Molar pregnancy, childhood cancer and genomic imprinting - is there a link?
The United Kingdom Childhood Cancer Study (UKCCS) is a national multi-centre case-control study that was designed to evaluate the potential aetiological role of prenatal events in childhood cancer. The obstetric records of 2692 mothers of children diagnosed with cancer and 4864 mothers of children without cancer were available for analysis. Overall, 1754 (65%) case mothers and 3220 (66%) control mothers had at least one prior pregnancy before the birth of the index child. Of these, 12 (0.68%) of the former and 9 (0.28%) of the latter had a prior molar pregnancy (odds ratio 2.5, 95% confidence interval 1.1 - 6.1). Both childhood cancer and molar pregnancy are rare neoplastic events, and the numbers are small. Nonetheless, whilst the associations were strongest for common precursor B-cell acute lymphoblastic leukaemia (OR 5.2, 95% CI 1.9 - 14.7) and sarcoma (OR 6.2, 95% CI 1.3 - 30.3), the spread across the remaining diagnostic groups suggests that the relationship, if confirmed, may be of a generalized, rather than specific, type. This is the first time that an association between childhood cancer and hydatidiform mole has been reported. The UKCCS's systematic use of clinical records permitted a more precise characterization of reproductive events than is possible in investigations that rely on individuals own accounts, and we are confident that our findings cannot be explained by recall bias or other methodological limitations. Accordingly, we suggest that there may be an aetiologic connection between molar pregnancy and childhood cancer, and speculate here on the various genetic/epigenetic mechanisms that could be involved.


Regulation of diaphragmatic oxygen uptake by endothelium-derived relaxing factor.
The role of endogenous nitric oxide (NO) in the regulation of phrenic blood flow (Qphr) and O2 consumption (VO2) of the in situ isolated left hemidiaphragms was assessed in two groups of anesthetized, mechanically ventilated dogs. Saline was infused into the phrenic artery for 20 min in one group, whereas N omega-nitro-L-arginine (L-NNA, 6 x 10(-4) M) was infused in the other (L-NNA) group. Qphr and diaphragmatic VO2 were measured at rest and during 2 min of continuous 3-Hz stimulation of the left phrenic nerve. The animals were progressively hemorrhaged, and the measurements were repeated at various arterial pressures (Pa). For the resting diaphragm, Qphr at a mean Pa of 145 mmHg was lower in the L-NNA group than in the saline group; however, diaphragmatic VO2 values were similar in both groups. Qphr decreased with the decline in Pa in both groups, but O2 extraction ratios obtained at mean Pa of 25-45 mmHg were similar in both groups (71 vs. 73%). For the contracting diaphragm, Qphr and diaphragmatic VO2 values at a given Pa were lower in the L-NNA group than in the saline group (except at mean Pa < 75 mmHg). O2 extraction ratios obtained at a given Pa were similar in both groups. We concluded that 1) EDRF inhibition limits diaphragmatic blood flow both at rest and during 3-Hz stimulation; 2) diaphragmatic O2 extraction is unaffected by EDRF inhibition; and 3) the effect of EDRF release on diaphragmatic VO2 is dependent on the level of metabolic demands.


Lowered anti-beta1 adrenergic receptor antibody concentrations may have prognostic significance in acute coronary syndrome.
Although several risk factors exist for acute coronary syndrome (ACS) no biomarkers for survival or risk of re-infarction have been validated. Previously, reduced serum concentrations of anti-ß1AR Ab have been implicated in poorer ACS outcomes. This study further evaluates the prognostic implications of anti-ß1AR-Ab levels at the time of ACS onset. Serum anti-ß1AR Ab concentrations were measured in randomly selected patients from within the PLATO cohort. Stratification was performed according to ACS event: ST-elevation myocardial infarct (STEMI) vs. non-ST elevation myocardial infarct (NSTEMI). Antibody concentrations at ACS presentation were compared to 12-month all-cause and cardiovascular mortality, as well as 12-month re-infarction. Sub-analysis, stratifying for age and the correlation between antibody concentration and conventional cardiac risk-factors was subsequently performed. Serum anti-ß1AR Ab concentrations were measured in 400/799 (50%) STEMI patients and 399 NSTEMI patients. Increasing anti-ß1AR Ab concentrations were associated with STEMI (p = 0.001). Across all ACS patients, no associations between anti-ß1AR Ab concentration and either all-cause cardiovascular death or myocardial re-infarction (p = 0.14) were evident. However among STEMI patients ≤60 years with anti-ß1AR Ab concentration <median higher rates of re-infarction were observed, compared to those with anti-ß1AR Ab concentrations > median (14/198 (7.1%) vs. 2/190 (1.1%)); p = 0.01). Similarly, the same sub-group demonstrated greater risk of cardiovascular death in year 1, including re-infarction and stroke (22/198 (11.1%) vs. 10/190 (5.3%); p = 0.017). ACS Patients ≤60 years, exhibiting lower concentrations of ß1AR Ab carry a greater risk for early re-infarction and cardiovascular death. Large, prospective studies quantitatively assessing the prognostic relevance of Anti-ß1AR Ab levels should be considered.


Avoidable blindness.
The vast majority of the world's 42 million blind are needlessly impaired. Epidemiologic studies are providing important insights into the cause of cataracts and provision of surgical services; ecologic approaches to the control of trachomatous corneal scarring; treatment and prevention of onchocerciasis; and early diagnosis and treatment of xerophthalmia among others. Continued research and application of existing knowledge can have a dramatic impact on the sight and lives of millions of people.


A novel de novo frameshift deletion in EHMT1 in a patient with Kleefstra Syndrome results in decreased H3K9 dimethylation.
Kleefstra Syndrome (KS) (MIM# 610253) is an autosomal dominant disorder caused by haploinsufficiency of euchromatic histone methyltransferase-1 (EHMT1, GLP). EHMT1 (MIM# 607001) encodes a histone methyltransferase that heterodimerizes with EHMT2 (also known as G9a, MIM# 604599), which together are responsible for mono- and dimethylation of H3 lysine 9 (H3K9me1 and -me2), resulting in transcriptional repression of target genes. This report describes an 18-year-old woman with intellectual disability, severely limited speech, hypotonia, microcephaly, and facial dysmorphisms, who was found to have a novel de novo single-base frameshift deletion in EHMT1. Functional studies using patient fibroblasts showed decreased H3K9me2 compared to wild-type control cells, thus providing a rapid confirmatory test that complements molecular studies. Whole exome sequencing revealed a novel frameshift deletion in EHMT1 after a lengthy diagnostic odyssey in this patient. Functional testing using this patient's fibroblasts provides proof-of-concept for the analysis of variants of uncertain significance that are predicted to impact EHMT1 enzymatic activity.


Localization and activation of the ARF6 GTPase during cleavage furrow ingression and cytokinesis.
The ARF6 GTPase mediates cell shape changes in interphase cells through its effects on membrane cycling and actin remodeling. In this study, we focus our attention on the dynamics of cell division and present evidence supporting a novel role for ARF6 during cleavage furrow ingression and cytokinesis. We demonstrate that endogenous ARF6 redistributes during mitosis and concentrates near the cleavage furrow during telophase. Constitutively activated ARF6 localizes to the plasma membrane at the site of cleavage furrow ingression and midbody formation, and dominant negative ARF6 remains cytoplasmic. By using a novel pull-down assay for ARF6-GTP, we find an abrupt, but transient, increase in ARF6-GTP levels as cells progress through cytokinesis. Whereas high levels of expression of a GTPase-defective ARF6 mutant induce aberrant phenotypes in cells at cytokinesis, cells expressing low levels of ARF6 mutants do not display a significant mitotic delay or cytokinesis defect, presumably due to compensatory or redundant mechanisms that allow cytokinesis to proceed when the ARF6 GTPase cycle is disrupted. Finally, actin accumulation and phospholipid metabolism at the cleavage furrow are unchanged in cells expressing ARF6 mutants, suggesting that ARF6 may be involved in membrane remodeling during cytokinesis via effector pathways that are distinct from those operative in interphase cells.


Review article: type 2 diabetes and chronic liver disease in the Verona diabetes study.
Non-alcoholic liver steatosis is associated with metabolic syndrome and type 2 diabetes. The prevalence of this condition in type 2 diabetes is estimated to be between 28 and 55%. Non-alcoholic liver steatosis is not a benign disease because of its potential progression to liver fibrosis, cirrhosis and cancer. The Verona diabetes study, a population-based observational study, on 7148 type 2 diabetic patients after 5 years of follow-up has reported an increased risk of death from gastrointestinal diseases, particularly from chronic liver cirrhosis. Moreover, in the same population after 10 years of follow-up a higher risk of mortality from liver cancer was observed and this risk increased significantly in obese patients (body mass index >30 kg/m2). Of note is that obese diabetic patients suffer an even higher prevalence of non-alcoholic liver steatosis. In conclusion, the Verona diabetes study showed an increased risk of mortality from liver cirrhosis and liver cancer in type 2 diabetic patients. Diverse pathophysiological mechanisms can be responsible, i.e. higher alcohol consumption, hepatitis and others but, considering the high prevalence of non-alcoholic liver steatosis in these patients, it is plausible to hypothesize that non-alcoholic liver steatosis may play a significant role in predisposing the liver of diabetics to chronic diseases.


Genetic and physical localization of an anthracnose resistance gene in Medicago truncatula.
Anthracnose of alfalfa, caused by the fungal pathogen Colletotrichum trifolii, is one of the most destructive diseases of alfalfa worldwide. An improved understanding of the genetic and molecular mechanisms underlying host resistance will facilitate the development of resistant alfalfa cultivars, thus providing the most efficient and environmentally sound strategy to control alfalfa diseases. Unfortunately, cultivated alfalfa has an intractable genetic system because of its tetrasomic inheritance and out-crossing nature. Nevertheless, the model legume Medicago truncatula, a close relative of alfalfa, has the potential to serve as a surrogate to map and clone the counterparts of agronomically important genes in alfalfa -- particularly, disease resistance genes against economically important pathogens. Here we describe the high-resolution genetic and physical mapping of RCT1, a host resistance gene against C. trifolii race 1 in M. truncatula. We have delimited the RCT1 locus within a physical interval spanning approximately 200 kb located on the top of M. truncatula linkage group 4. RCT1 is part of a complex locus containing numerous genes homologous to previously characterized TIR-NBS-LRR type resistance genes. The result presented in this paper will facilitate the positional cloning of RCT1 in Medicago.


Primary follicular lymphoma of the duodenum with erosions as atypical macroscopic features.
A 52-year-old Japanese woman who was eventually diagnosed with primary follicular lymphoma of the duodenum showed atypical endoscopic features, namely, erosions with peripheral whitish edematous mucosa. Initial biopsy specimens taken from the erosions revealed insufficient numbers of lymphoma cells for histological diagnosis. Subsequent biopsy specimens from the peripheral mucosa containing the whitish enlarged villi showed infiltration of the lymphoma cells forming lymphoid follicles, which led us to the appropriate diagnosis. This case indicates that endoscopists should take biopsy samples from the peripheral mucosa with whitish enlarged villi rather than erosions in the rare instances that erosions appear as the main macroscopic feature of intestinal follicular lymphoma.


Alteration in TRPV1 and Muscarinic (M3) receptor expression and function in idiopathic overactive bladder urothelial cells.
To examine function of both cholinergic (muscarinic) and TRPV1 receptors in human bladder urothelial (HBUC) from non-neurogenic overactive bladder (OAB) patients as compared to control subjects. Primary HBUC cultures were derived from cystoscopic biopsies from OAB and control subjects. Muscarinic and TRPV1 function was assessed by acetylcholine (5 μm) or capsaicin (0.5 μm) evoked ATP release, measured by luciferase assay. Overall, expression of TRPV1 and muscarinic M3 receptors in bladder urothelial cells was accomplished using western immunoblotting. Our findings revealed that the response to acetylcholine in OAB HBUC cultures (which was blocked by the nonselective muscarinic antagonist, atropine methyl nitrate or AMN) was not significantly different than from controls. The acetylcholine M3 receptor was slightly decreased as compared to control. In contrast, OAB HBUC cultures exhibited a capsaicin hypersensitivity and augmented release of ATP (3.2 fold higher), which was blocked by the antagonist capsazepine. The increase in capsaicin sensitivity correlated with increased urothelial TRPV1 expression. Though characterized in a small number of subjects, augmented release of urothelial-derived transmitters such as ATP could 'amplify' signalling between and within urothelial cells and nearby afferent nerves.


Applying genomics in heart transplantation.
While advances in patient care and immunosuppressive pharmacotherapies have increased the lifespan of heart allograft recipients, there are still significant comorbidities post-transplantation and 5-year survival rates are still significant, at approximately 70%. The last decade has seen massive strides in genomics and other omics fields, including transcriptomics, with many of these advances now starting to impact heart transplant clinical care. This review summarizes a number of the key advances in genomics which are relevant for heart transplant outcomes, and we highlight the translational potential that such knowledge may bring to patient care within the next decade.


Distribution of Carbapenemases and Efflux Pump in Carbapenem-resistance Acinetobacter baumannii.
Acinetobacter baumannii has emerged as an important pathogen related to serious infections and nosocomial outbreaks around the world. In this study, 100 isolates of carbapenem-resistance in Acinetobacter baumannii (CRAB) were collected from clinical specimens. Agar dilution was conducted to determine the minimum inhibitory concentrations (MICs) for 15 kinds of antibiotics. Genes of carbapenemases and efflux pumps were amplified by PCR. The expression difference of pump genes was analyzed by real-time PCR between CRAB and carbapenems-sensitive Acinetobacter baumannii (CSAB). We found that most antibiotics, including aminoglycosides, fluoroquinolones, and cephalosporins showed high MIC values in CRAB. All isolates were sensitive to polymyxin B. Among the CRAB specimens, 54, 32 and 16 isolates were positive for SHV-12, PER-1 and TEM-1, respectively. 86 isolates were positive for OXA-23. 55 and 33 isolates carried adeB and adeJ genes, respectively. The expression level of adeB in CRAB was ten times higher than that in CSAB. We speculate that SHV-12, PER-1, TEM-1, OXA-23 and the AdeABC efflux pump may participate in high-level carbapenems resistance in Acinetobacter baumannii Moreover, adeE may be related to low-level resistance of carbapenems and quinolones in Acinetobacter baumannii.


[A case of intussusception caused by descending colon cancer in a very elderly patient, lacking in abdominal symptoms].
A 90-year-old woman was admitted to our hospital because of dizziness, without any remarkable abdominal symptoms. Severe anemia was found and thus gastrointestinal investigations were performed. CT and ultrasonography showed the specific multiplex layer configuration at the anal side of the descending colon. Colonoscopy showed a round tumor occupying the lumen and a consecutive gastrografin enema showed a crab-claw-like area without filling in the descending colon. Intussusception of the colon was diagnosed and partial resection of the descending colon was performed, but without relief of intussusception. The tumor was histologically diagnosed as mucinous adenocarcinoma. By various factors, incomplete fixation between the retroperitoneum and the descending colon might result in the present condition.


Oral hygiene and gingival health in patients with fixed prosthodontic appliances--a 12-month follow-up.
The aim of this study was to assess and observe the oral hygiene and gingival condition in patients before and after fixed prosthodontic therapy through a 12-month period in combination with oral hygiene instructions. It was also analysed how factors, such as type of fixed prosthodontic appliance, constructive material, the position of a fixed partial denture (FPD) in the mouth, age and gender influenced oral hygiene. The dental arches were divided into three segments each and teeth and gingiva were examined using the Plaque and Gingiva Index by Silness and Löe, and for the mineralized deposits assessment the Calculus Index by Green and Vermillion was employed. The preliminary examination was conducted before the prosthodontic therapy, and the reexaminations were carried out 14 days, 1, 6 and 12 months after crown and/or FPD placement. A total of 93 subjects from the original study group of 146 patients attended all clinical examinations, while the rest was excluded. The sample consisted of 60 women, 33 men at age between 21 and 95 (average 51.8). A total of 39 patients had single crowns (C), 50 FPDs and 5 C+FPD. The frequency of plaque found during the preliminary visit was higher than that found in the other periods (p < 0.001). Patients with C showed better oral hygiene levels than patients with FPDs or C+FPDs (p = 0.001). Our results revealed no significant difference in oral hygiene status among patients with FPDs made of different materials (p = 0.083). The worst hygiene levels were found in patients with fixed prosthodontic appliances in both jaws (p = 0.012). Younger patients showed better hygiene levels than the older ones (p = 0.002). Our research showed that appropriate educational and motivational measures can lead to improved oral hygiene, even after FPD placement. Presumably, the oral health in a group of adult patients can be kept acceptable by providing a prophylactic oral hygiene program.


The strategy of diminishing age gap effect on different donor-recipient combinations in living donor kidney transplantation.
The disparity between kidney donation and the number of uremic patients on the waiting list has increased the demand for older live-donor kidneys (OLK). However, the donor-recipient age gap may have an impact on the recipient's outcome. Patients who underwent living donor kidney transplantation at our institute between 2005 and 2019 were enrolled and categorized into four donor-recipient groups according to age (≥50 years and <50 years). The Estimated Post-Transplant Survival (EPTS) score was used to quantify the recipient's condition. Adjusted models analyzed recipient outcomes and related risks among the four groups. Of the 154 pairs of live donors and recipients, OLK did not influence overall or death-censored graft survival. The four donor-recipient combinations had similar recipient outcomes, except it slightly worsened in the "old donor to young recipient" group. The EPTS score (adjusted HR, 1.02; 95% CI, 1.01-1.04; p = 0.014) and rejection (adjusted HR, 4.26; 95% CI, 1.36-13.37; p = 0.013) were significant risk factors for overall and death-censored graft survival, respectively. Recipients with pretransplant diabetes or prior solid organ transplantation could have amplified risk effects. The main causes of graft loss were death in older recipients and chronic rejection in younger recipients. OLK is safe for young recipients. Nevertheless, adequate immunosuppression should be maintained to prevent rejection and subsequent graft loss, especially for those receiving second kidney transplantation. In contrast, older recipients should avoid overt immunosuppression and control their comorbidities, such as diabetes-related complications to improve their long-term outcomes.


High-affinity σ1 protein agonist reduces clinical and pathological signs of experimental autoimmune encephalomyelitis.
Selective agonists of the sigma-1 receptor (σ1 protein) are generally reported to protect against neuronal damage and modulate oligodendrocyte differentiation. Human and rodent lymphocytes possess saturable, high-affinity binding sites for compounds binding to the σ1 protein and potential immunomodulatory properties have been described for σ1 protein ligands. Experimental autoimmune encephalomyelitis (EAE) is recognized as a valuable model of the inflammatory aspects of multiple sclerosis (MS). Here, we have assessed the role of a σ1 protein agonist, containing the tetrahydroisoquinoline-hydantoin structure, in EAE. EAE was induced in SJL/J female mice by active immunization with myelin proteolipid protein (PLP)139-151 peptide. The σ1 protein agonist was injected i.p. at the time of immunization (day 0). Disease severity was assessed clinically and by histopathological evaluation of the CNS. Phenotyping of B-cell subsets and regulatory T-cells were performed by flow cytometry in spleen and cervical lymph nodes. Prophylactic treatment of EAE mice with the σ1 protein agonist prevented mononuclear cell accumulation and demyelination in brain and spinal cord and increased T2 B-cells and regulatory T-cells, resulting in an overall reduction in the clinical progression of EAE. This σ1 protein agonist, containing the tetrahydroisoquinoline-hydantoin structure, decreased the magnitude of inflammation in EAE. This effect was associated with increased proportions of B-cell subsets and regulatory T-cells with potential immunoregulatory functions. Targeting of the σ1 protein might thus provide new therapeutic opportunities in MS.


[A preliminary analysis of changes in composition of intestinal microbiota during infancy using polymerase chain reaction-denaturing gradient gel electrophoresis].
To investigate the composition of bacteria in the stools of infants and the colonization of intestinal microbiota during infancy. Fresh stools were collected from 15 healthy infants at 0, 2, 4, 7, 10, 14, and 28 days and 3, 6, and 12 months after birth. Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) was used to analyze the composition of intestinal microbiota, perform sequencing of dominant bacteria, and to analyze the changes in the composition of intestinal microbiota during infancy. DGGE fingerprint showed that the composition of intestinal microbiota during infancy changed significantly over time after birth. The cloning and sequencing results indicated that Proteobacteria colonized the earliest, mainly the obligate aerobes Enterobacter and Pseudomonas, followed by the obligate anaerobes (Clostridium hathewayi and Veillonella parvula) and the facultative anaerobe Clostridium ramosum in Firmicutes, and Verrucomicrobia. Actinobacteria colonized the latest, mainly Bifidobacterium, and gradually became dominant bacteria. During infancy, obligate aerobes colonize the intestinal tract the earliest, followed by obligate anaerobes and facultative anaerobes. Proteobacteria colonizes the earliest, followed by Firmicutes and Verrucomicrobia, and Actinobacteria, mainly Bifidobacterium, colonizes the latest.


Effect of variations in small intestinal glucose delivery on plasma glucose, insulin, and incretin hormones in healthy subjects and type 2 diabetes.
The determinants of postprandial blood glycemia are controversial. We assessed the effects of variations in the initial rate of small intestinal glucose delivery on blood glucose, plasma insulin, and incretin responses in both health and type 2 diabetes. Eight controls and eight patients with type 2 diabetes managed by diet alone underwent paired studies. On both days subjects received an intraduodenal glucose infusion (t = 0-120 min); on one day the infusion rate was variable, being more rapid initially (3 kcal/min) between t = 0 and 15 min and slower (0.71 kcal/min) subsequently (t = 15-120 min), whereas on the other day, the infusion rate was constant (1 kcal/min) from t = 0 to 120 min (i.e. on both days 120 kcal of glucose were administered). Between t = 0-180 min blood glucose, plasma insulin and plasma glucose-dependent insulin-releasing polypeptide were greater with the variable, compared with the constant, infusion. Between t = 0 and 30 min the magnitude of the rise in plasma glucagon-like peptide-1 was greater with the variable, compared with the constant infusion (P < 0.01, both groups). We conclude that modest variations in the initial rate of duodenal glucose entry may have profound effects on subsequent glycemic, insulin, and incretin responses.


Mast Cells Promote Seasonal White Adipose Beiging in Humans.
Human subcutaneous (SC) white adipose tissue (WAT) increases the expression of beige adipocyte genes in the winter. Studies in rodents suggest that a number of immune mediators are important in the beiging response. We studied the seasonal beiging response in SC WAT from lean humans. We measured the gene expression of various immune cell markers and performed multivariate analysis of the gene expression data to identify genes that predict UCP1. Interleukin (IL)-4 and, unexpectedly, the mast cell marker CPA3 predicted UCP1 gene expression. Therefore, we investigated the effects of mast cells on UCP1 induction by adipocytes. TIB64 mast cells responded to cold by releasing histamine and IL-4, and this medium stimulated UCP1 expression and lipolysis by 3T3-L1 adipocytes. Pharmacological block of mast cell degranulation potently inhibited histamine release by mast cells and inhibited adipocyte UCP1 mRNA induction by conditioned medium (CM). Consistently, the histamine receptor antagonist chlorpheniramine potently inhibited adipocyte UCP1 mRNA induction by mast cell CM. Together, these data show that mast cells sense colder temperatures, release factors that promote UCP1 expression, and are an important immune cell type in the beiging response of WAT.


Perinatal folate levels do not influence tumor latency or multiplicity in a model of NF1 associated plexiform-like neurofibromas.
In epidemiological and experimental research, high folic acid intake has been demonstrated to accelerate tumor development among populations with genetic and/or molecular susceptibility to cancer. Neurofibromatosis type 1 (NF1) is a common autosomal dominant disorder predisposing affected individuals to tumorigenesis, including benign plexiform neurofibromas; however, understanding of factors associated with tumor risk in NF1 patients is limited. Therefore, we investigated whether pregestational folic acid intake modified plexiform-like peripheral nerve sheath tumor risk in a transgenic NF1 murine model. We observed no significant differences in overall survival according to folate group. Relative to controls (180 days), median survival did not statistically differ in deficient (174 days, P = 0.56) or supplemented (177 days, P = 0.13) folate groups. Dietary folate intake was positively associated with RBC folate levels at weaning, (P = 0.023, 0.0096, and 0.0006 for deficient vs. control, control vs. supplemented, and deficient vs. supplemented groups, respectively). Dorsal root ganglia (DRG), brachial plexi, and sciatic nerves were assessed according to folate group. Mice in the folate deficient group had significantly more enlarged DRG relative to controls (P = 0.044), but no other groups statistically differed. No significant differences for brachial plexi or sciatic nerve enlargement were observed according to folate status.


New applications of intracardiac echocardiography: assessment of coronary blood flow by colour and pulsed Doppler imaging in dogs.
To explore the application of a new 10 French intracardiac echocardiography (ICE) catheter with phased array and Doppler capable transducer for the assessment of epicardial and intramyocardial coronary blood flow. The coronary arteries were detected by cross sectional imaging in seven closed chest dogs, and coronary blood flow visualised by colour Doppler. Blood flow velocities were recorded by pulsed Doppler at baseline for reproducibility of repeated measurements, and during hyperaemia for coronary flow reserve measurements. Comparisons were made with Doppler guide wire data obtained simultaneously. Intramyocardial coronary artery blood flow was assessed by colour flow mapping, and the blood flow velocities recorded using pulsed Doppler at baseline and during hyperaemia. Seven left main, six left anterior descending, seven left circumflex, and five right coronary arteries were visualised in the seven animals by cross sectional or colour Doppler imaging. Repeated measurements of coronary flow velocity showed a good correlation (mean diastolic velocity, r = 0.93, n = 22, p < 0.0001; peak diastolic velocity, r = 0.96, n = 22, p < 0.0001, respectively). Intraobserver/interobserver variability was satisfactorily low. Coronary flow reserve from ICE correlated highly with the value obtained from the Doppler guide wire (r = 0.90, n = 26, p < 0.0001). Intramyocardial coronary blood flow was identified in all seven dogs, and flow velocities were recorded at baseline and during hyperaemia in four animals. This new ICE catheter provides high quality diagnostic resolution. It is useful for coronary blood flow assessment.


The Making of the Andrea Wave and other Rogues.
Unexpectedly large ocean waves or 'rogues' are sometimes claimed to be the cause of damage to ships at sea and to offshore structures. While wind-driven wave models are capable of predicting the average characteristics of waves, the maximum height of rogues that may occur is yet unknown. Rogues form in the open ocean through the addition of elemental wave trains or groups and, infrequently, with many elements coming together in phase, producing rogues. Here we perform directional analyses on one of the steepest rogues ever recorded: the Andrea wave. We find that the Andrea wave was close to the breaking-limited height. Analysis of the 72 twenty minute records on the day of the Andrea wave yields encounter return periods of about 21 days for maximally steep waves, while less steep rogues occur about twice daily. An explicit formula is given for the encounter probability, based on the target area. This work answers the critical questions regarding rogues in the design and operation of ships and offshore structures: how high can rogues be and how frequently they occur.


Combined association of clinical and lifestyle factors with non-restorative sleep: The Nagahama Study.
Non-restorative sleep (NRS) was suggested to be associated with cardiovascular outcomes. However, causative factors for NRS have not been fully elucidated. This study aimed to clarify factors and their relationships with NRS to better understand the clinical and epidemiological implications of NRS and to develop a score that can objectively evaluate NRS status. Study subjects consisted of 9,788 community residents (age 53.6 ± 13.4 y). Subjective NRS as well as possible clinical and lifestyle factors for NRS were investigated by questionnaires. Other clinical parameters were obtained from personal records of information obtained at the baseline examination. A total of 3,261 participants complained of NRS. Factors independently associated with subjective NRS were younger age (odds ratio = 1.43), use of a hypnotic drug (2.04), irregular sleep schedule (2.02), short sleep duration (<5 h, 11.7; 5-6 h, 4.81; 6-7 h, 2.40), frequent sleepiness (2.33), routine stress (4.63), no habitual exercise (1.61), nocturia symptoms (1.43), symptoms of gastroesophageal reflux disease (1.44), and depression (1.46) (all P <0.001). The NRS score comprised of these 10 factors was linearly associated with the frequency of subjective NRS (Ptrend <0.001). Frequency of individuals with a high NRS score was greater in women (52.3%) than in men (42.1%, P<0.001), while no clear association was observed with common risk factors for cardiovascular diseases. NRS was a phenomenon representing various clinical and lifestyle features. Careful attention should be paid to individuals with a high NRS score who might be at risk for mental fatigue and have unfavorable lifestyle factors.


Cyclin-dependent kinase 1 (CDK1)-mediated mitotic phosphorylation of the transcriptional co-repressor Vgll4 inhibits its tumor-suppressing activity.
The Hippo pathway is an evolutionarily conserved signaling pathway that plays important roles in stem cell biology, tissue homeostasis, and cancer development. Vestigial-like 4 (Vgll4) functions as a transcriptional co-repressor in the Hippo-Yes-associated protein (YAP) pathway. Vgll4 inhibits cell proliferation and tumor growth by competing with YAP for binding to TEA-domain proteins (TEADs). However, the mechanisms by which Vgll4 itself is regulated are unclear. Here we identified a mechanism that regulates Vgll4's tumor-suppressing function. We found that Vgll4 is phosphorylated in vitro and in vivo by cyclin-dependent kinase 1 (CDK1) during antimitotic drug-induced mitotic arrest and also in normal mitosis. We further identified Ser-58, Ser-155, Thr-159, and Ser-280 as the main mitotic phosphorylation sites in Vgll4. We also noted that the nonphosphorylatable mutant Vgll4-4A (S58A/S155A/T159A/S280A) suppressed tumorigenesis in pancreatic cancer cells in vitro and in vivo to a greater extent than did wild-type Vgll4, suggesting that mitotic phosphorylation inhibits Vgll4's tumor-suppressive activity. Consistent with these observations, the Vgll4-4A mutant possessed higher-binding affinity to TEAD1 than wild-type Vgll4. Interestingly, Vgll4 and Vgll4-4A markedly suppressed YAP and β-catenin signaling activity. Together, these findings reveal a previously unrecognized mechanism for Vgll4 regulation in mitosis and its role in tumorigenesis.


Real-world effectiveness of sucroferric oxyhydroxide in patients on chronic hemodialysis: A retrospective analysis of pharmacy data .
Hyperphosphatemia has been associated with an increased risk of mortality in patients with end-stage renal disease. We sought to assess the real-world effectiveness of sucroferric oxyhydroxide (SO), an iron-based phosphate binder (PB), in control of serum phosphorus levels, and to determine the associated pill burden in hemodialysis patients. Adult, in-center hemodialysis patients first prescribed SO through a renal pharmacy service as part of routine clinical care between April 1, 2014 and March 31, 2015 were included in the analysis. The proportion of patients with phosphorus levels ≤ 5.5 mg/dL and the mean prescribed PB pills/day were compared between baseline (3 months prior to SO) and SO follow-up at 3 (SO 1 - 3) and 6 months (SO 4 - 6). Mineral bone disease markers, hemoglobin, iron indices, and erythropoiesis-stimulating agents and intravenous iron use were assessed. At baseline, all patients (n = 1,029) were prescribed PB, and 13.9% had mean serum phosphorus ≤ 5.5 mg/dL. Comparing baseline to SO 1 - 3, the mean prescribed PB pills/day declined from 9.6 to 3.8 pills/day (p < 0.001), and the proportion of patients with serum phosphorus ≤ 5.5 mg/dL increased from 13.9 to 26.1% (+88%). Comparing baseline to SO 4 - 6 (n = 424), the mean prescribed PB pills/day declined from 9.7 to 4.0 pills/day (p < 0.001), and the proportion of patients with serum phosphorus ≤ 5.5 mg/dL increased from 15.6 to 30.4% (+95%). Prescription of SO was associated with an increase in the proportion of patients achieving serum phosphorus levels ≤ 5.5 mg/dL along with fewer prescribed PB pills/day. .


The role of CD4 and CD8 T cells in the graft-versus-leukemia response in Rauscher murine leukemia.
Using a mouse model for MHC-matched unrelated donor transplantation, the relative influences of the CD4 and CD8 T cell subtypes on graft-versus-leukemia (GVL) were examined in a murine erythroleukemia induced in SJL/J mice by the injection of Rauscher virus. Following leukemia induction, the mice were given 9.5 Gy of total body irradiation (TBI) and injected with mixed marrow and spleen cells from normal MHC-matched--but minor histocompatibility mismatched--B10.S donors. Prior to their injection these donor cells were selectively depleted ex vivo for either CD4, CD8 or Thy-1 by exposure to the appropriate monoclonal antibody (MoAb) plus complement. Following transplant the recipients were observed for 20 weeks, along with parallel control groups, for survival, leukemia relapse, graft failure and graft-versus-host disease; 98% of the controls receiving no transplantation therapy died of leukemia. Among the controls that received TBI plus undepleted B10.S cells 30.9% died of leukemia relapse, but another 34.2% survived free of any clinical evidence of their leukemia. Donor cell depletion for Thy-1 increased the relapse to 68.8%, while survival fell to 10.4%. CD8 depletion resulted in a relapse of 55.6%, with a survival of 19.4%. By contrast, CD4 depletion had no effect on relapse, but did significantly increase the incidence of graft failure. At the end of the 20 weeks additional tests were run to determine whether those transplant survivors that had remained leukemia-free were also free of any residual Rauscher virus. Those tests showed that they were not.(ABSTRACT TRUNCATED AT 250 WORDS)


Enantioselective organocatalytic approach to the synthesis of alpha,alpha-disubstituted cyanosulfones.
Optically pure cyano tert-alkyl sulfones have been obtained by organocatalytic enantioselective Michael addition of alpha-substituted cyanosulfones to vinyl ketones using cinchona alkaloids as catalysts. The best results were obtained for p-trifluorophenylsulfones by using VIII as catalyst in toluene at -40 degrees C. Reactions proved to be applicable for a variety of alpha,beta-unsaturated ketones, affording alpha,alpha-disubstituted cyanosulfones in excellent yields with er's up to 90:10.


Germline mutations in BRIP1 and PALB2 in Jewish high cancer risk families.
Germline mutations in BRCA1 and BRCA2 account for ~30 % of inherited breast cancer. BRIP1 and PALB2 are likely genes for breast cancer susceptibility, based on their roles in maintaining cellular integrity. Indeed, few pathogenic germline mutations in both genes are reported in ethnically diverse breast cancer families. There is a paucity of data on the putative contribution of both genes to inherited breast cancer in Jewish high risk families. High risk Jewish women, none of whom was a carrier of the predominant Jewish mutations in BRCA1/BRCA2, were screened for BRIP1 germline mutations by combined denaturing gradient gel electrophoresis, high resolution melting and sequencing. Direct sequencing of exons and flanking intronic sequences was used for PALB2 mutational analysis. Overall, 149 women, all of high risk, cancer prone families of Ashkenazi origin, were genotyped for BRIP1 mutations: 127 with breast cancer, 22 with ovarian cancer. No truncating mutations were noted and one novel (p.Ala745Thr) and two previously described missense mutations were detected. For PALB2, 93 women were genotyped (87 with breast cancer) of Ashkenazi (n = 32) and non Ashkenazi Jewish origin. Fifteen sequence variants were detected, of these, none was truncating, four were not previously reported, and two (p.Asp871Gly and p.Leu1119Pro) were seemingly pathogenic based on the PolyPhen2 protein prediction algorithm. These missense mutations were not detected in any of 113 healthy Ashkenazi and 109 Moroccan, cancer free controls. In conclusion, germline mutations in BRIP1 and PALB2 contribute marginally to breast cancer susceptibility in ethnically diverse, Jewish high risk families.


Comparative secretome analysis of human bone marrow-derived mesenchymal stem cells during osteogenesis.
Osteogenesis is a tightly regulated process that involves coordinated extracellular signals from autocrine and paracrine loops. Secretory proteins during osteogenesis can inhibit cell proliferation and activate cell differentiation toward mature osteoblasts, which are characterized by mineralization. In this study, we attempted to identify these secretory proteins during osteogenesis using LC-MS/MS analysis. We compared the secretome between undifferentiated human bone marrow-derived mesenchymal stem cells (hBMSCs) and differentiated osteoblasts. Among 315 proteins that were identified, 177 proteins were present at increased levels in osteoblasts, whereas 88 proteins were present at decreased levels. Among the identified proteins, several were validated by quantitative RT-PCR and immunoblot analysis. Of particular interest, calcium homeostasis-related proteins were upregulated, whereas stem cell proliferation-related proteins and other lineage-related proteins were downregulated during osteogenesis. These findings provide information about the dynamic changes in the expression and secretion of proteins during osteogenesis and suggest the putative role of secretory proteins in osteogenesis.


From nanofabrication to self-fabrication--tailored chemistry for control of single molecule electronic devices.
Single molecule electronics is a field of research focused on the use of single molecules as electronics components. During the past 15 years the field has concentrated on development of test beds for measurements on single molecules. Bottom-up approaches to single molecule devices are emerging as alternatives to the dominant top-down nanofabrication techniques. One example is solution-based self-assembly of a molecule enclosed by two gold nanorod electrodes. This article will discuss recent attempts to control the self-assembly process by the use of supramolecular chemistry and how to tailor the electronic properties of a single molecule by chemical design.


Cellular proteomic analysis of porcine circovirus type 2 and classical swine fever virus coinfection in porcine kidney-15 cells using isobaric tags for relative and absolute quantitation-coupled LC-MS/MS.
Viral coinfection or superinfection in host has caused public health concern and huge economic losses of farming industry. The influence of viral coinfection on cellular protein abundance is essential for viral pathogenesis. Based on a coinfection model for porcine circovirus type 2 (PCV2) and classical swine fever virus (CSFV) developed previously by our laboratory, isobaric tags for relative and absolute quantitation (iTRAQ)-coupled LC-MS/MS proteomic profiling was performed to explore the host cell responses to PCV2-CSFV coinfection. Totally, 3932 proteins were identified in three independent mass spectrometry analyses. Compared with uninfected cells, 304 proteins increased (fold change >1.2) and 198 decreased (fold change <0.833) their abundance in PCV2-infected cells (p < 0.05), 60 and 61 were more and less abundant in CSFV-infected cells, and 196 and 158 were more and less abundant, respectively in cells coinfected with PCV2 and CSFV. Representative differentially abundant proteins were validated by quantitative real-time PCR, Western blotting and confocal laser scanning microscopy. Bioinformatic analyses confirmed the dominant role of PCV2, and indicated that mitochondrial dysfunction, nuclear factor erythroid 2-related factor 2 (Nrf2)-mediated oxidative stress response and apoptosis signaling pathways might be the specifical targets during PCV2-CSFV coinfection.


A systematic review of the outcomes of cardiovascular exercise programs for people with Down syndrome.
To determine if cardiovascular exercise programs are beneficial and safe for people with Down syndrome. Electronic databases were searched from the earliest time available through to October 2004 using the following key words: Down syndrome or trisomy 21 in combination with physical fitness, exercise, physical activity, exercise therapy, exercise training, physical training, and aerobic. Additional articles were identified by manual searching and citation tracking. Two reviewers independently assessed the articles identified in the initial search for the following inclusion criteria: (1) participants with Down syndrome, (2) an exercise program that conformed with the American College of Sports Medicine guidelines for increasing cardiovascular fitness, (3) assessed changes in body structure or function, activity limitation, or societal participation, and (4) used a prospective clinical controlled research design with or without random allocation to groups. Trials of low methodologic quality were excluded (PEDro score, <4). Of the 156 articles initially identified, only 4 met the inclusion criteria and underwent detailed review. Data relating to changes in body structure and function, activity limitation, participation restriction, and contextual factors from the included studies were independently extracted by the reviewers on a standardized form. Study quality was assessed using the PEDro scale. Meta-analyses found that cardiovascular exercise programs were effective in increasing peak oxygen consumption (d=.75; 95% confidence interval [CI], 0.34-1.15), peak minute ventilation (d=.71; 95% CI, 0.15-1.28), the maximum workload achieved (d=.96; 95% CI, 0.45-1.45), and the time to exhaustion (d=.72; 95% CI, 0.29-1.15) in people with Down syndrome. No changes were found for body weight (d=.09; 95% CI, -.39 to .57). No adverse effects were reported in any of the studies. The result of our review support the use of programs designed to improve cardiovascular fitness among people with Down syndrome. However, because only 4 studies were included, the findings need to be interpreted cautiously. High-quality randomized controlled trials should be completed in the future to determine the effect of these programs on activity and social participation.


Evaluation of vestibular evoked myogenic potentials.
In previous studies, electromyographic potentials, recorded in response to auditory clicks, have been attributed to stimulation of the otolith (saccule) and have been termed vestibular evoked myogenic potentials (VEMPs). In this study, we assessed the VEMPs in subjects with normal auditory brainstem evoked responses, with no history of vestibular symptoms or neck and other skeletal muscle abnormalities. To this effect, 32 subjects (64 ears), after ethics committee approval, were exposed to 75, 150, and 300 clicks at 100 dB, and the responses were averaged. Electromyographic activity was recorded by applying surface electrodes over the sternocleiodomastoid muscle under the following three conditions: no muscle contraction/no clicks, muscle contraction/no clicks, and muscle contraction/clicks. Our findings suggest that electromyographic responses have to be obtained, during muscle contraction, first without and then with clicks. Our data also suggest that comparison of these two recordings is necessary for meaningful results.


Use of animals in biomedical research. Historical role of the American Medical Association and the American physician.
From the introduction of the "Gallinger-DC" bill in 1896 to the passage of the Laboratory Animal Welfare Act in 1966, organized medicine and the American physician have been active in promoting the humane and appropriate use of research animals and explaining to the public and legislators the importance of research using animals to medical progress. The role of organized medicine and science in events leading to passage of federal legislation is discussed. Past efforts of the American Medical Association and the American physician have been critical in numerous successful efforts at the local, state, and national level to prevent the passage of laws which restricted animal use for health research and impeded medical progress. This article demonstrates that current initiatives by physicians to preserve biomedical research are a reaffirmation of their traditional role.


Cancer in Jewish migrants to Israel.
Data on the incidence of 15 cancers in the Jewish population of Israel from the period 1961-1981 have been studied with emphasis on the risk of disease in relation to birthplace, calendar time, and duration of residence in Israel. The results show wide variations in incidence by birthplace, from less than 2-fold for pancreatic cancer to a 40-fold difference for oesophageal cancer in females; the reasons for these are often little understood. The potential importance of environmental agents in aetiology for different migrant groups is discussed in relation to the size and rapidity of changes in risk related to the duration of residence in Israel.


Tissue and plasma enzyme activities and chemical analytes in Golden Trevally from a public aquarium.
Veterinary care of aquatic species, particularly fish, is limited by a lack of knowledge regarding their unique physiology. Tissue enzyme activities measured in plasma are used for assessing function and potential damage to specific organs and tracking disease progression in live animals. The objective of this study was to identify tissue(s) of origin and plasma concentrations for specific enzymes in healthy Golden Trevally Gnathanodon speciosus. We hypothesized that enzymes would exhibit tissue-specific tropisms, with higher activities in one or more tissues compared to others. Six fish were randomly selected from a public aquarium population to obtain antemortem blood samples. The fish were then euthanized, and tissue samples were collected via gross necropsy. Six enzyme activities and two chemical analytes were examined across samples of plasma and 10 tissues from each fish. Enzyme activities exhibited significant organ specificities. Aspartate aminotransferase, lactate dehydrogenase, and creatine kinase levels were highest in skeletal muscle, with variably high levels in gonads. Alkaline phosphatase levels were highest in the kidney, spleen, and liver. Alanine aminotransferase levels had high specificity for the liver. Gamma-glutamyl transferase was only detectable in the kidney and plasma. This work establishes baseline tissue enzyme origins for Golden Trevally, which will aid clinicians in diagnostic interpretation of blood chemistries and improve veterinary care for this understudied fish species.


Effect of Recurrent Mitral Regurgitation After Mitral Valve Repair in Patients With Degenerative Mitral Regurgitation.
This study investigated the consequences of recurrent mitral regurgitation (MR) after mitral valve (MV) repair in patients with degenerative MR and risk factors for recurrence.Methods and Results:From January 1990 to December 2015, 792 patients underwent MV repair due to degenerative MR. Recurrent MR was defined as moderate-to-severe MR on follow-up echocardiography. Mean follow-up duration was 8.71±5.58 years. During the follow-up period, MR recurred in 133 (16.8%) patients, and the MR recurrence-free rate at 20 years was 77.5±2.0%. In the recurrence group, the degree of MR decreased in 8 (6.0%) patients and was aggravated in 46 (34.6%) patients. Recurrent MR was associated with increased mortality and adverse left ventricular (LV) remodeling. Independent risk factors for MR recurrence were MV repair performed before 2000, preoperative atrial fibrillation, high LV end-diastolic dimension (LVEDD), prolapse of the isolated anterior leaflet or multiple segments, and absence of ring annuloplasty. Predictors of MR progression were high LVEDD and repair without artificial chordae implantation. Recurrent MR after MV repair in patients with degenerative MR showed a tendency to progress and was associated with increased mortality and adverse LV remodeling. Early referral for MV repair before development of atrial fibrillation and LV enlargement may reduce the risk of MR recurrence. Moreover, artificial chordae implantation and ring annuloplasty may assure the long-term durability of MV repair.


An integrated mathematical model of thrombin-, histamine-and VEGF-mediated signalling in endothelial permeability.
Endothelial permeability is involved in injury, inflammation, diabetes and cancer. It is partly regulated by the thrombin-, histamine-, and VEGF-mediated myosin-light-chain (MLC) activation pathways. While these pathways have been investigated, questions such as temporal effects and the dynamics of multi-mediator regulation remain to be fully studied. Mathematical modeling of these pathways facilitates such studies. Based on the published ordinary differential equation models of the pathway components, we developed an integrated model of thrombin-, histamine-, and VEGF-mediated MLC activation pathways. Our model was validated against experimental data for calcium release and thrombin-, histamine-, and VEGF-mediated MLC activation. The simulated effects of PAR-1, Rho GTPase, ROCK, VEGF and VEGFR2 over-expression on MLC activation, and the collective modulation by thrombin and histamine are consistent with experimental findings. Our model was used to predict enhanced MLC activation by CPI-17 over-expression and by synergistic action of thrombin and VEGF at low mediator levels. These may have impact in endothelial permeability and metastasis in cancer patients with blood coagulation. Our model was validated against a number of experimental findings and the observed synergistic effects of low concentrations of thrombin and histamine in mediating the activation of MLC. It can be used to predict the effects of altered pathway components, collective actions of multiple mediators and the potential impact to various diseases. Similar to the published models of other pathways, our model can potentially be used to identify important disease genes through sensitivity analysis of signalling components.


The Extended Treatment Window's Impact on Emergency Systems of Care for Acute Stroke.
The window for acute ischemic stroke treatment was previously limited to 4.5 hours for intravenous tissue plasminogen activator and to 6 hours for thrombectomy. Recent studies using advanced imaging selection expand this window for select patients up to 24 hours from last known well. These studies directly affect emergency stroke management, including prehospital triage and emergency department (ED) management of suspected stroke patients. This narrative review summarizes the data expanding the treatment window for ischemic stroke to 24 hours and discusses these implications on stroke systems of care. It analyzes the implications on prehospital protocols to identify and transfer large-vessel occlusion stroke patients, on issues of distributive justice, and on ED management to provide advanced imaging and access to thrombectomy centers. The creation of high-performing systems of care to manage acute ischemic stroke patients requires academic emergency physician leadership attentive to the rapidly changing science of stroke care.


Epidermal growth factor and trefoil factor family 2 synergistically trigger chemotaxis on BEAS-2B cells via different signaling cascades.
Injured areas of the respiratory epithelium are subject to rapid repair by the migration of adjacent epithelial cells, a process termed "restitution". Rapid re-epithelialization is promoted by interactions between migrating cells and the extracellular matrix proteins. Furthermore, epidermal growth factor (EGF) as well as trefoil factor family (TFF) peptides are well known regulators of epithelial restitution due to their motogenic effects. Migration of the human bronchial epithelial cell line BEAS-2B in modified Boyden chambers was used as a model system for airway restitution. EGF or recombinant human TFF2 or TFF3 showed mainly chemotactic activity. The motogenic response was strictly dependent upon a haptotactic substrate, but to different degrees. EGF induced phosphorylation of extracellular signal-regulated kinases (ERK) 1/2, c-Jun-N-terminal kinase, p38, Akt, and p70S6K in BEAS-2B cells. Using specific inhibitors, the signaling cascades responsible for the motogenic response were shown to differ drastically when EGF was compared with TFF2. The motogenic effect of TFF2 was previously demonstrated to depend on ERK1/2 and protein kinase C activation; whereas the EGF-triggered motogenic response was completely independent of ERK1/2 activation but sensitive to the inhibition of phosphoinositide 3-kinase, p38, protein kinase C, or nuclear factor kappaB. However, the motogenic effects of EGF and TFF2 are additive. These data suggest that luminal EGF and TFF peptides can act synergistically in the human respiratory epithelium to enhance rapid repair processes in the course of diseases such as asthma.


Port malposition in the azygos vein resulting in a veno-broncho and broncho-esophageal fistula: A case report.
We present a case of a port malposition into the azygos vein resulting in both a broncho-esophageal and veno-bronchial fistula. While complications of central venous catheter malposition into the azygos vein are well documented in literature, these unique complications have not yet been described. This case underscores how utilizing state of the art technology like intra-cavity electrocardiography rather that reliance on fluoroscopy can help eliminate catheter malposition and its potential catastrophic consequences.


Isolation and Ambident Reactivity of a Chlorogermylenoid.
Treatment of 2,5-di(3,5-tert-butylphenyl)-1-lithioferrocene with GeCl2 ⋅dioxane afforded the corresponding chlorogermylenoid that exhibited an ambident reactivity in different solvents; it displayed a behavior characteristic for a dichlorogermylene anion in THF, while it exhibited the typical reactivity of a chlorogermylene in toluene. X-Ray diffraction analysis of a single crystal of this chlorogermylenoid, obtained from recrystallization in THF, revealed a separated-ion-pair structure in the solid state.


Assessment of factors affecting practice towards COVID-19 among health care workers in health care facility of West Guji Zone, South Ethiopia, 2020.
health care workers are at greatest risk to being infected with COVID-19 in health care facilities. This study is focused on assessing the level of practice and factors affecting practice towards COVID-19 among health care workers in health care facility of West Guji Zone, Oromia region, Ethiopia. health facility based cross-sectional study design was carried out from December 1st to 30th 2020 among health care providers in West Guji Zone. The simple random sampling technique was used in study and total sample size for this study was 281. The data enter into Epi Data version 188.8.131.52 and SPSS Version 25 used for analysis. The descriptive statistics and logistic regression are needed. The cut point for statically significance settled at p < 0.05. the response rate in this study was 97.8%. Of all study participants, 50.2%, 8.4%, and 6.5% had traveling history, chronic illnesses, and contact with COVID-19 confirmed cases. Too much working, lack of training, and shortage of protective equipment were reported by 54.5%, 50.9%, and 29.1% respectively. About 36.4% and 38.2% of health care providers had poor level of knowledge and prevention practice towards COVID-19. Working at hospital (AOR= 0.156, 95% CI=0.033-0.741), awareness of the action during suggestive symptoms and signs of COVID-19 developed (AOR= 0.038, 95% CI=0.002-0.817), hand washing (AOR= 0.043, 95% CI=0.008-0.238), not going to crowded place (AOR= 0.001, 95% CI=0.001-0.030), applying physical distance (AOR=0.091,95% CI=0.041-0.579) adherence to triage and isolation protocol (AOR=0.317,95%=0.039-0.577)and knowledge level of COVID-19 (AOR=2.378,95%CI=1.523-6.322) were factors significantly associated with prevention practice of COVID-19. in this study, the knowledge level and prevention practice gap was identified. Type of facility, awareness of the action during suggestive symptoms and signs of COVID-19 developed, hands washing to the standard, not going to crowded place, keeping physical distance, adherence to triage and isolation protocol and having good level of knowledge about COVID-19 were factors associated with good prevention practices. Adequate supply of personal protective materials; provision of continuous on-job training and guideline for prevention of COVID-19 must be given to all health care facilities.


[Prognostic value of QKD interval corrected by QRS duration in hypertensive patients].
The QKD interval is measured between the onset of QRS on the ECG and detection of last Korotkoff sound by a microphone placed on the brachial artery while measuring BP. It is the sum of preejection time and pulse transmission time and thus is correlated to pulse wave velocity (PWV). This interval is automatically monitored with BP and HR every 15 minutes during 24 h with an ambulatory blood pressure monitoring device (Diasys integra, Novacor, France). The 96 measurements obtained allow to automatically calculate the QKD(100-60), QKD value for 100 mmHg SBP and 60 bpm HR. This indice of arterial stiffness has been shown to be linked to future cardiovascular (CV) events, independently of 24 h BP. However this interval may be abnormally prolonged in case of left bundle branch blocks (LBBB). We tested the effects of simply removing QRS duration from QKD(100-60) value on the prediction of CV events in a population of 412 hypertensives (247 males: age = 53 +/- 14 years; office BP = 158 +/- 19/97 +/- 11 mmHg; 24 h BP = 133 +/- 17/86 +/- 11 mmHg) followed prospectively. Mean follow up was 65 months, 32 pts were lost, 49 CV events occurred including 11 deaths. Cox model showed that baseline QKD(100-60) (m = 202 +/- 19; 142-254 ms) was significantly (p < 0.05) associated to events independently of age, 24h SBP and other traditional risk factors. Removing QRS duration (m = 85 +/- 10: 61-158 ms) improves the relation to events (monovariate khi2 = 38 vs 30). Removing QRS duration from QKD(100-60) improves its predictive value of future CV events and allows using this method in patients with LBBB.


Feasibility of administering aminoglycoside antibiotics by continuous intravenous infusion.
Eleven patients each received gentamicin sulfate, tobramycin, and sisomicin by continuous intravenous infusion after an initial loading dose. Subsequent doses of antibiotic were adjusted in an attempt to maintain constant serum concentrations. There was considerable variation in the serum concentration from patient to patient and in the same patient from day to day with each drug. Although the dosages of gentamicin sulfate and tobramycin were similar, the serum concentrations of the latter drug were consistently lower. Despite the daily administration of doses of at least 300 mg of gentamicin and tobramycin per m(2) and 160 mg of sisomicin per m(2), nephrotoxicity occurred in only three patients. This is a low frequency of nephrotoxicity, considering the dosages of drug administered. Although therapeutic efficacy was not an objective of this study, 8 of 11 documented infections were cured. This approach to the administration of aminoglycoside antibiotics deserves therapeutic trials.


Role of quantum coherence in shaping the line shape of an exciton interacting with a spatially and temporally correlated bath.
Kubo's fluctuation theory of line shape forms the backbone of our understanding of optical and vibrational line shapes, through such concepts as static heterogeneity and motional narrowing. However, the theory does not properly address the effects of quantum coherences on optical line shape, especially in extended systems where a large number of eigenstates are present. In this work, we study the line shape of an exciton in a one-dimensional lattice consisting of regularly placed and equally separated optical two level systems. We consider both linear array and cyclic ring systems of different sizes. Detailed analytical calculations of line shape have been carried out by using Kubo's stochastic Liouville equation (SLE). We make use of the observation that in the site representation, the Hamiltonian of our system with constant off-diagonal coupling J is a tridiagonal Toeplitz matrix (TDTM) whose eigenvalues and eigenfunctions are known analytically. This identification is particularly useful for long chains where the eigenvalues of TDTM help understanding crossover between static and fast modulation limits. We summarize the new results as follows. (i) In the slow modulation limit when the bath correlation time is large, the effects of spatial correlation are not negligible. Here the line shape is broadened and the number of peaks increases beyond the ones obtained from TDTM (constant off-diagonal coupling element J and no fluctuation). (ii) However, in the fast modulation limit when the bath correlation time is small, the spatial correlation is less important. In this limit, the line shape shows motional narrowing with peaks at the values predicted by TDTM (constant J and no fluctuation). (iii) Importantly, we find that the line shape can capture that quantum coherence affects in the two limits differently. (iv) In addition to linear chains of two level systems, we also consider a cyclic tetramer. The cyclic polymers can be designed for experimental verification. (v) We also build a connection between line shape and population transfer dynamics. In the fast modulation limit, both the line shape and the population relaxation, for both correlated and uncorrelated bath, show similar behavior. However, in slow modulation limit, they show profoundly different behavior. (vi) This study explains the unique role of the rate of fluctuation (inverse of the bath correlation time) in the sustenance and propagation of coherence. We also examine the effects of off-diagonal fluctuation in spectral line shape. Finally, we use Tanimura-Kubo formalism to derive a set of coupled equations to include temperature effects (partly neglected in the SLE employed here) and effects of vibrational mode in energy transfer dynamics.


Diagnostic Assessment Reviews: is cost-effectiveness analysis helpful or necessary?
Objective Diagnostic Assessment Reviews are part of the work programme of the National Institute of Health and Care Excellence (NICE) in England to evaluate emergent medical technologies and so ensure timely and consistent adoption within the National Health Service (NHS). New diagnostics have the potential to improve health outcomes and impact positively on health service resource use. This study sought to assess the quality of economic evidence informing Diagnostic Assessment Reviews. Methods We reviewed 27 Diagnostic Assessment Reviews that had been published by NICE as of 30 May 2017 by summarizing and interpreting the evidence that was used to carry out cost-effectiveness analyses. Common issues and challenges of the assessment process were illustrated. DARs differed in the methods and assumptions used to construct economic models, and linkage of economic model and diagnostic findings. Even though some diagnostic technologies were estimated to be cost-effective, they were not always adopted for routine use in the NHS. The majority of Diagnostic Assessment Review economic models relied heavily on assumptions and expert opinion, with considerable uncertainty about the cost-effectiveness of diagnostic testing. Conclusions Diagnostic Assessment Reviews appraisals as commissioned by NICE typically feature varying evidence for diagnostic performance and limited evidence for resource implications and quality of life, often leading to recommendations for further research. Given the process opportunity cost, a two-stage topic selection process, with initial assessment specifying further research and proceeding to full assessment when adequate evidence is available may be more appropriate and help signal to diagnostics companies the type of research evidence required.


The role of soluble CD23 in distinguishing stable and progressive forms of B-chronic lymphocytic leukemia.
Soluble CD23 (sCD23) has been recognized as an important prognostic parameter in patients with chronic lymphocytic leukemia (B-CLL) at early clinical stages. There is, however, no clear information on its prognostic significance in advanced stages and on its role as an indicator for aggressive or indolent courses of disease. Therefore, sCD23 was measured in the serum of 145 patients at diagnosis and serial determinations were carried out for 8 years in 38 patients. The results indicate that in patients with identical clinical stages at first presentation the disease could take different courses depending on initial sCD23 concentrations below or above specific threshold levels (860 and 5900U/ml). sCD23 higher than these thresholds was associated with faster progression into upper clinical stages. Furthermore, sCD23-doubling time (sCD23-DT) indicated that patients with long DT progressed slowly, while those with short DT had more aggressive disease. Particularly in patients with advanced disease stages, long sCD23-DT indicated development of smoldering disease. Since sCD23 levels reflect total tumor mass, determination of sCD23-DT has probably a better predictive value than lymphocyte doubling time. It appears that B-CLL patients can be divided into different risk categories according to initial determinations of sCD23 and that sCD23-DT is an additional important parameter in predicting disease progression.


Immune Response to Extracellular Vesicles From Human Islets of Langerhans in Patients With Type 1 Diabetes.
The autoimmune response that characterizes type 1 diabetes (T1D) has no clear cause. Extracellular vesicles (EVs) play an important role in triggering the immune response in other contexts. Here, we propose a model by which EVs isolated from human islets stimulate proinflammatory immune responses and lead to peripheral blood mononuclear cell (PBMC) activation. We show that human islet EVs are internalized by monocytes and B cells and lead to an increase in T-helper 1, 2, and 17 cytokine expression, as well as T and B cell proliferation. Importantly, we demonstrate memory T and B cell activation by EVs selectively in PBMCs of patients with T1D. Additionally, human islet EVs induce an increase in antibodies against glutamic acid decarboxylase 65 (GAD65) in T1D PBMCs. Furthermore, pretreatment of T1D PBMCs with ibrutinib, an inhibitor of Bruton tyrosine kinase, dampens EV-induced memory B cell activation and GAD65 antibody production. Collectively, our findings indicate a role for human islet EVs in mediating activation of B and T cells and GAD65 autoantibody production.


Gamification to Improve Medication Adherence: A Mixed-method Usability Study for MedScrab.
Medication non-adherence is a prevalent healthcare problem with poor health outcomes and added healthcare costs. MedScrab, a gamification-based mHealth app, is the first attempt to deliver crucial life-saving medication information to patients and increase their medication adherence. The paper presents the development of MedScrab and a two-phase mixed-method usability evaluation of MedScrab. Phase I qualitatively evaluated MedScrab using a think-aloud protocol for its usability. With 51 participants, qualitative data analysis of Phase I revealed two themes: positive functionality of the app and four areas of improvement. The improvement recommendations were incorporated into MedScrab's design. Phase I also validated a widely used mHealth App Usability Questionnaire (MAUQ). Quantitative data analysis of Phase I reduced the original 18-item MAUQ scale to a 15-item scale with two factors: ease of use (4 items) and usefulness and satisfaction (11 items). Phase II surveyed 83 participants from Amazon's Mechanical Turk using a modified MAUQ. The modified MAUQ scale showed strong internal consistency (Cronbach alpha = 0.959) and high factor loadings (between 0.623 and 0.987). The study design of the usability evaluation can serve as a methodological guide for designing, evaluating, and improving mHealth apps.The usability study showed that MedScrab was perceived as ease of use (6.24 out of 7) with high usefulness and satisfaction (5.72 out of 7). The quantitative data analysis results support the use of the modified MAUQ as a valid instrument to measure the usability of the MedScrab. However, the instrument should be used with adaptation based on the app's characteristics.


Fast-Clearable Nanocarriers Conducting Chemo/Photothermal Combination Therapy to Inhibit Recurrence of Malignant Tumors.
Inhomogeneous heating by photothermal therapy (PTT) during cancer treatment often results in the recurrence of tumors. Thus, integrating PTT with chemotherapy (CHT) may provide a complementary treatment for enhanced therapeutic efficiency. Herein, this study develops a hollow structured polymer-silica nanohybrid (HPSN) as a nanocarrier to simultaneously deliver the anticancer drug paclitaxel and photothermal agent palladium phthalocyanine to tumors through enhanced permeation and the retention effect. A combinational CHT/PTT therapy on mice bearing aggressive tumor grafts is conducted. The highly malignant tumor model, which recurs after sole treatment of PTT, can be eradicated by the combined CHT/PTT treatment. In addition, most of the off-targeted HPSN nanocarriers can be excreted through a hepatobiliary pathway in about 10 d. Serology results show that the fast-clearable HPSN can significantly reduce the side effect of the loaded paclitaxel drug. The present work provides an alternative approach for combinational cancer treatment with high therapeutic efficiency.


A glyoxalase I inhibitor of a new structural type produced by Streptomyces.
Many streptomyces strains produced an inhibitor of crude glyoxalase prepared from rat liver which did not inhibit glyoxalase I prepared from yeast. Another inhibitor, C11H14O6, which inhibited glyoxalases prepared from both rat liver and yeast was obtained from a cultured broth of Streptomyces griseosproeus and crystallized. Preincubation of this inhibitor with reuduced glutathione increased its inhibitory activity, which suggested its reaction with reduced glutathione. It showed a strong inhibition of growth of HeLa cells and inhibition of Ehrlich ascites carcinoma by daily injection. It also showed weak inhibition of the solid type of Ehrlich carcinoma and prolonged the survival period of mice inoculated with L-1210 cells.


The effects of pulsed auditory stimulation on various gait measurements in persons with Parkinson's Disease.
The purpose of this study was to examine the Functional Ambulation Performance Score (FAP; a quantitative gait measure) in persons with Parkinson's Disease (PD) using the auditory stimulation of a metronome (ASM). Participants (n = 16; 5F/11M; range 60--84 yrs.) had a primary diagnosis of PD and were all independent ambulators. Footfall data were collected while participants walked multiple times on an electronic walkway under the following conditions: 1) PRETEST: establishing baseline cadence, 2) ASM: metronome set to baseline cadence, 3) 10ASM: metronome set to 10% FAP scores increased between PRETEST and POSTTEST. PRE/POSTTEST comparisons also indicated decreases in cycle time and double support and increases in step length and step-extremity ratio (step length/leg length). The results confirm prior findings that auditory stimulation can be used to positively influence the gait of persons with PD and suggest beneficial effects of ASM as an adjunct to dopaminergic therapy to treat gait dysfunctions in PD.


A cell cycle-independent, conditional gene inactivation strategy for differentially tagging wild-type and mutant cells.
Here, we describe a novel method based on intronic MiMIC insertions described in Nagarkar-Jaiswal et al. (2015) to perform conditional gene inactivation in Drosophila. Mosaic analysis in Drosophila cannot be easily performed in post-mitotic cells. We therefore, therefore, developed Flip-Flop, a flippase-dependent in vivo cassette-inversion method that marks wild-type cells with the endogenous EGFP-tagged protein, whereas mutant cells are marked with mCherry upon inversion. We document the ease and usefulness of this strategy in differential tagging of wild-type and mutant cells in mosaics. We use this approach to phenotypically characterize the loss of SNF4Aγ, encoding the γ subunit of the AMP Kinase complex. The Flip-Flop method is efficient and reliable, and permits conditional gene inactivation based on both spatial and temporal cues, in a cell cycle-, and developmental stage-independent fashion, creating a platform for systematic screens of gene function in developing and adult flies with unprecedented detail.


Neoplasia de celulas dendriticasblastica plasmocitoide.
Plasmacytoid blast dendritic cell neoplasia (NCDBP) is an uncommon malignant neoplasm, presenting clinically with cutaneous involvement and subsequent lymph node and bone marrow extension. It characteristically expresses the markers: CD56, CD4, and CD123. There is no optimal treatment, relapses are frequent, and the survival time is short. We present the case of an elderly patient with NCDBP who initially presented with cutaneous lesions, but experienced rapid systemic progression and did not, respond to treatment.


Effect of N-acetylcysteine on the pulmonary response to endotoxin in the awake sheep and upon in vitro granulocyte function.
Oxygen free radicals released during endotoxemia may contribute to the lung injury of the adult respiratory distress syndrome (ARDS). As this syndrome occurs frequently after gram-negative sepsis in humans, we studied the effect of intravenous N-acetylcysteine (NAC), a free radical scavenger, upon the endotoxin (E)-induced model of ARDS in awake sheep. In vivo studies demonstrated that NAC attenuates the endotoxin-induced rise in pulmonary artery pressure (62 +/- 3 torr with E control vs. 43 +/- 3 torr for E + NAC), and markedly diminishes the rise in lymph flow at 1 h (8.5 +/- 1.2 vs 4.5 +/- 0.6 ml/15 min) and 4 h (5.0 +/- 0.6 vs. 3.3 +/- 0.4 ml/15 min), respectively, for E control vs. E + NAC. NAC also markedly attenuated the alterations in lung mechanics after endotoxemia. Dynamic compliance at 2 h after endotoxemia was 44 +/- 6% of base line for E vs. 76 +/- 10% of base line for E + NAC. Resistance to airflow across the lung at 1 h postendotoxin was 811 +/- 280% of base line for E vs. 391 +/- 233% of base line for E + NAC. NAC substantially reduced the 1 h postendotoxin rise in lymph concentrations of thromboxane B2 (8.29 +/- 3.28 vs. 2.75 +/- 1.93 ng/ml for E vs. E + NAC) and 6-keto-prostaglandin-F1 alpha (0.91 +/- 0.27 vs. 0.23 +/- 0.12 ng/ml for E vs. E + NAC). In addition, in vitro studies were performed which revealed NAC to be a potent free radical scavenger in both biologic and nonbiologic free radical generating systems. NAC decreased phorbol-stimulated granulocyte aggregation in a concentration-dependent manner in vitro. Minimal effects were observed, however, upon leukocyte degranulation at the concentrations of NAC achieved during the in vivo tests. Thus, NAC significantly attenuated all monitored pathophysiologic changes in the endotoxin model of ARDS in sheep, possibly by its ability to scavenge toxic oxygen free radicals. A direct impairment of the ability of inflammatory cells to generate oxygen radicals cannot be ruled out.


Genetically Encoded Calcium Indicators: A New Tool in Renal Hypertension Research.
Hypertension is ranked as the third cause of disability-adjusted life-years. The percentage of the population suffering from hypertension will continue to increase over the next years. Renovascular disease is one of the most common causes of secondary hypertension. Vascular changes seen in hypertension are partially based on dysfunctional calcium signaling. This signaling can be studied using calcium indicators (loading dyes and genetically encoded calcium indicators; GECIs). Most progress in development has been seen in GECIs, which are used in an increasing number of publications concerning calcium signaling in vasculature and the kidney. The use of transgenic mouse models expressing GECIs will facilitate new possibilities to study dysfunctional calcium signaling in a cell type-specific manner, thus helping to identify more specific targets for treatment of (renal) hypertension.


Decellularization Strategies for Regenerative Medicine: From Processing Techniques to Applications.
As the gap between donors and patients in need of an organ transplant continues to widen, research in regenerative medicine seeks to provide alternative strategies for treatment. One of the most promising techniques for tissue and organ regeneration is decellularization, in which the extracellular matrix (ECM) is isolated from its native cells and genetic material in order to produce a natural scaffold. The ECM, which ideally retains its inherent structural, biochemical, and biomechanical cues, can then be recellularized to produce a functional tissue or organ. While decellularization can be accomplished using chemical and enzymatic, physical, or combinative methods, each strategy has both benefits and drawbacks. The focus of this review is to compare the advantages and disadvantages of these methods in terms of their ability to retain desired ECM characteristics for particular tissues and organs. Additionally, a few applications of constructs engineered using decellularized cell sheets, tissues, and whole organs are discussed.


Nitrite, biogenic amines and volatile N-nitrosamines in commercial Chinese traditional fermented fish products.
To examine the safety of Chinese traditional fermented fish products (CTFPs) available on the Chinese market, nitrite, nitrate, biogenic amines (BAs) and volatile N-nitrosamines (VNAs) content in 33 commercial CTFPs from different provinces was investigated. The mean content of nitrite and nitrate wase 0.63 and 749.5 mg/kg, respectively. Concerning the occurrence of BAs, the accumulation in all CTFPs samples remained at low levels, whereas only in one sample from Guangxi the histamine content exceeded the critical level (50 mg/kg). In addition, six types of VNAs, including N-nitrosodimethylamine (NDMA), N-nitrosoethylmethylamine, N-nitrosopiperidine, N-nitrosopyrrolidine, N-nitrosomorpholine and N-nitrosodiphenylamine, were detected in a high number of samples. The NDMA content in 36.4% of the samples and the total VNAs content in about 63.6% of the samples were unacceptable. Principal component analysis indicated that the accumulation of NDMA and total VNAs was closely related with the content of histamine, tyramine and nitrate.


Structural and functional alterations in Malpighian tubules as biomarkers of environmental pollution: synopsis and prospective.
Although a number of biomarkers of pollutant exposure have been identified in invertebrate species, little is known about the effect on Malpighian tubules playing an essential role in excretion and osmoregulation. Analyses of structural and functional alterations on this organ can be useful to predict the effects at the organism and population level in monitoring studies of environmental pollution. The aim of the present review is to provide a synthesis of existing knowledge on cellular damages induced by xenobiotics in Malpighian tubules both under laboratory and field conditions. We compared studies of exposure to pesticides and heavy metals as mainly environmental contaminants from anthropogenic activities. This report provided evidence that the exposure to xenobiotics has an effect on this organ and reinforces the need for further research integrating molecular biomarkers with analysis on Malpighian tubules. Copyright © 2017 John Wiley & Sons, Ltd.


Effectiveness of a multimodal intervention to improve blood culture collection in an adult emergency department.
We investigated the impact of a multimodal intervention to improve the compliance of BC collections as a composite outcome, taking into account both blood volume collected and absence of solitary BC. We performed a quasi-experimental study using a before-after design (5 months for pre- and post-intervention evaluation) in an adult emergency department at a tertiary care hospital that showed that a multimodal intervention was associated with a dramatic increase in the proportion of blood cultures that were collected as recommended per national guidelines, from 17.3% (328/1896) to 68.9% (744/1080), P < 0.0001. The implementation of such intervention in other settings could improve the diagnosis of bloodstream infections and reduce irrelevant costs.


Controls on eDNA movement in streams: Transport, Retention, and Resuspension.
Advances in detection of genetic material from species in aquatic ecosystems, including environmental DNA (eDNA), have improved species monitoring and management. eDNA from target species can readily move in streams and rivers and the goal is to measure it, and with that infer where and how abundant species are, adding great value to delimiting species invasions, monitoring and protecting rare species, and estimating biodiversity. To date, we lack an integrated framework that identifies environmental factors that control eDNA movement in realistic, complex, and heterogeneous flowing waters. To this end, using an empirical approach and a simple conceptual model, we propose a framework of how eDNA is transported, retained, and resuspended in stream systems. Such an understanding of eDNA dispersal in streams will be essential for designing optimized sampling protocols and subsequently estimating biomass or organismal abundance. We also discuss guiding principles for more effective use of eDNA methods, highlighting the necessity of understanding these parameters for use in future predictive modeling of eDNA transport.


Significance of ammonia in growth of atmospheric nanoclusters.
We apply accurate quantum chemistry methods to study the thermochemistry of molecular clusters containing ammonia, water, and sulfuric acid and investigate initial reaction steps in atmospheric nucleation by calculating free energies for the related reactions. The results indicate that ammonia is a key reactant enhancing the growth of small water-sulfuric acid clusters in atmospheric conditions. The role of ammonia becomes significant when the nanoclusters contain more than one or two sulfuric acid molecules. This implies a lower limit of 1:3 for the NH3/H2SO4 mole ratio of atmospheric sulfuric acid-water-ammonia clusters.


Effects of lidocaine on myocardial contractility and baroreflex control of heart rate in conscious dogs.
The effects of intravenous lidocaine (30 and 60 micrograms X kg-1 X min-1 during 30 min) at steady-state plasma levels (1.9 +/- 0.2 and 3.5 +/- 0.2 micrograms/ml, respectively) were investigated in conscious dogs, previously instrumented for measurements of arterial and left ventricular (LV) pressures, isometric myocardial contractility indexes (LV peak rate of tension development [dP/dt] and LV [dP/dt]/DP40), and heart rate. In addition, before and at the end of lidocaine infusions, arterial baroreflex responses were tested by bolus injections of nitroglycerin and phenylephrine. Whereas LV peak dP/dt and LV (dP/dt)/DP40 were significantly decreased after the low dosage of lidocaine, these indexes returned to control values after the 10th min of infusion of the high dosage. Moreover, eight out of 14 dogs exhibited continuous tremors, tachycardia, hypertension, and increase in contractility after the 10th min of lidocaine infusion (60 micrograms X kg-1 X min-1), although their lidocaine plasma levels (3.7 +/- 0.2 micrograms/ml) did not differ from those of the whole group. When these dogs were pretreated by combined alpha- and beta-adrenoceptor blocking drugs, none of them had tremors, and there was a constant depressant effect on cardiac chronotropism and inotropism. A specific enhancement of baroreflex sensitivity after phenylephrine injection was observed at the high lidocaine dosage. It is concluded that a central stimulation of both components of the autonomic nervous system modulates the direct effects of therapeutic plasma levels of lidocaine on cardiac chronotropism and inotropism in conscious dogs.


Immune response to uncoupled peptides of foot-and-mouth disease virus.
Uncoupled synthetic peptide representing the sequence of amino acids 141-160 of foot-and-mouth disease virus (FMDV) protein VP1 induced a virus-neutralizing antibody response in guinea-pigs. This response required incomplete Freund's adjuvant (IFA) for the primary inoculation and was dependent on the presence of an added cysteine residue with an unblocked sulphydryl group at the carboxy-terminus. Secondary immunization could be carried out in the absence of adjuvant. A study of the relative activities of nested sets of uncoupled peptides from 150-160 to 135-160 and 141-160 to 141-155 indicated that amino acids 146-156 were critical for the induction of virus-neutralizing antibodies and that extension to 137-160 further improved this response. Results of in vitro proliferation studies demonstrated that the carboxy-terminal residues on this peptide may form a T-cell epitope. The significance of these observations in the broader context of synthetic peptide vaccines is discussed.


Early psychotherapeutic empathy, alliance, and client outcome: Preliminary evidence of indirect effects.
The association between psychotherapeutic empathy and client outcome is well established, yet the mechanisms underlying this association remain poorly understood. We hypothesized that early experiences of empathy influence outcome through the working alliance. To test this hypothesis, we used archival data collected from 56 clients (mean [M] age = 19.5 years, 83.9% female, 76.8% White) who reported mild, moderate, or severe depressive symptoms at screening and pretreatment assessments and then received five sessions of evidence-based psychotherapy. Therapists (M age = 26.0 years, 50% female, 100% White) were six students in a clinical psychology PhD program. Results of bootstrap analyses were consistent with the idea that early experiences of empathy strengthen the alliance (specifically the goals and tasks facets), which in turn facilitates improvements in depressive symptoms and psychological well-being. While preliminary, these results implicate a specific pathway of change in the treatment of depression.


Thyroid disease and lipids.
The composition and the transport of lipoproteins are seriously disturbed in thyroid diseases. Overt hypothyroidism is characterized by hypercholesterolaemia and a marked increase in low-density lipoproteins (LDL) and apolipoprotein B (apo A) because of a decreased fractional clearance of LDL by a reduced number of LDL receptors in the liver. The high-density lipoprotein (HDL) levels are normal or even elevated in severe hypothyroidism because of decreased activity of cholesteryl-ester transfer protein (CETP) and hepatic lipase (HL), which are enzymes regulated by thyroid hormones. The low activity of CETP, and more specifically of HL, results in reduced transport of cholesteryl esters from HDL(2) to very low-density lipoproteins (VLDL) and intermediate low-density lipoprotein (IDL), and reduced transport of HDL(2) to HDL(3). Moreover, hypothyroidism increases the oxidation of plasma cholesterol mainly because of an altered pattern of binding and to the increased levels of cholesterol, which presents a substrate for the oxidative stress. Cardiac oxygen consumption is reduced in hypothyroidism. This reduction is associated with increased peripheral resistance and reduced contractility. Hypothyroidism is often accompanied by diastolic hypertension that, in conjunction with the dyslipidemia, may promote atherosclerosis. However, thyroxine therapy, in a thyrotropin (TSH)-suppressive dose, usually leads to a considerable improvement of the lipid profile. The changes in lipoproteins are correlated with changes in free thyroxine (FT(4)) levels. Hyperthyroidism exhibits an enhanced excretion of cholesterol and an increased turnover of LDL resulting in a decrease of total and LDL cholesterol, whereas HDL are decreased or not affected. The action of thyroid hormone on Lp(a) lipoprotein is still debated, because both decrease or no changes have been reported. The discrepancies are mostly because of genetic polymorphism of apo(a) and to the differences between the various study groups. Subclinical hypothyroidism (SH) is associated with lipid disorders that are characterized by normal or slightly elevated total cholesterol levels, increased LDL, and lower HDL. Moreover, SH has been associated with endothelium dysfunction, aortic atherosclerosis, and myocardial infarction. Lipid disorders exhibit great individual variability. Nevertheless, they might be a link, although it has not been proved, between SH and atherosclerosis.


Two orders of magnitude enhancement in oxygen evolution reactivity on amorphous Ba0.5Sr0.5Co0.8Fe0.2O3-δ nanofilms with tunable oxidation state.
Perovskite oxides exhibit potential for use as electrocatalysts in the oxygen evolution reaction (OER). However, their low specific surface area is the main obstacle to realizing a high mass-specific activity that is required to be competitive against the state-of-the-art precious metal-based catalysts. We report the enhanced performance of Ba0.5Sr0.5Co0.8Fe0.2O3-δ (BSCF) for the OER with intrinsic activity that is significantly higher than that of the benchmark IrO2, and this result was achieved via fabrication of an amorphous BSCF nanofilm on a surface-oxidized nickel substrate by magnetron sputtering. The surface nickel oxide layer of the Ni substrate and the thickness of the BSCF film were further used to tune the intrinsic OER activity and stability of the BSCF catalyst by optimizing the electronic configuration of the transition metal cations in BSCF via the interaction between the nanofilm and the surface nickel oxide, which enables up to 315-fold enhanced mass-specific activity compared to the crystalline BSCF bulk phase. Moreover, the amorphous BSCF-Ni foam anode coupled with the Pt-Ni foam cathode demonstrated an attractive small overpotential of 0.34 V at 10 mA cm-2 for water electrolysis, with a BSCF loading as low as 154.8 μg cm-2.


Successful validation of genomic biomarkers for human immunotoxicity in Jurkat T cells in vitro.
Previously, we identified 25 classifier genes that were able to assess immunotoxicity using human Jurkat T cells. The present study aimed to validate these classifiers. For that purpose, Jurkat cells were exposed for 6 h to subcytotoxic doses of nine immunotoxicants, five non-immunotoxicants and four compounds for which human immunotoxicity has not yet been fully established. RNA was isolated and subjected to Fluidigm quantitative real time (qRT)-PCR analysis. The sensitivity, specificity and accuracy of the screening assay as based on the nine immunotoxicants and five non-immunotoxicants used in this study were 100%, 80% and 93%, respectively, which is better than the performance in our previous study. Only one compound was classified as false positive (benzo-e-pyrene). Of the four potential (non-)immunotoxicants, chlorantraniliprole and Hidrasec were classified immunotoxic and Sunset yellow and imidacloprid as non-immunotoxic. ToxPi analysis of the PCR data provided insight in the molecular pathways that were affected by the compounds. The immunotoxicants 2,3-dichloro-propanol and cypermethrin, although structurally different, affected protein metabolism and cholesterol biosynthesis and transport. In addition, four compounds, i.e. chlorpyrifos, aldicarb, benzo-e-pyrene and anti-CD3, affected genes in cholesterol metabolism and transport, protein metabolism and transcription regulation. qRT-PCR on eight additional genes coding for similar processes as defined in ToxPi analyzes, supported these results. In conclusion, the 25 immunotoxic classifiers performed very well in a screening with new non-immunotoxic and immunotoxic compounds. Therefore, the Jurkat screening assay has great promise to be applied within a tiered approach for animal free testing of human immunotoxicity.


[Correlation of FGF2 and TGF-β1 with benign prostatic hyperplasia: Progress in research].
Benign prostatic hyperplasia (BPH) is a most common disorder in the elderly. The development and growth of the prostate are regulated by androgens and growth factors, and the latter are secreted by prostatic epithelial and mesenchymal cells and play a key role. As the family members of FGF and TGF-β, FGF2 and TGF-β1 are the most representative growth factors in the prostate, involved in the regulation of such biological processes as proliferation, apoptosis, migration and differentiation of prostate cells, which provides a theoretical basis for the clinical application of FGF- and TGF-β-targeted drugs in the treatment of BPH.


Surface salt bridges contribute to the extreme thermal stability of an FN3-like domain from a thermophilic bacterium.
This study uses differential scanning calorimetry, X-ray crystallography, and molecular dynamics simulations to investigate the structural basis for the high thermal stability (melting temperature 97.5°C) of a FN3-like protein domain from thermophilic bacteria Thermoanaerobacter tengcongensis (FN3tt). FN3tt adopts a typical FN3 fold with a three-stranded beta sheet packing against a four-stranded beta sheet. We identified three solvent exposed arginine residues (R23, R25, and R72), which stabilize the protein through salt bridge interactions with glutamic acid residues on adjacent strands. Alanine mutation of the three arginine residues reduced melting temperature by up to 22°C. Crystal structures of the wild type (WT) and a thermally destabilized (∆Tm -19.7°C) triple mutant (R23L/R25T/R72I) were found to be nearly identical, suggesting that the destabilization is due to interactions of the arginine residues. Molecular dynamics simulations showed that the salt bridge interactions in the WT were stable and provided a dynamical explanation for the cooperativity observed between R23 and R25 based on calorimetry measurements. In addition, folding free energy changes computed using free energy perturbation molecular dynamics simulations showed high correlation with melting temperature changes. This work is another example of surface salt bridges contributing to the enhanced thermal stability of thermophilic proteins. The molecular dynamics simulation methods employed in this study may be broadly useful for in silico surface charge engineering of proteins.