Self-assembled IR780-loaded transferrin nanoparticles as an imaging, targeting and PDT/PTT agent for cancer therapy.
Combination of photothermal and photodynamic therapy (PTT/PDT) offer unique advantages over PDT alone. However, to achieve synergetic PDT/PTT effect, one generally needs two lasers with different wavelengths. Near-infrared dye IR-780 could be used as photosensitizer both for PTT and PDT, but its lipophilicity limits its practical use and in vivo efficiency. Herein, a simple multifunctional IR780-loaded nanoplatform based on transferrin was developed for targeted imaging and phototherapy of cancer compatible with a single-NIR-laser irradiation. The self-assembled transferrin-IR780 nanoparticles (Tf-IR780 NPs) exhibited narrow size distribution, good photo-stability, and encouraging photothermal performance with enhanced generation of ROS under laser irradiation. Following intravenous injection, Tf-IR780 NPs had a high tumor-to-background ratio in CT26 tumor-bearing mice. Treatment with Tf-IR780 NPs resulted in significant tumor suppression. Overall, the Tf-IR780 NPs show notable targeting and theranostic potential in cancer therapy.


A good outcome following complete injury of the brachial plexus: long-term analysis of the management of two patients.
Restoration of hand function is rarely achieved after a complete closed traction lesion of the supraclavicular brachial plexus. We describe the injury, treatment, rehabilitation and long-term results of two patients who regained good function of the upper limb and useful function in the hand after such an injury. Successful repairs were performed within six days of injury. Tinel's sign proved accurate in predicting the ruptures and the distribution of pain was accurate in predicting avulsion. The severe pain that began on the day of injury resolved with the onset of muscle function. Recovery of muscle function preceded recovery of sensation. Recovery of the function of C and Adelta fibres was the slowest of all.


Prevention of early postnatal hyperalimentation protects against activation of transforming growth factor-β/bone morphogenetic protein and interleukin-6 signaling in rat lungs after intrauterine growth restriction.
Intrauterine growth restriction (IUGR) is intimately linked with postnatal catch-up growth, leading to impaired lung structure and function. However, the impact of catch-up growth induced by early postnatal hyperalimentation (HA) on the lung has not been addressed to date. The aim of this study was to investigate whether prevention of HA subsequent to IUGR protects the lung from 1) deregulation of the transforming growth factor-β(TGF-β)/bone morphogenetic protein (BMP) pathway, 2) activation of interleukin (IL)-6 signaling, and 3) profibrotic processes. IUGR was induced in Wistar rats by isocaloric protein restriction during gestation by feeding a control (Co) or a low-protein diet with 17% or 8% casein, respectively. On postnatal day 1 (P1), litters from both groups were randomly reduced to 6 pups per dam to induce HA or adjusted to 10 pups and fed with standard diet: Co, Co with HA (Co-HA), IUGR, and IUGR with HA (IUGR-HA). Birth weights in rats after IUGR were lower than in Co rats (P < 0.05). HA during lactation led to accelerated body weight gain from P1 to P23 (Co vs. Co-HA, IUGR vs. IUGR-HA; P < 0.05). At P70, prevention of HA after IUGR protected against the following: 1) activation of both TGF-β [phosphorylated SMAD (pSMAD) 2; plasminogen activator inhibitor 1 (Pai1)] and BMP signaling [pSMAD1; inhibitor of differentiation (Id1)] compared with Co (P < 0.05) and Co or IUGR (P < 0.05) rats, respectively; 2) greater mRNA expression of interleukin (Il) 6 and Il13 (P < 0.05) as well as activation of signal transducer and activator of transcription 3 (STAT3) signaling (P < 0.05) after IUGR-HA; and 3) greater gene expression of collagen Iα1 and osteopontin (P < 0.05) and increased deposition of bronchial subepithelial connective tissue in IUGR-HA compared with Co and IUGR rats. Moreover, HA had a significant additive effect (P < 0.05) on the increased enhanced pause (indicator of airway resistance) in the IUGR group (P < 0.05) at P70. This study demonstrates a dual mechanism in IUGR-associated lung disease that is 1) IUGR-dependent and 2) HA-mediated and thereby offers new avenues to develop innovative preventive strategies for perinatal programming of adult lung diseases.


Classification of Brain Tumor Images Using CNN.
A brain tumor is a serious malignant condition caused by unregulated as well as aberrant cell partitioning. Recent advances in deep learning have aided the healthcare business, particularly, diagnostic imaging for the diagnosis of numerous disorders. The most frequent and widely utilized machine learning model for image recognition is probably task CNN. Similarly, in our study, we categorize brain MRI scanning images using CNN and data augmentation and image processing techniques. We compared the performance of the scratch CNN model with that of pretrained VGG-16 models using transfer learning. Even though the investigation is carried out on a small dataset, the results indicate that our model's accuracy is quite successful and has extremely low complexity rates, achieving 100 percent accuracy compared to 96 percent accuracy for VGG-16. Compared to existing pretrained methods, our model uses much less processing resources and produces substantially greater accuracy.


Tubulointerstitial nephritis and vasculitis.
This review discusses recent developments in infectious tubulointerstitial nephritis, including bacterial virulence factors, the interaction between leukocytes and bacteria, and early events in inflammation. In the area of vasculitis, little-studied specificities of antineutrophil cytoplasmic antibodies (ANCAs), including lactoferrin and elastase, plus newly described ANCAs with alpha-enolase specificity are discussed, along with newer studies on the pathogenesis of ANCA-mediated damage and animal models for ANCA-related vasculitis.


Evaluation of the level of domestic hygiene in household kitchens.
Each year in industrialized countries, 30-40% or more cases of food poisoning occur in the household. The aims of this study are to describe the aspects related to food safety in households by carrying out a microbiological characterization of the kitchens in residential dwellings and to increase consumers' awareness concerning the importance of good hygienic practices, which are required for preventing foodborne diseases at household level. The collection of data involved taking 760 analytical samples of kitchen-counter tops and food from 80 kitchens of private dwellings; the manner in which food was treated was evaluated from the moment of purchase to its transformation and storage by means of a checklist; the questionnaire enabled us to determine the extent of consumers' knowledge concerning the food sector. Overall, the results showed a good level of hygiene concerning both food and kitchen-counter top food surfaces. The respondents of the questionnaire did not appear to be aware of various health risks especially concerning the management of the temperature and compartments of refrigerators as well as food storage times and the cooking or heating of food. The data indicate that final consumers should take more care when cleaning kitchen-counter tops and washing salad; special training programmes should be included in school curricular in order to increase citizens' awareness and knowledge concerning food risks within the household.


[Study on the efficiency of the European System for Cardiac Operative Risk Evaluation (EuroSCORE) in predicting in-hospital mortality of the patients with percutaneous coronary intervention].
To investigate the efficiency of European System for Cardiac Operative Risk Evaluation (EuroSCORE) in predicting in-hospital mortality for the patients after percutaneous coronary intervention (PCI). Retrospective analysis was conducted on the patients who had undergone PCI in our hospital since year 2005 to 2007. We used both cumulative EuroSCORE score and logistic EuroSCORE to predict the in-hospital morality and to analyze the correlation between the predicted mortality and the actual mortality. According to the additive EuroSCORE, we divided the patients into three groups, the additive EuroSCORE 0-2 were divided into low-risk group, 3-5 were divided into mid-risk group and ≥ 6 into high-risk group. The actual in-hospital mortality rates were 0%, 0.47% and 6.09% respectively. The EuroSCORE model demonstrated an overall relation between the EuroSCORE ranking and the incidence of in-hospital mortality (P<0.001). Results from the multivariable logistic regression analysis showed that the EuroSCORE was an independent in-hospital mortality predictor (P<0.01). The EuroSCORE risk model and the in-hospital mortality were significantly correlated, indicating that the model was a promising method for predicting the in-hospital mortality of PCI patients.


Microsatellite allele A5.1 of MHC class I chain-related gene A is associated with latent autoimmune diabetes in adults in Latvia.
NIDDM is one of the most common forms of diabetes. The diagnosis is based on WHO classification, which is a clinical classification and misses the autoimmune diabetes in adults. Therefore, among the clinically diagnosed NIDDM cases, there can be a certain number of patients with latent autoimmune diabetes in adults (LADA). The MICA gene is located in the MHC class I region and is expressed by monocytes, keratinocytes, and endothelial cells. Sequence determination of the MICA gene identifies trinucleotide repeat (GCT) microsatellite polymorphism, which identifies 5 alleles with 4, 5, 6, and 9 repetitions of GCT (A4, A5, A6, and A9) or 5 repetitions of GCT with 1 additional G insertion for allele A5.1. From our previous studies, we have shown that microsatellite allele A5 of MICA is associated with IDDM. The aim of this study was to test the hypothesis that certain MICA alleles are associated with LADA among clinically diagnosed NIDDM. Out of 100 clinically diagnosed NIDDM patients, 49 tested positive for GAD65 and IA-2 antibodies by use of 35S RIA. Samples from these 49 patients and 96 healthy controls were analyzed for MICA by PCR amplification, and fragment sizes were determined in an ABI prism DNA sequencer. Our results show that MICA allele A5.1 is significantly increased in antibody-positive (GAD65 or IA-2) NIDDM patients [35/49 (72%)] when compared to healthy controls [22/96 (23%)] (OR = 8.4; P < 0.0001). However, we do not see any association with each of the antibodies separately. From our study, we conclude that (a) MICA allele A5.1 is associated with LADA and (b) MICA may play an important role in the etiopathogenesis of LADA.


Prolonged cardiopulmonary resuscitation during acute disopyramide poisoning.
A case is reported of sustained electromechanical dissociation due to deliberate disopyramide over-dosage in a young healthy girl. Circulatory recovery was achieved with standard resuscitative measures, including prolonged cardiac massage and isoproterenol infusion plus high dose calcium chloride. Neurological recovery was complete.


Genome Engineering and Modification Toward Synthetic Biology for the Production of Antibiotics.
Antibiotic production is often governed by large gene clusters composed of genes related to antibiotic scaffold synthesis, tailoring, regulation, and resistance. With the expansion of genome sequencing, a considerable number of antibiotic gene clusters has been isolated and characterized. The emerging genome engineering techniques make it possible towards more efficient engineering of antibiotics. In addition to genomic editing, multiple synthetic biology approaches have been developed for the exploration and improvement of antibiotic natural products. Here, we review the progress in the development of these genome editing techniques used to engineer new antibiotics, focusing on three aspects of genome engineering: direct cloning of large genomic fragments, genome engineering of gene clusters, and regulation of gene cluster expression. This review will not only summarize the current uses of genomic engineering techniques for cloning and assembly of antibiotic gene clusters or for altering antibiotic synthetic pathways but will also provide perspectives on the future directions of rebuilding biological systems for the design of novel antibiotics.


[Blood gastrin and colorectal neoplasms].
Gastrin has a trophic effect on the mucosa of the gastrointestinal tract and seems to have the potential for promoting colonic cancerogenesis through a chronic stimulation of the epithelial proliferation. Plasma gastrin has been reported to be elevated in patients with colorectal neoplasms. The aim of the present study was to verify this observation. Presurgical serum levels of gastrin were compared between 49 patients with colorectal neoplasms and 47 controls hospitalized for other surgical lesions. Results show significantly higher gastrin levels of case group than controls: 72.72 + 85.41 vs. 46.79 + 24.09 pg/ml (p < 0.05), and provide support for the hypothesis of a gastrin-stimulated neoplastic growth enhancing at the same time the potential therapeutic role of reducing gastrin secretion.


A MONTE CARLO STUDY OF SIMULATED MEASUREMENTS OF RADIONUCLIDES IN BONE.
When measuring the internally deposited activity in the bone of a subject, the placement of the detector is critical. This study reports the simulated counting efficiencies for three counting geometries, the skull, knee and shin, using 13 different voxel phantoms. It shows that the range of counting efficiencies for a given geometry is large for the studied phantoms, especially at low energies. Skull counting offers higher efficiency for low energies such as the 17 keV compared to knee counting or shin counting, but this advantage disappears when the energy is higher such as at 185 keV. This work also shows that the calibration phantom may greatly impact the accuracy of the activity estimate in bone counting, with uncertainties increasing greatly as the photon energy is reduced. Estimating the activity of a radionuclide in bone from direct counting has large uncertainties, and the dose calculated from a skeleton measurement would need careful analysis and, if possible, supporting data from other bioassay measurements.


Low-Intensity Pulsed Ultrasound Stimulation Enhances Heat-Shock Protein 90 and Mineralized Nodule Formation in Mouse Calvaria-Derived Osteoblasts.
Low-intensity pulsed ultrasound (LIPUS) has demonstrated its positive effects on osteogenic differentiation of mesenchymal stem cells and the proliferation and differentiation of osteoblasts, negative effects on osteoclast growth, and promotion of angiogenesis, leading to improvement of the tissue perfusion. Heat-shock proteins (HSPs) are initially identified as molecules encouraged and expressed by heat stress or chemical stress to cells and involved in the balance between differentiation and apoptosis of osteoblasts. However, it remains unclear if the effect of LIPUS on osteoblast differentiation could involve HSP expression and contribution. In this study, mouse calvarial osteoblasts were exposed to LIPUS at a frequency of 3.0 MHz by 30 mW/cm(2) for 15 min or to 42°C heat shock for 20 min at day 3 of cell culture and examined for osteogenesis with pursuing induction of HSP27, HSP70, and HSP90. LIPUS as well as heat shock initially upregulated HSP90 and phosphorylation of Smad1 and Smad5, encouraging cell viability and proliferation at 24 h, enhancing mineralized nodule formation stronger by LIPUS after 10 days. However, HSP27, associated with BMP2-stimulated p38 mitogen-activated protein kinase during osteoblast differentiation, was downregulated by both stimulations at this early time point. Notably, these two stimuli maintained Smad1 phosphorylation with mineralized nodule formation even under BMP2 signal blockage. Therefore, LIPUS might be a novel inducer of osteoblastic differentiation through a noncanonical signal pathway. In conclusion, LIPUS stimulation enhanced cell viability and proliferation as early as 24 h after treatment, and HSP90 was upregulated, leading to dense mineralization in the osteoblast cell culture after 10 days.


Pancreatic cancer detected by positron emission tomography with 18F-labelled deoxyglucose: method and first results.
In order to evaluate the utility of positron emission tomography (PET) with 18F-labelled deoxyglucose (FDG) for detection of pancreatic cancer 15 patients with pancreatic masses shown by computed tomography were investigated. Static PET scans covering an axial field of view of 15 cm were obtained 45 min after intravenous injection of 150-300 MBq FDG. Focally increased FDG accumulation was present in 12 out of 13 patients with histologically proven adenocarcinoma, in particular in eight of nine lymph node and four of five liver metastases. Scans of two patients with chronic pancreatitis confirmed by surgery revealed a normal FDG distribution. Contrast between tumour and normal tissue depended the metabolic situation prior to FDG injection. High ratios were found in fasting patients whereas no elevated FDG uptake was measured in an insulin-dependent diabetic suffering from carcinoma of the pancreatic head. We conclude that FDG PET might have the potential for detection and even differentiation of pancreatic carcinoma from chronic pancreatitis. Further studies are necessary to substantiate these preliminary findings and to optimize results in diabetic patients.


Endothelial microparticles correlate with high-risk angiographic lesions in acute coronary syndromes.
Endothelial Microparticles (EMP) are small fragments of endothelial cell membrane shed during apoptosis or activation. Our group has previously reported elevations of EMP in patients with coronary artery disease (CAD), thrombotic thrombocytopenic purpura (TTP), pre-eclampsia, multiple sclerosis (MS), and severe hypertension (HTN). In the present study, we evaluate the possible relationship between EMP levels and the angiographic severity and characteristics of coronary obstructive lesions. We studied a total of 43 patients undergoing coronary angiography. Fifteen had presented with acute myocardial infarction (MI), 20 with unstable anginas (UA), 5 with stable angina (SA) and 3 with congestive heart failure. Coronary angiography was reviewed and coronary lesions were classified using the Ambrose classification. Coronary stenoses were classified as high and low risk. High-risk included lesions with eccentric appearance (type II), presence of thrombi, or multiple irregularities. Low-risk lesions were defined as concentric or type I. Lesions were also analyzed by degree of stenosis and history of acute coronary syndrome (ACS). EMP in plasma was assayed by flow cytometry. EMP in eccentric type II or multiple irregular lesions (high-risk) were 2.5-fold higher than in type I or concentric (low-risk) lesions, p<0.05. Lesions with thrombi had three-fold higher EMP than those without (p=0.05). Mild stenosis (>20%-<45%) had three-fold higher EMP than more severe (>45%), and five-fold higher than those without stenosis (p<0.01). Among patients with type II lesions, those with first ACS episode had four-fold higher EMP levels than those with recurrent ACS (p<0.01). High EMP was associated with high-risk angiographic lesions including eccentric type II, multiple irregular, and lesions with thrombi. Mild to moderate stenosis was associated with higher EMP levels than severe stenosis. EMP may be a useful marker in detecting endothelial injury and risk of ACS as defined by angiography.


Men's experience of their partners' breast cancer diagnosis, breast surgery and oncological treatment.
To investigate the experiences of male partners of female breast cancer patients who had undergone surgery and oncological treatment and who were still raising children. Research on the psychological effects of breast cancer has focused primarily on the patients undergoing treatment, neglecting the effect of such a condition on their closest family members. This study addresses this gap by focusing on understanding the effects of this disease on male partners of these patients. An interpretative phenomenological approach was used. Eight males whose female partners were diagnosed with primary breast cancer between the ages of 30 and 55 and who had young children still living at home at the time were interviewed by the first author of this article. Interviews were transcribed verbatim and analyzed using an interpretative phenomenological framework, in accordance with the guidelines in the COREQ checklist for qualitative studies. Participants emphasized the difficulties they faced in trying to juggle work and family responsibilities while offering support to their partners, a task they felt ill-prepared for. At times, they felt excluded by their partners and worried about the impact of the disease on their children. A common fear was that of recurrence of the disease, and while some discussed the financial difficulties associated with treating the disease, others saw it as enhancing the potential for their personal and couple growth. This study adds to the topic by uncovering the perspectives of male partners of breast cancer patients and the effects of their partner's condition on them and their families. The results of this study can be used to inform policy when providing holistic care. They also highlight the importance of counselling and support interventions for partners of breast cancer patients.


Data on the effect of sex on the size, cellular content, and neuronal density of the developing brain in mice exposed to isoflurane and carbon monoxide.
The data presented here detail the changes in size, cellular content, and neuronal density of the developing brain over time with respect to sex in C57Bl/6 mice following neonatal exposure to isoflurane, carbon monoxide, or their combination. Specifically, brain weight- and brain volume-to-body weight ratios are presented, representative immunoblots of whole brain cell-specific protein content are depicted, and quantification of the number of neurons in the primary somatosensory cortex and CA3 region of the hippocampus are shown. Three discrete postnatal time points are represented: P7 (prior to exposure), P14 (one-week post exposure), and P42-56 (5-7 weeks post exposure). Major findings from the data presented here are reported in the manuscript "Carbon Monoxide Incompletely Prevents Isoflurane-induced Defects in Murine Neurodevelopment" (Wang et al., in press) [1].


Impact of fatigue at the shoulder on the contralateral upper limb kinematics and performance.
Altered movement patterns have been proposed as an etiological factor for the development of musculoskeletal pain. Fatigue influences upper limb kinematics and movement performance which could extend to the contralateral limb and potentially increasing risk of injury. The aim of this study was to investigate the impact of fatigue at the dominant arm on the contralateral upper limb movement. Forty participants were randomly assigned to one of two groups: Control or Fatigue Group. All participants completed a reaching task at the baseline and post-experimental phase, during which they reached four targets with their non-dominant arm in a virtual reality environment. Following the baseline phase, the Fatigue Group completed a shoulder fatigue protocol with their dominant arm only, while the Control Group took a 10-minute break. Thereafter, the reaching task was repeated. Upper limb and trunk kinematics (joint angles and excursions), spatiotemporal (speed and accuracy) and surface electromyographic (sEMG) activity (sEMG signal mean epoch amplitude and median frequency of the EMG power spectrum) were collected. Two-way repeated-measures ANOVA were performed to determine the effects of Time, Group and of the interaction between these factors. There was a significant Time x Group interaction for sternoclavicular elevation range of motion (p = 0.040), movement speed (p = 0.043) and accuracy (p = 0.033). The Fatigue group showed higher contralateral sternoclavicular elevation and increased movement error while experiencing fatigue in the dominant arm. Moreover, the Control group increased their speed during the Post-experimental phase compared to baseline (p = 0.043), while the Fatigue group did not show any speed improvement. There was no EMG sign of fatigue in any of the muscles evaluated. This study showed that fatigue at the dominant shoulder impacts movement at the contralateral upper limb. Such changes may be a risk factor for the development of shoulder pain in both the fatigued and non-fatigued limbs.


Central nervous system: recovery of function.
Rats with one- or two-stage lesions in the hippocampus, anzygdala, or frontal cortex were tested on a variety of tasks. Even though locus and amount of tissue damage were very similar in the two groups, animals with one-stage lesions were impaired as judged by performance on all problems, while aninmals with two-stage lesionis were not.


Cytochrome P450 inactivation during reductive metabolism of 1,1-dichloro-2,2,2-trifluoroethane (HCFC-123) by phenobarbital- and pyridine-induced rat liver microsomes.
The reductive metabolic activation of 1,1-dichloro-2,2,2-trifluoroethane (HCFC-123), one of the potential substitutes for the ozone-depleting chlorofluorocarbons and a close structural analogue of the hepatotoxic anesthetic halothane, was investigated in vitro. During incubation of liver microsomes from phenobarbital-(PB) or pyridine-induced (PYR) rats with 0-20 mM HCFC-123 under anaerobic conditions, a dose- and time-dependent depletion of added exogenous glutathione was observed, indicating the formation of reactive metabolites. Under similar incubation conditions, except for the absence of glutathione, 1-chloro-2,2,2-trifluoroethane and 1-chloro-2,2-difluoroethene were detected as products of reductive metabolism of HCFC-123, as previously reported for halothane. As shown previously in our laboratory for halothane, under these conditions HCFC- 123 also caused a statistically significant loss of microsomal cytochrome P450 (P450) as indicated by a decrease of the classical absorption spectrum in the presence of CO. Both glutathione depletion and P450 loss were almost completely prevented by previous saturation of the incubation mixture with CO and were partially prevented by the presence of the free-radical scavenger N-t-butyl-alpha-phenylnitrone or the carbene trapping agent 2,3-dimethyl-2-butene, suggesting that both types of intermediates may be involved. The loss of P450 was associated with a quantitatively similar loss of microsomal heme, as measured by the pyridine hemochromogen reaction, with PB but not with PYR microsomes. Finally, both the P4502E1-specific p-nitrophenol hydroxylase activity in PYR microsomes and the P4502B1/2-specific pentoxyresorufin O-depentylase activity in PB microsomes were significantly inhibited (58 and 53%, respectively) by prior incubation with HCFC-123, suggesting that both isoforms are able to catalyze the activation of this halogenated compound. These results indicate that indeed HCFC-123, like its analogue halothane, is activated reductively to reactive metabolites by at least two P450 isoforms, namely P4502E1 and P4502B1/2. These metabolites, probably free radicals and/or carbene species, may attack the enzyme resulting in modification of the heme group and subsequent loss of catalytic activity.


Intensive care management of neonates with d-transposition of the great arteries and common arterial trunk.
Although mortality rates for patients with d-transposition of the great arteries remain quite low, these patients have a unique circulation that requires careful management in the peri-operative period. Despite the improved mortality for patients with common arterial trunk, the course in the intensive care unit is remarkable for significant morbidity and utilisation of significant resources. Pre-operative patient management focuses on balancing competing circulations, pulmonary and systemic, which exist in parallel rather than in series, as in the normal circulation. Post-operative patient management in both lesions focuses on optimising systemic output, respiratory status, and mitigating the effects of cardiopulmonary bypass. In this article, we review pre- and post-operative intensive care management in neonates with d-transposition of the great arteries and common arterial trunk.


Extended hours hub is a help.
I was interested to read that doctors had called for GP practices to introduce a 'black alert' system that could divert patients from overstretched surgeries (news, 5 July).


Resurrection of the earthworm species Dendrobaena fedtschenkoi (Michaelsen, 1900), a former synonym of Dendrobaena byblica (Rosa, 1893) (Clitellata: Megadrili).
Dendrobaena byblica (Rosa, 1893) is a Circum-Mediterranean earthworm species complex containing more than a dozen synonymous names. From these, only two species have been resurrected so far, D. ganglbaueri (Rosa, 1894) and Fitzingeria annectens (Rosa, 1895). Here we demonstrate that Dendrobaena fedtschenkoi (Michaelsen, 1900) described from the Pamir Mts. Tajikistan differs from D. byblica both in morphological and molecular features, and represents an independent species.


[Epidemic outbreak of meningitis due to Echovirus type 13 on the island of Gran Canaria (Spain)].
Since the implementation of routine vaccination against mumps, enteroviruses have become the commonest cause of viral meningitis. In Spain the most frequently isolated echoviruses are serotypes 30, 9, 6 and 4.Objectives. Determine the clinical-epidemiological characteristics of a meningitis outbreak due to echovirus type 13. We studied 152 cases of enteroviral meningitis diagnosed in the year 2000. Viral isolation was performed on fibroblast MRC-5 and human rhabdomyosarcoma cells. The viruses were identified with monoclonal antibodies and serotyped by neutralization. Echovirus was isolated in 131 out of 152 CSF samples (86.2%). In 21 patients with negative CSF culture, the diagnosis was made by echovirus isolation from upper respiratory tract and/or feces samples. The cytopathic effect was seen in all cases in monolayers of rhabdomyosarcoma cells. The average age of the patients was 67 months (range: 1-350) and the male:female ratio was 2:1. Most of the patients had fever, headache and other signs of meningitis. Hospitalization was required in 52.6%. All patients were discharged without sequelae. The highest incidence was recorded from April to June. There had been no record of the presence of echovirus 13 in Spain in recent years until the emergence of this outbreak. Isolation of the virus from sites other than CSF is often helpful in establishing the diagnosis and patient management. Enterovirus isolates must be specifically serotyped to determine enteroviral activity in the population, which can be sporadic or epidemic.


Dose- and schedule-dependent activation and drug synergism between thymidine and 5-aza-2'-deoxycytidine in a human promyelocytic leukemia cell line.
The ability of thymidine (dThd) to enhance the metabolism and cytotoxicity of subsequent administered 5-aza-2'-deoxycytidine (5-aza-dCyd) was studied in L1210 cells and in the human promyelocytic leukemic cell line, HL-60. Exposure of L1210 cells to 0.1 mM dThd for 5 h resulted in an increase in the total intracellular and acid-precipitable accumulation of 5-aza-dCyd. Higher dThd concentrations and longer exposure intervals resulted in smaller increments in 5-aza-dCyd accumulation. In contrast, in HL-60 cells, a 24-hr exposure in 1 mM dThd resulted in the greatest intracellular accumulation of 5-aza-dCyd, 3.3 times more accumulation than in control cells. There was also a 4-fold increase in the acid-precipitable accumulation and nearly a 3-fold increase in DNA incorporation of 5-aza-dCyd in HL-60 cells exposed to the same dThd schedule. High-pressure liquid chromatographic analysis demonstrated a greater than 3-fold increase in the intracellular amounts of 5-aza-dCyd metabolites eluting in the triphosphate region in these human cells under identical conditions. Shorter dThd incubation exposure intervals (6 hr) and lower dThd concentration (0.1 mM) produced smaller increments in these studies. Both growth and clonogenic assays of HL-60 cells demonstrated a dose- and schedule sequence-dependent synergism between dThd and 5-aza-dCyd.


Jasmonate-mediated defence responses, unlike salicylate-mediated responses, are involved in the recovery of grapevine from bois noir disease.
Bois noir is an important disease of grapevine (Vitis vinifera L.), caused by phytoplasmas. An interesting, yet elusive aspect of the bois noir disease is "recovery", i.e., the spontaneous and unpredictable remission of symptoms and damage. Because conventional pest management is ineffective against bois noir, deciphering the molecular bases of recovery is beneficial. The present study aimed to understand whether salicylate- and jasmonate-defence pathways might have a role in the recovery from the bois noir disease of grapevine. Leaves from healthy, bois noir-diseased and bois noir-recovered plants were compared, both in the presence (late summer) and absence (late spring) of bois noir symptoms on the diseased plants. Analyses of salicylate and jasmonate contents, as well as the expression of genes involved in their biosynthesis, signalling and action, were evaluated. In symptomatic diseased plants (late summer), unlike symptomless plants (late spring), salicylate biosynthesis was increased and salicylate-responsive genes were activated. In contrast, jasmonate biosynthesis and signalling genes were up-regulated both in recovered and diseased plants at all sampling dates. The activation of salicylate signalling in symptomatic plants might have antagonised the jasmonate-mediated defence response by suppressing the expression of jasmonate-responsive genes. Our results suggest that grapevine reacts to phytoplasma infection through salicylate-mediated signalling, although the resultant full activation of a salicylate-mediated response is apparently ineffective in conferring resistance against bois noir disease. Activation of the salicylate signalling pathway that is associated with the presence of bois noir phytoplasma seems to antagonise the jasmonate defence response, by failing to activate or suppressing both the expression of some jasmonate responsive genes that act downstream of the jasmonate biosynthetic pathway, as well as the first events of the jasmonate signalling pathway. On the other hand, activation of the entire jasmonate signalling pathway in recovered plants suggests the potential importance of jasmonate-regulated defences in preventing bois noir phytoplasma infections and the subsequent development of bois noir disease. Thus, on one hand, recovery could be achieved and maintained over time by preventing the activation of defence genes associated with salicylate signalling, and on the other hand, by activating jasmonate signalling and other defence responses.


COVID-19: Understanding and mitigating trauma in ICU survivors.
The spread of coronavirus disease 2019 (COVID-19) has placed many individuals in need of critical care, with a high proportion of hospitalized patients being admitted to intensive care units (ICU) to treat acute outcomes of COVID-19 (e.g., respiratory failure via mechanical ventilation). The ICU is known to be a setting where individuals are at a high risk of experiencing significant psychological difficulties, and patients with COVID-19 are particularly susceptible to such experiences, which can impact their recovery process (e.g., postintensive care syndrome). This article seeks to highlight the intersection between critical care related to trauma and COVID-19 and point providers toward opportunities for anticipating and managing secondary effects in effort to promote psychological adaptation. (PsycInfo Database Record (c) 2020 APA, all rights reserved).


Abundance of active and inactive microcystin genotypes in populations of the toxic cyanobacterium Planktothrix spp.
To investigate the abundance of active and inactive microcystin genotypes in populations of the filamentous cyanobacterium Planktothrix spp., individual filaments were grown as clonal strains in the laboratory and analysed for microcystin synthetase (mcy) genes and microcystin. Twenty-three green-pigmented strains of P. agardhii originating mostly from shallow water bodies fell into two groups, those possessing mcyA and those lacking mcyA. In contrast, all of the 49 strains that were assigned to the red-pigmented P. rubescens contained mcyA. One strain of P. agardhii and eight strains of P. rubescens contained the total microcystin synthetase gene cluster but were found inactive in microcystin synthesis. To investigate the natural abundance of inactive mcy genotypes in P. rubescens individual filaments sampled from Lake Irrsee and Lake Mondsee (Austria) were analysed directly for the presence of mcyA and microcystin by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry. All filaments assigned to P. rubescens contained mcyA. The proportion of inactive microcystin genotypes in populations with a low (Irrsee) or high density (Mondsee) of P. rubescens was 5% and 21%, each. The results of this study demonstrate that P. rubescens typically contain mcy genes whereas P. agardhii have a patchy distribution of mcy genes. In both species microcystin producers co-occur with non-microcystin producers due to the absence/inactivation of mcy genes.


Know how. Four-layer bandaging.
The concept of four-layer bandaging originated in the 1980s at Charing Cross Hospital, London. Since then, a second four-layer system in kit form has been developed. The results of a recent trial involving 232 patients showed that the two systems have equivalent healing. Today, four-layer bandaging is a popular compression regime, and is used in a wide range of health-care settings. Ease of application of the bandages, and reproducibility, mean that it is not only a therapeutic technique but a very safe one.


How well do prediction equations predict? Using receiver operating characteristic curves and accuracy curves to compare validity and generalizability.
Although morbidity and mortality prediction equations are widely used in planning, clinical practice, and health risk appraisal, their validity and generalizability have been tested only in a limited way. Previous attempts lacked an absolute standard of performance and looked only at the equations' ability to predict who would become ill (sensitivity), not the equally important ability to predict who would remain healthy (specificity). We compared six all-cause mortality prediction equations using receiver operating characteristic curves and accuracy curves, which overcome the limitations of earlier methods and provide a concise visual representation of the results. We used equations from five prospective studies conducted in the United States (Tecumseh at 8 and 12 years of follow-up, Framingham, Chicago Gas, Chicago Western Electric, and Albany), each of which included cholesterol, smoking, and blood pressure as independent variables, to predict 12-year mortality in Tecumseh males age 40-54 years. Previous studies suggested that these equations predict equally well. Our analysis found that, although all predict better than chance, Albany, Chicago Western Electric, and Tecumseh at 8 years underestimate mortality. Receiver operating characteristic and accuracy curves are a promising technique for assessment of prediction equations.


Depot and oral antipsychotics: patient preferences and attitudes are not the same thing.
Some clinicians believe that antipsychotic depot injections are unacceptable to patients. This cross-sectional study investigated patients' attitudes regarding antipsychotics, and included within-participant comparisons. Two hundred and twenty-two out-patients with schizophrenia/schizoaffective disorder completed the Drug Attitude Inventory (DAI-10), scales on insight, side effects and treatment preferences. Formulation preference was associated with current medication formulation: depots were preferred by 43% (33/76) on depot vs 6% (8/146) on orals (P < 0.001). Attitudes (DAI scores) regarding current formulation were influenced by illness duration, extrapyramidal symptoms and insight but not by formulation (depot vs oral). For those with experience of both formulations, participants currently on tablets scored depots less favourably than oral (4.27 vs 6.89, P < 0.001); those on depot did not differentiate. When voluntary patients on maintenance antipsychotics are asked about their attitudes to their current medication, those on depot respond similarly to those on oral. However, when asked to state a preference for formulation (depot vs oral), patients tend to favour their current formulation. Whatever leads some to switch from depot to oral, leaves a lasting negative impression of the depot and this may limit uptake of newer depots.


[Fabrication and evaluation of hydroxyapatite-chitosan scaffold via simulated body fluid biomimetic mineralization].
This research aimed to fabricate a hydroxyapatite (HA)-chitosan scaffold via simulated body fluid (SBF) biomimetic mineralization and determine how mineralization time affects scaffold construction and cell compatibility. The HA-chitosan scaffolds were fabricated by freeze-drying technique and then subjected to precalcification, also known as alternative soaking method. Afterward, precalcificated scaffolds were placed into the SBF to conduct the mineralization process. Mineralization time was set at 7, 14, and 21 days, corresponding to three experimental groups. Pure chitosan scaffolds acted as the control group, and the physical and chemical properties of the four groups were tested. Osteogenic-induced adipose-derived stem cells (ADSCs) were seeded into the scaffolds to investigate the scaffolds' cell compatibility. The mineral substance of the 14-day group exhibited a uniform distribution. The crystal composition of the mineral substance suited the HA's features. The compressive elastic modulus increased along with the extension of mineralization time. The 21-day group showed a statistically significant increase in compressive elastic modulus compared with the control group (P < 0.05). The cell proliferation level of the 14-day group was significantly the highest among the three experimental groups (P < 0.05). The calcium ion and the type I collagen had the highest secretion amount when the cells were seeded into the 14-day group. The SBF biomimetic mineralization method can be used to fabricate HA-chitosan bone-tissue-engineering scaffolds. The biological compatibility, as well as the chemical and physical properties, reached the optimum levels at day 14.


Do children with Blount disease have lower body mass index after lower limb realignment?
Children with Blount disease are typically obese. The goal of our study was to assess whether children with Blount disease had lower body mass index (BMI) after surgical correction of their lower limb deformity. A surgical data base was used to identify children with Blount disease. Demographic information including age of disease onset, ethnicity, health insurance status, and laterality was noted. Preoperative and most recent BMI values were documented. Using full-length standing radiographs, the mechanical axis deviation (MAD) and leg length discrepancy (LLD) were measured preoperatively and at latest follow-up. The relationship of the change in BMI with various demographic and radiologic parameters was evaluated. Fifty-one children (32 males, 19 females) with Blount disease (23 early onset, 28 late onset) affecting 70 lower extremities (32 unilateral and 19 bilateral) underwent a variety of surgical procedures. All 47 children who underwent gradual correction with external fixation also underwent nutritional counseling while receiving inpatient rehabilitation. At an average follow-up of 48 months, MAD improved from 80.5 mm medial to 16.1 mm medial (P<0.0001) and LLD improved from 19.6 to 10.9 mm (P=0.0002). During the same time period, the BMI increased from 35 (95% confidence interval, 32-37) to 38 (95% confidence interval, 35-41; P=0.0006). Compared with their preoperative BMI, 76% of the children had an increase in their BMI at the latest follow-up. There was no association of the change in the patient's BMI with their age of disease onset, sex, ethnicity, health insurance status, final MAD, or LLD. There was a tendency for the patient's BMI to increase with longer follow-up (P=0.002). Using multivariate analysis, only the length of follow-up was associated with an increase in BMI (P=0.026). Despite improvement in limb alignment and LLD after surgery, the BMI of the majority of children with Blount disease increased over time. Other strategies for addressing obesity amongst these children are warranted. Level IV--case series.


Screening of 110 cirrhotic patients for hepatitis B and C at Saidu Teaching Hospital Saidu Sharif Swat.
Cirrhosis liver is amongst the commonest clinical conditions in Pakistan. Besides other causes, it is mostly related to previous viral infection, in this part of the world. The main objective of this study was to find the frequency of HBsAg and anti HCV in cirrhotic patients of Swat district. This Prospective Study was conducted in the Medical unit of Saidu Teaching Hospital, Saidu Sharif from January 2006 to December 2006. One hundred and ten Cirrhotic Patients, 62 Male and 48 Female aged from 14 to 75 years were included in this study. Sera were tested by 3rd generation Elisa method. Fifty-nine out of 110 (53.6%) patients were positive for anti HCV antibodies. Twenty-four (21.81%) Patients were positive for HBsAg. Eleven (10%) patients were positive for both HBsAg and anti HCV anti bodies. Sixteen (14.54%) were negative for both sero maker of hepatitis B and C. It is evident from this study that Hepatitis C, is a leading cause of cirrhosis in District Swat followed by HBV. Both the viruses account for at least three fourths of the total Cirrhotic.


Physical therapy interventions for gross motor skills in people with an intellectual disability aged 6 years and over: a systematic review.
The systematic review was undertaken to investigate the effectiveness of physical therapy interventions for improving gross motor skills (GMSs) in people with an intellectual disability aged 6 years and older. There is a lack of physical therapy research for GMSs in this population, and no prior systematic review. People with an intellectual disability may require specific teaching approaches within therapy interventions to accommodate their cognitive and learning needs. People with an intellectual disability who suffer from GMS deficits can benefit from physical therapy to help improve their GMSs. Data sources were PubMed, CINAHL, Embase, and ProQuest. Reference lists of relevant identified articles were also hand searched. Papers published in English from 1 January 2008 to 22 October 2014 were considered for inclusion. This start date was chosen to reflect the tenets of the United Nations Convention on the Rights of Persons with Disabilities which was ratified in 2008.Eligible study designs for inclusion were randomized controlled trial (RCT), pseudo-RCT, repeated measures, and case report. Overall, 887 potential articles were identified, of which 42 were retrieved for full-text review, and seven were finally included. Critical appraisal was independently conducted by two reviewers using the Joanna Briggs Institute appraisal checklists; no articles were excluded following critical appraisal. Data extraction was performed using Joanna Briggs Institute Meta-Analysis of Statistics Assessment and Review Instrument data extraction instruments. High heterogeneity between the studies precluded meta-analysis of the results; a narrative synthesis was completed instead. Two RCTs, two pseudo-RCTs, two repeated measures studies, and one case report were included. Studies varied in regard to participants' intellectual disability, and the clinical interventions used. Interventions were well tolerated with negligible adverse effects. Significant improvements were reported for cadence and nondimensionalized gait velocity following body-weight-supported gait training; cadence following lower limb strengthening exercises; and the Gross Motor Function Measure-88 measures following adapted judo training. These results suggest that task-specific training may be useful. However, the overall quality of evidence was low. The evidence identified by this systematic review supporting physical therapy for improving GMSs in people with an intellectual disability was limited because of the low quality of studies (only two were RCTs) and only three statistically significant findings. Given the current paucity of evidence, a future systematic review, which sought to identify evidence regarding interventions used by a range of health and education professionals for improving GMSs in people with an intellectual disability, could provide a broader evidence base for clinical interventions which may be utilized by physical therapists.


Dead-time correction of a multi-element SSD for fluorescent XAFS.
The correct evaluation of the live time of a detection system is found to be important for correcting the counting loss of a multi-element detection system for fluorescent XAFS experiments. Synchronous resetting of preamplifiers and the suspension of electronics during the reset period is an effective method. The dead time of the incoming count rate should also be corrected, since its non-linearity cannot be neglected. A 19-element solid-state detector system which can count up to 3.7 x 10(5) counts s(-1) channel(-1) with <270 eV FWHM for Mn Kalpha has been realized; the dead time was independent of the incoming photon energy.


Nitric Oxide-Delivering High-Density Lipoprotein-like Nanoparticles as a Biomimetic Nanotherapy for Vascular Diseases.
Disorders of blood vessels cause a range of severe health problems. As a powerful vasodilator and cellular second messenger, nitric oxide (NO) is known to have beneficial vascular functions. However, NO typically has a short half-life and is not specifically targeted. On the other hand, high-density lipoproteins (HDLs) are targeted natural nanoparticles (NPs) that transport cholesterol in the systemic circulation and whose protective effects in vascular homeostasis overlap with those of NO. Evolving the AuNP-templated HDL-like nanoparticles (HDL NPs), a platform of bioinspired HDL, we set up a targeted biomimetic nanotherapy for vascular disease that combines the functions of NO and HDL. A synthetic S-nitrosylated (SNO) phospholipid (1,2-dipalmitoyl-sn-glycero-3-phosphonitrosothioethanol) was synthesized and assembled with S-containing phospholipids and the principal protein of HDL, apolipoprotein A-I, to construct NO-delivering HDL-like particles (SNO HDL NPs). SNO HDL NPs self-assemble under mild conditions similar to natural processes, avoiding the complex postassembly modification needed for most synthetic NO-release nanoparticles. In vitro data demonstrate that the SNO HDL NPs merge the functional properties of NO and HDL into a targeted nanocarrier. Also, SNO HDL NPs were demonstrated to reduce ischemia/reperfusion injury in vivo in a mouse kidney transplant model and atherosclerotic plaque burden in a mouse model of atherosclerosis. Thus, the synthesis of SNO HDL NPs provides not only a bioinspired nanotherapy for vascular disease but also a foundation to construct diversified multifunctional platforms based on HDL NPs in the future.


Crisaborole and its potential role in treating atopic dermatitis: overview of early clinical studies.
Atopic dermatitis (AD), a chronic, relapsing, inflammatory skin disease that is characterized by intense pruritus and eczematous lesions with up to 90% of patients presenting with mild to moderate disease. Current topical treatments for AD have not changed in over 15 years and are associated with safety concerns. In AD, overactivity of phosphodiesterase 4 (PDE4), leads to inflammation and disease exacerbation. Crisaborole Topical Ointment, 2%, is a novel, nonsteroidal, topical anti-inflammatory PDE4 inhibitor currently being investigated for the treatment of mild to moderate AD. Preliminary studies in children and adults demonstrated favorable efficacy and safety profiles. Crisaborole may represent an anti-inflammatory option that safely minimizes the symptoms and severity of AD and that can be used for both acute and long-term management.


Primary dermatofibrosarcoma protuberans of orbit--a rare entity.
Primary Dermatofibrosarcoma Protuberance (DFSP) is a rare neoplasm of dermal origin. Though it is a locally aggressive tumor with high recurrence rate, however distant metastasis can also occur. Orbital DFSP is an uncommon phenomenon. It has been reported due to distant metastasis or invasion from adjacent structures but Primary Orbital DFSP is a unique entity in itself. Herein we report a rare case of primary DFSP of the orbit in a 70-year- old lady who underwent orbital exenteration. Histopathology examination (HPE) revealed spindle cells arranged in storiform pattern and immunohistochemistry (IHC) revealed CD34 positive and S100 negative.


Inhaled corticosteroids in chronic obstructive pulmonary disease.
The effectiveness of inhaled corticosteroids (ICS) in patients with chronic obstructive pulmonary disease (COPD) remains controversial. Randomized controlled trials, meta-analyses, medication withdrawal studies, and observational reports have examined this question, with mixed results. Observational studies have been subject to criticism because of study design involving immortal time bias. Some randomized controlled trials suggest small benefits in lung function and health status, and a reduction in the rate of acute exacerbations of COPD and mortality, but their incomplete follow-up and statistical methods have been criticized. The greatest benefits of ICS in COPD have been reported with use of ICS and long-acting beta-agonist combination therapy, although no benefit was found for the primary outcome studied under the most rigorous methodology by the recent TORCH and Optimal randomized trials. Thus, although future randomized trials will need to be conducted with the most rigorous methodology for all outcomes, much uncertainty remains regarding the potential benefits of ICS in COPD.


A Specific Strain of Lactic Acid Bacteria, Lactobacillus paracasei, Inhibits Inflammasome Activation In Vitro and Prevents Inflammation-Related Disorders.
Some strains of lactic acid bacteria (LAB) have anti-inflammatory effects, but the mechanism underlying the alleviation of inflammation by LAB is not fully understood. In this study, we examined the inhibitory effect of a certain strain of LAB, Lactobacillus paracasei, on inflammasome activation, which is associated with various inflammatory disorders. Using bone marrow-derived macrophages from BALB/c mice, we found that L. paracasei, but not L. rhamnosus, suppressed NLRP3 inflammasome activation and inhibited subsequent caspase-1 activation and IL-1β secretion. L. paracasei also had inhibitory effects on AIM2 and NLRC4 inflammasome activation as well as the NLRP3 inflammasome. These inhibitory effects of L. paracasei on inflammasome activation were dependent on autocrine IL-10 induced by L. paracasei-stimulated macrophages. Furthermore, IL-10 production by L. paracasei-stimulated macrophages was involved with phagocytosis and the NOD2 signaling pathway in macrophages. In addition to in vitro studies, oral administration of L. paracasei in C57BL/6 mice reduced monosodium urate crystal-induced peritoneal inflammation in vivo. Moreover, continuous intake of L. paracasei in C57BL/6 mice alleviated high fat diet-induced insulin resistance and aging-induced expression of biomarkers for T cell senescence. Taken together, we demonstrated that L. paracasei inhibits inflammasome activation in vitro and exhibits an anti-inflammatory function in vivo. These results indicate that LAB that have inhibitory effects on inflammasome activation might contribute to the alleviation of inflammation-related disorders.


Differential activation of interferon-inducible genes by hepatitis C virus core protein mediated by the interferon stimulated response element.
We previously found that hepatitis C virus (HCV) core protein, which possesses the consensus sequence of genotype 1b, transcriptionally activates the interferon (IFN)-inducible 2'-5'-oligoadenylate synthetase (2'-5'-OAS) gene in human hepatocyte cells. To clarify the mechanism of this activation, we further characterized the core protein as an activator of the 2'-5'-OAS gene. We demonstrated that the activation of the 2'-5'-OAS gene by the core protein is a general phenomenon, regardless of HCV genotype and strain. We showed that the 20 N-terminal amino acids (aa) of the core protein were important to the activation of the 2'-5'-OAS gene, although this N-terminal region did not have any effect on the subcellular localization of the core protein. We demonstrated that the core protein was able to activate all promoters possessing the IFN-stimulated response element (ISRE) examined. However, we found that the level of activation of the 2'-5'-OAS gene promoter possessing a particular variant type of ISRE was significantly higher than that of other IFN-inducible gene promoters. This phenomenon was confirmed using synthetic promoters possessing five repeats of the consensus or a 2'-5'-OAS-type ISRE. In addition, we showed that gene activation induced by the core protein is mediated by the ISRE. These results imply that the core protein prefers a subclass of IFN-inducible genes, the promoters of which possess the 2'-5'-OAS-type ISRE. Accordingly, we found that the IFN-inducible double-stranded RNA-specific adenosine deaminase gene promoter, possessing a 2'-5'-OAS-type ISRE sequence, was also efficiently activated by the core protein. The exact mechanism by which the core protein enhances gene expression was not determined, but we could find no effects of core protein on gene expression and phosphorylation status of the components of the JAK-STAT signaling transduction pathway.


Vegetarianism and other eating practices among youth and young adults in major Canadian cities.
To estimate the prevalence and sociodemographic characteristics of youth and young adults in major Canadian cities with self-reported vegetarian dietary practices and examine efforts to alter their diets. Data were collected in autumn 2016 via web-based surveys. Respondents reported vegetarian dietary practices (vegan, vegetarian or pescatarian) and efforts in the preceding year to consume more or less of several nutrients, food groups and/or foods with particular attributes. Logistic regression models examined sociodemographic correlates of each vegetarian dietary practice and differences in other eating practices by diet type. Participants were recruited from five major Canadian cities. Youth and young adults, aged 16-30 years (n 2566). Overall, 13·6 % of respondents reported vegetarian dietary practices: 6·6 % vegetarian, 4·5 % pescatarian and 2·5 % vegan. Sex, race/ethnicity, self-reported frequency of using the Nutrition Facts table and health literacy were significantly correlated with self-reported vegetarian dietary practice (P < 0·01 for all). Efforts to consume more fruits and vegetables (66·8 %) and protein (54·8 %), and less sugar (61·3 %) and processed foods (54·7 %), were prevalent overall. Respondents with vegetarian dietary practices were more likely to report efforts to consume fewer carbohydrates and animal products, and more organic, locally produced, ethically sourced/sustainably sourced/fair trade and non-GM foods (P < 0·01 for all), compared with those without these reported dietary practices. Nearly 14 % of the sampled youth and young adults in major Canadian cities reported vegetarian dietary practices and may be especially likely to value and engage in behaviours related to health-conscious diets and sustainable food production.


Challenges in Pathology Specimen Processing in the New Era of Precision Medicine.
Precision therapies for patients with driver mutations can offer deep and durable responses that correlate with diagnosis, metastasis prognosis, and improvement in survival. The use of such targeted therapies will continue to increase, pushing us to change our traditional approaches. We needed to search for new tools to effectively integrate technological advancements into our practices because of their capability to improve the efficiency and accuracy of our diagnostic and treatment approaches. Perhaps nothing is as relevant as identifying and implementing new workflows for processing pathologic specimens and for improving communication of critical laboratory information to and from clinicians for appropriate care of patients in an efficient and timely manner. To define the gold standard in delivering the best care for patients, to identify gaps in the process, and to identify potential solutions that would improve our process, including gaps related to knowledge, skills, attitudes, and practices. We assembled a multidisciplinary team to systematically perform a gap analysis study to clarify the discrepancy between the current reality in pathology specimen processing and the desired optimal situation to deliver the results intended for patient care. A practical collaborative workflow for specimen management that seeks the cooperation of stakeholders in each medical discipline to provide guidelines in specimen collection, delivery, processing, and reporting of results with the ultimate goal of improving patient outcomes is provided. New tools are required to effectively integrate data-driven approaches in specimen processing to meet the new demands.


RNA content and volume of motor neurons in amyotrophic lateral sclerosis. II. The lumbar intumescence and nucleus dorsalis.
The content of RNA and volume of individual neurons isolated from the nucleus dorsalis and from the ventrolateral portion of the lumbar swelling were determined in eight cases of amyotrophic lateral sclerosis (ALS) and eight controls whose spinal cords were obtained at autopsy. The mean content of RNA in the lumbar motor neurons of the controls was 557 pg, compared to only 386 pg in the ALS group. This represents approximately a 31% reduction and is highly significant, p less than 0.01. No difference in RNA content was observed between the ALS group and controls in the nucleus dorsalis, which suggests that the reduction of RNA is restricted to the motor system in ALS. The volume of individual motor neurons of the lumbar intumescence was not significantly different between the controls and ALS.


Expression, Purification, and Antimicrobial Activity of S100A12.
Calgranulin proteins are important mediators of innate immunity and are members of the S100 class of the EF-hand family of calcium binding proteins. Some S100 proteins have the capacity to bind transition metals with high affinity and effectively sequester them away from invading microbial pathogens in a process that is termed "nutritional immunity". S100A12 (EN-RAGE) binds both zinc and copper and is highly abundant in innate immune cells such as macrophages and neutrophils. We report a refined method for the expression, enrichment and purification of S100A12 in its active, metal-binding configuration. Utilization of this protein in bacterial growth and viability analyses reveals that S100A12 has antimicrobial activity against the bacterial pathogen, Helicobacter pylori. The antimicrobial activity is predicated on the zinc-binding activity of S100A12, which chelates nutrient zinc, thereby starving H. pylori which requires zinc for growth and proliferation.


Cardiovascular effects of a new positive inotropic agent, (-)-(R)-1-(p-hydroxyphenyl)-2-[(3,4-dimethoxyphenethyl)amino]-ethanol (TA-064) in the anesthetized dog and isolated guinea pig heart.
The positive inotropic effect of TA-064, (-)-(R)-1-(p-hydroxyphenyl)-2-[(3,4-dimethoxyphenethyl)amino]ethanol, was studied in the anesthetized dog and isolated guinea pig heart. An intravenous administration of TA-064 dose-dependently increased the cardiac contractile force with little effect on blood pressure in dogs. The positive inotropic activity of TA-064 was 1/100 that of isoproterenol and similar to that of dobutamine. This effect of TA-064 was stereospecific, and it was blocked by practolol. Thus TA-064 has beta 1-adrenoceptor agonistic action. The positive inotropic effect of TA-064 was more pronounced than the positive chronotropic effect, compared with those of isoproterenol. Similar effect of TA-064 was observed in the reserpinized dog and in the isolated perfused heart of the guinea pig as well. TA-064 administered intraduodenally at a dose of 0.1 mg/kg increased contractile force by 120% of the control, and the effect lasted for more than 150 min. TA-064 given in the femoral artery demonstrated a very weak vasodilating effect on the artery. TA-064 is an orally active, positive inotropic agent. The selective positive inotropic action of TA-064 may result from its beta 1-adrenoceptor agonistic property.


CD30-Positive T-Cell Lymphoproliferative Disease of the Oral Mucosa in Children: A Manifestation of Epstein-Barr Virus-Associated T-Lymphoproliferative Disorder.
Eosinophilic ulcer of the oral mucosa (EUOM) is a very rare, benign, self-limiting ulcerative lesion of the oral cavity of unknown pathogenesis, and belongs to the same spectrum of CD30(+) T-cell lymphoproliferative disease (LPD) of the oral mucosa. The etiology and pathogenesis of the disease are unknown. We report two cases in children who were initially diagnosed with EUOM and CD30(+) T-cell LPD, respectively. However, retrospective analysis revealed that a majority of infiltrated atypical T cells were positive for Epstein-Barr virus (EBV). The present cases suggest that the pathogenesis and etiology of EUOM or CD30(+) T-cell LPD occurring in children are different from those in adults. EUOM or CD30(+) T-cell LPD in children is a manifestation of EBV-positive T-cell LPD, and should therefore be distinguished from the disease in adults.


Gastric Invasive Micropapillary Carcinoma with Intestinal Phenotypes Harboring a TP53 R175H Mutation.
We report a case of gastric invasive micropapillary carcinoma (IMPC) in an 86-year-old female patient. She was admitted to our hospital with a chief complaint of bloody emesis. Upper gastrointestinal endoscopy found a gastric adenocarcinoma at the antrum. The biopsy specimens showed moderately differentiated adenocarcinoma with invasive small tumor nests. Distal gastrectomy with systematic lymph node dissection demonstrated that the tumor had IMPC through a pathological examination. Despite the depth of tumor invasion (the submucosa), extensive lymph node metastases were observed. Anti-D2-40 immunostaining revealed numerous infiltrating tumor cell nests in the lymphatic vessels, which could explain subsequent multiple lymph node metastases. The adenocarcinoma showed intestinal phenotypes by several immunohistochemical studies. One of these antibodies (CD10) clearly demonstrated the inverted apical-basal (inside-out) pattern of IMPC, whereas it showed an ordinary pattern in intestinal metaplasia adjacent to the tumor. Furthermore, genetic analysis by direct sequencing revealed a point mutation in the exon 5 of TP53 in the tumor. The mutation presumably harbors a missense mutation from Arg to His at the codon 175 (R175H). R175H has been previously described as a 'gain-of-function' mutation with a high invasive or metastatic potential in several types of cancers. In summary, this is one of the first reported cases of gastric IMPC with intestinal phenotypes harboring a TP53 R175H mutation in the literature.


Protein kinase A mediates inhibition of the thrombin-induced platelet shape change by nitric oxide.
The thrombin-induced platelet shape change was blocked by nitric oxide (NO), as revealed by scanning electron microscopy, light transmission, and resistive-particle volume determination. The inhibitory effect of NO was accompanied by an increase in levels of both cyclic guanosine monophosphate (cGMP) and cyclic adenosine monophosphate (cAMP) and phosphorylation of the vasodilator-stimulated phosphoprotein (VASP). However, the inhibition of the shape change was only mimicked by cAMP analogs (Sp-5,6-DClcBIMPS, 8-AHA-cAMP, and 8-CPT-cAMP) and not by cGMP analogs (8-Br-PET-cGMP, 8-Br-cGMP, and 8-pCPT-cGMP). The effect of NO on the thrombin-induced shape change was prevented by the protein kinase A (PKA) antagonists Rp-8-Br-cAMPS and Rp-cAMPS. The protein kinase G (PKG) antagonist Rp-8-CPT-cGMPS strongly inhibited PKG-mediated 46-kDa VASP Ser239 phosphorylation, but did not inhibit the thrombin-induced shape change or the PKA-mediated VASP Ser157 phosphorylation. Whereas an inhibitor of cyclic nucleotide phosphodiesterase (PDE) 3A (milrinone) mimicked the effect of NO, inhibitors of PDE2 (erythro-9-(2-hydroxy-3-nonyl)adenine) and PDE5 (dipyridamole) were poorly effective. We concluded that (1) NO was a potent and reversible inhibitor of the platelet shape change, (2) the shape change was reversible, (3) the inhibitory effect of NO was mediated through activation of PKA, (4) the onset of the NO effect coincided with VASP Ser157 phosphorylation, and (5) removal of NO and platelet shape change coincided with VASP Ser157 dephosphorylation. These findings are compatible with elevation of cGMP by NO in a compartment close to PDE3A, PKA, and VASP, leading to a local increase of cAMP able to block thrombin-induced shape change.


Triplet excited States as a source of relevant (bio)chemical information.
The properties of triplet excited states are markedly medium-dependent, which turns this species into valuable tools for investigating the microenvironments existing in protein binding pockets. Monitoring of the triplet excited state behavior of drugs within transport proteins (serum albumins and α1-acid glycoproteins) by laser flash photolysis constitutes a valuable source of information on the strength of interaction, conformational freedom and protection from oxygen or other external quenchers. With proteins, formation of spatially confined triplet excited states is favored over competitive processes affording ionic species. Remarkably, under aerobic atmosphere, the triplet decay of drug@protein complexes is dramatically longer than in bulk solution. This offers a convenient dynamic range for assignment of different triplet populations or for stereochemical discrimination. In this review, selected examples of the application of the laser flash photolysis technique are described, including drug distribution between the bulk solution and the protein cavities, or between two types of proteins, detection of drug-drug interactions inside proteins, and enzyme-like activity processes mediated by proteins. Finally, protein encapsulation can also modify the photoreactivity of the guest. This is illustrated by presenting an example of retarded photooxidation.


Regioselective Diversification of 2,1-Borazaronaphthalenes: Unlocking Isosteric Space via C-H Activation.
Methods for the regioselective C-H borylation and subsequent cross-coupling of the 2,1-borazaronaphthalene core are reported. Azaborines are dependent on B-N/C═C isosterism when employed in strategies for developing diverse heterocyclic scaffolds. Although 2,1-borazaronaphthalene is closely related to naphthalene in terms of structure, the argument is made that the former has electronic similarities to indole. Based on that premise, iridium-mediated C-H activation has enabled facile installation of a versatile, nucleophilic coupling handle at a previously inaccessible site of 2,1-borazaronaphthalenes. A variety of substituted 2,1-borazaronaphthalene cores can be successfully borylated and further cross-coupled in a facile manner to yield diverse C(8)-substituted 2,1-borazaronaphthalenes.


Indolinooxazolidine: a versatile switchable unit.
The design of multiresponsive systems continues to arouse a lot of interest. In such multistate/multifunctional systems, it is possible to isomerize a molecular system from one metastable state to another by application of different stimulation such as light, heat, proton, or electron. In this context, some researches deal with the design of multimode switch where a same interconversion between two states could be induced by using indifferently two or more different kind of stimuli. Herein, we demonstrate that the association of an indolinooxazolidine moiety with a bithiophene unit allows the development of a new trimode switch. A reversible conversion between a colorless closed form and a colorful open form can be equally performed by light, proton, or electrical stimulation. In addition, the oxidation of this system allows the generation of a third metastable state.


Comparison of empagliflozin and sitagliptin therapy on myocardial perfusion reserve in diabetic patients with coronary artery disease.
Sodium-glucose co-transporter 2 inhibitors reduce the risk of cardiovascular events in type 2 diabetic patients with coronary artery disease (CAD); however, the underlying mechanisms remain unclear. We compared the effects of empagliflozin vs. sitagliptin therapy on myocardial perfusion reserve (MPR) using dynamic single-photon emission computed tomography (SPECT) imaging. In total, 100 patients with type 2 diabetes, CAD and an MPR <2.5 were randomized to receive either empagliflozin (10 mg once daily) or sitagliptin (100 mg once daily). Dynamic SPECT examinations were performed at baseline and at 6 months. The primary endpoint was the percent change of global MPR. Evaluable SPECT data were available for 98 patients. Baseline clinical characteristics and SPECT data were well balanced between the two groups. At a 6-month follow-up, the fasting glucose and glycated hemoglobin levels significantly decreased in both groups. Hematocrit and hemoglobin levels significantly increased in the empagliflozin group but not in the sitagliptin group. The global MPR significantly improved after treatment in both groups (34.5 ± 70.6%; P = 0.005 for empagliflozin vs. 22.4 ± 45.7%; P = 0.024 for sitagliptin). However, there was no significant difference in the global MPR between the two groups (P = 0.934). Similar findings were detected with regard to the regional MPR. Among patients with type 2 diabetes and CAD, both empagliflozin and sitagliptin significantly improved the global MPR with no significant difference between the groups.


Caralluma fimbriata and metformin protection of rat pancreas from high fat diet induced oxidative stress.
A high fat diet promotes oxidative stress, which contributes to the development of pancreatic fibrosis. We compared the protective effects of a hydroalcoholic extract of Caralluma fimbriata (CFE) to metformin (Met) in the pancreas of Wistar rats fed a high fat diet. The experimental animals were divided into five groups: control (C), treated with CFE (C + CFE), treated with high fat diet (HFD), high fat diet treated with CFE (HFD + CFE), and high fat diet treated with metformin (Met) (HFD + Met). CFE was administered orally to groups C + CFE and HFD + CFE rats for 90 days. Met was given to the HFD + Met group. After 90 days, oxidative stress markers in the pancreas including reduced glutathione (GSH), lipid oxidation (LO), protein oxidation (PO), and activities of antioxidant and polyol pathway enzymes, aldose reductase (AR) and sorbitol dehydrogenase (SDH) were assayed and tissue histology was examined. Establishment of oxidative stress in high fat diet fed rats was verified by elevated LO and PO, decreased GSH, decreased activities of antioxidants and increased activities of polyol pathway enzymes. Oxidative stress was prevented in HFD + CFE and HFD + Met groups. Group C + CFE exhibited improved antioxidant status compared to group C. CFE treatment prevented high fat diet induced acinar cell degeneration, necrosis, edema and hemorrhage. CFE could be used as adjuvant therapy for preventing or managing high fat diet induced pancreatic damage.


Prevention by a stable prostacyclin analogue, 7-oxoprostaglandin I2 of the platelet loss during experimental cardiopulmonary bypass.
A great percentage of circulating platelets is lost through activation and aggregation during total body perfusion used in cardiac surgery. The effect on platelet count of a chemically stable prostacyclin analogue, 7-oxoprostaglandin I2 was investigated in anaesthetized dogs subjected to cardiopulmonary bypass of 120 min duration. In control (untreated) dogs the perfusion decreased the amount of circulating platelets by more than 60% of their initial value. This loss was completely prevented by i.v. infusion of the analogue (50 micrograms.kg-1.min-1) without affecting the haemodynamic state of the preparation. With the infusion of native prostacyclin at the same rate, a similar protective action was found to be associated with significant arterial hypotension. It was concluded that 7-oxoprostaglandin I2 is an useful pharmacological tool for maintaining physiological platelet counts and thus preserving haemostatic stability in open heart surgery.


Identifying and locating pharmacist certificate programs, traineeships, and certification agencies.
To provide a list of available certification and certificate programs that can be completed to enhance or validate pharmacists' knowledge and skills in the treatment of specific diseases. Medline/PubMed searched using the terms "pharmacist" and "certification" or "pharmacist" and "credentialing"; Web sites of national pharmacy organizations. No information was found in Medline/PubMed about the value of certification or certificate programs for pharmacists. A comprehensive, but not exclusive, list of pharmacist certificate programs, traineeships, and certifications available is provided, as well as guidance on where to find information about such programs. Pharmacists have many opportunities for training in specialized areas that can broaden their knowledge and skills to improve patient care, provide credentialing for reimbursement, and advance their professional status.


Immunotherapy in allergy to dog: a double-blind clinical study.
Twenty-seven asthmatic children allergic to dog were included in the study. Their allergy to dog was confirmed by positive results from the skin prick, radioallergosorbent and provocation tests. The subjects were randomly assigned to active and placebo groups. Fifteen subjects received immunotherapy with a commercial standardized aluminum hydroxide-bound dog dander extract and 12 subjects received placebo injections containing histamine. After one year's therapy the conjunctival sensitivity to dog dander extract had decreased significantly (p less than 0.001) in the active treatment group compared to the placebo group. The hyposensitization effect was already observable after six to eight months of treatment. The decrease in bronchial sensitivity was less marked than that in conjunctival sensitivity and statistically not significant. Immunotherapy was observed to be safe and effective. No systemic adverse reactions occurred and local reactions were mild.


Endocrine abnormalities in human temporal lobe epilepsy.
Patients with temporal lobe epilepsy secrete ACTH at higher rates and in greater amounts than normal subjects. Temporal lobectomy restores ACTH secretion to normal amounts and rates. The ACTH secretion in temporal lobe epilepsy is independent of anticonvulsant drug effect and seizure frequency. Electrical stimulation of medial temporal lobe structures in patients with temporal lobe epilepsy affected ACTH secretion in a manner consistent with the hypothesis that ACTH secretion is regulated by tonic inhibition. A defect in the excitatory and/or inhibitory components of this regulatory process appears to exist in temporal lobe epilepsy.


Age dependence of the gamma-carboxyglutamic acid containing proteins.
A distinct age dependence was observed in the occurrence of the gamma-carboxyglutamic acid containing proteins obtained from kidney cortex, liver and femoral bone. In the femoral bone, a stepped increase was seen when comparing animals 60, 350 and 700 days old. In the liver and mainly in the kidney cortex there was no change during the first year of life, but a distinct increase in the gamma-carboxyglutamate containing proteins was seen between day 350 and 700. It was also established that the increased level of gamma-carboxyglutamate at least in kidney cortex is due to a more intense carboxylation reaction in older animals as increased incorporation of labelled bicarbonate was observed in the microsomal fraction. In accordance with our previous results the occurrence of the carboxyglutamic acid containing proteins was shown to be vitamin D dependent; in vitamin D deprived animals supplemented with this vitamin the reappearance of gamma-carboxylate is more distinct in situations where higher levels of gamma-carboxyglutamic acid occur physiologically.


Gestational diabetes diagnostic methods (GD2M) pilot randomized trial.
To test the feasibility of conducting a pragmatic randomized controlled trial (RCT) comparing the International Association of Diabetes in Pregnancy Study Groups (IADPSG) versus Carpenter-Coustan diagnostic criteria for gestational diabetes (GDM), and to examine patient and provider views on GDM screening. A single-blinded pragmatic pilot RCT. Participants with a singleton pregnancy between 24 and 28 weeks gestation received a 50 g oral glucose challenge test and if the value was <200 mg/dL were randomized to either the 2 h 75 g OGTT using the IADPSG criteria or the 3 h 100 g OGTT using the Carpenter-Coustan criteria. Primary outcome was the feasibility of randomization and screening. Secondary outcomes included patient and provider views (or preferences) on GDM testing. Sixty-eight women were recruited, 48 (71 %) enrolled and 47 (69 %) were randomized. Participants in both study arms identified the main challenges to GDM testing to be: drinking the glucola, fasting prior to testing, waiting to have blood drawn, and multiple venipuntures. Women in both study arms would prefer the 2 h 75 g OGTT or whichever test is recommended by their doctor in a future pregnancy. Physicians and nurse midwives endorsed screening and were comfortable with being blinded to the GDM testing strategy and results values. Both pregnant women and providers value GDM screening, and pregnant women can be recruited to a blinded, randomized GDM screening trial with minimal attrition and missing data.


Destabilization of the dystrophin-glycoprotein complex without functional deficits in alpha-dystrobrevin null muscle.
Alpha-dystrobrevin is a component of the dystrophin-glycoprotein complex (DGC) and is thought to have both structural and signaling roles in skeletal muscle. Mice deficient for alpha-dystrobrevin (adbn(-/-)) exhibit extensive myofiber degeneration and neuromuscular junction abnormalities. However, the biochemical stability of the DGC and the functional performance of adbn(-/-) muscle have not been characterized. Here we show that the biochemical association between dystrophin and beta-dystroglycan is compromised in adbn(-/-) skeletal muscle, suggesting that alpha-dystrobrevin plays a structural role in stabilizing the DGC. However, despite muscle cell death and DGC destabilization, costamere organization and physiological performance is normal in adbn(-/-) skeletal muscle. Our results demonstrate that myofiber degeneration alone does not cause functional deficits and suggests that more complex pathological factors contribute to the development of muscle weakness in muscular dystrophy.


Symptom clusters: the new frontier in symptom management research.
The majority of clinical studies on pain, fatigue, and depression associated with cancer are focused on one symptom. Although this approach has led to some advances in our understanding of a particular symptom, patients rarely present with a single symptom. Therefore, even though research focused on single symptoms needs to continue, it is imperative that symptom management research begins to focus on evaluating multiple symptoms, using cross-sectional and longitudinal study designs. In addition, research needs to focus on evaluating the relationships among multiple symptoms, specific interventions, and patient outcomes. One of the initial challenges in research regarding multiple symptoms is the terminology that should be used to describe the concept (e.g., symptom cluster, symptom constellation). Another significant area related to this aspect of symptom management research is determining the nature of clinically significant clusters of symptoms and their associated prevalence rates. Equally important is the need to determine what types of tools/instruments will provide the most valid and reliable data for the assessment of symptom clusters. Other areas that need to be considered as related to the assessment of symptom clusters include the establishment of cut points for symptom severity that would qualify a symptom for inclusion in a cluster; the focus of the assessment; and the choice of the outcome measures that will be used to judge the effect of a symptom cluster on the patient. In the area of intervention studies for symptom clusters, research will need to build on the limited number of clinical trials with single symptoms. Additional considerations related to research on symptom clusters include the determination of the mechanisms underlying the development of symptom clusters; the timing of the measurements for symptom clusters; and statistical challenges in the evaluation of symptom clusters. Research on symptom clusters in patients with cancer is cutting-edge science and a new frontier in symptom management research, and it needs to be done in tandem with research on single symptoms.


N-terminal pro-brain natriuretic peptide and associated factors in the general working population: a baseline survey of the Uranosaki cohort study.
Few data on clinical characteristics associated with N-terminal pro-brain natriuretic peptide (NT-proBNP) or the clinical value of measuring NT-proBNP in the working population are available. The aim of the present study was to investigate the levels of NT-proBNP and their association with clinical variables in the Japanese general working population by using baseline data from the Uranosaki cohort study. In the study, the plasma concentration of NT-proBNP and some biomarkers were measured in addition to the standard health checkups at the workplace. Questionnaires regarding health-related quality of life (HR-QOL) were also completed. A total of 2140 participants were enrolled in the study. Plasma levels of NT-proBNP were positively associated with age, female sex, systolic blood pressure, pulse pressure, prevalent hypertension, smoking habit, high-density lipoprotein cholesterol (HDL-C), and prevalent proteinuria, and negatively associated with body mass index, lipid profiles except HDL-C, uric acid, renal function, and hemoglobin. Both the plasma concentration of high-molecular weight adiponectin and that of high-sensitivity troponin T were positively and independently associated with NT-proBNP. In addition, the HR-QOL score regarding sleep disorder was independently associated with NT-proBNP. Thus, we have obtained evidence that the plasma NT-proBNP is affected by several clinical variables in the general working population.


Assessing food safety and associated food handling practices in street food vending.
Street vendors in the city of Bloemfontein were investigated in order to assess the microbiological quality of the food being sold as well as the level of hygiene conditions under which these food stalls operate. The food samples which were collected included beef, chicken and gravy, while surface samples were taken from the food preparation tables and the hands of the vendors. A structured questionnaire and checklist were used in interviews to determine the status of the vending sites and associated food handling practices. The overall microbiological quality of the foods served by the street vendors was found to be within acceptable safety limits, although the presence of specific microorganisms such as Escherichia coli, Staphylococcus aureus, Salmonella and yeasts is indicative of a degree of ignorance on the part of the food handlers towards proper hygienic practices.


Variations in DNA aneuploid cell content during tumor dissociation in human colon and head and neck cancers analyzed by flow cytometry.
Experimental research involving human solid tumors often requires single cell suspensions of high yield that are representative of the tissue of origin and in which the cellular property of interest is preserved. This is particularly necessary for the determination of DNA ploidy by flow cytometry. Mechanical dissaggregation and proteolytic enzyme digestion are the most commonly employed dissociation techniques for solid tumors. Comparative testing of techniques is often not performed. Mechanical and proteolytic enzyme dissociation techniques were comparatively tested in 77 human squamous cell cancers of the head and neck (SCCHN) and 25 human colon cancers for cellular yield, dye exclusion viability, quality, and morphology of DNA histograms, and the presence and proportion of DNA aneuploid subpopulations. Significant and consistent DNA aneuploid subpopulation losses were noted in mechanical preparations of SCCHN and enzymatic preparations of colon cancers. The frequency of SCCHN specimens with DNA aneuploid subpopulations was underestimated by 52% in mechanical cell suspensions, and the proportion of DNA aneuploid cells was diminished in an additional 30% of the specimens. Conversely, the frequency of specimens with DNA aneuploid subpopulations was underestimated by 38% in cell suspensions from enzymatically dissociated human colon cancer and their proportion diminished in an additional 50% of the specimens. Incubations of human colon cancers with three commonly employed proteolytic enzymes demonstrated a progressive loss of DNA aneuploid subpopulations as a function of enzyme concentration and incubation time. This is a serious potential source of error in the flow cytometric determination of DNA ploidy in human solid tumors, and may contribute to the diversity of results obtained and occasional contradictory conclusions reached in such studies.


Dissemination in cutaneous leishmaniasis. II. Satellite papules and subcutaneous induration.
In a field study of cutaneous leishmaniasis (CL) due to Leishmania major, zymodeme Lon-4, in an endemic focus in Saudi Arabia, 80 patients in a group of 643 patients (12.44%) were found to have multiple, inflammatory, satellite papules (SP) around one or more CL lesions. The SP often appeared to erupt after commencement of antileishmanial treatment. They showed a range of morphology and pathology, and comparisons with corresponding CL lesions showed important differences. Amastigotes were seen in only one of seven biopsy specimens. Another morphologic feature, subcutaneous induration (SCI), was noted on routine palpation in 20 patients in the same patient group (3.11%). The SCI either was radiating all around the lesion and appeared as an "iceberg nodule" or was present as a "tonguelike" process proximal to the CL lesion. Amastigotes were seen in four of nine biopsy specimens of SCI. SP and SCI represent reactions to local dissemination of the parasite or its antigenic products, and as morphologic features aid in the clinical diagnosis of CL.


Electrospun poly(L-lactide) fiber with ginsenoside rg3 for inhibiting scar hyperplasia of skin.
Hypertrophic scarring (HS) has been considered as a great concern for patients and a challenging problem for clinicians as it can be cosmetically disfiguring and functionally debilitating. In this study, Ginsenoside Rg3/Poly(l-lactide) (G-Rg3/PLLA) electrospun fibrous scaffolds covering on the full-thickness skin excisions location was designed to suppress the hypertrophic scar formation in vivo. SEM and XRD results indicated that the crystal G-Rg3 carried in PLLA electrospun fibers was in amorphous state, which facilitates the solubility of G-Rg3 in the PLLA electrospun fibrous scaffolds, and solubility of G-Rg3 in PBS is increased from 3.2 µg/ml for pure G-Rg3 powders to 19.4 µg/ml for incorporated in PLLA-10% fibers. The released G-Rg3 content in the physiological medium could be further altered from 324 to 3445 µg in a 40-day release period by adjusting the G-Rg3 incorporation amount in PLLA electrospun fibers. In vitro results demonstrated that electrospun G-Rg3/PLLA fibrous scaffold could significantly inhibit fibroblast cell growth and proliferation. In vivo results confirmed that the G-Rg3/PLLA electrospun fibrous scaffold showed significant improvements in terms of dermis layer thickness, fibroblast proliferation, collagen fibers and microvessels, revealing that the incorporation of the G-Rg3 in the fibers prevented the HS formation. The above results demonstrate the potential use of G-Rg3/PLLA electrospun fibrous scaffolds to rapidly minimize fibroblast growth and restore the structural and functional properties of wounded skin for patients with deep trauma, severe burn injury, and surgical incision.


Pulsed diode laser-based monitor for singlet molecular oxygen.
Photodynamic therapy (PDT) is a promising cancer treatment. PDT uses the affinity of photosensitizers to be selectively retained in malignant tumors. When tumors, pretreated with the photosensitizer, are irradiated with visible light, a photochemical reaction occurs and tumor cells are destroyed. Oxygen molecules in the metastable singlet delta state O2(1Delta) are believed to be the species that destroys cancerous cells during PDT. Monitoring singlet oxygen produced by PDT may lead to more precise and effective PDT treatments. Our approach uses a pulsed diode laser-based monitor with optical fibers and a fast data acquisition system to monitor singlet oxygen during PDT. We present results of in vitro singlet oxygen detection in solutions and in a rat prostate cancer cell line as well as PDT mechanism modeling.


Ecological Stability Properties of Microbial Communities Assessed by Flow Cytometry.
Natural microbial communities affect human life in countless ways, ranging from global biogeochemical cycles to the treatment of wastewater and health via the human microbiome. In order to probe, monitor, and eventually control these communities, fast detection and evaluation methods are required. In order to facilitate rapid community analysis and monitor a community's dynamic behavior with high resolution, we here apply community flow cytometry, which provides single-cell-based high-dimensional data characterizing communities with high acuity over time. To interpret time series data, we draw inspiration from macroecology, in which a rich set of concepts has been developed for describing population dynamics. We focus on the stability paradigm as a promising candidate to interpret such data in an intuitive and actionable way and present a rapid workflow to monitor stability properties of complex microbial ecosystems. Based on single-cell data, we compute the stability properties resistance, resilience, displacement speed, and elasticity. For resilience, we also introduce a method which can be implemented for continuous online community monitoring. The proposed workflow was tested in a long-term continuous reactor experiment employing both an artificial and a complex microbial community, which were exposed to identical short-term disturbances. The computed stability properties uncovered the superior stability of the complex community and demonstrated the global applicability of the protocol to any microbiome. The workflow is able to support high temporal sample densities below bacterial generation times. This may provide new opportunities to unravel unknown ecological paradigms of natural microbial communities, with applications to environmental, biotechnological, and health-related microbiomes. IMPORTANCE Microbial communities drive many processes which affect human well-being directly, as in the human microbiome, or indirectly, as in natural environments or in biotechnological applications. Due to their complexity, their dynamics over time is difficult to monitor, and current sequence-based approaches are limited with respect to the temporal resolution. However, in order to eventually control microbial community dynamics, monitoring schemes of high temporal resolution are required. Flow cytometry provides single-cell-based data in the required temporal resolution, and we here use such data to compute stability properties as easy to interpret univariate indicators of microbial community dynamics. Such monitoring tools will allow for a fast, continuous, and cost-effective screening of stability states of microbiomes. Applicable to various environments, including bioreactors, surface water, and the human body, it will contribute to the development of control schemes to manipulate microbial community structures and performances.


Murine models of idiopathic inflammatory myopathy.
Idiopathic inflammatory myopathies (IIMs) are characterized by inflammation of muscles and other organs. Several myositis-specific autoantibodies (MSAs) have been identified in IIMs and were found to be associated with distinct clinical features. Although MSAs are valuable for the diagnosis of IIMs, the pathogenic roles of these antibodies remain unknown. To investigate the pathogenesis of IIMs, several animal models of experimental myositis have been established. Classical murine models of autoimmune myositis, experimental autoimmune myositis, and C protein-induced myositis are established by immunization with muscle-specific antigens, myosin, and skeletal C protein, respectively. Furthermore, a murine model of experimental myositis was generated by immunization with a murine recombinant histidyl-tRNA synthetase, Jo-1, in which muscle and lung inflammation reflecting anti-synthetase syndrome are induced depending on acquired immunity. Recently, the transfer of human IgGs from patients with immune-mediated necrotizing myopathy, comprising anti-signal recognition particles and anti-3-hydroxy-3-methylglutaryl coenzyme A reductase antibodies, was found to induce complement-mediated myositis in recipient mice. CD8+ T cell-mediated myositis can be established depending on autoimmunity against transcriptional intermediary factor 1γ (TIF1γ), an autoantigen for MSAs induced by recombinant human TIF1γ immunization. These new murine models reflecting MSA-related IIMs are useful tools for accurately understanding the pathological mechanisms underlying IIMs.