Targeted Delivery of Stk25 Antisense Oligonucleotides to Hepatocytes Protects Mice Against Nonalcoholic Fatty Liver Disease.
Nonalcoholic fatty liver disease (NAFLD) and nonalcoholic steatohepatitis (NASH) are emerging as leading causes of liver disease worldwide. Currently, no specific pharmacologic therapy is available for NAFLD/NASH, which has been recognized as one of the major unmet medical needs of the 21st century. Our recent studies in genetic mouse models, human cell lines, and well-characterized patient cohorts have identified serine/threonine protein kinase (STK)25 as a critical regulator of hepatic lipid partitioning and NAFLD/NASH. Here, we studied the metabolic benefit of liver-specific STK25 inhibitors on NAFLD development and progression in a mouse model of diet-induced obesity. We developed a hepatocyte-specific triantennary N-acetylgalactosamine (GalNAc)-conjugated antisense oligonucleotide (ASO) targeting Stk25 and evaluated its effect on NAFLD features in mice after chronic exposure to dietary lipids. We found that systemic administration of hepatocyte-targeting GalNAc-Stk25 ASO in obese mice effectively ameliorated steatosis, inflammatory infiltration, hepatic stellate cell activation, nutritional fibrosis, and hepatocellular damage in the liver compared with mice treated with GalNAc-conjugated nontargeting ASO, without any systemic toxicity or local tolerability concerns. We also observed protection against high-fat-diet-induced hepatic oxidative stress and improved mitochondrial function with Stk25 ASO treatment in mice. Moreover, GalNAc-Stk25 ASO suppressed lipogenic gene expression and acetyl-CoA carboxylase protein abundance in the liver, providing insight into the molecular mechanisms underlying repression of hepatic steatosis. This study provides in vivo nonclinical proof-of-principle for the metabolic benefit of liver-specific inhibition of STK25 in the context of obesity and warrants future investigations to address the therapeutic potential of GalNAc-Stk25 ASO in the prevention and treatment of NAFLD.


Single-stranded DNA binding protein from bacteriophage cf: characterization, gene localization and protein-ssDNA complex.
The single-stranded DNA binding protein from the filamentous bacteriophage cf has been purified and characterized. The first 12 amino acids, resulting from the N-terminal amino acid sequencing analysis of the protein, agree with an open reading frame (ORF) on the cf genome. The ORF contains 294 bp and codes for a 98 a.a. protein of molecular weight 10.8 kDa, consistent with the result from the denaturing protein gel analysis. The protein appears to be a homodimer as evident from the apparent molecular weight of about 22 kDa obtained from native protein gel analysis. The gene location of the protein has been identified as gene V of the cf single stranded genome, same as that from the M13 phage. The GVP of cf shows a strong sequence homology to the ssDNA binding proteins of Ff, IKe and Pf3 filamentous phages. The DNA binding wing of GVP, conserved among the filamentous phages, has been predicted for cf. To further characterize the protein, the GVP-ssDNA complex of cf has been purified from the infected host (Xanthomonas campestris pv. citri) by density gradient centrifugation. Transmission electron microscopy (TEM) images of the complex showed that it is about 1200 nm in length and 9 nm in diameter and it has a highly regular morphology with a central groove shadow running along the entire structure, but without any apparent helical pattern seen in the M13 complex. The GVP-ssDNA complex of cf seems more rigid than that of M13. Our computer modeling study suggested that this difference between the two complexes may be due to the additional 11 or 12 amino acids at the C-terminal end of the cf-GVP.


Bridging the Homogeneous-Heterogeneous Divide: Modeling Spin for Reactivity in Single Atom Catalysis.
Single atom catalysts (SACs) are emergent catalytic materials that have the promise of merging the scalability of heterogeneous catalysts with the high activity and atom economy of homogeneous catalysts. Computational, first-principles modeling can provide essential insight into SAC mechanism and active site configuration, where the sub-nm-scale environment can challenge even the highest-resolution experimental spectroscopic techniques. Nevertheless, the very properties that make SACs attractive in catalysis, such as localized d electrons of the isolated transition metal center, make them challenging to study with conventional computational modeling using density functional theory (DFT). For example, Fe/N-doped graphitic SACs have exhibited spin-state dependent reactivity that remains poorly understood. However, spin-state ordering in DFT is very sensitive to the nature of the functional approximation chosen. In this work, we develop accurate benchmarks from correlated wavefunction theory (WFT) for relevant octahedral complexes. We use those benchmarks to evaluate optimal DFT functional choice for predicting spin state ordering in small octahedral complexes as well as models of pyridinic and pyrrolic nitrogen environments expected in larger SACs. Using these guidelines, we determine Fe/N-doped graphene SAC model properties and reactivity as well as their sensitivities to DFT functional choice. Finally, we conclude with broad recommendations for computational modeling of open-shell transition metal single-atom catalysts.


Functional characterization of novel telomerase RNA (TERC) mutations in patients with diverse clinical and pathological presentations.
Functional characterization of heterozygous TERC (telomerase RNA component) and TERT (telomerase reverse transcriptase) mutations found in autosomal dominant dyskeratosis congenita (DC) and aplastic anemia (AA) shows that telomerase function is defective and that this is associated with short telomeres. This leads to reduced cell longevity with maximal impact on tissues with high proliferate potential. The aim of this study was to establish the role of TERC in the pathophysiology of uncharacterized patients with AA with some features of DC. The TERC gene was screened for mutations by denaturing high performance liquid chromatography. To determine the functional significance of TERC mutations telomerase activity was assessed in an in vitro (TRAP) assay and telomere length of patients' samples was determined using Southern blot analysis. RESULTS This study led to the identification of four novel TERC mutations (G178A, C180T, D52-86 and G2C) and a recurrent TERC mutation (D110-113GACT). Two of the de novo TERC mutations (G178A and C180T) found uniquely produce a clinical phenotype in the first generation, differing from previously published cases in which individuals in the first generation are usually asymptomatic. Curiously these mutations are located near the triple-helix domain of TERC. We also observed that the recurrent D110-113GACT can present with AA, myelodysplasia or leukemia. The D52-86 is associated with varied phenotypes including pulmonary disease (pulmonary fibrosis) as the first presentation. In summary, this study reports the functional characterization of several novel TERC mutations associated with varied hematologic and extra-hematologic presentations.


Axillobifemoral bypass: a brief surgical and historical review.
Peripheral artery disease (PAD) occurs when plaque accumulates in the arterial system and obstructs blood flow. Narrowing of the abdominal aorta and the common iliac arteries due to atherosclerotic plaques restricts blood supply to the lower limbs. Clinically, the lower limb symptoms of PAD are intermittent claudication, discoloration of the toes, and skin ulcers, all due to arterial insufficiency. Surgical revascularization is the primary mode of treatment for patients with severe limb ischemia. The objective of the surgical procedure is to bypass a blockage in an occluded major vessel by constructing an alternate route for blood flow using an artificial graft. This article presents information on aortoiliac reconstruction, with an emphasis on axillobifemoral bypass grafting.


[The role of the adreno-cholinergic interaction in the pulmonary hemodynamics changes following myocardial ischemia].
In acute experiments in anesthetized rabbits the pulmonary hemodynamics changes were studied following 60 s myocardial ischemia in the region of the descendent left coronary artery in control state and after the blockade of M- or N-cholinoreceptors and acetylcholine infusion. Following myocardial ischemia in control animals the pulmonary artery pressure and flow decreased, the pulmonary vascular resistance was not changed. Following myocardial ischemia after the blockade of M-cholinoreceptors by atropine the changes of pulmonary hemodynamics were the same as in control animals, the cardiac output decreased twice as more as in control animals. Following myocardial ischemia after the blockade of N-cholinoreceptors by hexamethonium the pulmonary hemodynamics changes were the same as in the control rabbits. Following myocardial ischemia after the acetylcholine infusion the pulmonary artery flow decreased more than the cardiac output, the pulmonary vascular resistance was diminished. The disbalance of the cardiac output and pulmonary artery flow changes has revealed the significance of the adreno-cholinergic interaction in the changes of the pulmonary vessels capacitance and resistive functions following myocardial ischemia.


Structure, chromosome mapping, and expression of the mouse Lyl-1 gene.
The mouse Lyl-1 gene was cloned and shown to consist of four exons with extensive nucleotide and structural homology to the human LYL1 gene. The Lyl-1 gene was localized to the central region of mouse chromosome 8 which defines a new region of synteny with human chromosome 19p. The predicted mouse Lyl-1 protein is 78% identical to human LYL1. The region of highest similarity occurs in the basic DNA binding and helix-loop-helix dimerization motifs which are nearly identical in mouse and man differing by only one conservative amino acid substitution. Expression of the Lyl-1 gene was found to be low in murine spleen and undetectable in other tissues by Northern blot analysis. In lymphoid cell lines, Lyl-1 was expressed in most B lineage cells but downregulated during terminal differentiation and was not expressed in most T lineage cells. In a human T ALL cell line carrying a translocation that juxtaposed LYL1 with the beta TCR gene, the translocated LYL1 gene was transcriptionally active whereas the nontranslocated gene was transcriptionally silent. We conclude that LYL1 has the properties of a lineage- and differentiation-specific HLH protein that contributes to T-cell neoplasia through its deregulated expression following chromosomal translocation.


Classification of Continuous ECG Segments - Performance Analysis of a Deep Learning Model.
Classification of electrocardiogram (ECG) signals plays an important role in the diagnosis of heart diseases. It is a complex and non-linear signal, which is the first option to preliminary identify specific pathologies/conditions (e.g., arrhythmias). Currently, the scientific community has proposed a multitude of intelligent systems to automatically process the ECG signal, through deep learning techniques, as well as machine learning, where this present high performance, showing state-of-the-art results. However, most of these models are designed to analyze the ECG signal individually, i.e., segment by segment. The scientific community states that to diagnose a pathology in the ECG signal, it is not enough to analyze a signal segment corresponding to the cardiac cycle, but rather an analysis of successive segments of cardiac cycles, to identify a pathological pattern.In this paper, an intelligent method based on a Convolutional Neural Network 1D paired with a Multilayer Perceptron (CNN 1D+MLP) was evaluated to automatically diagnose a set of pathological conditions, from the analysis of the individual segment of the cardiac cycle. In particular, we intend to study the robustness of the referred method in the analysis of several simultaneous ECG signal segments. Two ECG signal databases were selected, namely: MIT-BIH Arrhythmia Database (D1) and European ST-T Database (D2). The data was processed to create datasets with two, three and five segments in a row, to train and test the performance of the method. The method was evaluated in terms of classification metrics, such as: precision, recall, f1-score, and accuracy, as well as through the calculation of confusion matrices.Overall, the method demonstrated high robustness in the analysis of successive ECG signal segments, which we can conclude that it has the potential to detect anomalous patterns in the ECG signal. In the future, we will use this method to analyze the ECG signal coming in real-time, acquired by a wearable device, through a cloud system.Clinical Relevance-This study evaluates the potential of a deep learning method to classify one or several segments of the cardiac cycle and diagnose pathologies in ECG signals.


Low Dose Brain Irradiation Reduces Amyloid-β and Tau in 3xTg-AD Mice.
We have previously reported that low doses of external beam ionizing irradiation reduced amyloid-β (Aβ) plaques and improved cognition in APP/PS1 mice. In this study we investigated the effects of radiation in an age-matched series of 3xTg-AD mice. Mice were hemibrain-irradiated with 5 fractions of 2 Gy and sacrificed 8 weeks after the end of treatment. Aβ and tau were assessed using immunohistochemistry and quantified using image analysis with Definiens Tissue Studio. We observed a significant reduction in Aβ plaque burden and tau staining; these two parameters were significantly correlated. This preliminary data is further support that low doses of radiation may be beneficial in Alzheimer's disease.


A Primordial Attachment to the Nation? French and Irish Workers and Trade Unions in Past EU Referendum Debates.
We aim to contribute to a better understanding of the dynamics that are driving EU politicization and the rising Euroscepticism of workers and unions in the public sphere. One explanation frames the rise in Euroscepticism in cultural terms, emphasizing workers' alleged primordial attachment to their nation. A second uses socioeconomic frames, linking growing Euroscepticism to the increasingly neo-liberal direction of the EU. The weight of these competing frames in the referendum campaigns on the EU Constitution in France and the Lisbon Treaty and the Fiscal Treaty in Ireland cannot be measured easily, as the categorization of a phrase as socioeconomic or cultural is in itself subject to political classification struggles. We therefore presents the findings of an inductive lexical analysis of all Irish Times, all Le Monde and all worker- or union-related articles published in almost all national media outlets during the mentioned referendum debates. This was made possible by the Alceste software package that allowed us to analyse very large corpuses of articles inductively. Our analysis reveals that socioeconomic terms dominated policy debates in both countries. The findings question existing EU politicization studies that were measuring the salience of different frame types by deductive analysis.


[Status and developmental prospects of the medical proton channel in the synchrocyclotron in Gatchina].
The medical proton beam on the synchrocyclotron in Gatchina modernized in 1980, is characterized by a simplicity of the beam adjustment that amounts to standard operations, the time of adjustment reduced considerably. The optical scheme ensures a good reproducibility of all the beam parameters from session to session. A large "zone of convergence" makes it possible to select a necessary irradiation field size with the diameter from 3 to 10 mm. A new medical complex with a 2-room premises for irradiation is envisaged on the basis of the "second beam" to expand the medical program. The piercing technique of moving field irradiation with protons at an energy of 1 Gy and the utilization of the convergent beam ensure the ratio of the focal dose to the surface dose 200:1.


Intrasubunit signal transduction by the aspartate chemoreceptor.
Receptors that transmit signals across cell membranes are typically composed of multiple subunits. To test whether subunit interactions are required for transmembrane signaling by the bacterial aspartate receptor, dimers were constructed with (i) two full-length subunits, (ii) one full-length subunit and one subunit lacking the cytoplasmic domain, or (iii) one full-length subunit and one subunit lacking both the cytoplasmic and the transmembrane domains. Methylation of the cytoplasmic domain of all three receptor constructs was stimulated by the binding of aspartate. These findings demonstrate that transmembrane signaling does not require interactions between cytoplasmic or transmembrane domains of adjacent subunits and suggest that signaling occurs via conformational changes transduced through a single subunit.


[Effects on serum glucose and lipids in chronic exposure to lanthanum chloride in rat].
To explore the effects of serum glucose and lipids by on chronically lanthanum exposure in rat. The Wistar rats were treated with oral exposure dose 0.1, 2 and 40 mg/kg of lanthanum chloride (LaCl(3)) respectively, after 90 days the rats were sacrificed and the blood was collected for measuring the glycosylated hemoglobin A (HbA1c), the serum was used for measuring glucose, total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C) levels. There were no any differences among the control and 3 dose LaCl(3) exposed rats on the blood HbA1c and serum Glu, TG and LDL-C levels (P > 0.05). The serum TC in 0.1 and 2 mg/kg LaCl(3) dose group rats were (1.38 +/- 0.14) mmol/L and (1.37 +/- 0.26) mmol/L respectively. It was lower than that of the controls (1.57 +/- 0.14) mmol/L significantly (P < 0.05), the serum HDL-C in 0.1 mg/kg dose group rats was (0.79 +/- 0.12) mmol/L and obviously lower than that of control group rats (0.93 +/- 0.10) mmol/L (P < 0.05). 0.1 - 40 mg/kg LaCl(3) chronically exposed have not greater effect on serum glucose, TG and LDL-C levels in rats, but the lower dose LaCl(3) chronic exposure might cause serum TC and HLD-C level decreasing.


Generation of amplitude-shift-keying optical signals using silicon microring resonators.
We demonstrate the generation of amplitude-shift-keying (ASK) optical signals using a system of parallel microring resonators. By independently modulating two symmetric microring resonators arranged in a Mach-Zehnder configuration, we realize the generation of three levels. The proposed scheme can be extended to any number of logic levels, which effectively increases the data rate of an optical link using slower modulators. Here, we separately utilize thermo-optic and ultrafast all-optical modulation schemes to generate ASK signals on a silicon photonic chip.


Economic Advantages of Telehealth and Virtual Health Practitioners: Return on Investment Analysis.
Telehealth is a disruptive modality that challenges the traditional model of having a clinician or patient physically present for an appointment. The benefit is that it offers the opportunity to redesign the way services are offered. For instance, a virtual health practitioner can provide videoconference consultations while being located anywhere in the world that has internet. A virtual health practitioner also obviates the issues of attracting a specialist medical workforce to rural areas, and allows the rural health service to control the specialist services that they offer. The aim of this research was to evaluate the economic effects of 3 different models of care on rural and metropolitan hospital sites. The models of care examined were patient travel, telehealth using videoconferencing, and employment of a virtual health practitioner by a rural site. Using retrospective activity data for 3 years, a return on investment (ROI) analysis was undertaken from the perspective of a rural site and metropolitan partner site using a telehealth orthopedic fracture clinic as an example. Further analysis was conducted to calculate the number of patients that would be required to attend the clinic in each model of care for the sites to break even. The only service model that resulted in a positive ROI for the rural site over the 3-year period was the virtual health practitioner model. The breakeven analysis demonstrated that the rural site required the lowest number of patients to recoup costs in the virtual health practitioner model of care. The rural site was unable to recoup its costs within the travel model due to the lack of opportunity for reimbursement for services and the requirement to cover the cost of travel for patients. Our model demonstrated that rural health care providers can increase their ROI by employing a virtual health practitioner.


Effect of hexythiazox and spiromesifen resistance on the life cycle of the predatory mite Neoseiulus californicus (Acari: Phytoseiidae).
Neoseiulus californicus (McGregor) (Acari: Phytoseiidae) is one of the most important and effective predators of Tetranychus spp. (Acari: Tetranychidae). This study analyzed the effects of hexythiazox and spiromesifen resistance on biological characteristics of N. californicus. Pre-oviposition, oviposition, post-oviposition times, adult lifespan, total number of eggs laid per female, net reproductive rate (R0), intrinsic rate of increase (rm), doubling time (DT), mean generation time (T) and finite rate of increase (λ) were compared among three populations: 63.9-fold hexythiazox resistant (HEX14), 53.6-fold spiromesifen resistant (SPR13) and a susceptible base population. Pre-oviposition and oviposition times, mean number of eggs/female, adult lifespan, R0 and rm values were all significantly higher for the two resistant populations than for the susceptible population. Life tables of HEX14, SPR13 and the susceptible population showed that R0 was 35.0, 26.5 and 19.4 (females/female generation), rm was 0.35, 0.32 and 0.30 (females/female/day), DT was 1.92, 2.07 and 2.27 days, T was 9.8, 9.8 and 9.7 days, and λ was 1.43, 1.39 and 1.35 (individuals/female/day), respectively.


The Impact of Antiepileptic Drugs on Vitamin Levels in Epileptic Patients.
The impact of antiepileptics on serum vitamin levels is controversial and uncertain. With no clear conclusions on the impact of antiepileptics on serum levels of vitamins, there is a need for further clinical studies in order to ascertain the impact of old and newer antiepileptic drugs on serum levels of vitamins in epileptic patients, thus accomplishing a suitable usage of vitamins supplementation. The intention of the present research is to confirm the hypothesis of whether or not vitamin levels are altered with antiepileptic drugs. The study also aims to reveal which vitamin levels are particularly more altered, are vitamin levels affected by gender and the type and number of antiepileptics used. The present research was piloted in collaboration with the Department of Neurology in Qilu Hospital of Shandong University. A total of 63 serum samples of epileptic patients receiving antiepileptics as monotherapy or polytherapy were requested for analysis of nine vitamin serum levels. Total nine vitamins (B1, B2, B6, B9, B12, A, C, D and E) in epileptic patients receiving antiepileptic drugs were analyzed. The serum results of all vitamins were compiled and evaluated with SPSS. It was alarmingly found that serum levels of vitamin D were particularly very low in almost all (90%) epileptic patients in this study. Notably, serum levels of vitamin C and vitamin B1 were also below reference range in 72% and 46% epileptic patients, respectively. The remaining vitamins were almost in reference range for most of the patients. In our study, mean and frequency of vitamin D, C and B1 levels do not vary too much among different gender groups. The patients receiving newer antiepileptic drugs displayed a slightly increased serum vitamin D levels in comparison to the patients receiving older antiepileptic drugs. We found low vitamin D, C and B1 serum levels in patients who were on monotherapy as in comparison with patients on polytherapy. The most significant and surprising finding of this study revealed that serum vitamin D levels in particular were very low in almost all patients and in some patients' vitamin B1 serum levels were also below the reference range. More importantly, it is first time reported here that vitamin C serum levels were also below reference range in the majority of these Chinese epileptic patients. It is recommended that all these vitamins should be regularly monitored in addition to therapeutic drug monitoring of antiepileptic drugs. Additional clinical trials are required for further evaluation. It is also recommended that epileptic patients with low serum levels of these vitamins may be prescribed vitamins supplementations with antiepileptic drugs in order to control their seizures more effectively and efficiently.


Central adiposity and associated lifestyle factors in Cree children.
Aboriginal children are prone to central adiposity (CA), a component of the metabolic syndrome. The objective of this study was to determine if lifestyle factors were associated with CA in Canadian Cree children. Children aged 9-12 years were classified as having CA if their waist circumference met or exceeded the 85th percentile of the NHANES III reference. Weight status was determined using the CDC growth reference, dietary intake using three 24 h dietary recalls, physical activity using pedometers, and fitness by completion of the 20 m shuttle run test. Of the 178 children (79% participation rate), 32.6% were normal weight, 23.6% were overweight, and 43.8% were obese. Half (52.2%) of the children had CA (97.4% of obese children, 35.7% of overweight children, and 2.2% of normal weight children). Waist circumference was negatively correlated with pedometer step counts (r = -0.187, p = 0.012) and shuttle run time (r = -0.508, p < 0.001). In children with CA, waist circumference was positively correlated with sweetened beverage intake (r = 0.250, p = 0.016). The odds ratio (adjusted for age and sex) for CA for children consuming 3 or more fruits and vegetables per day was 0.43 (95% CI 0.18 - 0.98), for meeting step recommendations for a healthy body weight was 0.45 (95% CI 0.24 - 0.84), and for relative fitness was 0.12 (95% CI 0.04 - 0.33). CA was prevalent in children who were overweight and obese. Preventive strategies might include promoting a healthy diet, physical activity, and fitness.


MicroRNA miR-252 targets mbt to control the developmental growth of Drosophila.
Developmental growth is an intricate process involving the coordinated regulation of the expression of various genes, and microRNAs (miRNAs) play crucial roles in diverse processes throughout animal development. The ecdysone-responsive miRNA, miR-252, is normally upregulated during the pupal and adult stages of Drosophila development. Here, we found that overexpression of miR-252 in the larval fat body decreased total tissue mass through a reduction in both cell size and cell number, causing a concomitant decrease in larval size. Furthermore, miR-252 overexpression led to a delayed larval-to-pupal transition with defective anterior spiracle eversion, as well as a decrease in adult size and mass. Conversely, adult flies lacking miR-252 showed an increase in mass compared with control flies. We found that miR-252 directly targeted mbt, encoding a p21-activated kinase, to repress its expression. Notably, co-overexpression of mbt rescued the developmental and growth defects associated with miR-252 overexpression, indicating that mbt is a biologically relevant target of miR-252. Overall, our data support a role for the ecdysone/miR-252/mbt regulatory axis in growth control during Drosophila development.


Who's in and who's out: use of primary medical care among people living with HIV.
We sought to achieve a more comprehensive picture of access to medical care for the treatment of HIV (HIV primary care) by combining evidence of medical services used (health utilization) and epidemiological client-level data. This study integrated health information from several data sources to measure utilization of HIV primary care in the St Louis, Mo, area between 1998 and 2002. Data from disparate HIV health utilization databases were combined with data from the Missouri HIV and AIDS Reporting System database and then deidentified to measure client-level utilization of HIV primary care over time. About half of those with HIV showed evidence of having utilized HIV primary care in a given year. Although most of this group utilized HIV primary care in the first year after they received their HIV diagnosis, evidence of subsequent utilization was inconsistent. Utilization of primary care was most strongly associated with an AIDS diagnosis. About one quarter of people diagnosed with HIV after 1997 had an AIDS diagnosis when they first tested positive for HIV. This study was the first of its kind to integrate several databases to understand HIV primary health care utilization over a period of years. Our findings reinforce the importance of CD4 and viral load values as indicators of utilization of HIV primary health care, particularly in the absence of other health data sets. The lack of available data and the way in which source data were available limited the study.


Respiratory muscle strength and the risk of incident cardiovascular events.
Maximal inspiratory pressure (MIP) is a measure of inspiratory muscle strength. The prognostic importance of MIP for cardiovascular events among elderly community dwelling individuals is unknown. Diminished forced vital capacity (FVC) is a risk factor for cardiovascular events which remains largely unexplained. MIP was measured at the baseline examination of the Cardiovascular Health Study. Participants had to be free of prevalent congestive heart failure (CHF), myocardial infarction (MI), and stroke. Subjects in the lowest quintile of MIP had a 1.5-fold increased risk of MI (HR 1.48, 95% CI 1.07 to 2.06) and cardiovascular disease (CVD) death (HR 1.54, 95% CI 1.09 to 2.15) after adjustment for non-pulmonary function covariates. There was a potential inverse relationship with stroke (HR 1.36, 95% CI 0.97 to 1.90), but there was little evidence of an association between MIP and CHF (HR 1.22, 95% CI 0.93 to 1.60). The addition of FVC to models attenuated the HR associated with MIP only modestly; similarly, addition of MIP attenuated the HR associated with FVC only modestly. A reduced MIP is an independent risk factor for MI and CVD death, and a suggestion of an increased risk for stroke. This association with MIP appeared to be mediated through mechanisms other than inflammation.


An international evaluation of biosecurity management capacity in the seaweed aquaculture industry.
Seaweeds form the second largest global aquaculture product in volume, and despite rapid growth of the sector over the last 25 years, production and quality in top producing regions is becoming increasingly limited due to disease and pest outbreaks, the spread of non-native cultivars and the degradation of genetic health due to inbreeding. Most notably, the lack of biosecurity measures leading to disease and pest outbreaks are reported to cause the most significant production losses in the seaweed industry. This study uses the Knowledge, Attitude and Practice (KAP) survey tool to quantify and compare biosecurity cross-culturally, in two major red seaweed producing countries, the Philippines and Tanzania. Both countries have significantly different political contexts and the seaweed sector sits within two very different value chains. Seaweed-based commodities from these countries, however, enters the same international market for carrageenan, a thickening agent used for a variety of products globally. This study uses the KAP survey tool to assess currently-adopted biosecurity control measures and understand how potential policy strategies could be developed on an international scale. Farmers from both producing countries have good biosecurity knowledge. In Tanzania 64% farmers scored Fair or Good, and in the Philippines this was 95%. Corresponding scores in practices were lower, 85% Poor for Tanzania, and 88% Fair for the Philippines, indicating there is a lack of resources for farmers to implement additional practices. The information gathered using the KAP tool in the context of the global seaweed industry can be used to facilitate compromise between science, policy and practice whilst taking into consideration smaller-scale regional challenges. Given the results from the seaweed industry were similar to that of smallholder agricultural sectors, it is suggested that governmental programs to incentivise biosecurity in smallholder rural agriculture could be adapted for the seaweed industry. This study also demonstrates the potential use of the KAP survey, as a tool to accurately compare biosecurity challenges faced by farmers in different aquaculture sectors globally, and to encourage alignment in international approaches to aquaculture biosecurity policies.


Outcomes Associated With Norepinephrine Use Among Cardiac Intensive Care Unit Patients with Severe Shock.
Recent data suggests improved outcomes among cardiac intensive care unit (CICU) patients treated with norepinephrine, especially patients with severe shock. We aimed to describe the association between norepinephrine and mortality in CICU patients with severe shock, defined as those requiring high-dose vasopressors (HDV). We retrospectively evaluated Mayo Clinic CICU patients treated with vasopressors from 2007 to 2015. HDV was defined as a peak Cumulative Vasopressor Index of four for any vasopressor. Peak norepinephrine equivalent (NEE) dose was used to compare vasopressor doses. Multivariable logistic regression was used to determine predictors of hospital mortality. We included 2,090 patients with a median age of 69 years (IQR 59-78), including 35% females; 44% of patients received HDV. Hospital mortality was higher among patients receiving HDV (42% vs. 16%, unadjusted OR 3.87, 95% CI 3.16-4.75, P < 0.01). On multivariable analysis in HDV patients, hospital mortality increased with rising peak NEE (adjusted OR 1.02 per 0.01 mcg/kg/min, 95% CI 1.01-1.02, P < 0.01) and the use of NE was associated with lower hospital mortality (adjusted OR 0.46, 95% CI 0.31-0.72, P < 0.01). After adjustment for illness severity, peak NEE and norepinephrine use were not associated with mortality among patients who did not require HDV. Mortality is high among CICU patients requiring HDV, and rises with increasing vasopressor requirements. Use of NE was associated with lower mortality among patients requiring HDV, but not among those without HDV, implying that patients with more severe shock may benefit from preferential use of NE.


Thomas Birch's 'Weekly Letter' (1741-66): correspondence and history in the mid-eighteenth-century Royal Society.
Thomas Birch (1705-66), Secretary of the Royal Society from 1752 to 1765, and Philip Yorke, second Earl of Hardwicke (1720-90), wrote a 'Weekly Letter' from 1741 to 1766, an unpublished correspondence of 680 letters now housed in the British Library (Additional Mss 35396-400). The article examines the dimensions and purposes of this correspondence, an important conduit of information for the influential coterie of the 'Hardwicke circle' gathered around Yorke in the Royal Society. It explores the writers' self-conception of the correspondence, which was expressed in deliberately archaic categories of seventeenth-century news exchange, such as the newsletter, aviso and a-la-main. It shows how the letter writers negotiated their difference in status through the discourse of friendship, and concludes that the 'Weekly Letter' constituted for the correspondents a form of private knowledge, restricted in circulation to their discrete group, and as such unlike the open and networked model of Enlightenment science.


Institutional experience with 500 cases of surgically treated pediatric Chiari malformation Type I.
The diagnosis and treatment of Chiari malformation Type I (CM-I) has evolved over the last few decades. The authors present their surgical experience of over 2 decades of treating children with this form of hindbrain herniation. The authors conducted a retrospective review of their institutional experience with the surgical treatment of the pediatric CM-I from 1989 to 2010. The 2 most common presentations were headache/neck pain (40%) and scoliosis (18%). Common associated diagnoses included neurofibromatosis Type 1 (5%) and idiopathic growth hormone deficiency (4.2%). Spine anomalies included scoliosis (18%), retroversion of the odontoid process (24%), Klippel-Feil anomaly (3%), and atlantooccipital fusion (8%). Approximately 3% of patients had a known family member with CM-I. Hydrocephalus was present in 48 patients (9.6%). Syringomyelia was present in 285 patients (57%), and at operation, 12% of patients with syringomyelia were found to have an arachnoid veil occluding the fourth ventricular outlet. Fifteen patients (3%) have undergone reoperation for continued symptoms or persistent large syringomyelia. The most likely symptoms and signs to resolve following surgery were Valsalva-induced headache and syringomyelia. The average hospital stay and "return to school" time were 3 and 12 days, respectively. The follow-up for this group ranged from 2 months to 15 years (mean 5 years). Complications occurred in 2.4% of cases; there was no mortality. No patient required acute return to the operating room, and no blood transfusions were performed. The authors believe this to be the largest reported series of surgically treated pediatric CM-I patients and hope that their experience will be of use to others who treat this surgical entity.


The link between major risk factors and important categories of admission in an ageing cohort.
Record linkage of routine hospital data to population-based research findings presents an opportunity to explore the relationships between classical risk factors and hospital activity. The objectives of this study were to examine, in Paisley and Renfrew, the effect of risk factor variables on the likelihood of experiencing an acute hospital admission with six major medical conditions. The subjects were 8,349 women and 7,057 men, aged 45-64 in the early to mid-1970s. The main outcome measures were acute hospital admission with principal diagnosis of: any malignant neoplasm; malignant neoplasm of trachea, bronchus and lung; ischaemic heart disease; respiratory disease; cerebrovascular disease; or diabetes mellitus. Smokers were almost eight times more likely to be admitted with lung cancer and, to a lesser extent, were more likely to be admitted for the other conditions investigated with the exception of diabetes mellitus. Forced expiratory volume was also an independent risk factor for admission with lung cancer and strokes. Higher levels of cholesterol were associated with increased risk of admission with ischaemic heart disease but less with cancer (including lung cancer). With the exception of admissions for cerebrovascular disease, deprivation category was found to have no independent effect on the likelihood of experiencing any of the morbidity outcomes examined. These data confirm that associations first established between risk factors and mortality outcomes (e.g. smoking and lung cancer) are also found between risk factors and hospital admissions for the same causes. This in itself is unremarkable, but the results are of interest for three reasons. First, they illustrate the potential of record linkage to map the effects of risk factors. Second, they demonstrate the size of the effect risk factors have on the risk of admission. Third, they provide a surprising finding that deprivation category does not act as an independent risk factor for the majority of the categories of admission investigated.


Reduced sensitivity to visual looming inflates the risk posed by speeding vehicles when children try to cross the road.
Almost all locomotor animals respond to visual looming or to discrete changes in optical size. The need to detect and process looming remains critically important for humans in everyday life. Road traffic statistics confirm that children up to 15 years old are overrepresented in pedestrian casualties. We demonstrate that, for a given pedestrian crossing time, vehicles traveling faster loom less than slower vehicles, which creates a dangerous illusion in which faster vehicles may be perceived as not approaching. Our results from perceptual tests of looming thresholds show strong developmental trends in sensitivity, such that children may not be able to detect vehicles approaching at speeds in excess of 20 mph. This creates a risk of injudicious road crossing in urban settings when traffic speeds are higher than 20 mph. The risk is exacerbated because vehicles moving faster than this speed are more likely to result in pedestrian fatalities.


IL-10-Secreting CD8+ T Cells Specific for Human Cytomegalovirus (HCMV): Generation, Maintenance and Phenotype.
HCMV-specific CD8+ T-cells are potent anti-viral effector cells in HCMV infected individuals, but evidence from other viral infections suggests that CD8+ T-cells can also produce the immunomodulatory cytokine IL-10. In this work we show that there are HCMV-specific IL-10 CD8+ T-cell responses in a cohort of individuals aged 23-76 years of age, predominantly directed against the HCMV proteins known to be expressed during latent infections as well as towards the proteins US3 and pp71. The analysis of HCMV-specific responses established during primary infection has shown that the IL-10 responses to US3 and pp71 HCMV proteins are detectable in the first weeks post infection, but not the responses to latency-associated proteins, and this IL-10 response is produced by both CD8+ and CD4+ T-cells. Phenotyping studies of HCMV-specific IL-10+ CD8+ T-cells show that these are CD45RA+ effector memory cells and co-express CD28 and CD57, however, the expression of the inhibitory receptor PD-1 varied from 90% to 30% between donors. In this study we have described for the first time the HCMV-specific IL-10 CD8+ T-cell responses and have demonstrated their broad specificity and the potential immune modulatory role of the immune response to HCMV latent carriage and periodic reactivation.


Systemic corticosteroids for the prevention of bronchopulmonary dysplasia, a network meta-analysis.
Despite considerable improvement in outcomes for preterm infants, rates of bronchopulmonary dysplasia (BPD) remain high, affecting an estimated 33% of very low birthweight infants, with corresponding long-term respiratory and neurosensory issues. Systemic corticosteroids can address the inflammation underlying BPD, but the optimal regimen for prevention of this disease, balancing of the benefits with the potentially meaningful risks of systemic corticosteroids, continues to be a medical quandary. Numerous studies have shown that systemic corticosteroids, particularly dexamethasone and hydrocortisone, effectively treat or prevent BPD. However, concerning short and long-term side effects have been reported and the optimal approach to corticosteroid treatment remains unclear. To determine whether differences in efficacy and safety exist between high-dose dexamethasone, moderate-dose dexamethasone, low-dose dexamethasone, hydrocortisone, and placebo in the prevention of BPD, death, the composite outcome of death or BPD, and other relevant morbidities, in preterm infants through a network meta-analysis, generating both pairwise comparisons between all treatments and rankings of the treatments. We searched the Cochrane Library for all systematic reviews of systemic corticosteroids for the prevention of BPD and searched for completed and ongoing studies in the following databases in January 2023: Cochrane Central Register of Controlled Trials, MEDLINE, Embase, and clinical trial databases. We included randomized controlled trials (RCTs) in preterm infants (< 37 weeks' gestation) at risk for BPD that evaluated systemic corticosteroids (high-dose [≥ 4 mg/kg cumulative dose] dexamethasone, moderate-dose [≥ 2 to < 4 mg/kg] dexamethasone, low-dose [< 2 mg/kg] dexamethasone, or hydrocortisone) versus control or another systemic corticosteroid. Our main information sources were the systematic reviews, with reference to the original manuscript only for data not included in these reviews. Teams of two paired review authors independently performed data extraction, with disagreements resolved by discussion. Data were entered into Review Manager 5 and exported to R software for network meta-analysis (NMA). NMA was performed using a frequentist model with random-effects. Two separate networks were constructed, one for early (< seven days) initiation of treatment and one for late (≥ seven days) treatment initiation, to reflect the different patient populations evaluated. We assessed the certainty of evidence derived from the NMA for our primary outcomes using principles of the GRADE framework modified for application to NMA. We included 59 studies, involving 6441 infants, in our analyses. Only six of the included studies provided direct comparisons between any of the treatment (dexamethasone or hydrocortisone) groups, forcing network comparisons between treatments to rely heavily on indirect evidence through comparisons with placebo/no treatment groups. Thirty-one studies evaluated early corticosteroid treatment, 27 evaluated late corticosteroid treatment, and one study evaluated both early and late corticosteroid treatments. Early treatment (prior to seven days after birth): Benefits:NMA for early treatment showed only moderate-dose dexamethasone to decrease the risk of BPD at 36 weeks' postmenstrual age (PMA) compared with control (RR 0.56, 95% CI 0.39 to 0.80; moderate-certainty evidence), although the other dexamethasone dosing regimens may have similar effects compared with control (high-dose dexamethasone, RR 0.71, 95% CI 0.50 to 1.01; low-certainty evidence; low-dose dexamethasone, RR 0.83, 95% CI 0.67 to 1.03; low-certainty evidence). Other early treatment regimens may have little or no effect on the risk of death at 36 weeks' PMA. Only moderate-dose dexamethasone decreased the composite outcome of death or BPD at 36 weeks' PMA compared with control (RR 0.77, 95% CI 0.60 to 0.98; moderate-certainty evidence). Low-dose dexamethasone increased the risk for cerebral palsy (RR 1.92, 95% CI 1.12 to 3.28; moderate-certainty evidence) compared with control. Hydrocortisone may decrease the risk of major neurosensory disability versus low-dose dexamethasone (RR 0.65, 95% CI 0.41 to 1.01; low-certainty evidence). Late treatment (at seven days or later after birth): Benefits: NMA for late treatment showed high-dose dexamethasone to decrease the risk of BPD both versus hydrocortisone (RR 0.66, 95% CI 0.51 to 0.85; low-certainty evidence) and versus control (RR 0.72, CI 0.59 to 0.87; moderate-certainty evidence). The late treatment regimens evaluated may have little or no effect on the risk of death at 36 weeks' PMA. High-dose dexamethasone decreased risk for the composite outcome of death or BPD compared with all other treatments (control, RR 0.69, 95% CI 0.59 to 0.80, high-certainty evidence; hydrocortisone, RR 0.69, 95% CI 0.58 to 0.84, low-certainty evidence; low-dose dexamethasone, RR 0.73, 95% CI 0.60 to 0.88, low-certainty evidence; moderate-dose dexamethasone, RR 0.76, 95% CI 0.62 to 0.93, low-certainty evidence). No effect was observed for the outcomes of major neurosensory disability or cerebral palsy. The evidence for the primary outcomes was of overall low certainty, with notable deductions for imprecision and heterogeneity across the networks. While early treatment with moderate-dose dexamethasone or late treatment with high-dose dexamethasone may lead to the best effects for survival without BPD, the certainty of the evidence is low. There is insufficient evidence to guide this therapy with regard to plausible adverse long-term outcomes. Further RCTs with direct comparisons between systemic corticosteroid treatments are needed to determine the optimal treatment approach, and these studies should be adequately powered to evaluate survival without major neurosensory disability.


Thermometry during MR-guided focused ultrasound in a preclinical model based on Thiel embalmed tissue.
The purpose of this work was to determine the accuracy of Proton Resonance Frequency (PRF) thermometry during MR-guided Focused Ultrasound (MRgFUS) ablation on explanted Thiel embalmed human and animal liver, fresh animal liver, and compared to gel phantom. PRF thermometry during MRgFUS was conducted using a 1.5T MRI system. The phantom and the organs were sonicated with the following energies: 300J, 600J, 1000J and 1400J. The temperature increase which was measured using PRF thermometry during sonication was compared to actual temperature rise in the same conditions measured by fibre optic thermocouple. Sonication of fresh animal liver showed temperature differences varying between 0.27°C and 0.40°C, whereas the phantom results showed temperature differences from 0.23°C to 0.40°C. For the Thiel embalmed organs, the temperature difference varied from 1.17 °C to 3.13°C for the ovine liver, and from 1.3°C to 3.10°C for the human liver. The temperature differences measured in the fresh liver were small and similar to those found for the gel phantom. However, the temperature differences calculated for the Thiel embalmed organs were higher compared to the fresh organ. This indicates that the PRF-based temperature calibration of the Focused Ultrasound machine for Thiel embalmed tissue is necessary.


Intracranial extra-axial ependymoma involving the petroclival region: a rare case report.
Intracranial extraaxial ependymomas (IEAEs) are very rare, and extra-axial petroclival ependymoma (EAPE) has not been reported in the reviewed English-language literature by now. We present a male patient with EAPE misdiagnosed as a petroclival meningioma preoperatively, and involved recurrence at 4th months after operation though totally resection. Thereafter, we report this case and discuss the clinical characteristics of the disease by briefly review.


Regulatory T Cell Immunity in Atherosclerosis.
Atherosclerosis is a chronic inflammatory disorder involving innate and adaptive immunity process. Effector T cell (Teff) responses promote atherosclerotic disease, whereas regulatory T cells (Tregs) have been shown to play a protective role against atherosclerosis by down-regulating inflammatory responses which include multiple mechanisms. Compelling experimental data suggest that shifting the Treg/Teff balance toward Tregs may be a possible therapeutic approach for atherosclerotic disease, although the role of Tregs in human atherosclerotic disease has not been fully elucidated. In this review, we discuss recent advances in our understanding of the roles of Tregs and Teffs in experimental atherosclerosis, as well as human coronary artery disease.


Neuronavigation-assisted single transseptal catheter implantation and shunt in patients with posthemorrhagic hydrocephalus and accentuated lateral ventricles dilatation.
To assess the treatment of posthemorrhagic hydrocephalus with accentuated lateral ventricles dilatation by employing a single biventricular neuronavigation-assisted transseptal-implanted catheter with programmable valve and distal peritoneal derivation. A neuronavigation-assisted single transseptal biventricular catheter implantation with distal peritoneal shunt system was performed in 11 patients with posthemorrhagic hydrocephalus and accentuated lateral ventricles dilatations between 2001 and 2010. Patients with concomitant third ventricle dilatation were excluded. Several sequential frustrated attempts of temporary drainage occlusion on both sides confirmed the isolation of the lateral ventricles. Neuronavigation was employed to accurately establish the catheter surgical corridor (trajectory) across the lateral ventricles and throughout the septum pellucidum. The neurological and radiological outcomes were assessed at least 6 months after the procedure. Catheter implantation was successfully performed in all patients. Only one catheter was found to be monoventricular on delayed computer tomography controls. Procedure-related complications (bleeding of infections) were not observed. No additional neurological deficits were found after shunt surgery. Six months after procedure, none required additional ventricular catheter implantations or shunt revisions. Radiological and clinical controls confirmed the shunt function and the improved neurological status of all patients. Single neuronavigation-assisted transseptal-implanted biventricular catheter is a valid option for the treatment of posthemorrhagic hydrocephalus with accentuated lateral ventricles dilatation. This technique reduces the number of catheters and minimizes the complexity and timing of the surgical procedure as well as potential infection's risks associated with the use of multiple shunting systems.


No evidence of ARAF, CRAF and MET mutations in BRAFT1799A negative human papillary thyroid carcinoma.
The MAPK signaling pathway plays a crucial role in tumorgenesis and cell proliferation in human papillary thyroid carcinoma (PTC). Ret/PTC rearrangements, RAS and BRAF mutations, the main non-overlapping genetic alterations all leading to MAPK cascade activation, are cumulatively identified in 60-80% of PTCs. In approximately one-fourth of the cases, oncogenic background potentially contributing to MAPK activation in PTC might be different. We therefore attempted to evaluate the mutational status of genes encoding other members of RAF family known to act upstream of MAPKs, ARAF and CRAF (RAF-1). In addition we also analyzed the MET gene that encodes hepatocyte growth factor/scatter factor receptor overexpressed in most of PTCs and a MAPK cascade contributor. In 129 Japanese patients with PTC, BRAF(T1799A) was detected in 65 cases (50.4%), and the remaining 64 tumor specimens were subjected to mutation analysis of kinase domains of ARAF, CRAF and MET genes, and hotspots of K- and N-RAS genes. No ARAF, CRAF, MET, K- and N-RAS mutations were revealed. Based on these observations, we concluded that despite the fact that ARAF, CRAF and MET are actively expressed, alterations of these genes are rare in PTC and unlikely to play a perceptible role in the molecular pathogenesis of this type of human malignancy.


[Medical problems and risks of switching drugs according to legal requirements of drug discount contracts in Germany].
Since 2007 German health insurance funds may conclude discount contracts with pharmaceutical companies for individual drugs. According to German legislation pharmacies are liable to preferentially dispense these drugs to patients of the respective funds if the prescribed drug is identical regarding active ingredient, strength, package size, and route of administration, and is approved for the same indication. We aimed to assess the number and nature of clinically relevant differences between prescribed drug and its legal alternatives. Using databases and expert systems of the drug information system AiD KLINIK we evaluated all 258 412 exchangeable drug pairs of the German market currently regulated by discount contracts. 15,7 % of the drug pairs differed in dosage, in one quarter each colour or shape was significantly different, and in roughly 3 % the size of the substituted drug differed by more than 50 %. In 9,43 % splitting characteristics of solid oral doses differed and in 1,87 % the substituted drug contained additives with allergenic potential not present in the primarily selected drug. On average 0,44 clinically relevant differences could be calculated in each substitution. This study has revealed that current legal provision ignore important medical criteria of the substitution process including individual risks (e. g. allergies). Patients will have to change the drug application process and will therefore need appropriate information and training. All these differences between substitutional drug pairs should already be known when prescribing so as to maintain patient safety in the face of this merely cost-saving measure.


FDA Approval Summary: Pirtobrutinib for Relapsed or Refractory Mantle Cell Lymphoma.
In January 2023, the U.S. Food and Drug Administration granted accelerated approval to pirtobrutinib for the treatment of adult patients with relapsed or refractory mantle cell lymphoma (MCL) after at least two lines of systemic therapy, including a Bruton tyrosine kinase (BTK) inhibitor. Approval was based on BRUIN, a single-arm study of pirtobrutinib monotherapy in patients with B-cell malignancies. Efficacy was based on independent review committee- assessed overall response rate (ORR) supported by durability of response in 120 patients with relapsed or refractory MCL who had received a prior BTK inhibitor and received the approved pirtobrutinib dosage of 200 mg once daily. The ORR was 50% (95% confidence interval [CI]: 41, 59) and the complete response rate was 13% (95% CI: 7, 20), with an estimated median duration of response of 8.3 months. The most common non-hematologic adverse reactions were fatigue, musculoskeletal pain, diarrhea, edema, dyspnea, pneumonia, and bruising. Warnings and Precautions in labeling include infection, hemorrhage, cytopenias, atrial arrhythmias, and second primary malignancies. Postmarketing studies were required to evaluate longer-term safety of pirtobrutinib and to verify the clinical benefit of pirtobrutinib. This article summarizes key aspects of the regulatory review, including the indication statement, efficacy and safety considerations, and postmarketing requirements.


Larvicidal Activities of Allium sativum L. and Zingiber officinale Rosc. Extracts against Filariasis Vectors in Hadiya Zone, Ethiopia.
Mosquitoes present an immense threat to millions of people worldwide and act as vectors for filariasis disease. The objective of the study was to determine the effect of Allium sativum and Zingiber officinale extracts against filariasis vectors. The larvae were collected from the breeding site by using standard procedures for identification and larvicidal activities. Twenty grams (20 g) from each (Allium sativum and Zingiber officinale) were extracted separately by aqueous, ethanol, and methanol solvents. The phytochemical analysis was determined in the crude sample by using standard methods. Then, larvicidal effects were determined by introducing 10 larvae of the vectors to the concentrations of 250 ppm, 500 ppm, and 750 ppm of the crude sample, and data were subjected to probit analysis to determine the LC50 and Chi-squared test to check the significance of the mortality by R software. Anopheles funestus, Anopheles gambiae s.l., Anopheles pharoensis, Culex antennatus, and Culex quinquefasciatus were the filariasis vectors identified during the study period. The presence of phytochemical tests such as anthraquinones, flavonoids, glycosides, phenol, saponin, steroids, tannin, and terpenes was obtained. The larvicidal effects of the selected plant extracts ranged from 0%-100%. The lowest LC50 (53 ppm) was observed for A. sativum methanol test extract against Cx. quinquefasciatus. Ethanol extracts of A. sativum have a significant effect on An. funestus (X2 = 7.5, p = 0.02352) and Cx. quinquefasciatus (X2 = 10.833, p = 0.0.0044), whereas aqueous extracts have a significant effect only on An. gambiae s.l. (X2 = 7.0807, p = 0.029. Ethanol extracts of Z. officinale have a significant effect only on the mortality of An. pharoensis (X2 = 7.0807, p = 0.029), but methanol and aqueous extracts have no significant effect against filariasis vectors. In conclusion, A. sativum have a high toxic effect than Z. officinale extract against filariasis vectors in all type of solvents. So using those plant extracts is the best to reduce the risk of the synthetic chemical on nontarget organisms and the environment, in addition to the control of mosquito-borne diseases, but further studies will be conducted to evaluate the toxicity at different stages of the vectors.


Construction of the seed-coat cDNA microarray and screening of differentially expressed genes in barley.
Some barley mutants can synthesize neither anthocyanins nor proanthocyanidins in the seed coat, which is related to several genes in locus Ant13, but the exact model of action remains unknown. We used the cDNA microarray technology with barley transcription-deficient mutant (ant13-152) that does not synthesize proanthocyanidins as the tester, and its wild type genotype (Triumph) as the driver, to study this question. Six-thousand and forty-eight clones from the wild type Morex testa+pericarp cDNA library were amplified using PCR, and the DNA fragments were spotted on commercial amino-modified glass slide as microarray. The mRNAs from the developing seed coat (8-15 days) of both the mutant and the wild-type barley plants were isolated, and labeled respectively with Cy3-dUTP and Cy5-dUTP when reversely transcribed to cDNAs. The labeled cDNAs were used as probes, mixed at the same molar concentration, and hybridized with the DNA fragments on the slide. Seventy clones exhibiting marked differential expression (ratio>4) were identified from the microarray. All the 25 cDNA clones that showed an over-expression in wild type in comparison to the mutant ant13-152 were sequenced. It was found that most of these overexpressing clones were transcription/translation and hordein-associated genes. These results have laid a solid material basis for further elucidation of the metabolic pathway in proanthocyanidin synthesis in barley and likely other plants.


Carcinogenesis of urethane: simulation versus experiment.
The carcinogenesis of urethane (ethyl carbamate), a byproduct of fermentation that is consistently found in various food products, was investigated with a combination of kinetic experiments and quantum chemical calculations. The main objective of the study was to find ΔG(⧧), the activation free energy for the rate-limiting step of the SN2 reaction among the ultimate carcinogen of urethane, vinyl carbamate epoxide (VCE), and different nucleobases of the DNA. In the experimental part, the second-order reaction rate constants for the formation of the main 7-(2-oxoethyl)guanine adduct in aqueous solutions of deoxyguanosine and in DNA were determined. A series of ab initio, density functional theory (DFT), and semiempirical molecular orbital (MO) calculations was then performed to determine the activation barriers for the reaction between VCE and nucleobases methylguanine, methyladenine, and methylcytosine. Effects of hydration were incorporated with the use of the solvent reaction field method of Tomasi and co-workers and the Langevine dipoles model of Florian and Warshel. The computational results for the main adduct were found to be in good agreement with the experiment, thus presenting strong evidence for the validity of the proposed SN2 mechanism. This allowed us to predict the activation barriers of reactions leading to side products for which kinetic experiments have not yet been performed. Our calculations have shown that the main 7-(2-oxoethyl)deoxyguanosine adduct indeed forms preferentially because the emergence of other adducts either proceeds across a significantly higher activation barrier or the geometry of the reaction requires the Watson-Crick pairs of the DNA to be broken. The computational study also considered the questions of stereoselectivity, the ease of the elimination of the leaving group, and the relative contributions of the two possible reaction paths for the formation of the 1,N(2)-ethenodeoxyguanosine adduct.


Towards next-generation model microorganism chassis for biomanufacturing.
Synthetic biology provides powerful tools and novel strategies for designing and modifying microorganisms to function as cell factories for biomanufacturing, which is a promising approach for realizing chemical production in a green and sustainable manner. Recent advances in genetic component design and genome engineering have enabled significant progresses in the field of synthetic biology chassis that have been developed for enzymes or biochemical production based on synthetic biology strategies, with particular reference to model microorganisms, such as Escherichia coli, Bacillus subtilis, Corynebacterium glutamicum, and Saccharomyces cerevisiae. In this review, strategies for engineering four different functional cellular modules which encompass the total process of biomanufacturing are discussed, including expanding the substrate spectrum for substrate uptake modules, refactoring biosynthetic pathways and dynamic regulation for product synthesis modules, balancing energy and redox modules, and cell membrane and cell wall engineering of product storage and secretion modules. Novel strategies of integrating and coordinating different cellular modules aided by synthetic co-culturing of multiple chassis, artificial intelligence-aided data mining for guiding strain development, and the process for designing automatic chassis development via biofoundry are expected to generate next generations of model microorganism chassis for more efficient biomanufacturing. KEY POINTS: • Engineering of functional cellular modules facilitate next generations of chassis construction. • Global optimization of biosynthesis can be improved by metabolic models. • Data-driven and automatic strain development can improve microorganism chassis construction.


Ventral medial prefrontal cortex and emotional perseveration: the memory for prior extinction training.
Several years ago, we found that lesions of ventral medial prefrontal cortex (mPFCv) disrupted performance during the extinction component of a classical fear conditioning task without affecting acquisition performance. We called this emotional perseveration, hypothesizing that mPFCv may normally act to inhibit fear responses to a conditioned stimulus (CS) when the CS no longer signals danger. Subsequent studies have supported this hypothesis, showing that mPFCv is crucial for the memory of prior extinction training. The present study examined the effects of mPFCv lesions made after training. Such lesions resulted in reduced freezing to contextual stimuli and normal responding to the CS presented alone during a retention test. Rats were then subjected to extinction trials (CS without US) over multiple days. In contrast to pre-training lesions, post-training lesions had little effect on extinction rate. All rats were given additional training. Lesioned rats expressed greater fear reactions than controls, indicating that prior extinction was less effective in them. Lesioned rats also showed resistance to extinction during reextinction trials, confirming our earlier finding that lesions made before training weaken the effectiveness of extinction trials. These results suggest three conclusions. First, an intact mPFCv during acquisition may protect the animal from prolonged responding during extinction trials following brain insult. Second, changes in mPFCv may predispose subjects toward enhanced fear reactions that are difficult to extinguish when reexposed to fearful stimuli, due to a diminished capacity to benefit from the fear-reducing impact of prior extinction experience. Third, contextual cues processed by mPFCv may influence extinction performance.


Abnormal electrophysiological phenotypes and sleep deficits in a mouse model of Angelman Syndrome.
Angelman Syndrome (AS) is a rare genetic disorder characterized by impaired communication, motor and balance deficits, intellectual disabilities, recurring seizures and abnormal sleep patterns. The genetic cause of AS is neuronal-specific loss of expression of UBE3A (ubiquitin-protein ligase E6-AP), an imprinted gene. Seizure and sleep disorders are highly prevalent (> 80%) in the AS population. The present experiments were designed to identify translational, neurophysiological outcome measures in a model of AS. We used the exon-2 deletion mouse (Ube3a-del) on a C57BL/6J background to assess seizure, sleep and electrophysiological phenotypes. Seizure susceptibility has been reported in Ube3a-del mice with a variety of seizure induction methods. Here, we provoked seizures by a single high-dose injection of 80 mg/kg pentylenetetrazole. Novel experiments included the utilization of wireless telemetry devices to acquire global electroencephalogram (EEG) and neurophysiological data on electrographic seizures, power spectra, light-dark cycles, sleep stages and sleep spindles in Ube3a-del and WT mice. Ube3a-del mice exhibited reduced seizure threshold compared to WT. EEG illustrated that Ube3a-del mice had increased epileptiform spiking activity and delta power, which corroborates findings from other laboratories and recapitulates clinical reports in AS. This is the first report to use a cortical surface-based recording by a wireless telemetry device over tethered/fixed head-mount depth recordings. Less time in both paradoxical and slow-wave sleep, longer latencies to paradoxical sleep stages and total less sleep time in Ube3a-del mice were observed compared to WT. For the first time, we detected fewer sleep spindles in the AS mouse model. This study was limited to the exon 2 deletion mouse model, and future work will investigate the rat model of AS, containing a complete Ube3a deletion and pair EEG with behavior. Our data enhance rigor and translatability as our study provides important corroboration of previous reports on epileptiform and elevated delta power. For the first time we report neurophysiological phenotypes collected via translational methodology. Furthermore, this is the first report of reduced sleep spindles, a critical marker of memory consolidation during sleep, in an AS model. Our results are useful outcomes for therapeutic testing.


Presenilin 1 interaction in the brain with a novel member of the Armadillo family.
One approach to understanding the function of presenilin 1 (PS1), is to discover those proteins with which it interacts. Evidence for a function in developmental patterning came from C. elegans, in which a PS homologue was identified by screening for suppressors of a mutation in Notch/lin-12, a gene which specifies cell fate. However, this genetic experiment cannot determine which proteins directly interact with PS1. Therefore, we utilized the two hybrid system and confirmatory co-immunoprecipitations to identify a novel catenin, termed gamma-catenin, which interacts with PS1 and is principally expressed in brain. The catenins are a gene family related to the Armadillo gene in Drosophila, some of which appear to have dual roles-they are components of cell-cell adherens junctions, and may serve as intermediates in the Wingless (Wg) signaling pathway, which, like Notch/lin-12, is also responsible for a variety of inductive signaling events. In the non-neuronal 293 cell line, PS1 interacted with gamma-catenin, the family member with the greatest homology to Armadillo. Wg and Notch interactions are mediated by the Disheveled gene, which may form a signaling complex with PS1 and Wg pathway intermediates to regulate the function of the Notch/lin-12 gene.


Cytokine gene therapy of gliomas: induction of reactive CD4+ T cells by interleukin-4-transfected 9L gliosarcoma is essential for protective immunity.
Tumor cells genetically modified to secrete cytokines stimulate potent immune responses against peripheral and central nervous system tumors; however, variable results on the efficacy of this strategy for therapeutic intervention against established intracranial neoplasia have been reported. We have found that vaccination with rat 9L gliosarcoma cells expressing interleukin 4 (9LmIL4) induced a specific, protective, immune response against rechallenge with parental 9L tumors. In naive rats, sham-transfected 9L (9Lneo) tumors and 9LmIL4 tumors grew at comparable rates for 12-14 days, and then 9LmIL4 tumors regressed. After regression of 9LmIL4 tumors, rats were resistant to rechallenge with parental 9L cells. To investigate the mechanism(s) responsible for 9LmIL4-induced immunity, the phenotype and function of tumor-infiltrating lymphocytes (TILs) in 9Lneo and 9LmIL4 tumors were compared. In flow cytometric analyses, it was determined that CD4+ T cells were the predominant cell type in both 9Lneo and 9LmIL4 tumors at day 10. However, at the onset of regression (day 14), 9LmIL4 tumors were infiltrated predominantly by CD8+ T cells. To investigate functional aspects of the anti-9L tumor responses, we assessed the capacity of 9LmIL4 TILs to mediate specific lytic function or production of cytokines. In response to parental 9L, TILs isolated from day 14 9LmIL4 tumors were demonstrated to produce substantially greater amounts of IFN-gamma than did TILs from 9Lneo tumors. Although freshly isolated TILs from 9LmIL4 or control tumors did not lyse 9L cells in 51Cr-release cytotoxicity assays, specific cytotoxicity was demonstrable using TILs from day 14 9LmIL4 or splenocytes from 9LmIL4-bearing rats after their restimulation for 5 days with parental 9L tumor cells in vitro. Antibody blocking studies demonstrated that cytokine production and lytic activity by TILs, or splenocytes from 9LmIL4-immunized rats, were mediated in a T-cell receptor-dependent fashion. Because interleukin-4 also promotes humoral responses, quantity and isotype of immunoglobulins in sera from 9Lneo or 9LmIL4-immunized rats were compared. The amount of IgG1 antibodies was significantly increased in sera from 9LmIL4-immunized rats compared to sera from 9Lneo-bearing rats. Experiments using sublethally irradiated, naive rats adoptively transferred with splenocytes and/or sera from 9LmIL4-immunized or naive rats demonstrated that immune cells, with or without immune sera, protected recipients from challenge with parental 9L. Immune sera provided no protection when given with lymphocytes from naive rats, and it did not enhance protection against parental 9L when given in conjunction with lymphocytes for 9LmIL4-immunized rats. In additional adoptive transfer experiments, an essential role for CD4+ T cells in immunity was observed because their depletion from among splenocytes of 9LmIL4-immunized rats eliminated the protective effective against 9L, whereas depletion of CD8+ cells resulted in a more limited effect on protection against 9L. These data suggest that strategies for inducing systemic, long-term tumor-specific reactivity among CD4+ T cells will be critical for the development of immunotherapy of gliomas.


SURVIVAL ANALYSIS OF DIALYSIS PATIENTS IN SELECTED HOSPITALS OF LAHORE CITY.
There are several reasons which are directly or indirectly relate to affect the survival time of End Stage Renal Disease (ESRD) patients. This study was done to analyse the survival rate of ESRD patients in Lahore city, and to evaluate the influence of various risk factors and prognostic factors on survival of these patients. A sample of 40 patients was taken from the Jinnah Hospital Lahore and Lahore General Hospital by using the convenience sampling technique. The Log Rank Test was used to determine the significant difference between the categories of qualitative variables of ESRD patients. Multivariate Cox Regression Analysis was used to analyse the effect of different clinical and socio-economic variables on the survival time of these patients. Different qualitative variables like: age, marital status, BMI, comorbid factors, diabetes type, gender, income level, place, risk factor like diabetes, ischemic heart disease, hypertension and Hepatitis status were analysed on the basis of Log Rank Test. While age and comorbid factors were found to be statistically significant which showed that the distribution of age and comorbid factors were different. By using the Cox Regression analysis the coefficient of Mass, serum albumin and family history of diabetes were found to be significant. There were some of the factors which had been taken for the analysis came out less or more significant in patients of ESRD. So it was concluded that mostly clinical factors which were Mass of the Patient, Serum Albumin and Family History of Diabetes made significant contribution towards the survival status of patients.


Alternative processing of androgen-binding protein RNA transcripts in fetal rat liver. Identification of a transcript formed by trans splicing.
Androgens and their nuclear receptor regulate genes necessary for development of the male phenotype, a process that is thought to be modulated by extracellular androgen carrier proteins. Two of these carrier proteins, testicular androgen-binding protein (ABP) and plasma sex hormone-binding globulin (SHBG), are encoded by the same gene, but differ in glycosylation and possibly amino acid sequence. To investigate ABP-SHBG gene expression in fetal rat liver, we analyzed RNA transcripts and expressed protein. These studies demonstrated a transient expression of ABP in hepatocytes during the time of testosterone-dependent differentiation of the Wolffian duct. Analysis of cDNA clones derived from fetal rat liver cDNA libraries identified two cDNAs encoded by the ABP-SHBG gene that represented alternatively spliced RNAs. One cDNA had an alternate exon 1, suggesting the function of another promoter in fetal liver. This cDNA also lacked testicular exon 6 DNA, an alteration that implicates the encoded protein in regulatory functions. The other cDNA represented a fused transcript of the ABP-SHBG gene (exons 1-5) and the histidine decarboxylase (HDC) gene, encoding a Mr 98,000 precursor protein. The two domains were joined at splice junctions of the ABP-SHBG and HDC genes, which were localized to rat chromosomes 10 and 3, respectively. Our results indicate that the joining of the two domains was by a trans (donor and acceptor)-splicing mechanism. Data from Northern hybridization experiments suggest the fusion transcript is present in fetal liver RNA. Polymerase chain reaction experiments with fetal liver cDNA further support the existence of an ABP-HDC fusion transcript, as well as the other alternate mRNA. Moreover, a Mr 93,000 immunoreactive protein was transiently expressed in fetal liver during the time of ABP and HDC gene expression. Expression of the fusion cDNA in COS cells yielded HDC activity and the predicted size protein (Mr = 93,000) on Western immunoblots.


Radiation treatment for incompletely resected soft-tissue sarcomas in dogs.
To evaluate efficacy of radiation for treatment of incompletely resected soft-tissue sarcomas in dogs. Prospective serial study. 48 dogs with soft-tissue sarcomas. Tumors were resected to < 3 cm3 prior to radiation. Tumors were treated on alternate days (three 3-Gy fractions/wk) until 21 fractions had been administered. Cobalt 60 radiation was used for all treatments. Five-year survival rate was 76%, and survival rate was not different among tumor types or locations. Four (8%) dogs developed metastases. Eight (17%) dogs had tumor recurrence after radiation. Development of metastases and local recurrence were significantly associated with reduced survival rate. Median survival time in dogs that developed metastases was 250 days. Median disease-free interval for all dogs was 1,082 days. Median time to recurrence was 700 days. Dogs that developed recurrence after a prolonged period responded well to a second surgery. Acute radiation toxicosis was minimal; osteosarcoma developed at the radiation site in 1 dog. An excellent long-term survival rate may be achieved by treating soft-tissue sarcomas in dogs with resection followed by radiation. Amputation is not necessary for long-term control of soft-tissue sarcomas in limbs. Development of metastases and recurrence of local tumors after radiation treatment are associated with decreased survival rate. Acute and delayed radiation toxicosis was minimal with the protocol used in this study.


Health and well-being of secondary school students in New Zealand: trends between 2001, 2007 and 2012.
To describe indicators of health and well-being for New Zealand secondary school students; explore changes between 2001, 2007 and 2012; and compare these findings to international estimates. Three national health and well-being surveys of randomly selected New Zealand secondary school students were conducted. Data are presented as prevalence and variation over time (adjusted odds ratio (aOR)). Comparisons with international estimates were made with subsets of the data. Between 2001 and 2012, students reported reductions in cigarette use (aOR 0.27, 95% confidence interval (CI) 0.23-0.32), alcohol use (aOR 0.39, 95% CI 0.33-0.46), marijuana use (aOR 0.37, 95% CI 0.31-0.43), sexual abuse (aOR 0.52, 95% CI 0.46-0.58), fighting (aOR 0.63, 95% CI 0.55-0.73), seatbelt use (aOR 1.47, 95% CI 1.31-1.65) and risky driving behaviours (aOR 0.39, 95% CI 0.33-0.45). Positive connections to school (perception that the school cares, aOR 1.22, 95% CI 1.10-1.35; liking school, aOR 1.55, 95% CI 1.33-1.82) and family (good family relationship, aOR 1.83, 95% CI 1.70-1.97) also improved. Indicators that did not improve and compared poorly with international estimates were protected sex (condom use at last sexual intercourse, aOR 0.77, 95% CI 0.68-0.87) and healthy life-style (daily physical activity, aOR 0.88, 95% CI 0.78-0.99; overweight/obese, aOR 1.09, 95% CI 0.92-1.31). Exposure to family violence (aOR 1.37, 95% CI 1.11-1.68) and depressive symptoms (aOR 1.03, 95% CI 0.91-1.17) also did not improve. There have been important improvements in the health and well-being of New Zealand adolescents over a relatively short period. These findings demonstrate that population rates of adolescent risk behaviours are amenable to change. Current policy efforts should not lose momentum, while identified priority areas must be adequately resourced to ensure young people have opportunities to thrive now and in the future.


Nasogastric versus nasojejunal tube feeding for severe acute pancreatitis: A systematic review of randomized controlled trials.
Early enteral nutrition through a feeding tube is essential for the management of severe acute pancreatitis (SAP). Nasojejunal tube nutrition has been preferred on the assumption that it provided pancreatic rest in comparison to the nasogastric tube. However, nasojejunal tube placement is complex, may delay feeding initiation and can increase hospital costs. Our aim was to compare the efficacy and safety of enteral feeding with a nasogastric tube versus nasojejunal tube in patients with SAP. We searched four databases (PubMed, Web of Science, Scopus, and Embase) until December 1, 2022. We included randomized controlled trials (RCTs) comparing enteral feeding by nasogastric and nasojejunal tubes in patients with SAP. Primary outcome was all-cause mortality. Secondary outcomes were organ failure, infection, complications, surgical intervention, duration of tube feeding and length of hospital stay. Risk of bias assessment was completed independently by two investigators using the Cochrane RoB 2.0 tool. We performed random effects model meta-analyses using the inverse variance method. Effect measures were reported as relative risks (RR) and their 95% CIs for dichotomous outcomes and mean differences (MD) and their 95% CIs for continuous outcomes. We included four RCTs involving 192 patients with SAP. The mean ages ranged between 36 and 62 years old. There was no significant difference in all-cause mortality between the nasogastric and nasojejunal feeding arms (18/98 vs. 23/93; RR 1.34, 95%CI 0.77-2.30; p=0.30). There were no significant differences in all secondary outcomes between feeding arms. There were three RCTs with some concerns of bias, in the randomization process. In conclusion, in patients with SAP, enteral feeding delivered by nasogastric tube was as efficacious and safe as nasojejunal tube. Further randomized controlled trials with more participants and better design are needed to confirm these findings.


Glutathione deficiency of the Arabidopsis mutant pad2-1 affects oxidative stress-related events, defense gene expression, and the hypersensitive response.
The Arabidopsis (Arabidopsis thaliana) phytoalexin-deficient mutant pad2-1 displays enhanced susceptibility to a broad range of pathogens and herbivorous insects that correlates with deficiencies in the production of camalexin, indole glucosinolates, and salicylic acid (SA). The pad2-1 mutation is localized in the GLUTAMATE-CYSTEINE LIGASE (GCL) gene encoding the first enzyme of glutathione biosynthesis. While pad2-1 glutathione deficiency is not caused by a decrease in GCL transcripts, analysis of GCL protein level revealed that pad2-1 plants contained only 48% of the wild-type protein amount. In contrast to the wild type, the oxidized form of GCL was dominant in pad2-1, suggesting a distinct redox environment. This finding was corroborated by the expression of GRX1-roGFP2, showing that the cytosolic glutathione redox potential was significantly less negative in pad2-1. Analysis of oxidative stress-related gene expression showed a higher transcript accumulation in pad2-1 of GLUTATHIONE REDUCTASE, GLUTATHIONE-S-TRANSFERASE, and RESPIRATORY BURST OXIDASE HOMOLOG D in response to the oomycete Phytophthora brassicae. Interestingly, oligogalacturonide elicitation in pad2-1 revealed a lower plasma membrane depolarization that was found to act upstream of an impaired hydrogen peroxide production. This impaired hydrogen peroxide production was also observed during pathogen infection and correlated with a reduced hypersensitive response in pad2-1. In addition, a lack of pathogen-triggered expression of the ISOCHORISMATE SYNTHASE1 gene, coding for the SA-biosynthetic enzyme isochorismate synthase, was identified as the cause of the SA deficiency in pad2-1. Together, our results indicate that the pad2-1 mutation is related to a decrease in GCL protein and that the resulting glutathione deficiency negatively affects important processes of disease resistance.


The first UK report of a rare Cryptosporidium hominis genetic variant isolated during a complex Scottish swimming pool outbreak.
Cryptosporidium species are responsible for causing the majority of parasite-related gastrointestinal infections in the UK. This report describes an outbreak of 12 laboratory-confirmed cryptosporidiosis cases identified as part of a Scottish swimming pool investigation, with 9 primary and 3 secondary cases occurring over an 8-week period. Molecular speciation was successful for 11/12 cases, which revealed 10 Cryptosporidium hominis cases and 1 Cryptosporidium parvum case. Of the 10 C. hominis cases, further typing identified 7 as being an unusual sub-type, IbA6G3, which is the first description in the UK of this rare variant. The remaining three C. hominis cases were identified as the common IbA10G2 subtype. Following implementation of control measures on two occasions, no further cases were reported. This report highlights the importance of molecular typing to identify and characterize outbreaks, and emphasizes the need to adhere to swimming pool guidance. It also raises awareness of the potential for outbreaks to involve multiple species/sub-types, and emphasizes the importance of strong public health leadership to ensure effective multi-agency investigations and management of outbreaks.


The GATA-Type Transcriptional Factor Are1 Modulates the Expression of Extracellular Proteases and Cellulases in Trichoderma reesei.
Trichoderma reesei is a biotechnologically important filamentous fungus with the remarkable ability to secrete large amounts of enzymes, whose production is strongly affected by both the carbon and nitrogen sources. While the carbon metabolism regulators are extensively studied, the regulation of enzyme production by the nitrogen metabolism regulators is still poorly understood. In this study, the GATA transcription factor Are1, which is an orthologue of the Aspergillus global nitrogen regulator AREA, was identified and characterized for its functions in regulation of both protease and cellulase production in T. reesei. Deletion of the are1 gene abolished the capability to secrete proteases, and complementation of the are1 gene rescued the ability to produce proteases. Quantitative RT-PCR analysis revealed that the transcripts of protease genes apw1 and apw2 were also significantly reduced in the Δare1 strain when grown in the medium with peptone as the nitrogen source. In addition, deletion of are1 resulted in decreased cellulase production in the presence of (NH4)2SO4. Consistent with the reduction of cellulase production, the transcription levels of the major cellulase genes, including cbh1, cbh2, egl1, and egl2, were dramatically decreased in Δare1. Sequence analysis showed that all promoter regions of the tested protease and cellulase genes contain the consensus GATA elements. However, the expression levels of the major cellulase transcription activator Xyr1 and the repressor Cre1 had no significant difference between Δare1 and the parental strain QM9414, indicating that the regulatory mechanism deserves further investigation. Taken together, these results demonstrate the important role of Are1 in the regulation of protease and cellulase production in T. reesei, although these processes depend on the kind of nitrogen sources. The findings in this study contribute to the understanding of the regulation network of carbon and nitrogen sources in filamentous fungi.


EARN: an ensemble machine learning algorithm to predict driver genes in metastatic breast cancer.
Today, there are a lot of markers on the prognosis and diagnosis of complex diseases such as primary breast cancer. However, our understanding of the drivers that influence cancer aggression is limited. In this work, we study somatic mutation data consists of 450 metastatic breast tumor samples from cBio Cancer Genomics Portal. We use four software tools to extract features from this data. Then, an ensemble classifier (EC) learning algorithm called EARN (Ensemble of Artificial Neural Network, Random Forest, and non-linear Support Vector Machine) is proposed to evaluate plausible driver genes for metastatic breast cancer (MBCA). The decision-making strategy for the proposed ensemble machine is based on the aggregation of the predicted scores obtained from individual learning classifiers to be prioritized homo sapiens genes annotated as protein-coding from NCBI. This study is an attempt to focus on the findings in several aspects of MBCA prognosis and diagnosis. First, drivers and passengers predicted by SVM, ANN, RF, and EARN are introduced. Second, biological inferences of predictions are discussed based on gene set enrichment analysis. Third, statistical validation and comparison of all learning methods are performed by some evaluation metrics. Finally, the pathway enrichment analysis (PEA) using ReactomeFIVIz tool (FDR < 0.03) for the top 100 genes predicted by EARN leads us to propose a new gene set panel for MBCA. It includes HDAC3, ABAT, GRIN1, PLCB1, and KPNA2 as well as NCOR1, TBL1XR1, SIRT4, KRAS, CACNA1E, PRKCG, GPS2, SIN3A, ACTB, KDM6B, and PRMT1. Furthermore, we compare results for MBCA to other outputs regarding 983 primary tumor samples of breast invasive carcinoma (BRCA) obtained from the Cancer Genome Atlas (TCGA). The comparison between outputs shows that ROC-AUC reaches 99.24% using EARN for MBCA and 99.79% for BRCA. This statistical result is better than three individual classifiers in each case. This research using an integrative approach assists precision oncologists to design compact targeted panels that eliminate the need for whole-genome/exome sequencing. The schematic representation of the proposed model is presented as the Graphic abstract.


[Excision repair does not protect Bacillus subtilis cells from inactivation by sunlight].
Exponentially growing Bacillus subtilis cells are highly sensitive to inactivating action of sunlight, the strain deficient in excision repair being every more sensitive than the uvr1 mutant. The inactivating effect is connected with the action of irradiation located in the left part of the spectrum (the whole UV region and some zones of the visible one). Increased sensitivity to sunlight disappeared when cells were exposed to sunlight in a liquid medium with casaminoacids (2 g/l). The inactivating effect was probably of photodynamic nature and was caused either by DNA lesions that were not removed by excision repair or by the damage which arose not in DNA.


Health-related quality of life and pain in children and adolescents: a school survey.
Pain problems are common in children and adolescents. Measures of health-related quality of life (HRQoL) can be used to assess children's subjective perspectives of pain experience and its impact on their life. The aims of the study were to describe HRQoL and the prevalence of pain in a nonclinical population of children and adolescents, and to analyze the relationships between HRQoL, pain, sex, and age in a sample of children and adolescents aged 8-18 years. This cross-sectional study involved a cluster sample of 20 randomly selected schools drawn within a region of Norway. The final study sample included 1099 children and adolescents. We measured HRQoL using the generic questionnaire KIDSCREEN-52 and pain using questions from the Lübeck Pain-Screening Questionnaire. Multiple regression was used to analyze relationships between HRQoL and sex, age, and pain. The response rate was 74%. A large percentage of the sample, 60%, reported pain, and girls reported significantly more pain than boys, 76% of the girls in the age group 16-18 years reported pain. The KIDSCREEN-52 scores differed between girls and boys, and on average, girls reported a significantly lower HRQoL than boys on most dimensions. Pain problems were associated with lower HRQoL, and older girls were most impaired by pain. The findings from this study indicate that pain problems are highly prevalent in children, and more prevalent in girls than in boys. HRQoL was impaired for all 10 dimensions of the KIDSCREEN-52 in children with pain. The subscales self-perception, psychological well-being, mood, relationship with parents, and school environment were most affected.


In silico comparative study of the genomic islands of Vibrio cholerae MJ1236 with those of Classical and El Tor N16961 strains of Vibrio cholerae.
The evolution of microbial genomes is greatly influenced by horizontal gene transfer (HGT), where large blocks of horizontally acquired foreign sequences, often encoding virulence determinants, occur in chromosomes of pathogenic bacteria. A program design-island developed in our laboratory was used on three completely sequenced Vibrio cholerae genomes, V. cholerae Classical O395, El Tor N16961 and MJ1236, in order to identify the putative horizontally acquired regions. The putative genomic islands (GIs) were graphically represented and analyzed. The study identified distinct regions in the GIs of V. cholerae MJ1236 which were shared either with the Classical or the El Tor strain of V. cholerae. A cluster comprising of 38 ORFs was common to V. cholerae strains of MJ1236 and Classical O395 but absent in El Tor N16961. About 5% of the predicted GIs of V. cholerae MJ1236 were unique to itself. Among these unique ORFs, a region of mostly hypothetical genes was identified, where the ORFs were present in a large cluster. The results show that the HGT had played a significant role in the evolution and the differentiation of V. cholerae MJ1236.


The tissue localization of Mycoplasma meleagridis in turkey embryos.
Mycoplasma meleagridis appears to localize primarily in the respiratory tissues of the developing turkey embryo. This was determined by the daily collection throughout the incubation period of smears of embryonic tissues from 261 fertile turkey eggs. The tissues were examined using fluorescent antibody procedures and the earliest detection from positive tissues were: day 6, blastoderm (10%); day 8, sinus (44%) and peritoneum (55%); day 19, lung (11%) and trachea (44%); day 23, air sacs (62%).


New neo-clerodane diterpenoid alkaloids from Scutellaria barbata with cytotoxic activities.
Four new neo-clerodane diterpenoid alkaloids, named scutebarbatines I-L (1-4), were isolated from the whole plant of Scutellaria barbata D. DON. Their structures were established on the basis of detailed spectral analyses. In vitro, the four new compounds showed significant cytotoxic activities against three human cancer lines (HONE-1 nasopharyngeal, KB oral epidermoid carcinoma, and HT29 colorectal carcinoma cells), and gave IC(50) values in the range 3.2-8.3 microM.


Solutions of a three-dimensional multi-term fractional anomalous solute transport model for contamination in groundwater.
The study presents a meshless computational approach for simulating the three-dimensional multi-term time-fractional mobile-immobile diffusion equation in the Caputo sense. The methodology combines a stable Crank-Nicolson time-integration scheme with the definition of the Caputo derivative to discretize the problem in the temporal direction. The spatial function derivative is approximated using the inverse multiquadric radial basis function. The solution is approximated on a set of scattered or uniform nodes, resulting in a sparse and well-conditioned coefficient matrix. The study highlights the advantages of meshless method, particularly their simplicity of implementation in higher dimensions. To validate the accuracy and efficacy of the proposed method, we performed numerical simulations and compared them with analytical solutions for various test problems. These simulations were carried out on computational domains of both rectangular and non-rectangular shapes. The research highlights the potential of meshless techniques in solving complex diffusion problems and its successful applications in groundwater contamination and other relevant fields.