Distribution and repeatability of anterior pituitary responses to GnRH and relationship of response classification to the postpartum anovulatory interval of beef cows.
Our objectives were to investigate the phenotypic variation in anterior pituitary responsiveness to GnRH (100 microg, i.v.) of beef cows between d 5 and 8 postpartum, estimate repeatability, and determine the relationship between response classification and duration of the postpartum anovulatory interval (PPI). Brahman x Hereford (F1) cows (n = 137) and primiparous heifers (n = 58) were evaluated. Response classifications (Class) included peak LH (Low, Intermediate, or High; Class I) and time to peak LH (Early, 10 to 30 min or Late, 60 to 120 min; Class II). The independent effects of Class I and II on PPI were determined in 145 of 195 cows through twice-weekly serum samples analyzed for progesterone. For Class I, pituitary responses to GnRH approximated a normal distribution and, by definition, differed (P < .001) in magnitudes of peak LH and area under the curve (AUC). For Class II, 111 and 84 cows exhibited early and late peaks, respectively; mean AUC was greater (P < .05) in cows exhibiting late compared with early peaks. Pretreatment LH (P < .01) and estradiol-17beta (P < .004) influenced responses in one or both response classes. Pluriparous cows had shorter (P < .035) PPI than primiparous cows. Class I did not influence the duration of the PPI; however, in Class II, cows with late peaks exhibited an average PPI that was 8 d shorter (P < .025) than in those with an early peak. To estimate repeatability of pituitary responses, 18 classified cows were subsequently rechallenged with GnRH at d 170 of gestation and at the next postpartum period. Although means for each of these challenges differed (P < .05) throughout in both Classes I and II, the small sample size used to make the estimate failed to yield significant (P > .10) interclass correlations. Nevertheless, overall results provide evidence that variability in individual pituitary responses to GnRH could be targeted as a selection marker to improve reproduction.


[Spatial heterogeneity in body condition of small yellow croaker in Yellow Sea and East China Sea based on mixed-effects model and quantile regression analysis].
By using the 2008-2010 investigation data about the body condition of small yellow croaker in the offshore waters of southern Yellow Sea (SYS), open waters of northern East China Sea (NECS), and offshore waters of middle East China Sea (MECS), this paper analyzed the spatial heterogeneity of body length-body mass of juvenile and adult small yellow croakers by the statistical approaches of mean regression model and quantile regression model. The results showed that the residual standard errors from the analysis of covariance (ANCOVA) and the linear mixed-effects model were similar, and those from the simple linear regression were the highest. For the juvenile small yellow croakers, their mean body mass in SYS and NECS estimated by the mixed-effects mean regression model was higher than the overall average mass across the three regions, while the mean body mass in MECS was below the overall average. For the adult small yellow croakers, their mean body mass in NECS was higher than the overall average, while the mean body mass in SYS and MECS was below the overall average. The results from quantile regression indicated the substantial differences in the allometric relationships of juvenile small yellow croakers between SYS, NECS, and MECS, with the estimated mean exponent of the allometric relationship in SYS being 2.85, and the interquartile range being from 2.63 to 2.96, which indicated the heterogeneity of body form. The results from ANCOVA showed that the allometric body length-body mass relationships were significantly different between the 25th and 75th percentile exponent values (F=6.38, df=1737, P<0.01) and the 25th percentile and median exponent values (F=2.35, df=1737, P=0.039). The relationship was marginally different between the median and 75th percentile exponent values (F=2.21, df=1737, P=0.051). The estimated body length-body mass exponent of adult small yellow croakers in SYS was 3.01 (10th and 95th percentiles = 2.77 and 3.1, respectively). The estimated body length-body mass relationships were significantly different from the lower and upper quantiles of the exponent (F=3.31, df=2793, P=0.01) and the median and upper quantiles (F=3.56, df=2793, P<0.01), while no significant difference was observed between the lower and median quantiles (F=0.98, df=2793, P=0.43).


Structural basis for the binding of compatible solutes by ProX from the hyperthermophilic archaeon Archaeoglobus fulgidus.
Compatible solutes such as glycine betaine and proline betaine serve as protein stabilizers because of their preferential exclusion from protein surfaces. To use extracellular sources of this class of compounds as osmo-, cryo-, or thermoprotectants, Bacteria and Archaea have developed high affinity uptake systems of the ATP-binding cassette type. These transport systems require periplasmic- or extracellular-binding proteins that are able to bind the transported substance with high affinity. Therefore, binding proteins that bind compatible solutes have to avoid the exclusion of their ligands within the binding pocket. In the present study we addressed the question to how compatible solutes can be effectively bound by a protein at temperatures around 83 degrees C as this is done by the ligand-binding protein ProX from the hyperthermophilic archaeon Archaeoglobus fulgidus. We solved the structures of ProX without ligand and in complex with both of its natural ligands glycine betaine and proline betaine, as well as in complex with the artificial ligand trimethylammonium. Cation-pi interactions and non-classical hydrogen bonds between four tyrosine residues, a main chain carbonyl oxygen, and the ligand have been identified to be the key determinants in binding the quaternary amines of the three investigated ligands. The comparison of the ligand binding sites of ProX from A. fulgidus and the recently solved structure of ProX from Escherichia coli revealed a very similar solution for the problem of compatible solute binding, although both proteins share only a low degree of sequence identity. The residues involved in ligand binding are functionally equivalent but not conserved in the primary sequence.


Coevolution of a virus-alga system.
Plectonema boryanum, a filamentous blue-green alga, was cloned and then allowed to reach a steady state in a quasi-continuous culture in the presence of the algal virus, LPP-1. The culture was maintained for 3.5-month period during which time at least four distinct culture lysings were evident. After the fourth lysis the culture reached a steady-state level which was identical in its algal concentration to the preinfection level. Upon testing the characteristics of the evolved alga and virus variants, the following was determined: cell variants resistant to both the original virus and the derived virus had evolved, and there was no evidence of lysogeny present amony these cells. The evolved virus strains still grew on the parental algal strain, though with altered plaque morphology. Furthermore, they were antigenically similar to the parental virus, and showed no significant difference in adsorption rate or growth characteristics on parental cells. However, a low-grade chronic viral infection persisted in the culture. Rapid re-establishment of a dense, stable culture is apparantly the normal laboratory response of a procaryotic cell-virus system.


Characterization of Ser338 phosphorylation for Raf-1 activation.
Raf kinases are essential for regulating cell proliferation, survival, and tumorigenesis. However, the mechanisms by which Raf is activated are still incompletely understood. Phosphorylation plays a critical role in Raf activation in response to mitogens. The present study characterizes phosphorylation of Ser338, a crucial event for Raf-1 activation. Here we report that mutation of Lys375 to Met diminishes phosphorylation of Ser338 on both wild type Raf-1 in cells treated with epidermal growth factor (EGF) or 12-O-tetradecanoylphorbol-13-acetate (TPA) and a constitutively active mutant in which Tyr340/Tyr341 are replaced by 2 aspartic acids, a conserved substitution present in natural B-Raf. The loss of Ser338 phosphorylation in these Raf mutants is not engendered by a mutation-induced conformational change, inasmuch as mutation of another site (Ser471 to Ala) in the activation segment also abolishes Ser338 phosphorylation, whereas both the kinase-dead mutants of Raf-1 are phosphorylated well by active Pak1. Furthermore, our data demonstrate that EGF-stimulated phosphorylation of Ser338 is inhibited by Sorafenib, a Raf kinase inhibitor, but not by the MEK inhibitor U0126. Interestingly, a kinase-dead mutation and Sorafenib also markedly reduce phosphorylation of Ser445 on B-Raf, a site equivalent to Raf-1 Ser338. Finally, our data reveal that Ser338 is phosphorylated on inactive Raf-1 by an active mutant of Raf-1 when they are dimerized in cells and that artificial dimerization of Raf-1 causes Ser338 phosphorylation, accompanied by activation of ERK1/2. Altogether, our data suggest that Ser338 on Raf-1 is autophosphorylated in response to mitogens.


Sclerosing stromal tumor of the ovary. A case report and further observations.
A case of sclerosing stromal tumor of the ovary in a 16-year-old girl is described. The lesion had the characteristic mixed pseudolobular pattern with cellular areas separated by edematous stroma. A prominent, additional feature was the presence of partially or completely obstructed vessels at the hilus. It is proposed that these vascular changes may be the cause of the edema in the tumor.


Gold nanoparticle assemblies stabilized by bis(phthalocyaninato)lanthanide(III) complexes through van der Waals interactions.
Gold nanoparticle assemblies possess diverse application potential, ranging from industrial nanotechnology to medical biotechnology. Because the structures and properties of assemblies are directly affected by the stabilization mechanism between the organic molecules serving as protecting ligands and the gold nanoparticle surface, it is crucial to find and investigate new stabilization mechanisms. Here, we report that π-conjugated phthalocyanine rings can serve as stabilizing ligands for gold nanoparticles. Bis(phthalocyaninato)lutetium(III) (LuPc2) or bis(phthalocyaninato)terbium(III) (TbPc2), even though complex, do not have specific binding units and stabilize gold nanoparticles through van der Waals interaction between parallel adsorbed phthalocyanine ligands and the gold nanoparticle surface. AC magnetic measurements and the electron-transport properties of the assemblies give direct evidence that the phthalocyanines are isolated from each other. Each nanoparticle shows weak electronic coupling despite the short internanoparticle distance (~1 nm), suggesting Efros-Shklovskii-type variable-range hopping and collective single-electron tunnelling behaviours.


Acute acneiform eruption induced by interferon beta-1b during treatment for multiple sclerosis.
Multiple sclerosis is an inflammatory demyelinating disease of presumed autoimmune origin that affects the central nervous system. The main form of therapy is based on the use of immunomodulators such as interferon beta, which are usually well tolerated. Skin manifestations resulting from treatment with interferon beta-1b consist principally of reactions at the site of subcutaneous application of the drug. The present case report describes a female patient who developed an acneiform eruption resulting from treatment with interferon beta-1b.


[Evaluation of motherś knowledge about oral rehydration therapy and sodium concentration in homemade sugar salt solutions]
OBJECTIVE: To evaluate motherś knowledge about oral rehydration therapy and to determine the concentration of sodium in sugar salt solutions prepared by the interviewed mothers. METHODS: Cross-sectional study of low-income families from the suburbs of Natal, a city in the state of Rio Grande do Norte. One hundred and thirty six households with children younger than 5 years were visited. The information on when to use the solution, how to administrate it and how to prepare it was provided by the interviewed mothers. The concentration of sodium was determined in 100 solutions prepared by those mothers. RESULTS: Only 9.0% of mothers had appropriate knowledge about when to use the solution and 21.0% knew how to administer it correctly. The error rates regarding the measurement of ingredients were: 2.5% for salt, 22.2% for water and 43.2% for sugar, according to the answers given by 81 mothers, who used a double-ended spoon made of plastic to measure quantities. In fifty percent of the analyzed samples, sodium was within safe limits (30 mmol/l to 100 mmol/l). Those considered dangerous (>100 mmol/l) or inefficient solutions (<30 mmol/l) reached the percentage of 47.0% and 3.0%, respectively. Out of the 69 solutions prepared with the double-ended spoon, 63.8% were found appropriate in terms of sodium concentration, while only 19.3% of the 31 solutions prepared using other measuring utensils were within acceptable limits (P=0.0001). CONCLUSION: Motherś knowledge about oral rehydration therapy was considered poor. Moreover, the concentration of sodium in sugar salt solutions seldom remains within safe limits, particularly those solutions in which the double-ended spoon was not used for preparation. These conclusions should be considered in future educational programs on oral rehydration.


Pediatric Cancer Variant Pathogenicity Information Exchange (PeCanPIE): a cloud-based platform for curating and classifying germline variants.
Variant interpretation in the era of massively parallel sequencing is challenging. Although many resources and guidelines are available to assist with this task, few integrated end-to-end tools exist. Here, we present the Pediatric Cancer Variant Pathogenicity Information Exchange (PeCanPIE), a web- and cloud-based platform for annotation, identification, and classification of variations in known or putative disease genes. Starting from a set of variants in variant call format (VCF), variants are annotated, ranked by putative pathogenicity, and presented for formal classification using a decision-support interface based on published guidelines from the American College of Medical Genetics and Genomics (ACMG). The system can accept files containing millions of variants and handle single-nucleotide variants (SNVs), simple insertions/deletions (indels), multiple-nucleotide variants (MNVs), and complex substitutions. PeCanPIE has been applied to classify variant pathogenicity in cancer predisposition genes in two large-scale investigations involving >4000 pediatric cancer patients and serves as a repository for the expert-reviewed results. PeCanPIE was originally developed for pediatric cancer but can be easily extended for use for nonpediatric cancers and noncancer genetic diseases. Although PeCanPIE's web-based interface was designed to be accessible to non-bioinformaticians, its back-end pipelines may also be run independently on the cloud, facilitating direct integration and broader adoption. PeCanPIE is publicly available and free for research use.


Biosynthesis of chitin by particulate fractions from Cunnighamella elegans.
The enzyme chitin synthetase (UDP-acetylaminodeoxyglucosyl transferase, EC 188.8.131.52) in Cunninghamella elegans has been investigated. The enzyme was present in the microsomal, cell wall, mitochondrial and the soluble cytoplasmic fraction of the mycelium, with the former having the highest specific activity. The properties of the enzyme in this fraction were investigated; the Km for UDP GlcNAc was 1.23 mM and 2.08 mM GlcNAc in the presence of 1 mM UDP GlcNAc. The temperature optimum was between 26 degrees and 29 degrees C and maximal activity was at pH 6.25. Mg++ ions had no effect on chitin synthesis, but soluble chitodextrins inhibited the enzyme. The production of chitin synthetase was correlated with the growth of the fungus, maximum activity being found during the late exponential phase of growth. Chitin was confirmed as the sole product of enzyme action, by digestion with chitinase.


Lower urinary tract symptoms and erectile dysfunction: co-morbidity or typical "Aging Male" symptoms? Results of the "Cologne Male Survey".
Due to demographic developments in industrial nations, age-correlated diseases are becoming more important. From many epidemiological studies we know that the prevalence of benign prostatic hyperplasia (BPH) and the loss of erectile function (= erectile dysfunction or ED) increase with advancing age. Are these two illnesses related or/and independent? We mailed our newly developed and validated questionnaire on male erectile dysfunction (KEED), as well as a set of questions pertaining to voiding problems (IPSS), to a representative population sample of 8000 men from 30 to 80 years of age residing in the city of Cologne. The responses included 4489 evaluable replies (56.1%). The response rates in the different age groups ranged from 41 to 61%. The mean age of the men who answered was 51.8 years. The overall prevalence of ED was 19.2% (n=862), with a steep age-related increase from 2 to 53%. Furthermore, 31.2% (n=1957) of all men complained of lower urinary tract symptoms (LUTS), the prevalence and the intensity of which increased with age. Interestingly, a high co-morbidity was found between ED and voiding problems. Prevalence of LUTS in men suffering from ED was about 72.2% (n=621) vs. 37.7% (1367) in men with normal erections. The odds ratio was evaluated with 2.11. The trivariate analysis showed that the occurrence of LUTS can be considered as an age-independent risk factor for the development of ED (p<0.001). Even though the pathogenetic relationship between LUTS and ED is not yet completely understood, one has to postulate a direct association between these two typical symptom complexes in the aging male.


Vesicular zinc regulates the Ca2+ sensitivity of a subpopulation of presynaptic vesicles at hippocampal mossy fiber terminals.
Synaptic vesicles segregate into functionally diverse subpopulations within presynaptic terminals, yet there is no information about how this may occur. Here we demonstrate that a distinct subgroup of vesicles within individual glutamatergic, mossy fiber terminals contain vesicular zinc that is critical for the rapid release of a subgroup of synaptic vesicles during increased activity in mice. In particular, vesicular zinc dictates the Ca(2+) sensitivity of release during high-frequency firing. Intense synaptic activity alters the subcellular distribution of zinc in presynaptic terminals and decreases the number of zinc-containing vesicles. Zinc staining also appears in endosomes, an observation that is consistent with the preferential replenishment of zinc-enriched vesicles by bulk endocytosis. We propose that functionally diverse vesicle pools with unique membrane protein composition support different modes of transmission and are generated via distinct recycling pathways.


[Morphology and anatomy of the fruit development of Ocotea puberula (Rich.) Nees and Nectandra megapotamica (Spreng.) Mez (Lauraceae)].
The morphology and anatomy of the fruit development of Ocotee puberula (Rich.) Nees and Nectandra megapotamica (Spreng.) Mez (Lauraceae) was studied in flowers and fruits collected in secondary forests of Maringa, Parana State, Brazil. The fruits of the species are drupes, with epidermic exocarp, parenchymatous mesocarp and endocarp which consists of macrosclereids. The origin of endocarp is the internal epidermis of ovary. The exalbuminous seeds develop from the anatropous ovules and they have testa and tegmen with parenchymatous cells frequently compressed. In the hilum region these teguments show sciereids in radial arrangement. The straight embryo has thick cotyledons which contain reserve substances such as starch and oil. The plumule and hypocotyl-radicle axis are reduced.


A Qualitative Analysis to Understand Perception about Medication-Related Problems among Older Minority Adults in a Historically Black Community.
Older adults taking multiple chronic medications experience an increased risk of adverse drug events and other medication-related problems (MRP). Most current literature on medication management involves researcher-driven intervention, yet few studies investigate patients' understanding of MRP in a diverse community setting. This report investigates patients' perception of MRP and patient-centered strategies among a cohort of the older adult group in a historically Black urban community. The study design is qualitative using structured open-ended questions in a multidisciplinary patient-centered focus group. Patients (age 65 years or older) taking seven or more medications were recruited. The group comprises patients, caregivers, pharmacists, health educators, a physician, and a nurse. Recordings of the group discussion are transcribed verbatim and analyzed using thematic content analysis and categorized by codes developed from the social-ecological model. The group reports patient-provider relationships, previous experience, fear of side effects played important roles in medication adherence. There is an unmet need for medication management education and tools to organize complex medication lists from multiple providers. This study provides important insights into MRP experienced by minority older adults and provided researchers with potential strategies for future interventions.


Analysis of the nonlinear dynamics of daily broiler growth and feed intake.
Daily BW velocity (BWV) and acceleration (BWA) of individual birds have been demonstrated to be oscillatory. Daily feed intake velocity (FIV) and acceleration (FIA) were hypothesized to be oscillatory and to have a positive relationship with BWV and BWA, respectively. Forty-eight male broiler chicks were individually caged and provided a commercial starter feed ad libitum for 49 d. BW and feed intake (FI) were measured daily. Experiment 1 confirmed that, on a daily basis, BWV, BWA, FIV, and FIA were oscillatory. There was a positive correlation between BW and FI, BWV and FIV, and BWA and FIA. A Kohonen neural network (KNN) clustered BWV and FIV into two and three sequential phases. BWA and FIA analysis did not make definitive clusters. In experiment 2, it was hypothesized that correlation between BWV and FIV would increase with feeding of grower and finisher rations. It was hypothesized that KNN three phase clusters may provide more biologically ideal times of ration change (TORC) for starter, grower, and finisher rations. For 49 d, five treatments, nine birds per treatment, were fed starter, grower, and finisher rations singly or together with dietary changes according to an industry or KNN-determined TORC. Evaluation was made of BW, FI, and carcass characteristics. No significant mean differences were found. Compared to the industry group, the KNN group demonstrated significantly improved uniformity (i.e., smaller SD) of BW (bled out), FI, dressing percentage, and some of the carcass characteristics. Differences between KNN and industry TORC results might have been related to the length of time the birds were fed the starter, grower, and finisher diets.


Allosteric activation mechanism of the alpha 1 beta 2 gamma 2 gamma-aminobutyric acid type A receptor revealed by mutation of the conserved M2 leucine.
A conserved leucine residue in the midpoint of the second transmembrane domain (M2) of the ligand-activated ion channel family has been proposed to play an important role in receptor activation. In this study, we assessed the importance of this leucine in the activation of rat alpha 1 beta 2 gamma 2 GABA receptors expressed in Xenopus laevis oocytes by site-directed mutagenesis and two-electrode voltage clamp. The hydrophobic conserved M2 leucines in alpha1(L263), beta2(L259), and gamma 2(L274) subunits were mutated to the hydrophilic amino acid residue serine and coexpressed in all possible combinations with their wild-type and/or mutant counterparts. The mutation in any one subunit decreased the EC(50) and created spontaneous openings that were blocked by picrotoxin and, surprisingly, by the competitive antagonist bicuculline. The magnitudes of the shifts in GABA EC(50) and picrotoxin IC(50) as well as the degree of spontaneous openings were all correlated with the number of subunits carrying the leucine mutation. Simultaneous mutation of the GABA binding site (beta 2Y157S; increased the EC(50)) and the conserved M2 leucine (beta 2L259S; decreased the EC(50)) produced receptors with the predicted intermediate agonist sensitivity, indicating the two mutations affect binding and gating independently. The results are discussed in light of a proposed allosteric activation mechanism.


Preconditioning with Foam-mediated External Suction on Flap Microvasculature and Perfusion in a Rodent Model.
Foam-mediated external suction (FMES) has previously shown to improve tissue microcirculation. We hypothesized that preconditioning fasciocutaneous perforator flaps with FMES would augment perfusion and demonstrate greater capillary recruitment. Gluteal perforator flaps were designed on sixteen 400 g rats. Continuous FMES at -125 mm Hg was applied on one side (intervention) to precondition tissue for 5 days, with the contralateral side as a paired control. In group A, we assessed changes following pretreatment, after surgery, and 7 days postprocedure, and in group B, we evaluated changes during preconditioning alone. In group A (N = 8), control and intervention flaps were assessed using laser-assisted indocyanine green fluorescence angiography. In group B, flap regions were assessed using 4-dimensional computed tomographic angiography. All flaps were analyzed for microvessel density using micro-computed tomography and histological assessment using hematoxylin and eosin and CD3 immunohistochemistry. Thirty-two flaps were included in this study (N = 16 intervention and matched controls). Four-dimensional computed tomographic angiography demonstrated 17% greater tissue perfusion in preconditioned flaps (mean, 78.7 HU; SD, 8.8) versus controls (mean, 67.3 HU; SD, 15.7; P < 0.01). Laser-assisted indocyanine green fluorescence angiography showed a 30% higher mean absolute intensity in preconditioned flaps versus controls (P < 0.01). Postsurgery mean absolute intensity in preconditioned flaps remained 21% higher than in controls (P = 0.03). Preconditioned flaps demonstrated a 2-fold increase in mean vessel volume of 9.1 mm3 (SD, 7) versus 4.5 mm3 (SD, 3) in controls (P = 0.04); there was a 33% higher mean area fraction of CD31 in preconditioned flaps, 3.9% (SD, 3) versus 2.9% (SD, 3) in controls (P = 0.03). FMES preconditioning has the potential to augment vascularity of tissue for flap harvest; however, further experimental studies are required to optimize strategies and evaluate long-term effects for clinical applications.


Localization of cell surface sites involved in fibronectin fibrillogenesis.
Fibronectin binding sites on cultured human fibroblasts were localized by high voltage electron microscopy using either 5- or 18-nm colloidal gold beads (Au5 or Au18) bound to intact fibronectin, the 70-kD amino-terminal fragment of fibronectin that blocks incorporation of exogenous fibronectin into extracellular matrix, or 160-180-kD fragments of fibronectin with cell adhesion and heparin-binding activities. Binding sites for Au18-fibronectin on the cell surface were localized to specific regions along the edge of the fibroblast and on retraction fibers. Au18-fibronectin complexes at these sites were initially localized in clusters that co-aligned with intracellular microfilament bundles. With longer incubations, Au18-fibronectin complexes were arranged into long fibrillar networks on the cell surface and in the extracellular space. The appearance of Au18-fibronectin in these fibrillar networks and disappearance of clusters of Au18-fibronectin suggest that Au18-fibronectin complexes are arranged into matrix at specific regions of the cell surface. Au18-70-kD fragment complexes initially had a similar distribution to Au18-fibronectin complexes. With longer incubations, Au18-70-kD fragment complexes were found in long linear arrangements on the cell surface. Double labeling experiments using Au18-70-kD fragment and Au5-160-180-kD fragments showed that the 70-kD fragment and the 160-180-kD fragments bind to different regions of the cell.


Specific mutation of a regulatory site within the ATP-binding region of simian virus 40 large T antigen.
In an attempt to distinguish simian virus 40 (SV40) large T antigen (T) binding to ATP from hydrolysis, specific mutations were made in the ATP-binding site of T according to our model for the site (M. K. Bradley, T. F. Smith, R. H. Lathrop, D. M. Livingston, and T. A. Webster, Proc. Natl. Acad. Sci. USA 84:4026-4030, 1987). Two acidic residues predicted to make contact with the magnesium phosphate were changed to alanines. The mutated T gene was completely defective for viral DNA synthesis and for virion production, and it was dominant defective for viral DNA replication. The defective T gene encoded a stable product (2905T) that oncogenically transformed mouse cell lines. 2905T, immunoprecipitated from transformed-cell extracts, bound SV40 origin DNA specifically and, surprisingly, it was active as an ATPase. A recombinant baculovirus was constructed for the production and purification of the mutant protein for detailed biochemical analyses. 2905T had only 10% of the ATPase and helicase of wild-type T. The Km of 2905T for ATP in ATPase assays was the same as the Km of wild-type T. ATP activated the ATPase activity of wild-type T, but not of 2905T. As tested by gel bandshift assay, 2905T bound to SV40 origin DNA and to individual sites I and II with affinities similar to that of the wild type. However, ATP did not modulate the DNA-binding activity of mutant T to site II. Therefore, this mutation in the ATP-binding site in T resulted in defects in the interaction between the protein and ATP that appeared to be responsible for the determination of the active state of T for DNA binding versus ATPase.


Short-term long chain omega3 diet protects from neuroinflammatory processes and memory impairment in aged mice.
Regular consumption of food enriched in omega3 polyunsaturated fatty acids (ω3 PUFAs) has been shown to reduce risk of cognitive decline in elderly, and possibly development of Alzheimer's disease. Docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) are the most likely active components of ω3-rich PUFAs diets in the brain. We therefore hypothesized that exposing mice to a DHA and EPA enriched diet may reduce neuroinflammation and protect against memory impairment in aged mice. For this purpose, mice were exposed to a control diet throughout life and were further submitted to a diet enriched in EPA and DHA during 2 additional months. Cytokine expression together with a thorough analysis of astrocytes morphology assessed by a 3D reconstruction was measured in the hippocampus of young (3-month-old) and aged (22-month-old) mice. In addition, the effects of EPA and DHA on spatial memory and associated Fos activation in the hippocampus were assessed. We showed that a 2-month EPA/DHA treatment increased these long-chain ω3 PUFAs in the brain, prevented cytokines expression and astrocytes morphology changes in the hippocampus and restored spatial memory deficits and Fos-associated activation in the hippocampus of aged mice. Collectively, these data indicated that diet-induced accumulation of EPA and DHA in the brain protects against neuroinflammation and cognitive impairment linked to aging, further reinforcing the idea that increased EPA and DHA intake may provide protection to the brain of aged subjects.


B vitamin supplementation improves cognitive function in the middle aged and elderly with hyperhomocysteinemia.
An intervention study was performed to determine if supplement containing folic acid, vitamin B6, and vitamin B12 could improve cognitive function and lower homocysteine in middle-aged and elderly patients with hyperhomocysteinemia. One hundred and four participants with hyperhomocysteinemia were recruited in Tianjin, China, aged 55-94 years old. Fifty-seven individuals with hyperhomocysteinemia were included in the intervention group (vitamin B group, which received 800 µg/day of folate, with 10 mg of vitamin B6 and 25 µg of vitamin B12) and 47 patients in the placebo group. The endpoint was the improvement in cognitive function as evaluated by Basic Cognitive Aptitude Tests (BCATs). All parameters were measured before and after the treatment period of 14 weeks. The BCAT total score and four sub-tests scores (digit copy, Chinese character rotation, digital working memory, and recognition of meaningless figure) of BCAT at 14 weeks significantly increased only for the vitamin B group. Serum total homocysteine (tHcy) levels significantly decreased in the intervention group, while serum concentrations of folate, vitamin B6, and vitamin B12 significantly increased in the intervention group. The results demonstrated that supplement containing folate, vitamin B6, and vitamin B12 in middle-aged and elderly patients with hyperhomocysteinemia could improve their cognitive function partly and reduce serum tHcy levels.


Molecular barriers to processes of genetic reprogramming and cell transformation.
Genetic reprogramming by ectopic expression of transcription factor genes induces the pluripotent state in somatic cells. This technology provides an opportunity to establish pluripotent stem cells for each person, as well as to get better understanding of epigenetic mechanisms controlling cell state. Interestingly, some of the molecular processes that accompany somatic cell reprogramming in vitro are also characteristic for tumor manifestation. Thus, similar "molecular barriers" that control the stability of epigenetic state exist for both processes of pluripotency induction and malignant transformation. The reprogramming of tumor cells is interesting in two aspects: first, it will determine the contribution of epigenetic changes in carcinogenesis; second, it gives an approach to evaluate tumor stem cells that are supposed to form the entire cell mass of the tumor. This review discusses the key stages of genetic reprogramming, the similarity and difference between the reprogramming process and malignant transformation.


Association between interruptions in medicaid coverage and use of inpatient psychiatric services.
Persons with schizophrenia are heavy and persistent users of Medicaid services. Interruptions in their Medicaid coverage may have serious consequences for the mental health of these individuals and their subsequent use of mental health services. This study sought to determine the impact of interruptions in Medicaid coverage on the use of Medicaid-reimbursed inpatient psychiatric services over a four-year period. Inpatient Medicaid claims and eligibility files for 1,830 Medicaid beneficiaries with schizophrenia in Utah from December 1990 to December 1994 were used to identify differences in hospital admissions and total number of days in a hospital associated with interrupted Medicaid coverage. Of the 1,830 Medicaid beneficiaries identified, 1,463 experienced continuous Medicaid eligibility, and 367 had interruptions in their eligibility. Interruptions in Medicaid coverage were associated with an average of.63 more psychiatric hospitalizations per beneficiary over the four-year period, representing an 86 percent higher hospital admission rate. This increase appeared to be largely due to a subset of persons who have much higher hospitalization rates after an interruption in Medicaid coverage. Interruptions in Medicaid coverage were associated with a mean of 8.3 more days of psychiatric hospitalization over the four-year period, representing 61 percent more hospital days. Medicaid beneficiaries who experience interruptions in coverage have, on average, a significantly greater use of inpatient psychiatric services while participating in Medicaid than beneficiaries with continuous Medicaid coverage. These findings suggest potential benefits of maintaining continuous Medicaid eligibility for beneficiaries with a severe mental illness.


Comparison of clinicopathologic features and survival of histopathologically amelanotic and pigmented melanomas: a population-based study.
IMPORTANCE Previous studies have reported that histopathologically amelanotic melanoma is associated with poorer survival than pigmented melanoma; however, small numbers of amelanotic melanomas, selected populations, lack of centralized pathologic review, or no adjustment for stage limit the interpretation or generalization of results from prior studies.OBJECTIVE To compare melanoma-specific survival between patients with histopathologically amelanotic and those with pigmented melanoma in a large international population-based study.DESIGN, SETTING, AND PARTICIPANTS Survival analysis with a median follow-up of 7.6 years.The study population comprised 2995 patients with 3486 invasive primary melanomas centrally scored for histologic pigmentation from the Genes, Environment, and Melanoma(GEM) Study, which enrolled incident cases of melanoma diagnosed in 1998 through 2003 from international population-based cancer registries.MAIN OUTCOMES AND MEASURES Clinicopathologic predictors and melanoma-specific survival of histologically amelanotic and pigmented melanoma were compared using generalized estimating equations and Cox regression models, respectively.RESULTS Of 3467 melanomas, 275 (8%) were histopathologically amelanotic. Female sex,nodular and unclassified or other histologic subtypes, increased Breslow thickness, presence of mitoses, severe solar elastosis, and lack of a coexisting nevus were independently associated with amelanotic melanoma (each P < .05). Amelanotic melanoma was generally ofa higher American Joint Committee on Cancer (AJCC) tumor stage at diagnosis (odds ratios[ORs] [95%CIs] between 2.9 [1.8-4.6] and 11.1 [5.8-21.2] for tumor stages between T1b and T3b and ORs [95%CIs] of 24.6 [13.6-44.4] for T4a and 29.1 [15.5-54.9] for T4b relative to T1a;P value for trend, <.001) than pigmented melanoma. Hazard of death from melanoma was higher for amelanotic than for pigmented melanoma (hazard ratio [HR], 2.0; 95%CI, 1.4-3.0)(P < .001), adjusted for age, sex, anatomic site, and study design variables, but survival did not differ once AJCC tumor stage was also taken into account (HR, 0.8; 95%CI, 0.5-1.2)(P = .36).CONCLUSIONS AND RELEVANCE At the population level, survival after diagnosis of amelanotic melanoma is poorer than after pigmented melanoma because of its more advanced stage at diagnosis. It is probable that amelanotic melanomas present at more advanced tumor stages because they are difficult to diagnose. The association of amelanotic melanoma with presence of mitoses independently of Breslow thickness and other clinicopathologic characteristics suggests that amelanotic melanomas might also grow faster than pigmented melanomas. New strategies for early diagnosis and investigation of the biological properties of amelanotic melanoma are warranted.


Strange Bedfellows: Nuclear Factor, Erythroid 2-Like 2 (Nrf2) and Hypoxia-Inducible Factor 1 (HIF-1) in Tumor Hypoxia.
The importance of the tumor microenvironment for cancer progression and therapeutic resistance is an emerging focus of cancer biology. Hypoxia, or low oxygen, is a hallmark of solid tumors that promotes metastasis and represents a significant obstacle to successful cancer therapy. In response to hypoxia, cancer cells activate a transcriptional program that allows them to survive and thrive in this harsh microenvironment. Hypoxia-inducible factor 1 (HIF-1) is considered the main effector of the cellular response to hypoxia, stimulating the transcription of genes involved in promoting angiogenesis and altering cellular metabolism. However, growing evidence suggests that the cellular response to hypoxia is much more complex, involving coordinated signaling through stress response pathways. One key signaling molecule that is activated in response to hypoxia is nuclear factor, erythroid 2 like-2 (Nrf2). Nrf2 is a transcription factor that controls the expression of antioxidant-response genes, allowing the cell to regulate reactive oxygen species. Nrf2 is also activated in various cancer types due to genetic and epigenetic alterations, and is associated with poor survival and resistance to therapy. Emerging evidence suggests that coordinated signaling through Nrf2 and HIF-1 is critical for tumor survival and progression. In this review, we discuss the distinct and overlapping roles of HIF-1 and Nrf2 in the cellular response to hypoxia, with a focus on how targeting Nrf2 could provide novel chemotherapeutic modalities for treating solid tumors.


Establishment of goat embryonic stem cells from in vivo produced blastocyst-stage embryos.
Embryonic stem (ES) cells with the capacity for germ line transmission have only been verified in mouse and rat. Methods for derivation, propagation, and differentiation of ES cells from domestic animals have not been fully established. Here, we describe derivation of ES cells from goat embryos. In vivo-derived embryos were cultured on goat fetal fibroblast feeders. Embryos either attached to the feeder layer or remained floating and expanded in culture. Embryos that attached showed a prominent inner cell mass (ICM) and those that remained floating formed structures resembling ICM disks surrounded by trophectodermal cells. ICM cells and embryonic disks were isolated mechanically, cultured on feeder cells in the presence of hLIF, and outgrown into ES-like colonies. Two cell lines were cultured for 25 passages and stained positive for alkaline phosphatase, POU5F1, NANOG, SOX2, SSEA-1, and SSEA-4. Embryoid bodies formed in suspension culture without hLIF. One cell line was cultured for 2 years (over 120 passages). This cell line differentiated in vitro into epithelia and neuronal cells, and could be stably transfected and selected for expression of a fluorescent marker. When cells were injected into SCID mice, teratomas were identified 5-6 weeks after transplantation. Expression of known ES cell markers, maintenance in vitro for 2 years in an undifferentiated state, differentiation in vitro, and formation of teratomas in immunodeficient mice provide evidence that the established cell line represents goat ES cells. This also is the first report of teratoma formation from large animal ES cells.


Predicting falls in community-dwelling older adults: a systematic review of prognostic models.
To systematically review and critically appraise prognostic models for falls in community-dwelling older adults. Prospective cohort studies with any follow-up period. Studies had to develop or validate multifactorial prognostic models for falls in community-dwelling older adults (60+ years). Models had to be applicable for screening in a general population setting. MEDLINE, EMBASE, CINAHL, The Cochrane Library, PsycINFO and Web of Science for studies published in English, Danish, Norwegian or Swedish until January 2020. Sources also included trial registries, clinical guidelines, reference lists of included papers, along with contacting clinical experts to locate published studies. Two authors performed all review stages independently. Data extraction followed the Critical Appraisal and Data Extraction for Systematic Reviews of Prediction Modelling Studies checklist. Risk of bias assessments on participants, predictors, outcomes and analysis methods followed Prediction study Risk Of Bias Assessment Tool. After screening 11 789 studies, 30 were eligible for inclusion (n=86 369 participants). Median age of participants ranged from 67.5 to 83.0 years. Falls incidences varied from 5.9% to 59%. Included studies reported 69 developed and three validated prediction models. Most frequent falls predictors were prior falls, age, sex, measures of gait, balance and strength, along with vision and disability. The area under the curve was available for 40 (55.6%) models, ranging from 0.49 to 0.87. Validated models' The area under the curve ranged from 0.62 to 0.69. All models had a high risk of bias, mostly due to limitations in statistical methods, outcome assessments and restrictive eligibility criteria. An abundance of prognostic models on falls risk have been developed, but with a wide range in discriminatory performance. All models exhibited a high risk of bias rendering them unreliable for prediction in clinical practice. Future prognostic prediction models should comply with recent recommendations such as Transparent Reporting of a multivariable prediction model for Individual Prognosis or Diagnosis. CRD42019124021.


Segmental distribution in refractory ulcerative colitis: a histological evaluation in pediatric and adult patients who underwent proctocolectomy.
Histological segmental distribution of ulcerative colitis (UC) may represent an early finding of newly diagnosed UC, especially in pediatric patients; a sign of Crohn's disease; or a predictor of refractory UC. However, its significance remains unknown. This study was performed to compare the final pathologic findings in resected total colon and rectum specimens between pediatric and adult patients with refractory UC who underwent proctocolectomy and clarify the significance of segmental distribution in pediatric patients with UC. Ninety patients with medically refractory UC (14 with childhood-onset UC and 76 with adult-onset UC) who underwent a two-staged operation comprising total proctocolectomy and ileal J-pouch anal anastomosis in the first stage were included in this study. We retrospectively reviewed all patients' medical records and performed a histological evaluation of the resected total colon using Geboes' grading system for histological inflammation activity of UC and factors of Tanaka's criteria for refractory UC. Histological segmental distribution was more frequently observed in the resected colon at surgery in pediatric (64%) than in adult patients (21%) with refractory UC. The administration of immunomodulators or anti-TNF-α antibody within 30 days before surgery was more frequently performed in adults than in children. Patients with refractory UC with a segmental distribution had a statistically significant relationship with shorter duration before surgery (P = 0.0009), male gender (P = 0.0342), and higher activity of disease inflammation in rectum (P = 0.0399). Multivariate analysis revealed the statistically significant relationship between segmental distribution and disease duration before surgery ≤ 6 years (P = 0.0031). Endoscopists should perform biopsies of both normal and abnormal mucosa throughout the total colon and follow changes in the pathological pattern because segmental distribution is an important predictor of prognosis.


Maternal mortality in the developed world: a review of surveillance methods, levels and causes of maternal deaths during 2006-2010.
This article provides an overview of surveillance methods, levels, and causes of maternal mortality in developed countries, in Europe and the United States, during 2006-2010. Accurate identification of maternal deaths is not always possible, and no two countries in the world use the same surveillance methods for maternal mortality. Despite limitations (e.g. underestimation, misclassification), routine vital registration systems are the backbone of maternal mortality surveillance systems in developed countries. Enhanced surveillance methods involve linkages between deaths of women of reproductive age and births within the preceding year, or the use of additional data sources for maternal deaths. Confidential enquiries into maternal deaths, in place in France, the Netherlands, and the UK are the gold standard in maternal mortality surveillance. Levels of maternal mortality in Europe were the lowest in the world during 2006-2010. While Europe has not seen major changes in maternal mortality in recent years, pregnancy-related mortality increased considerably in the USA, where improvements in the identification of deaths appear to play a part. The triad of infection, hemorrhage, and hypertensive disorders of pregnancy, which in the past accounted for >90% of all maternal deaths, now accounts for 60-70% of such deaths in developed countries. Maternal mortality surveillance provides learning opportunities to prevent future maternal deaths. There is need for integration of linked, multiple data sources into current maternal mortality surveillance systems to improve their utility.


Advances in Hair Restoration.
Hair loss affects millions of people worldwide and can have devastating effects on an individual's psychological and emotional well-being. Hair restoration technologies have advanced with the use of robotics and manual and motorized follicular unit extraction to provide patients excellent clinical results. Adjuvant modalities such as platelet-rich plasma injections, lasers and stem cells can further enhance the survivability and appearance of hair transplants.


Prevalence of and Risk Factors for Intimate Partner Violence in the First 6 Months Following HIV Diagnosis Among a Population-Based Sample in Rural Uganda.
Research in Uganda examining HIV-positive status disclosure and IPV victimization is scarce, and existing findings may not generalize to community-based samples of men and women newly diagnosed with HIV in Uganda. We investigated the prevalence of lifetime IPV, IPV experienced between HIV diagnosis and 6 months following diagnosis (recent IPV), and IPV specifically related to a partner learning one's HIV-positive status among a sample of men and women newly diagnosed with HIV in a population-based study in rural Uganda. We also examined correlates of recent IPV, including HIV-positive status disclosure. The sample included 337 participants followed for 6 months after HIV diagnosis. Lifetime IPV findings showed that over half of the sample reported experiencing emotional IPV (62.81% of men, 70.37% of women), followed by physical IPV (21.49% of men, 26.39% of women) then sexual IPV (7.44% of men, 17.59% of women). For recent IPV, men and women reported similar rates of physical (4.63% and 8.29%, respectively) and emotional (19.44% and 25.91%, respectively) IPV. Women were more likely than men to report recent sexual IPV (8.29% vs. 1.85%); however, this relationship was no longer significant after controlling for other risk factors associated with sexual IPV (AOR = 3.47, 95% CI [0.65, 18.42]). Participants who disclosed their HIV-positive status to their partner had 59% lower odds of reporting emotional IPV (AOR = 0.41, 95% CI [0.21, 0.81]) than participants who did not disclose their HIV-positive status. Younger age, non-polygamous marriage, lower social support, and greater acceptance for violence against women were also significantly associated with experience of recent IPV. Overall, 12.20% of participants who experienced recent IPV reported that the IPV was related to their partner learning their HIV-positive status. Findings highlight the need for IPV screening and intervention integrated into HIV diagnosis, care, and treatment services.


Biophysical characterization of recombinant HIV-1 subtype C virus infectivity factor.
HIV-1 virus infectivity factor (Vif) is one of the four accessory proteins that are characteristic of primate lentiviruses and critically required for the infection of host cells. Vif plays a key role in replication and transmission of the virus in non-permissive cells, such as primary T cells and macrophages. Using co-precipitation and co-fractionation techniques, evidence has been provided that Vif interacts with a variety of host proteins, such as the cytidine deaminases APOBEC3G and 3F, the Cullin5/EloBC ubiquitin-ligase complex, Fyn and Hck tyrosine kinases, as well as with viral components, such as the immature Gag precursor and viral RNA. We report on the expression, purification and molecular characterization of a folded recombinant subtype C Vif. Vif was expressed in E. coli with a C-terminal hexahistidine tag and purified by nickel affinity chromatography. We obtained approximately 5 mg protein per liter of bacterial culture, with a purity >95%. The expected molecular mass of 23.7 kDa was confirmed by mass spectrometry. Although dynamic light scattering and small angle X-ray scattering measurements revealed the presence of high molecular weight aggregates in the protein preparation, circular dichroism analysis showed that the protein contains mainly folded β-sheet elements and exhibits remarkable thermal stability (T (m) > 95°C). Recombinant Vif may be used as a tool to study its biological functions and tertiary structure, as well as for the development of diagnostic, therapeutic and preventive strategies for HIV-1 infections.


Concentrating colloids with electric field gradients. II. Phase transitions and crystal buckling of long-ranged repulsive charged spheres in an electric bottle.
We explored the usefulness of electric field gradients for the manipulation of the particle concentration in suspensions of charged colloids, which have long-ranged repulsive interactions. In particular, we studied the compression obtained by "negative" dielectrophoresis, which drives the particles to the regions of lowest field strength, thus preventing unwanted structural changes by induced dipole-dipole interactions. We used several sample cell layouts and suspension compositions, with a different range of the interparticle repulsions. For these systems, we obtained sufficient compression to observe a transition from the initial fluid phase to a random hexagonal close-packed crystal, as well as a body-centered cubic crystal. The heterogeneous dielectrophoretic crystallization mechanism involved an intriguing "pluglike" motion of the crystal, similar to what we have previously reported for hard-sphere suspensions. In this way, remarkably large single crystals were formed of several millimeters wide and a couple of centimeters long. Moreover, we found that these crystals could be compressed to such an extent that it led to an anisotropic deformation ("buckling") and, upon subsequent relaxation, a reorientation of the lattice, while stacking errors disappeared. These striking differences with the compressed hard-sphere crystals that we studied before [M. E. Leunissen et al., J. Chem. Phys. 128, 164508 (2008).] are likely due to the smaller elastic moduli of the present lower-density soft-sphere crystals.


The effect of pentoxifylline on cochlear blood flow.
Pentoxifylline, a phosphodiesterase inhibitor and hemorrheologic agent has been found to increase oxygen delivery to ischemic tissue. Intravenous pentoxifylline was administered to normal guinea pigs in order to assess the effect of pentoxifylline on cochlear blood flow and to elucidate its mechanism of action. Intravenous pentoxifylline was found to acutely increase cochlear blood flow in a dose-dependent manner. In normal animals, the effect appeared strongly related to the rheologic properties of this agent rather than a vasodilative action. Normovolemic hemodilution with 75% dextran resulted in no increase in cochlear blood flow during infusion of pentoxifylline, whereas the application of nitroprusside over the round window failed to abolish the effect of pentoxifylline.


Electrically controlled eight-spin-qubit entangled-state generation in a molecular break junction.
The generation of spin-based multi-qubit entangled states in the presence of an electric field is one of the most challenging tasks in current quantum-computing research. Such examples are still elusive. By using non-equilibrium Green's function-based quantum-transport calculations in combination with non-collinear spin density functional theory, we report that an eight-spin-qubit entangled state can be generated with the high-spin state of a dinuclear Fe(II) complex when the system is placed in a molecular break junction. The possible gate operation scheme, gating time, and decoherence issues have been carefully addressed. Furthermore, our calculations reveal that the preservation of the high spin state of this complex is possible if the experimentalists keep the electric-field strength below 0.78 V nm(-1). In brief, the present study offers a unique way to realize the first example of a multi-qubit entangled state by electrical means only.


The impact of patients as trainers on registered nurses' patient engagement in primary care clinics: a qualitative study.
In Canada, primary care is usually the front door to health care for people with health issues. Among these primary care services are primary care clinics (PCC), where the competencies of registered nurses (RNs) are needed. However, nursing practice in PCCs is variable and sometimes suboptimal from one PCC to another. In 2019, the Quebec Ministry of Health and Social Services deployed a practical guide for RNs practicing in PCCs. This guide was intended to support best professional and interprofessional practices and enhance the quality of services offered according to a physical-social vision of care, interprofessional collaboration and partnership with the patient. The Formation de formateurs en première ligne (F2PL) project team developed a train-the-trainer educational intervention to support RNs in assimilating the content of this guide. This educational intervention is uncommon because it includes patients as trainers (PTs). PTs developed and provided andragogic content about patient's experience to enhance patient engagement. To describe the impacts of the educational intervention provided by the PTs in nurses' patient engagement practices in PCCs. A descriptive qualitative approach was used to describe in-depth changes in RNs' practices. Individual interviews were conducted with 10 RNs and 3 PTs to explore the changes in RNs' practice and the barriers and facilitators to adopting this new practice. An inductive and deductive thematic analysis was carried out according to a conceptual model of patient engagement (the Montreal Model), and emerging themes were condensed into propositions. To ensure credibility, a peer review was conducted with the F2PL team, which includes a patient co-leader. The educational intervention provided by PTs has impacted RNs' practice in 3 ways: awareness or reminding of general principles, updating commitment to already known principles and enhancing the development of new professional skills. PTs could effectively support the RNs' motivation to use patient engagement practices in primary care.


Common drug interactions leading to adverse drug events in the intensive care unit: management and pharmacokinetic considerations.
Critically ill patients are predisposed to drug interactions because of the complexity of the drug regimens they receive in the intensive care setting. Drugs may affect the absorption, distribution, metabolism, and/or elimination of an object drug and consequently alter the intended pharmacologic response and potentially lead to an adverse event. Certain disease states that afflict critically ill patients may also amplify an intended pharmacologic response and potentially result in an unintended effect. A team approach is important to identify, prevent, and address drug interactions in the intensive care setting and optimize patient outcomes.


Risk assessment of airborne transmission of COVID-19 by asymptomatic individuals under different practical settings.
The lack of quantitative risk assessment of airborne transmission of COVID-19 under practical settings leads to large uncertainties and inconsistencies in our preventive measures. Combining in situ measurements and computational fluid dynamics simulations, we quantify the exhaled particles from normal respiratory behaviors and their transport under elevator, small classroom, and supermarket settings to evaluate the risk of inhaling potentially virus-containing particles. Our results show that the design of ventilation is critical for reducing the risk of particle encounters. Inappropriate design can significantly limit the efficiency of particle removal, create local hot spots with orders of magnitude higher risks, and enhance particle deposition causing surface contamination. Additionally, our measurements reveal the presence of a substantial fraction of faceted particles from normal breathing and its strong correlation with breathing depth.


Effect of homogenisation in formation of thermally induced aggregates in a non- and low- fat milk model system with microparticulated whey proteins.
The objective of the research presented in this paper was to investigate how different characteristics of whey protein microparticles (MWP) added to milk as fat replacers influence intermolecular interactions occurring with other milk proteins during homogenisation and heating. These interactions are responsible for the formation of heat-induced aggregates that influence the texture and sensory characteristics of the final product. The formation of heat-induced complexes was studied in non- and low-fat milk model systems, where microparticulated whey protein (MWP) was used as fat replacer. Five MWP types with different particle characteristics were utilised and three heat treatments used: 85 °C for 15 min, 90 °C for 5 min and 95 °C for 2 min. Surface characteristics of the protein aggregates were expressed as the number of available thiol groups and the surface net charge. Intermolecular interactions involved in the formation of protein aggregates were studied by polyacrylamide gel electrophoresis and the final complexes visualised by darkfield microscopy. Homogenisation of non-fat milk systems led to partial adsorption of caseins onto microparticles, independently of the type of microparticle. On the contrary, homogenisation of low-fat milk resulted in preferential adsorption of caseins onto fat globules, rather than onto microparticles. Further heating of the milk, led to the formation of heat induced complexes with different sizes and characteristics depending on the type of MWP and the presence or not of fat. The results highlight the importance of controlling homogenisation and heat processing in yoghurt manufacture in order to induce desired changes in the surface reactivity of the microparticles and thereby promote effective protein interactions.


Conformal and stereotactic radiotherapy in hepatocellular carcinoma.
Historically, Radiation Therapy (RT) has played a minor role in the treatment of hepatocellular carcinoma, especially because of toxicity related to conventional techniques. However, high radiation doses can now be given safely by means of 3D-conformal techniques and other more specialized techniques. 3D-conformal RT, in which both target definition and treatment planning are based on three-dimensional image data sets, provided good local control in several series. As an evolution of cranial radiosurgery, stereotactic body radiation therapy (SBRT) is currently being employed as a new tool for the treatment of HCC, as it offers the opportunity of achieving radioablation for tumors up to 6 cm in diameter with a minimal risk of radiation induced liver disease. SBRT is distinguished by the fact that both planning and treatment are guided by reference to a system of external stereotactic coordinates. As a consequence, SBRT may succeed in minimizing geometric treatment uncertainties, thus allowing the delivery of high doses to the target, given in a small number of fractions. In several published series of SBRT for HCC, actuarial 5-year local control rates higher than 80% have been achieved In short, radiation therapy should be considered as a useful addition to the treatment spectrum of HCC.


A novel allosteric inhibitor that prevents IKKβ activation.
I kappa B kinase β (IKKβ) is one of the primary targets to regulate canonical NF-κB activity. The misregulation of NF-κB is associated with various diseases, including chronic inflammation and cancers. Most of the known IKKβ inhibitors target its active form and suffer from poor selectivity. In the present study, we aim to design inhibitors that can bind to the IKKβ inactive form and block its activation. We identified a potential allosteric site between the kinase domain (KD) and ubiquitin-like domain (ULD) of human IKKβ and used it to virtually screen a chemical library for allosteric inhibitors. Among the 133 compounds tested, 16 inhibited NF-κB activity by over 50% at 50 μM in a reporter gene assay. Further quantitative measurements and cytotoxicity study gave one compound 124 (3,4-dichloro-2-ethoxy-N-(2,2,6,6-tetramethylpiperidin-4-yl)benzenesulfonamide) which specifically targets the IKKβ inactive form. In cells, 124 inhibited IκBα phosphorylation and NF-κB transcriptional activity for the reporter gene with an IC50 of 35 μM by decreasing the phosphorylation level of Ser177/181 on IKKβ and blocking its activation upon TNFα stimulation. Molecular dynamics simulations demonstrated that 124 binds to the pocket between KD and ULD in the inactive conformation of IKKβ rather than the active conformation. As the first allosteric inhibitor that prevents IKKβ activation, 124 provides a good starting point for further inhibitor discovery and a probe for IKKβ enzyme cycle and regulatory mechanism study.


Botulinum toxin for the treatment of gastroparesis: a preliminary report.
Gastroparesis is a disorder of gastric motility that results in delayed gastric emptying. Common symptoms include early satiety, postprandial fullness, epigastric pain, nausea, vomiting, and weight loss. The underlying etiologies of gastroparesis are many and include diabetes, prior gastric surgery, collagen vascular disorders, and a previous viral illness. Up to one third of cases are classified as idiopathic. Treatment typically consists of a change in diet to small volume, frequent meals and the use of the prokinetic agents metoclopramide, cisapride, erythromycin, or domperidone. Botulinum toxin has recently been shown to be effective in treating disorders of smooth muscle hypertonicity in the GI tract. This case report describes three patients with severe gastroparesis whose symptoms persisted despite dietary changes and the use of high dose prokinetic agents. All three were treated with intrasphincteric injection of the pylorus with botulinum toxin and all had significant symptomatic improvement afterwards. Possible mechanisms of action of botulinum toxin on the pylorus and its effects in patients with gastroparesis are discussed.


GPR40 (FFAR1) - Combined Gs and Gq signaling in vitro is associated with robust incretin secretagogue action ex vivo and in vivo.
GPR40 (FFAR1), a clinically proven anti-diabetes target, is a Gq-coupled receptor for long chain fatty acids (LCFA) stimulating insulin secretion directly and mediating a major part of the dietary triglyceride-induced secretion of the incretins GLP-1 and GIP. In phase-II studies the GPR40 agonist TAK-875 decreased blood glucose but surprisingly without stimulating incretins. Here we find that GPR40 can signal through not only Gq and IP3 but also Gs and cAMP when stimulated with certain agonists such as AM-1638 and AM-5262 in contrast to the endogenous LCFA ligands and agonists such as TAK-875 and AM-837, which only signal through Gq. In competition binding against [3H]AM-1638 and [3H]L358 the Gq + Gs and the Gq-only agonists either competed for or showed positive cooperativity by increasing the binding of the two different radio-ligands, in opposite ways. Nevertheless, both the Gq-only and the Gq + Gs agonists all docked surprisingly well into the binding site for TAK-875 in the X-ray structure of GPR40. In murine intestinal primary cell-cultures the endogenous LCFAs and the Gq-only agonists stimulated GLP-1 secretion with rather poor efficacy as compared with the high efficacy Gq + Gs GPR40 agonists and a prototype GPR119 agonist. Similarly, in fasting both male and female mice the Gq + Gs agonists showed significantly higher efficacy than the Gq-only agonists in respect of increasing plasma GLP-1 and plasma GIP in a GPR40-dependent manner. It is concluded that stimulation of GPR40 by endogenous LCFAs or by Gq-only synthetic agonists result in a rather limited incretin response, whereas Gq + Gs GPR40 agonists stimulate incretin secretion robustly.


High-Level Teleoperation System for Aerial Exploration of Indoor Environments.
Exploration of challenging indoor environments is a demanding task. While automation with aerial robots seems a promising solution, fully autonomous systems still struggle with high-level cognitive tasks and intuitive decision making. To facilitate automation, we introduce a novel teleoperation system with an aerial telerobot that is capable of handling all demanding low-level tasks. Motivated by the typical structure of indoor environments, the system creates an interactive scene topology in real-time that reduces scene details and supports affordances. Thus, difficult high-level tasks can be effectively supervised by a human operator. To elaborate on the effectiveness of our system during a real-world exploration mission, we conducted a user study. Despite being limited by real-world constraints, results indicate that our system better supports operators with indoor exploration, compared to a baseline system with traditional joystick control.


Neuronal Encoding of Speech Features in the Human Thalamus in Parkinson's Disease and Essential Tremor Patients.
The human thalamus is known, from stimulation studies and functional imaging, to participate in high-level language tasks. The goal of this study is to find whether and how speech features, in particular, vowel phonemes, are encoded in the neuronal activity of the thalamus, and specifically of the left ventralis intermediate nucleus (Vim), during speech production, perception, and imagery. In this cross-sectional study, we intraoperatively recorded single neuron activity in the left Vim of eight neurosurgical patients with Parkinson's disease (PD) (n = 4) or essential tremor (n = 4) undergoing implantation of deep brain stimulation (n = 3) or radiofrequency lesioning (n = 5) while patients articulated the five monophthongal vowel sounds. In this article, we report that single neurons in the left Vim encode individual vowel phonemes mainly during speech production but also during perception and imagery. They mainly use one of two encoding schemes: broad or sharp tuning, with a similar percentage of units each. Sinusoidal tuning has been demonstrated in almost half of the broadly tuned units. Patients with PD had a lower percentage of speech-related units in each aspect of speech (production, perception, and imagery), a significantly lower percentage of broadly tuned units, and significantly lower median firing rates during speech production and perception, but significantly higher rates during imagery, than patients with essential tremor. The results suggest that the left Vim uses mixed encoding schemes for speech features. Our findings explain, at the single neuron level, why deep brain stimulation and radiofrequency lesioning of the left Vim are likely to cause speech side effects. Moreover, they may indicate that speech-related units in the left Vim of patients with PD may be degraded even in the subclinical phase.


Extracellular high mobility group box-1 inhibits R5 and X4 HIV-1 strains replication in mononuclear phagocytes without induction of chemokines and cytokines.
High mobility group box-1 (HMGB1) is a nuclear chromatin protein. Furthermore, it induces chemotaxis and inflammation once released in the extracellular milieu, and it has been reported to upregulate, but also to inhibit HIV-1 replication in different cell types. We here investigated the potential role of extracellular HMGB1 in both R5 and X4 HIV-1 replication in primary human monocyte-derived macrophages (MDM) and U937 promonocytic cells, respectively. MDM or U937 cells were infected with R5 and X4 HIV-1 strains, respectively, in the presence or absence of endotoxin-free recombinant (r) HMGB1 or necrotic cell supernatants either containing or depleted of endogenous HMGB1. HIV replication was measured by means of virion-associated reverse transcriptase activity in culture supernatants and cell-associated viral protein expression. Cytokine and chemokine production were measured by enzyme-linked immunosorbent assay; cell surface expression of CD4, CC chemokine receptor 5, receptor for advanced glycation end-products, Toll-like receptor-2 and Toll-like receptor-4 were analyzed by flow cytometry. Both rHMGB1 and necrotic cell supernatant-associated HMGB1 inhibited replication of R5 HIV-1 in MDM. Surprisingly enough, no upregulation of CC chemokine receptor 5-binding chemokines or of other chemokines and cytokines was observed in rHMGB1-stimulated MDM. HMGB1 also induced chemotaxis and strongly inhibited the replication of X4 HIV-1 in the 'Minus' subset of U937 cell clones expressing high levels of putative HMGB1 receptors (receptor for advanced glycation end-products, Toll-like receptors 2 and 4). Extracellular HMGB1 is a potent inhibitor of both R5 and X4 HIV-1 replication in mononuclear phagocytic cells without inducing the release of HIV-Modulatory chemokines or cytokines.


Antimicrobial peptides in reptiles.
Reptiles are among the oldest known amniotes and are highly diverse in their morphology and ecological niches. These animals have an evolutionarily ancient innate-immune system that is of great interest to scientists trying to identify new and useful antimicrobial peptides. Significant work in the last decade in the fields of biochemistry, proteomics and genomics has begun to reveal the complexity of reptilian antimicrobial peptides. Here, the current knowledge about antimicrobial peptides in reptiles is reviewed, with specific examples in each of the four orders: Testudines (turtles and tortosises), Sphenodontia (tuataras), Squamata (snakes and lizards), and Crocodilia (crocodilans). Examples are presented of the major classes of antimicrobial peptides expressed by reptiles including defensins, cathelicidins, liver-expressed peptides (hepcidin and LEAP-2), lysozyme, crotamine, and others. Some of these peptides have been identified and tested for their antibacterial or antiviral activity; others are only predicted as possible genes from genomic sequencing. Bioinformatic analysis of the reptile genomes is presented, revealing many predicted candidate antimicrobial peptides genes across this diverse class. The study of how these ancient creatures use antimicrobial peptides within their innate immune systems may reveal new understandings of our mammalian innate immune system and may also provide new and powerful antimicrobial peptides as scaffolds for potential therapeutic development.


Synthesis and characterization of a matrix-metalloproteinase responsive silk-elastinlike protein polymer.
Silk-elastinlike protein polymers (SELPs) are recombinant polymers consisting of tandem repeats of silk (GAGAGS) and elastin (GVGVP) units. By modification of the length and composition of these repeats, the properties of SELP hydrogels can be controlled for specific applications including nucleic acid and virus delivery and tissue engineering. Here, the structure of SELPs is further modified to include a sequence that is sensitive to matrix-metalloproteinases (MMPs). MMPs are a ubiquitous family of extracellular matrix-modifying enzymes that are commonly associated with numerous vital processes. Increased levels of MMPs are found at high levels locally in many types of solid tumors. By modifying the SELP backbone with MMP-sensitive peptide sequences, a hydrogel that is degradable by MMPs was produced. The MMP-sensitivity of the polymer was examined by incubation with MMP-2 and MMP-9, which yielded complete cleavage of all full-length polymers by 36 hours and 48 hours, respectively, with no observable effect on unmodified SELP. Hydrogel sensitivity was tested by exposure to MMP-2 or MMP-9 for 2 weeks, during which samples were taken to analyze protein loss from the hydrogel and release of 100 nm fluorescent beads. Following the incubation period, hydrogels were tested in mechanical compression to examine the loss of hydrogel stiffness due to degradation. It was found that MMP-2 and MMP-9 caused 63% and 44% increased protein loss and 65% and 95% increased release from MMP-sensitive hydrogels, while the compressive modulus decreased by 41% and 29%. These results suggest the potential of MMP-responsive SELPs for localized delivery of bioactive agents where MMPs are overexpressed.


Occlusion-robust Sleep Posture Detection using Body Rolling Motion in a Video.
It has been reported that the monitoring of sleep postures is useful for the treatment and prevention of sleep diseases such as obstructive sleep apnea and heart failure. Camera-based sleep posture detection is attractive for the nature of comfort and convenience of use. However, the main challenge is to detect postures from images of the body that are occluded by bed sheets or covers. To address this issue, we propose a novel occlusion-robust sleep posture detection method exploiting the body rolling motion in a video. It uses the head orientation to indicate the posture direction (supine, left or right lateral), triggered by the full-body rolling motion (as a sign of posture change). The experimental results show that our proposed method, as compared with the state-of-the-art approaches such as skeleton-based (MediaPipe) and full-image ResNet based methods, obtained clear improvements on sleep posture detection with heavy body occlusions, with an averaged precision, recall and F1-score of 0.974, 0.993 and 0.983, respectively. The next step is to integrate the sleep posture detection algorithm into a camera-based sleep monitoring system for clinical validations.


Leader sequence distinguishes between translatable and encapsidated measles virus RNAs.
The 3'-terminal 55 nucleotides of the negative-strand measles virus RNA genome called the leader sequence is not transcribed into a detectable distinct RNA product. Most of the monocistronic N and bicistronic N-P RNAs lack the leader sequence. However, a subpopulation of the N and N-P RNAs and all of the antigenomes possess this leader. Here, we show that leader-containing subgenomic RNAs are functionally distinct from their leaderless counterparts. In measles virus-infected cells, leaderless monocistronic N and bicistronic N-P RNAs were associated with polysomes. By contrast, leader-containing N and N-P RNAs were found exclusively in nonpolysomal ribonucleoprotein complexes that were resistant to RNase and had a buoyant density of 1.30 g/ml, the same as that of antigenomic ribonucleoprotein complexes. Both antigenomic and subgenomic ribonucleoprotein complexes were specifically immunoprecipitated by antiserum against the N protein, and leaderless RNAs were not found in these complexes. These findings suggest that measles virus distinguishes RNAs destined for encapsidation or translation by the presence or absence of a leader sequence.


Twenty-first century ethical challenges for psychology.
Foreseeable social and technological changes will force us to reevaluate our thinking about ethically appropriate ways to fulfill our mission of using psychology to advance human health and welfare in the twenty-first century. Three categories of challenge related to societal and technological changes have become particularly evident. First, increasing patterns of delivering services over substantial distances by electronic means (i.e., telepsychology) demand consideration. Second, we must parse our ethical obligations to individuals, to groups, and to society at large as our influence working behind the scenes as "invisible" psychologists grows. Finally, as we witness the accelerating demise of psychiatry, we must take care not to follow a similar path. As we face new ethical challenges, we must continually ask ourselves where our responsibilities lie as individuals and as a profession. We must learn not to repeat the mistakes of the past and focus instead on optimizing the future for a science and practice of psychology focused on human health and welfare.


Genomic properties of chromosomal bands are linked to evolutionary rearrangements and new centromere formation in primates.
Chromosomal rearrangements in humans are largely related to pathological conditions, and phenotypic effects are also linked to alterations in the expression profile following nuclear relocation of genes between functionally different compartments, generally occupying the periphery or the inner part of the cell nuclei. On the other hand, during evolution, chromosomal rearrangements may occur apparently without damaging phenotypic effects and are visible in currently phylogenetically related species. To increase our insight into chromosomal reorganisation in the cell nucleus, we analysed 18 chromosomal regions endowed with different genomic properties in cell lines derived from eight primate species covering the entire evolutionary tree. We show that homologous loci, in spite of their evolutionary relocation along the chromosomes, generally remain localised to the same functional compartment of the cell nuclei. We conclude that evolutionarily successful chromosomal rearrangements are those that leave the nuclear position of the regions involved unchanged. On the contrary, in pathological situations, the effect typically observed is on gene structure alteration or gene nuclear reposition. Moreover, our data indicate that new centromere formation could potentially occur everywhere in the chromosomes, but only those emerging in very GC-poor/gene-poor regions, generally located in the nuclear periphery, have a high probability of being retained through evolution. This suggests that, in the cell nucleus of related species, evolutionary chromosomal reshufflings or new centromere formation does not alter the functionality of the regions involved or the interactions between different loci, thus preserving the expression pattern of orthologous genes.


Hypogastric artery ligation: a new perspective.
This study was conducted at the Lokmanya Tilak Municipal Medical College and General Hospital, Sion, Bombay. An analysis was made of the hypogastric artery ligations done at our institution from January 1985 to December 1990. A prospective pilot study beginning June 1989 was added to the existing data. The hemostasis achieved intraoperatively and the collateral circulation maintained after main trunk ligation vs selective anterior division ligation have been studied prospectively with pelvic angiography postoperatively. Pelvic angiography using the standard Seldinger technique was done 14 days posthypogastric artery ligation in emergency obstetric cases. In selected gynecologic cases posted for Wertheim's hysterectomy, pelvic angiography was done both before and after hypogastric artery ligation. All the collected data, including operative time, blood loss, and complications, have been tabulated and compared with other studies in the literature.


Antibody-drug conjugates for previously treated aggressive lymphomas: focus on polatuzumab vedotin.
Antibody-drug conjugates (ADCs) are immunoconjugates and comprise a monoclonal antibody that is chemically attached to a cytotoxic drug (or payload) via a stable chemical linker. Since the approval of the first ADC in 2000, there are now nine different approved agents and over 100 ADCs in the drug-development pipeline. This review briefly describes the ADCs approved for treatment of lymphoma and their distinguishing factors in terms of target, linker and payload. The clinical implications of the use of ADCs are also considered. Here, we focus on polatuzumab vedotin, an ADC targeted to CD79b, which is approved for the treatment of patients with relapsed/refractory diffuse large B-cell lymphoma (R/R DLBCL) who have received at least one (EU approval) or two (US approval) prior therapies and are not eligible for bone marrow transplantation. The characteristics of polatuzumab vedotin are discussed and clinical data are presented. The future of polatuzumab vedotin clinical development, and ADCs in general, are also considered. ADCs represent a significant advance in the treatment of lymphoma. Polatuzumab vedotin has shown clinical efficacy and a tolerable safety profile in both first-line and R/R DLBCL; future studies are planned to further investigate this ADC.


The Prevalence of Fibromyalgia Among Medical Students at King Abdulaziz University: A Cross-Sectional Study.
Background and objective Fibromyalgia (FM) is a chronic, multifactorial pain condition. The latest literature suggests that genetic and environmental factors including continuous stress contribute significantly to FM's pathophysiology. In this study, we aimed to investigate the prevalence of FM among medical students as they are considered a population significantly at risk of developing the condition. Methods This cross-sectional study was conducted at King Abdulaziz University. Medical students included in the study were recruited through a random stratified sampling method. A self-administered questionnaire was distributed to the participants; it included questions related to widespread pain index (WPI) and symptom severity scale (SSS) to assess the symptoms and diagnosis of FM, which were established based on the current diagnostic criteria. All first-year students were excluded from this research. Results A total of 450 participants were recruited for the study. Among them, 291 (64.7%) were females and 159 (35.3%) were males. Their ages ranged from 18 to 26 years, and the mean age was 21.52 years (SD: ±1.52). They came from different academic levels: 97 (21.6%) were in the second year, 79 (17.6%) were in the third year, 70 (15.6%) were in the fourth year, 99 (22%) were in the fifth year, and 105 (23.3%) were in the sixth year. The overall prevalence of FM was found to be 43 (9.6%). It was established based on the number of students who fulfilled the diagnostic criteria or were previously diagnosed with FM by a professional physician. Conclusion FM is highly prevalent among medical students. Our findings demonstrate the likelihood of the influence of medical school on causing the condition, as it has a stressful education system with high academic expectations. We recommend that this issue be seriously addressed since FM leads to a significant burden on the students and can negatively affect their future medical practice.


Monodisperse stimuli-responsive colloidosomes by self-assembly of microgels in droplets.
We introduce a novel and versatile technique to fabricate monodisperse stimuli-responsive colloidosomes using stimuli-responsive microgel particles as building blocks, aqueous droplets as templates, and microfluidic devices to control the assembly. Our colloidosomes exhibit approximately 80% decrease in volume when actuated; thus, they can be of immense potential in applications that require targeted pulsed-release of active materials. The use of microfluidics allows fabrication of extremely monodisperse colloidosomes. Alternatively, our technique can also be combined with bulk emulsification techniques to produce large quantities of colloidosomes for various applications.


Increased Risk of Malaria During the First Year of Life in Small-for-Gestational-Age Infants: A Longitudinal Study in Benin.
According to the Developmental Origins of Health and Diseases paradigm, the fetal period is highly vulnerable and may have profound effects on later health. Few studies assessed the effect of small-for-gestational age (SGA), a proxy for fetal growth impairment, on risk of malaria during infancy in Africa. We used data from a cohort of 398 mother-child pairs, followed from early pregnancy to age 1 year in Benin. Malaria was actively and passively screened using thick blood smear. We assessed the effect of SGA on risk of malaria infection and clinical malaria from birth to 12 months, after stratifying on the infant's age using a logistic mixed regression model. After adjustment for potential confounding factors and infant's exposure to mosquitoes, SGA was associated with a 2-times higher risk of malaria infection (adjusted odds ratio [aOR] = 2.16; 95% confidence interval [CI], 1.04-4.51; P = .039) and clinical malaria (aOR = 2.33; 95% CI, 1.09-4.98; P = .030) after age 6 months. Results suggest higher risk of malaria during the second semester of life in SGA infants, and argue for better follow-up of these infants after birth, as currently for preterm babies.


Initial Validation of a Modified MRI Scoring System for Assessing Outcomes after Single-Surface Osteochondral Shell Allograft Transplantation in the Knee.
The Osteochondral Allograft Magnetic Resonance Imaging Scoring System (OCAMRISS) provides a reproducible method for imaging-based grading for osteochondral allograft (OCA) transplants. However, the OCAMRISS does not account for larger whole-surface OCA shell grafts, and has not been validated for assessing outcomes after shell OCA transplantation. Therefore, the objective of this study was to evaluate a modified OCAMRISS for assessing single-surface shell OCAs in the knee based on strength of correlations for a modified OCAMRISS score with graft success and patient-reported outcomes for pain and function. With institutional review board approval and informed patient consent, patients who underwent large single-surface shell OCA transplantation and magnetic resonance imaging (MRI) exams at 1-year postsurgery were identified from a prospectively enrolled registry. All patients with a minimum of 2 years of clinical follow-up were included in the present study. A modified OCAMRISS, as well as assessment of the percentage of OCA bone incorporation, was used to score each knee. Two radiologists, blinded to patient demographics and outcomes, reviewed all MRIs together to determine a consensus score for each category and %-incorporation for each OCA. Thirteen patients (7 F, 6 M; mean age = 29.8 ± 9.4; mean body mass index = 27.1 ± 5.8); 8 medial femoral condyle, 4 patella, and 1 medial tibial plateau shell OCAs were evaluated. Mean modified OCAMRISS score was 5.2 ± 2.8, range (2-12) and %-integration was 72.7 ± 33.8, range (0-100). Moderate to strong correlations were noted for 1-year modified OCAMRISS total score with final follow-up (FFU) visual analog scale (VAS) pain (r = +0.58) and Single Assessment Numeric Evaluation (SANE) function (r = -0.7) scores, and for 1-year %-incorporation with FFU VAS pain (r = -0.76) and SANE function (r = +0.83) scores. The modified OCAMRISS total score and %-incorporation assessments determined at 1 year following single-surface shell OCA transplantation correlate well with initial patient outcomes and have clinical applicability for monitoring patients after large-shell OCA transplants in the knee.


Frequency and directionality of gene conversion events involving the CYC7-H3 mutation in Saccharomyces cerevisiae.
The CYC7-H3 mutation is a 5-kb deletion that causes overproduction of iso-2 cytochrome c. Unlike most mutations in yeast, the CYC7-H3 mutation is preferentially lost when it is involved in a gene conversion event. We have shown that cloned copies of CYC7-H3 DNA that are inserted into the yeast genome are associated with a high frequency of recombination and aberrant segregation events. Since parity in conversion frequency was observed when the extensive insertion/deletion heterozygosity at this locus was eliminated, we conclude that the CYC7-H3 sequences are inherently capable of acting as donors or recipients in gene conversion events, although they are unlikely to act as donors when they are located opposite a large heterology. DNA sequence comparisons revealed similarities between the CYC7-H3 junction region and the 2-micron circle DNA region that is involved in site-specific recombination.


Specific phobias in older adults: characteristics and differential diagnosis.
Differential diagnosis implies identifying shared and divergent characteristics between clinical states. Clinical work with older adults demands not only the knowledge of nosological features associated with differential diagnosis, but also recognition of idiosyncratic factors associated with this population. Several factors can interfere with an accurate diagnosis of specific phobia in older cohorts. The goal of this paper is to review criteria for specific phobia and its differential diagnosis with panic disorder, agoraphobia, post-traumatic stress disorder and obsessive compulsive disorder, while stressing the specific factors associated with aging. A literature search regarding specific phobia in older adults was carried out using PubMed. Relevant articles were selected and scanned for further pertinent references. In addition, relevant references related to differential diagnosis and assessment were used. Etiologic factors, specificity of feared stimulus or situation, fear predictability and the nature of phobic situations are key points to be assessed when implementing a differential diagnosis of specific phobia. First, age-related sensory impairments are common and interfere both with information processing and communication. Second, medical illnesses create symptoms that might cause, interfere with, or mimic anxiety. Third, cohort effects might result in underreporting, through the inability to communicate or recognize anxiety symptoms, misattributing them to physical conditions. Finally, diagnostic criteria and screening instruments were usually developed using younger samples and are therefore not adapted to the functional and behavioral characteristics of older samples.


Influence of microarrays experiments missing values on the stability of gene groups by hierarchical clustering.
Microarray technologies produced large amount of data. The hierarchical clustering is commonly used to identify clusters of co-expressed genes. However, microarray datasets often contain missing values (MVs) representing a major drawback for the use of the clustering methods. Usually the MVs are not treated, or replaced by zero or estimated by the k-Nearest Neighbor (kNN) approach. The topic of the paper is to study the stability of gene clusters, defined by various hierarchical clustering algorithms, of microarrays experiments including or not MVs. In this study, we show that the MVs have important effects on the stability of the gene clusters. Moreover, the magnitude of the gene misallocations is depending on the aggregation algorithm. The most appropriate aggregation methods (e.g. complete-linkage and Ward) are highly sensitive to MVs, and surprisingly, for a very tiny proportion of MVs (e.g. 1%). In most of the case, the MVs must be replaced by expected values. The MVs replacement by the kNN approach clearly improves the identification of co-expressed gene clusters. Nevertheless, we observe that kNN approach is less suitable for the extreme values of gene expression. The presence of MVs (even at a low rate) is a major factor of gene cluster instability. In addition, the impact depends on the hierarchical clustering algorithm used. Some methods should be used carefully. Nevertheless, the kNN approach constitutes one efficient method for restoring the missing expression gene values, with a low error level. Our study highlights the need of statistical treatments in microarray data to avoid misinterpretation.


HIV-1 Populations in Semen Arise through Multiple Mechanisms.
HIV-1 is present in anatomical compartments and bodily fluids. Most transmissions occur through sexual acts, making virus in semen the proximal source in male donors. We find three distinct relationships in comparing viral RNA populations between blood and semen in men with chronic HIV-1 infection, and we propose that the viral populations in semen arise by multiple mechanisms including: direct import of virus, oligoclonal amplification within the seminal tract, or compartmentalization. In addition, we find significant enrichment of six out of nineteen cytokines and chemokines in semen of both HIV-infected and uninfected men, and another seven further enriched in infected individuals. The enrichment of cytokines involved in innate immunity in the seminal tract, complemented with chemokines in infected men, creates an environment conducive to T cell activation and viral replication. These studies define different relationships between virus in blood and semen that can significantly alter the composition of the viral population at the source that is most proximal to the transmitted virus.


Estimation of SARS-CoV-2 Infection Fatality Rate by Real-time Antibody Screening of Blood Donors.
The pandemic due to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has tremendous consequences for our societies. Knowledge of the seroprevalence of SARS-CoV-2 is needed to accurately monitor the spread of the epidemic and to calculate the infection fatality rate (IFR). These measures may help the authorities make informed decisions and adjust the current societal interventions. The objective was to perform nationwide real-time seroprevalence surveying among blood donors as a tool to estimate previous SARS-CoV-2 infections and the population-based IFR. Danish blood donors aged 17-69 years giving blood 6 April to 3 May were tested for SARS-CoV-2 immunoglobulin M and G antibodies using a commercial lateral flow test. Antibody status was compared between geographical areas, and an estimate of the IFR was calculated. Seroprevalence was adjusted for assay sensitivity and specificity taking the uncertainties of the test validation into account when reporting the 95% confidence intervals (CIs). The first 20 640 blood donors were tested, and a combined adjusted seroprevalence of 1.9% (95% CI, .8-2.3) was calculated. The seroprevalence differed across areas. Using available data on fatalities and population numbers, a combined IFR in patients <70 years is estimated at 89 per 100 000 (95% CI, 72-211) infections. The IFR was estimated to be slightly lower than previously reported from other countries not using seroprevalence data. The IFR is likely severalfold lower than the current estimate. We have initiated real-time nationwide anti-SARS-CoV-2 seroprevalence surveying of blood donations as a tool in monitoring the epidemic.