Plasmid DNA purification.
The demand for efficient production methods of plasmid DNA (pDNA) has increased vastly in response to rapid advances in the use of pDNA in gene therapy and in vaccines since the advantageous safety concerns associated with non-viral over viral vectors.A prerequisite for the success of plasmid-based therapies is the development of cost-effective and generic production processes of pDNA. However, to satisfy strict regulatory guidelines, the material must be available as highly purified, homogeneous preparations of supercoiled circular covalently closed (ccc) pDNA. Large-scale production of pDNA for therapeutic use is a relatively new field in bioprocessing. The shift from small-scale plasmid production for cell transfection to large-scale production sets new constraints on the bacterial fermentation, processing of bacterial lysate and final purification and formulation of the plasmid DNA. The choice of bacterial strain used for plasmid cultivation affects the plasmid yield, the proportion of different isoforms and the amount of endotoxins in the starting material. The choice of bacterial strain will be greatly influenced by the production and purification procedures of pDNA. Master and working cell banks need to be characterised and established. Alkaline lysis of the bacteria damages the pDNA, resulting in a reduced recovery of ccc pDNA and an increase in partially denaturated ccc pDNA and open circular (oc) forms. Shear stress in these processes needs to be tightly controlled, and buffer composition and pH need to be optimised. To obtain a homogeneous plasmid DNA preparation, different pDNA purification strategies aim at capturing ccc pDNA and eliminating the oc isoform. A highly purified final product corresponding to the stringent recommendations set forth by health and regulatory authorities can be achieved by (i). different chromatography techniques integrated with ultra/diafiltration to achieve optimal purification results; (ii). the formulation of the final pDNA product, that requires a detailed study of the plasmid structure; and (iii). the development of sensitive analytical methods to detect different impurities (proteins, RNA, chromosomal DNA, and endotoxins). We present here a revue of the whole process to obtain such a plasmid DNA, and report an example of RNAse-free purification of ccc pDNA that could be used for gene therapy.


Systematic Identification of Cell-Cell Communication Networks in the Developing Brain.
Since the generation of cell-type specific knockout models, the importance of inter-cellular communication between neural, vascular, and microglial cells during neural development has been increasingly appreciated. However, the extent of communication between these major cell populations remains to be systematically mapped. Here, we describe EMBRACE (embryonic brain cell extraction using FACS), a method to simultaneously isolate neural, mural, endothelial, and microglial cells to more than 94% purity in ∼4 h. Utilizing EMBRACE we isolate, transcriptionally analyze, and build a cell-cell communication map of the developing mouse brain. We identify 1,710 unique ligand-receptor interactions between neural, endothelial, mural, and microglial cells in silico and experimentally confirm the APOE-LDLR, APOE-LRP1, VTN-KDR, and LAMA4-ITGB1 interactions in the E14.5 brain. We provide our data via the searchable "Brain interactome explorer", available at https://mpi-ie.shinyapps.io/braininteractomeexplorer/. Together, this study provides a comprehensive map that reveals the richness of communication within the developing brain.


Pediatric Kidney Biopsies in Oman: A Retrospective Study.
To determine the spectrum of kidney diseases in Omani children < 13 years of age and to evaluate the complications following kidney biopsy. This study retrospectively investigated the hospital data of children who underwent kidney biopsies from January 2014 to June 2019 at Royal Hospital, Muscat, Oman. The subjects comprised of 78 children with a median age of 8.0 years (range = 0-13 years). Histopathology showed minimal change disease in 15 (19.2%) children, lupus nephritis in 13 (16.7%), and focal segmental glomerulosclerosis in 13 (16.7%). The most common post-biopsy complications were pain that required analgesia (38; 49.4%) followed by gross hematuria (10; 13.0%). No patient required blood transfusion or surgical intervention. Minimal change disease was the most common histopathological finding in this cohort of Omani children. The records did not mention any major complications following the renal biopsy procedure.


Heteroatom-substitution as a strategy for increasing the potency of competitive NMDA antagonists.
We report the synthesis and characterization of compounds that are competitive NMDA receptor antagonists. Significant increases in affinity and potency were obtained by incorporation of a heteroatom into the substructure of the tetrazole-substituted amino acid LY233053.


Endoscopic pancreatic drainage in chronic pancreatitis.
A nasopancreatic drain, pancreatic duct endoprostheses, and pancreatic stone extraction were used to treat 32 patients with chronic pancreatitis. Thirty patients were treated endoscopically. Endoscopic treatment via the minor papilla in 2 patients with pancreas divisum was not performed. Three patients had subsequent surgery because of complications; one of them died. Seventeen patients with chronic relapsing pancreatitis improved, with 15 patients asymptomatic during a follow-up of 2 to 69 months (median, 11). Seven of 10 patients with chronic pain improved, with 6 patients pain-free during a follow-up of 10 to 34 months (median, 11). In 7 patients, pancreatic pseudocysts could be drained endoscopically by positioning an endoprosthesis into the cyst or by performing a cystoduodenostomy. Six patients had concomitant placement of a biliary endoprosthesis to treat common bile duct strictures within the pancreatic head. One of 32 treated patients died as a result of a complication. We consider endoscopic therapy a viable alternative to surgery in select patients with chronic pancreatitis.


Carcinoma of the splenic flexure.
Carcinoma of the splenic flexure is uncommon and the diagnosis should be kept in mind, particularly in patients with recurring upper gastrointestinal symptoms. Resection is usually possible and operative complications are few. The site of the tumor does not affect long-term survival. Subtotal colectomy with ileosigmoid anastomosis would seem to be a safe method of treating patients with an obstructed carcinoma at that site.


The Extent of the Impact of a Fluorochemical Industrial Park in Eastern China on Adjacent Rural Areas.
High levels of perfluoroalkyl substances (PFASs) were observed in rural areas near a fluorochemical industrial park (FCIP) in the Yangtze River Delta region. The concentrations of total PFAS (∑PFASs) in soil, surface water, groundwater, and rainwater were in the range of 0.6-64.6 ng/g dry weight, 15.6-480.9, 4.8-614.6, and 13.4-542.2 ng/L, respectively. PFASs in rainwater and groundwater were higher than in surface water. The influence of the FCIP on rural soils extended to a distance of approximately 4 km based on the changes in PFAS compositions. The impact on surface water is mainly through direct river flow connected with contaminated sources, which may extend farther than 7 km in the case of unrestricted flow but much shorter distance for unconnected rivers. The penetration of PFASs in groundwater aquifers can extend as far as ca. 6 km, while direct water flow is the main transport route for PFASs. The influence radius of the FCIP can be as far as 60 km due to atmospheric precipitation.


Poisson-Delaunay Mosaics of Order k.
The order-k Voronoi tessellation of a locally finite set X ⊆ R n decomposes R n into convex domains whose points have the same k nearest neighbors in X. Assuming X is a stationary Poisson point process, we give explicit formulas for the expected number and total area of faces of a given dimension per unit volume of space. We also develop a relaxed version of discrete Morse theory and generalize by counting only faces, for which the k nearest points in X are within a given distance threshold.


COVID-19 and the amplification of cardiovascular risk by psychological distress.
The COVID-19 pandemic has unleashed a tidal wave of psychological distress. Here, we discuss the biobehavioral mechanisms through which psychological distress amplifyies the adverse effects of SARS-CoV-2-infection on cardiovascular outcomes. We also examine how the stress of caring for patients with COVID-19 increases cardiovascular risk in healthcare workers.


Travelling for treatment; does distance and deprivation affect travel for intensity-modulated radiotherapy in the rural setting for head and neck cancer?
NHS England has commissioned intensity-modulated radiotherapy for head and neck cancers from Newcastle hospitals for patients in North Cumbria. This study assessed whether travel distances affected the decision to travel to Newcastle (to receive intensity-modulated radiotherapy) or Carlisle (to receive conformal radiotherapy). All patients for whom the multidisciplinary team recommended intensity-modulated radiotherapy between December 2013 and January 2016 were included. Index of multiple deprivation scores and travel distances were calculated. Patients were also asked why they chose their treating centre. Sixty-nine patients were included in this study. There were no significant differences in travel distance (p = 0.53) or index of multiple deprivation scores (p = 0.47) between patients opting for treatment in Carlisle or Newcastle. However, 29 of the 33 patients gave travel distance as their main reason for not travelling for treatment. Quantitatively, travel distance and deprivation does not impact on whether patients accept intensity-modulated radiotherapy. However, patients say distance is a major barrier for access. Future research should explore how to reduce this.


Stimulation of reparative osteogenesis after xenotransplantation of human prenatal mesenchymal stem cells and chondroblasts.
Regeneration of the bone tissue after unilateral xenogeneic transplantation of a suspension of human mesenchymal cells and chondroblasts was studied in rats with experimental damage to both femurs. The state of animals was satisfactory and non-depressed in the early and late postoperation period. No local pathological reactions and complications were seen. Administration of mesenchymal stem cells and chondroblasts into the area of bone defect accelerated regeneration on days 10 and 30 (compared to the control group). Implantation of mesenchymal stem cells more significantly stimulated reparative osteogenesis compared to treatment with chondroblasts. This was mainly associated with increased ratio of mature lamellar bone tissue. Unilateral transplantation of the cell suspension stimulated regeneration in the contralateral limb due to accelerated maturation of the bone tissue. The bone tissue formed after transplantation of mesenchymal stem cells and chondroblasts was integrated into bone organ and underwent complete remodeling. Xenotransplantation of prenatal mesenchymal stem cells and chondroblasts without immunosuppression was not followed by the development of early and delayed complications or local reactions of graft rejection.


Consultant report: Incomplete abortions treated at Jahanshah Saleh Hospital in Tehran, Iran from May 14, 1973 to April 30, 1974.
In the period from May, 1973 to April, 1974, one hundred patients were treated for incomplete abortion at the Jahanshah Saleh Hospital in Tehran, Iran. Patients admitted to the hospital were from 4 to 28 weeks' gestation and were routinely administered analgestics before the abortion was completed by D & C. The D & C took about 10 minutes and the patient was usually able to go home that same day. Twenty-four women (24.0%) experienced complication(s) including blood loss, fever requiring antibiotic treatment, and pelvic infection.


Observed versus reported precepting skills: teaching behaviours in a community ambulatory clerkship.
Community-based doctors are increasingly utilised for the clinical education of medical students. Faculty development programmes are frequently provided. However, data about the community faculty-student teaching interaction and the longterm impact of such programmes are limited. This study observes community faculty-student teaching interactions and assesses the use of clinical teaching methods taught during faculty development workshops. Between March and August 2002, 13 rural, urban and suburban community-based faculty who completed at least 1 faculty development workshop were observed during faculty-student-patient encounters and interviewed. Observer-interviewers utilised a checklist to record teaching styles, methods and skills, and administered a questionnaire to obtain community faculty's self-perceptions. On observation, the assertive precepting style and didactic teaching predominated in 90% and 86% of teaching encounters, respectively. Interactive discussion and direct observation occurred less often, in 71% and 18% of encounters, respectively. Feedback was provided in 25% of encounters. On interview, community faculty reported that faculty development workshops resulted in improved precepting effectiveness and professional satisfaction with moderate to considerable improvement in feedback, interactive teaching and use of multiple precepting styles for 100%, 77% and 77% of community faculty, respectively. Community faculty reported that their skills in addressing problem students, direct observation and didactic teaching improved moderately. There was little statistical correlation between self-assessed and observed skills. Community-based faculty rate their teaching skills as improved following faculty development. Observation reveals that interactive discussion, direct observation, feedback and problem management skills are still lacking.


Suitability of the predatory mites Iphiseiodes zuluagai and Euseius concordis in controlling Polyphagotarsonemus latus and Tetranychus bastosi on Jatropha curcas plants in Brazil.
One of the most promising plant species for biofuel production in Brazil is the physic nut Jatropha curcas. Major phytosanitary problems include the attack of two pest mite species, the broad mite Polyphagotarsonemus latus and the spider mite Tetranychus bastosi. Owing to pesticide-related problems, there is an increasing demand for sustainable environmental-friendly control methods such as biological control. In this study we evaluated the suitability of the predatory mite species Iphiseiodes zuluagai and Euseius concordis in controlling P. latus and T. bastosi on J. curcas. The number of T. bastosi killed by I. zuluagai was lower than the number of P. latus consumed. Euseius concordis preyed upon both T. bastosi and P. latus but the number of prey killed was always lower in comparison with I. zuluagai. However, P. latus and T. bastosi are suitable for the development of I. zuluagai and E. concordis as oviposition of both predators did not differ in relation to prey species. The preference of I. zuluagai for leaves of plants infested by either P. latus or T. bastosi, combined with the higher values for predation obtained by this predatory mite when fed on P. latus, compared to those values obtained by E. concordis, suggests that I. zuluagai can be more efficient than E. concordis in reducing populations of P. latus and T. bastosi under field conditions. Furthermore, we report here on the first record of predatory mites associated with P. latus and T. bastosi on native J. curcas plants in Brazil. In conclusion, we emphasize the crucial importance of predatory mites as agents of natural biological control of mite pests on J. curcas in small farms.


A vigilance model for latent learning.
The author proposes a heuristic model for latent learning. It is concluded that to regard academic learning as qualitatively different from other forms of learning is to deny evolutionary continuity. Academic learning is not a unitary process governed by a single set of parameters. In addition, it is observed that the problem of student motivation may very well turn out to be purely academic. The instructional technique for a captive audience of a class may be so structured as to make the direction of attention irresistible, the performance of a response, when needed, compelling, and the acquisition of knowledge inevitable. Vigilance is an instance of innate foundation. Its most striking characteristics are its universality in the animal world, its ready evocation by a wide range of stimuli, and its apparent behavior and physiological manifestations. The last two are the natural resources for objective investigation, and the first may well be the basis of broad and valid generalizations.


Intracardiac echocardiography: gross anatomy and magnetic resonance correlations and validations.
The feasibility and safety of intracardiac echocardiography (ICE) in humans, using low frequency transducers, and its excellent tissue contrast capabilities that enhances the differentiation of intracardiac structures have been previously demonstrated. However, correlations among ICE imaging and anatomic sections or magnetic resonance (MR) scan planes have never been described before. This study was designed to correlate a simplified ICE approach with the anatomy of the right atrium and great vessels obtained by serial post-mortem sections and cardiac MR images. A stepwise approach to ICE, which is based on our experience on over 300 consecutive patients with interatrial communications, has been correlated with anatomic sections from pressure-perfused-fixed hearts and spin echo cardiac MR imaging. A 9F-9 MHz mechanical device was used to record four transverse and one longitudinal sections for an extensive evaluation of the intracardiac architecture. ICE transverse and longitudinal views allowed the detection of all the required information. Moreover, the anatomic sections and cardiac MR scan planes allowed validation of all the structures imaged by ICE. The potential clinical applications of the ICE technique are further discussed. This paper demonstrates the basic accuracy of this new imaging modality. Therefore, mechanical ICE might be considered an active investment in cardiac catheterization laboratories, specifically in the percutaneous interventional procedure setting, discovering a new route (let's see and treat) to sophisticated interventions. In this respect, there can be little doubt that the knowledge of cross-sectional mechanical ICE imaging provides the basis for understanding the heart anatomy.


The GLT-1 and GLAST glutamate transporters are expressed on morphologically distinct astrocytes and regulated by neuronal activity in primary hippocampal cocultures.
The GLT-1 and GLAST astroglial transporters are the glutamate transporters mainly involved in maintaining physiological extracellular glutamate concentrations. Defects in neurotransmitter glutamate transport may represent an important component of glutamate-induced neurodegenerative disorders (such as amyotrophic lateral sclerosis) and CNS insults (ischemia and epilepsy). We characterized the protein expression of GLT-1 and GLAST in primary astrocyte-neuron cocultures derived from rat hippocampal tissues during neuron differentiation/maturation. GLT-1 and GLAST are expressed by morphologically distinct glial fibrillary acidic protein-positive astrocytes, and their expression correlates with the status of neuron differentiation/maturation and activity. Up-regulation of the transporters paralleled the content of the synaptophysin synaptic vesicle marker p38, and down-regulation was a consequence of glutamate-induced neuronal death or the reduction of synaptic activity. Finally, soluble factors in neuronal-conditioned media prevented the down-regulation of the GLT-1 and GLAST proteins. Although other mechanisms may participate in regulating GLT-1 and GLAST in the CNS, our data indicate that soluble factors dependent on neuronal activity play a major regulating role in hippocampal cocultures.


Significant effect of size on the in vivo biodistribution of gold composite nanodevices in mouse tumor models.
There is growing interest in developing tissue-specific multifunctional drug delivery systems with the ability to diagnose or treat several diseases. One class of such agents, composite nanodevices (CNDs), is multifunctional nanomaterials with several potential medical uses, including cancer imaging and therapy. Nanosized metal-dendrimer CNDs consist of poly(amidoamine) dendrimers (in various sizes, surface substituents, and net charges) and inorganic nanoparticles, properties of both of which can be individually modified and optimized. In this study we examine effects of size and surface charge on the behavior of Au-dendrimer CNDs in mouse tumor models. Quantitative biodistribution and excretion analyses including 5-nm and 22-nm positive surface, 5-nm and 11-nm negative surface, and a 5-nm neutral surface CNDs were carried out in the B16 mouse melanoma tumor model system. Results seen with the 22-nm CND in the B16 melanoma model were corroborated in a prostate cancer mouse tumor model system. Quantitative in vivo studies confirm the importance of charge and show for the first time the importance of size in affecting CND biodistribution and excretion. Interestingly, CNDs of different size and/or surface charge had high levels of uptake ("selective targeting") to certain organs without specific targeting moieties placed on their surfaces. We conclude that size and charge greatly affect biodistribution of CNDs. These findings have significance for the design of all particle-based nanodevices for medical uses. The observed organ selectivity may make these nanodevices exciting for several targeted medical applications.


Isolated angiomyolipoma of vulva: A case report of an uncommon tumor at an uncommon site.
Extra renal angiomyolipoma is an exceedingly rare entity. This is usually not associated with tuberous sclerosis as it has no human melanoma black 45 immune reactive cells although it shares the terminology with renal angiomyolipoma. The histological features are characterized by admixture of blood vessels, smooth muscles, and mature adipose tissues in variable proportion. Surgical excision is considered curative. We report a case of isolated vulval angiomyolipoma in a 45-year-old female which clinically and radiologically presented as lipoma like lesion. Our case of isolated vulval angiomyolipoma is the first reported case in the literature.


What is the Optimal Frequency of Visual Field Testing to Detect Rapid Progression Among Hypertensive Eyes?
We evaluated 16,351 visual field (VF) tests from Ocular Hypertension Treatment Study (OHTS) database and showed that more frequent testing resulted in a shorter time to detect glaucoma progression, with the best trade-off being the 6-month intervals for high-risk and 12 months for low-risk patients. To investigate the effect of different testing intervals on time to detect visual field progression in eyes with ocular hypertension. A total of 16,351 reliable 30-2 VF tests from 1575 eyes of the OHTS-1 observation arm with a mean (95% CI) follow-up of 4.8 (4.7-4.8) years were analyzed. Computer simulations (n = 10,000 eyes) based on mean deviation values and the residuals of risk groups (according to their baseline 5 y risk of developing primary open angle glaucoma: low, medium, and high risk) were performed to estimate time to detect progression with testing intervals of 4, 6, 12, and 24 months using linear regression. The time to detect VF progression ( P < 5%) at 80% power was calculated based on the mean deviation slope of -0.42 dB/year. We assessed the time to detect a -3 dB loss as an estimate of clinically meaningful perimetric loss. At 80% power, based on the progression of -0.42 dB/year, the best trade-off to detect significant rates of VF change to clinically meaningful perimetric loss in high, medium, and low-risk patients was 6, 6, and 12-month intervals, respectively. Given the importance of not missing the conversion to glaucoma, the frequency of testing used in OHTS (6 mo) was optimal for the detection of progression in high-risk patients. Low-risk patients could potentially be tested every 12 months to optimize resource utilization.


Intronic Determinants Coordinate Charme lncRNA Nuclear Activity through the Interaction with MATR3 and PTBP1.
Chromatin architect of muscle expression (Charme) is a muscle-restricted long noncoding RNA (lncRNA) that plays an important role in myogenesis. Earlier evidence indicates that the nuclear Charme isoform, named pCharme, acts on the chromatin by assisting the formation of chromatin domains where myogenic transcription occurs. By combining RNA antisense purification (RAP) with mass spectrometry and loss-of-function analyses, we have now identified the proteins that assist these chromatin activities. These proteins-which include a sub-set of splicing regulators, principally PTBP1 and the multifunctional RNA/DNA binding protein MATR3-bind to sequences located within the alternatively spliced intron-1 to form nuclear aggregates. Consistent with the functional importance of pCharme interactome in vivo, a targeted deletion of the intron-1 by a CRISPR-Cas9 approach in mouse causes the release of pCharme from the chromatin and results in cardiac defects similar to what was observed upon knockout of the full-length transcript.


Immediate amputation stump coverage with forearm free flaps from the same limb.
When the traumatic amputation level or the degree of damage to the arm or forearm does not justify reimplantation of the severed part there may be sufficient undamaged skin to be used as a free flap to resurface the arm or the stump. Stump length or the elbow joint may thus be preserved and more durable skin cover for the fitting of a prosthesis may be obtained. A "spare-parts" forearm free flap may be raised on the radial or ulnar vessels or both. Three cases are reported.


Possible application of CT morphometry of the calcaneus and talus in forensic anthropological identification.
Computed tomography (CT) data provide information for volumetric and radiographic density analysis. The present study investigated the application of virtual CT volumetry of the tarsal bones to estimation of the sex, stature, and body weight using postmortem CT (PMCT) data of forensic autopsy cases. Three-dimensional (3D) images of the bilateral foot bones of intact Japanese subjects after adolescence (age ≥ 15 years, n = 179, 100 males and 79 females) were reconstructed on an automated CT image analyzer system. Measured parameters were mass volume, mean CT value (HU), and total CT value of the talus and calcaneus. Mean CT values of these bones showed age-dependent decreases in elderly subjects over 60 years of age for both sexes, with significant sex-related differences especially in the elderly. The mass volumes and total CT values of the talus and calcaneus showed significant sex-related differences, and also moderate correlations with body height and weight for bilateral bones in all cases (r = 0.58-0.78, p < 0.0001); however, the correlations of these parameters of the female talus with body weight were insufficient (r = 0.41-0.61, p < 0.0001). These observations indicate the applicability of virtual CT morphometry of the talus and calcaneus using an automated analyzer to estimate the sex and stature in forensic identification; however, greater variations should be considered in body weight estimations of females.


Fabrication of porous bovine pericardium scaffolds incorporated with bFGF for tissue engineering applications.
The design and fabrication of porous scaffolds are important issues for tissue engineering applications. In this study, we attempted to fabricate porous scaffolds using bovine pericardium (BP) and examined whether these scaffolds were beneficial for cell ingrowth and bioactive factors delivery. A vacuum-freeze-thawing-Triton X-100 (VFTT) protocol was used to fabricate porous BP scaffolds. The porous and mechanical properties were assessed using histology, scanning electron microscopy, and mechanical assay. The fabricated scaffolds were seeded with mesenchymal stem cells (MSCs), and cell ingrowth was evaluated. Basic fibroblast growth factor (bFGF) was subsequently incorporated into the fabricated scaffolds. The bioactive factor delivery capacity was evaluated using loading and release studies. The bioactivity of released bFGF was assessed using a rat subcutaneous model. The BP scaffolds fabricated by the VFTT protocol displayed interconnected porous structures with porosity of 6.82 ± 1.36%.There were no significant differences in thickness, ultimate load, Young's modulus, and ultimate tensile strength between the fabricated porous BP scaffolds and native BPs (all P > .05). However, the water content of BPs was slightly reduced after VFTT treatment (P < .05). Cell ingrowth analysis showed that the seeded MSCs penetrated into the porous BP scaffolds with time of culture, while MSCs were limited to the surface layers of native BPs. Furthermore, bFGF was observed to be effectively loaded onto and released from the porous BP scaffolds. The released bFGF increased the phosphorylation levels of Akt, ERK 1/2, and MEK1/2, promoted host MSC recruitment, and inhibited myofibroblast differentiation in vivo. The porous BP scaffolds fabricated using a VFTT protocol were promising natural scaffolds for tissue engineering applications, since they had considerable mechanical properties as native BPs, supplied porous channels for cell ingrowth, and possessed bioactive factors delivery capability.


Impact of aluminum adjuvants on the stability of pneumococcal polysaccharide-protein conjugate vaccines.
Development of a vaccine drug product requires formulation optimization to ensure that the vaccine's effectiveness is preserved upon storage throughout the shelf-life of the product. Although aluminum adjuvants have been widely used in vaccine formulations to safely and effectively potentiate an immune response, careful attention must be directed towards ensuring that the type of aluminum adjuvant does not impact the stability of the antigenic composition. PCV15 is a polysaccharide-protein conjugate vaccine comprising the pneumococcal polysaccharide (PnPs) serotypes (1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, 22F, 23F and 33F), each individually conjugated to the protein carrier CRM197. PCV15 was formulated with either amorphous aluminum hydroxyphosphate sulfate adjuvant (AAHS) or aluminum phosphate adjuvant (AP) and examined for both stability and immunogenicity. Using a collection of methods to evaluate vaccine stability, it was discovered that certain PCV15 serotypes (e.g., 6A, 19A, 19F) formulated with AAHS resulted in a reduction of immunogenicity in vivo and a reduction in recoverable dose as tested by an in vitro potency assay. The same polysaccharide-protein conjugates formulated with AP were stable regarding all measures tested. Moreover, the reduction in potency of certain serotypes correlated with chemical degradation of the polysaccharide antigen caused by the aluminum adjuvant as measured by reducing polyacrylamide gel electrophoresis (SDS-PAGE), High-Pressure Size Exclusion Chromatography coupled with UV detection (HPSEC-UV) and ELISA immunoassay. This study suggests a formulation, which includes AAHS, may negatively impact the stability of a pneumococcal polysaccharide-protein conjugate vaccine that contains phosphodiester groups. This decrease in stability would likely result in a decrease in the "active" concentration of antigen dose, and herein, it is shown that such instability directly compromised vaccine immunogenicity in an animal model. The results presented in this study help to explain critical degradation mechanisms of pneumococcal polysaccharide-protein conjugate vaccines.


The impact of pre-registration education on the motivation and preparation of midwifery students to work in continuity of midwifery care: An integrative review.
Access to continuity of midwifery care (CoMC) models in Australia is increasing but the capacity of the emerging midwifery workforce to provide this care remains largely unknown. The aim of this integrative literature review is to discover how well pre-registration midwifery education prepares and motivates Australian midwifery students to work in CoMC models when they enter practice. Following title review of 432 papers, removal of duplicates and review against the inclusion and exclusion criteria, nine papers were included for review. The results show that access to CoMC is a crucial component of midwifery education, equipping students with knowledge, skills, confidence and motivation to work in this way upon graduation. Existing methods of program delivery and institutional structures often present students with challenges that detract from the value of their CoMC experiences. A focus on CoMC placement - particularly with a continuity of midwifery mentor - may motivate graduates to work in this model of care. This strategy is recommended to better align Australian midwifery education with maternity care reform.


The impact of industrial structure adjustment on the spatial industrial linkage of carbon emission: From the perspective of climate change mitigation.
Industrial structure adjustment is an effective way to reduce carbon emissions. When adjusting the industrial structure, changes in the input and output of industrial sectors in one region may cause changes in industrial sectors of other regions, resulting in inter-regional industrial linkages. What is the impact of industrial structure adjustment on the linkages, and how does it impact climate change? The modified gravity model combines the influence and inductive coefficients to build the carbon emission industrial spatial linkage network (ISL) using social network analysis (SNA) in this work with the data of China's 2008-2019 provincial carbon emissions. The impact of industrial structure adjustment on carbon emission industrial spatial linkage network and its effect on climate change mitigation are explored with the help of the STIRPAT model and quality assurance procedure (QAP). The results of this study are as follows: (1) The provincial carbon emissions under industrial spatial linkages show significant network characteristics, the network density increased from 0.1437 in 2008 to 0.323 in 2019. (2) There is a significant spatial spillover effect in the industrial spatial linkage network (2019) centered on Shandong and Jiangsu. The provinces can be divided into 4 blocks, which play different roles in the network. (3) The industrial structure upgrading can significantly promote the carbon emission industrial linkage network and mitigate climate change. (4) The industrial structure rationalization can promote the development of the network and mitigate climate change in the early and late development stages of carbon emissions industrial linkage. In addition, it is necessary for the government to pay more attention to the development level of the carbon emission spatial industrial linkage network when implementing industrial structure adjustment in response to climate change.


Apparent microviscosity of intact and post-lipolysis ("remnant") very low density lipoprotein particles.
The apparent microviscosity of intact rat plasma very low density lipoprotein (VLDL) and post-lipolysis very low density lipoprotein was determined by fluorescence depolarization measurements and flurorescence decay measurements using 1, 6-diphenylhexatriene. Post-lipolysis very low density lipoprotein was prepared in vitro after incubation of the intact lipoprotein with either purified bovine milk lipoprotein lipase or lipoprotein lipase rich (post-heparin) plasma. During lipolysis, an average of 88% of the triglycerides were hydrolyzed, and the lipoprotein became depleted in phospholipids, cholesterol and apolipoprotein C. The apparent microviscosity of the lipoprotein increased by three-fold from 0.63 to 1.88 poise. It is concluded that the compositional changes occurring during lipolysis affect the physical properties of the lipoprotein, as measured here by the fluidity (microviscosity) of the particles.


Effects of non-sporting and sporting qigong on frailty and quality of life among breast cancer patients receiving chemotherapy.
To explore the effects of non-sporting qigong (NSQG) and sporting qigong (SQG) on frailty and quality of life (QOL) of breast cancer patients during chemotherapy. A time series (three-group, pre-test-post-test) quasi-experimental design was applied in the study. Ninety-five participants were assigned to three groups: controls (n = 31), NSQG (n = 33), or SQG (n = 31). All patients performed the qigong interventions three times per week for at least 30 min per session. Data were collected in face-to-face interviews before chemotherapy and at 1 and 3 months after chemotherapy. Frailty was assessed using the Edmonton Frail Scale. The Medical Outcomes Survey Short-Form 36-Taiwanese version was used to evaluate the physical and mental component scores of QOL. In the 1st and 3rd months after practicing qigong, patients in the SQG group had lower frailty scores than those in the control group. In the 3rd month after the intervention, patients in the NSQG group also had lower frailty scores and higher mental component scores for QOL than those in the control group. Patients with higher frailty scores had worse physical and mental component scores for QOL than those with lower frailty scores. The Sobel test showed that the frailty score mediated SQG and physical component scores for QOL. SQG and NSQG appeared to be beneficial for improving frailty and QOL among the breast cancer patients receiving chemotherapy in the study. The results are preliminary and larger, well-constructed clinical studies are needed to verify the findings.


DNA Cytometry and Nuclear Morphometry in Ovarian Benign, Borderline and Malignant Tumors.
Ovarian carcinoma is a leading cause of death in gynecological malignancy. Ovarian surface epithelial serous and mucinous tumours are classified as benign, borderline, and malignant. The identification of borderline tumours most likely to act aggressively remains an important clinical issue. This work aimed to study DNA ploidy and nuclear area in ovarian serous and mucinous; benign, borderline and malignant tumours. This study included forty ovarian (23 serous and 17 mucinous) tumours. Paraffin blocks were sectioned; stained with haematoxylin and eosin for histopathologic and morphometric studies and with blue feulgen for DNA analysis. All four serous and six out of nine mucinous benign tumours were diploid. All eight serous and five mucinous malignant tumours were aneuploid. Nine of eleven (81.8%) serous and all three mucinous borderline tumours were aneuploid. There were highly significant differences in mean aneuploid cells percentage between serous benign (1.5%), borderline (45.6%) and malignant (74.5%) (p = 0.0001) and between mucinous benign (13.2%) and both borderline (63.7%) and malignant (68.4%) groups (p = 0.0001). There were significant differences in nuclear area between serous benign (26.191%), borderline (45.619%) and malignant (67.634 %) and a significant positive correlation between mean percentage aneuploid value and mean nuclear area in all serous and mucinous groups. We suggest that DNA ploidy and nuclear area combined, may be adjuncts to histopathology; in ovarian serous and mucinous benign, borderline and malignant neoplasms; identifying the aggressive borderline tumours.


Progression of articular destruction and the production of tumour necrosis factor-alpha in antigen-induced arthritis in rabbits.
We examined the progression of articular destruction and the production of tumour necrosis factor-alpha (TNF-alpha) in antigen-induced arthritis (AIA) in rabbits, i.e. flare-ups of inflammation induced by repeated intra-articular injections (single, twice and three times) of antigen. A marked progression of articular destruction and an infiltration of inflammatory cells in the synovium were observed with the increase in the number of antigen injections. An immunohistochemical analysis of the synovial lesions following three injections of antigen revealed that the lymphoid follicles consisted mainly of CD4+ T cells and IgG/IgM+ B cells. There were marked infiltrations of IgG+ plasma cells around the lymphoid follicles. In contrast, the production of TNF-alpha in the synovial fluid and the erythrocyte sedimentation rate (ESR), which is a marker of systemic inflammatory activity in rheumatoid arthritis, peaked at 6 h 24 h, respectively, following the last injection of antigen. These values were also greater following the repeated injections of antigen compared with the single injection. The TNF-alpha was produced markedly in the joints at the onset of the flare-ups of arthritis following the repeated injections of antigen, and the elevation of the ESR and an acceleration of the inflammatory response in the synovium were observed with a concomitant progression of severe articular destruction, suggesting that the marked production of TNF-alpha at the time of flare-ups may be involved in the exacerbation of AIA in rabbits.


Functionalized NU-1000 with an Iridium Organometallic Fragment: SO2 Capture Enhancement.
A new material, MOF-type [Ir]@NU-1000, was accessed from the incorporation of the iridium organometallic fragment [Ir{κ3(P,Si,Si)PhP(o-C6H4CH2SiiPr2)2}] into NU-1000. The new material incorporates less than 1 wt % of Ir(III) (molar ratio Ir to NU-1000, 1:11), but the heat of adsorption for SO2 is significantly enhanced with respect to that of NU-1000. Being a highly promising adsorbent for SO2 capture, [Ir]@NU-1000 combines exceptional SO2 uptake at room temperature and outstanding cyclability. Additionally, it is stable and can be regenerated after SO2 desorption at low temperature.


[Prevention and management of pegaspargase associated-toxicities (excluding coagulation abnormalities). Recommendations of the French Society of Children and Adolescent Cancers (Leukemia committee)].
Pegaspargase (Oncaspar®), a pegylated form of native Escherichia Coli-derived L-asparaginase is an essential component chemotherapy used in the treatment of acute lymphoblastic leukemia (ALL) in pediatric and adult patients. Its particular toxicity profile requires a specific management to improve safety and tolerability and optimize treatment outcome and therefore survival. Within the framework of workshops of practice harmonization of the French Society of Children and Adolescent Cancers, diagnostic and management of the most commonly occuring toxicities (excluding coagulation abnormalities) during Pegaspargase treatment were reviewed according to the analysis of published studies.


Comparative pharmacokinetics of six bioactive components of Shen-Wu-Yi-Shen tablets in normal and chronic renal failure rats based on UPLC-TSQ-MS/MS.
Shen-Wu-Yi-Shen tablets (SWYST), a Chinese patent medicine consisting of 12 herbal medicines, was formulated by a famous TCM nephrologist, Zou Yunxiang. It is clinically used to improve the symptoms of nausea, vomiting, poor appetite, dry mouth and throat, and dry stool in patients with chronic renal failure (CRF) accompanied by qi and yin deficiency, dampness, and turbidity. SWYST can reduce urea nitrogen, blood creatinine, and urinary protein loss, and increase the endogenous creatinine clearance rate. However, little is known about its pharmacokinetics. To compare the pharmacokinetics of six bioactive components after oral administration of SWYST in normal and adenine-induced CRF rats. A method based on ultra-performance liquid chromatography coupled with a triple-stage quadrupole mass spectrometer (UPLC-TSQ-MS/MS) was developed and validated to determine the six bioactive compounds (albiflorin, paeoniflorin, plantagoguanidinic acid, rhein, aloe-emodin, and emodin) in rat plasma. Rat plasma samples were prepared using protein precipitation. Chromatography was performed on an Agilent Eclipse Plus C18 column (3.0 × 50 mm, 1.8 μm) using gradient elution with a mobile phase composed of acetonitrile and water containing 0.1% (v/v) formic acid, while detection was achieved by electrospray ionization MS under the multiple selective reaction monitoring modes. After SWYST administration, rat plasma was collected at different time points, and the pharmacokinetic parameters of six analytes were calculated and analyzed based on the measured plasma concentrations. The UPLC-TSQ-MS/MS method was fully validated for its satisfactory linearity (r ≥ 0.9913), good precisions (RSD <11.5%), and accuracy (RE: -13.4∼13.1%), as well as acceptable limits in the extraction recoveries, matrix effects, and stability (RSD <15%). In normal rats, the six analytes were rapidly absorbed (Tmax ≤ 2 h), and approximately 80% of their total exposure was eliminated within 10 h. Moreover, in normal rats, the AUC0-t and Cmax of albiflorin, plantagoguanidinic acid, and rhein exhibited linear pharmacokinetics within the dose ranges, while that of paeoniflorin is non-linear. However, in CRF rats, the six analytes exhibited reduced elimination and significantly different AUC or Cmax values. These changes may reflect a decreased renal clearance rate or inhibition of drug-metabolizing enzymes and transporters in the liver and gastrointestinal tract caused by CRF. A sensitive UPLC-TSQ-MS/MS method was validated and used to investigate the pharmacokinetics of SWYST in normal and CRF rats. This is the first study to investigate the pharmacokinetics of SWYST, and our findings elucidate the causes of their different pharmacokinetic behaviors in CRF rats. Furthermore, the results provide useful information to guide further research on the pharmacokinetic-pharmacodynamic correlation and clinical application of SWYST.


Screening for prolonged incubation of HTLV-I infection in British and Jamaican relatives of British patients with tropical spastic paraparesis.
To compare the prevalence of antibody to and proviral DNA of the retrovirus HTLV-I in relatives of 11 British patients with tropical spastic paraparesis who had migrated from Jamaica before they developed symptoms, and to examine factors possibly related to transmission of HTLV-I. Migrant, family study. Antibody state was determined by several methods and confirmed by western blotting; the polymerase chain reaction was used to detect proviral DNA. Britain and Jamaica. All available first degree relatives: those born and still resident in Jamaica (group 1); those born in Jamaica who migrated to Britain (group 2); and index patients' children who were born and resident in Britain (group 3). All had been breast fed and none had had blood transfusions. Of the 66 living relatives, 60 were traced. Seroprevalence among those born in Jamaica (irrespective of current residence) was 22% (10/46; 95% confidence limits 9 to 34%) compared with zero among British born offspring (0/14) and was higher in group 2 at 33% (7/21; 12 to 55%) than in group 1 at 12% (3/25; 0 to 25%). (Patients in group 1 had the greatest mean age.) Proviral DNA was not detected in any subject negative for HTLV-I antibody, making prolonged viral incubation in those negative for the antibody unlikely. In this sample factors related to place of birth and early residence were more important in transmission of HTLV-I than maternal or age effects. In areas with a low to moderate prevalence policies of preventing mothers who are carriers of the virus from breast feeding would be premature.


[The Werther effect. Significance, mechanisms, consequences].
The Werther effect is a highly regarded phenomenon in media effect research. Since the trail-blazing studies conducted by the American sociologist Phillips, there have been a great many attempts to quantify and evaluate imitative deeds that follow genuine suicides and apparent attempts at suicide. This paper starts by tracing the milestones along the way of research into the Werther effect. The concrete imitation patterns following suicides are then discussed, and the implications for responsible journalistic reporting that arise out of them are presented. This seems to be necessary, since despite many-sided and detailed findings on the Werther effect in Germany, there have so far been no experiments devoted to ways of exploiting these research results practically in a suicide prevention programme. Whilst in other countries preliminary guidelines for journalistic reporting have already been elaborated and in some countries even implemented, such efforts are still only in the very early stages in Germany.


Purification and characterization of chicken brain cytosolic aspartate aminotransferase.
Aspartate aminotransferase from the cytosolic fraction of chicken brain was isolated with acceptable yield and high degree of purity. The enzyme appeared in multiple molecular forms: alpha, beta, gamma, and delta (alpha predominates), as detected by polyacrylamide gel electrophoresis with specific staining. These different forms of the enzyme were separated by DEAE-Sephacel chromatography, and showed different isoelectric points and maximal velocities values, whereas their molecular weight, optimum pH and Michaelis constants were very similar. Generation process studies suggest that minors subforms of the enzyme could be raised from alpha form by a mechanism in which the oxidation of particular amino acid groups are involved.


Laparoscopic transhiatal esophagectomy for Barrett's esophagus with high grade dysplasia.
A number of case reports have described the application of minimally invasive surgical techniques to accomplish esophagectomy. However, most reports have employed thoracoscopic or laparoscopic techniques to perform esophagectomy in addition to an "access" incision which often approaches a standard laparotomy or thoracotomy. This report describes a total laparoscopic transhiatal esophagectomy in a 55 year old female with Barrett's esophagus and high grade dysplasia. The patient had an uneventful recovery and was discharged home on the fourth day after a total laparoscopic esophagectomy. This report demonstrates the technical feasibility of this complex procedure by a minimally invasive approach.


Inhibition of pyrimidine and purine nucleoside phosphorylases by a 3,5-dichlorobenzoyl-substituted 2-deoxy-D-ribose-1-phosphate derivative.
The 3,5-dichlorobenzoyl-substituted 2-deoxy-D-ribose-1-phosphate derivative, designated Cf2891, was found to inhibit a variety of pyrimidine and purine nucleoside phosphorylases (NPs) with preference for uridine- and inosine-hydrolyzing enzymes [uridine phosphorylase (UP; EC 126.96.36.199), pyrimidine nucleoside phosphorylase (PyNP; EC 188.8.131.52) and purine nucleoside phosphorylase (PNP; EC 220.127.116.11)]. Kinetic analyses revealed that Cf2891 competes with inorganic phosphate (P(i)) for binding to the NPs and, depending on the nature of the enzyme, acts as a competitive or non-competitive inhibitor with regard to the nucleoside binding site. Also, the compound prevents breakdown of pyrimidine analogues used in the treatment of viral infections and cancer. Since NPs are abundantly present in tumor tissue and may be overexpressed due to secondary bacterial infections in immunocompromised patients suffering viral infections, Cf2891 may serve as a lead molecule for the development of inhibitors to be used in nucleoside-based combination therapy.


Introduced reindeer and their effects on the vegetation and the epigeic invertebrate fauna of South Georgia (subantarctic).
The introduced reindeer of South Georgia have had a serious impact on the vegetation throughout the range of three populations on the island. Exclosure experiments in areas where the reindeer were introduced have resulted in a dramatic change in the composition of the protected vegetation. Poa flabellata (the major winter food) and Acaena magellanica (a major summer food) have recovered to their former status inside the exclosures, while Deschampsia antarctia and the introduced grass Poa annua tolerate grazing and trampling and have spread over the grazed areas. Festuca contracta and Rostkovia magellanica are not eaten by the reindeer. Trampling has resulted in a high proportion of bare soil and peat in sites freely accessible to reindeer. However, the changes in the vegetation have not had such a significant effect on the associated invertebrate fauna. Thus in grazed and protected areas the faunistic composition is qualitatively similar, although there are quantitative differences, and some of the trends can probably be attributed to the presence of the reindeer. Compared with reindeer-free areas, the abundance of the perimylopid beetle Hydromedion sparsutum (a major primary decomposer) is reduced. The frequency of their egg parasite Notomymar aptenosoma (Hymenoptera, Mymaridae) increases. Consequently the ratio of perimylopoids to mymarids found in pitfall traps shifts from 1:0.01 (ungrazed areas) to 1:0.54 (grazed areas). Also the frequency of sciarids was found to be higher in reindeer areas. The larvae of these probably introduced gnats do not play a role in the natural terrestrial ecosystem of South Georgia, but in reindeer areas they appear to establish larger populations because they are able to live deeper in the soil and in hardened substrates. There is also a shift in the ratio between Collembola (major prey) to spiders from 1:1.3 (ungrazed areas) to 1:0.82 (grazed areas), for animals collected in pitfall traps. This may be a result of the trampling effect of the reindeer.


Management of Conductive Hearing Loss in Children.
Conductive hearing loss (CHL), far more common than sensorineural hearing loss in children, can be acquired or congenital, can range from mild to moderately severe, and can be caused by a simple cerumen impaction, middle ear fluid, or complex middle ear abnormalities with or without the absence of the ear canal (congenital aural atresia). This article presents evidence-based recommendations for the evaluation and management of the child with both acquired and congenital CHL.


Nonsteroidal anti-inflammatory analgesics. Indications and contraindications for pain management in dogs and cats.
Nonsteroidal anti-inflammatory analgesics (NSAIAs) are effective in controlling most acute and chronic pain conditions. In veterinary practice these analgesics may be superior to opioids in that the duration of action is much longer, with equal efficacy in many instances, making effective pain management possible for most veterinary patients. NSAIAs act synergistically in combination with other modalities of pain management, including all opioids, local anesthetics, and various sedatives. Because of their mechanism of action, however, there is a potential for perturbation of several homeostatic functions mediated by prostaglandins. Not all NSAIAs are equal in efficacy and safety, so careful patient and NSAIA selection with appropriate monitoring is advised. This article discusses the indications and contraindications for NSAIA use with a short description of the currently available NSAIAs approved for use in veterinary patients.


Putative protein markers in the sera of men with prostatic neoplasms.
To describe the preliminary identification of serum proteins that may be diagnostic markers in prostate cancer. The study included 11 men referred for treatment of localized prostate cancer, 12 with benign prostatic hyperplasia (BPH) and 12 disease-free controls. For serum protein analysis, the protein-chip array surface-enhanced laser desorption/ionization (SELDI) technique was used (Ciphergen Biosystems, Fremont, CA). SELDI combines protein-chip technology with time-of-flight mass spectrometry, and offers the advantages of speed, simplicity and sensitivity. Three protein peaks were identified in the serum of men with prostate cancer and BPH, but not in controls, with relative molecular masses of 15.2, 15.9 and 17.5 kDa. These three proteins were significantly associated with BPH and prostate cancer when compared with controls (P = 0.001, 0.004, and 0.011, respectively, Kruskal-Wallis test). Interestingly, the 17.5 kDa protein was more abundant in five men with stage T1 prostate cancer than in eight with stage T2 (P = 0.016, two tailed Mann-Whitney U-test corrected for ties). These proteins, particularly the 15.9 kDa one, may be used for the diagnosis or monitoring of prostate cancer and differentiation from BPH, and have the potential for antibody-based chip SELDI-TOF technology. Identified proteins may be targets for immunotherapy.


Astragaloside IV and cycloastragenol are equally effective in inhibition of endoplasmic reticulum stress-associated TXNIP/NLRP3 inflammasome activation in the endothelium.
Astragaloside IV and cycloastragenol are present together in Astragalus membranaceus Moench (Fabaceae) and this study aims to simultaneously investigate their regulation of endothelial homeostasis in the setting of endoplasmic reticulum stress (ER stress). We stimulated endothelial cells with palmitate (PA 100μM) to evoked ROS-associated ER stress and observed the effects of astragaloside IV and cycloastragenol on thioredoxin-interacting protein (TXNIP) expression, NLRP3 inflammasome activation and mitochondrion-dependent apoptosis. Astragaloside IV and cycloastragenol inhibited ROS generation and attenuated ER stress inducer IRE1α phosphorylation, indicating the inhibition of ROS-associated ER stress. In response to ER stress, TXNIP expression increased, accompanied with NLRP3 induction and increased IL-1β and IL-6 production, but these alternations were reversed by treatment with astragaloside IV and cycloastragenol, demonstrating the inhibitory effects of astragaloside IV and cycloastragenol on TXNIP/NLRP3 inflammasome activation. Inflammasome activation led to mitochondrial cell death in endothelial cells, whereas astragaloside IV and cycloastragenol restored the loss of the mitochondrial membrane potential with inhibition of caspase-3 activity, and thereby protected cells from ER stress-induced apoptosis. Astragaloside IV and cycloastragenol enhanced AMPK phosphorylation and AMPK inhibitor compound C diminished their beneficial effects, indicative of the potential role of AMPK in their regulation. Astragaloside IV and cycloastragenol suppressed ROS-associated ER stress and then inhibited TXNIP/NLRP3 inflammasome activation with regulation of AMPK activity, and thereby ameliorated endothelial dysfunction by inhibiting inflammation and reducing cell apoptosis. Simultaneous investigations further showed that astragaloside IV and cycloastragenol were equally effective in regulation of endothelial homeostasis.


Carotenoid profile and retention in yellow-, purple- and red-fleshed potatoes after thermal processing.
This research aimed to investigate the effect of thermal processing on carotenoid profile, quantity and stability in 22 colour-fleshed potato cultivars grown in the Czech Republic. The total of nine carotenoids was analysed by HPLC using a C30 column and PDA detection. The total carotenoid content for all cultivars ranged from 1.44 to 40.13 μg/g DM. Yellow cultivars showed a much higher average total carotenoid content (26.22 μg/g DM) when compared to red/purple-fleshed potatoes (5.69 μg/g DM). Yellow cultivars were dominated by antheraxanthin, whereas neoxanthin was the main carotenoid in red/purple cultivars. Thermal processing significantly impacted all potato cultivars. Boiling decreased the total carotenoids by 92% compared to baking (88%). Lutein was the most stable carotenoid against thermal processing (decreased by 24-43%) followed by β-carotene (decreased by 78-83%); other carotenoids were degraded nearly completely. Increased formation of (Z)-isomers by thermal processing has not been confirmed.


Synthesis and Identification of Pregnenolone Derivatives as Inhibitors of Isozymes of 5α-Reductase.
Hyperplasia of the prostate gland and prostate cancer have been associated with high levels of serum 5α-dihydrotestosterone. This steroid is formed from testosterone by the activity of the enzyme 5α-reductase (5α-R) present in the prostate. Thus, inhibition of this enzyme could be a goal for therapies to treat these diseases. This study reports the synthesis and effects of five different 21-esters of pregnenolone derivatives as inhibitors of 5α-R types 1 and 2. The activity of these steroidal compounds was determined using in vivo and in vitro experiments. The results indicate that of the five steroids studied, the 21(p-fluoro)benzoyloxypregna-4,16-diene-3,6,20-trione derivative, whose structure has not yet been reported, has the best molecular conformation to inhibit the in vitro activity of both types of 5α-R. In addition, this steroid also displayed activity in vivo. Apparently, its pharmacological effect was increased by the presence of a keto group at C-6, because this group decreased the possibility that the steroid would be metabolized by hepatic enzymes. In addition, the double bond present at C-4 of this compound also enhanced its inhibitory activity on 5α-R, and the C-21 ester moiety increased its liphophilicity. Therefore, its solubility in the cell membrane and its pharmacological activity were both increased.


Hemodialysis-associated amyloidosis with cervical spinal cord compression and incomplete tetraplegia: a case report.
Case report. To report a rare case of hemodialysis-associated amyloidosis of cervical spinal canal with incomplete tetraplegia in long-term hemodialysis patient. Traumatology Clinic, Zagreb; Spinal Unit, Varazdinske Toplice, Croatia. Forty-seven-year-old male patient at long-term hemodialysis treatment developed progressive limbs weakness, graded as C4 ASIA C. Cervical computed tomography myelography showed extreme narrowing of the spinal canal. Decompressive laminectomy with bilateral foraminotomies was performed and histological examination revealed amyloidal deposition. Rehabilitation program started immediately after surgery with physical and occupational therapy, along with psychological support aiming at restoring full activities of daily living. Three months after surgery, the patient returned to his community, neurologically improved to ASIA E. Hemodialysis program was regularly maintained perioperatively and during the rehabilitation. Vertebral involvement with neurological deterioration in hemodialysis-associated amyloidosis patients prompts for early diagnosis, surgical and rehabilitation management, in this instance with good outcome.Spinal


Active bead-linked immunoassay on protein microarrays.
Protein microarrays are becoming a powerful tool in proteome, biochemical, and clinical studies. In addition to the quality of arrayed immobilized probe molecules, sensitivity of the microarray-based assay is highly dependent on the detection technique. Here we suggest four simple techniques for rapid detection of analytes bound to protein microarrays. The techniques employ functionalized magnetic and non-magnetic beads moved to, from, or along the array surface by external forces. In contrast to other labeling techniques actively controlled physical labels: (i) make detection extremely fast to allow microarray reading in seconds; (ii) provide a low background due to active removal of weakly bound beads; and (iii) provide a highly sensitive detection, since one antigen-antibody bond is capable of holding bead immobilized on the array surface. In combination with the electrophoretically assisted active immunoassay we described recently such active reading allows to reduce total indirect immunoassay time to 7-10 min while having sensitivity in the femtomolar concentration range. High speed, sensitivity, and specificity make active bead-linked detection an ideal choice in rapid high-throughput screening and in emergency diagnostics.


Neuroectodermal tumours in the cerebellum in two sisters.
Two sisters, one 5 years and the other 2 years old, with intrinsic tumours of the cerebellum are reported. One of the tumours was interpreted as being a ganglioneuroma with neuroblastomatous change. The other was a desmoplastic medulloblastoma.


Liver transplantation as a cure for acute intermittent porphyria.
Acute intermittent porphyria occasionally causes frequent and crippling acute neurovisceral attacks associated with increased hepatic production of porphyrin precursors, resulting in long-term damage, poor quality of life, and shortened life expectancy. There has been no cure for this condition, but replacement of deficient hepatic enzymes might restore excretion of porphyrin precursors to normal and prevent acute attacks. We aimed to treat severe acute intermittent porphyria in a 19-year-old woman by liver transplantation. After the transplant, concentrations of haem precursors in the patient's urine returned to normal, and 1.5 years later her quality of life was good. Our report suggests some hope of cure for selected patients with severe forms of this disease.


Induction of a spectroscopically defined transition by guanidinium hydrochloride on a recombinant calcium binding protein from Entamoeba histolytica.
Sequence analysis and metal ion binding studies reported earlier have established that the calcium binding protein (CaBP) from the parasitic ameboid Entamoeba histolytica protein has four canonical EF hand motifs which bind calcium. Equilibrium denaturation studies on both the apo and the holo forms of this protein indicate the presence of stable transition intermediates at low denaturant concentrations as revealed by the binding of the non-specific hydrophobic dye ANS. Fast reaction kinetics shows that the binding of the Gdn+ ions at or near the Ca2+ sites in the N-terminal domain influences metal ion binding to the sites in the C-terminal domain. Isothermal calorimetric titrations performed using low GdnHCl concentrations reveal the presence of two binding sites of low affinity, both being endothermic in nature. Thus the stabilization of CaBP observed at low GdnHCl concentration represents a native-like intermediate, with the Gdn+ ions mimicking Ca2+ binding at the N-terminal domain of this protein.


Exposure of Deinococcus radiodurans to both static magnetic fields and gamma radiation: observation of cell recuperation effects.
The extremophilic bacterium Deinococcus radiodurans displays an extraordinary ability to withstand lethal radiation effects, due to its complex mechanisms for both proteome radiation protection and DNA repair. Published results obtained recently at this laboratory show that D. radiodurans submitted to ionizing radiation results in its DNA being shattered into small fragments which, when exposed to a "static electric field' (SEF), greatly decreases cell viability. These findings motivated the performing of D. radiodurans exposed to gamma radiation, yet exposed to a different exogenous physical agent, "static magnetic fields" (SMF). Cells of D. radiodurans [strain D.r. GY 9613 (R1)] in the exponential phase were submitted to 60Co gamma radiation from a gamma cell. Samples were exposed to doses in the interval 0.5-12.5 kGy, while the control samples were kept next to the irradiation setup. Exposures to SMF were carried out with intensities of 0.08 T and 0.8 T delivered by two settings: (a) a device built up at this laboratory with niobium magnets, delivering 0.08 T, and (b) an electromagnet (Walker Scientific) generating static magnetic fields with intensities from 0.1 to 0.8 T. All samples were placed in a bacteriological incubator at 30 °C for 48 h, and after incubation, a counting of colony forming units was performed. Two sets of cell surviving data were measured, each in triplicate, obtained in independent experiments. A remarkable similarity between the two data sets is revealed, underscoring reproducibility within the 5% range. Appraisal of raw data shows that exposure of irradiated cells to SMF substantially increases their viability. Data interpretation strongly suggests that the increase of D. radiodurans cell viability is a sole magnetic physical effect, driven by a stochastic process, improving the efficiency of the rejoining of DNA fragments, thus increasing cell viability. A type of cut-off dose is identified at 10 kGy, above which the irradiated cellular system loses recovery and the cell survival mechanism collapses.


[Organ blood flow in graduated, controlled hypotension induced with sodium nitroprusside].
Metabolic acidosis is a characteristic feature of SNP-induced hypotension; this acidosis is of hepatic origin and is not due to underperfusion of the organ but to cyanide-induced anaerobic metabolism. For SNP doses higher than 3 mg/kg body weight, lactate is produced in the myocardium, brain, and skeletal muscles. The observed changes were reversible after hypotension of 20 min duration.


Case report: arthroscopic repair of a type IV SLAP lesion--the red-on-white lesion as a component of anterior instability.
This case report presents a rationale and technique for repair of a vascularized red-on-white bucket-handle tear of the glenoid labrum and intraarticular biceps tendon [type IV superior labrum anterior-to-posterior (SLAP) lesion]. This lesion may occur as a component of anterior instability, and as such it demands repair at the same time as anterior surgical stabilization for instability.


Impact of KRAS, BRAF and PI3KCA mutations in rectal carcinomas treated with neoadjuvant radiochemotherapy and surgery.
Conventional treatment for locally advanced rectal cancer usually combines neoadjuvant radiochemotherapy and surgery. Until recently, there have been limited predictive factors (clinical or biological) for rectal tumor response to conventional treatment. KRAS, BRAF and PIK3CA mutations are commonly found in colon cancers. In this study, we aimed to determine the mutation frequencies of KRAS, BRAF and PIK3CA and to establish whether such mutations may be used as prognostic and/or predictive factors in rectal cancer patients. We retrospectively reviewed the clinical and biological data of 98 consecutive operated patients between May 2006 and September 2009. We focused in patients who received surgery in our center after radiochemotherapy and in which tumor samples were available. In the 98 patients with a rectal cancer, the median follow-up time was 28.3 months (4-74). Eight out of ninety-eight patients experienced a local recurrence (8%) and 17/98 developed distant metastasis (17%). KRAS, BRAF and PIK3CA were identified respectively in 23 (23.5%), 2 (2%) and 4 (4%) patients. As described in previous studies, mutations in KRAS and BRAF were mutually exclusive. No patient with local recurrence exhibited KRAS or PIK3CA mutation and one harbored BRAF mutation (12.5%). Of the seventeen patients with distant metastasis (17%), 5 were presenting KRAS mutation (29%), one BRAF (5%) and one PIK3CA mutation (5%). No relationship was seen between PIK3CA, KRAS or BRAF mutation and local or distant recurrences. The frequencies of KRAS, BRAF and PIK3CA mutations in our study were lower than the average frequencies reported in colorectal cancers and no significant correlation was found between local/distant recurrences and KRAS, BRAF or PIK3CA mutations. Future studies with greater number of patients, longer follow-up time and greater power to predict associations are necessary to fully understand this relationship.


Association of CYBB polymorphisms with tuberculosis susceptibility in the Chinese Han population.
Reactive oxygen species (ROS) play a major role in the nonspecific innate immune response to invading microorganisms, such as Mycobacterium tuberculosis (MTB). Gp91phox, encoded by CYBB, serves as a key functional subunit of the Nicotinamide Adenine Dinucleotide Phosphate (NADPH) oxidase complex, which is pivotal to ROS generation. Therefore, the aim of the study was to investigate the association of CYBB polymorphisms with tuberculosis (TB) susceptibility. In total, 636 TB patients and 608 healthy, age and gender matched controls were enrolled in this study. All subjects were unrelated ethnic Han Chinese. Two tagSNPs were selected from the HapMap database and genotyped using matrix-assisted laser desorption/ionization time of flight mass spectrometry. After adjusting for confounders including age, gender and smoking, rs5917471 allele T showed significant association with decreased risk of TB (OR 0.745, 95% CI 0.556-0.999) and pulmonary TB (OR 0.618, 95% CI 0.410-0.931). However, no difference in allelic distribution was observed for the rs6610650 G/A polymorphism with respect to TB or different clinical types of TB. Further stratified analyses demonstrated the protective effect of allele T of rs5917471 was stronger among males (OR 0.500, 95% CI 0.295-0.846), smokers (OR 0.462, 95% CI 0.239-0.896), and male smokers (OR 0.372, 95% CI 0.182-0.761); the individuals carrying the A allele of rs6610650 exhibited an decreased risk of TB among males, smokers and male smokers, with OR (95% CI) of 0.535 (0.290-0.984), 0.442 (0.198-0.988), and 0.350 (0.145-0.845), respectively. There were no statistically significant differences in haplotype distribution between TB and control groups. Smoking and rs5917471 formed the best gene-environment interaction model with the testing balanced accuracy of 53.29% and cross-validation consistency of 9/10. This is the first study of the association of CYBB polymorphisms with TB. Our findings suggest that the CYBB polymorphisms are significantly correlated with reduced risk of TB, especially among male smokers. Further studies are needed to verify this association.


Unexpected structures formed by the kinase RET C634R mutant extracellular domain suggest potential oncogenic mechanisms in MEN2A.
The RET receptor tyrosine kinase plays a pivotal role in cell survival, proliferation, and differentiation, and its abnormal activation leads to cancers through receptor fusions or point mutations. Mutations that disrupt the disulfide network in the extracellular domain (ECD) of RET drive multiple endocrine neoplasia type 2A (MEN2A), a hereditary syndrome associated with the development of thyroid cancers. However, structural details of how specific mutations affect RET are unclear. Here, we present the first structural insights into the ECD of the RET(C634R) mutant, the most common mutation in MEN2A. Using electron microscopy, we demonstrate that the C634R mutation causes ligand-independent dimerization of the RET ECD, revealing an unusual tail-to-tail conformation that is distinct from the ligand-induced signaling dimer of WT RET. Additionally, we show that the RETC634R ECD dimer can form complexes with at least two of the canonical RET ligands and that these complexes form very different structures than WT RET ECD upon ligand binding. In conclusion, this structural analysis of cysteine-mutant RET ECD suggests a potential key mechanism of cancer induction in MEN2A, both in the absence and presence of its native ligands, and may offer new targets for therapeutic intervention.


Simultaneous LC determination of paracetamol and related compounds in pharmaceutical formulations using a carbon-based column.
A simple, rapid and convenient high performance liquid chromatographic method, which permits the simultaneous determination of paracetamol, 4-aminophenol and 4-chloracetanilide in pharmaceutical preparation has been developed. The chromatographic separation was achieved on porous graphitized carbon (PGC) column using an isocratic mixture of 80/20 (v/v) acetonitrile/0.05 M potassium phosphate buffer (pH 5.5) and ultraviolet detection at 244 nm. Correlation coefficient for calibration curves in the ranges 1-50 microg ml(-1) for paracetamol and 5-40 microg ml(-1) for 4-aminophenol and 4-chloroacetanilide were >0.99. The sensitivity of detection is 0.1 microg ml(-1) for paracetamol and 0.5 microg ml(-1) for 4-aminophenol and 4-chloroacetanilide. The proposed liquid chromatographic method was successfully applied to the analysis of commercially available paracetamol dosage forms with recoveries of 98-103%. It is suggested that the proposed method should be used for routine quality control and dosage form assay of paracetamol in pharmaceutical preparations. The chromatographic behaviour of the three compounds was examined under variable mobile phase compositions and pH, the results revealed that selectivity was dependent on the organic solvent and pH used. The retention selectivity of these compounds on PGC was compared with those of octadecylsilica (ODS) packing materials in reversed phase liquid chromatography. The ODS column gave little separation for the degradation product (4-aminophenol) from paracetamol, whereas PGC column provides better separation in much shorter time.


Safety and Immunogenicity of M-M-RII (Combination Measles-Mumps-Rubella Vaccine) in Clinical Trials of Healthy Children Conducted Between 1988 and 2009.
M-M-RII, a combination measles, mumps and rubella vaccine, was licensed in the United States in 1978 based on data from several clinical trials that demonstrated that the safety and immunogenicity of the vaccine were comparable to the component monovalent vaccines and to the previous trivalent combination vaccine. Safety and immunogenicity data from 23 postlicensure clinical trials conducted with M-M-RII between 1988 and 2009 were summarized. A total of 12,901 children who received only a first dose, 920 children who received a first and second dose and 400 children who received only a second dose were evaluated. The vaccine was generally well tolerated among children who received a first and/or second dose of M-M-RII. During the 28-42-day follow-up after dose 1 and dose 2, the median rate of temperatures ≥102°F (oral equivalent) was 24.8% and 13.0% and the median rate of measles/rubella-like rash was 3.2% and 0.5%, respectively. The median rate of injection-site reactions during the first 5 days postdose 1 and postdose 2 was 17.3% and 42.7%, respectively. The seroconversion rates (enzyme-linked immunosorbent assay) after dose 1 were remarkably consistent from study to study between 1988 and 2009 (92.8%-100% for measles, 97.7%-100% for mumps and 92.8%-100% for rubella). A trend test showed that there was no change in the immunogenicity of the vaccine over the 21-year period. The results of this analysis demonstrate that M-M-RII is well tolerated and immunogenic. The vaccine performed consistently over 21 years of evaluation in clinical trials.


Long term follow-up of complement parameters to improve the management of acquired angioedema due to C1-inhibitor deficiency.
Acquired angioedema due to C1-inhibitor deficiency (C1-INH-AAE) is a rare disease that can be diagnosed via complement testing. It often accompanies lymphoproliferative underlying diseases. Our study aimed to examine if there is a connection between complement parameters and the clinical symptoms of C1-INH-AAE, and, in case of a known underlying disease, its activity. The other question is how a connection, if proven, could help in the development of the therapeutic strategy of C1-INH-AAE patients. In the past 30 years, out of the 3938 patients sent to the Angioedema Center with angioedema symptoms, we have diagnosed C1-INH-AAE in 19 cases. An underlying disease was diagnosed in 15 patients. Most often lymphoma (6/19 patients) and monoclonal gammopathy of undetermined significance (6/19 patients) were found. Angioedema specific long-term prophylaxis did not result in an improvement in neither the frequency of the attacks nor in the complement parameters. A connection has been found between the presence and activity of any underlying disease, the frequency of the angioedema attacks and the decreased level of proteins of the complement system. Decreasing complement parameters warn about the appearance or the worsening of the underlying disease. The treatment of the underlying disease brings improvement in the complement parameters. Rituximab treatment reduced the number of attacks or completely made them disappear, and we experienced positive changes in complement parameters. Complement parameters supported the long-term efficacy of rituximab treatment for C1-INH-AAE. The change in complement parameters predict the relapse of the underlying disease, and it is a good indicator for the prediction of angioedematous attacks. In C1-INH-AAE, it is essential to examine the patients for underlying diseases, and to regularly follow up the patient's complement parameters.


Doxorubicin and beta-lapachone release and interaction with micellar core materials: experiment and modeling.
Polymer micelles with two different core-forming blocks, poly(d,l -lactide) (PLA) and poly(epsilon-caprolactone) (PCL), but the same coronal material, poly(ethylene glycol) (PEG), were investigated in this study as nanoscopic drug carriers. The release of two different drugs, doxorubicin (DOX) and beta-lapachone (beta-lap), from PEG(5k)-b-PCL(5k) and PEG(5k)-b-PLA(5k) micelles was studied at pH 5.0 and 7.4. Mathematical solutions of both Higuchi's model and Fickian diffusion equations were utilized to elucidate the differences between the micelle core materials for the two drugs. The neutral and smaller of the two drugs tested, beta-lap, demonstrated faster, pH-independent release, suggesting that no substantial changes occurred in either micelle core at lower pH. In contrast, the release rate of DOX was found to noticeably increase at lower pH with a larger cumulative amount of drug released. Different core materials were shown to have considerable influence on the release kinetics of both drugs: in both cases, the more hydrophobic PCL core showed slower drug release rates compared with the less hydrophobic PLA core.


Abuse liability of flunitrazepam among methadone-maintained patients.
Abuse liability and acute subjective and psychomotor effects of flunitrazepam were assessed in ten methadone-maintained males with history of benzodiazepine and alcohol use, who voluntarily participated in a double-blind, controlled, cross-over, randomized clinical trial. There were six experimental sessions in which a single oral dose of flunitrazepam 1, 2, and 4 mg; triazolam 0.5 and 0.75 mg; and placebo was given. Evaluations included physiological measures; psychomotor performance tasks (simple reaction time, Digit Symbol Substitution Test, balance task, Maddox-wing device); and self-administered subjective effects questionnaires [Addiction Research Center Inventory (ARCI), Profile of Mood States (POMS), a series of visual analog scales (VAS)]. All drugs but flunitrazepam 1 mg caused an impairment of psychomotor tasks. Effects were more evident with the highest doses of both drugs. Only flunitrazepam 4 mg produced a significant decrease in balance time. Triazolam 0.75 mg induced increases in sedation measured by ARCI-PCAG, depression in POMS, and VAS-drowsiness scores. Flunitrazepam 4mg caused euphoria-related effects as measured by increases in ARCI-MBG and "high" scores in the VAS. Our findings of flunitrazepam-induced euphoria in methadone-maintained subjects together with epidemiological evidence of flunitrazepam abuse by opioid dependents, suggest that it may be included in the group of benzodiazepines with a relatively high abuse potential.


[Potentialities of an x-ray method of determining malignant changes in polyps of the large intestine].
The study was concerned with evaluation of the clinical and roentgenologic data on 57 cases of malignant polyps of the colon. Malignant polyps reaching 10 mm in length were found. Malignancies were identified using endoscopy and biopsy in 58.6% and X-ray examination--45.8%. Among the most effective methods of X-ray examination of the large bowel for cancer should be: high-voltage roentgenography employing incomplete barium enema, primary double contrast examination, direct blow-up and parietography. X-ray signs of malignant transformation of polyps are: persistent retention of barium, wavy and indistinct contours, irregular pattern, rapid growth of neoplasm, retraction of tumor base and disparity between polyp size and its pedicle length.


Development of the heartbeat during normal ontogeny and during long-term organ culture of hearts of the newt, Cynops pyrrhogaster.
Development of cardiac contraction was investigated in the newt, Cynops pyrrhogaster, during normal development and in hearts maintained in long-term organ culture. In the embryos the heart began beating as early as stage 33. The heart rate increased up to stage 52 and then gradually decreased to the adult rate by stage 60. The heart rate showed a logarithmic temperature dependence. In organ-cultured hearts the pattern of heart rate depended on the stage at which the heart was explanted; cultured embryonic hearts showed a pattern of increase and decline similar to that seen in vivo, and hearts explanted near metamorphosis showed only a slow decline.


Oral Delivery of Nanoparticles Loaded With Ginger Active Compound, 6-Shogaol, Attenuates Ulcerative Colitis and Promotes Wound Healing in a Murine Model of Ulcerative Colitis.
Oral drug delivery is the most attractive pathway for ulcerative colitis [UC] therapy, since it has many advantages. However, this strategy has encountered many challenges, including the instability of drugs in the gastrointestinal tract [GT], low targeting of disease tissues, and severe adverse effects. Nanoparticles capable of colitis tissue-targeted delivery and site-specific drug release may offer a unique and therapeutically effective system that addresses these formidable challenges. We used a versatile single-step surface-functionalising technique to prepare PLGA/PLA-PEG-FA nanoparticles loaded with the ginger active compound, 6-shogaol [NPs-PEG-FA/6-shogaol]. The therapeutic efficacy of NPs-PEG-FA/6-shogaol was evaluated in the well-established mouse model of dextran sulphate sodium [DSS]-induced colitis. NPs-PEG-FA exhibited very good biocompatibility both in vitro and in vivo. Subsequent cellular uptake experiments demonstrated that NPs-PEG-FA could undergo efficient receptor-mediated uptake by colon-26 cells and activated Raw 264.7 macrophage cells. In vivo, oral administration of NPs-PEG-FA/6-shogaol encapsulated in a hydrogel system [chitosan/alginate] significantly alleviated colitis symptoms and accelerated colitis wound repair in DSS-treated mice by regulating the expression levels of pro-inflammatory [TNF-α, IL-6, IL-1β, and iNOS] and anti-inflammatory [Nrf-2 and HO-1] factors. Our study demonstrates a convenient, orally administered 6-shogaol drug delivery system that effectively targets colitis tissue, alleviates colitis symptoms, and accelerates colitis wound repair. This system may represent a promising therapeutic approach for treating inflammatory bowel disease [IBD].


[Use of dehydrated temporal fascia in chronic otitis media surgery and tympanoplasty].
We evaluated the use of homograft dehydrated temporal fascia (Tutoplast) in repair of tympanic membrane perforations secondary to chronic otitis media. Dehydrated temporal fascia was used during tympanoplasty in 17 patients (10 females, 7 males; mean age 34 years; range 15 to 53 years) with chronic otitis media. Operations were type 1 tympanoplasty in six patients, tympanoplasty and mastoidectomy in three patients, and revision tympanoplasty and/or mastoidectomy in the remaining patients. The patients were followed-up for at least two years. Complete wound healing was achieved without complications, and the material used was well tolerated in all the patients. Only one patient had a small perforation. The tympanic membranes were anatomically intact in 16 patients (94%). The use of dehydrated temporal fascia provides maximum microsurgical comfort due to its soft, flexible feature and maintenance of its delicacy even in contact with blood. Thus, dehydrated temporal fascia should be considered for selected cases for its ease of use and anatomical success.


Brachial plexus dose tolerance in head and neck cancer patients treated with sequential intensity modulated radiation therapy.
We aimed to study the radiation induced brachial plexopathy in patients with head and neck squamous cell carcinoma (HNSCC) treated with Sequential Intensity Modulated Radiation Therapy (S-IMRT). This IRB approved study included 68 patients with HNSCC treated consecutively. Detailed dose volume histogram data was generated for ipsilateral and contralateral brachial plexus (BP) volumes receiving a specified dose (Vds) i.e. V50-V75 and dose in Gray covering specified percent of BP volume (Dvs) i.e. D5-D30 and maximum point doses (Dmax). To assess BP injury all patients' charts were reviewed in detail for sign and symptoms of BP damage. Post-hoc comparisons were done using Tukey-Kramer method to account for multiple significance testing. The mean and maximum doses to BP were significantly different (p < .05) based on tumor site, nodal status and tumor stage. The mean volume to the ipsilateral BP for V50, V60, V70, and V75 were 7.01 cc, 4.37 cc, 1.47 cc and 0.24 cc, respectively. The mean dose delivered to ≤5% of ipsilateral BP was 68.70 Gy (median 69.5Gy). None of the patients had acute or late brachial plexopathy or any other significant neurological complications, with a minimum follow up of two years (mean 54 months). In this study cohort, at a minimum of two-years follow up, the mean dose of 68.7Gy, a median dose to 69.5Gy to ≤5% of ipsilateral BP, and a median Dmax of 72.96Gy did not result in BP injury when patients were treated with S-IMRT technique. However, longer follow up is needed.


Stability and predictive value of anti-JCV antibody index in multiple sclerosis: A 6-year longitudinal study.
Risk of natalizumab-related progressive multifocal leukoencephalopathy is associated with the presence of anti-JC-virus (JCV) antibodies. To investigate the longitudinal evolution of anti-JCV antibody index and to determine the predictive value of baseline anti-JCV antibody index for long-term stability of anti-JCV antibody status. MS patients from the MS centre of Medical University of Innsbruck, who had serum sampling for a time period of 4-6 years at intervals of 6±3 months, were included in this retrospective, longitudinal study. Anti-JCV antibody serological status and index were determined by 2-step second-generation anti-JCV antibody assay. 154 patients were included in this study. Median follow-up time was 63.7 months, with median 11 samples available per patient. At baseline, 111 (72.1%) patients were anti-JCV antibody positive. Baseline anti-JCV antibody index significantly correlated with age (R = 0.22, p = 0.005); there was no difference with respect to sex, disease duration or previously used disease-modifying treatment. During follow-up anti-JCV antibody status changed from negative to positive or vice versa in 17% of patients. In seronegative patients at baseline, baseline anti-JCV antibody index was significantly lower in those remaining seronegative at follow-up compared to those converting to seropositivity (median 0.16 vs. 0.24, p = 0.002). In seropositive patients at baseline, index was higher in those remaining seropositive compared to those reverting to seronegativity (2.6 vs. 0.45, p<10-7). Baseline anti-JCV antibody index >0.90 predicted stable positive serostatus (sensitivity 88.7%, specificity 96.5%) and <0.20 stable negative serostatus (sensitivity 61.3%, specificity 97.6%). Anti-JCV antibody index remained relatively stable over 6-year follow-up with annual serostatus change of ~3%. Baseline anti-JCV antibody index predicted stable negative and stable positive JCV serostatus.


Preparing a Workforce of Care Coordinators to Address Patient Mental Health Needs in the Digital Age: Training and Needs Identification.
Care coordinators (CCs) are specialized healthcare providers and often the primary point of contact for patients with multiple medical and mental health comorbidities in integrated healthcare settings. Prior work shows CCs have lower comfort addressing mental health than physical health concerns. Digital mental health interventions can support CCs' management of patient mental health needs, but training gaps must be addressed prior to a digital mental health intervention's implementation. As part of a quality improvement initiative, a 1-hour training focused on the assessment and management of depression and suicide-related thoughts and behaviors was delivered to CCs within a large midwestern healthcare system's Division of Ambulatory Care Coordination. CCs completed online surveys prior to and following the training. Training resulted in increased comfort working with clinical populations, including patients who experience suicide-related thoughts and behaviors. Gains around screening for suicide risk were modest. Brief trainings for CCs can address training gaps, however, ongoing training and case consultation may also be indicated.


Diarrhea in tube-fed patients: feeding formula not necessarily the cause.
This study of diarrhea in tube-fed patients was undertaken to determine the proportion of cases in which feeding formula is not responsible for the diarrhea, the causes other than the feeding formula, and the diagnostic approach to diarrhea in tube-fed patients. Inpatients at the Truman Memorial Veterans Hospital who received nasoenteric feeding during the time period from October 1986 through May 1988 were eligible for this study. Of 123 patients who received nasoenteric feeding, 32 patients had documented diarrhea (greater than 500 mL per day for at least two consecutive days) and were enrolled. Three of these patients received hypertonic feeding formula, whereas the remaining 29 received isotonic feeding formula. Prospective determinations of the causes of diarrhea were performed. Laboratory tests included fecal leukocytes, stool osmolality, stool electrolytes, and Clostridium difficile toxin assay. Diarrhea was considered osmotic if the stool osmotic gap was greater than 100 mmol/L. Clinical management involved reducing or stopping the feeding formula, stopping suspected medications, or administering appropriate antibiotics. There were 32 episodes of diarrhea in tube-fed patients during the study period. A single cause could be specified in 29 cases. The tube feeding formula was responsible for diarrhea in only 21% of these cases. Medications were directly responsible in 61% and C. difficile in 17% of cases. Stool osmotic gap correctly distinguished osmotic from non-osmotic diarrhea in all cases. When diarrhea develops in properly tube-fed patients, the feeding formula is usually not responsible for the diarrhea. Patients receiving nasoenteric tube feeding are frequently placed on liquid forms of medications. Many medicinal elixirs contain sorbitol, which is often the cause of diarrhea in tube-fed patients. Review of the medications and determination of the stool osmotic gap are the initial diagnostic steps of highest yield.


Automated turbidimetry for rapid determination of the bacteriological quality of raw meat and processed meat products.
A fully automated centrifugal spectrophotometer for turbidimetric measurements linked to a computer for data processing was used to record and analyze growth of different types of bacteria associated with spoilage of raw meat and processed meat products (sliced meat for sandwiches) in order to predict bacterial counts. Media for this specific analysis were developed to enumerate total viable count, lactic acid bacteria. Brochothrix thermosphacta and Pseudomonas spp. Correlation coefficients of -0.91 to -0.97 were obtained between the conventional plate count technique and the turbidimetric method (linear regression). Lactic acid bacteria in the range 10(3) to 10(9) cfu/g could be detected within 48 h, whereas total viable count, Brochothrix thermosphacta and Pseudomonas spp. generally could be detected within 24 h. Counts exceeding 10(7) cfu/g could be detected within 6 h.


Faculty Development for Fostering Clinical Reasoning Skills in Early Medical Students Using a Modified Bayesian Approach.
Clinical reasoning is a necessary skill for medical students to acquire in the course of their education, and there is evidence that they can start this process at the undergraduate level. However, physician educators who are experts in their given fields may have difficulty conveying their complex thought processes to students. Providing faculty development that equips educators with tools to teach clinical reasoning may support skill development in early medical students. We provided faculty development on a modified Bayesian method of teaching clinical reasoning to clinician educators who facilitated small-group, case-based workshops with 2nd-year medical students. We interviewed them before and after the module regarding their perceptions on teaching clinical reasoning. We solicited feedback from the students about the effectiveness of the method in developing their clinical reasoning skills. We carried out this project during an institutional curriculum rebuild where clinical reasoning was a defined goal. At the time of the intervention, there was also increased involvement of the Teaching and Learning Center in elevating the status of teaching and learning. There was high overall satisfaction with the faculty development program. Both the faculty and the students described the modified Bayesian approach as effective in fostering the development of clinical reasoning skills. Through this work, we learned how to form a beneficial partnership between a clinician educator and Teaching and Learning Center to promote faculty development on a clinical reasoning teaching method for early medical students. We uncovered challenges faced by both faculty and early learners in this study. We observed that our faculty chose to utilize the method of teaching clinical reasoning in a variety of manners in the classroom. Despite obstacles and differing approaches utilized, we believe that this model can be emulated at other institutions to foster the development of clinical reasoning skills in preclerkship students.


NEU3 sialidase strictly modulates GM3 levels in skeletal myoblasts C2C12 thus favoring their differentiation and protecting them from apoptosis.
Membrane-bound sialidase NEU3, often referred to as the "ganglioside sialidase," has a critical regulatory function on the sialoglycosphingolipid pattern of the cell membrane, with an anti-apoptotic function, especially in cancer cells. Although other sialidases have been shown to be involved in skeletal muscle differentiation, the role of NEU3 had yet to be disclosed. Herein we report that NEU3 plays a key role in skeletal muscle differentiation by strictly modulating the ganglioside content of adjacent cells, with special regard to GM3. Induced down-regulation of NEU3 in murine C2C12 myoblasts, even when partial, totally inhibits their capability to differentiate by increasing the GM3 level above a critical point, which causes epidermal growth factor receptor inhibition (and ultimately its down-regulation) and an higher responsiveness of myoblasts to the apoptotic stimuli.