Residents in trouble: an in-depth assessment of the 25-year experience of a single family medicine residency.
Graduate medical education program directors struggle with resident trainees who develop problems in academic and personal spheres leading to probation, resignation, or termination of training. These "residents in trouble" generate a disproportionate amount of work and can demoralize faculty and residents. Only two prior studies have identified the prevalence of residents in trouble. In both studies, the prevalence was based on nonvalidated program director self-report. In contrast, our study uses a review of primary data sources to describe the prevalence, types of problems, remediation methods, and outcomes for residents in trouble over the 25-year experience of a residency program. Five primary data sources were reviewed for every trainee who was enrolled from 1978-2003 at the David Grant Medical Center Family Medicine Residency, looking for any of eight types of occurrences previously identified in the literature as marking a resident in trouble. Over this 25-year period, 230 residents entered training and 209 graduated. The prevalence of residents in trouble was 9.1%. The types of problems encountered included divorce, psychiatric illness, substance abuse, attitudinal problems, interpersonal conflict, excessive patient complaints, and insufficient knowledge. A variety of remediation approaches were used, and 90% of the residents in trouble graduated. The 9.1% provides a point of comparison for program directors to know if their residency has a disproportionate rate of problems requiring organizational reform or if they are dealing with the normative experience of a training program. Our program's experience supports five suggestions for the management of residents in trouble.


[Role of alpha-interferon in regulation of apoptosis in cells of the hypothalamo-hypophyseal-adrenocortical system of old mice in oxidative stress].
The aim of this work was to demonstrate the role of interferon alpha (IA) in apoptosis control of cells of hypothalamohypophysial adrenocortical system (HHAS) in young and old mice under conditions of hyperoxia. The increase of functional activity of neurosecretory cells of hypothalamic paraventricular nucleus in old mice treated with IA was demonstrated, while no effect was found in young mice. This differences could be associated with aging changes in HHAS in intact old animals. Oxidative stress caused an increase in the number of cells synthesizing proapoptotic c-fos protein in paraventricular nucleus of mice of both age groups. However, the protective action of IA against the stress is expressed more actively and in more early stage of apoptosis in young mice. Thus, the degree of IA participation in the control of programmed cell death of hypothalamic cells depends on the age of the animal. In the adrenal zona fasciculata of young mice the number of apoptotic cells was significantly increased after IA administration while after hyperoxia or its combination with IA it remained at control level. In the adrenal gland of old mice the proportion of apoptotic cells was not different from that found in young mice and remained unchanged after all experimental challenges.


Tight coupling of Na+/K+-ATPase with glycolysis demonstrated in permeabilized rat cardiomyocytes.
The effective integrated organization of processes in cardiac cells is achieved, in part, by the functional compartmentation of energy transfer processes. Earlier, using permeabilized cardiomyocytes, we demonstrated the existence of tight coupling between some of cardiomyocyte ATPases and glycolysis in rat. In this work, we studied contribution of two membrane ATPases and whether they are coupled to glycolysis--sarcoplasmic reticulum Ca2+ ATPase (SERCA) and plasmalemma Na+/K+-ATPase (NKA). While SERCA activity was minor in this preparation in the absence of calcium, major role of NKA was revealed accounting to ∼30% of the total ATPase activity which demonstrates that permeabilized cell preparation can be used to study this pump. To elucidate the contribution of NKA in the pool of ATPases, a series of kinetic measurements was performed in cells where NKA had been inhibited by 2 mM ouabain. In these cells, we recorded: ADP- and ATP-kinetics of respiration, competition for ADP between mitochondria and pyruvate kinase (PK), ADP-kinetics of endogenous PK, and ATP-kinetics of total ATPases. The experimental data was analyzed using a series of mathematical models with varying compartmentation levels. The results show that NKA is tightly coupled to glycolysis with undetectable flux of ATP between mitochondria and NKA. Such tight coupling of NKA to PK is in line with its increased importance in the pathological states of the heart when the substrate preference shifts to glucose.


Developing hospital resilience domains in facing disruption era in Indonesia: a qualitative study.
The studies of hospital resilience have been of increasing importance during the last decade due to disasters and pandemics. However, studies in developing the domain and indicators of hospital resilience were limited mainly on disaster response. A few studies of hospital resilience focused on how to deal with disruptions such as environmental turbulence, rapid technological changes, and changes in patient preferences. This study aims to develop domains and indicators of hospital resilience in facing the disruption era. This qualitative study focused on exploring the domains and indicators to face disruptions that have been identified in the first exploratory phase of the studies. Key informants included hospital experts from the government, medical practitioners, and academics. A total of 20 key informants were involved in semi-structured interviews which were conducted face-to-face, via telephone and Zoom. Data was analyzed using a grounded theory approach to discover domains for a resilient hospital. The study identified a number of domains that are fundamental for a hospital to become a resilient in the face of disruption. These include readiness to face digital transformation, effective leadership, and flexibility in managing resources among others. Situation awareness and resilience ethos, implementation of marketing management, networking, and disaster anticipation are found to be equally important. These domains focused on the hospital's ability to deal with specific shocks from different perspectives as the result of changes from disruptions which are inevitable within the organizational business environment. The domains identified in the study are able to respond to the limitations of the concept of hospital resilience, which is currently more focused on hospital disaster resilience. They can be used to measure hospital resilience in the context of the volatility, uncertainty, complexity, and ambiguity (VUCA), which are relevant to the context of the Indonesia hospital industry.


Pediatric viral gastroenteritis during eight years of study.
During the period January 1974 through July 1982, fecal samples from 1,537 pediatric inpatients with gastroenteritis were tested for enteric viruses by electron microscopic and rotavirus enzyme-linked immunosorbent assay techniques. Rotaviruses were detected in 34.5% of these patients, enteric adenoviruses were detected in 4.7%, approximately 27-nm viruses were detected in 1.6%, and at least one of these agents was found in 40.1% of the study subjects. Three infections were by an apparently new agent which morphologically is a rotavirus, but which failed to react in the rotavirus enzyme-linked immunosorbent assay. During the first 8 calendar years of study, rotaviruses were detected in 39.0% of 577 patients in the even-numbered years and 30.3% of 702 patients in the odd-numbered years. Adenoviruses were found in all calendar months. Rotaviruses were found in inpatients in November through July, whereas approximately 27-nm viruses were found in October through June. The percentage of patients who had a demonstrated viral infection rose steadily from 7.4% in September to 72.0% in January and then steadily declined to 2.9% in August. Viral infection was especially common in study subjects who were 7 through 24 months of age; 61% of such children had one or more enteric viruses. Rotavirus-infected patients tended to be younger during the months of greatest rotavirus activity than at the beginning and end of the rotavirus season, presumably because of a greater exposure to virus at the height of the rotavirus outbreak.


1H-n.m.r. investigation of the interaction between cytochrome c and cytochrome b5.
The interaction between eukaryotic cytochrome c and the tryptic fragment of bovine liver microsomal cytochrome b5 was studied by 1H-n.m.r. spectroscopy, and a procedure was developed that may be generally applicable to the study of macromolecular interactions by n.m.r. At pH6.3 (27 degrees C, I approx. 0.04) the two ferricytochromes were found to form a 1:1 complex with an association constant of approx. 10(3) M -1. The protein--protein-interaction region was found to encompass the region of the surface of horse cytochrome c that includes Ile-81, Phe-82, Ala-83 and Ile-85, and Lys-13 and Lys-72 of horse cytochrome c were suggested to be involved in two important intermolecular interactions. Me3Lys-72 of Candida krusei cytochrome c was shown to be involved in the interaction.


Evidence linking microRNA suppression of essential prosurvival genes with hippocampal cell death after traumatic brain injury.
The underlying molecular mechanisms of how dysregulated microRNAs (miRNAs) cause neurodegeneration after traumatic brain injury (TBI) remain elusive. Here we analyzed the biological roles of approximately 600 genes - we previously found these dysregulated in dying and surviving rat hippocampal neurons - that are targeted by ten TBI-altered miRNAs. Bioinformatic analysis suggests that neurodegeneration results from a global miRNA-mediated suppression of genes essential for maintaining proteostasis; many are hub genes - involved in RNA processing, cytoskeletal metabolism, intracellular trafficking, cell cycle progression, repair/maintenance, bioenergetics and cell-cell signaling - whose disrupted expression is linked to human disease. Notably, dysregulation of these essential genes would significantly impair synaptic function and functional brain connectivity. In surviving neurons, upregulated miRNA target genes are co-regulated members of prosurvival pathways associated with cellular regeneration, neural plasticity, and development. This study captures the diversity of miRNA-regulated genes that may be essential for cell repair and survival responses after TBI.


[Morphological characteristics of the shell of the ova of Capillariidae (Nematoda, Capillariidae)].
Peculiarities of the surface architectonics of the egg shell in 18 species of capillariids (Nematoda, Capillariidae) were studied. Capillariids were found in mammals of 5 orders: Insectivora, Chiroptera, Muridae, Carnivora and Arctiodactyla. A correlation between the structure of the external surface of egg shell of these nematodes on one hand, and localization in the organism of the definitive host and its biology on the other, was established. On the basis of these characters capillariids of the genera Capillaria, Eucoleus, Calodium, Liniscus, Thominx and Scrjabinocapillaria were arranged into 6 groups: 1 - the genus Capillaria (luman of the stomach and gut), 2 - the genus Eucoleus (mucous membrane of oesophagus and stomach), 3 - the genus Calodium (parenchima of the liver), 4 - the genus Liniscus (urinary bladder), 5 - the genus Thominx (lungs), 6 - the genera Thominx and Scrjabinocapillaria (lumen of the stomach). The classification suggested makes more precise and supplements the known systems of nematodes of the fam. Capillariidae.


Clinical features of insulinoma in patients with multiple endocrine neoplasia type 1: analysis of the database of the MEN Consortium of Japan.
More than 50% of patients with multiple endocrine neoplasia type 1 (MEN1) develop gastroenteropancreatic neuroendocrine tumors (GEPNETs), and insulinoma is the second most common functioning GEPNET. Compared to other functioning and nonfunctioning GEPNETs in MEN1, insulinoma is considered to develop at a younger age. To clarify the clinical features of insulinoma developed in Japanese patients with MEN1, a recently constructed database of Japanese MEN1 patients was analyzed. Among 560 registered cases, insulinoma was seen in 69 patients and information on age at diagnosis was available for 54 patients. Tumors predominantly occurred in the body and tail of the pancreas. The mean age at diagnosis of insulinoma (34.8 ± 16.7 yrs) was significantly younger than that of gastrinoma (50.6 ± 14.3 yrs) and nonfunctioning tumor (44.7 ± 13.3 yrs) in patients with MEN1. Patients diagnosed as having insulinoma during middle-age (30 - 49 yrs) tended to have a long period from appearance of hypoglycemic symptoms to diagnosis of the tumor. Of note, 13 patients (24%) were diagnosed as having insulinoma before 20 yrs of age. Such young onset was not seen in other GEPNETs. Since the development of GEPNETs during adolescence is quite rare, insulinoma diagnosed before 20 yrs strongly suggests the presence of MEN1 and warrants further investigation, including MEN1 genetic testing. Also, clinicians should be aware that insulinoma can often be missed in middle-aged patients.


[Primary leiomyosarcoma of the lung--a case report].
One case of primary leiomyosarcoma of the lung is presented. Leiomyosarcoma of the lung is a rare tumor. Of 83 cases reported in the literatures, 54 were recorded in detail. The incidence of the disease in the male is higher than that in the female. Children, adolescents and young adults may be involved. Its clinical and X-ray manifestations are similar to those of lung cancer. The diagnosis is established by pathologic examination. Direct visualization, biopsy and brushing during bronchoscopy are important methods for preoperative diagnosis. The chance of positive sputum cytology is very small. The pathologic features of this disease are emphasized. It is suggested that surgery be more effective for this tumor than the other lung cancers.


Institutional analysis of health system governance.
It is important that researchers who study health system governance have a set of collective understandings of the meanings of governance, which can then inform the methods used in research. We present an institutional framing and definition of health system governance; that is, governance refers to making, changing, monitoring and enforcing the rules that govern the demand and supply of health services. This pervasive, relational view of governance is to be preferred to approaches that focus primarily on structures of governments and health care organizations, because health system governance involves communities and service users, and because governments in many low- and middle-income countries tend to under-govern. Therefore, the study of health system governance requires institutional analysis; an approach that focuses not only on structures, but also on the rules (both formal and informal) governing demand and supply relations. Using this 'structure-relations' lens, and based on our field experience, we discuss how this focus could be applied to the three approaches to framing and studying health system governance that we identified in the literature. In order of decreasing focus on structures ('hardware') and increasing focus on relations ('software'), they are: (1) the government-centred approach, which focuses on the role of governments, above or to the exclusion of non-government health system actors; (2) the building-block approach, which focuses on the internal workings of health care organizations, and treats governance as one of the several building blocks of organizations; and (3) the institutional approach, which focuses on how the rules governing social and economic interactions are made, changed, monitored and enforced. Notably, either or both qualitative and quantitative methods may be used by researchers in efforts to incorporate the analysis of how rules determine relations among health system actors into these three approaches to health system governance.


A multiple-scenario assessment of the effect of a continuous-care, guideline-based decision support system on clinicians' compliance to clinical guidelines.
To quantify the effect of a new continuous-care guideline (GL)-application engine, the Picard decision support system (DSS) engine, on the correctness and completeness of clinicians' decisions relative to an established clinical GL, and to assess the clinicians' attitudes towards a specific DSS. Thirty-six clinicians, including residents at different training levels and board-certified specialists at an academic OB/GYN department that handles around 15,000 deliveries annually, agreed to evaluate our continuous-care guideline-based DSS and to perform a cross-over assessment of the effects of using our guideline-based DSS. We generated electronic patient records that realistically simulated the longitudinal course of six different clinical scenarios of the preeclampsia/eclampsia/toxemia (PET) GL, encompassing 60 different decision points in total. Each clinician managed three scenarios manually without the Picard DSS engine (Non-DSS mode) and three scenarios when assisted by the Picard DSS engine (DSS mode). The main measures in both modes were correctness and completeness of actions relative to the PET GL. Correctness was further decomposed into necessary and redundant actions, relative to the guideline and the actual patient data. At the end of the assessment, a questionnaire was administered to the clinicians to assess their perceptions regarding use of the DSS. With respect to completeness, the clinicians applied approximately 41% of the GL's recommended actions in the non-DSS mode. Completeness increased to the performance of approximately 93% of the guideline's recommended actions, when using the DSS mode. With respect to correctness, approximately 94.5% of the clinicians' decisions in the non-DSS mode were correct. However, these included 68% of the actions that were correct but redundant, given the patient's data (e.g., repeating tests that had been performed), and 27% of the actions, which were necessary in the context of the GL and of the given scenario. Only 5.5% of the decisions were definite errors. In the DSS mode, 94% of the clinicians' decisions were correct, which included 3% that were correct but redundant, and 91% of the actions that were correct and necessary in the context of the GL and of the given scenario. Only 6% of the DSS-mode decisions were erroneous. The DSS was assessed by the clinicians as potentially useful. Support from the GL-based DSS led to uniformity in the quality of the decisions, regardless of the particular clinician, any particular clinical scenario, any particular decision point, or any decision type within the scenarios. Using the DSS dramatically enhances completeness (i.e., performance of guideline-based recommendations) and seems to prevent the performance of most of the redundant actions, but does not seem to affect the rate of performance of incorrect actions. The redundancy rate is enhanced by similar recent findings in recent studies. Clinicians mostly find this support to be potentially useful for their daily practice. A continuous-care GL-based DSS, such as the Picard DSS engine, has the potential to prevent most errors of omission by ensuring uniformly high quality of clinical decision making (relative to a GL-based norm), due to the increased adherence (i.e., completeness) to the GL, and most of the errors of commission that increase therapy costs, by reducing the rate of redundant actions. However, to prevent clinical errors of commission, the DSS needs to be accompanied by additional modules, such as automated control of the quality of the physician's actual actions.


Why was PERV not transmitted during preclinical and clinical xenotransplantation trials and after inoculation of animals?
Porcine endogenous retroviruses (PERVs) are present in the genome of all pigs, they infect certain human cells and therefore pose a special risk for xenotransplantation using pig cells, tissues and organs. Xenotransplantation is being developed in order to alleviate the reduced availability of human organs. Despite the fact that PERVs are able to infect certain human cells and cells from other species, transmission of PERVs has not been observed when animals (including non-human primates) were inoculated with PERV preparations or during preclinical xenotransplantations. The data indicate that PERVs were not transmitted because they were not released from the transplant or were inhibited by intracellular restriction factors and innate immunity in the recipient. In a single study in guinea pigs, a transient PERV infection and anti-PERV antibodies were described, indicating that in this case at least, the immune system may also have been involved.


[Using biologic markers for evaluation of environmental pollution with metals].
Contemporary state of analytic basis enables to use human biologic materials adequately reflecting present ecologic situation for social hygienic monitoring of environmental pollution with heavy metals. To conduct biologic monitoring, regional databases on background metals content of human biologic materials are required. Regional data on elements contents of human body helps precise evaluation of health risk in exposure to environmental hazards.


Neighborhood socioeconomic status, race, and mortality in young adult dialysis patients.
Young blacks receiving dialysis have an increased risk of death compared with whites in the United States. Factors influencing this disparity among the young adult dialysis population have not been well explored. Our study examined the relation of neighborhood socioeconomic status (SES) and racial differences in mortality in United States young adults receiving dialysis. We merged US Renal Data System patient-level data from 11,027 black and white patients ages 18-30 years old initiating dialysis between 2006 and 2009 with US Census data to obtain neighborhood poverty information for each patient. We defined low SES neighborhoods as those neighborhoods in U.S. Census zip codes with ≥20% of residents living below the federal poverty level and quantified race differences in mortality risk by level of neighborhood SES. Among patients residing in low SES neighborhoods, blacks had greater mortality than whites after adjusting for baseline demographics, clinical characteristics, rurality, and access to care factors. This difference in mortality between blacks and whites was significantly attenuated in higher SES neighborhoods. In the United States, survival between young adult blacks and whites receiving dialysis differs by neighborhood SES. Additional studies are needed to identify modifiable factors contributing to the greater mortality among young adult black dialysis patients residing in low SES neighborhoods.


Rotavirus, adenovirus, and non-viral enteropathogens in diarrhoea.
The aetiology of rotavirus and adenovirus in acute gastroenteritis was studied in a prospective series that comprised 283 children admitted consecutively with diarrhoea during a 1-year period. Rotavirus was associated in 49% of the cases by solid-phase radioimmunoassay and electron microscopical examination of stool specimens, or by serology. Adenovirus was detected by radioimmunoassay in the stool specimens of 29 (11%) patients, including 8 cases of possible dual infection with rotavirus. Rotavirus infections showed a typical age distribution and seasonal clustering between January and June, whereas the adenovirus-associated cases did not form a distinctive subgroup. Enteropathogenic bacteria were found in 10% of cases, and were nearly as common in association with rotavirus infection as not. Respiratory symptoms accompanied diarrhoea in 34% of the patients with rotavirus and in 25% of those with neither rotavirus nor adenovirus. Therefore we could not confirm the existence of a 'rotavirus syndrome', nor could we confirm an association of respiratory symptoms with rotavirus infection. Use of antibiotics before the onset of diarrhoea was more common among those with non-viral diarrhoea (23%) than in the rotavirus group (13%). Rotavirus infections appeared to be common among cases of 'antibiotic-induced' diarrhoea.


Characterizing rheological properties and microstructure of thioester networks during degradation.
Covalent adaptable networks are designed for applications including cell and drug delivery and tissue regeneration. These applications require network degradation at physiological conditions and on a physiological timescale with microstructures that can: (1) support, protect and deliver encapsulated cells or molecules and (2) provide structure to surrounding tissue. Due to this, the evolving microstructure and rheological properties during scaffold degradation must be characterized. In this work, we characterize degradation of covalent adaptable poly(ethylene glycol) (PEG)-thioester networks with different amounts of excess thiol. Networks are formed between PEG-thiol and PEG-thioester norbornene using photopolymerization. These networks are adaptable because of a thioester exchange reaction that takes place in the presence of excess thiol. We measure degradation of PEG-thioester networks with L-cysteine using multiple particle tracking microrheology (MPT). MPT measures the Brownian motion of fluorescent probe particles embedded in a material and relates this motion to rheological properties. Using time-cure superposition (TCS), we characterize the microstructure of these networks at the gel-sol phase transition by calculating the critical relaxation exponent, n, for each network with different amounts of excess thiol. Based on the measured n values, networks formed with 0% and 50% excess thiol are tightly cross-linked and elastic in nature. While networks formed with 100% excess are similar to ideal, percolated networks, which have equal viscous and elastic components. MPT measurements during degradation of these networks also measure a non-monotonic increase in probe motility. We hypothesize that this is network rearrangement near the phase transition. We then measure macroscopic material properties including the equilibrium modulus and stress relaxation. We measure a trend in bulk network properties that agrees with the values of n. Elastic modulus and stress relaxation measurements show that networks with 50% excess thiol are more elastic compared to the other two networks. As the amount of excess thiol is increased from 0% to 50%, the networks become more elastic. Further increasing excess thiol to 100% reduces the elastically effective cross-links. We hypothesize that these properties are due to network non-idealities, resulting in networks with 50% excess thiol that are more elastic. This work characterizes dynamic rheological properties during degradation, which mimics processes that could occur during implantation. This work provides information that can be used in the future design of implantable materials enabling both the rheological properties and timescale of degradation to be specified.


Morphological and biochemical changes in the gills of Tilapia (Oreochromis mossambicus) to ambient cadmium exposure.
Tilapia, Oreochromis mossambicus, were reared in freshwater and exposed to 40, 80 and 160 ppb ambient cadmium for a period of 7 days to investigate the effects of short-term Cd exposure on the cellular function of gill chloride cells. Gills were sampled after 3 and 7 days exposure. The accumulated Cd concentration in gills was analyzed using a graphite furnace atomic absorption spectrophotometer while the morphological changes of pavement and chloride cells were examined using scanning and transmission electron microscopes. Gill Cd concentration was significantly increased in samples (10.03-44.36 ppb) at 3 and 7 days exposure. Accompanying this was an augmentation of microridge in pavement cells, and an increase in the density (1964-3603 /mm(2)) and apical membrane area (11.57-46.32 µm(2)) in chloride cells, indicating an adaptational modification of the cell morphology in assisting gaseous transfer and Ca(2+) uptake, respectively. However, biochemical analyses of the gill tissues enumerated a decrease in both the activities of alkaline and high-affinity Ca(2+)-ATPases. This indicated a reduction in the Ca(2+)-transport capacity per unit chloride cell, suggesting chloride cells being the primary target of Cd which subsequently lead to fish hypocalcemia.


Prognostic factors in patients with terminal stage lung cancer.
Lung cancer is the leading cause of cancer-related death.(1) Accurate prediction of survival in the terminal stage is important, because it may help patients make a rational decision. Although several prognostic scores have been described as effective indicators of outcome, these scores were intended for patients with other types of cancers. There is no prognostic score for patients with terminal-stage lung cancer. The aim of this study was to determine prognostic factors for patients with terminal-stage lung cancer. Patients in our palliative care unit (PCU) were selected retrospectively and divided into two independent groups, training and testing. Univariate and multivariate analyses were performed on data from the training group to detect independent prognostic factors, while data from patients in the testing group were analyzed to validate whether these prognostic factors predicted near-term death. Ninety-three patients (69 in the training group and 24 in the testing group) were included in the analyses. Multivariate analysis showed that fatigue, anorexia, desaturation, hyponatremia, and hypoalbuminemia were independent prognostic factors in the training group. Mean survival time in patients who had more than three of these five factors was 9.2±2.6 days (p=0.012). In the testing group, the presence of more than three of these five factors predicted death within two weeks, with a sensitivity of 100% and specificity of 75%. This study revealed that fatigue, anorexia, desaturation, hyponatremia, and hypoalbuminemia may be short-term prognostic factors in terminally ill lung cancer patients. In particular, the presence of more than three of these factors predicted death within two weeks.


Comparison of three restriction endonucleases in IS1245-based RFLP typing of Mycobacterium avium.
IS1245-based restriction fragment length polymorphism (RFLP) analysis has been proposed recently for molecular typing of Mycobacterium avium isolates. As there is no standardised method with respect to the optimal restriction enzyme, three restriction endonucleases were tested for analysis of 17 human isolates. The restriction endonucleases, selected on the basis of the physical maps of IS1245 and of the highly homologous IS1311, were BsaAI, that cleaves IS1245, PvuII, that cleaves IS1311, and NruI, that cleaves both IS1245 and IS1311. All the restriction endonucleases yielded polymorphic and complex RFLP patterns. However, BsaAI- and NruI-generated bands were more evenly distributed and easier to detect than PvuII-generated bands, most of which clustered in a narrow zone of the fingerprint. In some cases, DNA digestion with BsaAI or NruI yielded probe-specific restriction fragments of molecular size lower than expected. Moreover, digestion with NruI, which was expected to generate the highest numbers of bands in all the isolates, yielded fewer bands than were obtained with BsaAI or PvuII in 14 and 5 isolates, respectively. These findings might suggest the existence of unidentified IS1245-related insertion element(s) in M. avium isolates. Computer analysis of the IS1245-based RFLP patterns of M. avium isolates showed that the restriction endonucleases were capable, although with minor differences, of defining distinct banding patterns and clusters of identical or highly related isolates, thus confirming IS1245-based RFLP analysis as a useful technique for epidemiological studies.


Cprp-An Unusual, Repetitive Protein Which Impacts Pleuromutilin Biosynthesis in the Basidiomycete Clitopilus passeckerianus.
Interrogation of an EST database for Clitopilus passeckerianus identified a putative homolog to the unusual stress response gene from yeast; ddr48, as being upregulated under pleuromutilin production conditions. Silencing of this gene, named cprp, produced a population of transformants which demonstrated significantly reduced pleuromutilin production. Attempts to complement a Saccharomyces cerevisiae ddr48 mutant strain (strain Y16748) with cprp were hampered by the lack of a clearly identifiable mutant phenotype, but interestingly, overexpression of either ddr48 or cprp in S. cerevisiae Y16748 led to a conspicuous and comparable reduction in growth rate. This observation, combined with the known role of DDR48 proteins from a range of fungal species in nutrient starvation and stress responses, raises the possibility that this family of proteins plays a role in triggering oligotrophic growth. Localization studies via the production of a Cprp:GFP fusion protein in C. passeckerianus showed clear localization adjacent to the hyphal septa and, to a lesser extent, cell walls, which is consistent with the identification of DDR48 as a cell wall-associated protein in various yeast species. To our knowledge this is the first study demonstrating that a DDR48-like protein plays a role in the regulation of a secondary metabolite, and represents the first DDR48-like protein from a basidiomycete. Potential homologs can be identified across much of the Dikarya, suggesting that this unusual protein may play a central role in regulating both primary and secondary metabolism in fungi.


Interview: partnering with the industry: the Olympic laboratory model.
David A Cowan and Campbell P Barker speak to Ryan De Vooght-Johnson at Bioanalysis in May 2012 about the partnership between industry and academia for the setup and running of the doping-control laboratory for the London 2012 Olympic Games. David A Cowan is Director of the London 2012 anti-doping laboratory as well as Director of the King's College London Drug Control Centre, the UK's only WADA-accredited anti-doping laboratory, and Head of the Department of Forensic Science and Drug Monitoring. Cowan co-founded the Drug Control Centre in 1978 and became its Director in 1990. He has published extensively in the field of pharmaceutical analysis, especially as it relates to detecting drug administration in sport, and was awarded a personal chair in pharmaceutical toxicology in 1996. Cowan became Head of the Department of Forensic Science and Drug Monitoring at King's College London in 2002. He has served on a number of national and international committees, including the Council of Europe Working Party Investigating Drug Abuse in Sport that led to the first World Anti-Doping Convention, the Laboratory Representative on the International Olympic Committee's Medical Commission, and WADA's Laboratory Accreditation Subcommittee. He is a member of the Crippen Club for Distinguished Toxicologists. In 1998 he was awarded the IOC Trophy for Sport Ethics by the BOA. He was a founding member of the World Association of Anti-Doping Scientists and became its first President serving on its Executive Board between 2001 and 2004. He was a Visiting Laboratory Director at the Salt Lake City Winter Olympic Games 2002, where the first novel erythropoiesis-stimulating protein (NESP) positive was discovered. He was also a senior advisory scientist at both the Turin Winter Olympic Games in 2006 and the Beijing Olympic Games in 2008. He was also a member of the IOC Medical Commission for the Sydney Olympic Games in 2000 and the Vancouver Winter Olympic Games in 2010. The Drug Control Centre undertook the sample analysis during the 2002 Manchester Commonwealth Games and Cowan was Co-Director of the laboratory for the Commonwealth Games in Kuala Lumpur in 1998. Cowan, who has directed the laboratory at King's College London for many years, was a member of the bid team making the presentation to the International Olympic Committee in February 2005. Campbell P Barker has been leading GlaxoSmithKline's delivery of the London 2012 laboratory services since September 2009. Prior to that time he was Director of Global Strategic Projects in GlaxoSmithKline's Consumer Healthcare business from 2006, and from 1994 to 2006 he worked in R&D for Procter & Gamble. Barker holds a PhD in chemistry from the University of Durham.


Interventions to reduce the bacterial load in recycled broiler litter.
Two experiments were undertaken to evaluate the bacterial load in recycled litter between broiler flocks following addition of quicklime (T1), windrowing (T2), shallow fermentation (T3), and control (no intervention, T4). The first experiment was developed in field conditions in which the broiler houses were accompanied by 6 consecutive flocks and the effect of the treatments was assessed on enterobacteria and aerobic mesophiles. The second experiment was conducted in an experimental broiler house with recycled litter for assessment of Salmonella Enteritidis phage type 4 (SE PT4). In the field study, T3 presented the best results in reducing enterobacteria in broiler litter in relation to the other treatments, with the highest reduction occurring in the first 3 flocks, tending to stabilization from the fourth flock onward for all the treatments assessed. From the third to sixth flocks, enterobacteria level at the end of the treatments (d 12) was lower than the average in the fresh litter, except in T4. All treatments reduced aerobic mesophiles throughout the flocks, where T2 showed the highest reduction. The percentage of dry matter in the broiler litter diminished in T4 and increased in T3 over the course of the flocks. In the second experiment, the drop in the SE PT4 level in the broiler litter first occurred in T2 and T3. However, all the treatments except for T4 eliminated SE PT4 within 12 d. The temperature of the broiler litter in T2 was higher in relation to the other treatments. The results show that litter treatment prior to reutilization by the successive broiler flock is required to reduce the level of residual bacteria. The fermentative treatments (T2 and T3) were found to be superior to the others in terms of reducing the bacterial load, with shallow fermentation standing out with the highest reduction of enterobacteria and equivalent SE PT4 elimination when compared to windrowing.


Defects in lamin B1 expression or processing affect interphase chromosome position and gene expression.
Radial organization of nuclei with peripheral gene-poor chromosomes and central gene-rich chromosomes is common and could depend on the nuclear boundary as a scaffold or position marker. To test this, we studied the role of the ubiquitous nuclear envelope (NE) component lamin B1 in NE stability, chromosome territory position, and gene expression. The stability of the lamin B1 lamina is dependent on lamin endoproteolysis (by Rce1) but not carboxymethylation (by Icmt), whereas lamin C lamina stability is not affected by the loss of full-length lamin B1 or its processing. Comparison of wild-type murine fibroblasts with fibroblasts lacking full-length lamin B1, or defective in CAAX processing, identified genes that depend on a stable processed lamin B1 lamina for normal expression. We also demonstrate that the position of mouse chromosome 18 but not 19 is dependent on such a stable nuclear lamina. The results implicate processed lamin B1 in the control of gene expression as well as chromosome position.


[Application of catalytic model in sero-epidemiology of schistosomiasis].
The seroepidemiological data from two schistosomiasis japonica areas with different transmission levels was analysed using a Catalytic Model. The age-specific distribution of IgG antibody in the population of the two investigated settings was analysed with simple catalytic model, the curve equations are Y(A) = 0.876 (1--e-0.048t) and Y(B) = 5.71 (1--e-0.003t) respectively in the heavy- and mild-endemic areas. The age-specific distribution of IgM antibody was analysed with two-stage catalytic model, using a curve equations Y(A) = 2.13 (e-0.032t--e-0.047t) and Y(B) = 0.19 (e-0.012t--e-0.075t) respectively in both heavy-and mild-endemic areas. Based on the result of stool examination concurrently carried out with sera detection and the endemic situation in recent years, the results mentioned above were eligible for the analysis of seroepidemiological data on schistosomiasis to reveal the endemic situation, tendency and the characteristic of age-specific distribution, and in particular using a two-stage catalytic model to analyse IgM antibody. It could also be used for studies on endemic factors, evaluation on the efficiency of control programmes and for surveillance of endemicy.


[Candidemia and invasive candidiasis approach in critically ill patients: role of the echinocandins].
Invasive infections caused by Candida spp. in critically ill patients may significantly worsen their prognosis, so it is of great importance to establish an early detection and a suitable therapeutic strategy. The objective of this study was to define the differential role of echinocandins in treating certain critical patient profiles. A scientific committee of 9 experts in infectious diseases, critical care, microbiology, and hospital pharmacy reviewed the existing evidence on the treatment of candidemia and invasive candidiasis in critically ill patients. After that, a questionnaire with 35 items was elaborated to be agreed by 26 specialists in the aforementioned disciplines using a modified Delphi method. After two rounds of evaluation, a consensus was reached in terms of agreement in 66% of the items. Some of the consensuses achieved included: it is not necessary to adjust the dose of echinocandins during renal replacement therapy; the echinocandins are the empirical and/or directed treatment of choice for candidemia and invasive candidiasis associated with biofilms; these drugs may be used in the antifungal prophylaxis of high-risk liver transplantation. In the absence of additional clinical data, it should be noted that micafungin is the echinocandin with the most available scientific evidence. The experts consulted showed a high degree of agreement on some of the most controversial aspects regarding the management of candidemia and invasive candidiasis in critical patients, which could inform of practical recommendations for their treatment.


Radiotherapy, Bisphosphonates and Surgical Stabilization of Complete or Impending Pathologic Fractures in Patients with Metastatic Bone Disease.
To report the treatment outcomes of patients with metastatic bone disease with complete or impending pathologic fractures, who were treated with postoperative radiotherapy (RT), bisphosphonates or both after orthopedic stabilization. We retrospectively evaluated the results of RT, bisphosphonates or both after orthopedic stabilization for complete or impending pathologic fractures in 72 patients with skeletal metastases. After surgery, 32 patients (44%) were treated with RT alone (group 1), 31 patients (43%) were treated with RT and bisphosphonates (group 2) and 9 (13%) patients were treated with bisphosphonates (group 3), respectively. Patients were treated with a median dose of 30Gy (30-40 Gy/2-3Gy per fraction).The local tumor progression, pain progression and need for re-operation or re-radiotherapy were assessed from patients' medical records. Median follow-up time was 9 months. Median overall survival time was 14 months (95% CI: 12-17). Secondary surgical intervention at the same location was necessary in 1 patient of group 1 (2%), 2 patients of group 2(5%) and 2 patients of group 3 (15%), respectively (p=0.097). Local tumor progress was observed in 3 patients of group 1 (9%), 2 patients of group 2 (7%) and 4 patients in group 3 (44%), respectively (p=0.021). Local pain progress was observed in 19%, 16% and 67% of the same groups (p=0.011). Our data confirm the efficacy and necessity of postoperative RT after orthopedic stabilization for metastatic bone disease to control the local disease. Bisphosphonates do not obviate the need for RT in the management of bone metastases after surgical stabilization. The combined treatment might lead to a better local tumor and pain control.


Increasing Mass Transfer Resistance of MOFs as a Reverse Tuning Strategy to Achieve High-Resolution Gas Chromatographic Separation.
In separation science, precise control and regulation of the MOF stationary phase are crucial for achieving a high separation performance. We supposed that increasing the mass transfer resistance of MOFs with excessive porosity to achieve a moderate mass transfer resistance of the analytes is the key to conducting the MOF stationary phase with a high resolution. Three-dimensional UiO-67 (UiO-67-3D) and two-dimensional UiO-67 (UiO-67-2D) were chosen to validate this strategy. Compared with UiO-67-3D with overfast mass transfer and low retention, the reduced porosity of UiO-67-2D increased the mass transfer resistance of analytes in reverse, resulting in improved separation performance. Kinetic diffusion experiments were conducted to verify the difference in mass transfer resistance of the analytes between UiO-67-3D and UiO-67-2D. In addition, the optimization of the UiO-67-2D thickness for separation revealed that a moderate diffusion length of the analytes is more advantageous in achieving the equilibrium of absorption and desorption.


Fibrinogen surface distribution correlates to platelet adhesion pattern on fluorinated surface-modified polyetherurethane.
In previous work, it had been shown that platelet adhesion could be reduced by fluorinating surfaces with oligomeric fluoropolymers, referred to as surface-modifying macromolecules (SMMs). In the current study, two in vitro blood-contacting experiments were carried out on a polyetherurethane modified with three different SMMs in order to determine if altered platelet adhesion levels could be related to the pattern of adsorbed protein and more specifically to the manner in which fibrinogen (Fg) distribution occurs at the surface. In the first experiment, the materials were placed in whole human blood and the adherent platelets were viewed with high-resolution scanning electron microscopy (SEM). In a second experiment, the materials were incubated with human plasma with the absence of platelets. The plasma contained 5% fluorescent-Fg. The materials were then viewed with a fluorescence microscope and images were collected to define the distribution of high-density fluorescent-Fg areas. The SEM and fluorescent-Fg images were imported to Image Pro Plus imaging software to measure the area, length and circularity and a bivariate correlation test was conducted between the two sets of data. For area and length morphology parameters, there were high and significant correlations (r > 0.9, p < 0.05) between the platelets and Fg aggregates. The data suggest that the Fg distribution may serve as a predictor of platelet morphology/activation and provides insight into the non-thrombogenic character of biomaterials containing the fluorinated SMMs.


Examination of disease severity in systemic vasculitis from the novel perspective of damage using the vasculitis damage index (VDI).
Assessment of disease severity in systemic vasculitis encompasses mortality, which is now uncommon, and morbidity, which is increasing in significance. Morbidity includes permanent scars or damage, an evolving concept offering a novel perspective which may be particularly valuable in chronic disease. We have developed a method for assessing damage in systemic vasculitis, but the relationship between damage and disease severity was unknown. Therefore, we examined whether the number of items of damage or the pattern of damage varied with the severity of systemic vasculitis. We established the characteristics of severe disease by examining fatal vasculitis as an example of the most severe disease possible. We then showed that more damage occurred in fatal vasculitis, more systems were damaged, and critical damage akin to organ failure was more common in fatal than non-fatal vasculitis. These observations were reproduced in specific diagnostic groups, namely classical Wegener's granulomatosis and systemic rheumatoid vasculitis. Thus, severe disease was characterized by many items of damage, multisystem damage and critical damage. This pattern of damage was also seen in a subgroup of patients with non-fatal vasculitis, who also have severe disease.


Auditory cortical responses in patients with cochlear implants.
Currently, the most commonly used electrophysiological tests for cochlear implant evaluation are Averaged Electrical Voltages (AEV), Electrical Advisory Brainstem Responses (EABR) and Neural Response Telemetry (NRT). The present paper focuses on the study of acoustic auditory cortical responses, or slow vertex responses, which are not widely used due to the difficulty in recording, especially in young children. Aims of this study were validation of slow vertex responses and their possible applications in monitoring postimplant results, particularly restoration of hearing and auditory maturation. In practice, the use of tone-bursts, also through hearing aids or cochlear implants, as in slow vertex responses, allows many more frequencies to be investigated and louder intensities to be reached than with other tests based on a click as stimulus. Study design focused on latencies of N1 and P2 slow vertex response peaks in cochlear implants. The study population comprised 45 implant recipients (aged 2 to 70 years), divided into 5 different homogeneous groups according to chronological age, age at onset of deafness, and age at implantation. For each subject, slow vertex responses and free-field auditory responses (PTAS) were recorded for tone-bursts at 500 and 2000 Hz before cochlear implant surgery (using hearing aid amplification) and during scheduled sessions at 3rd and 12th month after implant activation. Results showed that N1 and P2 latencies decreased in all groups starting from 3rd through 12th month after activation. Subjects implanted before school age or at least before age 8 yrs showed the widest latency changes. All subjects showed a reduction in the gap between subjective thresholds (obtained with free field auditory responses) and objective thresholds (obtained with slow vertex responses), obtained in presurgery stage and after cochlear implant. In conclusion, a natural evolution of neurophysiological cortical activities of the auditory pathway, over time, was found especially in young children with prelingual deafness and implanted in preschool age. Cochlear implantation appears to provide hearing restoration, demonstrated by the sharp reduction of the gap between subjective free field auditory responses and slow vertex responses threshold obtained with hearing aids vs. cochlear implant.


Dietary Supplement Use during Preconception: The Australian Longitudinal Study on Women's Health.
Worldwide, dietary supplement use among reproductive aged women is becoming increasingly common. The aim of this study was to investigate dietary supplement use among Australian women during preconception. Self-reported data were collected prospectively for the Australian Longitudinal Study on Women's Health (ALSWH). The sample included 485 women aged 31-36 years, with supplement data, classified as preconception when completing Survey 5 of the ALSWH in 2009. Frequency and contingency tables were calculated and Pearson's chi-square test for associations between demographic variables and supplementation status was performed. Sixty-three per cent of women were taking at least one dietary supplement during preconception. Multiple-micronutrient supplements were the most commonly reported supplement (44%). Supplements containing folic acid and iodine were reported by 51% and 37% of preconception women, respectively. Folic acid (13%), omega-3 fatty acids (11%), vitamin C (7%), B vitamins (4%), iron (3%), and calcium (3%) were the most common single nutrients supplemented during preconception. Women trying to conceive, with no previous children, and born outside Australia were more likely to take dietary supplements. In Australia, dietary supplement use during preconception is relatively high. However, supplementation of recommended nutrients, including folic acid and iodine, could be improved.


Synthetic prions with novel strain-specified properties.
Prions are infectious proteins that possess multiple self-propagating structures. The information for strains and structural specific barriers appears to be contained exclusively in the folding of the pathological isoform, PrP(Sc). Many recent studies determined that de novo prion strains could be generated in vitro from the structural conversion of recombinant (rec) prion protein (PrP) into amyloidal structures. Our aim was to elucidate the conformational diversity of pathological recPrP amyloids and their biological activities, as well as to gain novel insights in characterizing molecular events involved in mammalian prion conversion and propagation. To this end we generated infectious materials that possess different conformational structures. Our methodology for the prion conversion of recPrP required only purified rec full-length mouse (Mo) PrP and common chemicals. Neither infected brain extracts nor amplified PrP(Sc) were used. Following two different in vitro protocols recMoPrP converted to amyloid fibrils without any seeding factor. Mouse hypothalamic GT1 and neuroblastoma N2a cell lines were infected with these amyloid preparations as fast screening methodology to characterize the infectious materials. Remarkably, a large number of amyloid preparations were able to induce the conformational change of endogenous PrPC to harbor several distinctive proteinase-resistant PrP forms. One such preparation was characterized in vivo habouring a synthetic prion with novel strain specified neuropathological and biochemical properties.


Vascular endothelial growth factor and soft tissue sarcomas: tumor expression correlates with grade.
Vascular endothelial growth factor (VEGF), an endothelial-specific mitogen overexpressed in various epithelial malignancies is thought to be a potent regulator of angiogenesis. We hypothesized that some soft tissue sarcomas, due to their high propensity for hematogenous metastases (1) would overexpress VEGF, (2) that the degree of expression may represent a significant biologic predictor for disease-specific survival, and (3) that recurrent tumor would express as high or higher VEGF compared with the primary tumor. Selected paraffin-embedded tissue of surgical specimens from 79 patients with soft tissue sarcomas, treated between 1989 and 1995 were stained with a rabbit polyclonal anti-VEGF antibody at a concentration of 2 microg/ml. Slides were assessed for VEGF expression as high or low by two investigators blinded to the clinicopathologic data. Twelve patients had VEGF expression of their primary tumors, and their recurrent tumors were compared. The Fishers' exact test assessed for differences in VEGF expression; survival analyses were performed according to the methods of Kaplan and Meier. Seventy-eight percent (29 of 37) of patients who died of disease had high VEGF expression. However, VEGF expression was not an independent predictor of either overall or disease-free survival. Tumor grade correlated with VEGF expression significantly. For the low-grade tumors, 7 of 13 expressed low VEGF, whereas for high-grade tumors, 53 of 66 expressed high VEGF (P = .016). Seven of the 12 paired tumor samples expressed identical VEGF immunostaining. The majority of high-grade soft tissue sarcomas in this study have high intensity VEGF expression. This finding may provide useful information on individual soft tissue sarcomas and offer the basis for therapeutic and biologic targeting in high-risk patients using anti-angiogenesis strategies. However, in our analysis, after accounting for tumor grade, VEGF does not seem to be an independent predictor of clinical outcome.


NICTABA and UDA, two GlcNAc-binding lectins with unique antiviral activity profiles.
This study aimed to assess the antiviral properties of a unique lectin (NICTABA) produced by the tobacco plant, Nicotiana tabacum. Cellular assays were used to investigate the antiviral activity of NICTABA and Urtica dioica agglutinin (UDA). Surface plasmon resonance (SPR) studies were performed to study the sugar specificity and the interactions of both lectins with the envelope glycoproteins of HIV-1. The N-acetyl-d-glucosamine (GlcNAc)-binding lectins exhibited broad-spectrum activity against several families of enveloped viruses including influenza A/B, Dengue virus type 2, herpes simplex virus types 1 and 2 and HIV-1/2. The IC50 of NICTABA for various HIV-1 strains, clinical isolates and HIV-2 assessed in PBMCs ranged from 5 to 30 nM. Furthermore, NICTABA inhibited syncytium formation between persistently HIV-1-infected T cells and uninfected CD4+ T lymphocytes and prevented DC-SIGN-mediated HIV-1 transmission to CD4+ target T lymphocytes. However, unlike many other antiviral carbohydrate-binding agents (CBAs) described so far, NICTABA did not block HIV-1 capture to DC-SIGN+ cells and it did not interfere with the binding of the human monoclonal antibody 2G12 to gp120. SPR studies with HIV-1 envelope glycoproteins showed that the affinity of NICTABA for gp120 and gp41 was in the low nanomolar range. The specific binding of NICTABA to gp120 could be prevented in the presence of a GlcNAc trimer, but not in the presence of mannose trimers. NICTABA displayed no antiviral activity against non-enveloped viruses. Since CBAs possess a high genetic barrier for the development of viral resistance and NICTABA shows a broad antiviral activity profile, this CBA may qualify as a potential antiviral candidate with a pleiotropic mode of action aimed at targeting the entry of enveloped viruses.


[The formation of motor units in vertebrates (a histophysiological analysis)].
In motor unit anlage successive formation of morphophysiological and biochemical components of system-forming units (histophysiological compartments) occurs. Simultaneously functional differentiation of motor units takes place, based on morphological, biochemical and functional heterogeneity of nervous, muscular and synaptic motor unit components. Intrinsic genetic programmes of main nervous and skeletal muscular tissue differons are the leading ones in early stages of embryogenesis. Morphofunctional base for realization of neurogenic control of skeletal musculature fibres histochemical differentiation forms in muscle anlage. Genetic programmes of two tissues development come to direct (contact) interaction through outer regions of cell membranes: motor neurons participation in skeletal muscular tissue histogenesis, especially on the stage of muscular tubules differentiation into muscular fibres of certain histophysiological type is essential. Most complicated changes are observed in postsynaptic membrane area. This allows to emphasize the development of the symphast nucleo-sarcoplasmic compartment. In further, adaptive changes and morphological manifestations of skeletal muscular tissue genetic programme realization arise in myogenesis with motor neurons role progressively increasing and becoming the predominant one.


Viscoelastic Properties Measurement of Human Lymphocytes by Atomic Force Microscopy Based on Magnetic Beads Cell Isolation.
Cell mechanics has been proved to be an effective biomarker for indicating cellular states. The advent of atomic force microscopy (AFM) provides an exciting instrument for measuring the mechanical properties of single cells. However, current AFM single-cell mechanical measurements are commonly performed on cell lines cultured in vitro which are quite different from the primary cells in the human body. Investigating the mechanical properties of primary cells from clinical environments can help us to better understand cell behaviors. Here, by combining AFM with magnetic beads cell isolation, the viscoelastic properties of human primary B lymphocytes were quantitatively measured. B lymphocytes were isolated from the peripheral blood of healthy volunteers by density gradient centrifugation and CD19 magnetic beads cell isolation. The activity and specificity of the isolated cells were confirmed by fluorescence microscopy. AFM imaging revealed the surface topography and geometric parameters of B lymphocytes. The instantaneous modulus and relaxation time of living B lymphocytes were measured by AFM indenting technique, showing that the instantaneous modulus of human normal B lymphocytes was 2-3 kPa and the relaxation times were 0.03-0.06 s and 0.35-0.55 s. The differences in cellular visocoelastic properties between primary B lymphocytes and cell lines cultured in vitro were analyzed. The study proves the capability of AFM in quantifying the viscoelastic properties of individual specific primary cells from the blood sample of clinical patients, which will improve our understanding of the behaviors of cells in the human body.


Fibroblast growth factor 5 protects against spinal cord injury through activating AMPK pathway.
Excessive productions of inflammatory cytokines and free radicals are involved in spinal cord injury (SCI). Fibroblast growth factor 5 (FGF5) is associated with inflammatory response and oxidative damage, and we herein intend to determine its function in SCI. Lentivirus was instilled to overexpress or knockdown FGF5 expression in mice. Compound C or H89 2HCl were used to suppress AMP-activated protein kinase (AMPK) or protein kinase A (PKA), respectively. FGF5 level was significantly decreased during SCI. FGF5 overexpression mitigated, while FGF5 silence further facilitated inflammatory response, oxidative damage and SCI. Mechanically, FGF5 activated AMPK to attenuate SCI in a cAMP/PKA-dependent manner, while inhibiting AMPK or PKA with pharmacological methods significantly abolished the neuroprotective effects of FGF5 against SCI. More importantly, serum FGF5 level was decreased in SCI patients, and elevated serum FGF5 level often indicate better prognosis. Our study identifies FGF5 as an effective therapeutic and prognostic target for SCI.


Selected parameters of the corneal deformation in the Corvis tonometer.
Contemporary ophthalmology knows many methods of measuring intraocular pressure, namely the methods of non-contact and impression applanation tonometry. In non-contact applanation tonometers, e.g. the Corvis, the corneal flattening is caused by an air puff. Image registration of the corneal deflection performed by a tonometer enables to determine other interesting biomechanical parameters of the eye, which are not available in the tonometer. The measurement of new selected parameters is presented in this paper. Images with an M × N × I resolution of 200 × 576 × 140 pixels were acquired from the Corvis device in the source recording format *.cst. A total of 13'400 2D images of patients examined routinely in the Clinical Department of Ophthalmology, in District Railway Hospital in Katowice, Poland, were analysed in accordance with the Declaration of Helsinki. A new method has been proposed for the analysis of corneal deflection images in the Corvis tonometer with the use of the Canny edge detection method, mathematical morphology methods and context-free operations. The resulting image analysis tool allows determination of the response of the cornea and the entire eyeball to an air puff. The paper presents the method that enables the measurement of the amplitude of curvature changes in the frequency range from 150 to 500 Hz and automatic designation of the eyeball movement direction. The analysis of these data resulted in 3 new features of dynamics of the eye reaction to an air puff. Classification of these features enabled to propose 4 classes of deformation. The proposed algorithm allows to obtain reproducible results fully automatically at a time of 5 s per patient using the Core i5 CPU M460 @ 2.5GHz 4GB of RAM. The paper presents the possibility of using a profiled algorithm of image analysis, proposed by the authors, to measure additional cornea deformation parameters. The new tool enables automatic measurement of the additional new parameters when using the Corvis tonometer. A detailed clinical examination based on this method will be presented in subsequent papers.


Prognostic Indicators of Non-Transection Nerve Injury and Vocal Fold Motion Impairment After Thyroid Surgery - Correlation Between Intraoperative Neuromonitoring Findings and Perioperative Voice Parameters.
In patients with recurrent laryngeal nerve (RLN) injury after thyroid surgery, unrecovered vocal fold motion (VFM) and subjective voice impairment cause extreme distress. For surgeons, treating these poor outcomes is extremely challenging. To enable early treatment of VFM impairment, this study evaluated prognostic indicators of non-transection RLN injury and VFM impairment after thyroid surgery and evaluated correlations between intraoperative neuromonitoring (IONM) findings and perioperative voice parameters. 82 adult patients had postoperative VFM impairment after thyroidectomy were enrolled. Demographic characteristics, RLN electromyography (EMG), and RLN injury mechanism were compared. Multi-dimensional voice program, voice range profile and Index of voice and swallowing handicap of thyroidectomy (IVST) were administered during I-preoperative; II-immediate, III-short-term and IV-long-term postoperative periods. The patients were divided into R/U Group according to the VFM was recovered/unrecovered 3 months after surgery. The patients in U Group were divided into U1/U2 Group according to total IVST score change was <4 and ≥4 during period-IV. Compared to R Group (42 patients), U Group (38 patients) had significantly more patients with EMG >90% decrease in the injured RLN (p<0.001) and thermal injury as the RLN injury mechanism (p=0.002). Voice parameter impairments were more severe in U Group compared to R Group. Compared to U1 group (19 patients), U2 Group (19 patients) had a significantly larger proportion of patients with EMG decrease >90% in the injured RLN (p=0.022) and thermal injury as the RLN injury mechanism (p=0.017). A large pitch range decrease in period-II was a prognostic indicator of a moderate/severe long-term postoperative subjective voice impairment. This study is the first to evaluate correlations between IONM findings and voice outcomes in patients with VFM impairment after thyroid surgery. Thyroid surgeons should make every effort to avoid severe type RLN injury (e.g., thermal injury or injury causing EMG decrease >90%), which raises the risk of unrecovered VFM and moderate/severe long-term postoperative subjective voice impairment. Using objective voice parameters (e.g., pitch range) as prognostic indicators not only enables surgeons to earlier identify patients with low voice satisfaction after surgery, and also enable implementation of interventions sufficiently early to maintain quality of life.


Protection of p-Coumaric acid against chronic stress-induced neurobehavioral deficits in mice via activating the PKA-CREB-BDNF pathway.
There is a body of evidence to suggest that chronic stress modulates neurochemical homeostasis, alters neuronal structure, inhibits neurogenesis and contributes to development of mental disorders. Chronic stress-associated mental disorders present common symptoms of cognitive impairment and depression with complex disease mechanisms. P-coumaric acid (p-CA), a natural phenolic compound, is widely distributed in vegetables, cereals and fruits. p-CA exhibits a wide range of health-related effects, including anti-oxidative-stress, anti-mutagenesis, anti-inflammation and anti-cancer activities. The current study aims to evaluate the therapeutic potential of p-CA against stress-associated mental disorders. We assessed the effect of p-CA on cognitive deficits and depression-like behavior in mice exposed to chronic restraint stress (CRS); we used network pharmacology, biochemical and molecular biological approaches to elucidate the underlying molecular mechanisms. CRS exposure caused memory impairments and depression-like behavior in mice; p-CA administration attenuated these CRS-induced memory deficits and depression-like behavior. Network pharmacology analysis demonstrated that p-CA was possibly involved in multiple targets and a variety of signaling pathways. Among them, the protein kinase A (PKA) - cAMP-response element binding protein (CREB) - brain derived neurotrophic factor (BDNF) signaling pathway was predominant and further characterized. The levels of PKA, phosphorylated CREB (pCREB) and BDNF were significantly lowered in the hippocampus of CRS mice, suggesting disruption of the PKA-CREB-BDNF signaling pathway; p-CA treatment restored the signaling pathway. Furthermore, CRS upregulated expression of proinflammatory cytokines in hippocampus, while p-CA reversed the CRS-induced effects. Our findings suggest that p-CA will offer therapeutic benefit to patients with stress-associated mental disorders.


Design optimization of a total knee replacement for improved constraint and flexion kinematics.
Total knee replacement (TKR) constraint and flexion range of motion can be limiting factors in terms of kinematics performance and cause for revision. These characteristics are closely related to the shape of the implant components. No previous studies have used a rigorous and systematic design optimization method to determine the optimal shape of TKR components. Previous studies have failed to define a quantifiable objective function for optimization, have not used any optimization algorithms, and have only considered a limited design space (4 or less design variables). This study addresses these limitations and determines the optimum shape of the femoral component and ultra high molecular weight polyethylene (UHMWPE) insert in terms of kinematics. The constraint characteristics with respect to those of the natural knee, the importance of the posterior cruciate ligament, and the flexion range of motion were all considered. The kinematics optimized design featured small femoral radii of curvature in the frontal and sagittal planes, but asymmetric with slightly larger radii of curvature for the lateral condyle. This condyle was also less conforming than the medial side. Compared to a commercially available TKR design, the kinematics performance (based on constraint and flexion range of motion) was improved by 81%, with constraint characteristics generally closer to those of the natural knee and a 12.6% increase in the flexion range of motion (up to 143°). The results yielded a new TKR design while demonstrating the feasibility of design optimization in TKR design.


Case Report: PsAPSASH syndrome: an alternative phenotype of syndromic hidradenitis suppurativa treated with the IL-17A inhibitor secukinumab.
Syndromic hidradenitis suppurativa (HS) is a form of symptom constellations, which differs from the familial and genetic form and comprises predominantly osteoarticular manifestations. Many forms include pyoderma gangrenosum and acne (PASH), pyogenic arthritis (PAPASH), spondyloarthritis (PASS) and psoriatic arthritis (PsaPASH) and are categorized in the autoinflammatory syndromes. anti-TNF-α and anti-IL-1a blockade are between the therapeutic approaches that improve skin symptoms and prevent permanent osteoarticular damage. This case report refers to the successful treatment of a mixed phenotype of the aforementioned symptoms using the IL-17A inhibitor secukinumab after initial treatment with adalimumab. The therapy improved both cutaneous and reported osteoarticular symptoms. Different approaches for these recalcitrant HS syndromes are essential in order to achieve long-term remission for those patients.


Studies on the cultivation of influenza virus in vitro.
Studies have been carried out since 1959 at the Coonoor Influenza Centre to devise a method of cultivating influenza virus in vitro which would be suitable for large-scale virus production. The authors report the successful cultivation of the virus in tissue cultures of chorioallantoic membrane on glass wool. The method described may be used equally satisfactorily for culture in volumes ranging from 1.0 ml to 350 ml, and is as sensitive as cultivation in eggs for the titration of different strains of influenza virus and their neutralizing antibodies. Relatively pure virus for vaccine production and complement-fixing antigen for diagnostic purposes can be produced in large volumes with ease and economy.


Exposure to e-cigarette TV advertisements among U.S. youth and adults, 2013-2019.
E-cigarette advertising has been shown to increase e-cigarette awareness and use. Although e-cigarette marketing in the early 2010s has been well-documented, little is known about how it has changed in recent years in response to the regulatory scrutiny from the FDA and the Congress to combat youth vaping epidemic. This study aims to examine the exposure to e-cigarette TV advertising among youth and adults in the U.S. from 2013 to 2019, overall and by media market and brand. Quarterly data on e-cigarette TV advertising exposure, measured by target rating points (TRPs), and expenditures from 2013 to 2019 were compiled from the StradegyTM of Kantar Media. Trends of quarterly e-cigarette advertising TRPs were reported by age group, market, and brand. Over the study period, overall exposure to e-cigarette TV advertising was higher among adults than among youth. E-cigarette advertising TRPs and expenditures were relatively stable, despite intermittent fluctuations, between 2013 Q1 and 2017 Q1 except for a one-time dip in 2015 Q3, followed by a sharp decline in 2017 Q2 and stayed low till the end of 2018. A resurgence of e-cigarette advertising TRPs occurred in 2019 Q1, led by the advertising from JUUL, Blu, and Vuse, which peaked in the third quarter of 2019, with quarterly TRPs reaching 316.8 for youth and 1,701.9 for adults, and quarterly advertising expenditure totaling $31 million. Significant variations, both over time and across media markets and brands, were observed for e-cigarette televised advertising between 2013 and 2019. Following a lull in TV advertising in 2017/18, major e-cigarette companies have substantially increased advertising of their products on American television since early 2019, resulting in a surge in exposure to e-cigarette advertising among both youth and adults. Our findings highlighted the importance of continued monitoring of e-cigarette advertising in the U.S.


Lack of radiation protection for endoscopists performing endoscopic retrograde cholangiopancreatography.
ERCP using fluoroscopy should be practiced with an adequate radiation protection. However, the awareness of gastrointestinal endoscopists to radiation protection was considered insufficient. In Korea, a country with a rapid increase the number of ERCP procedures, there is no data about radiation protection practices for gastrointestinal endoscopists. The purpose of this study was to investigate current clinical practices and the awareness on radiation protection in ERCP performing physicians in Korea. An anonymous questionnaire regarding radiation protection practices was mailed to 100 members of Korean Pancreatobiliary Association who was porforming ERCP. The questionnaire included ERCP volume of each endoscopist, use of protection devices such as apron, thyroid shield, lead glasses and any mobile shield for scattered radiation, and whether they monitored their own radiation exposure dosage. All respondents wore lead aprons during ERCP. While 52.5% of endoscopists answered that they always wear thyroid guards, 26.9% rarely or never wore it. Only 14% wore lead glasses during the procedure and 69% never wore it. The preparation rates of mobile shields or lead curtains were only 14% and 24%, respectively. Only 10% of endoscopists attached an X-ray badge and 66.7% never used it. Moreover, 75% of endoscopists responded that they did not monitor their own exposure dose to radiation during ERCP. The lack of radiation protection of ERCP endoscopists in Korea was seemed serious. Awareness of radiation hazard should be more concerned and educated in parallel with the preparation of radiation protection equipments.


Safety and feasibility of targeted agent combinations in solid tumours.
The plethora of novel molecular-targeted agents (MTAs) has provided an opportunity to selectively target pathways involved in carcinogenesis and tumour progression. Combination strategies of MTAs are being used to inhibit multiple aberrant pathways in the hope of optimizing antitumour efficacy and to prevent development of resistance. While the selection of specific agents in a given combination has been based on biological considerations (including the role of the putative targets in cancer) and the interactions of the agents used in combination, there has been little exploration of the possible enhanced toxicity of combinations resulting from alterations in multiple signalling pathways in normal cell biology. Owing to the complex networks and crosstalk that govern normal and tumour cell proliferation, inhibiting multiple pathways with MTA combinations can result in unpredictable disturbances in normal physiology. This Review focuses on the main toxicities and the lack of tolerability of some common MTA combinations, particularly where evidence of enhanced toxicity compared to either agent alone is documented or there is development of unexpected toxicity. Toxicities caused by MTA combinations highlight the need to introduce new preclinical testing paradigms early in the drug development process for the assessment of chronic toxicities resulting from such combinations.


Coexistence of sickle cell disease and systemic lupus erythematosus is associated with quantitative and qualitative impairments in circulating regulatory B cells.
The incidence of connective tissue diseases such as systemic lupus erythematous (SLE), in adult patients with sickle cell disease (SCD), appears to be increasing. The exact causes underlying this increased risk are still unknown, but a link with B regulatory (Breg) cells is possible as these cells suppress inflammatory responses, and maintain tolerance. Quantitative and qualitative analyses of circulating Breg cells were performed in a cohort of SCD patients with SLE, and their levels were correlated with key soluble mediators promoting autoreactive B cells. We demonstrated that levels of Breg cells were significantly decreased in SCD patients with SLE compared to patients with SCD only or healthy controls. Functional analysis of Breg cells from SCD patients with SLE revealed impairments in IL-10 production that correlated with lower levels of STAT3 phosphorylation, without abnormal expression of IL-10 receptor on Breg cells. On the other hand, BAFF levels were substantially elevated in SCD patients with SLE, but not significantly associated with Breg cell levels. Collectively, these results indicated numerical and functional deficits of Breg cells in SCD patients with SLE and their capacity to maintain tolerance and control inflammation is imbalanced, which leads to the development of autoimmune responses.


Evaluating oversight of human drugs and medical devices: a case study of the FDA and implications for nanobiotechnology.
This article evaluates the oversight of drugs and medical devices by the U.S. Food and Drug Administration (FDA) using an integration of public policy, law, and bioethics approaches and employing multiple assessment criteria, including economic, social, safety, and technological. Criteria assessment and expert elicitation are combined with existing literature, case law, and regulations in an integrative historical case studies approach. We then use our findings as a tool to explore possibilities for effective oversight and regulatory mechanisms for nanobiotechnology. Section I describes oversight mechanisms for human drugs and medical devices and presents current nanotechnology products. Section II describes the results of expert elicitation research. Section III highlights key criteria and relates them to the literature and larger debate. We conclude with broad lessons for the oversight of nanobiotechnology informed by Sections I-III in order to provide useful analysis from multiple disciplines and perspectives to guide discussions regarding appropriate FDA oversight.


Experimental studies of extinction dynamics
Extinction of populations occurs naturally, but global extinction rates are accelerating, making understanding extinction a high priority for conservation. Extinction in experimental populations of brine shrimp (Artemia franciscana) was measured to assess hypothesized extinction processes. Greater initial population size, greater maximum population size supported by the environment, and lower variation in environmental conditions reduced the likelihood of extinction, as hypothesized. However, initial population size was less important, and maximum population size and environmental variation were more important than often hypothesized. Unexpectedly, deterministic oscillations in population size due to inherent nonlinear dynamics and overcrowding were as important or more important than hypothesized processes.


Impact of organic load on Escherichia coli O157:H7 survival during pilot-scale processing of iceberg lettuce with acidified sodium hypochlorite.
Chemical sanitizers are routinely used during commercial flume washing of fresh-cut leafy greens to minimize cross-contamination from the water. This study assessed the efficacy of three chlorine treatments against Escherichia coli O157:H7 on iceberg lettuce, in wash water, and on surfaces of a pilot-scale processing line using flume water containing various organic loads. Iceberg lettuce (5.4 kg) was inoculated to contain 10(6) CFU/g of a 4-strain cocktail of nontoxigenic, green fluorescent protein-labeled, ampicillin-resistant E. coli O157:H7 and held for 24 h at 4°C before processing. Lettuce was shredded using a Urschel TransSlicer, step conveyed to a flume tank, washed for 90 s using water alone or one of three different sanitizing treatments (50 ppm of total chlorine either alone or acidified to pH 6.5 with citric acid or T-128) in water containing organic loads of 0, 2.5, 5, or 10% (wt/vol) blended iceberg lettuce, and then dried using a shaker table and centrifugal dryer. Next, three 5.4-kg batches of uninoculated iceberg lettuce were processed identically. Various product (25 g), water (50 ml), and equipment surface swab (100 cm(2)) samples were homogenized in neutralizing buffer, diluted appropriately, and plated on tryptic soy agar containing 0.6% (wt/vol) yeast extract and 100 ppm of ampicillin without prior 0.45- m m membrane filtration to quantify E. coli O157:H7. Organic load negatively impacted the efficacy of all three chlorine treatments (P < 0.05) at the end of processing, with typical E. coli O157:H7 reductions of >5 and 0.9 to 3.7 log CFU/ml for organic loads of 0 and 10%, respectively. Organic load rarely had a significant impact (P < 0.05) on the efficacy of chlorine, chlorine plus citric acid, or chlorine plus T-128 against E. coli O157:H7 on iceberg lettuce. Reduced sanitizer efficacy generally corresponded to changes in total solids, chemical oxygen demand, turbidity, and maximum filterable volume, indicating that these tests may be effective alternatives to the industry standard of oxygen/reduction potential.


Psychosocial stressors and low birthweight in an urban population.
Low birthweight is a major determinant of infant mortality, as well as a contributor to infant and childhood morbidity. A key issue is how to reduce the incidence of low birthweight in the United States. One emerging factor is exposure to psychosocial stressors. In this research, we evaluated the association between exposure to psychosocial stressors and low birthweight in a population of urban, low-income pregnant women. Over 2,000 pregnant women 18 years of age and older were enrolled in this prospective study and recruited at their first prenatal care visit. We obtained information on maternal exposure to stressors. After the pregnancy, we abstracted clinical records of each woman enrolled in the study. Logistic regression was used to estimate the adjusted odds ratio for the association between stressor group membership and low birthweight, controlling for the effects of confounding factors. In logistic regression analyses stratified by race, for African-American women, the following variables were significantly associated with low birthweight: smoking, hypertension, low prepregnancy weight, hospitalization during pregnancy, previous preterm birth, and exposure to stressors. For Caucasian women, significant predictors were: smoking, drug use, hospitalization during pregnancy, hypertension, and previous preterm birth. Exposure to stressors was also significantly associated with many clinical and behavioral risks for low birthweight. Our results suggest two potential mechanisms for an association between stressors and low birthweight. Exposure to stressors may be indirectly associated with low birthweight through a relationship with clinical and behavioral risks for low birthweight. Exposure to psychosocial stressors may also be directly associated with risk of low birthweight among African-American women.


ADAMTS13 content in plasma-derived factor VIII/von Willebrand factor concentrates.
Thrombotic thrombocytopenic purpura (TTP) is a microangiopathy syndrome caused by a congenital or acquired deficiency of ADAMTS13, a plasma metalloprotease that cleaves von Willebrand factor (VWF) and thus prevents the formation of platelet-rich thrombi in the microcirculation. TTP can be fatal if not appropriately and timely treated with the infusion of fresh frozen plasma (FFP) or exchange plasmapheresis, that reverse the process of microangiopathy by removing anti-ADAMTS13 autoantibodies and replacing functional ADAMTS13. The treatment of TTP with FFP is not free from risks and must be administered in hospitals or clinics, owing to the substantial amount of plasma volume infused or exchanged and the frequent need of catheter application. Moreover, most FFPs are not subjected to treatments to remove or inactivate blood-borne infectious agents. A number of recent reports indicate that certain plasma-derived VWF-factor VIII (FVIII) concentrates are clinically effective in the treatment of congenital TTP. In this study, we measured ADAMTS13 levels in various plasma-derived VWF-FVIII concentrates, showing that Koate(®) -DVI (Grifols), contained relatively high amounts of ADAMTS13 and that Alphanate(®) (Grifols) was the closest other product in terms of protease content. Koate(®) -DVI contains, on average (five lots tested), 0.091 ± 0.007 Units of ADAMTS13 activity per IU of FVIII. On the basis of this analysis and other reports of VWF-FVIII concentrate utilization in congenital TTP, potential dosing, and future clinical developments are discussed.


Acquiring contextualized concepts: a connectionist approach.
Conceptual knowledge is acquired through recurrent experiences, by extracting statistical regularities at different levels of granularity. At a fine level, patterns of feature co-occurrence are categorized into objects. At a coarser level, patterns of concept co-occurrence are categorized into contexts. We present and test CONCAT, a connectionist model that simultaneously learns to categorize objects and contexts. The model contains two hierarchically organized CALM modules (Murre, Phaf, & Wolters, 1992). The first module, the Object Module, forms object representations based on co-occurrences between features. These representations are used as input for the second module, the Context Module, which categorizes contexts based on object co-occurrences. Feedback connections from the Context Module to the Object Module send activation from the active context to those objects that frequently occur within this context. We demonstrate that context feedback contributes to the successful categorization of objects, especially when bottom-up feature information is degraded or ambiguous.


Framing risk in pandemic influenza policy and control.
This article explores differing understandings of 'risk' in relation to pandemic influenza policy and control. After a preliminary overview of methodological and practical problems in risk analysis, ways in which risk was framed and managed in three historical cases were examined. The interdependence between scientific empiricism and political decision-making led to the mismanagement of the 1976 swine influenza scare in the USA. The 2004 H5N1 avian influenza outbreak in Thailand, on the other hand, was undermined by questions of national economic interest and concerns over global health security. Finally, the recent global emergency of pandemic influenza H1N1 in 2009 demonstrated the difficulties of risk management under a context of pre-established perceptions about the characteristics and inevitability of a pandemic. Following the analysis of these cases, a conceptual framework is presented to illustrate ways in which changing relationships between risk assessment, risk perception and risk management can result in differing policy strategies.


Species-specific ecological niche modelling predicts different range contractions for Lutzomyia intermedia and a related vector of Leishmania braziliensis following climate change in South America.
Before 1996 the phlebotomine sand fly Lutzomyia neivai was usually treated as a synonym of the morphologically similar Lutzomyia intermedia, which has long been considered a vector of Leishmania braziliensis, the causative agent of much cutaneous leishmaniasis in South America. This report investigates the likely range changes of both sand fly species in response to a stabilisation climate change scenario (RCP4.5) and a high greenhouse gas emissions one (RCP8.5). Ecological niche modelling was used to identify areas of South America with climates currently suitable for each species, and then the future distributions of these climates were predicted based on climate change scenarios. Compared with the previous ecological niche model of L. intermedia (sensu lato) produced using the GARP algorithm in 2003, the current investigation modelled the two species separately, making use of verified presence records and additional records after 2001. Also, the new ensemble approach employed ecological niche modelling algorithms (including Maximum Entropy, Random Forests and Support Vector Machines) that have been widely adopted since 2003 and perform better than GARP, as well as using a more recent climate change model (HadGEM2) considered to have better performance at higher resolution than the earlier one (HadCM2). Lutzomyia intermedia was shown to be the more tropical of the two species, with its climatic niche defined by higher annual mean temperatures and lower temperature seasonality, in contrast to the more subtropical L. neivai. These different latitudinal ranges explain the two species' predicted responses to climate change by 2050, with L. intermedia mostly contracting its range (except perhaps in northeast Brazil) and L. neivai mostly shifting its range southwards in Brazil and Argentina. This contradicts the findings of the 2003 report, which predicted more range expansion. The different findings can be explained by the improved data sets and modelling methods. Our findings indicate that climate change will not always lead to range expansion of disease vectors such as sand flies. Ecological niche models should be species specific, carefully selected and combined in an ensemble approach.


Molecular mechanisms of vascular permeability in diabetic retinopathy.
Diabetes leads to a wide array of complications in humans, including kidney failure, vascular disease, peripheral nerve degeneration, and vision loss. Diabetic retinopathy causes blindness in more working-age people in the United States than any other disease and contributes greatly to blindness in the young and old as well. The increasing rate of diabetes occurring in our society can only bring about a further decrease in the visual health of this country unless new modalities are discovered to prevent and cure diabetic retinopathy. Breakdown of the blood-retinal barrier and the resultant vascular permeability remains one of the first observable alterations in diabetic retinopathy and strongly correlates with vision loss. In this article, we examine the molecular components that form this blood-retinal barrier and explore how changes in the production of growth factors in the neural parenchyma cause an increase in vascular permeability and contribute to retinal degeneration.


Identification of basal and cyclic AMP regulatory elements in the promoter of the phosphoenolpyruvate carboxykinase gene.
Promoter elements important for basal and cyclic AMP (cAMP)-regulated expression of the phosphoenolpyruvate carboxykinase (PEPCK) gene have been identified by analysis of a series of PEPCK promoter mutations in transfection experiments. Fusion genes containing wild-type and mutated PEPCK promoter sequences from -600 to +69 base pairs (bp) fused to the coding sequence for chloramphenicol acetyltransferase were studied. Internal deletion mutations that replaced specific bases with a 10-bp linker within the region from -129 bp to -18 bp of the PEPCK promoter were examined. In addition, wild-type and mutated DNA templates were used as probes in DNase I protection experiments to determine sites of protein-DNA interaction. The PEPCK promoter contains a binding site for nuclear factor 1-CAAT. Deletion of the 5' end of this binding site reduced the size of the DNase I footprint in this region but had no effect on promoter activity. In contrast, deletion or disruption of the 3' end of this binding site completely eliminated protein binding and reduced promoter activity by 50%. Deletion of core sequences of the cAMP regulatory element (CRE) resulted in loss of cAMP responsiveness and an 85% decrease in basal promoter activity, indicating that the CRE also functions as a basal stimulatory element. Mutation of the core sequence of the CRE resulted in loss of the DNase I footprint over the CRE. Internal deletions flanking the CRE showed no loss of induction by cAMP but did have reduced promoter activity. This delimits the CRE to an 18-bp region between nucleotides -100 and -82. Analysis of mutations that disrupted bases between the CRE and the initiation site identified a basal inhibitory element adjacent to a basal stimulatory element, both located just 3' of the CRE, as well as a basal stimulatory element coincident with the TATA consensus sequence centered at -27. These data demonstrate that several cis-acting elements are located within 130 nucleotides of the initiation site of the PEPCK gene and that the CRE is essential for both basal promoter activity and cAMP-regulated expression of this gene.


Healthy life skills and related factors among university students: a cross-sectional study in Istanbul, Turkey.
Chronic non-communicable diseases (NCDs) are the leading cause of global deaths. University students with unhealthy lifestyle constitute a high-risk group for NCDs. Evaluating and developing healthy behaviors during this period is very important for future health outcomes. This study was conducted to determine healthy life skills in university students. A cross-sectional study was conducted at a university, and data were collected between January and March 2023 in Istanbul. Data were collected using the healthy living skills scale in University Students and the Personal Information Form. Normal distribution conditions of the data were checked with the Kolmogorov-Smirnov test. According to data distribution, data with normal distribution were analyzed using parametric statistics including t-test, ANOVA and Chi-square tests and data with non-normal distribution were analyzed using non-parametric tests including Mann-Whitney U-test and Fisher's exact test. Logistic regression test was used to determine predictor variables. The average score for healthy life skills was 63.5 out of 84. Significant differences were found in the scores based on marital status, economic income, social security insurance coverage, and educational grade (p = 0.03, p = 0.001, p = 0.004, p = 0.04, respectively). Students who reported alcohol and smoking consumption had lower scores (60.8 ± 12) out of 84. The study revealed that university students in Istanbul possess a satisfactory level of healthy life skills. By providing social support, such as expanding the coverage of social security insurance and establishing conducive educational environments, while also paying attention to the influence of peers on students, we can contribute to the development of healthy life skills in university students.


Spinal manifestations of Ehlers-Danlos syndrome: a scoping review.
Since its initial description, the definition of Ehlers-Danlos syndrome (EDS) has notably changed. At present, it broadly refers to disorders of the connective tissue that are heritable and have similar features including joint hypermobility, dermal dysplasia, and vascular as well as internal organ fragility. There has been no comprehensive review of spinal manifestations of EDS in the recent literature. That has led to controversies in management protocols of this so-called orphan disease. The authors used the latest version of the EDS classification from 2017, in which 13 subtypes were recognized. EDS has 19 different causal genes, mainly associated with collagen synthesis. Of these, 5 subtypes have associated spinal manifestations. Some of the spinal pathologies associated with EDS include Chiari malformation, craniocervical instability, kyphoscoliosis, segmental instability and kyphosis, spontaneous CSF leaks, Tarlov cyst syndrome, tethered cord, and problems associated with wound healing. Here, the authors briefly discuss the demographics, etiology, pathophysiology, clinical features, management strategies, and directions for further research for each of these manifestations. EDS belongs to the group of orphan diseases, with the total patient population being below 200,000. Further research on spinal manifestations of EDS is the need of the hour to establish clinical practice guidelines and close the significant knowledge gaps that currently exist.