Three-Dimensional Domain Swapping Changes the Folding Mechanism of the Forkhead Domain of FoxP1.
The forkhead family of transcription factors (Fox) controls gene transcription during key processes such as regulation of metabolism, embryogenesis, and immunity. Structurally, Fox proteins feature a conserved DNA-binding domain known as forkhead. Interestingly, solved forkhead structures of members from the P subfamily (FoxP) show that they can oligomerize by three-dimensional domain swapping, whereby structural elements are exchanged between adjacent subunits, leading to an intertwined dimer. Recent evidence has largely stressed the biological relevance of domain swapping in FoxP, as several disease-causing mutations have been related to impairment of this process. Here, we explore the equilibrium folding and binding mechanism of the forkhead domain of wild-type FoxP1, and of two mutants that hinder DNA-binding (R53H) and domain swapping (A39P), using size-exclusion chromatography, circular dichroism, and hydrogen-deuterium exchange mass spectrometry. Our results show that domain swapping of FoxP1 occurs at micromolar protein concentrations within hours of incubation and is energetically favored, in contrast to classical domain-swapping proteins. Also, DNA-binding mutations do not significantly affect domain swapping. Remarkably, equilibrium unfolding of dimeric FoxP1 follows a three-state N2 ↔ 2I ↔ 2U folding mechanism in which dimer dissociation into a monomeric intermediate precedes protein unfolding, in contrast to the typical two-state model described for most domain-swapping proteins, whereas the A39P mutant follows a two-state N ↔ U folding mechanism consistent with the second transition observed for dimeric FoxP1. Also, the free-energy change of the N ↔ U in A39P FoxP1 is ∼2 kcal⋅mol(-1) larger than the I ↔ U transition of both wild-type and R53H FoxP1. Finally, hydrogen-deuterium exchange mass spectrometry reveals that the intermediate strongly resembles the native state. Our results suggest that domain swapping in FoxP1 is at least partially linked to monomer folding stability and follows an unusual three-state folding mechanism, which might proceed via transient structural changes rather than requiring complete protein unfolding as do most domain-swapping proteins.


The role of collagen in bone structure: an image processing approach.
Bone collagen structure in normal and pathological tissues is illustrated using techniques of thin section transmission electron microscopy and computer-assisted analysis. The normal bone collagen types, fibril architecture and diameter are described. In pathological tissue, deviations from normal fine structure are reflected in abnormal arrangements of collagen fibrils and abnormalities in fibril diameter. Computer analyses of normal fibril positive staining patterns are presented in order to provide a basis for comparing such patterns with pathological ones.


Mechanisms Responsible for ω-Pore Currents in Cav Calcium Channel Voltage-Sensing Domains.
Mutations of positively charged amino acids in the S4 transmembrane segment of a voltage-gated ion channel form ion-conducting pathways through the voltage-sensing domain, named ω-current. Here, we used structure modeling and MD simulations to predict pathogenic ω-currents in CaV1.1 and CaV1.3 Ca2+ channels bearing several S4 charge mutations. Our modeling predicts that mutations of CaV1.1-R1 (R528H/G, R897S) or CaV1.1-R2 (R900S, R1239H) linked to hypokalemic periodic paralysis type 1 and of CaV1.3-R3 (R990H) identified in aldosterone-producing adenomas conducts ω-currents in resting state, but not during voltage-sensing domain activation. The mechanism responsible for the ω-current and its amplitude depend on the number of charges in S4, the position of the mutated S4 charge and countercharges, and the nature of the replacing amino acid. Functional characterization validates the modeling prediction showing that CaV1.3-R990H channels conduct ω-currents at hyperpolarizing potentials, but not upon membrane depolarization compared with wild-type channels.


Seed fall and phenological overlap in a guild of ant-dispersed herbs.
The timing of seed fall was studied in a guild of seven ant-dispersed forest herbs in NE-Zealand, Denmark. It is demonstrated, that the seed fall curves of Carex pilulifera, Melica uniflora and Luzula multiflora are remarkably different, representing logistic, hyperbolic, and linear relationships, respectively. The influence of environmental factors, and the significance to dispersal and predation, is discussed. It is suggested, that the three types of curves represent different strategies of seed fall. Simple, and explanded models of seed fall rate indicate that the density of seeds available to ants was more than 20 times greater for C. pilulifera than for M. uniflora. The observed phenological overlap of dispersal periods in the guild conformed to random expectations. Thus, the competition hypothesis was rejected, as there were no indications of a shift in phenology resulting from interspecific competition for the dispersal vector.


Hemostatic changes by thrombopoietin-receptor agonists in immune thrombocytopenia patients.
Thrombopoietin receptor agonist (TPO-RA) treatment increases the thrombosis rate in immune thrombocytopenia (ITP). We hypothesize that TPO-RAs influence platelet function, global and secondary hemostasis and/or fibrinolysis. A systematic review was performed. If possible, data were compared between responders (relevant increase in platelet count), and non-responders. Twelve observational studies with 305 patients were included (responders (127/150 (85%))). There were indications that TPO-RA treatment enhanced platelet function, with respect to platelet-monocyte aggregates, soluble P-selectin, GPVI expression, and adhesion under flow. Studies addressing global and secondary hemostasis and fibrinolysis were scarce. Overall, no changes were found during TPO-RA treatment, apart from an accelerated clot formation and conflicting data on levels of plasminogen activator inhibitor (PAI)-1. The parameters that increased have previously been associated with thrombosis in other patient groups, and might contribute to the increased rate of thrombosis observed in TPO-RA-treated ITP patients.


Transport of L-leucine hydroxy analogue and L-lactate in human small intestinal brush border membrane vesicles.
Nitrogen load can be reduced by substituting dietary protein intake with keto or hydroxy analogues of amino acids. To investigate their intestinal absorption, human jejunal brush border membrane vesicles were used to measure uptake of L-leucine hydroxy analogue (L-LHA) and L-lactate. Uptake assays were performed under voltage-clamped conditions in the presence of valinomycin and K+ inside and outside. Both inward directed H(+)- and Na(+)-gradients stimulated uptake of both substrates. The H(+)-gradient was the major driving force and led to an increased rate of L-LHA or L-lactate transport. The proton ionophore FCCP abolished the H(+)-gradient-driven but not the Na(+)-gradient-driven uptakes of both substrates. The H(+)-gradient-driven uptake of both substrates was trans-stimulated by L-LHA, D-LHA, L-valine hydroxy analogue or L-lactate, respectively. In the presence of a Na(+)-gradient the uptake of tracer L-lactate was trans-stimulated only after preloading the vesicles with L-lactate but not by L-LHA. It can be concluded that the H(+)-gradient-driven transport of L- or D-hydroxy analogues of branched chain amino acids and L-lactate across the human intestinal brush border membrane is mediated by the same carrier.


SARS-CoV-2 Infection in Children with Idiopathic Nephrotic Syndrome: A Multicentric Study.
A multicenter retrospective study was conducted to assess the clinical spectrum of 30 severe acute respiratory syndrome coronavirus (SARS-CoV-2)-positive children with idiopathic nephrotic syndrome. Difficult to treat nephrotic syndrome was found to be a high-risk group with a high incidence of acute kidney injury and mortality.


The role of the cyclin D1-dependent kinases in ErbB2-mediated breast cancer.
Intact cyclin D1 functions are essential for transformation by erbB2 in tissue culture and murine models. Because cyclin D1 may alter cell proliferation through a variety of mechanisms, we used transgenic models and human tumor samples to particularly address the role of cyclin D1-cyclin-dependent kinases in transformation by erbB2. The p16 tumor suppressor specifically blocks cyclin-dependent kinase 4 and 6 activity. Here we show that an MMTV-p16 transgene blocked tumorigenesis by erbB2, demonstrating that deregulation of the cyclin-dependent kinase partner of cyclin D1 is an essential target of erbB2. ErbB2 overexpression was a determining factor in deregulation of cyclin D1-cdk4/6 interactions because neither transgenic cyclin D1 nor loss of p16 accelerated tumorigenesis in MMTV-erbB2-transgenic mice. ErbB2 was also a deciding factor in deregulation of cyclin D1-cdk4/6 in human tumors because no loss of pRb or p16 was found in tumors overexpressing erbB2, although erbB2-negative invasive breast adenocarcinomas frequently lacked expression of p16 or pRb. We conclude that deregulation of cyclin D1-Cdk4/6 interactions is a critical target of erbB2 function in human and mouse breast tumors, and erbB2's overexpression may be sufficient to deregulate cyclin D1-cdk4/6 activity in breast cancer.


[Cell surface hydrophobicity as an indicator of other virulence factors in Candida albicans].
To evaluate virulence factors involved in the adhesion process, such as cell surface hydrophobicity (CSH), adherence to plastic capacity, adherence capacity to buccal epithelial cells (BEC), and biofilm formation, in 17 strains of C. albicans isolated from bronchial aspirates of critically ill patients. The CSH of the strains of C. albicans was determined using the MATH method, a microbial adhesion to hydrocarbons test. The study of adherence to plastic was performed in microtitre plates in accordance with Christensen's technique. Biofilm formation was studied in polystyrene microtitre plates, according to the method of Ramage. Adherence to BEC was evaluated by quantifying the percentage of adhered yeasts to cells. All the strains studied showed factors directly involved in adhesion, with variability in the degree of expression among them. Medium-high levels of CSH were found in 52.9% of the strains. The percentage of strains with high values in adherence to plastic was 35.3%. The most hydrophobic strains were the most adherent to plastic, with a correlation coefficient of 0.76. Of the 12 biofilm-producing strains, 6 were high producers. These strains had also high levels of CSH and adherence to plastic, with significant results. All the strains studied adhered to BEC, with results ranging widely from 45 to 157 yeasts/100 BEC, with no significant correlation with the rest of the parameters studied, although CSH was seen to be an indispensable prior requisite for adherence to cells. CSH is a variable characteristic in C. albicans and is directly related to adherence to plastic and biofilm formation. Ease in evaluating CSH permits its quantification, and could be used as an indicator of the presence of other determinants of pathogenicity.


Cholinergic stimulation of inositol phosphate formation in bovine adrenal chromaffin cells: distinct nicotinic and muscarinic mechanisms.
The ability of cholinergic agonists to activate phospholipase C in bovine adrenal chromaffin cells was examined by assaying the production of inositol phosphates in cells prelabeled with [3H]inositol. We found that both nicotinic and muscarinic agonists increased the accumulation of [3H]inositol phosphates (mainly inositol monophosphate) and that the effects mediated by the two types of receptors were independent of each other. The production of inositol phosphates by nicotinic stimulation required extracellular Ca2+ and was maximal at 0.2 mM Ca2+. Increasing extracellular Ca2+ from 0.22 to 2.2 mM increased the sensitivity of inositol phosphates formation to stimulation by submaximal concentrations of 1,1-dimethyl-4-phenyl-piperazinium iodide (DMPP) but did not enhance the response to muscarine. Elevated K+ also stimulated Ca2+-dependent [3H]inositol phosphate production, presumably by a non-receptor-mediated mechanism. The Ca2+ channel antagonists D600 and nifedipine inhibited the effects of DMPP and elevated K+ to a greater extent than that of muscarine. Ca2+ (0.3-10 microM) directly stimulated the release of inositol phosphates from digitonin-permeabilized cells that had been prelabeled with [3H]inositol. Thus, cholinergic stimulation of bovine adrenal chromaffin cells results in the activation of phospholipase C by distinct muscarinic and nicotinic mechanisms. Nicotinic receptor stimulation and elevated K+ probably increased the accumulation of inositol phosphates through Ca2+ influx and a rise in cytosolic Ca2+. Because Ba2+ caused catecholamine secretion but did not enhance the formation of inositol phosphates, phospholipase C activation is not required for exocytosis. However, diglyceride and myo-inositol 1,4,5-trisphosphate produced during cholinergic stimulation of chromaffin cells may modulate secretion and other cellular processes by activating protein kinase C and/or releasing Ca2+ from intracellular stores.


A method for estimating the age of coenuri of Taenia multiceps recovered from the brains of sheep.
The age of coenurus metacestodes from the brains of sheep has been estimated by counting the average number of scolices in each cluster on the coenurus wall and relating it to a calculated regression line derived from an examination of cysts of known age. The method has been applied to 17 natural infections and its value is discussed.


Length of hospital stays and financial incentives: evidence from Dutch rehabilitation centers.
Non-linear reimbursement contracts in healthcare have been increasingly used to quantify providers' responses to financial incentives. In the present research, we utilize a large one-off increase in the reimbursement of rehabilitation care to assess to what extent providers are willing to modify their treating behavior to maximize profits. In order to disincentivize the use of short inpatient stays for rehabilitation care, Dutch policy-makers have instated a two-part stepwise tariff-schedule. A lower tariff-schedule is applied for short hospital stays (≤ 14 days), while a higher tariff-schedule is utilized for longer treatments. Switching from one schedule to the other at day 15 of inpatient care leads to a sudden and large increase in tariffs. We show that, for most care-types, patients are seldom treated in an inpatient setting for less than 15 days, while the majority of patients are discharged after the threshold. Therefore, we conclude that the financial incentive at day 15 leads to considerable distortions in treatment. However, instead of discharging all patients at the threshold point where marginal tariffs are maximized, providers tend to continue treatment indicating altruistic behavior. As healthcare payment systems move away from piecewise reimbursement (e.g., fee-for-service arrangements), and services are increasingly 'lumped' together into e.g., DRGs and bundled payments, the likelihood of such discontinuities in tariff-schedules radically increases. Our research illustrates how such discontinuities in reimbursements can lead to distortions in the amount of healthcare provided contributing to the debate on optimal healthcare contracting design.


MR imaging of the vessel wall.
Since the risks for thrombosis are more dependent on plaque composition than on the degree of luminal narrowing, the radiological assessment of atherosclerosis should extend beyond mere depiction of the arterial lumen. High-resolution MRI of the vessel wall can provide important information about the individual makeup of atherosclerotic plaques, potentially enabling early detection and characterization of lesions before narrowing of the vessel lumen occurs. The MR-based assessment of the vessel wall with sufficient spatial resolution and image contrast, however, remains challenging. Requirements include high signal-to-noise ratio, high contrast-to-noise ratio, good signal penetration depth and homogeneous signal throughout the vessel under investigation, as well as imaging protocols encompassing various contrast weightings. Numerous dedicated radiofrequency (RF) coils have been developed to achieve these goals employing either external surface phased-array coils, or alternatively, utilizing intravascular coils to image the vessel wall from inside the vessel and thus being invasive. For the non-invasive approach of imaging with surface coils, the carotid and the right coronary arteries have been favoured since they are of critical importance and since they are relatively superficial structures, accessible from the outside. To detect the early development of plaque and visualize it globally rather than locally, intravascular contrast agents on the basis of ultrasmall particles of iron oxide can be used as a marker of macrophage activity within the plaque. In the long run, it appears likely that the combination of luminal MR angiography with the administration of susceptibility-based contrast agents and subsequent high-resolution MR of detected atherosclerotic lesions with dedicated RF coils could evolve as the diagnostic concept of choice for the assessment of atherosclerotic disease.


Lifetime leisure-time physical activity and the risk of depressive symptoms at the ages of 65-74 years: the FIN-D2D survey.
To investigate the association between lifetime leisure-time physical activity (LTPA) and depressive symptoms (DS) based on the population-based FIN-D2D cross-sectional survey conducted in 2007. Nine hundred twenty seven randomly selected Finnish men and women aged 65-74 years were included in this study. DS were determined with the Beck Depression Inventory (≥10 points), and lifetime historical LTPA frequency from age 15 years onwards was recalled retrospectively. The frequency of LTPA was highest at the ages of 15-24 years in both the DS present and not present groups (14.6 times per month). LTPA frequency was significantly lower among those with DS compared with those without DS at the age of 45-54 (10.0 vs. 11.3, p=0.041), 55-64 (9.6 vs. 12.6, p<0.001), and 65-74 (8.6 vs. 11.9, p<0.001). Also the prevalence of somatic diseases was higher for those with DS compared with participants without DS. This study reveals lifetime LTPA variation and its potential consequences. The relationship found between reduced lifetime LTPA and the occurrence of DS suggest that promoting LTPA throughout one's lifetime would be beneficial in terms of DS and somatic diseases in older age.


Influence of starving on the rat gastric mucosa -- light and electron microscopical findings.
The histology and ultrastructure of the rat gastric mucosa were investigated during 168 hours of starvation. An increased desquamation of individual foveolar cells was found. In the preserved cells of the foveolae, the content of the PAS positive mucosubstances did not change during starvation, and no changes took place in the appearance and in the amount of the mucous granules at the electron microscopic investigation. The number of lipid droplets increased in the mucous foveolar cells within 24 and 48 hours. During starvation the mitochondria (mainly in the parietal cells) were enlarged and contained rare mitochondrial cristae. Some mitochondria were distintegrated and removed by lysosomes. The number of lysosomes (mainly cytosergresomes) was markedly increased n parietal cells. A collapse of the intracellular canaliculi occurred as well as a narrowing in the tubulovesicular profiles. In chief cells the profiles of the granular endoplasmic reticulum and Golgi apparatus were reduced. It was shown that the ultrastructural changes induced by starvation can be interpreted functionally in changed histochemical parameters of the gastric mucosa.


[Analysis of 113 cases with mucinous cystic neoplasms in a single institutional study].
To describe the clinicopathological characteristics and prognostic factors of mucinous cystic neoplasm(MCN). One hundred and thirteen patients with MCN comfirmed by histological examination admitted in People's Liberation Army General Hospital from January 1994 to June 2015 were analyzed, including 21 male and 92 female patients with mean age of (50.2±14.5) years. Forty-eight patients had incidental discovery of their cystic neoplasm for another reason, 65 patients were symptomatic, and abdominal pain or distension was main clinical symptom. Among 113 patients with MCN, 75 cases were MCN with dysplasia, and 38 cases were MCN with invasive carcinoma. Wilcoxon test and χ(2) test, et al were used to analyzed the data, respectively. MCN was mostly located in body and tail of pancreas.Sex, mean age, clinical symptom, tumor size, and CA19-9 showed significant difference between MCN with dysplasia and MCN with invasive carcinoma (all P<0.05). The presence of a solid component (χ(2)=32.460, P=0.000)and main pancreatic duct dilation(χ(2)= 5.729, P=0.022) were significantly associated with malignancy. Fifty-eight patients with dysplasia were followed up, only one patient occurred tumor recurrence. Thirty-one patients with malignancy were followed up, among which there were 22 patients dead, 1-, 3-, 5-year survival of MCN with malignancy was 76.9%, 56.5%, 36.6%, respectively. Lymphatic metastasis and tumor recurrence were important prognostic factors of MCN with malignancy(both P<0.05). MCN is most affected by old female with no specific symptom, most tumors are located at the body and tail of the pancreas.MCN with dysplasia have excellent prognosis underwent surgery. Even with complete resection, the long-term survival of MCN with malignancy is not satisfied.


Heterogeneity among the non-Hodgkin's lymphomas. Implications for autologous bone marrow transplantation with in vitro purging using monoclonal antibodies.
To investigate the possible implications of heterogeneity among the non-Hodgkin's lymphomas for bone marrow purging using complement-fixing monoclonal antibodies to lymphoma-associated antigens, a panel of large cell lymphoma cell lines of diverse phenotypes was treated with monoclonal antibodies DLC-48 and LN-1. An association was demonstrated between the percentage of suppression of colony formation by the cell line and both the percentage of cells staining with the antibody and the intensity of its binding. Flow cytometric analysis of cells surviving treatment with antibody and complement demonstrated that the population that escaped lysis showed weak immunofluorescent staining. Similarly, 40% of the clones derived from cells surviving treatment with antibody and complement stained weakly compared with the parent cell line. For a given fluorescence intensity, cells differed in their susceptibility to treatment. Some cells with moderate to strong staining survived incubation with antibody and complement. In five cases, treatment of bone marrow contaminated with malignant lymphoma cells resulted in complete eradication of even cells with weak staining. In two cases, a population of cells that stained dimly survived treatment with either antibody. DNA-content analysis showed that the cell cycle distribution of cells surviving treatment with DLC-48 or LN-1 and complement was similar to that of cells treated with control antibody 46-1G7 and complement. Phenotypic heterogeneity may hinder efforts to purge malignant lymphoma cells from human bone marrow with complement-fixing monoclonal antibody reagents. Relative resistance to complement-mediated lysis may underlie differences in the susceptibility of cells to treatment and also limit the effectiveness of this technique.


Optimal timing of initiation of oral P2Y12-receptor antagonist therapy in patients with non-ST elevation acute coronary syndromes. Lessons learnt from the ACCOAST-trial.
The optimal time-point of the initiation of P2Y12 antagonist therapy in patients with non-ST elevation acute coronary syndromes (NTSE-ACS) is still a matter of debate. European guidelines recommend P2Y12 as soon as possible after first medical contact. However, the only trial which compared the two strategies did not demonstrate any benefit of pre-treatment with prasugrel before angiography compared to starting therapy after angiography and just prior to percutaneous coronary intervention (PCI). This paper summarizes the results of pharmacodynamic and previous studies, and gives recommendations for the initiation of P2Y12 antagonist therapy in NSTE-ACS in different clinical situations.


Asymptomatic peripheral arterial disease is associated with more adverse lower extremity characteristics than intermittent claudication.
This study assessed functional performance, calf muscle characteristics, peripheral nerve function, and quality of life in asymptomatic persons with peripheral arterial disease (PAD). PAD participants (n=465) had an ankle brachial index <0.90. Non-PAD participants (n=292) had an ankle brachial index of 0.90 to 1.30. PAD participants were categorized into leg symptom groups including intermittent claudication (n=215) and always asymptomatic (participants who never experienced exertional leg pain, even during the 6-minute walk; n=72). Calf muscle was measured with computed tomography. Analyses were adjusted for age, sex, race, ankle brachial index, comorbidities, and other confounders. Compared with participants with intermittent claudication, always asymptomatic PAD participants had smaller calf muscle area (4935 versus 5592 mm(2); P<0.001), higher calf muscle percent fat (16.10% versus 9.45%; P<0.001), poorer 6-minute walk performance (966 versus 1129 ft; P=0.0002), slower usual-paced walking speed (P=0.0019), slower fast-paced walking speed (P<0.001), and a poorer Short-Form 36 Physical Functioning score (P=0.016). Compared with an age-matched, sedentary, non-PAD cohort, always asymptomatic PAD participants had smaller calf muscle area (5061 versus 5895 mm(2); P=0.009), poorer 6-minute walk performance (1126 versus 1452 ft; P<0.001), and poorer Walking Impairment Questionnaire speed scores (40.87 versus 57.78; P=0.001). Persons with PAD who never experience exertional leg symptoms have poorer functional performance, poorer quality of life, and more adverse calf muscle characteristics compared with persons with intermittent claudication and a sedentary, asymptomatic, age-matched group of non-PAD persons.


Cytomegalovirus colitis mimicking ischemic colitis in an immunocompetent host.
Cytomegalovirus (CMV) causes infections in healthy individuals and compromised hosts. In compromised hosts, CMV may cause encephalitis, pneumonia, hepatitis, colitis, and so forth. In immunocompetent hosts, CMV mononucleosis is the most common clinical manifestation and CMV colitis is rare. We present a case of an 82-year-old immunocompetent man who presented with community-acquired bloody diarrhea. A computed tomography scan of the abdomen revealed pan-colitis. His age and abdominal pains suggested ischemic colitis as the cause of his bloody diarrhea. Workup for Clostridium difficile and all enteric pathogens were negative. The patient remained febrile with abdominal pain. During the second week, he underwent sigmoidoscopy for biopsy, which revealed viral inclusions of the Cowdry owl eye inclusion bodies characteristic of CMV. CMV colitis was diagnosed in the patient; he was successfully treated with a course of oral valganciclovir and made an uneventful recovery.


Management of Brugada Syndrome: Thirty-Three-Year Experience Using Electrophysiologically Guided Therapy With Class 1A Antiarrhythmic Drugs.
Information on long-term clinical outcome of patients with Brugada syndrome treated with electrophysiologically guided class 1A antiarrhythmic drugs (AAD) is limited. An aggressive protocol of programmed ventricular stimulation was performed in 96 patients with Brugada syndrome (88% males; mean age, 39.8±15.9 years). Ten patients were cardiac arrest survivors, 27 had presented with syncope, and 59 were asymptomatic. Ventricular fibrillation was induced in 66 patients, including 100%, 74%, and 61% of patients with cardiac arrest, syncope, and no symptoms, respectively. All but 6 of the 66 patients with inducible ventricular fibrillation underwent electrophysiological testing on quinidine (n=54), disopyramide (n=2), or both (n=4). Fifty-four (90%) patients were electrophysiological responders to >1 AAD with similar efficacy rates (≈90%) in all patients groups. Patients with no inducible ventricular fibrillation at baseline were left on no therapy. After a mean follow-up of 113.3±71.5 months, 92 patients were alive, whereas 4 died from noncardiac causes. No arrhythmic event occurred during class 1A AAD therapy in any of electrophysiological drug responders and in patients with no baseline inducible ventricular fibrillation. Arrhythmic events occurred in only 2 cardiac arrest survivors treated with implantable cardioverter-defibrillator alone but did not recur on quinidine. All cases of recurrent syncope (n=12) were attributed to a vasovagal (n=10) or nonarrhythmic mechanism (n=2). Class 1A AAD therapy resulted in 38% incidence of side effects that resolved after drug discontinuation. Our data suggest that electrophysiologically guided class 1A AAD treatment has a place in our therapeutic armamentarium for all types of patients with Brugada syndrome.


Characteristics of hepatocellular carcinoma in Italy.
This study aimed to assess the main features of hepatocellular carcinoma at the time of diagnosis in Italy, particularly in relation to the presence or absence of underlying cirrhosis, hepatitis virus marker patterns, age of the subjects and alpha-foetoprotein values. A total of 1148 patients with hepatocellular carcinoma seen at 14 Italian hospitals in the 1-year period from May 1996 to May 1997 were the subjects of this prevalence study. Both newly diagnosed cases (incident cases) and cases diagnosed before May 1996 but still attending the hospitals during the study period (prevalent cases) were included. We found that 71.1% of cases were positive for hepatitis C virus antibodies but negative for HBsAg; in contrast, 11.5% were negative for anti-HCV but positive for HBsAg; 5.3% were positive for both markers; and 12.1% were negative for both viruses. The mean age of detection was over 60 years, with a younger mean age in HBsAg-positive compared to anti-HCV-positive patients (59.3 years vs. 65.6 years, p<0.01). The male-to-female ratio among HBsAg-positive patients was 10.4:1, in contrast to 2.8:1 among anti-HCV-positive patients (p<0.01). The majority of cases (93.1%) had underlying cirrhosis. Cirrhotic patients were more likely to be anti-HCV positive than non-cirrhotic cases (73.2% vs 43.9%; p<0.01); conversely, absence of hepatitis virus markers was more frequently observed in the non-cirrhotic than in the cirrhotic population (40.9% vs. 10.0%; p<0.01). Overall, the alpha-foetoprotein level was altered (>20 ng/ml) in 57.9% of patients; only 18% of cases presented diagnostic (>400 ng/ml) values. Anti-HCV positivity (O.R. 2.0; CI 95%=1.3-3.1) but not HBsAg positivity (O.R. 1.0; CI 95%=0.6-1.8) was shown to be an independent predictor of the likelihood of altered alpha-foetoprotein values by multivariate analysis. These findings point to differences in the characteristics of the populations infected by hepatitis B and hepatitis C. Factors other than the hepatitis viruses are important in non-cirrhotic patients. A change in the relative prevalence of hepatitis virus markers among hepatocellular carcinoma cases was demonstrated, reflecting a significant change in the rate of HBV endemicity in the Italian population. Finally, the increased trend in the mortality rate from liver cancer in Italy from 4.8 per 100,000 in 1969 to 10.9 in 1994 may reflect the large cohort of subjects infected with HCV via the iatrogenic route during 1950s and 1960s when glass syringes were commonly used for medical treatment.


Relaxation Response and Resiliency Training and Its Effect on Healthcare Resource Utilization.
Poor psychological and physical resilience in response to stress drives a great deal of health care utilization. Mind-body interventions can reduce stress and build resiliency. The rationale for this study is therefore to estimate the effect of mind-body interventions on healthcare utilization. Estimate the effect of mind body training, specifically, the Relaxation Response Resiliency Program (3RP) on healthcare utilization. Retrospective controlled cohort observational study. Major US Academic Health Network. All patients receiving 3RP at the MGH Benson-Henry Institute from 1/12/2006 to 7/1/2014 (n = 4452), controls (n = 13149) followed for a median of 4.2 years (.85-8.4 yrs). Utilization as measured by billable encounters/year (be/yr) stratified by encounter type: clinical, imaging, laboratory and procedural, by class of chief complaint: e.g., Cardiovascular, and by site of care delivery, e.g., Emergency Department. Subgroup analysis by propensity score matched pre-intervention utilization rate. At one year, total utilization for the intervention group decreased by 43% [53.5 to 30.5 be/yr] (p <0.0001). Clinical encounters decreased by 41.9% [40 to 23.2 be/yr], imaging by 50.3% [11.5 to 5.7 be/yr], lab encounters by 43.5% [9.8 to 5.6], and procedures by 21.4% [2.2 to 1.7 be/yr], all p < 0.01. The intervention group's Emergency department (ED) visits decreased from 3.6 to 1.7/year (p<0.0001) and Hospital and Urgent care visits converged with the controls. Subgroup analysis (identically matched initial utilization rates-Intervention group: high utilizing controls) showed the intervention group significantly reduced utilization relative to the control group by: 18.3% across all functional categories, 24.7% across all site categories and 25.3% across all clinical categories. Mind body interventions such as 3RP have the potential to substantially reduce healthcare utilization at relatively low cost and thus can serve as key components in any population health and health care delivery system.


Distribution of fibres reacting with an alpha-endorphin antiserum in the neurohypophysis of Carassius auratus and Cyprinus carpio.
A strong positive immunoreaction with an alpha-endorphin antiserum occurs in two distinct sites of the goldfish and carp neurohypophysis. Fluorescent nerve terminals are found in the laminar nerve processes located in the rostral pars distalis, but the immunocytological reaction is mainly localised on the nerve processes of the posterior neurohypophysis lying between the intermediate lobe cells. Almost all the digitations of the neurohypophysis are strongly fluorescent. The immunoreactive fibres probably originate from the hypothalamus, where perikarya displaying the same immunoreaction have been found in the pars lateralis of the nucleus lateralis tuberis and in some minor centres. The possibility that the immunoreactive substances revealed on the neurohypophyseal processes may originate in the intermediate lobe cells is also discussed. It has now to be established if this hypothalamo-hypophyseal system contains a substance with endorphic properties or only some immunologically related substance devoid of the corresponding physiological activities.


Identification of thioredoxin target protein networks in cardiac tissues of a transgenic mouse.
The advent of sensitive and robust quantitative proteomics techniques has been emerging as a vital tool for deciphering complex biological puzzles that would have been challenging to conventional molecular biology methods. The method here describes the use of two isotope labeling techniques-isobaric tags for relative and absolute quantification (iTRAQ) and redox isotope-coded affinity tags (ICAT)-to elucidate the cardiovascular redox-proteome changes and thioredoxin 1 (Trx1)-regulated protein network in cardiac-specific Trx1 transgenic mouse models. The strategy involves the use of an amine-labeling iTRAQ technique, gauging the global proteome changes in Trx1 transgenic mice at the protein level, while ICAT, labeling redox-sensitive cysteines, reveals the redox status of cysteine residues. Collectively, these two quantitative proteomics techniques can not only quantify global changes of the cardiovascular proteome but also pinpoint specific redox-sensitive cysteine sites that are subjected to Trx1-catalyzed reduction.


A highly sensitive flow-through amperometric immunosensor based on the Peroxidase chip and enzyme-channeling principle.
A concept based on the Peroxidase-chip (P-chip), antibody co-immobilization, competitive and enzyme-channeling principle was exploited to develop an integrated flow-through amperometric biosensor for detection of environmental pollutants such as s-triazine herbicides. In this concept, recombinant peroxidase is immobilized on the gold electrode (P-chip) in such a way that direct electron transfer is achieved. The recognition and quantitation the target analyte is realized through the competition between the simazine-glucose oxidase (GOD) conjugate and free simazine for the binding sites of the monoclonal antibody co-immobilized with peroxidase on the gold electrode. The arrangement allows to generate a specific signal in the presence of glucose through the channeling of H2O2 produced by GOD conjugate bound to the antibody. The immunosensor exhibited 50% signal decrease (IC50 value) at approximately 0.02 microg l(-1). A concentration of 0.1 ng l(-1) gave a signal clearly distinguishable from the blank whereas the ELISA using the same antibody had a typical detection limit of about 1 microg l(-1), which is four orders of magnitude higher compared to the presented biosensor system. The results demonstrated that gene engineering biomolecules, in this case recombinant peroxidase, might be attractive reagents for the development of electrochemical immunosensors.


Deriving a high-quality daily dataset of large-pan evaporation over China using a hybrid model.
Global warming is expected to increase the atmospheric evaporative demand and make more surface water for evapotranspiration, aggerating water sources' social and ecological shortage. Pan evaporation, as a routine observation worldwide, is an excellent metric to indicate the response of terrestrial evaporation to global warming. However, several non-climatic effects, such as instrument upgrades, have destroyed the homogenization of pan evaporation and limited its applications. In China, 2400s meteorological stations have observed daily pan evaporation since 1951. The observed records became discontinuous and inconsistent due to the instrument upgrade from micro-pan D20 to large-pan E601. Here, combining the Penpan model (PM) and random forest model (RFM), we developed a hybrid model to assimilate different types of pan evaporation into a consistent dataset. Based on the cross-validation test, on a daily scale, the hybrid model has a lower bias (RMSE=0.41 mm day-1) and better stability (NSE=0.94) than the two sub-models and the conversion coefficient method. Finally, we produced a homogenized daily dataset of E601 across China from 1961 to 2018. Based on this dataset, we analyzed the long-term trend of pan evaporation. Pan evaporation showed a -1.23±0.57 mm a-2 downward trend from 1961-1993, primarily caused by decreased pan evaporation in warm seasons over North China. After 1993, the pan evaporation in South China increased significantly, resulting in a 1.83±0.87 mm a-2 upward trend across China. With better homogeneity and higher temporal resolution, the new dataset is expected to promote drought monitoring, hydrological modeling, and water resources management. Free access to the dataset can be found at https://figshare.com/s/0cdbd6b1dbf1e22d757e.


Tumor necrosis factor-α-mediated threonine 435 phosphorylation of p65 nuclear factor-κB subunit in endothelial cells induces vasogenic edema and neutrophil infiltration in the rat piriform cortex following status epilepticus.
Status epilepticus (SE) induces severe vasogenic edema in the piriform cortex (PC) accompanied by neuronal and astroglial damages. To elucidate the mechanism of SE-induced vasogenic edema, we investigated the roles of tumor necrosis factor (TNF)-α in blood-brain barrier (BBB) disruption during vasogenic edema and its related events in rat epilepsy models provoked by pilocarpine-induced SE. SE was induced by pilocarpine in rats that were intracerebroventricularly infused with saline-, and soluble TNF p55 receptor (sTNFp55R) prior to SE induction. Thereafter, we performed Fluoro-Jade B staining and immunohistochemical studies for TNF-α and NF-κB subunits. Following SE, most activated microglia showed strong TNF-α immunoreactivity. In addition, TNF p75 receptor expression was detected in endothelial cells as well as astrocytes. In addition, only p65-Thr435 phosphorylation was increased in endothelial cells accompanied by SMI-71 expression (an endothelial barrier antigen). Neutralization of TNF-α by soluble TNF p55 receptor (sTNFp55R) infusion attenuated SE-induced vasogenic edema and neuronal damages via inhibition of p65-Thr435 phosphorylation in endothelial cells. Furthermore, sTNFp55R infusion reduced SE-induced neutrophil infiltration in the PC. These findings suggest that impairments of endothelial cell functions via TNF-α-mediated p65-Thr 485 NF-κB phosphorylation may be involved in SE-induced vasogenic edema. Subsequently, vasogenic edema results in extensive neutrophil infiltration and neuronal-astroglial loss.


Active components of Bupleurum chinense and Angelica biserrata showed analgesic effects in formalin induced pain by acting on Nav1.7.
Pain is an unpleasant sensory and emotional experience, often accompanied by the occurrence of a variety of diseases. More than 800 kinds of traditional Chinese medicines (TCM) has now been reported for pain relief and several monomers have been developed into novel analgesic drugs. Bupleurum chinense and Angelica biserrata were representatives of the TCM that are currently available for the treatment of pain. The study aims to detect the potential analgesic activity of each monomer of Bupleurum chinense and Angelica biserrata and to explore whether Nav1.7 is one of the targets for its analgesic activity. In this study, five monomers from Bupleurum chinense (Saikosaponin A, Saikosaponin B1, Saikosaponin B2, Saikosaponin C, Saikosaponin D) and five monomers from the Angelica biserrata (Osthole, Xanthotoxin, Imperatorin, Isoimperatorin, Psoralen) were examined by whole-cell patch-clamp on Nav1.7, which was closely associated with pain. Classical mouse pain models were also used to further verify the analgesic activity in vivo. The results showed that monomers of Saikosaponins and Angelica biserrata all inhibited the peak currents of Nav1.7, indicating that Nav1.7 might be involved in the analgesic mechanism of Saikosaponins and Angelica biserrata. Among them, Saikosaponin A and Imperatorin showed the strongest inhibitory effect on Nav1.7. Furthermore, both Saikosaponin A and Imperatorin showed inhibitory effects on thermal pain and formalin-induced pain in phase II in vivo. The results provide valuable information for future studies on the potential of TCM in alleviating pain.


Psychosocial experiences of parents of infants with hypoplastic left heart syndrome in the PICU.
To evaluate the psychosocial status of mothers and fathers of infants with hypoplastic left heart syndrome while in the PICU. A retrospective study combining interviews and psychometric testing of parents. Tertiary hospital PICU. Twenty-nine parents (16 mothers and 13 fathers) of surviving children. A semistructured face-to-face interview was conducted to explore parental experiences, and a Structured Clinical Interview for Diagnosis-Clinical Version (posttraumatic stress disorder module) was conducted to determine the possibility of an acute stress disorder or a posttraumatic stress disorder. All parents reported multiple stresses which commenced with their infant's diagnosis and endured throughout their infant's time in PICU. The Structured Clinical Interview for Diagnosis revealed that acute stress disorder or posttraumatic stress disorder developed in 24 parents (83%). Of 18 parents whose infants were diagnosed with hypoplastic left heart syndrome in utero, eight of nine mothers (88%) and six of nine fathers (66%) had posttraumatic stress disorder. Of 11 parents whose infants were diagnosed with hypoplastic left heart syndrome postbirth, six of seven mothers had acute stress disorder and one mother had posttraumatic stress disorder, and of four fathers, two fathers had acute stress disorder and one father had posttraumatic stress disorder. The prevalence of parental stress-related disorder was not different between mothers and fathers (p = 0.85). Only five parents were free of traumatic stress-related illness. Parents also experienced losses. Many parents were marginalized from their infant's care by the environment of PICU. Fifty percent of mothers experienced difficulties with parental-infant bonding. Ten parents (34%) began the process of adaptation to their infant's hypoplastic left heart syndrome and were assisted by the support and sensitivity of staff or had discovered other resources. All parents of surviving infants with hypoplastic left heart syndrome in PICU, irrespective of timing of diagnosis, experienced numerous stresses and losses, and the majority exhibited clinical levels of traumatic stress. Receiving the diagnosis itself is very traumatic and is compounded by the environment of the PICU which alienates parents from their infants and interferes with parent-infant bonding. Parental adaptation to this situation can be assisted by staff.


Determination by modified RT-PCR of transcript amounts from genes involved in sex-steroid synthesis in chicken organs including brain.
Sex steroid hormones in mammals have shown to be synthesized not only in gonads but also in non-steroidogenic organs such as the brain. Steroid hormones in the brain were indicated to be involved in sex behavior and brain differentiation. In avian species, an experimental injection of androgen into the brain suggested the existence of a steroidogenic pathway. However, no studies have demonstrated the expression of genes involved in such a steroidogenic pathway in the brain, or in other non-steroidogenic organs of birds such as the liver and kidney. In this study, we have modified the RT-PCR procedure to analyze the expression of the steroidogenic genes, P-450scc, 3beta-HSD, P-450c17, 17beta-HSD and P-450arom in non-steroidogenic organs of chicken including the brain. The RT-PCR has demonstrated the presence of mRNAs from genes in non-steroidogenic as well as steroidogenic organs of chicken. The amounts of mRNAs from these genes (except for P-450c17) among the non-steroidogenic organs were found to be greatest in the brain.


Growth regulatory peptide production by human breast carcinoma cells.
The mechanisms by which human breast cancers regulate their own growth have been studied by us in an in vitro model system. We showed that specific growth factors (IGF-I, TGF alpha, PDGF) are secreted by human breast cancer cells. A variety of experiments suggest that they are involved in tumor growth and progression. These activities are induced by estradiol in hormone-dependent breast cancer cells and secreted constitutively by estrogen-independent cells. Concentrates of conditioned medium derived from breast cancer cells can induce the growth of hormone-dependent cells in vivo in athymic nude mice. Hormone-dependent breast cancer cells also secrete TGF beta. TGF beta is growth inhibitory. Growth inhibitors such as antiestrogens or glucocorticoids increase TGF beta secretion. An antiestrogen-resistant mutant of MCF-7 cells does not secrete TGF beta when treated with antiestrogen, but is growth inhibited when treated with exogenous TGF beta. Thus, TGF beta functions as a negative autocrine growth regulator and is probably responsible for some of the growth inhibitory effects of antiestrogens.


Insights into early postoperative acute kidney injury following lung transplantation.
Acute kidney injury (AKI) is a common complication after lung transplantation (LT) and is associated with higher cost and mortality. We sought to evaluate the incidence of postoperative AKI, defined as AKI within 14 days of transplant, and identify associated perioperative factors. We conducted a single-center, retrospective review of 153 lung transplant recipients. Postoperative AKI was determined using the RIFLE (Risk, Injury, Failure, Loss, End Stage) criteria. Perioperative covariates and their association with postoperative AKI were analyzed using Cox proportional hazards. Kaplan-Meier survival curves were constructed to evaluate patient survival at 1 year and data finalization. A sub-analysis was performed evaluating factors associated with early AKI (within 48 h of transplant) and late AKI. Postoperative AKI occurred in 36.6% of patients with 51.8% of cases occurring within 48 h of LT. Recipient race, transplant type, cardiopulmonary support, and red blood cell administration were associated with postoperative AKI. Survival was significantly lower in patients with postoperative AKI following LT. Postoperative AKI within 2 weeks of lung transplant is associated with lower short- and long-term survival. Perioperative factors associated with postoperative AKI may be potential points of intervention to minimize AKI development in the future.


Natural iron-aluminosilicate as potential solid precursor for supplementary cementitious materials: A comparative study with other aluminosilicates.
The objective of this study was to investigate the impact of the geographic and climatic conditions on laterites properties and on geopolymerization based-laterite. Four different laterite deposits in the four geographical zones of Cameroon were studied. This included the center, north, south and west corners of Cameroon, having chemical composition of SiO2 + Al2O3 + Fe2O3 = 88.94, 87.6, 89.13 and 78.97%, respectively. The center and south laterites from the black forest, with high pluviometry and relative humidity, show significant amounts of Fe2O3. While the west laterite from grass field - mountainous areas and the north-laterite from plain arid and semi-arid climate still show lower iron concentrations. The IR absorption bands of the different laterites appear between 1007 and 1047 cm-1; characteristic bands of aluminosilicate. The BET (Brunauer-Emmett-Teller) Specific surface area values are comprised in the range of [21.9, 24.1 m2/g] for non-calcined laterite and between [45.6 and 123.5 m2/g] for laterites calcined at 550 °C and 575 °C. The main particle size values are 5.71, 6.37, 7.43 and 8.45 μm for center-laterite, west-laterite, north laterite and south-laterite, respectively. Although, they differ in the degree of laterization, all the laterites present almost total conversion to geopolymers, due to the presence of amorphous kaolinite and reactive goethite. However, the iron content has significant impact on the globular microstructure. The particle size of laterites, their high values of BET surface area and their significant reactivity make them promising substitutes to metakaolin and other supplementary cementitious materials.


Assessment of reasons for referral and activities of hospital palliative care teams using a standard format: a multicenter 1000 case description.
The many benefits of hospital palliative care teams (PCTs) are well known. However, their specific activities have not been fully clarified, and no standardized methods for reporting PCT activities are available. The aim of this study was to investigate, through the use of a standard format, the activities performed by hospital PCTs in Japan. This was a prospective observational study. A total of 21 hospital PCTs were included in this study, and each recruited approximately 50 consecutively referred patients. Participating PCTs filled in a standard form for reporting activities. We obtained data from 1055 patients who were referred to PCTs. Of the 1055 patients, 1005 patients (95%) had cancer. The median number of reasons for referral and problems identified by PCTs was two (0-22) and four (0-18), respectively. The two major reasons for referral were pain (63%) and anxiety/depression/grief/emotional burden (22%). The major recommendations were pharmacological treatment (74%), care for the patient's physical symptoms (49%), and support for patient's decision making (38%). The major activities performed by the PCTs were comprehensive assessment (90%), care for the patient's physical symptoms (77%), and pharmacological treatment (74%). The components of hospital PCT activities were successfully measured using the Standard Format for Reporting Hospital PCT Activity. The results of this study and the format for reporting hospital PCT activity could be effective in improving hospital PCT practice and for the education of new hospital PCT members.


Value of the MELD score for the assessment of pre- and post-liver transplantation survival.
The MELD score has now been implemented in the United States for liver allocation, but it has not been validated in Europe. Its association with posttransplant outcome is unclear. Optimal cutoff values of MELD and Child-Pugh scores to predict death on the liver waiting list were defined in a series of 137 cirrhotic patients listed for liver transplantation. Six-month actuarial survival while on the waiting list was 90% with a Child-Pugh <11 and MELD <17, whereas it decreased progressively to 40% at 6 months after listing for those having a Child-Pugh and MELD score >10 and >16. Analysis of a series of 112 patients (85 chronic liver disease and 27 hepatocellular carcinoma) revealed no change in MELD value at the time of transplantation compared to the score at the time of listing (mean +/- SD: 15.5 +/- 7.7 vs 15 +/- 5.8) with a mean waiting time of 118 days. Using either the optimal cutoff for MELD score (<17 or >16) or seven different strata (3 to 7, 8 to 10, 11 to 13, 14 to 16, 17 to 19, 20 to 22, 23 to 39), whether measured at listing or just before liver transplantation, there was no significant difference (chi(2) 4.97, P = .58) in survival: 82.7% and 63% at 6 and 60 months, overall. Our data confirm that the MELD score with only three parameters is as good as the Child-Pugh score to predict mortality on the Eurotransplant waiting list. The optimal cutoff to assess higher priority for the bad category is >16. There was no negative impact on short- or long-term prognosis of the bad categories of MELD.


Chromium (VI) detection by microbial carbon dots: Microwave synthesis and mechanistic study.
Functionalized carbon dots (CDs) derived from Citrobacter freundii bacterial cells were used for selective detection of Cr(VI). A microwave-heating-based green synthesis approach is adopted to produce functionalized CDs from C. freundii bacterial cells (CF-CDs). The reaction was carried out in a 500 W microwave digester at 200°C for 20 min. The supernatant was filtered with a 0.2 µm filter and highly monodisperse CDs were obtained. Inherent functionalization of CF-CDs with nitrogen and oxygen-containing functional groups made them extremely selective toward Cr(VI) with a lower limit of detection of ~1.7 ppm. More importantly, CF-CDs could distinguish between Cr(VI) and Cr(III), which is highly desirable for practical applications. The fabricated sensor had a dual linear response range between 0 and 50 µM and 50-250 µM. The synthesized CDs were inherently functionalized which made them highly selective for Cr(VI) detection. CF-CDs also possess high stability over long storage period. This study reports the facile synthesis and characterization of a highly selective sensor for Cr(VI). As opposed to similar CDs sensors reported for Cr(VI), based on inner-filter effect, CF-CDs exhibit a strong, specific interaction toward Cr(VI), indicated by the large binding constant.


Serum albumin, serum antiproteases and polymorphonuclear leucocyte neutral collagenolytic protease in the tear fluid of normal healthy persons.
Using a micro-electroimmune assay and a 'non-reflex stimulating' method to collect normal tear fluid, the concentration of the serum proteins albumin (S-A), alfa-1-antitrypsin (alpha 1A), alfa-1-antichymotrypsin (alpha 1X) and alfa-2-macroglobulin (alpha 1M) was determined in the tear fluid of 53 normal persons. In addition, the tear fluid was tested for the content of polymorphonuclear leucocyte neutral collagenolytic protease (PML-c-ase). PML-c-ase and alpha 1X could not be detected in any tear samples. The concentrations of S-A, alpha 1A and alpha 1M were constant over time, and showed no variation with sex, age or between the right and left eyes. There was no correlation between the concentration levels of the detected proteins in the tear fluid and correlation between tear fluid levels and corresponding serum concentration levels of the proteins could not be shown.


Prevalence and mechanisms of fosfomycin resistance among KPC-producing Klebsiella pneumoniae clinical isolates in China.
The threat of antibiotic resistance has increased dramatically in recent years. Fosfomycin, an old antibiotic agent, has been re-introduced to fight infections caused by Klebsiella pneumoniae carbapenemase-producing K. pneumoniae (KPC-KP). However, the trend of fosfomycin resistance among KPC-KP strains is increasing. In this study, 80 KPC-KP clinical isolates were collected from three teaching hospitals during 2014-2017 in China and were subjected to whole-genome sequencing (WGS). The fosfomycin resistance phenotype and resistance mechanisms were investigated by antimicrobial susceptibility testing and carbon source growth test, respectively. Among all KPC-KP strains, 80.0% (64/80) were resistant to fosfomycin and 36.3% (29/80) were positive for the mobile fosfomycin resistance gene fosA3. Among the 63 strains that were unable to grow in M9 basic medium with glycerol-3-phosphate (G3P) as the sole carbon source (mediated by mutation of the target gene glpT), there was no significant difference regarding the MIC distribution of fosfomycin between fosA3-positive and fosA3-negative strains (P = 0.577). Among the 50 strains that were negative for fosA3 but positive for fosA, the fosfomycin MICs of strains unable to grow in M9 basic medium with G3P as the sole carbon source were significantly higher (P < 0.001) than in strains that were able to grow in M9 basic medium with G3P as the sole carbon source. Our findings indicate that fosfomycin resistance among KPC-KP in China is an emerging problem and the two major mechanisms of resistance identified were plasmid-mediated fosfomycin resistance gene fosA3 and mutation of the target gene glpT.


All-optical monitoring of excitation-secretion coupling demonstrates that SV2A functions downstream of evoked Ca2+ entry.
SV2A, an essential transporter-like synaptic vesicle protein, is a major target for antiepileptic drugs and a receptor for clostridial neurotoxins including Botox. While SV2A is required for normal levels of evoked neurotransmitter release, the mechanism underlying this role remains unclear. Here, we introduce a new chemogenetic approach for all-optical monitoring of excitation-secretion coupling, and we demonstrate its use in characterizing the SV2A knockout (KO) phenotype in cultured hippocampal neurons. This method employs the HaloTag system to target a robust small-molecule Ca2+ indicator, JF646 -BAPTA, to the presynaptic compartment. The far-red fluorescence of this indicator enables multiplexing with the fluorescent glutamate sensor iGluSnFR for detection of presynaptic Ca2+ influx and glutamate release at the same axonal boutons. Evoked glutamate release probability was reduced in SV2A KO neurons without a change in presynaptic Ca2+ entry, suggesting that SV2A supports vesicle fusion by increasing the functional availability, or efficiency, of the Ca2+ -regulated membrane fusion machinery. KEY POINTS: One of the most prescribed antiepileptic medications, levetiracetam, acts by binding a protein of uncertain molecular function. This transporter-like protein, SV2A, is trafficked to synaptic vesicles and acts to support neurotransmitter release, but the mechanism underlying this function has not been determined In this study, we sought to establish whether SV2A changes Ca2+ signalling at nerve terminals, which is a key regulatory system for synaptic vesicle exocytosis. To do so, we adapted new chemogenetic tools to perform all-optical measurements of presynaptic Ca2+ and glutamate release in neurons lacking SV2A. Our measurements showed that loss of SV2A reduces glutamate release without reducing Ca2+ influx at hippocampal nerve terminals, demonstrating that SV2A increases the likelihood that Ca2+ will trigger synaptic vesicle fusion.


Bladder epithelial hyperplasia in F344 rats after intravesical instillation of the antitumor chemotherapeutic agents Adriamycin and mitomycin C.
The effects of intravesical instillation of the antineoplastic antibiotics, Adriamycin or mitomycin C, on the urinary bladder epithelium of female F344 rats were evaluated using a combined immunohistochemical and morphological approach. Four weeks treatment with Adriamycin or mitomycin C induced an increase of DNA synthesis and was associated with simple hyperplasia characterized by elevated nuclear cytoplasmic ratios, cytomegaly and pleomorphism. Under the scanning electron microscope (SEM), luminal cell surface alterations such as pleomorphic microvilli were observed. Severity of the lesions was greatest in the Adriamycin group and although treatment of the saline vehicle alone also brought about development of simple hyperplasia, this was very slight and not accompanied by cellular pleomorphism. The present results demonstrated that intravesical instillation of antineoplastic agents can cause a proliferative response and cytotoxicity after only short-term treatment and suggest that this chemotherapy could itself play a possible role in promotion of bladder carcinogenesis or cancer development.


Explant culture of human colon.
Human colonic epithelium has been cultured as explants in a chemically defined medium for periods of 1 to 20 days. The viability of the explants was shown by the preservation of the ultrastructural features of the colonic epithelial cells and by active incorporation of radioactive precursors into cellular DNA and protein. A progressive decrease in the number of goblet cells, decrease in the depth of the crypts, and a change from a columnar to a cuboidal epithelium were observed. After 20 days in culture the colonic mucosa consisted of a single layer of cuboidal epithelial cells and a few glands. The ability to maintain colonic mucosa in culture was subject to both intra- and interindividual variation. Cultured human colonic mucosa also activated a chemical procarcinogen, benzo[a]pyrene, into metabolites which bound to cellular DNA. A 100-fold interindividual variation in this binding was observed.


The binding of Mn2+ to bovine plasma protein C, des(1-41)-light chain protein C, and activated des(1-41)-light chain activated protein C.
The binding isotherms of Mn2+ to bovine plasma protein C (PC), des(1-41)-light chain protein C (GDPC), and activated GDPC (GDAPC) have been measured. PC contains 14-16 total Mn2+ binding sites, a value that is reduced to approximately 7-8 in the presence of NaCl. The average Kd of the latter sites is 230 +/- 30 microM. Upon removal of a 41-residue peptide from the amino terminus of the light chain of PC, and, concomitantly, all of the gamma-carboxyglutamic acid residues, the resulting protein, GDPC, possesses a single Mn2+ site of Kd = 120 +/- 20 microM. Activation of GDPC to GDAPC results in a slight lowering of the Kd for the single Mn2+ binding site to 53 +/- 8 microM, a value that is essentially unchanged in the presence of monovalent cations, a competitive inhibitor of the enzyme, or an active site directed affinity label. The Mn2+ on GDAPC is displaced by Ca2+, suggesting that the protein binding site for these two divalent cations is the same. These studies establish that Mn2+ is a suitable spectroscopic probe for the Ca2+ binding site of GDAPC, and that the divalent cation site is separate from the monovalent cation site(s) and the active site of the enzyme.


Artemisinin-based combination therapy availability and use in the private sector of five AMFm phase 1 countries.
In 2009, the Global Fund to Fight AIDS, Tuberculosis and Malaria established the Affordable Medicines Facility-malaria (AMFm) in order to increase access to quality-assured artemisinin combination therapy (QAACT). AMFm Phase 1, which includes nine pilot programmes in eight countries, was launched in 2009. The objective of this study was to assess anti-malarial stock and purchase patterns at private outlets in five AMFm Phase 1 countries in regard to three of the core AMFm goals: increase the affordability of QAACT, increase the availability of QAACT, and crowd out artemisinin monotherapies and other substandard therapies. The study was conducted between April and May 2012 and included interviews with personnel in 598 private pharmaceutical outlets in Ghana, Kenya, Nigeria, Tanzania, and Uganda. Questionnaires were administered at private retail outlets and the data were analyzed to assess within- and between-country differences in QAACT price, availability, and popularity. AMFm medications were less expensive than their non-AMFm counterparts, yet prices for both types were above country-specific suggested retail prices. Market penetration of AMFm QAACT in both urban and rural areas was high, although stock-outs of both AMFm and non-AMFm products were more common in rural compared with urban outlets in Ghana and Kenya (p = 0.0013). Government recommendation was the most significant factor influencing anti-malarial stock choices in urban (41.5%) and rural (31.9%) outlets. The three top-selling anti-malarials reported for both urban and rural areas in each country were, with the exception of rural Uganda and urban Nigeria, combination therapies. Results from this study indicate that the AMFm has not fully achieved its affordability and crowd-out objectives. Still, the final purchase price of AMFm QAACT was substantially lower than non-AMFm equivalents. Moreover, for both urban and rural areas, AMFm QAACT availability was found to be high, and the various forms of QAACT were the best-selling products among all anti-malarials. These findings suggest a continued need for initiatives like the AMFm that improve the affordability and accessibility of QAACT. Similar programmes may be especially effective if employed in combination with rapid diagnostic testing to ensure the appropriate use of these products.


Presenting ADAMTS13 on a TTP-associated MHC.
In this issue of Blood, Sorvillo et al investigate possible molecular triggers leading to idiopathic, autoimmune thrombotic thrombocytopenic purpura (TTP) by identifying naturally processed A Disintegrin And Metalloprotease with ThromboSpondin type 1 motif 13 (ADAMTS13)-derived peptides presented on human dendritic cells.


Sympathetic baroreflex sensitivity is inversely related to vascular transduction in men but not women.
Sympathetic baroreflex sensitivity (BRS) is a measure of how effectively the baroreflex buffers beat-to-beat changes in blood pressure through the modulation of muscle sympathetic nerve activity (MSNA). However, current methods of assessment do not take into account the transduction of sympathetic nerve activity at the level of the vasculature, which is known to vary between individuals. In this study we tested the hypothesis that there is an inverse relationship between sympathetic BRS and vascular transduction. In 38 (18 men) healthy adults, continuous measurements of blood pressure, MSNA and superficial femoral artery diameter and blood flow (Doppler ultrasound) were recorded during 10 min of rest. Spontaneous sympathetic BRS was quantified as the relationship between diastolic pressure and MSNA burst incidence. Vascular transduction was quantified by plotting the changes in leg vascular conductance for 10 cardiac cycles following each burst of MSNA, and taking the nadir. In men, sympathetic BRS was inversely related to vascular transduction (r = -0.49; P = 0.04). However, this relationship was not present in women (r = -0.17; P = 0.47). To conclude, an interaction exists between sympathetic BRS and vascular transduction in healthy men, such that men with high sympathetic BRS have low vascular transduction and vice versa. This may be to ensure that blood pressure is regulated effectively, although further research is needed to explore what mechanisms are involved and examine why this relationship was not apparent in women.NEW & NOTEWORTHY Evidence suggests that compensatory interactions exist between factors involved in cardiovascular control. This study was the first to demonstrate an inverse relationship between sympathetic BRS and beat-to-beat vascular transduction. Those with low sympathetic BRS had high vascular transduction and vice versa. However, this interaction was present in young men but not women.


Characteristics of glomerular lesions in hepatitis B virus infection.
In an attempt to clarify a participation of hepatitis B virus (HBV) in the development of hepatic glomerulopathy in adults, kidney specimens obtained from 151 patients with liver diseases were studied. Although mesangial proliferation was more severe in patients with chronic hepatitis or liver cirrhosis than in those with acute hepatitis, no significant difference was observed between 82 serum hepatitis B surface antigen (HBsAg)-positive (HBV-related group) and 69 negative patients (HBV-nonrelated group). However, double contours of the glomerular capillary walls were observed more often in the former group (18/82, P less than .01), especially in the HBeAg-positive subgroup (8/24, P less than .001), than in the latter (3/69). In addition, glomerular capillary spike formation or a bubblelike appearance was observed in seven patients of the former group. Of these, all five patients examined were HBeAg-positive in their serum. By electron microscopic studies, subendothelial dense deposits and mesangial interpositions were observed more frequently in the HBV-related group, and subepithelial deposits were found only in the HBeAg-positive subgroup. The immunofluorescence study revealed IgA-dominant mesangial deposition in both HBV-related and nonrelated groups. As for the capillary wall deposits, however, IgG was dominant in 13 of the HBV-related group, but only one of the nonrelated group (P less than .01). Furthermore, one patient in the HBV-related group showed capillary wall-dominant HBeAg combined with IgG deposition.(ABSTRACT TRUNCATED AT 250 WORDS)


Recurrent Diffuse Pigmented Villonodular Synovitis in the Hand of a Pediatric Patient: A Case Report.
Diffuse pigmented villonodular synovitis is characterized by synovial inflammation and hemosiderin deposition. It mainly occurs in adults, with the hip and knees being the most common sites of involvement. It is associated with high recurrence rates, with open synovectomy being the most common treatment method to avoid recurrences. Few cases of diffuse pigmented villonodular synovitis have been reported in pediatric patients, especially in uncommon locations such as the hand. This case presents pathology-confirmed diffuse pigmented villonodular synovitis in the hand of a pediatric patient with multiple recurrences despite adequate surgical margins. The patient underwent mass excision with adjuvant radiation treatment after his last recurrence, with excellent functional outcomes and no recurrence at the five-year follow-up.


Epidemiology of childhood-onset type 1 diabetes in Azerbaijan: Incidence, clinical features, biochemistry, and HLA-DRB1 status.
Determine the incidence and typology of diabetes in children in Azerbaijan. Clinical features, C-peptide, autoantibodies (glutamic acid decarboxylase 65 (GAD65) and islet antigen 2 (IA-2)), and HLA-DRB1 status were studied in 106 subjects <18 years of age who were recently diagnosed. 104 cases were consecutive. Incidence was determined for Baku and Absheron regions, where ascertainment is estimated to be essentially 100%. 104 of the 106 (98%) were diagnosed with type 1 diabetes, one with type 2 diabetes and one with atypical diabetes. Type 1 diabetes incidence in Baku City and Absheron was 7.05 per 100,000 population <15 years per year. Peak age of onset was 10 years. There was a slight male preponderance (male:female 1.17:1), and no temporal association with seasons. Almost all type 1 diabetes subjects presented with classic symptoms including a high incidence (58%) of diabetic ketoacidosis. 86% presented with low C-peptide values (<0.13 nmol/L, <0.40 ng/mL) and 74% were positive for at least one type 1 diabetes-related autoantibody. Azerbaijan has a moderate type 1 diabetes incidence and clinical, biochemical and genetic features similar to that in European populations.


Prevalence of Aminoglycoside Resistance Genes and Molecular Characterization of a Novel Gene, aac(3)-IIg, among Clinical Isolates of the Enterobacter cloacae Complex from a Chinese Teaching Hospital.
Members of the Enterobacter cloacae complex are important opportunistic human pathogens capable of causing a wide variety of infections. During recent decades, aminoglycoside-resistant E. cloacae complex isolates have increasingly been reported and have become a major concern. Here, we employed high-throughput sequencing in combination with specific PCR assays to investigate the prevalence of aminoglycoside resistance genes among 170 isolates of the E. cloacae complex collected from a teaching hospital in Wenzhou, China. A total of 12 known genes [aphA-1, strA, strB, aac(6')-IIc, aadA2, aac(3)-IId, aadB, aadA1, rmtB, armA, aadA5, and aac(6')-Ie-aph(2'')-Ia] and 1 novel gene [aac(3)-IIg] were identified, with aphA-1 (71.18%), strA (55.29%), and strB (52.35%) being the most prevalent, and aac(3)-IIg was detected with a positive rate of 21.76% (37/170). The aac(3)-IIg gene was 810 bp in length and encoded a protein that shared 72 to 78% identities with previously known AAC(3)-II aminoglycoside 3-N-acetyltransferases. The MICs of gentamicin and tobramycin were 512 μg/ml and 64 μg/ml, respectively, when aac(3)-IIg was cloned into Escherichia coli DH5α. All aac(3)-IIg-positive isolates exerted broad aminoglycoside resistance profiles, mediated by the coexistence of multiple resistance genes. Moreover, aminoglycoside resistance and resistance genes were found to be transferable in most strains (24/37). Nevertheless, pulsed-field gel electrophoresis (PFGE) and dendrogram analysis showed clonal diversity among these isolates. S1 nuclease PFGE, Southern hybridization, and whole-genome sequencing indicated that aac(3)-IIg was located on transferable as well as nontransferable plasmids of various sizes. The analysis of the genetic environment suggested that aac(3)-IIg is embedded within a class 1 integron, with IS26 playing an important role in its mobility.


Test-retest reliability in fMRI: Or how I learned to stop worrying and love the variability.
This paper briefly recounts the details of one of the first studies of whole brain, single subject variability in functional Magnetic Resonance Imaging (fMRI). In this paper I will not attempt to provide an overview of fMRI reliability, but will instead revisit the choices we made in performing our experiment as we did.


Organizational determinants of patient-centered fertility care: a multilevel analysis.
To identify organizational determinants of positive patient experiences with fertility care, with the goal of improving patient centeredness of care. Cross-sectional survey. One large university clinic and 12 medium-sized fertility clinics in the Netherlands. Three hundred and sixty-nine couples receiving medically assisted reproduction in one of the participating clinics between March and May 2008. None. Organizational determinants of patients' experiences with patient centeredness in fertility care. Of the patients during the relevant period, 78% of the women and 76% of their partners participated in the study. Infertile couples who have a lead physician, have access to an electronic personal health record, or see trained fertility nurses have more positive experiences with aspects of patient-centered care, like continuity of care and partner involvement. Moreover, receiving a treatment other than in vitro fertilization was negatively associated with the perceived patient centeredness of care. The identified determinants explained 5.1% to 22.4% of the total variance. This study provides organizational determinants of patients' experiences with fertility care on numerous facets of patient centeredness. These organizational determinants can be used as valuable tools to enable clinics to provide a more positive patient experience.


Erratum: Filip, S.; et al. Therapeutic Apheresis, Circulating PLD, and Mucocutaneous Toxicity: Our Clinical Experience through Four Years. Pharmaceutics 2020, 12, 940.
The following correction has been made to this paper [...].


G protein activation: a receptor-independent mode of action for cationic amphiphilic neuropeptides and venom peptides.
The neuropeptide substance P, the venom peptide mastoparan and the synthetic polyamine compound 48/80 activate rat peritoneal mast cells, leading to rapid histamine release by exocytosis. Although these effects are inhibited by pertussis toxin and involve a transient increase in IP3, no selective membrane receptors have been identified. However, it has recently been shown that these compounds activate G proteins in vitro. Here Yves Landry and colleagues discuss the proposal that direct activation of G protein is the physiological mechanism of action of substance P on rat peritoneal mast cells, this mechanism being mimicked by mastoparan and 48/80, and possibly by other cationic amphiphilic peptides such as kinins. These compounds might be of help in defining the interaction between membrane receptors and G proteins.


Comparative study of computed tomography (CT) and carotid angiography (CAG) in stroke patients.
The findings in CT and CAG were compared in 173 stroke patients to evaluate dependability in the localization of cerebral arterial occlusion and of infarction by CT, and to test the reliability of the angiographic and clinical diagnosis of a space-occupying lesion in patients diagnosed clinically as stroke cases.


Protein synthesis in rabbit reticulocytes. Purification and properties of an Mr 80,000 polypeptide (Co-eIF-2A80) with Co-eIF-2A activity.
The high molecular weight protein complex, Co-eIF-2, contains both Co-eIF-2A and Co-eIF-2C activities (Bagchi, M. K., Banerjee, A. C., Roy, R., Chakravarty, I., and Gupta, N. K. (1982) Nucleic Acids Res. 10, 6501-6510). Co-eIF-2A stimulated Met-tRNAf binding to eukaryotic initiation factor-2 (eIF-2) both in the presence and absence of Mg2+. Co-eIF-2C stimulates Met-tRNAf binding to eIF-2 in the presence of Mg2+ by relieving Mg2+ inhibition of ternary complex formation from eIF-2. Co-eIF-2 protein complex contains several polypeptides including Mr 80,000 and 50,000 polypeptides. Three polypeptides (Mr 80,000, 50,000 and 25,000) are present in 0.5 M KCl ribosomal salt wash and each possesses Co-eIF-2A activity. Mr 80,000 polypeptide (Co-eIF-2A80) has been purified to homogeneity and its properties studied. 1) Co-eIF-2A80 stimulated Met-tRNAf binding to eIF-2 and the complex formed was resistant to aurintricarboxylic acid. 2) Co-eIF-2A80 activity was N-ethylmaleimide-resistant and heat-labile; it was destroyed by heating at 55 degrees C for 4 min. 3) Antibodies prepared against homogeneous Co-eIF-2A80 strongly inhibited protein synthesis in reticulocyte lysates and, also, eIF-2 and Co-eIF-2 promoted Met-tRNAf binding to 40 S ribosomes. Inhibition of protein synthesis in reticulocyte lysates was overcome by preincubation of anti-Co-eIF-2A80 with homogeneous Co-eIF-2A80 and was partially overcome by similar preincubation with Co-eIF-2. 4) Upon limited digestion with Staphylococcus aureus V8 protease, the homogeneous Co-eIF-2A80 gave two major polypeptide fragments (Mr 50,000 and 25,000). Upon similar treatment, an Mr 80,000 polypeptide band isolated from the sodium dodecyl sulfate-gel of the Co-eIF-2 protein complex gave four major polypeptide fragments, and two of these fragments (Mr 50,000 and 25,000) were similar to those given by Co-eIF-2A80, indicating that this Mr 80,000 polypeptide band contains the Co-eIF-2A80 component. We suggest that Co-eIF-2A80 is a component of Co-eIF-2 and is also essential for Co-eIF-2 activity and overall peptide chain initiation.


Abnormal glycosylation and altered Golgi structure in colorectal cancer: dependence on intra-Golgi pH.
Abnormal glycosylation of cellular glycoconjugates is a common phenotypic change in many human tumors. Here, we explore the possibility that an altered Golgi pH may also be responsible for these cancer-associated glycosylation abnormalities. We show that a mere dissipation of the acidic Golgi pH results both in increased expression of some cancer-associated carbohydrate antigens and in structural disorganization of the Golgi apparatus in otherwise normally glycosylating cells. pH dependence of these alterations was confirmed by showing that an acidification-defective breast cancer cell line (MCF-7) also displayed a fragmented Golgi apparatus, whereas the Golgi apparatus was structurally normal in its acidification-competent subline (MCF-7/AdrR). Acidification competence was also found to rescue normal glycosylation potential in MCF-7/AdrR cells. Finally, we show that abnormal glycosylation is also accompanied by similar structural disorganization and fragmentation of the Golgi apparatus in colorectal cancer cells in vitro and in vivo. These results suggest that an inappropriate Golgi pH may indeed be responsible for the abnormal Golgi structure and lowered glycosylation potential of the Golgi apparatus in malignant cells.


Formulas for the quantitation of intrathecal IgG production. Their validity in the presence of blood-brain barrier damage and their utility in multiple sclerosis.
There are several formulas for the quantitative determination of intrathecal IgG production: Reiber and Felgenhauer's formula (IgG(loc)), the Extended IgG index, Tourtellotte's formula (TOURT), Schuller and Sagar's formula (SCHULL), the IgG index, the Log IgG index, and Blennow and co-workers' formula (IGGPROD). To evaluate the utility of these formulas in the presence of blood-brain barrier (BBB) damage, we present the results from a study of serum and cerebrospinal fluid (CSF) samples from 125 healthy individuals, 18-88 years of age; 1072 consecutive patients without oligoclonal IgG bands (OCBs) in the CSF, 683 without BBB damage (CSF/S: albumin ratio < 9.8) and 389 with BBB damage (CSF/S albumin ratio 9.8-30); and 106 patients with definite multiple sclerosis (MS). The relation between the CSF/S albumin ratio and the CSF/S IgG ratio was remarkably linear in both healthy individuals (r = 0.95; P < 0.0001) and patients without oligoclonal bands in the CSF (r = 0.95; P < 0.0001). Therefore, IgG(loc) and the Extended IgG index, two formulas based on a nonlinear relation between the CSF/S albumin ratio and the CSF/S IgG ratio, yielded biased results (lower values) in the presence of BBB damage. TOURT and SCHULL also yielded biased (higher) values in the presence of BBB damage, probably because of incorrect constants in these formulas. There were no significant correlations between the CSF/S albumin ratio (i.e. the BBB function) and the IgG index or the Log IgG index, two dimensionless quotients for the detection of intrathecal IgG production, or between the CSF/S albumin ratio and IGGPROD, an empirical formula for the determination of intrathecal IgG production in mg/l.(ABSTRACT TRUNCATED AT 250 WORDS)


The Associations Between Workplace Noise, Ototoxic Chemicals, and Tinnitus.
To investigate the occupational risk factors associated with tinnitus in the Australian working population. The research was conducted using data collected from the Australian Workplace Exposure Survey-Hearing, a national cross-sectional study of 4970 workers conducted in 2016 to 2017. Workers were asked if they experienced tinnitus and if they answered affirmatively, they were asked about the frequency and length of the presentations. Based on their answers, each worker was categorized as having no tinnitus or any tinnitus, with an additional group of workers with any tinnitus subcategorized as having constant tinnitus. Exposure assessment was conducted using an automated expert assessment method. Exposures included daily noise (L Aeq,8h ), hand-arm vibration (A(8)), impulse noise, smoking status, styrene, trichloroethylene, toluene, n-hexane, p-xylene, ethylbenzene, lead, and carbon monoxide. Univariate and multivariate logistic regression models were used to examine the associations between workplace exposures and tinnitus. Workers with an estimated noise exposure above the workplace limit (L Aeq,8h > 85 dBA) had 1.73 (95% confidence interval [CI]: 1.42 to 2.11) increased odds of any tinnitus and 2.15 (95% CI: 1.60 to 2.89) odds of constant tinnitus. The odds of workers having any tinnitus increased with increasing noise exposure levels in a dose-response relationship that strengthened when considering only those with constant tinnitus. In the fully adjusted model, statistically significant associations were seen for lead exposure with both any and constant tinnitus, toluene exposure with constant tinnitus, and carbon monoxide exposure with any tinnitus. These results suggest that, in addition to workplace noise, occupational exposures to lead, toluene, and carbon monoxide are associated with tinnitus.


Toxic and developmental effects of organophosphorus insecticides in embryos of the South African clawed frog.
Four organophosphorus insecticides and the active metabolites of two phosphorothionate insecticides were tested for their toxic and teratogenic effects on embryos of Xenopus laevis. All compounds caused dose-dependent developmental defects, such as abnormal pigmentation, abnormal gut development, notochordal defects and reduced growth. Malathion, malaoxon, parathion, and paraoxon produced severe defects, while monocrotophos and dicrotophos produced considerably milder defects. All compounds reduced NAD+ levels to a similar extent, regardless of the severity of the defects induced. Thus some commonly used organophosphorus insecticides and their metabolites are teratogenic to Xenopus embryos, but reduced NAD+ level does not appear to be important in causing the developmental defects.


The combined associations of social participation and support with self-rated health and dietary satisfaction in men with spinal cord injury.
Cross-sectional study. (1) To examine the association between social participation (SP) and social support (SS) with self-rated health and dietary satisfaction and (2) to explore the joint association and interactions of SP and SS with self-rated health and dietary satisfaction in community-dwelling adult men with spinal cord injury. Members of the Spinal Injuries Japan organization. We sent questionnaires to 2731 registered members of Spinal Injuries Japan via mail. Responses from 625 men aged ⩾40 years were analyzed. Respondents were categorized into four groups: SP/sufficient SS, SP/insufficient SS, no SP/sufficient SS and no SP/insufficient SS. Logistic regression analysis was used to examine the odds ratios for self-rated health and dietary satisfaction according to the SP/SS categories. Relative to participants in the no SP/insufficient SS category, those in the SP/sufficient SS group demonstrated significantly better self-rated health and dietary satisfaction after adjusting for sociodemographic variables. There was no interaction between SP and SS in self-rated health or dietary satisfaction. SP was associated with high self-rated health without SS, and sufficient SS was associated with high dietary satisfaction without SP. Relative to other groups, participants with SP/sufficient SS demonstrated higher self-rated health and dietary satisfaction. Sufficient SS was associated with high dietary satisfaction without SP. This study suggested the importance of addressing aspects of both SP and SS using self-rated health and dietary satisfaction as outcome measures in health promotion programs.


The effect of preoperative keratometry on visual outcomes after moderate myopic LASIK.
To evaluate visual outcomes in moderately myopic eyes with flat and steep corneas (preoperatively) that have been treated with laser-assisted in situ keratomileusis (LASIK). Records of ninety-six eyes with average preoperative keratometry (K) values between 39.9 and 42.0 diopters (D) (flat) were matched with 103 eyes with preoperative K values between 46.0 and 47.2 D (steep) that underwent LASIK between March 2007 and March 2010 for moderate myopia, and were retrospectively reviewed. The primary outcome measures used to determine the effect of preoperative keratometry on visual prognosis were refraction, visual acuity, change in keratometry (ΔK), and change in spherical equivalent (ΔSE), measured at 1, 3, 6, and 12 months postoperatively. Significant differences were found at 6 months postoperatively between the flat group and steep group in SE (P = 0.029), sphere (P = 0.018), ΔK (P = 0.002), percentage of eyes achieving SE of -0.25 to + 0.25 D (P = 0.0125), -0.26 to -0.50 D (P = 0.003), -0.51 to -1.00 D (P = 0.044), and the percentage of eyes achieving uncorrected distance visual acuity of 20/15 or better (P = 0.0006). Moderately myopic eyes with flatter corneas preoperatively have better visual prognosis following LASIK compared with moderately myopic eyes with steeper corneas.


Acute right ventricular infarction without infero-posterior left ventricular infarction.
Two cases of acute right ventricular infarction associated with acute extensive anterior myocardial infarction in the absence of inferior and/or posterior left ventricular infarction are presented. Such a combination is likely to occur from acute occlusion of the left anterior descending artery in the face of severe narrowing of the infundibular (conus) artery rather than from acute occlusion of the right coronary artery.


Alpinia officinarum mediated copper oxide nanoparticles: synthesis and its antifungal activity against Colletotrichum gloeosporioides.
Green synthesis offers an environmentally friendly and cost-effective alternative for the synthesis of copper oxide nanoparticles (CuO NPs). In this study, the synthesis of CuO NPs was optimized by using copper sulfate (CuSO4) and the aqueous extract of Alpinia officinarum and its antifungal activity were investigated. The synthesized CuO NPs were characterized by UV-visible spectroscopy (UV-vis), X-ray diffraction (XRD), Fourier-transform infrared radiation spectroscopy (FT-IR), scanning electron microscope (SEM), energy dispersive spectroscopy (EDS), dynamic light scattering (DLS), and transmission electron microscopy (TEM). The results showed that the optimized conditions for the synthesis of CuO NPs were 1:2 ratio of extract and CuSO4 solution, pH 7, and 30 °C. The characteristic UV-vis peak of A. officinarum synthesized CuO NPs was at 264 nm. The synthesized CuO NPs had high crystallinity and purity and were spherical in morphology with the mean size of 46.40 nm. The synthesized CuO NPs reduced the fungal growth of Colletotrichum gloeosporioides in a dose-dependent manner. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of the CuO NPs were 125 μg·mL-1 and 500 μg·mL-1, respectively. The antifungal activity of CuO NPs may be attributed to its ability to deform the structure of fungal hyphae, induce excessive reactive oxygen species accumulation and lipid peroxidation in fungi, disrupt the mycelium cell membrane, and result cellular leakage.


Student nurses experience of a "fairy garden" healing haven garden for sick children.
The concept and philosophy of healing environments in health care is not new and there has been recent research into the experience of nurses and families experience of healing environments producing positive outcomes in relieving stress and improving quality of life. However, there is little in-depth information about student nurse's experience of healing environments in support of patients. To report on the stories of student nurses who participated in formal and informal activities in a healing haven environment called a Fairy Garden (FG) within a hospital in northern Thailand. Their beliefs about the care of sick children in an environment designed to provide educational and recreational activity during hospital care are explored. Narrative inquiry, a qualitative methodology was selected to capture the main threads of the participants' experience. Clandinin's narrative inquiry framework involving the three commonality dimensions of sociality, temporality and place were used in analysing the data. Sixty-two student nurses from a Thai College of Nursing and from an Australian university were interviewed. In this study the place of a FG has been investigated as a non-clinical environment providing sick children with exposure to nature, play activities and spaces to explore. Findings include three main threads: freedom to be a child not a sick child, engaging in care and professionalism, a moment in time of living fantasy. Student nurses in this study had a broader understanding of health care other than the biomedical model. It transformed their learning and opened their eyes to a more holistic approach to humanising care of sick children.


The rabbit C family of short, interspersed repeats. Nucleotide sequence determination and transcriptional analysis.
The C repeat family was first observed in the rabbit beta-like gene family. We have estimated the repetition frequency of the C repeats, determined the nucleotide sequence of three intact members and one truncated member, and have investigated the size, tissue specificity, and intracellular localization of C repeat transcripts. Members of the C repeat family are short (average size of 316 base-pairs) and are repeated about 170,000 times per haploid genome in a widely dispersed pattern. They end in a 3' poly(dA) tract and are flanked by direct repeats that range in size from 8 to 16 base-pairs. The consensus internal control regions for polymerase III transcription are located near the 5' end. Different amounts of C repeat RNA accumulate in a variety of tissues, and most of the transcripts are confined to the nucleus. A heterogeneous distribution of C repeat RNA sizes was found, ranging from about 330 to 8200 nucleotides. These structural and transcriptional properties are similar to those of primate and rodent Alu and Alu-like repeats. However, the C repeats are not similar in sequence to the Alu repeats. Thus two different types of short, interspersed repeats capable of being transcribed and proposed to be transposable elements have now been identified in mammals. The positions of these short repeats in mammalian beta-like globin gene families are not tightly conserved.


Effect of the NAD(P)H oxidase inhibitor, apocynin, on peripheral nerve perfusion and function in diabetic rats.
Upregulation of vascular NAD(P)H oxidase has been considered an important source for elevated levels of reactive oxygen species that contribute to several cardiovascular disease states, including the vascular complications of diabetes mellitus. Previous studies have shown that treatment with antioxidants corrects impaired nerve function and blood flow in diabetic rats. The aim was to assess the degree of involvement of NAD(P)H oxidase in experimental diabetic neuropathy. To this end, after 6 weeks of untreated streptozotocin-diabetes, rats were treated for 2 weeks with the NAD(P)H oxidase, apocynin. Two high doses (15 and 100 mg/kg) were used to ensure that maximal effects were registered. Diabetes caused a 20% reduction in sciatic nerve motor conduction velocity, and a 14% deficit for sensory saphenous nerve. Apocynin treatment corrected these defects by 32% and 48%, respectively: there were no significant differences between the effects of the 2 doses. Sciatic nerve nutritive endoneurial perfusion was measured by hydrogen clearance microelectrode polarography. Blood flow and vascular conductance were 47% and 40% reduced by diabetes, respectively. Both doses of apocynin had similar effects, correcting the blood flow deficit by 31% and conductance by 47%. Thus, the data show that NAD(P)H oxidase contributes to the neurovascular deficits in diabetic rats. While only accounting for part of the elevated reactive oxygen species production in diabetes, this mechanism could provide a novel therapeutic candidate for further investigation in diabetic neuropathy and vasculopathy.


Morphological and biochemical changes in the gastric mucosa of A/HEJ mice injected with a xenogeneic stomach antigen.
Subcutaneous injection of W/Fu rat stomach antigen emulsified with an equal volume of Freund's complete adjuvant at 1 week interval for a total of 2 administrations induced atrophic gastritis in A/HeJ mice. From the second week after the last injection the parietal cell mass decreased. At the same time, Alcian blue-PAS positive cells and Paneth-like cells appeared. Furthermore, the activity of alkaline phosphatase and disaccharidase of the small intestinal mucosa was biochemically demonstrated in the atrophic glandular stomach.