Heterochromatin protein 1 is required for correct chromosome segregation in Drosophila embryos.
Heterochromatin protein 1 is associated with centromeric heterochromatin in Drosophila, mice, and humans. Loss of function mutations in the gene encoding heterochromatin protein 1 in Drosophila, Suppressor of variegation2-5, decrease the mosaic repression observed for euchromatic genes that have been juxtaposed to centromeric heterochromatin. These heterochromatin protein 1 mutations not only suppress this position-effect variegation, but also cause recessive embryonic lethality. In this study, we analyze the latter phenotype in the hope of gaining insight into heterochromatin function. In our analyses of four alleles of Suppressor of variegation2-5, the lethality was found to be associated with defects in chromosome morphology and segregation. While some of these defects are seen throughout embryonic development, both the frequency and severity of the defects are greatest between cycles 10 and 14 when zygotic transcription of the Suppressor of variegation2-5 gene apparently begins. By this time in development, heterochromatin protein 1 levels are diminished by four-fold in a quarter of the embryos produced by parents that are both heterozygous for a null allele (Suppressor of variegation2-5(05)). In a live analysis of the phenotype, we find prophase to be lengthened by more than two-fold in Suppressor of variegation2-5(05) mutant embryos with subsequent defects in chromosome segregation. The elongated prophase suggests that the segregation phenotype is a consequence of defects in events that occur during prophase, either in chromosome condensation or kinetochore assembly or function. Immunostaining with an antibody against a centromerespecific antigen indicates that the kinetochores of most chromosomes are functional. The immunostaining results are more consistent with defects in chromosome condensation being responsible for the segregation phenotype.


Disseminated histoplasma and CMV infection presenting as subacute intestinal obstruction in an immunocompromised patient.
Histoplasma in patients with impaired cellular immunity can disseminate to various organs and is known as progressive disseminated histoplasmosis. Similarly cytomegalovirus (CMV) is the most common opportunistic pathogen in an immunocompromised host. The authors report an older male with symptoms and radiological evidence of subacute intestinal obstruction. The patient had concerns for compromised immune system as he was on chronic prednisone and methotrexate therapy. Follow-up colonoscopy revealed a stricture in the proximal ascending colon. Biopsy of the stricture revealed ulcer with granulomatous inflammation including well-formed granulomas and an infiltrate of histiocytes within the lamina propria. Special stains on the specimen showed fungal structures consistent with Histoplasma capsulatum. Immunochemistry showed presence of CMV in the tissue. The patient had good response to antimicrobial therapy and did not have progression of intestinal obstruction. This case highlights the need to consider infectious pathology in immunocompromised patients presenting with obstructive symptoms.


Acinetobacter baumannii strains isolated from patients in intensive care units in Goiânia, Brazil: Molecular and drug susceptibility profiles.
Resistance to antimicrobial agents is increasing worldwide and imposes significant life-threatening risks to several different populations, especially those in intensive care units (ICUs). Bacteria can quickly develop or acquire resistance to antimicrobial drugs, and combined with their intrinsic potential to cause disease in humans, these bacteria can become deadly. Among Gram-negative bacteria, Acinetobacter baumannii is notorious as a frequent opportunistic pathogen associated with critically ill patients, and understanding the genetic basis of A. baumannii resistance to beta-lactams among patients in ICUs will result in better protocols to prevent the development of resistance as well as improved treatment regimens. In this study, we assessed 1333 patients in five ICUs, 56 of whom developed A. baumannii infections. Most of the A. baumannii isolates were resistant to beta-lactam antimicrobial drugs, specifically, 3rd- and 4th-generation cephalosporins and carbapenems, and 91.1% of the isolates were multi-drug resistant (MDR). The most frequent OXA gene present was OXA-23 (55.1%), which is significantly associated with MDR strains. Most of the A. baumannii isolates (76.8%) were capable of forming a biofilm. The antimicrobial drug classes that were effective against most of these isolates were polymyxins and tigecycline. The molecular profile of the isolates allowed detection of 12 different clusters comprising 2 to 8 isolates each. In conclusion, our data indicate a high incidence of resistance to carbapenems as well as MDR strains among the observed A. baumannii isolates, most of which exhibited a high prevalence of OXA-23 gene expression. Only a few selective drugs were effective, reinforcing the notion that bacterial resistance is an emerging problem that should be prioritized in every healthcare facility.


Mechanism of impaired baroreflex control in prehypertensive Dahl salt-sensitive rats.
Dahl salt-sensitive rats fed a low salt diet demonstrate functional impairment in baroreflex control of cardiovascular function prior to the development of hypertension. The purpose of this study was to identify the mechanism(s) responsible for impaired baroreflex control in prehypertensive Dahl salt-sensitive rats. To examine the central and efferent portions of the baroreflex arc, we electrically stimulated the aortic depressor nerve of urethane anesthetized Dahl salt-sensitive and Dahl salt-resistant rats while recording arterial pressure, heart rate, and sympathetic nerve activity. Aortic nerve stimulation produced equivalent responses in both groups, indicating that central and efferent mechanisms could not account for functional baroreflex impairment. In separate groups of Dahl rats, multifiber afferent aortic baroreceptor discharge was recorded while arterial pressure was increased with phenylephrine. Prehypertensive Dahl salt-sensitive rats demonstrated significantly less baroreceptor activation than Dahl salt-resistant rats, indicating that impaired baroreceptor discharge is the mechanism responsible for baroreflex abnormalities. Aortic arch distensibility was not different between Dahl strains, suggesting that impaired baroreceptor discharge in Dahl salt-sensitive rats is probably not due to abnormalities in vessel wall distensibility. Since arterial pressure of both Dahl strains was in the normotensive range and did not differ significantly between Dahl salt-sensitive and salt-resistant rats, baroreceptor impairment appears to be a primary defect in Dahl salt-sensitive rats, rather than being secondary to hypertension. We speculate that this impairment in baroreceptor function in Dahl salt-sensitive rats prior to any elevation in arterial pressure may contribute to the Dahl salt-sensitive rat's genetic propensity to develop hypertension.


Recognition of face-name associations after errorless and errorful learning: an fMRI study.
Errorless learning has advantages over errorful learning. The erroneous items produced during errorful learning compete with correct items at retrieval resulting in decreased memory performance. This interference is associated with an increased demand on executive monitoring processes. Event-related functional magnetic resonance imaging (fMRI) was used to contrast errorless and errorful learning. Learning mode was manipulated by the number of distractors during learning of face-name associations: in errorless learning only the correct name was introduced. During errorful learning either one incorrect name or two incorrect names were additionally introduced in order to modulate the interference in recognition. The behavioural results showed an enhanced memory performance after errorless learning. The veridicality of recognition of the face-name associations was reflected in a left lateralized fronto-temporal-parietal network. The different learning modes were associated with modulations in left prefrontal and parietal regions. Errorless learning enhances memory performance as compared to errorful learning and underpins the known advantages for errorless learning. During memory retrieval different networks are engaged for specific purposes: Recognition of face-name associations engaged a lateralized fronto-temporal-parietal network and executive monitoring processes of memory engaged the left prefrontal and parietal regions.


ProtFus: A Comprehensive Method Characterizing Protein-Protein Interactions of Fusion Proteins.
Tailored therapy aims to cure cancer patients effectively and safely, based on the complex interactions between patients' genomic features, disease pathology and drug metabolism. Thus, the continual increase in scientific literature drives the need for efficient methods of data mining to improve the extraction of useful information from texts based on patients' genomic features. An important application of text mining to tailored therapy in cancer encompasses the use of mutations and cancer fusion genes as moieties that change patients' cellular networks to develop cancer, and also affect drug metabolism. Fusion proteins, which are derived from the slippage of two parental genes, are produced in cancer by chromosomal aberrations and trans-splicing. Given that the two parental proteins for predicted fusion proteins are known, we used our previously developed method for identifying chimeric protein-protein interactions (ChiPPIs) associated with the fusion proteins. Here, we present a validation approach that receives fusion proteins of interest, predicts their cellular network alterations by ChiPPI and validates them by our new method, ProtFus, using an online literature search. This process resulted in a set of 358 fusion proteins and their corresponding protein interactions, as a training set for a Naïve Bayes classifier, to identify predicted fusion proteins that have reliable evidence in the literature and that were confirmed experimentally. Next, for a test group of 1817 fusion proteins, we were able to identify from the literature 2908 PPIs in total, across 18 cancer types. The described method, ProtFus, can be used for screening the literature to identify unique cases of fusion proteins and their PPIs, as means of studying alterations of protein networks in cancers. Availability: http://protfus.md.biu.ac.il/.


N-(5-indanylsulfonyl)-N'-(4-chlorophenyl)urea, a novel agent equally cytotoxic to nonproliferating human colon adenocarcinoma cells.
Diarylsulfonylureas have been shown to have therapeutic activity against rodent and human tumor models, notably causing regressions in some lines of human colon adenocarcinomas in mice. At present the mechanism of cytotoxicity is unknown, although preliminary data implicate mitochondria as a potential site of action. In this study, the cytotoxicity of the diarylsulfonylurea N-(5-indanylsulfonyl)-N'-(4-chlorophenyl)urea (ISCU) has been examined in GC3/c1 human colon adenocarcinoma cells. At cytotoxic concentrations of ISCU, in the presence of albumin as a drug binding species, there was only slight inhibition of [3H]thymidine and [3H]uridine incorporation at concentrations of ISCU up to 140 micrograms/ml and no inhibition of synthesis of protein as determined by incorporation of L-leucine. In the absence of albumin, incorporation of [3H]thymidine into DNA or [3H]uridine into RNA was inhibited at greater than 70 micrograms/ml and 140 micrograms/ml, respectively. As ISCU is highly bound to serum albumin (greater than 99%), it would appear that inhibition of nucleic acid synthesis occurs only at supralethal concentrations of ISCU. The cytotoxicity of ISCU in proliferating or quiescent cell populations was examined. GC3/c1 cells grown in medium containing 0.5% fetal calf serum (FCS) had a 60% reduction in rate of growth, but were more sensitive than cells exposed for 24 h in 10% FCS-medium (IC50 1.9 and 31 micrograms/ml, respectively). When the albumin concentration was adjusted (240 mg/100 ml) to allow equivalent drug binding, IC50 values were similar. In cultures of GC3/c1 cells growth rate was related to the concentration of FCS. In the absence of serum, growth rate was 2.5 to 3.2% that of exponentially growing control cultures in the presence of 10% FCS. Addition of FCS to quiescent cultures after 1 to 6 days in serum-free conditions resulted in immediate growth of cells. Clonogenic potential was also unchanged for at least 6 days under serum-free conditions. Under these conditions, where albumin concentration was adjusted to be equivalent to medium containing 10% FCS, sensitivity of proliferatively quiescent GC3/c1 cells was similar to that in exponentially growing control cultures in which the population doubling time was approximately 22 h. Further, there was no recovery of clonogenic potential when cells were exposed for 24 h to ISCU and maintained in a quiescent state for up to 4 days prior to serum stimulation. These data suggest that the cytotoxic effects of ISCU are independent of the proliferative state of the cell population.(ABSTRACT TRUNCATED AT 400 WORDS)


The role of milk- and soy-based protein in support of muscle protein synthesis and muscle protein accretion in young and elderly persons.
The balance between muscle protein synthesis (MPS) and muscle protein breakdown (MPB) is dependent on protein consumption and the accompanying hyperaminoacidemia, which stimulates a marked rise in MPS and mild suppression of MPB. In the fasting state, however, MPS declines sharply and MPB is increased slightly. Ultimately, the balance between MPS and MPB determines the net rate of muscle growth. Accretion of new muscle mass beyond that of normal growth can occur following periods of intense resistance exercise. Such muscle accretion is an often sought-after goal of athletes. There needs to be, however, an increased appreciation of the role that preservation of muscle can play in offsetting morbidities associated with the sarcopenia of aging, such as type 2 diabetes and declines in metabolic rate that can lead to fat mass accumulation followed by the onset or progression of obesity. Emerging evidence shows that consumption of different types of proteins can have different stimulatory effects on the amplitude and possibly duration that MPS is elevated after feeding; this may be particularly significant after resistance exercise. This effect may be due to differences in the fundamental amino acid composition of the protein (i.e., its amino acid score) and its rate of digestion. Milk proteins, specifically casein and whey, are the highest quality proteins and are quite different in terms of their rates of digestion and absorption. New data suggest that whey protein is better able to support MPS than is soy protein, a finding that may explain the greater ability of whey protein to support greater net muscle mass gains with resistance exercise. This review focuses on evidence showing the differences in responses of MPS, and ultimately muscle protein accretion, to consumption of milk- and soy-based supplemental protein sources in humans.


Synthesis of HIV-1 capsid protein assembly inhibitor (CAP-1) and its analogues based on a biomass approach.
A biomass-derived platform chemical was utilized to access a demanded pharmaceutical substance with anti-HIV activity (HIV, human immunodeficiency virus) and a variety of structural analogues. Step economy in the synthesis of the drug core (single stage from cellulose) is studied including flexible variability of four structural units. The first synthesis and X-ray structure of the inhibitor of HIV-1 capsid protein assembly (CAP-1) is described.


Analgesic abuse, ureteric obstruction, and retroperitoneal fibrosis.
We report two cases of unusual ureteric obstruction in patients with an excessive consumption of analgesics. In a retrospective survey of seven cases of non-malignant retroperitoneal fibrosis seen in the last 15 years it was found that four had taken excessive amounts of analgesics. A careful drug history should be taken in all patients with restroperitoneal fibrosis and ureteric obstruction.


Using Sex Pheromone and a Multi-Scale Approach to Predict the Distribution of a Rare Saproxylic Beetle.
The European red click beetle, Elater ferrugineus L., is associated with wood mould in old hollow deciduous trees. As a result of severe habitat fragmentation caused by human disturbance, it is threatened throughout its distribution range. A new pheromone-based survey method, which is very efficient in detecting the species, was used in the present study to relate the occurrence of E. ferrugineus to the density of deciduous trees. The latter data were from a recently completed regional survey in SE Sweden recording >120,000 deciduous trees. The occurrence of E. ferrugineus increased with increasing amount of large hollow and large non-hollow trees in the surrounding landscape. Quercus robur (oak) was found to be the most important substrate for E. ferrugineus, whereas two groups of tree species (Carpinus betulus, Fagus sylvatica, Ulmus glabra, vs. Acer platanoides, Aesculus hippocastanum, Fraxinus excelsior, Tilia cordata) were less important but may be a complement to oak in sustaining populations of the beetle. The occurrence of E. ferrugineus was explained by the density of oaks at two different spatial scales, within the circle radii 327 m and 4658 m. In conclusion, priority should be given to oaks in conservation management of E. ferrugineus, and then to the deciduous trees in the genera listed above. Conservation planning at large spatial and temporal scales appears to be essential for long-term persistence of E. ferrugineus. We also show that occurrence models based on strategic sampling might result in pessimistic predictions. This study demonstrates how pheromone-based monitoring make insects excellent tools for sustained feedback to models for landscape conservation management.


Elucidating the dual role of grain boundaries as dislocation sources and obstacles and its impact on toughness and brittle-to-ductile transition.
In this paper, we resolve the role of grain boundaries on toughness and the brittle-to-ductile transition. On the one hand, grain boundaries are obstacles for dislocation glide. On the other hand, the intersection points of grain boundaries with the crack front are assumed to be preferred dislocation nucleation sites. Here, we will show that the single contributions of grain boundaries (obstacles vs. source) on toughness and the brittle-to-ductile transition are contradicting, and we will weight the single contributions by performing carefully designed numerical experiments by means of two-dimensional discrete dislocation dynamics modelling. In our parameter studies, we vary the following parameters: (i) the mean free path for dislocation glide, δ, combined with (ii) the (obstacle) force of the grain boundary, ϕ, and (iii) the dislocation source spacing along the crack front, λ. Our results show that for materials or microstructures for which the mean distance of the intersection points of grain boundaries with the crack front is the relevant measure for λ, a decrease of grain size results in an increase of toughness. The positive impact of grain boundaries outweighs the negative consequences of dislocation blocking. Furthermore, our results explain the evolving anisotropy of toughness in cold-worked metals and give further insight into the question of why the grain-size-dependent fracture toughness passes through a minimum (and the brittle-to-ductile transition temperature passes through a maximum) at an intermediate grain size. Finally, a relation of the grain-size-dependence of fracture toughness in the form of K(dδ, dλ) = KIC + kdδ0.5/dλ is deduced.


Plasma fatty acid profile in depressive disorder resembles insulin resistance state.
Depressive disorder is related to an increased risk of type 2 diabetes mellitus (DM2) and cardiovascular disease (CVD). Insulin resistance (IR), connected with altered fatty acid (FA) composition, namely with decreased proportion of polyunsaturated FA could participate in these associations. The aim of the study was to investigate the composition of FA in plasma cholesterol esters (CE) and phosphatidylcholine (PC) as well as indices of insulin resistance and oxidative stress in the patients with depressive disorder. Parameters of lipid and glucose homeostasis, concentrations of FA in plasma cholesteryl esters (CE) and phosphatidylcholine (PC) and conjugated dienes in LDL were investigated in a group of 47 patients (9M/38F) with depression and compared with 47 control persons (16M/31F). Delta-9 desaturase (D9D) and D6D desaturase were estimated as product to precursor fatty acid ratios. In depressive patients increased concentrations of palmitoleic acid and total monounsaturated FA with decreased proportion of total polyunsaturated FA n-6 (PUFA n-6) (all p<0.05) in CE were found, while in PC increased proportion of saturated FA was observed (p<0.05). Moreover, index of D6D activity was significantly increased in PC and CE (p<0.05). Concomitantly, in depressive patients higher levels of plasma triacylglycerols (p<0.05), conjugated dienes in LDL (p<0.001) and HOMA index of IR (p<0.05) were found. Esterified FA composition of depressive patients revealed changes, similar to those, usually observed in insulin resistance. Dysregulation of FA could participate in the pathogenesis of depression and be associated with an increased risk of CVD and DM2.


Ursodeoxycholic acid inhibits TNFα-induced IL-8 release from monocytes.
Monocytes are critical to the pathogenesis of inflammatory bowel disease (IBD) as they infiltrate the mucosa and release cytokines that drive the inflammatory response. Ursodeoxycholic acid (UDCA), a naturally occurring bile acid with anti-inflammatory actions, has been proposed as a potential new therapy for IBD. However, its effects on monocyte function are not yet known. Primary monocytes from healthy volunteers or cultured U937 monocytes were treated with either the proinflammatory cytokine, TNFα (5 ng/ml) or the bacterial endotoxin, lipopolysaccharide (LPS; 1 μg/ml) for 24 h, in the absence or presence of UDCA (25-100 μM). IL-8 release into the supernatant was measured by ELISA. mRNA levels were quantified by qPCR and changes in cell signaling proteins were determined by Western blotting. Toxicity was assessed by measuring lactate dehydrogenase (LDH) release. UDCA treatment significantly attenuated TNFα-, but not LPS-driven, release of IL-8 from both primary and cultured monocytes. UDCA inhibition of TNFα-driven responses was associated with reduced IL-8 mRNA expression. Both TNFα and LPS stimulated NFκB activation in monocytes, while IL-8 release in response to both cytokines was attenuated by an NFκB inhibitor, BMS-345541. Interestingly, UDCA inhibited TNFα-, but not LPS-stimulated, NFκB activation. Finally, TNFα, but not LPS, induced phosphorylation of TNF receptor associated factor (TRAF2), while UDCA cotreatment attenuated this response. We conclude that UDCA specifically inhibits TNFα-induced IL-8 release from monocytes by inhibiting TRAF2 activation. Since such actions would serve to dampen mucosal immune responses in vivo, our data support the therapeutic potential of UDCA for IBD.


[Uncommon cutaneous presentation of visceral Leishmaniasis associated with HIV].
Visceral leishmaniasis is not normally expressed in skin. Herein, we describe the case of an HIV-positive patient who developed two unusual skin manifestations during an episode of visceral leishmaniasis. A 48-year-old female patient consulted initially for infiltrated purpura of all four limbs. Skin biopsy revealed leukocytoclastic vasculitis with Leishman-Donovan bodies. Laboratory tests showed medullary, splenic, gastric and colic involvement, suggesting systemic disease, and enabling visceral leishmaniasis to be diagnosed. Two years later, despite prolonged treatment, the patient presented maculopapular exanthema, and histology revealed persistent Leishman-Donovan bodies. We report herein an association of two rare skin manifestations in an HIV-positive patient with visceral leishmaniasis: infiltrated purpura and maculopapular exanthema. However, the underlying mechanisms remain hypothetical. The initial leukocytoclastic exanthema could be secondary to either polyclonal hypergammaglobulinaemia or to IgA deposits, or possibly to mechanical impairment of blood vessels by the actual parasite. The maculopapular exanthema occurring later raised the possibility of post-Kala-Azar leishmaniasis due to blood-borne dissemination in an anergic subject or perhaps even immune reconstitution inflammatory syndrome.


Towards the Representation of Genomic Data in HL7 FHIR and OMOP CDM.
High throughput sequencing technologies have facilitated an outburst in biological knowledge over the past decades and thus enables improvements in personalized medicine. In order to support (international) medical research with the combination of genomic and clinical patient data, a standardization and harmonization of these data sources is highly desirable. To support this increasing importance of genomic data, we have created semantic mapping from raw genomic data to both FHIR (Fast Healthcare Interoperability Resources) and OMOP (Observational Medical Outcomes Partnership) CDM (Common Data Model) and analyzed the data coverage of both models. For this, we calculated the mapping score for different data categories and the relative data coverage in both FHIR and OMOP CDM. Our results show, that the patients genomic data can be mapped to OMOP CDM directly from VCF (Variant Call Format) file with a coverage of slightly over 50%. However, using FHIR as intermediate representation does not lead to further information loss as the already stored data in FHIR can be further transformed into OMOP CDM format with almost 100% success. Our findings are in favor of extending OMOP CDM with patient genomic data using ETL to enable the researchers to apply different analysis methods including machine learning algorithms on genomic data.


MR neurography: pitfalls in imaging and interpretation.
MR imaging of peripheral nerves, typically referred to as MR neurography, is a rapidly evolving technique that currently is drawing huge attention, both in research and in clinical settings. Both training and experience are necessary to detect the sometimes subtle findings and to avoid misinterpretation of abnormalities. This review article is intended to help radiologists with image evaluation and interpretation. Typical pitfalls are discussed as well as strategies to avoid them.


Characterization of the mechanical properties of cancer cells in 3D matrices in response to collagen concentration and cytoskeletal inhibitors.
Cellular processes, such as cell migration, adhesion, and proliferation depend on the interaction between the intracellular environment and the extracellular matrix (ECM). While many studies have explored the role of the microenvironment on cell behavior, the influence of 3D matrix mechanics on intracellular activity is not completely understood. To characterize the relationship between the mechanical components of the microenvironment and intracellular behavior, we use particle-tracking microrheology of metastatic breast cancer cells embedded in 3D collagen gels to quantify the intracellular activity from which the molecular motor activity and stiffness can be determined. Our results show that increasing collagen concentration of the 3D environments leads to an increase in intracellular stiffness and motor activity. Furthermore, our studies demonstrate that intracellular fluctuations depend on collagen concentration, even in the presence of a number of frontline chemotherapeutic and anti-MMP drugs, indicating that ECM concentration is an important and indispensable parameter to consider in drug screening.


Evolution of immune systems: specificity and autoreactivity.
Multicellularity evolved well before 600 million years ago, and all multicellular animals have evolved since then with the need to protect against pathogens. There is no reason to expect their immune systems to be any less sophisticated than ours. The vertebrate system, based on rearranging immunoglobulin-superfamily domains, appears to have evolved partly as a result of chance insertion of RAG genes by horizontal transfer. Remarkably sophisticated systems for expansion of immunological repertoire have evolved in parallel in many groups of organisms. Vaccination of invertebrates against commercially important pathogens has been empirically successful, and suggests that the definition of an adaptive and innate immune system should no longer depend on the presence of memory and specificity, since these terms are hard to define in themselves. The evolution of randomly-created immunological repertoire also carries with it the potential for generating autoreactive specificities and consequent autoimmune damage. While invertebrates may use systems analogous to ours to control autoreactive specificities, they may have evolved alternative mechanisms which operate either at the level of individuals-within-populations rather than cells-within-individuals, by linking self-reactive specificities to regulatory pathways and non-self-reactive to effector pathways.


Radiofrequency ablation induces antigen-presenting cell infiltration and amplification of weak tumor-induced immunity.
To evaluate the influence of subtotal radiofrequency (RF) ablation on a tumor-specific immune response in a murine tumor model and to explore the role of intratumoral dendritic cells (ITDCs) in mediating this effect. Animal work was performed according to an approved protocol and in compliance with the National Cancer Institute Animal Care and Use Committee guidelines and regulations. A murine urothelial carcinoma (MB49) model expressing the male minor histocompatibility (HY) antigen was inoculated subcutaneously in female mice. Fourteen days later, splenic T cells were analyzed with enzyme-linked immunosorbent spot for HY immune response (n = 57). In subsequent experiments, mice were randomized into control (n = 7), RF ablation, ITDC (n = 9), and RF ablation + ITDC (n = 9) groups and monitored for tumor growth. Eleven days after treatment, tumors were harvested for histologic and immunohistochemical analysis. Animals demonstrating complete tumor regression were rechallenged in the contralateral flank. Animals treated with subtotal RF ablation showed significant increases in tumor-specific class I and II responses to HY antigens and tumor regression. RF ablation, ITDC, and combined groups demonstrated similar levels of antigen-presenting cell infiltration; all groups demonstrated greater levels of infiltration compared with untreated controls. ITDC injection also resulted in tumor regression. However, combination therapy did not enhance tumor regression when compared with either treatment alone. Rechallenged mice in RF ablation, ITDC, and combination groups demonstrated significant tumor growth inhibition compared with controls. Subtotal RF ablation treatment results in enhanced systemic antitumor T-cell immune responses and tumor regression that is associated with increased dendritic cell infiltration. ITDC injection mimics the RF ablation effect but does not increase immune responses when injected immediately after RF ablation.


The role of feeding and nutrition in the pathogeny and prevention of diarrheic processes.
Diarrheic processes pose a grave health threat in much of Latin America, especially for small children. One reason for this is the close connection between diarrhea and lack of proper nutrition. The present article seeks to explore this connection by examining two of its main components: the protection against diarrhea resulting from breast-feeding and the increased vulnerability to diarrhea created by malnutrition. Breast-feeding helps prevent enteric infections in several ways. For one thing, the mother's colostrum and milk contain antibodies against some enterobacterial antigens. For another, the so-called "bifid factor" in human milk helps discourage growth of pathogenic enterobacteria in the intestinal lumen. Furthermore, children living in unhealthy surroundings become heavily exposed to common bacteria when breast-feeding stops, a circumstance deemed largely responsible for "weaning diarrhea." Proper nutrition in general is also important, since diarrhea tends to be more common and severe among malnourished children. Several processes that could contribute to this problem have been suggested. These include morphological alterations of the intestinal mucosa in malnourished children, poor intestinal absorption of fats and other nutrients, irritation caused by increased concentrations of free bile acids, and changes in the composition of the intestinal flora. Though not all these processes are well understood, it is clear that malnutrition favors development of diarrhea, while diarrhea in its turn precipitates and aggravates malnutrition. The sad plight of millions of children in the Americas is the result of this combined interaction. Many of those who experience it die, and the survivors fail to achieve their full potential growth and development. The control of diarrheic infections alone would greatly improve these children's nutritional status. Likewise breast-feeding in the early months of life, duly supplemented later and followed by a sound diet after weaning, would considerably reduce the danger and damage caused by diarrheic infections.


Drinking for protection? Epidemiological and experimental evidence on the beneficial effects of coffee or major coffee compounds against gastrointestinal and liver carcinogenesis.
Recent meta-analyses indicate that coffee consumption reduces the risk for digestive tract (oral, esophageal, gastric and colorectal) and, especially, liver cancer. Coffee bean-derived beverages, as the widely-consumed espresso and "common" filtered brews, present remarkable historical, cultural and economic importance globally. These drinks have rich and variable chemical composition, depending on factors that vary from "seeding to serving". The alkaloids caffeine and trigonelline, as well as the polyphenol chlorogenic acid, are some of the most important bioactive organic compounds of these beverages, displaying high levels in both espresso and common brews and/or increased bioavailability after consumption. Thus, we performed a comprehensive literature overview of current knowledge on the effects of coffee beverages and their highly bioavailable compounds, describing: 1) recent epidemiological and experimental findings highlighting the beneficial effects against gastrointestinal/liver carcinogenesis, and 2) the main molecular mechanisms in these in vitro and in vivo bioassays. Findings predominantly address the protective effects of coffee beverages and their most common/bioavailable compounds individually on gastrointestinal and liver cancer development. Caffeine, trigonelline and chlorogenic acid modulate common molecular targets directly implicated in key cancer hallmarks, what could stimulate novel translational or population-based mechanistic investigations.


Pseudomonas aeruginosa syntrophy in chronically colonized airways of cystic fibrosis patients.
Pseudomonas aeruginosa isolates from cystic fibrosis (CF) patients undergo remarkable phenotypic divergence over time, including loss of pigmentation, hemolysis, motility, and quorum sensing and emergence of antibiotic hypersusceptibility and/or auxotrophism. With prolonged antibiotic treatment and steady decline in lung function in chronically infected patients, the divergent characteristics associated with CF isolates have traditionally been regarded as "adapted/unusual virulence," despite the degenerative nature of these adaptations. We examined the phenotypic and genotypic diversity in clonally related isogenic strains of P. aeruginosa from individual CF patients. Our observations support a novel model of intra-airway pseudomonal syntrophy and accompanying loss of virulence. A 2007 calendar year collection of CF P. aeruginosa isolates (n = 525) from 103 CF patients yielded in vitro MICs of sulfamethoxazole-trimethoprim (SMX-TMP, which typically has no activity against P. aeruginosa) ranging from 0.02 to >32 μg/ml (median, 1.5). Coisolation of clonally related SMX-TMP-susceptible and -resistant P. aeruginosa strains from the same host was common (57%), as were isogenic coisolates with mutations in efflux gene determinants (mexR, mexAB-oprM, and mexZ) and genes governing DNA mismatch repair (mutL and mutS). In this cohort, complete in vitro growth complementation between auxotrophic and prototrophic P. aeruginosa isogenic strains was evident and concurrent with the coding sequence mosaicism in resistance determinants. These observations suggest that syntrophic clonal strains evolve in situ in an organized colonial structure. We propose that P. aeruginosa adopts a multicellular lifestyle in CF patients due to host selection of an energetically favorable, less-virulent microbe restricted within and symbiotic with the airway over the host's lifetime.


Dose-dependent effect of growth hormone on final height in children with short stature without growth hormone deficiency.
The effect of GH therapy in short non-GH-deficient children, especially those with idiopathic short stature (ISS), has not been clearly established owing to the lack of controlled trials continuing until final height (FH). The aim of the study was to investigate the effect on growth to FH of two GH doses given to short children, mainly with ISS, compared with untreated controls. A randomized, controlled, long-term multicenter trial was conducted in Sweden. Two doses of GH (Genotropin) were administered, 33 or 67 microg/kg.d; control subjects were untreated. A total of 177 subjects with short stature were enrolled. Of these, 151 were included in the intent to treat (AllITT) population, and 108 in the per protocol (AllPP) population. Analysis of ISS subjects included 126 children in the ITT (ISSITT) population and 68 subjects in the PP (ISSPP) population. We measured FH sd score (SDS), difference in SDS to midparenteral height (diff MPHSDS), and gain in heightSDS. After 5.9+/-1.1 yr on GH therapy, the FHSDS in the AllPP population treated with GH vs. controls was -1.5+/-0.81 (33 microg/kg.d, -1.7+/-0.70; and 67 microg/kg.d, -1.4+/-0.86; P<0.032), vs. -2.4+/-0.85 (P<0.001); the diff MPHSDS was -0.2+/-1.0 vs. -1.0+/-0.74 (P<0.001); and the gain in heightSDS was 1.3+/-0.78 vs. 0.2+/-0.69 (P<0.001). GH therapy was safe and had no impact on time to onset of puberty. A dose-response relationship identified after 1 yr remained to FH for all growth outcome variables in all four populations. GH treatment significantly increased FH in ISS children in a dose-dependent manner, with a mean gain of 1.3 SDS (8 cm) and a broad range of response from no gain to 3 SDS compared to a mean gain of 0.2 SDS in the untreated controls.


Disordered eating measures validated in pregnancy samples: a systematic review.
Although disordered eating in pregnancy has been linked to numerous negative consequences, there is currently no published instrument specifically devised to identify or measure such symptoms in pregnancy. As such, this study systematically reviewed the literature to evaluate the performance of general measures of disordered eating in pregnancy samples. A systematic search of the following electronic databases was undertaken from inception to April 2019: Scopus, Medline, PsycINFO, Embase, ProQuest Dissertations and Theses, and the Cumulative Index to Nursing and Allied Health Literature. From 1724 citations, eight publications met the inclusion criteria and were included in the review. Most of the included studies (6/8) were of reasonable quality. Overall, three self-report inventories (EDE-Q, EDI-2, and DEBS) and one semi-structured clinical interview (EDE) had some form of psychometric information available. Most studies reported reliability, with only two reporting validity. No studies assessed screening accuracy. Other than the EDE-Q, which had preliminary evidence to suggest possible utility in pregnancy, the findings of this review revealed little to no evidence to support the use of general measures of disordered eating in pregnancy. A strong need for research exploring the validity of existing measures in pregnancy samples, including the EDE-Q, was also evident.


The persistence of Theileria parva infection in cattle immunized using two stocks which differ in their ability to induce a carrier state: analysis using a novel blood spot PCR assay.
An improved Theileria parva DNA detection assay based on the polymerase chain reaction (PCR) using primers derived from the 104 kDa antigen (p104) gene was developed to detect parasite DNA in blood spots on filter paper. The specificity of the assay was validated using DNA from a wide range of cattle-derived and buffalo-derived stocks of T. parva. DNA of T. annulata, T. buffeli, T. lestoquardi, T. mutans and T. taurotragi was not amplified using the p104 primers. The detection threshold of the assay was approximately 1-2 parasites/microl of infected blood. PCR amplification using the p104 primers was applied to sequential samples from groups of cattle experimentally infected with either the T. parva Marikebuni stock that induces a long-term carrier state or the Muguga stock, which does not induce a carrier state. The study extended for up to 487 days post-infection and PCR data from defined time points were compared with parasitological microscopy and serological data, together with xenodiagnosis by experimental application of ticks. Microscopy first detected piroplasms between days 13 and 16 after infection whereas all cattle became PCR +ve between days 9 and 13. Animals infected with the Muguga stock of T. parva had parasite DNA in the peripheral blood, which could be detected by PCR, for between 33 and 129 days post-infection in different animals. By contrast parasite DNA in the blood of cattle infected with the Marikebuni stock could be detected consistently from day 9 up to 487 days, when the study terminated. The data suggest that the nature and persistence of the carrier state may differ markedly between different T. parva parasite stocks.


Structure-Activity Relationships of the MEPicides: N-Acyl and O-Linked Analogs of FR900098 as Inhibitors of Dxr from Mycobacterium tuberculosis and Yersinia pestis.
Despite continued research efforts, the threat of drug resistance from a variety of bacteria continues to plague clinical communities. Discovery and validation of novel biochemical targets will facilitate development of new drugs to combat these organisms. The methylerythritol phosphate (MEP) pathway to make isoprene units is a biosynthetic pathway essential to many bacteria. We and others have explored inhibitors of the MEP pathway as novel antibacterial agents. Mycobacterium tuberculosis, the causative agent of tuberculosis, and Yersinia pestis, resulting in the plague or "black death", both rely on the MEP pathway for isoprene production. 1-Deoxy-d-xylulose 5-phosphate reductoisomerase (Dxr) catalyzes the first committed step in the MEP pathway. We examined two series of Dxr inhibitors based on the parent structure of the retrohydroxamate natural product FR900098. The compounds contain either an extended N-acyl or O-linked alkyl/aryl group and are designed to act as bisubstrate inhibitors of the enzyme. While nearly all of the compounds inhibited both Mtb and Yp Dxr to some extent, compounds generally displayed more potent inhibition against the Yp homologue, with the best analogs displaying nanomolar IC50 values. In bacterial growth inhibition assays, the phosphonic acids generally resulted in poor antibacterial activity, likely a reflection of inadequate permeability. Accordingly, diethyl and dipivaloyloxymethyl (POM) prodrug esters of these compounds were made. While the added lipophilicity did not enhance Yersinia activity, the compounds showed significantly improved antitubercular activities. The most potent compounds have Mtb MIC values of 3-12 μg/mL. Taken together, we have uncovered two series of analogs that potently inhibit Dxr homologues from Mtb and Yp. These inhibitors of the MEP pathway, termed MEPicides, serve as leads for future analog development.


Recent partner violence and sexual and drug-related STI/HIV risk among adolescent and young adult women attending family planning clinics.
Adolescent and young adult women are at high risk for sexually transmitted infections (STIs)/HIV and intimate partner violence (IPV). We evaluate the prevalence of IPV in the past 3 months and its associations with STI/HIV risk, STI and related care-seeking over the same time period. Female family planning clinic patients ages 16-29 years (n=3504) participated in a cross-sectional survey in 2011-2012 as a baseline assessment for an intervention study. We examined associations of recent IPV with sexual and drug-related STI/HIV risk behaviour, self-reported STI and STI-related clinical care seeking via logistic regression. Recent physical or sexual IPV (prevalence 11%) was associated with recent sexual and drug-related STI/HIV risk, specifically unprotected vaginal sex (adjusted OR (AOR) 1.93, 95% CI 1.52 to 2.44), unprotected anal sex (AOR 2.22, 95% CI 1.51 to 3.27) and injection drug use, their own (AOR 3.39, 95% CI 1.47 to 7.79) and their partner's (AOR 3.85, 1.91 to 7.75). IPV was also linked with coercive sexual risk: involuntary condom non-use (AOR 1.87 to 95% CI 1.51 to 2.33), and fears of requesting condoms (AOR 4.15, 95% CI 2.73 to 6.30) and refusing sex (AOR 11.84, 95% CI 7.59 to 18.45). STI-related care-seeking was also more common among those abused (AOR 2.49, 95% CI 1.87 to 3.31). Recent IPV is concurrent with sexual and drug-related STI/HIV risk, including coercive sexual risk, thus compromising women's agency in STI/HIV risk reduction. Clinical risk assessments should broaden to include unprotected heterosexual anal sex, coercive sexual risk and IPV, and should promote safety and harm reduction.


Multi-Omics Studies Demonstrate Toxoplasma gondii-Induced Metabolic Reprogramming of Murine Dendritic Cells.
Toxoplasma gondii is capable of actively invading almost any mammalian cell type including phagocytes. Early events in phagocytic cells such as dendritic cells are not only key to establishing parasite infection, but conversely play a pivotal role in initiating host immunity. It is now recognized that in addition to changes in canonical immune markers and mediators, alteration in metabolism occurs upon activation of phagocytic cells. These metabolic changes are important for supporting the developing immune response, but can affect the availability of nutrients for intracellular pathogens including T. gondii. However, the interaction of T. gondii with these cells and particularly how infection changes their metabolism has not been extensively investigated. Herein, we use a multi-omics approach comprising transcriptomics and metabolomics validated with functional assays to better understand early events in these cells following infection. Analysis of the transcriptome of T. gondii infected bone marrow derived dendritic cells (BMDCs) revealed significant alterations in transcripts associated with cellular metabolism, activation of T cells, inflammation mediated chemokine and cytokine signaling pathways. Multivariant analysis of metabolomic data sets acquired through non-targeted liquid chromatography mass spectroscopy (LCMS) identified metabolites associated with glycolysis, the TCA cycle, oxidative phosphorylation and arginine metabolism as major discriminants between control uninfected and T. gondii infected cells. Consistent with these observations, glucose uptake and lactate dehydrogenase activity were upregulated in T. gondii infected BMDC cultures compared with control BMDCs. Conversely, BMDC mitochondrial membrane potential was reduced in T. gondii-infected cells relative to mitochondria of control BMDCs. These changes to energy metabolism, similar to what has been described following LPS stimulation of BMDCs and macrophages are often termed the Warburg effect. This metabolic reprogramming of cells has been suggested to be an important adaption that provides energy and precursors to facilitate phagocytosis, antigen processing and cytokine production. Other changes to BMDC metabolism are evident following T. gondii infection and include upregulation of arginine degradation concomitant with increased arginase-1 activity and ornithine and proline production. As T. gondii is an arginine auxotroph the resultant reduced cellular arginine levels are likely to curtail parasite multiplication. These results highlight the complex interplay of BMDCs and parasite metabolism within the developing immune response and the consequences for adaptive immunity and pathogen clearance.


Approved and investigational uses of modafinil : an evidence-based review.
Modafinil is a wake-promoting agent that is pharmacologically different from other stimulants. It has been investigated in healthy volunteers, and in individuals with clinical disorders associated with excessive sleepiness, fatigue, impaired cognition and other symptoms. This review examines the use of modafinil in clinical practice based on the results of randomized, double-blind, placebo-controlled clinical trials available in the English language in the MEDLINE database. In sleep-deprived individuals, modafinil improves mood, fatigue, sleepiness and cognition to a similar extent as caffeine but has a longer duration of action. Evidence for improved cognition in non-sleep-deprived healthy volunteers is controversial.Modafinil improves excessive sleepiness and illness severity in all three disorders for which it has been approved by the US FDA, i.e. narcolepsy, shift-work sleep disorder and obstructive sleep apnoea with residual excessive sleepiness despite optimal use of continuous positive airway pressure (CPAP). However, its effects on safety on the job and on morbidities associated with these disorders have not been ascertained. Continued use of CPAP in obstructive sleep apnoea is essential. Modafinil does not benefit cataplexy.In very small, short-term trials, modafinil improved excessive sleepiness in patients with myotonic dystrophy. It was efficacious in fairly large studies of attention deficit hyperactivity disorder (ADHD) in children and adolescents, and was as efficacious as methylphenidate in a small trial, but has not been approved by the FDA, in part because of its serious dermatological toxicity. In a trial of 21 non-concurrent subjects, with 2-week treatment periods, modafinil was as effective as dexamfetamine in adult ADHD. Modafinil was helpful for depressive symptoms in bipolar disorder in a trial that excluded patients with stimulant-induced mania. A single dose of modafinil may hasten recovery from general anaesthesia after day surgery. A single dose of modafinil improved the ability of emergency room physicians to attend didactic lectures after a night shift, but did not improve their ability to drive home and caused sleep disturbances subsequently.Modafinil had a substantial placebo effect on outcomes such as fatigue, excessive sleepiness and depression in patients with traumatic brain injury, major depressive disorder, schizophrenia, post-polio fatigue and multiple sclerosis; however, it did not provide any benefit greater than placebo.Trials of modafinil for excessive sleepiness in Parkinson's disease, cocaine addiction and cognition in chronic fatigue syndrome provided inconsistent results; all studies had extremely small sample sizes. Modafinil cannot be recommended for these conditions until definitive data become available.Modafinil induces and inhibits several cytochrome P450 isoenzymes and has the potential for interacting with drugs from all classes. The modafinil dose should be reduced in the elderly and in patients with hepatic disease. Caution is needed in patients with severe renal insufficiency because of substantial increases in levels of modafinil acid. Common adverse events with modafinil include insomnia, headache, nausea, nervousness and hypertension. Decreased appetite, weight loss and serious dermatological have been reported with greater frequency in children and adolescents, probably due to the higher doses (based on bodyweight) used. Modafinil may have some abuse/addictive potential although no cases have been reported to date.


B type natriuretic peptide--a diagnostic breakthrough in peri-operative cardiac risk assessment?
The B-type natriuretic peptides; B-type natriuretic peptide and N-terminal pro-B-type natriuretic peptide, are increasing being used as biomarkers for the diagnosis, management and prognostication of cardiac failure, but their application in the peri-operative period is unclear. This review examines the current understanding of the role of B-type natriuretic peptides in both the operative and non-operative settings. Normal values, diagnostic thresholds, monitoring targets and significant prognostic levels are identified. Using this as a background, the role of B-type natriuretic peptides in the prediction of peri-operative mortality and morbidity is examined and potential confounders, such as renal failure and body mass index, which may impact significantly on the utility of the biomarkers, are discussed. Clinical recommendations with regard to its use are made and a research agenda is proposed for future peri-operative studies.


The Most Effective Methods for Delivering Severe Weather Early Warnings to Fishermen on Lake Victoria.
Introduction: It is estimated that five thousand people die on Lake Victoria every year by drowning which is triggered by severe weather hazards like lightning. Objectives: In order to improve predictability of severe weather conditions on Lake Victoria, there is need to deliver timely and effective Severe Weather Early Warning Systems (SWEWS) to those at risk. On Lake Victoria, previous SWEW service trials ceased with the end of the funding grants. This study therefore assessed the possibility of sustaining the SWEW service by assessing willingness to pay. Methods: An assessment was conducted between March and May 2015 to determine the SWEW service improvements desired by the population. A convenience sample of respondents was gathered and interviewed during impromptu visits to landing sites on Lake Victoria. The respondents were also among community members that had earlier participated in a pilot assessing the feasibility of mobile phones is delivering SWEW alerts. Semi-structured questionnaires were administered to fishermen and fisher folks at the landing site to gather suggestions/strategies for (i) better design and implementation of SWEW service, (ii) use of smart phones, and (iii) their ability and willingness to pay for a SWEW service. Results were presented as frequencies. Results: Two hundred fifteen respondents from fourteen landing sites (communities) were interviewed. Over 50% of the respondents (113/215) were aware about at least one community member who had been injured due to lightening on the lake in the past year. Ninety two percent (198/215) of the respondents reported using mobile phones as their main tool of communication but only 4% had smart phones that could receive early warning weather alerts through internet connectivity. Seventy five percent of respondents said they would welcome a system that could deliver commercial weather alerts and 65% were willing to pay for such a service. Conclusions: A SWEW service is feasible in this community but must be accompanied with public education on risk, a design that can fit the basic phone functionality and a system that the community majority will be willing to pay for on a continuing basis as a sustainability plan/strategy for an early warning system. This will enable timely dissemination of severe weather alerts and reduce risk of drowning on lakes among fishing communities.


Effects of low-intensity pulsed ultrasound on osteoclasts: Analysis with goldfish scales as a model of bone.
The effects of low-intensity pulsed ultrasound (LIPUS) on osteoclastogenesis were examined using fish scales that had both osteoclasts and osteoblasts. The binding of the receptor activator of NF-κB ligand (RANKL) in osteoblasts to the receptor activator of NF-κB (RANK) in osteoclasts induced osteoclastogenesis. Therefore, we focused on RANK/RANKL signaling. After 6 h of incubation following LIPUS treatment, mRNA expression of RANKL increased significantly. Resulting from the increased RANKL mRNA level, the expression of transcription-regulating factors significantly increased after 6 h of incubation, and then the mRNA expression of functional genes was significantly up-regulated after 12 h of incubation. However, the mRNA expression of osteoprotegerin (OPG), which is known as an osteoclastogenesis inhibitory factor, also significantly increased after 6 h of incubation and tended to further increase after 12 h of incubation. At 24 h of incubation, osteoclastic functional genes' mRNA expression decreased to the level of the control. Furthermore, we performed an in vivo experiment with goldfish. Two weeks after daily LIPUS exposure, osteoclastic marker enzymes tended to decrease while osteoblastic marker enzymes were activated. The regeneration rate of the LIPUS-treated scales was significantly higher than that of the control scales. Thus, LIPUS moderately activates osteoclasts and induces bone formation.


The TSC1-TSC2 complex is required for proper activation of mTOR complex 2.
The mammalian target of rapamycin (mTOR) is a protein kinase that forms two functionally distinct complexes important for nutrient and growth factor signaling. Both complexes phosphorylate a hydrophobic motif on downstream protein kinases, which contributes to the activation of these kinases. mTOR complex 1 (mTORC1) phosphorylates S6K1, while mTORC2 phosphorylates Akt. The TSC1-TSC2 complex is a critical negative regulator of mTORC1. However, how mTORC2 is regulated and whether the TSC1-TSC2 complex is involved are unknown. We find that mTORC2 isolated from a variety of cells lacking a functional TSC1-TSC2 complex is impaired in its kinase activity toward Akt. Importantly, the defect in mTORC2 activity in these cells can be separated from effects on mTORC1 signaling and known feedback mechanisms affecting insulin receptor substrate-1 and phosphatidylinositol 3-kinase. Our data also suggest that the TSC1-TSC2 complex positively regulates mTORC2 in a manner independent of its GTPase-activating protein activity toward Rheb. Finally, we find that the TSC1-TSC2 complex can physically associate with mTORC2 but not mTORC1. These data demonstrate that the TSC1-TSC2 complex inhibits mTORC1 and activates mTORC2, which through different mechanisms promotes Akt activation.


Direct therapeutic applications of calcium electroporation to effectively induce tumor necrosis.
Electroporation of cells with short, high-voltage pulses causes a transient permeabilization of cell membranes that permits passage of otherwise nonpermeating ions and molecules. In this study, we illustrate how electroporation with isotonic calcium can achieve highly effective cancer cell kill in vivo. Calcium electroporation elicited dramatic antitumor responses in which 89% of treated tumors were eliminated. Histologic analyses indicated complete tumor necrosis. Mechanistically, calcium electroporation caused acute ATP depletion likely due to a combination of increased cellular use of ATP, decreased production of ATP due to effects on the mitochondria, as well as loss of ATP through the permeabilized cell membrane. Taken together, our findings offer a preclinical proof of concept for the use of electroporation to load cancer cells with calcium as an efficient anticancer treatment. Electroporation equipment is already used clinically to enhance the delivery of chemotherapy to superficial tumors, with trials on internal tumors in progress, enabling the introduction of calcium electroporation to clinical use. Moreover, the safety profile, availability, and low cost of calcium facilitate access to this technology for many cancer patients in developed and developing countries.


Scientific reporting is suboptimal for aspects that characterize genetic risk prediction studies: a review of published articles based on the Genetic RIsk Prediction Studies statement.
Our main objective was to raise awareness of the areas that need improvements in the reporting of genetic risk prediction articles for future publications, based on the Genetic RIsk Prediction Studies (GRIPS) statement. We evaluated studies that developed or validated a prediction model based on multiple DNA variants, using empirical data, and were published in 2010. A data extraction form based on the 25 items of the GRIPS statement was created and piloted. Forty-two studies met our inclusion criteria. Overall, more than half of the evaluated items (34 of 62) were reported in at least 85% of included articles. Seventy-seven percentage of the articles were identified as genetic risk prediction studies through title assessment, but only 31% used the keywords recommended by GRIPS in the title or abstract. Seventy-four percentage mentioned which allele was the risk variant. Overall, only 10% of the articles reported all essential items needed to perform external validation of the risk model. Completeness of reporting in genetic risk prediction studies is adequate for general elements of study design but is suboptimal for several aspects that characterize genetic risk prediction studies such as description of the model construction. Improvements in the transparency of reporting of these aspects would facilitate the identification, replication, and application of genetic risk prediction models.


Neutrophil to lymphocyte ratio is associated with more extensive, severe and complex coronary artery disease and impaired myocardial perfusion.
We investigated the relation between neutrophil to lymphocyte ratio (N/L) and the extent, severity, and complexity of coronary artery disease (CAD) and myocardial perfusion. One hundred and fifty-one patients who underwent coronary angiography with stable angina pectoris (SAP) (n=93) or acute coronary syndrome (ACS) (n=58) were included in the study. Blood samples were drawn before coronary angiography. Gensini and SYNTAX scores and myocardial blush grade (MBG) were assessed. Neutrophil counts were 4.4±1.4 and 5.0±1.6 in the SAP and ACS groups (p=0.018), whereas lymphocyte counts were 2.2±0.7 and 2.1±0.7, respectively (p=0.104). N/L was 2.2±1.2 in the SAP and 2.6±1.0 in the ACS (p=0.002) groups. In patients with SAP, N/L was significantly correlated with Gensini and SYNTAX scores (Gensini score r=0.32, p=0.002; SYNTAX score r=0.36, p=0.000), but there was no significant correlation between N/L and MBG. In the ACS group, N/L had a more powerful association with both Gensini and SYNTAX scores (Gensini r=0.42, p=0.001; SYNTAX r=0.51, p=0.000). N/L was negatively correlated with MBG in ACS patients (r= -0.48, p=0.000). Significant correlations persisted both in the SAP and ACS groups after correcting for age, diabetes, hyperlipidemia, and statin use; however, the associations were weaker. Cut-off N/L to predict moderate to severe CAD according to SYNTAX score was 2.26, with 72% sensitivity and 71% specificity (area under the curve [AUC]: 0.772, 95% confidence interval [CI] 0.679-0.865, p<0.001). N/L is associated with severe, extensive and complex CAD and may be used to predict moderate to severe involvement in patients with CAD.


Self-assembling Fmoc dipeptide hydrogel for in situ 3D cell culturing.
Conventional cell culture studies have been performed on 2D surfaces, resulting in flat, extended cell growth. More relevant studies are desired to better mimic 3D in vivo tissue growth. Such realistic environments should be the aim of any cell growth study, requiring new methods for culturing cells in vitro. Cell biology is also tending toward miniaturization for increased efficiency and specificity. This paper discusses the application of a self-assembling peptide-derived hydrogel for use as a 3D cell culture scaffold at the microscale. Phenylalanine derivative hydrogel formation was seen to occur in multiple dispersion media. Cells were immobilized in situ within microchambers designed for cell analysis. Use of the highly biocompatible hydrogel components and simplistic procedures significantly reduced the cytotoxic effects seen with alternate 3D culture materials and microstructure loading methods. Cells were easily immobilized, sustained and removed from microchambers. Differences in growth morphology were seen in the cultured cells, owing to the 3-dimentional character of the gel structure. Degradation improved the removal of hydrogel from the microstructures, permitting reuse of the analysis platforms. Self-assembling diphenylalanine derivative hydrogel provided a method to dramatically reduce the typical difficulties of microculture formation. Effective generation of patterned 3D cultures will lead to improved cell study results by better modeling in vivo growth environments and increasing efficiency and specificity of cell studies. Use of simplified growth scaffolds such as peptide-derived hydrogel should be seen as highly advantageous and will likely become more commonplace in cell culture methodology.


Presbiphonya.
This article attempts to describe the aging process of the vocal folds and the main features of the aged voice. In the world ageing population era, aging diseases and aging disorders are crucial. Voice disorders (presbyphonia) are common in the elderly and have a significant impact on communication and quality of life. Some of these disorders depend on the vocal folds, which consist of an extracellular matrix (ECM), fibrous proteins, interstitial proteins, and glycosaminoglycans. The density and spatial arrangement of these elements are important, as changes in their deposition can alter the biomechanical properties and vibratory function of the vocal folds. The aging voice process is analyzed in detail from mechanical factors like pulmonary bellows alteration, to hormonal factors and life style. The elderly people undergoe mechanical, anatomical and functional changes: alterations of the pulmonary bellows, systemic changes like hormonal disregulation, and laryngeal changes, that resulting in hoarseness, which is difficult to treat.


The Bacterial Effector AvrPto Targets the Regulatory Coreceptor SOBIR1 and Suppresses Defense Signaling Mediated by the Receptor-Like Protein Cf-4.
Receptor-like proteins (RLPs) and receptor-like kinases (RLKs) are cell-surface receptors that are essential for detecting invading pathogens and subsequent activation of plant defense responses. RLPs lack a cytoplasmic kinase domain to trigger downstream signaling leading to host resistance. The RLK SOBIR1 constitutively interacts with the tomato RLP Cf-4, thereby providing Cf-4 with a kinase domain. SOBIR1 is required for Cf-4-mediated resistance to strains of the fungal tomato pathogen Cladosporium fulvum that secrete the effector Avr4. Upon perception of this effector by the Cf-4/SOBIR1 complex, the central regulatory RLK SOMATIC EMBRYOGENESIS RECEPTOR KINASE 3a (SERK3a) is recruited to the complex and defense signaling is triggered. SOBIR1 is also required for RLP-mediated resistance to bacterial, fungal ,and oomycete pathogens, and we hypothesized that SOBIR1 is targeted by effectors of such pathogens to suppress host defense responses. In this study, we show that Pseudomonas syringae pv. tomato DC3000 effector AvrPto interacts with Arabidopsis SOBIR1 and its orthologs of tomato and Nicotiana benthamiana, independent of SOBIR1 kinase activity. Interestingly, AvrPto suppresses Arabidopsis SOBIR1-induced cell death in N. benthamiana. Furthermore, AvrPto compromises Avr4-triggered cell death in Cf-4-transgenic N. benthamiana, without affecting Cf-4/SOBIR1/SERK3a complex formation. Our study shows that the RLP coreceptor SOBIR1 is targeted by a bacterial effector, which results in compromised defense responses.


[Phadiatop in the diagnosis of respiratory allergy in children: Allergy Project--PROAL].
To evaluate the positivity of Phadiatop in children from several Brazilian pediatric allergology centers and to compare its results with the presence of serum specific IgE to inhalant and food allergens. Phadiatop and serum specific IgE levels (RAST) to inhalant and food allergens (UniCAP - Pharmacia) were measured in 457 children from several pediatric allergy centers and in a non-allergic control group (n = 62), distributed across five age groups. Phadiatop was positive in 305 atopic children (67.6%) and in 25.8% of controls (p < 0.001). Among atopic children the distribution of positive test varied according to age: 7.9% (24/305) among under 2 year-olds, 15.4% (47/305) in 2 to 3 year-olds, 22.0% (67/305) in 3 to 4 year-olds, 19.3% (59/305) in 4 to 5 year-olds and 35.4% (108/305) in 5 to 12 year-olds. No concordance between food allergens and Phadiatop was observed. Analysis of the relationship between positive inhaled allergen RASTs and positive Phadiatop showed best indices with house dust mites (D. pteronyssinus, D. farinae and Blomia tropicalis). Phadiatop is a useful tool for diagnosing domestic mite allergy.


Crystal structure of the catalytic domain of the PknB serine/threonine kinase from Mycobacterium tuberculosis.
With the advent of the sequencing programs of prokaryotic genomes, many examples of the presence of serine/threonine protein kinases in these organisms have been identified. Moreover, these kinases could be classified as homologues of those belonging to the well characterized superfamily of the eukaryotic serine/threonine and tyrosine kinases. Eleven such kinases were recognized in the genome of Mycobacterium tuberculosis. Here we report the crystal structure of an active form of PknB, one of the four M. tuberculosis kinases that are conserved in the downsized genome of Mycobacterium leprae and are therefore presumed to play an important role in the processes that regulate the complex life cycle of mycobacteria. Our structure confirms again the extraordinary conservation of the protein kinase fold and constitutes a landmark that extends this conservation across the evolutionary distance between high eukaryotes and eubacteria. The structure of PknB, in complex with a nucleotide triphosphate analog, reveals an enzyme in the active state with an unprecedented arrangement of the Gly-rich loop associated with a new conformation of the nucleotide gamma-phosphoryl group. It presents as well a partially disordered activation loop, suggesting an induced fit mode of binding for the so far unknown substrates of this kinase or for some modulating factor(s).


Acute damage to the posterior limb of the internal capsule on diffusion tensor tractography as an early imaging predictor of motor outcome after stroke.
Early prediction of motor outcome is of interest in stroke management. We aimed to determine whether lesion location at DTT is predictive of motor outcome after acute stroke and whether this information improves the predictive accuracy of the clinical scores. We evaluated 60 consecutive patients within 12 hours of middle cerebral artery stroke onset. We used DTT to evaluate CST involvement in the motor cortex and premotor cortex, centrum semiovale, corona radiata, and PLIC and in combinations of these regions at admission, at day 3, and at day 30. Severity of limb weakness was assessed by using the motor subindex scores of the National Institutes of Health Stroke Scale (5a, 5b, 6a, 6b). We calculated volumes of infarct and fractional anisotropy values in the CST of the pons. Acute damage to the PLIC was the best predictor associated with poor motor outcome, axonal damage, and clinical severity at admission (P < .001). There was no significant correlation between acute infarct volume and motor outcome at day 90 (P = .176, r = 0.485). The sensitivity, specificity, and positive and negative predictive values of acute CST involvement at the level of the PLIC for motor outcome at day 90 were 73.7%, 100%, 100%, and 89.1%, respectively. In the acute stage, DTT predicted motor outcome at day 90 better than the clinical scores (R(2) = 75.50, F = 80.09, P < .001). In the acute setting, DTT is promising for stroke mapping to predict motor outcome. Acute CST damage at the level of the PLIC is a significant predictor of unfavorable motor outcome.


The Reliability and Validity of a Four-Minute Running Time-Trial in Assessing [Formula: see text]max and Performance.
Introduction: Traditional graded-exercise tests to volitional exhaustion (GXTs) are limited by the need to establish starting workloads, stage durations, and step increments. Short-duration time-trials (TTs) may be easier to implement and more ecologically valid in terms of real-world athletic events. The purpose of the current study was to assess the reliability and validity of maximal oxygen uptake ([Formula: see text]max) and performance measured during a traditional GXT (STEP) and a four-minute running time-trial (RunTT). Methods: Ten recreational runners (age: 32 ± 7 years; body mass: 69 ± 10 kg) completed five STEP tests with a verification phase (VER) and five self-paced RunTTs on a treadmill. The order of the STEP/VER and RunTT trials was alternated and counter-balanced. Performance was measured as time to exhaustion (TTE) for STEP and VER and distance covered for RunTT. Results: The coefficient of variation (CV) for [Formula: see text]max was similar between STEP, VER, and RunTT (1.9 ± 1.0, 2.2 ± 1.1, and 1.8 ± 0.8%, respectively), but varied for performance between the three types of test (4.5 ± 1.9, 9.7 ± 3.5, and 1.8 ± 0.7% for STEP, VER, and RunTT, respectively). Bland-Altman limits of agreement (bias ± 95%) showed [Formula: see text]max to be 1.6 ± 3.6 mL·kg-1·min-1 higher for STEP vs. RunTT. Peak HR was also significantly higher during STEP compared with RunTT (P = 0.019). Conclusion: A four-minute running time-trial appears to provide more reliable performance data in comparison to an incremental test to exhaustion, but may underestimate [Formula: see text]max.


Intracellular inactivation of thyroid hormone is a survival mechanism for muscle stem cell proliferation and lineage progression.
Precise control of the thyroid hormone (T3)-dependent transcriptional program is required by multiple cell systems, including muscle stem cells. Deciphering how this is achieved and how the T3 signal is controlled in stem cell niches is essentially unknown. We report that in response to proliferative stimuli such as acute skeletal muscle injury, type 3 deiodinase (D3), the thyroid hormone-inactivating enzyme, is induced in satellite cells where it reduces intracellular thyroid signaling. Satellite cell-specific genetic ablation of dio3 severely impairs skeletal muscle regeneration. This impairment is due to massive satellite cell apoptosis caused by exposure of activated satellite cells to the circulating TH. The execution of this proapoptotic program requires an intact FoxO3/MyoD axis, both genes positively regulated by intracellular TH. Thus, D3 is dynamically exploited in vivo to chronically attenuate TH signaling under basal conditions while also being available to acutely increase gene programs required for satellite cell lineage progression.


Pharmacological activation of kainate receptors drives endocannabinoid mobilization.
Activation of both presynaptic metabotropic cannabinoid type 1 receptors (CB(1)s) and ionotropic kainate receptors (KARs) can efficiently modulate GABA release at many synapses of the CNS. The inhibitory effect of kainic acid (KA) has been ascribed to metabotropic actions, and KAR-induced release of secondary neuromodulatory agents may partly mediate these actions. Here, we investigated the involvement of the endocannabinoid system in the modulation of GABAergic synaptic transmission by pharmacological activation of KARs with KA in CA1 pyramidal neurons of the mouse hippocampus. We show that the depression of GABAergic synaptic transmission induced by KA (3 μm) is strongly inhibited by the simultaneous blockade of CB(1) and GABA(B) receptors with SR141716A (5 μm) and CGP55845 (5 μm), respectively. KA induces a calcium-dependent mobilization of the endocannabinoid anandamide (AEA) by activation of GluK2-containing KARs in postsynaptic pyramidal neurons. Consistently, the effect of KA is prolonged by the inhibitor of AEA degradation URB597 (1 μm) in a CB(1)-dependent manner, but it is not altered by blockade of degradation or synthesis of the other main endocannabinoid 2-arachidonoylglycerol (2AG). Hence, our work reveals that the pharmacological activation of KARs leads to the stimulation of secondary metabotropic signaling systems. In addition, these data further underline the profound mechanistic differences between exogenous and endogenous activation of KARs in the hippocampus.


Safety of Repeated Open-Label Treatment Courses of Intravenous Ofatumumab, a Human Anti-CD20 Monoclonal Antibody, in Rheumatoid Arthritis: Results from Three Clinical Trials.
To investigate the safety of ofatumumab retreatment in rheumatoid arthritis. Patients with active rheumatoid arthritis participating in two phase III trials (OFA110635 and OFA110634) and a phase II extension trial (OFA111752) received individualised open-label ofatumumab retreatment (700 mg X 2 intravenous infusions two weeks apart) ≥24 weeks following the first course and ≥16 weeks following further courses. Retreatment required evidence of clinical response followed by disease relapse. These studies were prematurely terminated by the sponsor to refocus development on subcutaneous delivery. Due to differences in study designs and populations, data are summarised separately for each study. 483 patients (243, 148 and 92 in OFA110635, OFA110634 and OFA111752 respectively) received up to 7 treatment courses of intravenous ofatumumab; cumulative duration of exposure was 463, 182 and 175 patient-years, respectively. Mean time between courses was 17-47 weeks. Ofatumumab induced a profound depletion of peripheral B-lymphocytes. Retreated patients derived benefit based on improvement in DAS28. Adverse events were reported for 93% (226/243), 91% (134/148) and 76% (70/92), serious adverse events for 18% (44/243), 20% (30/148) and 12% (11/92) and serious infections for 3% (8/243), 5% (7/148) and 1% (1/92) of patients in OFA110635, OFA110634 and OFA111752, respectively. The most common adverse events were infusion-related reactions during the first infusion of the first course (48-79%); serious infusion-related reactions were rare (<1% [1/243], 5% [8/148], and 1% [1/92] of patients). Two deaths occurred (fulminant hepatitis B virus infection and interstitial lung disease). Ofatumumab was generally well tolerated with no evidence of increased safety risks with multiple retreatments. Serious infections were uncommon and did not increase over time. ClinicalTrials.gov 110635 ClinicalTrials.gov 110634 ClinicalTrials.gov 111752.


Region-specific developmental specialization of GABA-glycine cosynapses in laminas I-II of the rat spinal dorsal horn.
The spinal dorsal horn is the first level of the CNS in which nociceptive input from sensory afferents is integrated and transmitted. Although inhibitory control in this region has a crucial impact on pain transmission, the respective contribution of GABA and glycine to this inhibition remains elusive. We have previously documented co-release of GABA and glycine at the same inhibitory synapse in spinal laminas I-II of adult rats [older than postnatal day 30 (P30)]. However, despite this co-release, individual miniature inhibitory postsynaptic currents (mIPSCs) were mediated by either glycine receptors (GlyR) or GABA(A) receptors (GABA(A)R), yet never by the two together. In contrast, recent studies of ventral horn immature inhibitory synapses (</=P21) reported individual mIPSCs that were mediated by both GABA(A)Rs and GlyRs. This raises the question of whether mixed mIPSCs are present in immature lamina I-II neurons yet are lost through a maturation-dependent synaptic specialization. To test this, we recorded mIPSCs using patch-clamp techniques in lamina I-II neurons in spinal slices taken at different stages of development. We found that, in neurons younger than P23, both GlyR-only and GABA(A)R-only mIPSCs could be recorded, in addition to mixed GABA(A)R-GlyR mIPSCs. With maturation however, both lamina I-II neurons gradually discontinued exhibiting mixed mIPSCs, although with differing patterns of specialization. Yet, at all developmental stages, benzodiazepine administration could unmask mixed mIPSCs. Together, these findings indicate that, although GABA and glycine are continually co-released throughout development, junctional codetection ceases by adulthood. This indicates an age-dependent postsynaptic tuning of inhibitory synapses that occurs in a region-specific manner.


Sequence typing and comparison of population biology of Campylobacter coli and Campylobacter jejuni.
A multilocus sequence typing (MLST) scheme that uses the same loci as a previously described system for Campylobacter jejuni was developed for Campylobacter coli. The C. coli-specific primers were validated with 53 isolates from humans, chickens, and pigs, together with 15 Penner serotype reference isolates. The nucleotide sequence of the flaA short variable region (SVR) was determined for each isolate. These sequence data were compared to equivalent information for 17 C. jejuni isolates representing the known genetic diversity of this species. C. coli and C. jejuni share approximately 86.5% identity at the nucleotide sequence level within the MLST loci. There is evidence of genetic exchange of the housekeeping genes between the two species, but at a very low rate; only one sequence type from each species showed evidence of imported DNA. The flaA gene was more variable and has been exchanged many times between the two species, making it an unreliable marker for species identification but useful for distinguishing closely related strains. All but 3 of 21 human C. coli clinical isolates were distinct, according to the combined MLST and SVR sequences. The use of a common MLST scheme allows direct comparisons of the population biology and molecular epidemiology of these two closely related human pathogens.


Prostatic urethral lift for the treatment of lower urinary tract symptoms in men with benign prostatic hyperplasia.
A variety of minimally invasive surgical approaches are available as an alternative to transurethral resection of prostate (TURP) for the management of lower urinary tract symptoms (LUTS) in men with benign prostatic hyperplasia (BPH). A recent addition to these is prostatic urethral lift (PUL). To assess the effects of PUL for the treatment of LUTS in men with BPH. We performed a comprehensive search of multiple databases (the Cochrane Library, MEDLINE, Embase, LILACS, Scopus, Web of Science, and Google Scholar), trials registries, other sources of grey literature, and conference proceedings with no restrictions on the language of publication or publication status up until 31 January 2019. We included parallel group randomized controlled trials (RCTs). While we planned to include non-RCTs if RCTs had provided low-certainty evidence for a given outcome and comparison, we could not find any non-RCTs. Two review authors independently screened the literature, extracted data, and assessed risk of bias. We performed statistical analyses using a random-effects model and interpreted them according to the Cochrane Handbook for Systematic Reviews of Interventions. We planned subgroup analyses by age, prostate volume, and severity of baseline symptoms. We used the GRADE approach to rate the certainty of the evidence. We included two RCTs with 297 participants comparing PUL to sham surgery or TURP. The mean age was 65.6 years and mean International Prostate Symptom Score was 22.7. Mean prostate volume was 42.2 mL. We considered review outcomes measured up to and including 12 months after randomization as short-term and later than 12 months as long-term. For patient-reported outcomes, lower scores indicate more urological symptom improvement and higher quality of life. In contrast, higher scores refers to better erectile and ejaculatory function.PUL versus sham: based on one study of 206 randomized participants with short follow-up (up to three months), PUL may lead to a clinically important improvement in urological symptom scores (mean difference (MD) -5.20, 95% confidence interval (CI) -7.44 to -2.96; low-certainty evidence) and likely improves quality of life (MD -1.20, 95% CI -1.67 to -0.73; moderate-certainty evidence). We are uncertain whether PUL increases major adverse events (very low-certainty evidence). There were no retreatments reported in either study group by three months. PUL likely results in little to no difference in erectile function (MD -1.40, 95% CI -3.24 to 0.44; moderate-certainty evidence) and ejaculatory function (MD 0.50, 95% CI -0.38 to 1.38; moderate-certainty evidence).PUL versus TURP: based on one study of 91 randomized participants with a short follow-up (up to 12 months), PUL may result in a substantially lesser improvement in urological symptom scores than TURP (MD 4.50, 95% CI 1.10 to 7.90; low-certainty evidence). PUL may result in a slightly reduced or similar quality of life (MD 0.30, 95% CI -0.49 to 1.09; low-certainty evidence). We are very uncertain whether PUL may cause fewer major adverse events but increased retreatments (both very low-certainty evidence). PUL probably results in little to no difference in erectile function (MD 0.80, 95% CI -1.50 to 3.10; moderate-certainty evidence), but probably results in substantially better ejaculatory function (MD 5.00, 95% CI 3.08 to 6.92; moderate-certainty evidence).With regards to longer term follow-up (up to 24 months) based on one study of 91 randomized participants, PUL may result in a substantially lesser improvement in urological symptom score (MD 6.10, 95% CI 2.16 to 10.04; low-certainty evidence) and result in little worse to no difference in quality of life (MD 0.80, 95% CI 0.00 to 1.60; low-certainty evidence). The study did not report on major adverse events. We are very uncertain whether PUL increases retreatment (very low-certainty evidence). PUL likely results in little to no difference in erectile function (MD 1.60, 95% CI -0.80 to 4.00; moderate-certainty evidence), but may result in substantially better ejaculatory function (MD 4.30, 95% CI 2.17 to 6.43; low-certainty evidence).We were unable to perform any of the predefined secondary analyses for either comparison.We found no evidence for other comparisons such as PUL versus laser ablation or enucleation. PUL appears less effective than TURP in improving urological symptoms both short-term and long term, while quality of life outcomes may be similar. The effect on erectile function appears similar but ejaculatory function may be better. We are uncertain about major adverse events short-term and found no long-term information. We are very uncertain about retreatment rates both short-term and long-term. We were unable to assess the effects of PUL in subgroups based on age, prostate size, or symptom severity and also could not assess how PUL compared to other surgical management approaches. Given the large numbers of alternative treatment modalities to treat men with LUTS secondary to BPH, this represents important information that should be shared with men considering surgical treatment.


Solution structure and sugar-binding mechanism of mouse latrophilin-1 RBL: a 7TM receptor-attached lectin-like domain.
Latrophilin-1 (Lat-1), a target receptor for alpha-Latrotoxin, is a putative G protein-coupled receptor implicated in synaptic function. The extracellular portion of Lat-1 contains a rhamnose binding lectin (RBL)-like domain of unknown structure. RBL domains, first isolated from the eggs of marine species, are also found in the ectodomains of other metazoan transmembrane proteins, including a recently discovered coreceptor of the neuronal axon guidance molecule SLT-1/Slit. Here, we describe a structure of this domain from the mouse Lat-1. RBL adopts a unique alpha/beta fold with long structured loops important for monosaccharide recognition, as shown in the structure of a complex with L-rhamnose. Sequence alignments and mutagenesis show that residues important for carbohydrate binding are often absent in other receptor-attached examples of RBL, including the SLT-1/Slit coreceptor. We postulate that this domain class facilitates direct protein-protein interactions in many transmembrane receptors.


Chronic vagal nerve stimulation has no effect on tachycardia-induced heart failure progression or excitation-contraction coupling.
Autonomic dysregulation plays a key role in the development and progression of heart failure (HF). Vagal nerve stimulation (VNS) may be a promising therapeutic approach. However, the outcomes from clinical trials evaluating VNS in HF have been mixed, and the mechanisms underlying this treatment remain poorly understood. Intermittent high-frequency VNS (pulse width 300 µs, 30 Hz stimulation, 30 s on, and 300 s off) was used in healthy sheep and sheep in which established HF had been induced by 4 weeks rapid ventricular pacing to assess (a) the effects of VNS on intrinsic cardiac vagal tone, (b) whether VNS delays the progression of established HF, and (c) whether high-frequency VNS affects the regulation of cardiomyocyte calcium handling in health and disease. VNS had no effect on resting heart rate or intrinsic vagal tone in the healthy heart. Although fewer VNS-treated animals showed subjective signs of heart failure at 6 weeks, overall VNS did not slow the progression of clinical or echocardiographic signs of HF. Chronic VNS did not affect left ventricular cardiomyocyte calcium handling in healthy sheep. Rapid ventricular pacing decreased the L-type calcium current and calcium transient amplitude, but chronic VNS did not rescue dysfunctional calcium handling. Overall, high-frequency VNS did not prevent progression of established HF or influence cellular excitation-contraction coupling. However, a different model of HF or selection of different stimulation parameters may have yielded different results. These results highlight the need for greater insight into VNS dosing and parameter selection and a deeper understanding of its physiological effects.


APO-1-induced apoptosis of leukemia cells from patients with adult T-cell leukemia.
The 48-Kd cell-surface protein APO-1 is a new member of the nerve growth factor (NGF)/tumor necrosis factor (TNF) receptor superfamily. APO-1 is expressed on various cells, including activated T and B cells and some lymphoid and nonlymphoid cell lines. Triggering of APO-1 by the monoclonal antibody anti-APO-1 induces programmed cell death (apoptosis) in APO-1-expressing cells. APO-1 is also present on T-cell lines derived from patients with adult T-cell leukemia (ATL). Therefore, we investigated APO-1 expression and APO-1-mediated induction of apoptosis ex vivo in cells from patients with ATL. Fresh leukemic cells from nine patients with ATL were assayed for APO-1 expression by two-color immunofluorescence. The leukemic cells from all patients strongly expressed APO-1. Incubation of ATL cells with anti-APO-1 in vitro inhibited spontaneous and cytokine-mediated DNA synthesis. Furthermore, DNA isolated from cells treated with anti-APO-1 exhibited polynucleosomal DNA fragmentation (DNA ladder) characteristic for apoptotic cell death. The analysis of APO-1-mediated apoptosis may represent a new approach to the study of growth control in lymphoid malignancies. In addition, induction of apoptosis by administration of anti-APO-1 may represent a new therapeutic approach for aggressive T-cell malignancies such as ATL.


Monkeypox (Mpox) requires continued surveillance, vaccines, therapeutics and mitigating strategies.
The widespread outbreak of the monkeypox virus (MPXV) recognized in 2022 poses new challenges for public healthcare systems worldwide. With more than 86,000 people infected, there is concern that MPXV may become endemic outside of its original geographical area leading to repeated human spillover infections or continue to be spread person-to-person. Fortunately, classical public health measures (e.g., isolation, contact tracing and quarantine) and vaccination have blunted the spread of the virus, but cases are continuing to be reported in 28 countries in March 2023. We describe here the vaccines and drugs available for the prevention and treatment of MPXV infections. However, although their efficacy against monkeypox (mpox) has been established in animal models, little is known about their efficacy in the current outbreak setting. The continuing opportunity for transmission raises concerns about the potential for evolution of the virus and for expansion beyond the current risk groups. The priorities for action are clear: 1) more data on the efficacy of vaccines and drugs in infected humans must be gathered; 2) global collaborations are necessary to ensure that government authorities work with the private sector in developed and low and middle income countries (LMICs) to provide the availability of treatments and vaccines, especially in historically endemic/enzootic areas; 3) diagnostic and surveillance capacity must be increased to identify areas and populations where the virus is present and may seed resurgence; 4) those at high risk of severe outcomes (e.g., immunocompromised, untreated HIV, pregnant women, and inflammatory skin conditions) must be informed of the risk of infection and be protected from community transmission of MPXV; 5) engagement with the hardest hit communities in a non-stigmatizing way is needed to increase the understanding and acceptance of public health measures; and 6) repositories of monkeypox clinical samples, including blood, fluids, tissues and lesion material must be established for researchers. This MPXV outbreak is a warning that pandemic preparedness plans need additional coordination and resources. We must prepare for continuing transmission, resurgence, and repeated spillovers of MPXV.


Characterization of sheared colloidal aggregation using Langevin dynamics simulation.
Aggregation of colloidal particles under shear is studied in model systems using a Langevin dynamics model with an improved interparticle interaction potential. In the absence of shear, aggregates that form are characterized by compact structure at small scales and ramified structure at larger scales. This confirms the structural crossover mechanism previously suggested by Sorensen and coworkers, that colloidal aggregation occurs due to monomer addition at small scales and due to cluster-cluster aggregation at large scales. The fractal dimension of nonsheared aggregates is scale-dependent. Smaller aggregates have a higher fractal dimension than larger ones, but the radius of gyration where this crossover occurs is independent of potential well depth for sufficiently deep wells. When these aggregates are subjected to shear they become anisotropic and form extended cigar-like structures. The size of sheared anisotropic aggregates in the direction perpendicular to the shear flow is limited by shear-induced breakage because the shear force dominates interparticle attraction for sufficiently large aggregates. Anisotropic aggregates are not completely characterized by a single radius of gyration, but rather by an inertia ellipsoid. Consequently the fractal dimension is no longer an adequate metric to properly characterize them, and to identify changes in their structure from their nonsheared isotropic counterparts. We introduce a new compactness-anisotropy analysis that characterizes the structure of anisotropic aggregates and allows us to distinguish between aggregates from sheared and nonsheared systems. Finally, using the ratio of interparticle force to the shear force f_{pot,sh} we are able to characterize different outcomes of sheared aggregation as a function of dimensionless well depth and Péclet number.


Prevalence of neurologic autoantibodies in cohorts of patients with new and established epilepsy.
Autoantibodies to specific neurologic proteins are associated with subacute onset encephalopathies, which often present with seizures that are poorly controlled by conventional antiepileptic drugs (AEDs). Previous cross-sectional studies have found specific neurologic antibodies in a small proportion of people with established epilepsy, but these investigations have seldom included patients with recent diagnosis. We screened two large epilepsy cohorts to investigate the prevalence of multiple autoantibodies in adult patients with either established or newly diagnosed, untreated epilepsy. Eleven percent of patients had antibodies to one or more antigen: voltage-gated potassium channel (VGKC) complex proteins (5%), glycine receptors (3%), and glutamic acid decarboxylase (GAD) and N-methyl-D-aspartate (NMDA) receptors (1.7% each). There was no difference in the prevalence of antibodies, individually or collectively, between patients with established and newly diagnosed epilepsy or with generalized or focal epilepsy. There was, however, a significantly higher prevalence of positive antibody titers in patients with focal epilepsy of unknown cause than in those with structural/metabolic focal epilepsy (14.8% vs. 6.3%; p < 0.02). Newly diagnosed antibody-positive patients were less likely to achieve adequate seizure control with initial treatment than antibody-negative patients, but this difference failed to reach statistical significance. The presence of autoantibodies is equally common in newly diagnosed and established epilepsy, it is therefore unlikely to be an epiphenomenon of long-standing refractory seizures.