Molecular Characterization of Notch1 Positive Progenitor Cells in the Developing Retina.
The oscillatory expression of Notch signaling in neural progenitors suggests that both repressors and activators of neural fate specification are expressed in the same progenitors. Since Notch1 regulates photoreceptor differentiation and contributes (together with Notch3) to ganglion cell fate specification, we hypothesized that genes encoding photoreceptor and ganglion cell fate activators would be highly expressed in Notch1 receptor-bearing (Notch1+) progenitors, directing these cells to differentiate into photoreceptors or into ganglion cells when Notch1 activity is diminished. To identify these genes, we used microarray analysis to study expression profiles of whole retinas and isolated from them Notch1+ cells at embryonic day 14 (E14) and postnatal day 0 (P0). To isolate Notch1+ cells, we utilized immunomagnetic cell separation. We also used Notch3 knockout (Notch3KO) animals to evaluate the contribution of Notch3 signaling in ganglion cell differentiation. Hierarchical clustering of 6,301 differentially expressed genes showed that Notch1+ cells grouped near the same developmental stage retina cluster. At E14, we found higher expression of repressors (Notch1, Hes5) and activators (Dll3, Atoh7, Otx2) of neuronal differentiation in Notch1+ cells compared to whole retinal cell populations. At P0, Notch1, Hes5, and Dll1 expression was significantly higher in Notch1+ cells than in whole retinas. Otx2 expression was more than thirty times higher than Atoh7 expression in Notch1+ cells at P0. We also observed that retinas of wild type animals had only 14% (P < 0.05) more ganglion cells compared to Notch3KO mice. Since this number is relatively small and Notch1 has been shown to contribute to ganglion cell fate specification, we suggested that Notch1 signaling may play a more significant role in RGC development than the Notch3 signaling cascade. Finally, our findings suggest that Notch1+ progenitors--since they heavily express both pro-ganglion cell (Atoh7) and pro-photoreceptor cell (Otx2) activators--can differentiate into either ganglion cells or photoreceptors.


Paclitaxel in anthracycline-treated breast cancer patients.
Between May 1995 and July 1997, paclitaxel (TX) (175 mg/m2 by 3 h i. v. infusion every 21 days) was administered to 70 consecutive patients (median age: 57 years) previously treated with the FEC regimen (cyclophosphamide and 5-fluorouracil, 600 mg/m2, plus epirubicin, 60 or 120 mg/m2) as an adjuvant setting or as a first-line therapy for metastatic disease. Sixty-eight patients were evaluable for response, while two died early. Patients received a median of 4.7 (3-12 course) of TX for a total of 211 courses. The overall response and stable disease rate was 54% in 11 patients, who relapsed following adjuvant FEC, and 60% in 57 patients, who received FEC as first treatment for their metastatic disease. No complete respose was obtained. In patients pretreated for metastatic disease, response and stable disease rates were similar irrespective of previous response to FEC. Main hematologic toxicity of TX was of short duration, grade II/III leukopenia (86% of patients) and non-hematologic toxicity was grade II/III peripheral neuropathy, related to the cumulative dose of TX. At this schedule, TX offers a significant rate of partial responses or disease stabilization in patients with metastatic breast cancer previously treated with FEC.


The effect of sleep duration on the risk of unintentional injury in Korean adults.
The decrease or increase in sleep duration has recently been recognized as a risk factor for several diseases, including hypertension and obesity. Many studies have explored the relationship of decreased sleep durations and injuries, but few have examined the relationship between increased sleep duration and injury. The objective of this research is to identify the risk for injury associated with both decreased and increased sleep durations. Data from the 2010 Community Health Survey were used in this study. We conducted logistic regression with average sleep duration as the independent variable, injury as a dependent variable, and controlling for age, sex, occupation, education, region (cities and provinces), smoking, alcohol use, body mass index, hypertension, diabetes, arthritis, and depression. Seven categories of sleep duration were established: ≤4, 5, 6, 7, 8, 9, and ≥10 hours. Using 7 hours of sleep as the reference, the adjusted injury risk (odds ratio) for those sleeping a total of ≤4 h/d was 1.53; 1.28 for 5 hours, for 1.11 for 6 hours, 0.98 for 8 hours, 1.12 for 9 hours, and 1.48 for ≥10 hours. The difference in risk was statistically significant for each category except for the 8 and 9 hours. In this study, risk increased as the sleep duration decreased or increased, except for the 8 and 9 hours. This research found that either a decrease or increase in sleep duration was associated with an increased risk for injury. The concept of proper sleep duration can be evaluated by its associated injury risk.


Loss of caspase-1 and caspase-3 protein expression in human prostate cancer.
Activation of the caspase cascade is involved in the execution of apoptosis in a variety of cellular systems. Recent studies demonstrated that caspase-1 activation was required for human prostate cancer cells to undergo apoptosis in response to transforming growth factor-beta (Y. Guo and N. Kyprianou, Cancer Res., 59: 1366-1371, 1999). In the present study, to identify the significance of caspases in prostate cancer progression, we examined the expression of three key caspases, caspase-1, caspase-3, and caspase-9, in normal and malignant human prostates. Caspase-1, -3, and -9 expression was examined at the mRNA and the protein level in a series of human normal and malignant prostate specimens. No significant differences were observed in the mRNA expression in prostatic tumors relative to the normal gland for any of the three caspases. Immunohistochemical analysis revealed that the pattern of protein expression and distribution was uniformly homogeneous in the normal prostate, with the epithelial cells exhibiting a diffuse cytoplasmic staining for caspase-1 and caspase-3. Significantly, the majority of primary prostate cancer specimens (80%) had total lack of caspase-1 immunoreactivity, whereas the remaining showed a significantly reduced expression compared with the normal prostate (P < 0.05). Caspase-3 expression was also reduced in moderately and poorly differentiated prostatic tumors compared with well-differentiated prostate adenocarcinomas and the normal prostate (P < 0.05). No significant correlation was found between the apoptotic index or Gleason grade and the pattern of caspase protein expression in the primary prostatic tumors analyzed. Western blot analysis revealed constitutive expression of the proenzyme forms of caspase-1, -3, and -9 in the human prostate cancer cell lines PC-3, DU-145, TSU-Pr1m and LNCaP, but caspase-1 expression was low in the less tumorigenic cell lines, DU-145 and LNCaP. These findings implicate the loss of caspase-1 protein as a potential step in the loss of apoptotic control during prostate tumorigenesis. This study suggests that the pattern of caspase-1 and -3 expression in prostatic tumors may have prognostic significance in disease progression.


Socio-cultural and economic antecedents of adolescent sexual decision-making and HIV-risk in rural Uganda.
With more than half of new infections occurring among youth, HIV/AIDS remains a major contributor to morbidity and mortality in Uganda. Semi-structured interviews were performed with 48 adolescents and 15 adult key informants in a rural Ugandan community to identify influences on adolescent sexual decision-making. Inductive data analytic methods revealed five thematic influences: (1) social pressure, (2) decline of the Senga (a familial figure who traditionally taught female adolescents about how to run a household), (3) cultural barriers to condom use, (4) knowledge of HIV transmission and modes of prevention, and (5) a moral injunction against sex before marriage. Influences were classified as HIV/AIDS risk and protective factors and organized to form an explanatory framework of adolescent sexual risk-taking. Risk factors pull youth toward risky behavior, while protective factors push them away. Predominance of risk over protective influences explains persistent sexual risk-taking by Ugandan youth. HIV prevention programs designed for Ugandan adolescents should take competing factors and sociocultural and economic influences into account.


Regulation of Pax3 transcriptional activity by SUMO-1-modified PML.
Pax3 is an evolutionarily conserved transcription factor that plays a major role in a variety of developmental processes. Mutations in Pax3 lead to severe malformations as seen in human Waardenburg syndrome and in the Splotch mutant mice. The transcriptional activity of Pax3 was recently shown to be repressed by Daxx whereas the oncogenic fusion protein Pax3-FKHR is unresponsive to this repressive action. Here we demonstrate that Daxx-mediated repression of Pax3 can be inhibited by the nuclear body (NB)-associated protein PML. Interestingly, this suppression of Daxx properties correlates with its recruitment to the NBs. Factors such as arsenicals and interferons that enhance NB formation, trigger both the targeting of Daxx to these nuclear structures and the relief of the repressive activity of Daxx. Conversely, lack of structurally intact NBs profoundly impairs Pax3 transcriptional activity, likely by increasing the pool of available nucleoplasmic Daxx. Moreover, a PML mutant that can not be modified by the ubiquitin-related SUMO-1 modifier is no more able to interact with Daxx. Consistently, such a mutant fails both to inhibit the Daxx repressing effect on Pax3 and to induce its accumulation into the NBs. Taken together, these results argue that SUMO-1 modified PML can derepress Pax3 transcriptional activity through sequestration of the Daxx repressor into the NBs and suggest a role for these nuclear structures in the transcriptional control by Pax proteins. Oncogene (2001) 20, 1 - 9.


Promoters from Corynebacterium glutamicum: cloning, molecular analysis and search for a consensus motif.
Relatively limited information about promoter structures in Corynebacterium glutamicum has been available until now. With the aim of isolating and characterizing such transcription initiation signals, random Sau3A fragments of C. glutamicum chromosomal DNA and of the corynebacterial phage phi GA1 were cloned into the promoter probe vector pEKplCm and selected for promoter activity by chloramphenicol resistance of transformed C. glutamicum cells. The nucleotide sequence of ten chromosomal and three phage fragments was determined and the transcriptional start (TS) sites were localized by primer extension analyses. Additionally, the promoters of five previously isolated C. glutamicum genes were cloned and mapped. All of the isolated promoters were also functional in the heterologous host Escherichia coli. A comparative analysis of the newly characterized promoter sequences together with published promoters from C. glutamicum revealed conserved sequences centred about 35 bp (ttGcca) and 10 bp (TA.aaT) upstream of the TS site. The position of these motifs and the motifs themselves are comparable to the -35 and -10 promoter consensus sequences of other Gram-positive and Gram-negative bacteria, indicating that they represent transcription initiation signals in C. glutamicum. However, the C. glutamicum consensus hexamer of the -35 region is much less conserved than in E. coli, Bacillus, Lactobacillus and Streptococcus.


Multiple versions of the A mating type locus of Coprinus cinereus are generated by three paralogous pairs of multiallelic homeobox genes.
The A mating type locus of Coprinus cinereus determines mating compatibility by regulating essential steps in sexual development. Each A locus contains several genes separated into two functionally independent complexes termed A alpha and A beta, and the multiple alleles of these genes generate an estimated 160 A mating specificities. The genes encode two classes of homeodomain-containing proteins designated HD1 and HD2. In this report we describe two newly cloned loci, A2 and A5, and compare them with A42, A43 and A6 that we have described previously. An A beta-null locus, retaining just a single active HD1 gene from the alpha-complex, was generated by mutation. Using this as a transformation host, gene combinations that promote A-regulated development were identified. We demonstrate that each A locus contains members of three paralogous pairs of HD1 and HD2 genes. Different allelic versions of gene pairs are compatible but paralogous genes are incompatible. The genes present in four uncloned A loci were deduced using Southern analyses and transformations with available cloned genes. The combined analysis of nine A factors identifies sufficient A gene alleles to generate at least 72 A mating specificities.


Recombinant human type II collagens with low and high levels of hydroxylysine and its glycosylated forms show marked differences in fibrillogenesis in vitro.
Type II collagen is the main structural component of hyaline cartilages where it forms networks of thin fibrils that differ in morphology from the much thicker fibrils of type I collagen. We studied here in vitro the formation of fibrils of pepsin-treated recombinant human type II collagen produced in insect cells. Two kinds of type II collagen preparation were used: low hydroxylysine collagen having 2.0 hydroxylysine residues/1,000 amino acids, including 1.3 glycosylated hydroxylysines; and high hydroxylysine collagen having 19 hydroxylysines/1,000 amino acids, including 8.9 glycosylated hydroxylysines. A marked difference in fibril formation was found between these two kinds of collagen preparation, in that the maximal turbidity of the former was reached within 5 min under the standard assay conditions, whereas the absorbance of the latter increased until about 600 min. The critical concentration with the latter was about 10-fold, and the absorbance/microgram collagen incorporated into the fibrils was about one-sixth. The morphology of the fibrils was also different, in that the high hydroxylysine collagen formed thin fibrils with essentially no interfibril interaction or aggregation, whereas the low hydroxylysine collagen formed thick fibrils on a background of thin ones. The data thus indicate that regulation of the extents of lysine hydroxylation and hydroxylysine glycosylation may play a major role in the regulation of collagen fibril formation and the morphology of the fibrils.


Inhibition of Aβ(1-40) fibril formation by cyclophilins.
Cyclophilins interact directly with the Alzheimer's disease peptide Aβ (amyloid β-peptide) and are therefore involved in the early stages of Alzheimer's disease. Aβ binding to CypD (cyclophilin D) induces dysfunction of human mitochondria. We found that both CypD and CypA suppress in vitro fibril formation of Aβ(1-40) at substoichiometric concentrations when present early in the aggregation process. The prototypic inhibitor CsA (cyclosporin A) of both cyclophilins as well as the new water-soluble MM258 derivative prevented this suppression. A SPOT peptide array approach and NMR titration experiments confirmed binding of Aβ(1-40) to the catalytic site of CypD mainly via residues Lys(16)-Glu(22) The peptide Aβ(16-20) representing this section showed submicromolar IC50 values for the peptidyl prolyl cis-trans isomerase activity of CypD and CypA and low-micromolar KD values in ITC experiments. Chemical cross-linking and NMR-detected hydrogen-deuterium exchange experiments revealed a shift in the populations of small Aβ(1-40) oligomers towards the monomeric species, which we investigated in the present study as being the main process of prevention of Aβ fibril formation by cyclophilins.


Thoracoscopic-assisted esophagectomy and laparoscopic gastric pull-up for lye injury.
Acquired esophageal strictures in children are often the result of ingestion of caustic agents. We describe 2 children with severe esophageal strictures following lye ingestion, who successfully underwent esophagectomy and gastric pull-up utilizing combined thoracoscopic and laparoscopic techniques. This was a retrospective chart analysis of both patients. CASE 1: A 17-year-old female, who ingested a lye-containing substance, which lead to the need for gastrostomy and esophageal dilatations, developed an esophageal stricture. Thoracoscopic esophagectomy, laparoscopic gastric conduit creation, pyloroplasty, gastric pull-up, and esophagogastric anastomosis was performed one year later. She was tolerating a regular diet for almost 4 years following esophageal replacement when she developed a gastric ulcer with gastrobronchial fistula that required open repair via a right thoracotomy. She has since recovered and resumed her regular diet. CASE 2: A 13-month-old female who ingested a lye-based cleaner underwent tracheostomy and gastrostomy on the day of injury, and esophageal dilatations beginning 1 month later. Despite dilatations, she developed severe strictures for which at age 21 months she underwent thoracoscopic esophageal mobilization, laparoscopic creation of gastric conduit, pyloroplasty, and esophagogastric anastomosis. A right thoracotomy was necessary to negotiate the conduit safely up to the neck. She is tolerating feeds and has not developed any complications for nearly 3 years following esophageal replacement. Esophagectomy and gastric pull-up for esophageal lye injuries can be accomplished utilizing a combination of thoracoscopy and laparoscopy with excellent results. Long-term follow-up is necessary to manage potential complications in these patients.


Granulocytopenia in cancer patients treated in a phase I trial with recombinant human tumor necrosis factor.
We have examined the effect of recombinant human tumor necrosis factor (TNF) upon granulocyte kinetics in cancer patients in a phase I clinical trial. TNF was given to each patient intravenously over 2 h at varying doses. A marked drop in the total white blood cell count, absolute polymorphonuclear leukocyte (PMN) count, and absolute monocyte count occurred reproducibly at 30 min after TNF initiation. Also noted was a drop in the absolute lymphocyte and eosinophil counts, both of which reached their nadir at approximately 4 h. A marked increase in immature PMN leukocytes (bands) was noted beginning at 1 h. These changes were statistically significant. Statistically significant increases in hemoglobin and hematocrit occurred at the 30 min time point but subsequently decreased to approximately 90% of pretreatment baseline. Additionally, the platelet count decreased, reaching its nadir approximately 6 h after TNF initiation. In four serial studies in patients on the highest dose of TNF, the granulocyte adhesion protein CD11b was shown to increase on the surface of the PMN leukocytes by as early as 7-15 min after initiation of TNF infusion. In each of these, expression of CD11b antigen increased prior to the disappearance of PMN leukocytes from the peripheral circulation. A similar finding was obtained for monocytes. This work indicates that within 30 min of intravenous infusion of TNF, mature granulocytes and monocytes have left the peripheral circulation. This is followed by an apparent bone marrow response indicated by an outpouring of bands. The initial granulocyte and monocyte emigration from the peripheral circulation is preceded at highest-dose TNF by increased cell surface expression of CD11b for both cell types, suggesting a causal relationship between these temporally linked events.


[Existence of a form of SV40 T antigen incapable of interacting with DNA].
The interaction of SV40 T-antigen and viral DNA was studied by using adsorption of DNA-protein complexes on nitrocellulose filters. The T-antigen purification procedure included ion-exchange chromatography on DEAE-cellulose, selective adsorption of cellular proteins on single-stranded DNA-cellulose, chromatography on heparin-Sepharose and removal of cell proteins by an immunosorbent. Only the latter step allowed to remove the contamination of cellular DNA-binding proteins, judging from the reaction of T-antigen neutralization by specific antibodies. It was shown that T-antigen and cellular DNA-binding proteins interact with SV40 DNA at different values of pH, namely ah 6,0-6,4 and 7,9, respectively. The T-antigen obtained was passed through a column with native DNA-cellulose at pH and ionic strength values optimal for interaction with DNA. The bulk of T-antigen (30-40%) did not bind to native thymus DNA and did not interact with SV40 DNA. It is assumed that this fraction is a form of T-antigen, which undergoes structural or functional changes during specific interaction with viral or cellular DNAs.


Cardiac regenerative medicine.
Severe heart failure is associated with damage to the myocardium that is irreversible with current medical therapies. Recent experimental and clinical studies, however, have opened the possibility of solving many of the associated problems, making this an exciting and tangible goal. There are many potential cell sources for regenerative cardiac medicine, including bone marrow stem cells, endothelial progenitor cells, skeletal myocytes, adult cardiac stem cells, and embryonic stem (ES) cells. Although ES cells are highly proliferative and suitable for mass production, they are not autologous, and an efficient protocol is yet to be established to ensure selective cardiomyocyte induction. Recent studies have successfully established inducible pluripotent stem (iPS) cells from mouse and human fibroblasts by the gene transfer of 4 transcription factors that are strongly expressed in ES cells: Oct3/4, Sox2, Klf4 and c-Myc. iPS cells can differentiate into all 3 germ layer-derived cells and are syngeneic, indicating that they can become an ideal cell source for regenerative medicine. Despite these successes, the accumulating evidence from fields as diverse as developmental biology, stem cell biology and tissue engineering must be integrated to achieve the full potential of cardiac regenerative medicine.


Computing the Differences between Asn-X and Gln-X Deamidation and Their Impact on Pharmaceutical and Physiological Proteins: A Theoretical Investigation Using Model Dipeptides.
Protein deamidation is a degradation mechanism that significantly impacts both pharmaceutical and physiological proteins. Deamidation impacts two amino acids, Asn and Gln, where the net neutral residues are converted into their acidic forms. While there are multiple similarities between the reaction mechanisms of the two residues, the impact of Gln deamidation has been noted to be most significant on physiological proteins while Asn deamidation has been linked to both pharmaceutical and physiological proteins. For this purpose, we sought to analyze the thermochemical and kinetic properties of the different reactions of Gln deamidation relative to Asn deamidation. In this study, we mapped the deamidation of Gln-X dipeptides into Glu-X dipeptides using density functional theory (DFT). Full network mapping facilitated the prediction of reaction selectivity between the two primary pathways, as well as between the two products of Gln-X deamidation as a function of solvent dielectric. To achieve this analysis, we studied a total of 77 dipeptide reactions per solvent dielectric (308 total reactions). Modeled at a neutral pH and using quantum chemical and statistical thermodynamic methods, we computed the following values: enthalpy of reaction (ΔHRXN), entropy (ΔSRXN), Gibbs free energy of reaction (ΔGRXN), activation energy (EA), and the Arrhenius preexponential factor (log(A)) for each dipeptide. Additionally, using chemical reaction principles, we generated a database of computed rate coefficients for all possible N-terminus Gln-X deamidation reactions at a neutral pH, predicted the most likely deamidation reaction mechanism for each dipeptide reaction, analyzed our results against our prior study on Asn-X deamidation, and matched our results against qualitative trends previously noted by experimental literature.


Exploring the evolution of the proteins of the plant nuclear envelope.
In this study, we explore the plasticity during evolution of proteins of the higher plant nuclear envelope (NE) from the most ancestral plant species to advanced angiosperms. The higher plant NE contains a functional Linker of Nucleoskeleton and Cytoskeleton (LINC) complex based on conserved Sad1-Unc84 (SUN) domain proteins and plant specific Klarsicht/Anc1/Syne homology (KASH) domain proteins. Recent evidence suggests the presence of a plant lamina underneath the inner membrane and various coiled-coil proteins have been hypothesized to be associated with it including Crowded Nuclei (CRWN; also termed LINC and NMCP), Nuclear Envelope Associated Protein (NEAP) protein families as well as the CRWN binding protein KAKU4. SUN domain proteins appear throughout with a key role for mid-SUN proteins suggested. Evolution of KASH domain proteins has resulted in increasing complexity, with some appearing in all species considered, while other KASH proteins are progressively gained during evolution. Failure to identify CRWN homologs in unicellular organisms included in the study and their presence in plants leads us to speculate that convergent evolution may have occurred in the formation of the lamina with each kingdom having new proteins such as the Lamin B receptor (LBR) and Lamin-Emerin-Man1 (LEM) domain proteins (animals) or NEAPs and KAKU4 (plants). Our data support a model in which increasing complexity at the nuclear envelope occurred through the plant lineage and suggest a key role for mid-SUN proteins as an early and essential component of the nuclear envelope.


The Effect of Chlortetracycline Meditation on the Coliform Microflora of Newly Hatched Chicks.
Newly hatched chicks were medicated with chlortetracycline at levels of 0, 400, 800 and 1200 mg. per imperial gallon of drinking water, for the first 5 days in the brooders. Coliform microflora of the ceca were counted when the chicks were first removed from the incubator and at 2 and 5 days of age. The numbers of coliforms decreased with increasing levels of the antibiotic and with increasing age. Sixtyfour per cent of the coliforms were E. coli and of these a large proportion were resistant to chlortetracycline and oxytetracycline in vitro. There was a trend towards increasing incidence of isolation of other resistant gram negative rods with increasing levels of chlortetracycline medication.


Prognostic implications of left heart diastolic dysfunction in adults with coarctation of aorta.
The prognostic implication of left atrial (LA) dysfunction and left ventricular diastolic dysfunction (LVDD) in patients with coarctation of aorta (COA) is unknown. The purpose of this study was to determine whether LA dysfunction and LVDD were associated with mortality in COA patients. This is a retrospective review of adults (age ≥18 years) with repaired COA that underwent transthoracic echocardiogram (2000-18). LVDD was determined using the 2016 guidelines for LV diastolic function assessment, and LA dysfunction was assessed using LA reservoir strain. Of 721 patients, LV diastolic function could be determined in 635 (88%); and 414 (65%) had no LVDD, while 146 (23%), 53 (8%), and 22 (4%) had Grade I/II/III LVDD, respectively. The mean LA reservoir strain was 39 ± 11%, and patients were divided into quartiles: top quartile (reference group), mild LA dysfunction, moderate LA dysfunction, and severe LA dysfunction. Grade III LVDD (but not Grades I and II) was associated with death/transplant. On the other hand, there was an incremental risk of death/transplant across LA strain quartiles: mild LA dysfunction [hazard ratio (HR) 1.16, 1.04-2.06], moderate LA dysfunction (HR 1.75, 1.27-3.58), and severe LA dysfunction (HR 3.49, 1.88-7.16). Of 86 patients with indeterminate diastolic function, there was a trend towards a lower 5-year transplant-free survival in patients with LA dysfunction vs. normal LA function (83% vs. 91%, P = 0.06). LA dysfunction (but not LVDD) was associated with incremental risk of mortality and thus can be used for prognostication in all patients including those with indeterminate diastolic function.


Glucose-, calcium- and concentration-dependence of acetylcholine stimulation of insulin release and ionic fluxes in mouse islets.
Mouse islets were used to define the glucose-dependence and extracellular Ca2+ requirement of muscarinic stimulation of pancreatic beta-cells. In the presence of a stimulatory concentration of glucose (10 mM) and of Ca2+, acetylcholine (0.1-100 microM) accelerated 3H efflux from islets preloaded with myo-[3H]inositol. It also stimulated 45Ca2+ influx and efflux, 86Rb+ efflux and insulin release. In the absence of Ca2+, only 10-100 microM-acetylcholine mobilized enough intracellular Ca2+ to trigger an early but brief peak of insulin release. At a non-stimulatory concentration of glucose (3 mM), 1 microM- and 100 microM-acetylcholine increased 45Ca2+ and 86Rb+ efflux in the presence and absence of extracellular Ca2+. However, only 100 microM-acetylcholine marginally increased 45Ca2+ influx and caused a small, delayed, stimulation of insulin release, which was abolished by omission of Ca2+. At a maximally effective concentration of glucose (30 mM), 1 microM- and 100 microM-acetylcholine increased 45Ca2+ influx and efflux only slightly, but markedly amplified insulin release. Again, only 100 microM-acetylcholine mobilized enough Ca2+ to trigger a peak of insulin release in the absence of Ca2+. The results thus show that only high concentrations of acetylcholine (greater than or equal to 10 microM) can induce release at low glucose or in a Ca2+-free medium. beta-Cells exhibit their highest sensitivity to acetylcholine in the presence of Ca2+ and stimulatory glucose. Under these physiological conditions, the large amplification of insulin release appears to be the result of combined effects of the neurotransmitter on Ca2+ influx, on intracellular Ca2+ stores and on the efficiency with which Ca2+ activates the releasing machinery.


An improved light dark box test by using a real-time video tracking system.
In the behavioral science field, many of the oldest tests have still most frequently been used almost in the same way for decades. The subjective influence of human observer and the large inter-observer and interlab differences are substantial among these tests. This necessitates the possibility of using technological innovations for behavioral science to obtain new parameters, results and insights as well. The light-dark box (LDB) test is a characteristic tool used to assess anxiety in rodents. A complete behavioral analysis (including both anxiety and locomotion parameters) is not possible by performing traditional LDB test protocol, as it lacks the usage of a real-time video recording of the test. In the current report, we describe an improved approach to conduct LDB test using a real-time video tracking system.


The connected health of cardiovascular medicine: current status and future directions.
The technologies of cardiovascular connected health stand to dramatically alter the management and prevention of cardiovascular disease, a worldwide leading cause of death. The American Heart Association has outlined seven key health metrics including physical activity, adequate blood pressure control, weight and a healthy diet, which lie at the core of cardiovascular disease management. Controlling these metrics has been demonstrated to result in substantial reductions in cardiovascular mortality. These metrics are ideally suited to a connected health management strategy involving enhanced patient empowerment and augmented physician engagement. As more patients and healthcare providers adopt technologies that allow for self-monitoring and point-of-care diagnostics, the physician has access to a greater depth of data concerning their patient's health and how best to influence it.


Personality change predicts self-reported mental and physical health.
Personality dimensions are known to predict mortality and other health outcomes, but almost no research has assessed the effects of changes in personality traits on physical and mental health outcomes. In this article, we examined the effects of changes in the Big Five personality dimensions on health as assessed by the Short Form Health Survey (SF-36). Respondents were 11,105 Australian adults aged 20-79 years (52.7% female). Latent difference score modeling was used to examine whether personality change over a 4-year period was associated with mental and physical health, and whether these effects were moderated by birth cohort. Increases in Conscientiousness and Extraversion were found to be associated with improved mental and physical health, whereas increased Neuroticism was linked with poorer health. The nature of these associations varied significantly by birth cohort. The findings have implications for understanding how changes in personality traits over time are related to health, and could be used to aid the development of effective health promotion strategies targeted to specific personality traits and birth cohorts.


Continuous interscalene block in patients having outpatient rotator cuff repair surgery: a prospective randomized trial.
We performed this randomized trial to compare the recovery profile of patients receiving single injection (SISB) and continuous interscalene brachial plexus block (CISB) or general anesthesia (GA) for arthroscopic rotator cuff repair surgery through the first postoperative week. Our primary hypothesis was that the highest pain numeric rating scale (NRS) (worst pain score) at the end of the study week would be lower for patients in the CISB group than for patients in the SISB or GA groups. Seventy-one patients scheduled for elective outpatient arthroscopic rotator cuff repair were enrolled. CISB patients received 20 mL of 0.5% ropivacaine as a bolus through a catheter, whereas SISB patients received the same injection volume through a needle. CISB patients received an infusion of 0.2% ropivacaine at 5 mL/h with a patient-controlled bolus of 5 mL hourly for 48 hours. GA-only patients received a standardized general anesthetic. Postoperative highest NRS pain scores through the first postoperative week, time-to-first pain, analgesic consumption, fast-tracked postoperative anesthesia care unit (PACU) bypass rate, length of PACU stay, time-to-discharge home, total hours of sleep, and related adverse effects were recorded in the PACU and at home on postoperative days 1, 2, 3, and 7. No patient in the CISB or SISB groups reported a NRS ≥1 or required analgesics while in the PACU. While most patients in the CISB and SISB groups were fast-tracked to PACU discharge, no patient in the GA group was fast-tracked (Χ P = 0.003). Length of stay in the PACU was significantly shorter for the CISB and SISB groups than for the GA group (20 ± 31, 30 ± 42, and 165 ± 118 minutes, respectively (CISB vs GA, P < 0.001; SISB vs GA, P <0.001), and time-to-discharge home was significantly shorter when compared with the GA group. Time to first pain report was longer in the CISB group. Mean NRS scores were lower for patients in the CISB group than in the SISB and GA groups on postoperative days 1 and 2, and use of narcotics (doses ≥1) was lower until postoperative day 3. Patients who received CISB slept significantly longer than patients who received SISB or GA (P < 0.01) during the first 48 hours postoperatively. By the end of the study week, 26% of patients in the CISB group, 83% in the SISB group, and 58% of GA patients reported NRS ≥4 (both P-values ≤ 0.05). The analgesic benefits of CISB found in the PACU and immediately after discharge extend through the intermediate recovery period ending on postoperative day 7.


Classifying patients by antipsychotic adherence patterns using latent class analysis: characteristics of nonadherent groups in the California Medicaid (Medi-Cal) program.
This study identifies latent classes defined by varying degrees of adherence to antipsychotic drug therapy and examines the sociodemographic, clinical, and resource utilization correlates associated with membership in each adherence class. Patient-level data were drawn from the 1994 to 2003, 100%-sample California Medicaid fee-for-service paid claims data for patients with schizophrenia (N = 36,195). The date of the first antipsychotic medication filled after January 1, 1999 was then used to divide each patient's data into a 6-month preindex (baseline) and a 12-month postindex (follow-up) period. Three categorical adherence indicators-a dichotomous variable of medication possession ratio greater than 0.80, the number of antipsychotic treatment attempts, and time to a change in antipsychotic medications-and two covariates-a categorical variable of duration of therapy and a dichotomous variable of polypharmacy-were used in the latent class model. A three-class model returned the lowest values for all the information criteria and was therefore interpreted as follows: The prevalence rates of the latent classes were 1) 14.8% for the adherent; 2) 20.7% for the partially adherent; and 3) 64.5% for the nonadherent. Membership in the nonadherent class was associated with minority ethnicity, being female, eligibility due to welfare status, prior hospitalizations, and a higher number of prior treatment episodes. Membership in the partially adherent class was associated with higher use of outpatient care, higher rates of depot antipsychotic drug use, and polypharmacy. Multiple indicators of adherence to antipsychotic medication can be used to define classes of adherence that are associated with patient characteristics and distinct patterns of prior health-care use.


A land use regression model of nitrogen dioxide and fine particulate matter in a complex urban core in Lanzhou, China.
Land use regression (LUR) models have been widely used to estimate air pollution exposures at high spatial resolution. However, few LUR models were developed for rapidly developing urban cores, which have substantially higher densities of population and built-up areas than the surrounding areas within a city's administrative boundary. Further, few studies incorporated vertical variations of air pollution in exposure assessment, which might be important to estimate exposures for people living in high-rise buildings. A LUR model was developed for the urban core of Lanzhou, China, along with a model of vertical concentration gradients in high-rise buildings. In each of four seasons in 2016-2017, NO2 was measured using Ogawa badges for 2 weeks at 75 ground-level sites. PM2.5 was measured using DataRAM for shorter time intervals at a subset (N = 38) of the 75 sites. Vertical profile measurements were conducted on 9 stories at 2 high-rise buildings (N = 18), with one building facing traffic and another facing away from traffic. The average seasonal concentrations of NO2 and PM2.5 at ground level were regressed against spatial predictors, including elevation, population, road network, land cover, and land use. The vertical variations were investigated and linked to ground-level predictions with exponential models. We developed robust LUR models at the ground level for estimated annual averages of NO2 (R2: 0.71, adjusted R2: 0.67, and Leave-One-Out Cross Validation (LOOCV) R2: 0.64) and PM2.5 (R2: 0.77, adjusted R2: of 0.73, and LOOCV R2: 0.67) in the urban core of Lanzhou, China. The LUR models for the estimated seasonal averages of NO2 showed similar patterns. Vertical variation of NO2 and PM2.5 differed by windows orientation with respect to traffic, by season or by time of a day. Vertical variation functions incorporated the ground-level LUR predictions, in a form that could allow for exposure assessment in future epidemiological investigations. Ground-level NO2 and PM2.5 showed substantial spatial variations, explained by traffic and land use patterns. Further, vertical variation of air pollution levels is significant under certain conditions, suggesting that exposure misclassification could occur with traditional LUR that ignores vertical variation. More studies are needed to fully characterize three-dimensional concentration patterns to accurately estimate air pollution exposures for residents in high-rise buildings, but our LUR models reinforce that concentration heterogeneity is not captured by the limited government monitors in the Lanzhou urban area.


spp42, identified as a classical suppressor of prp4-73, which encodes a kinase involved in pre-mRNA splicing in fission yeast, is a homologue of the splicing factor Prp8p.
We have identified two classical extragenic suppressors, spp41 and spp42, of the temperature sensitive (ts) allele prp4-73. The prp4(+) gene of Schizosaccharomyces pombe encodes a protein kinase. Mutations in both suppressor genes suppress the growth and the pre-mRNA splicing defect of prp4-73(ts) at the restrictive temperature (36 degrees ). spp41 and spp42 are synthetically lethal with each other in the presence of prp4-73(ts), indicating a functional relationship between spp41 and spp42. The suppressor genes were mapped on the left arm of chromosome I proximal to the his6 gene. Based on our mapping data we isolated spp42 by screening PCR fragments for functional complementation of the prp4-73(ts) mutant at the restrictive temperature. spp42 encodes a large protein (p275), which is the homologue of Prp8p. This protein has been shown in budding yeast and mammalian cells to be a bona fide pre-mRNA splicing factor. Taken together with other recent genetic and biochemical data, our results suggest that Prp4 kinase plays an important role in the formation of catalytic spliceosomes.


Incorporating prior information with fused sparse group lasso: Application to prediction of clinical measures from neuroimages.
Predicting clinical variables from whole-brain neuroimages is a high-dimensional problem that can potentially benefit from feature selection or extraction. Penalized regression is a popular embedded feature selection method for high-dimensional data. For neuroimaging applications, spatial regularization using the ℓ1 or ℓ2 norm of the image gradient has shown good performance, yielding smooth solutions in spatially contiguous brain regions. Enormous resources have been devoted to establishing structural and functional brain connectivity networks that can be used to define spatially distributed yet related groups of voxels. We propose using the fused sparse group lasso (FSGL) penalty to encourage structured, sparse, and interpretable solutions by incorporating prior information about spatial and group structure among voxels. We present optimization steps for FSGL penalized regression using the alternating direction method of multipliers algorithm. With simulation studies and in application to real functional magnetic resonance imaging data from the Autism Brain Imaging Data Exchange, we demonstrate conditions under which fusion and group penalty terms together outperform either of them alone.


Predicting nursing facility transition candidates using AID: a case study.
Although the nursing facility transition literature is growing, little research has analyzed the characteristics of individuals so assisted or compared participants to those who remain institutionalized. This article describes an analytic method that researchers can apply to address these knowledge gaps, using the Arkansas Passages nursing facility transition program as a case study. This study employed Arkansas Minimum Data Set 2.0 data for 111 transitioned individuals, a derivation sample of 1,000 other residents, and a validation sample of all residents from the transitioned individuals' nursing facilities. Tree classification techniques identified distinct groups of transitioned and nontransitioned residents. Nearly two thirds of transitionees were part of a group comprising only 1.5% of all Arkansas nursing facility residents. Five characteristics identified this group: age, day of stay (i.e., current day of stay at the time of the assessment), having hemiplegia/paraplegia, cognitive impairment level, and classification into one of eight Resource Utilization Groups (RUG-III) case-mix groups associated with the least nursing staff time. Another group containing 92% of the transitionees comprised 22% of all residents. Two characteristics identified this group: being younger than age 65 or being in the eight low-resource RUG-III groups. Given that the majority of individuals assisted by this pilot represent a small and unusual nursing facility subpopulation, policy makers may wish to exercise caution in utilizing these data to forecast future transition populations, costs, or outcomes. Replicating this analysis using additional states' data could increase understanding about the characteristics of those assisted across transition programs and could help construct a more robust definition of what constitutes a transition success.


Airflow and patient-perceived improvement following rhinoplastic correction of external nasal valve dysfunction.
External nasal valve dysfunction (ENVD) is a common cause of nasal obstruction. Although many techniques are described to help correct ENVD, evidence of the objective changes in the airway achieved by these interventions is mainly unknown. To document the airway changes in patients with ENVD by comparing subjective and objective measures obtained before and after rhinoplasty. Prospective case series with validated subjective and objective outcomes at a tertiary rhinologic center in Sydney, Australia. We included 19 patients with nasal obstruction and clinically diagnosed ENVD from January 2012 to May 2013. Functional reconstructive rhinoplasty involving lateral crural underlay strut grafts using costal cartilage or lateral crural cephalic turn-in maneuvers performed to correct ENVD. Objective assessment included nasal peak inspiratory flow, nasal airway resistance, and minimum cross-sectional area. Subjective assessment included a visual analog scale for nasal obstruction, the 22-item Sinonasal Outcome Test, the Nasal Obstruction Symptom Evaluation Scale, and the 36-Item Short Form Health Survey, version 2. A 13-point Likert scale was also used to assess overall function and cosmesis. Objective data and visual analog scale scores were obtained before and after decongestion at baseline and 6 months after surgery. Mean (SD) age of the patients undergoing assessment was 33.3 (12.4) years; 13 patients (68%) were female. Significant improvement was observed in scores for the Sinonasal Outcome Test (mean [SD] change, 0.85 [0.96]), Nasal Obstruction Symptom Evaluation Scale (mean [SD] change, 30.53 [26.14]), and overall function (median [25th-75th percentiles] change, -6.5 [-7.0 to 1.0]) and cosmesis (median [25th-75th percentiles] change, -4.0 [-8.0 to -1.0]) (P < .01). The mean (SD) nasal peak inspiratory flow increased from 102.6 (45.6) to 124.0 (52.9) L/min (P < .01). Median (25th-75th percentiles) nasal airway resistance showed no significant change (from 0.296 [0.237-0.414] to 0.292 [0.267-0.371] Pa/cm3/s; P = .92). The minimum cross-sectional area also showed no significant change (mean [SD], from 1.188 [0.407] to 1.229 [0.336] cm2; P = .69). Contrary to common belief, successful rhinoplasty had little effect on structural shape or resistance in ENVD, but symptoms improved with changes in collapsibility as defined by the nasal peak inspiratory flow. The need to reconstruct lateral wall support is reinforced by the data presented. 4.


Novel strategies exploiting interleukin-12 in cancer immunotherapy.
Interleukin-12 is considered a potent agent to enhance antitumor immune responses. It belongs to a family of heterodimeric cytokines with key roles in the up-regulation and down-regulation of cellular immunity. Since its discovery, recombinant IL-12 was found to exert potent antitumor effects in rodent tumor models and was rapidly tested in the clinic with an unfavorable benefit/toxicity profile. Localized delivery of IL-12 dramatically improves the therapeutic index and this approach is being applied in the clinic based on in-vivo electroporation of naked plasmid DNA encoding IL-12, mRNA formulations, viral vectors and tumor-targeted fusion proteins. Other biotechnology strategies such as IL-12-engineered local adoptive cell therapy and pro-cytokines can also be used to improve results and broaden the therapeutic window. Combination strategies of such localized IL-12-based approaches with checkpoint inhibitors are yielding promising results both preclinically and in the early-phase clinical trials.


Dominance behavior in the rat following lesions of the caudate nuclei.
Rats with bilateral lesions of the anterior caudate nucleus or the dorsolateral frontal cortex were tested against sham-operated rats in a straight-alley dominance situation. In 91% of encounters the caudate lesion rats forced the sham-operated animals to retreat. The frontal lesion rats won only 56% of their encounters, a proportion no different to that expected by chance. The performance of the caudate-damaged rats could not be explained satisfactorily in terms of increased motivation, changes in activity, motor impairments, or perseverative tendencies. It seemed more likely that the apparent increase in dominance was associated with changes in the level of aggression.


Continuous quality improvement at work: the first team--Part II.
This second part of a two-part article follows Cape Canaveral Hospital's first continuous quality improvement team through the processes of goal setting, system analysis, data gathering, and problem resolution in the area of patients' assignment to observation status. The team's primary goal was data-driven improvement. As detailed here, the team's solution to improve the use of observation status is both time-efficient and offers opportunities for financial gain.


Obesity, Treatment Times, and Cardiovascular Outcomes After ST-Elevation Myocardial Infarction: Findings From Mission: Lifeline North Texas.
With increasing rates of obesity and its link with cardiovascular disease, there is a need for better understanding of the obesity-outcome relationship. This study explores the association between categories of obesity with treatment times and mortality for patients experiencing ST-segment elevation myocardial infarction. We examined 8725 patients with ST-segment elevation myocardial infarction who underwent primary percutaneous coronary intervention and used regression models to analyze the relationship between 6 categories of body mass index with key door-to-balloon time, total ischemic time, and in-hospital mortality. We relied on data from the Mission: Lifeline North Texas program, consisting of 33 percutaneous coronary intervention-capable hospitals in 6 counties surrounding Dallas, Texas. Data were extracted from the National Cardiovascular Data Registry for each participating hospital. Of the samples, 76% were overweight or obese. Comparing the univariate differences between the normal-weight group and the pooled sample, we observed a U-shaped association between body mass index and both mortality and door-to-balloon times. The most underweight and severely obese had the highest mortality and median door-to-balloon time, respectively. These differences persisted after multivariate adjustments for door-to-balloon time, but not for mortality. Extremely obese patients have longer treatment time delays than other body mass index categories. However, this did not extend to significant differences in mortality in the multivariate models. We conclude that clinicians should incorporate body mass assessments into their diagnosis and treatment plans to mitigate observed disparities.


[Substernal goiters].
In a series of 363 patients operated for goiter, 60 were retrosternal (16.5%). The female incidence although less than in cervical goiters remains predominant (2,5:1). Most patients are in older age group, 68% being above 40 years. The majority of goiters are multinodular and of long duration. Bilateral (37%) and left sided location (43%) were common. The incidence of cancer was 7%, thyrotoxicosis was noted in 5 patients (8%). Most patients were symptomatic (97%). Diagnosis is easily done by physical examination, chest X-Ray and thyroid scintigraphy. Computed tomography might be helpful. Retrosternal goiter is an indication to surgery except in high risk patients. The cervical approach has been used in 98% of cases. There were no post-operative death and no major complications (compressive hematoma, laryngeal nerve paralysis, hypocalcemia).


Differential expression of liver interleukin-6 receptor-alpha in female versus male ethanol-consuming rats.
It is well known that women are more susceptible to alcoholic liver disease (ALD) than men, and inflammation is thought to play a major role in alcohol-induced liver injury. Increased circulating levels of the proinflammatory cytokine interleukin (IL)-6 are a marker for serious ALD in humans. However, IL-6 also has protective effects, such as induction of liver regeneration and inhibition of hepatocyte apoptosis. Although the roles of IL-6 in ALD have begun to be established, little is known about the expression of its receptor (IL-6Ralpha) during chronic alcohol administration. Male and female rats were intragastrically fed ethanol or control isocaloric liquid diet for 2 and 4 weeks. Liver samples were collected, and gene expression was assessed by reverse transcription-polymerase chain reaction and Western blot. Herein, we show clear gender differences in alcohol-induced liver IL-6Ralpha expression. Analysis of rat liver samples showed that ethanol consumption significantly increased IL-6Ralpha messenger RNA and protein expression in females as compared with similarly treated males after 2 and 4 weeks. Increased STAT3 phosphorylation in the livers of ethanol-consuming females also indicated greater IL-6Ralpha activation in these animals. Conversely, ethanol-consuming males displayed increased IkappaB messenger RNA and protein expression, which may inhibit IL-6R expression, compared with females. Given the association of inflammation with ethanol-induced liver damage, these data may offer insight into a possible mechanism by which females develop more severe ALD than males.


Aetiological profile of paediatric stridor in a Malaysian tertiary hospital.
Conditions causing stridor in paediatric patients can range from minor illnesses to life-threatening disorders. Proper evaluation and correct diagnosis are essential for timely intervention. The objective of this study was to determine the aetiological profiles and the management of paediatric patients with stridor referred to the Otorhinolaryngology Department of Hospital Serdang. Medical records of all paediatric patients presenting with symptom of stridor from January 2010 to February 2015 were reviewed retrospectively. The patients' demographic data, clinical notes, laryngoscope findings, diagnosis and management were retrieved and analysed. Out of the total 137 patients referred for noisy breathing, 121 patients had stridor and were included in this study. There were 73 males and 48 females-most were of Malay ethnicity (77.7%). The age of presentation ranged from newborn to 10 years, with a mean of 4.9 months. Eighteen patients (14.9%) had associated congenital pathologies. The majority were congenital causes (90.9%), in which laryngomalacia was the commonest (78.5%), followed by subglottic stenosis (5.0%), vallecular cyst (2.5%) and congenital vocal fold paralysis (2.5%). Twelve patients (9.9%) had synchronous airway lesion. The majority of the patients were managed conservatively. Thirty-one patients (25.6%) required surgical intervention, of which only one needed tracheostomy. Laryngomalacia was the commonest cause of stridor among paediatric patients. A synchronous airway lesion should be considered if the child has persistent or severe symptoms. The majority of the patients were managed conservatively.


Television Watching and Telomere Length Among Adults in Southwest China.
To explore the independent associations of sedentary behavior and physical activity with telomere length among Chinese adults. Data on total time of sedentary behavior, screen-based sedentary behavior (including television watching and computer or phone use), moderate to vigorous physical activity, and dietary intake of 518 adults in Chengdu, Guizhou, and Xiamen in China (54.25% women) aged 20 to 70 years were obtained between 2013 and 2015 through questionnaires. Height, weight, and waist circumference were measured to calculate body mass index and percentage of body fat. Telomere length was measured through Southern blot technique. Television watching was inversely related to adjusted telomere length (-71.75 base pair; SE = 34.40; P = .04). Furthermore, a similar trend between telomere length and television watching was found in the group aged 20 to 40 years after adjusting for all covariates. Adults aged 20 to 40 years in the highest tertile of daily time spent on watching television had 4.0% shorter telomere length than adults in the lowest tertile (P = .03). Although the association is modest, television watching is inversely related to telomere length among Chinese adults, warranting further investigation in large prospective studies.


Qualitative aspects and validation of a screening method for pesticides in vegetables and fruits based on liquid chromatography coupled to full scan high resolution (Orbitrap) mass spectrometry.
The analytical capabilities of liquid chromatography with single-stage high-resolution mass spectrometry have been investigated with emphasis on qualitative aspects related to selective detection during screening and to identification. The study involved 21 different vegetable and fruit commodities, a screening database of 556 pesticides for evaluation of false positives, and a test set of 130 pesticides spiked to the commodities at 0.01, 0.05, and 0.20 mg/kg for evaluation of false negatives. The final method involved a QuEChERS-based sample preparation (without dSPE clean up) and full scan acquisition using alternating scan events without/with fragmentation, at a resolving power of 50,000. Analyte detection was based on extraction of the exact mass (±5 ppm) of the major adduct ion at the database retention time ±30 s and the presence of a second diagnostic ion. Various options for the additional ion were investigated and compared (other adduct ions, M + 1 or M + 2 isotopes, fragments). The two-ion approach for selective detection of the pesticides in the full scan data was compared with two alternative approaches based on response thresholds. Using the two-ion approach, the number of false positives out of 11,676 pesticide/commodity combinations targeted was 36 (0.3 %). The percentage of false negatives, assessed for 2,730 pesticide/commodity combinations, was 13 %, 3 %, and 1 % at the 0.01-, 0.05-, and 0.20-mg/kg level, respectively (slightly higher with fully automated detection). Following the SANCO/12495/2011 protocol for validation of screening methods, the screening detection limit was determined for 130 pesticides and found to be 0.01, 0.05, and ≥0.20 mg/kg for 86, 30, and 14 pesticides, respectively. For the detected pesticides in the spiked samples, the ability for unambiguous identification according to EU criteria was evaluated. A proposal for adaption of the criteria was made.


Developmental Programming of Obesity and Liver Metabolism by Maternal Perinatal Nutrition Involves the Melanocortin System.
Maternal obesity predisposes offspring to metabolic dysfunction and Non-Alcoholic Fatty Liver Disease (NAFLD). Melanocortin-4 receptor (Mc4r)-deficient mouse models exhibit obesity during adulthood. Here, we aim to determine the influence of the Mc4r gene on the liver of mice subjected to perinatal diet-induced obesity. Female mice heterozygous for Mc4r fed an obesogenic or a control diet for 5 weeks were mated with heterozygous males, with the same diet continued throughout pregnancy and lactation, generating four offspring groups: control wild type (C_wt), control knockout (C_KO), obese wild type (Ob_wt), and obese knockout (Ob_KO). At 21 days, offspring were genotyped, weaned onto a control diet, and sacrificed at 6 months old. Offspring phenotypic characteristics, plasma biochemical profile, liver histology, and hepatic gene expression were analyzed. Mc4r_ko offspring showed higher body, liver and adipose tissue weights respect to the wild type animals. Histological examination showed mild hepatic steatosis in offspring group C_KO. The expression of hepatic genes involved in regulating inflammation, fibrosis, and immune cell infiltration were upregulated by the absence of the Mc4r gene. These results demonstrate that maternal obesogenic feeding during the perinatal period programs offspring obesity development with involvement of the Mc4r system.


Analyzing Differential Regulatory Networks Modulated by Continuous-State Genomic Features in Glioblastoma Multiforme.
Gene regulatory networks are a global representation of complex interactions between molecules that dictate cellular behavior. Study of a regulatory network modulated by single or multiple modulators' expression levels, including microRNAs (miRNAs) and transcription factors (TFs), in different conditions can further reveal the modulators' roles in diseases such as cancers. Existing computational methods for identifying such modulated regulatory networks are typically carried out by comparing groups of samples dichotomized with respect to the modulator status, ignoring the fact that most biological features are intrinsically continuous variables. Here, we devised a sliding window-based regression scheme and proposed the Regression-based Inference of Modulation (RIM) algorithm to infer the dynamic gene regulation modulated by continuous-state modulators. We demonstrated the improvement in performance as well as computation efficiency achieved by RIM. Applying RIM to genome-wide expression profiles of 520 glioblastoma multiforme (GBM) tumors, we investigated miRNA- and TF-modulated gene regulatory networks and showed their association with dynamic cellular processes and brain-related functions in GBM. Overall, the proposed algorithm provides an efficient and robust scheme for comprehensively studying modulated gene regulatory networks.


Clinicopathologic significance of in vivo antinuclear autoantibodies in oral mucosal biopsies.
Although antinuclear autoantibody (ANA) staining of oral biopsy specimens is indicative of chronic ulcerative stomatitis, it is not known whether this staining is characteristic of other autoimmune diseases. Our study was undertaken to characterize the various in vivo ANA patterns detected in the oral mucosa by direct immunofluorescence to describe the associated hematoxylin and eosin findings, and determine whether patients with these findings had a coexisting systemic connective tissue disease. This was a retrospective analysis of oral biopsy specimens submitted from 2013 to 2016. In vivo ANA staining was present in 72 of the 2019 cases examined. Immunoglobulin G was the most common immunoreactant (71 of 72 cases), and speckled nuclear staining was the most frequent in vivo ANA pattern (52 of 72). In most cases, hematoxylin and eosin staining of biopsy specimens showed mucositis (24 of 34). Detailed clinical information was available for 10 patients, and all of them had an autoimmune disease. We found similar prevalence of ANA staining with direct immunofluorescence in oral epithelial biopsy specimens as reported for those of skin. In vivo ANA in the oral epithelium may indicate the presence of an immune-mediated disease. Patients who show ANA deposits in oral mucosal biopsy specimens should be investigated for systemic connective tissue disease as well as for chronic ulcerative stomatitis.


Challenges in the diagnosis of dementia: insights from the United Kingdom-Brazil Dementia Workshop.
In July 2019, a group of multidisciplinary dementia researchers from Brazil and the United Kingdom (UK) met in the city of Belo Horizonte, Minas Gerais, Brazil, to discuss and propose solutions to current challenges faced in the diagnosis, public perception and care of dementia. Here we summarize the outcomes from the workshop addressing challenges in diagnosis. Brazil faces a major problem in dementia underdiagnosis, particularly involving the population in an adverse socioeconomic context. There is poor availability of resources and specialists, and the knowledge of general practitioners and other healthcare professionals is far from satisfactory. Low education level is a further obstacle in diagnosing dementia, as the most commonly used screening tests are not designed to evaluate this population. Patients and their families must overcome the stigma of a diagnosis of dementia, which is still prevalent in Brazil and increases the burden of this condition. Whilst the UK has greater resources, dedicated memory services and a National Dementia Strategy plan, the National Health Service (NHS) has limited funding. Therefore, some challenges regarding diagnosis are common across both countries. The authors suggest possible solutions to confront these, with the goal of improving assessment and recognition of dementia and reducing misdiagnosis.


Cognition, academic achievement, and adaptive behavior in school-aged girls with fragile X syndrome.
Fragile X syndrome (FXS) is the leading monogenic cause of intellectual disability and autism in males and females. Females with FXS typically display a milder cognitive phenotype than males, despite experiencing significant developmental, behavioral, and social-emotional issues. To measure and distinguish the cognitive-behavioral profile of girls with FXS relative to verbal IQ-matched peers. Ninety-seven participants (NFXS=55, Ncomparison=42) six to 16 years of age completed assessments evaluating cognition, academic achievement, and adaptive behavior. The comparison group consisted of age-, sex-, and verbal IQ-matched peers. Consistent with previous studies, the FXS group demonstrated mean cognitive skills, academic achievement, and adaptive behavior in the borderline to low average range. On average, the FXS group showed poorer nonverbal reasoning, visual pattern recognition, verbal abstraction, math abilities, attention, inhibitory control, and working memory than the comparison group. There were no significant group differences in adaptive behavior. Different patterns of associations between cognition and selected outcomes emerged in each group. Results highlight the importance of identifying specific cognitive-behavioral profiles in girls with FXS to inform more targeted interventions for optimizing outcomes and quality of life in this population.


High-Density Lipoproteins Are Bug Scavengers.
Lipoproteins were initially defined according to their composition (lipids and proteins) and classified according to their density (from very low- to high-density lipoproteins-HDLs). Whereas their capacity to transport hydrophobic lipids in a hydrophilic environment (plasma) is not questionable, their primitive function of cholesterol transporter could be challenged. All lipoproteins are reported to bind and potentially neutralize bacterial lipopolysaccharides (LPS); this is particularly true for HDL particles. In addition, HDL levels are drastically decreased under infectious conditions such as sepsis, suggesting a potential role in the clearance of bacterial material and, particularly, LPS. Moreover, "omics" technologies have unveiled significant changes in HDL composition in different inflammatory states, ranging from acute inflammation occurring during septic shock to low-grade inflammation associated with moderate endotoxemia such as periodontal disease or obesity. In this review, we will discuss HDL modifications associated with exposure to pathogens including bacteria, viruses and parasites, with a special focus on sepsis and the potential of HDL therapy in this context. Low-grade inflammation associated with atherosclerosis, periodontitis or metabolic syndrome may also highlight the protective role of HDLs in theses pathologies by other mechanisms than the reverse transport of cholesterol.


Sensitivity analysis and power for instrumental variable studies.
In observational studies to estimate treatment effects, unmeasured confounding is often a concern. The instrumental variable (IV) method can control for unmeasured confounding when there is a valid IV. To be a valid IV, a variable needs to be independent of unmeasured confounders and only affect the outcome through affecting the treatment. When applying the IV method, there is often concern that a putative IV is invalid to some degree. We present an approach to sensitivity analysis for the IV method which examines the sensitivity of inferences to violations of IV validity. Specifically, we consider sensitivity when the magnitude of association between the putative IV and the unmeasured confounders and the direct effect of the IV on the outcome are limited in magnitude by a sensitivity parameter. Our approach is based on extending the Anderson-Rubin test and is valid regardless of the strength of the instrument. A power formula for this sensitivity analysis is presented. We illustrate its usage via examples about Mendelian randomization studies and its implications via a comparison of using rare versus common genetic variants as instruments.


Accessibility and acceptability of the Department of Veteran Affairs health care: diverse veterans' perspectives.
Diverse veteran's perspectives on the accessibility and acceptability of the Department of Veteran Affairs (VA) health services are presented. The qualitative methodology uses 16 focus groups (N = 178) stratified by war cohort (World War II and Korean Conflict versus Vietnam War and Persian Gulf War) and four ethnic/racial categories (African American, Asian American, European American, Hispanic American). Five themes emerged regarding veterans' health care expectations: (1) better information regarding available services, (2) sense of deserved benefits, (3) concern about welfare stigma, (4) importance of physician attentiveness, and (5) staff respect for patients as veterans. Although veterans' ethnic/racial backgrounds differentiated their military experiences, it was the informants' veteran identity that framed what they expected of VA health services. Accessibility and acceptability of VA health care is related to veterans' perspectives of the nature of their entitlement to service. Provider education and customer service strategies should consider the identified factors to increase access to VA as well as improve veterans' acceptance of the care.


Liver, NAFLD and COVID-19.
Coronavirus disease 2019 (COVID-19) is characterized by a wide clinical spectrum that includes abnormalities in liver function indicative of liver damage. Conversely, people with liver diseases are at higher risk of severe COVID-19. In the current review, we summarize first the epidemiologic evidence describing the bidirectional relationship between COVID-19 and liver function/liver diseases. Additionally, we present the most frequent histologic findings as well as the most important direct and indirect mechanisms supporting a COVID-19 mediated liver injury. Furthermore, we focus on the most frequent liver disease in the general population, non-alcoholic or metabolic-associated fatty liver disease (NAFLD/MAFLD), and describe how COVID-19 may affect NAFLD/MAFLD development and progression and conversely how NAFLD/MAFLD may further aggravate a COVID-19 infection. Finally, we present the long-term consequences of the pandemic on the development and management of NAFLD.


Drug resistance in Plasmodium: natural products in the fight against malaria.
Malaria, perhaps one of the most serious and widespread diseases encountered by mankind, continues to be a major threat to about 40 % of the world's population, especially in the developing world. As malaria vaccines remain problematic, chemotherapy still is the most important weapon in the fight against the disease. However, almost all available drugs have been compromised by the highly adaptable parasite, and the increasing drug resistance of Plasmodium falciparum continues to be the main problem. Therefore, the limited clinical repertoire of effective drugs and the emergence of multi-resistant strains substantiate the need for new anti-malarials. Plant-derived artemisinin is currently the only available drug that is globally effective, but alarmingly, recent studies suggest that resistance already may be developing. Nevertheless, the success story of artemisinin from the herb Qing Hao (Artemisia annua L.), used as a remedy in traditional Chinese medicine for more than two thousand years, shows once again that natural products serve as an invaluable reservoir of lead compounds for sophisticated small molecules. This review outlines the major anti-malarials, summarizing recent knowledge about their mode of action and the development of drug resistance. Furthermore, the most promising and recently discovered natural products with anti-malarial potential will be introduced.


Effects of flow-rate, inoculum and time on the internal resistance of microbial fuel cells.
To process large volumes of wastewater, microbial fuel cells (MFCs) would require anodophilic bacteria preferably operating at high flow-rates. The effect of flow-rate on different microbial consortia was examined during anodic biofilm development, using inocula designed to enrich either aerobes/facultative species or anaerobes. All MFCs underperformed at high flow-rates in the early stages, however, the aerobic type - following anodic biofilm development - subsequently exhibited more marked improvement. Scanning electron microscopy showed some variation in biofilm formation where clumpy growth was associated with lower power. Over time both power and internal resistance increased for the low flow-rates perhaps explained by an evolving microflora that consequently changed redox potential. An overshoot was observed in power curves, which was attributed to increased internal resistance due to ionic depletion and/or microbial exhaustion. To the best of the authors' knowledge this is the first time that such phenomena are explained from the internal resistance perspective.


Consumer's Satisfaction with Community Pharmacies in Sindh, Pakistan.
The present research is the first comprehensive cross-sectional study of consumer satisfaction with community pharmacies in Sindh, Pakistan. Moreover, the study has also designed a nonorder theoretical model for consumer satisfaction with community pharmacies. This is a cross-sectional descriptive study from a general population of Sindh, Pakistan, with a total of four hundred and fifteen (n = 415) participants. A confirmatory factor analysis was used to verify the factor structure between Pharmaceutical services (PS), the Skill of Pharmacists, Non-pharmaceutical services (NPS), and pricing (P). Pearson correlation analysis, Kendall's tau correlation analysis, and Spearman's rho correlation analysis were used to identify the correlation between different factors, such as PS, SKP, NPS, and P. The 23-item scale that consisted of four elements have shown an acceptable root mean squared error of approximation (0.076), Cronbach's alpha (0.787), and Chi-square value (3.381) (P < 0.001). Of the respondents, 56.4% rated their satisfaction on pharmacist attitude, whereas 67.2%, 41.4%, and 51.8% were satisfied with other services, such as receipt provided on medication they take, prescription drug service and availability of pharmacies on the weekend and public holidays, respectively. This cross-sectional study confirms that there are relationships among PS, SPK, NPS, and P. Moreover, there is a lack of facilities in community pharmacies in Sindh, such as the unavailability of a consultation room, immunization services, information on routine health matters, and medication record.


Pyrrol[1,4]benzodiazepine antibiotics. Proposed structures and characteristics of the in vitro deoxyribonucleic acid adducts of anthramycin, tomaymycin, sibiromycin, and neothramycins A and B.
The pyrrol[1,4]benzodiazepine antibiotics anthramycin, tomaymycin, sibiromycin, and neothramycins A and B are potent antitumor agents that bind to DNA in a unique manner, resulting in some unusual biological consequences. This paper describes results on which the points of covalent linkage between the drugs (carbinolamine carbon atom) and DNA (N-2 of guanine) are deduced, as well as Corey-Pauling-Koltun (CPK) models for the various drug-DNA adducts. Predictions based upon these CPK models have been tested, and the results are reported in this paper. These tested experimental predictions include (1) instability of the drug-DNA adducts to denaturation of DNA, (2) saturation binding limits, (3) effect of drug binding on the structure of DNA, (4) lack of unwinding and in vitro strand breakage of closed-circular supercoiled simian virus 40 (SV-40) DNA, (5) sensitivity of the secondary structure of DNA to drug binding, (6) hydrodynamic properties of the drug-DNA adducts, (7) hydrogen bonding of the 9-phenolic proton in anthramycin to DNA, (8) structure-activity relationships, and (9) biological consequences of DNA damage, including cumulative damage and slow excision repair, double-strain breaks in DNA in repair-proficient cells, and the selective inhibition of H-strand DNA synthesis in mitochondria. The results are completely in accord with our postulated space-filling models.


Histone H3 modifications associated with differentiation and long-term culture of mesenchymal adipose stem cells.
Long-term culture of mesenchymal stem cells leads to a loss of differentiation capacity, the molecular mechanism of which remains not understood. We show here that expansion of adipose stem cells (ASCs) to late passage (replicative senescence) is associated with promoter-specific and global changes in epigenetic histone modifications. In undifferentiated ASCs, inactive adipogenic and myogenic promoters are enriched in a repressive combination of trimethylated H3K4 (H3K4m3) and H3K27m3 in the absence of H3K9m3, a heterochromatin mark. Sequential chromatin immunoprecipitation assays indicate that H3K4m3 and H3K27m3 co-occupy a fraction of nucleosomes on some but not all lineage-specific promoters examined. However in cultured primary keratinocytes, adipogenic and myogenic promoters are enriched in trimethylated H3K4, K27, and K9, illustrating two distinct epigenetic states of inactive promoters related to potential for activation. H3K4m3 and H3K27m3 stably mark promoters during long-term ASC culture indicating that loss of differentiation capacity is not due to alterations in these histone modifications on these loci. Adipogenic differentiation in early passage results in H3K27 demethylation and H3K9 acetylation specifically on adipogenic promoters. On induction of differentiation in late passage, however, transcriptional upregulation is impaired, H3K27 trimethylation is maintained and H3K9 acetylation is inhibited on promoters. In addition, the polycomb proteins Ezh2 and Bmi1 are targeted to promoters. This correlates with global cellular Ezh2 increase and H3K9 deacetylation. Promoter targeting by Ezh2 and Bmi1 in late passage ASCs suggests the establishment of a polycomb-mediated epigenetic program aiming at repressing transcription.


A selective CB2 agonist protects against the inflammatory response and joint destruction in collagen-induced arthritis mice.
Rheumatoid arthritis (RA) is a chronic, inflammatory, synovitis-dominated systemic disease with unknown etiology. RA is characterized by the involvement of multiple affected joints, symmetry, and invasive arthritis of the limbs, which can lead to joint deformity, cartilage destruction, and loss of function. Cannabinoid receptor 2 (CB2) has potent immunomodulatory and anti-inflammatory effects and is predominantly expressed in non-neuronal tissues. In the current study, the role of CB2 in the process of inflammatory bone erosion in RA was examined. The selective agonist or high-affinity ligand of CB2 (4-quinolone-3-carboxamides CB2 agonist, 4Q3C CB2 agonist, 4Q3C) significantly reduced the severity of arthritis, decreased histopathological findings, and markedly reduced bone erosion in collagen-induced arthritis (CIA) mice. In addition, 4Q3C prevented an increase in the nuclear factor-κB ligand (RANKL)/osteoprotegerin (OPG) ratio and inhibited the formation of osteoclasts in CIA mice. Furthermore, the expression of tumor necrosis factor-alpha, interleukin-1β, cyclooxygenase-2, and inducible nitric oxide synthase was lower in 4Q3C-treated CIA mice than in control CIA mice. Micro-computed tomography corroborated the finding that 4Q3C reduced joint destruction. These data clearly indicate that the CB2-selective agonist, 4Q3C, may have anti-inflammatory and anti-osteoclastogenesis effects in RA and may be considered to be a novel treatment for RA.


The RING finger protein SNURF modulates nuclear trafficking of the androgen receptor.
The androgen receptor (AR) is a transcription factor that mediates androgen action. We have used the green fluorescent protein (GFP) technique to investigate dynamics of nuclear trafficking of human AR in living cells. In the absence of ligand, the GFP-AR fusion protein is distributed between cytoplasm and nuclei. Androgen exposure leads to a rapid and complete import of GFP-AR to nuclei of CV-1 cells (>=90% nuclear in 30 minutes), whereas a pure antiandrogen, Casodex, elicits a slower (<40% nuclear in 30 minutes) and incomplete transfer. Unliganded ARs with mutations in the basic amino acids of the bipartite nuclear localization signal (NLS) within the second zinc finger and the hinge region are predominantly cytoplasmic and their androgen-dependent nuclear import is severely compromised ((3/4)20% nuclear in 30 minutes). Interestingly, substitutions of the Leu residues flanking the bipartite NLS lead to inefficient nuclear transfer in response to androgen ((3/4)20% nuclear in 30 minutes). The ligand-binding domain of AR, which represses bipartite NLS activity, contains an agonist-specific NLS. The small nuclear RING finger protein SNURF, which interacts with AR through a region overlapping with the bipartite NLS, facilitates AR import to nuclei and retards its export on hormone withdrawal. More AR is associated with the nuclear matrix in the presence than absence of coexpressed SNURF. We suggest that the SNURF-mediated tethering of AR in nuclei represents a novel mechanism for activating steroid receptor functions.


Error-prone PCR of a fungal xylanase for improvement of its alkaline and thermal stability.
Random mutagenesis was used to improve the alkaline and thermal stability of the xylanase (XynA) from Thermomyces lanuginosus. Error-prone PCR reactions were carried out; the PCR products were cloned into Escherichia coli and a library of 960 clones was selected on xylan-containing agar plates. The crude filtrates of positive xylanase producers were screened at 80 degrees C and tested separately at pH 10 for alkaline tolerance. The native XynA lost 80% activity after 90 min at 80 degrees C and lost 70% activity at pH 10. Conversely, the most thermostable variant, G41, retained 75% activity after 90 min at 80 degrees C and the best alkali-stable variant, G53, retained 93% activity at pH 10. Sequence analysis revealed four amino acid substitutions in G41 and a single substitution in G53. These variants, therefore, have improved thermal and alkaline stability and are ideal candidates for DNA shuffling experiments to produce a robust xylanase for industrial application.


The impact of United States law on medicine as a profession.
The evolution of US law has had an enormous influence on medicine as a profession, and much of this legal evolution can be attributed to changes in the science and practice of medicine. This changing legal framework and its intersection with medicine has many facets. Three of the most important facets concern the evolution of the no duty-to-treat principle and the role of modern health care financing and civil rights law in altering this rule, the manner in which advances in medicine led courts and legislatures to change the standards against which professional medical liability is measured, and the basic loss of highly preferential treatment under US laws aimed at preventing anticompetitive conduct by businesses. However, despite the impact on the profession of an evolving legal system, concern over the integrity of medical professionalism continues to significantly influence both laws and lawmakers, including legislatures, regulatory agencies, and the courts.


ERBB activation modulates sensitivity to MEK1/2 inhibition in a subset of driver-negative melanoma.
Melanomas are characterized by activating "driver" mutations in BRAF, NRAS, KIT, GNAQ, and GNA11. Resultant mitogen-activated protein kinase (MAPK) pathway signaling makes some melanomas susceptible to BRAF (BRAF V600 mutations), MEK1/2 (BRAF V600, L597, fusions; NRAS mutations), or other kinase inhibitors (KIT), respectively. Among driver-negative ("pan-negative") patients, an unexplained heterogeneity of response to MEK1/2 inhibitors has been observed. Analysis of 16 pan-negative melanoma cell lines revealed that 8 (50%; termed Class I) are sensitive to the MEK1/2 inhibitor, trametinib, similar to BRAF V600E melanomas. A second set (termed Class II) display reduced trametinib sensitivity, paradoxical activation of MEK1/2 and basal activation of ERBBs 1, 2, and 3 (4 lines, 25%). In 3 of these lines, PI3K/AKT and MAPK pathway signaling is abrogated using the ERBB inhibitor, afatinib, and proliferation is even further reduced upon the addition of trametinib. A potential mechanism of ERBB activation in Class II melanomas is minimal expression of the ERK1/2 phosphatase, DUSP4, as ectopic restoration of DUSP4 attenuated ERBB signaling through potential modulation of the ERBB ligand, amphiregulin (AREG). Consistent with these data, immunohistochemical analysis of patient melanomas revealed a trend towards lower overall DUSP4 expression in pan-negative versus BRAF- and NRAS-mutant tumors. This study is the first to demonstrate that differential ERBB activity in pan-negative melanoma may modulate sensitivity to clinically-available MEK1/2 inhibitors and provides rationale for the use of ERBB inhibitors, potentially in combination with MEK1/2 inhibitors, in subsets of this disease.


Physical activity and quality of life in severely obese individuals seeking bariatric surgery or lifestyle intervention.
Given that bariatric surgery (BS) and lifestyle intervention (LI) represent two vastly different approaches to treating severe obesity, there is growing interest in whether individuals who seek BS versus LI also differ on weight-related behaviors. In the present study, we compared BS- and LI-seekers on physical activity (PA) and sedentary behaviors (SB), and examined between-group differences in health-related quality of life (HRQoL), while controlling for PA. A sample of 34 LI-seekers were matched with 34 BS-seekers on gender, age, BMI, and PA monitor-daily wear time (age: 42.1±10.0 years; BMI: 45.6±6.5 kg/m2). PA and SB were assessed over a 7-day period via the SenseWear Armband (SWA). HRQoL was measured using the SF-36, with scores standardized to a population normal distribution (M=50, SD=10). Participants wore the SWA for 13.7±1.6 h/day. BS-seekers did not differ from LI-seekers on average min/d over the wear period spent in SB (641±117.1 vs. 638.4±133.4, p=0.62) or light (136.4±76.1 vs. 145.5±72.5, p=0.59) and moderate-to-vigorous (>1-min bouts=36.4±26.2 vs. 40.2±31.3, p=0.59; ≥10-min bouts=5.7±8.3 vs. 10.2±17.0, p=0.17) PA. BS-seekers reported significantly lower SF-36 physical functioning (42.4±10.9 vs. 49.0±6.8, p=0.004) and physical component summary (43.9±10.1 vs. 48.9±7.0) scores versus LI-seekers. BS-seeker group status was related to lower physical functioning (β=0.30, p=0.009), independent of gender, age, BMI, and daily PA. Findings suggest that seeking BS versus LI is not related to patterns of PA or SB, and that lower subjective physical functioning is not associated with lower overall PA levels in BS-seekers.


Backscatter-contour-attenuation joint estimation model for attenuation compensation in ultrasound imagery.
Ultrasound B-scan exhibits shadowing and enhancement artifacts due to acoustic wave propagation and spatially varying scatter attenuation across layers of tissues. These artifacts hide underlying echo signals that are truly clinically indicative of diseases. Attenuation compensation estimates and corrects for shadowing and enhancement artifacts, which improves the quality of ultrasound imaging. Block-based attenuation compensation methods, widely employed in commercial scanners, produce results with resolutions limited by the block size. To obtain higher spatial resolution (as desired for quantitative analysis), we present a backscatter-contour-attenuation (BCA) joint estimation model for attenuation compensation in pulse-echo imaging using a set of self-consistent partial differential equations and a contour evolution model. The problem is posed as reconstructing sources of information from observations. We derive the joint estimation model from minimizing a cost functional of separated attributes with region-based isotropic regularizations. A three-step alternating minimization method is adopted towards a tractable numerical solution. Detailed numerical methods are described. The efficacy of the proposed approach is demonstrated using simulated and real images.


Roxburgh was right: Aponogeton microphyllus and Aponogeton undulatus are distinct species.
Aponogeton microphyllus, previously placed under the synonymy of A. undulatus, is recognized here as a distinct species based on morphology, chromosome number, and molecular phylogenetics (nuclear ribosomal internal transcribed (ITS) spacer region). Observations on the type and live specimens revealed morphological differences between the two species. Aponogeton microphyllus flowered regularly and set seeds. Aponogeton undulatus flowered rarely, did not set seeds, but showed formation of young plantlets on the inflorescence axis. Similarly, different chromosome numbers were recorded in Aponogeton microphyllus and the two forms of A. undulatus, viz., AF1 and AF2, which occur in distinct populations. Aponogeton microphyllus exhibited polysomaty with root-tip cells showing 2n=40, 42, and 44 chromosomes. The two forms of A. undulatus, AF1 and AF2, showed 2n=84 and 86 chromosomes, respectively. Based on the ITS data, both species occupied two separate clades. Plastid trnK intron region indicated a close relationship between both species. Our study suggests the need for comprehensive phylogenetic analyses of A. undulatus across its distribution range based on more advanced techniques such as highthroughput sequencing data to understand the A. undulatus species complex and to detect natural hybrids of this species.


The Drosophila melanogaster PIF1 Helicase Promotes Survival During Replication Stress and Processive DNA Synthesis During Double-Strand Gap Repair.
PIF1 is a 5' to 3' DNA helicase that can unwind double-stranded DNA and disrupt nucleic acid-protein complexes. In Saccharomyces cerevisiae, Pif1 plays important roles in mitochondrial and nuclear genome maintenance, telomere length regulation, unwinding of G-quadruplex structures, and DNA synthesis during break-induced replication. Some, but not all, of these functions are shared with other eukaryotes. To gain insight into the evolutionarily conserved functions of PIF1, we created pif1 null mutants in Drosophila melanogaster and assessed their phenotypes throughout development. We found that pif1 mutant larvae exposed to high concentrations of hydroxyurea, but not other DNA damaging agents, experience reduced survival to adulthood. Embryos lacking PIF1 fail to segregate their chromosomes efficiently during early nuclear divisions, consistent with a defect in DNA replication. Furthermore, loss of the BRCA2 protein, which is required for stabilization of stalled replication forks in metazoans, causes synthetic lethality in third instar larvae lacking either PIF1 or the polymerase delta subunit POL32. Interestingly, pif1 mutants have a reduced ability to synthesize DNA during repair of a double-stranded gap, but only in the absence of POL32. Together, these results support a model in which Drosophila PIF1 functions with POL32 during times of replication stress but acts independently of POL32 to promote synthesis during double-strand gap repair.


The Importance of "Being There": a Qualitative Study of What Veterans with Depression Want in Social Support.
Social connectedness exerts strong influences on health, including major depression and suicide. A major component of social connectedness is having individual relationships with close supports, romantic partners, and other trusted members of one's social network. The objective of this study was to understand how individuals' relationships with close supports might be leveraged to improve outcomes for primary care patients with depression and at risk for suicide. In this qualitative study, we used a semi-structured interview guide to probe patient experiences, views, and preferences related to social support. We conducted interviews with 30 primary care patients at a Veterans Health Administration (VA) medical center who had symptoms of major depression and a close support. Thematic analysis of qualitative interview data examined close supports' impact on patients. We iteratively developed a codebook, used output from codes to sort data into themes, and selected quotations that exemplified themes for inclusion in this manuscript. "Being there" as an important quality of close supports emerged as a key concept. "Being there" was defined in three ways: physical proximity, frequent or responsive contact, or perceived availability. Close supports who were effective at "being there" possessed skills in intuitively sensing the patient's emotional state and communicating indirectly about depression. Three major barriers to involving close supports in depression care were concerns of overburdening the close support, a perception that awareness of the patient's depression would make the close support unnecessarily worried, and a desire and preference among patients to handle depression on their own. "Being there" represents a novel, patient-generated way to conceptualize and talk about social support. Suicide prevention initiatives such as population-level communication campaigns might be improved by incorporating language used by patients and addressing attitudinal barriers to allowing help and involvement of close supports.


Steroidogenic, Metabolic, and Immunological Markers in Dairy Cows Diagnosed With Cystic Ovarian Follicles at Early and Mid-Late Lactation.
The etiology of cystic ovarian follicles (COF) remains a conundrum with steroidogenic, immunological, and metabolic dysfunctions linked to its development. Studies suggest that COF development may occur as a result of disruption of the insulin signaling pathway and the severity of a negative energy balance in dairy cows, but mid to late lactation cows diagnosed with COF are unlikely to have issues with energy metabolism. Herein, we characterized the mRNA expression of steroidogenic (LHCGR, StAR, CYP11A1, 3β-HSD, CYP19A), immunological (IL-1β, IL-6, IL-8, TLR-4, TNF), and metabolic markers (IGF-1, IRS1) in follicular fluid; and plasma and follicular fluid levels of E2, IL-1β, glucose, and NEFA in early and mid-late lactation COF cows. Lactating dairy cows were diagnosed as having COF (n = 11, follicle >20 mm persistent for 7 days, absence of corpus luteum, and flaccid uterus) while 11 herdmates cycling with a dominant follicle were classified as the control cows. Cows diagnosed with COF were classified as early lactation (COF-E, n = 5) cows, <35 days in milk (DIM); or mid-late lactation (COF-M/L, n = 6), ≥118 DIM cows. Results revealed that mRNA expression StAR was greater (P < 0.01) in COF-E cows than COF-M/L cows and the control cows. The mRNA expression CYP19A1 was lower (P < 0.01) in COF-E cows and COF-M/L cows than in the control cows. The mRNA expression IL-6 and IRS-1 tended to be greater and lower, respectively, in COF-M/L cows compared to the control cows. The mRNA expression IGF-1 was greater (P < 0.01) in COF-E and COF-M/L cows than in the control cows. The plasma and follicular fluid concentration of NEFA was greater (P < 0.05) in COF-E cows than in COF-M/L and the control cows. Cows with COF-E had disturbances in steroidogenic and metabolic markers, while cows with COF-M/L had steroidogenic, immunological, and metabolic dysregulations, suggesting that COF pathogenesis may vary between early and mid-late lactation dairy cows.


Rotavirus persistence in cell cultures: selection of resistant cells in the presence of foetal calf serum.
The Lincoln strain of bovine rotavirus, cytocidal for bovine AU-BEK cells, can establish in the same cell cultures in the presence of foetal calf serum (FCS) a persistent infection that depends on selection of highly resistant cells. In fact, after the induction of the carrier state only a small fraction of the cell population was infected. The parental and the carried viruses are not demonstrably different, the cultures are resistant to superinfection by the homologous virus, the cultures can be cured by antiviral serum in the medium and uninfected resistant cell clones can be selected. The presence of FCS was essential during induction and maintenance of persistence. Its effects appear to be exerted not on the control of virus replication in fully sensitive cells but on the proliferation and selection of the resistant cells.