The state of art of neutrophil extracellular traps in protozoan and helminthic infections.
Neutrophil extracellular traps (NETs) are DNA fibers associated with histones, enzymes from neutrophil granules and anti-microbial peptides. NETs are released in a process denominated NETosis, which involves sequential steps that culminate with the DNA extrusion. NETosis has been described as a new mechanism of innate immunity related to defense against different pathogens. The initial studies of NETs were carried out with bacteria and fungi, but currently a large variety of microorganisms capable of inducing NETs have been described including protozoan and helminth parasites. Nevertheless, we have little knowledge about how NETosis process is carried out in response to the parasites, and about its implication in the resolution of this kind of disease. In the best case, the NETs entrap and kill parasites in vitro, but in others, immobilize the parasites without affecting their viability. Moreover, insufficient studies on the NETs in animal models of infections that would help to define their role, and the association of NETs with chronic inflammatory pathologies such as those occurring in several parasitic infections have left open the possibility of NETs contributing to pathology instead of protection. In this review, we focus on the reported mechanisms that lead to NET release by protozoan and helminth parasites and the evidence that support the role of NETosis in the resolution or pathogenesis of parasitic diseases.


Snake Cathelicidin Derived Peptide Inhibits Zika Virus Infection.
Zika virus (ZIKV) is a mosquito-borne virus belonging to the genus Flavivirus and has reemerged in recent years with epidemic potential. ZIKV infection may result in severe syndromes such as neurological complications and microcephaly in newborns. Therefore, ZIKV has become a global public health threat and currently there is no approved specific drug for its treatment. Animal venoms are important resources of novel drugs. Cathelicidin-BF (BF-30) is a defensive peptide identified from Bungarus fasciatus snake venom and has been shown to be an excellent template for applicable peptide design. In this study, we found that ZY13, one of the peptidic analogs of BF-30, inhibits ZIKV infection in vitro and in vivo. Mechanistic studies revealed that ZY13 can directly inactivate ZIKV and reduce the production of infectious virions. Further studies also indicated that administration of ZY13 strengthen the host antiviral immunity via AXL-SOCS (suppressor of cytokine signaling protein) pathway. Additionally, the results of mouse experiment suggest that ZY13 efficiently restrict ZIKV infection and improve the growth defects of ZIKV-infected mouse pups. Together, our findings not only demonstrate that ZY13 might be a candidate for anti-ZIKV drug, but also indicated the importance of animal venom peptides as templates for antivirals development.


Community genetics in the Northwestern Atlantic intertidal.
Our ability to make inferences about the processes acting upon a biological system can be dramatically improved through integration of information from other fields. In particular, this is true of the field of phylogeography, a discipline that attempts to describe geographical variation in species using neutral genetic diversity as a correlate of time. Through comparisons of information from multiple species, as well as background information about the abiotic environment and how it has changed over time, we should be able to reassemble many of the forces that were important in assembling the communities and community interactions found in a given region. Here I review the information available for coastal species in the northwestern Atlantic, and argue that an integration of ecological, genetic, geological and oceanographic information can illustrate emergent patterns of community genetics.


Highly brominated antimicrobial metabolites from a marine Pseudoalteromonas sp.
Extracts of a marine Pseudoalteromonas sp. (CMMED 290) isolated from the surface of a nudibranch collected in Kaneohe Bay, Oahu, displayed significant antimicrobial activity against methicillin-resistant Staphylococcus aureus. Bioassay-guided fractionation of the lipophilic extract led to the isolation and structure elucidation of two new highly brominated compounds, 2,3,5,7-tetrabromobenzofuro[3,2-b]pyrrole (1) and 4,4',6-tribromo-2,2'-biphenol (2). In addition, we have identified the known compounds pentabromopseudilin and bromophene. We describe the isolation and structure elucidation of the compounds 1 and 2 together with their antimicrobial activities against methicillin-resistant Staphylococcus aureus.


Mutation of the important Tyr-33 residue of chicken avidin: functional and structural consequences.
The strong interaction between avidin and biotin is so tight (dissociation constant 10(-15) M) that conditions usually sufficient for protein denaturing fail to dislodge biotin from the avidin-biotin complex. This kind of irreversible binding hinders the use of avidin in applications such as affinity purification or protein immobilization. To address this concern, we have constructed a series of mutants of the strategically positioned Tyr-33 in order to study the role of this residue in biotin binding, and to create avidin variants with more reversible ligand-binding properties. Unexpectedly, an avidin mutant in which Tyr-33 was replaced with phenylalanine (Avm-Y33F) displayed similar biotin-binding characteristics to the native avidin, indicating that the hydrogen bond formed between the hydroxy group of Tyr-33 and the carbonyl oxygen of biotin is not as important for the tight binding of biotin as previously suggested. In terms of the reversibility of biotin binding, Avm-Y33H was the most successful substitution constructed in this study. Interestingly, the binding of this mutant exhibited clear pH-dependence, since at neutral pH it bound to the biotin surface in an irreversible fashion, whereas, at pH 9, 50% of the bound protein could be released with free biotin. Furthermore, although Tyr-33 is located relatively distant from the monomer-monomer interfaces, the mutagenesis of this residue also weakened the quaternary structure of avidin, indicating that the high ligand binding and the high stability of avidin have evolved together and it is difficult to modify one without affecting the other.


Neuroanatomy of hemispatial neglect and its functional components: a study using voxel-based lesion-symptom mapping.
Spatial neglect is a perplexing neuropsychological syndrome, in which patients fail to detect (and/or respond to) stimuli located contralaterally to their (most often right) hemispheric lesion. Neglect is characterized by a wide heterogeneity, and a role for multiple components has been suggested, but the exact nature of the critical components remains unclear. Moreover, many different lesion sites have been reported, leading to enduring controversies about the relative contribution of different cortical and/or subcortical brain regions. Here we report a systematic anatomo-functional study of 80 patients with a focal right hemisphere stroke, who were examined by a series of neuropsychological tests assessing different clinical manifestations of neglect. We first performed a statistical factorial analysis of their behavioural performance across all tests, in order to break down neglect symptoms into coherent profiles of co-varying deficits. We then examined the neural correlates of these distinct neglect profiles using a statistical voxel-based lesion-symptom mapping method that correlated the anatomical extent of brain damage with the relative severity of deficits along the different profiles in each patient. Our factorial analysis revealed three main factors explaining 82% of the total variance across all neglect tests, which suggested distinct components related to perceptive/visuo-spatial, exploratory/visuo-motor, and allocentric/object-centred aspects of spatial neglect. Our anatomical voxel-based lesion-symptom mapping analysis pointed to specific neural correlates for each of these components, including the right inferior parietal lobule for the perceptive/visuo-spatial component, the right dorsolateral prefrontal cortex for the exploratory/visuo-motor component, and deep temporal lobe regions for the allocentric/object-centred component. By contrast, standard anatomical overlap analysis indicated that subcortical damage to paraventricular white matter tracts was associated with severe neglect encompassing several tests. Taken together, our results provide new support to the view that the clinical manifestations of hemispatial neglect might reflect a combination of distinct components affecting different domains of spatial cognition, and that intra-hemispheric disconnection due to white matter lesions might produce severe neglect by impacting on more than one functional domain.


The physiology of lymphocyte migration through the single lymph node in vivo.
Lymphocytes are mobile cells, continually recirculating between the blood and the tissues via the lymph. In order to maintain immune surveillance, the majority of lymphocyte traffic occurs through lymph nodes in vivo. Although a great deal of work has been done to elucidate the molecular mechanisms whereby lymphocytes leave the blood and enter the lymph node, lymphocyte traffic also requires that the lymphocyte successfully transit extravascular tissue and enter the lymph following transendothelial migration. The regulation of cell movement through lymph nodes, specific cellular positioning within the nodes, and eventual entry into the efferent lymphatics are poorly understood. The process of lymphocyte recirculation occurs in a physiological background, and in vivo systems have been particularly useful in uncovering the nuances of the process. This review summarizes available data about the recirculation of lymphocytes through the lymph node and the interaction of recirculating lymphocyte pools in vivo. The importance of factors in afferent lymph, the specific distribution of extracellular matrix proteins, potential soluble regulators of cell traffic, and evidence for an active role of lymphatic endothelial cells in the regulation of lymphocyte traffic are discussed. It seems likely that future work will need to be directed at determining the relative importance of these post-transendothelial migration regulators of lymphocyte traffic.


Malignant giant pheochromocytoma: a case report and review of the literature.
Malignant pheochromocytoma is a rare disease and surgical resection is the only curative treatment. There are no definitive histological or cytological criteria of malignancy, as it is impossible to determine this condition in the absence of advanced locoregional disease or metastases. We report a case of a patient with a giant retroperitoneal tumour, the second largest to be published, which was diagnosed as a malignant pheochromocytoma; it was treated with surgery. The literature is reviewed to evaluate tumour features and criteria to distinguish between benign and malignant pheochromocytomas.


Higher frequency of point mutations in the c-K-ras 2 gene in human colorectal adenomas with severe atypia than in carcinomas.
Human colorectal carcinomas may be induced from adenomas or they may occur de novo. To examine which is the main pathway, we analyzed point mutations at codon 12 in the c-K-ras 2 gene in 73 colorectal carcinomas, 13 metastatic tumors, 72 adenomas and 30 normal tissues. The c-K-ras 2 codon 12 mutation frequency was 0/30 in normal tissues, 0/17 in adenomas with mild atypia, 3/37 (8.1%) in adenomas with moderate atypia, 15/18 (83.3%) in adenomas with severe atypia, 19/73 (26.0%) in primary carcinomas and 3/13 (23.1%) in metastatic tumors. The mutation frequency in adenomas with severe atypia was much higher than that in carcinomas. These results indicate that many colorectal carcinomas may not be induced through adenomas with severe atypia.


Structural modeling and electron paramagnetic resonance spectroscopy of the human Na+/H+ exchanger isoform 1, NHE1.
We previously presented evidence that transmembrane domain (TM) IV and TM X-XI are important for inhibitor binding and ion transport by the human Na(+)/H(+) exchanger, hNHE1 (Pedersen, S. F., King, S. A., Nygaard, E. B., Rigor, R. R., and Cala, P. M. (2007) J. Biol. Chem. 282, 19716-19727). Here, we present a structural model of the transmembrane part of hNHE1 that further supports this conclusion. The hNHE1 model was based on the crystal structure of the Escherichia coli Na(+)/H(+) antiporter, NhaA, and previous cysteine scanning accessibility studies of hNHE1 and was validated by EPR spectroscopy of spin labels in TM IV and TM XI, as well as by functional analysis of hNHE1 mutants. Removal of all endogenous cysteines in hNHE1, introduction of the mutations A173C (TM IV) and/or I461C (TM XI), and expression of the constructs in mammalian cells resulted in functional hNHE1 proteins. The distance between these spin labels was ∼15 A, confirming that TM IV and TM XI are in close proximity. This distance was decreased both at pH 5.1 and in the presence of the NHE1 inhibitor cariporide. A similar TM IV·TM XI distance and a similar change upon a pH shift were found for the cariporide-insensitive Pleuronectes americanus (pa) NHE1; however, in paNHE1, cariporide had no effect on TM IV·TM XI distance. The central role of the TM IV·TM XI arrangement was confirmed by the partial loss of function upon mutation of Arg(425), which the model predicts stabilizes this arrangement. The data are consistent with a role for TM IV and TM XI rearrangements coincident with ion translocation and inhibitor binding by hNHE1.


[Fused TU complexes as a new electrocardiographic marker of poor prognosis after myocardial infraction].
To date, the clinical significance of fused T and U waves, termed as TU complexes, has not been evaluated. The aim of the study was to present the clinical characteristics of the patients with TU complexes and to assess the value of this ECG abnormality in risk stratification after myocardial infarction. In the group of 330 postinfarction patients (mean age 61 +/- 10 years, 279 men and 51 women), 50 (15%) had TU complexes in one or more leads of a standard ECG. In patients with TU complexes, the decreased left ventricular ejection fraction, frequent (> or = 10/hour) ventricular premature beats and non-sustained ventricular tachycardia detected on 24-hours ECG monitoring, increased QT dispersion, ST-segment depression and ST-segment elevation on a routine ECG were more common than in patients without TU complexes. During a follow-up period of 43 +/- 17 months, 88 patients died from all causes. At univariate Cox analysis the presence of TU complexes (hazard ratio 3.30; 95% confidence interval 2.09-5.21) and left ventricular ejection fraction < 40% (hazard ratio 3.82; 95% confidence interval 2.51-5.82) were the best predictors of mortality among the 9 evaluated clinical and electrocardiographic variables. The multivariate, stepwise Cox analysis selected ejection fraction < 40% (hazard ratio 3.09; 95% confidence interval 2.00-4.80), TU complexes (hazard ratio 2.28; 95% confidence interval 1.42-3.69), RR interval < 800 ms (hazard ratio 1.62; 95% confidence interval 1.06-2.47), and age of patients > 65 years (hazard ratio 1.58; 95% confidence interval 1.03-2.42) as an independent predictors of all cause mortality. The presence of TU complexes on a routine ECG is associated with impaired left ventricular function, increased predisposition to ventricular arrhythmias and higher risk of mortality. TU complexes may be considered as a new electrocardiographic marker of poor prognosis in patients after myocardial infarction.


Detection of an IS21 insertion sequence in the mexR gene of Pseudomonas aeruginosa increasing beta-lactam resistance.
To understand the regulation of the MexAB OprM efflux system in a clinical strain of Pseudomonas aeruginosa presenting a decreased susceptibility to ticarcillin and aztreonam, the mexR repressor gene was amplified by polymerase chain reaction (PCR) and was shown to be disrupted by an insertion sequence of more than 2 kb, with characteristic direct and inverted repeat sequences. Sequencing revealed a 2131-bp IS21 insertion sequence. A reverse transcription PCR method was used to quantify mexA transcripts and showed an increased transcription rate of mexA in this strain, compared with a PAO1 control strain. The nalB phenotype in P. aeruginosa may be due to point mutations, but also to the presence of an insertion sequence in the mexR regulator gene.


New UDP-GlcNAc C4 epimerase involved in the biosynthesis of 2-acetamino-2-deoxy-L-altruronic acid in the O-antigen repeating units of Plesiomonas shigelloides O17.
Plesiomonas shigelloides is a ubiquitous waterborne pathogen responsible for diseases such as diarrhea and bacillary dysentery, commonly afflicting infants and children. This bacterium is endowed with an O-antigen gene cluster consisting of 10 consecutive reading frames. One of these, designated wbgU (orf3), has been overexpressed and biochemically characterized to show that it encodes a uridine diphosphate-N-acetylglucosamine (UDP-GlcNAc) C4 epimerase, only the second microbial enzyme characterized to have this activity. Epimerization is an equilibrium reaction resulting in a 70:30 ratio of UDP-GlcNAc to uridine diphosphate-N-acetylgalactosamine (UDP-GalNAc), irrespective of the initial substrate. The K(m) values for UDP-GalNAc and UDP-GlcNAc are 131 microM and 137 microM, respectively. WbgU is also capable of converting nonacetylated derivatives but with much lower efficiency. It contains a tightly bound nicotinamide adenine dinucleotide [NAD(H)] molecule and requires no other cofactors for activity. We propose here that this enzyme catalyzes the first of the three transformations in the biosynthetic pathway of 2-acetamino-2-deoxy-L-altruronic acid, an unusual sugar present in the O-specific side chains of lipopolysaccharide of P. shigelloides O17 and its close relative Escherichia coli Sonnei.


[Management, prevention and control of pernicious anemia].
Pernicious anemia is the most frequent cause of megaloblastic anemia in our area, and it is the result of a vitamin B12 deficiency due, itself, to the decrease or absence of intrinsic factor (IF) because of gastric mucosa atrophy or autoimmune destruction of IF-producing parietal cells. With the existence of a severe gastric atrophy, there is a decrease in acid and IF production and a further change in vitamin B12 absorption. Fifty percent of the cases are associated to anti-IF antibodies, which presence in other autoimmune diseases is exceptional. In patients with pernicious anemia, measurement of anti-IF antibodies has high specificity (95%); however, measurement of anti-parietal cells antibodies has low specificity. The first-choice treatment is administration of vitamin B12 intramuscularly. The regimen is the administration of 1 mg of vitamin B12 daily for one week, weekly thereafter for one month and, then, every 2-3 months for life.


Pharmacokinetics of rapamycin-eluting stents in miniswine coronary model.
The results of clinical trials of rapamycin-eluting stents reduce restenosis have been quite promising. The main purpose of this study was to characterize the in vivo pharmacokinetics of high dose rapamycin (Rapa)-eluting stents in a miniswine coronary model. Ten miniswines underwent placement of 18 high dose Rapa-eluting stents in the left anterior descending and right coronary arteries. At the planned times of the 1.5th, 12th, 24th hour, 3th, 7th and 28th day, the animals (n = 1, 1, 2, 2, 2, and 2, respectively) were euthanized after completion of coronary angiography. Blood samples were obtained at 0, 10, 20, 30 minutes; 1, 2, 6, 24 hours; and 3, 7, 28 days to determine systemic Rapa levels. Rapa levels in whole blood, arterial wall, heart, renal and liver tissues were determined by high-performance liquid chromatography/mass spectroscopy. Peak whole blood concentration (Cmax), time to peak concentration (tmax), elimination half-life (t1/2beta), area under the curve (AUC), and apparent systemic clearance (Cl/F) were (10.91 +/- 1.28) ng/ml, (2.0 +/- 0.2) hours, (7.25 +/- 0.63) hours, (1.15 +/- 0.11) ng x h x ml(-1), and (180 +/- 12) ml x h(-1) x kg(-1), respectively. More than 95% Rapa detected is localized in the coronary artery surrounding the stent and heart. Stent-based delivery of Rapa via a copolymer stent is feasible and safe. This strategy holds promise for the prevention of stent restenosis.


A systematic review and evaluation of response redirection as a treatment for challenging behavior in individuals with developmental disabilities.
Response redirection is widely used in clinical practice as a treatment for repetitive behavior or stereotypy in persons with developmental disabilities. However, to date the procedure has received comparatively little empirical evaluation. The current review sought to examine the literature describing the efficacy of response redirection alone, response interruption and redirection (RIRD), and multi-element treatment packages incorporating response redirection, as interventions for challenging behavior in individuals with developmental disabilities. Additionally, the status of response redirection, and RIRD, as evidence-based practice was evaluated in accordance with Reichow's (2011) recently developed criteria. Results indicated that interventions involving response redirection or RIRD typically led to large decreases in challenging behavior but did not result in behavioral suppression. On the basis of the current literature and in accordance with Reichow's criteria, interventions incorporating response redirection do not yet constitute evidence-based practice. The implications of these findings, for both research and practice, are discussed.


[The radiological study on the open reduction of congenital dislocation of the hip by Ludloff's method. Cases more than 5 years after operation].
Fourty-eight patients with fifty-seven congenitally dislocated hips that had been treated surgically using the Ludloff's technique were followed up for an average of five years from the time of operation. At the final follow up examination the radiological results were good in sixty-five per cent of the hips, fair in twenty-eight per cent and poor in seven per cent. As relating to the age of operation, it was found that good results were seen in eighty-two per cent of the patients below the age of one year, comparing with fourty-eight per cent seen in those patients aged over one year. The incidence of avascular necrosis was fourteen per cent. Although coxa magna was noted in seventy-nine per cent of the hips, there has been a low incidence of coxa magna in those hips in which the ligamentum teres were preserved during operation. About one third of the congenitally dislocated hips in this study required salvage operation after open reduction. In order to find out a reference which would be available in predicting the prognosis of open reduction, a retrospective study of X-ray findings was made. The author found that the prognosis was favorable in those hips which had the CE angle over ten degrees at the time when the brace was removed, whereas a majority of the hips in which the CE angle were below zero needed salvage operation. It was also found that the prognosis of open reduction was good in those patients who had the capital-teardrop distance shorter than ten milimeters and the acetabular head index over sixty per cent at the age of three to four years.


The effect of dietary fat saturation on plasma and hepatic lipoproteins in the rat.
Semipurified diets containing 10% kilocalories from either safflower oil (SO), corn oil (CO), olive oil (OO) or palm oil (PO) were fed to weanling male rats for 2 weeks. The effects of dietary fat saturation on plasma lipids and lipoproteins were: 1) Nonfasted plasma cholesterol concentration was higher in rats fed OO (mean +/- SEM = 81.0 +/- 2.9 mg/dl) vs. CO (67.5 +/- 2.9); 2) plasma chylomicron cholesterol concentration was higher in rats fed OO vs. SO and CO, with PO values in between; and 3) the cholesterol concentration of plasma very low density lipoprotein (VLDL), low density lipoprotein (LDL) and high density lipoprotein (HDL) did not differ among groups. The effects of dietary fat saturation on hepatic lipoproteins (determined by liver perfusion techniques) were: 1) hepatic higher density lipoprotein (d = 1.006-1.21 g/ml) cholesterol production was greater in rats fed SO and CO vs. PO [19.1 +/- 1.2, 17.2 +/- 0.8 and 13.7 +/- 1.6 micrograms/(g liver X 1.5 hour), respectively]; 2) there was no difference in hepatic VLDL cholesterol production among groups; and 3) the ratio of cholesterol to protein of hepatic VLDL and the higher density lipoprotein fraction was higher in rats fed diets rich in polyunsaturated fatty acids versus saturated fatty acids. Dietary fat saturation had no effect on carcass and liver cholesterol concentrations. Since differences in hepatic lipoprotein production were not reflected in plasma lipoprotein patterns, these results suggest that extrahepatic lipoprotein metabolism differs in rats fed diets containing fatty acids of varying saturation.


The electrocardiogram as a prognostic tool for predicting major cardiac events.
Among patients with cardiac disease, the identification of those who are at low risk and those who are at high risk for major cardiac events is crucial for a rational clinical management of individual patients. A correct noninvasive risk stratification of cardiac patients, in particular, has relevant clinical implications because it would avoid unnecessary exposure to potentially risky invasive diagnostic or interventional procedures in low-risk patients, whereas it would allow an appropriate aggressive diagnostic and therapeutic approach in high-risk patients. Furthermore, the appropriate identification of low- and high-risk patients would also have social and economic implications by favoring optimization of resource distribution and costs. A large number of studies in previous decades provided evidence that several methods and variables derived from the analysis of the electrocardiogram (ECG) are powerful predictors of major cardiac events in several clinical conditions. Despite that, there has been limited attention about how several of these findings can be used in clinical practice. Furthermore, in recent years, most studies about risk stratification of cardiac patients have mainly been focused on the use of a number of serum/plasma biomarkers with reduced attention to ECG variables. Surprisingly, however, there have been few attempts to establish whether the various proposed risk markers add any significant information to that obtainable from ECG methods. In this article, the evidence for the prognostic value of variables derived from the assessment of the ECG signal by several methods and techniques will be briefly reviewed. Because of the largeness of the topic, this review will be necessarily incomplete. Because most of the clinical research in this field concerned risk stratification of patients with coronary artery disease, the article will be largely focused on this population of patients. The role of ECG methods in specific cardiac diseases and, in particular, in the general population of asymptomatic subjects will be briefly discussed when believed appropriate and helpful. Furthermore, only major clinical events (ie, cardiac death, arrhythmic events, acute myocardial infarction) will be taken into account as end points in this article. Minor clinical events (eg, coronary revascularization procedures, coronary artery restenosis, recurrences of symptoms) are indeed less robust as end points because they are widely biased by subjective judgments.


Morphological aspects of the vasculogenesis and angiogenesis during prenatal edification of the circle of Willis: a review.
In the literature, there are many articles reporting anatomical variations of circle of Willis (CoW), defined as those changes that lead to the inability of this anastomotic structure to maintain adequate brain flow. Because there is such a wide variation in the configuration of the CoW, its anatomical variations affect the hemodynamics of blood flow, thus contributing to the development of aneurysms or stroke. As such, we consider that a good knowledge of the embryological development of the constituent arteries of the CoW can shed some light on the causes of the appearance of its anatomical variants. Reviewing literature, we will present the embryological development of the constituting arteries of the CoW and will begin with vasculogenesis and angiogenesis of the vascular system as a whole. Then, we will focus on the embryological development of the internal carotid artery (ICA) and its branches because, starting with the embryological day 24, these arteries are the first vessels that begin to develop to provide the necessary blood for the primitive brain. As the hindbrain increases its volume, a larger amount of nutrients is needed. Because a larger amount of blood is required to be provided by the primitive ICAs, there is a need for arterial capacity development and thus the posterior circulation begin to take shape. At this stage, the posterior circulation consists of a plexiform arterial network that receives blood from the carotid artery through the carotid-vertebrobasilar anastomoses. At the 5-8 mm embryonic stage, these anastomoses begin to regress, and the basilar artery and vertebral arteries become independent of the ICA. We are pointing out on the process of regression of these primitive vessels, emphasizing the fact that their persistence represents the starting point for the appearance of anatomic anomalies of the CoW, which are identified in the adult individuals. In this review, we also present and illustrate some developmental abnormalities of the anterior and posterior parts of the CoW.


Quantitative Proteomics Reveals Common and Specific Responses of a Marine Diatom Thalassiosira pseudonana to Different Macronutrient Deficiencies.
Macronutrients such as nitrogen (N), phosphorus (P), and silicon (Si) are essential for the productivity and distribution of diatoms in the ocean. Responses of diatoms to a particular macronutrient deficiency have been investigated, however, we know little about their common or specific responses to different macronutrients. Here, we investigated the physiology and quantitative proteomics of a diatom Thalassiosira pseudonana grown in nutrient-replete, N-, P-, and Si-deficient conditions. Cell growth was ceased in all macronutrient deficient conditions while cell volume and cellular C content under P- and Si-deficiencies increased. Contents of chlorophyll a, protein and cellular N decreased in both N- and P-deficient cells but chlorophyll a and cellular N increased in the Si-deficient cells. Cellular P content increased under N- and Si-deficiencies. Proteins involved in carbon fixation and photorespiration were down-regulated under all macronutrient deficiencies while neutral lipid synthesis and carbohydrate accumulation were enhanced. Photosynthesis, chlorophyll biosynthesis, and protein biosynthesis were down-regulated in both N- and P-deficient cells, while Si transporters, light-harvesting complex proteins, chloroplastic ATP synthase, plastid transcription and protein synthesis were up-regulated in the Si-deficient cells. Our results provided insights into the common and specific responses of T. pseudonana to different macronutrient deficiencies and identified specific proteins potentially indicating a particular macronutrient deficiency.


Contribution of VEGF polymorphisms to variation in VEGF serum levels in a healthy population.
Vascular endothelial growth factor (VEGF) is a pro-angiogenic factor. Variability in VEGF expression, induced by specific VEGFA variants, are involved in angiogenesis-related disorders. This study examined the genotype distribution and functional role (VEGF expression) of rs699947, rs833061, rs1570360, rs2010963, rs833068, rs833070, rs3025020, and rs3025039 VEGFA variants and their haplotypes in 519 healthy Bahraini individuals of both genders. The distribution of the eight VEGFA polymorphisms screened was in Hardy-Weinberg equilibrium. The minor allele frequencies of rs699947 (0.42), rs833061 (0.32), rs1570360 (0.31), rs2010963 (0.33), rs833068 (0.37), rs833070 (0.42), rs3025020 (0.33), and rs3025039 (0.13) were generally compared to those established for Caucasians. Of the variants tested, rs3025020 was associated with increased VEGF serum levels (p=0.019), while rs3025039 was associated with decreased levels (p=0.038). Linkage analysis identified two VEGFA blocks, the first, spanning 16 kb, was not associated with altered VEGF levels, while the second, spanning 3 kb containing rs3025020 and rs3025039, was linked with higher VEGF expression, of which the (-583)T/(+936)T haplotype (p=0.008) was linked with higher VEGF levels compared to the (-583)C/(+936)C (all wild-type) haplotype. These results support the association of rs30250202 and rs3025039, and specific VEGF haplotypes, with altered VEGF serum levels, although the exact functional mechanisms remain to be elucidated.


Detection of hepatocellular carcinoma in cirrhotic patients: sensitivity of CT and ultrasonography.
Patients with cirrhosis are at increased risk for the development of hepatocellular carcinoma. The heterogeneous hepatic parenchyma produced by cirrhosis makes detection of hepatomas more difficult. The purpose of this study was to determine the sensitivities of CT and ultrasonography for detecting hepatomas in cirrhotic patients. A retrospective analysis was performed of 733 patients who underwent liver transplantation at our institution. A study population of 21 patients was selected who met our inclusion criteria. The inclusion criteria required a pathologic diagnosis of hepatocellular carcinoma, pathologic evidence of cirrhosis, and contrast-enhanced CT and sonographic examinations performed within 1 week of each other. The sensitivities of CT and ultrasonography were determined by comparing the imaging findings with pathology findings from serially sectioned total hepatectomy specimens. A total of 40 hepatomas were detected pathologically in the 21 patients in our study population. CT identified 12 of 21 patients with hepatomas and detected 18 of 40 individual lesions (patient detection sensitivity = 57%, lesion detection sensitivity = 45%). Ultrasonography identified 14 of 21 patients with hepatomas and detected 21 of 40 individual lesions (patient detection sensitivity = 67%, lesion detection sensitivity = 51%). Combining the findings of CT and ultrasonography allowed identification of 17 of 21 patients with hepatomas and detection of 24 of 40 individual lesions (patient detection sensitivity = 80%, lesion detection sensitivity = 60%). We conclude that CT and ultrasonography have a low sensitivity for the detection of hepatocellular carcinoma in patients with cirrhosis.


Microenvironmental stimuli induce different macrophage polarizations in experimental models of emphysema.
Macrophages play a pivotal role in the development of emphysema and depending on the microenvironment stimuli can be polarized into M1- or M2-like macrophage phenotypes. We compared macrophage polarizations in cigarette smoke (CS)- and porcine pancreatic elastase (PPE)-induced emphysema models. C57BL/6 mice were subdivided into four experimental groups. In the PPE group, animals received an intranasal instillation of PPE (0.677 IU); in the saline group, animals received an intranasal instillation of saline (0.9%). Animals from both groups were euthanized on day 28. In the CS group, animals were exposed to CS for 30 min, twice a day, 5 days per week for 12 weeks. In the control group, animals received filtered air. We observed an increase in total macrophages for both experimental models. For M1-like macrophage markers, we observed an increase in TNF-α+ and IFN-γ+ cells, Cxcl-9 and Cxcl-10 expressions in PPE and CS groups. Only in the CS group, we detected an increased expression of IL-12b For M2-like macrophages markers we observed a down regulation in IL-10, IL-4, IL-13, Arg1 and Fizz1 and an increase of TGF-β+ cells in the PPE group, while for the CS group there was an increase in TGF-β+ cells and IL-10 expression. All exposure groups were compared to their respective controls. In summary, we demonstrated that CS- and PPE-induced models resulted in different microenvironmental stimuli. CS exposure induced an environmental stimulus related to M1- and M2-like macrophage phenotypes similar to previous results described in COPD patients, whereas the elastase-induced model provided an environmental stimulus related only to the M1 phenotype.


[Advancement of imaging technology for coronary microcirculation dysfunction assessment].
Coronary microcirculation dysfunction (CMVD) is an important risk factor for the prognosis of re-perfused ischemic heart. Recent studies showed that the evaluation of CMVD has significant impact on both the early diagnosis of heart diseases relevant to blood supply and prognosis after myocardial reperfusion. In this review, the definition of CMVD from the perspective of pathophysiology was clarified, the principles and features of the state-of-the-art imaging technologies for CMVD assessment were reviewed from the perspective of engineering and the further research direction was promoted.


Morphine-induced conditioned place preference in rhesus monkeys: Resistance to inactivation of insula and extinction.
Drug addicts experience strong craving episodes in response to drug-associated cues. Attenuating these responses using pharmacological or behavioral approaches could aid recovery from addiction. Cue-induced drug seeking can be modeled using the conditioned place preference procedure (CPP). Our previous work showed that conditioned place preference (CPP) can be induced by administration of increasing doses of morphine in rhesus monkeys. Here, we investigated whether expression of morphine-induced CPP can be attenuated by inhibiting activity of insular cortex or by repeated unreinforced exposures to the CPP test. The insula has been demonstrated to be involved in addiction to several drugs of abuse. To test its role in morphine CPP, bilateral cannulae were implanted into the insula in seven adult monkeys. The CPP was established using a biased apparatus by intramuscular injections of morphine at increasing doses (1.5, 3.0 and 4.5mg/kg) for each monkey. After the monkeys established morphine CPP, their insulae were reversibly inactivated by bilateral microinjection with 5% lidocaine (40μl) prior to the post-conditioning test (expression) of CPP using a within-subject design. The microinjections of lidocaine failed to affect CPP expression when compared to saline injections. We subsequently investigated morphine-associated memory during six episodes of CPP tests performed in these monkeys over the following 75.0±0.2months. While the preference score showed a declining trend with repeated testing, morphine-induced CPP was maintained even on the last test performed at 75months post-conditioning. This observation indicated strong resistance of morphine-induced memories to extinction in rhesus monkeys. Although these data do not confirm involvement of insula in morphine-induced CPP, our observation that drug-associated memories can be maintained over six drug-free years following initial experience with morphine has important implications for treatment of drug addiction using extinction therapy.


Health status utilities in pediatrics: a systematic review of acute lymphoblastic leukemia.
Measuring utilities and health-related quality of life (HRQL) in children is challenging due to their cognitive abilities and changing developmental stages. . To identify methodological issues on utility measurements in children, we performed a systematic review on utilities measured with a single instrument, the Health Utilities Index (HUI), in pediatric acute lymphoblastic leukemia (ALL). The secondary goal was to facilitate future cost-utility analyses without the need for time-consuming assessments. Data Sources. PubMed, Embase, Cochrane Library, CINAHL, and PsycINFO were searched from inception to June 2012. Studies had to report on utility scores in pediatric ALL, either on or after treatment, to be included. . Fifteen studies were included. Most studies had methodological shortcomings, which mainly concerned and definition and representativeness of the study group. Utility scores were dependent on treatment variables, and there generally was an improvement in HRQL as treatment or survivorship advanced. In general, proxy-respondents were less reliable for subjective phenomena than for observable conditions. HUI2 and HUI3 scores were not interchangeable. Limitations. Studies may have been missed because no validated search method for utility studies exists, due to language bias or the exclusion of non-peer-reviewed papers. . Most studies in this review were methodologically suboptimal. Future developments should focus on including developmentally appropriate items for the whole pediatric age group. Adding disease-specific domains may enhance the sensitivity and responsiveness of instruments. Efforts should be undertaken to elicit valuation of health states from older children and teenagers as much as possible. For now, it remains difficult to make valid and informed decisions on the financing of interventions until health state valuation in children has become more methodologically robust.


Neurosurgery and the dawning age of Brain-Machine Interfaces.
Brain-machine interfaces (BMIs) are on the horizon for clinical neurosurgery. Electrocorticography-based platforms are less invasive than implanted microelectrodes, however, the latter are unmatched in their ability to achieve fine motor control of a robotic prosthesis capable of natural human behaviors. These technologies will be crucial to restoring neural function to a large population of patients with severe neurologic impairment - including those with spinal cord injury, stroke, limb amputation, and disabling neuromuscular disorders such as amyotrophic lateral sclerosis. On the opposite end of the spectrum are neural enhancement technologies for specialized applications such as combat. An ongoing ethical dialogue is imminent as we prepare for BMI platforms to enter the neurosurgical realm of clinical management.


Full longitudinal nailfold videocapillaroscopy analysis of microvascular changes during normal pregnancy.
Microvascular remodeling is one major responsible for vascular adaptation in pregnancy, still it is not routinely evaluated in the obstetric field. This pilot study aimed to explore the role of nailfold capillaroscopy (NCV) in detecting microvascular changes during normal pregnancy. A population of 30 healthy pregnant women was longitudinally followed performing clinical assessment and NVC evaluation at each trimester and post-partum. Thirty non-pregnant age-matched healthy women having received at least two NVCs with a minimum 9 to 12-month interval were selected as controls. All NVC images were evaluated by a qualitative and semi-quantitative assessment using current standardised approach. Statistical analyses were conducted to assess NVC trend throughout gestation and its possible association with pregnancy course. A progressive significant increase of NVC neoangiogenesis and a specular reduction in capillary dilations was observed during pregnancy (p < 0.05). These variations were not found in age-matched controls, who showed stable NVC parameters over a similar time frame (p < 0.05). Additionally, a significant inverse correlation was found between NVC neoangiogenesis rate and maternal systemic BP (rho = -0.72, p < 0.005). This first comprehensive longitudinal NVC evaluation during normal pregnancy reports significant but physiological microvascular variations throughout gestation, suggesting NVC as a safe and promising technique for further investigate and define patterns of microvascular changes also in pathological pregnancies.


Molecular characterization of bile salt hydrolase from Bifidobacterium animalis subsp. lactis Bi30.
The present work describes the identification, purification, and characterization of bile salt hydrolase (BSH) from Bifidobacterium animalis subsp. lactis. The enzyme was purified to electrophoretic homogeneity by hydrophobic chromatography, ion-exchange chromatography and ultrafiltration. SDS-PAGE analysis of putative BSH and gel filtration revealed that the analyzed protein is presumably a tetramer composed of four monomers each of about 35 kDa. The purified enzyme was analyzed by liquid chromatography coupled to LTQ FT ICR mass spectrometry and unambiguously identified as a bile salt hydrolase from B. animalis. The isoelectric point of the studied protein was estimated to be around pH 4.9. The pH optimum of the purified BSH is between 4.7 to 6.5, and the temperature optimum is around 50 degrees C. The BSH of B. animalis could deconjugate all tested bile salts, with clear preference for glycine-conjugated bile salts over taurine-conjugated forms. Genetic analysis of the bsh showed high similarity to the previously sequenced bsh gene from B. animalis and confirmed the usefulness of bile salt hydrolase as a genetic marker for B. animalis identification.


An ultra-high Q silicon compound cantilever resonator for Young's modulus measurements.
We describe the design of ultra-high Q mechanical cantilever resonators, fabricated from single-crystal silicon wafers. The mechanical resonance mode at f ≈ 8.5 kHz achieves a background damping of Q(-1) </~ 2 × 10(-8) at temperatures below 30 K, which is equal to that of a successful silicon torsional resonator with which the cantilever resonator shares several design elements. The new resonator can be used for accurate measurements of the Young's modulus and internal friction of thin films. It is compatible with both the mounting apparatus and measurement electronics of the torsional resonator, and the two resonators together can be used to provide a complete description of the elastic properties of isotropic thin films.


Characteristics and distribution of complex fractionated atrial electrograms and the dominant frequency during atrial fibrillation: relationship to the response and outcome of circumferential pulmonary vein isolation.
Although sites of complex fractionated electrograms (CFAEs) and dominant frequency (DF) are known to be critical for the maintenance of atrial fibrillation (AF), spatial distribution of CFAEs and DF and their impact on the outcome of AF ablation remain unclear. We created CFAE and DF maps of the left atrium (LA), right atrium, and pulmonary veins (PVs) with a NavX mapping system and simultaneously calculated the DF values with a Bard LabSystem Pro in 40 patients with AF (nonparoxysmal, n = 16). In 19 patients in whom circumferential PV isolation (CPVI) terminated AF, there was a high DF in the PVs (Bard-based DF value, 6.70 ± 1.01 Hz), low DF in the LA body (5.94 ± 0.75 Hz), and a significant PV-to-LA body DF gradient (0.76 ± 0.65 Hz), and the CFAEs were located mainly in the PV antrum. In the 21 patients not responding to CPVI, a high DF was located in both the PVs (7.04 ± 0.81 Hz) and LA body (6.75 ± 0.81 Hz), and therefore, the PV-to-LA body DF gradient was smaller than that in the CPVI responders (0.29 ± 0.52 Hz, P = 0.0160), and the CFAEs extended to the LA body. The higher DF in the LA body, nonparoxysmal AF, and longer AF duration remained as independent predictors of a post-ablation AF recurrence by using a multivariate analysis. A higher LA-DF value, smaller PV-to-LA DF gradient, and wider LA-CFAE distribution were noted more often in the nonresponders to CPVI than in the responders. This suggested the presence of an arrhythmogenic substrate in the LA beyond the PVs in patients whose AF persisted after CPVI, which was further associated with post-ablation AF recurrence.


Acute bronchial obstruction in sheep: histopathology and gland cytokine expression.
An ovine model of smoke inhalation and burn (S+B) injury models the pathophysiology of these injuries in humans. This study examines the degree of airway obstruction, associated histopathology, and bronchial gland cell expression of cytokines during the first 24 hours after S+B injury in sheep. Changes in the mean degree of obstruction were limited to the bronchial airways, showing significant increases in obstruction with time, P<.05. At 4 hours after injury, the obstructive material was predominantly mucus, with neutrophils clustered around and within gland acini. At 8 to 24 hours, bronchial obstruction was characterized by increased inflammatory cell accumulation. Immunohistochemical results showed that gland cells constitutively express and secrete interleukin (IL)-1beta, and that after injury there is an increase in the percentage of gland cells staining for IL-1alpha, IL-8, and tumor necrosis factor (TNF)-alpha, P<.05.


First report of Cercospora cf. malloti causing brown eyespot on strawberry in Brazil.
Strawberry (Fragaria x ananassa Duch.) is one of the most consumed fruits from the Rosaceae family in Brazil. It is cultivated on ~4,500 ha in all regions of the country, with a wide range of fungi causing leaf diseases. Strawberry leaves (cv. San Andreas) with symptoms of eyespot usually related to Mycosphaerella fragariae (Ramularia tulasnei) (Mazaro et al. 2006) were observed in April 2019, in a field located in Santa Maria, Rio Grande do Sul state, Brazil (29°43'19.6"S / 53°43'04.6"W). However, the symptoms slightly differed from Mycospherella blight as well as the fungi structures observed under an optical microscope. The symptomatic leaves had small circular spots, initially reddish-purple to dark brown color, which later increased in size. The center of the lesion was grayish pale, where dark brown conidiophores were found over it. The symptom occurred in 50% of the leaves of all plants during the crop season. To identify the causal agent, symptomatic leaves were collected and superficially disinfected with 70% ethanol for 30 s followed by 0.5% bleach for 30 s and then washed in sterile water. The leaves were placed in germination boxes with a moistened paper filter and incubated at 25 ºC and 12 h photoperiod for four days. After the formation of the fungal structures, the fungi was isolated in potato dextrose agar (PDA) medium and kept for 20 days at 25 ºC. Colonies were dark brown with pale pink aerial mycelium. The conidia were hyaline, single- to multi-septated with a filiform shape, straight or slightly curved. The fasciculate conidiophores on the stroma measured 20 to 70 x 2.5 to 3.5 µm. Conidia average measurement was 10.2 to 82.4 x 4. to 8.8 µm, similar to morphological characteristics of the genus Cercospora. The isolate named M6 was deposited in the Brazilian National System of Genetic Registration (SISGEN) no A57AE19. The DNA was extracted from a monosporic culture of M6 in PDA plates, which presented all typical aspects previously described. To confirm the identity, the PCR amplification was carried out for internal transcribed spacer (ITS) region ITS 1-5.8S - ITS 2 and Calmodulin region (CAL) genes (Acqua et al, 2011). The ITS (No. MZ713249) and CAL (No. MZ713254) sequences were deposited in the Genbank database. BLASTn analysis revealed the isolate was 99% similar to Cercospora cf. malloti which access codes in GenBank are KT 193689 and KT 193738 (Nguanhom et al. 2015). A pathogenicity test was performed to fulfill Koch's postulates. Strawberry plants with healthy leaves (60 days old), were inoculated by spraying a conidial suspension (1 × 105 conidia/ml). Control plants were sprayed with sterile water. The plants were incubated in a greenhouse at 28 ºC for 12 h photoperiod at 80% relative humidity under drip irrigation keeping at least 60% of field capacity. Six plants were used as replicates. Twenty days after inoculation, small circular reddish-purple spots were found, which later increased in size. Ten days later, the spots presented dark brown conidiophores over the center of the lesion. The symptoms described above were observed in all inoculated leaves. The control plants remained symptom-free. The pathogen was then re-isolated from the lesions and identified as C. cf. malloti based on the morphological aspects described above on PDA. In Thailand C. cf. malloti is reported on crops like Brassica alboglabra (cabbage) (Nguanhom et al. 2015), but not in strawberries. To our knowledge, this is the first report of C. cf. malloti pathogenic to strawberries in Brazil.


Nurses' attitudes toward patient and family education: implications for clinical nurse specialists.
This study had two purposes: (1) to examine nurses' attitudes toward patient and family education; and (2) to identify issues, barriers, and concerns related to patient and family education. A three-part survey was distributed to licensed nurses employed in a large healthcare system in Central Florida. Analysis of variance with posthoc multiple comparisons found differences in responses between different demographic groups. A model for meeting the patients' and families' education needs is proposed, using the various roles of the clinical nurse specialist to facilitate the process.


Biological invasion costs reveal insufficient proactive management worldwide.
The global increase in biological invasions is placing growing pressure on the management of ecological and economic systems. However, the effectiveness of current management expenditure is difficult to assess due to a lack of standardised measurement across spatial, taxonomic and temporal scales. Furthermore, there is no quantification of the spending difference between pre-invasion (e.g. prevention) and post-invasion (e.g. control) stages, although preventative measures are considered to be the most cost-effective. Here, we use a comprehensive database of invasive alien species economic costs (InvaCost) to synthesise and model the global management costs of biological invasions, in order to provide a better understanding of the stage at which these expenditures occur. Since 1960, reported management expenditures have totalled at least US$95.3 billion (in 2017 values), considering only highly reliable and actually observed costs - 12-times less than damage costs from invasions ($1130.6 billion). Pre-invasion management spending ($2.8 billion) was over 25-times lower than post-invasion expenditure ($72.7 billion). Management costs were heavily geographically skewed towards North America (54%) and Oceania (30%). The largest shares of expenditures were directed towards invasive alien invertebrates in terrestrial environments. Spending on invasive alien species management has grown by two orders of magnitude since 1960, reaching an estimated $4.2 billion per year globally (in 2017 values) in the 2010s, but remains 1-2 orders of magnitude lower than damages. National management spending increased with incurred damage costs, with management actions delayed on average by 11 years globally following damage reporting. These management delays on the global level have caused an additional invasion cost of approximately $1.2 trillion, compared to scenarios with immediate management. Our results indicate insufficient management - particularly pre-invasion - and urge better investment to prevent future invasions and to control established alien species. Recommendations to improve reported management cost comprehensiveness, resolution and terminology are also made.


Percentage of Age-Predicted Cardiorespiratory Fitness and Risk of Incident Hypertension: A PROSPECTIVE COHORT STUDY.
There are inverse and independent associations between cardiorespiratory fitness (CRF) and several adverse cardiometabolic outcomes including hypertension (HTN). The prospective relationship between percentage of age-predicted CRF and risk of HTN has not been previously investigated. We aimed to assess the association of percentage of age-predicted CRF with incident HTN in a long-term prospective cohort study. Cardiorespiratory fitness, as measured by peak oxygen uptake, was assessed using the gold standard respiratory gas exchange analyzer in 1602 men who underwent cardiopulmonary exercise testing. The age-predicted CRF estimated from a regression equation for age was converted to percentage of age-predicted CRF ([achieved CRF/age-predicted CRF] × 100). The HR with 95% CI were estimated for HTN. During a median follow-up of 26.8 yr, 308 HTN cases were recorded. There was a linear relationship between age-predicted CRF and incident HTN ( P value for nonlinearity = .68). A 1-SD increase in percentage of age-predicted CRF was associated with a decreased risk of HTN in analysis adjusted for established risk factors (HR = 0.79: 95% CI, 0.69-0.90), which remained consistent on further adjustment for several other potential confounders including alcohol consumption, physical activity, socioeconomic status, and inflammation (HR = 0.83: 95% CI, 0.72-0.95). The corresponding adjusted HRs were 0.55 (95% CI, 0.38-0.80) and 0.63 (95% CI, 0.43-0.91), respectively, when comparing extreme quartiles of percentage of age-predicted CRF levels. Percentage of age-predicted CRF is linearly, inversely, and independently associated with risk of incident HTN, and it is comparable to absolute CRF as a risk indicator for HTN in a general population of middle-aged men.


Early Identification of Fungal and Mycobacterium Infections in Pulmonary Granulomas Using Metagenomic Next-Generation Sequencing on Formalin fixation and paraffin embedding tissue.
The purpose of the study is to assess the etiology detection ability of Metagenomic Next-Generation Sequencing (mNGS) on formalin fixation and paraffin embedding (FFPE) tissue from postoperative biopsy specimens. We prospectively enrolled specimens from patients undergone surgery biopsy due to undefinite diagnosis and pathologically indicated granulomatous lesions. FFPE tissues were tested by mNGS and histopathology. The etiology detection rate of mNGS was calculated and compared with histopathology.. Among the 69 cases eventually included, 41 (59.42%) were diagnosed with infectious granuloma. The overall fungi and mycobacteria etiology detection rates of mNGS in granuloma lesions was 87.80% (36/41). The mNGS increased the detection rate by 68.29% (28/41) compared with histopathology, the difference was statistically significant (χ2 = 28.97, P = 0.00). The detection rates of mNGS in fungal infections (12/12,100%) and in mycobacterium infections (22/27, 81.48%) were significant higher than those of histopathology (8/12, 66.67% and 0/27, 0.00%; both P = 0.00). Two (2/2.100%) cases of co-infection were detected at one time by mNGS. All mNGS-based clinical decisions were made within 2 days. The mNGS could accurately and quickly detect fungi and mycobacteria in FFPE specimens from postoperative granuloma specimens and identify the pathogens to the species level. China Clinical Trial Registry ChiCTR2000035464.


An assay of Bordetella pertussis adhesion to tissue-culture cells.
The ability of Bordetella pertussis to bind to cell surfaces was determined with a simple, accurate, reproducible assay measuring the adhesion of radiolabelled bacteria to monolayers of HeLa cells. The rate of adhesion was approximately linear with time for at least 1 h. Viable and radioactivity counts of bound bacteria correlated well. Bacteria grown in the avirulent C-mode were markedly less adhesive than virulent X-mode cells. Reductions in the level of attachment after treatment of bacteria with preparations of specific immunoglobulin suggest that adhesion of B. pertussis depends upon specific mechanisms involving filamentous haemagglutinin and the agglutinogens.


Population genomic analysis of the rice blast fungus reveals specific events associated with expansion of three main clades.
We examined the genomes of 100 isolates of Magnaporthe oryzae (Pyricularia oryzae), the causal agent of rice blast disease. We grouped current field populations of M. oryzae into three major globally distributed groups. A genetically diverse group, clade 1, which may represent a group of closely related lineages, contains isolates of both mating types. Two well-separated clades, clades 2 and 3, appear to have arisen as clonal lineages distinct from the genetically diverse clade. Examination of genes involved in mating pathways identified clade-specific diversification of several genes with orthologs involved in mating behavior in other fungi. All isolates within each clonal lineage are of the same mating type. Clade 2 is distinguished by a unique deletion allele of a gene encoding a small cysteine-rich protein that we determined to be a virulence factor. Clade 3 isolates have a small deletion within the MFA2 pheromone precursor gene, and this allele is shared with an unusual group of isolates we placed within clade 1 that contain AVR1-CO39 alleles. These markers could be used for rapid screening of isolates and suggest specific events in evolution that shaped these populations. Our findings are consistent with the view that M. oryzae populations in Asia generate diversity through recombination and may have served as the source of the clades 2 and 3 isolates that comprise a large fraction of the global population.


Asymptomatic hypoxia in COVID-19 is associated with poor outcome.
Describe and evaluate the outcome of a coronavirus disease-2019 (COVID-19) patient without shortness of breath. We retrospectively collected data from COVID-19 patients diagnosed and cared for in Marseille, France. We selected data from patients who at admission, had a low dose CT scanner, dyspnea status, and oxygen saturation available. Blood gas was analyzed in a sample subset of patients. Among 1712 patients with COVID-19, we report that 1107 (64.7%) do not complain of shortness of breath at admission. The low-dose computed tomography (LDCT) scan showed signs compatible with pneumonia in 757/1,107 (68.4%) of patients without dyspnea. In a subset of patients who had underwent at least one blood gas analysis (n = 161) and presented without dyspnea at admission, 28.1% (27/96) presented with a hypoxemia/hypocapnia syndrome. Asymptomatic hypoxia was associated with a very poor outcome (33.3% were transferred to the ICU and 25.9% died). The absence of shortness of breath in an old patient with comorbidity merit medical attention and should not be considered as a good sign of well-being. The poor prognosis of asymptomatic hypoxia, highlight the severity of this mild clinical presentation. In these patients, pulse oximetry is an important mean to predict the outcome along with news score and LDCT scanner.


Comparison of the oral effectiveness of etilefrine pivalate and etilefrine in a long-term trial in man.
The influence of equimolar oral doses of etilefrine pivalate (K 30 052, Ep) and etilefrine on blood pressure of patients suffering from orthostatic dysregulation was compared. Both drugs augmented the systolic blood pressure in both, the upright and horizontal body posture. Diastolic blood pressure was not influenced. The resulting increase in blood pressure amplitude points to an augmentation of cardiac stroke volume. Ep in equimolar doses was nearly twice as active as etilefrine. This result is in line with the hypothesis that acylation of etilefrine with pivalinic acid inhibits the first-pass inactivation nearly completely.


Temporal Modulation of Differential Alternative Splicing in HaCaT Human Keratinocyte Cell Line Chronically Exposed to Arsenic for up to 28 Wk.
Chronic arsenic exposure via drinking water is associated with an increased risk of developing cancer and noncancer chronic diseases. Pre-mRNAs are often subject to alternative splicing, generating mRNA isoforms encoding functionally distinct protein isoforms. The resulting imbalance in isoform species can result in pathogenic changes in critical signaling pathways. Alternative splicing as a mechanism of arsenic-induced toxicity and carcinogenicity is understudied. This study aimed to accurately profile differential alternative splicing events in human keratinocytes induced by chronic arsenic exposure that might play a role in carcinogenesis. Independent quadruplicate cultures of immortalized human keratinocytes (HaCaT) were maintained continuously for 28 wk with 0 or 100 nM sodium arsenite. RNA-sequencing (RNA-Seq) was performed with poly(A) RNA isolated from cells harvested at 7, 19, and 28 wk with subsequent replicate multivariate analysis of transcript splicing (rMATS) analysis to detect and quantify differential alternative splicing events. Reverse transcriptase-polymerase chain reaction (RT-PCR) for selected alternative splicing events was performed to validate RNA-Seq predictions. Functional enrichment was performed by gene ontology (GO) analysis of the differential alternative splicing event data set at each time point. At least 600 differential alternative splicing events were detected at each time point tested, comprising all the five main types of alternative splicing and occurring in both open reading frames (ORFs) and untranslated regions (UTRs). Based on functional relevance ELK4, SHC1, and XRRA1 were selected for validation of predicted alternative splicing events at 7 wk by RT-PCR. Densitometric analysis of RT-PCR data corroborated the rMATS predicted alternative splicing for all three events. Protein expression validation of the selected alternative splicing events was challenging given that very few isoform-specific antibodies are available. GO analysis demonstrated that the enriched terms in differential alternatively spliced mRNAs changed dynamically with the time of exposure. Notably, RNA metabolism and splicing regulation pathways were enriched at the 7-wk time point, when the greatest number of differentially alternatively spliced mRNAs are detected. Our preliminary proteomic analysis demonstrated that the expression of the canonical isoforms of the splice regulators DDX42, RMB25, and SRRM2 were induced upon chronic arsenic exposure, corroborating the splicing predictions. These results using cultures of HaCaT cells suggest that arsenic exposure disrupted an alternative splice factor network and induced time-dependent genome-wide differential alternative splicing that likely contributed to the changing proteomic landscape in arsenic-induced carcinogenesis. However, significant challenges remain in corroborating alternative splicing data at the proteomic level. https://doi.org/10.1289/EHP9676.


A kinetic Ising model study of dynamical correlations in confined fluids: emergence of both fast and slow time scales.
Experiments and computer simulation studies have revealed existence of rich dynamics in the orientational relaxation of molecules in confined systems such as water in reverse micelles, cyclodextrin cavities, and nanotubes. Here we introduce a novel finite length one dimensional Ising model to investigate the propagation and the annihilation of dynamical correlations in finite systems and to understand the intriguing shortening of the orientational relaxation time that has been reported for small sized reverse micelles. In our finite sized model, the two spins at the two end cells are oriented in the opposite directions to mimic the effects of surface that in real system fixes water orientation in the opposite directions. This produces opposite polarizations to propagate inside from the surface and to produce bulklike condition at the center. This model can be solved analytically for short chains. For long chains, we solve the model numerically with Glauber spin flip dynamics (and also with Metropolis single-spin flip Monte Carlo algorithm). We show that model nicely reproduces many of the features observed in experiments. Due to the destructive interference among correlations that propagate from the surface to the core, one of the rotational relaxation time components decays faster than the bulk. In general, the relaxation of spins is nonexponential due to the interplay between various interactions. In the limit of strong coupling between the spins or in the limit of low temperature, the nature of relaxation of the spins undergoes a qualitative change with the emergence of a homogeneous dynamics where decay is predominantly exponential, again in agreement with experiments.


Formation of disinfection by-products after pre-oxidation with chlorine dioxide or ferrate.
The effect of pre-oxidation with chlorine dioxide (ClO2) or ferrate (Fe(VI)) on the formation of disinfection by-products (DBPs) during chlorination or chloramination was tested with natural waters from 12 sources (9 surface waters, 1 groundwater, and 2 wastewater effluents). DBPs investigated included trihalomethanes (THM), chloral hydrate (CH), haloketones (HK), haloacetonitriles (HAN) and trichloronitromethane (TCNM), chlorite and chlorate. Chlorite and chlorate were found in the ClO2-treated waters. Application of 1 mg/L ClO2 ahead of chlorination reduced the formation potential for THM by up to 45% and the formation of HK, HAN and TCNM in most of the samples. The CH formation results were mixed. The formation of CH and HK was enhanced with low doses of Fe(VI) (1 mg/L as Fe), but was greatly reduced at higher doses (20 mg/L Fe). Fe(VI) reduced the formation of THM, HAN and TCNM in most of the samples. Reduced potential for the formation of NDMA was observed in most of the samples after both ClO2 and Fe(VI) pre-oxidation.


The tetramerization region of the retinoid X receptor is important for transcriptional activation by the receptor.
The retinoid X receptor (RXR), a member of the superfamily of hormone nuclear receptors, is a ligand-inducible transcription factor that is activated by the vitamin A derivative 9-cis-retinoic acid. We previously showed that RXR self-associates into tetramers with a high affinity and that ligand binding induces rapid dissociation of receptor tetramers to smaller species. Here, the RXR region that is responsible for mediating tetramer formation is identified. It is shown that this interface, which we term the "tetramerization domain," critically contains two consecutive phenylalanine residues located at the C-terminal region of the receptor. Mutation of these residues is sufficient to disrupt RXR tetramers without affecting the overall fold of the protein or interfering with ligand binding, dimer formation, or DNA binding by the receptor. Nevertheless, the tetramer-impaired mutant was found to be transcriptionally defective. The newly characterized tetramerization domain and the previously identified main dimerization interface of RXR act autonomously to affect separate intersubunit interactions that, overall, lead to formation of tetramers. Protein-protein interactions mediated by the tetramerization domain, but not those that involve the dimerization interface, are disrupted following ligand binding by RXR. Overall, these data attest to the specificity of the interaction and implicate the tetramerization interface in playing a direct role in regulating transcriptional activation by RXR.


The relationship between expectations and satisfaction: a qualitative study of patients' experiences of surgery for gynaecological cancer.
It is important that a patient perspective is introduced to the identification and measurement of the outcomes of health care. The aim of this study was to use qualitative methods to examine the presence or absence of expectations prior to the experience of health care and the relationship between expectations, satisfaction and dissatisfaction in a group of women undergoing surgery in a large teaching hospital. Nineteen women with a diagnosis of gynaecological cancer were interviewed on two occasions, before and after surgery. A thematic analysis was undertaken. The results suggest that there is not a clear relationship between expectations and satisfaction. Women had different levels of expectations about different types of care and different aspects of care. Unfulfilled expectations did not lead to less satisfaction. The women were able to express satisfaction either with the care overall or with specific aspects of care, as well as being able to distinguish aspects of care with which they were dissatisfied.


Rat McA-RH7777 cells efficiently assemble rat apolipoprotein B-48 or larger fragments into VLDL but not human apolipoprotein B of any size.
Studies of truncated apoB peptides in human subjects with familial hypobetalipoproteinemia, as well as of puromycin-generated spectra of nascent apoB peptides in rat and hamster liver, suggest that a minimum size is required for N-terminal fragments of apoB to be efficiently assembled into full-sized VLDL. We report here results of experiments undertaken to examine this phenomenon in greater detail by expressing individual carboxyl-truncated human apoB constructs in McArdle cells. Thus, apoB-29, -32, -37, -42, -47, -53, -70 and full length apoB-100 were transiently expressed in rat McA-RH7777 hepatoma cells, or human apoB-31 and apoB-53 were stably expressed in the same cells, and the secreted VLDL particles were characterized by kinetic gradient ultracentrifugal flotation. Calibration with rat plasma VLDL subfractions showed that about 90 and 50%, respectively, of lipoprotein particles containing endogenous rat B-100 and B-48 floated between fractions 2;-8 of the 11-fraction gradient. This corresponds to the normal VLDL diameter range of about 47 to 28 nm, with the remaining half of rat B-48 recovered as HDL particles in the 1.1 g/ml range. In contrast, regardless of their size, only 2;-5% of any of the truncated human apoB peptides expressed in these cells was recovered in the VLDL region of the gradient. The remaining 95+% of the lipoproteins were found as high density particles; as previously found in other systems the densities of the latter were inversely related to their peptide chain-length. Furthermore, transiently expressed full-length human apoB-100 was inefficiently secreted as VLDL by these cells, with the remainder appearing as LDL-sized particles. Thus, although we showed that McA-RH7777 cells secreted endogenous rat apoB as normal-sized VLDL, we found them unsuitable for our original purpose of using human apoB fragments to further define effects of apoB size on VLDL assembly. These cells appeared unable to efficiently use any size of human apoB for that process. Pulse-labeled untransfected McA-RH7777 cells chased in the presence of puromycin did, however, show a sharp decline in VLDL assembly efficiency for endogenous nascent rat apoB peptides shorter than B-48, similar to that originally found in normal rat liver.


Descemet's membrane detachment managed with perfluro-n-octane liquid.
We report the case of a 68-year-old male who developed Descemet's membrane detachment after temporal clear corneal phacoemulsification which did not settle with air or viscoelastic injection. The Descemet's membrane was successfully reattached with restoration of 20/50 vision with the help of perfluro-n-octane liquid. To our knowledge, this is the first such case to be reported.


Effects of high glucose and thiamine on the balance between matrix metalloproteinases and their tissue inhibitors in vascular cells.
Pericyte survival in diabetic retinopathy depends also on interactions with extracellular matrix (ECM) proteins, which are degraded by matrix metalloproteinases (MMP). Elevated glucose influences ECM turnover, through expression of MMP and their tissue inhibitors, TIMP. We reported on reduced pericyte adhesion to high glucose-conditioned ECM and correction by thiamine. We aimed at verifying the effects of thiamine and benfotiamine on MMP-2, MMP-9 and TIMP expression and activity in human vascular cells with high glucose. In HRP, MMP-2 activity, though not expression, increased with high glucose and decreased with thiamine and benfotiamine; TIMP-1 expression increased with high glucose plus thiamine and benfotiamine; MMP-9 was not expressed. In EC, MMP-9 and MMP-2 expression and activity increased with high glucose, but thiamine and benfotiamine had no effects; TIMP-1 expression was unchanged. Neither glucose nor thiamine modified TIMP-2 and TIMP-3 expression. TIMP-1 concentrations did not change in either HRP or EC. High glucose imbalances MMP/TIMP regulation, leading to increased ECM turnover. Thiamine and benfotiamine correct the increase in MMP-2 activity due to high glucose in HRP, while increasing TIMP-1.


Reduced apoptosis and ameliorated listeriosis in TRAIL-null mice.
Listeriosis is an infectious disease caused by the bacterium Listeria monocytogenes. Although it is well recognized that apoptosis plays a critical role in the pathogenesis of the disease, the molecular mechanisms of cell death in listeriosis remain to be established. We report in this study that mice deficient in TRAIL were partially resistant to primary listeriosis, and blocking TRAIL with a soluble death receptor 5 markedly ameliorated the disease. The numbers of Listeria in the liver and spleen of TRAIL+/+ mice were 10-100 times greater than those in TRAIL-/- mice following primary Listeria infection. This was accompanied by a significant increase in the survival rate of TRAIL-/- mice. Lymphoid and myeloid cell death was significantly inhibited in TRAIL-/- mice, which led to marked enlargement of the spleen. These results establish a critical role for TRAIL in apoptosis during listeriosis.


Biventricular pacing worsened dyssynchrony in heart failure patient with right-bundle branch block.
The use of a biventricular pacing system for patients with complete right-bundle branch block (CRBBB) is still controversial. Although cardiac resynchronization therapy-defibrillator (CRT-D) was implanted in a heart failure patient with CRBBB, dyssynchrony worsened and stroke volume decreased, and this patient was re-admitted due to exacerbated heart failure. Therefore, evaluation of dyssynchrony and cardiac function after implantation of a biventricular pacing system may be needed in patients with atypical indications for CRT.


Capturing three-dimensional in vivo lumbar intervertebral joint kinematics using dynamic stereo-X-ray imaging.
Availability of accurate three-dimensional (3D) kinematics of lumbar vertebrae is necessary to understand normal and pathological biomechanics of the lumbar spine. Due to the technical challenges of imaging the lumbar spine motion in vivo, it has been difficult to obtain comprehensive, 3D lumbar kinematics during dynamic functional tasks. The present study demonstrates a recently developed technique to acquire true 3D lumbar vertebral kinematics, in vivo, during a functional load-lifting task. The technique uses a high-speed dynamic stereo-radiography (DSX) system coupled with a volumetric model-based bone tracking procedure. Eight asymptomatic male participants performed weight-lifting tasks, while dynamic X-ray images of their lumbar spines were acquired at 30 fps. A custom-designed radiation attenuator reduced the radiation white-out effect and enhanced the image quality. High resolution CT scans of participants' lumbar spines were obtained to create 3D bone models, which were used to track the X-ray images via a volumetric bone tracking procedure. Continuous 3D intervertebral kinematics from the second lumbar vertebra (L2) to the sacrum (S1) were derived. Results revealed motions occurring simultaneously in all the segments. Differences in contributions to overall lumbar motion from individual segments, particularly L2-L3, L3-L4, and L4-L5, were not statistically significant. However, a reduced contribution from the L5-S1 segment was observed. Segmental extension was nominally linear in the middle range (20%-80%) of motion during the lifting task, but exhibited nonlinear behavior at the beginning and end of the motion. L5-S1 extension exhibited the greatest nonlinearity and variability across participants. Substantial AP translations occurred in all segments (5.0 ± 0.3 mm) and exhibited more scatter and deviation from a nominally linear path compared to segmental extension. Maximum out-of-plane rotations (<1.91 deg) and translations (<0.94 mm) were small compared to the dominant motion in the sagittal plane. The demonstrated success in capturing continuous 3D in vivo lumbar intervertebral kinematics during functional tasks affords the possibility to create a baseline data set for evaluating the lumbar spinal function. The technique can be used to address the gaps in knowledge of lumbar kinematics, to improve the accuracy of the kinematic input into biomechanical models, and to support development of new disk replacement designs more closely replicating the natural lumbar biomechanics.


Reduced differentiation potential of primary MyoD-/- myogenic cells derived from adult skeletal muscle.
To gain insight into the regeneration deficit of MyoD-/- muscle, we investigated the growth and differentiation of cultured MyoD-/- myogenic cells. Primary MyoD-/- myogenic cells exhibited a stellate morphology distinct from the compact morphology of wild-type myoblasts, and expressed c-met, a receptor tyrosine kinase expressed in satellite cells. However, MyoD-/- myogenic cells did not express desmin, an intermediate filament protein typically expressed in cultured myoblasts in vitro and myogenic precursor cells in vivo. Northern analysis indicated that proliferating MyoD-/- myogenic cells expressed fourfold higher levels of Myf-5 and sixfold higher levels of PEA3, an ETS-domain transcription factor expressed in newly activated satellite cells. Under conditions that normally induce differentiation, MyoD-/- cells continued to proliferate and with delayed kinetics yielded reduced numbers of predominantly mononuclear myocytes. Northern analysis revealed delayed induction of myogenin, MRF4, and other differentiation-specific markers although p21 was upregulated normally. Expression of M-cadherin mRNA was severely decreased whereas expression of IGF-1 was markedly increased in MyoD-/- myogenic cells. Mixing of lacZ-labeled MyoD-/- cells and wild-type myoblasts revealed a strict autonomy in differentiation potential. Transfection of a MyoD-expression cassette restored cytomorphology and rescued the differentiation deficit. We interpret these data to suggest that MyoD-/- myogenic cells represent an intermediate stage between a quiescent satellite cell and a myogenic precursor cell.


68Ga-DOTANOC Focal Pulmonary Activity With No Corresponding CT Abnormality.
A 28-year-old woman with medullary thyroid cancer treated with total thyroidectomy and bilateral central and right lateral neck nodal resection postoperatively had increasing calcitonin and carcinoembryonic antigen values. A Ga-DOTANOC PET/CT study performed showed tracer-avid disease in the neck, mediastinal, and hilar lymph nodes. However, there were 3 tracer-avid foci in the lungs bilaterally, suggestive of lung metastases but with no corresponding abnormalities on low-dose CT. A negative contrast-enhanced CT of the lungs along with clinical and radiological follow-up confirmed that the lung activity was artifactual, demonstrating the possibility of Ga-DOTANOC-avid iatrogenic pulmonary microembolism.


TVAR: assessing tissue-specific functional effects of non-coding variants with deep learning.
Analysis of whole-genome sequencing (WGS) for genetics is still a challenge due to the lack of accurate functional annotation of non-coding variants, especially the rare ones. As eQTLs have been extensively implicated in the genetics of human diseases, we hypothesize that rare non-coding variants discovered in WGS play a regulatory role in predisposing disease risk. With thousands of tissue- and cell-type-specific epigenomic features, we propose TVAR. This multi-label learning-based deep neural network predicts the functionality of non-coding variants in the genome based on eQTLs across 49 human tissues in the GTEx project. TVAR learns the relationships between high-dimensional epigenomics and eQTLs across tissues, taking the correlation among tissues into account to understand shared and tissue-specific eQTL effects. As a result, TVAR outputs tissue-specific annotations, with an average AUROC of 0.77 across these tissues. We evaluate TVAR's performance on four complex diseases (coronary artery disease, breast cancer, Type 2 diabetes and Schizophrenia), using TVAR's tissue-specific annotations, and observe its superior performance in predicting functional variants for both common and rare variants, compared with five existing state-of-the-art tools. We further evaluate TVAR's G-score, a scoring scheme across all tissues, on ClinVar, fine-mapped GWAS loci, Massive Parallel Reporter Assay (MPRA) validated variants and observe the consistently better performance of TVAR compared with other competing tools. The TVAR source code and its scores on the ClinVar catalog, fine mapped GWAS Loci, high confidence eQTLs from GTEx dataset, and MPRA validated functional variants are available at https://github.com/haiyang1986/TVAR. Supplementary data are available at Bioinformatics online.


High-buffering capacity, hydrolytically stable, low-pI isoelectric membranes for isoelectric trapping separations.
Hydrolytically stable, low-pI isoelectric membranes have been synthesized from low-pI ampholytic components, poly(vinyl alcohol), and a bifunctional cross-linker, glycerol-1,3-diglycidyl ether. The low-pI ampholytic components used contain one amino group and at least two weakly acidic functional groups. The acidic functional groups are selected such that the pI value of the ampholytic component is determined by the pK(a) values of the acidic functional groups. When the concentration of the ampholytic component incorporated into the membrane is higher than a required minimum value, the pI of the membrane becomes independent of variations in the actual incorporation rate of the ampholytic compound. The new, low-pI isoelectric membranes have been successfully used as anodic membranes in isoelectric trapping separations with pH < 1.5 anolytes and replaced the hydrolytically less stable polyacrylamide-based isoelectric membranes. The new low-pI isoelectric membranes have excellent mechanical stability, low electric resistance, good buffering capacity, and long life time, even when used with as much as 50 W power and current densities as high as 33 mA/cm(2) during the isoelectric trapping separations.


Revised NIOSH lifting equation: a critical evaluation.
The revised NIOSH lifting equation (RNLE) aims to manage lifting-related lower back pain (LBP), by determining safe load limits. Many researchers have studied the multiplier development criteria, the universal applicability of the equation and its ability to identify an increased risk of LBP in lifting tasks. Although a number of strengths of the equation have been highlighted, many limitations have also been identified. The need for new multipliers, such as worker and environmental characteristics, was highlighted in order to make the equation more adaptable. The RNLE was designed to protect 75% of female workers and is therefore inherently conservative. Additionally, as all multipliers have values less than or equal to 1, the recommended weight limits (RWLs) can be further reduced. Thus, new multipliers may be defined, by combining two or more existing multipliers, to make the RWLs more realistic.


Attenuation of renomedullary phospholipase C isozyme, PLC-delta 1, in spontaneously hypertensive rats.
The distributional patterns of PLC isozymes within the kidney were investigated using spontaneously hypertensive rats (SHRs) and normotensive Wistar-Kyoto (WKY) rats at 4 and 12 weeks of age. PLC-beta 1, PLC-beta 3 and PLC-delta 1 quantified by Western blot analysis, were present in the highest concentrations in the inner medulla of rats at both 4 and 12 weeks of age. On the other hand, PLC-beta 4, PLC-gamma 1 and PLC-gamma 2 were distributed almost equally among the regions for the rats of both ages. When compared with WKY rats at 12 weeks of age, the amounts of PLC-beta 1, PLC-beta 3, PLC-gamma 1, PLC-gamma 2, and PLC-delta 1 in the inner medulla of SHRs were significantly lower, and the amount of PLC-delta 1 in the inner stripe of the outer medulla was also significantly lower. Even at the prehypertensive stage at 4 weeks of age, the inner medullary concentration of PLC-delta 1 was significantly lower in SHRs than WKY rats. These results suggest that PLC-delta 1 would play an important role in the development of hypertension.


Intraabdominal Mycobacterium tuberculosis vs Mycobacterium avium-intracellulare infections in patients with AIDS: distinction based on CT findings.
Abdominal CT scans of 71 patients with AIDS who had proved disseminated infection due to Mycobacterium tuberculosis (27 patients) or Mycobacterium avium-intracellulare (44 patients) were reviewed retrospectively to identify radiologic features that can be used to distinguish the two infections. CT findings in patients with disseminated M. tuberculosis included focal lesions in the liver (11%), spleen (30%), kidneys (19%), pancreas (7%), and gastrointestinal tract (15%) and lymph nodes with central or diffuse low attenuation (93%). CT findings in patients with disseminated M. avium-intracellulare included marked hepatomegaly (20%); marked splenomegaly (14%); focal lesions in the liver (9%), spleen (7%), and kidneys (2%); diffuse jejunal wall thickening (18%); lymph nodes with central low attenuation (14%); and enlarged lymph nodes exclusively of homogeneous soft-tissue density (55%). The presence of focal visceral lesions and low-attenuation lymph nodes suggests disseminated M. tuberculosis, whereas marked hepatic and splenic enlargement, diffuse jejunal wall thickening, and enlarged soft-tissue-density lymph nodes suggest disseminated M. avium-intracellulare. Recognition of these CT features can lead to a tentative diagnosis so that appropriate therapy can be instituted before the results of mycobacterial cultures become available.


Phytosterols and inulin-enriched soymilk increases glucagon-like peptide-1 secretion in healthy men: double-blind randomized controlled trial, subgroup study.
The study aimed to determine the effect of phytosterols and inulin on plasma glucose, insulin, and GLP-1 levels among healthy men after consuming phytosterols and inulin-enriched soymilk for 8 weeks. A total of 26 men at least 20 years old were randomly assigned into the 2 g/day of phytosterols and 10 g/day of inulin-enriched soymilk (intervention) group or into the standard soymilk (control) group. In the intervention group, the area under the curve of Glucagon-like peptide-1 secretion increased significantly, compared to its baseline (p = 0.003). The area under the curve of insulin secretion also increased but it did not meet statistical significance (p = 0.118). The area under the curves of plasma glucose were similar between pre- and post-test (p = 0.348). In the control group, none of the primary results significantly changed compared to their baseline levels. Trial registration Thai Clinical Trial Registry: TCTR20160319001 date: March 19, 2016, retrospectively registered.


Merkel cell carcinoma of the skin: imaging and clinical features in 93 cases.
Clinical, pathological and imaging data for 93 patients with Merkel cell carcinoma were reviewed. Median survival duration from the date of diagnosis was estimated using the Kaplan-Meier product-limit method. Survival durations of patients with no metastastic disease, with regional lymph node metastasis only and with distant metastases were compared using the Cox-Mantel log-rank test. The median age for patients with Merkel cell carcinoma was 70 years (range 35-91 years). The tumour occurred in 71 men and 22 women. The most common primary site was the head and neck region (64 patients), followed by the extremities (21 patients) and the trunk (10 patients). Median survival duration for the whole group was 33 months (range 2-140 months). Imaging studies of the primary sites were available for 10 patients and of metastatic sites for seven patients. Sonograms of the primary skin lesions showed single or multicentric hypoechoic nodules arising from the dermis and growing into the subcutaneous tissues. CT scans showed single or multiple minimally enhancing soft tissue nodules, in two instances associated with lytic bone erosion. The metastatic lesions were moderately hypoechoic on sonography and target-shaped with or without ring-enhancement on CT. Imaging studies have a limited role in diagnosis of the clinically evident primary skin lesions, but may be helpful in the assessment of the depth of the invasion and are crucial in the evaluation of regional and distant metastatic disease. If regional adenopathy is found then CT of the chest, abdomen and pelvis may be in order.


[Detection of IFN Response of Non-Specific Effects on RNAi.].
The RNA interference (RNAi) approach is an efficient and widely used method for silencing specific genes and exploring their functions. However, numerous studies have highlighted the non-specific effects of RNAi experiments and the IFN response is one of the most common non-specific effects. In this research, we will study how to detect the IFN response in RNAi experiments. Five different FLJ20420 siRNAs was transfected into human lung adenocarinoma cell line A549 using LipofectamineTM2000. The alternation of the interferon stimulated genes (ISGs) expression profile was analysed by real-time PCR and microarray assay. (1) Different FLJ20420 siRNA had different effect on silencing FLJ20420 mRNA expression in A549 cells, and FLJsiRNA-1 and -4 had the better effects to knockdown the expression of FLJ20420 (80% and 90% down-regulation, respectively); (2) In analyzed 16 major ISGs, the expression of 14 of these genes was significantly increased in A549-FLJsiRNA-1 cells (P <0.05), while only 2 of these genes had a significant change in A549-FLJ-siRNA-4 cells. These results indicated an IFN response existed in A549-FLJ-siRNA-1 cells. (3) Microarray assay indicated that the expression of 51 ISGs were increased over 2 times in A549-FLJ-siRNA-1 cells, while only the expression of 6 ISGs were over 2-time in A549-FLJ-siRNA-4 cells. These results confirmed that there was an IFN response of non-specific effects of RNAi in A549 cells transfected with FLJ-siRNA-1. In RNAi experiments, siRNA can not only silence the target genes, but also may activate IFN response. The analysis of the expression change of the major ISGs by real-time PCR is a useful approach to explore the IFN response in RNAi experiments.


Hypoplastic left heart syndrome associated with congenital right-sided diaphragmatic hernia and omphalocele.
Congenital diaphragmatic hernia (CDH) is associated with a variety of cardiac anomalies. However, its association with hypoplastic left heart syndrome (HLHS) is rare. We treated a female newborn with CDH, HLHS, and omphalocele. The operation for omphalocele and the diaphragmatic defect was successful, although the patient died of cardiac failure after Norwood's operation for HLHS. To our knowledge, this is the first reported case with a combination of these three major anomalies: CDH, HLHS, and omphalocele.


The role of ultrasound in the surgical management of patients diagnosed with ductal carcinoma in situ of the breast.
The purpose of this study was to determine the usefulness of ultrasound (US) as an adjunct to mammography (MMG) in the surgical treatment planning for patients with ductal carcinoma in situ (DCIS) of the breast. A total of 119 patients diagnosed with DCIS, who were treated between 1999 and 2002, were identified from the institutional database. US and MMG size of suspicious abnormalities, pathologic tumor size, and findings of axillary US and surgical axillary evaluation were analyzed. The median size difference of US versus pathologic tumor size and MMG versus pathologic tumor size was 1.0 and 2.0 cm, respectively. Correlation coefficients for US versus pathologic tumor size and MMG versus pathologic tumor size were 0.53 and -0.09, respectively. The negative and positive predictive values of axillary US to predict nodal disease were 93% and 27%, respectively. US evaluation appears to be a useful tool in conjunction with MMG in determining the extent of disease in patients diagnosed with DCIS. However, the low positive predictive value of US for evaluating metastases to axillary lymph nodes does not justify the routine use of this modality for axillary evaluation in patients with DCIS. Therefore the use of US for patients diagnosed with DCIS needs further investigation.


Urethral reconstruction using an autologous bladder mucosal graft.
Since 1988 twelve children underwent urethral reconstruction by using an autologous bladder mucosal graft. In 10 of these previous surgery for hypospadias or other penile anomalies had failed. Bladder mucosal graft was used in 2 cases for primary urethral reconstruction. Our results in using this method, mostly in hypospadias cripples, are satisfying, so that we are encouraged to use the autologous bladder mucosal graft for primary urethral reconstruction in selected cases with severe forms of hypospadias as well.


Shaken Baby Syndrome: Magnetic Resonance Imaging Features in Abusive Head Trauma.
In the context of child abuse spectrum, abusive head trauma (AHT) represents the leading cause of fatal head injuries in children less than 2 years of age. Immature brain is characterized by high water content, partially myelinated neurons, and prominent subarachnoid space, thus being susceptible of devastating damage as consequence of acceleration-deceleration and rotational forces developed by violent shaking mechanism. Diagnosis of AHT is not straightforward and represents a medical, forensic, and social challenge, based on a multidisciplinary approach. Beside a detailed anamnesis, neuroimaging is essential to identify signs suggestive of AHT, often in absence of external detectable lesions. Magnetic resonance imaging (MRI) represents the radiation-free modality of choice to investigate the most typical findings in AHT, such as subdural hematoma, retinal hemorrhage, and hypoxic-ischemic damage and it also allows to detect more subtle signs as parenchymal lacerations, cranio-cervical junction, and spinal injuries. This paper is intended to review the main MRI findings of AHT in the central nervous system of infants, with a specific focus on both hemorrhagic and non-hemorrhagic injuries caused by the pathological mechanisms of shaking. Furthermore, this review provides a brief overview about the most appropriate and feasible MRI protocol to help neuroradiologists identifying AHT in clinical practice.