Signature of VDR miRNAs and epigenetic modulation of vitamin D signaling in melanoma cell lines.
Melanoma cells express the nuclear vitamin D receptor (VDR), indicating that malignant melanoma represents a promising target for treatment with 1,25-dihydroxyvitamin D3 (1,25(OH)(2)D(3)) or its analogs. We previously showed that some melanoma cell lines are resistant to the antiproliferative effects of 1,25(OH)(2)D(3) and that 1,25(OH)(2)D(3)-sensitivity can, at least in part, be restored by co-treatment with the histone deacetylase inhibitor (HDACI) Trichostatin A (TSA) or with the DNA methyltransferase inhibitor (DNMTI), 5-azacytidine (5-Aza). This study aimed at gaining further insights into the molecular mechanisms that underlie the epigenetic modulation of 1,25(OH)(2)D(3)-sensitivity in melanoma cells. The expression of VDR mRNA, protein and two candidates of VDR microRNAs (miR-125b, miR-27b) were compared in 1,25(OH)(2)D(3)-responsive (MeWo, SK-Mel28) and -resistant (SK-Mel5, IGR) melanoma cell lines and in normal human melanocytes (NHM) using real time PCR and western blot analysis. Additionally, the effect of 1,25(OH)(2)D(3), epigenetic modulating drugs (TSA, 5-Aza) and miR-125b antisense on the expression of VDR messenger RNA (mRNA)/protein, miR-125b and miR-27b was investigated. Treatment with 1,25(OH)(2)D(3) and/or epigenetic drugs (5-Aza, TSA) modulated the VDR mRNA expression in the 1,25(OH)(2)D(3)-responsive and - resistant melanoma cell lines and in the NHM. Treatment with 5-Aza, but not with TSA, reduced the expression of miR-125b in the 1,25(OH)(2)D(3)-responsive and -resistant melanoma cell lines and in the NHM. Treatment with 1,25(OH)(2)D(3) and/or epigenetic drugs (5-Aza, TSA) reduced the miR-27b expression in three out of four melanoma cell lines. Moreover, no difference was observed in VDR protein expression in the 1,25(OH)(2)D(3)-responsive as compared to the 1,25(OH)(2)D(3)-resistant melanoma cell lines. Transfection with miR-125b antisense did not affect the VDR mRNA/protein expression in the IGR cells. Responsiveness to 1,25(OH)(2)D(3) corresponds to the expression level of VDR mRNA which in turn might be regulated by VDR microRNAs or epigenetic modulating drugs.


Some reflex cardioinhibitory responses in the cat and their modulation by central inspiratory neuronal activity.
1. Cats were anaesthetized with a mixture of chloralose and urethane, and were artifically ventilated. 2. An open pneumothorax was provided by two large-bore tubes which were sealed in the sixth intercostal space on each side. They were connected to a Fleisch pneumotachograph. Phasic changes in central inspiratory neuronal activity were measured quantitatively as changes in the volume of the pneumothorax during temporary interruption of artificial respiration, the volume of the lungs being held constant at their end-expiratory level. In this way the activity of slowly adapting pulmonary stretch receptors was maintained constant. 3. Reflex cardioinhibitory responses were elicited by stimulation of (a) the carotid body chemoreceptors by intracarotid injections of cyanide; (b) the arterial baroreflex by controlled elevations of the blood pressure; (c) cardiac receptors by left atrial injections of veratridine; and (d) pulmonary C fibres (including J receptors) by right atrial injections of phenylbiguanide. 4. The effects of central inspiratory neuronal activity on pulse interval were assessed by comparing the values observed during the inspiratory and expiratory phases of the respiratory cycle in the control state and during stimulation of each cardiovascular receptor group. 5. The carotid chemoreceptor-induced bradycardia measured during the expiratory phase of respiration was reduced during inspiration to a value of about 15% of control. The central inspiratory drive was less effective in altering the reflex responses from the arterial baroreceptors and cardiac receptors, the corresponding values being 42 and 51% respectively. 6. In contrast, the bradycardia evoked by pulmonary C fibre stimulation was not significantly affected by the central inspiratory drive. 7. The differential nature of the modulation by the central inspiratory drive occurred independently of the integrity of the sympathetic nerve supply to the heart indicating that the cardiac efferents involved were largely fibres in the vagus nerves. 8. The possible explanation of these results in terms of central mechanisms is discussed.


Akt inhibition promotes hexokinase 2 redistribution and glucose uptake in cancer cells.
Hexokinase II (HK2), the enzyme that catalyzes the first committed step of glycolysis, is overexpressed in many cancers, as is the central signaling kinase Akt. Akt activity promotes HK2 association with the mitochondria, as well as glucose uptake by cancer cells. In HeLa cervical cancer cells, Akt inhibitor IV (Ai4) increased nuclear HK2 localization, while in MDA-MB-231 breast cancer cells, Ai4 merely induced cytoplasmic redistribution without increased nuclear accumulation. Small interfering RNA (siRNA) directed against Akt confirmed the effect in HeLa cells. Next, we treated the cells with clotrimazole (CTZ), which detaches HK2 from the mitochondria, or leptomycin B (LMB), which promotes HK2 nuclear accumulation, and determined the effect on HK2 subcellular distribution. In both cell lines, CTZ detached HK2 from the mitochondria, without substantially increasing nuclear HK2, while LMB increased nuclear HK2, without redistributing cytoplasmic HK2. Contrary to expectations, Akt inhibition promoted glucose uptake in both cell lines, suggesting that Akt inhibition may increase glucose uptake by detaching HK2 from the mitochondria. In both cell lines, CTZ and LMB increased glucose uptake. However, the results in the HeLa cells showed greater effects: CTZ increased glucose uptake to a similar degree to Ai4, while LMB was far more effective than either. These data suggest that both detachment of HK2 from the mitochondria and increased nuclear HK2 are important for Ai4-induced increased glucose uptake.


Autoantibodies from synovial lesions in chronic, antibiotic treatment-resistant Lyme arthritis bind cytokeratin-10.
Although the causative agent of Lyme disease is definitively known to be the tick-borne spirochete, Borrelia burgdorferi, the etiology of chronic joint inflammation that ensues in a subset of patients remains less well understood. Persistence of arthritis after apparent eradication of the spirochete suggests an autoimmune reaction downstream of the original bacterial infection. We have generated recombinant Ab probes from synovial lesions within affected arthritic joints in an attempt to recapitulate disease-relevant Ag-binding specificities at the site of injury. Using this panel of intra-articular probes, as well as Ab fragments derived from patient peripheral blood, we have identified cytokeratin 10, present in synovial microvascular endothelium, as a target ligand and a putative autoantigen in chronic, antibiotic treatment-resistant Lyme arthritis. Furthermore, there is cross-reactivity between cytokeratin 10 and a prominent B. burgdorferi Ag, outer surface protein A. Release of the self protein in the context of inflammation-induced tissue injury and the resulting in situ response to it could set in motion a feed-forward loop, which amplifies the inflammatory process, thereby rendering it chronic and self-perpetuating, even in the absence of the inciting pathogen.


Women's experiences with ovarian cancer: reflections on being diagnosed.
The purpose of this qualitative study was to describe the perspectives of women living with ovarian cancer about their experiences with diagnosis, treatment, and follow-up care. A convenience sample of 18 women was interviewed using an open-ended interview guide. This paper will focus on their perspectives during the peri-diagnostic period. Most of the women in this study experienced changes in their bodies--bloating, weight gain around their middles, indigestion, bowel changes, and abdominal pain. The vagueness of the symptoms and lack of awareness by the women and, in the women's opinion, health care professionals caused delays in diagnosis and initial investigations of other body systems. The symptoms were often dismissed by the women as being related to normal body changes such as childbirth, menopause, or stress responses. It was often not until the women's symptoms became severe or included pain that referrals were made to relevant specialists. At this point, many of the women were physically and emotionally spent, leaving them vulnerable and in a weakened state for confronting the diagnosis and coping with the difficult course of treatment. Clearly, the subtle, nebulous nature of the symptoms confounded the diagnosis of ovarian cancer for these women, resulting in patient delays and physician delays.


Usefulness of Cardiovascular Magnetic Resonance Imaging in a Patient with Cardiac Involvement of Systemic Sclerosis.
A 72-year-old Japanese woman with systemic sclerosis was admitted to our hospital because of symptoms of heart failure. Cardiovascular magnetic resonance (CMR) imaging had shown that extensive myocardial fibrosis secondary to systemic sclerosis was the main cause of heart failure. One month after CMR, she had complete atrioventricular (AV) block. It was suggested that the progression of fibrosis to the AV node caused complete AV block. This case report has clinical implications in highlighting the fact that CMR is useful for not only evaluating the present pathophysiology but also predicting future adverse events in patients with systemic sclerosis.


Ertapenem-Induced Encephalopathy in a Patient With Normal Renal Function.
Drug-induced neurotoxicity is a rare adverse reaction associated with ertapenem. Encephalopathy is a type of neurotoxicity that is defined as a diffuse disease of the brain that alters brain function or structure. We report a patient with normal renal function who developed ertapenem-induced encephalopathy manifesting as altered mental status, hallucinations, and dystonic symptoms. The patient's symptoms improved dramatically following ertapenem discontinuation, consistent with case reports describing ertapenem neurotoxicity in renal dysfunction. Since clinical evidence strongly suggested ertapenem causality, we utilized the Naranjo Scale to estimate the probability of an adverse drug reaction to ertapenem. Our patient received a Naranjo Scale score of 7, suggesting a probable adverse drug reaction, with a reasonable temporal sequence to support our conclusion.


Microenvironment-associated gene HSD11B1 may serve as a prognostic biomarker in clear cell renal cell carcinoma: a study based on TCGA, RT‑qPCR, Western blotting, and immunohistochemistry.
Clear cell renal cell carcinoma (ccRCC) is one of the most common malignant tumors worldwide. The clinical treatment of ccRCC is strongly associated with the tumor microenvironment (TME). Identifying potential markers of ccRCC is important to improve prognosis. Therefore, in the present study, the levels of immune/stromal components and the proportion of tumor-infiltrating immune cells (TIICs) were determined in 611 ccRCC samples using the ESTIMATE and CIBERSORT analytical tools. Subsequently, hydroxysteroid 11-beta dehydrogenase-1 (HSD11B1) was identified by univariate Cox regression analysis, protein-protein interaction (PPI) networks and clinical survival analysis to be associated with ccRCC prognosis. At the same time, the abundance of HSD11B1 increased significantly in ccRCC was verified by western blotting, RT‑qPCR and immunostaining analysis. Furthermore, Gene Set Enrichment Analysis (GSEA) and TME suggested that HSD11B1 was involved in TME immune-related status. Taken together, the results of the present study demonstrated that HSD11B1 is a potential prognostic biomarker associated with immune cell infiltration in ccRCC.


GourmetNet: Food Segmentation Using Multi-Scale Waterfall Features with Spatial and Channel Attention.
We propose GourmetNet, a single-pass, end-to-end trainable network for food segmentation that achieves state-of-the-art performance. Food segmentation is an important problem as the first step for nutrition monitoring, food volume and calorie estimation. Our novel architecture incorporates both channel attention and spatial attention information in an expanded multi-scale feature representation using our advanced Waterfall Atrous Spatial Pooling module. GourmetNet refines the feature extraction process by merging features from multiple levels of the backbone through the two attention modules. The refined features are processed with the advanced multi-scale waterfall module that combines the benefits of cascade filtering and pyramid representations without requiring a separate decoder or post-processing. Our experiments on two food datasets show that GourmetNet significantly outperforms existing current state-of-the-art methods.


The Relationship between Fungal Diversity and Invasibility of a Foliar Niche-The Case of Ash Dieback.
European ash (Fraxinus excelsior) is threatened by the invasive ascomycete Hymenoscyphus fraxineus originating from Asia. Ash leaf tissues serve as a route for shoot infection but also as a sporulation substrate for this pathogen. Knowledge of the leaf niche partitioning by indigenous fungi and H. fraxineus is needed to understand the fungal community receptiveness to the invasion. We subjected DNA extracted from unwashed and washed leaflets of healthy and diseased European ash to PacBio sequencing of the fungal ITS1-5.8S-ITS2 rDNA region. Leaflets from co-inhabiting rowan trees (Sorbus aucuparia) served as a reference. The overlap in leaflet mycobiomes between ash and rowan was remarkably high, but unlike in rowan, in ash leaflets the sequence read proportion, and the qPCR-based DNA amount estimates of H. fraxineus increased vigorously towards autumn, concomitant with a significant decline in overall fungal richness. The niche of ash and rowan leaves was dominated by epiphytic propagules (Vishniacozyma yeasts, the dimorphic fungus Aureobasidion pullulans and the dematiaceous hyphomycete Cladosporium ramotenellum and H. fraxineus), and endophytic thalli of biotrophs (Phyllactinia and Taphrina species), the indigenous necrotroph Venturia fraxini and H. fraxineus. Mycobiome comparison between healthy and symptomatic European ash leaflets revealed no significant differences in relative abundance of H. fraxineus, but A. pullulans was more prevalent in symptomatic trees. The impacts of host specificity, spatiotemporal niche partitioning, species carbon utilization profiles and life cycle traits are discussed to understand the ecological success of H. fraxineus in Europe. Further, the inherent limitations of different experimental approaches in the profiling of foliicolous fungi are addressed.


Breathers in a system with helicity and dipole interaction.
Recent papers that have studied variants of the Peyrard-Bishop model for DNA, have taken into account the long range interaction due to the dipole moments of the hydrogen bonds between base pairs. In these models the helicity of the double strand is not considered. In this paper we have performed an analysis of the influence of the helicity on the properties of static and moving breathers in a Klein-Gordon chain with dipole-dipole interaction. It has been found that the helicity enlarges the range of existence and stability of static breathers, although this effect is small for a typical helical structure of DNA. However, the effect of the orientation of the dipole moments is considerably higher with transcendental consequences for the existence of mobile breathers.


Cancer risk assessment, indicators, and guidelines for polycyclic aromatic hydrocarbons in the ambient air.
Polycyclic aromatic hydrocarbons (PAHs) are formed during incomplete combustion. Domestic wood burning and road traffic are the major sources of PAHs in Sweden. In Stockholm, the sum of 14 different PAHs is 100-200 ng/m(3) at the street-level site, the most abundant being phenanthrene. Benzo[a]pyrene (B[a]P) varies between 1 and 2 ng/m(3). Exposure to PAH-containing substances increases the risk of cancer in humans. The carcinogenicity of PAHs is associated with the complexity of the molecule, i.e., increasing number of benzenoid rings, and with metabolic activation to reactive diol epoxide intermediates and their subsequent covalent binding to critical targets in DNA. B[a]P is the main indicator of carcinogenic PAHs. Fluoranthene is an important volatile PAH because it occurs at high concentrations in ambient air and because it is an experimental carcinogen in certain test systems. Thus, fluoranthene is suggested as a complementary indicator to B[a]P. The most carcinogenic PAH identified, dibenzo[a,l]pyrene, is also suggested as an indicator, although it occurs at very low concentrations. Quantitative cancer risk estimates of PAHs as air pollutants are very uncertain because of the lack of useful, good-quality data. According to the World Health Organization Air Quality Guidelines for Europe, the unit risk is 9 X 10(-5) per ng/m(3) of B[a]P as indicator of the total PAH content, namely, lifetime exposure to 0.1 ng/m(3) would theoretically lead to one extra cancer case in 100,000 exposed individuals. This concentration of 0.1 ng/m(3) of B[a]P is suggested as a health-based guideline. Because the carcinogenic potency of fluoranthene has been estimated to be approximately 20 times less than that of B[a]P, a tentative guideline value of 2 ng/m(3) is suggested for fluoranthene. Other significant PAHs are phenanthrene, methylated phenanthrenes/anthracenes and pyrene (high air concentrations), and large-molecule PAHs such as dibenz[a,h]anthracene, benzo[b]fluoranthene, benzo[k]fluoranthene, and indeno[1,2,3-cd]pyrene (high carcinogenicity). Additional source-specific indicators are benzo[ghi]perylene for gasoline vehicles, retene for wood combustion, and dibenzothiophene and benzonaphthothiophene for sulfur-containing fuels.


Data uncertainties in material flow analysis: Municipal solid waste management system in Maputo City, Mozambique.
Material flow analysis can effectively trace and quantify the flows and stocks of materials such as solid wastes in urban environments. However, the integrity of material flow analysis results is compromised by data uncertainties, an occurrence that is particularly acute in low-and-middle-income study contexts. This article investigates the uncertainties in the input data and their effects in a material flow analysis study of municipal solid waste management in Maputo City, the capital of Mozambique. The analysis is based on data collected in 2007 and 2014. Initially, the uncertainties and their ranges were identified by the data classification model of Hedbrant and Sörme, followed by the application of sensitivity analysis. The average lower and upper bounds were 29% and 71%, respectively, in 2007, increasing to 41% and 96%, respectively, in 2014. This indicates higher data quality in 2007 than in 2014. Results also show that not only data are partially missing from the established flows such as waste generation to final disposal, but also that they are limited and inconsistent in emerging flows and processes such as waste generation to material recovery (hence the wider variation in the 2014 parameters). The sensitivity analysis further clarified the most influencing parameter and the degree of influence of each parameter on the waste flows and the interrelations among the parameters. The findings highlight the need for an integrated municipal solid waste management approach to avoid transferring or worsening the negative impacts among the parameters and flows.


Preclinical disposition of MGS0274 besylate, a prodrug of a potent group II metabotropic glutamate receptor agonist MGS0008 for the treatment of schizophrenia.
MGS0274 besylate is an ester-based lipophilic prodrug of a metabotropic glutamate (mGlu) 2 and mGlu3 receptor agonist MGS0008 and being developed for the treatment of schizophrenia. We investigated the disposition of these compounds in rats and monkeys and in vitro metabolism in humans to evaluate whether MGS0274 besylate could be useful as a prodrug in humans. After the oral administration of MGS0274 besylate to monkeys (2.89 mg/kg), MGS0008 was immediately found in plasma, reached a maximum concentration at 4 hours postdose, and decreased with a terminal half-life of 16.7 hours; MGS0274 was barely detectable. The oral bioavailability as MGS0008 was 83.7%, which was approximately 20-fold greater than that after oral dosing of MGS0008 (3.8%). In rats, MGS0008 penetrated the cerebrospinal fluid and was eliminated slower than from plasma. The in vitro metabolism study indicated that MGS0274 was rapidly hydrolyzed to MGS0008, which was not further metabolized. After the intravenous administration of MGS0008 to rats and monkeys, almost all the dose was excreted unchanged in urine. These results suggested that MGS0274 was, as expected, presystemically hydrolyzed to MGS0008 after gastrointestinal absorption and that MGS0008 was distributed throughout the body without further metabolism and ultimately excreted in urine in the animals. Furthermore, the hydrolytic activity against MGS0274 in the human liver S9 fraction was comparable to that in monkeys, suggesting the possibility of the rapid presystemic hydrolysis of MGS0274 to MGS0008 in humans, as it is in monkeys. Consequently, MGS0274 besylate is expected to function as a preferable prodrug in humans.


Antitumor triptycene analogs induce a rapid collapse of mitochondrial transmembrane potential in HL-60 cells and isolated mitochondria.
Since synthetic analogs of triptycene (TT code number), such as bisquinones TT2 and TT13, can trigger cytochrome c release without caspase activation and retain their ability to induce apoptosis in multidrug-resistant (MDR) tumor cells, fluorescent probes of transmembrane potential have been used to determine whether these antitumor compounds might directly target mitochondria in cell and cell-free systems to cause the collapse of mitochondrial membrane potential ( downward arrow Deltapsim) that is linked to permeability transition pore (PTP) opening. Using JC-1 dye, the abilities of various TT analogs to induce the downward arrow Deltapsim in wild-type and MDR HL-60 cells are rapid (within 5-20 min), irreversible after drug removal, concentration dependent in the 0.64-25 microM range, and generally related to their antitumor activities in vitro. The downward arrow Deltapsim caused by TT2 and TT13, which are more potent than mitoxantrone, staurosporine and the reference depolarizing agent, carbonyl cyanide m-chlorophenylhydrazone (CCCP), in HL-60 cells, are not prevented by caspase-2 or -8 inhibitors, suggesting that activation of these apical caspases upstream of mitochondria is not involved in this process. Antitumor TT analogs (0.64-25 microM) also mimic the abilities of the known depolarizing agents, CCCP, alamethicin, gramicidin A and 100 microM CaCl(2), to directly induce within 20 min the downward arrow Deltapsim in isolated mitochondria prepared from mouse liver and loaded with rhodamine 123 dye. The fact that 20 microM Ca(2+), which is insufficient to trigger depolarization on its own, is required to reveal the depolarizing effect of TT2 in isolated mitochondria suggests that antitumor TT analogs might interact with the PTP to alter its conformation and increase its Ca(2+) sensitivity. Indeed, such Ca(2+)-dependent downward arrowDeltapsim of isolated mitochondria treated with 25 microM TT2 or 100 microM Ca(2+) are blocked by ruthenium red. Daunorubicin (DAU) is unable to mimic the rapid downward arrowDeltapsim caused by antitumor TT bisquinones within 5-40 min of treatment in HL-60 cells or isolated mitochondria. Moreover, the downward arrowDeltapsim caused by 25 microM TT2 or 100 microM Ca(2+) in isolated mitochondria are similarly blocked by cyclosporin A (CsA), bongkrekic acid and decylubiquinone, which prevent PTP opening, suggesting that, in contrast to DAU, antitumor TT analogs that directly target mitochondria to trigger the Ca(2+)-dependent and CsA-sensitive downward arrowDeltapsim, might induce PTP opening and the mitochondrial pathway of apoptosis even in the absence of nuclear signals.


Retinopathy of prematurity in Saudi Arabia: Exploring maternal risk factors.
Retinopathy of prematurity (ROP) is a preventable blinding disorder affecting preterm infants. To date, maternal risk factors have not been studied in Saudi Arabia. This study aims to identify possible maternal risk factors for any stage and type 1 ROP. A total of 295 preterm infants screened for ROP between November 2013 and December 2018 at a Saudi Arabian tertiary-care hospital were included. We included infants with a gestational age ≤ 32 weeks and/or birth weight (BW) ≤1500 g. We analyzed 28 maternal and neonatal risk factors. The incidence of ROP at any stage and Type 1 were 31.9% and 7%, respectively. In the univariate analysis, the only maternal factor associated with any stage of ROP was spontaneous vaginal delivery (P = 0.049), but no maternal factor was an independent risk factor for type 1 ROP. Multivariate logistic regression analysis identified lower BW, lower gestational age and longer neonatal intensive care unit stay as independent risk factors for the development of ROP at any stage (P < 0.05). For Type 1 ROP, lower BW, and intraventricular hemorrhage were significant independent risk factors (P < 0.05). The only maternal risk factor related to ROP was spontaneous vaginal delivery, which increased the risk of any stage of ROP. The single most predictive risk factor for any stage of ROP and Type 1 ROP was low BW. These findings emphasize the role of the obstetrician in promoting health care and modifying maternal risk factors to prevent preterm births related to a low BW.


Affinity purification of ribosomes to access the translatome.
We describe ribosome affinity purification (RAP), a method that allows rapid purification of ribosomes and associated messages from the yeast Saccharomyces cerevisiae. The method relies on the expression of protein A tagged versions of the ribosomal protein Rpl16, which is used to efficiently recover endogenously formed ribosomes and polysomes from cellular extracts with IgG-coupled spherical microbeads. This approach can be applied to profile reactions of the translatome, which refers to all messages associated with ribosomes, with those of the transcriptome using DNA microarrays. In addition, ribosomal proteins, their modifications, and/or other associated proteins can be mapped with mass spectrometry. Finally, application of this method in other organisms provides a valuable tool to decipher cell-type specific gene expression patterns.


Role of peroxiredoxin III in the pathogenesis of pre-eclampsia as evidenced in mice.
As a member of peroxiredoxin (Prx) family, PrxIII has been demonstrated to play an important role in scavenging intracellular reactive oxygen species (ROS). Since PrxIII knockout mice exhibited oxidative stress in placentas resembling pathophysiologic changes in placentas of human pre-eclampsia, we measured blood pressure through the carotid artery and detected oxidative status by Western blotting in pregnant mice. We did not notice hypertension in pregnant PrxIII knockout mice as compared with wild-type littermates, although endothelin-1 was over-expressed in PrxIII-deficient placentas. Our results indicate that PrxIII is not involved in pre-eclamptic development. Instead, PrxIII is an indispensable antioxidant in placentas where oxidative stress exists.


Activation of Poly(ADP-Ribose)Polymerase in rat hepatocytes does not contribute to their cell death by oxidative stress.
Oxidative stress induced by tert-butyl hydroperoxide (tBOOH) in freshly isolated rat hepatocytes caused DNA damage and loss of membrane integrity. Such DNA lesions are likely to be single strand breaks since neither caryolysis nor chromatine condensation was seen in electron micrographs from tBOOH-treated cells. In addition, pulsed field gel electrophoresis of genomic DNA from both control and tBOOH-treated hepatocytes showed similar profiles, indicating the absence of internucleosomal DNA cleavage, a classical reflection of apoptotic endonuclease activity. The activation of the repair enzyme poly(ADP-ribose)polymerase (PARP) following DNA damage by tBOOH induced a dramatic drop in both NAD(+) and ATP. The inhibition of PARP by 3-aminobenzamide enhanced DNA damage by tBOOH, restored NAD(+) and ATP levels, but did not result in better survival against cell killing by tBOOH. The lack of the protective effect of PARP inhibitor, therefore, does not implicate PARP in the mechanism of tBOOH-induced cytotoxicity. Electron micrographs also show no mitochondrial swelling in cells under oxidative stress, but such organelles were mainly located around the nucleus, a picture already observed in autoschizis, a new suggested kind of cell death which shows both apoptotic and necrotic morphological characteristics.


Natural polymorphisms in a pair of NSP2 homoeologs can cause loss of nodulation in peanut.
Microbial symbiosis in legumes is achieved through nitrogen-fixing root nodules, and these are important for sustainable agriculture. The molecular mechanisms underlying development of root nodules in polyploid legume crops are largely understudied. Through map-based cloning and QTL-seq approaches, we identified a pair of homoeologous GRAS transcription factor genes, Nodulation Signaling Pathway 2 (AhNSP2-B07 or Nb) and AhNSP2-A08 (Na), controlling nodulation in cultivated peanut (Arachis hypogaea L.), an allotetraploid legume crop, which exhibited non-Mendelian and Mendelian inheritance, respectively. The segregation of nodulation in the progeny of Nananbnb genotypes followed a 3:1 Mendelian ratio, in contrast to the 5:3~1:1 non-Mendelian ratio for nanaNbnb genotypes. Additionally, a much higher frequency of the nb allele (13%) than the na allele (4%) exists in the peanut germplasm collection, suggesting that Nb is less essential than Na in nodule organogenesis. Our findings reveal the genetic basis of naturally occurred non-nodulating peanut plants, which can be potentially used for nitrogen fixation improvement in peanut. Furthermore, the results have implications for and provide insights into the evolution of homoeologous genes in allopolyploid species.


A confirmed case of Rickettsia parkeri infection in a traveler from Uruguay.
The first confirmed case of Rickettsia parkeri infection in Uruguay is reported. To date, in South America, molecularly confirmed cases of human infection have been found in Argentina and probably, Brazil. Our patient returned to Spain after a 7-day trip to Colonia Suiza (Southwestern Uruguay). He presented fever (39°C), chills, and two eschars (tache noire-like) surrounded by an indurated, erythematous halo on the inner side of the left ankle besides a maculopapular rash on the legs. After treatment with doxycycline for 7 days, he fully recovered. R. parkeri infection was diagnosed by molecular-based detection of the microorganism in a swab specimen of the eschar. Diagnosis was supported by seroconversion between acute- and convalescent-phase sera specimens.


Tumor control and morbidity following transperineal iodine 125 implantation for stage T1/T2 prostatic carcinoma.
To quantify disease progression and morbidity following computer tomography (CT)-based transperineal iodine 125 prostate implantation. Ninety-two patients with clinical stage T1 or T2, Gleason score 2 to 7/10, prostatic carcinoma had outpatient, CT-based transperineal 125I prostate implantation and were monitored for 1 to 7 years (median, 3). The prescribed minimum radiation dose was 140 to 160 Gy. Lymph node dissection and postimplantation prostatic biopsies were not routinely performed. In 46% of patients, radiation-related urinary symptoms were substantial enough at 1 month following implantation to require medication. Radiation-related urinary symptoms gradually resolved. Two years after implantation, 14% of patients had persistent urinary symptoms of Radiation Therapy Oncology Group (RTOG) > or = grade 2. Eight percent of patients underwent a transurethral resection of the prostate (TURP) within 2 years of implantation. Five patients developed radiation-induced rectal ulcerations. Of 56 patients who were sexually potent preimplantation, 86% retained potency at 3 years. Twenty-five patients had biochemical disease progression. The overall actuarial freedom from biochemical failure rate at 4 years following implantation was 63%. In Cox proportional hazards multivariate analysis, the strongest predictor of failure was prostate-specific antigen (PSA) level less than or greater than 10 ng/mL (P = .005), followed by Gleason score (2 to 4 v 5 to 7, P = .08) and stage (T1 v T2, P = .09). The 5-year biochemical freedom-from-progression rates following transperineal 125I implantation are comparable with those achieved with prostatectomy. The morbidity has decreased with increased physician experience.


Characteristics of physicochemical adsorption of soluble matter by particles formed in a fluidized pellet bed reactor.
To investigate the mechanism of soluble matter removal by a fluidized pellet bed (FPB) reactor, an experimental study was conducted using a laboratory-scale FPB device for treating synthetic wastewater under a condition of no activated sludge seeding and no dissolved oxygen supply so that the physicochemical functions of the FPB reactor could be evaluated. By using polyaluminium chloride and polyacrylamide as coagulants, it was found that most of the substances in the synthetic wastewater could not be effectively removed by conventional coagulation and sedimentation. A similar condition was observed in the start-up period of the FPB operation. However, as a steady FPB was formed usually after about 10 hours' operation, the influent COD, ammonia nitrogen and total phosphorous could be quickly and effectively entrapped by the fluidized grown particle layer. Because these substances were not coagulable under normal conditions, adsorption might have performed an important role in their removal. Through an adsorption experiment using the grown pellets as adsorbent and glucose as adsorbate without dosing coagulants, it was found that the process followed well the Freundlich adsorption isotherm. The adsorption was also confirmed to be reversible by a washing experiment. A scanning electron microscopy coupled with energy dispersive spectroscopy analysis of the pellets before and after washing showed that the elements of carbon and phosphorous in the outer layer of the pellets were easily desorbed. The study results can provide an explanation, other than the coagulation mechanism, for the removal of soluble matter by the FPB.


Effects of recombinant porcine somatotropin on metabolic rate in growing pigs.
Effects of recombinant porcine somatotropin (rpST) on metabolic rate were studied in two trials with 24 crossbred barrows (Yorkshire x Landrace) in each. The barrows weighed about 80 kg (SE within trials 2.2 kg) at the start of the measurements and in each trial 12 pigs received 4 mg of rpST and 12 received a placebo. The diet contained 2.57 Mcal NE/kg and 20% CP (about 1% lysine). Animals were fed approximately 2.8 times maintenance (280 kcal ME.kg-.75.d-1). Heat production (gaseous exchange of CO2 and O2) and activity were measured continuously. Heat production associated with activity was calculated from the regression of heat production on activity. Animals treated with rpST exceeded controls in rate of gain by 252 g/d (P less than .001) and in metabolic rate by 14.5 kcal.kg-.75.d-1 (P less than .01). The rpST treatment increased rectal (+ .2 degrees C) and surface (+ .8 degrees C) temperatures. Activity-related heat production in treated pigs was increased, but this was only partly related to the increase in metabolic rate with rpST. The daily patterns of total and activity-related heat production were similar between pigs in both experimental treatments.


Novel Geometries for Stereotactic Localizers.
The N-localizer is generally utilized in a 3-panel or, more rarely, a 4-panel system for computing stereotactic positions. However, a stereotactic frame that incorporates a 2-panel (bipanel) N-localizer system with panels affixed to only the left and right sides of the frame offers several advantages: improved ergonomics to attach the panels, reduced claustrophobia for the patient, mitigation of posterior panel contact with imaging systems, and reduced complexity. A bipanel system that comprises two standard N-localizer panels yields only two three-dimensional (3D) coordinates, which are insufficient to solve for the stereotactic matrix without further information. While additional information to determine the stereotactic positions could include scalar distances from Digital Imaging and Communications in Medicine (DICOM) metadata or 3D regression across the imaging volume, both have risks related to noise and error propagation. Therefore, we sought to develop new stereotactic localizers that comprise only lateral fiducials (bipanel) that leave the front and back regions of the patient accessible but that contain enough information to solve for the stereotactic matrix using each image independently. Methods: To solve the stereotactic matrix, we assumed the need to compute three or more 3D points from a single image. Several localizer options were studied using Monte Carlo simulations to understand the effect of errors on the computed target location. The simulations included millions of possible combinations for computing the stereotactic matrix in the presence of random errors of 1mm magnitude. The matrix then transformed coordinates for a target that was placed 50mm anterior, 50mm posterior, 50mm lateral, or 50mm anterior and 50mm lateral to the centre of the image. Simulated cross-sectional axial images of the novel localizer systems were created and converted into DICOM images representing computed tomography (CT) images. Results: Three novel models include the M-localizer, F-localizer, and Z-localizer. For each of these localizer systems, optimized results were obtained using an overdetermined system of equations made possible by more than three diagonal bars. In each case, the diagonal bar position was computed using standard N-localizer mathematics. Additionally, the M-localizer allowed adding a computation using the Sturm-Pastyr method. Monte Carlo simulation demonstrated that the Z-localizer provided optimal results. The three proposed novel models meet our design objectives. Of the three, the Z-localizer produced the least propagation of error. The M-localizer was simpler and had slightly more error than the Z-localizer. The F-localizer produced more error than either the Z-localizer or M-localizer. Further study is needed to determine optimizations using these novel models.


STAT3 silencing in dendritic cells by siRNA polyplexes encapsulated in PLGA nanoparticles for the modulation of anticancer immune response.
In dendritic cells (DCs), the induction of signal transducer and activator of transcription 3 (STAT3) by tumor-derived factors (TDFs) renders DCs tolerogenic and suppresses their antitumor activity. Therefore, silencing STAT3 in DCs is beneficial for cancer immunotherapy. We have shown that STAT3 knockdown in B16 murine melanoma by siRNA polyplexes of polyethylenimine (PEI) or its stearic acid derivative (PEI-StA) induces B16 cell death in vitro and in vivo. Here, we investigated the physical encapsulation of siRNA/PEI and PEI-StA polyplexes in poly(d,l-lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) for STAT3 knockdown in DCs. PLGA NPs containing siRNA polyplexes of PEI (PLGA-P) and PEI-StA (PLGA-PS) had an average diameter of ~350 to 390 nm and a zeta potential of ∼-13 to -19 mV, respectively. The encapsulation efficiency (E.E.) of siRNA in PLGA-P and PLGA-PS was 26% and 43%, respectively. In both NP types, siRNA release followed a triphasic pattern, but it was faster in PLGA-PS. Our uptake study by fluorescence microscopy confirmed DC uptake and endosomal localization of both NP types. After exposure to B16.F10 conditioned medium, DCs showed high STAT3 and low CD86 expression indicating impaired function. STAT3 silencing by PLGA-P and PLGA-PS of STAT3 siRNA restored DC maturation and functionality as evidenced by the upregulation of CD86 expression, high secretion of TNF-α and significant allogenic T cell proliferation. Moreover, encapsulation in PLGA NPs significantly reduced PEI-associated toxicity on DCs. We propose this formulation as a strategy for targeted siRNA delivery to DCs. The potential of this approach is not limited to STAT3 downregulation in DCs but can be used to target the expression of other proteins as well. Moreover, it can be combined with other means for cancer immunotherapy like cancer vaccine strategies.


[Effects of exogenous nitric oxide on ascorbate-glutathione cycle in tomato seedlings roots under copper stress].
By using solution culture method, this paper studied the effects of exogenous nitric oxide (NO) on the antioxidants and antioxidases in the ascorbate-glutathione (AsA-GSH) cycle in tomato (Lycopersicon esculentum Mill. ) seedling roots under copper stress.Exogenous NO could affect the metabolic cycle of AsA-GSH in tomato roots under copper stress. Applying appropriate amount of exogenous NO increased the AsA and GSH contents, AsA/DHA and GSH/GSSG ratios, and decreased the DHA and GSSG contents in tomato roots under copper stress. With the addition of 100 micro mol L-1 of BSO, exogenous NO increased the AsA content, AsA/DHA ratio, and the AAO, MDHAR, and DHAR activities, and decreased the DHA, GSH, and GSSG contents and the APX and GR activities. When 250 micro mol L-1 of BSO was added, exogenous NO increased the contents of AsA, GSH, and GSSG, AsA/DHA ratio, and the activities of APX and GR, and decreased the DHA content and the AAO, DHAR and MDHAR activities. It was suggested that exogenous NO could affect the metabolic cycle of AsA-GSH in tomato roots under copper stress, and mitigate the damage of copper stress to tomato roots via regulating the AsA/DHA and GSH/GSSG ratios to alleviate oxidative stress.


Complete obstruction of the nasolacrimal system. Part II. Treatment with expandable metallic stents.
To evaluate the clinical usefulness of expandable metallic stents for maintenance of the dilated nasolacrimal sac and duct, double stents were implanted under fluoroscopic guidance in seven patients with severe epiphora after failed balloon dacryocystoplasty. Stents were constructed of 0.25-mm stainless steel wire in a cylindrical zigzag configuration of 10 bends. A single stent was 4 mm in diameter when fully expanded and 10 mm long. The stent was placed accurately in all but one patient, in whom a second stent was placed accurately after removal of the dislodged stent with a hemostat under endoscopic guidance. After stent placement, all patients demonstrated complete resolution of epiphora. During follow-up of 4-20 weeks, complete blockage of the stent occurred in one patient. This was treated with intrastent balloon dacryocystoplasty. No stents migrated. Although the long-term validity of this treatment has not been proved, expandable metallic stents seem to be of value in the treatment of complete obstruction of the lacrimal sac and nasolacrimal duct.


Differential trajectories of age-related changes in components of executive and memory processes.
Several studies have demonstrated age-related declines in general executive function and memory. In this study, we examined cross-sectional and longitudinal age effects in more specific cognitive processes that constitute executive function and memory. We postulated that, whereas some components of executive and memory functions would show age differences and longitudinal declines, other specific abilities would be maintained or even improve with repeated testing. In a sample of individuals ≥55 years old from the Baltimore Longitudinal Study of Aging, we found longitudinal declines in inhibition, manipulation, semantic retrieval, phonological retrieval, switching, and long-term memory over a maximum of 14 years follow-up. In contrast, abstraction, capacity, chunking, discrimination, and short-term memory were maintained or even improved longitudinally, probably due in part to repeated testing. Moreover, whereas several different abilities were correlated across participants' cross-sectional performance, longitudinal changes in performance showed more heterogeneous trajectories. Finally, compared with cross-sectional performance, longitudinal trajectories showed better distinction between participants with and those without later cognitive impairment. These results show that longitudinal cognitive aging of executive and memory functions is not a uniform process but a heterogeneous one and suggest that certain executive and memory functions remain stable despite age-related declines in other component processes.


Coping with an exulcerated breast carcinoma: an interpretative phenomenological study.
To explore how women living at home with a malignant fungating wound (MFW) cope with such wounds. To explore coping through the lived experiences of patients a methodological framework, using Heideggerian hermeneutic phenomenology and semi-structured interviews. Nine patients were interviewed from January until November 2009. The results are divided into two categories: 'living with a MFW' and 'feeling different'. These categories demonstrate how it is to live with the unpredictability, and uncontrollability of a MFW due to symptoms such as malodour, bleeding, exudate, pain and itching. The loss of control of the body boundary due to uncontrollable symptoms led to significant levels of distress and suffering for the patients. Different coping strategies were used to live with this wound. This study demonstrates how difficult it is to live and cope with a malignant fungating wound. Coping strategies, including going into isolation, or denying any issues, were used. When taking care of patients with MFWs, strategies need to integrate a palliative, holistic, empathic approach.


Investigating histidinylated highly branched poly(lysine) for siRNA delivery.
The temporary silencing of disease-associated genes utilising short interfering RNA (siRNA) is a potent and selective route for addressing a wide range of life limiting disorders. However, the few clinically approved siRNA therapies rely on lipid based formulations, which although potent, provide limited chemical space to tune the stability, efficacy and tissue selectivity. In this study, we investigated the role of molar mass and histidinylation for poly(lysine) based non-viral vectors, synthesised through a fully aqueous thermal condensation polymerisation. Formulation and in vitro studies revealed that higher molar mass derivatives yielded smaller polyplexes attributed to a greater affinity for siRNA at lower N/P ratios yielding greater transfection efficiency, albeit with some cytotoxicity. Histidinylation had a negligible effect on formulation size, yet imparted a moderate improvement in biocompatibility, but did not provide any meaningful improvement over silencing efficiency compared to non-histidinylated derivatives. This was attributed to a greater degree of cellular internalisation for non-histidinylated analogues, which was enhanced with the higher molar mass material.


Control of muscle regeneration in the Xenopus tadpole tail by Pax7.
The tail of the Xenopus tadpole will regenerate completely after transection. Much of the mass of the regenerate is composed of skeletal muscle, but there has been some uncertainty about the source of the new myofibres. Here, we show that the growing tail contains many muscle satellite cells. They are active in DNA replication, whereas the myonuclei are not. As in mammals, the satellite cells express pax7. We show that a domain-swapped construct, pax7EnR, can antagonize pax7 function. Transgenic tadpoles were prepared containing pax7EnR driven by a heat-inducible promoter. When induced, this reduces the proportion of satellite cells formed in the regenerate. A second amputation of the resulting tails yielded second regenerates containing notochord and spinal cord but little or no muscle. This shows that inhibition of pax7 action does not prevent differentiation of satellite cells to myofibres, but it does prevent their maintenance as a stem cell population.


Molecular epidemiology and evolution of emerging infectious diseases.
Molecular epidemiology is an emerging science. The development of new and rapid protocols to isolate and identify pathogens, coupled with the sophisticated phylogenetic analysis of their gene sequences, is providing a new and fascinating insight into the biology, origin and spread of infectious diseases. In this essay, I describe some of the ways in which the techniques of modern molecular biology and evolution have equipped us to face the challenge of these new infections.


[Serological assay for diagnosis of verotoxin-producing Escherichia coli (VTEC) infection in the patients with diarrhea].
A total of 239 serum samples from 136 persons were used for bacterial agglutination assay (BA) against predominant three O-antigens of VTEC. All VTEC isolates from stools of 30 patients were only O157:H7 serotype (these patients are called group I). The levels of positive BA antibody titers (over 1:160) to O157-antigen were recognized in each patients as follows. The VTEC isolated patients with HUS or without HUS in group I were all of 13 (100%) and 14 (82.4%) in 17 patients, respectively. And 21 (65.6%) patients of group II (HUS patients with stool negative cultures, or stool cultures were not performed in 32 patients), and 6 (15.0%) patients of group III (family members of group I and II; 40 persons), were also recognized. In group IV (patients with diarrhea due to other pathogen than VTEC; 11 patients), and V (clinically healthy persons; 23 persons), none were recognized as positive BA antibody titers. All patients in the group II except one who had a positive BA antibody titer to O111, were not recognized to O111 and O26. A few VTEC-positive patients without gastrointestinal syndrome did not have significant agglutinating titers to O157-antigen on the days after VTEC isolation. However, almost all patients with diarrhea due to VTEC and HUS, and with VTEC but no HUS, had a level of positive BA antibody titer on the 5 day after onset of diarrhea. These results suggest that this serological assay is a very simple and useful tool for diagnosis of VTEC infection when VTEC are not detected by culture method due to antimicrobial treatment, or due to the lapse of many days after onset of diarrhea.


Synovial chondromatosis originating from the synovium of the anterior cruciate ligament: a case report.
A case of synovial chondromatosis originating from the synovium of the anterior cruciate ligament (ACL) resulting in a mechanical block to knee extension is reported. A 36-year-old man complained of a restricted range of left-knee motion and pain when walking. Plain roentgenograms showed normal appearance, however, magnetic resonance imaging showed intensity changes in the ACL. Arthroscopically, numerous small free bodies were observed. Proliferation of synovium and cartilaginous tissues were identified around the ACL. There were no significant findings in the synovium except around the ACL. The synovium around the ACL was resected and free bodies were washed out. This is the first report of synovial chondromatosis originating from the synovium of the ACL.


Are diagnostic lumbar medial branch blocks valid? Results of 2-year follow-up.
The precise cause of low back pain based on clinical history, physical examination, radiological imaging, and electrophysiological testing can be identified in only 15% of patients in the absence of disc herniation and neurological deficit. The prevalence of chronic lumbar zygapophysial (facet) joint pain ranges from 15% to 45% utilizing comparative local anesthetic blocks in controlled settings in accordance with the criteria established by the International Association for the Study of Pain. Currently, facet joint injection procedures are considered as the gold standard in the diagnosis of facet joint pain. Facet blocks have been criticized as lacking diagnostic validity, along with other tests, including discography, neurophysiologic tests, stress radiographs, x-ray studies, bone scintigraphy, thermography and diagnostic ultrasound. However, these assumptions have been based on biased evaluations without consideration of the criteria of the International Association for the Study of Pain, as well as the nature of controlled diagnostic blocks. Utilizing the criteria established by the Agency for Healthcare Research and Quality (AHRQ) for Systems to Rate the Strength of Scientific Evidence, it was shown that the validity, specificity, and sensitivity of facet joint nerve blocks are considered strong in the diagnosis of facet joint pain. The accuracy of a diagnostic test is best determined by comparing it to an appropriate reference standard, such as biopsy, surgery, autopsy or long-term follow-up. Since we are unable to apply reference standards of biopsy, surgery, or autopsy, and pain relief has been argued as an inconsistent feature, long-term follow-up has been considered as the best indicator. This study was undertaken to evaluate stability of the diagnosis of lumbar facet joint pain following comparative local anesthetic blocks at a follow-up after 2 years. The results showed that 85% of the patients available for follow-up withstood the diagnosis of facet joint pain at the end of 2 years, whereas this proportion decreased to 75%, if all the patients in the study were included in the analysis.


[The effect of vagotomy on carcinogenesis in the glandular stomach of rats, induced by N-methyl-N'-nitro-N-nitrosoguanidine].
The effect of hydrochlorhydria caused by vagotomy on carcinogenesis in the glandular stomach of male rats was studied. Group A (35 rats): After N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) 80 mg/l solution was orally administered during the first 15 weeks of life, vagotomy was performed. Group B (35 rats): After the oral administration of MNNG 80 mg/l solution during the first 15 weeks of life, laparotomy was done. Group C (10 rats): As the control, vagotomy was undertaken at the 15th week of life. Group D (10 rats): As the control, laparotomy was done at the same time. At the 52nd week, all surviving rats were autopsied, and gastrin cell counts and body weight were ascertained. The incidence of adenocarcinoma was 62% in Group A, 32% in Group B (p less than 0.05) and nil in Group C and D. These results strongly support the view that the hypochlorhydria plays the role of a promoting factor in producing gastric carcinoma.


Evidence for segment-nonspecific packaging of the influenza a virus genome.
The influenza A virus genome is composed of eight negative-sense RNA segments (called vRNAs), all of which must be packaged to produce an infectious virion. It is not clear whether individual vRNAs are packaged specifically or at random, however, and the total vRNA capacity of the virion is unknown. We have created modified forms of the viral nucleoprotein (NP), neuraminidase (NA), and nonstructural (NS) vRNAs that encode green or yellow fluorescent proteins and studied the efficiency with which these are packaged by using a plasmid-based influenza A virus assembly system. Packaging was assessed precisely and quantitatively by scoring transduction of the fluorescent markers in a single-round infectivity assay with a flow cytometer. We found that, under conditions in which virions are limiting, pairs of alternatively tagged vRNAs compete for packaging but do so in a nonspecific manner. Reporters representing different vRNAs were not packaged additively, as would be expected under specific packaging, but instead appeared to compete for a common niche in the virion. Moreover, 3 to 5% of transduction-competent viruses were found to incorporate two alternative reporters, regardless of whether those reporters represented the same or different vRNAs - a finding compatible with random, but not with specific, packaging. Probabilistic estimates suggest that in order to achieve this level of dual transduction by chance alone, each influenza A virus virion must package an average of 9 to 11 vRNAs.


Intestinal cryptosporidiosis in turkeys in Iran.
Naturally occurring cryptosporidiosis is reported in turkey poults suffering from diarrhoea and unthriftness in Iran. Histological and ultrastructural studies revealed high number of Cryptosporidium developmental stages mainly located in the mid and terminal portions of small intestine of the poults. Other portions of the intestinal tract were less frequently infected. Oocyst shedding was detected only in 29% of the histologically positive birds. Based on host species, clinical signs, pathology and tissue location of the parasites, Cryptosporidium meleagridis was most likely responsible for these infections. This parasite was also reported recently in a child with diarrhoea in Iran.


Nanoscale cues regulate the structure and function of macroscopic cardiac tissue constructs.
Heart tissue possesses complex structural organization on multiple scales, from macro- to nano-, but nanoscale control of cardiac function has not been extensively analyzed. Inspired by ultrastructural analysis of the native tissue, we constructed a scalable, nanotopographically controlled model of myocardium mimicking the in vivo ventricular organization. Guided by nanoscale mechanical cues provided by the underlying hydrogel, the tissue constructs displayed anisotropic action potential propagation and contractility characteristic of the native tissue. Surprisingly, cell geometry, action potential conduction velocity, and the expression of a cell-cell coupling protein were exquisitely sensitive to differences in the substratum nanoscale features of the surrounding extracellular matrix. We propose that controlling cell-material interactions on the nanoscale can stipulate structure and function on the tissue level and yield novel insights into in vivo tissue physiology, while providing materials for tissue repair.


[How to prevent and treat glaucoma surgical failures?].
Although many improvements have been made to filtering surgery in glaucoma, some post operative complications do occur quite often. Patients need careful and frequent follow-up to assess and treat the early complications that may occur after glaucoma surgery. Over the medium and long term, insufficient verification of the intra ocular pressure signals the failure of the initial surgery. In this article, we review the diagnostic tools useful for detecting the early and late failures of glaucoma surgery and we provide recommendations for their treatment.


[Ophthalmomyiasis caused by Oestrus ovis in a 3 year old boy].
Oestrus ovis larvae are obligatory parasites of the nasal and sinus cavities of sheep and goats. Oestrosis in small ruminants is prevalent in Mediterranean and tropical countries. Occasionally, humans are affected by ocular myiasis (ophthalmomyiasis) and more rarely, nasal myiasis. A case of human ophthalmomyiasis caused by first instar larvae of Oestrus ovis in a 3 year old boy was observed in Gran Canaria. Acute catarrhal conjunctivitis developed within few hours. During the inspection of the eye several larvae could be observed, moving quickly over the cornea to the conjunctival sac. Rinsing and mechanical removal of the larvae lead to an uncomplicated recovery.


Cluster migraine--an unrecognized common entity.
A review is presented of a distinctive type of vascular headache and its stereotyped features are emphasized, viz. severe non-throbbing character, unilateral and usually peri-ocular localization, frequent nocturnal occurrence and tendency to occur in brief episodes in a clustered pattern over weeks or months, with headache-free periods lasting months or years.Ten illustrative case histories are given and a plea is made for the early clinical recognition of this entity so that patients will not be submitted to unnecessary and potentially hazardous investigative procedures. While the common analgesics and narcotics are usually ineffective, ergotamine derivatives, methysergide and a new compound, BC-105, are highly effective forms of therapy.


Archaeal tyrosine recombinases.
The integration of mobile genetic elements into their host chromosome influences the immediate fate of cellular organisms and gradually shapes their evolution. Site-specific recombinases catalyzing this integration have been extensively characterized both in bacteria and eukarya. More recently, a number of reports provided the in-depth characterization of archaeal tyrosine recombinases and highlighted new particular features not observed in the other two domains. In addition to being active in extreme environments, archaeal integrases catalyze reactions beyond site-specific recombination. Some of these integrases can catalyze low-sequence specificity recombination reactions with the same outcome as homologous recombination events generating deep rearrangements of their host genome. A large proportion of archaeal integrases are termed suicidal due to the presence of a specific recombination target within their own gene. The paradoxical maintenance of integrases that disrupt their gene upon integration implies novel mechanisms for their evolution. In this review, we assess the diversity of the archaeal tyrosine recombinases using a phylogenomic analysis based on an exhaustive similarity network. We outline the biochemical, ecological and evolutionary properties of these enzymes in the context of the families we identified and emphasize similarities and differences between archaeal recombinases and their bacterial and eukaryal counterparts.


Fifteen-Year Trends in the Prevalence of Diabetes among Hospitalized HIV-Infected Patients in Spain (1997-2012).
To describe trends in the prevalence of diabetes among hospitalized HIV-infected patients between 1997 and 2012 in Spain and compare them with those of age- and sex-matched non-HIV-infected patients. The study was based on Spanish national hospital discharge data. We performed a retrospective study for the period 1997-2012. HIV infection (HIV-infected versus non-HIV-infected [control group])and calendar period in relation to widespread use of combination antiretroviral therapy (cART) (1997-1999; 2000-2003; 2004-2007 and 2008-2012), were the exposure variables The outcome variables were diagnosis of diabetes and in-hospital mortality (IHM). From 1997 to 2012, we identified 91,752 cases of diabetes: 15,398 in the HIV-infected group (403,277 hospital admissions) and 76,354 in the non-HIV-infected group (1,503,467 hospital admissions). Overall, HIV-infected patients had lower prevalence values for diabetes than non-HIV-infected patients throughout the follow-up (3.8% vs. 5.1%; p<0.001). The prevalence of diabetes increased 1.56-fold among non-HIV-infected patients and 4.2-fold among HIV-infected patients. The prevalence of diabetes in females was almost twice as high in HIV-infected patients as in non-HIV-infected patients during the last study period (4.72% vs. 2.88%; p<0.001). Diabetes showed a protective effect against IHM throughout the study period (aOR = 0.70; 95%CI, 0.65-0.75). During the cART era, the prevalence of diabetes has increased sharply among HIV-infected hospitalized patients compared with matched non-HIV-infected subjects. The prevalence of diabetes is rising very fast among HIV-infected women. Diabetes has a protective effect on IHM among HIV-infected patients. Nevertheless, our study has several limitations. No information is available in the database used on important sociodemographic characteristics and relevant clinical variables including duration of the HIV infection, treatments used, drug resistance, treatment adherence or CD4 count, among others. Also, it is possible that increase of diabetes prevalence could reflect the improvement in recording habits.


Treatment of dyslipidaemia in high-risk patients: too little, too late.
Evidence that lowering low-density cholesterol (LDL-C) reduces coronary events and mortality is now overwhelming and is reflected in treatment guidelines from around the world. The Joint European Guidelines recommend an LDL-C goal of <3.0 mmol/l in high-risk subjects. The National Cholesterol Education Program (NCEP) Adult Treatment Panel (ATP)-III guidelines suggest an even more aggressive approach in high-risk individuals, with a recommended LDL-C goal of <2.6 mmol/l. Large numbers of high-risk patients are still not achieving the more conservative goals recommended in the Joint European Guidelines, let alone the more aggressive LDL-C target recommended in the new NCEPATP-III guidelines. The recognition in the NCEP ATP-III guidelines that a high-density lipoprotein cholesterol (HDL-C) level <1.0 mmol/l represents an important risk factor highlights the emergence of HDL-C as a key player in the genesis of coronary heart disease (CHD) and as a potential target for therapy. This may be especially important in people with insulin resistance with or without type 2 diabetes. There is evidence from the Helsinki Heart Study and the more recent Veterans Affairs HDL Intervention Trial (VA-HIT), both of which used gemfibrozil as the active agent, that the observed reduction in coronary events was correlated with the magnitude of the increase in HDL-C. The challenge for future management of high-risk individuals will be not only to reduce the level of LDL-C to below 2.6 mmol/l but also to increase HDL-C to levels above 1.0 mmol/l.


Invasiveness-triggered state transition in malignant melanoma cells.
Cancer cells are considered to have high morphological heterogeneity in human melanoma tissue. Here, we report that epithelial cancer cells are dominant in different development stages of human melanoma tissues. The cellular and molecular mechanisms that maintain melanoma cells in the epithelial state are further investigated in the A2058 cell line. We find that micropore (8 µm) transwell invasion, but not superficial migration in the scratch assay, can induce remarkable morphological changes between epithelial and mesenchymal melanoma cells within 4 days. The morphological switch is associated with dynamic changes of epithelial-mesenchymal transition (EMT) hallmarks E-cadherin and vimentin. Further immunoflurencent staining and co-immunoprecipitation assay showed the uncoupling of the M3 muscarinic acetylcholine receptor (mAChR) and the p75 neurotrophin receptor (p75NTR) in epithelial melanoma cells. Specific knockdown of M3 mAChR by small interfering RNA (siRNA) significantly abrogates the transition of spindle-shaped mesenchymal cells to epithelial cells. Collectively, we report a cellular model of invasiveness-triggered state transition (ITST) in which melanoma cell invasion can induce morphological changes between epithelial and mesenchymal cells. ITST is one of the biological basis for maintaining metastatic melanoma cells in the epithelial state. Furthermore, M3 mAChR receptor-mediated ITST provides a novel therapeutic strategy to inhibit the development of malignant melanoma.


[Evaluation of operating theatre for the risk prevention].
In this study we analyse factors that predispose to risk and we value the importance of quality and reliability into operating theatre. Patient safety result from ability to plan and manage organizations able to reduce probability of mistakes (Prevention) and to recover and contain their consequences (Protection). The principal motives of mistakes are: - deficiency of sharing procedures between different professional figures for risk prevention; - deficiency of an effective integration between professional figures in operating theatre; - deficiency of charitable precise run to guarantee the continuity of interventions on patient. A risk management program have to take care on this sentinel events and to set up a survey-information system to characterize risks and correction strategies. To prevent patient change, wrong side identification, not working devices or deficiency of surgery materials, it would be useful for operating theatre figures and for anesthetists to work out all together a procedure for admission in operating theatre and a pre-operating check list. The best way to carry out a safety and quality attendance based on standardized procedures and protocols, is to set up risk management firm units. It is necessary to activate and to adjust survey system and effective management training.


Inorganic contaminants in Canadian First Nation community water systems.
While previous Canadian studies have examined microbiological water quality in First Nations, there is little published information on inorganic contaminants. In Atlantic Canada, the lead, manganese, and arsenic content of First Nations' drinking water has been measured for more than a decade, but the data have not been analyzed comprehensively. These contaminants are linked with health problems, and high levels in drinking water are a cause for concern. We examined 12 years of data from 47 First Nation community water systems to identify systems experiencing difficulties meeting sampling frequency or regulatory guidelines. While most contaminant concentrations were below guideline values, we identified elevated concentrations and issues with sampling frequency. No system met both sampling frequency requirements - a minimum of one sample per year per analyte - and regulatory guidelines. Exceedance rates for lead, manganese, and arsenic were high in some systems. Moreover, current sampling procedures for lead specify that taps be flushed prior to sampling, which is known to underestimate lead exposure. We find that a switch to random daytime sampling would at least sometimes yield higher estimates of lead at the tap. Our analysis demonstrates the need for increased monitoring and updated sampling procedures to better characterize inorganic contaminant occurrence in First Nations.


Successful Outcome of Inhalation Injury, Active SARS-CoV-2 Infection, and Concomitant Pneumonia in a Patient With 27% Full-Thickness Burn: A Case Report.
Burn injuries are a major cause of morbidity and mortality. Next to the inhalation injury, TBSA and age are strong predictors of mortality in burn victims. The novel coronavirus disease 2019 (COVID-19) pandemic is associated with a fatality rate of around 3.5%. We present a case of burn victim with full-thickness burn to face, scalp, both upper extremities (27% of TBSA), inhalation injury, and active severe acute respiratory syndrome coronavirus 2 infection with concomitant pneumonia. The inhalation injury in COVID-19 positive patient was severe. A bronchoscopy revealed a diffuse erythema of the trachea and both main bronchi, the whole bronchial tree up to the distal segments was covered with carbonaceous material which could not be removed. We decided to treat the inhalation injury according to the guidelines for burns and acute respiratory distress syndrome. Accordingly, the patient did not receive any antiviral drugs or corticosteroids. The reconstruction of a full-thickness scalp defect after burn presents a challenge in large size defects and in patients with comorbidities. Double layer Integra Dermal Regeneration Template (Integra LifeSciences, Plainsboro, New Jersey) was the reconstruction method of choice. The take of dermal template and split-thickness skin graft was 100% and good scalp contour was achieved. To our knowledge this is the first case report presenting a successful treatment outcome in a burn victim with inhalation injury, active severe acute respiratory syndrome coronavirus 2 infection, and concomitant pneumonia with full-thickness burn of 27% of TBSA.


Clastogenic effect of bracken fern (Pteridium aquilinum v. arachnoideum) diet in peripheral lymphocytes of human consumers: preliminary data.
Ingestion of bracken fern (Pteridium aquilinum v. arachnoideum) is associated with digestive tract cancer in different regions of Japan, Venezuela and Brazil. In view of reports that dietary bracken fern causes chromosomal instability in cattle, the clastogenic effect of bracken fern was investigated, in a preliminary study, in peripheral lymphocytes obtained from habitual consumers and a control group of non-consumers, which were carefully investigated about cancer history or family cancer history, negative in both cases, using protocols comparable to those previously described in studies in cattle raised on bracken pastures. Cytogenetic analysis showed significant increased levels of chromosomal abnormalities, such as chromatid breaks, in cultured peripheral lymphocytes of the consumer group. There was no correlation with subjects, gender, smoking habits or alcohol consumption, and the only correlation was with prolonged exposure to dietary bracken.


Does aging stop?
Human mortality data show stabilization in mortality rates at very late ages. But human mortality data are difficult to interpret because they are affected by changing medical practices and other historically variable causes of death. However, in the 1990s, data from a variety of labs showed that the mortality rates of medflies, fruit flies, wasps, yeasts, and nematodes also stabilize at very late ages. These reproducible "mortality-rate plateaus" forced biologists to develop theories for their existence. There are two main theories of this kind. "Lifelong heterogeneity" theories suppose that highly robust subcohorts are more abundant at later ages because less robust subcohorts have mostly died off. On this type of theory, aging does not stop; aging continues inexorably in all individuals. In contrast, in evolutionary theories for mortality-rate plateaus, based on the eventual plateaus in Hamilton's Forces of Natural Selection at late ages, aging does indeed stop. A variety of experiments have cast doubt on lifelong heterogeneity theories as explanations of mortality-rate plateaus. A few experiments have corroborated the Hamiltonian theory. This has the important corollary that it appears to be possible for aging to stop, at sufficiently late ages, at least among some populations. The implications of this result for aging research are profound. Most importantly, it suggests the possibility that the physiology of adults undergoing aging may be substantially different from the physiology of life after aging.


Antibacterial activity of WY-49605 compared with those of six other oral agents and selection of disk content for disk diffusion susceptibility testing.
The in vitro antimicrobial activity of an oral penem, WY-49605, was compared with those of six other oral antimicrobial agents against 598 bacterial isolates representing 51 different species. WY-49605 exhibited good activity against most gram-positive bacteria and members of the family Enterobacteriaceae. It had little activity against nonfermenting gram-negative bacilli, Enterobacter spp., Serratia spp., enterococci, Staphylococcus haemolyticus, and methicillin-resistant Staphylococcus aureus. Its activity was unaffected by the beta-lactamases of Neisseria gonorrhoeae, Haemophilus influenzae, and staphylococci. Disk diffusion susceptibility tests were performed with 5-, 10-, 15-, and 30-micrograms WY-49605 disks. The 5-micrograms disk is recommended, with tentative breakpoints of > or = 16 mm for susceptibility (MIC, < or = 2.0 microgram/ml) and < or = 12 mm for resistance (MIC, > or = 8.0 micrograms/ml).


Characterization of lipopolysaccharide heterogeneity in Salmonella enteritidis by an improved gel electrophoresis method.
Salmonella enteritidis field isolates of different phage types and pathogenicities were assessed for changes in lipopolysaccharide (LPS) structure, using an improved method of polyacrylamide gel electrophoresis (PAGE) that revealed the same degree of structural detail as mass spectroscopy. The method allowed characterization of an LPS chemotype that may be associated, regardless of phage type, with increased virulence of S. enteritidis. The virulent variant SE6-E21, which efficiently contaminates eggs and yields high numbers of organisms from chick spleens, had an O-antigen/core ratio of 2.8, as determined from gels by densitometry, and 1.67 micrograms of mannose per microgram of 2-keto-3-deoxy-octulosonic acid (KDO), while the avirulent variant SE6-E5 had O-antigen/core ratios of 1.2 and 1.00. The association between O antigen and virulence was also seen on analysis of five new field isolates. One of the new field isolates generated a mixed population of smooth and semismooth variants in agreement with its mixed virulence in chicks. When LPS was purified from large-volume cultures, only the most virulent isolate yielded high amounts of O antigen (1.6 micrograms of mannose per microgram of KDO), while the other isolates had ratios characteristic of semismooth variants (< or = 1.0 microgram of mannose per microgram of KDO), including the isolate of mixed virulence. These results indicate that the improved PAGE method might provide a rapid, sensitive, in vitro assessment of field isolate virulence prior to the performance of definitive infectivity trials.


Comparison of the Anesthetic Effects of Isoflurane and Sevoflurane Inhalation on the Cardiovascular System of the Common Buzzard (Buteo buteo).
The aim of this study was to compare the effects of isoflurane (ISO) and sevoflurane (SEVO) inhalation anesthesia on the cardiovascular system of the common buzzard (Buteo buteo). Fourteen common buzzards of unknown sex and age, weighing 700-900 g each, were used for the study. The buzzards were randomly divided into an ISO group and a SEVO group, each comprising 7 buzzards. The buzzards were then anesthetized by masked induction with either ISO or SEVO. After induction, all buzzards were intubated with a 2.5-mm-diameter uncuffed endotracheal tube. Anesthesia was maintained with 3% ISO or 4% SEVO in a 3 L/min flow of 100% oxygen. Before induction, immediately after induction, and at intervals of 5, 10, 15, 20, 25, and 30 min after the patients were induced, the heart rate, respiratory rate, and body temperature of the buzzards were recorded. In addition, before and during anesthesia, serum electrolytes (sodium, ionized calcium, and potassium), blood gas measurements (arterial pH, arterial partial pressure of carbon dioxide, arterial partial pressure of oxygen, bicarbonate concentration, and oxyhemoglobin saturation), and hematocrit values were evaluated in the blood samples taken from both the ISO and SEVO groups. A statistically significant decrease in body temperature was observed in both groups. A significant decrease in heart rate was observed in the ISO group. The increases in partial pressure of carbon dioxide and bicarbonate concentration were statistically significant in both groups. The effects of ISO and SEVO anesthesia on the cardiovascular and respiratory system in the buzzards were similar based on the results of this study.


Dealing with uncertainty: statistics for an aging population.
"Uncertainty in statistics and demographic projections for aging and other policy purposes comes from four sources: differences in definitions, sampling error, nonsampling error, and scientific uncertainty. Some of these uncertainties can be reduced by proper planning and coordination, but most often decisions have to be made in the face of some remaining uncertainty. Although decision makers have a tendency to ignore uncertainty, doing so does not lead to good policy-making. Techniques for estimating and reporting on uncertainty include sampling theory, assessment of experts' subjective distributions, sensitivity analysis, and multiple independent estimates." The primary geographical focus is on the United States.


Cooperative metal binding and helical folding in model peptides of treble-clef zinc fingers.
Two peptides, L(TC) and L(TC)(T) have been synthesised to model the treble-clef zinc fingers encountered in many Zn(Cys)(4)-site-containing proteins. Both are cyclic peptides with a linear tail grafted on a glutamate side chain of the cycle. They differ by the length of this tail, which lacks five amino acids in L(TC)(T) compared to L(TC). Both peptides bind Zn(2+) and Co(2+) in 1:1 metal/peptide ratio and the structure of these complexes have been characterised by NMR, UV/Vis and CD spectroscopy. Both peptides fold the same way around the metal ion and they fully reproduce the classical fold of treble-clef zinc fingers and display an extended hydrogen-bond network around the coordinating sulfur atoms. The structures of the ML(TC) complexes reveal that the linear tail forms a short two-turn alpha-helix, present in the metallated form only. The formation of this helix constitutes a rare example of metal-induced folding. The second turn of this helix is composed of the five amino acids that are absent in L(TC)(T). The study of the pH-dependence of the Zn(2+) binding constants shows that the metal ion is bound by four cysteinates above pH 5.2 and the binding constants are the highest reported so far. Interestingly, the binding constant of Zn x L(TC) is about tenfold higher than that of Zn x L(TC)(T). This difference clearly indicates that the helix, present in Zn x L(TC) only, stabilises the Zn(2+) complex by about 1.2 kcal mol(-1). The origin of this stabilisation is ascribed to an electrostatic interaction between the [ZnS(4)](2-) centre and the helix. This reveals a cooperative effect: zinc binding allows the folding of the tail into a helix which, in turn, strengthens the zinc complex.


Evidence-Informed Update of Argentina's Health Benefit Package: Application of a Rapid Review Methodology.
Argentina has a fragmented healthcare system with social security covering almost two thirds of the population. Its benefit package-called compulsory medical program (PMO; by its Spanish acronym Programa Médico Obligatorio)-has not been formally and widely updated since 2005. However, laws, clinical practice guidelines (CPGs), and a high-cost technology reimbursement fund complement it. Our objective was to comprehensively review such a PMO and propose an update considering the corresponding complementary sources. We followed four steps: (i) identification of health technologies from the current PMO and complementary sources, (ii) prioritization, (iii) assessment through rapid health technology assessment (HTA), and (iv) appraisal and recommendations. We evaluated three value domains: quality of evidence, net benefit, and economics, which were summarized in a five-category recommendation traffic-light scale ranging from a strong recommendation in favor of inclusion to a strong recommendation for exclusion. Eight hundred fifty technologies were identified; 164 of those, considered as high priority, were assessed through rapid HTAs. Those technologies mentioned in laws and CPGs were mostly outpatient essential medicines, whereas those from the reimbursement system were mostly high-cost drugs; of these 101 technologies, 50 percent were recommended to be kept in the PMO. The other 63 (identified by the Superintendence of Health Services, technology producers, and patients) were mostly medical procedures and high-cost drugs; only 25 percent of those resulted in a favorable recommendation. A methodology based on four clearly identified steps was used to carry out a comprehensive review of an outdated and fragmented benefit package. The use of rapid HTAs and a traffic-light recommendation framework facilitated the deliberative evidence-based update.


Intralesional infusion of lymphokine-activated killer (LAK) cells and recombinant interleukin-2 (rIL-2) for the treatment of patients with malignant brain tumor.
Twenty patients with supratentorial, intracerebral lesions defined by computed tomographic scan or magnetic resonance imaging were treated by surgery and adoptive immunotherapy with lymphokine-activated killer (LAK) cells and recombinant Interleukin-2 (rIL-2, Cetus). Seventeen patients had glioblastoma, two had high-grade oligodendroglioma, and one patient had two metastatic sarcoma lesions. LAK cells were produced from blood mononuclear cells (MNC) obtained by 2 to 3 leukapheresis procedures and cultured (2.5 x 10(6) MNC/ml) 3 to 5 days with 1000 units rIL-2/ml. Although LAK cells could be produced from MNC of all patients, those taking steroids or with a low Karnofsky functional status generated, on average, suboptimal LAK cell activity. Age, sex, and serum anticonvulsant levels do not seem to influence a patient's ability to produce LAK cells in vitro. For therapy, cultured MNC (1-15 x 10(9] containing LAK cells were suspended in saline containing 10(6) units rIL-2 and injected into tissue surrounding the tumor cavity during craniotomy. For 3 days after their operations, patients received 10(6) units rIL-2 into the tumor cavity through an Ommaya reservoir. The treatment protocol was tolerated well by all patients, although they all experienced some degree of headache, fever, or lethargy that cleared within a few days of the last rIL-2 injection. When computed tomographic (CT) scans were obtained soon after treatment, areas of low density suggested a greater-than-normal extent of edema around the operative site. At the present time, CT scans indicate that the tumors of seven patients have recurred with an average disease-free interval of 25 +/- 6 weeks.(ABSTRACT TRUNCATED AT 250 WORDS)


From gut microflora imbalance to mycobacteria infection: is there a relationship with chronic intestinal inflammatory diseases?
The gut of a healthy adult harbours a myriad of different microbial species. It is estimated that approximately 10 14 are present in total bacterial colony forming units (CFU). Each colony colonizes a specific intestinal tract. In healthy adult, the main control of intestinal bacterial colonization occurs through gastric acidity but also other factors can influence the intestinal microenvironment such as pH, temperature, competition among different bacterial strains, peristalsis, drugs, radiotherapy and much more. Impaired microbial homeostasis leads to an alteration of the permeability of tissue, together with the activation of the intestinal immune system MALT (mucosal associated lymphoid tissue). In this regard we discuss the increasing experimental evidences of the role of commensal microbiota in the activation of specific intestinal immunocompetent cells. The aforementioned micro-environmental changes provide the substrate for the etiopathogenetic outbreak of numerous pathologies of gastro-intestinal tract, such as intestinal chronic inflammation (Crohn's disease and Ulcerative Colitis), together with a miscellany of extra intestinal disorders. This article is an overview of the latest scientific findings about the close causal relationship between intestinal microbial flora and inflammatory bowel diseases or other extra-intestinal diseases; it is also mentioned the possible relationship between mycobacteria and Chron's disease. Finally we analyse the beneficial role of probiotics.


Scanning Electrochemical Microscopy with Forced Convection Introduced by High-Precision Stirring.
In the present report the well-known advantages of hydrodynamic mass transport in electrochemical systems are used in combination with scanning electrochemical microscopy (SECM). The hydrodynamic SECM system integrates a high-precision stirring device into the experimental setup. The well-defined stirring of the SECM electrolyte results in steady state diffusion layer characteristics in the vicinity of large substrate electrodes operated in chronoamperometric measuring mode. For a range of rotation frequencies of a rotating cylinder the thickness and the stability of the diffusion layer was studied by hydrodynamic SECM in the substrate generation/tip collection (SG/TC) as well as in the competition mode using ferrocene methanol as redox mediator. Different UME probe dimensions ranging from Pt diameters of 20 μm down to 0.6 μm were used. The smallest probe with dPt = 0.6 μm was found most suitable for these studies due to the almost convection-independent amperometric response associated with subμm electrodes. Additionally, preliminary studies of hydrodynamic SECM imaging of a 2 mm Pt disk electrode surface in the SG/TC mode based on in situ produced hydrogen as a mediator are presented. Comparative images measured in the conventional positive feedback mode in quiescent solution show that hydrodynamic SECM offers attractive complementary information.


Examining the motivations and future career aspirations of Qatari pharmacy students and alumni: A case study.
The Global Pharmacy Workforce report published by the International Pharmaceutical Federation indicated pharmacy workforce shortages and called forstrengthening workforce planning. The low percentage of Qatari students in the College of Pharmacy at Qatar University could negatively impact the fulfilment of the Qatar National Vision and the United Nations' Goals. The aim of this research is to examine the motivations of Qatari pharmacy students and alumni to study pharmacy, and to understand their perceptions of the profession and career aspirations. A qualitative case study research was conducted. Data were collected by focus groups and semi-structured interviews, and were analysed thematically. Research findings suggested that increasing the national workforce was the main motive for students to study pharmacy. The participants' perceptions of the profession varied from pharmacy being a drug sales profession to it being a healthcare profession. Participants' career aspirations included pursuing graduate studies, working in hospitals, and working in academia. The participants were generally satisfied with studying pharmacy, which encouraged them to recommend the major to others. However, the participants were challenged by the lack of societal recognition of pharmacy and by misperceptions about Qatari students, which motivated them to further demonstrate their competencies. This study will facilitate the understanding of factors that motivate and demotivate Qatari students from joining the pharmacy major and their career aspirations, which will ultimately enhance the development of recruitment strategies geared towards attracting a larger number of qualified and highly motivated Qataris into the pharmacy profession.


A facial reconstruction method based on new mesh deformation techniques.
This article presents a new numerical method for facial reconstruction. The problem is the following: given a dry skull, reconstruct a virtual face that would help in the identification of the subject. The approach combines classical features as the use of a skulls/faces database and more original aspects: (1) an original shape matching method is used to link the unknown skull to the database templates; (2) the final face is seen as an elastic 3D mask that is deformed and adapted onto the unknown skull. In this method, the skull is considered as a whole surface and not restricted to some anatomical landmarks, allowing a dense description of the skull/face relationship. Also, the approach is fully automated. Various results are presented to show its efficiency.


Total joint arthroplasty: practice variation of physiotherapy across the continuum of care in Alberta.
Comprehensive and timely rehabilitation for total joint arthroplasty (TJA) is needed to maximize recovery from this elective surgical procedure for hip and knee arthritis. Administrative data do not capture the variation of treatment for rehabilitation across the continuum of care for TJA, so we conducted a survey for physiotherapists to report practice for TJA across the continuum of care. The primary objective was to describe the reported practice of physiotherapy for TJA across the continuum of care within the context of a provincial TJA clinical pathway and highlight possible gaps in care. A cross-sectional on-line survey was accessible to licensed physiotherapists in Alberta, Canada for 11 weeks. Physiotherapists who treated at least five patients with TJA annually were asked to complete the survey. The survey consisted of 58 questions grouped into pre-operative, acute care and post-acute rehabilitation. Variation of practice was described in terms of number, duration and type of visits along with goals of care and program delivery methods. Of the 80 respondents, 26 (33 %) stated they worked in small centres or rural settings in Alberta with the remaining respondents working in two large urban sites. The primary treatment goal differed for each phase across the continuum of care in that pre-operative phase was directed at improving muscle strength, functional activities were commonly reported for acute care, and post-acute phase was directed at improving joint range-of-motion. Proportionally, more physiotherapists from rural areas treated patients in out-patient hospital departments (59 %), whereas a higher proportion in urban physiotherapists saw patients in private clinics (48 %). Across the continuum of care, treatment was primarily delivered on an individual basis rather than in a group format. Variation of practice reported with pre-and post-operative care in the community will stimulate dialogue within the profession as to what is the minimal standard of care to provide patients undergoing TJA.


A network pharmacology approach to determine active compounds and action mechanisms of ge-gen-qin-lian decoction for treatment of type 2 diabetes.
Traditional Chinese medicine (TCM) herbal formulae can be valuable therapeutic strategies and drug discovery resources. However, the active ingredients and action mechanisms of most TCM formulae remain unclear. Therefore, the identification of potent ingredients and their actions is a major challenge in TCM research. In this study, we used a network pharmacology approach we previously developed to help determine the potential antidiabetic ingredients from the traditional Ge-Gen-Qin-Lian decoction (GGQLD) formula. We predicted the target profiles of all available GGQLD ingredients to infer the active ingredients by clustering the target profile of ingredients with FDA-approved antidiabetic drugs. We also applied network target analysis to evaluate the links between herbal ingredients and pharmacological actions to help explain the action mechanisms of GGQLD. According to the predicted results, we confirmed that a novel antidiabetic ingredient from Puerariae Lobatae radix (Ge-Gen), 4-Hydroxymephenytoin, increased the insulin secretion in RIN-5F cells and improved insulin resistance in 3T3-L1 adipocytes. The network pharmacology strategy used here provided a powerful means for identifying bioactive ingredients and mechanisms of action for TCM herbal formulae, including Ge-Gen-Qin-Lian decoction.


Herpes simplex virus thymidine kinase and specific stages of latency in murine trigeminal ganglia.
From marker rescue, sequencing, transcript, and latency analyses of the thymidine kinase-negative herpes simplex virus mutant dlsactk and studies using the thymidine kinase inhibitor Ro 31-5140, we infer that the virus-encoded thymidine kinase is required in murine trigeminal ganglia for acute replication and lytic gene expression, for increasing the numbers of cells expressing latency-associated transcripts, and for reactivation from latent infection.


Photodissociation dynamics of tryptophan and the implication of asymmetric photolysis.
Photodissociation of amino acid tryptophan in a molecular beam at wavelengths of 212.8 and 193 nm, corresponding to excitation to the second and third absorption bands, was investigated using multimass ion imaging techniques. The respective wavelengths also represent excitation to the edge of a positive circular dichroism band and the center of a negative circular dichroism band of L-tryptophan. Only one dissociation channel was observed at both photolysis wavelengths: C(8)NH(6)CH(2)CHNH(2)COOH-->C(8)NH(6)CH(2)+CHNH(2)COOH. Dissociation rates were found to be 1.3x10(6) and 5x10(6) s(-1) at the respective wavelengths. Comparison to theoretical calculation indicates that dissociation occurs on the ground state after internal conversion. Implication of asymmetric photolysis is discussed.


Physiology of Tuberization in Solanum tuberosum L: cis-Zeatin Riboside in the Potato Plant: Its Identification and Changes in Endogenous Levels as Influenced by Temperature and Photoperiod.
Using high pressure liquid chromatography, the cucumber cotyledon bioassay, and mass spectrometry a cytokinin isolated from Solanum tuberosum L. cv. Katahdin plant tissues has been identified as cis-zeatin riboside. Zeatin riboside (ZR) levels in plants grown under inducing conditions (28 C day and 13 C night with a 10-hour photoperiod) were significantly higher than those in plants grown under noninducing conditions (30 C day and 28 C night with an 18-hour photoperiod). The highest level of ZR was noted in below-ground tissue after 4 days exposure to inducing conditions, with tuber initiation observed after 8 days. A companion study conducted to determine the effect of ZR on in vitro tuberization of noninduced rhizomes revealed that after 1 month in culture, controls exhibited 0% tuberization, while ZR treatments of 0.3 and 3.0 milligrams per liter showed 39 and 75% tuberization, respectively.


An assay for colony-stimulating factor (CSF) production by single T lymphocytes: estimation of the frequency of cells producing granulocyte-macrophage CSF and multi-lineage CSF within a T lymphocyte clone.
The frequency of cells producing hemopoietic colony-stimulating factors (CSF) in a murine T lymphocyte clone has been determined by using a simple microassay that does not require clonal expansion or the addition of accessory cells. When stimulated with concanavalin A (Con A), the clone LB3 produced both granulocyte-macrophage CSF (GM-CSF) and multi-lineage CSF (Multi-CSF), which could be detected by using the cell line FDC-P1, whose proliferation is dependent on the presence of either of these factors. Limiting dilution analysis of Con A-stimulated LB3 cells indicated a requirement for cell-cell contact for optimal production of CSF, which could be bypassed by preincubation of the cells at high density with Con A for 4 hr before dilution in the assay. Limiting dilution estimates of the frequency of CSF-producing cells among Con A-pretreated LB3 cells ranged from 20 to 50%. Direct measurement of CSF production by single Con A-pretreated cells isolated by micromanipulation revealed that 10 to 20% could secrete detectable CSF. However, when isolated Con A-pretreated two-cell and three-cell aggregates were assayed, 50 to 99% were positive, indicating that 30 to 80% of the cells in the aggregates secreted CSF. Assay of the supernatants from single cells and two-cell aggregates on both FDC-P1 cells and another cell line, 32D c13, which responds only to Multi-CSF, demonstrated that many cells produced GM-CSF only, and others varied in the relative quantities of GM-CSF and Multi-CSF produced.