Physical mapping of the Fv-1 tropism host range determinant of BALB/c murine leukemia viruses.
The murine leukemia viruses (MuLVs) have different host ranges and were originally designated N-tropic and B-tropic if they replicated preferentially in vitro on NIH and BALB/c fibroblasts, respectively. It was later found that N-tropic MuLVs were in fact restricted in BALB/c cells, that B-tropic MuLVs were restricted in NIH cells, and that both viruses were restricted in (BALB X NIH) F1 cells. A single gene, Fv-1, with two alleles, Fv-1b and Fv-1n, determines this dominant restriction. A virus-encoded protein seems to carry the viral host range determinant which is recognized by the Fv-1 gene product. To map the viral DNA sequences encoding this determinant, we constructed viral DNA recombinants in vitro between the cloned infectious viral DNA genomes from BALB/c N-tropic and B-tropic MuLVs. Infectious recombinant MuLVs were recovered by microinjecting these recombinant DNAs into murine Fv-1- SC-1 cells and were subsequently tested in vitro for their host ranges (N- or B-tropic). We found that a short 302-base pair 5'-end fragment was necessary and sufficient to confer a specific host range to a recombinant. Our sequencing data revealed that this fragment codes for amino acid sequences in gag p30. They also showed that only two consecutive amino acid differences, Gln-ArgN- and Thr-GluB-, in p30 are responsible for the N- and B-tropic host ranges of the BALB/c MuLVs, respectively. Therefore, it appears that the Fv-1b and Fv-1n gene products can discriminate between these two p30 amino acid sequences.


Least Disturbed Condition for European Mediterranean rivers.
The present report describes a three-step approach that was used to characterize and define thresholds for the Least Disturbed Condition in Mediterranean streams of four different types, regarding organic pollution and nutrients, hydrological and morphological alterations, and land use. For this purpose, a common database composed of national reference sites (929 records) from seven countries, sampled for invertebrates, diatoms and macrophytes was used. The analyses of reference sites showed that small (catchment <100 km(2)) siliceous and non-siliceous streams were mainly affected by channelization, bank alteration and hydropeaking. Medium-sized siliceous rivers were the most affected by stressors: 25-43% of the samples showed at least slight alterations regarding channelization, connectivity, upstream dam influence, hydropeaking and degradation of riparian vegetation. Temporary streams were the least affected by hydromorphological changes, but they were nevertheless affected by alterations in riparian vegetation. There were no major differences between all permanent stream types regarding water quality, but temporary streams showed lower values for oxygenation (DO) and wider ranges for other variables, such as nitrates. A lower threshold value for DO (60%) was determined for this stream type and can be attributed to the streams' natural characteristics. For all other river types, common limits were found for the remaining variables (ammonium, nitrate, phosphate, total P, % of artificial areas, % of intensive and extensive agriculture, % of semi-natural areas in the catchment). These values were then used to select the list of reference sites. The biological communities were characterized, revealing the existence of nine groups of Mediterranean invertebrate communities, six for diatoms and five for macrophytes: each group was characterized by specific indicator taxa that highlighted the differences between groups.


Distribution of (1->4)-beta-galactans, arabinogalactan proteins, xylans and (1->3)-beta-glucans in tracheid cell walls of softwoods.
Polysaccharides were located in the walls of normal and compression wood tracheids of Pinus radiata (radiata pine), Picea sitchensis (Sitka spruce) and Picea abies (Norway spruce) by transmission electron microscopy using immunogold labelling with monoclonal antibodies to (1-->4)-beta-galactan (LM5), (1-->3)-beta-glucan, arabinogalactan proteins (AGPs) (MAC207) and heteroxylans (LM10 and LM11). In fully differentiated compression wood tracheids, (1-->4)-beta-galactan was found in the S2((L)) layer and, to a smaller extent, at the interface between the compound middle lamella and the S1 layer. (1-->4)-beta-Galactan appeared to be displaced from, or modified in, the S1 layer during cell wall formation. (1-->3)-beta-Glucan (callose) was confined to the helical cavities in the inner S2 layer of severe compression wood. MAC207 AGP glycan epitope was found exclusively in the S1 and S3 layers of normal wood tracheids and in the S1 and inner S2 layers of compression wood tracheids. Binding of LM10, which specifically recognizes unsubstituted or low-substituted xylans, occurred at similar locations to the MAC207 epitope, whereas binding of LM11, which recognizes more highly substituted as well as unsubstituted xylans, occurred throughout the tracheid walls with the exception of the primary wall. Immunogold labelling showed that the different wall layers of softwood tracheids have different polysaccharide compositions which change abruptly during cell wall formation.


Frequency of hyponatraemia and hypokalaemia in malnourished children with acute diarrhoea.
To determine the frequency of hyponatraemia and hypokalaemia in malnourished children with acute diarrhoea. This cross-sectional study was carried out at the Military Hospital, Rawalpindi, Pakistan, from September 2013 to March 2014, and comprised acute diarrhoea patients whose ages ranged from six months to five years. Blood samples for serum sodium and potassium were examined at the Armed Forces Institute of Pathology. Patients were labelled as having hyponatraemia, hypokalaemia, both or having normal serum sodium and potassium levels. Of the 80 patients, 49(61.3%) were boys and 31(38.7%) were girls with an overall mean age of 1.9±1.4 years. Besides, 41(51.3%) were aged below one year. The mean duration of diarrhoea was 3.2±1.7 days, with 53(66%) patients having the illness for 1-3 days. Hyponatraemia was observed in 26(32.5%) patients and hypokalaemia in 44(55%), whereas 10(12.5%) had no electrolyte imbalance. None of the participants had hypernatraemia or hyperkalaemia. Electrolyte disturbances among malnourished children may not be clinically evident, but diarrhoeal illness aggravated these imbalances.


Crackles in the early detection of asbestosis.
We studied 386 workers exposed to asbestos to assess the value of chest auscultation by a trained technician in detecting asbestosis as defined by previously reported clinical, physiologic, and roentgenologic criteria. The presence and degree of crackles were assessed at preselected basilar lung sites by a technician whose performance was validated by comparison with computer-generated time-expanded waveforms of tape recordings of lung sounds. Asbestosis was present in only 2.8% of the total population, but it was present in 8.6% of those with over 25 yr or more of employment. The technician correctly identified all the workers in whom the diagnosis was most certain, that is, those with all criteria positive. The overall true positive rate was 55%. The majority (94.8%) of those with no abnormal criteria were correctly classified. Auscultation by an objectively validated technician can be a useful noninvasive method for screening industrial populations exposed to asbestos.


Membrane depolarization induces protein kinase C translocation and voltage operated calcium channel opening in PU5-1.8 cells. Protein kinase C as a negative feedback modulator for calcium signalling.
Incubation of the murine macrophage tumour cell line PU5-1.8 in K+ (140 mM)-HEPES buffer induced depolarization of the membrane and the translocation of protein kinase C (PKC) to the subnuclear region. Membrane depolarization also induced an increase of intracellular free Ca2+ levels ([Ca2+]i) which was due to the Ca2+ influx. The amount of K(+)-mediated Ca2+ uptake was dependent on the Ca2+ concentration gradient as measured by indo-1 fluorescence and 45Ca2+ fluxes. The depolarization-mediated Ca2+ influx was suppressed by voltage sensitive Ca2+ channel blockers such as nifedipine and verapamil. Furthermore, in Na(+)-HEPES buffer, incubation of cells with a dihydropyridine agonist [3H]PN200-110 produced a dose-dependent saturable binding. On the other hand, short-term incubation of cells with phorbol 12-myristate 13-acetate (PMA) abolished the early phase of 45Ca2+ influx and the rise of indo-1 fluorescence. Depleting cells of PKC or incubating them with PKC inhibitors, H7 and sphingosine, enhanced the uptake of 45Ca2+ and the rise of indo-1 fluorescence. These observations suggest that membrane depolarization caused an activation of PKC and induced Ca2+ influx through the activation of dihydropyridine-sensitive, voltage-operated Ca2+ channels. Data also show that PKC may act as a negative modulator in controlling the Ca2+ response by closing the voltage-operated Ca2+ channel and/or by enhancing the Ca(2+)-ATPase activity during membrane depolarization in PU5-1.8 cells.


Homology modeling of the structure of bacterial acetohydroxy acid synthase and examination of the active site by site-directed mutagenesis.
Acetohydroxy acid synthase (AHAS, EC 188.8.131.52) catalyzes the thiamin pyrophosphate (TPP)-dependent decarboxylation of pyruvate and condensation of the resulting two-carbon moiety with a second alpha-keto acid. It belongs to a family of homologous, TPP-dependent enzymes which catalyze different reactions which start from decarboxylation of alpha-keto acids. A model for the structure of Escherichia coli AHAS isozyme II, based on its homology with pyruvate oxidase and experimental testing of the model by site-directed mutagenesis, has been used here to study how AHAS controls the chemical fate of a decarboxylated keto acid. Because of the potential conformational freedom of the reacting substrates, residues interacting with the substrate could not be identified directly from the model of AHAS. Three residues were considered as candidates for involvement in the recognition of alpha-ketobutyrate, as the amino acids at these sites in a unique low-specificity AHAS are different from those in typical AHASs, which are highly specific for reaction with alpha-ketobutyrate as second substrate, in preference to pyruvate. These residues were altered in AHAS II by site-directed mutagenesis. Replacement of Trp464 lowers the specificity by at least 1 order of magnitude, with minor effects on the activity or stability of the enzyme, suggesting that Trp464 contributes > or = 1.3 kcal mol-1 to interaction with the "extra" methyl of alpha-ketobutyrate. Mutations of Met460 or Thr70 have small effects on specificity and do affect other properties of the protein. A model for enzyme-substrate interactions can be proposed on the basis of these results. The model of AHAS also explains previously reported spontaneous mutants of AHAS resistant to sulfonylurea herbicides, which probably bind in the narrow depression which provides access to the bound TPP. A role for the C terminus of the enzyme polypeptide in determination on the reaction pathway is also possible.


Relation between patterns of intrinsic network connectivity, cognitive functioning, and symptom presentation in trauma-exposed patients with major depressive disorder.
The present study investigated resting fMRI connectivity within the default mode (DMN), salience (SN), and central executive (CEN) networks in relation to neurocognitive performance and symptom severity in trauma-exposed patients with major depressive disorder (MDD). Group independent component analysis was conducted among patients with MDD (n = 21), examining DMN, SN, and CEN connectivity in relation to neurocognitive performance and symptom severity. Activation in these networks was also compared between the patient group and healthy controls (n = 20). Among the patient group, higher levels of performance on measures of verbal memory and executive functioning were related to increased connectivity within the DMN (i.e., inferior parietal lobe; precuneus). Greater depression severity was related to reduced connectivity between the SN and a node of the DMN (i.e., posterior cingulate cortex) and higher depersonalization symptoms were related to enhanced connectivity between the SN and a node of the DMN (i.e., middle temporal gyrus). Higher symptoms of depersonalization were also associated with reduced integration of the DMN with the medial frontal gyrus. Relative to controls, patients with MDD showed greater connectivity of the ventromedial prefrontal cortex within the DMN. Intrinsic connectivity network patterns are related to cognitive performance and symptom presentation among trauma-exposed patients with MDD.


Why do some participants in colorectal cancer screening choose not to undergo colonoscopy following a positive test result? A qualitative study.
Our aim was to investigate why participants opted out of colonoscopy following a positive screening result for colorectal cancer. Semi-structured, qualitative, single interviews. We audio-recorded and transcribed all interviews verbatim and used Strauss and Corbin's concept of open, axial, and selective coding to identify the main categories shared across all interviews. These formed the basis of our findings. A Danish national colorectal cancer screening programme. Single interviews with 13 participants who declined to have a colonoscopy. Reasons to decline colonoscopy after positive screening test. Participants gave 42 different reasons for deciding not to have a colonoscopy and we coded them into nine main categories; Practical barriers, Discomfort of the examination, Personal integrity, Multimorbidity, Feeling healthy, Not having the energy, Belief that cancer is not present, Risk of complications, and Distrust in the accuracy of the iFOBT. Our findings suggest that some practical barriers could be quite easily addressed, by offering the participants alternative management and procdures. Further research is needed to examine how widely our findings are represented in the general population, and how general practitioners should consult with patients who have opted out of colonoscopy, despite a positive screening result. Key points Some screening participants are reluctant to proceed with further diagnostic tests for colorectal cancer following a positive screening result. • Interviews with people, who had refused a follow-up colonoscopy, discovered nine categories (42 reasons) of reasons for refusal. • Reluctance can be addressed by offering support with pre-procedure preparations and alternatives to colonoscopy. • General practitioners face ethical dilemmas and challenges, when patients at risk of colorectal cancer decline to proceed with screening.


Targeting Mitophagy in Alzheimer's Disease.
Mitochondria perform many essential cellular functions including energy production, calcium homeostasis, transduction of metabolic and stress signals, and mediating cell survival and death. Maintaining viable populations of mitochondria is therefore critical for normal cell function. The selective disposal of damaged mitochondria, by a pathway known as mitophagy, plays a key role in preserving mitochondrial integrity and quality. Mitophagy reduces the formation of reactive oxygen species and is considered as a protective cellular process. Mitochondrial dysfunction and deficits of mitophagy have important roles in aging and especially in neurodegenerative disorders such as Alzheimer's disease (AD). Targeting mitophagy pathways has been suggested to have potential therapeutic effects against AD. In this review, we aim to briefly discuss the emerging concepts on mitophagy, molecular regulation of the mitophagy process, current mitophagy detection methods, and mitophagy dysfunction in AD. Finally, we will also briefly examine the stimulation of mitophagy as an approach for attenuating neurodegeneration in AD.


Effect of skipping breakfast on cardiovascular risk factors: a grade-assessed systematic review and meta-analysis of randomized controlled trials and prospective cohort studies.
Skipping breakfast is one of the most prevalent irregular eating habits. Several pieces of evidence have reported the association between breakfast omission and a higher risk of cardiovascular diseases. Numerous publications have focused on the impact of skipping breakfast on various cardiovascular risk factors. Therefore, the current systematic review and meta-analysis aimed to assess this impact, especially with regard to anthropometric measurements, serum lipid profiles, blood pressure, and glycemic control indicators. A comprehensive search was performed in PubMed, Web of Science, Embase, Scopus, and the Cochrane Central Register of Controlled Trials up to 1 April 2023. A total of 11 eligible trials were identified to evaluate the combined effects of skipping breakfast. Final integrated results demonstrated that breakfast omission significantly decreased the body weight (mean difference = -0.66, 95% CI: -1.09 to -0.24, p = 0.002, I2 = 0.0) and increased the level of serum low-density lipoprotein cholesterol (LDL-C) (mean difference = 9.89, 95% CI: 5.14 to 14.63, p = 0.000, I2 = 17.3). Subgroup analysis also revealed potential factors that may affect the outcomes, for example, the physiological condition of participants, duration, gender, and type of breakfast. In conclusion, skipping breakfast may reduce body weight while increasing the level of serum LDL-C at the same time. In view of the limited trials, further studies are needed to expound the role of breakfast omission in cardiovascular diseases.


A Prospective Multicentric Postmarketing Observational Study to Characterize the Patient Population with Reduced Gastrointestinal Motility among Indian Diabetic Patients Receiving Itopride: The Progress Study.
This study was intended to assess the clinical profile of Indian diabetic patients with reduced gastrointestinal (GI) motility and to understand the role of itopride in addressing reduced GI motility (gastroparesis) symptoms and maintaining glycemic control. Patients with established reduced GI motility (scintigraphy), with varying degree of GI symptoms, receiving itopride 150 mg as per physicians' discretion were enrolled. Clinical profile, changes in symptom severity, glycemic indices, tolerability, and quality of life (QoL) after 8-week therapy (Patient assessment of upper GI disorders-QoL [PAGI-QoL]) were assessed. Mean ± standard deviation age of enrolled population (n = 41) was 51.8 ± 12.39 years. Average duration of gastroparesis since underlying etiology was 67.7 ± 59.76 months. Common symptoms reported at baseline were bloating (68.3%), postprandial fullness (61.0%), nausea (51.2%), early satiety (41.5%), heartburn (39.0%), and vomiting (9.8%). Itopride therapy resulted in significant improvement in all symptoms (P < 0.001), which correlated with improved QoL (PAGI-QoL score reduction: 13.8 ± 11.48; P < 0.0001). Moreover, significant improvement in glycemic indicators was also evident (mean change from baseline hemoglobinA1c -0.5 ± 1.18; fasting plasma glucose -15.3 ± 43.61; postprandial plasma glucose -24.6 ± 57.20). Itopride showed effectiveness in addressing symptoms of reduced GI motility in diabetics, with improved QoL. Significant improvement in glycemic indices was also evident posttreatment with itopride. This study sheds light on the role of prokinetics, not only for symptom relief but also for improving glycemic control in diabetic patients with reduced GI motility, thus providing a holistic approach for the management of these patients.


Living on the Edge: Settlement Patterns by the Symbiotic Barnacle Xenobalanus globicipitis on Small Cetaceans.
The highly specialized coronulid barnacle Xenobalanus globicipitis attaches exclusively on cetaceans worldwide, but little is known about the factors that drive the microhabitat patterns on its hosts. We investigate this issue based on data on occurrence, abundance, distribution, orientation, and size of X. globicipitis collected from 242 striped dolphins (Stenella coeruleoalba) that were stranded along the Mediterranean coast of Spain. Barnacles exclusively infested the fins, particularly along the trailing edge. Occurrence, abundance, and density of X. globicipitis were significantly higher, and barnacles were significantly larger, on the caudal fin than on the flippers and dorsal fin. Barnacles were found more frequently and in greater numbers on the dorsal rather than ventral side of the caudal fin and on the central third of dorsal and ventral fluke surfaces. Nearly all examined individuals attached with their cirral fan oriented opposite to the fluke edge. We suggest that X. globicipitis may chemically recognize dolphins as a substratum, but fins, particularly the flukes, are passively selected because of creation of vortices that increase contact of cyprids with skin and early survival of these larvae at the corresponding sites. Cyprids could actively select the trailing edge and orient with the cirri facing the main direction of flow. Attachment on the dorsal side of the flukes is likely associated with asymmetrical oscillation of the caudal fin, and the main presence on the central segment of the flukes could be related to suitable water flow conditions generated by fluke performance for both settlement and nutrient filtration.


Sensitive trapping and gas chromatographic method for vinyl chloride in air samples.
A simple and sensitive method is described for determining vinyl chloride monomer in air. Vinyl chloride monomer from a sample of air is trapped onto a Tenax GC trap cooled with Dry Ice. The trap is installed on a gas chromatograph by a simple valve arrangement. The trap is heated, and the entire sample is flushed onto the gas chromatographic column. Vinyl chloride is determined at 6 and 60 ppb (v/v) levels with recoveries ranging from 79 to 104%.


[Hiatal hernia of the esophagus and GERD as a cause of hemorrhage].
Bleeding complications arise in 1/4 of patients with hiatal hernia and GERD, and are the cause in 10% of all acute and 1/3 of chronic foregut bleedings. Most common bleeding disorders directly related to hiatal hernia and GERD are: hiatal hernia ulcers, erosive esophagitis, esophageal ulcers, peptic strictures and Barrett esophagus. The aim of this review article is to point out a significance of proper diagnosis and treatment for conditions bonded with hiatal hernia and GERD which can lead to severe esophageal bleedings. Detailed etiology, incidence, diagnostic algorithm and treatment of Cameron lesions, prolapse gastropathy, erosive esophagitis, peptic esophageal ulcers and postoperative complications related to hiatal hernia and GERD are presented in this article.


Zinc-65 in marine organisms along the Oregon and Washington coasts.
The concentration of zinc-65 in marine animals and plants near the mouth of the Columbia River is presented. Amounts of radiozinc found in the biota diminished rapidly with the distance from the river mouth. The highest levels were found in plankton, algae, and mollusks. Of the human foods, oysters exhibited the highest levels.


n-3 Fatty Acid Supplementation and Leukocyte Telomere Length in Patients with Chronic Kidney Disease.
DNA telomere shortening associates with the age-related increase cardiovascular disease (CVD) risk. Reducing oxidative stress, could modify telomere erosion during cell replication, and CVD risk in patients with chronic kidney disease (CKD). The effect of n-3 fatty acids and coenzyme Q10 (CoQ) on telomere length was studied in a double-blind placebo-controlled trial in CKD. Eighty-five CKD patients were randomized to: n-3 fatty acids (4 g); CoQ (200 mg); both supplements; or control (4 g olive oil), daily for 8 weeks. Telomere length was measured in neutrophils and peripheral blood mononuclear cells (PBMC) at baseline and 8 weeks, with and without correction for cell counts. Main and interactive effects of n-3 fatty acids and CoQ on telomere length were assessed adjusting for baseline values. F₂-isoprostanes were measured as markers of oxidative stress. There was no effect of n-3 fatty acids or CoQ on neutrophil or PBMC telomere length. However, telomere length corrected for neutrophil count was increased after n-3 fatty acids (p = 0.015). Post-intervention plasma F₂-isoprostanes were negative predictors of post-intervention telomere length corrected for neutrophil count (p = 0.025).The effect of n-3 fatty acids to increased telomere length corrected for neutrophil count may relate to reduced oxidative stress and increased clearance of neutrophils with shorter telomeres from the circulation. This may be a novel mechanism of modifying CVD risk in CKD patients.


Combination of serum carcino-embryonic antigen and C-reactive protein--a useful test in preoperative staging of colorectal cancer.
The value of the simultaneous estimation of serum carcino-embryonic antigen (CEA) and C-reactive protein (CRP) concentrations in staging patients with colorectal neoplasias was evaluated. The study included 95 patients with benign or malignant epithelial tumors of colon and rectum. The simultaneous negative values of CEA and CRP were able to exclude stage D tumors with a specificity of 92.6% (P less than 0.001). Simultaneously positive values of CEA and CRP were able to diagnose stage C or D tumors with a specificity of 92.1% (P less than 0.001). It is concluded that the combination of CEA and CRP tests can substantially contribute to the preoperative staging and assessment of the extent of colorectal cancer.


The influence of the N-terminal region proximal to the core domain on the assembly and chaperone activity of αB-crystallin.
αB-Crystallin (HSPB5) is a small heat-shock protein that is composed of dimers that then assemble into a polydisperse ensemble of oligomers. Oligomerisation is mediated by heterologous interactions between the C-terminal tail of one dimer and the core "α-crystallin" domain of another and stabilised by interactions made by the N-terminal region. Comparatively little is known about the latter contribution, but previous studies have suggested that residues in the region 54-60 form contacts that stabilise the assembly. We have generated mutations in this region (P58A, S59A, S59K, R56S/S59R and an inversion of residues 54-60) to examine their impact on oligomerisation and chaperone activity in vitro. By using native mass spectrometry, we found that all the αB-crystallin mutants were assembly competent, populating similar oligomeric distributions to wild-type, ranging from 16-mers to 30-mers. However, circular dichroism spectroscopy, intrinsic tryptophan and bis-ANS fluorescence studies demonstrated that the secondary structure differs to wild type, the 54-60 inversion mutation having the greatest impact. All the mutants exhibited a dramatic decrease in exposed hydrophobicity. We also found that the mutants in general were equally active as the wild-type protein in inhibiting the amorphous aggregation of insulin and seeded amyloid fibrillation of α-synuclein in vitro, except for the 54-60 inversion mutant, which was significantly less effective at inhibiting insulin aggregation. Our data indicate that alterations in the part of the N-terminal region proximal to the core domain do not drastically affect the oligomerisation of αB-crystallin, reinforcing the robustness of αB-crystallin in functioning as a molecular chaperone.


Enhanced mutagenesis of UV-irradiated simian virus 40 occurs in mitomycin C-treated host cells only at a low multiplicity of infection.
Treatment of monkey kidney cells with mitomycin C (MMC) 24 h prior to infection with UV-irradiated simian virus 40 (SV40) enhanced both virus survival and virus mutagenesis. The use of SV40 as a biological probe has been taken as an easy method to analyse SOS response of mammalian cells to the stress caused by DNA damage or inhibition of DNA replication. The mutation assay we used was based on the reversion from a temperature-sensitive phenotype (tsA58 mutant) to a wild-type phenotype. The optimal conditions for producing enhanced survival and mutagenesis in the virus progeny were determined with regard to the multiplicity of infection (MOI). Results showed that the level of enhanced mutagenesis observed for UV-irradiated virus grown in MMC-treated cells was an inverse function of the MOI, while enhanced survival was observed at nearly the same level regardless of the MOI. For the unirradiated virus, almost no increase in the mutation of virus progeny issued from MMC-treated cells was observed, while a small amount of enhanced virus survival was obtained. These results show that enhanced virus mutagenesis and enhanced virus survival can be dissociated under some experimental conditions. Enhanced virus mutagenesis, analogous to the error-prone replication of phages in SOS-induced bacteria, was observed, at least for SV40, only when DNA of both virus and host cells was damaged and when infection occurred with a small number of viral particles. We therefore hypothesize that an error-prone replication mode of UV-damaged templates is observed in induced monkey kidney cells.


Comparison of the Health Utilities Index Mark 3 (HUI3) and the EuroQol EQ-5D in patients treated for intermittent claudication.
The Health Utilities Index Mark 3 (HUI3) and the EuroQol EQ-5D (EQ-5D) were compared to each other and to other quality-of-life (QoL) measures in patients treated for intermittent claudication. A total of 88 patients with intermittent claudication completed the HUI3, EQ-5D, RAND 36-Item Health Survey 1.0, time tradeoff, standard gamble, and rating scale before revascularization and at follow-up at 1 month, 3 months, and 1 year. The effect of treatment on the HUI3 and EQ-5D dimensions and the overall scores, calculated using published formulas based on societal preferences, were compared. After 1 month of treatment, the majority of patients showed improvement on the HUI3 dimensions ambulation and pain and on the EQ-5D dimensions mobility, usual activities, and pain/discomfort. The mean HUI3 score was significantly higher than the mean EQ-5D score (0.66 and 0.57, respectively, p < 0.01) before treatment. After treatment, however, they were not significantly different from each other (e.g., 12 months after treatment: 0.77 and 0.75, respectively (p > 0.05). After 1 month, the scores did not change significantly over time (p > 0.05). The intraclass correlation coefficient between changes over time in the HUI3 and EQ-5D scores was 0.30, with other health-related quality-of-life (HRQoL) measures the correlations for HUI3 and EQ-5D were very similar. In conclusion, both the HUI3 and EQ-5D demonstrated an effect of treatment in patients with intermittent claudication; in addition, they showed similar relationships with other (HRQoL) measures. To demonstrate the effect of revascularization in patients with intermittent claudication, however, clinicians and researchers should be aware of the differences in the mean HUI3 and EQ-5D scores.


TNFR-Associated Factors 2 and 5 Differentially Regulate the Instructive IL-6 Receptor Signaling Required for Th17 Development.
IL-17-producing CD4(+) T cells (Th17 cells) regulate host defense and immune pathogenesis, and IL-6 plays an important role for the differentiation of Th17 cells. We have previously identified that TNFR-associated factor (TRAF)5 binds to the signal-transducing receptor gp130 through the C-terminal TRAF domain and inhibits Th17 development mediated by IL-6. Although gp130 has TRAF-binding motifs that can be recognized by other TRAF family proteins, it is unclear how TRAFs regulate IL-6-driven Th17 differentiation in general. Using retrovirus-mediated gene complementation and gene silencing approaches, we found that not only TRAF5 but also TRAF2 restrained the IL-6R signaling, whereas TRAF1, TRAF3, TRAF4, and TRAF6 did not. Traf2 silencing further promoted the ability of naive CD4(+) T cells from Traf5(-/-) mice to differentiate into Th17 cells. Notably, TRAF5 but not TRAF2 expressed in naive CD4(+) T cells was rapidly downregulated after TCR triggering, which indicates that TRAF5 specifically inhibits instructive IL-6 signals in the initial stage of Th17 development. Collectively, our results demonstrate a dedicated role for TRAF2 and TRAF5 in the process of IL-6-mediated Th17 development and a differential role for TCR signaling in regulation of TRAF2 and TRAF5. Therefore, both TRAF2 and TRAF5 work as important regulators of the IL-6R signaling needed for Th17 development.


The perception of stress and its impact on health in poor communities.
With the increased understanding of the relationship between stress and disease, the role of stress in explaining persistent disparities in health outcomes has received growing attention. One body of research has focused on allostatic load--the "wear and tear" that results from chronic or excessive activation of the stress response. Other research has looked at the link between stress and health behaviors. In this study, we conducted 7 focus groups with a total of 56 people to understand how people living in Highbridge, South Bronx, New York, a low income community with poor health outcomes, perceive stress and its relationship to health. Focus group participants described a direct causal pathway between stress and poor health as well as an indirect pathway through health behaviors, including uncontrolled eating, sleep deprivation, substance abuse, smoking, violence and aggression, and withdrawal and inactivity. Participants articulated a number of theories about why stress leads to these unhealthy behaviors, including self-medication, adaptive behavior, discounting the future, depletion of willpower, and competing priorities. Their nuanced understanding of the link between stress and health elucidates the mechanisms and pathways by which stress may result in disparities in health outcomes and create challenges in changing health behaviors in poor communities like the South Bronx.


Distribution of High-Risk Human Papillomavirus Genotypes and Multiple Infections in Preneoplastic and Neoplastic Cervical Lesions of Unvaccinated Women: A Cross-sectional Study.
The aim of the study was to investigate the distribution of high-risk (HR) human papillomavirus (HPV) genotypes and the role of multiple infection in preneoplastic and neoplastic cervical lesions, according to histology, age, and the number of genotypes per infection. Nine hundred eighty-eight women affected by known HPV-related cervical lesions and attending the European Institute of Oncology, Milan, Italy, from December 2006 to December 2014, were selected for a cross-sectional study. Prevalence of HPV genotypes was calculated by histology and the number of genotypes per infection. Univariate and multivariable cervical intraepithelial neoplasia (CIN) 2-3 versus CIN 1 risks were estimated by logistic regression models. Overall, HPV 16 (53.1%), HPV 31 (15.1%), and HPV 58 (6.4%) were the most frequent genotypes in precancerous lesions. At multivariable analysis, HPV 16 (p = .02), 18 (p = .013), and 56 (p = .01) were significantly associated to worsen histology, whereas HPV 39 (p = .03) and 45 (p = .03) were statistically correlated only to the increasing number of genotypes per infections. Human papillomavirus 33 was the only genotype significantly related to both the number of genotypes per infection (p = .005) and age (p = .03). Infections by HR-HPV (odds ratio [OR] = 9.48, 95% CI = 3.77-23.8, p < .001), HPV genotypes covered by current vaccines (OR = 6.28, 95% CI = 4.05-9.75, p < .001), single HPV genotype (OR = 8.13, 95% CI = 4.12-16.0, p < .001), as well as age (OR = 1.13, 95% CI = 1.07-1.19, p < .001) were significantly associated to higher risk of CIN 2-3. The most of CIN 2+ lesions are sustained by HR-HPV genotypes, especially the ones covered by 9-valent vaccine; therefore, the widespread use of prophylactic HPV vaccines could significantly reduce the incidence of preneoplastic and neoplastic cervical lesions.


The application of future technologies to medical informatics.
Physician: "Condyloma, Toxoplasmosis, Blepharoplasty, and Fibroadenoma." Technoguru: "Pardon?" Physician (referring to "PCDR, Physician's Computer Desk Reference): "Carrier Sense Multiple Access, Spread Spectrum, Application Programming Interface, and Clustered Indexes." Technoguru: "Oh, now you're talking! How many do you want?" Until such time as computer scientists holding degrees in medicine become de rigueur, there will inevitably be conversations such as these. A pediatrician friend once told me that he could teach me in 30 days what I would need to know to handle 95 percent of the cases he sees. To handle the other 5 percent would still require 8 years of postgraduate medical education. The corollary for the application of technology is that I can teach you how to use a personal computer, and even to do a little programming, but to build a robust, mission-critical system for a production health care environment, well, back to school you go.


Transfer of antirotaviral antibodies from mothers to their infants.
Levels of rotavirus-specific immunoglobulin G (IgG), IgA, IgM, and secretory immunoglobulin in maternal and cord serum, colostrum and milk, and infants' stools were measured by enzyme-linked immunosorbent assay in 92 mothers and their infants. Although antirotaviral IgG, IgA, and secretory immunoglobulin were present in most maternal sera, only IgG crossed the placenta. All samples of colostrum and milk tested contained antirotaviral secretory immunoglobulin and IgA except those of two women in whom IgA deficiency was subsequently described. Specific IgM and IgG were also detected in many colostral samples. Antirotaviral IgA was detected in many colostral samples. Antirotaviral IgA was detected in stools of breast-fed but not bottle-fed neonates. Apparently the human infant receives rotaviral antibodies both transplacentally and via maternal colostrum and milk.


Young children do not require perceptual-motor feedback to solve Aesop's Fable tasks.
Aesop's Fable tasks-in which subjects drop objects into a water-filled tube to raise the water level and obtain out-of-reach floating rewards -have been used to test for causal understanding of water displacement in both young children and non-human animals. However, a number of alternative explanations for success on these tasks have yet to be ruled out. One hypothesis is that subjects may respond to perceptual-motor feedback: repeating those actions that bring the reward incrementally closer. Here, we devised a novel, forced-choice version of the Aesop's Fable task to assess whether subjects can solve water displacement tasks when this type of feedback is removed. Subjects had to select only one set of objects, or one type of tube, into which all objects were dropped at once, and the effect the objects had on the water level was visually concealed. In the current experiment, fifty-five 5-9 year old children were tested in six different conditions in which we either varied object properties (floating vs. sinking, hollow vs. solid, large vs. small and too large vs. small objects), the water level (high vs. low) and/or the tube size (narrow vs. wide). We found that children aged 8-9 years old were able to solve most of the water displacement tasks on their first trial, without any opportunity for feedback, suggesting that they mentally simulated the results of their actions before making a choice. Children aged 5-7 years solved two conditions on their first trial (large vs. small objects and high- vs. low-water levels), and learnt to solve most of the remaining conditions over five trials. The developmental pattern shown here is comparable to previous studies using the standard Aesop's Fable task, where eight year olds are typically successful from their first trial and 5-7 year olds learn to pass over five trials. Thus, our results indicate that children do not depend on perceptual-motor feedback to solve these water displacement tasks. The forced-choice paradigm we describe could be used comparatively to test whether or not non-human animals require visual feedback to solve water displacement tasks.


Ribozyme inhibition of alphavirus replication.
A model system to examine the expression and antiviral activity of trans-acting ribozymes in mammalian cells has been developed and evaluated. Hairpin ribozymes were engineered to cleave a specific site, identified by a combinatorial activity-based selection method, within genomic and subgenomic RNA species of Sindbis virus. Transiently transfected cells expressed moderate levels of ribozyme (approximately 50,000 molecules/cell) with predominant nuclear localization and a short half-life (23 min). Stable cell lines expressed ribozymes at modest levels (approximately 2,000 molecules/cell). Ribozyme-mediated RNA cleavage activity was detected in cell extracts. Clonal cell lines were challenged with recombinant Sindbis virus, and viral replication was examined using plaque formation and green fluorescent protein assays. Significant inhibition of viral replication was observed in cells expressing the active antiviral ribozyme, and lower levels of inhibition in control cells expressing inactive or irrelevant ribozymes. These findings are consistent with a model in which inhibition of viral replication occurs via ribozyme cleavage of viral RNAs, suggesting that ribozymes may represent useful antiviral agents.


Sound localization in patients with unilateral cochlear implants.
To evaluate sound localization ability in totally deaf patients with unilateral cochlear implants and to estimate the ability to improve this function by training. A controlled case series. Nine patients with monaural cochlear implants were asked to identify the source of 50 randomly distributed sound stimuli coming from five different directions anteriorly or laterally. After some training, patients were retested. There were two control groups. One comprised nine adults with normal hearing, and the other comprised nine adults with normal hearing but one ear plugged. The mean initial score of the study group (maximal score 100) was 41.5 (range 23-63). Patients who had used cochlear implants longer had better sound localization ability (Spearman's correlation coefficient = 0.92). On retesting, after an average of 6.3 training sessions, their mean score improved to 66 (range 32-95), (p = 0.008). The improvement was more pronounced in postlinguals than in prelinguals (p = 0.016). Spontaneous development of sound localization ability in totally deaf patients with unilateral cochlear implants is proportional to the time interval between implantation and initial testing. Improvement appears to be influenced by training, and to be greater in postlingual than in prelingual implantees.


Drosophila as a Robust Model System for Assessing Autophagy: A Review.
Autophagy is the process through which a body breaks down and recycles its own cellular components, primarily inside lysosomes. It is a cellular response to starvation and stress, which plays decisive roles in various biological processes such as senescence, apoptosis, carcinoma, and immune response. Autophagy, which was first discovered as a survival mechanism during starvation in yeast, is now known to serve a wide range of functions in more advanced organisms. It plays a vital role in how cells respond to stress, starvation, and infection. While research on yeast has led to the identification of many key components of the autophagy process, more research into autophagy in more complex systems is still warranted. This review article focuses on the use of the fruit fly Drosophila melanogaster as a robust testing model in further research on autophagy. Drosophila provides an ideal environment for exploring autophagy in a living organism during its development. Additionally, Drosophila is a well-suited compact tool for genetic analysis in that it serves as an intermediate between yeast and mammals because evolution conserved the molecular machinery required for autophagy in this species. Experimental tractability of host-pathogen interactions in Drosophila also affords great convenience in modeling human diseases on analogous structures and tissues.


Inhibition of human blood clotting by extracts of Ascaris suum.
The effects of soluble extracts of Ascaris suum on human blood coagulation were investigated. Whole worm supernatant solution prolonged the whole blood clotting time and the kaolin-activated, partial thromboplastin time but it did not alter the prothrombin time. These data indicate impairment of the intrinsic pathway of blood coagulation. Whole worm supernate inhibited platelet aggregation induced by ADP and ristocetin. This supernate did not affect fibrinolysis. The maximal concentration of anticoagulant activity was found in the pseudocoelomic fluid of the worm. Activity was also noted in the cuticle and secretory/excretory products. Perhaps inhibition of blood clotting by helminths may facilitate their passage through the blood stream.


[Central administration of the amyloid beta-peptide (25-35) and individual features of cognitive behavior of rats].
Individual features of cognitive behavior of rats induced by the administration of the aggregated amyloid beta3-peptide (25-35) (Abeta25-35) into the lateral ventricles of the brain were studied. A new approach to the estimation of individual differences in the effects of Abeta25-35 was applied. Navigation training was performed in one short session with a random position of a target; sessions were repeated with the intervals of a few days. It was found for the first time that, at the early stage of its action, Abeta25-35 did not disturb the cognitive behavior in 50% of rats. The activity of the antioxidant enzymes superoxide dismutase and catalase in the cortex and the hippocampus of animals both with and without disturbance of cognitive behavior was significantly decreased. We suggest that the differences in individual responses may be due to phenotypic features of the animals. At the early stage of its action, the aggregated Abeta may induce a compensatory response preventing the disturbance of cognitive processes.


A simulated annealing approach to the search problem of protein crystallography.
With the growing availability of computer power it has become routine to perform exhaustive multidimensional searches in protein crystallography. Specifically, in cases where homologous or partially homologous structures are available, the initial interpretation of poor electron density maps is done by performing computer-intensive rotational and translational searches in real space. Often such calculations of the best fit between structure and map cannot even be attempted owing to the vast computing effort involved (years of MicroVax II time). Here, the combinatorial optimization method, simulated annealing, is shown to reduce substantially the computing effort involved and also to permit computations that are beyond the reach of current algorithms. This is illustrated with practical examples involving the structure determinations of the human histocompatibility antigen HLA-A2 and an influenza virus hemagglutinin-sialic acid complex.


A new ceramic interspinous process spacer for lumbar spinal canal stenosis.
Because surgery in elderly patients should be minimally invasive, interspinous process distraction has been widely used in this group to treat lumbar canal stenosis. We developed a new interspinous process distraction spacer composed of hydroxyapatite ceramic. In this work, we demonstrate the usefulness of this novel device. Since 2003, we operated on 19 elderly patients with lumbar canal stenosis, including 14 men and five women. Their mean age was 70.1 years. We compared the intervertebral angle, posterior disc height, and interspinous process distance on midsagittal magnetic resonance images obtained before and after the surgery. We also assessed clinical outcomes by using the Visual Analog Scale and the Zurich Claudication Questionnaire. The average operation time per level was 44.7 minutes. Postoperatively, there were significant changes in the angle (from 12.5 to 8.6 degrees, P < 0.0001), the posterior disc height (from 10.6 to 13.1 mm, P < 0.0001), and the interspinous process distance (from 9.7 to 14.1 mm, P < 0.0001). The clinical outcomes, which we assessed by using the Visual Analog Scale and the Zurich Claudication Questionnaire, were considered satisfactory. (Visual Analog Scale, from 6.88 to 3.00; Zurich Claudication Questionnaire, symptom severity domain from 2.94 to 1.92, physical function from 2.51 to 1.73.) Our ceramic spacer is useful in the treatment of elderly patients with lumbar canal stenosis. Treatment comprises an easy surgical procedure and produces no metal artifact on radiological evaluations, such as magnetic resonance imaging and computed tomographic scans.


Tissue engineering potential of human dermis-isolated adult stem cells from multiple anatomical locations.
Abundance and accessibility render skin-derived stem cells an attractive cell source for tissue engineering applications. Toward assessing their utility, the variability of constructs engineered from human dermis-isolated adult stem (hDIAS) cells was examined with respect to different anatomical locations (foreskin, breast, and abdominal skin), both in vitro and in a subcutaneous, athymic mouse model. All anatomical locations yielded hDIAS cells with multi-lineage differentiation potentials, though adipogenesis was not seen for foreskin-derived hDIAS cells. Using engineered cartilage as a model, tissue engineered constructs from hDIAS cells were compared. Construct morphology differed by location. The mechanical properties of human foreskin- and abdominal skin-derived constructs were similar at implantation, remaining comparable after 4 additional weeks of culture in vivo. Breast skin-derived constructs were not mechanically testable. For all groups, no signs of abnormality were observed in the host. Addition of aggregate redifferentiation culture prior to construct formation improved chondrogenic differentiation of foreskin-derived hDIAS cells, as evident by increases in glycosaminoglycan and collagen contents. More robust Alcian blue staining and homogeneous cell populations were also observed compared to controls. Human DIAS cells elicited no adverse host responses, reacted positively to chondrogenic regimens, and possessed multi-lineage differentiation potential with the caveat that efficacy may differ by anatomical origin of the skin. Taken together, these results suggest that hDIAS cells hold promise as a potential cell source for a number of tissue engineering applications.


Protocol for a prospective, observational cohort study of awareness in mechanically ventilated patients admitted from the emergency department: the ED-AWARENESS study.
Awareness with paralysis is a complication with potentially devastating psychological consequences for mechanically ventilated patients. While rigorous investigation into awareness has occurred for operating room patients, little attention has been paid outside of this domain. Mechanically ventilated patients in the emergency department (ED) have been historically managed in a way that predisposes them to awareness events: high incidence of neuromuscular blockade use, underdosing of analgesia and sedation, delayed administration of analgesia and sedation after intubation, and a lack of monitoring of sedation targets and depth. These practice patterns are discordant to recommendations for reducing the incidence of awareness, suggesting there is significant rationale to examine awareness in the ED population. This is a single centre, prospective cohort study examining the incidence of awareness in mechanically ventilated ED patients. A cohort of 383 mechanically ventilated ED patients will be included. The primary outcome is awareness with paralysis. Qualitative reports of all awareness events will be provided. Recognising the potential problem with conventional multivariable analysis arising from a small number of events (expected less than 10-phenomenon of separation), Firth penalised method, exact logistic regression model or penalised maximum likelihood estimation shrinkage (Ridge, LASSO) will be used to assess for predictors of awareness. Approval of the study by the Human Research Protection Office has been obtained. This work will be disseminated by publication of peer-reviewed manuscripts, presentation in abstract form at scientific meetings and data sharing with other investigators through academically established means.


Cytoplasmic RNA sequences complementary to cloned chick delta-crystallin cDNA show size heterogeneity.
Double-stranded complementary DNA (cDNA) sequences were prepared from day-old chick lens total polysomal RNA and inserted into the unique PstI restriction site of the plasmid pBR322. Colonies containing sequences complementary to abundant lens poly(A)-containing RNA sequences were identified by using lens 32P-labelled cDNA. Some of these clones have been characterized as containing delta-crystallin mRNA coding sequences by genomic DNA blot hybridization and RNA blot hybridizations. Hybridization of labelled DNA from such clones to RNA blots detected four size classes of delta-crystallin RNA sequences, although Southern blots indicated that there are probably only two delta-crystallin genes.


Rapidly infrared-assisted cooperatively self-assembled highly ordered multiscale porous materials.
In this paper, cooperative self-assembly (CSA) of colloidal spheres with different sizes was studied. It was found that a complicated jamming effect makes it difficult to achieve an optimal self-assembling condition for construction of a well-ordered stacking of colloidal spheres in a relatively short growth time by CSA. Through the use of a characteristic infrared (IR) technique to significantly accelerate local evaporation on the growing interface without changing the bulk growing environment, a concise three-parameter (temperature, pressure, and IR intensity) CSA method to effectively overcome the jamming effect has been developed. Mono- and multiscale inverse opals in a large range of lattice scales can be prepared within a growth time (15-30 min) that is remarkably shorter than the growth times of several hours for previous methods. Scanning electron microscopy images and transmittance spectra demonstrated the superior crystalline and optical qualities of the resulting materials. More importantly, the new method enables optimal conditions for CSA without limitations on sizes and materials of multiple colloids. This strategy not only makes a meaningful advance in the applicability and universality of colloidal crystals and ordered porous materials but also can be an inspiration to the self-assembly systems widely used in many other fields, such as nanotechnology and molecular bioengineering.


Effect of long-term treatment of 3T3-L1 adipocytes with chlorate on the synthesis, glycosylation, intracellular transport and secretion of lipoprotein lipase.
Lipoprotein lipase (LPL) is synthesized and glycosylated in the endoplasmic reticulum (ER), transported through the Golgi to the cell surface, and finally secreted. To examine the role of heparan sulphate proteoglycans (HSPG) in the synthesis, activity, intracellular transport and secretion of LPL, 3T3-L1 adipocytes were cultured for 7 days in the presence of 20 mM chlorate, an inhibitor of sulphation of HSPG. Treatment of cells with 20 mM chlorate for 7 days caused a 55% decrease in LPL activity in the intracellular compartment and a 79% decrease in the cell-surface compartment. The synthetic rate of LPL in chlorate-treated cells was identical with that in control cells as determined by biosynthetic labelling. The study with endoglycosidase H (endo H) showed that the treatment with chlorate increased the proportion of LPL subunits which were totally endo H-sensitive. The study with a heparin-Sepharose column showed that 3T3-L1 adipocytes contained three forms of LPL. The first form, accounting for 35% of the LPL, did not bind to the heparin-Sepharose column and had little or no activity; the second form, accounting for 32%, bound to the column and was eluted with 0.4-0.75 M NaCl but had no activity; the third form, accounting for 33%, bound to the column and was eluted with 0.8-1.2 M NaCl and had activity. In chlorate-treated cells, the first form accounted for 66% of the LPL, the second form 15% and the third form 19%. When cells were incubated for 1 h with brefeldin A, which translocates Golgi proteins to the ER [J. Lippincott-Schwartz, L.C. Yuan, J.S. Banifacino and R.D. Klausner (1989) Cell 56, 801-813; J. Lippincott-Schwartz, J. Glickman, J.E. Donaldson, J. Robbins, T.E. Kreis, K.B. Seamon, M.P. Sheetz and R.D. Klausner (1991) J. Cell Biol. 112, 567-577], the chlorate-induced decrease in cellular LPL activity was restored. These findings indicate that LPL synthesized in chlorate-treated cells can be processed to be fully active, but chlorate-treated cells are unable to transport LPL to the Golgi and accumulate inactive LPL with a lower affinity for heparin in the ER. The treatment with chlorate decreased the proportion of LPL subunits that were endo H-resistant, indicating that the processing of oligosaccharide chains of LPL in the trans-Golgi was impaired in chlorate-treated cells. The amount of 35S-labelled LPL secreted by chlorate-treated cells was identical with that secreted by control cells, whereas the level of LPL activity in the medium of chlorate-treated cells was 25% of that in the medium of control cells, indicating that most of the LPL secreted by chlorate-treated cells was inactive.


Presenilins are required for the formation of comma- and S-shaped bodies during nephrogenesis.
Mammalian presenilins consist of two highly homologous proteins, PSEN1 and PSEN2, which share redundant activities in Notch processing and signaling. To bypass the early lethality of the Psen1- and Psen2-double (PSEN) null embryos, we used a human PSEN1 transgene to rescue the somite patterning defects in PSEN-null animals and to allow a determination of the function of presenilins in late embryogenesis. We report here that expression of the human PSEN1 transgene supported the survival of PSEN-null embryos to the perinatal stage. However, presenilin deficiency in the kidney led to severe nephrogenesis defects and virtually no comma- or S-shaped bodies, or mature glomeruli were formed. We document that the mesenchyme was induced which could further progress to renal vesicles in the PSEN-null kidney, indicating that the presenilins are not essential for the inductive interactions and mesenchyme to epithelium transition. However, renal vesicles failed to pattern to form proximal tubules and glomerular epithelium. A presenilin-dependent, signaling-competent form of Notch1 was detected in mesenchymal derivatives but not in the ureteric buds of wild-type mice. Consistent with an obligatory role of presenilins in Notch processing and activation, the active form of Notch1 and its downstream target Hesr1 were absent in the PSEN-null kidney. Importantly, sustained Notch1 signaling was required for the maintenance of Notch ligand Jag1 expression. These results identify presenilins as one determinant of renal vesicle patterning in the developing mouse kidney, and we hypothesize that they act through the Notch signaling pathway.


Alternative tobacco product use and smoking cessation: a national study.
We investigated the frequency of alternative tobacco product use (loose leaf, moist snuff, snus, dissolvables, electronic cigarettes [e-cigarettes]) among smokers and the association with quit attempts and intentions. A nationally representative probability-based cross-sectional survey of 1836 current or recently former adult smokers was completed in November 2011. Multivariate logistic regressions evaluated associations between alternative tobacco product use and smoking cessation behaviors. Of the smokers, 38% had tried an alternative tobacco product, most frequently e-cigarettes. Alternative tobacco product use was associated with having made a quit attempt, and those intending to quit were significantly more likely to have tried and to currently use the products than were smokers with no intentions to quit. Use was not associated with successful quit attempts. Interest in future use of alternative tobacco products was low, except for e-cigarettes. Alternative tobacco products are attractive to smokers who want to quit smoking, but these data did not indicate that alternative tobacco products promote cessation. Unsubstantiated overt and implied claims that alternative tobacco products aid smoking cessation should be prohibited.


Hypochlorite and superoxide radicals can act synergistically to induce fragmentation of hyaluronan and chondroitin sulphates.
Activated phagocytes release the haem enzyme MPO (myeloperoxidase) and also generate superoxide radicals (O2*-), and hence H2O2, via an oxidative burst. Reaction of MPO with H2O2 in the presence of chloride ions generates HOCl (the physiological mixture of hypochlorous acid and its anion present at pH 7.4). Exposure of glycosaminoglycans to a MPO-H2O2-Cl- system or reagent HOCl generates long-lived chloramides [R-NCl-C(O)-R'] derived from the glycosamine N-acetyl functions. Decomposition of these species by transition metal ions gives polymer-derived amidyl (nitrogen-centred) radicals [R-N*-C(O)-R'], polymer-derived carbon-centred radicals and site-specific strand scission. In the present study, we have shown that exposure of glycosaminoglycan chloramides to O2*- also promotes chloramide decomposition and glycosaminoglycan fragmentation. These processes are inhibited by superoxide dismutase, metal ion chelators and the metal ion-binding protein BSA, consistent with chloramide decomposition and polymer fragmentation occurring via O2*--dependent one-electron reduction, possibly catalysed by trace metal ions. Polymer fragmentation induced by O2*- [generated by the superoxide thermal source 1, di-(4-carboxybenzyl)hyponitrite] was demonstrated to be entirely chloramide dependent as no fragmentation occurred with the native polymers or when the chloramides were quenched by prior treatment with methionine. EPR spin-trapping experiments using 5,5-dimethyl1-pyrroline-N-oxide and 2-methyl-2-nitrosopropane have provided evidence for both O2*- and polymer-derived carbon-centred radicals as intermediates. The results obtained are consistent with a mechanism involving one-electron reduction of the chloramides to yield polymer-derived amidyl radicals, which subsequently undergo intramolecular hydrogen atom abstraction reactions to give carbon-centred radicals. The latter undergo fragmentation reactions in a site-specific manner. This synergistic damage to glycosaminoglycans induced by HOCl and O2*- may be of significance at sites of inflammation where both oxidants are generated concurrently.


Genetic functions promoting homologous recombination in Escherichia coli: a study of inversions in phage lambda.
We have studied homologous recombination in a derivative of phage lambda containing two 1.4-kb repeats in inverted orientation. Inversion of the intervening 2.5-kb segment occurred efficiently by the Escherichia coli RecBC pathway but markedly less efficiently by the lambda Red pathway or the E. coli RecE or RecF pathways. Inversion by the RecBCD pathway was stimulated by Chi sites located to the right of the invertible segment; this stimulation decreased exponentially by a factor of about 2 for each 2.2 kb between the invertible segment and the Chi site. In addition to RecA protein and RecBCD enzyme, inversion by the RecBC pathway required single-stranded DNA binding protein, DNA gyrase, DNA polymerase I and DNA ligase. Inversion appeared to occur either intra- or intermolecularly. These results are discussed in the framework of a current molecular model for the RecBC pathway of homologous recombination.


Pharmaceutical agent cetylpyridinium chloride inhibits immune mast cell function by interfering with calcium mobilization.
Cetylpyridinium chloride (CPC) is an antimicrobial used in numerous personal care and janitorial products and food for human consumption at millimolar concentrations. Minimal information exists on the eukaryotic toxicology of CPC. We have investigated the effects of CPC on signal transduction of the immune cell type mast cells. Here, we show that CPC inhibits the mast cell function degranulation with antigen dose-dependence and at non-cytotoxic doses ∼1000-fold lower than concentrations in consumer products. Previously we showed that CPC disrupts phosphatidylinositol 4,5-bisphosphate, a signaling lipid critical for store-operated Ca 2+ entry (SOCE), which mediates degranulation. Our results indicate that CPC inhibits antigen-stimulated SOCE: CPC restricts Ca 2+ efflux from endoplasmic reticulum, reduces Ca 2+ uptake into mitochondria, and dampens Ca 2+ flow through plasma membrane channels. While inhibition of Ca 2+ channel function can be caused by alteration of plasma membrane potential (PMP) and cytosolic pH, CPC does not affect PMP or pH. Inhibition of SOCE is known to depress microtubule polymerization, and here we show that CPC indeed dose-dependently shuts down formation of microtubule tracks. In vitro data reveal that CPC inhibition of microtubules is not due to direct CPC interference with tubulin. In summary, CPC is a signaling toxicant that targets Ca 2+ mobilization.


Alpha v beta 3 integrin expression in human odontoblasts and co-localization with osteoadherin.
Integrins are heterodimeric transmembrane receptors which promote cell adhesion, thus contributing to the maintenance of tissue organization in both normal and pathological conditions. To characterize the way odontoblasts may interact with other cells and the extracellular matrix in human teeth, we studied expression of alpha v beta 3 integrin, a putative receptor for osteoadherin. We showed that alpha v beta 3 integrin expression was restricted to odontoblasts, blood vessels, and small rounded cells in sound and carious pulp. Odontoblast staining intensity increased from the apical to the cusp region. Osteoadherin staining was strong in the whole odontoblast layer (with a slight decrease in the cusp region) and in predentin. Odontoblasts differentiating in vitro were stained with the anti-alpha v beta 3 integrin antibody, first at the level of intercellular contacts, then throughout the cell membrane. These results suggest that the alpha v beta 3 integrin could play a role in interodontoblast adhesion and odontoblast binding to the surrounding predentin/dentin/pulp matrix, possibly through osteoadherin.


Childhood malnutrition: toward an understanding of infections, inflammation, and antimicrobials.
Undernutrition in childhood is estimated to cause 3.1 million child deaths annually through a potentiating effect on common infectious diseases, such as pneumonia and diarrhea. In turn, overt and subclinical infections, and inflammation, especially in the gut, alter nutrient intake, absorption, secretion, diversion, catabolism, and expenditure. A narrative overview of the current understanding of infections, inflammation, and antimicrobials in relation to childhood malnutrition. Searches for pivotal papers were conducted using PUBMED 1966-January 2013; hand searches of the references of retrieved literature; discussions with experts; and personal experience from the field. Although the epidemiological evidence for increased susceptibility to life-threatening infections associated with malnutrition is strong, we are only just beginning to understand some of the mechanisms involved. Nutritional status and growth are strongly influenced by environmental enteric dysfunction (EED), which is common among children in developing countries, and by alterations in the gut microbiome. As yet, there are no proven interventions against EED. Antibiotics have long been used as growth promoters in animals. Trials of antibiotics have shown striking efficacy on mortality and on growth in children with uncomplicated severe acute malnutrition (SAM) or HIV infection. Antibiotics act directly by preventing infections and may act indirectly by reducing subclinical infections and inflammation. We describe an ongoing multicenter, randomized, placebo-controlled trial of daily cotrimoxazole prophylaxis to prevent death in children recovering from complicated SAM. Secondary outcomes include growth, frequency and etiology of infections, immune activation and function, the gut microbiome, and antimicrobial resistance. The trial is expected to be reported in mid-2014. As well as improving nutritional intake, new case management strategies need to address infection, inflammation, and microbiota and assess health outcomes rather than only anthropometry.


Prenatal cocaine exposure: an examination of childhood externalizing and internalizing behavior problems at age 7 years.
This study examines the relationship between prenatal cocaine exposure and parent-reported child behavior problems at age 7 years. Data are from 407 African-American children (210 cocaine-exposed, 197 non-cocaine-exposed) enrolled prospectively at birth in a longitudinal study on the neurodevelopmental consequences of in utero exposure to cocaine. Prenatal cocaine exposure was assessed at delivery through maternal self-report and bioassays (maternal and infant urine and infant meconium). The Achenbach Child Behavior Checklist (CBCL), a measure of childhood externalizing and internalizing behavior problems, was completed by the child's current primary caregiver during an assessment visit scheduled when the child was seven years old. Structural equation and GLM/GEE models disclosed no association linking prenatal cocaine exposure status or level of cocaine exposure to child behavior (CBCL Externalizing and Internalizing scores or the eight CBCL subscale scores). This evidence, based on standardized ratings by the current primary caregiver, fails to support hypothesized cocaine-associated behavioral problems in school-aged children with in utero cocaine exposure. A next step in this line of research is to secure standardized ratings from other informants (e.g., teachers, youth self-report).


Finding patterns in emotional information: Enhanced sensitivity to statistical regularities within negative information.
In everyday life, people are exposed to continuous flows of emotional information. The ability to organize and segment this continuous input seems critical to understand what is happening around us. This study investigated whether people are sensitive to subtle statistical regularities embedded in flows of emotional information. Experiment 1 showed that people were able to identify regularities in streams of negative visual scenes. Experiment 2 demonstrated that the ability to extract statistical regularities was enhanced for negative compared to neutral visual scenes. Finally, Experiment 3 found that learning of statistical regularities was similar for negative and positive visual scenes. Across these experiments, we found that explicit awareness of regularities did not modulate statistical learning ability. The results of this study help to understand the processes that enable people to make inferences about the complex input of the emotional world. (PsycInfo Database Record (c) 2020 APA, all rights reserved).


Skin tumours induced by narrowband UVB have higher frequency of p53 mutations than tumours induced by broadband UVB independent of Ogg1 genotype.
Different wavelengths of ultraviolet (UV) light have different promoting effects on skin carcinogenesis. Narrowband UVB (NB-UVB) has a single-peak wavelength of 311 nm and is widely used for treating skin diseases. Our previous work showed that, in comparison with conventional broadband UVB (BB-UVB), long-term exposure to NB-UVB induces higher frequency of skin cancer in mice, and it suggested that this is mediated through the formation of cyclobutane pyrimidine dimers (CPDs). To explore whether the frequency of p53 mutations in skin tumours correlates with CPD-induced mutations, we compared the frequency and types of p53 mutations between NB-UVB-induced and BB-UVB-induced malignant skin tumours produced in wild-type and Ogg1 knockout mice, which are deficient in repair of oxidative 8-oxoguanine (8-oxoG), a DNA damage mediated by reactive oxygen species (ROS). The frequency of p53 mutation was significantly higher in NB-UVB-induced than in BB-UVB-induced tumours in both wild-type and Ogg1 knockout mice. Most of the p53 mutations found were G:C → A:T transitions at dipyrimidine sites in both the NB-UVB- and BB-UVB-exposed groups. However, G:C → T:A mutations caused by 8-oxoG did not increase in Ogg1 knockout mice exposed to either NB-UVB or BB-UVB. Our results strongly suggest that NB-UVB induces highly malignant tumours caused by p53 dipyrimidine mutations through the formation of CPDs.


Bone Marrow-Derived GCA+ Immune Cells Drive Alzheimer's Disease Progression.
Alzheimer's disease (AD) is an age-related degenerative disease of the central nervous system (CNS), whereas the role of bone marrow immune cells in the pathogenesis of AD remains unclear. Here, the study reveals that compared to matched healthy individuals, AD patients have higher circulating grancalcin (GCA) levels, which negatively correlate with cognitive function. Bone marrow-derived GCA+ immune cells, which secret abundant GCA and increase during aging, preferentially invaded the hippocampus and cortex of AD mouse model in a C-C Motif Chemokine Receptor 10 (CCR10)-dependent manner. Transplanting GCA+ immune cells or direct stereotaxic injection of recombinant GCA protein intensified amyloid plaque load and aggravated cognitive and memory impairments. In contrast, genetic ablation of GCA in the hematopoietic compartment improves cognitive and memory function. Mechanistically, GCA competitively binds to the low-density lipoprotein receptor-related protein 1 (LRP1) in microglia, thus inhibiting phagocytosis and clearance of Aβ and potentiating neuropathological changes. Importantly, GCA-neutralizing antibody treatment rejuvenated cognitive and memory function and constrained AD progression. Together, the study demonstrates a pathological role of GCA+ immune cells instigating cognitive and memory decline, suggesting that GCA+ immune cells can be a potential target for innovative therapeutic strategies in AD.


A decision analysis of the effectiveness of the pediatric telephone triage program in Japan.
Pediatric telephone triage programs have been initiated to reduce overcrowding of pediatric medical facilities, yet it is unclear what impact these programs have on reducing after hours room visits. This study used a decision analysis model and data from reference literature to investigate the effectiveness of this program and determine the factors that influence the results. The decision analysis model focused on a hypothetical group of children who were not undergoing ongoing treatment but developed symptoms or sustained injuries thought to require a nighttime visit to a medical facility. The model differentiated between urgent, semi-urgent, and non-urgent cases. Model parameters were estimated from the literature whenever possible. We estimated the difference in the number of children who received emergency medical attention, the number of semi-urgent cases in which attention was delayed, and the cases that did not receive medical care, between the telephone triage group and the control group. Telephone triage reduced the number of after hours visits by approximately 4%. There was no change in the number of semi-urgent cases for which medical care was delayed, and the number of semi-urgent cases that did not receive medical attention increased by three. The sensitivity analysis showed that increasing telephone triage use from 10% to 20% would reduce after hours visits by approximately 8%. We conclude that the benefits of this program to the region as a whole would outweigh the harmful effects. However, the effectiveness of telephone triage programs will be limited unless its usage is expanded.


Residual Spinal Cord Compression Following Hemilaminectomy and Mini-Hemilaminectomy in Dogs: A Prospective Randomized Study.
The aim of this study was to compare the reduction of spinal cord compression after surgical treatment of dogs with acute thoracolumbar intervertebral disc (IVD) extrusion achieved using hemilaminectomy versus mini-hemilaminectomy techniques. This was a prospective randomized study with client-owned dogs presented with acute IVD extrusion that were allocated to surgical treatment using hemilaminectomy (n = 15) or mini-hemilaminectomy (n = 15) techniques. Plain and intravenous-contrast computed tomography was performed pre- and postoperatively. The preoperative minimal cross-sectional dimension of the spinal cord (MDSCpre) and the postoperative minimal cross-sectional dimension of the spinal cord (MDSCpost) were measured at the level of greatest compression. The minimal diameter of the uncompressed spinal cord was measured in a similar way both pre- (MDUSCpre) and postoperatively (MDUSCpost). Dogs in the mini-hemilaminectomy group had significantly greater reduction of compression (RC) (p < 0.01) after surgery compared to dogs in the hemilaminectomy group. The mean RC in the hemilaminectomy group was 34.6% and in the mini-hemilaminectomy group 62.6%. Our results showed a significantly greater reduction of spinal cord compression for mini-hemilaminectomy compared to hemilaminectomy. Additionally, mini-hemilaminectomy could be a preferred method due to its minimal invasiveness and easier access to lateral fenestration.


PCP4 regulates Purkinje cell excitability and cardiac rhythmicity.
Cardiac Purkinje cells are important triggers of ventricular arrhythmias associated with heritable and acquired syndromes; however, the mechanisms responsible for this proarrhythmic behavior are incompletely understood. Here, through transcriptional profiling of genetically labeled cardiomyocytes, we identified expression of Purkinje cell protein-4 (Pcp4), a putative regulator of calmodulin and Ca2+/calmodulin-dependent kinase II (CaMKII) signaling, exclusively within the His-Purkinje network. Using Pcp4-null mice and acquired cardiomyopathy models, we determined that reduced expression of PCP4 is associated with CaMKII activation, abnormal electrophysiology, dysregulated intracellular calcium handling, and proarrhythmic behavior in isolated Purkinje cells. Pcp4-null mice also displayed profound autonomic dysregulation and arrhythmic behavior in vivo. Together, these results demonstrate that PCP4 regulates cardiac excitability through both Purkinje cell-autonomous and central mechanisms and identify this modulator of CaMKII signaling as a potential arrhythmia-susceptibility candidate.


Hereditary laminitis in Jersey calves in Zimbabwe.
The clinical signs and radiological findings of a rare laminitis-like condition in Jersey calves (n = 6) are described. Regular hoof-trimming proved very beneficial. Pedigree studies of the affected calves strongly suggest a recessive autosomal inheritance.


Plastid protein synthesis is required for plant development in tobacco.
Chloroplasts fulfill important functions in cellular metabolism. The majority of plastid genome-encoded genes is involved in either photosynthesis or chloroplast gene expression. Whether or not plastid genes also can determine extraplastidic functions has remained controversial. We demonstrate here an essential role of plastid protein synthesis in tobacco leaf development. By using chloroplast transformation, we have developed an experimental system that produces recombination-based knockouts of chloroplast translation in a cell-line-specific manner. The resulting plants are chimeric and, in the presence of translational inhibitors, exhibit severe developmental abnormalities. In the absence of active plastid protein synthesis, leaf blade development is abolished because of an apparent arrest of cell division. This effect appears to be cell-autonomous in that adjacent sectors of cells with translating plastids are phenotypically normal but cannot complement for the absence of plastid translation in mutant sectors. Developmental abnormalities also are seen in flower morphology, indicating that the defects are not caused by inhibited expression of plastid photosynthesis genes. Taken together, our data point to an unexpected essential role of plastid genes and gene expression in plant development and cell division.


Isometric exercise and cognitive function: an investigation of acute dose-response effects during submaximal fatiguing contractions.
The purpose of this study was to explore the dose-response relationship between exercise and cognitive performance using an acute bout of isometric exercise. University students (N = 55) were randomly assigned to control, 30%, 50% and 70% of maximum voluntary handgrip contraction groups. Participants performed a modified Stroop task before and after completion of an isometric handgrip endurance trial at their assigned exercise intensity. Ratings of perceived exertion (RPE) and forearm muscle activation (EMG) showed linear trends of progressively greater RPE and muscle activation at greater exercise intensity levels. Regression analysis showed significant (P < .05) linear degradations in frequency of errors on the Stroop task with increasing exercise intensity. We conclude that performing isometric exercise until exhaustion is associated with reduced cognitive performance and that higher intensity isometric exercise leads to greater performance impairments in a linear dose-response manner.


Newer emerging pathogens of ocular non-sporulating molds (NSM) identified by polymerase chain reaction (PCR)-based DNA sequencing technique targeting internal transcribed spacer (ITS) region.
To apply Polymerase Chain Reaction (PCR)-based DNA sequencing targeting Internal Transcribed Spacer (ITS) region for identification of non-sporulating molds (NSM) to species level which formed 12% of ocular isolates of fungi in a tertiary eye hospital in South India. Fifty ocular filamentous fungal NSM isolates recovered from 45 patients were included in the study. PCR-based DNA sequencing technique targeting ITS region was applied to identify NSM. PCR-based DNA sequencing revealed 23 established pathogens involving 8 genera (Aspergillus, Fusarium, Bipolaris, Pythium, Cochilobolus, Exserohilum, Pseudoallescheria, and Scedosporiumspecies) and 27 emerging pathogens involving 7 genera (Botryosphaeria Lasiodiplodia species, Thielavia tortuosa, Glomerulla singulata, Macrophomina phaseolina, Rhizoctonia bataticola, and Podosporaspecies) reported for the first time in literature related to ocular infections. Fifteen (30%) patients with fungal keratitis caused by NSM failed to respond to standard antifungal therapy. PCR-based DNA sequencing technique is a rapid, reliable, and valuable tool to identify 54% of NSM as newer potential pathogens of fungi causing ocular mycoses.


Effect of local application of epidermal growth factor on innate immunity and cell composition of destruction focus in experimental thermal injury.
The effect of recombinant human epidermal growth factor (rhEGF) on innate immunity and cellular composition of the destruction focus in the third-degree (IIIA) burn (skin contact with an object heated to 100°C; 4% body surface) was studied in experiments on outbred albino rats. On days 7-28 after burn, blood count of phagocytes and their absorbing capacity and oxygen-dependent metabolism increased, which correlated with the increase in serum IL-1β level and neutrophil count in the destruction focus. Local application of rhEGF led to earlier (on day 14) normalization of the count and functional activity of blood phagocytes and decrease in serum IL-1β level and accelerated neutrophil and lymphocyte replacement with fibroblasts in the focus of injury.


Nationwide risk factors for hospital readmission for subsequent injury after motor vehicle crashes.
Some drivers involved in motor vehicle crashes across the United States may be identified as at risk of subsequent injury by a similar mechanism. The purpose of this study was to perform a national review of the risk factors for hospitalization for a new injury due to a subsequent motor vehicle crash. It was hypothesized that presenting to a different hospital after subsequent injury would result in worse patient outcomes when compared to presentation at the same hospital. The Nationwide Readmissions Database for 2010-2014 was queried for all inpatient hospitalizations with injury related to motor vehicle traffic. The primary patient outcome of interest was subsequent motor vehicle crash-related injury within 1 year. The secondary patient outcomes were different hospital subsequent injury presentation, higher Injury Severity Score (ISS), longer length of stay (LOS), and in-hospital death after subsequent injury. The analysis of secondary patient outcomes was performed only on patients who were reinjured. Univariable analysis was performed for each outcome using all variables during the index admission. Multivariable logistic regression was performed using all significant (P < .05) variables on univariate analysis. Results were weighted for national estimates. During the study period, 1,008,991 patients were admitted for motor vehicle-related injury; 12,474 patients (1.2%) suffered a subsequent injury within 1 year. From the reinjured patients, 32.9% presented to a different hospital, 48.9% had a higher ISS, and 22.1% had a longer LOS. The in-hospital mortality rate after subsequent injury was 1.1%. Presentation to a different hospital for subsequent injury was associated with a longer LOS (odds ratio [OR] = 1.32; 95% confidence interval [CI], 1.20-1.45; P < .01) and a higher ISS (OR = 1.38; 95% CI, 1.27-1.49; P < .01). Motorcyclists were more likely to suffer subsequent injury (OR = 1.39; 95% CI, 1.32-1.46; P < .01) and motorcycle passengers were more likely to present to a different hospital with a subsequent injury (OR = 2.49; 95% CI, 1.73-3.59; P < .01). Alcohol abuse was associated with subsequent injury (OR = 1.12; 95% CI, 1.07-1.18; P < .01). Nearly a third of patients suffering subsequent motor vehicle crash-related injury after an initial motor vehicle crash in the United States present to a different hospital. These patients are more likely to suffer more severe injuries and longer hospitalizations due to their subsequent injury. Future efforts to prevent these injuries must consider the impact of this fragmentation of care and the implications for quality and cost improvements.


A cross-sectional study examining Campylobacter and other zoonotic enteric pathogens in dogs that frequent dog parks in three cities in south-western Ontario and risk factors for shedding of Campylobacter spp.
An estimated 6 million pet dogs live in Canadian households with the potential to transmit zoonotic pathogens to humans. Dogs have been identified as carriers of Salmonella, Giardia and Campylobacter spp., particularly Campylobacter upsaliensis, but little is known about the prevalence and risk factors for these pathogens in pet dogs that visit dog parks. This study examined the prevalence of these organisms in the faeces of dogs visiting dog parks in three cities in south-western Ontario, as well as risk factors for shedding Campylobacter spp. and C. upsaliensis. From May to August 2009, canine faecal samples were collected at ten dog parks in the cities of Guelph and Kitchener-Waterloo, Ontario, Canada. Owners were asked to complete a questionnaire related to pet characteristics and management factors including age, diet and activities in which the dog participates. Faecal samples were collected from 251 dogs, and 189 questionnaires were completed. Salmonella, Giardia and Campylobacter spp. were present in 1.2%, 6.4% and 43.0% of faecal samples, respectively. Of the Campylobacter spp. detected, 86.1% were C. upsaliensis, 13% were C. jejuni and 0.9% were C. coli. Statistically significant sparing factors associated with the shedding of Campylobacter spp. included the feeding of a commercial dry diet and the dog's exposure to compost. Age of dog had a quadratic effect, with young dogs and senior dogs having an increased probability of shedding Campylobacter spp. compared with adult dogs. The only statistically significant risk factor for shedding C. upsaliensis was outdoor water access including lakes and ditches, while dogs >1 year old were at a lower risk than young dogs. Understanding the pet-related risk factors for Campylobacter spp. and C. upsaliensis shedding in dogs may help in the development of awareness and management strategies to potentially reduce the risk of transmitting this pathogen from dogs to humans.


WRKY Gene Family Drives Dormancy Release in Onion Bulbs.
Onion (Allium cepa L.) is an important bulb crop grown worldwide. Dormancy in bulbous plants is an important physiological state mainly regulated by a complex gene network that determines a stop of vegetative growth during unfavorable seasons. Limited knowledge on the molecular mechanisms that regulate dormancy in onion were available until now. Here, a comparison between uninfected and onion yellow dwarf virus (OYDV)-infected onion bulbs highlighted an altered dormancy in the virus-infected plants, causing several symptoms, such as leaf striping, growth reduction, early bulb sprouting and rooting, as well as a lower abscisic acid (ABA) level at the start of dormancy. Furthermore, by comparing three dormancy stages, almost five thousand four hundred (5390) differentially expressed genes (DEGs) were found in uninfected bulbs, while the number of DEGs was significantly reduced (1322) in OYDV-infected bulbs. Genes involved in cell wall modification, proteolysis, and hormone signaling, such as ABA, gibberellins (GAs), indole-3-acetic acid (IAA), and brassinosteroids (BRs), that have already been reported as key dormancy-related pathways, were the most enriched ones in the healthy plants. Interestingly, several transcription factors (TFs) were up-regulated in the uninfected bulbs, among them three genes belonging to the WRKY family, for the first time characterized in onion, were identified during dormancy release. The involvement of specific WRKY genes in breaking dormancy in onion was confirmed by GO enrichment and network analysis, highlighting a correlation between AcWRKY32 and genes driving plant development, cell wall modification, and division via gibberellin and auxin homeostasis, two key processes in dormancy release. Overall, we present, for the first time, a detailed molecular analysis of the dormancy process, a description of the WRKY-TF family in onion, providing a better understanding of the role played by AcWRKY32 in the bulb dormancy release. The TF co-expressed genes may represent targets for controlling the early sprouting in onion, laying the foundations for novel breeding programs to improve shelf life and reduce postharvest.


The influence of abciximab use on clinical outcome after aortocoronary vein graft interventions.
The purpose of this study was to evaluate the effect of abciximab use on clinical outcome in aortocoronary vein graft interventions. Although large randomized trials have demonstrated a significant benefit of abciximab use in the setting of percutaneous coronary interventions, there is relatively little data with respect to the use of this agent in percutaneous vein graft interventions. Three hundred and forty-three patients were identified; 210 undergoing vein graft intervention without abciximab and 133 patients with abciximab. There were differences in baseline clinical and angiographic characteristics between the two groups; advanced age, unstable angina, older vein grafts and thrombus containing lesions were relatively common in both groups. Angiographic and procedural success rates were similar with or without the use of abciximab (89% vs. 92%, p = 0.15, and 85% vs. 91%, p = 0.12, respectively). The in-hospital composite end point of death/Q-wave myocardial infarction (QWMI)/repeat revascularization was similar between the two groups. Utilizing statistical modeling to adjust for baseline differences between the groups, abciximab use did not influence the cumulative long-term composite end point of death/MI/repeat revascularization. This study demonstrates that in this relatively high-risk population undergoing aortocoronary vein graft interventions, the administration of abciximab periprocedurally does not appear to reduce major adverse clinical events.


Occurrence and Chemistry of Tropane Alkaloids in Foods, with a Focus on Sample Analysis Methods: A Review on Recent Trends and Technological Advances.
Tropane alkaloids (TAs) are natural toxins produced by different plants, mainly from the Solanaceae family. The interest in TAs analysis is due to the serious cases of poisoning that are produced due to the presence of TA-producing plants in a variety of foods. For this reason, in recent years, different analytical methods have been reported for their control. However, the complexity of the matrices makes the sample preparation a critical step for this task. Therefore, this review has focused on (a) collecting the available data in relation to the occurrence of TAs in foods for human consumption and (b) providing the state of the art in food sample preparation (from 2015 to today). Regarding the different food categories, cereals and related products and teas and herbal teas have been the most analyzed. Solid-liquid extraction is still the technique most widely used for sample preparation, although other extraction and purification techniques such as solid-phase extraction or QuEChERS procedure, based on the use of sorbents for extract or clean-up step, are being applied since they allow cleaner extracts. On the other hand, new materials (molecularly imprinted polymers, mesostructured silica-based materials, metal-organic frameworks) are emerging as sorbents to develop effective extraction and purification methods that allow lower limits and matrix effects, being a future trend for the analysis of TAs.


Characteristics of the polyriboinosinic acid:polyribocytidylic acid assay for noncytopathogenic bovine viral diarrhea virus.
The use of polyriboinosinic acid:polyribocytidylic acid (poly I:C) for noncytopathogenic bovine viral diarrhea virus (NC-BVDV) assay (PINBA) was studied. Several viruses were tested for their suitability as test challenge viruses. In addition to vesicular stomatitis virus, which previously was shown to be a suitable challenge virus, bovine enteroviruses also were found to be suitable, whereas infectious bovine rhinotracheitis virus and parainfluenza type 3 virus were only marginally suitable. Bovine embryonic skin (BES) cultures developed resistance to PINBA within a few in vitro passages, whereas bovine embryonic lung (BEL) cultures did not. At certain passages, BES cultures were 500,000 times more resistant than BEL cultures. To determine whether the difference in viral titers on BEL and BES cultures was due to NC-BVDV replication, PINBA and fluorescent antibody assays were compared on the cultures. Resistance of BES cultures to PINBA was not due to an inability of the virus to replicate in the cultures, but was due to an inability of PINBA to detect the virus. Viral titers were comparable by fluorescent antibody assay titers on BES and BEL cultures, but were considerably higher than viral titers on BES cultures with PINBA. Variations in viral titers, using PINBA on BEL cultures, were observed and were considered to be due to cultural conditions, such as the presence of low levels of BVDV antibodies in bovine fetal serum used in the medium. Treatment of BEL cultures with poly I:C or interferon showed that NC-BVDV was sensitive to interferon as determined by virus-yield reduction.


Association between light at night, melatonin secretion, sleep deprivation, and the internal clock: Health impacts and mechanisms of circadian disruption.
Exposure to Artificial Light At Night (ALAN) results in a disruption of the circadian system, which is deleterious to health. In industrialized countries, 75% of the total workforce is estimated to have been involved in shift work and night work. Epidemiologic studies, mainly of nurses, have revealed an association between sustained night work and a 50-100% higher incidence of breast cancer. The potential and multifactorial mechanisms of the effects include the suppression of melatonin secretion by ALAN, sleep deprivation, and circadian disruption. Shift and/or night work generally decreases the time spent sleeping, and it disrupts the circadian time structure. In the long run, this desynchronization is detrimental to health, as underscored by a large number of epidemiological studies that have uncovered elevated rates of several diseases, including cancer, diabetes, cardiovascular risks, obesity, mood disorders and age-related macular degeneration. It amounts to a public health issue in the light of the very substantial number of individuals involved. The IARC has classified shift work in group 2A of "probable carcinogens to humans" since "they involve a circadian disorganization". Countermeasures to the effects of ALAN, such as melatonin, bright light, or psychotropic drugs, have been proposed as a means to combat circadian clock disruption and improve adaptation to shift and night work. We review the evidence for the ALAN impacts on health. Furthermore, we highlight the importance of an in-depth mechanistic understanding to combat the detrimental properties of exposure to ALAN and develop strategies of prevention.


The pathophysiology of renin release in renovascular hypertension.
Renovascular hypertension (RVH) results from occlusion of blood flow to either kidney, which stimulates renin release. Increased renin leads to a series of actions that rapidly leads to increased systemic blood pressure. Experimental renovascular hypertension is developed in animals by placement of a clip that occludes more than 50% of renal blood flow to that kidney. The major stimulus for renin release in renovascular hypertension is the severe drop in hydrostatic pressure in the afferent arteriole, the location of the juxtaglomerular renin-secreting granular cells. The pressure drop changes the degree of stretch of these cells which leads to baroreceptor-mediated renin release. The level of renin released can be modified by sympathetic nerves and to a lesser degree by the macula densa. Several hormone or vasoactive agents may augment renin released during RVH, but nearly all are secondary to changes in the pressure receptor mechanism.