Glial cells decipher synaptic competition at the mammalian neuromuscular junction.
It is now accepted that glial cells actively interact with neurons and modulate their activity in many regions of the nervous system. Importantly, modulation of synaptic activity by glial cells depends on the proper detection and decoding of synaptic activity. However, it remains unknown whether glial cells are capable of decoding synaptic activity and properties during early postdevelopmental stages, in particular when different presynaptic nerve terminals compete for the control of the same synaptic site. This may be particularly relevant because a major determinant of the outcome of synaptic competition process is the relative synaptic strength of competing terminals whereby stronger terminals are more likely to occupy postsynaptic territory and become stabilized while weaker terminals are often eliminated. Hence, because of their ability to decode synaptic activity, glial cells should be able to integrate neuronal information of competing terminals. Using simultaneous glial Ca(2+) imaging and synaptic recordings of dually innervated mouse neuromuscular junctions, we report that single glial cells decipher the strength of competing nerve terminals. Activity of single glial cells, revealed by Ca(2+) responses, reflects the synaptic strength of each competing nerve terminal and the state of synaptic competition. This deciphering is mediated by functionally segregated purinergic receptors and intrinsic properties of glial cells. Our results indicate that glial cells decode ongoing synaptic competition and, hence, are poised to influence its outcome.


[The multidisciplinary management of acute pancreatitis: a review of 244 cases].
Acute pancreatitis (AP) has still a high mortality rate; therefore the accuracy of the predictors of severity actually employed and the therapeutic choices are under debate. The aim of this work is to valuate the accuracy of the prognostic factors actually employed and the results of a multidisciplinary treatment of the AP. A consecutive series of 244 patients affected by AP (168 mild, 76 severe) is analysed. Mean age 64.4 years (range 17-94 years old). As regard as aetiology is concerned, 166 are biliary pancreatitis, 42 alcoholic, 27 idiopathic, 9 iatrogenic. Ranson's score, Balthazar criteria, and ASA are employed to assess the high-risk patients. The treatment is medical and in severe AP the patient is admitted in ICU. 80 patients affected biliary AP undergo an ERCP. An endoscopic papillotomy is performed in 55 cases and in 49 biliary stones are removed. 90 patients underwent a delayed colcistectomy. 11 patients underwent a surgical treatment: 9 for infected necrotic pancreatitis, 1 for a hemorrhagic shock, 1 for peritonitis. The overall mortality, the mortality related to the severity of the AP, the prognostic evaluation of the etiology, the ASA index, the Ranson and Balthazar scores are evaluated. The overall mortality rate is 2.8% (0.6% in the mild AP and 7.8% in the severe AP). In the surgical group the mortality rate is 18.1%. The Ranson's score is the only independent factor in the multivariate analysis (p<0.0374) while also the Balthazar classification is significant in the univariate analysis. As regard as the prognostic factors in AP are concerned, the Ranson's score and Balthazar criteria are reliable even if many other predictors of severity are tested to make progress in the early detection of the complications. As regard as the therapy is concerned, the endoscopic papillotomy and the endoscopic removal of the biliary stones is the gold standard in the treatment of the biliary AP. In the other cases of AP medical treatment shall be started as early as possible with intensive care management in the severe AP. In patients with infected pancreatic necrosis or haemorrhage or peritonitis, surgery is the only therapeutic choice while the surgical treatment in case of sterile pancreatic necrosis with MODS unresponsive to medical therapy is still under debate.


Chlorinated Water Modulates the Development of Colorectal Tumors with Chromosomal Instability and Gut Microbiota in Apc-Deficient Mice.
The gastrointestinal tract is continuously exposed to a variety of chemicals and commensal bacteria. Recent studies have shown that changes in gut microbial populations caused by chlorine or other chemicals in the drinking water influence the development of human colorectal cancer, although the mechanism of tumorigenesis in the gut epithelium is obfuscated by the diversity of microflora and complexity of the tumor microenvironment. In this regard, mouse models that recapitulate human colorectal cancer are an invaluable tool. In this study, we used two conditional adenomatous polyposis coli (Apc) knockout mouse models to investigate the effect of chlorinated water on tumorigenesis in the digestive tract. Mice with colon-specific carcinoma--caused by either chromosomal (CDX2P 9.5-NLS Cre;Apc(+/flox), abbreviated to CPC;Apc) or microsatellite (CDX2P9.5-G19Cre;Apc(flox/flox) and CDX2P9.5-G22Cre;Apc(flox/flox)) instability, respectively--were administered chlorinated (10.0 mg/L chlorine) or tap (0.7 mg/L chlorine) water and evaluated for colon polyp formation. In CPC;Apc mice given chlorinated drinking water, tumors tended to develop in the colon, whereas in those that drank tap water, tumors were mostly observed in the small intestine. There was no difference in the rate of tumor formation of CDX2P9.5-G19Cre;Apc(flox/flox) and CDX2P9.5-G22Cre;Apc(flox/flox) mice consuming chlorinated as compared to tap water, suggesting that microsatellite instability in the Apc gene does not significantly affect tumorigenesis. Chlorinated water altered the enteric environment by reducing the fecal populations of the obligatory anaerobes Clostridium perfringens and C. difficile, as well as species belonging to the Atopobium cluster, including Enterobacteriaceae and Staphylococcus sp., which was associated with colon tumorigenesis in CPC;Apc mice. These results suggest that differences in tumorigenesis among CPC;Apc mice consuming chlorinated versus tap water may be due to differences in gastrointestinal commensal populations.


Generation of mice with a conditional Lbh null allele.
Limb bud and heart (LBH) is a developmentally expressed, tissue-specific transcription cofactor in vertebrates that acts in the WNT signaling pathway, a genetic program critical for embryogenesis and adult tissue homeostasis. Aberrant gain-of-function of LBH is implicated in both human congenital disease and cancer. The normal physiological function of LBH has remained elusive owing to a lack of genetic loss-of-function models. Here, we have generated mice with a conditional null allele of Lbh by flanking exon 2 with loxP sites (Lbh(flox)). Homozygous Lbh(flox) and Lbh(loxP) mice, in which the Neo cassette was removed through FLPe-mediated recombination, were viable and fertile, indicating that these conditional Lbh alleles are fully functional. Lbh(loxP) mice were then crossed with a Rosa26-Cre line, resulting in ubiquitous deletion of exon 2 and abolishment of LBH protein expression. Mice homozygous for the Lbh null allele (Lbh(Δ)(2)) displayed normal embryonic development and postnatal growth with morphologies indistinguishable from wild-type littermates. However, mammary gland development, which occurs primarily after birth, was perturbed. Thus, the conditional Lbh allele will be a valuable tool to uncover the currently unknown tissue-specific roles of LBH in postnatal development and disease.


Evaluation of genetic factors involved in nocturnal electromyographic activity of masticatory muscles in twins.
The objectives of this study were to assess sleep bruxism events by directly recording electromyographic activity during sleep and to reveal the relative importance of genetic and environmental factors involved in sleep bruxism in twins. The subjects consisted of 108 twins (mean age 22.2 ± 6.4 years). Electromyographic activity of temporalis muscles during sleep was evaluated using a portable automatic sleep bruxism analyzer (Grindcare 3.0, Medotech A/S), and recordings were carried out for at least three consecutive nights. Quantitative genetic statistics based on structural equation modeling was utilized to estimate variance components. Monozygotic twin-pair correlation for the number of nocturnal electromyographic activities recorded in this study (r = 0.463, P = 0.009) was higher than dizygotic twin-pair correlation (r = 0.103, P = 0.725). The proportion of total phenotypic variance in the liability of sleep bruxism to attribute to genetic influences, related to the electromyographic activities, was 48 % (95 % CI 17-95 %) and to unique environmental influences was 52 % (95 % CI 28-82 %). Additive genetic effects may be a contributing factor to the occurrence of nocturnal EMG activity associated with sleep bruxism. A greater understanding of the contribution of genetic factors could have beneficial uses, including enhanced accuracy of sleep bruxism diagnosis, management of sleep bruxism, and enhanced estimation of the prognosis for patients suffering from sleep bruxism. In addition, it could be also important to adequately evaluate the environmental factors in patients with sleep bruxism.


Is ambulatory blood pressure monitoring cost-effective in the routine surveillance of treated hypertensive patients in primary care?
Achieving target levels of office and/or ambulatory blood pressure readings among treated hypertensive patients is an important aspect of cardiovascular disease prevention. Although office blood pressure measurement is simple and convenient, ambulatory blood pressure monitoring is especially useful for identifying patients with 'white coat' hypertension, in whom falsely raised office blood pressure recordings often lead to unnecessary return visits and additional treatment. Office and ambulatory blood pressure control was compared in 374 treated hypertensive patients in a single general practice, and the costs of performing annual ambulatory blood pressure measurements were compared with potential clinical savings. Ambulatory blood pressure monitoring detected 115 (31%) patients who fulfilled the British Hypertension Society target for ambulatory but not office blood pressure, i.e. white coat hypertension, and 21 patients apparently controlled by office but not ambulatory blood pressure criteria. In economic modelling, the capital, maintenance and user costs of ambulatory blood pressure monitoring (13,790 Pounds per year) were partly offset by fewer follow-up visits and second-line treatments in the group with white coat hypertension (there were modelled savings of 10,178 Pounds). Thus, ambulatory blood pressure monitoring identified a much larger number of treated hypertensive patients with adequate blood pressure control at an extra net cost of 3612 Pounds per year.


Effect of Na adsorption on the structural and electronic properties of Si(111)√3 × √3-Au surface.
Adsorption of ∼0.1 ML of Na onto the Si(111)√3 × √3-Au surface held at 300 °C has been found to induce pronounced changes in its structural and electronic properties. Domain wall networks, characteristic of the pristine surface, are removed completely, leading to the formation of a highly ordered homogeneous surface. The original atomic arrangement of the Si(111)√3 × √3-Au is preserved and Na atoms occupy T4 adsorption sites at the centers of surface Si trimers. Upon Na adsorption, a pronounced metallic S1 surface-state band develops. It is characterized by a large spin splitting (momentum splitting at the Fermi level Δk∥ = 0.027 Å(-1) and consequent energy splitting ΔEF = 110 meV), large electron filling (on the order of 0.5 electrons per √3 × √3 unit cell) and small effective electron mass of (0.028 ± 0.006)me. The natural consequence of the latter properties is a high surface conductivity of the Si(111)√3 × √3-(Au, Na) surface.


Weight maintenance and additional weight loss with liraglutide after low-calorie-diet-induced weight loss: the SCALE Maintenance randomized study.
Liraglutide, a once-daily human glucagon-like peptide-1 analog, induced clinically meaningful weight loss in a phase 2 study in obese individuals without diabetes. The present randomized phase 3 trial assessed the efficacy of liraglutide in maintaining weight loss achieved with a low-calorie diet (LCD). Obese/overweight participants (≥18 years, body mass index ≥30 kg m(-2) or ≥27 kg m(-2) with comorbidities) who lost ≥5% of initial weight during a LCD run-in were randomly assigned to liraglutide 3.0 mg per day or placebo (subcutaneous administration) for 56 weeks. Diet and exercise counseling were provided throughout the trial. Co-primary end points were percentage weight change from randomization, the proportion of participants that maintained the initial ≥5% weight loss, and the proportion that lost ≥5% of randomization weight (intention-to-treat analysis). ClinicalTrials.gov identifier: NCT00781937. Participants (n=422) lost a mean 6.0% (s.d. 0.9) of screening weight during run-in. From randomization to week 56, weight decreased an additional mean 6.2% (s.d. 7.3) with liraglutide and 0.2% (s.d. 7.0) with placebo (estimated difference -6.1% (95% class intervals -7.5 to -4.6), P<0.0001). More participants receiving liraglutide (81.4%) maintained the ≥5% run-in weight loss, compared with those receiving placebo (48.9%) (estimated odds ratio 4.8 (3.0; 7.7), P<0.0001), and 50.5% versus 21.8% of participants lost ≥5% of randomization weight (estimated odds ratio 3.9 (2.4; 6.1), P<0.0001). Liraglutide produced small but statistically significant improvements in several cardiometabolic risk factors compared with placebo. Gastrointestinal (GI) disorders were reported more frequently with liraglutide than placebo, but most events were transient, and mild or moderate in severity. Liraglutide, with diet and exercise, maintained weight loss achieved by caloric restriction and induced further weight loss over 56 weeks. Improvements in some cardiovascular disease-risk factors were also observed. Liraglutide, prescribed as 3.0 mg per day, holds promise for improving the maintenance of lost weight.


Influence of Rb/Cs Cation-Exchange on Inorganic Sn Halide Perovskites: From Chemical Structure to Physical Properties.
CsSnI3 is a potential lead-free inorganic perovskite for solar energy applications due to its nontoxicity and attractive optoelectronic properties. Despite these advantages, photovoltaic cells using CsSnI3 have not been successful to date, in part due to low stability. We demonstrate how gradual substitution of Rb for Cs influences the structural, thermodynamic, and electronic properties on the basis of first-principles density functional theory calculations. By examining the effect of the Rb:Cs ratio, we reveal a correlation between octahedral distortion and band gap, including spin-orbit coupling. We further highlight the cation-induced variation of the ionization potential (work function) and the importance of surface termination for tin-based halide perovskites for engineering high-performance solar cells.


Prospective comparative study of the evolution of probable Alzheimer's disease and Parkinson's disease dementia.
No previous comparison of test performance in probable Alzheimer's disease (pAD) and Parkinson's disease (PD) dementia has provided information about potential differences in the dementing process. This study compared the evolution of cognitive changes associated with these dementias. Generalized estimating equations (GEE) applied to regression analyses with repeated measures were used to evaluate cognitive changes over 1 to 3 years prior to the point when dementia was diagnosed in 40 matched pairs of patients with incident pAD and PD dementia. Both groups' performance declined on the Short Blessed, Selective Reminding Test (SRT; total recall, long-term retrieval, and delayed recall), Boston Naming Test, Category Fluency, and Similarities. The decline on naming and SRT delayed recall was more rapid in the PD dementia group, suggesting that these performance deficits emerge earlier in the development of pAD. The PD dementia group performed worse on Category Fluency throughout the follow-up period, suggesting either that dementia is overlaid on this preexisting performance deficit or that this type of executive deficit is an early manifestation of dementia in PD. The pAD group performed more poorly throughout the follow-up period on SRT delayed recognition, consistent with a pAD-specific encoding deficit. We conclude that while pAD and PD dementia are similar in many respects, differences in their evolution support previous observation of unique features in the 2 dementias and suggest different underlying pathologies.


Macular Fluid Reduces Reproducibility of Choroidal Thickness Measurements on Enhanced Depth Optical Coherence Tomography.
To determine if different types of retinal fluid in the central macula affect the reproducibility of choroidal thickness (CT) measurements on enhanced depth imaging optical coherence tomography (EDI-OCT). Retrospective reliability analysis. EDI-OCT images were obtained and the choroidal-scleral junction was analyzed through semiautomated segmentation. CT was measured at the fovea and averaged across the central 3-mm horizontal segment. Demographic data, central macular thickness, and type of fluid present were recorded. Intragrader and intergrader repeatability were assessed using the intraclass correlation coefficient (ICC) and coefficient of repeatability (CR). Of 124 eyes analyzed, 60 (48.4%) had diabetic macular edema, 32 (25.8%) had neovascular age-related macular degeneration, and 32 (25.8%) had other causes of fluid. Intergrader ICC (CR) was 0.95 (74.1 μm) and 0.96 (63.9 μm) for subfoveal and average CT, respectively. CR was similar across various causes of retinal fluid, but was worst for subretinal fluid compared to intraretinal or sub-retinal pigment epithelial fluid. CR also worsened with increasing choroidal thickness, but was not affected by retinal thickness. Intragrader repeatability was generally greater than intergrader values, and followed the same trend. The presence of macular fluid reduces CT measurement reproducibility, particularly in eyes with subretinal fluid and greater choroidal thickness. A difference of 74.1 μm in subfoveal CT or 63.9 μm in average CT may be necessary to detect true clinical change in eyes with macular fluid.


Conformation dependent monoclonal antibodies distinguish different replicating strains or conformers of prefibrillar Aβ oligomers.
Age-related neurodegenerative diseases share a number of important pathological features, such as accumulation of misfolded proteins as amyloid oligomers and fibrils. Recent evidence suggests that soluble amyloid oligomers and not the insoluble amyloid fibrils may represent the primary pathological species of protein aggregates. We have produced several monoclonal antibodies that specifically recognize prefibrillar oligomers and do not recognize amyloid fibrils, monomer or natively folded proteins. Like the polyclonal antisera, the individual monoclonals recognize generic epitopes that do not depend on a specific linear amino acid sequence, but they display distinct preferences for different subsets of prefibrillar oligomers. Immunological analysis of a number of different prefibrillar Aβ oligomer preparations show that structural polymorphisms exist in Aβ prefibrillar oligomers that can be distinguished on the basis of their reactivity with monoclonal antibodies. Western blot analysis demonstrates that the conformers defined by the monoclonal antibodies have distinct size distributions, indicating that oligomer structure varies with size. The different conformational types of Aβ prefibrillar oligomers can serve as they serve as templates for monomer addition, indicating that they seed the conversion of Aβ monomer into more prefibrillar oligomers of the same type. These results indicate that distinct structural variants or conformers of prefibrillar Aβ oligomers exist that are capable of seeding their own replication. These conformers may be analogous to different strains of prions.


Phenotyping EMT and MET cellular states in lung cancer patient liquid biopsies at a personalized level using mass cytometry.
Malignant pleural effusions (MPEs) can be utilized as liquid biopsy for phenotyping malignant cells and for precision immunotherapy, yet MPEs are inadequately studied at the single-cell proteomic level. Here we leverage mass cytometry to interrogate immune and epithelial cellular profiles of primary tumors and pleural effusions (PEs) from early and late-stage non-small cell lung cancer (NSCLC) patients, with the goal of assessing epithelial-mesenchymal transition (EMT) and mesenchymal-epithelial transition (MET) states in patient specimens. By using the EMT-MET reference map PHENOSTAMP, we observe a variety of EMT states in cytokeratin positive (CK+) cells, and report for the first time MET-enriched CK+ cells in MPEs. We show that these states may be relevant to disease stage and therapy response. Furthermore, we found that the fraction of CD33+ myeloid cells in PEs was positively correlated to the fraction of CK+ cells. Longitudinal analysis of MPEs drawn 2 months apart from a patient undergoing therapy, revealed that CK+ cells acquired heterogeneous EMT features during treatment. We present this work as a feasibility study that justifies deeper characterization of EMT and MET states in malignant cells found in PEs as a promising clinical platform to better evaluate disease progression and treatment response at a personalized level.


The double-stranded RNA binding protein RDE-4 can act cell autonomously during feeding RNAi in C. elegans.
Long double-stranded RNA (dsRNA) can silence genes of matching sequence upon ingestion in many invertebrates and is therefore being developed as a pesticide. Such feeding RNA interference (RNAi) is best understood in the worm Caenorhabditis elegans, where the dsRNA-binding protein RDE-4 initiates silencing by recruiting an endonuclease to process long dsRNA into short dsRNA. These short dsRNAs are thought to move between cells because muscle-specific rescue of rde-4 using repetitive transgenes enables silencing in other tissues. Here, we extend this observation using additional promoters, report an inhibitory effect of repetitive transgenes, and discover conditions for cell-autonomous silencing in animals with tissue-specific rescue of rde-4. While expression of rde-4(+) in intestine, hypodermis, or neurons using a repetitive transgene can enable silencing also in unrescued tissues, silencing can be inhibited wihin tissues that express a repetitive transgene. Single-copy transgenes that express rde-4(+) in body-wall muscles or hypodermis, however, enable silencing selectively in the rescued tissue but not in other tissues. These results suggest that silencing by the movement of short dsRNA between cells is not an obligatory feature of feeding RNAi in C. elegans. We speculate that similar control of dsRNA movement could modulate tissue-specific silencing by feeding RNAi in other invertebrates.


Extramedullary intradural spinal tumors: a pictorial review.
Defining the location of tumors and mass lesions of the spine in relation to the spinal cord and the dura is of the utmost importance as certain types of lesions tend to occur in certain locations. The differential diagnostic considerations will vary according to location of the mass lesion as will the treatment and prognosis of these various lesions. The category of extramedullary intradural masses includes a variety of lesions from meningiomas and nerve sheath tumors (neurofibromas, schwannomas) to less common tumors (hemangiopericytoma), metastases, benign tumors (lipoma, dermoid, epidermoid), inflammatory disorders (arachnoid adhesions, sarcoidosis), vascular lesions (spinal-dural arteriovenous fistula), and cystic lesions (perineural or Tarlov cysts). Characteristic magnetic resonance imaging findings are helpful for localization and characterization of these lesions before treatment, as well as for follow-up after treatment. We present a pictorial review of the various extramedullary intradural lesions of the spine, with pathologic correlation. We discuss imaging features that are typical for the various entities and describe various therapeutic options that are important considerations for surgical treatment of these lesions.


The caries process and its effect on the pulp: the science is changing and so is our understanding.
The understanding of the caries process and its effect on the pulp is presented in the context that caries does develop in various rates of progression. Early in the caries process, the pulp reflects changes within lesion activity. Thus, the early pulp response is reversible. Later, the rate of caries progression is reflected by the quality of the tertiary dentin. Slowly progressing lesions create tertiary dentin resembling normal tubular dentin. Rapidly progressing lesions lead to the production of atubular dentin or complete absence of tertiary dentin, as well as pulp necrosis and apical pathology. Finally, the nature of the untreated deep carious lesion is an ecosystem that might undergo significant changes. The untreated lesion is temporarily converted from an active and closed lesion environment into one that is open and slowly progressing. The analysis of untreated carious lesions has transformed the treatment philosophy of deep carious lesions.


The association of neighborhood-level social class and tobacco consumption with adverse lung cancer characteristics in Maryland.
Although both active tobacco use and passive tobacco exposure are well-established as being risk factors for lung cancer, it is challenging to measure tobacco-related exposures at the population level, while considering other factors (gender, race, socioeconomic status) that may modify the relationship between tobacco and lung cancer. Moreover, research to date has focused primarily on relationships between tobacco and endpoints of lung cancer incidence or mortality. Tobacco's role in disease progression, through association with important disease characteristics such as tumor histological type and grade, and stage of disease at diagnosis, has been less well examined. This research examines associations between area-level tobacco use and social class, as well as individual gender, race and age, and three adverse disease characteristics (tumor type, grade and stage) among incident cases of lung cancer reported to the Maryland Cancer Registry in 2000. Cases were geocoded by residential address. Multi-level logistic regression models included Census block group-level estimates of per capita tobacco spending, from Consumer Expenditure Survey data, and a 4-item social class index, from Census estimates of rates of high school graduation, employment, white collar occupation, and per capita income. Analyses of 3223 cases found no significant differences by race, however, results differed by gender. Lower block-group social class and higher tobacco spending were associated with squamous and small cell histological types and poorly differentiated or undifferentiated tumor grade. However, for later stage at diagnosis (SEER stages 2-7), both higher social class and greater tobacco spending were protective, especially for women, suggesting women in high tobacco use communities may benefit from early detection. Results support using area-level behavioral data as tools for identifying high risk communities suitable for more resource-intensive research or interventions. Findings also suggest that area-level social resources are consistent drivers of lung cancer disparities, and merit continued research attention.


The pharmacologic closure of the patent ductus arteriosus.
The ductus arteriosus is a fetal vessel that allows most of the blood leaving the right ventricle of the heart to bypass the lungs. Fetal patency of the ductus, and its spontaneous closure after birth, is the result of a balanced interaction of locally produced and circulating mediators (of which prostaglandins seem to be the most important), and the unique structure of the vessel wall. Persistent patency of the ductus occurs in almost 60% of very low birthweight infants. A significant left-to-right shunt through the ductus increases morbidity and mortality in premature infants. As prostaglandins play a major role in patency of the ductus, cyclooxygenase inhibitors are conventionally used to induce its closure. This chapter focuses on some of the basic mechanisms underlying ductal patency and the clinical attempts to diminish side effects associated with indomethacin, including the alternative use of ibuprofen.


Effects of Autologous Fat and ASCs on Swine Hypertrophic Burn Scars: A Multimodal Quantitative Analysis.
Hypertrophic scar formation is unpredictable and poorly understood, afflicting both the pediatric and adult populations. Treatment methods with conservative and invasive approaches have low rates of compliance and high rates of morbidity. The purpose of this study was to test a reproducible scar model and investigate a new technique of scar modification through the use of adipose- derived progenitor stromal cells (ASCs). Twenty thermal deep-partial thickness contact burns were created on the dorsum of three 8-week-old domestic swine and allowed to mature for 10 weeks. Scars were then injected with 2 cc saline, expanded autologous ASCs, or 2 cc fresh lipoaspirate and sampled at 2 week intervals up to 10 weeks postinjection. Volumetric analysis with a 3-D scanner, mechanical elasticity testing through negative pressure transduction, and standardized photography evaluation with Image J was performed. RNA sequencing was performed on scar tissue samples, cultured cells, and fresh lipoaspirate to determine relevant gene transcription regulation. Immunohistochemistry was used to verify expression level changes within the scars. Volumetric analysis demonstrates a reduction in average scar thickness at 6 weeks when injected with ASCs (-1.6 cc3) and autologous fat (-1.95 cc3) relative to controls (-0.121 cc3; P < 0.05). A decrease in overall tissue compliance is observed with fat or ASC injection when compared with unburned skin at 8 weeks (35.99/37.94 versus 49.36 mm Hg × mm; P < 0.01). RNA sequencing demonstrates altered regulation of fibroblast gene expression and a decreased inflammatory profile when scars are injected with autologous fat/ASCs over controls. Early results suggest that autologous fat and/or ASCs may improve healing of hypertrophic scarring by altering the cellular and structural components during wound remodeling up to 20 weeks after injury. This may have beneficial applications in early treatment of large or cosmetically sensitive immature burn scars.


Evaluation of separate renal function by means of 99mTc-aprotinin uptake test.
The possibility that relative kidney uptake of technetium-99m aprotinin (TcA) might be indicative of separate renal function was investigated in 89 patients who underwent both effective renal plasma flow (ERPFs) and glomerular filtration rate (GFR) determination. A reference group consisted of 27 healthy volunteers, studied only with TcA. The correlation with ERPFs (r = .73) was similar to that previously reported and confirmed. The correlation with GFR (r = .68) was better if a subgroup of renal units with TcA uptake lower than 16% (lower normal limit) was considered. Most likely, glomerular filtration is a limiting factor of the tubular uptake of TcA, and when GFR is reduced, both parameters decrease in the same manner, while if GFR is normal the two parameters are relatively independent. The correlation between TcA and GFR in 32 children was very similar to that found in adults. TcA uptake test seems to be a useful indicator of separate renal function, providing morphological information at, the same time.


Physiological Concentrations of Ascorbate Cannot Prevent the Potentially Damaging Reactions of Protein Radicals in Humans.
The principal initial biological targets of free radicals formed under conditions of oxidative stress are the proteins. The most common products of the interaction are carbon-centered alkyl radicals which react rapidly with oxygen to form peroxyl radicals and hydroperoxides. All these species are reactive, capable of propagating the free radical damage to enzymes, nucleic acids, lipids, and endogenous antioxidants, leading finally to the pathologies associated with oxidative stress. The best chance of preventing this chain of damage is in early repair of the protein radicals by antioxidants. Estimate of the effectiveness of the physiologically significant antioxidants requires knowledge of the antioxidant tissue concentrations and rate constants of their reaction with protein radicals. Previous studies by pulse radiolysis have shown that only ascorbate can repair the Trp and Tyr protein radicals before they form peroxyl radicals under physiological concentrations of oxygen. We have now extended this work to other protein C-centered radicals generated by hydroxyl radicals because these and many other free radicals formed under oxidative stress can produce secondary radicals on virtually any amino acid residue. Pulse radiolysis identified two classes of rate constants for reactions of protein radicals with ascorbate, a faster one in the range (9-60) × 107 M-1 s-1 and a slow one with a range of (0.5-2) × 107 M-1 s-1. These results show that ascorbate can prevent further reactions of protein radicals only in the few human tissues where its concentration exceeds about 2.5 mM.


Effects of oxypolygelatin and dextran 70 on hemostatic variables in dogs.
To evaluate and compare coagulation variables following the administration of oxypolygelatin and dextran 70 to clinically healthy dogs. Randomized cross-over experimental study. A total of eight healthy adult female Beagles aged 2-4 years old and weighing 11.8 +/- 2.7 kg. The dogs received a 15-minute intravenous (IV) infusion of 5 mL kg-1 oxypolygelatin or 10 mL kg-1 6% dextran 70. Before (PRE) and at 2, 5, and 24 hours after administration, packed cell volume (PCV), total solids concentration (TS), prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen concentration (FIB), platelet numbers (Plat), factor VIII coagulant activity (VIII:C), von Willebrand factor antigen concentration (vWf:Ag) and platelet function and buccal mucosal bleeding time (BMBT) were measured. Platelet function was assessed using aggregation and by measuring ATP release from aggregating platelets over 6 minutes, with 20, 10, and 5 micro m ADP and 5 and 10 micro g of collagen mL-1 as platelet activation agonists. All baseline values were within our normal ranges, except for one dog that had low vWf:Ag PRE values prior to both dextran and oxypolygelatin administration. Following dextran and oxypolygelatin administration, the PCV and TP were significantly (p < 0.05) decreased. Plat, FIB, and vWf:Ag decreased, while BMBT and VIII:C increased following dextran administration. Dextran also caused a significant decrease in platelet aggregation in response to ADP. Oxypolygelatin caused a significant decrease in vWf:Ag, Plat, and FIB compared to PRE values. The total amount of ATP released, standardized to platelet number, did not vary significantly for either group at any sampling time from PRE values. No significant changes from PRE values were noted at any time in either group for PT or APTT. At the doses administered, both dextran and oxypolygelatin can interfere with hemostatic variables in healthy dogs, but dextran's effect is more profound and prolonged when compared to oxypolygelatin. Oxypolygelatin causes fewer hemostatic abnormalities when compared to dextran, making it a superior colloid for administration at the doses tested.


Communicative signals during joint attention promote neural processes of infants and caregivers.
Communicative signals such as eye contact increase infants' brain activation to visual stimuli and promote joint attention. Our study assessed whether communicative signals during joint attention enhance infant-caregiver dyads' neural responses to objects, and their neural synchrony. To track mutual attention processes, we applied rhythmic visual stimulation (RVS), presenting images of objects to 12-month-old infants and their mothers (n = 37 dyads), while we recorded dyads' brain activity (i.e., steady-state visual evoked potentials, SSVEPs) with electroencephalography (EEG) hyperscanning. Within dyads, mothers either communicatively showed the images to their infant or watched the images without communicative engagement. Communicative cues increased infants' and mothers' SSVEPs at central-occipital-parietal, and central electrode sites, respectively. Infants showed significantly more gaze behaviour to images during communicative engagement. Dyadic neural synchrony (SSVEP amplitude envelope correlations, AECs) was not modulated by communicative cues. Taken together, maternal communicative cues in joint attention increase infants' neural responses to objects, and shape mothers' own attention processes. We show that communicative cues enhance cortical visual processing, thus play an essential role in social learning. Future studies need to elucidate the effect of communicative cues on neural synchrony during joint attention. Finally, our study introduces RVS to study infant-caregiver neural dynamics in social contexts.


PubCrawler: keeping up comfortably with PubMed and GenBank.
The free PubCrawler web service (http://www.pubcrawler.ie) has been operating for five years and so far has brought literature and sequence updates to over 22 000 users. It provides information on a personalized web page whenever new articles appear in PubMed or when new sequences are found in GenBank that are specific to customized queries. The server also acts as an automatic alerting system by sending out short notifications or emails with the latest updates as soon as they become available. A new output format and more flexibility for the email formatting help PubCrawler cope with increasing challenges arising from browser incompatibilities and mail filters, therefore making it suitable for a wide range of users.


k-space sampling optimization for ultrashort TE imaging of cortical bone: applications in radiation therapy planning and MR-based PET attenuation correction.
The ultrashort echo-time (UTE) sequence is a promising MR pulse sequence for imaging cortical bone which is otherwise difficult to image using conventional MR sequences and also poses strong attenuation for photons in radiation therapy and PET imaging. The authors report here a systematic characterization of cortical bone signal decay and a scanning time optimization strategy for the UTE sequence through k-space undersampling, which can result in up to a 75% reduction in acquisition time. Using the undersampled UTE imaging sequence, the authors also attempted to quantitatively investigate the MR properties of cortical bone in healthy volunteers, thus demonstrating the feasibility of using such a technique for generating bone-enhanced images which can be used for radiation therapy planning and attenuation correction with PET/MR. An angularly undersampled, radially encoded UTE sequence was used for scanning the brains of healthy volunteers. Quantitative MR characterization of tissue properties, including water fraction and R2(∗) = 1/T2(∗), was performed by analyzing the UTE images acquired at multiple echo times. The impact of different sampling rates was evaluated through systematic comparison of the MR image quality, bone-enhanced image quality, image noise, water fraction, and R2(∗) of cortical bone. A reduced angular sampling rate of the UTE trajectory achieves acquisition durations in proportion to the sampling rate and in as short as 25% of the time required for full sampling using a standard Cartesian acquisition, while preserving unique MR contrast within the skull at the cost of a minimal increase in noise level. The R2(∗) of human skull was measured as 0.2-0.3 ms(-1) depending on the specific region, which is more than ten times greater than the R2(∗) of soft tissue. The water fraction in human skull was measured to be 60%-80%, which is significantly less than the >90% water fraction in brain. High-quality, bone-enhanced images can be generated using a reduced sampled UTE sequence with no visible compromise in image quality and they preserved bone-to-air contrast with as low as a 25% sampling rate. This UTE strategy with angular undersampling preserves the image quality and contrast of cortical bone, while reducing the total scanning time by as much as 75%. The quantitative results of R2(∗) and the water fraction of skull based on Dixon analysis of UTE images acquired at multiple echo times provide guidance for the clinical adoption and further parameter optimization of the UTE sequence when used for radiation therapy and MR-based PET attenuation correction.


[Expression of peroxisome proliferator-activated receptor (PPAR) gamma in Helicobacter pylori-infected gastric epithelium].
Peroxisome proliferator-activated receptor gamma (PPAR gamma), a nuclear transcription factor, plays a critical role in the regulation of gene expression associated with inflammation and cancer. PPAR gamma is expressed in human gastric cancer as well as in colon cancer. Activation of PPAR gamma by ligand produces pro-apoptotic effect and ameliorate growing of cancer cells. Helicobacter pylori (H. pylori) is a main etiologic agent for gastric inflammation, and raises cell turnover in gastric epithelium. Longstanding infection with this organism is related with the development of non-cardiac gastric cancer. The aim of this study was to investigate the effect of H. pylori on the expression of PPAR gamma protein and mRNA in chronic gastritis. Gastric biopsy samples were taken from H. pylori infected (n=18) and non-infected (n=21) patients during endoscopic examination. PPAR gamma expressions were assessed by real time polymerase chain reaction and immunohistochemistry. PPAR gamma was localized to the nuclei of the foveolar epithelial cells in both infected and non-infected mucosa. PPAR gamma protein expression was higher in H. pylori infected patients than in non-infected patients (3.8+/-0.4 vs. 2.6+/-1.0, H. pylori infected and non-infected, respectively; p<0.05). However, PPAR gamma mRNA levels were not significantly different between the two groups (24+/-18 vs. 29+/-25, H. pylori infected and noninfected, respectively). PPAR gamma expression is increased in the gastric mucosa of H. pylori infected chronic gastritis, which suggests a certain role of PPAR gamma in the mucosal inflammatory reaction to H. pylori infection.


The Influence of Gut Dysbiosis in the Pathogenesis and Management of Ischemic Stroke.
Recent research on the gut microbiome has revealed the influence of gut microbiota (GM) on ischemic stroke pathogenesis and treatment outcomes. Alterations in the diversity, abundance, and functions of the gut microbiome, termed gut dysbiosis, results in dysregulated gut-brain signaling, which induces intestinal barrier changes, endotoxemia, systemic inflammation, and infection, affecting post-stroke outcomes. Gut-brain interactions are bidirectional, and the signals from the gut to the brain are mediated by microbially derived metabolites, such as trimethylamine N-oxide (TMAO) and short-chain fatty acids (SCFAs); bacterial components, such as lipopolysaccharide (LPS); immune cells, such as T helper cells; and bacterial translocation via hormonal, immune, and neural pathways. Ischemic stroke affects gut microbial composition via neural and hypothalamic-pituitary-adrenal (HPA) pathways, which can contribute to post-stroke outcomes. Experimental and clinical studies have demonstrated that the restoration of the gut microbiome usually improves stroke treatment outcomes by regulating metabolic, immune, and inflammatory responses via the gut-brain axis (GBA). Therefore, restoring healthy microbial ecology in the gut may be a key therapeutic target for the effective management and treatment of ischemic stroke.


Adjuvant radiotherapy and chemotherapy in breast cancer: 30 year follow-up of survival.
The long term outcome (more than 15 years) of adjuvant treatment in patients with primary operable breast cancer has rarely been examined. A randomised clinical trial of radiotherapy, chemotherapy (28 day cycles of cyclophosphamide, methotrexate and 5-fluorouracil) or both on women with primary operable breast cancer (n = 322) was followed-up for a median of 27 years. 260 (81%) patients died, 204 (78%) from breast cancer. Cancer specific survival (SE) at 10 years, 20 years and 30 years was 41 (3)%, 34 (3)% and 33 (3)% respectively. Presence of more than 3 involved lymph nodes increased cancer-specific mortality (HR 1.88, 95% CI 1.34-2.63) after adjustment for age, socio-economic deprivation and adjuvant treatment. Both age (HR 1.63, 95% CI 1.19-2.22) and involved lymph nodes (HR 1.59, 95% CI 1.17-2.14) were significant predictors of all-cause mortality after adjustment for other factors. There was no significant difference in all-cause or cancer-specific survival between patients in each of the 3 treatment arms. The present study highlights the long term impact of node positive disease but does not indicate that any regimen was associated with significantly better long-term survival.


US health care costs: the crushing burden.
This chapter provides an overview of health care costs in the United States, including trends, sources and uses of funds, employers' role, and factors driving costs. It also reviews what analysts believe are cost drivers especially compared to other countries that have significantly lower health care costs and, often, better health outcomes. Within the US, there are also important differences by geography, further demonstrating that higher US costs do not reflect higher quality and greater patient and physician satisfaction. In fact, the opposite is often the case.


Comprehensive genomic access to vector integration in clinical gene therapy.
Retroviral vectors have induced subtle clonal skewing in many gene therapy patients and severe clonal proliferation and leukemia in some of them, emphasizing the need for comprehensive integration site analyses to assess the biosafety and genomic pharmacokinetics of vectors and clonal fate of gene-modified cells in vivo. Integration site analyses such as linear amplification-mediated PCR (LAM-PCR) require a restriction digest generating unevenly small fragments of the genome. Here we show that each restriction motif allows for identification of only a fraction of all genomic integrants, hampering the understanding and prediction of biological consequences after vector insertion. We developed a model to define genomic access to the viral integration site that provides optimal restriction motif combinations and minimizes the percentage of nonaccessible insertion loci. We introduce a new nonrestrictive LAM-PCR approach that has superior capabilities for comprehensive unbiased integration site retrieval in preclinical and clinical samples independent of restriction motifs and amplification inefficiency.


Cellular heterogeneity identified by single-cell alkaline phosphatase (ALP) via a SERRS-microfluidic droplet platform.
Alkaline phosphatase (ALP) is a useful indicator for disease state diagnosis and clinical outcome. Investigation of ALP expression among cells is still challenging since ALP expression in a single cell is too low to be detectable. In our work, an ultrasensitive, high-throughput analytical method was applied for ALP determination in a single cell by using a surface-enhanced resonance Raman scattering (SERRS)-based microfluidic droplet technique. An ALP catalyzed substrate (5-bromo-4-chloro-3-indolyl phosphate, BCIP) was used to evaluate ALP activity in the cell within one droplet. When BCIP was incubated with cells, ALP can catalyze a hydrolysis reaction of colorless BCIP and oxidize the intermediate compound to form blue 5,5'-dibromo-4,4'-dichloro-1H,1H-[2,2']biindolylidene-3,3'-dione (BCI), which is a resonant Raman-active species. The encapsulation of BCI in droplets is favorable for detecting extremely low levels of molecules due to an accumulation effect along with reaction time. The ALP concentration as low as 1.0 × 10-15 M can be successfully detected in a uniform droplet. In addition, cellular heterogeneity profiled by ALP expression on single-cell resolution was monitored with this SERRS-based microfluidic droplet technique. Ultrasensitive determination of ALP secreted from individual cells can help us to understand cell-to-cell heterogeneity.


How is our self altered in psychiatric disorders? A neurophenomenal approach to psychopathological symptoms.
The self is central in our experience and has often been assumed to be necessary for any kind of consciousness in philosophy. Recent investigations in neuroscience demonstrate a particular set of regions such as the cortical midline regions to be associated with the processing of stimuli specifically related to the self as distinguished from those remaining unrelated to the self. Furthermore, findings show a close overlap between self-related activity and high levels of resting state activity in especially anterior midline regions. Interestingly, recent findings in psychiatric disorders such as depression and schizophrenia show resting state abnormalities in exactly these regions, that is in the cortical midline structures. Based on phenomenal and neural observations, I here suggest a neurophenomenal approach that directly links neuronal and phenomenal features (without sandwiching cognitive or sensorimotor functions) to psychopathological symptoms of self in depression and schizophrenia.


Longitudinal visual field variability and the ability to detect glaucoma progression in black and white individuals.
To investigate racial differences in the variability of longitudinal visual field testing in a 'real-world' clinical population, evaluate how these differences are influenced by socioeconomic status, and estimate the impact of differences in variability on the time to detect visual field progression. This retrospective observational cohort study used data from 1103 eyes from 751 White individuals and 428 eyes from 317 black individuals. Linear regression was performed on the standard automated perimetry mean deviation values for each eye over time. The SD of the residuals from the trend lines was calculated and used as a measure of variability for each eye. The association of race with the SD of the residuals was evaluated using a multivariable generalised estimating equation model with an interaction between race and zip code income. Computer simulations were used to estimate the time to detect visual field progression in the two racial groups. Black patients had larger visual field variability over time compared with white patients, even when adjusting for zip code level socioeconomic variables (SD of residuals for Black patients=1.53 dB (95% CI 1.43 to 1.64); for white patients=1.26 dB (95% CI 1.14 to 1.22); mean difference: 0.28 (95% CI 0.15 to 0.41); p<0.001). The difference in visual field variability between black and white patients was greater at lower levels of income and led to a delay in detection of glaucoma progression. Black patients had larger visual field variability compared with white patients. This relationship was strongly influenced by socioeconomic status and may partially explain racial disparities in glaucoma outcomes.


Echocardiographic and electrocardiographic examination of clinically healthy, conscious ferrets.
To assess the feasibility of performing electrocardiography (ECG) and echocardiography on conscious ferrets and to establish reference values for this species under these conditions. End-systolic and end-diastolic left ventricular (LV) diameters, LV and interventricular septal thicknesses were measured by M-mode echocardiography, while left atrial, aortic and pulmonary trunk diameters were determined by two-dimensional echocardiography in 46 ferrets. In some animals the maximum velocities of blood flow in the aorta and the pulmonary trunk were also measured by spectral Doppler. ECG examinations were performed by using limb leads both in right lateral recumbency and in the so-called "hanging" position. In the course of the ECG examinations (n=43), the occurrences, directions, durations and amplitudes of the P, QRS and T waves and the duration of the PR- and QT intervals were recorded for all six leads. ECG and echocardiography were successfully performed on non-sedated ferrets in 90 and 96% of the cases, respectively. Males were significantly heavier and had larger cardiac dimensions compared to females. Performance of both echocardiography and ECG are feasible techniques in conscious ferrets, and reference values are now available for future comparisons.


Parents' awareness and perception of children's eye diseases in Chennai, India.
To report parents' awareness and perception of eye diseases in their children. Thirty-five parents and 16 eye care practitioners either participated in in-depth interviews or focus group discussions. Data on parents' awareness and perception were collected using interview guides with unstructured questions. Data were transcribed, familiarized, and coded, and themes were generated. Redundancy was considered as the end point of data collection. The study results conveyed that parents were aware of common eye problems like refractive error, squint, and cataract, except for amblyopia, in their children. The causative factors for ocular diseases were not well understood by parents. The parents' perception was that eye problems can be treated with food, such as eggs or carrots, and exercises. Most of the parents perceived squint as a sign of good luck and spectacle correction as a social stigma. One of the prerequisites of health-seeking behavior is knowledge of disease and their symptoms, which seems to be lacking in parents of children. The findings of this study suggest that programs to increase awareness of the causative factors, spectacle wearing, and on the harmful effects of squint should be conducted.


Functional and Oncological Outcomes of Open Partial Laryngectomy vs. Transoral Laser Surgery in Supraglottic Larynx Cancer.
The aim of the presented study was to evaluate the outcomes of open partial laryngectomy (OPL) versus transoral laser surgery (TLS) in patients operated on for supraglottic laryngeal carcinoma based on functional parameters (duration of hospitalization, oral nutritional status and duration of transition to oral feeding, weaning status and duration after tracheotomy, and postoperative voice results) and oncological results (overall survival rate, disease-specific survival rate, recurrence, and presence of second primary tumors) in both groups. All laryngeal carcinoma patients who had undergone either OPL or TLS in the period from January 2012 to March 2017 in our center and were followed-up at least for 36 months were included in the study. Statistical analyses were carried out using the t-test and the Mann-Whitney U test to compare the means, and the Kaplan-Meier test for survival analysis. Fifty patients (44 males and 6 females) met the study criteria, of whom 31 had undergone OPL and 19 TLS. Patients that underwent TLS had less tracheotomy needs, needed shorter hospitalization periods, and transitioned to oral feeding earlier, compared to those that underwent OPL. There were no significant differences between the two groups based on oral feeding rates and voice outcomes. The impact of TLS and OPL on organ preservation in supraglottic laryngeal cancer were comparable. For local recurrences, repeated endolaryngeal laser surgeries and adjuvant treatments could be used in the TLS patient group. There were no significant differences between the two groups based on overall survival rate and disease specific survival rate. Although no significant differences were found in our study between the two surgical procedures in terms of oncological outcomes, TLS appeared to produce better functional outcomes in supraglottic laryngeal carcinoma than OPL.


[Soviet politics of breaking international war law. Representatives of medical professions among the victims of the Katyń crime].
In April 1940 the Soviet authorities murdered nearly 25 thousand Polish citizens belonging mainly to the intelligentsia. The crime is known as the Katyń Crime because Katyń was the first place where the victims had been discovered. Representatives of different medical specializations were one of the most numerous groups among the victims. Despite of the sixty-year period from their death the identity of many of them has not been found out. In the years 1994 - 1995 the author participated in exhumations made in the Katyń Woods. As an expert of the Government Board he was responsible for identifying, describing and cataloguing all objects found during the exhumation works. It was then when he started to gather information on medical doctors killed in 1940. The data verification has not been terminated yet as many of the data are incomplete and ambiguous. Publication of the partial and incomplete data is necessary to complete estimative information found in literature. The author considers that wider research works on the subject will deepen reflections on effects of the Katyń Crime.


A gene network database for the identification of key genes for diagnosis, prognosis, and treatment in sepsis.
Sepsis and sepsis-related diseases cause a high rate of mortality worldwide. The molecular and cellular mechanisms of sepsis are still unclear. We aim to identify key genes in sepsis and reveal potential disease mechanisms. Six sepsis-related blood transcriptome datasets were collected and analyzed by weighted gene co-expression network analysis (WGCNA). Functional annotation was performed in the gProfiler tool. DSigDB was used for drug signature enrichment analysis. The proportion of immune cells was estimated by the CIBERSORT tool. The relationships between modules, immune cells, and survival were identified by correlation analysis and survival analysis. A total of 37 stable co-expressed gene modules were identified. These modules were associated with the critical biology process in sepsis. Four modules can independently separate patients with long and short survival. Three modules can recurrently separate sepsis and normal patients with high accuracy. Some modules can separate bacterial pneumonia, influenza pneumonia, mixed bacterial and influenza A pneumonia, and non-infective systemic inflammatory response syndrome (SIRS). Drug signature analysis identified drugs associated with sepsis, such as testosterone, phytoestrogens, ibuprofen, urea, dichlorvos, potassium persulfate, and vitamin B12. Finally, a gene co-expression network database was constructed ( https://liqs.shinyapps.io/sepsis/ ). The recurrent modules in sepsis may facilitate disease diagnosis, prognosis, and treatment.


New functional aspects of cathepsin D and cathepsin E.
Cathepsin D (CD) and cathepsin E are representative lysosomal and nonlysosomal aspartic proteinases, respectively, and play an important role in the degradation of proteins, the generation of bioactive proteins, antigen processing, etc. Recenty, several lines of evidence have suggested the involvement of these two enzymes in the execution of neuronal death pathways induced by aging, transient forebrain ischemia, and excessive stimulation of glutamate receptors with excitotoxins. CD has also been shown to mediate apoptosis induced by various stimuli and p53-dependent tumor suppression. To gain more insight into in vivo functions of CD, mice deficient in this enzyme were generated. The mutant animals showed a progressive atrophy of the intestinal mucosa, a massive destruction of lymphoid organs, and a profound accumulation of ceroid lipofuscin, and developed a phenotype resembling neuronal ceroid lipofucinosis, suggesting that CD is essential for proteolysis of proteins regulating cell growth and tissue homeostasis. It has also been shown that CD molecules secreted from human prostate carcinoma cells are responsible for the generation of angiostatin, a potent endogenous inhibitor of angiogenesis, suggesting its contribution to the prevention of tumor growth and angiogenesis-dependent growth of metastases. Interestingly, pro-CD from human breast carcinoma cells showed a significantly lower angiostatin-generating activity than that from prostate carcinoma cells. Since deglycosylated CD molecules from both carcinoma cells showed a low angiostatin-generating activity, this discrepancy appeared to be attributed to the difference in the carbohydrate structures of CD molecules between the two cell types and to contribute to their potency to prevent tumor growth and metastases.


You can hide but you can't run: apparent competition, predator responses and the decline of Arctic ground squirrels in boreal forests of the southwest Yukon.
Throughout much of North America's boreal forest, the cyclical fluctuations of snowshoe hare populations (Lepus americanus) may cause other herbivores to become entrained in similar cycles. Alternating apparent competition via prey switching followed by positive indirect effects are the mechanisms behind this interaction. Our purpose is to document a change in the role of indirect interactions between sympatric populations of hares and arctic ground squirrels (Urocitellus parryii plesius), and to emphasize the influence of predation for controlling ground squirrel numbers. We used mark-recapture to estimate the population densities of both species over a 25-year period that covered two snowshoe hare cycles. We analysed the strength of association between snowshoe hare and ground squirrel numbers, and the changes to the seasonal and annual population growth rates of ground squirrels over time. A hyperbolic curve best describes the per capita rate of increase of ground squirrels relative to their population size, with a single stable equilibrium and a lower critical threshold below which populations drift to extinction. The crossing of this unstable boundary resulted in the subsequent uncoupling of ground squirrel and hare populations following the decline phase of their cycles in 1998. The implications are that this sustained Type II predator response led to the local extinction of ground squirrels. When few individuals are left in a colony, arctic ground squirrels may also have exhibited an Allee effect caused by the disruption of social signalling of approaching predators.


Gene expression profile of human lymphocytes exposed to (211)At alpha particles.
In this study, the Whole Human Genome 44K DNA microarray assay was used for the first time to obtain gene expression profiles in human peripheral blood lymphocytes 2 h after exposure (in suspension) to 6.78 MeV mean energy alpha particles from extracellular (211)At. Lymphocytes were exposed to fluences of 0.3-9.6 x 10(6) alpha particles/cm(2) [corresponding to mean absorbed alpha-particle doses (D(alpha)) of 0.05-1.60 Gy] over 30 min. Significantly modulated expression was identified in 338 early-response genes. Up-regulated expression was evident in 183 early-response genes, while the remaining 155 were down-regulated. Over half of the up-regulated genes and 40% of the down-regulated genes had a known biological process related primarily to cell growth and maintenance and cell communication. Genes associated with cell death were found only in the up-regulated genes and those with development only in the down-regulated genes. Eight selected early-response genes that displayed a sustained up- or down-regulation (CD36, HSPA2, MS4A6A, NFIL3, IL1F9, IRX5, RASL11B and SULT1B1) were further validated in alpha-particle-irradiated lymphocytes of two human individuals using the TaqMan(R) RT-qPCR technique. The results confirmed the observed microarray gene expression patterns. The expression modulation profiles of IL1F9, IRX5, RASL11B and SULT1B1 genes demonstrated similar trends in the two individuals studied. However, no significant linear correlation between increasing relative gene expression and the alpha-particle dose was evident. The results suggest the possibility that a panel of genes that react to alpha-particle radiation does exist and that they merit further study in a greater number of individuals to determine their possible value regarding alpha-particle biodosimetry.


Invasive glioblastoma cells acquire stemness and increased Akt activation.
Glioblastoma multiforme (GBM) is the most frequent and most aggressive brain tumor in adults. The dismal prognosis is due to postsurgery recurrences arising from escaped invasive tumor cells. The signaling pathways activated in invasive cells are under investigation, and models are currently designed in search for therapeutic targets. We developed here an in vivo model of human invasive GBM in mouse brain from a GBM cell line with moderate tumorigenicity that allowed simultaneous primary tumor growth and dispersal of tumor cells in the brain parenchyma. This strategy allowed for the first time the isolation and characterization of matched sets of tumor mass (Core) and invasive (Inv) cells. Both cell populations, but more markedly Inv cells, acquired stem cell markers, neurosphere renewal ability, and resistance to rapamycin-induced apoptosis relative to parental cells. The comparative phenotypic analysis between Inv and Core cells showed significantly increased tumorigenicity in vivo and increased invasion with decreased proliferation in vitro for Inv cells. Examination of a large array of signaling pathways revealed extracellular signal-regulated kinase (Erk) down-modulation and Akt activation in Inv cells and an opposite profile in Core cells. Akt activation correlated with the increased tumorigenicity, stemness, and invasiveness, whereas Erk activation correlated with the proliferation of the cells. These results underscore complementary roles of the Erk and Akt pathways for GBM proliferation and dispersal and raise important implications for a concurrent inhibitory therapy.


Connecting binuclear Pd(III) and mononuclear Pd(IV) chemistry by Pd-Pd bond cleavage.
Oxidation of binuclear Pd(II) complexes with PhICl(2) or PhI(OAc)(2) has previously been shown to afford binuclear Pd(III) complexes featuring a Pd-Pd bond. In contrast, oxidation of binuclear Pd(II) complexes with electrophilic trifluoromethylating ("CF(3)(+)") reagents has been reported to afford mononuclear Pd(IV) complexes. Herein, we report experimental and computational studies of the oxidation of a binuclear Pd(II) complex with "CF(3)(+)" reagents. These studies suggest that a mononuclear Pd(IV) complex is generated by an oxidation-fragmentation sequence proceeding via fragmentation of an initially formed, formally binuclear Pd(III), intermediate. The observation that binuclear Pd(III) and mononuclear Pd(IV) complexes are accessible in the same reactions offers an opportunity for understanding the role of nuclearity in both oxidation and subsequent C-X bond-forming reactions.


Isokinetic dynamometry of the knee extensors and flexors in Iranian healthy males and females.
This paper explores the gender-related bilateral differences of extensor and flexor torques of the knee joint at low and high angular velocities in Iranian healthy males and females. 70 healthy subjects (29 males (26.61±4.34 yrs and 41 females with average age of 23.07±3.70 yrs)) were participated in this study. Isokinetic peak torque values for knee extensors and flexors in concentric and eccentric contraction modes were measured and flexors and extensors strength ratios (HQR) computed among both dominant and non-dominant legs in lying position at 60 and 180°.s-1angular velocities. There was significant gender-velocity interactions detected for knee flexor to extensor strength ratios presenting that increasing velocity escaled this, ratios in females more than males (p<0.05). There was no gender- velocity-leg side interaction (p>0.05). Bilateral differences were found for eccentric flexor peak torques (p<0.05). By increasing velocity, peak torque values decreased and HQR was increased (p<0.05). Measurement procedures including test position is an important factor when interpreting genderrelated and bilateral differences of isokinetic knee strength ratios in healthy individuals.


Acinar cell carcinoma of the pancreas associated with subcutaneous panniculitis.
Acinar cell carcinoma is a rare pancreatic malignant tumor, which can be associated with a particular manifestation: pancreatic panniculitis. It presents with erythematous subcutaneous nodules located mainly on the legs. The skin lesions can precede, be concurrent with or follow the pancreatic illness. The pathogenesis is not fully understood, but it is believed to be associated with high levels of serum lipase produced by the neoplasm, causing fat necrosis in tissues. The treatment of pancreatic panniculitis is directed at the underlying pancreatic disease, which may result in regression of the skin lesions. We report a case of pancreatic acinar cell carcinoma associated with subcutaneous panniculitis together with a review of the English literature. A 79-year-old woman, with a 13-month history of acute pancreatitis, asthenia, weight loss, and the CT finding of a mass in the pancreatic head, presented complaining of multiple, migrant and painful subcutaneous nodules on her lower extremities. High serum lipase levels were also present. She underwent a pancreaticoduodenectomy, with subsequent regression of the skin lesions and normalization of her serum lipase levels. Histology revealed an acinar cell carcinoma of the pancreas. Liver metastases occurred six months after surgery, and the patient underwent chemotherapy with gemcitabine. She is still alive 14 months after the initial surgery. Although rare, the association between acinar cell carcinoma and pancreatic panniculitis has been described in the literature. In a patient with subcutaneous nodules, the presence of acinar cell carcinoma should be considered in order to prevent long delays in the diagnosis and treatment of this pancreatic malignancy.


[Relationships between humor expression and self-acceptance, aggression, and altruism].
The structure of humor expression was clarified and its relationships with aggression, altruism, and self-acceptance were examined. In study 1, college students (n = 216) responded to a scale with items about humor expression. An exploratory factor analysis indicated three types of humor expression: aggressive, self-disparaging, and playful humor expression. In study 2, 119 college students responded to items about (a) humor expression, (b) aggression, (c) altruism, and (d) self-acceptance. The results showed positive relationships between aggressive humor expression and aggression, self-disparaging humor expression and self-acceptance, and playful humor expression and altruism.


A novel pathway for arsenic elimination: human multidrug resistance protein 4 (MRP4/ABCC4) mediates cellular export of dimethylarsinic acid (DMAV) and the diglutathione conjugate of monomethylarsonous acid (MMAIII).
Hundreds of millions of people worldwide are exposed to unacceptable levels of arsenic in drinking water. This is a public health crisis because arsenic is a Group I (proven) human carcinogen. Human cells methylate arsenic to monomethylarsonous acid (MMA(III)), monomethylarsonic acid (MMA(V)), dimethylarsinous acid (DMA(III)), and dimethylarsinic acid (DMA(V)). Although the liver is the predominant site for arsenic methylation, elimination occurs mostly in urine. The protein(s) responsible for transport of arsenic from the liver (into blood), ultimately for urinary elimination, are unknown. Human multidrug resistance protein 1 (MRP1/ABCC1) and MRP2 (ABCC2) are established arsenic efflux pumps, but unlike the related MRP4 (ABCC4) are not present at the basolateral membrane of hepatocytes. MRP4 is also found at the apical membrane of renal proximal tubule cells, making it an ideal candidate for urinary arsenic elimination. In the current study, human MRP4 expressed in HEK293 cells reduced the cytotoxicity and cellular accumulation of arsenate, MMA(III), MMA(V), DMA(III), and DMA(V) while two other hepatic basolateral MRPs (MRP3 and MRP5) did not. Transport studies with MRP4-enriched membrane vesicles revealed that the diglutathione conjugate of MMA(III), monomethylarsenic diglutathione [MMA(GS)(2)], and DMA(V) were the transported species. MMA(GS)(2) and DMA(V) transport was osmotically sensitive, allosteric (Hill coefficients of 1.4 ± 0.2 and 2.9 ± 1.2, respectively), and high affinity (K0.5 of 0.70 ± 0.16 and 0.22 ± 0.15 μM, respectively). DMA(V) transport was pH-dependent, with highest affinity and capacity at pH 5.5. These results suggest that human MRP4 could be a major player in the elimination of arsenic.


Kelch-like homologue 9 mutation is associated with an early onset autosomal dominant distal myopathy.
Distal myopathies are a heterogeneous group of disorders characterized by progressive weakness and muscular atrophy, beginning in distal limb muscles and affecting proximal limb muscles at a later stage. We studied a large German kindred with 10 affected members. Weakness and atrophy of the anterior tibial muscles started between the ages of 8 and 16 years, followed by atrophy of intrinsic hand muscles. Progression was slow, and patients retained the ability to walk until the seventh decade. Serum creatinine kinase levels were increased in the range of 150-1400 U/l. Muscle biopsies showed myopathic changes, whereas immunohistochemistry showed normal expression of marker proteins for muscular dystrophies. Patients had reduced sensation with stocking-glove distribution in the distal limbs in later life. Nerve conduction studies revealed no evidence of neuropathy. Genome-wide linkage analysis in this family revealed a new locus for distal myopathy at 9p21.2-p22.3 (multipoint logarithm of the odds ratio=4.21). By positional cloning we found a heterozygous mutation L95F in the Kelch-like homologue 9 gene, encoding a bric-a-brac Kelch protein. Molecular modelling indicated that the mutation may interfere with the interaction of the bric-a-brac domain with Cullin 3. Coimmunoprecipitation experiments confirmed that the mutation reduces association with Cullin 3 in the Kelch-like homologue 9-Cullin 3-E3 ubiquitin ligase complex, which is involved in ubiquitin-dependent protein degradation. We identified a unique form of early onset autosomal dominant distal myopathy which is associated with a Kelch-like homologue 9 mutation and interferes with normal skeletal muscle through a novel pathogenetic mechanism.


Alteration of estrogen-regulated gene expression in human cells induced by the agricultural and horticultural herbicide glyphosate.
Gene expression is altered in mammalian cells (MCF-7 cells), by exposure to a variety of chemicals that mimic steroid hormones or interact with endocrine receptors or their co-factors. Among those populations chronically exposed to these endocrine disruptive chemicals are persons, and their families, who are employed in agriculture or horticulture, or who use agricultural/horticultural chemicals. Among the chemicals most commonly used, both commercially and in the home, is the herbicide glyphosate. Although glyphosate is commonly considered to be relatively non-toxic, we utilized in vitro DNA microarray analysis of this chemical to evaluate its capacity to alter the expression of a variety of genes in human cells. We selected a group of genes, determined by DNA microarray analysis to be dysregulated, and used quantitative real-time PCR to corroborate their altered states of expression. We discussed the reported function of those genes, with emphasis on altered physiological states that are capable of initiating adverse health effects that might be anticipated if gene expression were significantly altered in either adults or embryos exposed in utero.


Addressing chronic pain with Focused Acceptance and Commitment Therapy in integrated primary care: findings from a mixed methods pilot randomized controlled trial.
Over 100 million Americans have chronic pain and most obtain their treatment in primary care clinics. However, evidence-based behavioral treatments targeting pain-related disability are not typically provided in these settings. Therefore, this study sought to: 1) evaluate implementation of a brief evidence-based treatment, Focused Acceptance and Commitment Therapy (FACT-CP), delivered by an integrated behavioral health consultant (BHC) in primary care; and 2) preliminarily explore primary (self-reported physical disability) and secondary treatment outcomes (chronic pain acceptance and engagement in valued activities). This mixed-methods pilot randomized controlled trial included twenty-six participants with non-cancer chronic pain being treated in primary care (54% women; 46% Hispanic/Latino). Active participants completed a 30-min individual FACT-CP visit followed by 3 weekly 60-min group visits and a booster visit 2 months later. An enhanced treatment as usual (ETAU) control group received 4 handouts about pain management based in cognitive-behavioral science. Follow-up research visits occurred during and after treatment, at 12 weeks (booster visit), and at 6 months. Semi-structured interviews were conducted to collect qualitative data after the last research visit. General linear mixed regression models with repeated measures explored primary and secondary outcomes. The study design and FACT-CP intervention were feasible and acceptable. Quantitative analyses indicate at 6-month follow-up, self-reported physical disability significantly improved pre-post within the FACT-CP arm (d = 0.64); engagement in valued activities significantly improved within both the FACT-CP (d = 0.70) and ETAU arms (d = 0.51); and chronic pain acceptance was the only outcome significantly different between arms (d = 1.04), increased in the FACT-CP arm and decreased in the ETAU arm. Qualitative data analyses reflected that FACT-CP participants reported acquiring skills for learning to live with pain, consistent with increased chronic pain acceptance. Findings support that FACT-CP was acceptable for patients with chronic pain and feasible for delivery in a primary care setting by a BHC. Results provide preliminary evidence for improved physical functioning after FACT-CP treatment. A larger pragmatic trial is warranted, with a design based on data gathered in this pilot. clinicaltrials.gov, NCT04978961 (27/07/2021).


Bioburden Method Suitability: A Practical Solution to Screening for Inhibition.
ANSI/AAMI/ISO 11737-1:2018 addresses validation of the method for determination of bioburden. Clause 7 states that validation shall consist of four items, one of which is "assessment of test method suitability to demonstrate lack of inhibition of growth." Information must be provided, either through a rationale or via testing, showing that no inhibitory properties from the product exist in the specified bioburden test. However, the standard has no guidance on specific test methods for this particular purpose. Therefore, a simple bioburden suitability screening presented here could be used to quickly assess whether a product that is tested for bioburden has inhibitory substances that would be manifested in the bioburden test. The screening procedure uses a vegetative microorganism, Staphylococcus aureus, inoculated onto the product or into the extract solution, followed by the bioburden test procedure. A recovery of 70% or greater is recommended as acceptable for confirming the bioburden test Suitability.


Quantitative Single-Embryo Profile of Drosophila Genome Activation and the Dorsal-Ventral Patterning Network.
During embryonic development of Drosophila melanogaster, the maternal-to-zygotic transition (MZT) marks a significant and rapid turning point when zygotic transcription begins and control of development is transferred from maternally deposited transcripts. Characterizing the sequential activation of the genome during the MZT requires precise timing and a sensitive assay to measure changes in expression. We utilized the NanoString nCounter instrument, which directly counts messenger RNA transcripts without reverse transcription or amplification, to study >70 genes expressed along the dorsal-ventral (DV) axis of early Drosophila embryos, dividing the MZT into 10 time points. Transcripts were quantified for every gene studied at all time points, providing the first dataset of absolute numbers of transcripts during Drosophila development. We found that gene expression changes quickly during the MZT, with early nuclear cycle 14 (NC14) the most dynamic time for the embryo. twist is one of the most abundant genes in the entire embryo and we use mutants to quantitatively demonstrate how it cooperates with Dorsal to activate transcription and is responsible for some of the rapid changes in transcription observed during early NC14. We also uncovered elements within the gene regulatory network that maintain precise transcript levels for sets of genes that are spatiotemporally cotranscribed within the presumptive mesoderm or dorsal ectoderm. Using these new data, we show that a fine-scale, quantitative analysis of temporal gene expression can provide new insights into developmental biology by uncovering trends in gene networks, including coregulation of target genes and specific temporal input by transcription factors.


Phase II study of docetaxel weekly in combination with carboplatin every 3 weeks as first-line chemotherapy in stage IIB to stage IV epithelial ovarian cancer.
The purpose of this study was to assess the response rate, toxicity, progression-free survival, and overall survival in a series of patients with advanced-stage ovarian carcinoma treated with a first-line weekly docetaxel and 3 weekly carboplatin regimen. All eligible patients were treated with intravenous docetaxel (30 mg/m) on days 1, 8, and 15, and carboplatin (area under the curve, 5) on day 1; every 21 days for at least 6 cycles. One hundred six patients received at least one cycle of primary chemotherapy (median, 6.0; range, 1-9), and they were evaluable for toxicity assessment. Eighty-five patients had evaluable (measurable) disease and received at least 3 courses of chemotherapy and were evaluable for clinical response rate. The overall response rate was 78.8% (95% confidence interval, 70.1%-87.5%), and the biochemical response 92.8% (95% confidence interval, 87.2%-98.4%). The median progression-free survival was 12.0 months and the median overall survival was 35.3 months. Thirty-six patients (34.0%) experienced grades 3 and 4 neutropenia, which resulted in the removal of 3 patients. Six patients (5.7%) experienced grades 3 or 4 thrombocytopenia. No patients experienced grade 3 to grade 4 sensory neuropathy. Epiphora, nail changes, and fatigue were frequently recorded nonhematologic adverse effects. The tolerable hematologic toxicity (no need for colony-stimulating factors) and the low rate of neurotoxicity (only grades 1-2) and response rates in line with the standard 3-week paclitaxel-carboplatin regimen for advanced primary ovarian carcinoma after suboptimal cytoreductive surgery make this regimen an interesting alternative in selected patients.


Japanese responses to social change--making the strange familiar.
Understanding the concept of a "sense of coherence" can be useful in trying to account for successful and healthy adaptations to situations of social change and migration. Certain fundamental dynamics of social life in Japan contribute to a sense of coherence in the lives of Japanese today. Analysts of modern Japan have noted that it has not become a replica of western societies in that primary social groups take precedence over individual needs and autonomy. Moreover, although the Japanese personality may be characterized as multilayered, one dominant aspect is the tendency to suppress negative feelings-towards intimates, family and those in authority. This tendency has implications for the patient-physician relationship, which tends to be ritualized to protect a patient's innermost feelings. Because there has never been a split between mind and body in Japanese thinking there is no concept of mental health that is separate from physical health; rather, patients and physicians readily accept that illness is an expression of stress on the social level. As a corollary, the responsibility for healing is felt to be in the hands of patients and their families, with physicians playing the part of skilled and sympathetic technicians. Ascribing the origins of a cultural identity risks stereotyping, but understanding the reasons for the continuity of certain values in Japanese immigrants will enable physicians to use these values to advantage, and explains the healthy adaptation of these immigrants to the disruption of migration.


Safety and immunogenicity of a bivalent Haemophilus influenzae type b/hepatitis B vaccine in healthy infants. Hib-HB Vaccine Study Group.
To assess the safety, tolerability and immunogenicity of COMVAX, a liquid, bivalent Haemophilus influenzae type b-hepatitis B vaccine, containing the polyribosylribitol phosphate (PRP)-Neisseria meningitidis outer membrane protein complex conjugate used in the Hib vaccine, PedvaxHIB, and the yeast-derived hepatitis B surface antigen (HBsAg) used in the HB vaccine, RECOMBIVAX HB. Eight hundred eighty-two healthy infants, approximately 2 months of age, were enrolled in an open, multicenter (n = 11) clinical trial and randomized to receive either COMVAX (7.5 micrograms of PRP/5 micrograms of HBsAg in 0.5 ml) or concurrent injections of the liquid formulation of PedvaxHIB (P) (7.5 micrograms of PRP in 0.5 ml) and RECOMBIVAX HB (R) (5 micrograms of HBsAg in 0.5 ml) at 2, 4 and 12 or 15 months of age. Safety and tolerability were monitored after each injection. The serum concentrations of anti-PRP and anti-HBs were determined at the time of each vaccination, 2 months after the second vaccination and 1 month after the third vaccination. COMVAX was well-tolerated and proved to be immunologically comparable with a series of concomitant P+R injections. There were no serious adverse experiences attributable to the study vaccines. The most commonly reported nonserious adverse experiences were all events prelisted on diary cards given to parents. These included generally mild and transient signs of inflammation at the injection site (pain/ soreness, erythema, swelling/induration), somnolence and irritability. Because children are at peak risk of invasive Hib disease during the first year of life, 6 months of age (2 months after the second dose of vaccine) was designated the time of primary interest with regard to the development of anti-PRP. At that time 94.8% of the infants given COMVAX had > 0.15 microgram/ml of anti-PRP and 72.4% had > 1.0 microgram/ ml, with a geometric mean concentration (GMC) of 2.5 micrograms/ml, compared with 95.2%, 76.3% and 2.8 micrograms/ml, respectively, in recipients of P+R. The third injection given at 12 or 15 months of age induced a secondary rise in antibody. The proportions with > 0.15 microgram/ml and > 1.0 microgram/ml of anti-PRP increased to 99.3 and 92.6%, respectively, and the GMC rose to 9.5 micrograms/ml among COMVAX recipients, compared with 98.9%, 92.3% and 10.2 micrograms/ml in children given concurrent injections of P+R. In contrast to Hib few infants in countries with low endemicity of HBV infection are at near term risk of exposure to virus. Consequently the anti-HBs response after the last dose of vaccine was designated the outcome of primary interest. At 13 to 16 months of age (1 month after the third dose of vaccine) 98.4% of children given COMVAX had a protective anti-HBs concentration of > or = 10 mIU/ml with a GMC of 4468 mIU/ml, compared with 100% and a GMC of 6944 mIU/ml among children given P+R. COMVAX is well-tolerated by healthy infants and can induce immunity against invasive Hib disease and HBV infection using only three injections compared with six injections if separate courses of monovalent PedvaxHIB and RECOMBIVAX HB are given.


Patterns of MHC-dependent sexual selection in a free-living population of sheep.
The MHC is one of the most polymorphic gene clusters in vertebrates and play an essential role in adaptive immunity. Apart from pathogen-mediated selection, sexual selection can also contribute to the maintenance of MHC diversity. MHC-dependent sexual selection could occur via several mechanisms but at present there is no consensus as to which of these mechanisms are involved and their importance. Previous studies have often suffered from limited genetic and behavioural data and small sample size, and were rarely able to examine all the mechanisms together, determine whether signatures of MHC-based non-random mating are independent of genomic effects or differentiate whether MHC-dependent sexual selection takes place at the pre- or post-copulatory stage. In this study, we use Monte Carlo simulation to investigate evidence for non-random MHC-dependent mating patterns by all three mechanisms in a free-living population of Soay sheep. Using 1710 sheep diplotyped at the MHC class IIa region and genome-wide SNPs, together with field observations of consorts, we found sexual selection against a particular haplotype in males at the pre-copulatory stage and sexual selection against female MHC heterozygosity during the rut. We also found MHC-dependent disassortative mating at the post-copulatory stage, along with strong evidence of inbreeding avoidance at both stages. However, results from generalized linear mixed models suggest that the pattern of MHC-dependent disassortative mating could be a by-product of inbreeding avoidance. Our results therefore suggest that while multiple apparent mechanisms of non-random mating with respect to the MHC may occur, some of them have alternative explanations.


In situ investigations on organic foam films using neutron and synchrotron radiation.
We report on small-angle neutron scattering (SANS) and X-ray scattering (SAXS) investigations of foam films stabilized by sodium dodecyl sulfate. Previous measurements on dry foams (Axelos, M. A. V.; Boue, B. Langmuir 2003, 19, 6598) have shown the presence of spikes in the two-dimensional scattering data which suggest that the incident beam is reflected on some film surfaces. The latter interpretation is confirmed by new neutron studies performed on ordered ("bamboo") foams which allow selection of single films. In the first case, we show that the spikes of the scattered intensity can be obtained by reflection on two parts of the foam, namely, the films and the Plateau borders. With synchrotron radiation, first observations of distinct interference fringes have allowed an accurate determination of the film thickness. A comparison with X-ray and neutron data is made, opening a general discussion about the capabilities of small-angle scattering techniques for studying the microscopic properties of foam films.


The multifaceted function of BAK1/SERK3: plant immunity to pathogens and responses to insect herbivores.
Almost a decade ago BRI1-associated kinase 1 (BAK1) was identified as a co-receptor of brassinosteroid (BR) insensitive 1 (BRI1), the receptor for BRs, which plays an essential role in transducing BR signaling to regulate plant development. BAK1 is also critical in resistance to various pathogens. BAK1 rapidly binds to certain receptors for pathogen/microbeassociated molecular patterns (PAMPs/MAMPs) after the perception of pathogen elicitors and is required for the full elicitation of pathogen-induced defense responses, such as the activation of the mitogen-activated protein kinase 6 (MPK6) and production of reactive oxygen species. Thus, BAK1 functions in both BR signaling and PAMP-triggered immunity (PTI). Recently BAK1 was also found to play an important role in mediating defense responses against an insect herbivore (Manduca sexta) of Nicotiana attenuata. In this interaction, BAK1 positively modulates wound- or herbivore feeding induced accumulation of jasmonic acid (JA) and JA-isoleucine (JA-Ile). This mini-review summarizes recent advances in our understanding of the functions of BAK1 in resistance to pathogens and herbivores.


Nisin M: a Bioengineered Nisin A Variant That Retains Full Induction Capacity but Has Significantly Reduced Antimicrobial Activity.
Nisin A is a potent antimicrobial with potential as an alternative to traditional antibiotics, and a number of genetically modified variants have been created that target clinically relevant pathogens. In addition to antimicrobial activity, nisin autoregulates its own production via a signal transduction pathway, a property that has been exploited in a protein expression system termed the nisin-controlled gene expression (NICE) system. Although NICE has become one of the most popular protein expression systems, one drawback is that the inducer peptide, nisin A, also has inhibitory activity. It has already been demonstrated that the N-terminal region of nisin A contributes to antimicrobial activity and signal transduction properties; therefore, we conducted bioengineering of nisin at positions Pro9 and Gly10 within ring B to produce a bank of variants that could potentially be used as alternative induction peptides. One variant, designated nisin M, has threonines at positions 9 and 10 and retains induction capacity comparable to that of wild-type nisin A, while most of the antimicrobial activity is abolished. Further analysis confirmed that nisin M produces a mix of peptides as a result of different degrees of dehydration of the two threonines. We show that nisin M exhibits potential as a more suitable alternative to nisin A for the expression of proteins that may be difficult to express or for production of proteins in strains that are sensitive to wild-type nisin. Moreover, it may address the increasing demand by industry for optimization of peptide fermentations to increase yields or production rates.IMPORTANCE This study describes the generation of a nisin variant with superior characteristics for use in the NICE protein expression system. The variant, termed nisin M, retains an induction capacity comparable to that of wild-type nisin A but exhibits significantly reduced antimicrobial activity and can therefore be used at concentrations that are normally toxic to the expression host.


Reconstruction of Ewing Sarcoma Developmental Context from Mass-Scale Transcriptomics Reveals Characteristics of EWSR1-FLI1 Permissibility.
Ewing sarcoma is an aggressive pediatric cancer of enigmatic cellular origins typically resulting from a single translocation event t (11; 22) (q24; q12). The resulting fusion gene, EWSR1-FLI1, is toxic or unstable in most primary tissues. Consequently, attempts to model Ewing sarcomagenesis have proven unsuccessful thus far, highlighting the need to identify the cellular features which permit stable EWSR1-FLI1 expression. By re-analyzing publicly available RNA-Sequencing data with manifold learning techniques, we uncovered a group of Ewing-like tissues belonging to a developmental trajectory between pluripotent, neuroectodermal, and mesodermal cell states. Furthermore, we demonstrated that EWSR1-FLI1 expression levels control the activation of these developmental trajectories within Ewing sarcoma cells. Subsequent analysis and experimental validation demonstrated that the capability to resolve R-loops and mitigate replication stress are probable prerequisites for stable EWSR1-FLI1 expression in primary tissues. Taken together, our results demonstrate how EWSR1-FLI1 hijacks developmental gene programs and advances our understanding of Ewing sarcomagenesis.


Effect of the E200K mutation on prion protein metabolism. Comparative study of a cell model and human brain.
The hallmark of prion diseases is the cerebral accumulation of a conformationally altered isoform (PrP(Sc)) of a normal cellular protein, the prion protein (PrP(C)). In the inherited form, mutations in the prion protein gene are thought to cause the disease by altering the metabolism of the mutant PrP (PrP(M)) engendering its conversion into PrP(Sc). We used a cell model to study biosynthesis and processing of PrP(M) carrying the glutamic acid to lysine substitution at residue 200 (E200K), which is linked to the most common inherited human prion disease. PrP(M) contained an aberrant glycan at residue 197 and generated an increased quantity of truncated fragments. In addition, PrP(M) showed impaired transport of the unglycosylated isoform to the cell surface. Similar changes were found in the PrP isolated from brains of patients affected by the E200K variant of Creutzfeldt-Jakob disease. Although the cellular PrP(M) displayed some characteristics of PrP(Sc), the PrP(Sc) found in the E200K brains was quantitatively and qualitatively different. We propose that the E200K mutation cause the same metabolic changes of PrP(M) in the cell model and in the brain. However, in the brain, PrP(M) undergoes additional modifications, by an age-dependent mechanism that leads to the formation of PrP(Sc) and the development of the disease.


Optimal Extraction and Deproteinization Method for Mannoprotein Purification from Kluyveromyces marxianus.
Mannoproteins, mannose-glycosylated proteins, play an important role in biological processes and have various applications in industries. Several methods have been already used for the extraction of mannoproteins from yeast cell-wall. The aim of this study was to evaluate the extraction and deproteinization of mannan oligosaccharide from the Kluyveromyces (K.) marxianus mannoprotein. To acquire crude mannan oligosaccharides, K. marxianus mannoproteins were deproteinized by the Sevage, trichloroacetic acid, and hydrochloric acid (HCL) methods. Total nitrogen, crude protein content, fat, carbohydrate and ash content were measured according to the monograph prepared by the meeting of the Joint FAO/WHO Expert Committee and standard. Mannan oligosaccharide loss, percentage of deproteinization, and chemical composition of the product were assessed to check the proficiency of different methods. Highly purified (95.4%) mannan oligosaccharide with the highest deproteinization (97.33 ± 0.4%) and mannan oligosaccharide loss (25.1 ± 0.6%) were obtained following HCl method. HCl, was the most appropriate deproteinization method for the removal of impurities. This preliminary data will support future studies to design scale-up procedures.