Efficiency improvements of x-ray simulations in EGSnrc user-codes using bremsstrahlung cross-section enhancement (BCSE).
This paper presents the implementation of the bremsstrahlung cross-section enhancement (BCSE) variance-reduction technique into the EGSnrc/BEAMnrc system. BCSE makes the simulation of x-ray production from bremsstrahlung targets more efficient; it does so by artificially making the rare event of bremsstrahlung emission more abundant, which increases the number of statistically-independent photons that contribute to reducing the variance of the quantity of interest without increasing the CPU time appreciably. BCSE does not perturb the charged-particle transport in EGSnrc and it is made compatible with all other variance-reduction techniques already used in EGSnrc and BEAMnrc, including range rejection, uniform bremsstrahlung splitting, and directional bremsstrahlung splitting. When optimally combining BCSE with splitting to simulate typical situations of interest in medical physics research and in clinical practice, efficiencies can be up to five orders of magnitude larger than those obtained with analog simulations, and up to a full order of magnitude larger than those obtained with optimized splitting alone (which is the state-of-the-art of the EGSnrc/BEAMnrc system before this study was carried out). This study recommends that BCSE be combined with the existing splitting techniques for all EGSnrc/BEAMnrc simulations that involve bremsstrahlung targets, both in the kilovoltage and megavoltage range. Optimum crosssection enhancement factors for typical situations in diagnostic x-ray imaging and in radiotherapy are recommended, along with an easy algorithm for simulation optimization.


The effect of turning and backrub on mixed venous oxygen saturation in critically ill patients.
To examine the effect of a change in body position (right or left lateral) and timing of backrub (immediate or delayed) on mixed venous oxygen saturation in surgical ICU patients. A repeated-measures design was used to study 57 critically ill men. Mixed venous oxygen saturation was recorded at 1-minute intervals for 5 minutes in each of three periods: baseline, after turning, and after backrub. Subjects were randomly assigned to body position and timing of backrub. Subjects in the immediate-backrub group were turned and given a 1-minute backrub. Mixed venous oxygen saturation was measured at 1-minute intervals for 5 minutes at two points: after the backrub and then with the patient lying on his side. For subjects in the delayed-backrub group, saturation was measured at 1-minute intervals for 5 minutes at two different points: after the subject was turned to his side and after the backrub. Both position and timing of backrub had significant effects on mixed venous oxygen saturation across conditions over time. Subjects positioned on their left side had a significantly greater decrease in saturation when the backrub was started. At the end of the backrub, saturation was significantly lower in subjects lying on their left side than in subjects lying on their right side. The pattern of change differed according to the timing of the backrub, and return to baseline levels of saturation after intervention differed according to body position. Two consecutive interventions (change in body position and backrub) cause a greater decrease in mixed venous oxygen saturation than the two interventions separated by a 5-minute equilibration period. Turning to the left side decreases oxygen saturation more than turning to the ride side does. Oxygen saturation returns to clinically acceptable ranges within 5 minutes of an intervention. In patients with stable hemodynamic conditions, the standard practice of turning the patient and immediately giving a backrub is recommended. However, it is prudent to closely monitor individual patterns of mixed venous oxygen saturation, particularly in patients with unstable hemodynamic conditions.


Ameliorating diabetes-associated atherosclerosis and diabetic nephropathy through modulation of soluble guanylate cyclase.
Diabetes mellitus (DM) is an independent risk factor for micro- and macrovascular complications such as nephropathy and atherosclerosis respectively, which are the major causes of premature morbidity and mortality in Type 1 and Type 2 diabetic patients. Endothelial dysfunction is the critical first step of vascular disease and is characterized by reduced bioavailability of the essential endothelial vasodilator, nitric oxide (NO), coupled with an elevation in inflammation and oxidative stress. A novel pathway to bolster NO activity is to upregulate soluble guanylate cyclase (sGC), an enzyme responsible for mediating the protective actions of NO. Two classes of sGC modulators exist, activators and stimulators, with differing sensitivity to oxidative stress. In this study, we investigated the therapeutic effects of the sGC stimulator BAY 41-2272 (Bay 41) and the sGC activator BAY 60-2770 (Bay 60) on endpoints of atherosclerosis and renal disease as well as inflammation and oxidative stress in diabetic Apolipoprotein E knockout (ApoE-/-) mice. We hypothesized that under oxidative conditions known to accompany diabetes, sGC activation might be more efficacious than sGC stimulation in limiting diabetic vascular complications. We demonstrate that Bay 60 not only significantly decreased nitrotyrosine staining (P < 0.01) and F4/80 positive cells by 75% (P < 0.05), but it also significantly reduced total plaque area (P < 0.05) and improved endothelial function (P < 0.01). Our data suggest an important anti-atherogenic role for Bay 60 accompanied by reduced oxidative stress and inflammation under diabetic settings. Treatment with the stimulator Bay 41, on the other hand, had minimal effects or caused no changes with respect to cardiovascular or renal pathology. In the kidneys, treatment with Bay 60 significantly lessened urinary albuminuria, mesangial expansion and nitrotyrosine staining under diabetic conditions. In summary, our head-to-head comparator is the first preclinical study to show that a sGC activator is more efficacious than a sGC stimulator for the treatment of diabetes-associated vascular and renal complications.


Enhancing physiologic simulations using supervised learning on coarse mesh solutions.
Computational modelling of physical and biochemical processes has emerged as a means of evaluating medical devices, offering new insights that explain current performance, inform future designs and even enable personalized use. Yet resource limitations force one to compromise with reduced order computational models and idealized assumptions that yield either qualitative descriptions or approximate, quantitative solutions to problems of interest. Considering endovascular drug delivery as an exemplary scenario, we used a supervised machine learning framework to process data generated from low fidelity coarse meshes and predict high fidelity solutions on refined mesh configurations. We considered two models simulating drug delivery to the arterial wall: (i) two-dimensional drug-coated balloons and (ii) three-dimensional drug-eluting stents. Simulations were performed on computational mesh configurations of increasing density. Supervised learners based on Gaussian process modelling were constructed from combinations of coarse mesh setting solutions of drug concentrations and nearest neighbourhood distance information as inputs, and higher fidelity mesh solutions as outputs. These learners were then used as computationally inexpensive surrogates to extend predictions using low fidelity information to higher levels of mesh refinement. The cross-validated, supervised learner-based predictions improved fidelity as compared with computational simulations performed at coarse level meshes--a result consistent across all outputs and computational models considered. Supervised learning on coarse mesh solutions can augment traditional physics-based modelling of complex physiologic phenomena. By obtaining efficient solutions at a fraction of the computational cost, this framework has the potential to transform how modelling approaches can be applied in the evaluation of medical technologies and their real-time administration in an increasingly personalized fashion.


Diabetes responsive to intravenous but not subcutaneous insulin: effectiveness of aprotinin.
Patients with diabetes that is insensitive to subcutaneous insulin but sensitive to intravenous insulin have recently been described. We have studied this phenomenon is five female diabetics (14 to 31 years of age) who required excessive amounts of insulin (2.5 to 30.0 units per kilogram of body weight per day) to avoid recurrent ketoacidosis. Known causes of insulin resistance were excluded. All patients had normal responses to conventional doses of intravenous insulin (0.35 to 0.9 unit per kilogram per day). Four patients required continuous intravenous infusion of insulin for one to six months. When a mixture of aprotinin (a protease inhibitor) and regular porcine insulin was given subcutaneously, conventional doses (0.7 to 1.4 units per kilogram per day) produced euglycemia; plasma levels of free insulin rose, and ketonuria disappeared. Four patients had episodes of spontaneous, severe hypoglycemia before and during aprotinin therapy, necessitating continuous infusion of glucose for two to 14 days. Although no insulin was administered, hyperinsulinemia (50 to 2000 muU of free insulin per milliliter [359 to 14,350 pmol per liter]) was present. These findings suggest excessive degradation or sequestration of insulin at the site of injection.


Blank comparison analysis of emergent symbolic mapping by young children.
Eight typically developing children first learned to select pictures in response to dictated words in a three-choice matching-to-sample format. Next, the matching-to sample baseline was transformed via a stimulus control shaping procedure: Matching trials displayed two pictures and a third stimulus (termed a "blank" comparison). The blank was to be selected if the dictated sample did not match either of the displayed pictures. The blank comparison baseline was then used to evaluate the children's response to matching displays that presented novel sample and comparison stimuli. The blank comparison method permitted the children to indicate whether the novel sample-comparison pair were related (by selecting the picture) or not related (by selecting the blank). The method was used to evaluate emergent symbolic mapping and learning of new word:picture matching relations by the children. This study illustrates new, potentially more informative methods for analyzing symbol mapping in young children. The methods were shown to have considerable promise for advancing theoretical analyses of emergent mapping in both behavior analytic and developmental language research.


A microbiological study of ophthalmia neonatorum in hospital-born babies.
A retrospective microbiological analysis of ophthalmia neonatorum among the hospital-born babies of a rural multispecialty hospital over a period of 2 years was made. Diagnostic cultures were performed in all the affected babies and 30 cases were found to be culture positive for bacterial species. The spectrum of various bacterial isolates and their antibiotic sensitivity patterns were studied. Pseudomonas aeruginosa was found to be the commonest causative organism according to this study and newer fluoroquinolones like ciprofloxacin and norfloxacin were the antibiotics that showed the best sensitivity pattern.


How Histone Deacetylase Inhibitors Alter the Secondary Metabolites of Botryosphaeria mamane, an Endophytic Fungus Isolated from Bixa orellana.
Fungi are talented organisms able to produce several natural products with a wide range of structural and pharmacological activities. The conventional fungal cultivation used in laboratories is too poor to mimic the natural habitats of fungi, and this can partially explain why most of the genes responsible for the production of metabolites are transcriptionally silenced. The use of Histone Deacetylase inhibitors (HDACis) to perturb fungal secondary biosynthetic machinery has proven to be an effective approach for discovering new fungal natural products. The present study relates the effects of suberoylanilide hydroxamic acid (SAHA) and sodium valproate (VS) on the metabolome of Botryosphaeria mamane, an endophytic fungus isolated from Bixa orellana L. UHPLC/HR-MS analysis, integrated with four metabolomics tools: MS-DIAL, MS-FINDER, MetaboAnalyst and GNPS molecular networking, was established. This study highlighted that SAHA and VS changed metabolites in B. mamane, causing upregulation and downregulation of metabolites production. In addition, twelve compounds were detected in the extracts as metabolites structurally correlated to SAHA, indicating its important reactivity in the medium or its metabolism by the fungus. An addition of SAHA induced the production of eight metabolites while VS induced only two metabolites undetected in the control strain. This result illustrates the importance of adding HDACis to a fungal culture in order to induce metabolite production.


Interaction and co-encapsidation of human immunodeficiency virus type 1 Gag and Vif recombinant proteins.
Human immunodeficiency virus type 1 (HIV-1) wild-type (WT) virion infectivity factor (Vif) protein (Vifwt) and full-length Gag precursor (Pr55Gag) were found to be co-encapsidated into extracellular, membrane-enveloped virus-like particles released by budding from Sf9 cells co-expressing the two recombinant proteins in trans, with an average copy number of 3.5+/-0.6 Vifwt per 100 Pr55Gag molecules. No preferential localization at the plasma membrane was observed for recombinant Vif in the absence of Gag expression, and a significant proportion of Vif accumulated within the nucleus. Two conserved motifs, W89RKRRY94 and P156KKIKP161, seemed to act as nuclear addressing signals. The Pr55Gag and Vifwt interacting domains were analysed by biopanning of a phage-displayed hexapeptide library. The Vif-binding domain, which spanned residues H421-T470 in Pr55Gag, corresponded to the C-terminal region of nucleocapsid (NC), including the second zinc finger, the intermediate spacer peptide sp2 and the N-terminal half of the p6 domain. Deletions in these Gag domains significantly decreased the Vif encapsidation efficiency, and complete deletion of NC abolished Vif encapsidation. In Vif, four discrete Gag-binding sites were identified, within residues T68-L81 (site I) and W89-P100 (site II) in the central domain, and within residues P162-R173 (III) and P177-M189 (IV) at the C terminus. Substitutions in site I and deletion of site IV were detrimental to Vif encapsidation, whereas substitution of basic residues for alanine in sites III and IV had a positive effect. The data suggest a direct intracellular Gag-Vif interaction and the occurrence of a Pr55Gag-mediated membrane-targeting pathway for Vif in Sf9 cells.


Influence of antiseptic lavage during tibial plateau leveling osteotomies on surgical site infection in 1422 dogs.
To determine the influence of preclosure antiseptic versus saline lavage on surgical site infections (SSI) in dogs following tibial plateau leveling osteotomy (TPLO). A multicenter retrospective study. Dogs treated with TPLO (n = 1422) between December 2019 and October 2021. The medical records of dogs treated with TPLO were reviewed for preclosure antiseptic lavage or saline irrigation. Additional collected data included signalment, unilateral or bilateral TPLO, postoperative administration of antimicrobial medications, antibiotic agent, surgeon, and development of SSI within 90 postoperative days. Eleven factors were analyzed for association with SSI using univariate and multivariate analysis. Data were collected from the records of 519 dogs treated with antiseptic lavage and 903 dogs treated with saline lavage during TPLO. Surgical site infections were diagnosed more frequently in dogs that received preclosure antiseptic lavage (77/519, 14.84%) than those with saline irrigation (80/903, 8.86%) (p = .001). Single session bilateral TPLO increased the odds of SSI by 2.5x (p = .004). The odds of SSI increased by 11% (p = .001) for every 5 kg increase in bodyweight. Postoperative administration of antimicrobials decreased the risk of SSI (p = .008). The preclosure antiseptic lavage tested here did not decrease the incidence of SSI after TPLO. The results of this study do not provide evidence to support preclosure antiseptic lavage during TPLOs.


Vaccination against Lyme borreliosis.
Lyme borreliosis is a worldwide family of tick-borne infections caused by the spirochete Borrelia burgdorferi sensu lato. It is the most common tick-borne human infection in the Western world. There are several subgroups of the spirochete. Two monovalent vaccines against this infection have been presented in the USA, both of which use the borrelial outer surface protein A (OspA) as antigen. The first of these vaccines has been released for general use. A European polyvalent vaccine using the antigen OspC is undergoing clinical trial in the Aland Islands in Finland. Lately, another antigen group, decorin-binding proteins (Dbp), has been considered for immunization purposes. A European vaccine must be effective against several subgroups of the borrelia spirochete, and this complicates the situation compared with that in the USA, where one spirochete subspecies dominates the scene.


Dynamics of a methanol-fed marine denitrifying biofilm: 1-Impact of environmental changes on the denitrification and the co-occurrence of Methylophaga nitratireducenticrescens and Hyphomicrobium nitrativorans.
The biofilm of a methanol-fed denitrification system that treated a marine effluent is composed of multi-species microorganisms, among which Hyphomicrobium nitrativorans strain NL23 and Methylophaga nitratireducenticrescens strain JAM1 are the principal bacteria involved in the denitrifying activities. Here, we report the capacity of the denitrifying biofilm to sustain environmental changes, and the impact of these changes on the co-occurrence of H. nitrativorans and M. nitratireducenticrescens. In a first set of assays, the original biofilm (OB) was cultivated in an artificial seawater (ASW) medium under anoxic conditions to colonize new carriers. The new formed biofilm was then subjected to short exposures (1-5 days) of a range of NaCl, methanol, nitrate (NO3 -) and nitrite (NO2 -) concentrations, and to different pHs and temperatures. In a second set of assays, the OB was cultivated in ASW medium for five weeks with (i) a range of NaCl concentrations, (ii) four combinations of NO3 -/methanol concentrations and temperatures, (iii) NO2 -, and (iv) under oxic conditions. Finally, the OB was cultivated for five weeks in the commercial Instant Ocean (IO) seawater. The growth of the biofilm and the dynamics of NO3 - and NO2 - were determined. The levels of M. nitratireducenticrescens and H. nitrativorans were measured by qPCR. In the first set of assays, the biofilm cultures had the capacity to sustain denitrifying activities in most of the tested conditions. Inhibition occurred when they were exposed to high pH (10) or to high methanol concentration (1.5%). In the second set of assays, the highest specific denitrification rates occurred with the biofilm cultures cultivated at 64.3 mM NO3 - and 0.45% methanol, and at 30 °C. Poor biofilm development occurred with the biofilm cultures cultivated at 5% and 8% NaCl. In all biofilm cultures cultivated in ASW at 2.75% NaCl, H. nitrativorans strain NL23 decreased by three orders of magnitude in concentrations compared to that found in OB. This decrease coincided with the increase of the same magnitude of a subpopulation of M. nitratireducenticrescens (strain GP59 as representative). In the biofilm cultures cultivated at low NaCl concentrations (0% to 1.0%), persistence of H. nitrativorans strain NL23 was observed, with the gradual increase in concentrations of M. nitratireducenticrescens strain GP59. High levels of H. nitrativorans strain NL23 were found in the IO biofilm cultures. The concentrations of M. nitratireducenticrescens strain JAM1 were lower in most of the biofilms cultures than in OB. These results demonstrate the plasticity of the marine methylotrophic denitrifying biofilm in adapting to different environmental changes. The NaCl concentration is a crucial factor in the dynamics of H. nitrativorans strain NL23, for which growth was impaired above 1% NaCl in the ASW-based biofilm cultures in favor of M. nitratireducenticrescens strain GP59.


Disparities in the Diagnosis and Management of Infants Hospitalized With Inadequate Weight Gain.
To evaluate the association between race and the named etiology for inadequate weight gain among hospitalized infants and assess the differences in management. This single-center retrospective cohort study of infants hospitalized for the workup and management of inadequate weight gain used infant race and neighborhood-level socioeconomic deprivation as exposures. The etiology of inadequate weight gain was categorized as nonorganic, subjective organic (ie, gastroesophageal reflux and cow's milk protein intolerance), or objective organic (eg, hypothyroidism). The management of inadequate weight gain was examined in secondary outcomes. Among 380 infants, most were white and had a nonorganic etiology of inadequate weight gain. Black infants had 2.3 times higher unadjusted odds (95% credible interval [CI] 1.17-4.76) of a nonorganic etiology of inadequate weight gain compared with white infants. After adjustment, there was no association between race and etiology (adjusted odds ratio 0.8, 95% CI [0.44-2.08]); however, each 0.1 increase in neighborhood-level deprivation was associated with 80% increased adjusted odds of a nonorganic etiology of inadequate weight gain (95% CI [1.37-2.4]). Infants with a nonorganic etiology of inadequate weight gain were more likely to have social work and child protective service involvement and less likely to have nasogastric tube placement, gastroenterology consults, and speech therapy consults. Infants from neighborhoods with greater socioeconomic deprivation were more likely to have nonorganic causes of inadequate weight gain, disproportionately affecting infants of Black race. A nonorganic etiology was associated with a higher likelihood of social interventions and a lower likelihood of medical interventions.


Variation of glucosinolate accumulation and gene expression of transcription factors at different stages of Chinese cabbage seedlings under light and dark conditions.
Chinese cabbage is one of the most important leafy vegetables widely used in East Asian cuisines. The glucosinolate (GSL) accumulation and transcript levels of 7 transcription factors (Dof1.1, IQD1-1, MYB28, MYB29, MYB34, MYB51, and MYB122, and their isoforms) involved in the biosynthesis of aliphatic and indolic glucosinolates (GSLs) were analyzed at different stages of Chinese cabbage (Brassica rapa ssp. pekinensis) seedlings under light and dark conditions using high performance liquid chromatography and quantitative real time PCR. During seedling development, transcription of almost all transcription factors under light conditions was higher expressed than under dark conditions. Five aliphatic GSLs (progoitrin, sinigrin, glucoalyssin, gluconapin, and glucobrassicanapin) and four indolic GSLs (4-hydroxyglucobrassicin, glucobrassicin, 4-methoxyglucobrassicin, and neoglucobrasscin) were detected. Total GSL contents under light conditions 6, 8, and 10 days after sowing (DAS) were 3.2-, 3.9-, and 6.9-fold higher, respectively than those of dark conditions. Interestingly, total GSL contents 2 {85.4 micromol/g dry weight (DW)} to 10 (7.74 micromol/g DW) DAS under dark conditions were gradually decreased. In this study, our results suggest that light affects the levels of GSL in Chinese cabbage seedlings. These results could be useful for obtaining cabbage varieties rich in GSLs.


Bullying among young adolescents: the strong, the weak, and the troubled.
Bullying and being bullied have been recognized as health problems for children because of their association with adjustment problems, including poor mental health and more extreme violent behavior. It is therefore important to understand how bullying and being bullied affect the well-being and adaptive functioning of youth. We sought to use multiple data sources to better understand the psychological and social problems exhibited by bullies, victims, and bully-victims. Analysis of data from a community sample of 1985 mostly Latino and black 6th graders from 11 schools in predominantly low socioeconomic status urban communities (with a 79% response rate). Peer reports of who bullies and who is victimized, self-reports of psychological distress, and peer and teacher reports of a range of adjustment problems. Twenty-two percent of the sample was classified as involved in bullying as perpetrators (7%), victims (9%), or both (6%). Compared with other students, these groups displayed school problems and difficulties getting along with classmates. Despite increased conduct problems, bullies were psychologically strongest and enjoyed high social standing among their classmates. In contrast, victims were emotionally distressed and socially marginalized among their classmates. Bully-victims were the most troubled group, displaying the highest level of conduct, school, and peer relationship problems. To be able to intervene with bullying, it is important to recognize the unique problems of bullies, victims, and bully-victims. In addition to addressing these issues directly with their patients, pediatricians can recommend school-wide antibullying approaches that aim to change peer dynamics that support and maintain bullying.


Effect of aging on urinary concentrating mechanism and vasopressin-dependent cAMP in rats.
The effect of aging on urinary concentrating ability and the pathogenic mechanism involved were investigated in Fischer 344 rats. While the rats had free access to drinking water, 24-mo-old rats were polydipsic and polyuric compared with 6- and 12-mo-old rats. The maximum urinary concentrating ability after 40-58 h of water deprivation was not different between 6- and 12-mo-old rats (Uosmol 2,941 +/- 173 vs. 2,706 +/- 96 (SE) mosmol/kg), but it was significantly decreased in 24-mo-old rats (1,885 +/- 172 mosmol/kg, P less than 0.01). Similarly, although 5 mU/ml vasopressin increased the concentration of cAMP and papillary slices in 12-mo-old rats (delta +2.81 +/- 0.62 pmol/mg tissue, P less than 0.01), the same concentration of vasopressin failed to increase the cAMP concentration in 24-mo-old rats (delta +0.25 +/- 0.21 pmol/mg tissue, P greater than 0.05). In the adenylate cyclase preparation of renal papilla, the response to low concentrations of vasopressin was diminished in 24-mo-old rats. The dose-response curve was shifted to the right and the ED50 concentration of vasopressin was increased in 24-mo-old rats compared with 12-mo-old rats: 1.40 +/- 0.12 mU/ml vasopressin vs. 3.04 +/- 0.22. These results suggest that the decrease in vasopressin-dependent cAMP generation may in part be responsible for the impairment of urinary concentrating ability in 24-mo-old rats.


Inhibition of Acid Sphingomyelinase Allows for Selective Targeting of CD4+ Conventional versus Foxp3+ Regulatory T Cells.
CD4+ Foxp3+ regulatory T cells (Tregs) depend on CD28 signaling for their survival and function, a receptor that has been previously shown to activate the acid sphingomyelinase (Asm)/ceramide system. In this article, we show that the basal and CD28-induced Asm activity is higher in Tregs than in conventional CD4+ T cells (Tconvs) of wild-type (wt) mice. In Asm-deficient (Smpd1-/-; Asm-/-) mice, as compared with wt mice, the frequency of Tregs among CD4+ T cells, turnover of the effector molecule CTLA-4, and their suppressive activity in vitro were increased. The biological significance of these findings was confirmed in our Treg-sensitive mouse model of measles virus (MV) CNS infection, in which we observed more infected neurons and less MV-specific CD8+ T cells in brains of Asm-/- mice compared with wt mice. In addition to genetic deficiency, treatment of wt mice with the Asm inhibitor amitriptyline recapitulated the phenotype of Asm-deficient mice because it also increased the frequency of Tregs among CD4+ T cells. Reduced absolute cell numbers of Tconvs after inhibitor treatment in vivo and extensive in vitro experiments revealed that Tregs are more resistant toward Asm inhibitor-induced cell death than Tconvs. Mechanistically, IL-2 was capable of providing crucial survival signals to the Tregs upon inhibitor treatment in vitro, shifting the Treg/Tconv ratio to the Treg side. Thus, our data indicate that Asm-inhibiting drugs should be further evaluated for the therapy of inflammatory and autoimmune disorders.


[Effect of therapy with blood component (activated autoplasma) on blood glutathione level in patients with vascular abnormalities on the fundus oculi].
Thirty-three patients with central chorioretinal dystrophy, 18 with proliferative diabetic retinopathy, and 15 controls without ocular diseases were examined. All patients were treated by blood components. The treatment started with activation of blood plasma platelets. Glutathione was measured in two portions of plasma: intact and containing platelet activation products. The content of glutathione was higher in activated autoplasma of 21 patients with central chorioretinal dystrophy and 6 patients with diabetic retinopathy than in intact plasma by 107 and 72%, respectively. A decrease in glutathione level in activated autoplasma was observed in 1 patient with central chorioretinal dystrophy and in 9 with diabetic retinopathy. Hence, antioxidant defense is decreased in diabetic retinopathy, since glutathione is involved in reduction of organic hydroperoxides.


Ethnic aged discrimination and disparities in health and social care: a question of social justice.
Older overseas-born Australians of diverse cultural and language backgrounds experience significant disparities in their health and social care needs and support systems. Despite being identified as a 'special needs' group, the ethnic aged in Australia are generally underserved by local health and social care services, experience unequal burdens of disease and encounter cultural and language barriers to accessing appropriate health and social care compared to the average Australian-born population. While a range of causes have been suggested to explain these disparities, rarely has the possibility of cultural racism been considered. In this article, it is suggested that cultural racism be named as a possible cause of ethnic aged disparities and disadvantage in health and social care. It is further suggested that unless cultural racism is named as a structural mechanism by which ethnic aged disparities in health and social care have been created and maintained, redressing them will remain difficult.


Multiple cis-acting sequences contribute to evolved regulatory variation for Drosophila Adh genes.
Drosophila affinidisjuncta and Drosophila hawaiiensis are closely related species that display distinct tissue-specific expression patterns for their homologous alcohol dehydrogenase genes (Adh genes). In Drosophila melanogaster transformants, both genes are expressed at high levels in the larval and adult fat bodies, but the D. affinidisjuncta gene is expressed 10-50-fold more strongly in the larval and adult midguts and Malpighian tubules. The present study reports the mapping of cis-acting sequences contributing to the regulatory differences between these two genes in transformants. Chimeric genes were constructed and introduced into the germ line of D. melanogaster. Stage- and tissue-specific expression patterns were determined by measuring steady-state RNA levels in larvae and adults. Three portions of the promoter region make distinct contributions to the tissue-specific regulatory differences between the native genes. Sequences immediately upstream of the distal promoter have a strong effect in the adult Malpighian tubules, while sequences between the two promoters are relatively important in the larval Malpighian tubules. A third gene segment, immediately upstream of the proximal promoter, influences levels of the proximal Adh transcript in all tissues and developmental stages examined, and largely accounts for the regulatory difference in the larval and adult midguts. However, these as well as other sequences make smaller contributions to various aspects of the tissue-specific regulatory differences. In addition, some chimeric genes display aberrant RNA levels for the whole organism, suggesting close physical association between sequences involved in tissue-specific regulatory differences and those important for Adh expression in the larval and adult fat bodies.


Changes in beta-adrenergic receptor subtypes in Alzheimer-type dementia.
Using ligand binding techniques, we studied beta-adrenergic receptor subtypes in brains obtained at autopsy from seven histologically normal controls and seven histopathologically verified cases with Alzheimer-type dementia (ATD). Inhibition of [3H]dihydroalprenolol [( 3H]DHA) binding by the selective beta 1 antagonist, metoprolol, results in nonlinear Hofstee plots, suggesting the presence of the two receptor subtypes in the human brain. The calculated ratios of beta 1/beta 2-adrenergic receptors in control brains are as follows: frontal cortex, 49:51; temporal cortex, 31:69; hippocampus, 66:34; thalamus, 23:77; putamen, 70:30; caudate, 48:52; nucleus basalis of Meynert (NbM), 43:57; cerebellar hemisphere, 25:75. Compared with the controls, total concentrations of beta-adrenergic receptors were significantly reduced only in the thalamus of the ATD brains. beta 1-Adrenergic receptor concentrations were significantly reduced in the hippocampus and increased in the NbM and cerebellar hemisphere, whereas beta 2-adrenergic receptor concentrations were significantly reduced in the thalamus, NbM, and cerebellar hemisphere and increased in the hippocampus and putamen of the ATD brains. These results suggest that beta 1- and beta 2-adrenergic receptors are present in the human brain and that there are significant changes in both receptor subtypes in selected brain regions in patients with ATD.


Racial/ethnic, socioeconomic, and geographic survival disparities in adolescents and young adults with primary central nervous system tumors.
Disparities in survival by race/ethnicity, socioeconomic status (SES), and geography in adolescent and young adult (AYA) patients with central nervous system (CNS) tumors have not been well studied. A retrospective cohort study utilizing the Surveillance, Epidemiology, and End Results (SEER) database was conducted for AYA patients diagnosed with primary CNS tumors. Adjusted hazard ratios (aHR) were calculated using a multivariate Cox proportional hazard model to evaluate the association between race/ethnicity, SES, rurality, and hazard of death. All minority groups showed an increased hazard of death with greatest disparities in the high-grade glioma cohort. Lower SES was associated with an increased hazard of death in non-Hispanic White (NHW) patients (aHR 1.12; 95% confidence interval [CI] 1.01-1.24), non-Hispanic Black (NHB) patients (aHR 1.34; 95% CI 1.00-1.80), and patients aged 25-29 years (aHR 1.29; 95% CI 1.07-1.55). Mediation analysis showed an indirect effect of SES on the effect of race/ethnicity on the hazard of death only among NHB patients, with SES accounting for 33.7% of the association between NHB and hazard of death. Rurality was associated with an increased hazard of death for patients in the lowest SES tertile (aHR 1.31; 95% CI 1.08-1.59) and NHW patients (aHR 1.20; 95% CI 1.08-1.34). Patients identified as a racial/ethnic minority, patients with a lower SES, and patients residing in rural areas had an increased hazard of death. Further studies are needed to understand and address the biological, psychosocial, societal, and economic factors that impact AYA neuro-oncology patients at highest risk of experiencing poorer outcomes.


Antimicrobial therapy escalation and hospital mortality among patients with health-care-associated pneumonia: a single-center experience.
Patients with health-care-associated pneumonia (HCAP) are frequently infected with a resistant pathogen and receive inappropriate empiric antibiotics (ie, pathogens resistant to administered treatment). Initial inappropriate treatment has been shown to increase hospital mortality. It is not known whether escalation in response to culture results mitigates this risk. We identified patients admitted with a culture-positive pneumonia between January 2003 and December 2005. HCAP patients met one or more of the following criteria indicating ongoing contact with the health-care system: recent hospitalization (< or = 12 months), admission from a nursing home, immunosuppression, or long-term dialysis. We compared survivors to nonsurvivors among those patients with HCAP still hospitalized beyond 48 h. Of 431 HCAP patients, 396 patients (92%) were alive and still hospitalized beyond 48 h. The crude mortality rate was 21.5%. Compared to survivors, nonsurvivors were significantly more likely to be treated with inappropriate empiric antibiotics (37.6% vs 24.1%, p = 0.013). Although mortality was higher among patients receiving inappropriate than appropriate therapy (30.0% vs 18.3%, p = 0.013), this difference was more pronounced among nonbacteremic patients (odds ratio [OR], 2.45; 95% confidence interval [CI], 1.26 to 4.75) than bacteremic patients (OR, 1.25; 95% CI, 0.41 to 3.57). In a logistic regression, inappropriate empiric antibiotic treatment among nonbacteremic patients was independently associated with mortality (OR, 2.88; 95% CI, 1.46 to 5.67); treatment escalation did not attenuate the risk of death. Among HCAP patients alive and hospitalized beyond 48 h, hospital mortality was high and, in the absence of bacteremia, greater with initial inappropriate antibiotic treatment. Despite subsequent escalation, initial inappropriate antibiotic choice nearly tripled the risk of hospital death.


Polycyclic aromatic hydrocarbon coated onto Fe(2)O(3) particles: assessment of cellular membrane damage and antioxidant system disruption in human epithelial lung cells (L132) in culture.
The aim of this study was to investigate the oxidative effects of Fe(2)O(3), benzo(a)pyrene (B(a)P) and pyrene, alone or in association (B(a)P or pyrene coated onto Fe(2)O(3) particles), in normal human embryonic lung epithelial cells (L132) in culture. We evaluated: (i) membrane integrity, through fatty acid release (stearic acid, oleic acid, linoleic and linolenic acids, homolinolenic acid, arachidonic acid) and malondialdehyde (MDA) production; and (ii) antioxidant status, through enzymatic and non-enzymatic antioxidant defenses (superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR), glutathione status, beta-carotene). Fe(2)O(3) did not induce any change in L132 cells. In pyrene-treated cells, SOD induction (P<0. 05), glutathione oxidation (P<0.05) and beta-carotene consumption (P<0.001) may counteract free radicals (FR)-induced damage. However, in B(a)P-incubated cells, SOD inactivation (P<0.05), GR increases (P<0.05), glutathione oxidation (P<0.05) and beta-carotene decreases (P<0.001) showed high disruption of antioxidants, thereby allowing FR-induced damage (i.e. arachidonic acid release, P<0.01; MDA production, P<0.01). Our main finding was that both associations caused higher FR-induced damage (i.e. MDA production, P<0.001; SOD inactivation, P<0.01) than either chemical alone. Several mechanisms could account for this result: enhanced uptake of Fe(2)O(3) particles and/or greater availability of polycyclic aromatic hydrocarbons (PAHs). We hypothesized also that Fe(2)O(3) and polycyclic aromatic hydrocarbons are more deleterious by virtue of their associations being able to produce higher oxidative effects than either chemical alone.


Predictability of spatio-temporal patterns in a lattice of coupled FitzHugh-Nagumo oscillators.
In many biological systems, variability of the components can be expected to outrank statistical fluctuations in the shaping of self-organized patterns. In pioneering work in the late 1990s, it was hypothesized that a drift of cellular parameters (along a 'developmental path'), together with differences in cell properties ('desynchronization' of cells on the developmental path) can establish self-organized spatio-temporal patterns (in their example, spiral waves of cAMP in a colony of Dictyostelium discoideum cells) starting from a homogeneous state. Here, we embed a generic model of an excitable medium, a lattice of diffusively coupled FitzHugh-Nagumo oscillators, into a developmental-path framework. In this minimal model of spiral wave generation, we can now study the predictability of spatio-temporal patterns from cell properties as a function of desynchronization (or 'spread') of cells along the developmental path and the drift speed of cell properties on the path. As a function of drift speed and desynchronization, we observe systematically different routes towards fully established patterns, as well as strikingly different correlations between cell properties and pattern features. We show that the predictability of spatio-temporal patterns from cell properties contains important information on the pattern formation process as well as on the underlying dynamical system.


Drug taking beliefs of Australian adolescents: a pilot study.
In this study adolescents offered their insights and perspectives of factors associated with adolescent illicit drug taking intentions. The factors explored were identified using a cross-disciplinary approach involving the Theory of Planned Behavior (TPB) and criminological theories, and these formed the framework for data analysis. Interviews with 24 students aged 15-17 found that adolescents' beliefs to drug taking attitudes, subjective norms, perceived behavioral control, moral norms, negative affect, and reputation enhancement involved a number a sub-themes that provided an in-depth understanding of the association of these components to intended drug use. The incorporation of these elements in drug education programs could be an effective approach in prevention interventions for adolescent drug use.


Controlled assembly of plasmonic colloidal nanoparticle clusters.
Coupling of localized surface plasmon resonances results in singular effects at the void space between noble metal nanoparticles. However, implementation of practical applications based on plasmon coupling calls for the high yield production of metal nanoparticle clusters (dimers, trimers, tetramers, …) with small gaps. Therefore, controlled assembly using colloid chemistry methods is an emerging and promising field. We present a brief overview over the controlled assembly of plasmonic nanoparticle clusters by colloid chemistry methods, together with a description of their plasmonic properties and some applications, with an emphasis in sensing through surface-enhanced Raman scattering spectroscopy for bio-detection purposes. We point out the important role of separation methods to obtain colloidal clusters in high yield. A special encouragement to explore assembly of anisotropic building blocks is pursued.


Usefulness of Stereotactic Body Radiation Therapy for Treatment of Adrenal Gland Metastases.
Purpose: This study aimed to describe our institutional experience in the use of stereotactic body radiation therapy (SBRT) for the management of adrenal gland metastases from multiple primary cancers. Materials and Methods: We retrospectively reviewed 31 patients who underwent SBRT as treatment for 33 adrenal gland lesions in the academic radiotherapy department of Oscar Lambret cancer center between May 2011 and September 2018. The primary study endpoints were 1- and 2-year local control rates, defined as the absence of progression at the treatment site based on the response evaluation criteria in solid tumors (RECIST). Toxicities were graded in accordance with the Common Terminology Criteria for Adverse Events version 4.03. Results: The average tumor volume was 33.5 cm3 (standard deviation: 51.7 cm3), and the prescribed dose ranged from 30 to 55 Gy given in 3-9 fractions. The median biological effective dose was 112.5 Gy (range: 45-115.5 Gy), assuming α/β = 10. Considering progression at distant sites or death as competing events, the 1- and 2-year actuarial local control rates were 96.5% (95% confidence interval: 84.9-99.7) and 92.6% (95% confidence interval: 79.2-98.7), respectively. According to RECIST, a complete response was achieved in 10 (32.3%) lesions, a partial response in 10 (32.3%) lesions, and stability in 8 (25.8%) lesions. Three patients presented with local relapse at 8.8, 14, and 49.4 months. After a median follow-up of 18 months (range: 4.4-66.4), the median overall survival was 33.5 months (95% confidence interval: 17-not reached), while the median progression-free survival was 7.4 months (95% confidence interval: 3.8-14.1). Treatment-related toxicity was grade 1 or 2 in 42.4% of patients, including nausea (27.3%), abdominal pain (18.2%), vomiting (15.2%), and asthenia (9.1%). None of the patients developed acute grade ≥3 or late toxicity. Conclusion: SBRT seems to be a safe and effective treatment for adrenal gland metastases in patients whose primary tumor and metastatic spread are controlled by systemic treatment. With a 2-year local control rate of 92.6%, SBRT may be considered as one of the first-line treatments in oligometastatic patients with adrenal metastases.


Study Protocol: Transitions in Adolescent Girls (TAG).
Despite recent studies linking pubertal processes to brain development, as well as research demonstrating the importance of both pubertal and neurodevelopmental processes for adolescent mental health, there is limited knowledge of the full pathways and mechanisms behind the emergence of mental illnesses such as depression and anxiety disorders in adolescence. The Transitions in Adolescent Girls (TAG) study aims to understand the complex relationships between pubertal development, brain structure and connectivity, the behavioral and neural correlates of social and self-perception processes, and adolescent mental health in female adolescents. The TAG study includes 174 female adolescents aged 10.0 to 13.0 years, recruited from the local community in Lane County, Oregon, USA. The participants, along with a parent/guardian, will complete three waves of assessment over the course of 3 years; the third wave is currently underway. Each wave includes collection of four saliva samples (one per week) and one hair sample for the assessment of hormone levels and immune factors; an MRI session including structural, diffusion, resting-state functional and task-based functional scans; the Kiddie Schedule for Affective Disorders and Schizophrenia (K-SADS), a diagnostic interview on current and lifetime mental health; production of a short self-narrative video; and measurement of height, weight, and waist circumference. The functional MRI tasks include a self-evaluation paradigm and a self-disclosure paradigm. In addition, adolescents and their parents/guardians complete a number of surveys to report on the adolescent's pubertal development, mental health, social environment and life events; adolescents also report on various indices of self-perception and social-emotional functioning. The knowledge gained from this study will include developmental trajectories of pubertal, neurological, and social processes and their roles as mechanisms in predicting emergence of mental illness in female adolescents. This knowledge will help identify modifiable, developmentally specific risk factors as targets for early intervention and prevention efforts.


[Mental and physical development of children living in the area of monitoring pollution from a plant for processing and long-term storage of radioactive waste].
Environmental conditions in the area hosting a plant for processing and long storage of low- to moderately-active radioactive waste are described as reasonably safe. Residence in the area does not exert negative influence on the physical and mental development of children. Several indicators of physical development give better estimates than at the control territory. The difference can be accounted for by a better social situation in the study area (housing conditions, financial standing, food patterns, general lifestyle).


Integrated Analysis Of Immunotherapy Treated Clear Cell Renal Cell Carcinomas: An Exploratory Study.
Molecular or immunological differences between responders and nonresponders to immune checkpoint inhibitors (ICIs) of clear cell renal cell carcinomas (ccRCCs) remain incompletely understood. To address this question, we performed next-generation sequencing, methylation analysis, genome wide copy number analysis, targeted RNA sequencing and T-cell receptor sequencing, and we studied frequencies of tumor-infiltrating CD8+ T cells, presence of tertiary lymphoid structures (TLS) and PD-L1 expression in 8 treatment-naive ccRCC patients subsequently treated with ICI (3 responders, 5 nonresponders). Unexpectedly, we identified decreased frequencies of CD8+ tumor-infiltrating T cells and TLS, and a decreased expression of PD-L1 in ICI responders when compared with nonresponders. However, neither tumor-specific genetic alterations nor gene expression profiles correlated with response to ICI or the observed immune features. Our results underline the challenge to stratify ccRCC patients for immunotherapy based on routinely available pathologic primary tumor material, even with advanced technologies. Our findings emphasize the analysis of pretreated metastatic tissue in line with recent observations describing treatment effects on the tumor microenvironment. In addition, our data call for further investigation of additional parameters in a larger ccRCC cohort to understand the mechanistic implications of the observed differences in tumor-infiltrating CD8+ T cells, TLS, and PD-L1 expression.


Immortalization of inbred rabbit keratinocytes from a Shope papilloma and tumorigenic transformation of the cells by EJ-ras.
An immortalized cell line of keratinocytes, named SPG1-3, was established from a papilloma induced from cottontail rabbit papillomavirus (CRPV)-infected inbred rabbit skin. The cells have reached 60 passages in culture and are still growing well, but they are not tumorigenic in athymic mice. Although CRPV DNA was present as extrachromosomal episomes in the papilloma from which the cell line was derived from a single colony of keratinocytes, there was no CRPV DNA detectable in the cells. Three sub-cell lines of SPG1-3EJ, SPG1-3EJ1 and SPG1-3EJ2 were then established from the EJ-ras transfected SPG1-3 cells. All of the three sub-lines contained both EJ-ras DNA and a 1.2 kb transcript of EJ-ras, and they are malignantly tumorigenic in athymic mice. These data indicate that CRPV genome and its expression might be essential for the initiation and maintenance of neoplasia, but not for the maintenance of immortalization of the tumor-derived cells. In addition, some oncogenes such as EJ-ras may play an essential role in tumorigenic and malignant conversion of the immortalized cells. These cell lines derived from inbred rabbit skin may provide a useful in vitro system for better understanding of the oncogenic processes of papillomavirus-involved neoplastic progression by transfecting the cells with CRPV genes and serial transplantation to the inbred rabbits for studying host immune responses to the viral oncogenic potential.


Application of polyaniline/sol-gel derived tetraethylorthosilicate films to an amperometric lactate biosensor.
The electrochemical entrapment of polyaniline (PANI) onto sol-gel derived tetraethylorthosilicate (TEOS) films deposited onto indium-tin-oxide (ITO) coated glass has been utilized for immobilization of lactate dehydrogenase (LDH). The performance of these sol-gel/PANI/LDH electrodes has been investigated as a function of the lactate concentration, applied potential, pH of the medium and interferents. The amperometric response of the electrodes under optimum conditions exhibited a linear relationship from 1 mM to 4 mM. An attempt has been made to extend the linearity up to 10 mM for lactate by coating an external layer of polyvinyl chloride (PVC) over the sol-gel/PANI/LDH electrodes with a correlation coefficient of 0.89. These sol-gel/PANI/LDH electrodes have a response time of about 60 s, a shelf life of about 8 weeks at 0-4 degrees C and have implications in a lactate biosensor.


Dopamine in hypophysial portal plasma and prolactin in systemic plasma of rats treated with 5-hydroxytryptamine.
Intracerebroventricularly administered 5-hydroxytryptamine (5HT) altered the release of dopamine into pituitary stalk blood and of PRL into the systemic circulation of male rats. The concentration of dopamine in pituitary stalk plasma of rats given 0.5 or 5.0 microgram 5HT was 0.32 +/- 0.06 (mean +/- SE) or 0.18 +/- 0.04 ng/ml, respectively, and was significantly less than that in vehicle-treated animals (0.80 +/- 0.04 ng/ml). Relative to the mean level of PRL in animals injected with the solvent vehicle, the mean concentration of PRL in central venous plasma of rats given 0.5 or 5.0 microgram 5HT was increased 4-fold or 13-fold, respectively. Other rats were infused iv with dopamine for 45 min. After 15 min of infusion, the mean concentration of dopamine in arterial plasma was several times that of dopamine in pituitary stalk plasma of rats not infused with dopamine. Even so, this high concentration of dopamine did not prevent the 5HT-induced release of PRL. We conclude that 5HT modulates the secretion of PRL through a stimulatory mechanism that is effective in the presence of a high plasma concentration of dopamine, a PRL-release inhibiting factor, and suggest that 5HT causes the release of a hypothalamic substance(s) that stimulates release of PRL, i.e. a PRL-releasing factor.


Current status of radiotherapy with proton and light ion beams.
Several model studies have shown potential clinical advantages with charged particles (protons and light ions) compared with 3D-conformal radiotherapy (3D-CRT) and intensity-modulated radiotherapy (IMRT) in many disease sites. The newly developed intensity-modulated proton therapy (IMPT) often yields superior dose distributions to photon IMRT, with the added advantage of a significant reduction in the volume of healthy normal tissues exposed to low-to-medium doses. Initially, the major emphasis in clinical research for proton and light ion therapy was dose escalation for inherently radioresistant tumors, or for lesions adjacent to critical normal structures that constrained the dose that could be safely delivered with conventional x-ray therapy. Since the advent of IMRT the interest in particle therapy has gradually shifted toward protocols aimed at morbidity reduction. Lately the emphasis has mostly been placed on the potential for reduced risk of radiation-induced carcinogenesis with protons. Compared with 3D-CRT, a 2-fold increase has been theoretically estimated with the use of IMRT due to the larger integral volumes. In the pediatric setting, due to a higher inherent susceptibility of tissues, the risk could be significant, and the benefits of protons have been strongly emphasized in the literature. There is a significant expansion of particle therapy facilities around the world. Increasing public awareness of the potential benefits of particle therapy and wider accessibility for patients require that treating physicians stay abreast of the clinical indications of this radiotherapy modality. The article reviews the available literature for various disease sites in which particle therapy has traditionally been considered to offer clinical advantages and to highlight current lines of clinical research. The issue of radiation-induced second malignancies is examined in the light of the controversial epidemiological evidence available. The cost-effectiveness of particle therapy is also discussed.


Association of Nrf2 polymorphism haplotypes with acute lung injury phenotypes in inbred strains of mice.
Nrf2 is a master transcription factor for antioxidant response element (ARE)-mediated cytoprotective gene induction. A protective role for pulmonary Nrf2 was determined in model oxidative disorders, including hyperoxia-induced acute lung injury (ALI). To obtain additional insights into the function and genetic regulation of Nrf2, we assessed functional single nucleotide polymorphisms (SNPs) of Nrf2 in inbred mouse strains and tested whether sequence variation is associated with hyperoxia susceptibility. Nrf2 SNPs were compiled from publicly available databases and by re-sequencing DNA from inbred strains. Hierarchical clustering of Nrf2 SNPs categorized the strains into three major haplotypes. Hyperoxia susceptibility was greater in haplotypes 2 and 3 strains than in haplotype 1 strains. A promoter SNP -103 T/C adding an Sp1 binding site in haplotype 2 diminished promoter activation basally and under hyperoxia. Haplotype 3 mice bearing nonsynonymous coding SNPs located in (1862 A/T, His543Gln) and adjacent to (1417 T/C, Thr395Ile) the Neh1 domain showed suppressed nuclear transactivation of pulmonary Nrf2 relative to other strains, and overexpression of haplotype 3 Nrf2 showed lower ARE responsiveness than overexpression of haplotype 1 Nrf2 in airway cells. Importantly, we found a significant correlation of Nrf2 haplotypes and hyperoxic lung injury phenotypes. The results indicate significant influence of Nrf2 polymorphisms and haplotypes on gene function and hyperoxia susceptibility. Our findings further support Nrf2 as a genetic determinant in ALI pathogenesis and provide useful tools for investigators who use mouse strains classified by Nrf2 haplotypes to elucidate the role for Nrf2 in oxidative disorders.


Bifunctional MIL-53(Fe) with pyrophosphate-mediated peroxidase-like activity and oxidation-stimulated fluorescence switching for alkaline phosphatase detection.
Alkaline phosphatase (ALP) is extensively used as a clinical biomarker because of its close relevance with a variety of diseases. Thus, exploring reliable and practical methods for ALP analysis is of great significance. In the present work, we explored MIL-53(Fe) as a bifunctional platform with pyrophosphate (PPi)-mediated peroxidase-like activity and oxidation-stimulated fluorescence switching for ALP sensing. The proposed MIL-53(Fe) could exhibit favorable peroxidase-mimicking activity to catalytically decompose H2O2 to hydroxyl radicals, which had strong oxidizing ability to oxidize the terephthalic acid bridging ligand, resulting in the oxidation-stimulated turn-on fluorescence of MIL-53(Fe) itself. Due to the strong coordination interaction between PPi and Fe3+, the former with a relatively large molecular structure was able to inhibit the catalytic activity of MIL-53(Fe) via capping active Fe3+ sites, leading to the suppression of its self-fluorescence response. When ALP was present, it could hydrolyze the PPi inhibitor and restore the dual functions of MIL-53(Fe) to provide fluorescence again. With the above principle, highly sensitive and selective determination of ALP with a linear scope of 2-80 U L-1 and a detection limit down to 0.7 U L-1 was achieved. The MIL-53(Fe) was also demonstrated to be very reliable in measuring the target in human serum, indicating its great promise as an integrated tool for ALP detection in clinical practice.


[Cytostatic polychemotherapy in the aged].
Adherence to the dose or the need for dose reduction and the duration of treatment intervals were determined retrospectively for 1446 chemotherapy courses in 291 patients with malignant lymphoma and breast cancer. In patients over 60 years of age treated with the COP and COPP regimes there was a significantly higher frequency of deviation from the standard regime than in younger patients; a similar situation was seen in patients with breast cancer. The cause in the elderly patients was presumably due to the higher incidence of non-oncological diseases. In a group treated using the CHOP-scheme there was no difference in comparison with the reference group. In this group patients with severe pre-existent diseases were excluded before treatment. The results indicate that age itself is not a major risk factor for a combination chemotherapy. Pre-existent diseases play a substantial role in the toxicity of cytostatics.


A new species of Henneguya (Myxozoa) in the big-eyed scad (Selar crumenophthalmus) from Hawaii.
We describe a new myxozoan, Henneguya akule n. sp., infecting the carangid fish Selar crumenophthalmus in Hawaii. Spores were found only in the aortic bulb, characterized by elliptical capsule with 2 tails, and pyriform polar capsules that angled toward the anterior end of the spore. Polar filaments had 3-4 coils. Parasites were present in apparently healthy fishes and caused no evident gross pathology. On microscopy, parasites evinced a mild inflammatory response in the host characterized by accumulations of eosinophilic fibrillar material around spores and a mononuclear infiltrate in the adventitia of the bulbus arteriosus. Overall prevalence was 20%, and prevalence between 2001 and 2006 ranged from 12 to 27%, but did not differ significantly between years. In contrast, prevalence of infection was highest in south-central Oahu. There was no relationship between infection status and body condition or gender of fish, and infection was absent in the smallest and largest fishes. Phylogenetically, H. akule n. sp. is most closely related to other Henneguya species infecting the heart of marine fishes based on ribosomal DNA analysis. This is the first documentation of a myxozoan parasite in marine fishes from Hawaii.


NF-kappaB-inducing kinase is involved in the activation of the CD28 responsive element through phosphorylation of c-Rel and regulation of its transactivating activity.
Previous evidence suggested that NF-kappaB-inducing kinase (NIK) might regulate IL-2 synthesis. However, the molecular mechanism is not understood. In this study, we show that NIK is involved in CD3 plus CD28 activation of IL-2 transcription. Splenic T cells from aly/aly mice (that have a defective NIK protein) have a severe impairment in IL-2 and GM-CSF but not TNF secretion in response to CD3/CD28. This effect takes place at the transcriptional level as overexpression of alyNIK inhibits IL-2 promoter transcription. NIK activates the CD28 responsive element (CD28RE) of the IL-2 promoter and strongly synergizes with c-Rel in this activity. We found that NIK interacts with the N-terminal domain of c-Rel, mapping this interaction to aa 771-947 of NIK. Moreover, NIK phosphorylates the c-Rel C-terminal transactivation domain (TAD) and induces Gal4-c-Rel-transactivating activity. Anti-CD28 activated Gal4-c-Rel transactivation activity, and this effect was inhibited by a NIK-defective mutant. Deletion studies mapped the region of c-Rel responsive to NIK in aa 456-540. Mutation of several serines, including Ser471, in the TAD of c-Rel abrogated the NIK-enhancing activity of its transactivating activity. Interestingly, a Jurkat mutant cell line that expresses one of the mutations of c-Rel (Ser471Asn) has a severe defect in IL-2 and CD28RE-dependent transcription in response to CD3/CD28 or to NIK. Our results support that NIK may be controlling CD28RE-dependent transcription and T cell activation by modulating c-Rel phosphorylation of the TAD. This leads to more efficient transactivation of genes which are dependent on CD28RE sites where c-Rel binds such as the IL-2 promoter.


CT assessment of cerebral perfusion: experimental validation and initial clinical experience.
To validate a dynamic single-section computed tomographic (CT) method to measure cerebral blood volume (CBV) and cerebral blood flow (CBF) by using a noncarotid artery as the input and to demonstrate the feasibility of this method in a pilot series of patients. Twelve dynamic contrast material-enhanced CT studies were performed in beagles. CBV, CBF, and mean transit time (MTT) values were calculated by using an internal carotid artery (ICA) and a noncarotid artery as the input artery to the brain. Patient studies with use of the radial artery as the input were performed (a) repetitively in two patients after subarachnoid hemorrhage, (b) in a patient with a symptomatic ICA occlusion before and after the intravenous injection of 1 g of acetazolamide, and (c) in a patient with a malignant brain tumor. Linear regression analyses revealed highly significant correlations (P < .001) between CBV (r, 0.98; slope, 0.96), CBF (r, 0.89; slope, 0.87), and MTT (r, 0.80; slope, 0.76) values calculated with the ICA and the noncarotid inputs. The CT-derived patient data correlated well with ancillary clinical and neuroradiologic findings. Dynamic single-section CT scanning to measure CBV and CBF on the basis of a noncarotid input is a highly accessible and cost-effective blood flow measurement technique.


RPE-based zone of normality for use in evaluating graded exercise test responses in women.
The purposes of this study were to identify a zone of normality based upon ratings of perceived exertion (RPE) and compare the RPE of women with coronary artery disease to this zone. 45 healthy women (34.8 +/- 9.1 yr.) completed a Bruce treadmill test. RPE were estimated during the last minute of each workload. The zone of normality was established as the 95% confidence interval spanning the average line when RPE was regressed against MET. 51 women (66.6 +/- 9.3 yr.) with coronary artery disease completed a symptom-limited treadmill test. 31 of the women with coronary artery disease were taking beta blockers. RPE were compared to the RPE/MET relation of the zone. 57% of the women with coronary artery disease estimated RPE above the zone during the treadmill test. RPE responses above the zone of normality indicate an elevated perceptual strain for a given energy expenditure (i.e., MET). The zone of normality might be used as an aid in discriminating between normal and abnormal RPE responses during graded treadmill tests by women with coronary artery disease.


α-Synuclein emulsifies TDP-43 prion-like domain-RNA liquid droplets to promote heterotypic amyloid fibrils.
Many neurodegenerative diseases including frontotemporal lobar degeneration (FTLD), Lewy body disease (LBD), multiple system atrophy (MSA), etc., show colocalized deposits of TDP-43 and α-synuclein (αS) aggregates. To understand whether these colocalizations are driven by specific molecular interactions between the two proteins, we previously showed that the prion-like C-terminal domain of TDP-43 (TDP-43PrLD) and αS synergistically interact to form neurotoxic heterotypic amyloids in homogeneous buffer conditions. However, it remains unclear if αS can modulate TDP-43 present within liquid droplets and biomolecular condensates called stress granules (SGs). Here, using cell culture and in vitro TDP-43PrLD - RNA liquid droplets as models along with microscopy, nanoscale AFM-IR spectroscopy, and biophysical analyses, we uncover the interactions of αS with phase-separated droplets. We learn that αS acts as a Pickering agent by forming clusters on the surface of TDP-43PrLD - RNA droplets. The aggregates of αS on these clusters emulsify the droplets by nucleating the formation of heterotypic TDP-43PrLD amyloid fibrils, structures of which are distinct from those derived from homogenous solutions. Together, these results reveal an intriguing property of αS to act as a Pickering agent while interacting with SGs and unmask the hitherto unknown role of αS in modulating TDP-43 proteinopathies.


Generalised granulomatous disease in a horse.
A 6-year-old gelding was referred with a 3-month history of recurrent fever, inappetance, lethargy and weight loss. On clinical examination major findings were depression, thin condition, thrombophlebitis, nodules on the scrotal skin, leukocytosis, hyperfibrinogenaemia and hyperglobulinaemia. Pleural fluid and areas of lung consolidation were seen on ultrasonographic examination of the thorax. A diagnosis of chronic respiratory disease was made. Initially there was a response to antibiotic therapy but the horse was presented 3 months later with continued weight loss, recurrent fever and multifocal skin lesions, characterised by scales, crusts and nodules, affecting the nasal bridge, jugular grooves, ventral neck, withers, scrotum, prepuce, and medial gaskins. Histological evaluation of skin biopsies indicated a granulomatous reaction. On ultrasonographic examination of the thorax multiple hypoechoic lesions consistent with granulomas were seen in both lungs. A diagnosis of generalised granulomatous disease was made. The horse was euthanased at the owner's request. On necropsy examination the main findings were multiple nodules of fibrotic granulomatous inflammation in the lung, heart, liver, gastrointestinal tract and mesenteric lymph nodes, supporting the diagnosis of generalised granulomatous disease.


Practice Current: When do you suspect autoimmune encephalitis and what is the role of antibody testing?
Diagnosing autoimmune encephalitis (AE) is complicated by several factors, including issues with availability, sensitivity, and specificity of antibody testing, particularly with variability in assay techniques and new antibodies being rapidly identified; nonspecific findings on MRI, EEG, and lumbar puncture; and competing differential diagnoses. Through case-based discussions with 3 experts from 3 continents, this article discusses the challenges of AE diagnosis, important clinical characteristics of AE, preferences for methods of autoantibody testing and interpretation, and treatment-related questions. In particular, we explore the following question: If a patient's clinical presentation seems consistent with AE but antibody testing is negative, can one still diagnose the patient with AE? Furthermore, what factors does one consider when making this determination, and should treatment proceed independent of antibody testing in suspected cases? The same case-based questions were posed to the rest of our readership in an online survey, the results of which are also presented.


Cannabinoid CB2 receptor as a new phototherapy target for the inhibition of tumor growth.
The success of targeted cancer therapy largely relies upon the selection of target and the development of efficient therapeutic agents that specifically bind to the target. In the current study, we chose a cannabinoid CB2 receptor (CB2R) as a new target and used a CB2R-targeted photosensitizer, IR700DX-mbc94, for phototherapy treatment. IR700DX-mbc94 was prepared by conjugating a photosensitizer, IR700DX, to mbc94, whose binding specificity to CB2R has been previously demonstrated. We found that phototherapy treatment using IR700DX-mbc94 greatly inhibited the growth of CB2R positive tumors but not CB2R negative tumors. In addition, phototherapy treatment with nontargeted IR700DX did not show significant therapeutic effect. Similarly, treatment with IR700DX-mbc94 without light irradiation or light irradiation without the photosensitizer showed no tumor-inhibitory effect. Taken together, IR700DX-mbc94 is a promising phototherapy agent with high target-specificity. Moreover, CB2R appears to have great potential as a phototherapeutic target for cancer treatment.


EPR and Mössbauer spectroscopy of intact mitochondria isolated from Yah1p-depleted Saccharomyces cerevisiae.
Yah1p, an [Fe 2S 2]-containing ferredoxin located in the matrix of Saccharomyces cerevisiae mitochondria, functions in the synthesis of Fe/S clusters and heme a prosthetic groups. EPR, Mossbauer spectroscopy, and electron microscopy were used to characterize the Fe that accumulates in Yah1p-depleted isolated intact mitochondria. Gal- YAH1 cells were grown in standard rich media (YPD and YPGal) under O 2 or argon atmospheres. Mitochondria were isolated anaerobically, then prepared in the as-isolated redox state, the dithionite-treated state, and the O 2-treated state. The absence of strong EPR signals from Fe/S clusters when Yah1p was depleted confirms that Yah1p is required in Fe/S cluster assembly. Yah1p-depleted mitochondria, grown with O 2 bubbling through the media, accumulated excess Fe (up to 10 mM) that was present as 2-4 nm diameter ferric nanoparticles, similar to those observed in mitochondria from yfh1Delta cells. These particles yielded a broad isotropic EPR signal centered around g = 2, characteristic of superparamagnetic relaxation. Treatment with dithionite caused Fe (3+) ions of the nanoparticles to become reduced and largely exported from the mitochondria. Fe did not accumulate in mitochondria isolated from cells grown under Ar; a significant portion of the Fe in these organelles was in the high-spin Fe (2+) state. This suggests that the O 2 used during growth of Gal- YAH1 cells is responsible, either directly or indirectly, for Fe accumulation and for oxidizing Fe (2+) --> Fe (3+) prior to aggregation. Models are proposed in which the accumulation of ferric nanoparticles is caused either by the absence of a ligand that prevents such precipitation in wild-type mitochondria or by a more oxidizing environment within the mitochondria of Yah1p-depleted cells exposed to O 2. The efficacy of reducing accumulated Fe along with chelating it should be considered as a strategy for its removal in diseases involving such accumulations.


Effects of ethanol on CRF release in vitro.
Acute ethanol exposure produces activation of the brain-pituitary-adrenal (BPA) axis, resulting in the release of ACTH, beta-endorphin, and glucocorticoids. While elevated levels of plasma glucocorticoids are also found after chronic ethanol administration, plasma ACTH and beta-endorphin are normal or reduced. It is also unclear whether chronic ethanol exposure results in tolerance to the stimulatory effect of ethanol on BPA activity. To determine the site and mechanism of ethanol action on the BPA axis we studied the CRF secretory profile in a superfused rat hypothalamic preparation after chronic ethanol administration in vivo and the CRF responses after acute ethanol exposure in vitro. Superfused hypothalami from normal and pair-fed control rats released CRF-like immunoreactive material (CRF-LI) in a pulsatile manner, with a mean (+/- SE) frequency of 5.1 +/- 0.7 pulses/h. In contrast, the pulse frequency of CRF-LI release from hypothalami of rats receiving chronic ethanol treatment (fed an alcohol-containing liquid diet for 2 weeks) increased dramatically; the basal mean CRF level, pulse amplitude, and pulse duration remained unchanged. Hypothalamic CRF content was decreased. This chronic ethanol exposure also altered the dose-response characteristics of CRF release when ethanol was introduced acutely, as a pulse, into the in vitro preparation. Acute exposure to 20 mg/100 ml ethanol produced greater release of CRF-LI from control hypothalami than from chronic ethanol-exposed hypothalami. A further elevation above basal levels was produced by 200 mg/100 ml ethanol in control, but not ethanol-exposed, hypothalami. Secretion of CRF from ethanol-exposed hypothalami in response to depolarizing concentrations of potassium chloride was suppressed. Chronic ethanol treatment had no effect on CRF-LI and CRF bioactivity responses to stimulation with acetylcholine. These findings suggest the presence of a high frequency pulse-generating mechanism for CRF release in the hypothalamus. This pulsatile secretory mechanism is altered by chronic ethanol exposure of the animals in vivo. Chronic intoxication resulted in tolerance to the stimulatory effect of ethanol on CRF release in vitro.


Agent-based model demonstrates the impact of nonlinear, complex interactions between cytokines on muscle regeneration.
Muscle regeneration is a complex process due to dynamic and multiscale biochemical and cellular interactions, making it difficult to determine optimal treatments for muscle injury using experimental approaches alone. To understand the degree to which individual cellular behaviors impact endogenous mechanisms of muscle recovery, we developed an agent-based model (ABM) using the Cellular Potts framework to simulate the dynamic microenvironment of a cross-section of murine skeletal muscle tissue. We referenced more than 100 published studies to define over 100 parameters and rules that dictate the behavior of muscle fibers, satellite stem cells (SSC), fibroblasts, neutrophils, macrophages, microvessels, and lymphatic vessels, as well as their interactions with each other and the microenvironment. We utilized parameter density estimation to calibrate the model to temporal biological datasets describing cross-sectional area (CSA) recovery, SSC, and fibroblast cell counts at multiple time points following injury. The calibrated model was validated by comparison of other model outputs (macrophage, neutrophil, and capillaries counts) to experimental observations. Predictions for eight model perturbations that varied cell or cytokine input conditions were compared to published experimental studies to validate model predictive capabilities. We used Latin hypercube sampling and partial rank correlation coefficient to identify in silico perturbations of cytokine diffusion coefficients and decay rates to enhance CSA recovery. This analysis suggests that combined alterations of specific cytokine decay and diffusion parameters result in greater fibroblast and SSC proliferation compared to individual perturbations with a 13% increase in CSA recovery compared to unaltered regeneration at 28 days. These results enable guided development of therapeutic strategies that similarly alter muscle physiology (i.e. converting ECM-bound cytokines into freely diffusible forms as studied in cancer therapeutics or delivery of exogenous cytokines) during regeneration to enhance muscle recovery after injury.


Implementing syndromic surveillance: a practical guide informed by the early experience.
Syndromic surveillance refers to methods relying on detection of individual and population health indicators that are discernible before confirmed diagnoses are made. In particular, prior to the laboratory confirmation of an infectious disease, ill persons may exhibit behavioral patterns, symptoms, signs, or laboratory findings that can be tracked through a variety of data sources. Syndromic surveillance systems are being developed locally, regionally, and nationally. The efforts have been largely directed at facilitating the early detection of a covert bioterrorist attack, but the technology may also be useful for general public health, clinical medicine, quality improvement, patient safety, and research. This report, authored by developers and methodologists involved in the design and deployment of the first wave of syndromic surveillance systems, is intended to serve as a guide for informaticians, public health managers, and practitioners who are currently planning deployment of such systems in their regions.


Outcomes of Sentinel Node Biopsy for Women with Breast Cancer After Neoadjuvant Therapy: Systematic Review and Meta-Analysis of Real-World Data.
Evidence on the accuracy of sentinel lymph node biopsy (SLNB) after neoadjuvant therapy (NAT) for patients with breast cancer is inconclusive. This study reviewed the real-world data to determine the acceptability of SLNB after NAT. The study searched for articles in the PubMed, EMBASE, and Cochrane Library databases. The primary outcomes were the identification rate for sentinel lymph nodes (SLNs) and the false-negative rate (FNR) for SLNB. The study also evaluated the FNR in subgroups defined by tumor stage, nodal stage, hormone receptor status, human epidermal growth factor receptor-2 status, tumor response, mapping technique, and number of SLNs removed. The study retrieved 61 prospective and 18 retrospective studies with 10,680 initially cN± patients. The pooled estimate of the identification rate was 0.906 (95 % confidence interval [CI], 0.891-0.922), and the pooled FNR was 0.118 (95 % CI, 0.103-0.133). In subgroup analysis, the FNR was significantly higher for the patients with estrogen receptor (ER)-negative status and fewer than three SLNs removed. The FNR did not differ significantly between the patients with and those without complete tumor response. Among the patients with initial clinical negative axillary lymph nodes, the incidence of node metastasis was 26.8 % (275/1041) after NAT. Real-world evidence indicates that the FNR of SLNB after NAT in breast cancer is 11.8 %, exceeding only slightly the commonly adopted threshold of 10 %. The FNR is significantly higher for patients with ER-negative status and removal of fewer than three SLNs. Using a dual tracer and removing at least three SLNs may increase the accuracy of SLNB after NAT.


Quantifying cellular forces and biomechanical properties by correlative micropillar traction force and Brillouin microscopy.
Cells sense and respond to external physical forces and substrate rigidity by regulating their cell shape, internal cytoskeletal tension, and stiffness. Here we show that the combination of micropillar traction force and noncontact Brillouin microscopy provides access to cell-generated forces and intracellular mechanical properties at optical resolution. Actin-rich cytoplasmic domains of 3T3 fibroblasts showed significantly higher Brillouin shifts, indicating a potential increase in stiffness when adhering on fibronectin-coated glass compared to soft PDMS micropillars. Our findings demonstrate the complementarity of micropillar traction force and Brillouin microscopy to better understand the relation between cell force generation and the intracellular mechanical properties.


A Socially Assistive Robot for Stroke Patients: Acceptance, Needs, and Concerns of Patients and Informal Caregivers.
Stroke patients often contend with long-term physical challenges that require treatment and support from both formal and informal caregivers. Socially Assistive Robots (SARs) can assist patients in their physical rehabilitation process and relieve some of the burden on the informal caregivers, such as spouses and family members. We collected and analyzed information from 23 participants (11 stroke patients and 12 informal caregivers) who participated in a total of six focus-group discussions. The participants responded to questions regarding using a SAR to promote physical exercises during the rehabilitation process: (a) the advantages and disadvantages of doing so; (b) specific needs that they wish a SAR would address; (c) patient-specific adaptations they would propose to include; and (d) concerns they had regarding the use of such technology in stroke rehabilitation. We found that the majority of the participants in both groups were interested in experiencing the use of a SAR for rehabilitation, in the clinic and at home. Both groups noted the advantage of having the constant presence of a motivating entity with whom they can practice their rehabilitative exercises. The patients noted how such a device can assist formal caregivers in managing their workload, while the informal caregivers indicated that such a system could ease their own workload and sense of burden. The main disadvantages that participants noted related to the robot not possessing human abilities, such as the ability to hold a conversation, to physically guide the patient's movements, and to express or understand emotions. We anticipate that the data collected in this study-input from the patients and their family members, including the similarities and differences between their points of view-will aid in improving the development of SARs for rehabilitation, so that they can better suit people who have had a stroke, and meet their individual needs.


Large-scale exploration and analysis of drug combinations.
Drug combinations are a promising strategy for combating complex diseases by improving the efficacy and reducing corresponding side effects. Currently, a widely studied problem in pharmacology is to predict effective drug combinations, either through empirically screening in clinic or pure experimental trials. However, the large-scale prediction of drug combination by a systems method is rarely considered. We report a systems pharmacology framework to predict drug combinations (PreDCs) on a computational model, termed probability ensemble approach (PEA), for analysis of both the efficacy and adverse effects of drug combinations. First, a Bayesian network integrating with a similarity algorithm is developed to model the combinations from drug molecular and pharmacological phenotypes, and the predictions are then assessed with both clinical efficacy and adverse effects. It is illustrated that PEA can predict the combination efficacy of drugs spanning different therapeutic classes with high specificity and sensitivity (AUC = 0.90), which was further validated by independent data or new experimental assays. PEA also evaluates the adverse effects (AUC = 0.95) quantitatively and detects the therapeutic indications for drug combinations. Finally, the PreDC database includes 1571 known and 3269 predicted optimal combinations as well as their potential side effects and therapeutic indications. The PreDC database is available at http://sm.nwsuaf.edu.cn/lsp/predc.php.


Impact of high-normal blood pressure on the risk of cardiovascular disease in a Japanese urban cohort: the Suita study.
Few prospective studies have examined the association between high-normal blood pressure and cardiovascular disease (CVD) in Asia. We examined the impact of high-normal blood pressure on the incidence of CVD in a general urban population cohort in Japan. We studied 5494 Japanese individuals (ages 30 to 79 years without CVD at baseline) after completing a baseline survey who received follow-up through December 2005. Blood pressure categories were defined on the basis of the ESH-ESC 2007 criteria. In 64 391 person-years of follow-up, we documented the incidence of 346 CVD events. The frequencies of high-normal blood pressure and hypertension Stage 1 and Stage >or=2 were 18.0%, 20.1%, and 10.1% for men and 15.9%, 15.6%, and 8.8% for women, respectively. Antihypertensive drug users were also classified into the baseline blood pressure categories. Compared with the optimal blood pressure group, the multivariable hazard ratios (95% confidence intervals) of CVD for normal and high-normal blood pressure and hypertension Stage 1 and Stage >or=2 were 2.04 (1.19 to 3.48), 2.46 (1.46 to 4.14), 2.62 (1.59 to 4.32), and 3.95 (2.37 to 6.58) in men and 1.12 (0.59 to 2.13), 1.54 (0.85 to 2.78), 1.35 (0.75 to 2.43), and 2.86 (1.60 to 5.12) in women, respectively. The risks of myocardial infarction and stroke for each blood pressure category were similar to those of CVD. Population-attributable fractions of high-normal blood pressure and hypertension for CVD were 12.2% and 35.3% in men and 7.1% and 23.4% in women, respectively. In conclusion, high-normal blood pressure is a risk factor for the incidence of stroke and myocardial infarction in a general urban population of Japanese men.


A comparative analysis of mycotoxin contamination of supermarket and premium brand pelleted dog food in Durban, South Africa.
Dry pelleted dog food in the South African market is available via supermarkets, pet stores (standard brands [SBs]) and veterinary channels (premium brands [PBs]). For the purpose of this study, the supermarket channel included the cheaper quality foods and PBs were sold via the veterinary channel (n = 20). These feeds were analysed for four main mycotoxins (aflatoxins [AF], fumonisin [FB], ochratoxin A [OTA] and zearalenone [ZEA]) using standard welldescribed extraction, characterisation and quantitation processes. Irrespective of the brand or marketing channel, all foods were contaminated with fungi (mainly Aspergillus flavus, Aspergillus fumigatus and Aspergillus parasiticus) and mycotoxins (most prevalent being aflatoxins and fumonisins). This was observed in all 20 samples irrespective of the marketing channel or perceived quality. Also, many samples within each marketing channel failed the 10 ppb limit for aflatoxin set by regulations in South Africa. Although fumonisin was detected in all samples, a single sample failed the Food and Drug Administration (FDA) limit of 100 ppb. Both OTA and ZEA were found at low concentrations and were absent in some samples. This study suggested that higher priced dog food does not ensure superior quality or that it is free from contamination with fungi or mycotoxins. However, analysis of the more expensive PBs did reveal contamination concentrations lower than those of the SBs.


Fomes fomentarius and Tricholoma anatolicum (Agaricomycetes) Extracts Exhibit Significant Multiple Drug-Resistant Modulation Activity in Drug-Resistant Breast Cancer Cells.
Multiple drug resistance is one of the main problems that hinder successful cancer chemotherapy. Investigations on the development of effective chemotherapeutic agents and drug resistance inhibitors motivate studies on the effects of natural compounds on drug-resistant cancer cells. For this purpose, aqueous, methanol, and ethanol extracts of Fomes fomentarius and Tricholoma anatolicum were prepared. The extracts were evaluated to assess their anticancer and multiple drug resistance modulation activities. Cytotoxic effects of F. fomentarius and T. anatolicum extracts on paclitaxel and vincristine resistant P-glycoprotein over-expressing MCF-7 cell lines were investigated by cytotoxicity test (XTT). P-glycoprotein reversing ability and MDR modulation effects of the extracts were determined by flow cytometry through Rhodamine 123 exclusion assay. Furthermore, 11 phenolic compounds in the extracts were characterized by HPLC. As a result of the cytotoxicity assay, IC50 values of the extracts for MCF-7/Vinc were between 1.08 and 1.80 mg/mL, and IC50 values for MCF-7/Pac were found between 1.11 and 2.83 mg/mL. Strikingly, methanol extract of F. fomentarius and ethanol extract of T. anatolicum have potential value to become MDR reversing agents for drug-resistant breast cancer cells.


Expression of membrane-associated proteins within single emulsion cell facsimiles.
MreB is a structural membrane-associated protein which is one of the key components of the bacterial cytoskeleton. Although it plays an important role in shape maintenance of rod-like bacteria, the understanding of its mechanism of action is still not fully understood. This study shows how segmented flow and microdroplet technology can be used as a new tool for biological in vitro investigation of this protein. In this paper, we demonstrate cell-free expression in a single emulsion system to express red fluorescence protein (RFP) and MreB linked RFP (MreB-RFP). We follow the aggregation and localisation of the fusion protein MreB-RFP in this artificial cell-like environment. The expression of MreB-RFP in single emulsion droplets leads to the formation of micrometer-scale protein patches distributed at the water/oil interface.


Effect of pathogenic yeasts on human platelet aggregation.
We investigated the effects of Candida albicans, Cryptococcus neoformans and the respective culture supernatants on human platelet aggregation (PA). Both yeasts were unable to aggregate the platelets directly. On the other hand, cells of these yeasts significantly (P < 0.01) inhibited PA at concentrations equal to or higher than 1 x 10(6) cells/mL for C. albicans and equal to or higher than 1 x 10(5) cells/mL for C. neoformans. When the supernatants of one-week broth cultures were added to the activated platelets no inhibition in aggregation was observed. Apparently somatic components of these yeasts, but not their metabolic products, exert an antagonistic effect on the aggregation of human platelets, possibly aiding the fungi in their evasion of the microbicidal defense system during vascular dissemination.


Tat 101-mediated enhancement of brain pericyte migration involves platelet-derived growth factor subunit B homodimer: implications for human immunodeficiency virus-associated neurocognitive disorders.
In the era of antiretroviral therapy, although the human immunodeficiency virus (HIV) replication can be successfully controlled, complications of the CNS continue to affect infected individuals. Viral Tat protein is not only neurotoxic but has also been shown to disrupt the integrity of the blood-brain barrier (BBB). Although the role of brain microvascular endothelial cells and astrocytes in Tat-mediated impairment has been well documented, pericytes, which are important constituents of the BBB and play a key role in maintaining the integrity of the barrier, remain poorly studied in the context of HIV-associated neurocognitive disorders (HAND). In the present study, we demonstrated that exposure of human brain microvascular pericytes and C3H/10T1/2 cells to HIV-1 Tat101 resulted in increased expression of platelet-derived growth factor subunit B homodimer (PDGF-BB) and increased migration of the treated cells. Furthermore, we also demonstrated that this effect of Tat was mediated via activation of mitogen-activated protein kinases and nuclear factor-κB pathways. Secreted PDGF-BB resulted in autocrine activation of the PDGF-BB/PDGF β receptor signaling pathway, culminating ultimately into increased pericyte migration. Ex vivo relevance of these findings was further corroborated in isolated microvessels of HIV Tg26 mice that demonstrated significantly increased expression of PDGF-BB in isolated brain microvessels with a concomitant loss of pericytes. Intriguingly, loss of pericyte coverage was also detected in sections of frontal cortex from humans with HIV-encephalitis compared with the uninfected controls. These findings thus implicate a novel role of PDGF-BB in the migration of pericytes, resulting in loss of pericyte coverage from the endothelium with a subsequent breach of the BBB.


Scheduled administration of low dose irinotecan before gemcitabine in the second line therapy of non-small cell lung cancer: a phase II study.
We had previously demonstrated that low dose irinotecan (CPT-11) leads to increased accumulation of cells in S-phase and shows a therapeutic synergy with S-phase specific chemotherapy such as gemcitabine and 5-fluorouracil. In this phase II study, our objectives were to evaluate the tolerability and activity of low dose CPT-11 followed 24 h later by gemcitabine as second line therapy in patients with metastatic non-small cell lung cancer (NSCLC). CPT-11 (60 mg/m) was administered 24 h before gemcitabine (1000 mg/m) on days 1, 2, 8, and 9 every 3 weeks. Twenty-nine patients were evaluable for response. The median follow-up was 7.4 months. Partial response (PR) was seen in two (6.9, 95% confidence interval (CI): 0.009-0.228). PR and stable disease were seen in 22 patients (75.9, 95% CI: 0.564-0.897). The median survival time was 13.8 months (95% CI: 8.1-19.3). The median time to progression was 4.6 months (95% CI: 2.6-6.2). Thirty-eight patients were evaluable for toxicity. Neutropenia (grade 3 or 4) was observed in 27 patients (71%). Eight patients did not receive cycle 2 of therapy owing to prolonged neutropenia. No treatment-related deaths occurred. Scheduled administration of low dose CPT-11, 24 h before gemcitabine in the second line therapy of NSCLC yielded comparable disease control rates (PR and stable disease) when compared with other studies using the two chemotherapy drugs in the traditional sequence. However, this approach was associated with higher grade 3/4 neutropenia and is not recommended for further study in metastatic NSCLC.


Trans-synaptic increase in RNA coding for tyrosine hydroxylase in a rat sympathetic ganglion.
To begin examining molecular mechanisms underlying trans-synaptic regulation, tyrosine hydroxylase (TH) and its messenger RNA (mRNA) were examined in the superior cervical sympathetic ganglion (SCG) of adult rats. Basal levels of TH mRNA were detectable in control ganglia by RNA dot hybridization, using the 32P nick translated PstI-KpnI restriction fragment of pTH.4 as a probe. Reserpine induced a 3-fold rise in TH activity per microgram protein, and a simultaneous 3-fold increase in ganglion TH mRNA. As expected, ganglion decentralization (denervation) prevented the trans-synaptic induction of TH. In addition, decentralization prevented the increase in TH mRNA, suggesting that the increase in message was dependent on trans-synaptic stimulation. Northern blot analysis indicated that the cDNA (complementary DNA) probe hybridized to a single band of approximately 1900 nucleotides, which was the same size in all ganglia. Our observations indicate that induction of TH is associated with a trans-synaptic increase in mRNA coding for the enzyme. Consequently, trans-synaptic increases in impulse activity may induce TH by increasing neuronal levels of TH mRNA in the SCG.