ranged from 0.31 (MMP-9/TIMP-1 ratio) to 0.623 (TIMP-1). Although TIMP-1 showed higher sensitivity, specificity, and negative predictive value, with a representative population sample, we conclude that none of the evaluated biomarkers had significant predictive value for mortality.


Micro-incision vitrectomy surgery for primary rhegmatogenous retinal detachments with posterior vitreous detachments in elderly patients: Preoperative characteristics and surgical outcomes.
To assess the preoperative characteristics and surgical outcomes of using micro-incision vitrectomy surgery (MIVS) to treat RRD with posterior vitreous detachment (PVD) in an older and a younger patient group. This retrospective cohort study included 407 eyes from 397 patients with primary RRD with PVD who were consecutively treated in our hospital from February 2016 to February 2020. PVD was diagnosed clinically by the presence of a Weiss ring, or was diagnosed morphologically via optical coherence tomography and subsequently confirmed during surgery. The main outcome measures were preoperative RRD characteristics, best-corrected visual acuity (BCVA), and postoperative complications. Data were analysed from 55 eyes in the elderly group (age 70 and older), and 352 eyes in the young group (age 69 and younger). There was no significant inter-group difference in the initial reattachment rate. Preoperative characteristics indicated that elderly patients had a significantly lower rate of phakic eyes, shorter mean axial length, lower lattice incidence, and longer time spans from onset to surgery. There were no significant between-group differences in the incidence of the following complications: fibrin formation, intraocular pressure elevation, epi-retinal membrane on the macula, intraocular lens optic capture, proliferative vitreoretinopathy, and vitreous haemorrhage. While the elderly patients had significant postoperative improvements in BCVA, these improvements were significantly lower than those of the younger patients. This study highlighted the characteristics and surgical outcomes of MIVS in elderly patients with RRD. Although the time from onset to surgery was longer, MIVS still can be performed safely to improve older patients' postoperative BCVA.


Epigenome-wide association studies of prenatal maternal mental health and infant epigenetic profiles: a systematic review.
Prenatal stress and poor maternal mental health are associated with adverse offspring outcomes; however, the biological mechanisms are unknown. Epigenetic modification has linked maternal health with offspring development. Epigenome-wide association studies (EWAS) have examined offspring DNA methylation profiles for association with prenatal maternal mental health to elucidate mechanisms of these complex relationships. The objective of this study is to provide a comprehensive, systematic review of EWASs of infant epigenetic profiles and prenatal maternal anxiety, depression, or depression treatment. We conducted a systematic literature search following PRISMA guidelines for EWAS studies between prenatal maternal mental health and infant epigenetics through May 22, 2023. Of 645 identified articles, 20 fulfilled inclusion criteria. We assessed replication of CpG sites among studies, conducted gene enrichment analysis, and evaluated the articles for quality and risk of bias. We found one repeated CpG site among the maternal depression studies; however, nine pairs of overlapping differentially methylatd regions were reported in at least two maternal depression studies. Gene enrichment analysis found significant pathways for maternal depression but not for any other maternal mental health category. We found evidence that these EWAS present a medium to high risk of bias. Exposure to prenatal maternal depression and anxiety or treatment for such was not consistently associated with epigenetic changes in infants in this systematic review and meta-analysis. Small sample size, potential bias due to exposure misclassification and statistical challenges are critical to address in future efforts to explore epigenetic modification as a potential mechanism by which prenatal exposure to maternal mental health disorders leads to adverse infant outcomes.


A cost-consequences analysis of a midwifery group practice for Aboriginal mothers and infants in the top end of the Northern Territory, Australia.
to compare the cost-effectiveness of two models of service delivery: Midwifery Group Practice (MGP) and baseline cohort. a retrospective and prospective cohort study. a regional hospital in Northern Territory (NT), Australia. baseline cohort included all Aboriginal mothers (n=412), and their infants (n=416), from two remote communities who gave birth between 2004 and 2006. The MGP cohort included all Aboriginal mothers (n=310), and their infants (n=315), from seven communities who gave birth between 2009 and 2011. The baseline cohort mothers and infant's medical records were retrospectively audited and the MGP cohort data were prospectively collected. All the direct costs, from the Department of Health (DH) perspective, occurred from the first antenatal presentation to six weeks post partum for mothers and up to 28 days post births for infants were included for analysis. analysis was performed with SPSS 19.0 and Stata 12.1. Independent sample of t-tests and χ2 were conducted. women receiving MGP care had significantly more antenatal care, more ultrasounds, were more likely to be admitted to hospital antenatally, and had more postnatal care in town. The MGP cohort had significantly reduced average length of stay for infants admitted to Special Care Nursery (SCN). There was no significant difference between the two cohorts for major birth outcomes such as mode of birth, preterm birth rate and low birth weight. Costs savings (mean A$703) were found, although these were not statistically significant, for women and their infants receiving MGP care compared to the baseline cohort. for remote dwelling Aboriginal women of all risk who travelled to town for birth, MGP was likely to be cost effective, and women received better care and resulting in equivalent birth outcomes compared with the baseline maternity care.


Protocol for a national prevalence study of advance care planning documentation and self-reported uptake in Australia.
Advance care planning (ACP) is a process between a person, their family/carer(s) and healthcare providers that supports adults at any age or stage of health in understanding and sharing their personal values, life goals and preferences regarding future medical care. The Australian government funds a number of national initiatives aimed at increasing ACP uptake; however, there is currently no standardised Australian data on formal ACP documentation or self-reported uptake. This makes it difficult to evaluate the impact of ACP initiatives. This study aims to determine the Australian national prevalence of ACP and completion of Advance Care Directives (ACDs) in hospitals, aged care facilities and general practices. It will also explore people's self-reported use of ACP and views about the process. Researchers will conduct a national multicentre cross-sectional prevalence study, consisting of a record audit and surveys of people aged 65 years or more in three sectors. From 49 participating Australian organisations, 50 records will be audited (total of 2450 records). People whose records were audited, who speak English and have a decision-making capacity will also be invited to complete a survey. The primary outcome measure will be the number of people who have formal or informal ACP documentation that can be located in records within 15 min. Other outcomes will include demographics, measure of illness and functional capacity, details of ACP documentation (including type of document), location of documentation in the person's records and whether current clinical care plans are consistent with ACP documentation. People will be surveyed, to measure self-reported interest, uptake and use of ACP/ACDs, and self-reported quality of life. This protocol has been approved by the Austin Health Human Research Ethics Committee (reference HREC/17/Austin/83). Results will be submitted to international peer-reviewed journals and presented at international conferences. ACTRN12617000743369.


Dietary supplements, guideline alignment and biochemical nutrient status in pregnancy: Findings from the Queensland Family Cohort pilot study.
In high-income nations, multiple micronutrient (MMN) supplementation during pregnancy is a common practice. We aimed to describe maternal characteristics associated with supplement use and daily dose of supplemental nutrients consumed in pregnancy, and whether guideline alignment and nutrient status are related to supplement use. The Queensland Family Cohort is a prospective, Australian observational longitudinal study. Maternal characteristics, nutrient intake from food and supplements, and biochemical nutrient status were assessed in the second trimester (n = 127). Supplement use was reported by 89% of participants, of whom 91% reported taking an MMN supplement. Participants who received private obstetric care, had private health insurance and had greater alignment to meat/vegetarian alternatives recommendations were more likely to report MMN supplement use. Private obstetric care and general practitioner shared care were associated with higher daily dose of supplemental nutrients consumed compared with midwifery group practice. There was high reliance on supplements to meet nutrient reference values for folate, iodine and iron, but only plasma folate concentrations were higher in MMN supplement versus nonsupplement users. Exceeding the upper level of intake for folic acid and iron was more likely among combined MMN and individual supplement/s users, and associated with higher plasma concentrations of the respective nutrients. Given the low alignment with food group recommendations and potential risks associated with high MMN supplement use, whole food diets should be emphasized. This study confirms the need to define effective strategies for optimizing nutrient intake in pregnancy, especially among those most vulnerable where MMN supplement use may be appropriate.


Histamine H2-receptor antagonist ranitidine protects against neural death induced by oxygen-glucose deprivation.
Administration of histamine receptor antagonists has been reported to produce contradictory results, either reducing or increasing neural damage induced by ischemia. In this study, we investigated the neuroprotective effects of histamine H2-receptor antagonists in an "in vitro" model of ischemia. Cultured rat brain cortical neurons were exposed to oxygen-glucose deprivation (OGD) in the presence or absence of different histaminergic drugs. Cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide reduction assay. Necrosis and apoptosis were quantified by staining cells with propidium iodide and Hoechst 33258. Caspase 3 activation was determined by immunocytochemistry and Western blot. Pretreatment with H2 antagonists effectively reduced neuronal cell death induced by OGD. Ranitidine decreased the number of necrotic and apoptotic cells. Caspase 3 activation and alteration of the neuronal cytoskeleton were also prevented by ranitidine pretreatment. The neuroprotective effect of ranitidine was still evident when added 6 hours after OGD. H2-receptor antagonists protected against OGD-induced neuronal death. Ranitidine attenuated cell death even when administered after OGD. These data suggest that this drug, which is currently used for the treatment of gastric ulcers, may be useful in promoting recovery after ischemia.


Intracellular localization, isolation and characterization of two distinct varieties of superoxide dismutase from Neurospora crassa.
1. Neurospora crassa was found to contain two distinct superoxide dismutases. 2. Most of the activity is associated with the cytosolic fraction and was shown to be the Cu/Zn-containing form of the protein. 3. Mitochondria isolated from Neurospora crassa showed two distinct superoxide dismutases: a cyanide-sensitive Cu/Zn-containing protein and a cyanide-insensitive form which probably contains manganese. 4. Localization experiments, using selective marker enzymes and digitonin fractionation, indicated that the cyanide-sensitive form is localized in the intermembrane space, whereas the cyanide-insensitive form is confined to the mitochondrial matrix space. 5. The cytosolic Cu/Zn-containing superoxide dismutase was isolated in high yields and extensively characterized by using e.p.r. spectroscopy, isoelectric focusing and analytical ultracentrifugation. 6. E.p.r. spectroscopy was used to monitor changes in the copper environment of the native protein after the addition of a number of potential inhibitors and after high-pH treatment. 7. Both of the cyanide-sensitive Cu/Zn-containing enzymes (cytosolic and mitochondrial) appeared to have identical properties which in turn were different from the cyanide-insensitive enzyme. 8. It is probable that the cyanide-insensitive enzyme was not previously detected, owing to its low amount (less than 10% of the total activity), greater lability than the cyanide-sensitive enzyme and the necessity of obtaining a mitochondrial-enriched fraction before its isolation.


Coins in microfluidics: From mere scale objects to font of inspiration for microchannel circuits.
The fabrication of microfluidic chips remains a complex and expensive process requiring specific equipment and protocols, often if not always limited to the most privileged laboratories. As an alternative to the most sophisticated methods, the present paper describes the fabrication of microfluidic chips by an approach that uses coins as positive master for the rapid production of multigeometry chips. All steps of chip production were carried out using inexpensive approaches by low-cost chemicals and equipment. The chips were validated by different "classic" microfluidic tasks, such as hydrodynamic focusing, droplets generation, micromixing, and on-chip cell culture. The use of coins is not only an efficient method for rapid prototyping but also represents an inspiring possibility for the design of new microfluidic chips. Finally, coin-inspired chips could represent a laboratory experiment doable at a high school level.


Neutralization of foot-and-mouth disease virus. II. Further parameters related to the sensitization of the 140S virion by antibody.
The reaction of foot-and-mouth disease virus (FMDV) with 12S subunit/140S virion cross-reactive (sensitizing) antibody was studied in order to elucidate the requirements for neutralization versus sensitization. The presence of sensitizing antibody in immune serum caused an atypical in vitro neutralization response curve and a non-neutralized fraction. Cell-associated (cytophilic) antibody was not present in the system. Dissociation of the immune complex was not a factor and sensitized virus adsorbed to host cells via the regular virus receptor site(s). This finding led to the conclusion that sensitizing antibody is specific for non-critical sites. Dosing of the neutralization reaction mixtures with fractionated antibody of alternative antigenic specificities had an antagonistic effect on the neutralization response, suggesting steric hindrance. Cell receptor sites were a factor in sensitization since different host systems had different susceptibilities for sensitized antigen. The results suggest that in vitro neutralization of FMDV requires the attachment of multiple antibody molecules as proposed by the multi-hit theory of neutralization. The in vitro measurement of serum neutralizing activity as an indication of the in vivo immune response is discussed.


Antioxidant/stress response in mouse epidermis following exposure to nitrogen mustard.
Nitrogen mustard (NM) is a highly reactive bifunctional alkylating agent that induces inflammation, edema and blistering in skin. An important mechanism mediating the action of NM and related mustards is oxidative stress. In these studies a modified murine patch-test model was used to analyze DNA damage and the antioxidant/stress response following NM exposure in isolated epidermis. NM (20 μmol) was applied to glass microfiber filters affixed to a shaved dorsal region of skin of CD-1 mice. NM caused structural damage to the stratum corneum as reflected by increases in transepidermal water loss and skin hydration. This was coordinate with edema, mast cell degranulation and epidermal hyperplasia. Within 3 h of NM exposure, a 4-fold increase in phosphorylated histone H2AX, a marker of DNA double-stranded breaks, and a 25-fold increase in phosphorylated p53, a DNA damage marker, were observed in the epidermis. This was associated with a 40% increase in 8-oxo-2'-deoxyguanosine modified DNA in the epidermis and a 4-fold increase in 4-hydroxynonenal modified epidermal proteins. At 12 h post NM, there was a 3-75 fold increase in epidermal expression of antioxidant/stress proteins including heme oxygenase-1, thioredoxin reductase, superoxide dismutase, glutathione reductase, heat shock protein 27 and cyclooxygenase 2. These data indicate that NM induces early oxidative epidermal injury in mouse skin leading to an antioxidant/stress response. Agents that enhance this response may be useful in mitigating mustard-induced skin injury.


Relationship Education for Incarcerated Adults.
As relationship education (RE) programs become more widely implemented, it is important to measure and document the changes associated with RE for diverse audiences. Also, researchers have been challenged to examine the impact of RE with more disadvantaged groups. While we are seeing an increase in this area, only three studies have examined RE with an incarcerated sample. These previous studies examined only those currently in a relationship and focused primarily on couple functioning. The aim of this study was to expand the existing literature by examining RE with a broader sample of incarcerated adults, regardless of current relationship status, and to expand our understanding of its association with outcomes beyond the couple domain by also including measures of individual and parental functioning. In addition, we examined whether change from pre- to posttest was moderated by individual characteristics. Using a sample of incarcerated adults (N = 122), the study found positive change in three domains of functioning (couple, individual, and parental). Specifically, results indicated change on five of the eight outcome variables examined. Overall, we found both similarities and differences among program participants on changes from pre- to posttest. For the majority of outcomes, the positive change from pre- to posttest emerged regardless of individual characteristics.


The Intramedullary Nailing of Adult Femoral Shaft Fracture by the Way of Open Reduction is a Disadvantage or Not?
The purpose of this study is that to evaluate superiority and results of open technique in the treatment of femoral shaft fracture with interlocking intramedullary nailing. The retrospective study is designed to evaluate results of our technique. In this study, the patients that were admitted to the Orthopedics and Traumatology Department of University Hospital that is third level of trauma center. We claim that open technique is not a disadvantage during union process over closed technique in treatment of femoral shaft fracture with interlocking nailing. In this study, 44 patients that were consulted for adult femoral shaft fracture between January 2008 to July 2010 were included. Patients with open fractures, gunshot wounds, neurovascular injuries, and patients that did not have isolated femoral diaphysis fractures were excluded from the study. Clinical and radiological results of the patients were checked periodically. The open interlocking intramedullary nailing was used in treatment. Complete union rate was 90.9 % in 40 patients who were treated with open interlocking intramedullary nailing for adult femoral shaft fracture, and nonunion rate was 9.1 % in four patients. Mean union time was 18.3 weeks (12-36 weeks). Evaluation of 44 patients according to Thoresen criteria was excellent in 22 patients, good in 6 patients, and bad in 4 patients. We think that open technique is an acceptable technique because all results of our study were similar to results of closed intramedullary nailing technique in literature and some advantages of open technique over closed technique.


FVIII inhibitors: pathogenesis and avoidance.
The pathogenesis of inhibitory antibodies has been the focus of major scientific interest over the last decades, and several studies on underlying immune mechanisms and risk factors for formation of these antibodies have been performed with the aim of improving the ability to both predict and prevent their appearance. It seems clear that the decisive factors for the immune response to the deficient factor are multiple and involve components of both a constitutional and therapy-related nature. A scientific concern and obstacle for research in the area of hemophilia is the relatively small cohorts available for studies and the resulting risk of confounded and biased results. Careful interpretation of data is recommended to avoid treatment decisions based on a weak scientific platform. This review will summarize current concepts of the underlying immunological mechanisms and risk factors for development of inhibitory antibodies in patients with hemophilia A and discuss how these findings may be interpreted and influence our clinical management of patients.


Comparison of cow's milk allergy models highlighted higher humoral and Th2 immune responses in BALB/c than C3H/HeNCrl mice.
Cow's milk allergy (CMA) is common in early childhood and the incidence is increasing. However, its mechanisms of action are still not fully understood due to the range of different clinical symptoms. So far, the development of different mouse models has been the best choice to study the molecular mechanisms triggering allergy. However, the selection of suitable strains for the establishment of animal models truly representative of associated human pathologies is still a challenge. Hence, we focused on both C3H/HeNCrl and BALB/c mice to characterize their susceptibility to CMA. After intraperitoneal sensitization, BALB/c and C3H/HeNCrl strains were challenged with β-lactoglobulin (BLG), and compared in allergic symptoms and active immune response, which assessed by specific antibody production and cytokine release. At first, both groups exhibited anaphylaxis, showed specific BLG-related IgE, Th2 response and seemed both suitable for the development of CMA models. However, a detailed analysis revealed that BALB/c had both stronger humoral and Th2 immune responses, producing more antibodies (IgE and IgG/IgG1/IgG2a), and releasing higher levels of Th2-associated cytokines (IL-4, IL-5, IL-13) compared to C3H/HeNCrl mice. Therefore, BALB/c strain would represent a preferential choice in the establishment of CMA models. This study highlights the subtle differences and major outcomes in the selection of mouse strains for the development of suitable food allergy models.


Rational design of a deuterium-containing M2-S31N channel blocker UAWJ280 with in vivo antiviral efficacy against both oseltamivir sensitive and -resistant influenza A viruses.
Seasonal influenza A virus (IAV) infections are among the most important global health problems. FDA-approved antiviral therapies against IAV include neuraminidase inhibitors, M2 inhibitors, and polymerase inhibitor baloxavir. Resistance against adamantanes (amantadine and rimantadine) is widespread as virtually all IAV strains currently circulating in the human population are resistant to adamantanes through the acquisition of the S31N mutation. The neuraminidase inhibitor-resistant strains also contain the M2-S31N mutant, suggesting M2-S31N is a high-profile antiviral drug target. Here we report the development of a novel deuterium-containing M2-S31N inhibitor UAWJ280. UAWJ280 had broad-spectrum antiviral activity against both oseltamivir sensitive and -resistant influenza A strains and had a synergistic antiviral effect in combination with oseltamivir in cell culture. In vivo pharmacokinetic (PK) studies demonstrated that UAWJ280 had favourable PK properties. The in vivo mouse model study showed that UAWJ280 was effective alone or in combination with oseltamivir in improving clinical signs and survival after lethal challenge with an oseltamivir sensitive IAV H1N1 strain. Furthermore, UAWJ280 was also able to ameliorate clinical signs and increase survival when mice were challenged with an oseltamivir-resistant IAV H1N1 strain. In conclusion, we show for the first time that the M2-S31N channel blocker UAWJ280 has in vivo antiviral efficacy in mice that are infected with either oseltamivir sensitive or -resistant IAVs, and it has a synergistic antiviral effect with oseltamivir.


Radiosynthesis and Preclinical Evaluation of 11C-VA426, a Cyclooxygenase-2 Selective Ligand.
Cyclooxygenase-2 (COX-2) is involved in the inflammatory response, and its recurrent overexpression in cancers as well as in neurodegenerative disorders has made it an important target for therapy. For this reason, noninvasive imaging of COX-2 expression may represent an important diagnostic tool. In this work, a COX-2 inhibitor analogue, VA426 [1-(4-fluorophenyl)-3-(2-methoxyethyl)-2-methyl-5-(4-(methylsulfonil)phenyl)-1H-pyrrole], was synthesized and radiolabelled with the 11C radioisotope. The ex vivo biodistribution profile of 11C-VA426 was evaluated in the brain and periphery of healthy rats and mice and in brain and periphery of inflammation models, based on the administration of LPS. 11C-VA426 synthesis with the tBuOK base showed optimal radiochemical yield (15 ± 2%) based on triflate activity, molar activity (range 37-148 GBq/μmol), and radiochemical purity (>95%). Ex vivo biodistribution studies showed a fast uptake of radioactivity but a rapid washout, except in regions expressing COX-2 (lungs, liver, and kidney) both in rats and in mice, with maximum values at 30 and 10 minutes p.i., respectively. LPS administration did not show significant effect on radioactivity accumulation. Celecoxib competition experiments performed in rats and mice treated with LPS produced a general target unrelated reduction of radioactivity concentration in all peripheral tissues and brain areas examined. Finally, in agreement with the negative results obtained from biodistribution experiments, radiometabolites analysis revealed that 11C-VA426 is highly unstable in vivo. This study indicates that the compound 11C-VA426 is not currently suitable to be used as radiopharmaceutical for PET imaging. This family of compounds needs further implementation in order to improve in vivo stability.


Coupled instability of two X-linked genes in Drosophila mauritiana: germinal and somatic mutability.
A highly unstable allele has been isolated at the white locus of Drosophila mauritiana, a sibling species of D. melanogaster. This allele, white-peach (wpch), mutates spontaneously in males and females to give both wild-type and bleached-white derivatives. The mutation frequency is about 10(-3) mutations/generation. There is no evidence for clustering among mutant progeny, and phenotypically wpch flies with mosaic patches of wild-type tissue in the eyes are frequently recovered. Another X-linked locus, plum, is destabilized when wpch is on the same X chromosome.


Elution and characterization of lymphocytes from rheumatoid inflammatory tissue.
Lymphocytes were eluted from the synovial tissue of 19 patients with classical rheumatoid arthritis. The tissue was minced and dissociated by treatment with crude collagenase and DNase. The cell suspension obtained was filtered and incubated in plastic culture flasks overnight at 37 degrees C. The cells that did not adhere to the plastic surface were harvested and the lymphocytes further purified by the Ficoll-Isopaque gradient centrifugation technique. The lymphocyte yield varied from 0.64 to 32 times 10(6) cells. Differential counts showed on the average 85% lymphocytes, 12% mocrophage-like cells, and variable proportions of polymorphonuclear granulocytes, unclassified cells, and dead cells. An average of 77% of the cells were viable as assessed by the trypan blue exclusion test. This cell suspension was investigated for lymphocyte populations. T lymphocytes were predominant in all experiments (mean, 73.6%). The mean percentage of B lymphocytes was 9.7%, whereas the proportion of Fc-receptor-bearing lymphocytes was on the average 6.0%.


Structural and functional organization of germ plasm condensates.
Reproductive success of metazoans relies on germ cells. These cells develop early during embryogenesis, divide and undergo meiosis in the adult to make sperm and oocytes. Unlike somatic cells, germ cells are immortal and transfer their genetic material to new generations. They are also totipotent, as they differentiate into different somatic cell types. The maintenance of immortality and totipotency of germ cells depends on extensive post-transcriptional and post-translational regulation coupled with epigenetic remodeling, processes that begin with the onset of embryogenesis [1, 2]. At the heart of this regulation lie germ granules, membraneless ribonucleoprotein condensates that are specific to the germline cytoplasm called the germ plasm. They are a hallmark of all germ cells and contain several proteins and RNAs that are conserved across species. Interestingly, germ granules are often structured and tend to change through development. In this review, we describe how the structure of germ granules becomes established and discuss possible functional outcomes these structures have during development.


Individual and Neighborhood Predictors of Mental Illness Stigma in New York State.
While studies indicate that stigmatizing attitudes persist in the general public, individual and neighborhood level factors that are associated with increased likelihood of holding stigmatizing attitudes have been seldom studied. This study examined the demographic and neighborhood correlates of stigmatizing attitudes among community members in New York State. Data were drawn from the Pulse of New York State Survey, a random-digit dial survey of 806 New York State residents. Variables studied included demographic information, the Attitudes Toward Mental Illness scale, and neighborhood disadvantage at the zip code level (using data on community characteristics from the 2000 and 2010 Census). Higher levels of completed education predicted less stigmatizing attitudes. Higher levels of neighborhood disadvantage predicted more stigmatizing attitudes with the 2000 Census, and obtained marginal significance within the 2010 Census. Political affiliation demonstrated the strongest relationship, with more conservative ideology predicting more stigmatizing attitudes. Results highlight the need to consider political affiliation and neighborhood disadvantage as target areas when planning interventions for reducing mental illness stigma.


Patients undergoing shoulder arthroplasty for failed nonoperative treatment of proximal humerus fracture have low implant survival and low patient-reported outcomes: 837 cases from the Danish Shoulder Arthroplasty Registry.
Background and purpose - When nonoperative treatment of proximal humerus fracture (PHF) fails, shoulder arthroplasty may be indicated. We investigated risk factors for revision and evaluated patient-reported outcome 1 year after treatment with either stemmed hemiarthroplasty (SHA) or reverse total shoulder arthroplasty (RTSA) after previous nonoperative treatment of PHF sequelae.Patients and methods - Data were derived from the Danish Shoulder Arthroplasty Registry and included 837 shoulder arthroplasties performed for PHF sequelae between 2006 and 2015. Type of arthroplasty, sex, age, and surgery period were investigated as risk factors. The Western Ontario Osteoarthritis of the Shoulder index (WOOS) was used to evaluate patient-reported outcome (0-100, 0 indicates worst outcome). Cox regression and linear regression models were used in the statistical analyses.Results - 644 patients undergoing SHA and 127 patients undergoing RTSA were included. During a mean follow-up of 3.7 years, 48 (7%) SHA and 14 (11%) RTSA were revised. Men undergoing RTSA had a higher revision rate than men undergoing SHA (hazard ratio [HR] 6, 95% confidence interval [CI] 2-19). 454 (62%) patients returned a complete WOOS questionnaire. The mean WOOS score was 53 for SHA and 53 for RTSA. Patients who were 65 years or older had a better WOOS score than younger patients (mean difference 7, CI 1-12). Half of patients had WOOS scores below 50.Interpretation - Shoulder arthroplasty for PHF sequelae was associated with a high risk of revision and a poor patient-reported outcome. Men treated with RTSA had a high risk of revision.


Collimator optimization and collimator-detector response compensation in myocardial perfusion SPECT using the ideal observer with and without model mismatch and an anthropomorphic model observer.
The collimator is the primary factor that determines the spatial resolution and noise tradeoff in myocardial perfusion SPECT images. In this paper, the goal was to find the collimator that optimizes the image quality in terms of a perfusion defect detection task. Since the optimal collimator could depend on the level of approximation of the collimator-detector response (CDR) compensation modeled in reconstruction, we performed this optimization for the cases of modeling the full CDR (including geometric, septal penetration and septal scatter responses), the geometric CDR, or no model of the CDR. We evaluated the performance on the detection task using three model observers. Two observers operated on data in the projection domain: the Ideal Observer (IO) and IO with Model-Mismatch (IO-MM). The third observer was an anthropomorphic Channelized Hotelling Observer (CHO), which operated on reconstructed images. The projection-domain observers have the advantage that they are computationally less intensive. The IO has perfect knowledge of the image formation process, i.e. it has a perfect model of the CDR. The IO-MM takes into account the mismatch between the true (complete and accurate) model and an approximate model, e.g. one that might be used in reconstruction. We evaluated the utility of these projection domain observers in optimizing instrumentation parameters. We investigated a family of 8 parallel-hole collimators, spanning a wide range of resolution and sensitivity tradeoffs, using a population of simulated projection (for the IO and IO-MM) and reconstructed (for the CHO) images that included background variability. We simulated anterolateral and inferior perfusion defects with variable extents and severities. The area under the ROC curve was estimated from the IO, IO-MM, and CHO test statistics and served as the figure-of-merit. The optimal collimator for the IO had a resolution of 9-11 mm FWHM at 10 cm, which is poorer resolution than typical collimators used for MPS. When the IO-MM and CHO used a geometric or no model of the CDR, the optimal collimator shifted toward higher resolution than that obtained using the IO and the CHO with full CDR modeling. With the optimal collimator, the IO-MM and CHO using geometric modeling gave similar performance to full CDR modeling. Collimators with poorer resolution were optimal when CDR modeling was used. The agreement of rankings between the IO-MM and CHO confirmed that the IO-MM is useful for optimization tasks when model mismatch is present due to its substantially reduced computational burden compared to the CHO.


Activation of phenotypically-distinct neuronal subpopulations of the rat amygdala following exposure to predator odor.
Exposure of rats to an odor of a predator can elicit an innate fear response. In addition, such exposure has been shown to activate limbic brain regions such as the amygdala. However, there is a paucity of data on the phenotypic characteristics of the activated amygdalar neurons following predator odor exposure. In the current experiments, rats were exposed to cloth which contained either ferret odor, butyric acid, or no odor for 30 min. Ferret odor-exposed rats displayed an increase in defensive burying versus control rats. Sections of the brains were prepared for dual-labeled immunohistochemistry and counts of c-Fos co-localized with Ca(2+)/calmodulin-dependent protein kinase II (CaMKII), parvalbumin, or calbindin were made in the basolateral (BLA), central (CEA), and medial (MEA) nucleus of the amygdala. Dual-labeled immunohistochemistry showed a significant increase in the percentage of CaMKII-positive neurons also immunoreactive for c-Fos in the BLA, CEA and MEA of ferret odor-exposed rats compared to control and butyric acid-exposed groups. Further results showed a significant decrease in calbindin-immunoreactive neurons that were also c-Fos-positive in the anterior portion of the BLA of ferret odor-exposed rats compared to control and butyric acid-exposed rats, whereas the MEA expressed a significant decrease in calbindin/c-Fos dual-labeled neurons in butyric acid-exposed rats compared to controls and ferret odor-exposed groups. These results enhance our understanding of the functioning of the amygdala following exposure to predator threats by showing phenotypic characteristics of activated amygdalar neurons. With this knowledge, specific neuronal populations could be targeted to further elucidate the fundamental underpinnings of anxiety and could possibly indicate new targets for the therapeutic treatment of anxiety.


The Millennial Mindset
The U.S. Census Bureau defines the Millennial generation as those born between 1982 and 2000, placing the upper age range at 36 years old. The largest generation in US history, outnumbering Baby Boomers and representing more than one-quarter of the US population, they are poised to reshape the economy. They are outspending Baby Boomers 2 to 1 on self-care, and the ASDS 2016 annual survey reported that, in the prior year, patients under 30 saw 20% growth in neuromodulators and 100% growth in injectable filler procedures. As such, they represent a significant segment of the dermatology patient base and understanding how best to communicate with the individuals who comprise this singular collective is key to delivering the best quality and experience of care in a medical or aesthetic practice. A search of PubMed.gov yields no studies relating to the behaviors of millennial patients, and since many of the understandings of the millennial mindset have been developed in the finance, tech, retail, and wellness arenas, we can draw on these insights to enhance connection with the next generation of patients. J Drugs Dermatol. 2018;17(12):1340-1342.


Topical administration of tranexamic acid in total hip arthroplasty: A meta-analysis of Randomized Controlled Trials.
Tranexamic acid (TXA) is an antifibrinolytic drug which has been widely used in many areas of surgery. The purpose of our meta-analysis was to review randomized controlled trials (RCT) of the effectiveness and safety of topical TXA treatment in reducing total blood loss and transfusion rate for patients undergoing primary total hip arthroplasty (THA). A literature search was undertaken. Five eligible reports match the inclusion and exclusion standard. The topical administration of TXA groups revealed lower total blood loss(mean difference - 347 ml, 95% CI, - 411 to - 282 mL; p < 0.00001) and transfusion rate (OR, 0.23 (p < 0.00001; 95% CI, 0.14-0.38)) compared with control groups. Meanwhile there was no statistically significant difference in the risk of developing thromboembolic events (OR, 1.64 (95% CI, 0.39-6.97); p = 0.5). Topical administration of TXA could significantly reduce total blood loss and transfusion requirements in primary THA, without increased thromboembolic complications.


Degradation of mono-oleoylglycerol, trioleoylglycerol and phosphatidylcholine in emulsions and lipoproteins by rat hepatic acylglycerol lipase.
The purpose of this study was to characterize the lipolytic activities released by heparin from rat livers. Heparin perfusates of rat livers degraded monooleoylglycerol, trioleoylglycerol and phosphatidylcholine in emulsions as well as in chylomicrons, chylomicron remnants, low-density lipoprotein/high-density lipoprotein-1 (LDL/HDL-1) and high-density lipoprotein-2 (HDL-2). The preferred substrate was mono-oleoylglycerol. Heparin perfusates were separated by chromatography on either heparin-Sepharose or N-desulphated, N-acetylated heparin-Sepharose into at least two related lipases which differed in their ability to hydrolyse HDL-2 phosphatidylcholine, but not in their ability to degrade mono-oleoylglycerol, trioleoylglycerol and phosphatidylcholine in emulsions. The sodium dodecyl sulphate (SDS)/polyacrylamide-gel-electrophoretic patterns of heparin perfusates purified on either normal or N-desulphated N-acetylated heparin-Sepharose were the same, despite differences in their ability to degrade HDL-2 phosphatidylcholine. There was a single band of Mr 56000 without 2-mercaptoethanol in the SDS disruption buffer and three major bands, of Mr 62000, 59000 and 56000, with 2-mercaptoethanol present. When mono-oleoylglycerol lipase was purified 161-fold, there was a concomitant enrichment of the Mr-56000 protein.


Hyperglycemia impedes lung bacterial clearance in a murine model of cystic fibrosis-related diabetes.
Cystic fibrosis-related diabetes (CFRD) is the most common comorbidity associated with cystic fibrosis (CF), impacting more than half of patients over age 30. CFRD is clinically significant, portending accelerated decline in lung function, more frequent pulmonary exacerbations, and increased mortality. Despite the profound morbidity associated with CFRD, little is known about the underlying CFRD-related pulmonary pathology. Our aim was to develop a murine model of CFRD to explore the hypothesis that elevated glucose in CFRD is associated with reduced lung bacterial clearance. A diabetic phenotype was induced in gut-corrected CF transmembrane conductance regulator (CFTR) knockout mice (CFKO) and their CFTR-expressing wild-type littermates (WT) utilizing streptozotocin. Mice were subsequently challenged with an intratracheal inoculation of Pseudomonas aeruginosa (PAO1) (75 μl of 1-5 × 10(6) cfu/ml) for 18 h. Bronchoalveolar lavage fluid was collected for glucose concentration and cell counts. A portion of the lung was homogenized and cultured as a measure of the remaining viable PAO1 inoculum. Diabetic mice had increased airway glucose compared with nondiabetic mice. The ability to clear bacteria from the lung was significantly reduced in diabetic WT mice and control CFKO mice. Critically, bacterial clearance by diabetic CFKO mice was significantly more diminished compared with nondiabetic CFKO mice, despite an even more robust recruitment of neutrophils to the airways. This finding that CFRD mice boast an exaggerated, but less effective, inflammatory cell response to intratracheal PAO1 challenge presents a novel and useful murine model to help identify therapeutic strategies that promote bacterial clearance in CFRD.


Preliminary study showing the relationship between platelet fibronectin, sialic acid, and ADP-induced aggregation levels in coronary heart disease.
Several studies indicate that thrombosis plays an important role in the pathogenesis of coronary heart disease (CHD). Fibronectin is a multifunctional protein in plasma, other body fluids, and cell surface and plays an important role in platelet functions, including mediation of cell-cell and cell-surface interactions. Sialic acid is a regular constituent of glycoproteins and gangliozides in the outer cell membrane of mammalian cells. Therefore, the sialic acid content of platelets, which are characterized by their ability to aggregate with each other, can be important in leading to thrombus formation. In this study, platelet fibronectin, sialic acid-, and adenosine diphosphate (ADP)-induced platelet aggregation levels were determined in patients with CHD. Platelet sialic acid concentrations were determined by Warren's method. Platelet aggregation tests with ADP in platelet-rich plasma (PRP) were analyzed by use of an aggregometer. Platelet homogenate fibronectin levels were determined by ELISA. Total protein levels were determined by Lowry method. Our results indicate that, in patients with no vessel disease (patients with no obstructed vessel but suffering from chest pain, like angina pectoris) platelet fibronectin levels were significantly lower than the total of the other patients (patients with 1, 2, or 3 obstructed coronary vessels) (p<0.05). Sialic acid levels in patients with no vessel disease were significantly lower than the total of the patient group (p<0.05). There was significant (+) correlation between platelet aggregation, platelet fibronectin, platelet sialic acid, and severity of disease (p<0.05). Our preliminary findings suggest that, especially platelet fibronectin levels potentially represent a pathogenic factor for CHD.


Effects of Hot Packs on Small-Intestinal Motility Measured by Doppler Ultrasound and Subjective Feelings in Normal Adults.
Constipation is a common problem in patients and the general population. Hot packs can relieve constipation, but the effect on small-intestinal motility remains unknown. We aimed to determine the effect of hot packs on small-intestinal motility and subjective feelings associated with bowel activity after removing the hot packs. Thirty-four normal adults were assigned to either an experimental group (n = 18) or a control group (n = 16). Hot and normal packs were applied for 10 minutes to the lumbar regions of the experimental and control groups, respectively. Small-intestinal motility was measured by Doppler ultrasound before, during, and after pack application. Subjective feelings were also evaluated after removing the packs. The number of small-intestinal peristalses and subjective feelings of 20 participants showing anechoic areas in the small-intestinal lumen were analyzed.


Feasibility and performance of cross-clone Raman calibration models in CHO cultivation.
Raman spectroscopy is widely used in monitoring and controlling cell cultivations for biopharmaceutical drug manufacturing. However, its implementation for culture monitoring in the cell line development stage has received little attention. Therefore, the impact of clonal differences, such as productivity and growth, on the prediction accuracy and transferability of Raman calibration models is not yet well described. Raman OPLS models were developed for predicting titer, glucose and lactate using eleven CHO clones from a single cell line. These clones exhibited diverse productivity and growth rates. The calibration models were evaluated for clone-related biases using clone-wise linear regression analysis on cross validated predictions. The results revealed that clonal differences did not affect the prediction of glucose and lactate, but titer models showed a significant clone-related bias, which remained even after applying variable selection methods. The bias was associated with clonal productivity and lead to increased prediction errors when titer models were transferred to cultivations with productivity levels outside the range of their training data. The findings demonstrate the feasibility of Raman-based monitoring of glucose and lactate in cell line development with high accuracy. However, accurate titer prediction requires careful consideration of clonal characteristics during model development.


Spousal homicide and suicide in Quebec.
Domestic violence is a cause for major concern in psychiatry today, yet little is known about the amplitude and dynamics of spousal homicide and extended suicide. Within the jurisdiction of the Quebec Coroner's Office, the investigation files on all consecutive cases of deceased victims of intrafamilial violence occurring between 1991 and 1998 were reviewed. Using a validated checklist, a variety of variables were systematically collected and reviewed for descriptive analysis. Three hundred eighty-eight cases of death were studied. Of this sample, 145 cases (37.4%) concerned victims of conjugal homicide. Fifty-eight cases (40.01%) concerned victims whose homicidal spouses subsequently killed themselves. Suicidal offenders were more likely to be men, to be estranged from their spouse through separation, and, most often, to have used a firearm in the commission of the extended homicide-suicide. The majority of offenders suffered from clinical symptoms of depression. This study emphasizes the need to develop a detailed tool to assist coroners in the field and police investigators with the gathering of specific information that will be of use to clinical researchers.


Cytochrome P450 enzymes in fungal natural product biosynthesis.
Covering: 2015 to the end of 2020 Fungal-derived polyketides, non-ribosomal peptides, terpenoids and their hybrids contribute significantly to the chemical space of total natural products. Cytochrome P450 enzymes play essential roles in fungal natural product biosynthesis with their broad substrate scope, great catalytic versatility and high frequency of involvement. Due to the membrane-bound nature, the functional and mechanistic understandings for fungal P450s have been limited for quite a long time. However, recent technical advances, such as the efficient and precise genome editing techniques and the development of several filamentous fungal strains as heterologous P450 expression hosts, have led to remarkable achievements in fungal P450 studies. Here, we provide a comprehensive review to cover the most recent progresses from 2015 to 2020 on catalytic functions and mechanisms, research methodologies and remaining challenges in the fast-growing field of fungal natural product biosynthetic P450s.


Novel technique for the treatment of stress urinary incontinence. Early experience.
To evaluate the efficacy of in situ anterior vaginal wall sling, reinforced with polypropylene mesh, in the treatment of stress urinary incontinence (SUI). Seventeen patients with mean age of 51.29 years (range: 27-73) underwent surgery for SUI from August 2006 to August 2007 at Osmaniye State Hospital, Osmaniye, Turkey. Two out of 17 patients gave history of previous anti-incontinence surgical intervention, while the remaining were primary cases. In situ anterior vaginal wall sling, reinforced with semi-size monofilament polypropylene tape (size of the mesh was similar to size of in situ sling), was used as an anti-incontinence procedure. The mean follow-up period was 9.29 months (range: 4-16). In-situ anterior vaginal wall sling reinforced with polypropylene mesh was successful in 16 (94.1%) patients, 14 of them were cured, and 2 had clinical improvement. There was no report of preoperative urethral, bladder, or bowel injury. One patient developed postoperative urinary retention that resolved after decreasing the tension of the suspension sutures, and one patient had suprapubic wound sepsis treated by oral antibiotics and anti-inflammatory drugs. No postoperatively significant post-voiding residue was detected. The early results of this technique are encouraging. It is easy to learn, economical, with a good success rate. Urethral erosion is less likely to occur due to the presence of intervening vaginal mucosa. In the future, a prospective study recruiting a larger number of patients undergoing this technique with long-term follow-up is recommended.


Periodic heartbeat reversals cause cardiogenic inspiration and expiration with coupled spiracle leakage in resting blowflies, Calliphora vicina.
Respiration in insects is thought to be independent of the circulatory system because insects typically lack respiratory pigments and because oxygen transport occurs in the gaseous phase through a ramified tracheal system by diffusion and convection directly to the tissues. In the blowfly, as in other insects with periodic heartbeat reversal, the haemolymph is periodically shifted between the anterior body and abdomen, exerting alternating pressure changes on the compliant tracheae in the thorax and in the abdomen. Simultaneous pressure and O2 optode measurements show that, during negative pressure periods, the tracheal partial pressure of oxygen (PO2) increases by 0.5 kPa. In the quiescent fly, tracheal PO2 is rather high (17.5-18.9 kPa), although the thoracic spiracles remain constricted. Microscopic video recordings and reflectance measurements revealed that the dorsal soft edges of the valve lips of the second spiracle leave a very small leak, which is passively widened during backward pulses of the heart. Thus, negative pressure, combined with increased leakage of the spiracle Sp2 valve enable inspiration in the thorax. The positive pressure periods are correlated with a new type of convective CO2 micro-bursts as shown in flow-through measurements. The bulk of the CO2 is, however, released after longer interbursts in macro-bursts with actively opening valves reminiscent of the open phase in a cyclic gas exchange. When the valves open, the PO2 in the thoracic air sacs unexpectedly drops by a mean of 2.75±1.09 kPa, suggesting a displacement of O2 by the transient accumulation of CO2 in the tracheal system before its release.


Effects of age and weaning conditions on blood indicators of oxidative status in pigs.
Weaning is a source of social, nutritional and environmental disorders that challenge piglet health. This study assesses the relevance of using plasma indicators of oxidative status as biomarkers of health around weaning in pigs. Blood antioxidant potential (BAP), hydroperoxides (HPO), oxidative stress index (OSI, e.g. HPO/BAP), vitamin A and E concentrations were investigated in two different trials. Trial A was carried out in an experimental unit to investigate the effects of age (from 12 to 147 days of age), weaning (at 21 or 28 days of age) and management at weaning (in optimal (OC) or deteriorated (DC) conditions) on those parameters. Trial B was performed in 16 commercial pig farms to describe the variability of these indicators on field between 26 and 75 days of age. In trial A, between 12 and 147 days of age, HPO globally increased (P < 0.001), vitamin E concentration decreased (P < 0.001) whereas BAP and vitamin A concentration remained relatively stable (P > 0.1). Vitamins E and A concentrations dropped 5 days after weaning independently of weaning age, weaning conditions and expression of diarrhea (P < 0.001). Twelve days after weaning, whatever the weaning age, HPO and OSI increased in DC compared to OC piglets (P = 0.05 and P < 0.01) and in piglets exhibiting diarrhea compared to those without diarrhea (P < 0.01 and P < 0.001). In DC pigs, BAP was also decreased (P < 0.05) 12 days after weaning. On trial B, plasma concentrations of vitamins A and E decreased and HPO increased 5 and 19 days respectively after weaning (P < 0,001). Contrarily to trial A, BAP values did not drop after weaning. Piglets which had the lowest ADG (Average Daily Gain) after weaning had greater HPO and OSI and lower vitamin A and E concentrations after weaning but also lower vitamin E concentration before weaning (P < 0.05). In conclusion, HPO or OSI seem to be good indicators of health disorders around weaning and plasma concentration of vitamin E before weaning is associated to growth after weaning.


Preparation, characterization, and analysis of multi-walled carbon nanotube-based nanofluid: an aggregate based interpretation.
Nanofluids are gaining attention as an attractive solution for the sustainable machining of difficult-to-cut materials. Despite the enormous recent work in the literature, there are still contradictions concerning the effect of different preparation factors on the characteristics of nanofluids and the underlying mechanisms governing them. In the present study, the effect of varying the preparation factors, namely, multi-walled carbon nanotube (MWCNT) concentration, sonication time, and surfactant amount on various nanofluid characteristics and the interactions among these characteristics were studied. The characteristics are divided into two categories: (a) dispersion/stability and (b) viscosity/wettability. The analysis showed strong interactions between these two categories which were mainly attributed to aggregates' formation and dynamics. For the stability/dispersion responses, the effect of aggregation and saturation phenomena is discussed in relation to the different preparation factors. Our analysis shows that the nanofluid viscosity is strongly dependent on aggregate morphology. As for wettability, a novel mechanism is proposed and used to explain the nanoparticles' influence on wettability based on the nanolayering theory. Finally, multi objective optimization (MOO) based on grey relational analysis (GRA) was performed. It was found that moderate MWCNT concentration, high sonication time, and low surfactant amount show the optimal characteristics within the current study design variables search domain. The novelty in the present study lies in its consideration of the simultaneous interaction between the nanofluids' properties and stability. Unlike the common practice in the literature, which focuses on one or two aspects of nanofluids, our approach broadens the analysis and provides in-depth insights into the nanofluid as a complete physical system.


Myelin instability and oligodendrocyte metabolism in myelin-deficient mutant mice.
During the active phase of myelination in myelin-deficient mutant mice (mld), myelin basic protein (MBP) synthesis is defective and the myelin lamellae are uncompacted. In these mutants, we found a fast metabolism of the myelin-associated glycoprotein (MAG) and of sulfatides, and the presence of cholesterol esters and a degradation product of MAG, dMAG, indicating that mld myelin was unstable. The increased synthesis of MAG and Wolfgram protein, two proteins present in uncompacted myelin sheath and paranodal loops, was demonstrated by high levels of messengers. Simultaneously, we found an accumulation of inclusion bodies, vacuoles, and rough endoplasmic reticulum in mld oligodendrocytes. This material was heavily immunostained for MAG. Furthermore, the developmental change between the two molecular forms of MAG (p72MAG/p67MAG) was delayed in mld mice. In 85-d-old mld mice, the MBP content increased and myelin lamellae became better compacted. In these mutants, dMAG was absent and MAG mRNAs were found in normal amounts. Furthermore, the fine structure of mld oligodendrocytes was normal and the MAG immunostaining was similar to age-matched controls. These results support a functional role for MBP in maintaining the metabolic stability and the compact structure of myelin. Furthermore, in the absence of MBP and myelin compaction, the regulation of the synthesis of at least two membrane proteins related to myelin cannot proceed.


Primetime learning: collaborative and technology-enhanced studying with genuine teacher presence.
Productive learning processes and good learning outcomes can be attained by applying the basic elements of active learning. The basic elements include fostering discussions and disputations, facing alternative conceptions, and focusing on conceptual understanding. However, in the face of poor course retention and high dropout rates, even learning outcomes can become of secondary importance. To address these challenges, we developed a research-based instructional strategy, the primetime learning model. We devised the model by organizing the basic elements of active learning into a theory-based four-step study process. The model is based on collaborative and technology-enhanced learning, on versatile formative assessment without a final exam, and on genuine teacher presence through intimate meetings between students and teachers. We piloted the model in two university physics courses on thermodynamics and optics and observed persistent student activity, improved retention, and good learning outcomes. Feedback suggested that most students were satisfied with the learning experience. The model suits particularly well for courses that, in addition to the teaching subject itself, focus on teaching balanced study habits and strengthening social integration. By its very construction, it also helps the propagation of research-based instructional strategies. Although the model does contain challenges, it represents a generic framework for learning and teaching that is flexible for further development and applicable to many subjects and levels.


Salter-Harris II fracture of the proximal humerus in a preterm infant.
A premature twin infant presented with clinical signs of shoulder injury shortly after birth. He had been delivered by cesarean section due to malpresentation. Bony abnormality was not initially recognized on plain radiographs, but ultrasound revealed a Salter-Harris II fracture of the proximal humerus. Plain radiographs are limited in the premature newborn because of lack of calcification of the epiphysis. Ultrasound examination in these infants can be helpful in determining the true extent of skeletal injury. Immobilization and pain control are then indicated to optimize the outcome.


Automated crystal system identification from electron diffraction patterns using multiview opinion fusion machine learning.
A bottleneck in high-throughput nanomaterials discovery is the pace at which new materials can be structurally characterized. Although current machine learning (ML) methods show promise for the automated processing of electron diffraction patterns (DPs), they fail in high-throughput experiments where DPs are collected from crystals with random orientations. Inspired by the human decision-making process, a framework for automated crystal system classification from DPs with arbitrary orientations was developed. A convolutional neural network was trained using evidential deep learning, and the predictive uncertainties were quantified and leveraged to fuse multiview predictions. Using vector map representations of DPs, the framework achieves a testing accuracy of 0.94 in the examples considered, is robust to noise, and retains remarkable accuracy using experimental data. This work highlights the ability of ML to be used to accelerate experimental high-throughput materials data analytics.


Calcitonin-induced change in serum calcium levels and its relationship to osteoclast morphology and number of calcitonin receptors.
It has been shown that, in live subjects, the ability of calcitonin (CT) to decrease serum calcium (Ca) levels can be lost in response to its continued or repeated administration. The present study investigated the relationship between such changes of in vivo serum Ca levels and the response of osteoclasts to CT administration, including the downregulation of their CT receptors (CTRs). Rats were either given a single injection of CT or repeated injections at either 6- or 24-h intervals, after which their serum Ca levels were evaluated. Their parietal bones were dissected, and the amount of 125I-labeled elcatonin (125I-eCT) binding to their osteoclasts measured using autoradiography. Ultrastructural changes in the osteoclasts were also examined. Twenty-four hours after a single CT administration, serum Ca levels had dropped, and there was an absence of ruffled borders on the osteoclasts. Less 125I-eCT binding to the osteoclast was found than in the control group. Forty-eight and 72 h after CT administration, serum Ca levels had almost returned to control levels, and the osteoclasts showed ruffled borders once again. The amount of 125I-eCT binding to the osteoclast also recovered to control levels. When these osteoclasts were then incubated in CT, their ruffled borders once again disappeared. In the 6-h interval multiple CT administration schedule subjects, upon inspection 72 h after their first administration (6 h following the final one), serum Ca levels were found to have almost returned to control levels with the presence of osteoclast ruffled borders. The amount of 125I-eCT binding to these osteoclasts was remarkably limited, and no disappearance of the ruffled borders occurred in response to additional CT incubation. In the 24-h interval multiple administration schedule subjects, upon inspection 72 h after their first CT administration (24 h following the final one), there was less 125I-eCT binding than in the single-dose subjects tested 24 h after their injection, and the ability of CT to lower their serum Ca levels was reduced. The ability of CT to lower serum Ca levels was therefore related to the response of osteoclasts to the CT (the disappearance of the ruffled borders), and this response was related to the amount of CTRs available for binding with CT on the osteoclast surface. Furthermore, the reduced effectiveness of CT in response to repeated CT administration was found to be related to the downregulation of the CTRs on the osteoclast surface.


Systematically expanding nondirect product bases within the pruned multi-configuration time-dependent Hartree (MCTDH) method: A comparison with multi-layer MCTDH.
We propose a pruned multi-configuration time-dependent Hartree (MCTDH) method with systematically expanding nondirect product bases and use it to solve the time-independent Schrödinger equation. No pre-determined pruning condition is required to select the basis functions. Using about 65 000 basis functions, we calculate the first 69 vibrational eigenpairs of acetonitrile, CH3CN, to an accuracy better than that achieved in a previous pruned MCTDH calculation which required more than 100 000 basis functions. In addition, we compare the new pruned MCTDH method with the established multi-layer MCTDH (ML-MCTDH) scheme and determine that although ML-MCTDH is somewhat more efficient when low or intermediate accuracy is desired, pruned MCTDH is more efficient when high accuracy is required. In our largest calculation, the vast majority of the energies have errors smaller than 0.01 cm-1.


Classification of Actinobacillus spp isolates from horses involved in mare reproductive loss syndrome.
To identify Actinobacillus spp isolates recovered from fetuses and pericardial fluid from horses affected with mare reproductive loss syndrome (MRLS) and determine whether these bacterial species are the same as those isolated from clinically normal horses. Isolates of actinobacilli recovered from 18 horses with pericarditis and 109 fetuses aborted by mares affected by MRLS. Procedures-Actinobacillus spp isolates were identified to the level of species or subspecies by use of conventional phenotypic tests and biochemical and enzyme test kits. The 16S rRNA gene from selected isolates was amplified, purified, and sequenced. Sequence data were compared with sequence data for actinobacilli in GenBank. Of the 109 isolates obtained from fetuses, 14 were Actinobacillus equuli subsp equuli, 65 were A equuli subsp haemolyticus, 28 were Bisgaard taxon 10-like bacterium, and 2 were Actinobacillus genomospecies 1. Of the 18 isolates from horses with pericarditis, 4 were A equuli subsp equuli, 13 were A equuli subsp haemolyticus, and 1 was Bisgaard taxon 10-like bacterium. Comparisons with published data and GenBank data revealed that the isolates recovered from horses with MRLS were the same as those isolated from the oral cavity or alimentary tract of healthy horses. Actinobacillus spp isolates recovered from fetuses and pericardial fluid samples of horses affected by MRLS in 2001 to 2003 were identical to Actinobacillus spp found in the oral cavity and alimentary tracts of healthy horses.


The posterior thalamic region and its cortical projection in New World and Old World monkeys.
The posterior nuclear complex of the thalamus in rhesus, pigtailed and squirrel monkeys consists of the combined suprageniculate-limitans nucleus and an ill defined region of heterogeneous cell types extending anteriorly from the dorsal lobe of the medial geniculate body towards the posterior pole of the ventral nuclear complex. This region is referred to as the posterior nucleus. It is directly continuous with the ventroposteroinferior nucleus. The cortical projections of each of these nuclei, together with those of the adjacent ventral, pulvinar and medial geniculate complexes, have been studied by means of the autoradiographic tracing technique. The suprageniculate-limitans nucleus, the main input to which is the superior colliculus, projects upon the granular insular area of the cortex. The medial portion of the posterior nucleus projects to the retroinsular field lying posterior to the second somatic sensory area. There is clinical and electrophysiological evidence to suggest that the retroinsular area may form part of a central pain pathway. The lateral portion of the posterior nucleus which is closely related to certain elements of the medial geniculate complex, projects to the postauditory cortical field. The ventroposterioinferior nucleus, which may be involved in vestibular function, projects to the dysgranular insular field. The principal medial geniculate nucleus can be subdivided into a ventral division that projects to field AI of the auditory cortex and a dorsal division that merges with the posterior nucleus; it is further subdivided into an anterodorsal component that projects to two fields on the superior temporal gyrus, together with a posterodorsal component in which separate cell populations project to areas lying anterior and medial to AI. The magnocellular medial geniculate nucleus, sometimes considered a part of the posterior complex, appears to project diffusely to layer I of all the auditory fields. The auditory fields are bounded on three sides by the projection field of the medial nucleus of the pulvinar which also extends into the upper end of the lateral sulcus to bound the fields receiving fibers from the posterior nucleus. The topography of the areas receiving fibers from the posterior, medial geniculate and pulvinar complexes, taken in conjunction with the rotation of the primate temporal lobe, permits all of these fields to be compared with similar, better known areas in the cat brain.


Assessment of renal function in routine care of people living with HIV on ART in a resource-limited setting in urban Zambia.
Data on renal impairment in sub-Saharan Africa (SSA) remains scarce, determination of renal function is not part of routine assessments. We evaluated renal function and blood pressure in a cohort of people living with HIV (PLWH) on antiretroviral treatment (ART) in the Renal Care Zambia project (ReCaZa). Using routine data from an HIV outpatient clinic from 2011-2013, we retrospectively estimated the glomerular filtration rate (eGFR, CKD-Epi formula) of PLWH on ART in Lusaka, Zambia. Data were included if adults had had at least one serum creatinine recorded and had been on ART for a minimum of three months. We investigated the differences in eGFR between ART subgroups with and without tenofovir disproxil fumarate (TDF), and applied multivariable linear models to associate ART and eGFR, adjusted for eGFR before ART initiation. Among 1118 PLWH (63,3% female, mean age 41.8 years, 83% ever on TDF; median duration 1461 [range 98 to 4342] days) on ART, 28.3% had an eGFR <90 ml/min, and 5.5% <60 ml/min at their last measurement. Information on other conditions associated with renal impairment was not systematically documented. Fourteen per cent of the PLWH who later switched to TDF-free ART had an initial eGFR lower 60ml/min. Nineteen percent had first-time hypertensive readings at their last visit. The multivariable models suggest that physicians acted according to guidelines and replaced TDF-containing ART if patients developed moderate/severe renal impairment. Assessment of renal function in SSA remains a challenge. The vast majority of PLWH benefit from long-term ART, including improved renal function. However, approximately 5% of PLWH on ART may have clinically relevant decreased eGFR, and 27% hypertension. While a routine renal assessment might not be feasible, strategies to identify patients at risk are warranted. Targeted monitoring prior and during ART is recommended, however, should not delay ART access.