Diagnostic Efficacy, Sensitivity, and Specificity of Xpert MTB/RIF Assay for Spinal Tuberculosis and Rifampicin Resistance.
A cross-sectional observational study. This study aims to determine the diagnostic accuracy, sensitivity, and specificity of the Xpert MTB/RIF assay (Mycobacterium Tuberculosis/Rifampicin resistance) for the detection of spinal Tuberculosis (TB) and rifampicin (RIF) resistance. The Spinal TB is often a paucibacillary extra pulmonary tuberculosis which gives a major challenge in early diagnosis and initializing the correct anti-tubercular treatment (ATT). Due to its rapidity and sensitivity, the dependence and reliability on the Xpert MTB/RIF assay has increased in the last few years. The studies describing accuracy of the Xpert MTB/RIF assay in spinal TB are scanty. This institutional review board-approved study included 360 diagnosed spinal TB patients. To determine the accuracy of the Xpert MTB/RIF assay, it was compared with other diagnostic tests like histopathology, acid fast bacilli (AFB) smear, culture, and drug sensitivity testing (DST). The Xpert MTB/RIF assay showed 86.3% sensitivity and 85.3% specificity when compared with culture for the diagnosis of Spinal TB and showed 75.86% sensitivity, 96.12% specificity for RIF resistance when compared to DST. Four cases were false positive and 11 cases were false negative for RIF resistance on the Xpert MTB/RIF assay. The Xpert MTB/RIF assay is an efficient technique for the rapid diagnosis of spinal TB; however, a clinician should not solely rely on it for starting ATT. As there are false results also with this test which should be read cautiously and be well correlated with culture and DST pattern to guide the start of sensitive drug regimen only. The purpose is to prevent exposure of the second line drugs to false cases found on the Xpert MTB/RIF assay and avoid emergence of new acquired drug resistance. 4.


Antagonistic interactions between an invasive alien and a native coccinellid species may promote coexistence.
Despite the capacity of invasive alien species to alter ecosystems, the mechanisms underlying their impact remain only partly understood. Invasive alien predators, for example, can significantly disrupt recipient communities by consuming prey species or acting as an intraguild predator (IGP). Behavioural interactions are key components of interspecific competition between predators, yet these are often overlooked invasion processes. Here, we show how behavioural, non-lethal IGP interactions might facilitate the establishment success of an invading alien species. We experimentally assessed changes in feeding behaviour (prey preference and consumption rate) of native UK coccinellid species (Adalia bipunctata and Coccinella septempunctata), whose populations are, respectively, declining and stable, when exposed to the invasive intraguild predator, Harmonia axyridis. Using a population dynamics model parameterized with these experimental data, we predicted how intraguild predation, accommodating interspecific behavioural interactions, might impact the abundance of the native and invasive alien species over time. When competing for the same aphid resource, the feeding rate of A. bipunctata significantly increased compared to the feeding in isolation, while the feeding rate of H. axyridis significantly decreased. This suggests that despite significant declines in the UK, A. bipunctata is a superior competitor to the intraguild predator H. axyridis. In contrast, the behaviour of non-declining C. septempunctata was unaltered by the presence of H. axyridis. Our experimental data show the differential behavioural plasticity of competing native and invasive alien predators, but do not explain A. bipunctata declines observed in the UK. Using behavioural plasticity as a parameter in a population dynamic model for A. bipunctata and H. axyridis, coexistence is predicted between the native and invasive alien following an initial period of decline in the native species. We demonstrate how empirical and theoretical techniques can be combined to understand better the processes and consequences of alien species invasions for native biodiversity.


Physiological activity of newly differentiated olfactory receptor neurons correlated with morphological recovery from olfactory nerve section in the salamander.
1. Extracellular unitary recordings were made from the olfactory epithelium of the salamander, Ambystoma tigrinum, at numerous time points following olfactory nerve section. Unitary response properties were correlated with histological examination of the same tissues. 2. At 10 days following nerve section, unitary activity was rarely recorded in all regions of the epithelium. Histological examination indicated that virtually the entire mature olfactory receptor cell population had undergone retrograde degeneration. Transneuronal degeneration was not observed in the olfactory bulb, although the olfactory nerve and glomerular layers were substantially reduced in size. 3. At subsequent times, unitary impulse activity gradually returned, consisting of both spontaneous activity and odor-evoked discharges. Anatomical recovery of the olfactory epithelium preceded that of the olfactory bulb. A positive correlation was found between neuronal differentiation in the olfactory epithelium and the recovery of receptor cell function. 4. Patterns of spontaneous activity, odor specificities, intensity-response functions, and adaptive properties studied in newly differentiated olfactory receptor neurons were indistinguishable from those observed in control units. This indicated that these properties were intrinsic to the receptor neurons. 5. Spontaneously active and responsive units were encountered prior to olfactory nerve connection with the bulb. It is concluded that receptor neurons pass through two phases of functional maturity: the first independent of bulbar contact and the second dependent on presumed synaptic contact with bulbar neurons.


A 25-year-old man with a large hepatic tumor and multiple nodular lesions.
A 25-year-old man without a prior history of liver disease presented with an 18-cm tumor of the right hepatic lobe, which was associated with multiple nodular lesions in the remaining parenchyma. The histologic and immunohistochemical features of the neoplasm were those of a poorly differentiated leiomyosarcoma with epithelioid features. The nodular lesions measured up to 4 cm in greatest dimension and had gross and microscopic features of focal nodular hyperplasia. This case suggests that multiple focal nodular hyperplasia may occasionally represent a parenchymal reaction to abnormal blood supply developing within the liver as a consequence of an enlarging malignant tumor.


Effect of long-term treatment with low doses of the LHRH antagonist Cetrorelix on pituitary receptors for LHRH and gonadal axis in male and female rats.
Our previous studies showed that treatment of female rats with large doses of Cetrorelix, an antagonist of luteinizing hormone-releasing hormone (LHRH), reduces levels of serum LH, estradiol, progesterone, and the concentration of pituitary LHRH receptors (LHRH-Rs) and their mRNA expression. Serum LH and testosterone levels and pituitary LHRH-R in male rats are also decreased by high doses of Cetrorelix. This approach can be used for therapy of sex hormone-dependent cancers. However, in conditions where an incomplete hormone deprivation is indicated, lower doses of Cetrorelix may suffice. Thus, we investigated the effect of a 30-day treatment with a low-dose depot formulation of Cetrorelix (20-24 microg per kg per day) on the pituitary-gonadal axis of male and female rats. In both sexes, lower serum LH levels were observed on day 4 after administration. In males, LH returned to control levels by day 10, whereas in females, a rebound LH elevation occurred. Testosterone levels in male rats were decreased up to day 20, but on day 30, the values were similar to controls. In females, serum estradiol was reduced on day 4; however, by day 10 it returned to normal. Progesterone levels were diminished through the entire period. Female rats showed diestrous smears during the first week of treatment and prolonged estrous periods thereafter. The weights of testes and ovaries were significantly lower, but not the weights of prostate, seminal vesicles, and uterus. Pituitary LHRH-R mRNA and LHRH-R protein levels were not significantly different from the controls. Thus, the treatment with low doses of Cetrorelix did not seriously impair gonadal functions. The results suggest that Cetrorelix in low doses induces only a partial pituitary-gonadal inhibition and might be indicated for treatment of endometriosis, leiomyomas, and benign prostatic hyperplasia.


Relevance of the Agency for Healthcare Research and Quality Patient Safety Indicators for children's hospitals.
Patient safety indicators (PSIs) were developed by the Agency for Healthcare Research and Quality. Our objectives were (1) to apply these algorithms to the National Association of Children's Hospitals and Related Institutions (NACHRI) Aggregate Case Mix Comparative Database for 1999-2002, (2) to establish mean rates for each of the PSI events in children's hospitals, (3) to investigate the inadequacies of PSIs in relation to pediatric diagnoses, and (4) to express the data in such a way that children's hospitals could use the PSIs determined to be appropriate for pediatric use for comparison with their own data. In addition, we wanted to use the data to set priorities for ongoing clinical investigations and to propose interventions if the indicators demonstrated preventable errors. The Agency for Healthcare Research and Quality PSI algorithms (version 2.1, revision 1) were applied to children's hospital administrative data (1.92 million discharges) from the NACHRI Aggregate Case Mix Comparative Database for 1999-2002. Rates were measured for the following events: complications of anesthesia, death in low-mortality diagnosis-related groups (DRGs), decubitus ulcer, failure to rescue (ie, death resulting from a complication, rather than the primary diagnosis), foreign body left in during a procedure, iatrogenic pneumothorax, infection attributable to medical care (ie, infections related to surgery or device placement), postoperative hemorrhage or hematoma, postoperative pulmonary embolism or venous thrombosis, postoperative wound dehiscence, and accidental puncture/laceration. Across the 4 years of data, the mean risk-adjusted rates of PSI events ranged from 0.01% (0.1 event per 1000 discharges) for a foreign body left in during a procedure to 14.0% (140 events per 1000 discharges) for failure to rescue. Review of International Classification of Diseases, Ninth Revision, Clinical Modification codes associated with each PSI category showed that the failure to rescue and death in low-mortality DRG indicators involved very complex cases and did not predict preventable events in the majority of cases. The PSI for infection attributable to medical care appeared to be accurate the majority of the time. Incident risk-adjusted rates of infections attributable to medical care averaged 0.35% (3.5 events per 1000 discharges) and varied up to fivefold from the lowest rate to the highest rate. The highest rates were up to 1.8 times the average. PSIs derived from administrative data are indicators of patient safety concerns and can be relevant as screening tools for children's hospitals; however, cases identified by these indicators do not always represent preventable events. Some, such as a foreign body left in during a procedure, iatrogenic pneumothorax, infection attributable to medical care, decubitus ulcer, and venous thrombosis, seem to be appropriate for pediatric care and may be directly amenable to system changes. Evidence-based practices regarding those particular indicators that have been reported in the adult literature need to be investigated in the pediatric population. In their present form, 2 of the indicators, namely, failure to rescue and death in low-mortality DRGs, are inaccurate for the pediatric population, do not represent preventable errors in the majority of pediatric cases, and should not be used to estimate quality of care or preventable deaths in children's hospitals. The PSIs can assist institutions in prioritizing chart review-based investigations; if clusters of validated events emerge in reviews, then improvement activities can be initiated. Large aggregate databases, such as the NACHRI Case Mix Database, can help establish mean rates of potential pediatric events, giving children's hospitals a context within which to examine their own data.


Circadian clock core component Bmal1 dictates cell cycle rhythm of proliferating hepatocytes during liver regeneration.
After partial hepatectomy (PH), the majority of remnant hepatocytes synchronously enter and rhythmically progress through the cell cycle for three major rounds to regain lost liver mass. Whether and how the circadian clock core component Bmal1 modulates this process remains elusive. We performed PH on Bmal1+/+ and hepatocyte-specific Bmal1 knockout (Bmal1hep-/-) mice and compared the initiation and progression of the hepatocyte cell cycle. After PH, Bmal1+/+ hepatocytes exhibited three major waves of nuclear DNA synthesis. In contrast, in Bmal1hep-/- hepatocytes, the first wave of nuclear DNA synthesis was delayed by 12 h, and the third such wave was lost. Following PH, Bmal1+/+ hepatocytes underwent three major waves of mitosis, whereas Bmal1hep-/- hepatocytes fully abolished mitotic oscillation. These Bmal1-dependent disruptions in the rhythmicity of hepatocyte cell cycle after PH were accompanied by suppressed expression peaks of a group of cell cycle components and regulators and dysregulated activation patterns of mitogenic signaling molecules c-Met and epidermal growth factor receptor. Moreover, Bmal1+/+ hepatocytes rhythmically accumulated fat as they expanded following PH, whereas this phenomenon was largely inhibited in Bmal1hep-/- hepatocytes. In addition, during late stages of liver regrowth, Bmal1 absence in hepatocytes caused the activation of redox sensor Nrf2, suggesting an oxidative stress state in regenerated liver tissue. Collectively, we demonstrated that during liver regeneration, Bmal1 partially modulates the oscillation of S-phase progression, fully controls the rhythmicity of M-phase advancement, and largely governs fluctuations in fat metabolism in replicating hepatocytes, as well as eventually determines the redox state of regenerated livers.NEW & NOTEWORTHY We demonstrated that Bmal1 centrally controls the synchronicity and rhythmicity of the cell cycle and lipid accumulation in replicating hepatocytes during liver regeneration. Bmal1 plays these roles, at least in part, by ensuring formation of the expression peaks of cell cycle components and regulators, as well as the timing and levels of activation of mitogenic signaling molecules.


Chronic alcohol-induced liver injury and oxidant stress are decreased in cytochrome P4502E1 knockout mice and restored in humanized cytochrome P4502E1 knock-in mice.
A major pathway for chronic ethanol-induced liver injury is ethanol-induced oxidant stress. Several pathways contribute to mechanisms by which ethanol induces oxidant stress. Although some studies support a role for cytochrome P450 2E1 (CYP2E1), others do not. Most previous studies were conducted in the intragastric infusion model of ethanol administration. There is a need to develop oral models of significant liver injury and to evaluate the possible role of CYP2E1 in ethanol actions in such models. We evaluated chronic ethanol-induced liver injury, steatosis, and oxidant stress in wild-type (WT) mice, CYP2E1 knock out (KO) mice, and humanized CYP2E1 knock-in (KI) mice, in which the human 2E1 was added back to mice deficient in the mouse 2E1. WT mice and the CYP2E1 KO and KI mice (both provided by Dr. F. Gonzalez, National Cancer Institute) were fed a high-fat Lieber-DeCarli ethanol liquid diet for 3weeks; pair-fed controls received dextrose. Ethanol produced fatty liver and oxidant stress in WT mice but liver injury (transaminases, histopathology) was minimal. Ethanol-induced steatosis and oxidant stress were blunted in the KO mice (no liver injury) but restored in the KI mice. Significant liver injury was produced in the ethanol-fed KI mice, with elevated transaminases, necrosis, and increased levels of collagen type 1 and smooth muscle actin. This liver injury in the KI mice was associated with elevated oxidant stress and elevated levels of the human CYP2E1 compared to levels of the mouse 2E1 in WT mice. Activation of JNK and decreased levels of Bcl-2 and Bcl-XL were observed in the ethanol-fed KI mice compared to the other groups. Fatty liver in the WT and the KI mice was associated with lower levels of PPARα and acyl-CoA oxidase. No such changes were found in the ethanol-fed KO mice. These results show that CYP2E1 plays a major role in ethanol-induced fatty liver and oxidant stress. It is the absence of CYP2E1 in the KO mice that is responsible for the blunting of steatosis and oxidant stress because restoring the CYP2E1 restores the fatty liver and oxidant stress. Moreover, it is the human CYP2E1 that restores these effects of ethanol, which suggests that results for fatty liver and oxidant stress from rodent models of ethanol intake and mouse CYP2E1 can be extrapolated to human models of ethanol intake and to human CYP2E1.


Proto-oncogenes, growth factor genes, receptor genes, differentiation genes and structural rearrangements in human cancer.
Information on chromosomal structural rearrangements in leukemia and solid tumors was retracted from a computerized databank and the literature. A binomial test procedure was used to determine the statistical significance of the observed numbers of breaks. Some 131 human cancer-specific chromosomal structural rearrangements were also taken from the literature. The results showed that breakage and deletion occurred preferentially in bands known to contain proto-oncogenes, growth factor genes, receptor genes and differentiation genes. Therefore, chromosomal structural rearrangements could be used to find and map new genes involved in the genesis and/or progression of neoplasia.


Home artificial nutrition in advanced cancer.
Attitudes to home artificial nutrition (HAN) in cancer vary greatly from country to country. A 6-year prospective survey of the practice of HAN in advanced cancer patients applied by a hospital-at-home programme in an Italian health district was performed to estimate the utilization rate, to evaluate efficacy in preventing death from cachexia, maintaining patients at home without burdens and distress and improving patients' performance status, and to obtain information about costs. Patients were eligible for HAN when all the following were present: hypophagia; life expectancy 6 weeks or more, suitable patient and family circumstances; and verbal informed consent. From July 1990 to June 1996, 587 patients were evaluated; 164 were selected for HAN (135 enteral and 29 parenteral) and were followed until 31 December 1996. The incidence of HAN per million inhabitants was 18.4 in the first year of activity and 33.2-36.9 in subsequent years, being 4-10 times greater than rates reported by the Italian HAN registers. On 31 December 1996, 158 patients had died because of the disease and 6 were on treatment. Mean survival was 17.2 weeks for those on enteral nutrition and 12.2 weeks for those on parenteral nutrition. Prediction of survival was 72% accurate. 95 patients had undergone 155 readmissions to hospital, where they spent 15-23% of their survival time. Burdens due to HAN were well accepted by 124 patients, an annoyance or scarcely tolerable in the remainder. The frequency of major complications of parenteral nutrition was 0.67 per year for catheter sepsis and 0.16 per year for deep vein thrombosis. Karnofsky performance score increased in only 13 patients and body weight increased in 43. The fixed direct costs per patient-day (in European Currency Units) were 14.2 for the nutrition team, 18.2 for enteral nutrition and 61 for parenteral nutrition. The results indicate that definite entry criteria and local surveys are required for the correct use of HAN in advanced cancer patients, that HAN can be applied without causing additional burdens and distress, and that its costs are not higher than hospital costs.


In vitro inhibition studies of two isozymes of human liver cytochrome P-450. Mephenytoin p-hydroxylase and sparteine monooxygenase.
Human liver preparations were used to screen various drugs for their capability of binding to mephenytoin p-hydroxylase and sparteine monooxygenase, two cytochrome P-450-catalyzed activities that are independently heritable. For this screening, any indication of competitive inhibition by the drug was interpreted as an indication of binding. Among 64 drugs and alkaloids tested, 24 compounds caused inhibition of mephenytoin p-hydroxylation but the inhibition was weak in most cases; by contrast, 40 of the 64 compounds inhibited sparteine oxidation, the inhibition being potent in many cases. The only fairly strong inhibitors of mephenytoin p-hydroxylation were the alkaloid papaverine and the monoamine oxidase inhibitors tranylcypromine and nialamide. The results of these inhibition studies confirm the independence of the two monogenic defects observed in different populations. Metabolism is possibly altered in poor metabolizers of mephenytoin with fewer drugs than in poor metabolizers of sparteine.


High pressure represses expression of the malB operon in Escherichia coli.
The formation of plaques by lambda phage in Escherichia coli was prevented by elevated hydrostatic pressure; phage plaques were not detected at 30 MPa. Furthermore, using promoter fragments derived from the malB operon, we showed that gene expression initiated from both promoters (malK-lamB and malEFG) was repressed by elevated hydrostatic pressure. Our findings suggest that high pressure affects gene expression directed by the malB regulatory interval, and this may cause a decrease in the quantities of lambda receptor protein, LamB.


Novel targeted strategies to overcome resistance in small-cell lung cancer: focus on PARP inhibitors and rovalpituzumab tesirine.
Introduction: Small-cell lung cancer (SCLC) is a highly aggressive neuroendocrine tumour, and its outcome is strongly conditioned by the rapid onset of resistance to conventional chemotherapeutics. First-line treatment with a combination of platinum agents and topoisomerase inhibitors has been the standard of care for over 30 years, with disappointing clinical outcome caused by early-acquired chemoresistance. In this disheartening scenario, novel treatment strategies are being implemented in order to either revert or bypass resistance mechanisms. Areas covered: The general mechanism of action of the standard frontline treatment regimens for SCLC, as well as the known resistance mechanisms to these drugs, is reviewed. Moreover, we focus on the current preclinical and clinical evidence on the potential role of PARP inhibitors and rovalpituzumab tesirine (Rova-T) to tackle chemoresistance in SCLC. Expert opinion: Preliminary evidence supports PARP inhibitors and Rova-T as two promising approaches to either revert or bypass chemoresistance in SCLC, respectively. The identification of potential predictive biomarkers of response to these innovative treatments (SLFN11 and DLL3) has shortened the gap between SCLC and personalized targeted therapy. Further large-scale clinical studies are urgently needed for a better designation of PARP inhibitors and Rova-T in the therapeutic algorithm of SCLC patients.


Predicting the Risk of Developing Type 1 Diabetes Using a One-Week Continuous Glucose Monitoring Home Test With Classification Enhanced by Machine-Learning: An Exploratory Study.
Detection of two or more autoantibodies (Ab) in the blood might describe those individuals at increased risk of developing type 1 diabetes (T1D) during the following years. The aim of this exploratory study is to propose a high versus low T1D risk classifier using machine-learning technology based on continuous glucose monitoring (CGM) home data. Forty-two healthy relatives of people with T1D with mean ± SD age of 23.8 ± 10.5 years, HbA1c (glycated hemoglobin) of 5.3% ± 0.3%, and BMI (body mass index) of 23.2 ± 5.2 kg/m2 with zero (low risk; N = 21), and ≥2 (high risk; N = 21) Ab, were enrolled in an NIH (National Institutes of Health)-funded TrialNet ancillary study. Participants wore a CGM for a week and consumed three standardized liquid mixed meals (SLMM) instead of three breakfasts. Glycemic features were extracted from two-hour post-SLMM CGM traces, compared across groups, and used in four supervised machine-learning Ab risk status classifiers. Recursive Feature Elimination (RFE) algorithm was used for feature selection; classifiers were evaluated through 10-fold cross-validation, using the receiver operating characteristic area under the curve (AUC-ROC) to select the best classification model. The percent time of glucose >180 mg/dL (T180), glucose range, and glucose CV (coefficient of variation) were the only significant differences between the glycemic features in the two groups with P values of .040, .035, and .028 respectively. The linear SVM (Support Vector Machine) model with RFE features achieved the best performance of classifying low-risk versus high-risk individuals with AUC-ROC = 0.88. A machine-learning technology, combining a potentially self-administered one-week CGM home test, has the potential to reliably assess the T1D risk.


Effect of Casein Phosphopeptide-amorphous Calcium Phosphate and Calcium Sodium Phosphosilicate on Artificial Carious Lesions: An in vitro Study.
To compare new remineralizing agents calcium sodium phosphosilicate paste and casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) paste with that of fluoridated toothpaste in remineralization of early carious lesions using scanning electron microscopy and energy-dispersive X-ray (SEM-EDX) analysis. Sixty sound extracted premolars were collected and placed in demineralizing solution for 4 days to produce artificial carious lesions. All specimens were evaluated for any loss of mineral content using SEM-EDX analysis. Samples were randomly assigned to three groups: Group I: Fluoridated toothpaste (control), group II: CPP-ACP paste, and group III: Calcium sodium phosphosilicate paste. Specimens were then treated with above-mentioned remineralizing agents and again measured for mineral content using SEM-EDX analysis. Group III (calcium sodium phosphosilicate paste) showed highest significant difference followed in descending order by group II (CPP-ACP paste) and group I (fluoridated toothpaste). Calcium sodium phosphosilicate paste showed maximum remineralizing potential compared with CPP-ACP and fluoridated toothpastes. Chaudhary I, Tripathi AM, Yadav G, Saha S. Effect of Casein Phosphopeptide-amorphous Calcium Phosphate and Calcium Sodium Phosphosilicate on Artificial Carious Lesions: An in vitro Study. Int J Clin Pediatr Dent 2017;10(3):261-266.


Therapeutic update on seborrheic dermatitis.
Seborrheic dermatitis is a recurrent, usually mild, skin disorder with typical clinical manifestations. As it most frequently involves exposed areas, such as the face and scalp, patients seek advice from a dermatologist in order to control their disease. This article will review the available treatments for this common dermatologic problem.


Long-term culture in matrigel enhances the insulin secretion of fetal porcine islet-like cell clusters in vitro.
To study the influence of extracellular matrix (ECM) on isolated fetal porcine islet-like cell clusters (ICC), ICC were cultured either embedded in Matrigel (MG) or growth factor-reduced Matrigel (MG-GF), or in the absence of matrix for 20 days. The insulin accumulation in the culture medium was determined every fifth day. After culture, the ICC were retrieved and used for acute insulin release experiments and subsequently subjected to either determination of DNA and insulin contents or immunohistochemistry. It was found that culture in MG and MG-GF significantly increased both the insulin accumulation in the culture medium and the insulin content/DNA compared to culture in the absence of matrix. In addition, MG culture significantly increased the insulin output with time compared with the initial 5-day value. The acute insulin release was significantly higher from the ICC cultured in the presence of matrix, whereas no acute glucose-induced insulin response could be detected. During the culture period, the ICC cultured in the absence of matrix disintegrated and formed a monolayer of cells in the culture dish, whereas the ICC cultured in matrix formed rounded structures containing a substantial number of insulin-positive cells. In some cases, cyst-like structures could be seen after culture in matrix. The data suggest that beta-cell differentiation during fetal development has a dual requirement for extracellular matrix components and growth factors.


Perspectives on health examination for asylum seekers in relation to health literacy - focus group discussions with Arabic and Somali speaking participants.
Asylum seekers coming to most countries are offered a specific health examination. A previous study concluded that a considerable proportion of those taking part of it in Sweden had poor experiences of the communication in and the usefulness of this examination and had poor health literacy. The aim of this study was to explore in greater depth the experiences of the health examination for asylum seekers among Arabic- and Somali-speaking participants in Sweden. A secondary aim was to examine experiences and discuss findings using a health literacy framework. Seven focus group discussions were conducted with 28 Arabic and Somali speaking men and women that participated in a health examination for asylum seekers. Data were analyzed by latent content analysis. One overarching theme - beneficial and detrimental - was found to represent the participants' experiences of the health examination for asylum seekers. Three categories were identified that deal with those experiences. The category of "gives some good" describes the examination as something that "gives support and relief" and "cares on a personal level." The category of "causes feelings of insecurity" describes the examination as something that "lacks clarity" and that "does not give protection." The category "causes feelings of disappointment" views the examination as something that "does not fulfil the image of a health examination" and "does not focus on the individual level." The health examination for asylum seekers was experienced as beneficial and detrimental at the same time. The feelings were influenced by the experiences of information and communication before, during and after the examination and on how health literate the organizations providing the HEA are. To achieve more satisfied participants, it is crucial that all organizations providing the HEA become health literate and person-centered.


A novel approach for addressing diseases not yielding to effective vaccination? Immunization by replication-competent controlled virus.
Vaccination involves inoculation of a subject with a disabled disease-causing microbe or parts thereof. While vaccination has been highly successful, we still lack sufficiently effective vaccines for important infectious diseases. We propose that a more complete immune response than that elicited from a vaccine may be obtained from immunization with a disease-causing virus modified to subject replication-essential genes to the control of a gene switch activated by non-lethal heat in the presence of a drug-like compound. Upon inoculation, strictly localized replication of the virus would be triggered by a heat dose administered to the inoculation site. Activated virus would transiently replicate with an efficiency approaching that of the disease-causing virus and express all viral antigens. It may also vector heterologous antigens or control co-infecting microbes.


Emergency analysis of paraquat in biological fluids.
A variety of spectrophotometric, gas and liquid chromatographic and radioimmunoassay techniques have been applied to the measurement of paraquat in biological fluids. A brief review of these techniques is presented. The majority of described methods are far from suitable in the provision of an accurate and reliable quantitative result in an emergency situation and the further development of suitable 'rapid' techniques is desirable. The preparation and characterisation of internal accuracy control materials and the introduction of an external quality assessment scheme would be valuable in the improvement of laboratory investigations of paraquat poisoning.


Squarephaneic Tetraanhydride: A Conjugated Square-Shaped Cyclophane for the Synthesis of Porous Organic Materials.
Aromatic carboxylic anhydrides are ubiquitous building blocks in organic materials chemistry and have received considerable attention in the synthesis of organic semiconductors, pigments, and battery electrode materials. Here we extend the family of aromatic carboxylic anhydrides with a unique new member, a conjugated cyclophane with four anhydride groups. The cyclophane is obtained in a three-step synthesis and can be functionalised efficiently, as shown by the conversion into tetraimides and an octacarboxylate. Crystal structures reveal the high degree of porosity achievable with the new building block. Excellent electrochemical properties and reversible reduction to the tetraanions are shown for the imides; NMR and EPR measurements confirm the global aromaticity of the dianions and evidence the global Baird aromaticity of the tetraanions. Considering the short synthesis and unique properties, we expect widespread use of the new building block in the development of organic materials.


Histogenesis of granular cell tumour--an immunohistochemical and ultrastructural study.
Fifteen cases of granular cell tumour of superficial soft tissues or tongue were immunohistochemically evaluated for different types of intermediate filament proteins and for laminin, a glycoprotein of basal laminae. Four of the tumours were studied ultrastructurally. The tumour cells appeared to contain only vimentin-type of intermediate filament protein. The lobules of tumour cells were surrounded by laminin-positive material, but in contrast to schwannomas and neurofibromas, the individual tumour cells were not covered by laminin. In line with the immunohistochemical observations, by electron microscopy basal lamina-like material could not be demonstrated between individual cells, but only surrounding groups of cells. Lysozyme, a histiocytic marker, was absent in the tumour cells. Our results do not confirm any particular cell type for the histogenetic origin of granular cell tumour, but suggest that it may rather be derived from uncommitted possibly nerve-related mesenchymal cells.


In situ vaccination combined with androgen ablation and regulatory T-cell depletion reduces castration-resistant tumor burden in prostate-specific pten knockout mice.
There is no effective treatment for prostate cancer arising after androgen ablation. Previous studies have analyzed the short-term effects of androgen ablation on the immune system and suggest an abatement of immune suppression by hormone removal. Because castration-resistant disease can arise years after treatment, it is crucial to determine the duration of immune potentiation by castration. Because immunotherapeutic efficacy is determined by the balance of immune cell subsets and their location within the tumor, we assessed the acute and chronic effect of androgen ablation on the localization of T-cell subsets within castration-resistant murine prostate cancer. We observed a transient increase in CD4+ and CD8+ T-cell numbers at the residual tumor after androgen ablation. More than 2 months later, regulatory T cells (Treg) were increasingly found within prostate epithelium, whereas CTLs, which were evenly distributed before androgen ablation, became sequestered within stroma. Anti-CD25 antibody administration along with castration enhanced CTL access to cancerous glands but did not increase effector function. Intraprostatic injection of LIGHT-expressing tumor cells increased the proportion of CD8+ T cells with functional capacity within the cancerous gland. In addition, Treg depletion within the tumor was enhanced. Together, these manipulations significantly reduced castration-resistant tumor burden. Thus, our results indicate that immune modulations, which prevent Treg accumulation and augment effector cell infiltration of prostatic epithelium, may be effective in reducing tumor burden or preventing tumor recurrence after androgen ablation therapy.


Health services utilization under Qassam rocket attacks.
The Qassam rocket attacks on southern Israel during the years 2000-2007 created a unique situation of life undera continuous threat. The effect of this unique situation on health services utilization has not been previously examined. To evaluate health utilization patterns in two primary care clinics in southern Israel: one under continuous attacks of Qassam rockets as compared with a similar clinic not under a rocket threat. We conducted a retrospective cross-sectional study in two primary care clinics in southern Israel, with 11,630 persons listed in the two clinics during the entire study period. The primary outcome measures were total annual number of visits per person to the clinic and for specific diagnoses, and the number of drug prescriptions issued, emergency room (ER) visits, hospitalization days, cardiac catheterizations and coronary bypass surgeries. In both clinics there was an increase over time in the mean annual number of visits per person. During the years of severe attacks there was an increase in visits with a chief complaint of depression and anxiety and an increase in the number of anxiolytic prescriptions in the study clinic compared with the control. During the same period there was a decrease in the number of ER visits in the study clinic compared with the control. The population under continuous life-threatening events showed more depression and anxiety problems. Under severe bombardment, the residents prefer not to leave home, unless necessary.


New insights into the Saccharomyces cerevisiae fermentation switch: dynamic transcriptional response to anaerobicity and glucose-excess.
The capacity of respiring cultures of Saccharomyces cerevisiae to immediately switch to fast alcoholic fermentation upon a transfer to anaerobic sugar-excess conditions is a key characteristic of Saccharomyces cerevisiae in many of its industrial applications. This transition was studied by exposing aerobic glucose-limited chemostat cultures grown at a low specific growth rate to two simultaneous perturbations: oxygen depletion and relief of glucose limitation. The shift towards fully fermentative conditions caused a massive transcriptional reprogramming, where one third of all genes within the genome were transcribed differentially. The changes in transcript levels were mostly driven by relief from glucose-limitation. After an initial strong response to the addition of glucose, the expression profile of most transcriptionally regulated genes displayed a clear switch at 30 minutes. In this respect, a striking difference was observed between the transcript profiles of genes encoding ribosomal proteins and those encoding ribosomal biogenesis components. Not all regulated genes responded with this binary profile. A group of 87 genes showed a delayed and steady increase in expression that specifically responded to anaerobiosis. Our study demonstrated that, despite the complexity of this multiple-input perturbation, the transcriptional responses could be categorized and biologically interpreted. By comparing this study with public datasets representing dynamic and steady conditions, 14 up-regulated and 11 down-regulated genes were determined to be anaerobic specific. Therefore, these can be seen as true "signature" transcripts for anaerobicity under dynamic as well as under steady state conditions.


Genomic Sequencing of Bacillus cereus Sensu Lato Strains Isolated from Meat and Poultry Products in South Africa Enables Inter- and Intranational Surveillance and Source Tracking.
Members of the Bacillus cereus sensu lato species complex, also known as the B. cereus group, vary in their ability to cause illness but are frequently isolated from foods, including meat products; however, food safety surveillance efforts that use whole-genome sequencing (WGS) often neglect these potential pathogens. Here, we evaluate the surveillance and source tracking potential of WGS as applied to B. cereus sensu lato by (i) using WGS to characterize B. cereus sensu lato strains isolated during routine surveillance of meat products across South Africa (n = 25) and (ii) comparing the genomes sequenced here to all publicly available, high-quality B. cereus sensu lato genomes (n = 2,887 total genomes). Strains sequenced here were collected from meat products obtained from (i) retail outlets, processing plants, and butcheries across six South African provinces (n = 23) and (ii) imports held at port of entry (n = 2). The 25 strains sequenced here were partitioned into 15 lineages via in silico seven-gene multilocus sequence typing (MLST). While none of the South African B. cereus sensu lato strains sequenced here were identical to publicly available genomes, six MLST lineages contained multiple strains sequenced in this study, which were identical or nearly identical at the whole-genome scale (≤3 core single nucleotide polymorphisms). Five MLST lineages contained (nearly) identical genomes collected from two or three South African provinces; one MLST lineage contained nearly identical genomes from two countries (South Africa and the Netherlands), indicating that B. cereus sensu lato can spread intra- and internationally via foodstuffs. IMPORTANCE Nationwide foodborne pathogen surveillance programs that use high-resolution genomic methods have been shown to provide vast public health and economic benefits. However, Bacillus cereus sensu lato is often overlooked during large-scale routine WGS efforts. Thus, to our knowledge, no studies to date have evaluated the potential utility of WGS for B. cereus sensu lato surveillance and source tracking in foodstuffs. In this preliminary proof-of-concept study, we applied WGS to B. cereus sensu lato strains collected via South Africa's national surveillance program of domestic and imported meat products, and we provide strong evidence that B. cereus sensu lato can be disseminated intra- and internationally via the agro-food supply chain. Our results showcase that WGS has the potential to be used for source tracking of B. cereus sensu lato in foods, although future WGS and metadata collection efforts are needed to ensure that B. cereus sensu lato surveillance initiatives are on par with those of other foodborne pathogens.


Next generation per- and poly-fluoroalkyl substances: Status and trends, aquatic toxicity, and risk assessment.
Widespread application of poly- and per-fluoroalkyl substances (PFAS) has resulted in some substances being ubiquitous in environmental matrices. That and their resistance to degradation have allowed them to accumulate in wildlife and humans with potential for toxic effects. While specific substances of concern have been phased-out or banned, other PFAS that are emerging as alternative substances are still produced and are being released into the environment. This review focuses on describing three emerging, replacement PFAS: perfluoroethylcyclohexane sulphonate (PFECHS), 6:2 chlorinated polyfluoroalkyl ether sulfonate (6:2 Cl-PFAES), and hexafluoropropylene oxide dimer acid (HFPO-DA). By summarizing their physicochemical properties, environmental fate and transport, and toxic potencies in comparison to other PFAS compounds, this review offers insight into the viabilities of these chemicals as replacement substances. Using the chemical scoring and ranking assessment model, the relative hazards, uncertainties, and data gaps for each chemical were quantified and related to perfluorooctane sulfonic acid (PFOS) and perfluorooctanoic acid (PFOA) based on their chemical and uncertainty scores. The substances were ranked PFOS > 6:2 Cl-PFAES > PFOA > HFPO-DA > PFECHS according to their potential toxicity and PFECHS > HFPO-DA > 6:2 Cl-PFAES > PFOS > PFOA according to their need for future research. Since future uses of PFAS remain uncertain in the face of governmental regulations and production bans, replacement PFAS will continue to emerge on the world market and in the environment, raising concerns about their general lack of information on mechanisms and toxic potencies.


Three-dimensional organization of dendrites and local axon collaterals of shell and core medium-sized spiny projection neurons of the rat nucleus accumbens.
Medium-sized spiny projection neurons (MSN) in the head of the primate caudate nucleus are thought to have preferred dendritic orientations that tend to parallel the orientations of the striosomes. Moreover, recurrent axon collaterals of MSN in the rat dorsal striatum have been categorized into two types, i.e., restricted and widespread. The nucleus accumbens (Acb) has a highly complex compartmental organization, and the spatial organization of dendritic and axonal arbors of MSN has not yet been systematically studied. In this study, using single-cell juxtacellular labeling with neurobiotin as well as anterograde neuroanatomical tracing with biotinylated dextran amine, we investigated the three-dimensional (3D) organization of dendrites and axons of MSN of the rat Acb in relation to subregional (shell-core) and compartmental (patch-matrix) boundaries. Our results show that dendritic arbors of MSN in both the Acb shell and core subregions are preferentially oriented, i.e., they are flattened in at least one of the 3D-planes. The preferred orientations are influenced by shell-core and patch-matrix boundaries, suggesting parallel and independent processing of information. Dendritic orientations of MSN of the Acb core are more heterogeneous than those of the shell and the dorsal striatum, suggesting a more complex distribution of striatal inputs within the core. Although dendrites respect the shell-core and patch-matrix boundaries, recurrent axon collaterals may cross these boundaries. Finally, different degrees of overlap between dendritic and axonal arborizations of individual MSN were identified, suggesting various possibilities of lateral inhibitory interactions within and between, functionally distinct territories of the Acb.


Quantitative Assessment of Molecular Transport through Sub-3 nm Silica Nanochannels by Scanning Electrochemical Microscopy.
Scanning electrochemical microscopy (SECM) has been proved to be a powerful technique to study molecular transport across ionic channels in biomembranes and artificial nanoporous membranes. In this work SECM was used to study the dynamics of molecular transport across the ultrathin silica nanoporous membrane consisting of sub-3 nm diameter perpendicular channels. We focused on the quantitative assessment of permselectivity and permeability of the membrane and the effect of radial electrical double layer (EDL) on them. By SECM imaging, it was phenomenologically observed that the membrane with negatively charged surface exhibited permselectivity to anionic molecule, for instance hexacyanoruthenate(II) (Ru(CN)64-). And the permselective transport of Ru(CN)64- was obviously more favored at a higher concentration of KCl. Precise membrane permeability to Ru(CN)64- was quantitatively determined by overlapping experimental SECM approach curves with the ones generated by finite element simulations. The high permeability up to 35 μm s-1 was ascribed to the straight channel structure and ultrahigh channel density of 4 × 1012 cm-2. Moreover, the permeability was varied from 35 μm s-1 to 2.5 μm s-1 when decreasing the concentration of KCl from 1.0 to 0.01 M, corroborating the electrostatic origin of membrane permselectivity. On the other hand, the simulated concentration profiles at both sides of the membrane suggested that the molecular transport across the membrane was mainly driven by the large transmembrane concentration gradient while the tip-induced transport was relatively negligible. These results help to quantitatively understand the molecular transport selectivity and dynamics across nanoporous membranes and to rationally design artificial molecular sieving membranes.


Super-enhancers maintain renin-expressing cell identity and memory to preserve multi-system homeostasis.
Renin cells are crucial for survival - they control fluid-electrolyte and blood pressure homeostasis, vascular development, regeneration, and oxygen delivery to tissues. During embryonic development, renin cells are progenitors for multiple cell types that retain the memory of the renin phenotype. When there is a threat to survival, those descendants are transformed and reenact the renin phenotype to restore homeostasis. We tested the hypothesis that the molecular memory of the renin phenotype resides in unique regions and states of these cells' chromatin. Using renin cells at various stages of stimulation, we identified regions in the genome where the chromatin is open for transcription, mapped histone modifications characteristic of active enhancers such as H3K27ac, and tracked deposition of transcriptional activators such as Med1, whose deletion results in ablation of renin expression and low blood pressure. Using the rank ordering of super-enhancers, epigenetic rewriting, and enhancer deletion analysis, we found that renin cells harbor a unique set of super-enhancers that determine their identity. The most prominent renin super-enhancer may act as a chromatin sensor of signals that convey the physiologic status of the organism, and is responsible for the transformation of renin cell descendants to the renin phenotype, a fundamental process to ensure homeostasis.


[The surgical treatment of radiation damage to the rectum].
Radiation therapy of cervical cancer causes late radiation lesions of the rectum in a majority of patients. Surgery is a method of choice of the treatment of postradiation corrosive strictures and fistulas. Modern diagnosis of complications permits adequate surgical rehabilitation.


Physiology of peripheral nerve fibres in relation to their size.
There is evidence that peripheral nerve fibres, both myelinated and unmyelinated, have different membrane properties correlated with their size. Smaller fibres have a lower conduction velocity, and longer action potential time course, than would be expected if their membrane properties were identical with those of larger size. There are several possible explanantions of this result; it is suggested that at least part of the difference is due to a systematic variation in the size of the conductances (per unit area of membrane) responsible for generating the action potential.


Adverse Plaque Characteristics Relate More Strongly With Hyperemic Fractional Flow Reserve and Instantaneous Wave-Free Ratio Than With Resting Instantaneous Wave-Free Ratio.
The current substudy of the PACIFIC (Prospective Comparison of Cardiac PET/CT, SPECT/CT Perfusion Imaging and CT Coronary Angiography With Invasive Coronary Angiography) trial explores the impact of computed tomography (CT)-derived unfavorable plaque features on both hyperemic and non-hyperemic flow indices. Next to lesion severity, plaque vulnerability as assessed using coronary CT angiography affects fractional flow reserve (FFR), which is associated with imminent acute coronary syndromes. Instantaneous wave-free ratio (iFR) has recently emerged as an alternative for FFR to interrogate coronary lesions for ischemia. It is, however, unknown whether vasodilator-free assessment with iFR is associated with plaque stability similarly as FFR. Of 120 patients (62% men, age 58.3 ± 8.6 years) with suspected coronary artery disease, 257 vessels were prospectively evaluated. Each patient underwent 256-slice coronary CT angiography to assess stenosis severity and plaque features (positive remodeling [PR], low attenuation plaque [LAP], spotty calcification [SC], and napkin ring sign [NRS]), as well as intracoronary pressure measurements (FFR, iFR, Pd/Pa, and pressure ratio during adenosine within the wave-free period [iFRa]). CT-derived plaque characteristics were related to these invasive pressure measurements. Atherosclerotic plaques were present in 170 (66%) coronary arteries. On a per-vessel basis, luminal stenosis severity was significantly associated with impaired FFR, iFR, Pd/Pa, and iFRa. Multivariable analysis revealed that FFR and iFR were independently related to ≥70% stenosis (-0.10, p < 0.001 and -0.09, p = 0.003, respectively) and plaque volume (-0.02, p = 0.020 and -0.02, p = 0.030, respectively). Additionally, PR and SC were also independent predictors of an impaired FFR (-0.10, p < 0.001 and -0.07, p = 0.021, respectively), but adverse plaque characteristics were not independently related to the vasodilator-free iFR. CT-derived vulnerable plaque characteristics are independently associated with hyperemic flow indices as assessed with FFR and iFRa, but not with non-hyperemic indices such as iFR and Pd/Pa. These findings suggest that the effects of hyperemia on pressure-derived indices might depend not only on hemodynamic stenosis severity but also on plaque characteristics.


Polyamines in intestinal growth.
The total cellular mass of the small intestine is well controlled and can adapt, with hypo- or hyperplasia, to a wide variety of stimuli. Luminal nutrients, hormonal factors and pancreatic and biliary secretions have all been implicated in the regulation of intestinal mucosal growth. The polyamines (putrescine, spermidine and spermine) and the key enzyme controlling their synthesis (ornithine decarboxylase. ODC) are critical for many cell growth processes and appear to play important roles in intestinal growth. During intestinal adaptation in response to jejunectomy, lactation. pancreatic-biliary diversion, starvation-refeeding and feeding with kidney bean lectin, intestinal contents of ODC and polyamines are increased, paralleling increases in mucosal proliferative indices and DNA synthesis. With administration of the specific inhibitor of ODC (difluoromethylornithine, DFMO) the increase in ODC and polyamines is inhibited and intestinal growth is suppressed. In addition, the oral administration of exogenous polyamines results in precocious maturation of the neonatal rat intestine. These results suggest that the polyamines are important for intestinal growth.


Causes of Iatrogenic Median Nerve Injury after Endoscopic Carpal Tunnel Release.
Background: Endoscopic carpal tunnel release (ECTR) is a less invasive procedure but has a higher risk of complications. We analysed ECTR cases dividing them into three periods according to a single surgeon's experience when the ECTR was performed: the initial, midterm and late period. Cases of iatrogenically induced median nerve injuries that occurred after ECTR were then noted and evaluated. Methods: We reviewed 195 ECTRs done with the 2-portal technique and divided the patients into three groups according to periods of when ECTR was done. The indications for ECTR surgery were limited to severe CTS cases. These groups of patients were similar in terms of age, duration of disease, electrophysiological study results and severity of the disease. The patients were evaluated for median neuropathy pre- and postoperatively using Semmes-Weinstein monofilament test (SWT), Disabilities of the arm, shoulder and hand (DASH) Score, Coin-flip test (CFT), postoperative paraesthesias and complications, such as pillar pain, and so on. Electrophysiological evaluation was performed only preoperatively. Results: Postoperative median nerve recovery was overall good. Normal recovery was noted in 181 cases (93%). SWT, DASH and CFT were all significantly improved upon follow-up in all three groups. In terms of iatrogenic neuropathy, median nerve palsy worsened (including those transiently worsened) after ECTR in 11 cases (5.6%), even in the later period. The sensory disturbance was equally worsening from the radial to the ulnar side. Conclusions: The fact that there were neurologically worsened cases even in the later period, when the operator is higher skilled in the technique, suggests that the surgical technique itself may be the one posing higher risk than the level of surgical skill. The most likely causes of aggravated nerve palsy were a direct injury by cannula insertion at the proximal portal, or additional median nerve compression during cannula insertion into the carpal tunnel. Level of Evidence: Level IV (Therapeutic).


NMP4 regulates the innate immune response to influenza A virus infection.
Severe influenza A virus infection typically triggers excessive and detrimental lung inflammation with massive cell infiltration and hyper-production of cytokines and chemokines. We identified a novel function for nuclear matrix protein 4 (NMP4), a zinc-finger-containing transcription factor playing roles in bone formation and spermatogenesis, in regulating antiviral immune response and immunopathology. Nmp4-deficient mice are protected from H1N1 influenza infection, losing only 5% body weight compared to a 20% weight loss in wild type mice. While having no effects on viral clearance or CD8/CD4 T cell or humoral responses, deficiency of Nmp4 in either lung structural cells or hematopoietic cells significantly reduces the recruitment of monocytes and neutrophils to the lungs. Consistent with fewer innate cells in the airways, influenza-infected Nmp4-deficient mice have significantly decreased expression of chemokine genes Ccl2, Ccl7 and Cxcl1 as well as pro-inflammatory cytokine genes Il1b and Il6. Furthermore, NMP4 binds to the promoters and/or conserved non-coding sequences of the chemokine genes and regulates their expression in mouse lung epithelial cells and macrophages. Our data suggest that NMP4 functions to promote monocyte- and neutrophil-attracting chemokine expression upon influenza A infection, resulting in exaggerated innate inflammation and lung tissue damage.


Military service absences and family members' mental health: A timeline followback assessment.
Although military service, and particularly absence due to deployment, has been linked to risk for depression and anxiety among some spouses and children of active duty service members, there is limited research to explain the heterogeneity in family members' reactions to military service stressors. The current investigation introduces the Timeline Followback Military Family Interview (TFMFI) as a clinically useful strategy to collect detailed time-linked information about the service member's absences. Two dimensions of parent absence--the extent to which absences coincide with important family events and cumulative time absent--were tested as potential risks to family members' mental health. Data from 70 mother-adolescent pairs revealed that the number of important family events missed by the service member was linked to elevated youth symptoms of depression, even when accounting for the number of deployments and cumulative duration of the service member's absence. However, youth who reported more frequent contact with the service member during absences were buffered from the effects of extensive absence. Mothers' symptoms were associated with the cumulative duration of the service members' time away, but not with family events missed by the service member. These results identify circumstances that increase the risk for mental health symptoms associated with military family life. The TFMFI provides an interview-based strategy for clinicians wishing to understand military family members' lived experience during periods of service-member absence.


Acquired retinal folds in the cat.
Retinal folds were found in 5 cats. The apparent cause of the folding was varied: in 1 cat the folds appeared after a localized retinal detachment; in 2 cats the condition accompanied other intraocular abnormalities associated with feline infectious peritonitis; 1 cat had active keratitis, and the retinal changes were thought to have been injury related; and 1 cat, bilaterally affected, had chronic glomerulonephritis.


Lateralized processing of emotional images: A left hemisphere memory deficit.
The right hemisphere plays a critical role in visuospatial attention and emotional perception, particularly for negative emotions. Therefore, preferential processing of emotional stimuli by the right, compared to the left, hemisphere could enhance our memory for emotional stimuli. We examined whether recognition memory for negative versus neutral IAPS images (Experiment 1) and negative versus positive IAPS and NAPS images (Experiment 2) differed depending on initial right or left hemisphere processing-manipulated by presenting images in the left (i.e., right hemisphere) or the right (i.e., left hemisphere) visual field. We tested recognition memory for valence-matched image pairs encoded for 500 ms. We manipulated image valence and visual field of presentation within-participants. In Experiment 1, valence and visual field interacted to influence recognition memory: people recognized negative images encoded from the left visual field (right hemisphere) more accurately than negative images encoded from the right visual field (left hemisphere). There were no visual field differences for neutral images. In Experiment 2, recognition memory for positive and negative images was equally enhanced when these images appeared in the left compared to the right visual field. Our findings suggest preferential right hemisphere processing of visuospatial and emotional information enhances recognition memory for emotional images. We interpret these findings as a left hemisphere memory deficit for emotional images, because right hemisphere processing led to similar recognition memory of emotional as neutral images, whereas left hemisphere processing led to worse recognition memory of emotional than neutral images. These findings aid our understanding of how lateralized functions contribute to emotional processing. (PsycINFO Database Record (c) 2020 APA, all rights reserved).


Transfer of murine leukaemia and murine sarcoma virus genetic information by transfection with isolated metaphase chromosomes.
Chromosome-mediated transfer of murine leukaemia (MuLV) and murine sarcoma (MuSV) virus genetic information to uninfected recipient cells was investigated. Metaphase chromosomes from AKR MuLV-infected SC-1 mouse cells were incubated with NIH/3T3 cells. After several passages (1 to 3 weeks), infectious virions exhibiting reverse transcriptase activity and the characteristic host range of ecotropic, N-tropic AKR virus appeared in the supernatant fluids of the treated cells. Restriction endonuclease analysis of genomic DNA from transfected cells indicated that AKR proviral DNA was associated with the high molecular weight DNA of the host. These results demonstrate that the AKR MuLV genome can be stably transferred to uninfected recipient cells via isolated metaphase chromosomes. Although AKR virions are not able to infect heterologous cells, chromosome-mediated transfection resulted in the establishment of productive AKR MuLV infection in mink cells. Thus, the use of chromosomes to transfer virus genes can circumvent the natural host restriction barrier. In other experiments, it was shown that normal NIH/3T3 cells were transformed after exposure to metaphase chromosomes isolated from an MuSV-infected, non-producer line. Foci were detected 14 to 21 days after chromosome treatment and were shown to contain true viral transformants since transforming virus was produced after superinfection with MuLV.


Survival and failure pattern of patients with endometrial cancer after extensive surgery including systematic pelvic and para-aortic lymphadenectomy followed by adjuvant chemotherapy.
We investigated the survival and the failure pattern of 288 patients with endometrial cancer treated with extensive surgery including systematic pelvic and para-aortic lymphadenectomy followed by cisplatin-based chemotherapy from 1982 to 2002. We correlated the failure pattern with various clinicopathologic factors to find the predictors of recurrence sites. The 5-year overall survival rates were 97.5% for stage I, 87.5% for stage II, 85.2% for stage III, and 12.5% for stage IV. Notably, the 5-year survival rate was 76.5% for patients with stage IIIC disease. Among patients with a low risk (n = 92) for recurrence who received no adjuvant chemotherapy, 2 (2.2%) showed recurrent disease. Among those with intermediate (n = 98) and high (n = 98) risks for recurrence who received adjuvant chemotherapy, 9 (9.2%) and 20 (20.4%) showed recurrent disease, respectively. The recurrence sites were described as follows: distant (n = 12), vaginal (n = 8), peritoneal (n = 7), pelvic (n = 2), and lymphatic (n = 2). Lymphatic failure was found beyond the area of lymphadenectomy. Architectural and nuclear grades; myometrial, lymph-vascular space, and cervical invasions; and lymph node metastasis were predictors of distant failure. Cervical invasion and lymph node metastasis were predictors of vaginal failure. For patients with stage I/II cancer, the architectural and nuclear grades were related to distant failure. Seven (63.6%) of 11 patients with a low or intermediate risk survived after relapse, whereas only 1 (4.8%) of 21 patients with a high risk survived after a recurrence. We conclude that we need to further test the efficacy of systemic adjuvant therapy using new chemotherapeutic regimens to prevent distant failure and to improve the survival of patients with endometrial cancer.


Identification, selection, and expansion of non-gene modified alloantigen-reactive Tregs for clinical therapeutic use.
Transplantation is limited by the need for life-long pharmacological immunosuppression, which carries significant morbidity and mortality. Regulatory T cell (Treg) therapy holds significant promise as a strategy to facilitate immunosuppression minimization. Polyclonal Treg therapy has been assessed in a number of Phase I/II clinical trials in both solid organ and hematopoietic transplantation. Attention is now shifting towards the production of alloantigen-reactive Tregs (arTregs) through co-culture with donor antigen. These allospecific cells harbour potent suppressive function and yet their specificity implies a theoretical reduction in off-target effects. This review will cover the progress in the development of arTregs including their potential application for clinical use in transplantation, the knowledge gained so far from clinical trials of Tregs in transplant patients, and future directions for Treg therapy.


[Informatics competencies essential to decision making in nursing management].
To identify informatics abilities essential to decision making in nursing management. Survey study with specialist nurses in health informatics and management. An electronic questionnaire was built based on the competencies Information Literacy (five categories; 40 abilities) and Information Management (nine categories; 69 abilities) of the TIGER - Technology Informatics Guiding Education Reform - initiative, with the guiding question: Which informatics abilities are essential to decision making in management? Answers were sorted in a Likert scale, ranging from 1 to 5. Rasch analysis was conducted with the software WINSTEPS(®). Results were presented in logits, with cutoff value zero. Thirty-two specialists participated, coming from all regions of Brazil. In the information literacy competency, 18 abilities were considered essential and in Information Management, 38; these were sorted according to their degree of essentiality. It is believed that the incorporation of these abilities in teaching can support the education of nurse managers and contribute to evidence-based practice, incorporation of information and communication technologies in health and information management.


Peg-hole disassembly using active compliance.
When considered in two-dimensional space, a cylindrical peg being withdrawn from a clearance-fit hole can exhibit one of four contact states: no contact, one-point contact, two-point contact and line contact. Jamming and wedging can occur during the two-point contact. Effective control of the two-point contact region can significantly reduce resistance in peg-hole disassembly. In this paper, we explore generic peg-hole disassembly processes with compliance and identify the effects of key parameters including the degree of compliance, the location of the compliance centre and initial position errors. A quasi-static analysis of peg-hole disassembly has been performed to obtain the boundary conditions of the two-point contact region. The effects of key variables on the two-point contact region have been simulated. Finally, peg-hole disassemblies with different locations of compliance centre achieved using active compliance have been experimentally investigated. The proposed theoretical model can be implemented to predict the range and position of the two-point contact region from the perspective of peg-hole disassembly.


Do motives matter in male circumcision? 'Conscientious objection' against the circumcision of a Muslim child with a blood disorder.
Whilst there have been serious attempts to locate the practice of male circumcision for religious motives in the context of the (respective) religion's narrative and community, the debate, when referring to a clinical context, is often more nuanced. This article will contribute further to the debate by contextualising the Islamic practice of male circumcision within the clinical setting typical of a contemporary hospital. It specifically develops an additional complication; namely, the child has a pre-existing blood disorder. As an approach to contributing to the circumcision debate further, the ethics of a conscientious objection for secular motives towards a religiously-motivated clinical intervention will be explored. Overall, the discussion will provide relevance for such debates within the value-systems of a multi-cultural society. This article replicates several approaches to deconstructing a request for conscientious refusal of non-therapeutic circumcision by a Clinical Ethics Committee (CEC), bringing to light certain contradictions that occur in normatively categorizing motives for performing the circumcision.


Towards building equitable health systems in Sub-Saharan Africa: lessons from case studies on operational research.
Published practical examples of how to bridge gaps between research, policy and practice in health systems research in Sub Saharan Africa are scarce. The aim of our study was to use a case study approach to analyse how and why different operational health research projects in Africa have contributed to health systems strengthening and promoted equity in health service provision. Using case studies we have collated and analysed practical examples of operational research projects on health in Sub-Saharan Africa which demonstrate how the links between research, policy and action can be strengthened to build effective and pro-poor health systems. To ensure rigour, we selected the case studies using pre-defined criteria, mapped their characteristics systematically using a case study development framework, and analysed the research impact process of each case study using the RAPID framework for research-policy links. This process enabled analysis of common themes, successes and weaknesses. 3 operational research projects met our case study criteria: HIV counselling and testing services in Kenya; provision of TB services in grocery stores in Malawi; and community diagnostics for anaemia, TB and malaria in Nigeria. Political context and external influences: in each case study context there was a need for new knowledge and approaches to meet policy requirements for equitable service delivery. Collaboration between researchers and key policy players began at the inception of operational research cycles. Links: critical in these operational research projects was the development of partnerships for capacity building to support new services or new players in service delivery. evidence was used to promote policy dialogue around equity in different ways throughout the research cycle, such as in determining the topic area and in development of indicators. Building equitable health systems means considering equity at different stages of the research cycle. Partnerships for capacity building promotes demand, delivery and uptake of research. Links with those who use and benefit from research, such as communities, service providers and policy makers, contribute to the timeliness and relevance of the research agenda and a receptive research-policy-practice interface. Our study highlights the need to advocate for a global research culture that values and funds these multiple levels of engagement.


Cerebral sparganosis mansoni--case report.
An 80-year-old male with paresis of the right upper extremity underwent surgery for a suspected metastatic brain tumor. A live Sparganum mansoni worm was removed from the parietal lobe through a left parietal craniotomy. Retrospective evaluation of magnetic resonance (MR) images revealed ipsilateral ventricular dilatation, despite the presence of a mass lesion resembling a metastatic brain tumor. This interesting MR imaging finding is an important point for differentiating between a brain tumor and cerebral sparganosis.


What influences the standards of "quality" of carotid endarterectomy?
This paper analyzes the short-term results of 463 consecutive primary reconstructive operations on the carotid bifurcation based on the standards of "quality". In 383 (82.7%) cases, the patients were operated on for stenosis of the internal carotid artery (ICA); 60 (13%) patients had combination of stenosis and kingking of the ICA and 20 (4.3%) presented with isolated kinking of the ICA. Classic open carotid endarterectomy (CEAE) was performed in 240 cases, eversion CEAE in 144, resection with ICA redressment in 45, and ICA grafting in 28 cases. The perioperative lethality accounted for 1.54% (7 cases), per 463 operations, with the standard of "quality" being equal to 2%. Myocardial infarction was the cause of death in 5 cases, one patient developed homolateral stroke which eventuated in lethal outcome, and one more patient developed lethal stroke in the contralateral hemisphere. The incidence of homolateral strokes which did not eventuate in lethal outcome accounted for 1.94% (9 cases). The indicator "stroke plus lethality from stroke" in the general patient group was 2.38% which is even lower versus the standard of "quality" for asymptomatic patients and constitutes 3%. Among asymptomatic patients proper, the indicator "stroke plus lethality from stroke" was equal to 0, among TIA patients to 0 at a 5% standard, and in patients with initial stroke, initial stroke, it amounted to 3.19% at a 7% standard. Perioperative strokes were not recorded in patients with ICA kingking or at ICA grafting. The basic factor of the risk of perioperative neurologic deficit was a history of stroke. Patients with different stages of cerebrovascular insufficiency did not significantly differ as regards age, sex distribution, the presence of CAD, arterial hypertension, diabetes mellitus, the degree of stenosis of the operated ICA and the time of its clamping at operation. Patients with a history of strokes differed significantly from the general group only in the incidence of lesion of the contralateral ICA and in the frequency of the use of internal shunt. Analysis of the risk factors of perioperative stroke in patients with a history of strokes has shown that they were diabetes mellitus and the use of internal shunt. The development of perioperative stroke among patients with a history of stroke was not influenced by the incidence of contralateral ICA lesion, the standing of cerebral circulatory distress or the size of an ischemic focus. The initial character of ICA lesion and the type of reconstruction did not affect the origin of perioperative focal deficits or lethality.


Long-Distance Transport of Thiamine (Vitamin B1) Is Concomitant with That of Polyamines.
Thiamine (vitamin B1) is ubiquitous and essential for cell energy supply in all organisms as a vital metabolic cofactor, known for over a century. In plants, it is established that biosynthesis de novo is taking place predominantly in green tissues and is furthermore limited to plastids. Therefore, transport mechanisms are required to mediate the movement of this polar metabolite from source to sink tissue to activate key enzymes in cellular energy generating pathways but are currently unknown. Similar to thiamine, polyamines are an essential set of charged molecules required for diverse aspects of growth and development, the homeostasis of which necessitates long-distance transport processes that have remained elusive. Here, a yeast-based screen allowed us to identify Arabidopsis (Arabidopsis thaliana) PUT3 as a thiamine transporter. A combination of biochemical, physiological, and genetic approaches permitted us to show that PUT3 mediates phloem transport of both thiamine and polyamines. Loss of function of PUT3 demonstrated that the tissue distribution of these metabolites is altered with growth and developmental consequences. The pivotal role of PUT3 mediated thiamine and polyamine homeostasis in plants, and its importance for plant fitness is revealed through these findings.


Influence of roasted flaxseed marc flour on rheological, structural, fermentation, water distribution, and migration properties of wheat dough.
Roasted flaxseed (RF) marc, which is a by-product obtained from RF oil extraction, has high nutritional value. This study evaluated the impact of RF marc flour on rheological, structural, fermentation, water distribution, and migration properties of the wheat dough. Results showed that adding RF into wheat flour (WF) could effectively increase the water absorption in the dough and retard the retrogradation of starch. The fermentation results revealed that adding RF could improve the gas retention coefficient of dough. Compared to the dough prepared with WF, the doughs enriched with RF had higher tan δ values (ratio of loss modulus G″ to storage modulus G'), indicating a more liquid-like property. The mobility of tightly bound and free water in dough was decreased by adding RF, whereas the distribution of free water was increased. On the one hand, adding RF would dilute the gluten content in dough, resulting in a weaker protein network. On the other hand, the dietary fiber and proteins in RF could offset the gluten dilution effect to some extent. Overall, the results suggested that the substitution level of RF in WF should be below 25% to avoid serious dough quality deterioration, and the RF-WF blended flour could be a potential ingredient to produce wheat products with moist taste. These findings could be useful for guiding the future usage of RF marc in wheat-based products.


Alpha basic crystallin expression in canine mammary tumors.
The aim of this study was to evaluate prognostic and/or diagnostic factors of canine mammary tumors by immunohistochemically analyzing the expression of alpha basic crystallin (αB-c). For this, formalin-fixed, paraffin-embedded blocks of 51 naturally-occurring canine mammary tumors (11 benign and 40 malignant) were used. Tissue from eight normal canine mammary glands were served as a control. Immunohistochemically, in the control mammary tissues, a few luminal epithelial cells were αB-c positive but myoepithelial cells were negative. In benign or simple type malignant tumors, αB-c expression was observed in luminal epithelial cells while the myoepithelial basal cells were negative. In benign or complex type malign tumors, positive staining was predominantly found in the cytoplasm of epithelial cells. Immunoreactivity of αB-c was also observed in neoplastic myoepithelial cells. Statistically, the number of cells immunolabeled with αB-c was found to be significantly different among tissues from normal canine mammary glands, benign lesions, and malignant tumors (p < 0.05). αB-c immunoreactivity was higher in malignant tumors than the control mammary tissues (p < 0.001). Data obtained in the current study revealed a strong association between high expression levels of αB-c and primary mammary gland tumors in canines.


[Long-term follow-up of children with neurogenic bladder and artificial urinary sphincter].
Since 1986 an artificial urinary sphincter was implanted in 16 children (13 males and 3 females) 4 to 12 years old (median age 9) in order to solve their urinary incontinence. Mean age at implantation was 9 years. In 10 children (63%) only the implant procedure of artificial sphincter was performed without any other procedure associated (Group A), while in 6 children an augmentation cystoplasty simultaneously was performed (Group B). The results were analysed in both groups separately: 1. Group A: 10 patients. Two are functioning excellently after 10 and 11 years of follow up. Eight developed some type of troublesome: Five developed a deterioration of urinary tract and all of five were treated by augmentation cystoplasty; one of them is in a good condition after 11 years, another presented a fail of the device after 10 years, it has been changed, and in the others three the apparatus was removed by diverse causes. Two developed a mechanical fail of device: one was removed due familiar decision and the other has received another device. One urinary fistula developed 6 months later. The device was removed. 2. Group B: 6 patients (artificial sphincter and augmentation cystoplasty simultaneously). Five are functioning without trouble some. One persistent urinary fistula. The device was removed. Actually, of 16 cases (both groups) 8 cases (50%) are good functioning, 6 devices have been removed and 2 are waiting a new implant. The artificial urinary sphincter is a good solution for children with urinary incontinence in selected cases, but is mandatory a correct follow up because longterm complications can be developed. Results seem better when an augmentation cystoplasty is associated.


Heterogeneous amino acids in Ras and Rap1A specifying sensitivity to GAP proteins.
Guanosine triphosphatase (GTPase) activity of Ras is increased by interaction with Ras-GAP (GTPase-activating protein) or with the GAP-related domain of the type 1 neurofibromatosis protein (NF1-GRD), but Ras is not affected by interaction with cytoplasmic and membrane forms of Rap-GAP; Rap1A, whose effector function can suppress transformation by Ras, is sensitive to both forms of Rap-GAP and resistant to Ras-GAP and NF1-GRD. A series of chimeric proteins composed of portions of Ras and Rap were constructed; some were sensitive to Ras-GAP but resistant to NF1-GRD, and others were sensitive to cytoplasmic Rap-GAP but resistant to membrane Rap-GAP. Sensitivity of chimeras to Ras-GAP and cytoplasmic Rap-GAP was mediated by amino acids that are carboxyl-terminal to the effector region. Residues 61 to 65 of Ras conferred Ras-GAP sensitivity, but a larger number of Rap1A residues were required for sensitivity to cytoplasmic Rap-GAP. Chimeras carrying the Ras effector region that were sensitive only to Ras-GAP or only to cytoplasmic Rap-GAP transformed NIH 3T3 cells poorly. Thus, distinct amino acids of Ras and Rap1A mediate sensitivity to each of the proteins with GAP activity, and transforming potential of Ras and sensitivity of Ras to Ras-GAP are at least partially independent properties.


Developing a Practical Medical Educator Training Program: Meeting A Need in Faculty Development.
This article was migrated. The article was marked as recommended. Faculty who teach in medical schools typically do so because of their knowledge and expertise in their field, yet few receive training in best practices in teaching. Educator development programs that help faculty enhance their teaching skills while continuing to fulfill their existing professional responsibilities can help address this gap. Such programs may be developed and implemented locally by individuals within the institution. This guide is intended for individuals who are interested in developing educator training programs but who lack experience in program development. The article describes practical strategies for designing, implementing, and evaluating a collaborative program to teach skills and best practices in medical education. Key themes in program design, program implementation, and program evaluation and dissemination include appropriate goal setting, setting clear expectations, strong communication, and the benefits of diversity in collaboration. Educator training programs provide enhanced teaching skills and opportunities for career advancement for participants at all career stages, which in turn benefits the institution and the medical profession.


TARANIS interacts with VRILLE and PDP1 to modulate the circadian transcriptional feedback mechanism in Drosophila.
The molecular clock that generates daily rhythms of behavior and physiology consists of interlocked transcription-translation feedback loops. In Drosophila , the primary feedback loop involving the CLOCK-CYCLE transcriptional activators and the PERIOD-TIMELESS transcriptional repressors is interlocked with a secondary loop involving VRILLE (VRI) and PAR DOMAIN PROTEIN 1 (PDP1), a repressor and activator of Clock transcription, respectively. Whereas extensive studies have found numerous transcriptional, translational, and post-translational modulators of the primary loop, relatively little is known about the secondary loop. In this study, we demonstrate that TARANIS (TARA), a Drosophila homolog of the TRIP-Br/SERTAD family of transcriptional coregulators, functions with VRI and PDP1 to modulate the circadian period and rhythm strength in Drosophila . Overexpression of TARA lengthens the circadian period, whereas tara mutants exhibit reduced rhythmicity, a faster molecular clock, and reduced expression of the PDF neuropeptide. We find that TARA can form a physical complex with VRI and PDP1, enhancing their repressor and activator functions, respectively. The conserved SERTA domain of TARA is required to regulate the transcriptional activity of VRI and PDP1, and its deletion leads to reduced locomotor rhythmicity. Consistent with TARA's role in enhancing VRI and PDP1 activity, overexpressing tara has a similar effect on the circadian period and rhythm strength as simultaneously overexpressing vri and Pdp1 . Together, our results suggest that TARA modulates circadian behavior by enhancing the transcriptional activity of VRI and PDP1. Internal molecular clocks generating circadian rhythms of around 24 hours broadly impact behavior and physiology, and circadian dysfunction is associated with various neurological and metabolic diseases. The Drosophila circadian clock is a valuable model for understanding the molecular mechanisms underlying daily rhythms. In this study, we identify a conserved gene, taranis , as a novel regulator of the Drosophila molecular clock. We show that TARANIS modulates circadian behavior by physically interacting with and enhancing the transcriptional activity of clock proteins VRILLE and PDP1. Since mammalian homologs of VRILLE and PDP1 also function in the molecular clock, our results have implications for understanding the mammalian circadian clock.


Host genetic background influences the response to the opportunistic Pseudomonas aeruginosa infection altering cell-mediated immunity and bacterial replication.
Pseudomonas aeruginosa is a common cause of healthcare-associated infections including pneumonia, bloodstream, urinary tract, and surgical site infections. The clinical outcome of P. aeruginosa infections may be extremely variable among individuals at risk and patients affected by cystic fibrosis. However, risk factors for P. aeruginosa infection remain largely unknown. To identify and track the host factors influencing P. aeruginosa lung infections, inbred immunocompetent mouse strains were screened in a pneumonia model system. A/J, BALB/cJ, BALB/cAnNCrl, BALB/cByJ, C3H/HeOuJ, C57BL/6J, C57BL/6NCrl, DBA/2J, and 129S2/SvPasCRL mice were infected with P. aeruginosa clinical strain and monitored for body weight and mortality up to seven days. The most deviant survival phenotypes were observed for A/J, 129S2/SvPasCRL and DBA/2J showing high susceptibility while BALB/cAnNCrl and C3H/HeOuJ showing more resistance to P. aeruginosa infection. Next, one of the most susceptible and resistant mouse strains were characterized for their deviant clinical and immunological phenotype by scoring bacterial count, cell-mediated immunity, cytokines and chemokines profile and lung pathology in an early time course. Susceptible A/J mice showed significantly higher bacterial burden, higher cytokines and chemokines levels but lower leukocyte recruitment, particularly neutrophils, when compared to C3H/HeOuJ resistant mice. Pathologic scores showed lower inflammatory severity, reduced intraluminal and interstitial inflammation extent, bronchial and parenchymal involvement and diminished alveolar damage in the lungs of A/J when compared to C3H/HeOuJ. Our findings indicate that during an early phase of infection a prompt inflammatory response in the airways set the conditions for a non-permissive environment to P. aeruginosa replication and lock the spread to other organs. Host gene(s) may have a role in the reduction of cell-mediated immunity playing a critical role in the control of P. aeruginosa infection. These results now provide a basis for mapping genomic regions underlying host susceptibility to P. aeruginosa infection.


Effect of serum storage, anti-inflammatory oral doses of prednisone, and spontaneous hyperadrenocorticism on serum glutamate dehydrogenase activity in dogs.
Glutamate dehydrogenase (GLDH) is a mitochondrial enzyme with highest activity in periacinar hepatocytes. It is reported to be a sensitive indicator of hepatic injury; however, results of studies regarding tissue specificity are contradictory. The purpose of the study reported here was to examine the effect of 3 factors on serum GLDH activity in dogs: serum storage, anti-inflammatory oral doses of prednisone, and spontaneous hyperadrenocorticism (HAC). Stability of enzyme activity was determined by comparing serum samples stored at approximately 20 degrees C, 4 degrees C, and 20 degrees C for 4, 24, 48, and 72 hours, 1 week, and 6 months. To determine whether orally administered prednisone affected GLDH activity, the median difference in serum GLDH activity was compared between 5 untreated control dogs and 8 dogs that had received a tapering oral dose of prednisone. Lastly, GLDH enzyme activity was compared between 17 dogs with HAC and 16 age-matched controls. GLDH activity remained stable for 48 hours, 1 week, and 6 months, in serum stored at approximately 20 degrees C, 4 degrees C, and 20 degrees C, respectively. The median change in GLDH activity was not significantly different between dogs receiving prednisone and controls; however, dogs with HAC had significantly higher values than those of age-matched controls. Serum samples should be maintained at 4 degrees C if analysis of GLDH activity will be delayed by >48 hours; serum stored at 20 degrees C yields reliable results for up to 6 months. Serum GLDH activity was not increased in most dogs receiving short-term, anti-inflammatory oral doses of prednisone, in contrast to its increased activity in dogs with HAC.


The dopamine transporter constitutively internalizes and recycles in a protein kinase C-regulated manner in stably transfected PC12 cell lines.
The dopamine transporter (DAT) removes dopamine from the extracellular milieu and is potently inhibited by number of psychoactive drugs, including cocaine, amphetamines, and methylphenidate (Ritalin). Multiple lines of evidence demonstrate that protein kinase C (PKC) down-regulates dopamine transport, primarily by redistributing DAT from the plasma membrane to endosomal compartments, although the mechanisms facilitating transporter sequestration are not defined. Here, we demonstrate that DAT constitutively internalizes and recycles in rat pheochromocytoma (PC12) cells. Temperature blockades demonstrated basal internalization and reliance on recycling to maintain DAT cell surface levels. In contrast, recycling blockade with bafilomycin A1 significantly decreased transferrin receptor (TfR) surface expression but had no effect on DAT surface levels, suggesting that DAT and TfR traffic via distinct endosomal mechanisms. Kinetic analyses reveal robust constitutive DAT cycling to and from the plasma membrane, independent of transporter expression levels. In contrast, phorbol ester-mediated PKC activation accelerated DAT endocytosis and attenuated transporter recycling in a manner sensitive to DAT expression levels. These data demonstrate constitutive DAT trafficking and that PKC-mediated DAT sequestration is achieved by a combination of accelerated internalization and reduced recycling. Additionally, the differential sensitivity to expression level exhibited by constitutive and regulated DAT trafficking suggests that these two processes are mediated by independent cellular mechanisms.


A biology-driven approach identifies the hypoxia gene signature as a predictor of the outcome of neuroblastoma patients.
Hypoxia is a condition of low oxygen tension occurring in the tumor microenvironment and it is related to poor prognosis in human cancer. To examine the relationship between hypoxia and neuroblastoma, we generated and tested an in vitro derived hypoxia gene signature for its ability to predict patients' outcome. We obtained the gene expression profile of 11 hypoxic neuroblastoma cell lines and we derived a robust 62 probesets signature (NB-hypo) taking advantage of the strong discriminating power of the l1-l2 feature selection technique combined with the analysis of differential gene expression. We profiled gene expression of the tumors of 88 neuroblastoma patients and divided them according to the NB-hypo expression values by K-means clustering. The NB-hypo successfully stratifies the neuroblastoma patients into good and poor prognosis groups. Multivariate Cox analysis revealed that the NB-hypo is a significant independent predictor after controlling for commonly used risk factors including the amplification of MYCN oncogene. NB-hypo increases the resolution of the MYCN stratification by dividing patients with MYCN not amplified tumors in good and poor outcome suggesting that hypoxia is associated with the aggressiveness of neuroblastoma tumor independently from MYCN amplification. Our results demonstrate that the NB-hypo is a novel and independent prognostic factor for neuroblastoma and support the view that hypoxia is negatively correlated with tumors' outcome. We show the power of the biology-driven approach in defining hypoxia as a critical molecular program in neuroblastoma and the potential for improvement in the current criteria for risk stratification.


Solid cancers after bone marrow transplantation.
To evaluate the incidence and associated risk factors of solid cancers after bone marrow transplantation (BMT). We analyzed 2,129 patients who had undergone BMT for hematologic malignancies at the City of Hope National Medical Center between 1976 and 1998. A retrospective cohort and nested case-control study design were used to evaluate the role of pretransplantation therapeutic exposures and transplant conditioning regimens. Twenty-nine patients developed solid cancers after BMT, which represents a two-fold increase in risk compared with a comparable normal population. The estimated cumulative probability (+/- SE) for development of a solid cancer was 6.1% +/- 1.6% at 10 years. The risk was significantly elevated for liver cancer (standardized incidence ratio [SIR], 27.7; 95% confidence interval [CI], 1.9 to 57.3), cancer of the oral cavity (SIR, 17.4; 95% CI, 6.3 to 34.1), and cervical cancer (SIR, 13.3; 95% CI, 3.5 to 29.6). Each of the two patients with liver cancer had a history of chronic hepatitis C infection. All six patients with squamous cell carcinoma of the skin had chronic graft-versus-host disease. The risk was significantly higher for survivors who were younger than 34 years of age at time of BMT (SIR, 5.3; 95% CI, 2.7 to 8.6). Cancers of the thyroid gland, liver, and oral cavity occurred primarily among patients who received total-body irradiation. The risk of radiation-associated solid tumor development after BMT is likely to increase with longer follow-up. This underscores the importance of close monitoring of patients who undergo BMT.


OpenDrop: An Integrated Do-It-Yourself Platform for Personal Use of Biochips.
Biochips, or digital labs-on-chip, are developed with the purpose of being used by laboratory technicians or biologists in laboratories or clinics. In this article, we expand this vision with the goal of enabling everyone, regardless of their expertise, to use biochips for their own personal purposes. We developed OpenDrop, an integrated electromicrofluidic platform that allows users to develop and program their own bio-applications. We address the main challenges that users may encounter: accessibility, bio-protocol design and interaction with microfluidics. OpenDrop consists of a do-it-yourself biochip, an automated software tool with visual interface and a detailed technique for at-home operations of microfluidics. We report on two years of use of OpenDrop, released as an open-source platform. Our platform attracted a highly diverse user base with participants originating from maker communities, academia and industry. Our findings show that 47% of attempts to replicate OpenDrop were successful, the main challenge remaining the assembly of the device. In terms of usability, the users managed to operate their platforms at home and are working on designing their own bio-applications. Our work provides a step towards a future in which everyone will be able to create microfluidic devices for their personal applications, thereby democratizing parts of health care.