Daily rhythms of behavioral and hormonal patterns in male dromedary camels housed in boxes.
Daily rhythmicity has been observed for a number of hormonal and behavioral variables in mammals. It can be entrained by several external factors, such as light-dark cycle and scheduled feeding. In dromedary camels, daily rhythmicity has been documented only for melatonin secretion and body temperature. In this study, the daily rhythmicity of behavioral repertoire, cortisol and testosterone levels was investigated in captive male camels. Six clinically healthy male dromedary camels (Camelus dromedarius) were used. The animals were housed in single boxes for 24 h daily and fed twice a day. Over a period of 48 h, behavioral observations were made and blood samples taken every two hours. The data were analyzed using diurnality index, conisor analysis and PROC mixed procedure. The diurnality index for rumination and lying down was close to 0 (respectively, 0.09 and 0.19), while the indices for stereotypy, standing, feeding and walking were close to 1 (respectively, 0.74, 0.84, 0.92 and 0.85). Cosinor analysis revealed daily rhythmicity for all behaviors and for cortisol levels (acrophase at 12:57) but not for testosterone. Rumination and lying down (inactive behaviors) reached a peak during the scotophase, whereas feeding, walking and stereotypy (active behaviors) reached a peak during the photophase around midday. Cortisol level and expression of stereotypies peaked before and after food distribution and were negatively correlated (r = - 0.287, P = 0.005). Testosterone levels and expression of sexual behaviors were stimulated by the visual and olfactory contacts with the females and were positively correlated (r = 0.164, P = 0.040). Testosterone was also negatively correlated with cortisol (r = - 0.297; P = 0.003). These preliminary results provided new knowledge about the daily rhythm of behaviors in camels housed in boxes, suggesting that camels exhibit diurnal behavior pattern in the maintenance conditions outlined in the study. Daily rhythmicity seemed to be entrained not only by the light-dark cycle but also by scheduled feeding. The rise in stereotypy after food distribution could be due to the persistence of feeding motivation and frustration after the ingestion of food. Therefore, feeding practices should be improved to satisfy the foraging and feeding motivation of these camels. Behavioral and hormonal daily patterns in camels should be taken in consideration to adapt the management system, giving the animals more freedom during the light period and a diet richer in fiber, so as to improve reproductive performance, health and welfare.


To bead or not to bead? Lens-specific intermediate filaments revisited.
For nearly three decades cytoplasmic intermediate filaments (IFs) have been described as 10 nm thick, unbranched ropes radiating from the cell nucleus and extending to the plasma membrane. This stereotype is now being challenged by the discovery and molecular characterization of the beaded filaments (BFs), a novel class of IFs composed of the lens-specific proteins filensin and phakinin. In contrast to 'mainstream' IFs, BFs have a distinctly nodular appearance and form a meshwork underneath the plasma membrane of the lens fiber cells. In vitro assembly studies, expression of filensin and phakinin in cultured cells, and analysis of the corresponding genes reveal that these proteins have evolved from two different subfamilies of IF proteins, thus yielding a unique structure. The new information provides a basis for understanding how the various forms of tissue-specific IF proteins might have developed adopting to the constraints of a specialized environment.


Fast burst firing and short-term synaptic plasticity: a model of neocortical chattering neurons.
We present an ionic conductance model of chattering neurons in the neocortex, which fire fast rhythmic bursts in the gamma frequency range (approximately 40 Hz) in response to stimulation [Gray C. M. and McCormick D. A. (1996) Science 274, 109-113]. The bursting mechanism involves a "ping-pong" interplay between soma-to-dendrite back propagation of action potentials and an afterdepolarization generated by a persistent dendritic Na+ current and a somatic Na+ window current. The oscillation period is primarily determined by a slowly inactivating K+ channel and passive membrane properties. The model behavior is compared quantitatively with the experimental data. It is shown that the cholinergic muscarinic receptor activation can transform the model cell's firing pattern from tonic spiking to rapid bursting, as a possible pathway for acetylcholine to promote 40-Hz oscillations in the visual cortex. To explore possible functions of fast burst firing in the neocortex, a hypothetical neural pair is simulated, where a chattering cell is presynaptic to an inhibitory interneuron via stochastic synapses. For this purpose, we use a synapse model endowed with a low release probability, short-term facilitation and vesicle depletion. This synapse model reproduces the behavior of certain neocortical pyramid-to-interneuron synapses [Thomson A. M. et al. (1993) Neuroscience 54, 347-360]. We showed that the burstiness of cell firing is required for the rhythmicity to be reliably transmitted to the postsynaptic cell via unreliable synapses, and that fast burst firing of chattering neurons can provide an exceptionally powerful drive for recruiting feedback inhibition in cortical circuits. From these results, we propose that the fast rhythmic burst firing of neocortical chattering neurons is generated by a calcium-independent ionic mechanism. Our simulation results on the neural pair highlight the importance of characterizing the short-term plasticity of the synaptic connections made by chattering cells, in order to understand their putative pacemaker role in synchronized gamma oscillations of the visual cortex.


Plasma Metabolite Markers of Parkinson's Disease and Atypical Parkinsonism.
Differentiating between Parkinson's disease (PD) and the atypical Parkinsonian disorders of multiple system atrophy (MSA) and progressive supranuclear palsy (PSP) is difficult clinically due to overlapping symptomatology, especially at early disease stages. Consequently, there is a need to identify metabolic markers for these diseases and to develop them into viable biomarkers. In the present investigation, solution nuclear magnetic resonance and mass spectrometry metabolomics were used to quantitatively characterize the plasma metabolomes (a total of 167 metabolites) of a cohort of 94 individuals comprising 34 PD, 12 MSA, and 17 PSP patients, as well as 31 control subjects. The distinct and statistically significant differences observed in the metabolite concentrations of the different disease and control groups enabled the identification of potential plasma metabolite markers of each disorder and enabled the differentiation between the disorders. These group-specific differences further implicate disturbances in specific metabolic pathways. The two metabolites, formic acid and succinate, were altered similarly in all three disease groups when compared to the control group, where a reduced level of formic acid suggested an effect on pyruvate metabolism, methane metabolism, and/or the kynurenine pathway, and an increased succinate level suggested an effect on the citric acid cycle and mitochondrial dysfunction.


The Chemokine Platelet Factor-4 Variant (PF-4var)/CXCL4L1 Inhibits Diabetes-Induced Blood-Retinal Barrier Breakdown.
To investigate the expression of platelet factor-4 variant (PF-4var/CXCL4L1) in epiretinal membranes from patients with proliferative diabetic retinopathy (PDR) and the role of PF-4var/CXCL4L1 in the regulation of blood-retinal barrier (BRB) breakdown in diabetic rat retinas and human retinal microvascular endothelial cells (HRMEC). Rats were treated intravitreally with PF-4var/CXCL4L1 or the anti-vascular endothelial growth factor (VEGF) agent bevacizumab on the first day after diabetes induction. Blood-retinal barrier breakdown was assessed in vivo with fluorescein isothiocyanate (FITC)-conjugated dextran and in vitro in HRMEC by transendothelial electrical resistance and FITC-conjugated dextran cell permeability assay. Occludin, vascular endothelial (VE)-cadherin, hypoxia-inducible factor (HIF)-1α, VEGF, tumor necrosis factor (TNF)-α, receptor for advanced glycation end products (RAGE), caspase-3 levels, and generation of reactive oxygen species (ROS) were assessed by Western blot, enzyme-linked immunosorbent assays, or spectrophotometry. In epiretinal membranes, vascular endothelial cells and stromal cells expressed PF-4var/CXCL4L1. In vitro, HRMEC produced PF-4var/CXCL4L1 after stimulation with a combination of interleukin (IL)-1β and TNF-α, and PF-4var/CXCL4L1 inhibited VEGF-mediated hyperpermeability in HRMEC. In rats, PF-4var/CXCL4L1 was as potent as bevacizumab in attenuating diabetes-induced BRB breakdown. This effect was associated with upregulation of occludin and VE-cadherin and downregulation of HIF-1α, VEGF, TNF-α, RAGE, and caspase-3, whereas ROS generation was not altered. Our findings suggest that increasing the intraocular PF-4var/CXCL4L1 levels early after the onset of diabetes protects against diabetes-induced BRB breakdown.


An ultrastructural evaluation of the cell organelle specificity of the uranaffin reaction in two human endocrine neoplasms.
The organelle specificity of the uranaffin reaction was determined by subjecting two human neoplasms (a pheochromocytoma and islet cell carcinoma) to four different experimental conditions. In Uranaffin Procedure (UP) I, fixed tissue was immersed in 0.9% sodium chloride (NaCl) before reacting with 4% uranyl acetate (pH 3.9) for 24 hours. In UP II, the tissue was prepared as in UP I with the exception that the tissue was immersed in uranyl acetate for 48 hours. In UP III, fixed tissue was prepared as in UP I with the exception that tissue was immersed in 0.1M cacodylate buffer (pH 7.2) instead of 0.9% NaCl. In UP IV, fixed tissue was prepared as in UP III with the exception that the tissue was immersed in uranyl acetate for 48 hours instead of 24 hours. When UPs I and II were utilized, only three cell organelles showed electron-dense reactivity: the nucleus, ribosomes, and cytoplasmic neurosecretory-like granules. In the nucleus, the nuclear chromatin, nucleolus, interchromatinic granules and perichromatinic granules were intensely stained. The reaction product in all of the uranaffin-positive organelles had a finely granular appearance. When fixed tissue was immersed in cacodylate buffer instead of isotonic saline, a non-specific reactivity was observed. The reaction product in some areas had a distinct crystalline appearance and filled some areas of the cytosol, the cisternae of the endoplasmic reticulum, the Golgi apparatus, the perinuclear cisternae, the nucleus, nucleolus, mitochondria, neurosecretory-like granules and larger lysosome-like bodies. There was a statistically significant (p less than 0.05) increase in the number of uranaffin-positive granules/mu2 when both endocrine neoplasms were reacted with uranyl acetate for 48 hours instead of 24 hours. The increase in uranaffin-positive granules using UP II did not result in an increase in non-specificity of the staining reaction.


Inflammatory cytokines and a diverse cervicovaginal microbiota associate with cervical dysplasia in a cohort of Hispanics living in Puerto Rico.
Cervical cancer (CC) is women's fourth most common cancer worldwide. A worrying increase in CC rates in Hispanics suggests that besides Human papillomavirus infections, there may be other cofactors included in the epithelial microenvironment that could play a role in promoting the disease. We hypothesized that the cervical microbiome and the epithelial microenvironment favoring inflammation is conducive to disease progression in a group of Hispanics attending gynecology clinics in Puerto Rico. Few studies have focused on the joint microbiota and cytokine profile response in Hispanics outside the US, especially regarding the development of precancerous lesions. We aimed to investigate the relationship between the cervicovaginal microbiome and inflammation in Hispanic women living in PR while considering cervical dysplasia and HPV genotype risk. Cervical samples collected from 91 participants coming to gynecology clinics in San Juan, underwent 16S rRNA genes (V4 region) profiling, and cytokines were measured using Luminex MAGPIX technology. Cytokines were grouped as inflammatory (IL-1β, TNFα, IFNγ, IL-6), anti-inflammatory (IL- 4, IL-10, TGFβ1), and traffic-associated (IL-8, MIP1a, MCP1, IP10). They were related to microbes via an inflammation scoring index based on the quartile and tercile distribution of the cytokine's concentration. We found significant differences in the diversity and composition of the microbiota according to HPV type according to carcinogenic risk, cervical disease, and cytokine abundance. Community State Types (CSTs) represents a profile of microbial communities observed within the vaginal microbiome ecological niche, and Lactobacillus-depleted CST IV had ~ 90% dominance in participants with high-grade squamous intraepithelial lesions and high-risk HPV. The increasing concentration of pro-inflammatory cytokines was associated with a decrease in L. crispatus. In contrast, dysbiosis-associated bacteria such as Gardnerella, Prevotella, Atopobium concomitantly increased with pro-inflammatory cytokines. Our study highlights that the cervical microbiota of Hispanics living in Puerto Rico is composed mostly of diverse CST profiles with decreased Lactobacillus and is associated with a higher pro-inflammatory environment. The joint host-microbe interaction analyses via cytokine and microbiota profiling have very good translational potential.


Oral infection with Porphyromonas gingivalis induces peri-implantitis in a murine model: Evaluation of bone loss and the local inflammatory response.
Peri-implantitis is a major health concern, with unclear pathogenesis, and with no accessible animal models. Our aim was to establish a mouse model for peri-implantitis and to investigate mediators of inflammation. Mice were divided into implanted versus non-implanted groups. Implants were inserted immediately following the extraction of the upper first molar. Four weeks following implantation, implanted and non-implanted mice were challenged with either Porphyromonas gingivalis or vehicle (eight mice in each subgroup, 32 mice in total). Alveolar bone loss and expression of inflammatory mediators in the soft tissue were assessed 42 days following infection. Porphyromonas gingivalis infection induced greater bone loss around implants than around teeth. In non-infected animals, the presence of the implant correlated with elevated expression of Il-10, Foxp3 and Rankl/Opg ratio, while Tnf-α levels were decreased relative to tissue around teeth. Six weeks following infection, Tnf-α increased significantly while the expression of Foxp3 decreased in the tissue around the implants. No significant differences in anti- or pro-inflammatory mediators were found around teeth of infected, relative to non-infected mice. Oral infection with P. gingivalis of mice with implants induced bone loss and a shift in gingival cytokine expression. This mouse model enables exploration of the pathogenesis of peri-implantitis and testing of novel treatments.


Increased ethanol accumulation from glucose via reduction of ATP level in a recombinant strain of Saccharomyces cerevisiae overexpressing alkaline phosphatase.
The production of ethyl alcohol by fermentation represents the largest scale application of Saccharomyces cerevisiae in industrial biotechnology. Increased worldwide demand for fuel bioethanol is anticipated over the next decade and will exceed 200 billion liters from further expansions. Our working hypothesis was that the drop in ATP level in S. cerevisiae cells during alcoholic fermentation should lead to an increase in ethanol production (yield and productivity) with a greater amount of the utilized glucose converted to ethanol. Our approach to achieve this goal is to decrease the intracellular ATP level via increasing the unspecific alkaline phosphatase activity. Intact and truncated versions of the S. cerevisiae PHO8 gene coding for vacuolar or cytosolic forms of alkaline phosphatase were fused with the alcohol dehydrogenase gene (ADH1) promoter. The constructed expression cassettes used for transformation vectors also contained the dominant selective marker kanMX4 and S. cerevisiae δ-sequence to facilitate multicopy integration to the genome. Laboratory and industrial ethanol producing strains BY4742 and AS400 overexpressing vacuolar form of alkaline phosphatase were characterized by a slightly lowered intracellular ATP level and biomass accumulation and by an increase in ethanol productivity (13% and 7%) when compared to the parental strains. The strains expressing truncated cytosolic form of alkaline phosphatase showed a prolonged lag-phase, reduced biomass accumulation and a strong defect in ethanol production. Overexpression of vacuolar alkaline phosphatase leads to an increased ethanol yield in S. cerevisiae.


Effect of clearcutting operations on the survival rate of a small mammal.
Clearcutting is a common timber harvesting technique that represents a significant and abrupt change in habitat conditions for wildlife living in industrial forests. Most research on this type of impact has focused on comparing populations or communities in mature forests/plantations and the resulting clearcut stands. However, this approach does not separate the effect of changes in habitat attributes from direct mortality produced by the intensive use of heavy machinery required for cutting down trees and dragging them to a road. Because knowing the fate of individuals after a disturbance is important for modelling landscape-scale population dynamics in industrial forests, we conducted a study in South-Central Chile to understand the short-term response to clearcutting operations of the long-haired Akodont (Abrothrix longipillis), a forest specialist mouse. Between 2009 and 2013 we radiotracked a total of 51 adult male Akodonts, before, during and after the clearcutting of the pine plantations in which they lived. A minimum of 52.4% of the individuals died as a direct cause of the timbering operations, being crushed by vehicles or logs during logging operations. Our observations suggest that, instead of fleeing the area, the response of long-haired Akodonts to the approaching machinery is to hide under the forest litter or in burrows, which exposes them to a serious risk of death. The real mortality rate associated to clearcutting may be higher than that estimated by us because of some methodological biases (i.e. individuals with crushed radiotransmitters not recorded) and the fact that additional mortality sources may affect the population in the weeks following logging operations (e.g., higher exposure to predation, effects of site preparation for the new plantation, etc).


Population-based incidence and outcome of acetaminophen poisoning by type of ingestion.
1) To determine, in a population-based sample, the observed frequency of acetaminophen overdose-related ED evaluation and hospitalization. 2) To examine the relative frequency of hospitalization by pattern of ingestion, the outcome of each group, and the presence or absence of postulated risk factors. This study was a 46-month, retrospective chart review of all acetaminophen-related visits, by patients at least 10 years of age, to either of the two hospitals that serve a four-county region of central Virginia. Of 636 charts identified for review, only 137 involved acute or chronic acetaminophen overdose. One hundred twenty-six patients presented after an acute ingestion; 122 of these patients gave a history of a single, supratherapeutic ingestion of acetaminophen. Twenty-five patients were hospitalized for treatment. Eighteen of these were treated with N-acetylcysteine (NAC) based on the Rumack-Matthew nomogram; one suffered significant hepatic injury. The other seven presented at least 18 hours after ingestion, with no measurable serum acetaminophen. Two of these suffered significant hepatic injury. Four additional patients presented after multiple ingestions within 24 hours. Three were hospitalized, but none experienced significant injury. Only 11 patients were evaluated for chronic acetaminophen overmedication for pain (more than 6 g/day over a period of more than 24 hours). Four were admitted for treatment; three suffered significant hepatic injury. Thus, the observed incidence of acute acetaminophen ingestion in this region was 21.4/100,000/yr (95% CI = 17.7 to 25.2). The observed incidence of hospitalization for acute acetaminophen toxicity was 4.8/100,000/yr (95% CI = 3.0 to 6.5). The observed incidence of hospitalization for all acetaminophen poisoning was 5.5/100,000/yr (95% CI = 4.1 to 7.0). High ethanol consumption was present more frequently in those who suffered hepatic injury. Most patients evaluated for acetaminophen ingestion present early following acute single overdose. Relatively few of these patients require hospitalization and, for those hospitalized, the outcome is good. More significantly, acetaminophen overdose patients whose risk cannot be estimated using the Rumack-Matthew nomogram represented 44% of those hospitalized and 83% of those who suffered significant hepatic injury. Emergency physicians need to determine how they can impact the outcome of these patients. Efforts should be directed at further characterizing historical, physical, and biochemical markers of risk and at determining in which circumstances hospitalization for NAC or other therapies is justified.


Liability and managed care. American Academy of Pediatrics, Committee on Medical Liability.
This statement is intended to inform practitioners of the liability issues arising from managed care arrangements. Although it is not possible for pediatricians to completely insulate themselves from all liability in these areas, this statement offers a number of strategies to decrease the chances of being successfully sued. However, because case law within this realm is constantly evolving in each state, these serve only as guidelines and are subject to both local and emerging developments.


Neuroinflammation and microglia: considerations and approaches for neurotoxicity assessment.
The impact of an inflammatory response, as well as interactions between the immune and nervous systems, are rapidly assuming major roles in neurodegenerative disease and injury. However, it is now appreciated that the exact nature of such responses can differ with each type of insult and interaction. More recently, neuroinflammation and the associated cellular response of microglia are being considered for their contribution to neurotoxicity of environmental agents; yet, so far, the inclusion of inflammatory end points into neurotoxicity assessment have relied primarily on relatively limited measures or driven by in vitro models of neurotoxicity. To present background information on relevant biological considerations of neuroinflammation and the microglia response demonstrating the complex integrative nature of these biological processes and raising concern with regards to translation of effects demonstrated in vitro to the in vivo situation. Specific points are addressed that would influence the design and interpretation of neuroinflammation with regards to neurotoxicology assessment. There is a complex and dynamic response in the brain to regulate inflammatory processes and maintain a normal homeostatic level. The classification of such responses as beneficial or detrimental is an oversimplification. Neuroinflammation should be considered as a balanced network of processes in which subtle modifications can shift the cells toward disparate outcomes. The tendency to overinterpret data obtained in an isolated culture system should be discouraged. Rather, the use of cross-disciplinary approaches to evaluate several end points should be incorporated into the assessment of inflammatory contributions to the neurotoxicity of environmental exposures.


Influence of year-on-year performance on final degree classification in a chiropractic master's degree program.
We explored if any predictors of success could be identified from end-of-year grades in a chiropractic master's program and whether these grades could predict final-year grade performance and year-on-year performance. End-of-year average grades and module grades for a single cohort of students covering all academic results for years 1-4 of the 2013 graduating class were used for this analysis. Analysis consisted of within-year correlations of module grades with end-of-year average grades, linear regression models for continuous data, and logistic regression models for predicting final degree classifications. In year 1, 140 students were enrolled; 85.7% of students completed the program 4 years later. End-of-year average grades for years 1-3 were correlated (Pearson r values ranging from .75 to .87), but the end-of-year grades for years 1-3 were poorly correlated with clinic internship performance. In linear regression, several modules were predictive of end-of-year average grades for each year. For year 1, logistic regression showed that the modules Physiology and Pharmacology and Investigative Imaging were predictive of year 1 performance (odds ratio [OR] = 1.15 and 0.9, respectively). In year 3, the modules Anatomy and Histopathology 3 and Problem Solving were predictors of the difference between a pass/merit or distinction final degree classification (OR = 1.06 and 1.12, respectively). Early academic performance is weakly correlated with final-year clinic internship performance. The modules of Anatomy and Histopathology year 3 and Problem Solving year 3 emerged more consistently than other modules as being associated with final-year classifications.


High resolution electroencephalography in freely moving mice.
Electroencephalography (EEG) is a standard tool for monitoring brain states in humans. Understanding the molecular and cellular mechanisms underlying diverse EEG rhythms can be facilitated by using mouse models under molecular, pharmacological, or electrophysiological manipulations. The small size of the mouse brain, however, poses a severe limitation in the spatial information of EEG. To overcome this limitation, we devised a polyimide based microelectrode array (PBM array) with nanofabrication technologies. The microelectrode contains 32 electrodes, weighs 150 mg, and yields noise-insensitive signals when applied on the mouse skull. The high-density microelectrode allowed both global and focused mapping of high resolution EEG (HR-EEG) in the mouse brain. Mapping and dynamical analysis tools also have been developed to visualize the dynamical changes of spatially resolved mouse EEG. We demonstrated the validity and utility of mouse EEG in localization of the seizure onset in absence seizure model and phase dynamics of abnormal theta rhythm in transgenic mice. Dynamic tracking of the EEG map in genetically modified mice under freely moving conditions should allow study of the molecular and cellular mechanisms underlying the generation and dynamics of diverse EEG rhythms.


Injury patterns in big ten conference football.
In 1998, the National Collegiate Athletic Association legislated a decrease in the number of scrimmages and full-contact practices allowed during the spring season. A significantly increased risk of injury faced by the same player during the spring versus fall practice sessions does exist, but the National Collegiate Athletic Association regulations will not have the intended effect of reducing injury rates to equal or below those of the fall practice sessions. Retrospectively analyzed, descriptive study of prospectively collected injury surveillance data followed by a prospective, controlled, repeated-measures study after the rule change. Phase 1: the Big Ten Conference Sports Injury Surveillance System database was reviewed from spring 1992 through fall 1997 for all reportable injuries. Phase 2: a prospective investigation was instituted from spring 1998 through fall 2000. This protocol was modified to also document updated individual player position descriptions and string as well as exposures for the old fall and spring practice categories (scrimmages, full contact, and limited contact) and the new spring practice designations (full pads with and without tackling, helmets only, spring game, and other scrimmages). Phase 1: There were 3950 fall injuries and 1007 spring injuries, with 469 of the 1007 attributable to the "spring risk factor." There was a statistically significant increase in injury rate in the spring (19.8) versus fall (10.6). Scrimmages (incidence density ratio = 2.4) and limited-contact practices (incidence density ratio = 2.5) showed more than twice the fall injury rate. Phase 2: There were 1502 fall injuries and 648 spring injuries, with 192 attributable to the spring risk factor. There was a 3-fold increase in injury rate in the spring (incidence density ratio = 3.2). Although the noncontact, helmets-only practices produced the lowest injury rates, the nonspring game scrimmages and the limited-contact practices defined as "practice with pads but without tackling" displayed the highest injury rates. In both spring and fall, nonplayers exhibited the highest injury rates for all string groups. There was and still is a statistically significant increase in spring practice injury risk. The 1998 rule change resulted in an even greater increase in spring practice injury risk. If the goal is to minimize the number of spring practice injuries, it will be best accomplished by limiting the number of scrimmages and limited-contact sessions.


β-Galactosidase instructed supramolecular hydrogelation for selective identification and removal of senescent cells.
The identification and removal of senescent cells is very important to improve human health and prolong life. In this study, we introduced a novel strategy of β-galactosidase (β-Gal) instructed peptide self-assembly to selectively form nanofibers and hydrogels in senescent cells. We demonstrated that the in situ formed nanofibers could alleviate endothelial cell senescence by reducing p53, p21, and p16INK4a expression levels. We also demonstrated that our strategy could selectively remove senescent endothelial cells by inducing cell apoptosis, with an increase in the BAX/BCL-2 ratio and caspase-3 expression. Our study reports the first example of enzyme-instructed self-assembly (EISA) by a sugar hydrolase, which may lead to the development of supramolecular nanomaterials for the diagnosis and treatment of many diseases, such as cancer, and for other applications, such as wound healing and senescence.


Guarding the central venous access device: a new solution for an old problem.
CLABSIs are a major concern in both the adult and pediatric patient population. Contamination of catheter hubs is a common cause of CLABSI. A novel, transparent line guard protects CVAD hubs from gross contamination. Central line-associated blood stream infections (CLABSIs) are a serious and potentially deadly complication in patients with a central venous access device (CVAD). CVADs play an essential role in modern medicine, serving as lifelines for many patients. To maintain safe and stable venous access, infection prevention bundles are used to help protect patients from complications such as CLABSI. Despite most CLABSIs being preventable, rates have been on the rise, often disproportionately impacting critically ill children. New solutions are needed to strengthen infection prevention bundles and protect CVADs from pathogen entry at catheter hubs and line connections. A novel, Food and Drug Administration-listed device has become available recently to guard CVADs from sources of gross contamination, addressing this apparent gap in infection prevention technology and practice.


Monitoring of free-ranging and captive Psittacula populations in Western Europe for avian bornaviruses, circoviruses and polyomaviruses.
Avian pathogens such as bornaviruses, circoviruses and polyomaviruses are widely distributed in captive collections of psittacine birds worldwide and can cause fatal diseases. In contrast, only little is known about their presence in free-ranging psittacines and their impact on these populations. Rose-ringed parakeets (Psittacula krameri) and Alexandrine parakeets (Psittacula eupatria) are non-native to Europe, but have established stable populations in parts of Western Europe. From 2012-2017, we surveyed free-ranging populations in Germany and France as well as captive Psittacula individuals from Germany and Spain for avian bornavirus, circovirus and polyomavirus infections. Samples from two out of 469 tested free-ranging birds (0.4%; 95% confidence interval [CI-95]: 0.1-1.5%) were positive for beak and feather disease virus (BeFDV), whereas avian bornaviruses and polyomaviruses were not detected in the free-ranging populations. In contrast, avian bornaviruses and polyomaviruses, but not circoviruses were detected in captive populations. Parrot bornavirus 4 (PaBV-4) infection was detected by RT-PCR in four out of 210 captive parakeets (1.9%; CI-95: 0.7-4.8%) from four different holdings in Germany and Spain and confirmed by detection of bornavirus-reactive antibodies in two of these birds. Three out of 160 tested birds (1.9%; CI-95: 0.5-5.4%) possessed serum antibodies directed against budgerigar fledgling disease virus (BuFDV). PaBV-4 and BuFDV were also detected in several psittacines of a mixed holding in Germany, which had been in contact with free-ranging parakeets. Our results demonstrate that Psittacula parakeets are susceptible to common psittacine pathogens and their populations in Western Europe are exposed to these viruses. Nevertheless, the prevalence of avian bornaviruses, circoviruses and polyomaviruses in those populations is very low.RESEARCH HIGHLIGHTS Psittacula parakeets are susceptible to bornavirus, circovirus and polyomavirus infection.Introduced Psittacula populations in Europe have been exposed to these viruses.Nevertheless, they may be absent or present at only low levels in free-ranging Psittacula populations.Free-ranging populations in Europe pose a minor threat of transmitting these viruses to captive Psittaciformes.


In artificial roost comparison, bats show preference for rocket box style.
Understanding microhabitat preferences of animals is critical for effective conservation, especially for temperate-zone bats, which receive fitness benefits from selecting optimal roost microhabitats. Artificial roost structures are increasingly being used in conservation efforts for at-risk bat species. To evaluate microhabitat differences in common artificial roost structures and determine if roost selection occurs based on structure type, we installed artificial roosts of three different styles (bat box, rocket box, and bark mimic) in six clusters. We compared size and measured temperature parameters (12 points/roost) while bats were excluded from one cluster. We simultaneously conducted census counts during the active season at five more clusters open to bats for 1-2 years. The rocket box style provided larger entrance area, surface area, and volume versus other roost types. Microclimate varied with roost design. More positions inside the bat box and rocket box stayed within critical temperature limits for bats (0-45°C)-i.e., were usable. The bark-mimic provided less usable space than the rocket box and, often, large proportions of the roost were > 45°C. The rocket box provided the widest temperature availability in a given hour (max range available 7°C) and was more stable than the bark mimic. A maternity colony of Indiana bats (Myotis sodalis) selected the rocket box style; four of five available rocket boxes became primary maternity roosts, with 2-210 bats emerging per night. Future work should aim to manipulate roost size, temperature availability, and temperature stability in isolation to identify which features drive roost microhabitat selection by bats. Comparative studies of artificial roosts account for some inherent irregularity in natural systems, allowing us to study the dynamics of roost microhabitats. We recommend season-long monitoring of microhabitat in novel artificial refuges and comparative studies of artificial and natural roosts, and urge managers to consider potential positive and negative effects when substituting artificial roosts for natural habitat.


A first insight on the population structure of Mycobacterium tuberculosis complex as studied by spoligotyping and MIRU-VNTRs in Santiago, Chile.
Tuberculosis (TB) remains a significant public health problem worldwide, but the ecology of the prevalent mycobacterial strains, and their transmission, can vary depending on country and region. Chile is a country with low incidence of TB, that has a geographically isolated location in relation to the rest of South American countries due to the Andes Mountains, but recent migration from neighboring countries has changed this situation. We aimed to assess the genotypic diversity of Mycobacterium tuberculosis complex (MTBC) strains in Santiago, Chile, and compare with reports from other Latin-American countries. We analyzed MTBC isolates from pulmonary tuberculosis cases collected between years 2008 and 2013 in Central Santiago, using two genotyping methods: spoligotyping and 12-loci mycobacterial interspersed repetitive unit-variable number of tandem repeats (MIRU-VNTRs). Data obtained were analyzed and compared to the SITVIT2 database. Mean age of the patients was 47.5 years and 61% were male; 11.6% were migrants. Of 103 strains (1 isolate/patient) included, there were 56 distinct spoligotype patterns. Of these, 16 strains (15.5%) corresponded to orphan strains in the SITVIT2 database, not previously reported. Latin American and Mediterranean (LAM) (34%) and T (33%) lineages were the most prevalent strains, followed by Haarlem lineage (16.5%). Beijing family was scarcely represented with only two cases (1.9%), one of them isolated from a Peruvian migrant. The most frequent clustered spoligotypes were SIT33/LAM3 (10.7%), SIT53/T1 (8.7%), SIT50/H3 (7.8%), and SIT37/T3 (6.8%). We conclude that LAM and T genotypes are the most prevalent genotypes of MTBC in Santiago, Chile, and together correspond to almost two thirds of analyzed strains, which is similar to strain distribution reported from other countries of Latin America. Nevertheless, the high proportion of SIT37/T3, which was rarely found in other Latin American countries, may underline a specific history or demographics of Chile related to probable human migrations and evolutions.


Cognitive-behavioral therapy versus education and desipramine versus placebo for moderate to severe functional bowel disorders.
Studies of antidepressants and psychological treatments in functional bowel disorders (FBD) are methodologically limited. The aim of this study was to assess the clinical efficacy and safety of cognitive-behavioral therapy (CBT) against education (EDU) and desipramine (DES) against placebo (PLA) in female patients with moderate to severe FBD (irritable bowel syndrome, functional abdominal pain, painful constipation, and unspecified FBD). We also evaluated the amenability of clinically meaningful subgroups to these treatments. This randomized, comparator-controlled, multicenter trial enrolled 431 adults from the University of North Carolina and the University of Toronto with moderate to severe symptoms of FBD. Participants received psychological (CBT vs. EDU) or antidepressant (DES vs. PLA) treatment for 12 weeks. Clinical, physiologic, and psychosocial assessments were performed before and at the end of treatment. The intention-to-treat analysis showed CBT as significantly more effective than EDU (P = 0.0001; responder rate, 70% CBT vs. 37% EDU; number needed to treat [NNT ], 3.1). DES did not show significant benefit over PLA in the intention-to-treat analysis (P = 0.16; responder rate, 60% DES vs. 47% PLA; NNT, 8.1) but did show a statistically significant benefit in the per-protocol analysis (P = 0.01; responder rate, 73% DES vs. 49% PLA; NNT, 5.2), especially when participants with nondetectable blood levels of DES were excluded (P = 0.002). Improvement was best gauged by satisfaction with treatment. Subgroup analyses showed that DES was beneficial over PLA for moderate more than severe symptoms, abuse history, no depression, and diarrhea-predominant symptoms; CBT was beneficial over EDU for all subgroups except for depression. For female patients with moderate to severe FBD, CBT is effective and DES may be effective when taken adequately. Certain clinical subgroups are more or less amenable to these treatments.


Quantitative phospho-proteomics to investigate the polo-like kinase 1-dependent phospho-proteome.
Polo-like kinase 1 (PLK1) is a key regulator of mitotic progression and cell division, and small molecule inhibitors of PLK1 are undergoing clinical trials to evaluate their utility in cancer therapy. Despite this importance, current knowledge about the identity of PLK1 substrates is limited. Here we present the results of a proteome-wide analysis of PLK1-regulated phosphorylation sites in mitotic human cells. We compared phosphorylation sites in HeLa cells that were or were not treated with the PLK1-inhibitor BI 4834, by labeling peptides via methyl esterification, fractionation of peptides by strong cation exchange chromatography, and phosphopeptide enrichment via immobilized metal affinity chromatography. Analysis by quantitative mass spectrometry identified 4070 unique mitotic phosphorylation sites on 2069 proteins. Of these, 401 proteins contained one or multiple phosphorylation sites whose abundance was decreased by PLK1 inhibition. These include proteins implicated in PLK1-regulated processes such as DNA damage, mitotic spindle formation, spindle assembly checkpoint signaling, and chromosome segregation, but also numerous proteins that were not suspected to be regulated by PLK1. Analysis of amino acid sequence motifs among phosphorylation sites down-regulated under PLK1 inhibition in this data set identified two potential novel variants of the PLK1 consensus motif.


Usual nutrient intakes of US infants and toddlers generally meet or exceed Dietary Reference Intakes: findings from NHANES 2009-2012.
To our knowledge, few studies have described the usual nutrient intakes of US children aged <2 y or assessed the nutrient adequacy of their diets relative to the recommended Dietary Reference Intakes (DRIs). We estimated the usual nutrient intake of US children aged 6-23 mo examined in NHANES 2009-2012 and compared them to age-specific DRIs as applicable. Dietary intake was assessed with two 24-h recalls for infants aged 6-11 mo (n = 381) and toddlers aged 12-23 mo (n = 516) with the use of the USDA's Automated Multiple-Pass Method. Estimates of usual nutrient intakes from food and beverages were obtained with the use of the National Cancer Institute method. The proportions of children with intakes below and above the DRI were also estimated. The estimated usual intakes of infants were adequate for most nutrients; however, 10% had an iron intake below the Estimated Average Requirement (EAR), and only 21% had a vitamin D intake that met or exceeded the recommended Adequate Intake (AI). More nutrient inadequacies were noted among toddlers; 1 in 4 had a lower-than-recommended fat intake (percentage of energy), and most had intakes that were below the EAR for vitamins E (82%) and D (74%). Few toddlers (<1%) met or exceeded the AI for fiber and potassium. In contrast, 1 in 2 had sodium intakes that exceeded the Tolerable Upper Intake Level (UL); ≥16% and 41% of the children had excessive intakes (greater than the ULs) of vitamin A and zinc, respectively. The estimated usual intakes of infants were adequate for most nutrients. Most toddlers were at risk for inadequate intakes of vitamins D and E and had diets low in fiber and potassium. The sources contributing to excessive intakes of vitamin A and zinc among infants and toddlers may need further evaluation.


Progress Report on EMBED: A Pragmatic Trial of User-Centered Clinical Decision Support to Implement EMergency Department-Initiated BuprenorphinE for Opioid Use Disorder.
Buprenorphine (BUP) can safely and effectively reduce craving, overdose, and mortality rates in people with opioid use disorder (OUD). However, adoption of ED-initiation of BUP has been slow partly due to physician perception this practice is too complex and disruptive. We report progress of the ongoing EMBED (EMergency department-initiated BuprenorphinE for opioid use Disorder) project. This project is a five-year collaboration across five healthcare systems with the goal to develop, integrate, study, and disseminate user-centered Clinical Decision Support (CDS) to promote the adoption of Emergency Department (ED)-initiation of buprenorphine/naloxone (BUP) into routine emergency care. Soon to enter its third year, the project has already completed multiple milestones to achieve its goals including (1) user-centered design of the CDS prototype, (2) integration of the CDS into an automated electronic health record (EHR) workflow, (3) data coordination including derivation and validation of an EHR-based computable phenotype, (4) meeting all ethical and regulatory requirements to achieve a waiver of informed consent, (5) pilot testing of the intervention at a single site, and (6) launching a parallel group-randomized 18-month pragmatic trial in 20 EDs across 5 healthcare systems. Pilot testing of the intervention in a single ED was associated with increased rates of ED-initiated BUP and naloxone prescribing and a doubling of the number of unique physicians adopting the practice. The ongoing multi-center pragmatic trial will assess the intervention's effectiveness, scalability, and generalizability with a goal to shift the emergency care paradigm for OUD towards early identification and treatment. Clinicaltrials.gov # NCT03658642.


Comparing Lookback Periods to Ascertain Alzheimer's Disease and Related Dementias.
Claims data are a valuable resource for studying Alzheimer's disease and related dementias (ADRD). Alzheimer's disease and related dementias is often identified using a list of claims codes and a fixed lookback period of 3 years of data. However, a 1-year lookback or an approach using all-available lookback data could be beneficial based on different research questions. Thus, the purpose of this study was to compare 1-year and all-available lookback approaches to ascertaining ADRD compared to the standard 3-year approach. Using a cohort of Veterans hospitalized for heart failure (N = 373, 897), our results suggested high agreement (93% or greater) between the lookback periods. The 1-year lookback period had lower sensitivity (60%) and underestimated the prevalence of ADRD. These results suggest that 1-year and all-available lookback periods are viable approaches when using claims data.


Basal NO locally modulates human iliac artery function in vivo.
We demonstrated previously that endogenous NO influences large-artery distensibility in the ovine hindlimb. However, the role of basal NO in larger human conduit arteries is controversial. The aim of this study was to investigate whether basal production of NO, acting locally, influences iliac artery distensibility in humans. Distensibility was assessed by intra-arterial measurement of the pulse wave velocity. Eighteen subjects, free of significant coronary or iliac artery disease, were studied after diagnostic cardiac catheterization. Simultaneous pressure waveforms were recorded with a high-fidelity dual-pressure sensing catheter, placed in the common iliac artery during intra-arterial infusion of saline (baseline), glyceryl trinitrate (4 nmol/min), or NG-monomethyl-L-arginine (8 and 16 micromol/min). Drugs were infused proximally, via the catheter to perfuse the segment of artery under study, or distally, via the sheath, to control for any reflex changes in flow or sympathetic activation. Velocity was calculated using the foot-to-foot methodology. Six subjects received glyceryl trinitrate and 12 NG-monomethyl-L-arginine. There was no change in velocity after infusion of glyceryl trinitrate or NG-monomethyl-L-arginine via the sheath. However, infusion of glyceryl trinitrate via the catheter significantly reduced velocity by 31.43+/-5.80% (mean+/-SEM; P<0.01; P=0.02 for comparison). Likewise, infusion of the highest dose of NG-monomethyl-L-arginine via the catheter significantly increased velocity by 27.25+/-8.20% (P=0.001; P=0.02 for comparison). Importantly, there was no change in mean arterial blood pressure throughout the studies. These data indicate that under resting conditions, local NO production modulates human iliac artery distensibility and that exogenous NO increases arterial distensibility.


Biomechanical and microstructural characteristics of a collagen film-based corneal stroma equivalent.
The growth in refractive surgeries and corneal replacements has fueled interest in the development of a tissue-engineered cornea. This study characterizes the microstructure and biomechanical properties of film-based corneal stroma equivalents over time in culture. The increased collagen density in the films was hypothesized to result in improved mechanical properties both initially and over time. The microstructure of the film-based stromal equivalent was examined using atomic force microscopy and scanning electron microscopy; the mechanical properties, relaxed modulus, and ultimate tensile strength were quantified using uniaxial tensile testing. The dense, film-based stromal equivalent had a lamellae-like microstructure, which was notably different than the porous structure of sponges used previously. Seeded human corneal stromal fibroblasts remained on the surface of the film rather than migrating into the film and produced fibers of extracellular matrix with diameters of 35-75 nm. After an initial decrease during hydration, the relaxed modulus and ultimate tensile strength for fully hydrated collagen films were 0.4 +/- 0.2 MPa and 0.3 +/- 0.1 MPa, respectively. The mechanical properties of cell-seeded films mimicked those of control films. While further studies are needed to quantify the optical properties, the dense, lamellae-like structure of collagen films is a feasible scaffold for the development of tissue-engineered stroma.


Sex modulates hepatic mitochondrial adaptations to high-fat diet and physical activity.
The impact of sexual dimorphism and mitophagy on hepatic mitochondrial adaptations during the treatment of steatosis with physical activity are largely unknown. Here, we tested if deficiencies in liver-specific peroxisome proliferative activated-receptor-γ coactivator-1α (PGC-1α), a transcriptional coactivator of biogenesis, and BCL-2/ADENOVIRUS EIB 19-kDa interacting protein (BNIP3), a mitophagy regulator, would impact hepatic mitochondrial adaptations (respiratory capacity, H2O2 production, mitophagy) to a high-fat diet (HFD) and HFD plus physical activity via voluntary wheel running (VWR) in both sexes. Male and female wild-type (WT), liver-specific PGC-1α heterozygote (LPGC-1α), and BNIP3 null mice were thermoneutral housed (29-31°C) and divided into three groups: sedentary-low-fat diet (LFD), 16 wk of (HFD), or 16 wk of HFD with VWR for the final 8 wk (HFD + VWR) (n = 5-7/sex/group). HFD did not impair mitochondrial respiratory capacity or coupling in any group; however, HFD + VWR significantly increased maximal respiratory capacity only in WT and PGC-1α females. Males required VWR to elicit mitochondrial adaptations that were inherently present in sedentary females including greater mitochondrial coupling control and reduced H2O2 production. Females had overall reduced markers of mitophagy, steatosis, and liver damage. Steatosis and markers of liver injury were present in sedentary male mice on the HFD and were effectively reduced with VWR despite no resolution of steatosis. Overall, reductions in PGC-1α and loss of BNIP3 only modestly impacted mitochondrial adaptations to HFD and HFD + VWR with the biggest effect seen in BNIP3 females. In conclusion, hepatic mitochondrial adaptations to HFD and treatment of HFD-induced steatosis with VWR are more dependent on sex than PGC-1α or BNIP3.


Expanding the role of nurses in rehabilitation.
The role of nurses in geriatric rehabilitation is an aggregate of responsibilities directed toward planning realistic and meaningful restorative measures for disabled elderly persons. Nurse practitioners in this field are challenged to provide intelligent, competent bedside care, but it is equally important that they strive to create a milieu in which meaningful stimulation and sensory input are deliberately introduced. The geriatric rehabilitation nurse may be defined as a jack-of-all-trades and as a master of innovation. In the absence of a physical, occupational, or speech therapist, the nurse should be able to institute safe and stimulating activity programs. In the absence of a social worker, the nurse should have the interviewing and counseling skills that will enable intelligent intervention with helpful guidance in a social crisis.


Circadian rhythms, athletic performance, and jet lag.
Rapid air travel across several time zones exposes the traveller to a shift in his/her internal biological clock. The result is a transient desynchronisation of the circadian rhythm, called jet lag, lasting until the rhythm is rephased to the new environmental conditions. The most commonly experienced symptoms are sleep disorders, difficulties with concentrating, irritability, depression, fatigue, disorientation, loss of appetite, and gastrointestinal disturbance. Apart from the decrements in mental and physical performance directly consequent on such symptoms, competitive athletes are also exposed to the additional negative consequences of a shift from the optimal circadian window of performance. A brief summary of the possible negative effects of jet lag on athletic performance and potentially alleviating strategies is given.


Two T-box genes play independent and cooperative roles to regulate morphogenesis of ciliated Kupffer's vesicle in zebrafish.
The brain, heart and gastro-intestinal tract develop distinct left-right (LR) asymmetries. Asymmetric cilia-dependent fluid flow in the embryonic node in mouse, Kupffer's vesicle in zebrafish, notochordal plate in rabbit and gastrocoel roof plate in frog appears to be a conserved mechanism that directs LR asymmetric gene expression and establishes the orientation of organ asymmetry. However, the cellular processes and genetic pathways that control the formation of these essential ciliated structures are unknown. In zebrafish, migratory dorsal forerunner cells (DFCs) give rise to Kupffer's vesicle (KV), a ciliated epithelial sheet that forms a lumen and generates fluid flow. Using the epithelial marker atypical Protein Kinase C (aPKC) and other markers to analyze DFCs and KV cells, we describe a multi-step process by which DFCs form a functional KV. Using mutants and morpholinos, we show that two T-box transcription factors-No tail (Ntl)/Brachyury and Tbx16/Spadetail-cooperatively regulate an early step of DFC mesenchyme to epithelial transition (MET) and KV cell specification. Subsequently, each transcription factor independently controls a distinct step in KV formation: Tbx16 regulates apical clustering of KV cells and Ntl is necessary for KV lumen formation. By targeting morpholinos to DFCs, we show that these cell autonomous functions in KV morphogenesis are necessary for LR patterning throughout the embryo.


A rare sequence variant in intron 1 of THAP1 is associated with primary dystonia.
Although coding variants in THAP1 have been causally associated with primary dystonia, the contribution of noncoding variants remains uncertain. Herein, we examine a previously identified Intron 1 variant (c.71+9C>A, rs200209986). Among 1672 subjects with mainly adult-onset primary dystonia, 12 harbored the variant in contrast to 1/1574 controls (P < 0.01). Dystonia classification included cervical dystonia (N = 3), laryngeal dystonia (adductor subtype, N = 3), jaw-opening oromandibular dystonia (N = 1), blepharospasm (N = 2), and unclassified (N = 3). Age of dystonia onset ranged from 25 to 69 years (mean = 54 years). In comparison to controls with no identified THAP1 sequence variants, the c.71+9C>A variant was associated with an elevated ratio of Isoform 1 (NM_018105) to Isoform 2 (NM_199003) in leukocytes. In silico and minigene analyses indicated that c.71+9C>A alters THAP1 splicing. Lymphoblastoid cells harboring the c.71+9C>A variant showed extensive apoptosis with relatively fewer cells in the G2 phase of the cell cycle. Differentially expressed genes from lymphoblastoid cells revealed that the c.71+9C>A variant exerts effects on DNA synthesis, cell growth and proliferation, cell survival, and cytotoxicity. In aggregate, these data indicate that THAP1 c.71+9C>A is a risk factor for adult-onset primary dystonia.


Safety and immunogenicity of the malaria vaccine candidate GMZ2 in malaria-exposed, adult individuals from Lambaréné, Gabon.
Malaria is still one of the major public health threats in sub-Saharan Africa. An effective vaccine could be a sustainable control measure that can be integrated into existing health infrastructures. The malaria vaccine candidate GMZ2 is a recombinant fusion protein of conserved parts of Plasmodium falciparum Glutamate Rich Protein and Merozoite Surface Protein 3 adjuvanted with aluminium hydroxide. GMZ2 is immunogenic and well tolerated in malaria-naive adults from Germany. To assess safety and immunogenicity in malaria-exposed individuals, 40 adults from Lambaréné, Gabon were randomly assigned to receive either 100 μg GMZ2 or a rabies control vaccine three times in monthly intervals. Both vaccines were well tolerated. One month after a full course of vaccination, GMZ2-vaccinated individuals had 1.4-fold (95% confidence interval: [1.1, 1.7]) higher baseline-corrected anti-GMZ2 antibody levels and more GMZ2-specific memory B-cells compared to the rabies group (p=0.039), despite a high prevalence of GMZ2-specific immune reactivity due to previous intense exposure to P. falciparum.


Clinicopathological Characteristics of Superficial Barrett's Adenocarcinoma in a Japanese Population: A Retrospective, Multicenter Study.
Objective Although Barrett's adenocarcinoma (BA) remains a minor disease in Japan, its incidence has been gradually increasing. We analyzed the characteristics of BA in Japanese populations. Methods We retrospectively reviewed medical records and analyzed the clinicopathological differences between short-segment Barrett's esophagus (SSBE) and long-segment Barrett's esophagus (LSBE), as well as metastasis. Local recurrence and metachronous lesions were analyzed only in patients who underwent endoscopic resection (ER). Patients Consecutive patients who had pathological T1 BAs resected by ER or surgery from January 2003 to December 2017. Results A total of 168 patients were analyzed, including 139 with SSBE and 29 with LSBE. In total, 67% of the SSBE lesions and 32% of the LSBE lesions were located between 0 and 3 o'clock (p=0.0014). No patients who achieved pathological margin-free resection (pR0) and 17% of patients who did not achieve pR0 experienced local recurrence (p=0.0131). None of the patients without lymphovascular involvement, a poorly differentiated component, lesion size of >30 mm, and submucosal invasion of >500 μm experienced metastasis. The 5-year cumulative incidence rate of metachronous BA after ER was 0% in patients with SSBE and 40% in patients with LSBE (p=0.0005). Conclusion Superficial BA was likely to be detected at the right anterior wall of SSBE in the Japanese population. The risk for metachronous BA after ER was high in Japanese patients with LSBE, as in Western patients.


PAK4 and alphaPIX determine podosome size and number in macrophages through localized actin regulation.
Podosomes are actin-rich adhesion structures typical for monocytic cells and are implicated in migration and invasion. Major modes of podosome regulation include RhoGTPase signaling and actin regulatory pathways. However, it is not clearly understood how these signals induce highly localized changes in podosome formation and dynamics. Here, we show that the RhoGTPase effector PAK4, a member of the p21 associated kinase family, and its regulator alphaPIX (PAK-interacting exchange factor), are central to podosome formation in primary human macrophages. Immunofluorescence, biochemical and microarray data indicate that PAK4 acts as physiological regulator of podosomes in this system. Accordingly, transfection of a specific shRNA, as well as expression of PAK4 truncation mutants, resulted in reduced numbers of podosomes per cell. Moreover, expression of kinase active or inactive PAK4 mutants enhanced or reduced the size of individual podosomes, respectively, indicating a modulatory influence of PAK4 kinase activity on podosome size. Similar to the results gained with PAK4, cellular/overexpressed PIX was shown to be closely associated with podosomes. Moreover, both overexpression of alphaPIX wt and a mutant lacking the SH3 domain led to coalescence of podosomes. In sum, we propose that PAK4 and alphaPIX can induce highly localized changes in actin dynamics and thereby regulate size and number of podosomes in primary human macrophages.


[The epidemiology of malaria in Elazig between 1996 and 2004].
Malaria is a very important disease both for the world and Turkey. In this retrospective study, we investigated malaria using the data provided by The Health Administration of Elazig between 1996 and 2004. A total of 79,458 blood smears were examined during this 9-year period and malaria was found in 200 (0.25%). Except for one case of foreign origin infected with Plasmodium malaria, all of the cases were caused by Plasmodium vivax. All patients were imported cases who had come to Elazig from other regions for different reasons. We hope that our study contributes to the epidemiology of malaria and its prevention.


Excretion of the principal urinary metabolites of phenytoin and absolute oral bioavailability determined by use of a stable isotope in patients with epilepsy.
The anticonvulsant properties of phenytoin (PHT) were discovered in 1938. Since then, it has been one of the most widely used antiepileptic drugs. It is slowly absorbed, extensively plasma protein-bound, exhibits a nonlinear, concentration-dependent pharmacokinetic profile, and has a narrow therapeutic range. We determined PHT bioavailability during steady-state therapy by 1) measurement of the two principal deconjugated PHT urinary metabolites, 5-(4-hydroxyphenyl)-5-phenylhydantoin (p-HPPH) and 5-(3,4-dihydroxy-1,5-cyclohexadien-1-yl)-5-phenylhydantoin (DHD); and 2) direct determination of absolute bioavailability after simultaneous administration of an oral formulation and parenteral stable-labeled PHT (SL-PHT). Urinary metabolites were quantified by an isocratic HPLC-NI-APCI-MS method. The urinary dose recovery was calculated by dividing the molar recovery of the major PHT urinary metabolites by the molar dose received. Urinary metabolite recovery was surprisingly low, 35.4% ± 15.7% in younger patients (age 21-49 years old) and 32.9% ± 15.0% in patients aged 65 to 93 years. Absolute bioavailability was 86.4% ± 19.4% and 92.5% ± 25.2%, respectively. A weak, but significant, Spearman rank correlation was observed between urinary metabolite recovery and oral bioavailability (P = 0.00924, R = 0.166). This weak correlation may be the result of variability in urinary versus biliary-fecal excretion of p-HPPH glucuronide. This study demonstrates that daily PHT oral absorption exhibits wide interpatient variability, which may account for fluctuations in PHT concentration over time.


Delivering an enabling environment and multiple benefits for land degradation neutrality: Stakeholder perceptions and progress.
Achieving land degradation neutrality (LDN) was adopted by countries in 2015 as one of the targets of the global Sustainable Development Goals (SDGs). As LDN is a relatively new concept there is an increasing need for evidence on the potential socio-economic and environmental benefits of LDN as well as how an enabling environment for implementing LDN measures can be developed. This paper summarises the results from a global survey of LDN stakeholders, and a review of national progress in target setting that was commissioned by the United Nations Convention to Combat Desertification (UNCCD) in 2018. The study presents the perceptions of relevant stakeholders on the key components of an enabling environment for achieving and maintaining LDN (institutional, financial, policy/regulatory, and science-policy) as well as expectations of multiple benefits from its implementation. We also highlight key challenges and gaps in progress to date that are emerging from ongoing national target setting programs to implement LDN. The study finds that progress in implementing LDN has been widespread across countries. However there remains a lack of awareness of LDN and its key concepts along with high-level political buy-in. This may be impeding the integration of LDN into national development planning and budgeting processes where progress was assessed as limited. National capacities for securing land tenure and governance arrangements and integrated land use planning were perceived as comparatively low, further hampering the implementation of LDN. Despite these gaps, most stakeholders (>90 %) who participated in the global survey expected LDN to deliver a broad range of multiple benefits for human wellbeing, livelihoods and the natural environment. We argue that greater efforts are needed to raise awareness of LDN, educate core stakeholders in its concepts, enablers and benefits, raise its political profile, and provide evidence on national measures that will support implementation of LDN.


Effect of organizational and environmental factors on service differentiation strategy of integrated healthcare networks.
During the past decade, the missions/goals of medical providers of healthcare services in the United States have shifted--from emphasizing individual, independent illness treatments to focusing on the continuum of care, population-based wellness, and providing the appropriate care in the most efficient way. Integrated healthcare networks (IHNs)--or integrated healthcare delivery systems--have been focusing heavily on their level of various partnership integration (i.e. service differentiation strategy) in order to offer a full continuum of care. The aim of this study, using the individual IHN as the unit of analysis, was to identify organizational and environmental factors that influence IHN administrators to focus on their service differentiation of market lines, including the establishment of third-party payers' contracts, the affiliation of managed-care organizations, and the alliances of various nonhospital medical providers, to provide a continuum of care. The study findings show that tax status of an IHN, its age, and market competition affect its service differentiation strategy in the provision of a full continuum of care.


[Polyneuropathy after cisplatin treatment].
Polyneuropathy developed in 7 of 87 women who had been treated with cis-platinum for ovarian carcinoma. Distal disturbances of sensitivity were clinically prominent, the emphasis being on disturbed vibratory perception and acrognosis. No relevant signs of motor dysfunction were noticed, neither clinically nor neurophysiologically. Sural biopsies taken from two women revealed a primary axonal degeneration, mostly affecting the myelinic fibres of large diameter. In 3 cases mild reversion was seen, and in 4 cases no definite reversion, of the disturbed functions - uncertain gait and clumsiness of the hands - during the follow-up period of 6 to 30 months.


DTIC (NSC-45388) studies in the southwest oncology group.
In a phase I study, the best antitumor/toxicity ratio for DTIC was reported to be at a dose of 250 mg/m2/day X 5 repeated at 28-day intervals. Nausea, vomiting, leukopenia, and thrombocytopenia were the major toxic effects noted. The best responses were seen in disseminated melanoma (19%), various sarcomas (22%), and Hodgkin's disease. A subsequent phase II study in refractory lymphomas showed a response rate in Hodgkin's disease of 56%. In disseminated melanomas, DTIC was then combined with vincristine and BCNU and demonstrated a response rate of 23% which did not improve with the addition of chlorpromazine (23%). A response rate of 31% was seen with the combination of DTIC, BCNU, and hydroxyurea which did not improve with the addition of vincristine (30%). Responders had a more significant survival rate as compared to nonresponders.


Hypophosphatemia is an independent risk factor for AKI among hospitalized patients with COVID-19 infection.
This study sought to investigate incidence and risk factors for acute kidney injury (AKI) in hospitalized COVID-19. In this retrospective study, we enrolled 823 COVID-19 patients with at least two evaluations of renal function during hospitalization from four hospitals in Wuhan, China between February 2020 and April 2020. Clinical and laboratory parameters at the time of admission and follow-up data were recorded. Systemic renal tubular dysfunction was evaluated via 24-h urine collections in a subgroup of 55 patients. In total, 823 patients were enrolled (50.5% male) with a mean age of 60.9 ± 14.9 years. AKI occurred in 38 (40.9%) ICU cases but only 6 (0.8%) non-ICU cases. Using forward stepwise Cox regression analysis, we found eight independent risk factors for AKI including decreased platelet level, lower albumin level, lower phosphorus level, higher level of lactate dehydrogenase (LDH), procalcitonin, C-reactive protein (CRP), urea, and prothrombin time (PT) on admission. For every 0.1 mmol/L decreases in serum phosphorus level, patients had a 1.34-fold (95% CI 1.14-1.58) increased risk of AKI. Patients with hypophosphatemia were likely to be older and with lower lymphocyte count, lower serum albumin level, lower uric acid, higher LDH, and higher CRP. Furthermore, serum phosphorus level was positively correlated with phosphate tubular maximum per volume of filtrate (TmP/GFR) (Pearson r = 0.66, p < .001) in subgroup analysis, indicating renal phosphate loss via proximal renal tubular dysfunction. The AKI incidence was very low in non-ICU patients as compared to ICU patients. Hypophosphatemia is an independent risk factor for AKI in patients hospitalized for COVID-19 infection.


Skin Inflammation and Testicular Function: Dermatitis Causes Male Infertility via Skin-Derived Cytokines.
The medical comorbidities including skin diseases are associated with male infertility. The most common cause of male infertility is the inability of testes to produce sperm; however, the influence of persistent dermatitis on testicular function has not been elucidated so far. We investigated the relationship between skin inflammation and impaired sperm production using a spontaneous dermatitis mouse model. We examined the breeding records of dermatitis mice and their wild-type littermates. Sperm count, motility, and viability were analyzed by direct microscopic observation and flow cytometry. In addition, testis and epididymis were histologically examined. Finally, sperm viability was evaluated in another dermatitis mouse model and in wild-type mice in which inflammatory cytokines were intraperitoneally administered. Compared to wild-type littermate mice, the number of children born was lower in mice with dermatitis. The body weight and testis size were decreased age-dependently. In the skin disease group, the sperm count and movement ratio were clearly decreased, and reduced sperm viability was observed. Histological examination revealed the detachment of Sertoli cells and reduced spermatogenesis. The fibrosis of epididymal stroma was severe, and it might affect defective sperm maturation in the epididymis. In addition, this phenomena was reproduced by a hapten applied dermatitis mouse model and the intraperitoneal administration of inflammatory cytokines. Once the skin is inflamed, inflammatory cytokines are produced and released, which may affect testicular and sperm function. Additional studies are needed to determine the relationship between male infertility and severe dermatitis in human.


The Use of Inflammatory Markers for Treatment Response Monitoring in Acute Cholangitis: A Retrospective Cohort Study.
In acute cholangitis (AC), monitoring treatment response to antimicrobial therapy allows for making timely decisions on early biliary decompression. The aims of this study were to compare the discriminating powers of traditional blood inflammatory markers and propose new inflammatory markers that have a better ability to distinguish between patients with and without biliary tract infection. This was a retrospective cohort study. Patients who underwent endoscopic retrograde cholangio-pancreaticography for AC and those without biliary tract inflammation were randomly selected in the 4:3 ratio of their hospital admissions from our hospital endoscopic retrograde cholangio-pancreaticography database. The exclusion criterion was the absence of C-reactive protein (CRP) measurements. The discriminating powers of the neutrophil count, lymphocytes, albumin, neutrophil-to-lymphocyte ratio, and CRP were superior to that of white blood cell (P1 < 0.005; P2 = 0.004; P3 < 0.0005; P4 < 0.0005; P5 < 0.0005). In monitoring treatment response in AC, lymphocyte count, albumin, neutrophil-to-lymphocyte ratio, and CRP were better than neutrophil count (P6 = 0.037, P7 < 0.005, P8, 9 < 0.0005). The area under the receiver operating characteristic curve (AUC) of CRP was higher than the AUC for lymphocytes, 96% (95% confidence interval [CI]: 94-98%) versus 81% (95% CI: 76-86%) (P < 0.0005), and larger than the AUC for albumin, 88% (95% CI: 84-92%) (P < 0.0005), indicating a greater discriminating power of CRP. However, the discriminating power of CRP-to-lymphocyte ratio (CLR) was more than that for CRP (P = 0.006) but equal to CRP-to-(lymphocytes∗albumin) ratio (CLAR) (P = 0.249). The AUCs of CLR and CLAR were both 98% (95% CI: 96-99%). CLR and CLAR have superior discriminating powers than traditional inflammatory markers used for monitoring treatment response in AC.


An indirect fluorescent antibody technique for Haemophilus ducreyi.
An indirect fluorescent antibody technique which allows the detection of antibody against or the recognition of Haemophilus ducreyi has been developed. Heat inactivated antiserum was reacted uith prepared smears of Haimophilus species and selected organisms commonly associated with penile ulcerations. In serum absorbed with Haemophilus sp no fluorescence was observed with 156 heterologous organisms, but all six strains of bacteriologically confirmed H. ducreyi gave significant reactions with the absorbed anti-WD-68 serum.


Brief report: the burden of diabetes therapy: implications for the design of effective patient-centered treatment regimens.
Management of diabetes, and in particular blood glucose, can be complex and burdensome. To evaluate patient views of the burdens of therapy and its impact on self-management. Veteran patients with type 2 diabetes. Mailed survey. Patients described their views of the burden of diabetes treatments, adherence, and clinical and demographic status. Factors associated with ratings of burden and adherence to therapy were examined using multivariate regression methods. The response rate was 67% (n=1,653). Patients viewed pills as the least burdensome treatment and insulin as the most burdensome. Ratings of the burden of insulin were lower if a patient had prior experience with therapy. Adherence to prescribed therapy varied substantially; for example, patients followed medication recommendations more closely than other areas of self-management. Multivariate analyses showed that the main predictor of adherence was patients' ratings of the burden of therapy. Injected insulin regimens are viewed as highly burdensome by patients, although this burden is attenuated by experience. Adherence to self-management is strongly and independently correlated with views of treatment burden. The burden of diabetes-related treatments may be a source of suboptimal glucose control seen in many care settings. Providers should consider the burden of treatment for a particular patient and its impact on adherence as part of a decision-making process to design effective treatment regimens.


Neu antigen-negative variants can be generated after neu-specific antibody therapy in neu transgenic mice.
Prolonged administration of HER-2/neu-specific monoclonal antibody therapy is now widely used for the treatment of HER-2/neu-overexpressing tumors in advanced-stage breast cancer patients. Monoclonal antibody therapy has the potential to promote reduced tumor expression of HER-2/neu by receptor down-modulation and/or the generation of antigen-negative variants. Loss of antigen by either mechanism could potentially impact subsequent therapeutic strategies targeting HER-2/neu. In this study, the effects of chronic neu-specific monoclonal antibody therapy on tumor growth and neu protein expression were examined in a murine model of neu-overexpressing breast cancer. Treatment of neu-overexpressing tumors with neu-specific antibody, in vitro or in vivo, resulted in significant tumor growth inhibition. When neu antibody was used to treat neu-overexpressing tumor cells both in vitro and in vivo in tumor-bearing mice, neu receptor expression was not diminished after cessation of therapy. However, in the setting of clinically undetectable disease in a fraction of animals, antigen-negative variants were generated. An understanding of the effects of monoclonal antibodies on target antigen expression is critical for the future design and testing of novel HER-2/neu-targeted therapies administered in combination with or after HER-2/neu-specific monoclonal antibody therapy.


In Vitro Antifungal Susceptibility of Yeast and Mold Phases of Isolates of Dimorphic Fungal Pathogen Emergomyces africanus (Formerly Emmonsia sp.) from HIV-Infected South African Patients.
Disseminated emmonsiosis is an important AIDS-related mycosis in South Africa that is caused by Emergomycesafricanus, a newly described and renamed dimorphic fungal pathogen. In vitro antifungal susceptibility data can guide management. Identification of invasive clinical isolates was confirmed phenotypically and by sequencing of the internal transcribed spacer region. Yeast and mold phase MICs of fluconazole, voriconazole, itraconazole, posaconazole, caspofungin, anidulafungin, micafungin, and flucytosine were determined with custom-made frozen broth microdilution (BMD) panels in accordance with Clinical and Laboratory Standards Institute recommendations. MICs of amphotericin B, itraconazole, posaconazole, and voriconazole were determined by Etest. Fifty unique E. africanus isolates were tested. The yeast and mold phase geometric mean (GM) BMD and Etest MICs of itraconazole were 0.01 mg/liter. The voriconazole and posaconazole GM BMD MICs were 0.01 mg/liter for both phases, while the GM Etest MICs were 0.001 and 0.002 mg/liter, respectively. The fluconazole GM BMD MICs were 0.18 mg/liter for both phases. The GM Etest MICs of amphotericin B, for the yeast and mold phases were 0.03 and 0.01 mg/liter. The echinocandins and flucytosine had very limited in vitro activity. Treatment and outcome data were available for 37 patients; in a multivariable model including MIC data, only isolation from blood (odds ratio [OR], 8.6; 95% confidence interval [CI], 1.3 to 54.4; P = 0.02) or bone marrow (OR, 12.1; 95% CI, 1.2 to 120.2; P = 0.03) (versus skin biopsy) was associated with death. In vitro susceptibility data support the management of disseminated emmonsiosis with amphotericin B, followed by itraconazole, voriconazole, or posaconazole. Fluconazole was a relatively less potent agent.


S100B concentrations increase perioperatively in jugular vein blood despite limited metabolic and inflammatory response to clinically uneventful carotid endarterectomy.
Our aim was to test the hypothesis that metabolic and inflammatory responses of the brain perioperatively during carotid endarterectomy (CEA) might affect blood brain barrier (BBB) integrity. Twenty patients with >70% stenosis of internal carotid artery (ICA) were prospectively included. Surgery was performed under general anaesthesia. Blood was sampled from ipsilateral internal jugular vein and radial artery: just before, during, and after ICA clamping S100B protein, glucose, lactate, 20 amino acids, and key cytokines were analysed. Jugular vein S100B increased during clamping and reperfusion, while a marginal systemic increase was recorded, unrelated to stump pressure during clamping. Glucose increased during clamping in jugular vein blood and even more systemically, while jugular lactate values were higher than systemic values initially. Most amino acids did not differ significantly between jugular vein and systemic levels: glutamic acid and aspartic acid decreased during surgery while asparagine increased. Jugular vein interleukin (IL)-6 showed a transient non-significant increase during clamping and decreased systemically. IL-8 and IL-10 increased over time. Rising jugular vein S100B concentrations indicated reduced BBB integrity, and marginal secondary increase of S100B systemically. Limited ischaemic effects on the brain during cross-clamping, unrelated to S100B concentrations, were confirmed by lower brain glucose levels and higher lactate levels than in systemic blood. The lack of increased jugular vein glutamic acid disproves any major ischaemic brain injury following CEA. The inflammatory response was limited, did not differ greatly between jugular and systemic blood, and was unrelated to S100B.


Left Atrial Strain Predicts Stroke Recurrence and Death in Patients With Cryptogenic Stroke.
Left atrial strain (LAS) has been widely studied as a predictor of atrial fibrillation (AF) after cryptogenic stroke (CS). However, the evidence about its prognostic role in terms of stroke recurrence and death in this setting remains scarce. A total of 92 consecutive patients with ischemic stroke or transient ischemic attack with ABCD2 scale ≥4 of unknown etiology were prospectively recruited. Echocardiography, including LAS was performed during admission. The primary outcome measure was the composite of stroke recurrence or death. The mean age was 77.5 ± 7.7, and 58% of patients were female. After a median follow up of 28 months, the primary outcome measure occurred in 15 patients (16%). The primary outcome was more frequent in patients with diabetes (53% vs 21%, p = 0.02), chronic kidney disease (33% vs 10%, p = 0.034), and a history of heart failure (13% vs 0%, p = 0.025). LAS reservoir (LASr) and LAS conduit (LAScd) were lower in patients developing the primary outcome (21% ± 7% vs 28.8% ± 11%, p = 0.017 and 7.7% ± 3.9% vs 13.7% ± 7%, p = 0.007, respectively). On multivariate analysis, LASr (hazard ratio 0.9, 95% confidence interval 0.85 to 0.99, p = 0.048) and diabetes (hazard ratio 3.3, 95% confidence interval 1.03 to 10.4, p = 0.045) were associated with stroke recurrence or all-cause death after CS. On the log-rank test (using the discriminatory cut-off value of LASr <23%), LASr (p = 0.009) was associated with higher risk of the primary outcome. In conclusion, lower values of the LAS reservoir were associated with a higher risk of stroke recurrence or death after CS. LAS may identify patients at higher risk of thromboembolism and stress conditions.


[Measurement of proinsulin, insulin and C-peptide in human blood by high performance liquid chromatography].
The technique of measuring insulin, proinsulin, and C-peptide in human blood by high-pressure liquid chromatography is described. Donors, subjects with abnormal glucose tolerance, and patients with type I diabetes were examined. The main advantage of the method over radioimmunoassay and enzyme immunoassay is the possibility of simultaneous differentiated measurement of human insulin and animal hormone administered parenterally, and separation of insulin and its precursor on the basis of difference in molecular weight and charge of molecules. The results indicate that patients with type I diabetes retain the secretion of endogenous insulin, although its level is appreciably lower than in healthy donors, and the structure of its molecules is heterogeneous, which is proven by a biphasic chromatographic peak. Inhibition of proinsulin transformation into insulin is one probable mechanism of impairment of glucose tolerance. High-pressure liquid chromatography can be used at clinical laboratories for examinations of diabetics.


The work of ship's doctors of Polish Ocean Lines.
In 1984, during 201 voyages of cargo ships of Polish Ocean Lines having doctors on board, 30340 medical consultations were given to crew members, in 15506 cases of complaints and diseases. Doctors also consulted injured seafarers 2456 times in cases of injuries, wounds, contusions, broken bones, burns, scalds etc. During these 201 voyages, 33 cases of serious medical conditions were reported among crew members, which required disembarkation and hospital treatment abroad; in this number two cases of myocardial infarction ended fatally. There were also 13 casualties among seamen, which were treated in hospitals in foreign ports; all of them survived. One of the important health problems of seafarers were diseases of teeth, pulp, and periapical tissue. Such cases constituted the majority (66.9%) of all 1868 cases referred for consultation and treatment in foreign ports. Oral diseases prevention among seafarers needed improvement. The working time of doctors on board ship was under-utilized. To a greater extent they should be involved in training seafarers in first aid and primary health care, and in health education activities among seamen, during sea voyages. Materials presented in the paper may be of use in planning efficient and cost-effective health services for crews of ships.


Quantitative Proteomic Analysis of Optimal Cutting Temperature (OCT) Embedded Core-Needle Biopsy of Lung Cancer.
With recent advances in understanding the genomic underpinnings and oncogenic drivers of pathogenesis in different subtypes, it is increasingly clear that proper pretreatment diagnostics are essential for the choice of appropriate treatment options for non-small cell lung cancer (NSCLC). Tumor tissue preservation in optimal cutting temperature (OCT) compound is commonly used in the surgical suite. However, proteins recovered from OCT-embedded specimens pose a challenge for LC-MS/MS experiments, due to the large amounts of polymers present in OCT. Here we present a simple workflow for whole proteome analysis of OCT-embedded NSCLC tissue samples, which involves a simple trichloroacetic acid precipitation step. Comparisons of protein recovery between frozen versus OCT-embedded tissue showed excellent consistency with more than 9200 proteins identified. Using an isobaric labeling strategy, we quantified more than 5400 proteins in tumor versus normal OCT-embedded core needle biopsy samples. Gene ontology analysis indicated that a number of proliferative as well as squamous cell carcinoma (SqCC) marker proteins were overexpressed in the tumor, consistent with the patient's pathology based diagnosis of "poorly differentiated SqCC". Among the most downregulated proteins in the tumor sample, we noted a number of proteins with potential immunomodulatory functions. Finally, interrogation of the aberrantly expressed proteins using a candidate approach and cross-referencing with publicly available databases led to the identification of potential druggable targets in DNA replication and DNA damage repair pathways. We conclude that our approach allows LC-MS/MS proteomic analyses on OCT-embedded lung cancer specimens, opening the way to bring powerful proteomics into the clinic. Graphical Abstract .


Movements and residency of Caribbean reef sharks at a remote atoll in Belize, Central America.
We investigated spatial use patterns of 77 Caribbean reef sharks (Carcharhinus perezi) at Lighthouse Reef Atoll, Belize over 7 years using residency patterns, kernel density (KD) estimation and network analysis. We found a high degree individual variation in spatial use of the atoll, but there were significant differences in residency and activity space between sexes, with females being overall more resident. Ontogenetic shifts in movement and residency were largely limited to females, as the residency index increased and activity space estimates decreased as females matured, while for males there was no relationship between space use or residency and size. KD analysis revealed many mature females were highly resident to discrete locations, and average activity space of the intermediate-sized sharks was significantly larger than that of the adults, but not the smallest sharks. Markov chain analyses indicated that the southwestern portion of the atoll was the most important movement corridor for all sharks. Both the Blue Hole and Half Moon Caye Natural Monuments provide some protection for larger Caribbean reef sharks; however, a gear ban on longlines on the southwestern forereef between Long Caye and the channel entrance to the Blue Hole would maximize the benefits for all sharks.


Symbiosis of Aeromonas veronii biovar sobria and Hirudo medicinalis, the medicinal leech: a novel model for digestive tract associations.
Hirudo medicinalis, the medicinal leech, is applied postoperatively in modern medicine. Infections by Aeromonas occur in up to 20% of patients unless a preemptive antibiotic treatment is administered. The associated infections demonstrate the need for a better understanding of the digestive tract flora of H. medicinalis. Early studies reported the presence of a single bacterial species in the digestive tract and suggested that these bacteria were endosymbionts contributing to the digestion of blood. In this study, we cultivated bacteria from the digestive tract and characterized them biochemically. The biochemical test results identified the isolates as Aeromonas veronii biovar sobria. This species identification was supported by sequence comparison of a variable region of the genes coding for 16S rRNA. In a colonization assay, a rifampin-resistant derivative of a symbiotic isolate was fed in a blood meal to H. medicinalis. The strain colonized the digestive tract rapidly and reached a concentration similar to that of the native bacterial flora. For the first 12 h, the in vivo doubling time was 1.2 h at 23 degreesC. After 12 h, at a density of 5 x 10(7) CFU/ml, the increase in viable counts ceased, suggesting a dramatic reduction in the bacterial growth rate. Two human fecal isolates, identified as Aeromonas hydrophila and A. veronii biovar sobria, were also able to colonize the digestive tract. These data demonstrate that the main culturable bacterium in the crop of H. medicinalis is A. veronii biovar sobria and that the medicinal leech can be used as a model for digestive tract association of Aeromonas species.


Effects of growth hormone on insulin resistance and atherosclerotic risk factors in obese type 2 diabetic patients with poor glycaemic control.
We aimed to evaluate the combined effects of GH treatment and diet restriction on lipolysis and anabolism, insulin resistance and atherosclerotic risk factors in obese patients with type 2 diabetes mellitus (T2DM). This randomized, double-blind, placebo-controlled study included 24 obese T2DM patients (male : female = 12 : 12, mean age 53.7 +/- 7.2 years) with poor glycaemic control (fasting plasma glucose 10.673 +/- 1.121 mmol/l, HbA(1C) 9.9 +/- 2.3%). Sixteen of these patients were treated with recombinant human GH (1-1.5 units/day, 5 days/week) while undergoing diet restriction and exercise for 12 weeks. Anthropometric and bioelectrical impedance measurements were undertaken to determine the lean body mass and total body fat. Computed tomography (CT) was performed to estimate visceral and subcutaneous fat distribution at the umbilicus level and the muscle area of the midthigh. Insulin resistance was measured by the insulin tolerance test (ITT) and by the homeostasis model assessment of insulin resistance (HOMA-IR). The ratios VSR (visceral fat area/subcutaneous fat area) and VMR (visceral fat area/thigh muscle area) were significantly decreased in the GH-treated group compared to the control group. An increase in lean body mass was observed in the GH-treated group. Levels of total cholesterol, triglyceride, free fatty acid (FFA), fibrinogen, and plasminogen activator inhibitor-1 (PAI-1) were significantly decreased after GH treatment. Fasting glucose levels decreased similarly (P < 0.05 anova) in both groups during the treatment period. Fasting C-peptide levels significantly increased, whereas insulin levels significantly decreased, in the GH-treated group, but no changes were observed in the control group. The insulin sensitivity index (ISI) was significantly increased in the GH-treated group (1.3 +/- 1.4 vs. 1.9 +/- 1.0%/min, P < 0.05). GH treatment in obese T2DM patients with poor glycaemic control is beneficial in decreasing the amount of visceral fats, and may therefore result in improvements in insulin resistance, atherosclerotic risk factors and dyslipidaemia.


Classification of Targets and Distractors in an Audiovisual Attention Task Based on Electroencephalography.
Within the broader context of improving interactions between artificial intelligence and humans, the question has arisen regarding whether auditory and rhythmic support could increase attention for visual stimuli that do not stand out clearly from an information stream. To this end, we designed an experiment inspired by pip-and-pop but more appropriate for eliciting attention and P3a-event-related potentials (ERPs). In this study, the aim was to distinguish between targets and distractors based on the subject's electroencephalography (EEG) data. We achieved this objective by employing different machine learning (ML) methods for both individual-subject (IS) and cross-subject (CS) models. Finally, we investigated which EEG channels and time points were used by the model to make its predictions using saliency maps. We were able to successfully perform the aforementioned classification task for both the IS and CS scenarios, reaching classification accuracies up to 76%. In accordance with the literature, the model primarily used the parietal-occipital electrodes between 200 ms and 300 ms after the stimulus to make its prediction. The findings from this research contribute to the development of more effective P300-based brain-computer interfaces. Furthermore, they validate the EEG data collected in our experiment.


Stenting of culprit lesions in unstable angina leads to a marked reduction in plaque burden: a major role of plaque embolization? A serial intravascular ultrasound study.
Intravascular ultrasound (IVUS) studies have shown that a mechanism of plaque compression/embolization contributes toward the poststenting increase in lumen area. The aim of this IVUS study was to compare the mechanisms of lumen enlargement after coronary stenting in 54 consecutive patients with unstable angina (UA) (group 1) and 56 with stable angina (group 2) to verify whether plaque embolization plays a major role in the former. Both groups underwent the IVUS assessment (speed, 0.5 mm/sec) before the intervention and after stent implantation. The lumen area, the external elastic membrane area, and the plaque+media area (PA) were measured at 0.5-mm intervals. PA reduction in the lesion site was significantly greater in group 1 (-2.50+/-1.97 versus -0.53+/-1.43 mm2, P<0.001). After stenting, 47% of the lumen area increase in group 1 was obtained by means of PA reduction, and 53% was attributable to external elastic membrane area increase; the corresponding figures in group 2 were 13% and 87% (P<0.05). Decrease in PA after stenting was the only significant predictor of the MB fraction of creatinine kinase (CK-MB) release in a multiple regression model (P=0.047). Serial volumetric IVUS assessment revealed in UA lesions a marked poststenting reduction in plaque volume, which is significantly greater than in stable angina and is associated with postprocedural CK-MB release. The decrease in PA during the procedure predicts CK-MB release in a multiple regression model. These findings suggest that stent deployment is often associated with plaque embolization in patients with UA.


Rectosigmoid prolapse--A case report.
Many factors have been indentified as a possible cause of rectal prolaps. Despite the fact that it is not a life- threating condition, its clinical presentation varies, and sometimes it can present as an emergency. We presented a patient with prolapse of an unusually large segment of the rectosigmoid colon caused by chronic constipation, as an incarcerated segment repaired surgically. A 62-year-old female patient was referred to the Emergency Department in bad condition with severe pain in the perianal region. On examination a complete rectal prolaps as well as a part of sigmoid colon were found. Macroscopically, the prolapsed segment appeared edematous, livid, with ulcerations. An attempt to manually reduce prolapse failed, therefore resection of 50 cm of sigmoid colon with rectopexy had to be performed. No complications occurred and the patient was without symptoms six months later. Colonoscopy did not reveal any abnormality. Although the preoperative management and preparation of the patient was limited, emergancy surgical intervention for such a case was the strategy of choice due to magnitude of the prolapsing segment. It provided a successful and permenant solution.