The next generation of DNA profiling--STR typing by multiplexed PCR--ion-pair RP LC-ESI time-of-flight MS.
For the first time a multiplexed PCR approach suitable for mass spectrometric STR allele identification is presented. Thirteen forensically important STR markers (vWA, D21S11, D3S1358, D16S539, D8S1179, D7S820, D13S317, D5S818, TPOX, CSF1PO, D2S441, D10S1248, and D22S1045) and the gender typing locus amelogenin were simultaneously amplified. Ion-pair reversed-phase high-performance liquid chromatography electrospray-ionization time-of-flight mass spectrometry (ICEMS) was applied for genotyping, and allowed for highly efficient characterization of multiple PCR amplicons. Compared with electrophoretic sizing ICEMS enabled for the simultaneous detection of length and nucleotide variations. Thus, the obtained amount of biological information present within STR profiles was significantly increased even though the compatibility of typing results with electrophoretically generated data(bases) was maintained. Other advantages of the ICEMS platform included the abandonment of internal size standards, allelic ladders, and any kind of spectral calibration. The 14-plex PCR was tailor-made for ICEMS analysis by designing primer pairs that bind close to the repeat region, by using a proof reading polymerase for amplification, and by implementing molecular mass modifiers for prevention of molecular mass overlaps. In a series of experiments, the performance of the multiplexed PCR-ICEMS assay was evaluated. The ICEMS-based DNA profiling assay was found to be competitive regarding detection sensitivity and analyzability of degraded and casework samples with commercially available electrophoretic typing approaches, which suggests that multiplexed PCR-ICEMS assays could represent a valuable tool for (forensic) genetics.


Cardiovascular effects of anandamide in anesthetized and conscious normotensive and hypertensive rats.
We previously showed that in anesthetized rats anandamide elicits bradycardia and a triphasic blood pressure response: transient hypotension secondary to a vagally mediated bradycardia, followed by a brief pressor and prolonged depressor response, the latter two effects being similar to those of delta 9-tetrahydrocannabinol (THC). The prolonged depressor but not the pressor response was reduced after alpha-adrenergic receptor blockade or cervical spinal cord transection and was inhibited by the cannabinoid type 1 (CB1) receptor antagonist SR141716A, suggesting CB1 receptor-mediated sympathoinhibition as the underlying mechanism. Here we examined the relationship between sympathetic tone and the cardiovascular effects of anandamide by testing these effects in both conscious and anesthetized, normotensive and spontaneously hypertensive rats. In urethane-anesthetized normotensive rats, SR141716A inhibited the prolonged depressor and bradycardic effects of anandamide and THC with similar potency, whereas it did not affect the pressor response to either agent. Anadamide caused similar hypotension in spontaneously breathing and in paralyzed, mechanically ventilated rats, suggesting that the hypotension is not secondary to respiratory effects. In conscious normotensive rats, anandamide elicited transient vagal activation and a brief pressor response, but the prolonged hypotensive component was absent. SR141716A potentiated and prolonged the brief pressor response to anandamide, suggesting that the depressor response may have been masked by an increased pressor response. All three phases of the anadamide response were present in both anesthetized and conscious spontaneously hypertensive rats, and the hypotensive component, inhibited by SR141716A in both, was more prolonged in the absence (> 50 minutes) than the presence (10 to 15 minutes) of anesthesia. We conclude that anandamide causes a non-CB1 receptor-mediated pressor and a CB1 receptor-mediated prolonged depressor response. The depressor response can be elicited in both conscious and anesthetized animals, but its magnitude depends on preexisting sympathetic tone.


Social anxiety and HIV transmission risk among HIV-seropositive male patients.
The role of psychological factors in predicting HIV sexual transmission risk behavior is increasingly of interest in prevention research. Social anxiety, or anxiety about being evaluated in interpersonal situations, is associated with unprotected insertive anal intercourse among young men who have sex with men (MSM) and with other behavioral risk factors for unprotected intercourse, such as depression, smoking, alcohol use, and drug use. Social anxiety may be especially relevant in understanding HIV risk among HIV-seropositive men, given its stronger association with unprotected insertive than with receptive anal intercourse. In the present study, for which participants were recruited between October 2002 and May 2003, HIV-positive men attending regularly scheduled primary care medical appointments at a community HIV clinic were approached by research personnel and informed about the study topic and procedures. Ninety percent of patients approached agreed to participate, resulting in a sample of 206 patients. The sample was primarily African American, unemployed, of low educational level, and 95% of the sample had an AIDS diagnosis. The present study replicated and extended previous research from community samples by demonstrating an association between social anxiety and unprotected insertive anal intercourse with non-HIV-positive partners in a clinical sample of HIV-positive MSM and men who have sex with women (MSW). This association was maintained controlling for depression, smoking, and club drug use. Social anxiety is a relatively robust risk factor for unprotected insertive anal intercourse among MSM. Future work should examine the mechanisms by which social anxiety is associated with sexual risk among MSM.


Identification of an oligodeoxynucleotide sequence motif that specifically inhibits phosphorylation by protein tyrosine kinases.
Protein tyrosine kinases (PTKs) have central roles in cellular signal transduction. We have identified a sequence motif (CGT[C]GA) in phosphorothioate-modified oligodeoxynucleotides (ODNs) that specifically inhibits the enzymatic activity of recombinant or immunoprecipitated PTK in vitro. Hexamer ODNs containing this motif block both substrate and autophosphorylation of at least four different PTKs but have no apparent effect on the enzymatic activity of a serine/threonine protein kinase. These data suggest possible new applications for ODNs and have implications for the design and interpretation of experiments using antisense or triplex ODNs.


Attitudes toward straight, gay male, and transsexual parenting.
This study examined American attitudes toward transsexual and gay male parenting, compared to straight parenting. After reporting levels of transphobia, participants read a vignette regarding a couple seeking child adoption. Individuals high in transphobia perceived nontraditional couples as more emotionally unstable than straight couples and were less willing to grant custody of a child to the nontraditional couples vs. the straight couples. In addition, the transsexual couple faced more prejudice and discrimination than the gay male couple. Implications of these findings are discussed.


Involvement of oxyradicals in promotion/inhibition of expansion growth in cucumber cotyledons.
Cytokinin-induced expansion growth of cucumber cotyledons and its interaction with UV-B (280-320 nm) was studied with reference to oxyradicals. UV-B radiation enhanced the level of oxyradicals in the cotyledons measured by EPR spectroscopy. Cytokinin promoted expansion growth was inhibited by UV-B radiation. Cytokinins reduced the level of oxyradicals in dark grown cotyledons, while promoting growth. Overproduction of oxyradicals by UV-B could not be fully accounted for the inhibition of growth of cotyledons, since quenching of radicals by cytokinins did not fully restore inhibition of growth.


Iterative linearized approach to nonadiabatic dynamics.
This paper presents a new approach to propagating the density matrix based on a time stepping procedure arising from a Trotter factorization and combining the forward and backward incremental propagators. The sums over intermediate states of the discrete quantum subsystem are implemented by a Monte Carlo surface hopping-like procedure, while the integrals over the continuous variables are performed using a linearization in the difference between the forward and backward paths of these variables leading to classical-like equations of motion with forces determined by the quantum subsystem states. The approach is tested on several models and numerical convergence is explored.


Equivalence of a one-dimensional driven-diffusive system and an equilibrium two-dimensional walk model.
It is known that a single product shock measure in some of the one-dimensional driven-diffusive systems with nearest-neighbor interactions might evolve in time quite similarly to a random walker moving on a one-dimensional lattice with reflecting boundaries. The nonequilibrium steady state of the system in this case can be written in terms of a linear superposition of such uncorrelated shocks. Equivalently, one can write the steady state of this system using a matrix-product approach with two-dimensional matrices. In this paper, we present an equilibrium two-dimensional one-transit walk model and find its partition function using a transfer matrix method. We will show that there is a direct connection between the partition functions of these two systems. We will explicitly show that in the steady state, the transfer matrix of the one-transit walk model is related to the matrix representation of the algebra of the driven-diffusive model through a similarity transformation. The physical quantities are also related through the same transformation.


Closure of the interventricular foramen and morphogenesis of the membranous septum and ventricular septal defects in the human heart.
The development of the atrioventricular and interventricular membranous septum was studied The interventricular foramen is closed by the growth of the endocardial crest of the interventricular muscular septum towards the cranio-ventral atrioventricular cushion and towards the part B of the outflow tract ridge B-3; the bulbus is septated by the apposition and fusion of parts A and B of the outflow tract ridges A-1 and B-3. The membranous septum develops from the crest of the interventricular muscular septum between the interventricular muscular septum and the right border of the atrioventricular cushions, where it forms the atrioventricular mesenchymal septum. This last develops towards the right surface of the interventricular muscular septum and forms the septal cusp primordium of he tricuspid valve; this is undermined, for which reason the septal cusp is cut off from the interventricular muscular septum at the 5th month of fetal life. From the 6th month of intrauterine life, the undermining extends to the atrioventricular mesenchymal septum, for which reason the right septal cusp becomes inserted in the right face of the atrioventricular mesenchymal septum which becomes divided into atrioventricular and interventricular. The Authors advance some hypotheses as to the origin of certain defects of the interventricular septum.


[Interleukin 2 secretion by lymphocytes of patients with squamous cell cancer of the oropharynx or larynx].
The various strengthening influences of interleukin-2 (IL-2) on the immunological system is of intrinsic importance to tumour defense. IL-2 concentration can be measured by the determination of the IL-2 dependent CTLL cell line in a micro-Elisa-assay. The IL-2 secretion is dose dependent after stimulation with the mitogens phytohaemagglutinin A (PHA), phorbolester (PMA) or the monoclonal T cell antibody OKT3. Maximal IL-2 release is achieved by concomitant activation with phytohaemagglutinin A (PHA) and phorbolester (PMA). As controls healthy donors with low and high alcohol and cigarette consumption were used. In patients with cancer of the oropharynx, a significant reduction in the IL-2 secretion was observed when compared with the larynx cancer group or with healthy donors. This might suppose an immunological influence of the squamous cell carcinoma which grows in the oropharynx in close proximity to lymphatic cells and vessels which are numerously present in the tonsillar ring of Waldeyer.


Treatment of periodontal disease in a patient with Ehlers-Danlos syndrome. A case report and literature review.
Ehlers-Danlos syndrome (EDS) designates a heterogeneous group of connective tissue disorders characterized by skin elasticity, tissue fragility, and chronic joint pain. Dental findings have been reported with some types of EDS. This case report describes the periodontal findings in a patient with a previously undiagnosed EDS type VIII. Diagnostic aids utilized included microbial testing, histological examination, gingival crevicular fluid (GCF) analysis for the levels of C-telopeptide pyridinoline cross-links (ICTP), and genetic counseling. Periodontal treatment consisted of mechanical debridement and adjunctive antibiotic therapy. Genetic counseling and clinical presentation confirmed the diagnosis of EDS type VIII. Periodontal treatment led to marked clinical improvements and GCF levels of the bone resorptive marker ICTP were significantly reduced. The patient and her siblings are currently pursuing appropriate medical care and genetic counseling. Periodontal involvement may lead to the diagnosis of an underlying systemic condition. Identification of suspected etiological factors of periodontal disease may prove critical for the general well-being of some patients.


Spatial patterns in prostate Cancer-specific mortality in Pennsylvania using Pennsylvania Cancer registry data, 2004-2014.
Spatial heterogeneity of prostate cancer-specific mortality in Pennsylvania remains unclear. We utilized advanced geospatial survival regressions to examine spatial variation of prostate cancer-specific mortality in PA and evaluate potential effects of individual- and county-level risk factors. Prostate cancer cases, aged ≥40 years, were identified in the 2004-2014 Pennsylvania Cancer Registry. The 2018 County Health Rankings data and the 2014 U.S. Environmental Protection Agency's Environmental Quality Index were used to extract county-level data. The accelerated failure time models with spatial frailties for geographical correlations were used to assess prostate cancer-specific mortality rates for Pennsylvania and by the Penn State Cancer Institute (PSCI) 28-county catchment area. Secondary assessment based on estimated spatial frailties was conducted to identify potential health and environmental risk factors for mortality. There were 94,274 cases included. The 5-year survival rate in PA was 82% (95% confidence interval, CI: 81.1-82.8%), with the catchment area having a lower survival rate 81% (95% CI: 79.5-82.6%) compared to the non-catchment area rate of 82.3% (95% CI: 81.4-83.2%). Black men, uninsured, more aggressive prostate cancer, rural and urban Appalachia, positive lymph nodes, and no definitive treatment were associated with lower survival. Several county-level health (i.e., poor physical activity) and environmental factors in air and land (i.e., defoliate chemical applied) were associated with higher mortality rates. Spatial variations in prostate cancer-specific mortality rates exist in Pennsylvania with a higher risk in the PSCI's catchment area, in particular, rural-Appalachia. County-level health and environmental factors may contribute to spatial heterogeneity in prostate cancer-specific mortality.


Weight Gain During Treatment of Bipolar Disorder (BD)-Facts and Therapeutic Options.
Bipolar disorder (BPD) is a mood disorder, which is characterized by alternating affective states, namely (hypo)mania, depression, and euthymia. Evidence is growing that BPD has indeed a biologic substrate characterized by chronic inflammation, oxidative stress, and disturbed energy metabolism. Apart from this, there is obviously a hereditary component of this disease with multi-genetic factors. Most probably a susceptibility threshold favors the outbreak of clinical disease after a cascade of stress events that remain to be elucidated in more detail. Evidence is also growing that weak points in brain energy metabolism contribute to outbreak and severity of BPD. Conventional psychopharmacologic therapy must be reassessed under the aspects of weight cycling and development of central obesity as a deterioration factor for a worse clinical course leading to early cardiovascular events in BPD subgroups.


[Effect of NLRP3 mediated pyroptosis in myocardial cells undergoing hypoxia/deoxygenation injury].
Objective: To investigate the effect of NACHT-LRR-PYD- containing proteins 3 (NLRP3) mediated pyroptosis in myocardial cells undergoing hypoxia/deoxygenation (H/R) injury. Methods: In order to observe whether H/R-treatment could cause pyroptosis, H9c2 cells were divided into 2 groups randomly using the lottery method: control group(without H/R-treatment) and H/R group (in which the H9c2 cells were underwent H/R-treatment). In order to clarify the role of pyroptosis in H/R-injury, H9c2 cells were divided into 4 groups randomly using the lottery method: control group(in which the H9c2 cells were cultivated with normal medium); YVAD group(in which the H9c2 cells were pretreated with z-Val-Ala-Asp(Ome)-fluoromethylketone (Z-YVAD-FMK) 20 μm for 4 hours, then replaced with normal medium); H/R group(H9c2 cells underwent H/R-treatment); YVAD+H/R group (in which the H9c2 cells were pretreated with 20 μm Z-YVAD-FMK for 4 hours before H/R-treatment). To determine whether H/R-induced cell pyroptosis is associated with NLRP3, H9c2 cells were divided into 4 groups randomly using the lottery method: control group (in which cells were transfected with a control nonspecific siRNA); si-NLRP3 group (in which cells were transfected with NLRP3-targeting siRNA); H/R group(in which cells were transfected with a control nonspecific siRNA before H/R-treatment); si-NLRP3+H/R group(in which the H9c2 cells were transfected with NLRP3-targeting siRNA before H/R-treatment). Pore formation on cell membrane was detected by propidium iodide (PI) staining. Cell viability was detected by CCK8 reagent. The protein expression of Caspase-1, interleukin-1β (IL-1β) and NLRP3 was detected by Western blot. Results: (1) The positive rate of PI staining ((26.46±5.15)% vs. (1.69±0.73)%,P<0.01), expression of NLRP3 (0.57±0.16 vs. 0.23±0.06,P<0.01), expression of Caspase-1 (1.07±0.13 vs. 0.37±0.08,P<0.01), and expression of IL-1β (0.38±0.08 vs. 0.16±0.05,P<0.01) were significantly higher in H/R group than in control group. (2)The cell vitality was significantly higher in YVAD+H/R group than in H/R group ((87.31±9.05)% vs. (73.30±7.19)%, P<0.05).The positive rate of PI staining was significantly decreased in YVAD+H/R group than in H/R group ((18.12±4.36)% vs. (26.45±4.60)%, P<0.05). The expression of Caspase-1 (0.72±0.12 vs. 1.07±0.15, P<0.05) and IL-1β(0.29±0.07 vs. 0.39±0.06, P<0.05) were significantly lower in YVAD+H/R group than in H/R group. (3) The cell vitality was significantly increased in si-NLRP3+H/R group than in H/R group ((85.46±7.71)% vs. (72.41±5.53)%, P<0.05). The positive rate of PI staining was significantly lower in si-NLRP3+H/R group than in H/R group ((18.22±4.20)% vs. (26.73±3.26)%, P<0.05). The expression of Caspase-1(0.87±0.07 vs. 1.15±0.15, P<0.05) and IL-1β(0.41±0.07 vs. 0.58±0.10, P<0.05) were significantly decreased in si-NLRP3+H/R group than in H/R group. Conclusion: NLRP3 mediated pyroptosis is involved in H/R injury of myocardial cells.


Mitochondria-independent induction of Fas-mediated apoptosis by MSSP.
Fas-mediated apoptosis has been proposed to play an important role in homeostasis. Fas triggers apoptosis after stimulation by its ligand FasL or the Fas ligand agonist anti-Fas antibody through a mitochondria-dependent or -independent pathway, and MSSP has been identified as a transcription factor that regulates the c-myc gene and was later found to positively or negatively regulate a variety of genes, including alpha-smooth actin, MHC class I, MHC class 2 and the thyrotropin receptor. We further found that expression of the Fas gene was repressed, resulting in abrogation of the Fas-mediated induction of apoptosis both in Mssp-knockout mice and primary thymocytes. MSSP was then found to stimulate promoter activity of the Fas gene by binding to a specific region. In this study, to identify the MSSP-dependent Fas-induced apoptosis pathway, primary fibroblasts from MSSP (+/+) and MSSP (-/-) cells were treated with the combination of interleukin 1-beta and interferon-gamma and expression of the Fas gene was examined. The results showed that the Fas gene was expressed at the same levels in the two cell types. Furthermore, when these cells were treated with the anti-Fas antibody, it was found that cytochrome C was not released in the cytosol and that activations of caspase 8 and caspase 3 occurred in primary fibroblasts from MSSP (+/+) cells but not from MSSP (-/-) cells. These results indicate that Fas-mediated apoptosis induced by MSSP occurs independently of mitochondria.


Clinical profile of 300 men with facial hypermelanosis.
Facial hypermelanosis is a significant cause of cosmetic disfigurement, social embarrassment and psychological morbidity affecting quality of life. To study clinicoepidemlogic patterns of facial hypermelanoses among men. Medical records of all adult males presenting with facial hypermelanoses were analyzed for this retrospective cross sectional study for demographic details, duration, cosmetic usage, sun exposure, drug intake, infections, systemic or cutaneous diseases, and family history of hypermelanotic dermatosis. Laboratory investigations and skin biopsy were performed when deemed necessary. These were 300 Indian men aged 18 to 74 (mean 37.35) years with 121 (40.3%) individuals aged 31-50 years. Various patterns of melasma in 230 (76.7%) patients were the major cause of facial hypermelanosis. Periorbital hypermelanosis was observed in 32 (10.7%), freckles and lentigens in 26 (8.7%), acanthosis nigricans in 12 (4%) and lichen planus pigmentosus in 10 (3.3%), pigmented cosmetic contact dermatitis in 7, and nevus of Ota in 6 persons. The 71 (30.8%) patients with melasma had a history of frequent sun exposure, 9 (3.9%) patients had systemic comorbidities. Family history of periorbital melanosis was present in 7 (21.8%), personal or family history of atopy in 5 (15.6%) patients. Acanthosis nigricans was associated with obesity in 9 (75%) of patients and with diabetes mellitus in 4 (33.3%) cases. Melasma, periorbital hypermelanosis, acanthosis nigricans and lichen planus pigmentosus remain the predominant causes for facial hypermelanosis in men.


Assembly of the tendon extracellular matrix during development.
The assembly of the collagenous extracellular matrix during tendon development was studied to determine the mechanisms involved in collagen fibril growth. Developing avian metatarsal tendons were studied using structural, immunochemical and biochemical approaches. Tendon fibroblasts were shown to establish a hierarchy of extracellular compartments associated with fibrils, bundles, and macroaggregates during development. These distinct domains provide a mechanism for the fibroblast to influence the extracellular steps in matrix assembly. A discontinuous fibrillar matrix was assembled and fibril segments approximately 10-20 microns long were deposited into bundles by the 14 d embryo fibroblasts. The fibril segment is a normal assembly intermediate, permitting orderly linear, lateral and intercalatory development and growth. A lateral and/or linear fusion of segments may be responsible for the formation of mature continuous fibrils. Fibril segments were isolated from 12-18 d chick embryo metatarsal tendons. Homogenisation almost completely disrupted the 12-15 d tendons. Transmission electron microscopy demonstrated intact segments. Between d 12 and 15 of development, mean segment length increased from approximately 22 to 33 microns. The incremental increase in length with development indicates a limited linear fusion of segments which is supported by morphological examination. At 16 d, there was a significant decrease in segment extractability and by 17-18 d, intact segments were unextractable. Mean segment lengths were 37 microns and > 70 microns for 16 and 17 d tendons respectively. During this period, fibril diameter also increased.(ABSTRACT TRUNCATED AT 250 WORDS)


Arachidonic acid potentiates exocytosis and allows neuronal SNARE complex to interact with Munc18a.
Neuronal communication relies on the fusion of neurotransmitter-containing vesicles with the neuronal plasma membrane. Recent genetic studies have highlighted the critical role played by polyunsaturated fatty acids in neurotransmission, however, there is little information available about which fatty acids act on exocytosis and, more importantly, by what mechanism. We have used permeabilized chromaffin cells to screen various fatty acids of the n-3 and n-6 series for their acute effects on exocytosis. We have demonstrated that an n-6 series polyunsaturated fatty acid, arachidonic acid, potentiates secretion from intact neurosecretory cells regardless of the secretagogue used. We have shown that arachidonic acid dose dependently increases soluble NSF attachment protein receptor complex formation in chromaffin cells and bovine cortical brain extracts and that a non-hydrolysable analogue of arachidonic acid causes a similar increase in SNARE complex formation. This prompted us to examine the effect of arachidonic acid on SNARE protein interactions with Munc18a, a protein known to prevent Syntaxin1a engagement into the SNARE complex in vitro. In the presence of arachidonic acid, we show that Munc18a can interact with the neuronal SNARE complex in a dose-dependent manner. We further demonstrate that arachidonic acid directly interacts with Syntaxin1a.


[Use of the computer for management of a vaccination program].
The surveillance of vaccine acceptance was carried out in 3 French towns. The cost was minimal as the necessary equipment and personnel were available in the town-halls. This arrangement was utilized to monitor the increasing percentage of children vaccinated in the 3 towns. The increase was particularly significant for diphteria, tetanus and poliomyelitis.


Dynamic zinc pools in mouse choroid plexus.
We examined the presence of Zn-transporters (ZnT1, ZnT3, ZnT4, and ZnT6) proteins and zinc ions in rat choroid epithelium with immunohistochemistry and zinc selenide autometallography (ZnSe(AMG)). The four ZnT proteins were all expressed in the choroid epithelial cells. ZnT3 immunostaining was found in vesicle membranes in the apical part of the cells, associated to the microvillus membrane. Correspondingly, the ZnSe(AMG) technique revealed zinc ions in small vesicles, in microvilli, and multivesicular bodies in the epithelial cells. Traceable zinc ions were also found in lysosome-like organelles of fenestrated endothelial cells, but here no corresponding ZnT3 immunostaining was seen. The observations suggests that the choroid plexus is instrumental to regulation of the level of zinc ions in the cerebrospinal fluid.


Hypothalamic tanycytes: potential roles in the control of feeding and energy balance.
Tanycytes, glial-like cells that line the third ventricle, are emerging as components of the hypothalamic networks that control body weight and energy balance. They contact the cerebrospinal fluid (CSF) and send processes that come into close contact with neurons in the arcuate and ventromedial hypothalamic nuclei. Tanycytes are glucosensitive and are able to respond to transmitters associated with arousal and the drive to feed. At least some tanycytes are stem cells and, in the median eminence, may be stimulated by diet to generate new neurons. The quest is on to understand how tanycytes detect and respond to changes in energy balance and how they communicate with the rest of the nervous system to effect their functions.


Non-targeted 1H NMR fingerprinting and multivariate statistical analyses for the characterisation of the geographical origin of Italian sweet cherries.
In this study, non-targeted (1)H NMR fingerprinting was used in combination with multivariate statistical techniques for the classification of Italian sweet cherries based on their different geographical origins (Emilia Romagna and Puglia). As classification techniques, Soft Independent Modelling of Class Analogy (SIMCA), Partial Least Squares Discriminant Analysis (PLS-DA), and Linear Discriminant Analysis (LDA) were carried out and the results were compared. For LDA, before performing a refined selection of the number/combination of variables, two different strategies for a preliminary reduction of the variable number were tested. The best average recognition and CV prediction abilities (both 100.0%) were obtained for all the LDA models, although PLS-DA also showed remarkable performances (94.6%). All the statistical models were validated by observing the prediction abilities with respect to an external set of cherry samples. The best result (94.9%) was obtained with LDA by performing a best subset selection procedure on a set of 30 principal components previously selected by a stepwise decorrelation. The metabolites that mostly contributed to the classification performances of such LDA model, were found to be malate, glucose, fructose, glutamine and succinate.


Pharmacological ablation of astrocytes reduces Aβ degradation and synaptic connectivity in an ex vivo model of Alzheimer's disease.
Astrocytes provide a vital support to neurons in normal and pathological conditions. In Alzheimer's disease (AD) brains, reactive astrocytes have been found surrounding amyloid plaques, forming an astrocytic scar. However, their role and potential mechanisms whereby they affect neuroinflammation, amyloid pathology, and synaptic density in AD remain unclear. To explore the role of astrocytes on Aβ pathology and neuroinflammatory markers, we pharmacologically ablated them in organotypic brain culture slices (OBCSs) from 5XFAD mouse model of AD and wild-type (WT) littermates with the selective astrocytic toxin L-alpha-aminoadipate (L-AAA). To examine the effects on synaptic circuitry, we measured dendritic spine number and size in OBCSs from Thy-1-GFP transgenic mice incubated with synthetic Aβ42 or double transgenics Thy-1-GFP/5XFAD mice treated with LAAA or vehicle for 24 h. Treatment of OBCSs with L-AAA resulted in an increased expression of pro-inflammatory cytokine IL-6 in conditioned media of WTs and 5XFAD slices, associated with changes in microglia morphology but not in density. The profile of inflammatory markers following astrocytic loss was different in WT and transgenic cultures, showing reductions in inflammatory mediators produced in astrocytes only in WT sections. In addition, pharmacological ablation of astrocytes led to an increase in Aβ levels in homogenates of OBCS from 5XFAD mice compared with vehicle controls, with reduced enzymatic degradation of Aβ due to lower neprilysin and insulin-degrading enzyme (IDE) expression. Furthermore, OBSCs from wild-type mice treated with L-AAA and synthetic amyloid presented 56% higher levels of Aβ in culture media compared to sections treated with Aβ alone, concomitant with reduced expression of IDE in culture medium, suggesting that astrocytes contribute to Aβ clearance and degradation. Quantification of hippocampal dendritic spines revealed a reduction in their density following L-AAA treatment in all groups analyzed. In addition, pharmacological ablation of astrocytes resulted in a decrease in spine size in 5XFAD OBCSs but not in OBCSs from WT treated with synthetic Aβ compared to vehicle control. Astrocytes play a protective role in AD by aiding Aβ clearance and supporting synaptic plasticity.


Clonidine as an adjuvant for ultrasound guided supraclavicular brachial plexus block for upper extremity surgeries under tourniquet: A clinical study.
Clonidine has been used as an adjuvant to local anesthetic to extend the duration of block. The present study was aimed to compare the onset and duration of sensory and motor blockade of 0.75% ropivacaine alone or in combination with clonidine during ultrasound guided supraclavicular brachial plexus block for upper extremity surgeries under tourniquet. Sixty four adult American Society of Anesthesiologist grade 1 and 2 patients, scheduled for upper extremity surgeries were randomized to receive either 19.8 mL of 0.75% ropivacaine with 0.2 mL of normal saline (Group R) or 0.2 mL (30 μg) of clonidine (Group RC) in supraclavicular block. Onset and duration of sensory and motor blockade was compared. The hemodynamic variability, sedation, respiratory adequacy and any other adverse effects were also recorded. Ultrasound helped to visualize the nerves, needle and spread of local anesthetic at the brachial plexus block site. There was no statistically significant difference in the onset of sensory and motor blockade between the groups. Surgical anesthesia was achieved at the mean time of 20 min in all patients. Prolonged post-operative analgesia (mean duration 956 min) was observed in RC group as compared with R group (736 min). No complication of technique or adverse effect of ropivacaine and clonidine was reported. Clonidine as an adjuvant to ropivacaine for ultrasound guided supraclavicular brachial plexus enhanced duration of post-operative analgesia. There was no incidence of vessel puncture or pneumothorax.


OBSERVATIONS ON THREE SPECIES OF JELLYFISHES FROM CHESAPEAKE BAY WITH SPECIAL REFERENCE TO THEIR TOXINS. II. CYANEA CAPILLATA.
1. The human body exhibits little or no response to contact with the tentacles of specimens of Cyanea capillata from Chesapeake Bay. 2. Three types of nematocysts were identified: atrichous isorhizas, holotrichous isorhizas, and heterotrichous microbasic euryteles. 3. The tubes of nematocysts discharged into 2% agar take a sinuous course, often coiling on themselves. Most tubes penetrated less than 350µ. 4. Although faradic shock and FAA caused discharge of nematocysts from living tentacles, they failed to produce discharge of isolated nematocysts. 5. Homogenization of thoroughly washed frozen nematocysts causes rupture, releasing the toxic principle which is a single protein. 6. The LD 50 of the toxin was of the order of magnitude of µg/g of mouse body weight. 7. Mice were unaffected by intraperitoneal injections of washed capsular debris and tubes, or lipids, or of washed undischarged nematocysts in moderately heavy suspensions.


Total and split renal function in patients with renovascular hypertension: effects of angiotensin-converting enzyme inhibition.
Thirteen subjects with documented renovascular hypertension receiving chronic (greater than 1 month) therapy with an angiotensin-converting enzyme inhibitor (enalapril or captopril) underwent total and split renal function studies. Total glomerular filtration rate as assessed by inulin clearance was similar to that determined by radionuclide technique. Total effective renal plasma flow as assessed by p-aminohippurate clearance was lower than that determined by radionuclide technique. The glomerular filtration rate and effective renal plasma flow assessed by radionuclide technique of the stenotic kidney was comparatively lower than that of the non-stenotic kidney. No subject demonstrated complete loss of filtration or perfusion of the stenotic kidney. Five of six patients studied prospectively for 2 years have demonstrated stability of total renal function; the sixth patient, having a functional solitary stenotic kidney, has demonstrated stability of function following an initial abrupt decline in glomerular filtration rate and effective renal plasma flow. These results suggest that chronic angiotensin-converting enzyme inhibition therapy is not generally associated with near total absence of filtration of the stenotic kidney as has been suggested previously. Angiotensin-converting enzyme inhibitors may be safely and effectively utilized in the treatment of renovascular hypertension.


Profile of Instrumentation Laboratory's HemosIL® AcuStar HIT-Ab(PF4-H) assay for diagnosis of heparin-induced thrombocytopenia.
Immunoassays play an essential role in the diagnosis of heparin-induced thrombocytopenia (HIT). The objective of this article is to review HemosIL® AcuStar HIT-Ab(PF4-H) (Instrumentation Laboratory, Bedford, MA, USA), a new chemiluminescent immunoassay for HIT. Areas covered: The authors searched the published literature for evaluation studies of HemosIL® AcuStar HIT-Ab(PF4-H) and sought information from the manufacturer. In this paper, the authors discuss the analytical principle and technical aspects of the assay; describe its diagnostic performance in validation studies; report on its reproducibility, cost-effectiveness, and regulatory status; and discuss the implications of the assay on clinical practice and means of integrating it in diagnostic pathways. HemosIL® AcuStar HIT-Ab(PF4-H) is compared with other rapid assays and widely used enzyme-linked immunoassays for the diagnosis of HIT. Expert commentary: HemosIL® AcuStar HIT-Ab(PF4-H) is automatable, can be performed 24 h per day, offers a rapid turnaround time, and appears to have favorable diagnostic accuracy, particularly at thresholds above that listed in the label. These advantages could lead to improved patient outcomes through rapid provision of results at the point of care, enhancing the accuracy of initial diagnosis.


Blood-brain barrier permeability and electroconvulsive therapy - a systematic review.
The cause of cognitive side-effects after electroconvulsive therapy (ECT) is largely unknown. Alterations in the blood-brain barrier (BBB) has been considered in several recent ECT studies. We therefore found it worthwhile to perform a systematic review of the literature to examine if electrically induced seizures affect the permeability of the BBB. PubMed/MEDLINE and Embase were searched 16 November 2022. Studies with a direct measurement of BBB permeability in animals treated with modified electroconvulsive stimulation (ECS) and in humans treated with ECT were included. Synthesis of results were narrative due to the low number of studies and differences in study designs. Four animal and two human (31 participants) studies were included. In animals, two studies found increased BBB permeability to some smaller molecules after modified ECS, while the two other studies found marginally increased or unchanged permeability to albumin after treatment. In contrast, the human studies did not find increased BBB permeability to smaller molecules or albumin after ECT. Animal but not human studies support increased BBB permeability to some smaller molecules after electrically induced seizures. However, this conclusion is confined by the low number of studies and the lack of studies applying state-of-the art methods. More studies using modern approaches to measuring of BBB permeability are warranted.


Electro-fluorescence switching of bis-imidazolium ionic liquids.
Room-temperature ionic liquids (RTILs) containing bis-imidazolium salts were explored to control their optical properties of them in an organic device. The neat bis-imidazolium salts showed ionic conductivity of 3.5 x 10(-4) S/cm at room-temperature and the electrochemical window was exhibited within +/-2.5 V in a two electrode cell. The bis-imidazolium salts were transparent yellow and showed fluorescence upon excitation with light in the range of 360 to 500 nm. A two electrode organic ionic liquid cell was fabricated using a mixture of PEO200Blm-TFSI and electroactive molecules to control the emission property of the ionic liquid by electrochemical methods. The first example of the ionic liquid containing electro-fluorescence switch was explored with an electroactive benzyl viologen (BzV) blend of bis-imidazolium ionic liquid.


Yoruba culture and the resilience of HIV-positive adolescent girls in Nigeria.
Although there is a growing body of research exploring the influence of culture on the resilience of African youth, few studies have examined how culture constrains or enables resilience among HIV-positive adolescent girls from the perspective of the young women themselves. This paper reports on the findings from a qualitative study of five purposively selected girls living with HIV in Ibadan, Nigeria. By analysing data drawn mainly from interviews and observations, we explored how cultural influences promote or limit resilience in participants. Social-ecological resilience theory was used to document and interpret the findings. While some cultural values and perceptions enable resilience, others constrain participants' resilience trajectories. However, the girls were able to navigate through these constraints using their cultural identities and coping strategies, such as future dreams, emotional and physical resources linked to spirituality and networks of friends and families. Findings have implications for policymakers, researchers and programmers in strengthening the health and resilience of young people in the face of HIV.


Selective Predation by Owls on Infected Bank Voles (Myodes glareolus) as a Possible Sentinel of Tularemia Outbreaks.
Tularemia is a widely spread zoonotic disease in the northern hemisphere, caused by the bacterium Francisella tularensis. In humans, tularemia is an acute febrile illness with incidence peaks in late summer to early autumn of outbreak years, but there is no early warning system in place that can reduce the impact of disease by providing timely risk information. In this study, we revisit previously unpublished data on F. tularensis in water, sediment, soil, and small mammals from 1984 in northern Sweden. In addition, we used human case data from the national surveillance system for tularemia in the same year. In the environmental and small mammal material, bank vole (Myodes glareolus) samples from urine and bladder were the only samples that tested positive for F. tularensis. The prevalence of F. tularensis among trapped bank voles was 13.5%, although all six bank voles that were retrieved from owl nest boxes in early May tested positive. Forty-two human tularemia cases were reported from August to December in 1984. Based on these results, we encourage investigating the potential role of tularemia-infected bank voles retrieved from owl nest boxes in spring as an early warning for outbreaks of tularemia among humans in summer and autumn of the same year.


[Antibiotic treatment as the standard for appendicitis].
Acute appendicitis has traditionally been treated surgically. Conservative approach with antibiotic treatment has been suggested as an alternative to the surgical procedure. The available studies on surgery versus antibiotics may indicate the safe use of conservative treatment for uncomplicated appendicitis, but the available studies are typically of low quality. Thus, further studies are needed to clarify the role of conservative treatment, and in the meantime appendectomy remains the standard treatment for acute appendicitis.


Receptive field properties of the macaque second somatosensory cortex: nonlinear mechanisms underlying the representation of orientation within a finger pad.
We investigate the position invariant receptive field properties of neurons in the macaque second somatosensory (SII) cortical region. Previously we reported that many SII region neurons show orientation tuning in the center of multiple finger pads of the hand and further that the tuning is similar on different pads, which can be interpreted as position invariance. Here we study the receptive field properties of a single finger pad for a subset (n = 61) of those 928 neurons, using a motorized oriented bar that we positioned at multiple locations across the pad. We calculate both vector fields and linear receptive fields of the finger pad to characterize the receptive field properties that give rise to the tuning, and we perform an additional regression analysis to quantify linearity, invariance, or both in individual neurons. We show that orientation tuning of SII region neurons is based on a variety of mechanisms. For some neurons, the tuning is explained by simple excitatory regions, simple inhibitory regions, or some combination of these structures. However, a large fraction of the neurons (n = 20 of 61, 33%) show position invariance that is not explained well by their linear receptive fields. Finding invariance within a finger pad, coupled with the previous result of similar tuning on different pads, indicates that some SII region neurons may exhibit similar tuning throughout large regions of the hand. We hypothesize that invariant neurons play an important role in tactile form recognition.


Meeting the governance challenges of next-generation biorepository research.
Advances in clinical translational research have led to an explosion of interest in infrastructure development and data sharing facilitated by biorepositories of specimens and linked health information. These efforts are qualitatively different from the single-center sample collections that preceded them and pose substantial new ethics and regulatory challenges for investigators and institutions. New research governance approaches, which can address current and anticipated challenges, promote high-quality research, and provide a robust basis for ongoing research participation, are urgently required.


Newly Designed Silica-Containing Redox Nanoparticles for Oral Delivery of Novel TOP2 Catalytic Inhibitor for Treating Colon Cancer.
Although oral drug delivery is the most common route of drug administration, the conventional polymeric nanocarriers exhibit a low drug loading capacity and low stability in the gastrointestinal (GI) environments. In this study, a newly designed silica-containing redox nanoparticle (siRNP) with reactive oxygen species (ROS) scavenging capacity is developed as an ideal oral nanocarrier for a novel hydrophobic anticancer compound BNS-22 to treat colitis-associated colon cancer in vivo. Crosslinking of silica moieties significantly enhances the stability under acidic conditions and improves BNS-22 loading capacity of siRNP compared to the conventional redox nanoparticle. After oral administration to mice, BNS-22-loaded siRNP (BNS-22@siRNP) remarkably improves bioavailability and colonic tumor distribution of BNS-22. As the result, BNS-22@siRNP significantly inhibits the tumor progression in colitis-associated colon cancer mice compared to other control treatments. It is noteworthy that no systemic absorption of siRNP carrier is observed after oral administration. Interestingly, orally administered BNS-22@siRNP significantly suppresses the adverse effects of BNS-22 owing to its ROS scavenging capacity, and no other noticeable toxicities are observed in mice treated with BNS-22@siRNP although siRNP is localized in the GI tract. Our results indicate that siRNP is a promising oral drug nanocarrier for cancer therapy.


Sinus pericranii: long-term outcome in a 10-year-old boy with a review of literature.
Sinus pericranii is a rare cranial venous anomaly in which there is an abnormal communication between intracranial dural sinuses and extracranial venous structures, usually via an emissary transosseous vein. We report the case of a 10-year-old boy who was successfully managed conservatively. The clinical findings, investigations and management are discussed. We focus on the long-term management of these patients, with a review of the relevant literature.


Social skills assessment of children with autism in free-play situations.
Poor social functioning and limited play are characteristic of children with autism. Increasingly, education for children with autism is provided within mainstream settings, but given their particular difficulties, the adequate provision of educational services in such settings is challenging. This study presents observational data of the play behaviour and social interaction patterns of 10 children with autism in mainstream kindergartens and primary school playgrounds. The target children differed significantly in terms of their play and social interactions from typically developing children in the same settings. The adequacy of the provision of services for children with autism in mainstream provision is discussed.


Antibody to streptococcal cell wall peptidoglycan-polysaccharide polymers in seropositive and seronegative rheumatic disease.
An ELISA has been developed for serum antibodies to streptococcal cell wall peptidoglycan-polysaccharide polymers (PG-GSP). A significantly increased prevalence of serum anti-PG-GSP antibody was found in juvenile chronic arthritis and both seropositive and seronegative rheumatoid arthritis (RA), compared with ankylosing spondylitis, systemic lupus erythematosus, myeloma and healthy controls. Anti-PG-GSP antibody was always of the IgG class and there was no correlation of anti-PG-GSP levels with C reactive protein, rheumatoid factor (RF) or anti-streptolysin O titres. There was no direct cross-reaction of RF with PG-GSP, nor did the presence of IgM-RF significantly interfere with the assay. Examination of paired serum and synovial fluid samples offered no evidence for local production of anti-PG-GSP antibody in synovial tissue. These data are compatible with an increased systemic immunization by bacterial fragments in RA.


Pyrrolo[2,3-d]Pyrimidines as Kinase Inhibitors.
The pyrrolo[2,3-d]pyrimidine nucleus is a deaza-isostere of adenine, the nitrogenous base of ATP, and is present in many ATP-competitive inhibitors of different kinases. In the last few years the number of articles and patents that have appeared involving this type of inhibitors has dramatically increased and some compounds have been approved for the treatment of inflammatory or myeloproliferative diseases. Other derivatives are currently being evaluated in clinical trials. This review deals with pyrrolo[2,3- d]pyrimidine derivatives active as kinase inhibitors that have been reported in the literature from 2011 to 2016, with a particular interest on the recently patented compounds. The molecules are classified depending on the inhibited kinase, focusing on their chemical structures.


A Nutrient-Sensing Transition at Birth Triggers Glucose-Responsive Insulin Secretion.
A drastic transition at birth, from constant maternal nutrient supply in utero to intermittent postnatal feeding, requires changes in the metabolic system of the neonate. Despite their central role in metabolic homeostasis, little is known about how pancreatic β cells adjust to the new nutritional challenge. Here, we find that after birth β cell function shifts from amino acid- to glucose-stimulated insulin secretion in correlation with the change in the nutritional environment. This adaptation is mediated by a transition in nutrient sensitivity of the mTORC1 pathway, which leads to intermittent mTORC1 activity. Disrupting nutrient sensitivity of mTORC1 in mature β cells reverts insulin secretion to a functionally immature state. Finally, manipulating nutrient sensitivity of mTORC1 in stem cell-derived β cells in vitro strongly enhances their glucose-responsive insulin secretion. These results reveal a mechanism by which nutrients regulate β cell function, thereby enabling a metabolic adaptation for the newborn.


[Esthetic evaluation of conventional and socket shield technique immediate implant restoration].
Objective: To evaluate the pink and white esthetics of conventional and socket shield technique (SST) immediate implant restoration. Methods: Thirty volunteers were recruited according to preset criteria, and were assigned to 3 groups. Natural teeth group (ten undergraduates or postgraduates from Peking University School and Hospital of Stomatology in January 2020, n=10): all volunteers' maxillary anterior teeth were natural teeth with healthy gingiva, and none of the teeth were restored by crowns or composite. Conventional group and SST group (patients had a maxillary central incisor immediate implant placed in Department of Prosthodontics, Peking University School and Hospital of Stomatology during October 2016 to January 2021, n=10 for each group): the volunteer had an unrestored natural maxillary central incisor, and the contralateral maxillary central incisor was restored by conventional or SST immediate implant placement, temporization and all ceramic final restoration, photos were taken 12 months post-surgery. Three groups of evaluators namely layperson (staff from Second Clinical Division of Peking University School and Hospital of Stomatology and 2 family members, n=10), dental students (class 2015 undergraduates from Peking University School and Hospital of Stomatology, n=10) and prosthodontists (from Department of Prosthodontics, Peking University School and Hospital of Stomatology, n=10) were invited to assess the esthetics using pink esthetic score (PES) and white esthetic score (WES). The results were statistically analyzed. Results: PES and WES of natural teeth group [9(8, 10) and 8(7, 10)] were significantly higher than conventional group [7(6,8) and 7(6,9)] (H=287.08, 132.79,P<0.01) and SST group [7(6, 9) and 8(7, 9)] (H=216.01, 101.21, P<0.01). SST group yielded higher PES than the conventional group (H=-71.06, P<0.01), yet had similar WES (H=-31.57, P>0.05). Dental students had significant lower PES and WES than prosthodontists (H=-120.90, -218.86, P<0.01) and layperson (H=-109.55, 134.97, P<0.01). Prosthodontists and layperson got similar PES (H=-11.36, P>0.05), however yielded different WES (H=-83.89, P<0.01). Conclusions: SST immediate implant placement obtained better pink esthetics than conventional protocol 12-month after implant surgery, profession may have significant impact on evaluators during pink and white esthetic evaluation.


Health equity, aboriginal peoples and occupational therapy.
In Canada, Aboriginal peoples are affected by colonial relations of power that result in poor health outcomes. Despite occupational therapists' efforts to work in a safe and competent manner with people experiencing challenges to participating in daily life, Western healthcare models reflect values that often undermine Aboriginal peoples' health and well-being. Meaningful, effective, and culturally appropriate healthcare practices cannot be fully implemented by occupational therapists without an ongoing and critical examination of occupational therapy's foundational belief systems. Only a critical examination of these foundational belief systems will enable occupational therapists to take action towards addressing these inequities, which is an important step in moving towards culturally safe care. Canadian health professions, including occupational therapy, have the potential to create positive change at a systems level through the critical exploration of underlying professional assumptions. To advance dialogue about Aboriginal peoples' health, occupational therapists must engage in exploration of their profession's underlying theoretical concepts or risk participating in the perpetuation of health inequities for already at-risk populations.


Lactoferrin- and Dendrimer-Bearing Gold Nanocages for Stimulus-Free DNA Delivery to Prostate Cancer Cells.
The use of gene therapy to treat prostate cancer is hampered by the lack of effective nanocarriers that can selectively deliver therapeutic genes to cancer cells. To overcome this, we hypothesize that conjugating lactoferrin, a tumor-targeting ligand, and the diaminobutyric polypropylenimine dendrimer into gold nanocages, followed by complexation with a plasmid DNA, would enhance gene expression and anti-proliferation activity in prostate cancer cells without the use of external stimuli. Novel gold nanocages bearing lactoferrin and conjugated to diaminobutyric polypropylenimine dendrimer (AuNCs-DAB-Lf) were synthesized and characterized. Following complexation with a plasmid DNA, their gene expression, cellular uptake and anti-proliferative efficacies were evaluated on PC-3 prostate cancer cells. AuNCs-DAB-Lf was able to complex DNA at conjugate: DNA weight ratios 5:1 onwards. Gene expression was at its highest after treatment with AuNCs-DAB-Lf at a weight ratio of 10:1, as a result of a significant increase in DNA uptake mediated by the conjugate at that ratio in PC-3 cells. Consequently, the anti-proliferative activity of AuNCs-DAB-Lf-DNA encoding TNFα was significantly improved by up to 9-fold compared with DAB dendriplex encoding TNFα. Lactoferrin-bearing dendrimer-conjugated gold nanocages are highly promising gene delivery systems for the treatment of prostate cancer.


Predictors of costs in dementia in a longitudinal perspective.
To analyse predictors of costs in dementia from a societal perspective in a longitudinal setting. Healthcare resource use and costs were assessed retrospectively using a questionnaire in four waves at 6-month intervals in a sample of dementia patients (N = 175). Sociodemographic data, dementia severity and comorbidity at baseline, cognitive impairment and impairment in basic and instrumental activities of daily living were also recorded. Linear mixed regression models with random intercepts for individuals were used to analyse predictors of total and sector-specific costs. Impairment in activities of daily living significantly predicted total costs in dementia patients, with associations between basic activities of daily living and formal care costs on the one and instrumental activities of daily living and informal care costs on the other hand. Nursing home residence was associated with lower total costs than residence in the community. There was no effect of cognition on total or sector-specific costs. Cognitive deficits in dementia are associated with costs only via their effect on the patients' capacity for activities of daily living. Transition into a nursing home may reduce total costs from a societal perspective, owing to the fact that a high amount of informal care required by severely demented patients prior to transition into a nursing home may cause higher costs than inpatient nursing care.


Wolf (Canis lupus) Generation Time and Proportion of Current Breeding Females by Age.
Information is sparse about aspects of female wolf (Canis lupus) breeding in the wild, including age of first reproduction, mean age of primiparity, generation time, and proportion of each age that breeds in any given year. We studied these subjects in 86 wolves (113 captures) in the Superior National Forest (SNF), Minnesota (MN), during 1972-2013 where wolves were legally protected for most of the period, and in 159 harvested wolves from throughout MN wolf range during 2012-2014. Breeding status of SNF wolves were assessed via nipple measurements, and wolves from throughout MN wolf range, by placental scars. In the SNF, proportions of currently breeding females (those breeding in the year sampled) ranged from 19% at age 2 to 80% at age 5, and from throughout wolf range, from 33% at age 2 to 100% at age 7. Excluding pups and yearlings, only 33% to 36% of SNF females and 58% of females from throughout MN wolf range bred in any given year. Generation time for SNF wolves was 4.3 years and for MN wolf range, 4.7 years. These findings will be useful in modeling wolf population dynamics and in wolf genetic and dog-domestication studies.


Optical Detection of Distal Lung Enzyme Activity in Human Inflammatory Lung Disease.
Objective and Impact Statement. There is a need to develop platforms delineating inflammatory biology of the distal human lung. We describe a platform technology approach to detect in situ enzyme activity and observe drug inhibition in the distal human lung using a combination of matrix metalloproteinase (MMP) optical reporters, fibered confocal fluorescence microscopy (FCFM), and a bespoke delivery device. Introduction. The development of new therapeutic agents is hindered by the lack of in vivo in situ experimental methodologies that can rapidly evaluate the biological activity or drug-target engagement in patients. Methods. We optimised a novel highly quenched optical molecular reporter of enzyme activity (FIB One) and developed a translational pathway for in-human assessment. Results. We demonstrate the specificity for matrix metalloproteases (MMPs) 2, 9, and 13 and probe dequenching within physiological levels of MMPs and feasibility of imaging within whole lung models in preclinical settings. Subsequently, in a first-in-human exploratory experimental medicine study of patients with fibroproliferative lung disease, we demonstrate, through FCFM, the MMP activity in the alveolar space measured through FIB One fluorescence increase (with pharmacological inhibition). Conclusion. This translational in situ approach enables a new methodology to demonstrate active drug target effects of the distal lung and consequently may inform therapeutic drug development pathways.


Transition of care in patients with anorectal malformations: Consensus by the ARM-net consortium.
To develop the first consensus to standardize the management of patients with Anorectal Malformations (ARMs) transitioning from childhood to adulthood. A dedicated task force of experts performed an extensive literature review and multiple meetings to define the most important aspects of transition of care. The findings were discussed with all ARM-net consortium members and a set of practical recommendations agreed upon at the annual meeting in 2016. We defined seven domains that are essential to provide an effective and practical transition process. Within each domain we have developed a set of key recommendations that are important to be considered for ARM patients entering the age of transition. It is crucial that transition begins at an early age with regular and well-structured follow-up. Cooperation with a selected multidisciplinary team of pediatric and adult practitioners is required to prepare patients and families for effective transition to adult care and to reduce long term morbidity. Review/Consensus paper. III.


Quantifying the Susceptibility Variation of Normal-Appearing White Matter in Multiple Sclerosis by Quantitative Susceptibility Mapping.
The purpose of this study is to evaluate the magnetic susceptibility of normal-appearing white matter (NAWM) in patients with multiple sclerosis (MS) using quantitative susceptibility mapping. Ninety-four patients with relapse-remitting MS (RRMS) (37 with gadolinium-enhancing lesions and 57 with only gadolinium-nonenhancing lesions) and 55 healthy control subjects were included in this retrospective study. The susceptibility values of NAWM relative to CSF in patients with MS were compared with those of white matter (WM) in healthy control subjects and were correlated with the patient status of gadolinium-enhancing lesions, disease duration, and expanded disability status scale scores. All 37 patients with RRMS and gadolinium-enhancing lesions also had gadolinium-nonenhancing lesions. Susceptibility values of NAWM in patients with MS and only gadolinium-nonenhancing lesions (-18.29 ± 8.03 parts per billion [ppb]) were higher than those for WM in healthy control subjects (-25.81 ± 6.02 ppb; p < 0.001) and NAWM in patients with gadolinium-enhancing lesions (-25.64 ± 6.55 ppb; p < 0.001). Susceptibility values of NAWM in patients with MS with gadolinium-enhancing lesions were similar to those for WM in healthy control subjects (p = 0.91). This trend was dependent on neither NAWM region nor disease duration when the data were controlled for age. NAWM susceptibility was not correlated with either disease duration or expanded disability status scale (p > 0.05). In patients with RRMS and gadolinium-nonenhancing lesions, the susceptibility values of NAWM decrease when gadolinium-enhancing lesions appear, approaching values similar to those of WM in healthy control subjects, suggesting that NAWM may contribute to the iron accumulation observed in early active MS lesions.


Long-term, open-label, safety study of once-daily ropinirole extended/prolonged release in early and advanced Parkinson's disease.
Long-term safety of once-daily ropinirole extended/prolonged release (ropinirole XL/PR) was evaluated in subjects with early and advanced Parkinson's disease (PD) in this study, 101468/248. Subjects (n = 419) who completed one of three prior studies evaluating ropinirole XL/PR for the treatment of PD were enrolled in this open-label, multicenter, extension study, and were to be followed for up to 73 months. Ropinirole XL/PR was titrated/continued, and adjusted as appropriate during the maintenance phase (maximum 24 mg/d). Levodopa (L-dopa) and other nondopamine agonist PD medications were permitted. Safety outcomes that were investigated included frequency of adverse events (AEs). Subjects' preference regarding once daily versus three times daily study medication regimens was also investigated in a subset of the study population. The median duration of ropinirole XL/PR exposure was 1275 d. Most subjects (87%) reported at least one AE, with the most common (≥ 10%) AEs being, back pain (14%), hallucinations (13%), somnolence (11%) and peripheral edema (11%). Twenty-five percent of subjects discontinued the study prematurely due to an AE during the treatment period. Long-term treatment with ropinirole XL/PR was not associated with any new or unexpected safety concerns in patients with early and advanced PD, and a majority of subjects preferred the once-daily dosing regimen.


Polymerase chain reaction of pleural biopsy specimens for rapid diagnosis of tuberculous pleuritis.
Government-administered regional teaching hospital. To improve timeliness and sensitivity of laboratory diagnosis of tuberculous pleuritis. We applied polymerase chain reaction (PCR) to detect DNA (IS6110) specific for Mycobacterium tuberculosis complex in pleural biopsy specimens. Of 28 patients with pleural disease, 11 were diagnosed by microbiology (smear/culture of sputum or pleural fluid) with tuberculous pleuritis, eight were diagnosed with tuberculous pleuritis by histology (of pleural biopsies) and/or clinical presentation, and nine were diagnosed with carcinomatous pleuritis. Seventeen of the patients' pleural biopsies were PCR positive. Based on microbiological results, the sensitivity of the PCR assay was 100% (11/11). On the other hand, based on the results of the histological and clinical data, sensitivity and specificity of the PCR results were 89% (17/19) and 100% (9/9), respectively. PCR of pleural biopsy specimens can be a useful method when employed in combination with microbiological and histological examinations of pleural biopsy for rapid diagnosis of tuberculous pleuritis.


Laparoscopic treatment for appendicitis during pregnancy: Retrospective cohort study.
Acute appendicitis is the most frequent non-obstetric surgical emergency during pregnancy. The benefits of laparoscopy during pregnancy are well known, but complications can occur, and these can affect both the mother and/or the foetus.We present results of laparoscopic surgical treatment of acute appendicitis in pregnant women, analysing the occurrence of adverse postoperative, obstetric and foetal outcomes and reviewing literature. Retrospective observational study on pregnant women with a preoperative diagnosis of acute appendicitis. n = 63, mean age 28.4 years, average gestational age of 17.7 weeks (3-30 weeks). 6.4 % exploratory laparoscopies, 92 % laparoscopic appendectomies and one right colectomy were performed. Conversion rate was 3.2 %. When symptoms begun within 48 hours prior to surgery, a perforated appendicitis was found in 11 %; whereas when the time from symptom onset to surgery was greater than or equal to 48 hours, it was evident in 31 % of the cases (p 0.008). The only independent variable associated with the presence of postoperative complications was symptom duration prior to surgery greater than or equal to 48 hours (OR 4.8; 95 % CI 1.1-16.2; p 0.04). Seven minor and 2 mayor postoperative complications were observed. Patients with complications spent, on average, twice as many days hospitalized (p < 0.001); and had 8 times more risk of preterm delivery (p 0.03). Obstetric complications were more frequent in pregnant women operated during the first trimester. Foetal mortality was 1.6 %. Surgical morbidity of acute appendicitis in pregnant women is linked to the delay in the diagnosis and treatment of the inflammatory condition. Laparoscopic appendectomy during pregnancy is not exempt from postoperative, obstetric and foetal complications. It is necessary to standardize the definitions of "complication" in order to collate reliably the outcomes presented in the literature.


On the Molecular Basis of Division of Labor in Solenopsis invicta (Hymenoptera: Formicidae) Workers: RNA-seq Analysis.
The fire ant Solenopsis invicta Buren is an important invasive pest. Among S. invicta workers behavioral changes depend on age where younger ants are nurses and older ants foragers. To identify potential genes associated with this division of labor, we compared gene expression between foragers and nurses by high-throughput sequencing. In total, we identified 1,618 genes significantly differently expressed between nurses and foragers, of which 542 were upregulated in foragers and 1,076 were upregulated in nurses. Several pathways related to metabolism were significantly enriched, such as lipid storage and fatty acid biosynthesis, which might contribute to the division of labor in S. invicta. Several genes involved in DNA methylation, transcription, and olfactory responses as well as resistance to stress were differentially expressed between nurses and foragers workers. Finally, a comparison between previously published microarray data and our RNA-seq data in S. invicta shows 116 genes overlap, and the GO term myofibril assembly (GO: 0030239) were simultaneously significantly enriched. These results advance knowledge of potentially important genes and molecular pathways associated with worker division of labor in S. invicta. We hope our dataset will provide . candidate target genes to disrupt organization in S. invicta as a control strategy against this invasive pest.


Recombinant expression of indolicidin concatamers in Escherichia coli.
Antimicrobial peptides are part of the innate immune system of vertebrates and invertebrates. They are active against gram-negative and gram-positive bacteria, fungi, and protozoa. Currently, most antimicrobial peptides are extracted from host organisms or produced by solid-phase peptide synthesis. Recombinant protein expression in Escherichia coli is a tool for greater production yields at a decreased cost and reduces the use of hazardous materials. We have constructed a concatamer of indolicidin and successfully expressed a fusion product with thioredoxin in E. coli BL21DE3. Codons for methionine residues flanking individual indolicidin genes were incorporated for cyanogen bromide cleavage of the fusion protein and liberation of active monomeric indolicidin. Peptide yields of 150 microg/l monomeric indolicidin were achieved in this first report of recombinant production of indolicidin with demonstrated antimicrobial activity.


Does airborne ultrasound lead to activation of the auditory cortex?
As airborne ultrasound can be found in many technical applications and everyday situations, the question as to whether sounds at these frequencies can be heard by human beings or whether they present a risk to their hearing system is of great practical relevance. To objectively study these issues, the monaural hearing threshold in the frequency range from 14 to 24 kHz was determined for 26 test subjects between 19 and 33 years of age using pure tone audiometry. The hearing threshold values increased strongly with increasing frequency up to around 21 kHz, followed by a range with a smaller slope toward 24 kHz. The number of subjects who could respond positively to the threshold measurements decreased dramatically above 21 kHz. Brain activation was then measured by means of magnetoencephalography (MEG) and functional magnetic resonance imaging (fMRI) and with acoustic stimuli at the same frequencies, with sound pressure levels (SPLs) above and below the individual threshold. No auditory cortex activation was found for levels below the threshold. Although test subjects reported audible sounds above the threshold, no brain activity was identified in the above-threshold case under current experimental conditions except at the highest sensation level, which was presented at the lowest test frequency.


Slide tracheoplasty in an infant with congenital tracheal stenosis and oesophageal atresia with tracheoesophageal fistula.
Slide tracheoplasty has become the treatment of choice when dealing with congenital tracheal stenosis (CTS). Versatility and effectiveness are the specific advantages of this technique when compared with other surgical procedures for tracheal reconstruction. We report a new use of slide tracheoplasty in a patient with a short CTS and a tracheal pouch after oesophageal atresia with tracheoesophageal fistula repair.


Review of the measuring precision of the new Meibometer MB550 through repeated measurements in dogs.
A meibometer is a device to measure the delivery rate of lipids on the eyelid margin. The aim of this study is to determine the measuring precision of the new Meibometer MB550 (Courage-Khazaka electronic GmbH, 50829, Cologne, Germany), linked to a computer, by means of repeated measurements in dogs by different examiners. Two investigators measured the lipid rate on the eyelid margin in 10 healthy dogs for 10 days. One examiner measured the right eye (OD) and the other measured the left eye (OS) for 5 days. After 5 days, the eyes to be measured were switched between the examiners. The new device was able to record all measurement values as charts and curves in comparison to the previous Meibometer, which displayed only one value. Mean meibomium lipid level +/- SD in the OD and OS of 10 dogs in 10 days was 211 +/- 48 MU and 205 +/- 41 MU (meibometer units), respectively. There was no significant difference between OD and OS, but a broad distribution of the values could be found. Gender did not influence the measurements. A significant difference was determined between the measurements in the first 5 days to those in the following 5 days. There was, therefore, also a significant difference in the measurements of the 2 examiners. Meibometry is a minimally invasive lipid measuring method on the eyelid margin. Repeated measurement results obtained by two examiners, with the new device Meibometer MB550 linked to a computer, showed a wide range of values. The measuring precision of the new Meibometer MB550 is therefore questionable.


A modified version of diagonal systematic sampling in the presence of linear trend.
Systematic sampling is one of the simplest and popular methods for selecting a random sample from a finite population. The diagonal systematic sampling scheme is a type of systematic sampling design which has gained the attention of researchers during the last two decades. In this paper, a modification to the conventional diagonal systematic sampling design is proposed for use in situations where population units follow a linear trend. It is found that the proposed strategy reduces the variance of the diagonal systematic sampling thus resulting in an efficient sampling design. The mathematical conditions under which the suggested modified diagonal systematic sampling design is more precise than some of the available sampling designs are derived. With the help of a numerical illustration using milk yield data, it is shown that the proposed sampling scheme is more efficient than some of the available sampling schemes.


An Exonuclease I-Based Quencher-Free Fluorescent Method Using DNA Hairpin Probes for Rapid Detection of MicroRNA.
MicroRNAs (miRNAs) act as biomarkers for the diagnosis of a variety of cancers. Since the currently used methods for miRNA detection have limitations, simple, sensitive, and cost-effective methods for the detection of miRNA are required. This work demonstrates a facile, quencher-free, fluorescence-based analytical method for cost-effective and sensitive detection of miRNA using a super 2-aminopurine (2-AP)-labeled hairpin probe (HP) and exonuclease I activity. Specifically, the fluorescence of 2-AP is strongly quenched when it is incorporated within DNA. In the presence of a target miRNA, HP attains an open conformation by hybridizing with the target miRNA to form a double-stranded structure with a protruding 3'-terminus. Next, the digestion of the protruding 3'-terminus is triggered by exonuclease I, during which 2-AP is released free in solution from the DNA, thereby increasing fluorescence. This method is highly sensitive, with a detection limit of 0.5 nM-10 times lower than a previously reported quencher-free fluorescence method. Furthermore, this method has potential applications in clinical diagnosis and biomedical research.


Rabies in African wild dogs (Lycaon pictus) in the Serengeti region, Tanzania.
Rabies was confirmed as the cause of death of one African wild dog (Lycaon pictus) in the Serengeti region, Tanzania. One adult African wild dog in the same pack showed central nervous signs consistent with rabies infection. Inactivated rabies vaccine was administered intramuscularly to African wild dogs in two packs, by dart or by hand following anesthesia. These individuals comprised all known adults in the Serengeti National Park. In a limited study of seroprevalence of rabies antibody carried out at the time of vaccination, 3 of 12 African wild dogs sampled in the Serengeti had rabies serum neutralizing antibody titers before vaccination. Paired serum samples from two individuals sampled after vaccination showed increased antibody titers.


[Optimal parameters of an adaptive temporal selector of informative elements of the electrocardiosignals].
The adaptive temporal selector increases 2-3 times the noise immunity of detectors of ECG informative elements, especially of R = R-wave detectors, that is of particular importance when patient moves. The selector produces the protective time interval the duration of which is conditioned by the duration of the preceeding R = R-interval, so the probability of false detections caused by noise is reduced. The optimum protective time interval should constitute 84 per cent of the preceeding R = R-interval, the error should not exceed +/- 5%.


The Unfinished Business of Respect for Autonomy: Persons, Relationships, and Nonhuman Animals.
This essay explores three issues in respect for autonomy that pose unfinished business for the concept. By this, I mean that the dialogue over them is ongoing and essentially unresolved. These are: (1) whether we ought to respect persons or their autonomous choices; (2) the role of relational autonomy; and (3) whether nonhuman animals can be autonomous. In attending to this particular set of unfinished business, I highlight some critical moral work left aside by the concept of respect for autonomy as understood in Beauchamp and Childress' Principles of Biomedical Ethics. Specifically, while significant pragmatic traction is gained by the authors' focus on autonomous choice, carving such a focus out from the broader questions of moral respect and the autonomy of the person leaves aside a number of questions that we might have thought a view about respect for autonomy in biomedicine ought to answer. These include: How should physicians respond when autonomous patients make decisions that appear nonautonomous? What is the impact of the view that autonomy is "relational" for cross-cultural differences in how autonomy is respected? If chimpanzees (and by extension young children) can be autonomous, what does that mean for how they should be treated?


Maturation-related differences in the structure and composition of proteoglycans synthesized by chondrocytes from bovine articular cartilage.
Calf (2-3-month-old) and steer (approximately 18-month-old) bovine articular chondrocytes were isolated and cultured as high density monolayers. The proteoglycans synthesized on day 5 during a 15-h period of labeling with [35S]sulfate or [3H]glucosamine were isolated and characterized. The majority (greater than 70%) of the newly synthesized proteoglycans were found in the medium. When viewed in the electron microscope, medium-derived proteoglycans of high buoyant density were longer in calf than in steer. The medium and extracts of the cell layer were pooled and the radiolabeled proteoglycans were fractionated by isopycnic density gradient centrifugation performed under dissociative conditions. The low buoyant density fraction contained, in both calf and steer, small-sized nonaggregating proteoglycans containing chondroitin sulfate. The high buoyant density fraction contained greater than 90% of the newly synthesized proteoglycans. The majority were able to interact with hyaluronic acid to form aggregates. Calf high buoyant density fraction proteoglycans were larger, had longer chondroitin sulfate chains and lower ratios of keratan sulfate chains/chondroitin sulfate chains than steer high buoyant density fraction proteoglycans. These maturation-related differences are typical of those present in the proteoglycans of the calf and steer cartilage matrix from which the chondrocytes were isolated. Experiments with beta-D-xylosides showed that steer cultures had the capacity to synthesize twice as many chondroitin sulfate chains/cell as calf cultures. At each xyloside concentration used, chondroitin sulfate chains were longer in calf than steer. At both ages, chain size decreased with increase in rate of synthesis; the relationship between chain size and rate of synthesis was, however, quite different at the two ages. The results of these studies suggest that articular chondrocytes have an inherent program that determines the quality of proteoglycans synthesized at different ages.


Spontaneous regression of bovine cutaneous leukosis.
Biopsies of skin from a 2-year-old heifer with spontaneously regressing dermal leukosis were examined. The heifer was not infected with bovine leukemia virus and was negative for tumor-associated antigens of enzootic bovine lymphosarcoma. By hematological standards for bovine leukemia, the heifer was positive at about 60 days post-occurrence of the disease (POD). At 53 days POD, lymphoblastic neoplastic cells in the dermis reacted with anti-T lymphocyte monoclonal antibody by the avidin biotin peroxidase complex method. The cells had intracytoplasmic clustered dense bodies under electron microscopy. From 53 to 83 days POD, figures of the transepithelial elimination (TE) against neoplastic cells and perivascular infiltration of small lymphocytes were in the dermis. Small lymphocytes reacted with anti-T lymphocyte monoclonal antibody. At necropsy there were no neoplastic lesions; there were flat lymph node-like tissues in the subcutis. Many germinal centers were seen in the lymphatic organs. Blood lymphocytes at 46 days POD were stimulated by phytohemagglutinin-P and concanavalin A. Sera, taken until 75 days POD and at necropsy, showed an inhibitory effect on mitogen-induced blastogenesis of normal bovine lymphocytes. These results suggested the existence of a spontaneous regressive mechanism against neoplastic lesions by TE and tumor immunity.


Comparative antioxidant and cytotoxic effect of procyanidin fractions from grape and pine.
There is a great interest in characterizing the biological properties of natural compounds obtained from plants, especially polyphenols. We studied the structure-activity-cytotoxicity relationships of polyphenolic fractions obtained from grape pomace and pine bark. These fractions contained similar polymerised flavonoids but different percentages of pyrogallol groups that confer on them different biological properties. The human keratinocyte cell line HaCaT and the mouse fibroblast cell line 3T3 were used to study the cytotoxicity of the different fractions after 24, 48, and 72 h of exposure. Antioxidant activity of the fractions was evaluated by measuring the inhibition of hemolysis mediated by AAPH. Our results demonstrate that the polyphenolic fractions studied show high antioxidant capacity in a concentration range that is not harmful to normal human cells. Pine fractions presented slightly lower antioxidant activity than grape fractions but are less cytotoxic. This data provides useful information to help design safe antioxidant products that act without altering critical cell functions.


Co-expression of CD24 and Hsp70 as a prognostic biomarker for lung cancer.
Lung cancer is one of the most common malignant neoplasms worldwide. CD24 is a marker of tumor stem cells that plays an important role in tumorigenesis. Hsp70 is an important molecular chaperone. However, the co-expression and interaction of CD24 and Hsp70, as well as the significance for the prognosis of lung cancer are still unclear. The expression levels of CD24 and Hsp70 were detected by immunohistochemistry and their correlation was analyzed. The expression levels of CD24 mRNA and protein were examined using qRT-PCR and western blotting in SPCA1, A549, H1975, and H1650 cell lines. A CD24-overexpressing cell model was established. The interaction between CD24 and Hsp70 was verified by co-immunoprecipitation and western blotting. CD24 and Hsp70 expression were significantly higher in lung cancer tissues than in adjacent tissues (CD24: p=0.008; Hsp70: p<0.001). CD24 protein expression showed a positive correlation with lymph node metastasis, TNM stage, and vascular cancer thrombus. Hsp70 protein expression showed a positive correlation with differentiation, lymph node metastasis, and TNM stage. CD24 and Hsp70 high expression were also correlated with poor survival. The positive co-expression rate of CD24 and Hsp70 in lung cancer tissues was 52.7% (49/93). CD24 and Hsp70 expression in lung cancer were positively correlated (r=0.368, p<0.001), and co-immunoprecipitation was verified that both endogenous and exogenous CD24 co-precipitated with Hsp70 directly or indirectly. When Hsp70 inhibitor VER15508 was added to A549 cells, Hsp70 and CD24 protein expression were significantly decreased. The present study demonstrated that CD24 and Hsp70 were highly expressed in lung cancer tissues, and associated with invasion, metastasis, and poor survival. Hsp70 may regulate CD24 expression. Co-expression of CD24 and Hsp70 may be a prognostic biomarker for lung cancer.


Pinched-flow hydrodynamic stretching of single-cells.
Reorganization of cytoskeletal networks, condensation and decondensation of chromatin, and other whole cell structural changes often accompany changes in cell state and can reflect underlying disease processes. As such, the observable mechanical properties, or mechanophenotype, which is closely linked to intracellular architecture, can be a useful label-free biomarker of disease. In order to make use of this biomarker, a tool to measure cell mechanical properties should accurately characterize clinical specimens that consist of heterogeneous cell populations or contain small diseased subpopulations. Because of the heterogeneity and potential for rare populations in clinical samples, single-cell, high-throughput assays are ideally suited. Hydrodynamic stretching has recently emerged as a powerful method for carrying out mechanical phenotyping. Importantly, this method operates independently of molecular probes, reducing cost and sample preparation time, and yields information-rich signatures of cell populations through significant image analysis automation, promoting more widespread adoption. In this work, we present an alternative mode of hydrodynamic stretching where inertially-focused cells are squeezed in flow by perpendicular high-speed pinch flows that are extracted from the single inputted cell suspension. The pinched-flow stretching method reveals expected differences in cell deformability in two model systems. Furthermore, hydraulic circuit design is used to tune stretching forces and carry out multiple stretching modes (pinched-flow and extensional) in the same microfluidic channel with a single fluid input. The ability to create a self-sheathing flow from a single input solution should have general utility for other cytometry systems and the pinched-flow design enables an order of magnitude higher throughput (65,000 cells s(-1)) compared to our previously reported deformability cytometry method, which will be especially useful for identification of rare cell populations in clinical body fluids in the future.


Digital stethoscope: technology update.
Cardiovascular disease (CVD) is recognized as the leading cause of mortality throughout the world. About one-third of global mortality is attributable to CVD. In addition to clinical presentation, specific clinical exam findings can assist in treating and preventing CVD. CVD may initially manifest as pulmonary pathology, and thus, accurate cardiopulmonary auscultation is paramount to establishing accurate diagnosis. One of the most powerful tools available to physicians is the stethoscope. The stethoscope first emerged in the year 1818, invented by a French physician, René Laennec. Since then, the initial modest monaural wooden tube has evolved into a sophisticated digital device. This paper provides an analysis of the evolution of the stethoscope as well as highlights the advancement made by the modern digital stethoscope including the application of this tool in advancing care for patients suffering from CVD.


HYCUD: a computational tool for prediction of effective rotational correlation time in flexible proteins.
A large fraction of eukaryotic proteins contain unstructured tails or linkers. The presence of flexible regions allows these systems to experience a high level of mobility facilitating their biological function. The complex nature of protein rotation in such flexible modular systems precludes a straightforward application of hydrodynamic methods to calculate their rotational motional properties. We describe the workflow of HYdrodynamic CoUpling of Domains (HYCUD), a program for prediction of effective rotational correlation times in multidomain proteins. The usage of HYCUD is demonstrated by its application to the ribosomal protein L7/L12. Rotational correlation times predicted by HYCUD might be used to detect molecular switch events mediated by disorder-order transitions in interdomain linkers. The source code and documentation are available at www.mpibpc.mpg.de/106144/software. firstname.lastname@example.com or firstname.lastname@example.com Supplementary material is available at Bioinformatics online.


Plant cell-made protein antigens for induction of Oral tolerance.
The gut associated lymphoid tissue has effective mechanisms in place to maintain tolerance to food antigens. These can be exploited to induce antigen-specific tolerance for the prevention and treatment of autoimmune diseases and severe allergies and to prevent serious immune responses in protein replacement therapies for genetic diseases. An oral tolerance approach for the prevention of peanut allergy in infants proved highly efficacious and advances in treatment of peanut allergy have brought forth an oral immunotherapy drug that is currently awaiting FDA approval. Several other protein antigens made in plant cells are in clinical development. Plant cell-made proteins are protected in the stomach from acids and enzymes after their oral delivery because of bioencapsulation within plant cell wall, but are released to the immune system upon digestion by gut microbes. Utilization of fusion protein technologies facilitates their delivery to the immune system, oral tolerance induction at low antigen doses, resulting in efficient induction of FoxP3+ and latency-associated peptide (LAP)+ regulatory T cells that express immune suppressive cytokines such as IL-10. LAP and IL-10 expression represent potential biomarkers for plant-based oral tolerance. Efficacy studies in hemophilia dogs support clinical development of oral delivery of bioencapsulated antigens to prevent anti-drug antibody formation. Production of clinical grade materials in cGMP facilities, stability of antigens in lyophilized plant cells for several years when stored at ambient temperature, efficacy of oral delivery of human doses in large animal models and lack of toxicity augur well for clinical advancement of this novel drug delivery concept.


Long-Term Impacts of Foetal Malnutrition Followed by Early Postnatal Obesity on Fat Distribution Pattern and Metabolic Adaptability in Adult Sheep.
We aimed to investigate whether over- versus undernutrition in late foetal life combined with obesity development in early postnatal life have differential implications for fat distribution and metabolic adaptability in adulthood. Twin-pregnant ewes were fed NORM (100% of daily energy and protein requirements), LOW (50% of NORM) or HIGH (150%/110% of energy/protein requirements) diets during the last trimester. Postnatally, twin-lambs received obesogenic (HCHF) or moderate (CONV) diets until 6 months of age, and a moderate (obesity correcting) diet thereafter. At 2½ years of age (adulthood), plasma metabolite profiles during fasting, glucose, insulin and propionate (in fed and fasted states) tolerance tests were examined. Organ weights were determined at autopsy. Early obesity development was associated with lack of expansion of perirenal, but not other adipose tissues from adolescence to adulthood, resulting in 10% unit increased proportion of mesenteric of intra-abdominal fat. Prenatal undernutrition had a similar but much less pronounced effect. Across tolerance tests, LOW-HCHF sheep had highest plasma levels of cholesterol, urea-nitrogen, creatinine, and lactate. Sex specific differences were observed, particularly with respect to fat deposition, but direction of responses to early nutrition impacts were similar. However, prenatal undernutrition induced greater metabolic alterations in adult females than males. Foetal undernutrition, but not overnutrition, predisposed for adult hypercholesterolaemia, hyperureaemia, hypercreatinaemia and hyperlactataemia, which became manifested only in combination with early obesity development. Perirenal expandability may play a special role in this context. Differential nutrition recommendations may be advisable for individuals with low versus high birth weights.


Health Conditions and Memory Performance: a study with older adult women.
to verify the correlation between health condition and memory performance of older adult women in the community. Analytical cross-sectional study developed with 28 older adult women living in Cuiabá-MT. They answered the Mini-Mental State Examination (MMSE), and a shortened Geriatric Depression Scale (GDS-15) to screen for dementia and depression symptoms. Memory skills were assessed through Rey Auditory Verbal Learning Test (RAVLT). The mean age was 66.36 years and 75% of the participants had educational level higher than 7 years. The MMSE mean score was 28.45. The correlations found were: educational level and immediate memory (r = 0.49; p = 0.008); delayed recall and immediate memory (r = 0.71; p < 0.001); memory recognition and immediate memory (r = 0.43; p = 0.021) and recognition memory with delayed recall (r = 0.47; p = 0.012). High scores in the MMSE and a satisfactory health perception among the participants were evident. There was no correlation between memory performance and health perception.


Gaze as depicted in Vermeer's Girl with a Pearl Earring.
Ten observers viewed Vermeer's painting Girl With a Pearl Earring and estimated her direction of gaze when her eyes were exposed together and separately. The observers also viewed the painting when her eyes were digitally replaced by those of a real person. The authors found that Vermeer painted the girl's eyes with a precision that was near the limits of visual acuity. Also, Vermeer included 3 gaze illusions, none of which researchers have documented as known in Vermeer's time: (a) when a model whose head is turned to one side gazes at an observer, there is an illusion that she is looking to the side of the observer away from the direction of her head turn; (b) when a model's eyes are viewed separately, they appear to gaze outward relative to each other; and (c) when both of a model's eyes are viewed, the perceived direction of gaze follows that of the nearer eye.