[Comparative study of clinicopathological features, and risk factors of advanced fibrosis between genders with non-alcoholic fatty liver disease].
Objective: To comparatively study the similarities and differences between the clinical, pathological, and risk factors of advanced fibrosis in men and women with non-alcoholic fatty liver disease (NAFLD). Methods: 267 patients with NAFLD diagnosed by liver biopsy were retrospectively included, and were divided into male and female groups. The difference of clinical and pathological indexes were compared between the two groups. The measurement data were in accordance with normal distribution. The comparison between the two groups was performed by independent sample t-test. The non-parametric test was used for non-normal distribution. The classification data were expressed as a percentage, and the chi-square test was used for comparison between groups. Logistic regression analysis was used to analyze the risk factors. Results: The age of onset of NAFLD was significantly lower in male than female patients (P < 0.01). There was no statistically significant difference between the male and female groups in terms of body mass index and the prevalence of type 2 diabetes (P > 0.05). Biochemical index: The levels of alanine aminotransferase, albumin, total bilirubin and uric acid were significantly higher in male than female patients (P < 0.01). Liver pathology: The proportion of ballooning degeneration was significantly lower in male than female patients (P < 0.01). There was not statistically significant difference between the two groups in the proportion of steatohepatitis score, non-alcoholic steatohepatitis (52.0% vs. 61.5%, P = 0.283) and advanced liver fibrosis (14.3% vs. 17.8%, P = 0.162). Thrombocytopenia was a common independent risk factor for advanced stage liver fibrosis (OR = 0.984, 0.978~0.989, P < 0.01). Type 2 diabetes was only an independent risk factor for advanced stage liver fibrosis in men (OR = 6.557, 1.667~25.782), P < 0.01). Elevated AST was only an independent risk factor for advanced stage liver fibrosis in women (OR = 1.016, 1.003~1.028, P = 0.012). Conclusion: In NAFLD patients, there are some clinical and pathological differences between genders. Platelets are a common predictor of advanced liver fibrosis in men and women. Type 2 diabetes in men and elevated aspartate aminotransferase in women can be regarded as independent risk factors for advanced liver fibrosis.


Ideological profiles of US adults and their support for youth mentoring.
Little is known about the influence of political ideology and religiosity on adults' support for youth mentoring as a strategy to address social problems. This study used latent class analysis in a large, national sample of US adults to identify underlying ideological profiles associated with support for mentoring programs. Three latent classes emerged. The attitudes of two classes, Classic Conservatives and Progressives, were consistent with traditional political conservatism and liberalism; the latter endorsed higher support for the theory of mentoring and government spending on mentoring programs. Members of the third class, Religious Outsiders, were highly religious, self-identified as very conservative, and were highly supportive of the theory of mentoring and the use of government funds on mentoring programs. Ad hoc analyses revealed that Religious Outsiders were the most likely to actually participate in mentoring activities. These findings suggest that support for mentoring, while not universal, crosses traditional political lines.


Brain tumor segmentation based on local independent projection-based classification.
Brain tumor segmentation is an important procedure for early tumor diagnosis and radiotherapy planning. Although numerous brain tumor segmentation methods have been presented, enhancing tumor segmentation methods is still challenging because brain tumor MRI images exhibit complex characteristics, such as high diversity in tumor appearance and ambiguous tumor boundaries. To address this problem, we propose a novel automatic tumor segmentation method for MRI images. This method treats tumor segmentation as a classification problem. Additionally, the local independent projection-based classification (LIPC) method is used to classify each voxel into different classes. A novel classification framework is derived by introducing the local independent projection into the classical classification model. Locality is important in the calculation of local independent projections for LIPC. Locality is also considered in determining whether local anchor embedding is more applicable in solving linear projection weights compared with other coding methods. Moreover, LIPC considers the data distribution of different classes by learning a softmax regression model, which can further improve classification performance. In this study, 80 brain tumor MRI images with ground truth data are used as training data and 40 images without ground truth data are used as testing data. The segmentation results of testing data are evaluated by an online evaluation tool. The average dice similarities of the proposed method for segmenting complete tumor, tumor core, and contrast-enhancing tumor on real patient data are 0.84, 0.685, and 0.585, respectively. These results are comparable to other state-of-the-art methods.


Hurricane effects, mitigation, and preparedness in the Caribbean: Perspectives on high importance-low prevalence practices from agricultural advisors.
Frequent hurricanes affect agricultural productivity, food security, economic security, and human wellbeing in the Caribbean islands. We assessed recent hurricane effects on the agricultural sector, and the challenges faced by farmers, foresters, and advisors related to hurricane preparedness and recovery in Puerto Rico and US Virgin Islands (USVI). We used qualitative and quantitative survey methods to compile perspectives from agricultural advisors related to hurricane effects on farmlands, preparedness and recovery measures taken by land managers, and the needs regarding preparing for and responding to future hurricanes. Survey responses from over 200 advisors at eight institutions provided insight into the most devastating hurricane effects across farmlands, including issues related to power outages, communication, road access, and fallen trees. Our results highlight strategies considered critical for hurricane preparedness and recovery but not prevalent in application among land managers. Advisors' perceptions suggested that farmers and ranchers apply essential recovery practices, but critical short-term preparedness practices are limited, and long-term preparedness practices are uncommon. Advisors also indicated the need for more training and educational resources to improve hurricane recovery response. We conclude that better planning to minimize the vulnerability to future hurricanes can be achieved through an increased understanding of how preparedness and recovery practices help mitigate hurricane effects, improved interagency coordination for hurricane response and preparedness, and integrated educational campaigns with advisors and land managers.


Clinical dementia rating: a reliable and valid diagnostic and staging measure for dementia of the Alzheimer type.
Global staging measures for dementia of the Alzheimer type (DAT) assess the influence of cognitive loss on the ability to conduct everyday activities and represent the "ultimate test" of efficacy for antidementia drug trials. They provide information about clinically meaningful function and behavior and are less affected by the "floor" and "ceiling" effects commonly associated with psychometric tests. The Washington University Clinical Dementia Rating (CDR) is a global scale developed to clinically denote the presence of DAT and stage its severity. The clinical protocol incorporates semistructured interviews with the patient and informant to obtain information necessary to rate the subject's cognitive performance in six domains: memory, orientation, judgment and problem solving, community affairs, home and hobbies, and personal care. The CDR has been standardized for multicenter use, including the Consortium to Establish a Registry for Alzheimer's Disease (CERAD) and the Alzheimer's Disease Cooperative Study, and interrater reliability has been established. Criterion validity for both the global CDR and scores on individual domains has been demonstrated, and the CDR also has been validated neuropathologically, particularly for the presence or absence of dementia. Standardized training protocols are available. Although not well suited as a brief screening tool for population surveys of dementia because the protocol depends on sufficient time to conduct interviews, the CDR has become widely accepted in the clinical setting as a reliable and valid global assessment measure for DAT.


Derivatives of dicyclopentadiene in ground water.
Dicyclopentadiene, a waste product of manufacturing at the Rocky Mountain Arsenal near Denver Colorado, has been detected in ground water at this facility. Ground water extracts were analysed by gas chromatography-mass spectrometry (GC/MS) to determine if derivatives of dicyclopentadiene were present in addition to dicyclopentadiene and other chemical wastes. The derivatives thus identified were characterized by GC high resolution MS, deuterium exchange for active hydrogen in the chemical ionization source of the mass spectrometer, and GC/MS following on-column base catalyzed deuteration of enolizable hydrogen. Two ketone derivatives of dicyclopentadiene were identified by comparison with synthetic standards. Ground water derivatives of dicyclopentadiene were compared with mammalian enzymatic metabolites produced in vitro by incubation with immobilized rabbit liver cytochromes P-450. The metabolites were analysed by GC/MS and by GC/MS following derivatization and deuterium labelling. Although not found in ground water samples, the two metabolites were identified as monoepoxides of dicyclopentadiene by comparison with compounds synthesized in the laboratory.


Premature termination codons destabilize ferredoxin-1 mRNA when ferredoxin-1 is translated.
Ferredoxin-1 (Fed-1) mRNA is poorly translated in dark-treated tobacco (Nicotiana tabacum) leaves, resulting in destabilization of Fed-1 mRNA and a differential light/dark accumulation of the mRNA. Insertion of nonsense codons within the Fed-1 coding sequence disrupts the light regulation of Fed-1 mRNA abundance. Here we show that the nonsense codon effect results primarily from lowering the Fed-1 mRNA stability in light-treated leaf tissue and in rapidly growing tobacco cell cultures, but not in dark-treated leaf tissue. These results suggest that nonsense codons trigger a decay pathway distinct from that seen for Fed-1 mRNA in the dark. We propose that nonsense-mediated decay of nonsense-containing Fed-1 mRNA occurs in light-treated leaves and in non-photosynthetic tobacco culture cells where Fed-1 mRNA is being actively translated.


Novel curcumin analog (cis-trans curcumin) as ligand to adenosine receptors A2A and A2B: potential for therapeutics.
All four of the adenosine receptor (AR) subtypes mediate pain and have been targeted by pharmacologists to generate new therapeutics for chronic pain. The vanilloid phytochemicals, which include curcumin, capsaicin, and gingerol, have been shown to alleviate pain. However, there is little to no literature on the interaction of vanilloid phytochemicals with ARs. In this study, photochemical methods were used to generate a novel isomer of curcumin (cis-trans curcumin or CTCUR), and the interactions of both curcumin and CTCUR with the two Gs-linked AR subtypes were studied. Competitive binding assays, docking analysis, and confocal fluorescence microscopy were performed to measure binding affinity; cell survival assays were used to measure toxicity; and cAMP assays were performed to measure receptor activation. Competitive binding results indicated that CTCUR binds to both AR A2A and AR A2B with Ki values of 5 μM and 7 μM, respectively, which is consistent with our docking results. Fluorescence microscopy data also shows binding for A2B and A2A. Cell survival results show that CTCUR and CUR are nontoxic at the tested concentrations in these cell lines. Overall, our results suggest that vanilloid phytochemicals may be slightly modified to increase interaction with Gs-ARs, and thereby can be further explored to provide a novel class of non-opioid antinociceptives.


Cancer patients seeking a second surgical opinion: results of a study on motives, needs, and expectations.
To explore the sociodemographic and clinical characteristics of cancer patients seeking a second-opinion consultation and to analyze their second opinion-related motives, needs, and expectations. In 212 consecutive patients seeking a second opinion at the Surgical Oncology Outpatient Clinic, satisfaction with the first specialist, motivation for the second opinion, need for information, preference for decision participation, and hope for and expectation of a different second opinion were assessed with a questionnaire. The mean age was 53 years. Most patients were women (82%), of whom 76% were diagnosed with breast cancer. Half of the patients (51%) had a low educational level. The majority of patients (62%) only had internal motives for second-opinion seeking associated with the need for reassurance and more certainty, whereas a substantial minority of patients (38%) also had external motives related to negative experiences or unfulfilled needs. The externally motivated patients had a higher anxiety disposition, were less satisfied with their first specialist, preferred a more active role in medical decision making, and more often hoped for and expected a different second opinion. Motives for second-opinion consultations differ greatly. Understanding the difference between internal and external motivation is necessary to develop strategies to prevent unnecessary second-opinion seeking. Additional studies are warranted to evaluate the objective and subjective outcomes of second-opinion consultations.


Preference for flavored wheat straw by lambs conditioned with intraruminal infusions of acetate and propionate.
We hypothesized that volatile fatty acids in rumen fluid are feedback signals that can condition food preferences or aversions in sheep. Three predictions were tested based on this hypothesis: 1) low doses of sodium propionate or sodium acetate condition preferences, but high doses condition aversions (Exp. 1 and 2); 2) preferences are not caused by osmotic load (Exp. 3 and 4); and 3) low doses of mixtures of acetate:propionate condition preferences (Exp. 4). In Exp. 1, 2, and 4, lambs were divided into four groups (10 lambs/group), and lambs in Exp. 3 were divided into two groups (five lambs/group). In all experiments, alfalfa pellets were the basal diet. On even days, half of the lambs were offered chopped wheat straw containing a distinctive flavor, whereas the other half received straw with a different flavor. During straw ingestion, different groups of lambs received intraruminal infusions of different concentrations (4, 8, or 12% of the daily DE received) of sodium propionate (Exp. 1), sodium acetate (Exp. 2), NaCl at osmotic loads equivalent to those when propionate supplied 4% of the daily DE received (Exp. 3), or different proportions of sodium acetate:sodium propionate (55:45 or 75:25% of the DE of the infusion [4% of the daily DE received]), or equimolar amounts of NaCl (Exp. 4). On odd days, the flavors were switched, and no infusions were administered. After 8 d of conditioning, lambs were offered a choice of wheat straw with the two distinctive flavors. Lambs preferred the flavor paired with the lowest doses of propionate (P = .07) and acetate (P = .08) but avoided the highest doses (P < .001). Excesses of VFA may condition aversions due to increases in rumen fluid osmolality and(or) excessive rates of supply of energy or sodium to the rumen. Lambs also preferred flavored straw associated with combinations of acetate and propionate (P < .001), especially at the highest concentration of propionate (P = .10). Lambs avoided NaCl in Exp. 3 (P < .001) and did not form preferences for NaCl in Exp. 4 (P > .05). Thus, osmolalities were not responsible for flavor preferences. In conclusion, our results support the hypothesis that food preferences and aversions reside along a continuum that depends on the amount of VFA infused.


[The short-latency reactions of the limbic cortex neurons to stimulation of the amygdaloid complex].
The extracellular electrical activities of the anterior limbic cortex (LC) neurons to electrical stimulation of amygdaloid central and basolateral nuclei in anesthetized cats was investigated. The orthodromic and antidromic responses of silent-neurons of LC were registered. These results show the existence of short and direct bilateral connections between the anterior LC and different amygdaloid nuclei. It is supposed, that these connections present a "quick-reacting" system, which takes part in the central regulation of the emotional responses.


Longitudinal analysis of acute-phase proteins in saliva in pig farms with different health status.
This study assesses the utility of saliva samples to monitor the time course of the acute-phase response to different viruses in pigs under field conditions by using time-resolved immunofluorometric assays (TR-IFMA). A total of 30 pigs from three different farms, located in Southeast Spain, were used. Farm 1 had outbreaks of porcine circovirus type 2, farm 2 had infections with porcine reproductive and respiratory syndrome virus and farm 3 had concomitant infections with both viruses. Serology was used to determine the time of seroconversion of pigs to two different pathogens. The levels of two acute-phase proteins (APPs), C-reactive protein (CRP) and haptoglobin (Hp), were measured in saliva and serum samples and compared with pig's serology. Kinetic curves of both APPs across the study obtained in saliva samples were similar to those of serum, with R of 0.68 and 0.78 for CRP and Hp, respectively. The median CRP and Hp concentrations in saliva were higher around the theorized time of infection, according to previous experimental studies, and at seroconversion of animals. CRP increments were apparent 1 week before the increments obtained in Hp. These findings indicate that salivary APP concentrations, by using TR-IFMA, can be used in longitudinal studies as non-invasive early indicators of health status.


The transcription factor Rpn4 activates its own transcription and induces efflux pump expression to confer fluconazole resistance in Candida auris.
Candida auris is a recently emerged pathogenic fungus of grave concern globally due to its resistance to conventional antifungals. This study takes a whole-genome approach to explore how C. auris overcomes growth inhibition imposed by the common antifungal drug fluconazole. We focused on gene disruptions caused by a "jumping genetic element" called transposon, leading to fluconazole resistance. We identified mutations in two genes, each encoding a component of the Ubr2/Mub1 ubiquitin-ligase complex, which marks the transcription regulator Rpn4 for degradation. When either protein is absent, stable Rpn4 accumulates in the cell. We found that Rpn4 activates the expression of itself as well as the main drug efflux pump gene CDR1 by binding to a PACE element in the promoter. Furthermore, we identified an amino acid change in Ubr2 in many resistant clinical isolates, contributing to Rpn4 stabilization and increased fluconazole resistance.


Nicotine dependence, abuse and craving: dimensionality in an Israeli sample.
Evidence-based changes planned for Diagnostic and Statistical Manual 5th edition (DSM-5) substance use disorders (SUDs) include combining dependence and three of the abuse criteria into one disorder and adding a criterion indicating craving. Because DSM-IV did not include a category for nicotine abuse, little empirical support is available for aligning the nicotine use disorder criteria with the DSM-5 criteria for other SUDs. Latent variable analyses, bootstrap tests and likelihood ratio tests were used to explore the unidimensionality, psychometric properties and information of the nicotine criteria. A sample of household residents selected from the Israeli population register yielded 727 life-time cigarette smokers. DSM-IV nicotine dependence criteria and proposed abuse and craving criteria, assessed with a structured interview. Three abuse criteria (hazardous use, social/interpersonal problems and neglect roles) were prevalent among smokers, formed a unidimensional latent trait with nicotine dependence criteria, were intermixed with dependence criteria across the severity spectrum and significantly increased the diagnostic information over the dependence-only model. A craving criterion was shown to fit well with the other criteria. Similar to findings from research on other substances, nicotine dependence, abuse and craving criteria appear to derive from a common underlying dimension. The results support alignment of nicotine criteria with those for alcohol and drug use disorders in Diagnostic and Statistical Manual 5th edition.


N-termini of fungal CSL transcription factors are disordered, enriched in regulatory motifs and inhibit DNA binding in fission yeast.
CSL (CBF1/RBP-Jκ/Suppressor of Hairless/LAG-1) transcription factors are the effector components of the Notch receptor signalling pathway, which is critical for metazoan development. The metazoan CSL proteins (class M) can also function in a Notch-independent manner. Recently, two novel classes of CSL proteins, designated F1 and F2, have been identified in fungi. The role of the fungal CSL proteins is unclear, because the Notch pathway is not present in fungi. In fission yeast, the Cbf11 and Cbf12 CSL paralogs play antagonistic roles in cell adhesion and the coordination of cell and nuclear division. Unusually long N-terminal extensions are typical for fungal and invertebrate CSL family members. In this study, we investigate the functional significance of these extended N-termini of CSL proteins. We identify 15 novel CSL family members from 7 fungal species and conduct bioinformatic analyses of a combined dataset containing 34 fungal and 11 metazoan CSL protein sequences. We show that the long, non-conserved N-terminal tails of fungal CSL proteins are likely disordered and enriched in phosphorylation sites and PEST motifs. In a case study of Cbf12 (class F2), we provide experimental evidence that the protein is proteolytically processed and that the N-terminus inhibits the Cbf12-dependent DNA binding activity in an electrophoretic mobility shift assay. This study provides insight into the characteristics of the long N-terminal tails of fungal CSL proteins that may be crucial for controlling DNA-binding and CSL function. We propose that the regulation of DNA binding by Cbf12 via its N-terminal region represents an important means by which fission yeast strikes a balance between the class F1 and class F2 paralog activities. This mode of regulation might be shared with other CSL-positive fungi, some of which are relevant to human disease and biotechnology.


Trait-specific consequences of inbreeding on adaptive phenotypic plasticity.
Environmental changes may stress organisms and stimulate an adaptive phenotypic response. Effects of inbreeding often interact with the environment and can decrease fitness of inbred individuals exposed to stress more so than that of outbred individuals. Such an interaction may stem from a reduced ability of inbred individuals to respond plastically to environmental stress; however, this hypothesis has rarely been tested. In this study, we mimicked the genetic constitution of natural inbred populations by rearing replicate Drosophila melanogaster populations for 25 generations at a reduced population size (10 individuals). The replicate inbred populations, as well as control populations reared at a population size of 500, were exposed to a benign developmental temperature and two developmental temperatures at the lower and upper margins of their viable range. Flies developed at the three temperatures were assessed for traits known to vary across temperatures, namely abdominal pigmentation, wing size, and wing shape. We found no significant difference in phenotypic plasticity in pigmentation or in wing size between inbred and control populations, but a significantly higher plasticity in wing shape across temperatures in inbred compared to control populations. Given that the norms of reaction for the noninbred control populations are adaptive, we conclude that a reduced ability to induce an adaptive phenotypic response to temperature changes is not a general consequence of inbreeding and thus not a general explanation of inbreeding-environment interaction effects on fitness components.


Association between Endocrine Disorders and Severe COVID-19 Disease in Pediatric Patients.
Though severe illness due to COVID-19 is uncommon in children, there is an urgent need to better determine the risk factors for disease severity in youth. This study aims to determine the impact a preexisting endocrine diagnosis has on severity of COVID-19 presentation in youth. The cross-sectional chart review study included all patients less than 25 years old with a positive SARS-CoV-2 PCR at St. Louis Children's Hospital between March 2020 and February 2021. Electronic medical record data for analysis included patient demographics, BMI percentile, inpatient hospitalization or admission to the pediatric intensive care unit (PICU), and the presence of a preexisting endocrine diagnosis such as diabetes mellitus (type 1 and type 2), adrenal insufficiency, and hypothyroidism. Two outcome measures were analyzed in multivariate analysis: inpatient admission and PICU admission. Adjusted odds ratios with a 95% CI were calculated using binary logistic regression, along with p values after Wald χ2 analysis. 390 patients were included in the study. Mean age was 123.1 (±82.2) months old. 50.3% of patients were hospitalized, and 12.1% of patients were admitted to intensive care. Preexisting diagnoses of diabetes mellitus, obesity, and hypothyroidism were associated with an increased risk of hospital and ICU admission, independent of socioeconomic status. This study provides evidence that unvaccinated youth with a preexisting diagnosis of obesity, hypothyroidism, or diabetes mellitus infected with COVID-19 are more likely to have a more severe clinical presentation requiring inpatient hospital admission and/or intensive care.


Optimal predictions in everyday cognition: the wisdom of individuals or crowds?
Griffiths and Tenenbaum (2006) asked individuals to make predictions about the duration or extent of everyday events (e.g., cake baking times), and reported that predictions were optimal, employing Bayesian inference based on veridical prior distributions. Although the predictions conformed strikingly to statistics of the world, they reflect averages over many individuals. On the conjecture that the accuracy of the group response is chiefly a consequence of aggregating across individuals, we constructed simple, heuristic approximations to the Bayesian model premised on the hypothesis that individuals have access merely to a sample of k instances drawn from the relevant distribution. The accuracy of the group response reported by Griffiths and Tenenbaum could be accounted for by supposing that individuals each utilize only two instances. Moreover, the variability of the group data is more consistent with this small-sample hypothesis than with the hypothesis that people utilize veridical or nearly veridical representations of the underlying prior distributions. Our analyses lead to a qualitatively different view of how individuals reason from past experience than the view espoused by Griffiths and Tenenbaum.


Tea consumption and the reduced risk of colon cancer -- results from a national prospective cohort study.
This study examines the relationship between tea consumption and colon cancer risk in the US population. Data from the NHANES I Epidemiologic Follow-up study (NHEFS) were used to examine the hypothesis. Cox proportional hazard models were used to examine the hypothesis of a protective effect of frequent tea consumption on colon cancer occurrence. Due to differences in the precision of the exposure data, we analysed two cohort periods based on the NHEFS. Cohort I was based on the survey conducted at the NHEFS baseline and Cohort II began at the first follow-up. After excluding non-incidence cases and cases lost to follow-ups, there were 2359 tea users and 6498 non-tea users at baseline and 7656 tea users and 4514 non-tea users at the first follow-up. : After adjusting for confounders, the relative risks of colon cancer are 0.57 (95% confidence interval (CI) 0.42, 0.78) and 0.59 (95% 1.00) for subjects who consumed <or=1.5 cups and >1.5 cups per day, respectively, compared with non-tea users in Cohort II. Although more women consumed tea and the mean intake was higher, the preventive effect of tea consumption on colon cancer was found predominantly in men. The relative risks of colon cancer are 0.41 (95% 0.66) for men who consumed <or=1.5 cups day-1 and 0.30 (95% 0.98) for >1.5 cups day-1 of tea consumption (P-value for trend <0.01). No significant results were found in Cohort I. This study suggests an inverse association between colon cancer risk and habitual tea consumption.


Covalent layer-by-layer thin films with charge-transfer chromophores: side chain engineering for efficient Ag+ ion recognition in aqueous solutions.
Covalent layer-by-layer (LbL) thin films are fabricated by a highly efficient [2+2] cycloaddition-retroelectrocyclization (CA-RE) reaction of aniline-substituted alkyne and 7,7,8,8-tetracyanoquinodimethane (TCNQ) moieties. Polystyrene bearing aniline-substituted alkyne side chains and TCNQ polyester were prepared as precursor polymers, then sequentially deposited and fixed by covalent bonds on an indium-tin-oxide (ITO) substrate. The successful alternate growth of the films was demonstrated by many techniques including absorption spectroscopy, surface roughness, and redox activities. Interestingly, the water contact angles of the film surface were dependent on the side chains of the polystyrene derivatives. When hydrophilic triethylene glycol was employed, the surface hydrophilicity was similar to that of the TCNQ polyester. In contrast, the use of hydrophobic n-hexyl groups resulted in a significant difference in the water contact angles between the polystyrene and TCNQ polyester. A similar difference occurred when the Ag+ ion recognition was studied by soaking the LbL films in aqueous solutions. The LbL films constructed from the polystyrene with triethylene glycol chains displayed a more rapid and significant recognition of Ag+ ions than those constructed from the polystyrene with n-hexyl chains.


[Theoretical evidence for the need to suppress parasitic recirculation in the futile cycle fructose-6-P--fructose-1,6-P2].
In connection with the discussion of a possible role of the futile cycle fructose-6-P in equilibrium or formed from fructose-1,6,-P2 in the regulation of the carbohydrate energy metabolism, simple stoichiometric models of glycolysis and gluconeogenesis are examined which take account of substrate recirculation in the cycle. The recirculation is shown to involve wasteful expenditure of ATP and to divert fructose-1,6-P2 from glycolytic phosphorylation ADP and fructose-6-P from glucose or glycogen resynthesis. As a result, the glycolytic system loses its capacity to stabilize the level of ATP, while gluconeogenesis proves to be not efficient. From the analysis performed it is concluded that normal energy metabolism should involve allosteric regulatory mechanisms to suppress substrate recirculation in the cycle.


A robust tolerance design method based on process capability.
This paper presents a method for robust tolerance design in terms of Process Capability Indices (PCI). The component tolerance and the suitable manufacturing processes can be selected based on the real manufacturing context. The robustness of design feasibility under the effect of uncertainties is also discussed. A comparison between the results obtained by the proposed model and other methods indicates that robust and reliable tolerance can be obtained.


Supine Percutaneous Nephrolithotomy in the Galdako-Modified Valdivia Position: A High-Volume Single Center Experience.
To describe the technique and outcomes of supine percutaneous nephrolithotomy (PCNL) in the Galdako-modified Valdivia position. Prospective data were collected from 303 patients undergoing PCNL in the Galdako-modified Valdivia position at our institution between 2007 and 2015. We report our technique with outcomes of operative times, stone-free rate (SFR), and complications. A total of 202 solitary stones with a mean size of 17.19 ± 5.82 mm, 42 stones in multiple calices, and 57 staghorns were treated. Mean operative time was 79.79 ± 35.72 minutes. A total of 244/303 (80.5%) patients had clearance or <2 mm stone on postoperative CT. Minor complications (Clavien-Dindo Classification [CDC] 1-2) occurred in 59 (19.5%) cases and major complications (CDC ≥3) occurred in 22 (7.3%) cases. Five (1.7%) cases required postoperative blood transfusion for bleeding complications. We demonstrate that PCNL in a high-volume center to be safe and efficacious in the Galdalko-modified Valdivia position. We show equivalent SFR and complication rates to large studies of prone PCNL, with potential to decrease operative time. In particular, supine position is optimum for obese or high-risk patients with complex stone disease, because of ease of positioning and lower theoretical risk of anesthetic complications.


Cold Agglutinin Disease in a Rhesus Macaque (Macaca mulatta).
Cold agglutinin disease (CAD) is a condition involving anemia and its related symptoms; it is caused by autoantibodies that bind and agglutinate red blood cells in areas susceptible to hypothermia, such as extremities exposed to cold temperatures. CAD is rare, with 5 to 20 human cases per million individuals. In this report, we describe a case of CAD in a previously healthy and experimentally naïve adult Indian rhesus macaque that was housed indoors and presented with blood in the urine. After our observations of hemoglobinuria and anemia led us to suspect CAD, we demonstrated that the macaque's blood agglutinated at reduced temperatures. We also noticed that the provision of cold foraging treats triggered episodes of hemoglobinuria. Further investigation revealed that serum from the macaque agglutinated RBCs in vitro with high thermal amplitude (at or below 30 °C) and had an antibody titer of 8 to 32. The serum contained autoantibodies of the immunoglobulin M (IgM) isotype; agglutinins of the IgG isotype were not detected. The cold-dependent IgM autoantibodies in the serum from the affected macaque reacted against a common RBC antigen because RBCs collected from other macaques were bound and agglutinated by the affected animal's IgM under cold conditions. This in vitro binding activity was reversible when the test temperature was returned to normal body temperature (37 °C). These findings demonstrated cold-dependent RBC-specific IgM agglutinins and led us to a diagnosis of CAD. This is the first documented case of spontaneous CAD in a rhesus macaque.


Functional expression of rat cytochrome c in Saccharomyces cerevisiae.
To determine whether a mammalian cytochrome c could efficiently replace iso-1-cytochrome c, which is encoded by the yeast CYC1 gene, the coding sequence of RC9 (a nondefective processed gene from rat) was cloned in both single- and multiple-copy expression vectors under the direction of the yeast alcohol dehydrogenase 1 (ADC1) promoter. Upon transformation of a CYC1 deletion strain, the multiple-copy construct restored a wild-type growth rate on lactate medium; such growth normally requires derepressed amounts of iso-1-cytochrome c. These transformants expressed a level of hybrid ADC1/RC9 mRNA approximately 5- to 10-fold greater than the amount of message from the endogenous ADC1 gene and produced a steady-state level of rat cytochrome c equivalent to that of the wild-type yeast protein. A requirement for the vector was evidenced by its absence in all transformants that lost the lactate growth phenotype after propagation in nonselective medium. In contrast, the level of vector-specific message in single copy was equivalent to that of the endogenous ADC1 mRNA, but transformants exhibited no significant growth on lactate. Constructions having a small deletion or a mammalian intron within the rat cytochrome c coding region failed to support lactate-dependent growth, indicating that complementation depends upon proper translation of the correct rat coding sequence. Therefore, the rat polypeptide, when expressed at normal physiological levels, is recognized by the yeast machinery involved in the multiple steps required for the processing and transport of an active cytochrome c as well as its functional interaction with the respiratory apparatus.


Spiral cleavages determine the left-right body plan by regulating Nodal pathway in monomorphic gastropods, Physa acuta.
The handedness of gastropods is genetically determined, but the molecular nature of the gene responsible and the associated mechanisms remain unknown. In order to characterize the chiromorphogenesis pathway starting from the gene to the left-right asymmetric body plan, we have closely analyzed the cytoskeletal dynamics of the Physa (P.) acuta embryo, a fresh water non-dimorphic sinistral snail, during the early developmental stage by mechanically altering the handedness of the embryos at the critical spiral third cleavage. A fertile situs inversus was created and the nodal-Pitx gene expression patterns were completely mirror imaged to the wild type at the trochophore stage. Together with our previous work on Lymnaea (L.) stagnalis, we could show that chirality is established at the third cleavage, as dictated by the single handedness-determining gene locus, and then chirality information is transferred via subsequent spiral fourth and fifth cleavages to the later developmental stage, dictating the nodal-Pitx expression pathway. The cytoskeletal dynamics of manipulated and non-manipulated embryos of sinistral P. acuta and dextral dominant L. stagnalis are compared.


Synaptic number changes in the medial prefrontal cortex across adolescence in male and female rats: A role for pubertal onset.
Adolescence is a unique period of development, marked by maturation of the prefrontal cortex (PFC), a region important for executive functioning. During this time, the human PFC decreases in overall volume and thickness. Likewise in adolescent rodents, losses of neurons, dendrites, dendritic spines and neurotransmitter receptors have been documented within the medial prefrontal cortex (mPFC), sometimes with sex and layer specificity. However, changes in the number of synapses during this time have not been examined. In the present study, we stereologically quantified the number of synaptophysin-immunoreactive boutons in the male and female rat mPFC across multiple time points from the juvenile period through adulthood (postnatal days (P) 25, 35, 45, 60 and 90). In females, there was a significant decrease in synaptophysin boutons between P35 and P45, coinciding with the onset of puberty. In males, there was no significant main effect of age on synaptophysin boutons; however, in both males and females, pubertal onset was associated with significant synaptic losses. These results suggest that puberty is a critical period for synaptic pruning within the rat mPFC, potentially contributing to maturation of adolescent executive function. Synapse 70:361-368, 2016. © 2016 Wiley Periodicals, Inc.


Estimating the global distribution of field size using crowdsourcing.
There is an increasing evidence that smallholder farms contribute substantially to food production globally, yet spatially explicit data on agricultural field sizes are currently lacking. Automated field size delineation using remote sensing or the estimation of average farm size at subnational level using census data are two approaches that have been used. However, both have limitations, for example, automatic field size delineation using remote sensing has not yet been implemented at a global scale while the spatial resolution is very coarse when using census data. This paper demonstrates a unique approach to quantifying and mapping agricultural field size globally using crowdsourcing. A campaign was run in June 2017, where participants were asked to visually interpret very high resolution satellite imagery from Google Maps and Bing using the Geo-Wiki application. During the campaign, participants collected field size data for 130 K unique locations around the globe. Using this sample, we have produced the most accurate global field size map to date and estimated the percentage of different field sizes, ranging from very small to very large, in agricultural areas at global, continental, and national levels. The results show that smallholder farms occupy up to 40% of agricultural areas globally, which means that, potentially, there are many more smallholder farms in comparison with the two different current global estimates of 12% and 24%. The global field size map and the crowdsourced data set are openly available and can be used for integrated assessment modeling, comparative studies of agricultural dynamics across different contexts, for training and validation of remote sensing field size delineation, and potential contributions to the Sustainable Development Goal of Ending hunger, achieve food security and improved nutrition and promote sustainable agriculture.


[Mechanism of growth limitations in a recombinant strain of Escherichia coli].
A recombinant Escherichia coli strain with the cloned gene of restriction endonuclease EcoR-V was studied. The diauxic growth of this strain under batch conditions, the composition of excreted products and the hyperbolic shape of the chemostat growth curve has made it possible to formulate the following hypothesis. At a high flow rate, one of the first reactions in the tricarboxylic acid cycle or a reaction directly preceding the cycle is a limiting step in the metabolism of E. coli cells. The saturation of the limiting reaction with a substrate at a flow rate higher than the critical one may be responsible for the hyperbolic profile of chemostat curves. The above hypothesis was confirmed by experiments conducted under extreme conditions, in which the composition of the growth medium was changed and the temperature was raised so that the cloned protein synthesis was depressed. Under such conditions, the enzyme composition of the cell was fixed and determined by the physiological state of the culture at a time when the temperature was elevated. The hypothesis was also supported by the results obtained in growing the strain with the induced protein synthesis in media to which various compounds were added.


Anti-inflammatory role of cilostazol in vascular smooth muscle cells in vitro and in vivo.
Cilostazol is a selective inhibitor of phosphodiesterase 3, by which it increases intracellular cAMP and activates protein kinase A, thereby inhibiting platelet aggregation and inducing peripheral vasodilation. We investigated whether cilostazol might prevent nuclear factor (NF)-kappaB activation by activating AMP-activated protein kinase (AMPK) in vascular smooth muscle cells (VSMC). Cilostazol was observed to activate AMPK, as well as its downstream target, acetyl-CoA carboxylase, in rat VSMC. Phosphorylation of AMPK with cilostazol was not affected by co-treatment with an adenylate cyclase inhibitor, SQ 22536. Furthermore, a cell-permeable cyclic AMP analog, pCTP-cAMP, did not influence cilostazol-induced AMPK phosphorylation. These findings suggest that cilostazol-induced AMPK activation occurs through a signalling pathway independent of cyclic AMP. Cilostazol dose-dependently inhibited LPS-induced NF-kappaB activation in the present study. It was also observed to inhibit LPS-induced iNOS gene promoter activity and iNOS gene expression, resulting in markedly reduced NO production. An AMPK inhibitor compound C or siRNA for AMPK attenuated the observed cilostazol-induced inhibition of NF-kappaB activation by LPS. Ingestion of cilostazol inhibited NF-kappaB activation, as well as the induction of iNOS mRNA and protein expression, within the aortas of LPS-treated rats. In light of these findings, we suggest that cilostazol might attenuate cytokine-induced expression of the iNOS gene by inhibiting NF-kappaB following AMPK activation in VSMC.


Retinal projection or size constancy as determinants of the horizontal-vertical illusion?
To compare the influence of the projected retinal size and of the figure size on the perception of the horizontal-vertical illusion, the target size, the viewing distance, and the slant of an illusion figure were varied. In the first experiment the illusion produced by two figures of the same object size but of different retinal size was compared with that of two figures projecting the same retinal size but differing in object size. The illusion diminished when the size of the retinal projection was increased, whereas a change in figure size did not change the illusion. In Exp. II the illusion figure was tilted backwards which reduced the retinal projection of the 'vertical' figure limb. The illusion decreased and became negative as a function of the retinal projection, but this decrease was relatively small compared with the reduction of the retinal image. The results are interpreted as supporting a retinal origin as an explanation of the illusion. Although there is strong evidence for size-constancy scaling in a tilted figure, constancy scaling is considered of minor importance as a determinant of the usual illusion.


Respiratory physiology and energy conservation efficiency of Campylobacter jejuni.
A study of the electron transport chain of the human intestinal pathogen Campylobacter jejuni revealed a rich complement of b- and c-type cytochromes. Two c-type cytochromes were partially purified: one, possibly an oxidase, bound carbon monoxide whereas the other, of high potential was unreactive with carbon monoxide. Respiratory activities determined with membrane vesicles were 50- to 100-fold higher with formate and hydrogen than with succinate, lactate, malate, or NADH as substrates. Evidence for three terminal respiratory components was obtained from respiratory kinetic studies employing cyanide, and the following Ki values for cyanide were determined from Dixon plots: ascorbate + reduced N,N,N', N'-tetramethyl-p-phenylenediamine, K1 + 3.5 muM; malate, K1 = 55 muM; and hydrogen, K1 = 4.5 muM. Two oxidases (K1 = 90 muM, 4.5 mM) participated in the oxidation of succinate, lactate, and formate. Except with formate, 37 muM HQNO inhibited respiration by approximately 50%. Carbon monoxide had little inhibitory effect on respiration except under low oxygen tension (less than 10% air saturation). The stoichiometry of respiratory-driven proton translocation (H+/O) determined with whole cells was approximately 2 for all substrates examined except hydrogen (H+/) = 3.7) and formate (H+/O = 2.5). The higher stoichiometries observed with hydrogen and formate are consistent with their respective dehydrogenase being located on the periplasmic face of the cytoplasmic membrane. The results of this study suggest that the oxidation of hydrogen and formate probably serves as the major sources of energy for growth.


Topology: a novel method to describe branching patterns in Peronospora viciae colonies.
Topology provides a novel means to describe branching patterns and has not been applied to fungal colonies previously. For any branched structure, various topologies are possible, and these lie between two extremes, a herringbone pattern (main axis with primary laterals) and a dichotomous pattern (highly branched system). We applied topological methods to colonies of Peronospora viciae 48 h after inoculation of Pisum sativum leaves. The methods are based on two simulations, one developed for channel networks such as found in river systems and another for biological systems. Although not a true herringbone form, the Peronospora viciae colonies have a strong herringbone element within their growth pattern. All 25 colonies analysed fell into the random distribution according to the confidence limits calculated from simulations for biological systems. These confidence limits, however, represent the percentile distribution of all simulated networks, and only those structures with a perfect herringbone or dichotomous topology fall outside the range. The tendency of P. viciae colonies towards herringbone growth is reflected by the topological indices for altitude and external pathlength (a(obs)/E(a) and pe(obs)/E(pe), where a = altitude, pe = external pathlength, obs = observed for the P. viciae colonies and E = expected values for random growth), and the slope of the regression analysis for a(obs) and pe(obs). We consider this trend as significant because it was consistent for all but one of the colonies, and implies that growth can be envisaged as an intermediate between random and herringbone topology. It is proposed that initial herringbone growth may reflect a strategy that is aimed at overcoming host resistance, achieving rapid colonisation of infected tissue and maximising the potential for nutrient acquisition. This topology would also increase the likelihood of finding a compatible mating type for reproduction between heterothallic isolates.


Generation of an environmental niche for neural stem cell development by the extracellular matrix molecule tenascin C.
Stem cells in the embryonic mammalian CNS are initially responsive to fibroblast growth factor 2 (FGF2). They then undergo a developmental programme in which they acquire epidermal growth factor (EGF) responsiveness, switch from the production of neuronal to glial precursors and become localized in specialized germinal zones such as the subventricular zone (SVZ). Here we show that extracellular matrix molecules act as regulators of this programme. Tenascin C is highly expressed in the SVZ, and transgenic mice lacking tenascin C show delayed acquisition of the EGF receptor. This results from alterations in the response of the stem cells to the growth factors FGF2 and bone morphogenic protein 4 (BMP4), which normally promote and inhibit acquisition of the EGF receptor, respectively. Tenascin C-deficient mice also have altered numbers of CNS stem cells and these stem cells have an increased probability of generating neurones when grown in cell culture. We conclude that tenascin C contributes to the generation of a stem cell 'niche' within the SVZ, acting to orchestrate growth factor signalling so as to accelerate neural stem cell development.


The Pseudoalteromonas luteoviolacea L-amino Acid Oxidase with Antimicrobial Activity Is a Flavoenzyme.
The marine environment is a rich source of antimicrobial compounds with promising pharmaceutical and biotechnological applications. The Pseudoalteromonas genus harbors one of the highest proportions of bacterial species producing antimicrobial molecules. For decades, the presence of proteins with L-amino acid oxidase (LAAO) and antimicrobial activity in Pseudoalteromonas luteoviolacea has been known. Here, we present for the first time the identification, cloning, characterization and phylogenetic analysis of Pl-LAAO, the enzyme responsible for both LAAO and antimicrobial activity in P. luteoviolacea strain CPMOR-2. Pl-LAAO is a flavoprotein of a broad substrate range, in which the hydrogen peroxide generated in the LAAO reaction is responsible for the antimicrobial activity. So far, no protein with a sequence similarity to Pl-LAAO has been cloned or characterized, with this being the first report on a flavin adenine dinucleotide (FAD)-containing LAAO with antimicrobial activity from a marine microorganism. Our results revealed that 20.4% of the sequenced Pseudoalteromonas strains (specifically, 66.6% of P. luteoviolacea strains) contain Pl-laao similar genes, which constitutes a well-defined phylogenetic group. In summary, this work provides insights into the biological significance of antimicrobial LAAOs in the Pseudoalteromonas genus and shows an effective approach for the detection of novel LAAOs, whose study may be useful for biotechnological applications.


Investigations on the Electrochemical Atomic Layer Growth of Bi₂Se₃ and the Surface Limited Deposition of Bismuth at the Silver Electrode.
The Electrochemical Atomic Layer Deposition (E-ALD) technique is used for the deposition of ultrathin films of bismuth (Bi) compounds. Exploiting the E-ALD, it was possible to obtain highly controlled nanostructured depositions as needed, for the application of these materials for novel electronics (topological insulators), thermoelectrics and opto-electronics applications. Electrochemical studies have been conducted to determine the Underpotential Deposition (UPD) of Bi on selenium (Se) to obtain the Bi₂Se₃ compound on the Ag (111) electrode. Verifying the composition with X-ray Photoelectron Spectroscopy (XPS) showed that, after the first monolayer, the deposition of Se stopped. Thicker deposits were synthesized exploiting a time-controlled deposition of massive Se. We then investigated the optimal conditions to deposit a single monolayer of metallic Bi directly on the Ag.


Characterisation of isolates of Haemophilus paragallinarum from Indonesia.
To characterise 18 isolates of Haemophilus paragallinarum isolated from chickens in Indonesia. The isolates were identified to species level by traditional phenotypic methods. Six of the isolates were also identified by a species-specific polymerase chain reaction. Fourteen of the isolates were examined for resistance to a panel of seven antimicrobial agents using a disc diffusion method. All 18 isolates were serotyped according to the Page scheme using reference antisera in a haemagglutination inhibition test. Four of the 18 isolates were obtained from indigenous (kampung) chickens, with the remainder being from typical intensive poultry production systems. The 18 isolates were obtained from 11 outbreaks that showed the typical clinical signs of infectious coryza and 11 of the isolates were obtained from chickens that had been vaccinated with infectious coryza vaccines. All 18 isolates were confirmed as H paragallinarum by biochemical testing and six isolates were also identified as H paragallinarum by the polymerase chain reaction test. Eleven isolates were resistant to erythromycin and streptomycin, 10 to neomycin, eight to oxytetracycline, five isolates to doxycycline, three to sulphamethoxazoltrimethoprim but only one to ampicillin. Seven isolates were Page serovar A, four were Page serovar B and seven were Page serovar C. The presence of all three Page serovars (A, B and C) has been confirmed for the first time in Indonesian chickens. As the majority of the infectious coryza vaccines in use in Indonesia contain only serovar A and C, the presence of serovar B in chickens indicates that the protection by these bivalent vaccines would be reduced. The use of trivalent infectious coryza vaccines that contain serovars A, B and C is recommended for use in Indonesia.


Multiple sclerosis reduces sensitivity to immediate reward during decision making.
Individuals with multiple sclerosis (MS) often face important health-related and financial decisions that involve trade-offs between short-term and long-term benefits, yet decision making is rarely studied in MS patients. The temporal discounting paradigm is a useful tool for investigating such time-dependent choices in humans. Here, we investigated whether patients with relapsing-remitting MS differed from healthy controls when making choices between hypothetical monetary rewards available at different points in time. Participants were tested in two conditions: in one, the choice was between a smaller amount of money available immediately and a larger amount of money available at a later date; in the other, a fixed delay of 60 days was added to both options. We found that, compared with healthy controls, MS patients favored less the sooner reward in the condition involving an immediate reward, whereas no difference between MS patients and the control group emerged in the condition involving only delayed rewards. Moreover, the decreased immediacy bias was corroborated by lower scores at scale that assesses responsiveness to rewards in MS patients. Taken together, these findings indicate reduced sensitivity to immediate reward and a consequent stronger willingness to defer gratification in MS individuals. (PsycINFO Database Record


Racial disparities among Asian Americans with atrial fibrillation: An analysis from the NCDR® PINNACLE Registry.
There is paucity of data on Atrial Fibrillation (AF) management and associated clinical outcomes among Asian Americans. This study sought to investigate baseline risk factor profiles, racial disparities in clinical management and adverse clinical outcomes among White and Asian Americans. We used National Cardiovascular Data Registry (NCDR®) Practice Innovation and Clinical Excellence (PINNACLE) registry and linked Centers of Medicare and Medicaid Services data to identify Asian and White patients with AF between January 1, 2013-June 30, 2018. We compared rates of baseline risk factors, management strategies (rate versus rhythm control), anticoagulation use and rates of adverse events between racial groups. The two race groups were compared using hierarchical multivariable adjusted regression models to account for site and confounders. In total, 1,359,827 patients (18,793 Asians and 1,341,034 Whites) were included in our analysis. Compared to White Americans, Asian Americans were more likely to use a rate control strategy (Odds Ratio [OR]: 1.20, 95% Confidence Interval [CI]: 1.15-1.25) and lower odds of rhythm control strategy (atrial ablations, cardioversions, or use of antiarrhythmic drugs) (OR: 0.83, 95% CI: 0.80-0.87) in adjusted analysis. Use of oral anticoagulants and direct oral anticoagulants were similar. There were no significant race-based differences in likelihood of all-cause mortality, stroke, and bleeding requiring hospitalization. Analyses performed using propensity score matching were consistent with the main results. Asian Americans with AF have a lower likelihood of being managed with rhythm control strategies. Overall use of OAC and AF related adverse events remain similar between the two racial groups.


Strain Differences in Presynaptic Function: PROTEOMICS, ULTRASTRUCTURE, AND PHYSIOLOGY OF HIPPOCAMPAL SYNAPSES IN DBA/2J AND C57Bl/6J MICE.
The inbred strains C57BL/6J and DBA/2J (DBA) display striking differences in a number of behavioral tasks depending on hippocampal function, such as contextual memory. Historically, this has been explained through differences in postsynaptic protein expression underlying synaptic transmission and plasticity. We measured the synaptic hippocampal protein content (iTRAQ (Isobaric Tags for Relative and Absolute Quantitation) and mass spectrometry), CA1 synapse ultrastructural morphology, and synaptic functioning in adult C57BL/6J and DBA mice. DBA mice showed a prominent decrease in the Ras-GAP calcium-sensing protein RASAL1. Furthermore, expression of several presynaptic markers involved in exocytosis, such as syntaxin (Stx1b), Ras-related proteins (Rab3a/c), and rabphilin (Rph3a), was reduced. Ultrastructural analysis of CA1 hippocampal synapses showed a significantly lower number of synaptic vesicles and presynaptic cluster size in DBA mice, without changes in postsynaptic density or active zone. In line with this compromised presynaptic morphological and molecular phenotype in DBA mice, we found significantly lower paired-pulse facilitation and enhanced short term depression of glutamatergic synapses, indicating a difference in transmitter release and/or refilling mechanisms. Taken together, our data suggest that in addition to strain-specific postsynaptic differences, the change in dynamic properties of presynaptic transmitter release may underlie compromised synaptic processing related to cognitive functioning in DBA mice.


The putative tumor suppressor gene EphA3 fails to demonstrate a crucial role in murine lung tumorigenesis or morphogenesis.
Treatment of non-small cell lung cancer (NSCLC) is based on histological analysis and molecular profiling of targetable driver oncogenes. Therapeutic responses are further defined by the landscape of passenger mutations, or loss of tumor suppressor genes. We report here a thorough study to address the physiological role of the putative lung cancer tumor suppressor EPH receptor A3 (EPHA3), a gene that is frequently mutated in human lung adenocarcinomas. Our data shows that homozygous or heterozygous loss of EphA3 does not alter the progression of murine adenocarcinomas that result from Kras mutation or loss of Trp53, and we detected negligible postnatal expression of EphA3 in adult wild-type lungs. Yet, EphA3 was expressed in the distal mesenchyme of developing mouse lungs, neighboring the epithelial expression of its Efna1 ligand; this is consistent with the known roles of EPH receptors in embryonic development. However, the partial loss of EphA3 leads only to subtle changes in epithelial Nkx2-1, endothelial Cd31 and mesenchymal Fgf10 RNA expression levels, and no macroscopic phenotypic effects on lung epithelial branching, mesenchymal cell proliferation, or abundance and localization of CD31-positive endothelia. The lack of a discernible lung phenotype in EphA3-null mice might indicate lack of an overt role for EPHA3 in the murine lung, or imply functional redundancy between EPHA receptors. Our study shows how biological complexity can challenge in vivo functional validation of mutations identified in sequencing efforts, and provides an incentive for the design of knock-in or conditional models to assign the role of EPHA3 mutation during lung tumorigenesis.


Treatment of hyperlipidaemia with fenofibrate and related fibrates.
Fenofibrate is the most widely used fibrate. Its efficacy and tolerability in the treatment of hypertriglyceridaemia and combined hyperlipidaemia have been demonstrated in several clinical trials. To review the pharmacology, lipid-lowering and extra-lipid effects of fenofibrate and to preview ABT-335, an investigational new fenofibric acid molecule. The effects of fenofibrate are mediated through the active metabolite fenofibric acid, and are described in detail in the paper. ABT-335 is a salt of fenofibric acid and, unlike fenofibrate, does not require first pass metabolism to the active moiety. ABT-335 is being developed for combination use with statins, and has recently completed three large Phase III randomised controlled trials in which the efficacy and safety of ABT-335 in combination with the three most commonly prescribed statins, atorvastatin, simvastatin and rosuvastatin, was evaluated in patients with mixed dyslipidaemia. ABT-335 in combination with statins may provide a safe and efficacious treatment modality that enables achievement of several therapeutic goals in patients with mixed dyslipidaemia who have high cardiovascular risk.


Molecular characterization of the sodium channel subunits expressed in mammalian cerebellar Purkinje cells.
Inactivating and noninactivating Na+ conductances are known to generate, respectively, the rising phase and the prolonged plateau phase of cerebellar Purkinje cell (PC) action potentials. These conductances have different voltage activation levels, suggesting the possibility that two distinct types of ion channels are involved. Single Purkinje cell reverse transcription-PCR from guinea pig cerebellar slices identified two Na+ channel alpha subunit transcripts, the orthologs of RBI (rat brain I) and Nach6/Scn8a. The latter we shall name CerIII. In situ hybridization histochemistry in rat brain demonstrated broad CerIII expression at high levels in many neuronal groups in the brain and spinal cord, with little if any expression in white matter, or nerve tracts. RBII (rat brain II), the most commonly studied recombinant Na+ channel alpha subunit is not expressed in PCs. As the absence of Scn8a has been correlated with motor endplate disease (med), in which transient sodium currents are spared, RBI appears to be responsible for the transient sodium current in PC. Conversely, jolting mice with a mutated Scn8a message demonstrates PC abnormalities in rapid, simple spike generation, linking CerIII to the persistent sodium current.


Predicting cellulose solvating capabilities of acid-base conjugate ionic liquids.
Different acid-base conjugates were made by combining a range of bases and superbases with acetic and propionic acid. Only the combinations that contained superbases were capable of dissolving cellulose. Proton affinities were calculated for the bases. A range, within which cellulose dissolution occurred, when combined with acetic or propionic acid, was defined for further use. This was above a proton affinity value of about 240 kcal mol(-1) at the MP2/6-311+G(d,p)//MP2/ 6-311+G(d,p) ab initio level. Understanding dissolution allowed us to determine that cation acidity contributed considerably to the ability of ionic liquids to dissolve cellulose and not just the basicity of the anion. By XRD analyses of suitable crystals, hydrogen bonding interactions between anion and cation were found to be the dominant interactions in the crystalline state. From determination of viscosities of these conjugates over a temperature range, certain structures were found to have as low a viscosity as 1-ethyl-3-methylimidazolium acetate, which was reflected in their high rate of cellulose dissolution but not necessarily the quantitative solubility of cellulose in those ionic liquids. 1,5-Diazabicyclo[4.3.0]non-5-enium propionate, which is one of the best structures for cellulose dissolution, was then distilled using laboratory equipment to demonstrate its recyclability.


21st-century rise in anthropogenic nitrogen deposition on a remote coral reef.
With the rapid rise in pollution-associated nitrogen inputs to the western Pacific, it has been suggested that even the open ocean has been affected. In a coral core from Dongsha Atoll, a remote coral reef ecosystem, we observe a decline in the 15N/14N of coral skeleton-bound organic matter, which signals increased deposition of anthropogenic atmospheric N on the open ocean and its incorporation into plankton and, in turn, the atoll corals. The first clear change occurred just before 2000 CE, decades later than predicted by other work. The amplitude of change suggests that, by 2010, anthropogenic atmospheric N deposition represented 20 ± 5% of the annual N input to the surface ocean in this region, which appears to be at the lower end of other estimates.


Two new species of Hypostomus suckermouth-armoured catfishes (Teleostei: Loricariidae) from central Brazil.
This study describes two new endemic Hypostomus species from central Brazil, which were previously identified as genetically distinct lineages in a recent genomic study that recommended their testing and potential description based on morphological data. A machine learning classification procedure (random forest) was used to investigate morphological variation and identify putatively diagnostic characters for these candidate species and revealed that each is morphologically distinct. The new species Hypostomus cafuringa is characterized by small size, dark spots under a light background, deeper caudal peduncle and shorter first ray of the pectoral fin and base of the dorsal fin when compared to congeneric species from the region. H. cafuringa is known from the headwaters of the Maranhão River, upper Tocantins River basin, Distrito Federal, Brazil. The second new species, Hypostomus crulsi, is characterized by dark spots under a light background, absence of plates along the abdomen region, shorter first ray of the pelvic fin, shorter first ray of the pectoral fin and smaller body size. H. crulsi is known from the headwaters of the São Bartolomeu River, upper Paraná River basin, Distrito Federal, Brazil. The rapid conversion of natural habitats for agricultural development and the isolation of protected areas represent a serious threat to the continued existence of these two newly described endemic species, which warrant conservation assessment.


Modulation of murine Peyer's patch immunoglobulin A response by enterotoxigenic Escherichia coli.
Enterotoxigenic Escherichia coli (ETEC) may have profound effects on the capacity of gut-associated lymphoid tissue to mount a secretory immune response because of the potential ability of heat-stable toxin or heat-labile toxin to modulate the immune response. To examine the effects of ETEC or its purified enterotoxins upon the humoral immune response of murine small intestinal Peyer's patch lymphocytes, BDF1 (lipopolysaccharide-responder) and C3H/HeJ (lipopolysaccharide-nonresponder) mice were orally primed with sheep erythrocytes (SRBC) four times during a 2-week period to initiate differentiation of Peyer's patch B lymphocytes to cells committed to anti-SRBC immunoglobulin A (IgA) production. Halfway through the oral priming regimen the mice were gastrically intubated with 10(8) ETEC, 10(8) non-ETEC, or saline. ETEC persisted in the small intestine for at least 7 days at a level of 10(3) to 10(4) bacteria per mouse. Seven days after the last oral dosing with SRBC, Peyer's patch lymphocytes were removed from infected or saline-treated mice and incubated in vitro with SRBC. The ETEC infection had a small effect on the anti-SRBC IgM plaque-forming cell response of SRBC-primed mice but inhibited significantly the anti-SRBC IgA plaque-forming cell response in both BDF1 and C3H/HeJ mice as compared with uninfected controls. The non-ETEC, an isolate from normal mouse small intestine, had no significant effect on either IgM or IgA anti-SRBC plaque-forming cell response. Purified heat-labile toxin, not heat-stable toxin, alone in a dose-dependent manner significantly inhibited both the IgA and IgM plaque-forming cell response of Peyer's patch lymphocytes from primed mice. These data suggest that ETEC can inhibit the development of the gut-associated lymphoid tissue IgA immune response through the immunopharmacological effect of an enterotoxin, the heat-labile toxin.


Gene expression patterns in hypoxic and post-hypoxic adult rat retina with special reference to the NMDA receptor and its interactome.
A gene expression analysis of hypoxic rat retina was undertaken to gain a deeper understanding of the possible molecular mechanisms that underlie hypoxia-induced retinal pathologies and identify possible therapeutic targets. Rats were made severely hypoxic (6%-7% O(2)) for 3 h. Some rats were sacrificed at this time, and others were allowed to recover for 24 h under normoxic conditions. A focused oligonucleotide microarray of 1,178 genes, qRT-PCR of selected transcripts, and western analysis of hypoxia inducible factor-1alpha (HIF-1alpha) were used to compare retinas from the hypoxic and recovery groups to control animals that were not made hypoxic. SAM analysis was used to identify statistically significant changes in microarray data, and the bioinformatics programs GoMiner, Gene Set Enrichment Analysis (GSEA), and HiMAP were used to identify significant ontological categories and analyze the N-methyl-D-aspartate (NMDA) receptor interactome. HIF-1alpha protein, but not mRNA, was elevated up to 15-fold during hypoxia, beginning at 0.5 h, the shortest duration examined. Of the total of 1,178 genes examined by microarray, 119 were significantly upregulated following hypoxia. Of these, 86 were still significantly upregulated following recovery. However, 24 genes were significantly downregulated following hypoxia, with 12 still significantly downregulated after recovery. Of the 1035 genes that did not change with hypoxia, the expression of 36 genes was significantly changed after recovery. Ontological analyses showed significant upregulation of a large number of genes in the glutamate receptor family, including 3 of the 5 NMDA subunits. qRT-PCR analysis further corroborated these findings. Genes known to directly interact specifically with the NR1 subunit of the NMDA receptor were identified using HiMAP databases. GSEA analysis revealed that these genes were not affected by either hypoxia or altered after recovery. The identification of gene expression alterations as a function of hypoxia and recovery from hypoxia is important to understand the molecular mechanisms underlying retinal dysfunction associated with a variety of diseases. Gene changes were identified in hypoxic retina that could be linked to specific networks. Retinas recovering from hypoxia also showed distinct patterns of gene expression that were different from both normoxic control retinas and hypoxic retinas, indicating that hypoxia initiates a complex pattern of gene expression. Diseases of which hypoxia is a component may exhibit the several changes found here. Several potential therapeutic targets have been identified by our approach, including modulation of NMDA receptor expression and signaling, which until now have only been shown to play a role in responding to ischemia.


Coverage-dependent self-assembly of rubrene molecules on noble metal surfaces observed by scanning tunneling microscopy.
Coverage-dependent self-assembly of rubrene molecules on different noble metal surfaces, Au(111) and Au(100), Ag(111) and Ag(100), is presented. On Au(111), the homochiral supramolecular assemblies evolve with increasing rubrene coverage from very small structures composed of a few molecules, to honeycomb islets, and to one-dimensional chains of supramolecular pentamers. At higher coverage, the racemic mixture of molecules forms close-packed islands. On Au(100), chains of pentamers and two different types of densely packed islands are formed. On the Ag surfaces, exclusively close-packed islands are created, independently of the rubrene coverage. Moreover, the role of the chiral nature of the molecules in the self-assembly process is discussed, as well as the existence of different molecular conformers depending on the supramolecular assembled phase. The observed differences and similarities reflect the influence of the electronic properties and the geometric structure of the various substrates on molecular self-assembly.


Size/shape-controlled synthesis of colloidal CdSe quantum disks: ligand and temperature effects.
Size/shape-controlled colloidal CdSe quantum disks with zinc-blende (cubic) crystal structure were synthesized using air-stable and generic starting materials. The colloidal CdSe quantum disks were approximately square, and their lateral dimensions were varied between 20 and 100 nm with the thickness controlled between 1 and 3 nm, which resulted in sharp and blue-shifted UV-vis and PL peaks due to one-dimensional quantum confinement. The quantum disks were grown with either <001> or <111> direction, polar directions in the single crystalline disks, as the short axis, and both basal planes were terminated with Cd ions. These surface Cd ions were passivated with negatively charged fatty acid ligands to neutralize the net positive charges caused by the excess monolayer of Cd ions. The coordination of the Cd ions and carboxylate groups further enabled the close-packing monolayer of fatty acid ligands on each basal plane. The close packing of the hydrocarbon chains of fatty acids dictated the up temperature limit for synthesis of the colloidal quantum disks, and the low temperature limit was found to be related to the reactivity of the starting materials. Overall, a high Cd to Se precursor ratio, negative-charged fatty acid ligands with a long hydrocarbon chain, and a proper temperature range (approximately between 140 and 250 °C) were found to be needed for successful synthesis of the colloidal CdSe quantum disks.


Multiplex real-time PCR probe-based for identification of strains producing: OXA48, VIM, KPC and NDM.
The spread of multi-resistant enterobacteria, particularly carbapenem-resistant Enterobacteriaceae (CRE), in both community and hospital settings is a global problem. The phenotypic identification of CRE is complex, occasionally inconclusive and time consuming. However, commercially available molecular assays are very expensive, and many do not allow the simultaneous identification of all genetic markers of resistance that have been recognised in CRE (bla KPC, bla OXA-48, bla VIM and bla NDM). The aim of the present study is to describe a new test: a multiplex real time PCR probe-based assay designed for the simultaneous detection of KPC, OXA-48, VIM and NDM in a short time (no longer than 90 min from the extraction of DNA to detection). Our assay correctly identified 63 CRE isolates and all standard reference strains tested, in agreement with and extending the results of phenotypic identification tests; additionally, a KPC-VIM co-expressing Enterobacter aerogenes isolate was identified using the new assay, whereas traditional methods failed to detect it. The assay was also able to correctly detect 28 CRE-producers from 50 positive blood cultures, again detecting, in four specimens, the presence of CRE co-expressing KPC and VIM, which were only partially identified by traditional methods. Finally, when used directly on rectal swabs, the assay enabled the identification of CRE-carrier patients, for whom isolation is mandatory in a hospital setting.


Early Events in the Endoplasmic Reticulum Unfolded Protein Response.
The physiological consequences of the unfolded protein response (UPR) are mediated by changes in gene expression. Underlying them are rapid processes involving preexisting components. We review recent insights gained into the regulation of the endoplasmic reticulum (ER) Hsp70 chaperone BiP, whose incorporation into inactive oligomers and reversible AMPylation and de-AMPylation present a first line of response to fluctuating levels of unfolded proteins. BiP activity is tied to the regulation of the UPR transducers by a recently discovered cycle of ER-localized, J protein-mediated formation of a repressive IRE1-BiP complex, whose working we contrast to an alternative model for UPR regulation that relies on direct recognition of unfolded proteins. We conclude with a discussion of mechanisms that repress messenger RNA (mRNA) translation to limit the flux of newly synthesized proteins into the ER, a rapid adaptation that does not rely on new macromolecule biosynthesis.


Domestication shapes the pig gut microbiome and immune traits from the scale of lineage to population.
Animal ecology and evolution have long been known to shape host physiology, but more recently, the gut microbiome has been identified as a mediator between animal ecology and evolution and health. The gut microbiome has been shown to differ between wild and domestic animals, but the role of these differences for domestic animal evolution remains unknown. Gut microbiome responses to new animal genotypes and local environmental change during domestication may promote specific host phenotypes that are adaptive (or not) to the domestic environment. Because the gut microbiome supports host immune function, understanding the effects of animal ecology and evolution on the gut microbiome and immune phenotypes is critical. We investigated how domestication affects the gut microbiome and host immune state in multiple pig populations across five domestication contexts representing domestication status and current living conditions: free-ranging wild, captive wild, free-ranging domestic, captive domestic in research or industrial settings. We observed that domestication context explained much of the variation in gut microbiome composition, pathogen abundances and immune markers, yet the main differences in the repertoire of metabolic genes found in the gut microbiome were between the wild and domestic genetic lineages. We also documented population-level effects within domestication contexts, demonstrating that fine scale environmental variation also shaped host and microbe features. Our findings highlight that understanding which gut microbiome and immune traits respond to host genetic lineage and/or scales of local ecology could inform targeted interventions that manipulate the gut microbiome to achieve beneficial health outcomes.


Processing changes across reading encounters.
Five experiments examined changes in the processing of a text across reading encounters. Experiment 1 showed that reading speed increased systematically across encounters, with no loss in the extensiveness of analyses of the printed text, as indicated by the ability to detect nonword errors embedded within that passage. Experiment 2 replicated this improved reading fluency with experience and showed that it occurred even with typescript changes across trials, thus indicating that a primed visual operations explanation cannot account for the effect. The third and fourth experiments then extended the study of the familiarity effect to higher level processing, as indicated by the detection of word errors. Familiarity facilitated the detection of these violations at the syntactic-semantic levels. Finally, Experiment 5 showed that these higher level violations continued to be well detected over a series of reading encounters with the same text. The results indicate that prior experience improves reading speed, with no attenuation of analysis of the printed words or of the passage's message.


Enhancing performance of berseem clover genotypes with better harvesting management through farmers' participatory research at smallholder farms in Punjab.
A field study was conducted on smallholder farmer fields between 2012 to 2014 to evaluate the performance of cv. Agaitti Berseem-2002, against local landraces exchanged between farmers (LBF1) or available from local markets (LBM1). The effects of genotype and harvesting regimen on forage production, quality and seed production were evaluated. Significant differences (P < 0.05) among genotypes and cutting treatments were recorded for forage and seed yields, and forage quality across all research sites in both years. Maximum cumulative fresh forage (89.7 t/ha) and dry matter (DM; 13.4 t/ha) yields were obtained with Agaitti Berseem-2002 when harvesting occurred five times over the season. However, maximum seed yield (1048 kg/ha) with higher 1000-seed weight (3.63 g) were obtained if forage was only harvested three times and the crop then left for seed set. Agaitti Berseem-2002 also produced forage with the higher crude protein content (27%), DM digestibility (69%), digestible organic matter (DM basis; 65%) and metabolizable energy content (10%) compared to the local landraces (LBF1 and LBM1). Therefore, the harvesting regimen for greatest economic return which produced optimum fresh and DM forage yields of highest nutritive values and maximum seed yield, were comprised of taking three forage cuts (at 65, 110 and 150 days after sowing) prior to seed harvest.


UGT1A1 *6 polymorphism predicts outcome in elderly patients with relapsed or refractory diffuse large B-cell lymphoma treated with carboplatin, dexamethasone, etoposide and irinotecan.
The uridine diphosphate glucuronosyltransferase (UGT) gene 1A1*6 polymorphism, which affects irinotecan metabolism, has been associated with improved survival in lymphoma patients treated with of carboplatin, dexamethasone, etoposide and irinotecan (CDE-11). This study assessed the efficacy of CDE-11 relative to the UGT1A1*6 polymorphism in 27 elderly patients with relapsed or refractory diffuse large B-cell lymphoma who were ineligible for high-dose chemotherapy plus autologous stem cell transplantation. The 2-year survival rate after initial CDE-11 treatment was significantly higher in patients with than without UGT1A1*6 (57% vs. 5%). The most common grade 4 adverse event in patients with the UGT1A1*6 genotypes was neutropenia (88.9%), but there were no gastrointestinal adverse events or treatment-related deaths. Disease progression was the most frequent cause of death. CDE-11 was well tolerated and provided clinical benefit to elderly patients with relapsed or refractory diffuse large B-cell lymphoma. The response to CDE-11 likely correlated with UGT1A1*6 polymorphisms, but further prospective studies are warranted to optimize irinotecan-based chemotherapies relative to UGT1A1 polymorphism.


Bidirectional electron transfer in photosystem I: determination of two distances between P700+ and A1- in spin-correlated radical pairs.
The spin-correlated radical pair [P(700)(+)A(1)(-)] gives rise to a characteristic "out-of-phase" electron spin-echo signal. The electron spin-echo envelope modulation (ESEEM) of these signals has been studied in thylakoids prepared from the wild-type strain of Chlamydomonas reinhardtii and in two site-directed mutants, in which the methionine residue which acts as the axial ligand to the chlorin electron acceptor A(0) has been substituted with a histidine either on the PsaA (PsaA-M684H) or the PsaB (PsaB-M664H) reaction center subunits. The analysis of the time domain ESEEM provides information about the spin-spin interaction in the [P(700)(+)A(1)(-)] radical pair, and the values of the dipolar (D) and the exchange (J) interaction can be extracted. From the distance dependence of the dipolar coupling term, the distance between the unpaired electron spin density clouds of the primary donor P(700)(+) and the phyllosemiquinone A(1)(-) can be determined. The [P(700)(+)A(1)(-)] ESEEM spectrum obtained in wild-type thylakoids can be reconstructed using a linear combination of the spectra measured in the PsaA and PsaB A(0) mutants, demonstrating that electron transfer resulting in charge separation is occurring on both the PsaA and PsaB branches. The [P(700)(+)A(1B)(-)] distance in the point dipole approximation in the PsaA-M684H mutant is 24.27 +/- 0.02 A, and the [P(700)(+)A(1A)(-)] distance in the PsaB-M664H mutant is 25.43 +/- 0.01 A. An intermediate value of 25.01 +/- 0.02 A is obtained in the wild-type membranes which exhibit both spin-polarized pairs.


Importance of temporal flow gradients and integrin alphaIIbbeta3 mechanotransduction for shear activation of platelets.
Disturbances of blood flow play an important role in promoting platelet activation and arterial thrombus formation in stenosed, injured, atherosclerotic arteries. To date, glycoprotein Ib (GPIb) has been considered the primary platelet mechanosensory receptor, responding to increased shear with enhanced adhesive and signaling function. We demonstrate here that von Willebrand factor-GPIb interaction is inefficient at inducing platelet activation even when platelets are exposed to very high wall shear stresses (60 dyn/cm(2)). Rapid platelet activation under flow was only observed under experimental conditions in which transiently adherent platelets were exposed to sudden accelerations in blood flow. Platelet responsiveness to temporal shear gradients was integrin alpha(IIb)beta(3)-dependent and occurred only on a von Willebrand factor substrate, as platelets forming integrin alpha(IIb)beta(3) adhesive contacts with immobilized fibrinogen were unresponsive to sudden increases in shear. The calcium response induced by temporal shear gradients was distinct from previously identified integrin alpha(IIb)beta(3) calcium responses in terms of its transient nature, its requirement for platelet co-stimulation by the P2Y(1) purinergic ADP receptor, and its dependence on the influx of extracellular calcium. Our studies demonstrate a key role for temporal shear gradients in promoting platelet activation. Moreover, they define for the first time the involvement of P2Y receptors in integrin mechanotransduction.


Lasting power of new clotting proteins.
Hemophilia is a genetic disease caused by a deficiency of one of the coagulation proteins. The term usually refers to either hemophilia A, factor VIII (FVIII), with an incidence of ∼1 in 5000 male births, or hemophilia B, factor IX (FIX), with an incidence of ∼1 in 30 000 male births. When severe, the disease leads to spontaneous life-threatening bleeding episodes. Current therapy requires frequent intravenous infusions of therapeutic factor concentrates. Most patients administer the infusions at home every few days and must limit their physical activities to avoid bleeding when the factor activity levels are below normal. In March 2014, a new therapeutic FIX preparation was approved for clinical use in Canada and the United States and, in June 2014, a new FVIII preparation was approved for clinical use in the United States. Over the next couple of years, other new factor products for FIX, FVIIa, and FVIII, which are currently in late stages of clinical trials, will likely also be approved. These new factors have been engineered to extend their half-life in circulation, thus providing major therapeutic advances for patients with hemophilia primarily by allowing treatment with fewer infusions per month. In the clinical trials so far, >500 patients have successfully used these extended half-life products regularly for >1 year to prevent spontaneous bleeding, to treat successfully any bleeding episodes, and to provide effective coagulation for major surgery. Essentially all infusions were well tolerated and effective. These promising new therapies should allow patients to use fewer infusions to maintain appropriate clotting factor activity levels in all clinical settings.


A facile method for the synthesis of copper-cysteamine nanoparticles and study of ROS production for cancer treatment.
Copper-cysteamine (Cu-Cy) is a novel sensitizer that can be excited by ultraviolet (UV) light, microwave (MW), ultrasound, and X-rays to generate highly toxic reactive oxygen species (ROS) for cancer cell destruction. The purpose of this study is to present a facile method for the synthesis of Cu-Cy nanoparticles. Interestingly, we were able to decrease both the stirring and heating time by about 24 and 6 times, respectively, thus making Cu-Cy nanoparticles more economical than what was reported before. 1,4-Diazabicylo[2.2.2]octane (DABCO), a well-known singlet oxygen quencher, showed that the majority of ROS produced by Cu-Cy nanoparticles upon UV and MW exposure were singlet oxygen. Moreover, ROS generated by Cu-Cy nanoparticles upon UV and MW exposure were confirmed by a known ROS tracking agent, dihydrorhodamine 123, further serving as an additional piece of evidence that Cu-Cy is a promising ROS generating agent to destroy cancer cells as well as bacteria or viruses by a radical therapeutic approach. Additionally, for the first time, the hydroxyl radical (˙OH) produced by Cu-Cy nanoparticles upon MW activation was proved by a photoluminescence (PL) technique using coumarin as a probe molecule. Remarkably, newly synthesized nanoparticles were found to be much more effective for producing ROS and killing cancer cells, suggesting that the new method may have increased the reactivity of the Cu-Cy nanoparticles due to an overall size reduction. Overall, the new method not only reduced the synthesis time but also enhanced the effectiveness of Cu-Cy nanoparticles for photodynamic therapy.


Prevalence, distribution, and host range of Peste des petits ruminants virus, Turkey.
Peste des petits ruminants virus (PPRV, genus Morbillivirus), which causes a severe disease in sheep and goats, has only recently been officially declared to be present in Turkey. We carried out a study to determine the prevalence, distribution, and host range of PPRV in Turkey. A total of 1,607 animals, reared in 18 different locations, were monitored for the presence of antibodies to PPRV and the related virus of large ruminants, Rinderpest virus (RPV). Only two farms had animals that were free of antibody responses to either disease. Prevalence for PPRV infection varied (range 0.87%-82.6%) and was higher in sheep (29.2%) than in goats (20%). The overall antibody responses to PPRV and RPV were 22.4% and 6.28%, respectively. Two PPRVs of lineage 4, which comprises many other PPRVs whose origins are in the Middle East, the Arabian Peninsula, and southern Asia, were isolated from Turkish sheep.


miRNA-96 suppresses KRAS and functions as a tumor suppressor gene in pancreatic cancer.
Therapeutic applications of microRNA (miRNA) in KRAS-driven pancreatic cancers might be valuable, but few studies have explored this area. Here, we report that miR-96 directly targets the KRAS oncogene and functions as a tumor-suppressing miRNA in pancreatic cancer cells. Ectopic expression of miR-96 through a synthetic miRNA precursor inhibited KRAS, dampened Akt signaling, and triggered apoptosis in cells. In human clinical specimens, miR-96 was downregulated or deleted where an association with KRAS elevations was observed. In vitro and in vivo assays established that miR-96 decreased cancer cell invasion and migration and slowed tumor growth in a manner associated with KRAS downregulation. Our findings identify miR-96 as a potent regulator of KRAS, which may provide a novel therapeutic strategy for treatment of pancreatic cancer and other KRAS-driven cancers.


Longitudinal analysis of maternal serum leptin levels during pregnancy, at birth and up to six weeks after birth: relation to body mass index, skinfolds, sex steroids and umbilical cord blood leptin levels.
Leptin is an important regulator of body fat mass and energy expenditure during adult life. The mechanisms by which maternal and fetal weight are regulated during pregnancy are poorly understood. In order to gain more insight into a potential role of leptin during gestation, a prospective, longitudinal study was carried out to measure leptin concentrations in maternal serum of 29 healthy women during pregnancy up to 6 weeks after birth and also in umbilical cord blood of their newborns. Leptin concentrations were measured using a specific RIA. In addition, estradiol, testosterone, and sex hormone binding globulin were determined using commercially available RIAs. The mothers' skinfolds were determined at four sites using a Holtain caliper. Leptin levels increased continuously during pregnancy and reached 25.8 +/- 14.7 ng/ml at 38-40 weeks. At birth, leptin concentrations were 23.5 +/- 15.4 ng/ml. Three days after delivery a significant decrease of leptin levels to 10.6 +/- 6.0 ng/ml was observed. Six weeks after birth the leptin concentration in maternal serum was 13.8 +/- 8.6 ng/ml. At birth, maternal serum levels were significantly higher than levels in cord blood and did not correlate with leptin levels in cord blood or neonatal weight. Furthermore, leptin levels did not correlate with maternal sex steroids and sex hormone binding globulin levels. At 6-8 weeks of pregnancy, maternal leptin serum levels correlated significantly with BMI (r = 0.81). The correlation coefficients (leptin vs. BMI) dropped with increasing gestational age and at birth only a poor correlation persisted (r = 0.50). Six weeks after birth there was again a high correlation between leptin levels in maternal serum and BMI (r = 0.76). Subscapular skinfold thickness was correlated to leptin concentrations in maternal serum during the whole period of the investigation. In conclusion, maternal leptin levels continuously increased from 6-8 weeks up to 38-40 weeks of pregnancy. Maternal leptin levels decreased dramatically after birth. Six weeks after delivery, leptin levels were comparable to the values measured at the beginning of pregnancy. We hypothesize that leptin might play an important role during pregnancy and fetal development.


Splinkerette PCR for mapping transposable elements in Drosophila.
Transposable elements (such as the P-element and piggyBac) have been used to introduce thousands of transgenic constructs into the Drosophila genome. These transgenic constructs serve many roles, from assaying gene/cell function, to controlling chromosome arm rearrangement. Knowing the precise genomic insertion site for the transposable element is often desired. This enables identification of genomic enhancer regions trapped by an enhancer trap, identification of the gene mutated by a transposon insertion, or simplifying recombination experiments. The most commonly used transgene mapping method is inverse PCR (iPCR). Although usually effective, limitations with iPCR hinder its ability to isolate flanking genomic DNA in complex genomic loci, such as those that contain natural transposons. Here we report the adaptation of the splinkerette PCR (spPCR) method for the isolation of flanking genomic DNA of any P-element or piggyBac. We report a simple and detailed protocol for spPCR. We use spPCR to 1) map a GAL4 enhancer trap located inside a natural transposon, pinpointing a master regulatory region for olfactory neuron expression in the brain; and 2) map all commonly used centromeric FRT insertion sites. The ease, efficiency, and efficacy of spPCR could make it a favored choice for the mapping of transposable element in Drosophila.