Type V Secretion: the Autotransporter and Two-Partner Secretion Pathways.
The autotransporter and two-partner secretion (TPS) pathways are used by E. coli and many other Gram-negative bacteria to delivervirulence factors into the extracellular milieu.Autotransporters arecomprised of an N-terminal extracellular ("passenger") domain and a C-terminal β barrel domain ("β domain") that anchors the protein to the outer membrane and facilitates passenger domain secretion. In the TPS pathway, a secreted polypeptide ("exoprotein") is coordinately expressed with an outer membrane protein that serves as a dedicated transporter. Bothpathways are often grouped together under the heading "type V secretion" because they have many features in common and are used for the secretion of structurally related polypeptides, but it is likely that theyhave distinct evolutionary origins. Although it was proposed many years ago that autotransporterpassenger domains are transported across the outer membrane through a channel formed by the covalently linked β domain, there is increasing evidence that additional factors are involved in the translocation reaction. Furthermore, details of the mechanism of protein secretion through the TPS pathway are only beginning to emerge. In this chapter I discussour current understanding ofboth early and late steps in the biogenesis of polypeptides secreted through type V pathways and current modelsofthe mechanism of secretion.


The KDM Inhibitor GSKJ4 Triggers CREB Downregulation via a Protein Kinase A and Proteasome-Dependent Mechanism in Human Acute Myeloid Leukemia Cells.
Acute myeloid leukemia (AML) is a progressive hematopoietic-derived cancer arising from stepwise genetic mutations of the myeloid lineage. cAMP response element-binding protein (CREB) is a nuclear transcription factor, which plays a key role in the multistep process of leukemogenesis, thus emerging as an attractive potential drug target for AML treatment. Since epigenetic dysregulations, such as DNA methylation, histone modifications, as well as chromatin remodeling, are a frequent occurrence in AML, an increasing and selective number of epi-drugs are emerging as encouraging therapeutic agents. Here, we demonstrate that the histone lysine demethylases (KDMs) JMJD3/UTX inhibitor GSKJ4 results in both proliferation decrease and CREB protein downregulation in AML cells. We found that GSKJ4 clearly decreases CREB protein, but not CREB mRNA levels. By cycloheximide assay, we provide evidence that GSKJ4 reduces CREB protein stability; moreover, proteasome inhibition largely counteracts the GSKJ4-induced CREB downregulation. Very interestingly, a rapid CREB phosphorylation at the Ser133 residue precedes CREB protein decrease in response to GSKJ4 treatment. In addition, protein kinase A (PKA) inhibition, but not extracellular signal-regulated kinase (ERK)1/2 inhibition, almost completely prevents both GSKJ4-induced p-Ser133-CREB phosphorylation and CREB protein downregulation. Overall, our study enforces the evidence regarding CREB as a potential druggable target, identifies the small epigenetic molecule GSKJ4 as an "inhibitor" of CREB, and encourages the design of future GSKJ4-based studies for the development of innovative approaches for AML therapy.


Patterns in abundance and seasonality of insects in the siruvani forest of Western ghats, nilgiri biosphere reserve, southern India.
The seasonal abundance patterns of insects inhabiting the understory vegetation of a mixed deciduous forest were examined with the help of the sweep-net sampling method. During the study period of 2 years, insects were sampled regularly from the understory vegetation of the three selected habitats (moist-deciduous, riverine, and teak plantation) of the mixed deciduous forest. Insect abundance was maximum in the moist-deciduous habitat and minimum in the teak plantation. Generally, insect abundance was the highest during the southwest monsoon in all habitats. The temporal pattern of fluctuations in the insect abundance followed more or less the same pattern in all the three habitats studied. The insect abundance of the understory vegetation varied among the habitats studied, while the pattern of seasonal fluctuations in insect abundance was comparable among habitats. Composition of the insect community also indicated prominent seasonal changes within habitats than interhabitat changes within a season.


The Impact on Urology Residents' Learning of Social Media and Web Technologies after the Pandemic: A Step Forward through the Sharing of Knowledge.
The COVID-19 pandemic has impacted urology residents and their training. However, several new technologies or knowledge platforms as social media (SoMe) and web-based learning solutions have filled this gap. Therefore, we aimed to analyze resident's learning curves of new learning modalities, identify the evidence that is provided in the literature, and evaluate the possible impact of such solutions in the future. We conducted a non-systematic literature search using Medline, PubMed, and Embase. In addition, online resources of national and international urology resident societies were queried. The identified paper described SoMe, webinars, podcasts, pre-recorded surgical videos, educational platforms, and mobile apps in the field of urology that are used to gain access to information, teach and provide feedback to residents, and were used under the conditions of the pandemic. The application of those technologies harbors the risk of mis- and disinformation, but have the potential to provide access to education and validated knowledge, training, and feedback and thereby might democratize training of residents in urology globally.


Sun protection counseling by pediatricians.
We conducted a pilot study in Florida to assess the extent to which caretakers reported sun protection counseling by their physicians. We also sought to determine whether caretakers who reported receiving counseling had more favorable sun protection knowledge, attitudes, and behaviors. We sampled 100 caretakers who were attending a university pediatric clinic in Florida and obtained 77 responses. We estimated the proportion of respondents who reported receiving sun protection counseling and examined sociodemographic predictors of counseling. Thirty percent of caretakers reported having been counseled by their physician. Sunscreen and avoiding peak sun exposure were the most common items discussed. Among those counseled, 42.9% stated that their child's sun protection habits increased as a result of the counseling. Counseled caretakers had greater sun protection knowledge, were more likely to report regular use of sun protection for their child (68.2% vs 40.9%, P < 0.05), and were more likely to report teaching their children about sun protection (90.9% vs 63.3%, P =.02). Physician sun protection counseling, while infrequently reported, was associated with more favorable sun protection knowledge, attitudes, and self-reported behaviors of caretakers and their children.


Etiology and pathogenesis of anterior open bite: a review.
To review the etiology and pathogenesis of anterior open bite malocclusion. Review of literature was affected through Pubmed, Google scholar and Science direct. References identified from articles found from the primary search were also reviewed. Published data on etiology and pathogenesis of anterior open bite over the last five decades (1960-2009) were utilised. Full articles, abstracts and relevant book chapters were read and analysed to determine the relevant material for this article. All relevant articles were reviewed in full and necessary information eextracted as necessary. A clear understanding of the etiology and pathogenesis of anterior open bite is essential in the diagnosis, prevention and management of this malocclusion.


[Gender-specific post-stimulus modulation in the alpha band during visual-spatial attention].
Previous studies of the spontaneous activity of the EEG indicate that women present bigger absolute spectral power in the alpha band than men, but few studies have been done during cognitive activity. To check possible gender differences in the EEG frequency domain during a visuo-spatial attentional task to understand better the temporary dynamics of alpha. A classic oddball paradigm was used whose stimuli (90% standard-10% objectives) were complex. In each block (10 with 100 trials) it was requested to the subjects (20, 10 of each gender) to attend to one of the visual fields (left and right, with random stimuli and identical probability). Power spectral density values were calculated applying the fast Fourier transform in the alpha band (9-11 Hz) in two overlapped windows (sliding windows): of 0-512 ms and of 412-924 ms. The normalized data were analyzed by means of ANOVAs, with the following factors: gender, attention, hemisphere, visual field and electrodes. Different modulations were obtained attending to the gender variable: men showed smaller alpha or bigger suppression of the rhythm in posterior electrodes and women exhibited smaller power in anterior electrodes.


Polyelectrolyte-bridged metal/cotton hierarchical structures for highly durable conductive yarns.
A novel, facile, and versatile approach for preparing highly durable, electrically conductive cotton yarns is reported. Polyelectrolyte brushes, a polymer that covalently tethers one end on a surface, are first grown from cotton surfaces by surface-initiated atomic transfer radical polymerization. Subsequent electroless deposition of metal particles onto the brush-modified cotton yarns yields electrically conductive yarns, which have conductivity as high as approximately 1 S/cm and can be used as electrical wires in wearable, flexible electronic devices. Importantly, the formation of polymer brush-bridged metal/cotton hierarchical structures provides robust mechanical and electrical durability to the yarns under many stretching, bending, rubbing, and washing cycles. With proper selection of metal, the conductivity of the samples remains stable after they are stored in air for a few months. This chemical approach can be extended as a general method for making conductive yarns and fabrics from all kinds of natural fibers.


Estimating efficacy in a randomized trial with product nonadherence: application of multiple methods to a trial of preexposure prophylaxis for HIV prevention.
Antiretroviral preexposure prophylaxis (PrEP) for persons at high risk of human immunodeficiency virus infection is a promising new prevention strategy. Six randomized trials of oral PrEP were recently conducted and demonstrated efficacy estimates ranging from 75% to no effect, with nonadherence likely resulting in attenuated estimates of the protective effect of PrEP. In 1 of these trials, the Partners PrEP Study (Kenya and Uganda, 2008-2011), participants (4,747 serodiscordant heterosexual couples) were randomized to receipt of tenofovir (TDF), coformulated TDF/emtricitabine (FTC), or placebo. Intention-to-treat analyses found efficacy estimates of 67% for TDF and 75% for TDF/FTC. We applied multiple methods to data from that trial to estimate the efficacy of PrEP with high adherence, including principal stratification and inverse-probability-of-censoring (IPC) weights. Results were further from the null when correcting for nonadherence: 1) among the strata with an estimated 100% probability of high adherence (TDF hazard ratio (HR) = 0.19, 95% confidence interval (CI): 0.07, 0.56; TDF/FTC HR = 0.12, 95% CI: 0.03, 0.52); 2) with IPC weights used to approximate a continuously adherent population (TDF HR = 0.18, 95% CI: 0.06, 0.53; TDF/FTC HR = 0.15, 95% CI: 0.04, 0.52); and 3) in per-protocol analysis (TDF HR = 0.18, 95% CI: 0.06, 0.53; TDF/FTC HR = 0.16, 95% CI: 0.05, 0.53). Our results suggest that the efficacy of PrEP with high adherence is over 80%.


The terminal arborization pattern of primary afferent fibers in the substantia gelatinosa of the spinal cord in the cat.
The spinal cords of adult cats were impregnated using the Golgi-Kopsch technique and cut in the sagittal plane. Richly arborizing axon-trees were found in the substantia gelatinosa (lamina II of Rexed). Axon arborizations, characterized by a long rostro-caudal course, a single-stem fiber, multiple-arborizing side branches and large terminal and en passant boutons, were subjected to a semi-quantitative and later to a computer-aided analysis. These arborizations, presumably of primary afferent origin (Réthelyi, 1977), are arranged in narrow sheets. The sheets of the individual arborizations are tilted with respect to the sagittal plane (the dorsal edge of the sheet having a more lateral position than the ventral edge), and tilted with respect to each other's plane, but they are rigorously oriented in the rostro-caudal direction. The average thickness of the sheets encompassing the terminal and en passant bulbs of these arborizations is 16.65 micrometer (S.D. = 2.86; n = 10). Occasionally, the side branches and terminal bulbs of a single stem fiber arborize into two separate sheets. These anatomical results suggest that the activity of the peripheral fibers spreads in long and narrow neuropil fields in the substantia gelatinosa. This pattern suggests a fine scale somatotopic arrangement.


Newborn circumcision.
Routine circumcision of male newborns continues to be practiced widely in the United States, although many physicians believe there is little medical indication for the procedure. The parents' decision regarding circumcision is often based on inadequate information; however, attempts to provide information to parents have not had much impact on the frequency of circumcision. Many third-party payers have begun to refuse payment for the procedure, and there are indications that this action will diminish the number of circumcisions.


Crystal structure and DFT study of 8-hy-droxy-1,2,3,5,6,7-hexa-hydro-pyrido[3,2,1-ij]quinoline-9-carbaldehyde.
In the title compound, C13H15NO2, the fused non-aromatic rings of the julolidine moiety adopt envelope conformations. The hy-droxy group forms an intra-molecular hydrogen bond to the aldehyde O atom, generating an S(6) ring motif. Weak inter-molecular C-H⋯O hydrogen bonds help to stabilize the crystal structure. Density functional theory (DFT) optimized structures at the B3LYP/6-311 G(d,p) level are compared with the experimentally determined mol-ecular structure in the solid state.


[Calcifications visible in mammographic examination as a diagnostic problem].
Mammography can detect various types of calcification in the breast. Their correct estimation enables detection of the breast cancer in situ. In our research there have been analyzed various types of calcification available on radiograms, which have been placed in publications. The aim was to find out which of them could be characteristic for cancer. Calcifications that are seen in mammography can be divided into: microcalcifications, macrocalcifications in galactophorous ducts, in blood and sympatic vessels. Pathogenesis of microcalcifications is not explained. There exist hypotheses which claim that they are caused by focuses of necrosis, changes in acid mucopolysaccharides in the midst of galactophorous ducts or in mitochondria. Differentiation between microcalcifications and macrocalcifications is based on variations in size, shape and radiological changes which they accompany.


Refractory very early-onset inflammatory bowel disease associated with cytosolic isoleucyl-tRNA synthetase deficiency: A case report.
Aminoacyl tRNA synthetases/ligases (ARSs) are highly conserved enzymes involved in attaching amino acids to tRNA promoting protein synthesis. Although deficiencies of ARSs localized to the mitochondria classically present with neuropathology, the clinical features of cytosolic ARS deficiencies are more variable. They have previously been associated with neonatal hepatitis, but never with early-onset inflammatory bowel disease. A nine-year-old Bangladeshi boy presented with neonatal liver failure and deranged clotting, transaminitis and cholestasis. His parents were first cousins. Two older brothers and a sister were well. The patient suffered from loose stools from early infancy which became more troublesome and persistent from five years old with ten bloody motions a day. Repeated endoscopies showed persistent pancolitis, which was refractory to mesalazine, corticosteroids, azathioprine, sirolimus and anti-TNF (adalimumab) therapy, but has improved recently with subcutaneous methotrexate.Whole Genome Sequencing revealed a novel pathogenic missense variant (c.290A > G) in the cytosolic isoleucyl-tRNA synthetase gene, leading to an amino acid substitution (p.Asp97Gly). Pathogenic variants in other genes associated with inflammatory bowel disease (IBD) (ADAM17, EGFR, FOXP3, IL10RA, IL10RB, IL21R, NCF4, STAT3) were excluded. Cytokine assays demonstrated markedly elevated IL-2, IL-5, IL-13, IL-9 and IL-10 by the patient's CD4+ T-cells, while IL-17A, IL-17F, IFNβ were lower, and TNFα not significantly different when compared to healthy controls. This case report provides evidence that recessive mutations in cytosolic isoleucyl-tRNA synthetase are a novel monogenic cause of IBD, which should be considered, particularly in infants and children with a history of neonatal hepatitis and very early-onset IBD poorly responsive to treatment.


Multiparametric and multinuclear magnetic resonance imaging of human breast cancer: current applications.
The exploration of novel imaging methods that have the potential to improve specificity for the identification of malignancy is still critically needed in breast imaging. Changes in physiologic alterations of soft tissue water associated with breast cancer can be visualized by magnetic resonance (MR) imaging. However, it is unlikely that a single MR parameter can characterize the complexity of breast tissue. Techniques such as multiparametric MR imaging, proton magnetic resonance spectroscopic (MRSI) imaging, and 23Na sodium MR imaging when used in combination provide a comprehensive data set with potentially more power to diagnose breast disease than any single measure alone. A combination of MR, MRSI, and 23Na sodium MR parameters may be examined in a single MR imaging examination, potentially resulting in improved specificity for radiologic evaluation of malignancy.


An overview on fluid resuscitation and resuscitation endpoints in burns: Past, present and future. Part 1 - historical background, resuscitation fluid and adjunctive treatment.
An improved understanding of burn shock pathophysiology and subsequent development of fluid resuscitation strategies has led to dramatic outcome improvements in burn care during the 20th century. While organ hypoperfusion caused by inadequate resuscitation has become rare in clinical practice, there is growing concern that increased morbidity and mortality related to over-resuscitation is occurring more frequently in burn care. In order to reduce complications related to this concept of "fluid creep", such as respiratory failure and compartment syndromes, efforts should be made to resuscitate with the least amount of fluid in order to provide adequate organ perfusion. In this first part of a concise review, historic and current evidence regarding the available fluids is discussed, as well as some adjunctive treatments modulating the inflammatory response. In the second part, special reference will be made to the role of abdominal hypertension in burn care and the endpoints used to guide fluid resuscitation will be discussed. Finally, as urine output has been recognized as a poor resuscitation target, a resuscitation protocol is suggested in part two which includes new targets and endpoints that can be obtained with modern, less invasive hemodynamic monitoring devices.


Comparison of swarming, mating performance and longevity of males Anopheles coluzzii between individuals fed with different natural fruit juices in laboratory and semi-field conditions.
It is assumed that malaria vectors feed on locally available nectar sources to obtain energy. Sugar feeding is energetically critical for the Anopheles male swarming and mating activities. However, little is known about the impact of local nectar feeding on male physiological development and its consequences on male mosquito life traits in the malaria control context. This study aimed to evaluate the influence of local fruit juices on the life traits of males Anopheles coluzzii. Swarming characteristics (number of males in swarm, number of mating pairs, and swarm duration) in semi-field conditions; mating rate and longevity in a laboratory setting were compared between males An. coluzzii fed exclusively with mango, papaya or banana juices. The trophic preference was investigated in semi-field conditions. The results of this study showed that in the laboratory, mosquitoes fed with papaya juices lived on average longer (10 days) than those fed with banana or mango juices (5 days) and had higher a mating rate (53%) than those fed with banana juice (40%). In the semi-field, the swarm size of mosquitoes fed with banana juice (85 males) was larger than that of mosquitoes fed with mango juice (60 males). The number of mating pairs formed from banana-fed male swarms (17 mating pairs) was higher than that formed from mango-fed male swarm (8 mating pairs). There was no difference in swarming duration between male treatments. Male mosquitoes had a preference for papaya and banana juices. The results indicate that the origin of plant-derived feeding is an important factor in the survival and reproduction of mosquitoes. This calls for further investigations of chemical contents of nectars and their impact on the physiological development of mosquitoes.


A heritable microduplication encompassing TBL1XR1 causes a genomic sister-disorder for the 3q26.32 microdeletion syndrome.
Recently, a new syndrome with intellectual disability (ID) and dysmorphic features due to deletions or point mutations within the TBL1XR1 gene located in the chromosomal band 3q26.32 has been described (MRD41, OMIM 616944). One recurrent point mutation in the TBL1XR1 gene has been identified as the cause of Pierpont syndrome (OMIM 602342), a distinct intellectual disability syndrome with plantar lipomatosis. In addition, different de novo point mutations in the TBL1XR1 gene have been found in patients with autism spectrum disorders (ASD) and intellectual disability. Here, we report four patients from two unrelated families in whom array-CGH analysis and real-time quantitative PCR of genomic DNA revealed a TBL1XR1-microduplication. Adjacent genes were not affected. The microduplication occurred as a de novo event in one patient, whereas the other three cases occurred in two generations of a second, unrelated family. We compare and contrast the clinical findings in TBL1XR1 microdeletion, point mutation, and microduplication cases and expand the TBL1XR1-associated phenotypic spectrum. ID, hearing loss, and ASD are common features of TBL1XR1-associated diseases. Our clinical observations add to the increasing evidence of the role of TBL1XR1 in brain development, and they simultaneously demonstrate that different genetic disease mechanisms affecting TBL1XR1 can lead to similar ID phenotypes. The TBL1XR1-microduplication syndrome is an intellectual disability/learning disability syndrome with associated incomplete penetrance ASD, hearing loss, and delay of puberty. Its phenotypic overlap indicates that it is a genomic sister-disorder to the 3q26.32 microdeletion syndrome.


Subcellular distribution of ependymins in goldfish brain measured by radioimmunoassay.
Goldfish CNS was fractionated by differential and density gradient centrifugation. The fractions obtained were characterized by marker enzymes typical of various subcellular organelles. They were further analyzed by radioimmunoassay for their contents of ependymins, two CNS glycoproteins known to participate in biochemical reactions after learning events. Ependymins were shown to be major constituents of the soluble cytoplasm (5.6% of the total protein content). The nuclear fraction was virtually devoid of ependymins (0.6% of protein). Small amounts were observed in the crude synaptosomal and microsomal fractions (1.0 and 3.5%, respectively). The highest steady-state concentration of ependymins, however, was measured in the brain extracellular fluid (15.6% of the protein), including the CSF. The specificity of the distribution was examined by intracerebroventricular injection of 125I-labeled ependymins as exogenous marker substances. No indication of an artificial redistribution of the radiolabel during homogenization and fractionation was obtained. The exogenous analogues of ependymins were, however, incorporated in vivo into organelles recovered in the nuclear and crude synaptosomal fractions. Our results suggest that ependymins may interact with synaptic membranes from the extracellular fluid, although so far no evidence for a specific receptor-type binding site could be obtained in vitro.


The role of erythropoietin in the anemia of myelodysplastic syndrome.
Although erythropoietin (EPO) deficiency is not responsible for the anemia of myelodysplasia, pharmacologic doses of recombinant human EPO (rHuEPO, epoetin alfa) and epoetin beta have been studied extensively as treatment of anemia in myelodysplastic syndrome (MDS). When an epoetin is used as a single growth factor in patients with MDS, clinically meaningful responses occur in only a small minority of patients (16%). Patients who are transfusion-dependent are less likely to respond than patients who are transfusion-independent. Serum EPO level has a weak association with response rate and cannot be used to select or exclude patients from empirical trials of epoetins. The dose schedule commonly used as initial treatment 40,000 U/week, is consistent with clinical observations, but an optimal dose schedule has not been determined. The combination of an epoetin and granulocyte colony-stimulating factor (G-CSF) produces a higher erythroid response rate (36%) than the regimen of epoetin alone, but we have found no randomized trial data to support this point. However, the design of the clinical trials, which included adding G-CSF after epoetin alone had failed, supports the hypothesis that combined use of growth factors, rather than just higher doses of epoetin, is responsible for the high response rate observed with the combination of epoetin and G-CSF. Unfortunately, as in the case of epoetin alone, patients who are transfusion-dependent (> or =2 U red blood cells/month) are less likely to respond to combined growth factor therapy. Although the ability of patients with MDS to show an erythroid response to epoetin is of biologic interest, because of high costs and the limited response rate in transfusion-dependent patients, epoetin therapy, with or without G-CSF, cannot be regarded as a definitive therapy for the anemia of MDS.


Occurrence of emerging mycotoxins in cereals and cereal-based products from the Korean market using LC-MS/MS.
The present study aimed to investigate the occurrence of emerging mycotoxins in cereals (n = 61) and cereal-based products (n = 36) collected from Korean market. First of all, using the quick, easy, cheap, effective, rugged and safe (QuEChERS) extraction method, and ultrahigh-pressure liquid chromatography (UPLC) with triple quadruple tandem mass spectrometry (MS/MS), we developed a simple and fast method for quantitative determination of eight emerging mycotoxins including alternariol (AOH), alternariol monomethyl ether (AME), tentoxin (TEN), beauvericin (BEA) and enniatins (ENs; ENA, ENA1, ENB and ENB1). The developed analytical method was validated in parameters of linearity, precision and accuracy. For UPLC-MS/MS analysis, the recoveries of emerging mycotoxins from spiked samples at three concentration levels ranged from 82.7% to 108.8% with RSDs between 0.4% and 14.7%. Analytical methods were applied to determine the contamination of mycotoxins in cereal and cereal-based product samples. Sixty-three of the total 97 samples were contaminated with at least one emerging mycotoxin. The maximum number of emerging mycotoxins observed in a single sample was six out of eight analytes. The highest level of contamination was detected in cereal at 70.9 μg/kg for alternariol monomethyl ether (AME). However, currently there is no international standard for emerging mycotoxins in food. Accordingly, it is necessary to establish a database of emerging mycotoxins contamination through continuous monitoring.


Peste des petits ruminants.
Peste des petits ruminants virus causes a highly infectious disease of small ruminants that is endemic across Africa, the Middle East and large regions of Asia. The virus is considered to be a major obstacle to the development of sustainable agriculture across the developing world and has recently been targeted by the World Organisation for Animal Health (OIE) and the Food and Agriculture Organisation (FAO) for eradication with the aim of global elimination of the disease by 2030. Fundamentally, the vaccines required to successfully achieve this goal are currently available, but the availability of novel vaccine preparations to also fulfill the requisite for differentiation between infected and vaccinated animals (DIVA) may reduce the time taken and the financial costs of serological surveillance in the later stages of any eradication campaign. Here, we overview what is currently known about the virus, with reference to its origin, updated global circulation, molecular evolution, diagnostic tools and vaccines currently available to combat the disease. Further, we comment on recent developments in our knowledge of various recombinant vaccines and on the potential for the development of novel multivalent vaccines for small ruminants.


Aptamer-Integrated Scaffolds for Biologically Functional DNA Origami Structures.
The manufacture of DNA origami nanostructures with highly ordered functional motifs is of great significance for biomedical applications. Here, we present a robust strategy to produce customized scaffolds with integrated aptamer sequences, which enables direct construction of functional DNA origami structures. As we demonstrated, aptamers of various numbers and types were efficiently and stably integrated in user-defined positions of the scaffolds. Specifically, two different thrombin aptamer sequences were simultaneously inserted into the M13mp18 phage genome. The assembled functional DNA origami structures from this aptamer-integrated scaffold exhibited increased binding efficiency to thrombin and displayed more than 10-fold stronger resistance to exonuclease degradation than that produced using the traditional staple extension method. Additionally, a scaffold integrated with the platelet-derived growth factor aptamer was produced, and the assembled DNA origami structures showed significant inhibitory effect on breast cancer cells MDA-MB-231. This scalable method of creating design-specific scaffolds opens up a new way to construct more stable and functionally robust DNA origami structures and thus provides an important basis for their broader applications.


Exploring the relationship between North Star Ambulatory Assessment and Health Utilities Index scores in Duchenne muscular dystrophy.
The North Star Ambulatory Assessment (NSAA) documents motor performance in ambulatory individuals with Duchenne muscular dystrophy (DMD). Health Utilities Index (HUI) scores, reflecting preferences for health-related quality-of-life (HRQoL) implications of health states, are commonly estimated within trials. This study sought to characterize the relationship between the NSAA score and utility in DMD. Family members serving as proxy respondents for placebo-treated ambulatory individuals with DMD (NCT01254019; BioMarin Pharmaceuticals Inc) completed the HUI and the NSAA (score range, 0-34). Mean change over time on these measures was estimated, and the correlation between changes in NSAA score and a) HUI utility; b) HUI3 ambulation and HUI2 mobility attribute scores, over 48 weeks was calculated. Baseline mean (range) age was 8.0 years (5-16; n = 61) and mean (standard deviation [SD]) scores were 0.87 (0.13; HUI2), 0.82 (0.19; HUI3), and 21.0 (8.1; NSAA). Mean (SD) change over 48 weeks was -0.05 (0.14; HUI2), -0.06 (0.19; HUI3), and -2.9 (4.7; NSAA). Weak positive correlations were observed between baseline NSAA score and HUI utility (HUI2: r = 0.29; HUI3: r = 0.17) and for change over 48 weeks (HUI2: r = 0.16; HUI3: r = 0.15). Stronger correlations were observed between change in NSAA score and the HUI3 ambulation (r = 0.41) and HUI2 mobility (r = 0.41) attributes. Among ambulatory individuals with DMD, NSAA score is weakly correlated with HUI utility, suggesting that motor performance alone does not fully explain HRQoL. Stronger relationships were observed between HUI ambulation and mobility attributes, and NSAA. Although unidimensional measures like the NSAA are informative for documenting disease-specific health impacts, they may not correlate well with measures of overall health status; requiring use in conjunction with other patient-reported and preference-based outcomes.


The evaluating self-management and educational support in severely obese patients awaiting multidisciplinary bariatric care (EVOLUTION) trial: principal results.
In Canada, demand for multidisciplinary bariatric (obesity) care far outstrips capacity. Consequently, prolonged wait times exist that contribute to substantial health impairments. A supportive, educational, self-management intervention (with in-person and web-based versions) for patients wait-listed for bariatric care has already been implemented in Northern and Central Alberta, Canada, but its effectiveness is unknown. The objective of this trial is to evaluate the clinical and economic outcomes of two self-management programs of varying intensity that are currently in use. We conducted a pragmatic, prospective, parallel-arm, randomized controlled trial of 651 wait-listed patients from two regional bariatric programs. Patients were randomized to (1) an in-person, group-based intervention (13 sessions; n = 215) or (2) a web-based intervention (13 modules; n = 225) or (3) control group (printed educational materials; n = 211). After randomization, subjects had 3 months to review the content assigned to them (the intervention period) prior to bariatric clinic entry. The primary outcome was the proportion of patients achieving 5% weight loss at 9 months. Intention-to-treat two-way comparisons were performed and adjusted for baseline age, sex, site and body mass index. At baseline, mean age was 40.4 ± 9.8 years, mean weight was 134.7 ± 25.2 kg, mean body mass index was 47.7 ± 7.0 kg/m2 and 83% of participants were female. A total of 463 patients (71%) completed 9 months follow-up. At least 5% weight loss was achieved by 24.2% of those in the in-person strategy, 24.9% for the web-based strategy and 21.3% for controls (adjusted p value = 0.26 for in-person vs. controls, 0.28 for web-based vs. controls, 0.96 for in-person vs. web-based). Absolute and relative (% of baseline) mean weight reductions were 3.7 ± 7.1 kg (2.7 ± 5.4%) for in-person strategy, 2.8 ± 6.7 kg (2.0 ± 4.8%) for web-based and 2.9 ± 8.8 kg (1.9 ± 5.9%) for controls (p > 0.05 for all comparisons). No between-group differences were apparent for any clinical or humanistic secondary outcomes. Total annual costs in Canadian dollars were estimated at $477,000.00 for the in-person strategy, $9456.78 for the web-based strategy and $2270.31 for provision of printed materials. Two different self-management interventions were no more effective and were more costly than providing printed education materials to severely obese patients. Our findings underscore the need to develop more potent interventions and the importance of comprehensively evaluating self-management strategies before widespread implementation. ClinicalTrials.gov, NCT01860131 . Registered 17 May 2013.


Aucuba japonica Extract and Aucubin Prevent Desiccating Stress-Induced Corneal Epithelial Cell Injury and Improve Tear Secretion in a Mouse Model of Dry Eye Disease.
Dry eye disease is affected by a broad range of causes such as age, lifestyle, environment, medication and autoimmune diseases. These causes induce tear instability that activates immune cells and promotes expression of inflammatory molecules. In this study, we investigated the therapeutic effects of an ethanolic extract of Aucuba japonica (AJE) and its bioactive compound, aucubin, on dry eye disease. The human corneal cells were exposed to desiccation stress induced by exposing cells to air, so that viability was decreased. On the other hand, pre-treatment of AJE and aucubin restored cell survival rate depending on the dose under the dry condition. This result was confirmed again by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining. The mRNA expression of inflammatory molecules was reduced by the pretreatment of AJE and aucubin under the dry state. The therapeutic effects of AJE and aucubin were examined in the animal model for dry eye induced by unilateral excision of the exorbital lacrimal gland. Declined tear volumes and corneal irregularity in the dry eye group were fully recovered by the administration of AJE and aucubin. The apoptotic cells on the cornea were also decreased by AJE and aucubin. Therefore, this study suggests that administration of AJE can be a novel therapeutic for dry eye disease and that the pharmacological activities of AJE may be in part due to its bioactive compound, aucubin.


Interanimal variability in preexisting collaterals is a major factor determining outcome in experimental angiogenesis trials.
Despite numerous animal trials reporting that cell therapy promotes collateral flow, clinical trials have not convincingly shown benefit. Patient-related risk factors are often used to explain these discrepancies. However, during the course of our own angiogenesis studies using mice, we noted large anatomical variability in collateral vessels. The purpose of the present investigation was to define how important this factor might be in determining intervention outcomes. Hindlimb ischemia was induced in BALB/c mice by ligating the superficial femoral artery. After 24 h, animals were treated by injecting the adductor muscle with either control media or cultured mesenchymal stem cells (MSCs). Blood flow recovery was measured using laser-Doppler [laser-Doppler perfusion imaging (LDPI) ratio]. In a second experiment, mice were stratified 24 h after arterial ligation before treatment by using a simple clinical score of the ligated leg: 1, able to flex, mild discoloration; 2, no flexion, mild discoloration; 3, severe discoloration; and 4, any necrosis. Without stratification, blood flow recovery significantly increased in the MSC-treated group (P < 0.05, n = 6 MSC group, n = 7 media group). In the experiment employing stratification, all differences between the groups disappeared (n = 11 MSC group, n = 10 media group; P = 0.3). Furthermore, we found a striking inverse correlation between clinical score on day 1 and the LDPI ratio on day 28 (P < 0.0001; n = 79). Anatomical confirmation of the disparity in preexisting collaterals was found in two different mouse strains using microscopic computed tomography. In conclusion, there is substantial interanimal variability in preexisting collateral flow, and this variability can importantly influence outcome. To overcome this, either animals must be stratified before treatment, the number of animals must be increased substantially, or, preferably, both.


Group contributions for estimating standard gibbs energies of formation of biochemical compounds in aqueous solution.
A method is presented for the estimation of the standard Gibbs energies of formation of biochemical compounds (and hence the Gibbs energies and equilibrium constants of biochemical reactions) from the contributions of groups. The method employs a large set of groups and special corrections. The contributions were estimated via multiple linear regression, using screened and weighted literature data. For most of the data employed, the error is less than 2 kcal/mol. The method provides a useful first approximation to Gibbs energies and equilibrium constants in biochemical systems.


Pan-cancer genetic analysis of cuproptosis and copper metabolism-related gene set.
A recent paper has revealed a novel cell death pathway, cuproptosis, a programmed cell death based on copper. This study aimed to evaluate the pan-cancer genomics and clinical association of cuproptosis and copper metabolism-related cell death genes, including SLC25A3, SLC25A37, SLC31A1, FDX1, DLAT, LIAS, ATP7A, ATP7B, COX17, SCO1, SCO2, COX11, and COX19. By mining multi-omics profiling data, we performed a comprehensive and systematic characterization of cuproptosis genes across more than 9,000 samples of over 30 types of cancer. ATP7B and ATP7A were the two most frequently mutated copper cell death genes in cancer. UCEC and SKCM were the two cancer types that have the highest mutation rates while the mutation of LIAS was associated with worse survival of BRCA. Brain cancer was potentially affected by copper cell death because of the difference in copper cell death gene expression among subtypes and stages. On the contrary, KIRC might have a lower cuproptosis activity because of the decrease in copper cell death gene expression. In lung cancer and kidney cancer, most of the cancer-noncancer expression patterns of copper cell death genes were consistent between mRNA and protein levels. Some of the cuproptosis gene expression was associated with the survival of LGG, KIRC, and ACC. The top five expression-copy numbers correlating cancer types were BRCA, OV, LUSC, HNSC, BLCA, and LUAD. Generally, the copy number variations of these genes in KIRC, UCEC, and LGG were associated with survival. The expression of DLAT, LIAS, and ATP7B was negatively correlated with the methylation in most of the cancer types. The copper cell death genes regulating miRNA and pathway regulation networks were constructed. The copper cell death genes were correlated with immune cell infiltration levels of multiple immune cells. These genes were correlated with the sensitivity of cancer cells to multiple drugs. Copper cell death genes are potentially involved in many cancer types and can be developed as candidates for cancer diagnosis, prognosis, and therapeutic biomarkers.


The natural history of untreated "prostatism".
One hundred and seven patients with symptoms of prostatic obstruction in whom prostatectomy was not clinically indicated were reviewed after an interval of 5 years. Ten had subsequently required surgery and 97 remained untreated. In the majority, symptoms did not worsen and only 2 developed acute retention. Urodynamic studies, where repeated, showed little change. Although there was no certain indicator or progression, a flow rate measurement could identify the patients at risk of clinical deterioration sufficient to require surgery.


A meta-analysis of case-control studies examining sporadic campylobacteriosis in Australia and New Zealand from 1990 to 2016.
We conducted a meta-analysis of case-control studies to identify locally relevant risk factors for sporadic campylobacteriosis in Australia and New Zealand. We searched Medline, Web of Science, ProQuest and Google Scholar using PRISMA guidelines. Reference lists and grey literature were hand-searched. Meta-analyses were conducted in the R package 'metafor' using published odds ratios and 95% confidence intervals. We identified 325 articles, from which we included 10 that described case-control studies. Four risk factors were statistically significant in the meta-analysis: eating undercooked poultry (OR=4.28, 95%CI 3.09-5.93); eating poultry cooked outside the home (OR=2.13, 95%CI 1.66-2.72); having pet chickens (OR=3.29, 95%CI 2.12-5.10); and overseas travel (OR=5.55, 95%CI 3.20-9.63). Among children, having pet dogs showed elevated but not significant risk (OR=1.57, 95%CI 0.99-2.49). We identified consumption of chicken meat and contact with domestic chickens as important risk factors for campylobacteriosis in Australia and New Zealand. Implications for public health: While consumption of chicken meat is a well-known risk factor for campylobacteriosis, zoonotic transmission is often overlooked. This research indicates a greater need for public health awareness surrounding zoonotic campylobacteriosis, especially for young children.


The Npc2Gt(LST105)BygNya mouse signifies pathological changes comparable to human Niemann-Pick type C2 disease.
Niemann-Pick type C2 disease (NP-C2) is a fatal neurovisceral disorder caused by defects in the lysosomal cholesterol transporter protein NPC2. Consequently, cholesterol and other lipids accumulate within the lysosomes, causing a heterogeneous spectrum of clinical manifestations. Murine models are essential for increasing the understanding of the complex pathology of NP-C2. This study, therefore, aims to describe the neurovisceral pathology in the NPC2-deficient mouse model to evaluate its correlation to human NP-C2. Npc2-/- mice holding the LST105 mutation were used in the present study (Npc2Gt(LST105)BygNya). Body and organ weight and histopathological evaluations were carried out in six and 12-week-old Npc2-/- mice, with a special emphasis on neuropathology. The Purkinje cell (PC) marker calbindin, the astrocytic marker GFAP, and the microglia marker IBA1 were included to assess PC degeneration and neuroinflammation, respectively. In addition, the pathology of the liver, lungs, and spleen was assessed using hematoxylin and eosin staining. Six weeks old pre-symptomatic Npc2-/- mice showed splenomegaly and obvious neuropathological changes, especially in the cerebellum, where initial PC loss and neuroinflammation were evident. The Npc2-/- mice developed neurological symptoms at eight weeks of age, severely progressing until the end-stage of the disease at 12 weeks. At the end-stage of the disease, Npc2-/- mice were characterized by growth retardation, tremor, cerebellar ataxia, splenomegaly, foam cell accumulation in the lungs, liver, and spleen, brain atrophy, pronounced PC degeneration, and severe neuroinflammation. The Npc2Gt(LST105)BygNya mouse model resembles the pathology seen in NP-C2 patients and denotes a valuable model for increasing the understanding of the complex disease manifestation and is relevant for testing the efficacies of new treatment strategies.


Prevalence, awareness, treatment, and control of hypertension among Ghanaian population in Amsterdam, The Netherlands: the GHAIA study.
Migration from sub-Saharan Africa to industrialized countries has increased tremendously over the last few decades, yet very little is known about the health status of sub-Saharan African populations living in industrialized communities. The aim of this study was to assess prevalence, levels of awareness, treatment, and control of hypertension among the largest sub-Saharan African group (Ghanaians) living in the Netherlands. Cross-sectional study of Ghanaian adults aged 18-60 years in Amsterdam, the Netherlands. The overall prevalence of hypertension was 55%. Of these, about half were aware of their condition, 45% were receiving antihypertensive medication, and 15% were controlled (blood pressure <140/90 mmHg). The prevalence rates of hypertension, awareness, and treatment were similar among males and females. However, males had a lower blood pressure control rate than females. Among all hypertensives, 22% of females had their blood pressure controlled compared with 5.8% in males: adjusted prevalence ratio (APR) 3.94 (95% CI 1.05-14.79). Among those receiving treatment for their hypertension, 48% of females were controlled compared with only 13% of males: APR 4.08 (95% CI 1.20-13.87). Hypertension is a major problem among this recently migrated sub-Saharan African population. Furthermore, hypertension control rate is very low particularly in males. Urgent measures are needed to halt the increasing prevalence of hypertension and to improve hypertension control among these populations.


Management of the COVID-19 Pandemic in the Republic of Korea from the Perspective of Governance and Public-Private Partnership.
The government of the Republic of Korea (ROK) has established relevant laws and a master plan for infectious disease control and prevention based on their experiences with past epidemics. In addition, the Ministry of Health and Welfare has designed a standard manual for risk management that involves pan-governmental crisis management systems. This national infectious disease management system is constantly being supplemented and developed in face of infectious disease-related crises. In this study, we set out to present directions for infectious disease prevention and flow of management and governance between central and local governments to ensure systematic quarantine activities in the ROK. During the coronavirus disease 2019 (COVID-19) pandemic, public-private partnerships have been established to collect, provide, process, and disseminate information for effective quarantine. This has enabled the development and rapid approval of test kits, the tracking of cases, and the allocation of appropriate resources for patient treatment. Additionally, the Infectious Disease Control Agency has actively utilized information and communication technology platforms to disclose information necessary in real-time for COVID-19 quarantine and management. Overall, these efforts have played an important role in epidemiological investigations, patient management, and quarantine in the early stages of the COVID-19 pandemic.


Herpes simplex virus type 1 ICP-0 induces reactivation of pseudorabies virus from latently infected trigeminal ganglia explant cultures.
Pseudorabies virus (PrV), like other alphaherpesviruses, is a neurotropic virus that can establish a latent infection in swine. Reactivation of PrV from latency may occur spontaneously or after induction with corticosteroids. The mechanisms involved in the establishment of latency and reactivation are currently unknown. Here, we examined gene-specific reactivation of PrV by herpes simplex virus type 1 (HSV-1) immediate early protein, ICP-0. Primary neuronal cell cultures established from the trigeminal ganglia of latently infected swine were superinfected with recombinant adenoviruses expressing ICP-0. Reactivation of PrV occurred in cultures that were superinfected with two different ICP-0-expressing adenovirus recombinants, but not in cultures that were either mock-infected, or superinfected with wild-type adenovirus, or recombinant adenoviruses not expressing ICP-0. Infectious PrV was detected between 4 and 7 days postinfection, regardless of the promoter driving expression of ICP-0. Results from these experiments show that HSV-1 ICP-0, a homologue of PrV EP0, can induce PrV reactivation from explanted trigeminal ganglia of latently infected swine.


Japanese herbal medicine, Saiko-keishi-to, prevents gut ischemia/reperfusion-induced liver injury in rats via nitric oxide.
To determine whether Saiko-keishi-to (TJ-10), a Japanese herbal medicine, could protect liver injury induced by gut ischemia/reperfusion (I/R), and to investigate the role of NO. Male Wistar rats were exposed to 30-min gut ischemia followed by 60 min of reperfusion. Intravital microscopy was used to monitor leukocyte recruitment. Plasma tumor necrosis factor (TNF) levels and alanine aminotransferase (ALT) activities were measured. TJ-10 1 g/(kg.d) was intragastrically administered to rats for 7 d. A NO synthase inhibitor was administered. In control rats, gut I/R elicited increases in the number of stationary leukocytes, and plasma TNF levels and ALT activities were mitigated by pretreatment with TJ-10. Pretreatment with the NO synthase inhibitor diminished the protective effects of TJ-10 on leukostasis in the liver, and the increase of plasma TNF levels and ALT activities. Pretreatment with TJ-10 increased plasma nitrite/nitrate levels. TJ-10 attenuates the gut I/R-induced hepatic microvascular dysfunction and sequential hepatocellular injury via enhancement of NO production.


Neoantigens in complement component C3 as detected by monoclonal antibodies. Mapping of the recognized epitopes by synthetic peptides.
The different fragments of the third complement component, C3, generated upon complement activation/inactivation have the ability to bind to several other complement components and receptors as well as to proteins of foreign origin. These multiple reactivities of C3 fragments are associated with a series of conformational changes occurring in the C3 molecule during its degradation. The conformations acquired by the different C3 fragments are also associated with the exposure of neoantigenic epitopes that are specific for (a) particular fragment(s). In order to study these epitopes and thus the conformational changes occurring in C3, monoclonal antibodies (mAbs) recognizing such epitopes were produced in Balb/c mice after immunization with denatured human C3. Two of the three antibodies (7D84.1 and 7D264.6) presented in this study recognized predominantly surface-bound iC3b, and one mAb (7D323.1) recognized both surface-bound and fluid-phase iC3b. Although none of the mAbs recognized any other fluid-phase C3 fragment, all three antibodies detected micro-titre-plate-fixed C3b and iC3b, but not C3c or C3d. In addition to the reaction with human C3, mAb 7D323.1 also bound to micro-titre-plate-fixed rabbit C3. The epitopes recognized by the three mAbs were further localized by using synthetic peptides that were designed on the basis of the differential binding of the mAbs to the C3 fragments. All three antibodies reacted with C3-(924-965)-peptide, which represents the region of C3 between the kallikrein-cleavage site (923-924) and the elastase-cleavage site (965-966). On the basis of the binding of the mAbs to five different overlapping peptides spanning the region between residues 924 and 965 of the human C3 sequence, and the sequence similarity between human C3 and rabbit C3 within this area, the epitopes recognized by these antibodies are mapped. The contribution of the individual amino acid residues in the formation of the epitopes is discussed.


Three-Component Coupling of Anilines, Amines, and Difluorocarbene to Access Formamidines.
A one-pot three-component reaction of two anilines (or one aniline and one alkylamine) and in situ-generated difluorocarbene is developed herein to enable efficient construction of formamidines. Crucial formimidoyl fluoride intermediate RN═CHF is proposed from the reaction of a primary aniline and difluorocarbene. Ensuing nucleophilic iminyl substitution of this intermediate with a second amine allows cross-condensation of the two amines to produce formamidines. When only one type of primary aniline is used as the substrate, the difluoromethylated homo-condensation products can also be produced under a 1:1 molar ratio of aniline/difluorocarbene. Intramolecular variant of this method allows concise synthesis of benzimidazoquinazolines and nitrogen-fused/spirocyclic compounds, showing the potential of this method in organic synthesis. More interesting reactions are anticipated by exploiting the reactivity of difluorocarbene and primary amines to isocyanides or the formimidoyl fluoride intermediates.


Speckle-based optical encryption with complex-amplitude coding and deep learning.
We propose a speckle-based optical encryption scheme by using complex-amplitude coding and deep learning, which enables the encryption and decryption of complex-amplitude plaintext containing both amplitude and phase images. During encryption, the amplitude and phase images are modulated using a superpixel-based coding technique and feded into a digital micromirror device. After passing through a 4f system, the information undergoes disturbance modulation by a scattering medium, resulting in a diffracted speckle pattern serving as the ciphertext. A Y-shaped convolutional network (Y-Net) model is constructed to establish the mapping relationship between the complex-amplitude plaintext and ciphertext through training. During decryption, the Y-Net model is utilized to quickly extract high-quality amplitude and phase images from the ciphertext. Experimental results verify the feasibility and effectiveness of our proposed method, demonstrating that the potential of integrating speckle encryption and deep learning for optical complex-amplitude encryption.


The Solute Polarization and Structural Effects on the Nonlinear Optical Response of Based Chromone Molecules.
The solute polarization due to solvent is a an electrostatic quantum effect that impacts diverse molecular properties, including the nonlinear optical response of a material. An iterative procedure that allows updating the solute charge distribution in the presence of the solvent is combined with a sequential Monte Carlo/Quantum Mechanics methodology and Density Functional Theory methods to evaluate the nonlinear optical (NLO) response using the hyper Rayleigh scattering (HRS) of a series of chromones recently identified in Chamaecrista diphylla, an herbaceous plant abundant throughout the Americas and used in folk medicine. From this study, it is determined that from gas to solvent environment, the systems acquire low refractive index (n) and an improvement of the first hyperpolarizability (βHRS ), signaling potential NLO uses. It is shown that the octupolar contributions (βJ=3 ) superate the dipolar ones (βJ=1 ) and dominate the second-order optical response in both gas and liquid phases, which indicate nontrivial optical materials. Moreover, the solvent environment and structural changes in the periphery can tune significantly the dipolar/octupolar balance, showing a key to control the decoupling between these contributions.


Why having a voice is important to children who are involved in family support services.
Having a voice in family matters is considered a protective factor from harm, and key to promoting children's wellbeing. However, since the adoption of the United Nations Convention on the Rights of the Child (1989) and specifically Article 12 pertaining to children's participation, research reveals that children's voices often remain invisible in child protection and family welfare services. While there is renewed interest in hearing children's voices about their experiences in out-of-home care, there remains little awareness and knowledge of children's voices in family support services. This article addresses this gap by presenting children's own meanings and experiences of having a voice, derived from a research collaboration between UnitingCare and Queensland University of Technology . 17 children aged 6-16 years (8 sibling groups) whose families received family support services (voluntary or ordered) from UnitingCare in Queensland, Australia. An interpretative phenomenological approach with activity-based interviews involving art, play and Reflexions cards were used to support children to share their lived experiences. Children's sense-making related to having a voice revealed four connected meanings that poignantly illustrated that not only should children have a say about the supports they receive because they are part of a family, but because they are knowledgeable agents with insights that can improve their and their family's experiences. Family support services need a sustained paradigm shift towards protection with participation to incorporate the voices of children as an everyday practice to ensure their wellbeing and safety.


Noncontiguous finished genome sequences and description of Bacillus massiliglaciei, Bacillus mediterraneensis, Bacillus massilinigeriensis, Bacillus phocaeensis and Bacillus tuaregi, five new species identified by culturomics.
Microbial culturomics, which investigates microbial diversity by combining diversified culture conditions, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and 16S rDNA identification, allowed to identify five new species within the Bacillus genus. Bacillus massiliglaciei strain Marseille-P2600T, Bacillus mediterraneensis strain Marseille-P2384T, Bacillus massilinigeriensis strain Marseille-P2366T, Bacillus tuaregi strain Marseille-P2489T and Bacillus phocaeensis strain SIT16T are each the type strain of the corresponding bacterial species. These strains, the genomes of which are described here, are facultative anaerobic Gram-positive bacilli. Here, we describe the main characteristics of each bacterium and present their complete genome sequence and annotation.


An In Vitro Investigation of Platelet-Rich Plasma-Gel as a Cell and Growth Factor Delivery Vehicle for Tissue Engineering.
Platelet-rich plasma (PRP) has been used for different applications in human and veterinary medicine. Many studies have shown promising therapeutic effects of PRP; however, there are still many controversies regarding its composition, properties, and clinical efficacy. The aim of this study was to evaluate the influence of different platelet concentrations on the rheological properties and growth factor (GF) release profile of PRP-gels. In addition, the viability of incorporated bone marrow-derived human mesenchymal stem cells (MSCs) was investigated. PRP (containing 1000 × 10(3), 2000 × 10(3), and 10,000 × 10(3) platelets/μL) was prepared from human platelet concentrates. Platelet activation and gelification were achieved by addition of human thrombin. Viscoelastic properties of PRP-gels were evaluated by rheological studies. The release of GFs and inflammatory proteins was measured using a membrane-based protein array and enzyme-linked immunosorbent assay. MSC viability and proliferation in PRP-gels were assessed over 7 days by cell viability staining. Cell proliferation was examined using DNA quantification. Regardless of the platelet content, all tested PRP-gels showed effective cross-linking. A positive correlation between protein release and the platelet concentration was observed at all time points. Among the detected proteins, the chemokine CCL5 was the most abundant. The greatest release appeared within the first 4 h after gelification. MSCs could be successfully cultured in PRP-gels over 7 days, with the highest cell viability and DNA content found in PRP-gels with 1000 × 10(3) platelets/μL. The results of this study suggest that PRP-gels represent a suitable carrier for both cell and GF delivery for tissue engineering. Notably, a platelet concentration of 1000 × 10(3) platelets/μL appeared to provide the most favorable environment for MSCs. Thus, the platelet concentration is an important consideration for the clinical application of PRP-gels.


Sensitization of Escherichia coli B/r to X-irradiation by 2'-chloro-2'-deoxythymidine.
The sensitizing effect of 2'Cl-TdR has been investigated from the viewpoint of the formation of free radicals at the C-2' position of 2'Cl-TdR when reacted with hydrated electrons. E. coli B/r cells were incubated in growth medium containing 2'Cl-TdR. Centrifugation experiments using CsCl equilibrium density gradients were carried out to confirm the incorporation of 2'Cl-TdR into DNA of the cells. The sedimentation profiles of DNA from cells grown in a medium containing 2'Cl-TdR were found at relatively heavier density positions in comparison with those of DNA from control cells. This result confirms that 2'Cl-TdR was incorporated into DNA. E. coli B/r cells which incorporated 2'Cl-TdR were more sensitive than the control cells for killing by X-irradiation under aerobic conditions. On the other hand, no difference in U.V.-inactivation curves between the cells grown in growth medium containing 2'Cl-TdR and the control cells was observed. From experiments using alkaline sucrose density gradient centrifugation, an increase in the frequency of radiation-induced single strand breaks of E. coli DNA containing 2'Cl-TdR was observed, compared with those of DNA from control cells. These results reveal that the sensitization of 2'Cl-TdR originates from an increase of damage at the sugar moiety in DNA.


Draft Genome Sequence of Geobacter sp. Strain SVR, Isolated from Antimony Mine Soil.
We report here the draft genome sequence of Geobacter sp. strain SVR, isolated from antimony mine soil in Hyogo Prefecture, Japan. The genome sequence data in this study will provide useful information for understanding bacterial antimonate reduction.


Hb Chapel Hill or Alpha2 74(EF3) Asp>Gly, a mildly unstable variant found in a Chinese family.
Hb Chapel Hill [Alpha2 74(EF3) Asp > Gly] results from an GAC > GGC substitution at codon 74 of the HBA1 or HBA2 genes. Hb Chapel Hill has not been reported since 1986. A heterozygous mutation, HBA2: c.224A > G, was identified in the proband, her father and sister. We compared the haematological and clinical data of this family with the data reported in the limited number of individuals. Having excluded iron deficiency, the Hb Chapel Hill was asymptomatic in heterozygous state. The cases presented here characterize cases in new techniques including capillary electrophoresis (CE). Two aberrant peaks were identified by CE, a major peak migrating in the zone 7 that correspond to Hb Chapel Hill (αChapel Hill 2β2) and a minor peak migrating in the zone 1 that correspond to Hb Chapel Hill2 (αChapel Hill 2δ2). Focusing on the variant expression, the Hb Chapel Hill plus Hb A2 variant were around 18.9-20.6% of total Hb in three members. This data will be useful for providing up-to-date and high quality information on the Hb Chapel Hill.


Mass Spectrometry-Based Detection of Beta Lactam Hydrolysis Enables Rapid Detection of Beta Lactamase Mediated Antibiotic Resistance.
Antibiotic resistance by beta lactamase expression is a serious and growing threat. We aimed to determine whether beta-lactamase activity is detectable in urine specimens to enable faster identification of resistance. Urine specimens from patients with extended spectrum beta lactamase (ESBL)-expressing urinary infections were incubated with beta lactam antibiotics. Beta lactam hydrolysis was determined by mass spectrometry methods. Ceftriaxone hydrolysis was observed in 45 of 45 ESBL-containing specimens from patients not treated with a beta lactamase inhibitor before specimen collection. Ceftriaxone hydrolysis was not observed in 108 of 108 non-ESBL-containing specimens. Spiking studies show that beta lactam hydrolysis can be observed within 30 minutes. Beta lactam hydrolysis is evidenced by mass spectrometry preceded by either liquid chromatography or matrix-assisted laser desorption ionization specimen processing methods. Clinically significant beta lactamase activity is detectable directly from urine specimens. The described methods would enable the detection of beta lactam resistance 24 to 48 hours sooner than culture based methods.


Lack of human cytomegalovirus expression in single cells from glioblastoma tumors and cell lines.
The relationship between human cytomegalovirus (HCMV) and glioblastoma (GBM) is an ongoing debate with extensive evidence supporting or refuting its existence through molecular assays, pre-clinical studies, and clinical trials. We focus primarily on the crux of the debate, detection of HCMV in GBM samples using molecular assays. We propose that these differences in detection could be affected by cellular heterogeneity. To take this into account, we align the single-cell RNA sequencing (scRNA-seq) reads from five GBM tumors and two cell lines to HCMV and analyze the alignments for evidence of (i) complete viral transcripts and (ii) low-abundance viral reads. We found that neither tumor nor cell line samples showed conclusive evidence of full HCMV viral transcripts. We also identified low-abundance reads aligned across all tumors, with two tumors having higher alignment rates than the rest of the tumor samples. This work is meant to rigorously test for HCMV RNA expression at a single cell level in GBM samples and examine the possible utility of single cell data in tumor virology.


Long-term follow-up of patients undergoing postconditioning during ST-elevation myocardial infarction.
Reperfusion injury may offset the optimal salvage of myocardium achieved during primary coronary angioplasty. Thus, coronary reperfusion must be combined with cardioprotective adjunctive therapies in order to optimize myocardial salvage and minimize infarct size. Forty-three patients with their first ST-elevation myocardial infarction were randomized to myocardial postconditioning or standard of care at the time of primary coronary angioplasty. Postconditioning was performed immediately upon crossing the lesion with the guide wire and consisted of four cycles of 30 s occlusion followed by 30 s of reperfusion. End-points included infarct size, myocardial perfusion grade (MPG), left-ventricular ejection fraction (LVEF), and long-term clinical events (death and heart failure). Despite similar ischemic times (≅4.5 h) (p = 0.9) a reduction in infarct size was observed among patients treated with the postconditioning protocol. Peak creatine phosphokinase (CPK), as well as its myocardial band (MB) fraction, was significantly lower in the postconditioning group when compared with the control group (CPK--control, 2,444 ± 1,928 IU/L vs. PC, 2,182 ± 1,717 IU/L; CPK-MB--control, 242 ± 40 IU/L vs. PC, 195 ± 33 IU/L; p = 0.64 and p < 0.01, respectively). EF in the postconditioning group was improved when compared with the control group (control, 43% ± 15 vs. PC, 52% ± 9; p = 0.05). After a mean follow-up of 3.4 years, a 6-point absolute difference in LVEF was still evident in the postconditioning group (p = 0.18). MPG was better among patients treated with the postconditioning protocol compared with control (2.5 ± 0.5 vs. 2.1 ± 0.6; p = 0.02). Due to the small sample size no significant differences in clinical events were detected (p value for death = 0.9; p value for heart failure = 0.2). A simple postconditioning protocol applied at the onset of mechanical reperfusion, resulted in reduction of infarct size, better epicardial and myocardial flow, and improvement in left ventricular function. The beneficial effects of postconditioning on cardiac function persist beyond 3 years.


Effects of Supplemental Oxygen on Cardiovascular and Respiratory Interactions by Extended Partial Directed Coherence in Idiopathic Pulmonary Fibrosis.
Idiopathic pulmonary fibrosis (IPF) is a chronic and restrictive disease characterized by fibrosis and inflammatory changes in lung tissue producing a reduction in diffusion capacity and leading to exertional chronic arterial hypoxemia and dyspnea. Furthermore, clinically, supplemental oxygen (SupplO2) has been prescribed to IPF patients to improve symptoms. However, the evidence about the benefits or disadvantages of oxygen supplementation is not conclusive. In addition, the impact of SupplO2 on the autonomic nervous system (ANS) regulation in respiratory diseases needs to be evaluated. In this study the interactions between cardiovascular and respiratory systems in IPF patients, during ambient air (AA) and SupplO2 breathing, are compared to those from a matched healthy group. Interactions were estimated by time series of successive beat-to-beat intervals (BBI), respiratory amplitude (RESP) at BBI onset, arterial systolic (SYS) and diastolic (DIA) blood pressures. The paper explores the Granger causality (GC) between systems in the frequency domain by the extended partial directed coherence (ePDC), considering instantaneous effects. Also, traditional linear and nonlinear markers as power in low (LF) and high frequency (HF) bands, symbolic dynamic indices as well as arterial baroreflex, were calculated. The results showed that for IPF during AA phase: 1) mean BBI and power of BBI-HF band, as well as mean respiratory frequency were significantly lower (p < 0.05) and higher (p < 0.001), respectively, indicating a strong sympathetic influence, and 2) the RESP → SYS interaction was characterized by Mayer waves and diminished RESP → BBI, i.e., decreased respiratory sinus arrhythmia. In contrast, during short-term SupplO2 phase: 1) oxygen might produce a negative influence on the systolic blood pressure variability, 2) the arterial baroreflex reduced significantly (p < 0.01) and 3) reduction of RSA reflected by RESP → BBI with simultaneous increase of Traube-Hering waves in RESP → SYS (p < 0.001), reflected increased sympathetic modulation to the vessels. The results gathered in this study may be helpful in the management of the administration of SupplO2.


In vitro screening test using Leishmania promastigotes stably expressing mCherry protein.
Transgenic Leishmania major and Leishmania donovani axenic promastigotes constitutively expressing mCherry were used for in vitro antileishmanial drug screening. This method requires minimal sample manipulation and can be easily adapted to automatic drug tests, allowing primary high-throughput screenings without the need for expensive and sophisticated instruments.


Nanodelivery Systems as a Novel Strategy to Overcome Treatment Failure of Cancer.
Despite the tremendous progress in cancer treatment in recent decades, cancers often become resistant due to multiple mechanisms, such as intrinsic or acquired multidrug resistance, which leads to unsatisfactory treatment effects or accompanying metastasis and recurrence, ultimately to treatment failure. With a deeper understanding of the molecular mechanisms of tumors, researchers have realized that treatment designs targeting tumor resistance mechanisms would be a promising strategy to break the therapeutic deadlock. Nanodelivery systems have excellent physicochemical properties, including highly efficient tissue-specific delivery, substantial specific surface area, and controllable surface chemistry, which endow nanodelivery systems with capabilities such as precise targeting, deep penetration, responsive drug release, multidrug codelivery, and multimodal synergy, which are currently widely used in biomedical researches and bring a new dawn for overcoming cancer resistance. Based on the mechanisms of tumor therapeutic resistance, this review summarizes the research progress of nanodelivery systems for overcoming tumor resistance to improve therapeutic efficacy in recent years and offers prospects and challenges of the application of nanodelivery systems for overcoming cancer resistance.


Empirical macroscopic features of spatial-temporal traffic patterns at highway bottlenecks.
Results of an empirical study of congested patterns measured during 1995-2001 at German highways are presented. Based on this study, various types of congested patterns at on and off ramps have been identified, their macroscopic spatial-temporal features have been derived, and an evolution of those patterns and transformations between different types of the patterns over time has been found out. It has been found that at an isolated bottleneck (a bottleneck that is far enough from other effective bottlenecks) either the general pattern (GP) or the synchronized flow pattern (SP) can be formed. In GP, synchronized flow occurs and wide moving jams spontaneously emerge in that synchronized flow. In SP, no wide moving jams emerge, i.e., SP consists of synchronized flow only. An evolution of GP into SP when the flow rate to the on ramp decreases has been found and investigated. Spatial-temporal features of complex patterns that occur if two or more effective bottlenecks exist on a highway have been found out. In particular, the expanded pattern where synchronized flow covers two or more effective bottlenecks can be formed. It has been found that the spatial-temporal structure of congested patterns possesses predictable, i.e., characteristic, unique, and reproducible features, for example, the most probable types of patterns that are formed at a given bottleneck. According to the empirical investigations the cases of the weak and the strong congestion should be distinguished. In contrast to the weak congestion, the strong congestion possesses the following characteristic features: (i) the flow rate in synchronized flow is self-maintaining near a limit flow rate; (ii) the mean width of the region of synchronized flow in GP does not depend on traffic demand; (iii) there is a correlation between the parameters of synchronized flow and wide moving jams: the higher the flow rate out from a wide moving jam is, the higher is the limit flow rate in the synchronized flow. The strong congestion often occurs in GP whereas the weak congestion is usual for SP. The weak congestion is often observed at off ramps whereas the strong congestion much more often occurs at on ramps. Under the weak congestion diverse transformations between different congested patterns can occur.


An overview on Leucosidea sericea Eckl. & Zeyh.: A multi-purpose tree with potential as a phytomedicine.
Leucosidea sericea (the sole species in this genus) is a tree species found in southern Africa and possesses several therapeutical effects against infectious diseases in humans and livestock. This review aims to document and summarize the botany, phytochemical and biological properties of Leucosidea sericea. Using the term 'Leucosidea sericea', we systematically searched literature including library catalogues, academic dissertations and databases such as PubMed, SciFinder, Web of Science, Google Scholar and Wanfang. Taxonomy of the species was validated using 'The Plant List' (www.theplantlist.org). Leucosidea sericea remains a widely used species among the different ethnic groups in southern Africa. The species is a rich source of approximately 50 essential oils and different classes of phytochemicals (phenolics, phloroglucinols, cholestane triterpenoids, alkaloids and saponins) which may account for their diverse biological properties. Documented biological activities which were mainly observed under in vitro systems included antimicrobial, anti-parasitic, antioxidant, acetylcholinesterase (AChE) inhibition and anti-inflammatory properties. Preliminary safety tests on Leucosidea sericea extracts suggest moderate cytotoxic effects based on a few cell lines that were investigated. Leucosidea sericea possesses diverse medicinal potential with the antimicrobial, anti-parasitic, antioxidant and anti-inflammatory activities being the most prominent. The relative abundance and tendency of Leucosidea sericea to evade in nature suggest an abundant reservoir of raw materials for potential commercialization (upon validation of its pharmacological potential). However, more stringent investigations on the extracts (and isolated bioactive compounds) focusing on the mode of actions, which will inevitably unravel their pharmacokinetics, pharmacodynamics and clinical relevance remain pertinent.


In-Vitro Assessment of Magnetic Dextran-Spermine Nanoparticles for Capecitabine Delivery to Cancerous Cells.
Cationic polymeric nanoparticles have great potential for developing drug delivery systems with limited side effects for tumor medication. The goal of this research is investigation of cationic dextran-spermine polymer (DS) efficacy for improvement of hydrophilic drug delivery to negatively charged cancerous cells. Capecitabine (as a hydrophilic antineoplastic drug) was loaded into the magnetic dextran-spermine nanoparticles (DS-NPs) via ionic gelation. Design of experiments was applied to specify how the significant factors affect size, surface charge and capecitabine entrapment efficiency of the DS-NPs. Physicochemical properties, in-vitro release profile and cellular studies of the optimized DS-NPs were evaluated. The experimental results indicated that DS-NPs with favorable properties can be achieved at an optimized condition of 2 mg/mL DS and 0.75 mg/mL tri-polyphosphate (TPP) concentrations, TPP addition rate of 35 mL/min, pH 3 of DS solution and super paramagnetic iron oxide nanoparticles (SPION)/DS mass ratio of 0.5. The entrapment efficiency of capecitabine was 26.1% at optimum condition and drug release at neutral pH after 24 h and acidic pH within 3 h was 56 and 98%, respectively. The cytotoxicity assessment exhibited that capecitabine loaded DS-NPs was more toxic than corresponding free drug as control. Significant cellular uptake of capecitabine loaded DS-NPs by U87MG glioblastoma cells were proved by Prussian blue staining and TEM, qualitatively. DS-NPs are suitable candidates for delivery of the hydrophilic drugs in cancer treatment and due to positive charge of the dextran-spermine, the uptake of the hydrophilic drugs by the cancerous cells was improved.


Targeted chemotherapy with cytotoxic bombesin analogue AN-215 can overcome chemoresistance in experimental renal cell carcinomas.
Multidrug resistance (MDR) mediated by membrane transporters, such as P-glycoprotein (MDR-1) and MDR-associated protein (MRP), remains a challenge in the therapy of renal cell carcinoma (RCC). Chemotherapy targeted to hormone receptors may provide a new approach to overcome chemoresistance. The cytotoxic analogue of bombesin/gastrin-releasing peptide (GRP), AN-215, consists of a superactive derivative of doxorubicin, AN-201, which is linked to a bombesin analogue carrier: RC-3094. The authors examined the expression of bombesin/GRP receptors in 3 human RCC cell lines (A-498, ACHN. and 786-0) by using reverse-transcriptase-polymerase chain reaction (RT-PCR) analysis and radioligand-binding assays. They also evaluated the effects of AN-215 and its cytotoxic radical AN-201 in the same RCC models in vivo, and they studied the effects of AN-215 and AN-201 on the expression levels of MDR-1 and subtype 1 of MRP (MRP-1) by using real-time PCR. A N-215 significantly (P < 0.05) inhibited the growth of A-498, ACHN, and 786-0 RCC xenografted into nude mice by 59.2-67.6%, whereas the cytotoxic radical AN-201 alone had no significant antitumor effects. The efficacy of AN-215 was independent of the expression patterns of MDR-1 and MRP-1 in these RCC cell lines. The induction of MDR-1 by AN-215 was similar (Experiment 2) or weaker (Experiment 1) compared with AN-201. Both AN-215 and AN-201 caused only a minor induction of MRP-1. The current findings indicated that targeted chemotherapy with cytotoxic bombesin/GRP analogue AN-215 can inhibit the growth of RCC, providing a new treatment modality for patients with advanced RCC.


The combination of platelet count and lymphocyte to monocyte ratio is a prognostic factor in patients with resected breast cancer.
Many inflammation indicators have been reported to be related with patient outcomes in various cancers. Previous studies have evaluated the combination of platelet (PLT) and lymphocyte to monocyte ratio (COP-LMR) as a systemic inflammatory marker for prognostication in lung cancer, yet its prognostic role among breast cancer patients remains unclear.In the present study, a total of 409 breast cancer patients with surgical resection were retrospectively investigated. The receiver operating characteristic (ROC) curve was used to choose the optimal cut-off value of PLT and lymphocyte to monocyte ratio (LMR). Patients were classified into 3 groups according to the score of COP-LMR, and its relationship with various clinicopathological factors and breast cancer prognosis were further evaluated.The ROC curve analysis showed that COP-LMR had a higher area under the ROC curve for the prediction of 5-year disease-free survival and overall survival than PLT or LMR alone. Multivariable analysis showed that an elevated COP-LMR was an independent predictor of poor disease-free survival (P = .032) and overall survival (P = .005). Subgroup analysis revealed that COP-LMR was still significantly associated with prognosis in both luminal A and luminal B subtypes.Preoperative COP-LMR is a potential prognostic factor in breast cancer patients who underwent surgery.


The Role of Wnt and R-spondin in the Stomach During Health and Disease.
The Wnt signaling pathway is one of the most prominent developmental signals. In addition to its functions in development, there is emerging evidence that it is also crucial for various organ functions in adult organisms, where Wnt signaling controls tissue stem cell behavior, proliferation and differentiation. Deregulation of Wnt signaling is involved in various pathological conditions and has been linked to malignant tissue transformation in different organ systems. The study of the Wnt signaling pathway has revealed a complex regulatory network that tightly balances the quality and strength of Wnt signaling in tissues. In this context, R-spondins are secreted proteins that stabilize Wnt receptors and enhance Wnt signaling. In this review we focus on new insights into the regulatory function of Wnt and R-spondin signaling in the stomach. In addition to its function in the healthy state, we highlight the connection between Wnt signaling and infection with Helicobacter pylori (H. pylori), a pathogen that colonizes the stomach and is the main risk factor for gastric cancer. In addition to experimental data that link Wnt signaling to carcinogenesis, we discuss that Wnt signaling is affected in a substantial proportion of patients with gastric cancer, and provide examples for potential clinical implications for altered Wnt signaling in gastric cancer.


Effects of low to moderate levels of deoxynivalenol on feed and water intake, weight gain, and slaughtering traits of broiler chickens.
The aim of the study was to evaluate the effects of low to moderate oral exposure to the Fusarium toxin deoxynivalenol (DON; derived from culture material) on performance, water intake, and carcass parameters of broilers during early and late developmental phases. A total of 160 Ross 308 broilers were randomly allocated to four different feeding groups (n = 40/group) including 0 (control), 2.5, 5, and 10 mg DON/kg wheat-soybean meal-based feed. Three consecutive replicates of the experiment were performed. Half of the broilers were slaughtered in week 3 of the trial whereas the other half were slaughtered in week 5. Dry matter intake (DMI) and water intake (WI) were recorded on a daily basis and the body weight (BW) and BW gain (BWG) were determined weekly. The following carcass traits were recorded and calculated in absolute and relative data: dressed carcass weight, breast muscle weight, leg weight, and liver weight. Data showed that BW (P < 0.001), BWG (P = 0.005), and DMI (P < 0.001) were reduced by DON-feeding during the entire feeding period. The ratio of DMI to body weight gain (DMI/BWG) was not affected by the treatment. However, the ratio of water to DMI (WI/DMI) increased in DON-treated birds (P = 0.021). Contrast analysis showed that DON tendentially reduced slaughter weight (P = 0.082) and decreased leg yield (P = 0.037) in DON-fed chickens in week 5 of the experiment. Liver organ weight decreased in the 3-week-old DON-fed broilers compared to that in the control-fed birds (P = 0.037). In conclusion, the study suggests that DMI and BW were negatively affected under the experimental conditions at DON levels lower than the current guidance value in the European Union of 5 mg/kg feed. The study also indicates that broilers fed on low to moderate level DON-contaminated diets showed increased WI/DMI ratio which might have negative influence on wet litter syndrome.


Acute renal failure in early life.
Data are presented on 25 children under 2 years of age with acute renal failure treated by peritoneal dialysis. The clinical features, treatment, complications, and outcome of the illnesses are described. 22 (88%) survived, and though the incidence of serious complications during the illness was high most regained full health and normal renal function.


A bifunctional Galphai/Galphas modulatory peptide that attenuates adenylyl cyclase activity.
Signaling via G-protein coupled receptors is initiated by receptor-catalyzed nucleotide exchange on Galpha subunits normally bound to GDP and Gbetagamma. Activated Galpha . GTP then regulates effectors such as adenylyl cyclase. Except for Gbetagamma, no known regulators bind the adenylyl cyclase-stimulatory subunit Galphas in its GDP-bound state. We recently described a peptide, KB-752, that binds and enhances the nucleotide exchange rate of the adenylyl cyclase-inhibitory subunit Galpha(i). Herein, we report that KB-752 binds Galpha(s) . GDP yet slows its rate of nucleotide exchange. KB-752 inhibits GTPgammaS-stimulated adenylyl cyclase activity in cell membranes, reflecting its opposing effects on nucleotide exchange by Galpha(i) and Galpha(s).


Genetic analysis of the interaction between cardiac and skeletal actin gene expression in striated muscle of the mouse.
The two sarcomeric actin genes, encoding alpha-cardiac and alpha-skeletal actins, are co-expressed in striated muscle, but in the adult the respective isoform predominates in cardiac or skeletal muscle of the normal mouse. We have investigated the interaction between this gene pair in different genetic contexts. Northern blot analysis of alpha-actin mRNA levels in different inbred mice (129/SJ, C3H, C57BL/6) demonstrates variation of as much as threefold in skeletal muscle and eightfold in cardiac muscle. High or low-level expression is seen for both skeletal and cardiac muscle in a given line, suggesting common regulatory phenomena affecting the abundant alpha-skeletal or alpha-cardiac transcript. In the BALB/c mouse, which has a mutant cardiac actin locus, skeletal as well as cardiac actin mRNA and protein accumulate in the adult heart. We have analysed the role of the two alpha-actin genes in this phenomenon in seven recombinant inbred mouse lines (BALB/c x C57BL/6) and in a cross (BALB/c x C3H). The results demonstrate that neither alpha-actin gene alone is sufficient, and implicate other regulatory loci. DNA sequencing of the C3H and BALB/c alpha-skeletal actin gene promoters shows that they are virtually identical over 830 nucleotides. The relative levels of alpha-skeletal and alpha-cardiac actin proteins have been measured by N-terminal peptide analysis in the different mouse lines. The results point to regulatory loci affecting mRNA utilization and protein stability.


Isolated polycystic liver disease--a rare entity: report of a case.
A 45-year-old woman presented with a mass in the right hypochondrium and shortness of breath. The mass was felt up to 4.5 inches below the right costal margin and its dullness on percussion was continuous with liver dullness. Ultrasonography (USG) of abdomen revealed enlargement of the left lobe of the liver with multiple cysts of varying sizes. Left liver lobectomy was done, histology of which showed multiple cysts lined by cuboidal to columnar epithelium. A small amount amount of normal liver parenchyma between the cysts was observed. A diagnosis of Adult polycystic liver disease (APLD) was given.


Mothers' health care seeking behavior and associated factors for common childhood illnesses, Northwest Ethiopia: community based cross-sectional study.
Substantial progress has been made in reducing child mortality over the last decades, however the magnitude of the problem is yet high globally Appropriate health care-seeking behavior of mothers/guardians for common childhood illnesses could prevent a significant number of child deaths and complications due to childhood illnesses, currently, there is few of studies in Ethiopia. Therefore, this study aimed to assess mothers/caregivers health care seeking behavior for their children in Northwest Ethiopia. Community based cross-sectional study of rural mothers living in Aneded district from February to March 2016. Data were collected using structured questionnaire by an interviewer. Bivariate and multivariable logistic regression analyses were performed to identify factors associated with health care seeking behavior. Variables having P value ≤0.2 in the bivariate analysis were considered for multivariable analysis. P-value less than 0.05 was used to declare that there was statistically significant association. Odds Ratio (OR) with 95% confidence interval (CI) was used to determine the strength and direction of association. A total of 410 mothers participated in this study. Among 48.8% (95% CI: 44, 53.6%) had sought health care, only 27% sought health care within a day. Having awareness of childhood illness (AOR = 3.8, 95% CI: 2.18-6.72), perceived importance of early treatment (AOR = 3.5, 95% CI: 2.00-6.07) and child age < 24 months (AOR = 1.7, 95% CI: 1.08-2.68) and illness not being perceived as severe (AOR:= 0.17, 95% CI: 0.09-0.30) were all factors associated with mothers healthcare seeking behavior during their child illness. Overall health care seeking behavior level was low. Awareness, perceived illness severity, perceived early treatment and having young children were predictors of mothers' health care seeking behavior. The Woreda health office administrators and health professionals should work to improve mothers' awareness and perception towards childhood problems and the importance of early seeking appropriate health care using the existed structures (one-to-five women networking and health developmental army).


Clinical and GAA gene mutation analysis in mainland Chinese patients with late-onset Pompe disease: identifying c.2238G > C as the most common mutation.
Pompe disease is an autosomal recessive lysosomal glycogen storage disorder that has been reported in different ethnic populations which carry different common mutations of the acid alpha-glucosidase (GAA) gene. The GAA mutation pattern in mainland Chinese patients with late-onset Pompe disease is still not well understood. We presented the clinical and genetic characteristics of 27 mainland Chinese late-onset Pompe patients from 24 families. GAA mutation analysis revealed 26 different mutations, including 10 that were novel. The allelic frequency of c.2238G > C (p.W746C) was found to be 27.08% in this patient group. Respiratory dysfunction was diagnosed in 10 of 11 patients who underwent pulmonary function evaluation, although only four required ventilator support at night. Our findings indicate that c.2238G > C (p.W746C) is the most common mutation in mainland Chinese late-onset Pompe patients, as observed in Taiwanese patients. The novel mutations identified in this study expand the genetic spectrum of late-onset Pompe disease, and the prevalence of respiratory dysfunction highlights the importance of monitoring pulmonary function in late-onset Pompe patients.


Omega-3 polyunsaturated fatty acids: benefit or harm during sepsis?
omega-3 polyunsaturated fatty acids (PUFAs) are potent modulators of the immune response. Their inclusion in enteral diets may benefit surgical patients recovering from injury or infection. Caution should be used when supplementing omega-3 PUFAs, particularly when fish oil is used as the source. The long-chain, highly polyunsaturated fatty acids in fish oil are prone to autoxidation, and can potentially damage cells by forming free radicals. In addition, fish oil may impair the hemostatic response by inhibiting platelet aggregation. Finally, the biochemical and biological end points for the use of omega-3 PUFAs have not been clearly established. Thus, although omega-3 PUFAs offer tremendous potential as immunomodulators, they also offer potential toxicities, and we must employ them in our clinical practice with this understanding.


Enteral Nutrition Tolerance And REspiratory Support (ENTARES) Study in preterm infants: study protocol for a randomized controlled trial.
Respiratory distress syndrome (RDS) and feeding intolerance are common conditions in preterm infants and among the major causes of neonatal mortality and morbidity. For many years, preterm infants with RDS have been treated with mechanical ventilation, increasing risks of acute lung injury and bronchopulmonary dysplasia. In recent years non-invasive ventilation techniques have been developed. Showing similar efficacy and risk of bronchopulmonary dysplasia, nasal continuous positive airway pressure (NCPAP) and heated humidified high-flow nasal cannula (HHHFNC) have become the most widespread techniques in neonatal intensive care units. However, their impact on nutrition, particularly on feeding tolerance and risk of complications, is still unknown in preterm infants. The aim of the study is to evaluate the impact of NCPAP vs HHHFNC on enteral feeding and to identify the most suitable technique for preterm infants with RDS. A multicenter randomized single-blind controlled trial was designed. All preterm infants with a gestational age of 25-29 weeks treated with NCPAP or HHHFNC for RDS and demonstrating stability for at least 48 h along with the compliance with inclusion criteria (age less than 7 days, need for non-invasive respiratory support, suitability to start enteral feeding) will be enrolled in the study and randomized to the NCPAP or HHHFNC arm. All patients will be monitored until discharge, and data will be analyzed according to an intention-to-treat model. The primary outcome is the time to reach full enteral feeding, while parameters of respiratory support, feeding tolerance, and overall health status will be evaluated as secondary outcomes. The sample size was calculated at 141 patients per arm. The identification of the most suitable technique (NCPAP vs HHHFNC) for preterm infants with feeding intolerance could reduce gastrointestinal complications, improve growth, and reduce hospital length of stay, thus improving clinical outcomes and reducing health costs. The evaluation of the timing of oral feeding could be useful in understanding the influence that these techniques could have on the development of sucking-swallow coordination. Moreover, the evaluation of the response to NCPAP and HHHFNC could clarify their efficacy as a treatment for RDS in extremely preterm infants. ClinicalTrials.gov, NCT03548324 . Registered on 7 June 2018.


Endocrine regulation of periparturient behaviour in pigs.
Pigs begin behavioural preparations for birth about 1-2 days before parturition. Prepartum sows wander to select a suitable site and then construct a maternal nest. The signal that initiates this behavioural cascade probably results from fetal maturation but is unknown. However, endogenous PGF2 alpha appears to be involved early on in an endocrine pathway that projects to the brain and can generate most of the prepartum behavioural components. This period of intense activity is followed by a quiescent phase of lying in the nest for some hours before fetal ejection occurs. Feedback from a completed nest or abdominal discomfort may both contribute to the end of nest building. In the postpartum phase, sows have to deal with the apparently conflicting drives of remaining passive to reduce accidental or deliberate damage to piglets, while at the same time responding actively to their needs. In commercial environments, animals frequently fail in this task. Although environmental influences on piglet survival have received much experimental attention, the genetic, social and endocrine drives that control sow behaviour after parturition remain poorly understood and their clarification is a major challenge for the future.