Mitochondria as signaling organelles.
Almost 20 years ago, the discovery that mitochondrial release of cytochrome c initiates a cascade that leads to cell death brought about a wholesale change in how cell biologists think of mitochondria. Formerly viewed as sites of biosynthesis and bioenergy production, these double membrane organelles could now be thought of as regulators of signal transduction. Within a few years, multiple other mitochondria-centric signaling mechanisms have been proposed, including release of reactive oxygen species and the scaffolding of signaling complexes on the outer mitochondrial membrane. It has also been shown that mitochondrial dysfunction causes induction of stress responses, bolstering the idea that mitochondria communicate their fitness to the rest of the cell. In the past decade, multiple new modes of mitochondrial signaling have been discovered. These include the release of metabolites, mitochondrial motility and dynamics, and interaction with other organelles such as endoplasmic reticulum in regulating signaling. Collectively these studies have established that mitochondria-dependent signaling has diverse physiological and pathophysiological outcomes. This review is a brief account of recent work in mitochondria-dependent signaling in the historical framework of the early studies.


Satellite radiation measurements and clear air turbulence probability.
Radiance gradients determined from data gathered by the infrared spectrometers aboard the Nimbus III and Nimbus IV satellites were related to the probabilities of clear air turbulence, as inferred from regular pilot reports. Such radiance gradients represent rather large-scale vertical wind shear. Clear air turbulence is least likely in regions where the radiance gradient is small. The results of this exploratory study show that satellite data can be used to design flight paths so that the probability of encountering clear air turbulence is extremely small.


CD14 and toll-like receptors: potential contribution of genetic factors and mechanisms to inflammation and allergy.
Innate and adaptive immune responses evolve as protective mechanisms against infectious microorganisms in humans. CD14 and toll-like receptors (TLRs) are examples of pattern recognition receptors that detect antigenic molecules on the surface of gram-positive (peptidoglycans, lipoteichoic acid) and gram-negative (lipopolysaccharide) bacteria. In vitro studies suggest that lipopolysaccharide is a potent inducer of interleukin-12 production that is mediated by both CD14 and TLR4. The associated increase in interferon-gamma steers our immune system away form the allergy-driven type-2 helper T cell phenotype. Epidemiological studies that shed light on the possible protective influences of natural microbial exposure on asthma and atopy development will be discussed. Recent insights into the complex mechanisms of human innate immunity suggest that genetic variability in genes encoding its components may alter the susceptibility to develop atopic disorders and other complex human diseases. The findings of these genetic association studies will be presented. Although highly conserved across a wide range of species, innate immunity genes demonstrate considerable inter-ethnic variability predominantly in the form of single nucleotide polymorphisms. The frequencies of these polymorphisms in CD14 and TLR genes in different ethnic groups will be discussed. Genetic variation in these genes may also play a role in the development of other human diseases that have an inflammatory component. Lastly, the prospect of using immunomodulatory agents targeting on the innate immunity to treat or even prevent asthma and other allergic diseases will be discussed.


Rainbow trout leucocyte activity: influence on the ectoparasitic monogenean Gyrodactylus derjavini.
The ectoparasitic monogenean Gyrodactylus derjavini from rainbow trout Oncorhynchus mykiss was exposed in vitro to macrophages isolated as peritoneal exudate cells or as pronephros cells from the host. Cells colonized the parasite especially in the mannose-rich regions in the cephalic ducts where ciliated structures were abundant. Opsonization with fresh serum, in contrast to heat-inactivated serum, enhanced colonization also on other body parts. The adverse effect of the activated macrophages towards G. derjavini was associated with a heat-labile component released from these cells to the culture medium. Analysis of substances released from the cells showed reactivity for a number of enzymes, complement factor C3, interleukin (Il-1) and reactive oxygen metabolites. Chemotaxis assays with pronephric leucocytes showed chemoattractants in G. derjavini, and the respiratory burst level of macrophages was slightly elevated due to parasite exposure. It is suggested that skin leucocytes contribute to an increased level of complement factors in the trout skin during the host response, whereby a hostile microenvironment for the parasites is created. In addition, the IL-1 production could affect mucous cell secretion and hyperplasia and add to the antiparasitic action of the epithelium. Likewise, reactive oxygen metabolites and various enzymes are likely to be involved in the skin response.


Comparison of the effectiveness of the Hanen Parent Programme and traditional clinic therapy.
Both direct (clinician to child) and indirect (clinician to carer) approaches are currently used in the management of children with language delay, but there is as yet little evidence about their relative effects or resource implications. This research project compared the Hanen Parent Programme (HPP) in terms of its effectiveness and consequent suitability for an inner-city UK population with clinic-based, direct intervention. Thirty-seven children aged 2;06-3;06 years with a diagnosis of language impairment and their parents took part in the research project. The children were allocated on a geographical basis to receive therapy either as part of an HPP or in a clinic. Nineteen children and their families took part in one of the five Hanen groups that ran successively over 16 months; 18 children and their families received clinic-based intervention. The children's language was assessed using the PLS-3 (UK version) and from an analysis of audio-taped parent and child interaction at three assessment points, one pretherapy and two post-therapy over 12 months. Two parent language measures were also analysed. Significant gains in language scores were shown by 71% of the children over 12 months. There were no statistically significant differences in child language scores between the two therapy groups at any assessment point. However, the HPP was twice as intensive (in terms of therapist time) as clinic therapy based on average group size, which has resource implications. CONCLUSIONS. Results suggest that there are parent and child factors that need consideration when choosing an appropriate intervention programme for a child with language impairment. Parental expectations, existing interaction style and the level of child language may be important influencing factors.


End joining at Caenorhabditis elegans telomeres.
Critically shortened telomeres can be subjected to DNA repair events that generate end-to-end chromosome fusions. The resulting dicentric chromosomes can enter breakage-fusion-bridge cycles, thereby impeding elucidation of the structures of the initial fusion events and a mechanistic understanding of their genesis. Current models for the molecular basis of fusion of critically shortened, uncapped telomeres rely on PCR assays that typically capture fusion breakpoints created by direct ligation of chromosome ends. Here we use independent approaches that rely on distinctive features of Caenorhabditis elegans to study the frequency of direct end-to-end chromosome fusion in telomerase mutants: (1) holocentric chromosomes that allow for genetic isolation of stable end-to-end fusions and (2) unique subtelomeric sequences that allow for thorough PCR analysis of samples of genomic DNA harboring multiple end-to-end fusions. Surprisingly, only a minority of end-to-end fusion events resulted from direct end joining with no additional genome rearrangements. We also demonstrate that deficiency for the C. elegans Ku DNA repair heterodimer does not affect telomere length or cause synthetic effects in the absence of telomerase.


Mitochondrial DNA and Y-chromosomal diversity in ancient populations of domestic sheep (Ovis aries) in Finland: comparison with contemporary sheep breeds.
Several molecular and population genetic studies have focused on the native sheep breeds of Finland. In this work, we investigated their ancestral sheep populations from Iron Age, Medieval and Post-Medieval periods by sequencing a partial mitochondrial DNA D-loop and the 5'-promoter region of the SRY gene. We compared the maternal (mitochondrial DNA haplotypes) and paternal (SNP oY1) genetic diversity of ancient sheep in Finland with modern domestic sheep populations in Europe and Asia to study temporal changes in genetic variation and affinities between ancient and modern populations. A 523-bp mitochondrial DNA sequence was successfully amplified for 26 of 36 sheep ancient samples i.e. five, seven and 14 samples representative of Iron Age, Medieval and Post-Medieval sheep, respectively. Genetic diversity was analyzed within the cohorts. This ancient dataset was compared with present-day data consisting of 94 animals from 10 contemporary European breeds and with GenBank DNA sequence data to carry out a haplotype sharing analysis. Among the 18 ancient mitochondrial DNA haplotypes identified, 14 were present in the modern breeds. Ancient haplotypes were assigned to the highly divergent ovine haplogroups A and B, haplogroup B being the major lineage within the cohorts. Only two haplotypes were detected in the Iron Age samples, while the genetic diversity of the Medieval and Post-Medieval cohorts was higher. For three of the ancient DNA samples, Y-chromosome SRY gene sequences were amplified indicating that they originated from rams. The SRY gene of these three ancient ram samples contained SNP G-oY1, which is frequent in modern north-European sheep breeds. Our study did not reveal any sign of major population replacement of native sheep in Finland since the Iron Age. Variations in the availability of archaeological remains may explain differences in genetic diversity estimates and patterns within the cohorts rather than demographic events that occurred in the past. Our ancient DNA results fit well with the genetic context of domestic sheep as determined by analyses of modern north-European sheep breeds.


Resisting, Recognizing, and Returning: A Three-Component Model and Review of Persistence in Episodic Goals.
According to prior work, persistent goal pursuit is a continuous process where persisting is a matter of resisting the urge to give up. In everyday goals, however, persistence is often episodic, and its causes are more complex. People pause and resume pursuit many times. Whether people persist reflects more than will power and motivation, it also reflects the other goals they pursue, their resources, and the attentional demands of daily life. People can fail to persist not just because they gave up, but also because they failed to act. We propose a general model of persistence that accommodates the complexity of episodic goals. We argue that persistent goal pursuit is a function of three processes: resisting the urge to give up, recognizing opportunities for pursuit, and returning to pursuit. The broad factors that help and hurt persistence can be organized within these components. These components can also explain the mechanisms of four effective strategies for persistence: removing distractions, using reminders, using implementation intentions, and forming habits. The recognizing-resisting-returning model integrates and improves on extant theories of persistence and goal pursuit and is consistent with empirical work from laboratory and naturalistic settings.


Radiographic and Clinical Outcomes of the Shoulder in Long-Term Follow-Up of Brachial Plexus Birth Injury.
The purpose of this study was to examine radiographic and functional outcomes of the shoulder in brachial plexus birth injury (BPBI) patients at long-term follow-up. Medical records were reviewed at the Shriners Hospital in St. Louis to identify patients older than 18 years with a diagnosis of BPBI. Twenty-five patients were included in this study. Evaluations included radiographic examination, clinical examination, and outcome measures including the Mallet classification, visual analog scale (VAS) for shoulder pain, American Shoulder and Elbow Shoulder score (ASES), quick Disabilities of the Arm, Shoulder, and Hand (QuickDASH), Simple Shoulder Test (SST), and Patient-Reported Outcomes Measurement Information System (PROMIS) domain measures. The mean age of the patients included was 44 years. Average follow-up was 37 years. No patient had primary nerve reconstruction surgery. Thirteen patients had a history of shoulder surgery at an average age of 6 years. There was a wide spectrum of deformity seen radiographically. Fifty-five percent of patients had evidence of glenoid retroversion, posterior humeral translation, or a biconcave glenoid. An additional 20% of patients had complete posterior dislocation of the humeral head. Despite the degenerative changes seen on radiographs, the mean VAS for pain was low, at 2. The mean ASES shoulder score was 54. The mean QuickDASH score was 30. Mean PROMIS Physical Function, Pain Interference, Depression, and Anxiety scores were near the means for the general population, with similar outcomes in patients treated with shoulder surgery compared with those treated without surgery. Adult BPBI patients experience minimal shoulder pain despite deformity and degenerative changes on shoulder radiographs. Whereas there are significant deficits in range of motion and function, PROMIS scores in adult BPBI patients are normal. Therapeutic IV.


Monogamy as a Barrier to Human Papillomavirus Catch-Up Vaccination.
Background: Human papillomavirus (HPV) is the most prevalent sexually transmitted infection (STI) in the United States. Although a vaccine to prevent HPV infection exists, only 53.7% of females 13-17 years of age were up-to-date on the HPV vaccination series in 2018. There is a catch-up period of vaccination for females 18-26 years of age that shows consistent underparticipation. A potential barrier to vaccination is relationship status, as long-term relationships may negatively impact HPV risk perception. This study examined monogamy as a risk factor for nonvaccination and explored how risk perception may influence this association. Materials and Methods: An electronic survey was distributed to females 18-26 years of age who attended a large public university in the mid-Atlantic region (n = 629). Multivariable and descriptive statistics were estimated using SAS 9.4 to explore the likelihood of vaccination during the catch-up period by relationship status. Results: Most participants had received the HPV vaccine, a small proportion of whom received it during the catch-up period. After adjusting for confounders, women who were in monogamous relationships were significantly less likely to have participated in HPV catch-up vaccination compared to women who were single and dating (adjusted odds ratio: 0.36, 95% confidence interval: 0.15, 0.87). Women in monogamous relationships had a lower average sexually transmitted disease (STD) risk perception compared to women who were single and dating (p < 0.0001). Conclusions: A decreased risk perception may present a barrier to participating in catch-up vaccination for monogamous women. Practitioners and the public health community should focus on communicating HPV risk to women in monogamous relationships, especially given the recently expanded age range for HPV vaccination.


The foot-and-mouth disease epidemic in Dumfries and Galloway, 2001. 2: Serosurveillance, and efficiency and effectiveness of control procedures after the national ban on animal movements.
After the foot-and-mouth disease (FMD) epidemic in Dumfries and Galloway in south-west Scotland in 2001, serosurveillance of sheep remaining in the 3 km radius Protection Zones around Infected Premises (IPS), and within a 10 km radius of IPS, revealed no evidence of infection. The epidemic was brought under control by a range of traditional techniques: slaughter of all animals on IPS and of veterinary-assessed Dangerous Contacts (DCS), movement restrictions, biosecurity, tracing of potential sources and spread of virus, and surveillance of At-Risk premises. Novel pre-emptive slaughter of FMD-susceptible animals on premises contiguous to IPS, and small ruminants and pigs on premises within 3 km of IPSs, commenced after the epidemic had peaked. Most of the traditional control procedures were undertaken quickly and with appropriate priority. Animals on IPS were usually slaughtered within one day of confirmation, and veterinary-assessed DCS within two days of confirmation of relevant IPS (a median of two days). The pre-emptive contiguous and 3 km culls took somewhat longer (medians of five and 17 days, respectively). IPS were most commonly identified as a result of reporting by farmers or their veterinarians (72 per cent of IPS); veterinary clinical patrols identified 16 per cent, while veterinary assessment of DCS and tracing each identified 5 per cent. No evidence of infection was found on any pre-emptively contiguously culled premises, and IPS were declared only on three 3 km cull premises. The time from estimated first lesion to end of slaughter on an IP was found, by regression analysis, to be a key component in effective control, manifested by a reduction in the estimated dissemination rate (EDR); there was little evidence that the intensity of contiguous culling affected the EDR. Patrols and serological surveillance of residual animals within 10 km of IPS, supported by more extensive evidence from elsewhere in the UK, suggested that cryptic infection in sheep was not widespread. Ultimately, there was insufficient evidence to support the effectiveness of 3 km pre-emptive culling as a control procedure.


Establishing key components of yoga interventions for musculoskeletal conditions: a Delphi survey.
Evidence suggests yoga is a safe and effective intervention for the management of physical and psychosocial symptoms associated with musculoskeletal conditions. However, heterogeneity in the components and reporting of clinical yoga trials impedes both the generalization of study results and the replication of study protocols. The aim of this Delphi survey was to address these issues of heterogeneity, by developing a list of recommendations of key components for the design and reporting of yoga interventions for musculoskeletal conditions. Recognised experts involved in the design, conduct, and teaching of yoga for musculoskeletal conditions were identified from a systematic review, and invited to contribute to the Delphi survey. Forty-one of the 58 experts contacted, representing six countries, agreed to participate. A three-round Delphi was conducted via electronic surveys. Round 1 presented an open-ended question, allowing panellists to individually identify components they considered key to the design and reporting of yoga interventions for musculoskeletal conditions. Thematic analysis of Round 1 identified items for quantitative rating in Round 2; items not reaching consensus were forwarded to Round 3 for re-rating. Thirty-six panellists (36/41; 88%) completed the three rounds of the Delphi survey. Panellists provided 348 comments to the Round 1 question. These comments were reduced to 49 items, grouped under five themes, for rating in subsequent rounds. A priori group consensus of ≥80% was reached on 28 items related to five themes concerning defining the yoga intervention, types of yoga practices to include in an intervention, delivery of the yoga protocol, domains of outcome measures, and reporting of yoga interventions for musculoskeletal conditions. Additionally, a priori consensus of ≥50% was reached on five items relating to minimum values for intervention parameters. Expert consensus has provided a non-prescriptive reference list for the design and reporting of yoga interventions for musculoskeletal conditions. It is anticipated future research incorporating the Delphi guidelines will facilitate high quality international research in this field, increase homogeneity of intervention components and parameters, and enhance the comparison and reproducibility of research into the use of yoga for the management of musculoskeletal conditions.


Numerical matching during cerebellar development: quantitative analysis of granule cell death in staggerer mouse chimeras.
Cell death is a common yet puzzling feature of the development of many populations of neurons in the CNS. In the invertebrate phyla, such death is often preprogrammed; by contrast, in vertebrates, the best studied examples of histogenetic cell death are influenced by interactions among the neurons and their target. One attempt to explain this seemingly wasteful scheme of development has led to the hypothesis that this target-related cell death allows 2 populations of cells, which develop in isolation, to come into numerical and functional balance and hence to provide an epigenetic "buffer" mechanism to accommodate developmental variations. In the current study we have examined the extent to which the cell death observed in the cerebellar granule cell population serves to numerically match these neurons with their primary postsynaptic target, the Purkinje cell. Staggerer chimeras were made by aggregating 8-cell staggerer embryos with embryos of wild-type genotype. The cerebella of the resulting animals developed with widely varying numbers of normal (wild-type) Purkinje cell targets. Although staggerer Purkinje cells were present in the chimeric brains, these cells are intrinsically deficient in their normal developmental program (in the mutant, because of this deficiency, 100% of the granule cells die). Both granule cells and Purkinje cells were counted in chimeras and several wild-type mice. The results reveal that the number of granule cells present in these brains has a linear relationship with the number of Purkinje cells, and that the line connecting the points intersects the Y-axis close to the origin. These observations suggest that numerical matching is an important function of target-related cell death in the granule cell population.(ABSTRACT TRUNCATED AT 250 WORDS)


A Systematic Mixed Studies Review of the Intersections of Social-Ecological Factors and HIV Stigma in People Living With HIV in the U.S. South.
The role of stigma on psychological wellness and treatment outcomes in people living with HIV (PLWH) has been well documented. However, within the context of the southern United States, the intersection between HIV-related stigma and social-ecological factors has been understudied. Thus, a results-based convergent, mixed synthesis design was used to examine the manifestations of HIV-related stigma in PLWH in the U.S. South. A literature search was conducted using PsycINFO, PubMed (includes MEDLINE), and CINAHL. The first level of screening by title and abstract was administered on 1,829 articles. A full-text screening of 169 studies was completed, and a total of 30 relevant articles were extracted. The mixed synthesis highlighted intervention strategies that can reduce HIV-related stigma while promoting positive health-behavior change. The findings of this review underscored the uniqueness of PLWH in the south and demonstrated the crucial role of intersectionality in investigating HIV-related stigma in treating and preventing HIV.


Rheological properties of super critical CO2 with CuO: Multi-scale computational modeling.
A multi-scale computational methodology based on the density functional theory and molecular dynamics has been used to investigate the rheological properties of super critical CO2 with CuO nano-particle (NP). Density functional theory which treats the electron density as the central variable has been used to explore the adsorption of CO2 molecules on the two most stable CuO surfaces [i.e., (111) and (011)] at absolute zero. The results of this theory would provide valuable information to make CuO NPs with the surface where the CO2 adsorption is maximum in order to have a stronger mono-layer of adsorbed CO2 molecules on the surface of the NP which is the most crucial factor in formation of a stable nanofluid. The results show that the CO2 molecule is adsorbed more strongly on the (011) surface with an adsorption energy of -99.06 kJ/mol compared to the (111) surface. A computational methodology based on molecular dynamics has been used to evaluate the enhancement of the rheological properties of the super-critical CO2 liquid based nanofluid at different temperatures and pressures. In this scale, first, the CO2 liquid has been modeled by employing the condensed-phase optimized molecular potentials for atomistic simulation studies (COMPASS) force field potential and the fluid properties computed are in excellent agreement with the literature and experiment values. Second, the nanofluid has been modeled in order to study the enhancement of the fluid properties with the CuO NPs. The charged optimized many-body force field potential has been employed to consider the effect of the charge transferring between the NPs and liquid molecules and breaking of existing bonds and the formation of new bonds. The COMPASS force field potential is also employed for the interactions between CO2 molecules. The combination of these potentials is quite a new approach for the study of the super-critical (SC)-CO2 based nanofluid. The results show that the viscosity of the SC-CO2 is enhanced between 1.3 and 2.5 times under the temperature and pressure conditions studied.


Subperiosteal hematoma of the orbit associated with sinusitis.
Subperiosteal hematoma of the orbit is a rare but well-recognized entity, usually caused by trauma. Two cases of subperiosteal hematoma associated with sinusitis are presented. A 44-year-old woman experienced the sudden onset of proptosis, and decreased visual acuity. Computed tomographic scanning and magnetic resonance imaging revealed a frontoethmoidal mucocele and a biconvex mass in the upper part of the left orbit. The mucocele was drained during nasal endoscopic surgery and the subperiosteal hematoma was evacuated during superior orbitotomy. A 42-year-old man had a headache and proptosis. Computed tomographic scan revealed sinusitis and subperiosteal orbital hematoma of the left orbit. Subperiosteal orbital hematoma associated with sinusitis is extremely rare but should be suspected in a patient with acute onset of proptosis in whom computed tomographic scanning reveals paranasal sinusitis.


American Academy of Pediatrics. Committee on Adolescence. Contraception in adolescents.
The risks and negative consequences of adolescent sexual intercourse are of national concern, and promoting sexual abstinence is an important goal of the American Academy of Pediatrics. In previous publications, the American Academy of Pediatrics has addressed important issues of adolescent sexuality, pregnancy, sexually transmitted diseases, and contraception. The development of new contraceptive technologies mandates a revision of this policy statement, which provides the pediatrician with an updated review of adolescent sexuality and use of contraception by adolescents and presents current guidelines for counseling adolescents on sexual activity and contraceptive methods.


Local variability of Arctic mesozooplankton biomass and production: A case summer study.
The Barents Sea is a highly productive ecosystem within the Arctic Ocean. The overall biological productivity in this region relies heavily on the secondary mesozooplankton production (MZP). Previous research has primarily focused on mesozooplankton abundance (MZA) and biomass (MZB), lacking a comprehensive analysis of the environmental factors that influence MZP in the Arctic marine environment. The primary objective of this study was to examine the key factors responsible for the spatial variability in the community structure and MZP during the summer season. Data were collected from 52 stations in the central Barents Sea, specifically during post-bloom conditions in 2015. Through cluster analysis, two distinct groups of stations were identified, differing in terms of mesozooplankton taxa abundance. Copepods were found to dominate the mesozooplankton assemblages, comprising 89% of total MZA, 83% of MZB, and 68% of MZP. The biomass stocks in the study area varied from 8 to 102 mg dry mass (DM) m-3, with an average of 44 mg DM m-3. MZP rates ranged from 0.34 to 2.33 mg DM m-3 day-1, with an average of 1.16 mg DM m-3 day-1. The highest MZB and MZP values were observed at frontal zones that separated relatively warm and cold waters. Through redundancy analysis, it was determined that the primary environmental factors affected the distribution of mesozooplankton were longitude, latitude, and sampling depth. Temperature, salinity, and chlorophyll a concentration were found to have a less significant impact. These findings emphasize the importance of oceanographic conditions as the main predictors of mesozooplankton distribution during the summer season in the Arctic marine environment. This study highlights the essential role of environmental forcing in determining the productivity of Arctic marine zooplankton. Given the ongoing climatic changes, the results of this report can serve as a valuable tool for monitoring pelagic ecosystems in the Arctic.


Effects of thymectomy on bone growth in the rat.
The effects of thymectomy (TMX) at 1 or 5 days of age on the subsequent growth of bone have been determined in male and female Holtzman rats. The rats were maintained under aseptic conditions to minimize any effects of infection or wasting disease on bone growth. Femur length and distal epiphyseal cross sectional area were significantly reduced in TMX female rats at 3 months of age. At 1 month of age, the width of the undifferentiated zone was increased and at all subsequent times the columnar zone and growth plate were narrower in TMX rats. The incorporation of 35S into the femur, particularly the proximal and distal growth plates, was uniformly reduced in TMX rats as determined by scintillation counting and autoradiography. Additionally, there was an obvious overall reduction in alcian blue stationing intensity in the growth plates of TMX rats when compared to sham-operated controls. These results suggest a significant reduction in detectable glycosaminoglycan (GAG) content and in bone growth in TMX rats. Although there were minor variations, there were no major differences attributable to sex or timing of thymectomy. At present the causes of this moderate reduction in bone growth in TMX rats are unknown. One explanation might be that the thymus supplies or is in some other way involved in the provision of a hormone and/or cells that are required for normal bone growth. Another possibility is that the thymus is normally a source of immune suppressor cells which are needed to limit auto-immune reactivity against GAG or other antigenic determinants associated with bone. Thymectomy would remove these suppressor cells, thus permitting an auto-immune response which interferes with bone growth. Finally, perhaps the most plausible explanation is that thymectomy reduces cell production in the bone marrow which, in particular, reduces the turnover of monocytes. This reduced output of monocytes in TMX rats might ultimately lead to a reduction in osteoclast activity necessary for normal remodelling and growth of bone. We currently favour this last explanation.


[Treatment of malaria: outlook].
Prospects for new and effective antimalarial treatments have improved significantly in the past few years. There are more funds, new operational paradigms and actors, and more products in the pipeline. However, the limited novelty of the products currently in clinical development means that more emphasis therefore needs to be placed on the discovery of novel chemical entities to achieve effective, sustainable control. The development and deployment of new products is both lengthy and complex.


[Indirect calorimetry in critical ill patients: validity of measurement for ten minutes].
There are no gold standards on the duration and frequency of the measurement of indirect calorimetry, a fact of importance in daily clinical practice. An assessment of is made of the degree of concordance between energy expenditure at rest (EER) measured over a short interval (10 minutes) versus another prolonged measurement (1 hour). Sixty critically-ill patients, under sedation and analgesia with connection to mechanical ventilation, were studied. EER values were determined by means of a metabolic computer analysis (Engström Eliza) at rest. The reproducibility and the degree of concordance were assessed in the measurements made with both periods. The mean values of the EER determinations at 10 and 60 minutes were 1,818 +/- 319 kilocalories/day and 1,815 +/- 318 Kcal/day. The limits of the concordance between both times were -101 and +117 kilocalories/day and the correlation was significant (r = 0.98, p < 0.0001). In critically-ill patients under sedation and with mechanical ventilation, the measurement of EER may be taken over short periods of time (10 minutes) providing that baseline examination conditions are met, thus giving greater availability of the resources used to study indirect calorimetry.


Macular pigment and visual performance under glare conditions.
Many parameters of visual performance (e.g., contrast sensitivity) are compromised under glaring light conditions. Recent data indicate that macular pigment (MP) is strongly related to improvements in glare disability and photostress recovery based on a filtering mechanism. In this study, we assessed the causality of this relation by supplementing lutein and zeaxanthin for 6 months while measuring MP, glare disability, and photostress recovery. Forty healthy subjects (mean age = 23.9) participated in the study. Subjects were followed for 6 months and assessed at baseline, 1, 2, 4, and 6 months. Spatial density profiles of MP were measured using heterochromatic flicker photometry. Disability glare was measured using a 1 degree-diameter circular grating surrounded by a broadband glare source (a xenon-white annulus). The intensity of the annulus (11 degree inner and 12 degree outer diameters) was adjusted by the subject until the grating target was no longer seen. For the photostress recovery experiment, the time required to detect a 1 degree-diameter grating stimulus after a 5-s exposure to a 2.5 muW/cm2, 5 degree-diameter disk was recorded. Subjects were tested under central viewing and eccentric viewing (10 degree temporal retina) conditions. At the baseline time point, MP optical density (OD) at 30' eccentricity ranged from 0.08 to 1.04, and was strongly correlated with improved visual performance in the two glare tasks. After 6 months of lutein (L) and zeaxanthin (Z) supplementation, average MPOD (at 30' eccentricity) had increased from 0.41 to 0.57, and was shown to significantly reduce the deleterious effects of glare for both the visual performance tasks assessed. MP is strongly related to improvements in glare disability and photostress recovery in a manner strongly consistent with its spectral absorption and spatial profile. Four to 6 months of 12 mg daily L + Z supplementation significantly increases MPOD and improves visual performance in glare for most subjects.


Redesigning Work With a Lightweight Approach to Coordination Technology.
Coordination of information and resources is central to efficient ward management. Tools to aid coordination include standardized procedures and paper lists and, more recently, electronic tools such as whiteboards and mobile solutions. This study reports on the experiences and effects of designing and implementing collaboration technology, and how the use of light-weight technology has enabled redesign of traditional work processes. The process followed the principles for user-centered development and is evaluated through observations and informal interviews of users and stakeholders. In addition to the expected changes in work processes, we found that short redesign cycles with heavy user involvement afforded by this technology enabled implementation of new ways of structuring and combining information that were not foreseen. This turned the collaboration technology into a tool that also guided prioritization between tasks and contributed to the effects seen in use of resources, user satisfaction, and quality of treatment.


Analyzing the Effects of Psychotherapy on Weight Loss after Laparoscopic Gastric Bypass or Laparoscopic Adjustable Gastric Banding in Patients with Borderline Personality Disorder: A Prospective Study.
Personality disorders are frequently associated with eating disorders in obese patients and may negatively affect weight loss and maintenance after bariatric surgery. This non-randomized study aimed to assess the effects of different psychotherapeutic interventions on weight loss in a sample of patients with borderline personality disorder who underwent laparoscopic gastric bypass or laparoscopic adjustable gastric banding. A total of 153 bariatric patients meeting borderline personality disorder criteria were chosen voluntarily and consecutively to undergo an interpersonal individual treatment (n = 50), a dialectical behavioral group treatment (n = 50), or treatment as usual (n = 53) for a year after surgery. Their body mass index was measured before and at the end of each treatment. A total of 12 patients (7.8%) dropped out of the study. Significantly higher body mass index reductions were registered in both experimental groups (-14.2 and -9.4 kg/m2, respectively) compared with the treatment as usual group (-2.1 kg/m2; p < 0.01). Treated patients who underwent laparoscopic gastric bypass (total n = 94) showed better outcomes than those who underwent laparoscopic adjustable gastric banding (total n = 45), while no differences were observed in untreated patients. This study demonstrates the role of pre-operative psychological assessment and post-operative psychotherapeutic support in improving weight loss among bariatric patients with borderline personality disorder. A randomized controlled trial is needed to confirm these preliminary findings.


Maitotoxin-evoked gamma-aminobutyric acid release is due not only to the opening of calcium channels.
The effects of maitotoxin (MTX) on endogenous amino acid release were tested on highly purified striatal neurons differentiated in primary culture. MTX induced a large and concentration-dependent release of gamma-aminobutyric acid (GABA). This effect was abolished when experiments were performed in the absence of external Ca2+, and restored when Ca2+ ions were added after removing the MTX-containing Ca2+-free solution. MTX-induced amino acid release was not affected by 1 microM nifedipine and only slightly inhibited by 1 mM Co2+. MTX also induced a massive accumulation of 45Ca2+ in the neurons which, in contrast to the MTX-evoked GABA release, was totally blocked in the presence of 1 mM Co2+. Whereas 500 nM tetrodotoxin was without significant effect, MTX-evoked GABA release was dependent on the presence of external Na+ and sensitive to nipecotic acid, a GABA uptake inhibitor. It is concluded that, on striatal neurons, MTX induced Na+ influx only in the presence of external Ca2+. The increase in cytoplasmic Na+ ions then triggers the release of GABA.


HHLA2 Immune Checkpoint Is a Novel Prognostic Predictor in Hepatocellular Carcinoma.
Immunotherapy targeting PD-1/PD-L1 had low response rates and limited overall survival benefits in patients with hepatocellular carcinoma (HCC). We characterized the expression pattern and clinical significance of B7 family immune checkpoint proteins HHLA2, PD-L1, and B7-H4 in HCC. Immunohistochemistry for HHLA2, PD-L1, B7-H4, CD3, and CD8 was performed on tissue microarray slides from 103 surgically resected HCC specimens. Positive HHLA2 was detected in 63 (61.2%) cases, with 50 cases showing low expression and 13 cases showing high expression. Positive PD-L1 (combined positive score [CPS] ≥1) was detected in 27 (26.2%) cases, with the majority showing low expression (CPS <10) (n = 25) and only 2 cases showing high expression (CPS ≥10). The coexpression of PD-L1 and HHLA2 was observed in 17 (16.5%) cases. B7-H4 expression was detected in only one (1.0%) case. HHLA2-positive cases had significantly better survival than HHLA2-negative cases (P = .042). Positive HHLA2 correlated with higher density of CD8+ intratumoral lymphocytes (P = .015). The subgroup with both HHLA2 expression and high CD8+ cell density exhibited the most favorable prognosis (P = .036). HHLA2 is frequently expressed in HCC. Positive HHLA2 correlates with higher CD8+ cell density and favorable prognosis. HHLA2 may be considered a potential therapeutic immune target in HCC.


Nutrient sensing kinases PKA and Sch9 phosphorylate the catalytic domain of the ubiquitin-conjugating enzyme Cdc34.
Cell division is controlled in part by the timely activation of the CDK, Cdc28, through its association with G1 and G2 cyclins. Cdc28 complexes are regulated in turn by the ubiquitin conjugating enzyme Cdc34 and SCF ubiquitin ligase complexes of the ubiquitin-proteasome system (UPS) to control the initiation of DNA replication. Here we demonstrate that the nutrient sensing kinases PKA and Sch9 phosphorylate S97 of Cdc34. S97 is conserved across species and restricted to the catalytic domain of Cdc34/Ubc7-like E2s. Cdc34-S97 phosphorylation is cell cycle regulated, elevated during active cell growth and division and decreased during cell cycle arrest. Cell growth and cell division are orchestrated to maintain cell size homeostasis over a wide range of nutrient conditions. Cells monitor changes in their environment through nutrient sensing protein kinases. Thus Cdc34 phosphorylation by PKA and Sch9 provides a direct tether between G1 cell division events and cell growth.


Methylation silencing of angiopoietin-like 4 in rat and human mammary carcinomas.
Aberrant DNA methylation is deeply involved in the development and progression of human breast cancers, but its inducers and molecular mechanisms are still unclear. To reveal such inducers and clarify the molecular mechanisms, animal models are indispensable. Here, to identify genes silenced by promoter DNA methylation in rat mammary carcinomas, we took a combined approach of methylated DNA immunoprecipitation (MeDIP)-CpG island (CGI) microarray analysis and expression microarray analysis after treatment with epigenetic drugs. MeDIP-CGI microarray revealed that among 5031 genes with promoter CGI, 465 were methylated in a carcinoma cell line induced by 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), but not in normal mammary epithelial cells. By treatment of the cell line with 5-aza-2'-deoxycytidine and trichostatin A, 29 of the 465 genes were shown to be re-expressed. In primary mammary carcinomas, five (Angptl4, Coro1a, RGD1304982, Tmem37 and Ndn) of the 29 genes were methylated in one or more of 25 samples. Quantitative expression analysis revealed that Angptl4 had high expression in normal mammary glands, but low expression in primary carcinomas. Also in humans, ANGPTL4 was unmethylated and expressed in normal mammary epithelial cells, but was methylated in 11 of 91 (12%) primary breast cancers. This is the first study to identify genes aberrantly methylated in rat mammary carcinomas, and Angptl4 is a novel methylation-silenced gene both in rat and human mammary carcinomas. The combination of the MeDIP-CGI microarray analysis and expression microarray analysis after treatment with epigenetic drugs was effective in reducing the number of methylated genes that are not methylation silenced.


Lipolytic Markers, Insulin and Resting Fat Oxidation are Associated with Maximal Fat Oxidation.
The maximal capacity to oxidize fat during exercise (MFO) is associated with 24-h fat balance and insulin sensitivity. Understanding factors that influence MFO could have implications for metabolic health. We investigated relationships between selected plasma metabolites, hormones and overnight-fasted resting fat oxidation rates (Resting), with MFO. Resting fat oxidation and MFO was measured in 57 men with blood collected at rest and during exercise. Plasma glycerol (R=0.39, P=0.033), non-esterified fatty acids (NEFA: R=0.27, P=0.030) and insulin (R=- 0.36, P=0.007) measured at MFO correlated with MFO; only glycerol remained correlated when controlled for resting concentrations (R=0.36, P=0.008). The change in glycerol from rest to MFO correlated with exercise-induced fat oxidation (R=0.32, P=0.012). V˙O 2max correlated with resting fat oxidation (R=0.44, P=0.001) and MFO (R=0.52, P<0.001). Resting fat oxidation correlated with MFO (R=0.55, P<0.001); this remained when controlled for V˙O 2max (R=0.41, P=0.001). This study reports weak-to-moderate, albeit significant, relationships between plasma lipolytic markers, insulin and resting overnight-fasted fat oxidation with MFO and shows the plasma glycerol response to uniquely reflect exercise-induced fat oxidation. V˙O 2max correlates with fat oxidation but the relationship can be dissociated. Interventions to increase fat oxidation for optimal metabolic health would benefit from, but are not reliant on, increases in V˙O 2max.


Intraspecific variation in small-subunit rRNA sequences in GenBank: why single sequences may not adequately represent prokaryotic taxa.
Small-subunit rRNA (SSU rRNA) sequencing is a powerful tool to detect, identify, and classify prokaryotic organisms, and there is currently an explosion of SSU rRNA sequencing in the microbiology community. We report unexpectedly high levels of intraspecific variation (within and between strains) of prokaryote SSU rRNA sequences deposited in GenBank. A total of 82% of the prokaryote species with two published SSU rRNA sequences had more variable positions than a 0.1% random sequencing error would predict, and 48% of these sequence pairs had more variable positions than predicted by a 1.0% random sequencing error. Other sources of sequence variability must account for some of this intraspecific variation. Given these results, phylogenetic studies and biodiversity estimates obtained by using prokaryotic SSU rRNA sequences cannot proceed under the assumption that rRNA sequences of single operons from single isolates adequately represent their taxa. Sequencing SSU rRNA molecules from multiple operons and multiple isolates is highly recommended to obtain meaningful phylogenetic hypotheses, as is careful attention to accurate strain identification.


The oxygen uptake threshold during incremental exercise test.
The linear relationship between oxygen consumption (VO2) and exercise intensity is a well established phenomenon observed during incremental exercise. Recently, a non-linear increase in VO2 has been reported by Zoladz et al., who used a relatively complicated method to describe the phenomenon. In this study, we tried to ascertain whether the same phenomenon, which we named the oxygen uptake threshold (OUT), could be described by a simple method, using the two best fitting lines adopted for the less and more steep parts of the VO2 increase. Our hypothesis was that the non-linear VO2 increase was the result of a continuous VO2 increase (oxygen drift) occurring during the more intense steps only. Therefore, we analysed the VO2 time course during each step. Six cyclists performed an incremental exercise test on a cyclo-ergometer. The lactate threshold (LT) was calculated by using the intersection point of the two best fitting lines in the diagram of log LA (lactate concentration) dependence on log P (Power). The time course of VO2 during each step was analysed by an exponential rise to the maximum model. The results showed that OUT could be determined in five of the six subjects, whereas LT could be determined in all six subjects. The power output determined by OUT (168 +/- 13 W) was similar to that determined by LT (180 +/- 25 W). The VO2 time course during each step showed steady values during low intensity exercise. At intensities above LT and OUT, however, VO2 increased continuously, showing oxygen drift. It may be concluded that OUT is a realistic phenomenon, which is based on oxygen drift.


Activators of PKM2 in cancer metabolism.
Pyruvate kinase converts phosphoenolpyruvate to pyruvate, catalyzing the rate-limiting step of glycolysis. The M1 isoenzyme of pyruvate kinase (PKM1) is found in adult tissues; whereas, PKM2 is a splicesome variant found in embryonic and cancer cells. PKM2 expression in malignant cells is a result of the tumor microenvironment and is responsible for maintaining a glycolytic phenotype. PKM2 has other nonmetabolic functions in malignant cells, including transcriptional coactivation and protein kinase activity. PKM2 activators have antitumor properties by inducing tetramerization of two PKM2 dimers causing PKM2 to function like PKM1. Restoring PKM2 to PKM1-like levels of activity causes reversal of the Warburg effect in cancer cells. PKM2 activators have therapeutic potential in the treatment of cancer and other metabolic diseases.


Systemic Inflammatory Markers Predicting the Overall Survival of Patients Using Tyrosine Kinase Inhibitors in the First-line Treatment of Metastatic Renal Cell Carcinoma.
To identify prognostic inflammatory markers in metastatic renal cell carcinoma (mRCC) patients who received anti-vascular endothelial growth factor receptor (VEGFR) agents. Observational study. Place and Duration of the Study: Department of Medical Oncology, Necmettin Erbakan University, Meram Medical Faculty, Konya, Turkey, between January 2015 and December 2021. A total of 110 patients with mRCC who received sunitinib or pazopanib for at least 3 months were enrolled. Hemogram, C-reactive protein (CRP) and albumin values of the patients, CRP to albumin ratio (CAR), neutrophil-lymphocyte ratio (NLR), platelet-lymphocyte ratio (PLR), systemic immune-inflammation index (SII), prognostic nutrition index (PNI) and systemic inflammatory response indexes (SIRI) were calculated and recorded. Progression-free survival and overall survival analyses of the patients were performed using the Kaplan-Meier method. Cox regression method was used to identify prognostic factors. Variables found to be significant in univariate analysis were enrolled in multivariate analysis. In the univariate analysis for median overall survival (mOS), whether or not surgery was applied as the primary treatment option, grade, lymphovascular invasion (LVI), International Metastatic RCC Database Consortium (IMDC) score, CAR, NLR, PLR, SII, PNI and SIRI were found to be statistically significant. Systemic inflammation markers (CAR, NLR, PLR, PNI, SII and SIRI) were found to be independent prognostic markers for mOS as a result of Cox multivariate analysis. CAR, NLR, PLR, SII, PNI, and SIRI values measured before anti-VEGFR treatment in patients with mRCC may be of additional prognostic significance. These markers, which are calculated by using parameters that are always measured in routine practice, such as complete blood count (CBC), albumin, and CRP levels, are easy and inexpensive methods that give an idea about the course of the disease. Sunitinib, Pazopanib, Renal cell carcinoma, Prognostic marker, Overall survival, Inflammatory.


Exocyclically metallated tetrapyridinoporphyrazine as a potential photosensitizer for photodynamic therapy.
We report the first exocyclically metallated tetrapyridinoporphyrazine, [tetrakis-(trans-Pt(NH3)2Cl)-tetra(3,4-pyrido)porphyrazine-zinc(ii)](NO3)4 (4), synthesized in a multistep synthesis starting from 3,4-pyridinedicarbonitrile (1). The synthetic procedure involved a platination reaction of the intermediate tetra(3,4-pyrido)porphyrazine-zinc(ii) (2), whereby the zinc(ii) enhanced the solubility of the intermediate enabling the platination reaction. A similar approach to synthesize [tetrakis-(trans-Pt(NH3)2Cl)-tetra(3,4-pyrido)porphyrazine](NO3)4 (5) failed due to the unsuitable solubility properties of the intermediate tetra(3,4-pyrido)porphyrazine (3). The final product 4 and the intermediates were characterized, the photochemical and photophysical properties were determined and the photocytotoxicities were investigated. We demonstrate that the platinated tetra-pyridinoporphyrazine 4 is a potential photosensitizer for photodynamic therapy (PDT).


Plasma thrombomodulin concentrations in infants and children undergoing cardiac catheterization.
Circulating plasma thrombomodulin (TM) is an endothelial cell marker which may reflect endothelial injury. To find out to what extent diagnostic cardiac catheterization irritates vascular endothelium we conducted a prospective study in 91 children. Soluble TM concentrations, along with thrombin generation, were measured before, at the end of and 24 hours after cardiac catheterization. Compared to starting values, TM concentrations showed a clearly significant increase at the end of cardiac catheterization and returned to pretreatment values 24 hours later. Thrombin generation followed a similar pattern. Five out of the 91 children demonstrated resistance to activated protein C (APCR). With respect to the remaining 86 children, all five APCR cases showed increased thrombomodulin concentrations along with enhanced thrombin generation. Data from this study indicate that increased TM concentrations after cardiac catheterization in children are a sign of short-term endothelial damage. Furthermore, together with enhanced thrombin generation, elevated plasma concentration of soluble TM may reflect this receptor's possible anticoagulant properties.


Accuracy of autonomous robotic surgery for dental implant placement in fully edentulous patients: A retrospective case series study.
This study evaluated the accuracy of dental implant placement using the robotic computer-assisted implant surgery (r-CAIS) technology in fully edentulous patients. Fully edentulous and terminal dentition patients were enrolled for r-CAIS technology. Based on the cone-beam computed tomography (CBCT) examination, a customized positioning marker and a preoperative surgical plan were created before surgery. During the implant surgery, the implant osteotomy and placement were automatically performed using an autonomous robotic surgery system under the surgeon's supervision. A postoperative CBCT scan was used to determine the discrepancies between the planned and placed implants. Ten patients with 59 implants underwent autonomous robotic surgery. No adverse surgical events occurred. The deviations of global coronal, global apical, and angular were 0.67 ± 0.37 mm (95% CI: 0.58-0.77 mm), 0.69 ± 0.37 mm (95% CI: 0.59-0.78 mm), and 1.27° ± 0.59° (95% CI: 1.42°-1.11°), respectively. The autonomous r-CAIS technology proved an accurate surgical approach for implant placement in fully edentulous patients due to the control of the angular deviation. Autonomous robotic surgery seems promising as an accurate technology for treating fully edentulous patients. However, further trials are required to provide more hard clinical evidence.


Genome-wide association studies of mineral and phytic acid concentrations in pea (Pisum sativum L.) to evaluate biofortification potential.
Pea (Pisum sativum L.) is an important cool season food legume for sustainable food production and human nutrition due to its nitrogen fixation capabilities and nutrient-dense seed. However, minimal breeding research has been conducted to improve the nutritional quality of the seed for biofortification, and most genomic-assisted breeding studies utilize small populations with few single nucleotide polymorphisms (SNPs). Genomic resources for pea have lagged behind those of other grain crops, but the recent release of the Pea Single Plant Plus Collection (PSPPC) and the pea reference genome provide new tools to study nutritional traits for biofortification. Calcium, phosphorus, potassium, iron, zinc, and phytic acid concentrations were measured in a study population of 299 different accessions grown under greenhouse conditions. Broad phenotypic variation was detected for all parameters except phytic acid. Calcium exhibited moderate broad-sense heritability (H2) estimates, at 50%, while all other minerals exhibited low heritability. Of the accessions used, 267 were previously genotyped in the PSPPC release by the USDA, and we mapped the genotyping data to the pea reference genome for the first time. This study generated 54,344 high-quality SNPs used to investigate the population structure of the PSPPC and perform a genome-wide association study to identify genomic loci associated with mineral concentrations in mature pea seed. Overall, we were able to identify multiple significant SNPs and candidate genes for iron, phosphorus, and zinc. These results can be used for genetic improvement in pea for nutritional traits and biofortification, and the candidate genes provide insight into mineral metabolism.


Ischaemic nephropathy secondary to atherosclerotic renal artery stenosis: clinical and histopathological correlates.
Advanced renal artery stenosis (RAS) may cause progressive deterioration in renal function. We correlated the histopathological findings and clinical characteristics in selected patients with atherosclerotic RAS who underwent nephrectomy of their small kidneys for resistant renovascular hypertension. We studied 62 patients who underwent nephrectomy of a small kidney for uncontrolled hypertension between 1990 and 2000. The mean patient age was 65.4 ± 9.6 years; 28 (45%) were men. Significant tubulointerstitial atrophy with relative glomerular sparing was the predominant pattern of injury in 44 (71%) patients. In 14 (23%) patients, diffuse global glomerulosclerosis was present. The severity of tubulointerstitial atrophy and the extent of glomerulosclerosis were both associated with smaller kidney size (P = 0.002). Three patterns of vascular involvement were present: atheroembolic, atherosclerotic and hypertensive vascular changes, which were documented in 39, 98 and 52% of subjects, respectively. The presence and severity of these vascular changes positively correlated with both atherosclerotic risk factors, such as hypertension, dyslipidaemia and renal insufficiency, and cardiovascular morbidity, including abdominal aortic aneurysm and myocardial infarction. Patients on statin therapy were noted to have less evidence of renal fibrosis as measured by transforming growth factor-beta staining (P = 0.003). The severity of renal histopathological findings in patients who underwent nephrectomy for resistant hypertension correlated with an increased prevalence of cardiovascular disease, a greater degree of renal dysfunction and more severe dyslipidaemia. Statin therapy may affect development of intra-renal injury by slowing the progression of fibrosis.


Engineering dendritic cells to enhance cancer immunotherapy.
Cancer immunotherapy aims to establish immune-mediated control of tumor growth by priming T-cell responses to target tumor-associated antigens. Three signals are required for T-cell activation: (i) presentation of cognate antigen in self MHC molecules; (ii) costimulation by membrane-bound receptor-ligand pairs; and (iii) soluble factors to direct polarization of the ensuing immune response. The ability of dendritic cells (DCs) to provide all three signals required for T-cell activation makes them an ideal cancer vaccine platform. Several strategies have been developed to enhance and control antigen presentation, costimulation, and cytokine production. In this review, we discuss progress toward developing DC-based cancer vaccines by genetic modification using RNA, DNA, and recombinant viruses. Furthermore, the ability of DC-based vaccines to activate natural killer (NK) and B-cells, and the impact of gene modification strategies on these populations is described. Clinical trials using gene-modified DCs have shown modest results, therefore, further considerations for DC manipulation to enhance their clinical efficacy are also discussed.


Characterization of two Campylobacter jejuni strains for use in volunteer experimental-infection studies.
The development of vaccines against Campylobacter jejuni would be facilitated by the ability to perform phase II challenge studies. However, molecular mimicry of the lipooligosaccharide (LOS) of most C. jejuni strains with human gangliosides presents safety concerns about the development of Guillain-Barré syndrome. Clinical isolates of C. jejuni that appeared to lack genes for the synthesis of ganglioside mimics were identified by DNA probe analyses. Two clinical isolates from Southeast Asia (strains BH-01-0142 and CG8421) were determined to express the LOS type containing N-acetyl quinovosamine. No ganglioside structures were observed to be present in the LOSs of these strains, and pyrosequence analyses of the genomes of both strains confirmed the absence of genes involved in ganglioside mimicry. The capsule polysaccharide (CPS) of BH-01-0142 was determined to be composed of galactose (Gal), 6-deoxy-ido-heptose, and, in smaller amounts, D-glycero-D-ido-heptose, and the CPS of CG8421 was observed to contain Gal, 6-deoxy-altro-heptose, N-acetyl-glucosamine, and minor amounts of 6-deoxy-3-O-Me-altro-heptose. Both CPSs were shown to carry O-methyl-phosphoramidate. The two genomes contained strain-specific zones, some of which could be traced to a plasmid origin, and both contained a large chromosomal insertion related to the CJEI3 element of C. jejuni RM1221. The genomes of both strains shared a high degree of similarity to each other and, with the exception of the capsule locus of CG8421, to the type strain of the HS3 serotype, TGH9011.


How, in what contexts, and why do quality dashboards lead to improvements in care quality in acute hospitals? Protocol for a realist feasibility evaluation.
National audits are used to monitor care quality and safety and are anticipated to reduce unexplained variations in quality by stimulating quality improvement (QI). However, variation within and between providers in the extent of engagement with national audits means that the potential for national audit data to inform QI is not being realised. This study will undertake a feasibility evaluation of QualDash, a quality dashboard designed to support clinical teams and managers to explore data from two national audits, the Myocardial Ischaemia National Audit Project (MINAP) and the Paediatric Intensive Care Audit Network (PICANet). Realist evaluation, which involves building, testing and refining theories of how an intervention works, provides an overall framework for this feasibility study. Realist hypotheses that describe how, in what contexts, and why QualDash is expected to provide benefit will be tested across five hospitals. A controlled interrupted time series analysis, using key MINAP and PICANet measures, will provide preliminary evidence of the impact of QualDash, while ethnographic observations and interviews over 12 months will provide initial insight into contexts and mechanisms that lead to those impacts. Feasibility outcomes include the extent to which MINAP and PICANet data are used, data completeness in the audits, and the extent to which participants perceive QualDash to be useful and express the intention to continue using it after the study period. The study has been approved by the University of Leeds School of Healthcare Research Ethics Committee. Study results will provide an initial understanding of how, in what contexts, and why quality dashboards lead to improvements in care quality. These will be disseminated to academic audiences, study participants, hospital IT departments and national audits. If the results show a trial is feasible, we will disseminate the QualDash software through a stepped wedge cluster randomised trial.


Epidemiology and outcomes of community-acquired Clostridium difficile infections in Medicare beneficiaries.
The incidence of community-acquired Clostridium difficile (CACD) is increasing in the United States. Many CACD infections occur in the elderly, who are predisposed to poor outcomes. We aimed to describe the epidemiology and outcomes of CACD in a nationally representative sample of Medicare beneficiaries. We queried a 5% random sample of Medicare beneficiaries (2009-2011 Part A inpatient and Part D prescription drug claims; n = 864,604) for any hospital admission with a primary ICD-9 diagnosis code for C difficile (008.45). We examined patient sociodemographic and clinical characteristics, preadmission exposure to oral antibiotics, earlier treatment with oral vancomycin or metronidazole, inpatient outcomes (eg, colectomy, ICU stay, length of stay, mortality), and subsequent admissions for C difficile. A total of 1,566 (0.18%) patients were admitted with CACD. Of these, 889 (56.8%) received oral antibiotics within 90 days of admission. Few were being treated with oral metronidazole (n = 123 [7.8%]) or vancomycin (n = 13 [0.8%]) at the time of admission. Although 223 (14%) patients required ICU admission, few (n = 15 [1%]) underwent colectomy. Hospital mortality was 9%. Median length of stay among survivors was 5 days (interquartile range 3 to 8 days). One fifth of survivors were readmitted with C difficile, with a median follow-up time of 393 days (interquartile range 129 to 769 days). Nearly half of the Medicare beneficiaries admitted with CACD have no recent antibiotic exposure. High mortality and readmission rates suggest that the burden of C difficile on patients and the health care system will increase as the US population ages. Additional efforts at primary prevention and eradication might be warranted.


[Treatment Refusal in Pediatric Oncology].
Pediatric oncologists are often faced with situations in which parents or guardians refuse recommended treatment for curable childhood cancer. Deciding how to proceed in such situations is an ethical dilemma. The aim of this article is to consider optimal approaches when parents are strongly against oncological treatment, potentially compromising their childrens rights for health care and to the chance for cure. In this paper, we report two cases of treatment refusal from our department and the impact of such decisions on the children themselves. Case no. 1 describes a child with retinoblastoma whose parents refused standard treatment in order to seek alternative treatment abroad. Case no. 2 describes a patient with a primary lymphoma of bone who received treatment by a court order after parental refusal. When parents refuse a treatment for potentially curable cancer, the medical team often focuses on the certainty of death without treatment. In the background, there is a smaller but still significant risk that - even if the treatment is eventually accepted or compelled - the child will still die from treatment-related complications or refractory disease, possibly with considerable suffering. The reasons for refusing a treatment vary. The entire medical team is tasked with trying to respectfully understand the reasoning behind the parents unwillingness to accept the treatment, in order to address all possible misunderstandings and to propose solutions that could be acceptable for the parents. In some situations however, it is necessary to resolve the dilemma by legal means in order to protect the life of the child.Key words: oncology - ethics - decision making - treatment refusal - legal guardians The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study. The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers.Submitted: 7. 8. 2017Accepted: 7. 9. 2017.


Determination of antithrombin-dependent factor Xa inhibitors by prothrombin-induced clotting time.
Prothrombinase-induced clotting time (PiCT) determines the anticoagulant effects of heparins, low molecular weight heparins (LMWHs), and direct thrombin inhibitors. At present, this is the only method that measures the effects of all of these inhibitors, in contrast to the prothrombin time, activated partial thromboplastin time (aPTT), Heptest, ecarin clotting time, and the chromogenic assays. The antithrombin-dependent direct factor (F) Xa inhibitors fondaparinux and idraparinux were compared with the LMWH dalteparin on PiCT, aPTT, Heptest, and chromogenic anti-FXa assays in pooled human normal plasma samples. Fondaparinux and idraparinux prolonged the coagulation times in the PiCT, Heptest, and chromogenic FXa assays in a dose-dependent manner, in contrast to the aPTT. We conclude that PiCT is a suitable assay to determine the anticoagulant effects of these two new FXa inhibitors in patients receiving treatment with these compounds.


Topical dermal application of essential oils attenuates the severity of adjuvant arthritis in Lewis rats.
This study was aimed at examining the effect of an ointment containing essential oils (EO) on the severity of adjuvant arthritis (AA), an experimental model of human rheumatoid arthritis (RA), in Lewis rats and to define the underlying mechanisms. At the onset of AA, the rats received topical application twice daily of an ointment containing 20% EO or placebo ointment. The synovial fluid (SF) and synovium-infiltrating cells (SIC) of rats were tested for pro-inflammatory cytokines TNF-α and IL-1β. The hind paws and skin were examined histologically. The activity/level of matrix metalloproteinases (MMPs) and anti-mycobacterial heat-shock protein 65 (Bhsp65) antibodies were tested. Arthritic rats treated with ointment containing EO developed less severe clinical arthritis compared with the controls, and this activity was attributable to EO and not to the carrier oil. The levels of TNF-α and IL-1β, and the activity of MMPs in SF and SIC-lysate were significantly reduced in EO-treated arthritic rats compared with the controls. However, the levels of anti-Bhsp65 antibodies were unaffected by treatment. Thus, topical dermal delivery of EO-containing ointment down-modulates the severity of AA in Lewis rats by inhibiting defined mediators of inflammation. Such ointments should be tested in patients with RA and other arthritic conditions.


The application of two-step linear temperature program to thermal analysis for monitoring the lipid induction of Nostoc sp. KNUA003 in large scale cultivation.
Recently, microalgae was considered as a renewable energy for fuel production because its production is nonseasonal and may take place on nonarable land. Despite all of these advantages, microalgal oil production is significantly affected by environmental factors. Furthermore, the large variability remains an important problem in measurement of algae productivity and compositional analysis, especially, the total lipid content. Thus, there is considerable interest in accurate determination of total lipid content during the biotechnological process. For these reason, various high-throughput technologies were suggested for accurate measurement of total lipids contained in the microorganisms, especially oleaginous microalgae. In addition, more advanced technologies were employed to quantify the total lipids of the microalgae without a pretreatment. However, these methods are difficult to measure total lipid content in wet form microalgae obtained from large-scale production. In present study, the thermal analysis performed with two-step linear temeperature program was applied to measure heat evolved in temperature range from 310 to 351 °C of Nostoc sp. KNUA003 obtained from large-scale cultivation. And then, we examined the relationship between the heat evolved in 310-351 °C (HE) and total lipid content of the wet Nostoc cell cultivated in raceway. As a result, the linear relationship was determined between HE value and total lipid content of Nostoc sp. KNUA003. Particularly, there was a linear relationship of 98% between the HE value and the total lipid content of the tested microorganism. Based on this relationship, the total lipid content converted from the heat evolved of wet Nostoc sp. KNUA003 could be used for monitoring its lipid induction in large-scale cultivation.


Helping patients simplify and safely use complex prescription regimens.
There is considerable variability in the manner in which prescriptions are written by physicians and transcribed by pharmacists, resulting in patient misunderstanding of label instructions. A universal medication schedule was recently proposed for standardizing prescribing practices to 4 daily time intervals, thereby helping patients simplify and safely use complex prescription regimens. We investigated whether patients consolidate their medications or whether there is evidence of unnecessary regimen complexity that would support standardization. Structured interviews were conducted with 464 adults (age range, 55-74 years) who were receiving care either at an academic general medicine practice or at 1 of 3 federally qualified health centers in Chicago, Illinois. Participants were given a hypothetical, 7-drug medication regimen and asked to demonstrate how and when they would take all of the medications in a 24-hour period. The regimen could be consolidated into 4 dosing episodes per day. The primary outcome was the number of times per day that individuals would take medication. Root causes for patients complicating the regimen (>4 times a day) were examined. Participants on average identified 6 times (SD, 1.8 times; range, 3-14 times) in 24 hours to take the 7 drugs. One-third of the participants (29.3%) dosed their medications 7 or more times per day, while only 14.9% organized the regimen into 4 or fewer times a day. In multivariable analysis, low literacy was an independent predictor of more times per day for dosing the regimen (β = 0.67; 95% confidence interval, 0.12-1.22; P = .02). Instructions for 2 of the drugs were identical, yet 31.0% of the participants did not take these medications at the same time. Another set of drugs had similar instructions, with the primary exception of 1 drug having the added instruction to take "with food and water." Half of the participants (49.5%) took these medications at different times. When the medications had variable expressions of the same dose frequency (eg, "every 12 hours" vs "twice daily"), 79.0% of the participants did not consolidate the medications. Many patients, especially those with limited literacy, do not consolidate prescription regimens in the most efficient manner, which could impede adherence. Standardized instructions proposed with the universal medication schedule and other task-centered strategies could potentially help patients routinely organize and take medication regimens.


Essential role of IFN-γ in T cell-associated intestinal inflammation.
Paneth cells contribute to small intestinal homeostasis by secreting antimicrobial peptides and constituting the intestinal stem cell (ISC) niche. Certain T cell-mediated enteropathies are characterized by extensive Paneth cell depletion coincident with mucosal destruction and dysbiosis. In this study, mechanisms of intestinal crypt injury have been investigated by characterizing responses of mouse intestinal organoids (enteroids) in coculture with mouse T lymphocytes. Activated T cells induced enteroid damage, reduced Paneth cell and Lgr5+ ISC mRNA levels, and induced Paneth cell death through a caspase-3/7-dependent mechanism. IFN-γ mediated these effects, because IFN-γ receptor-null enteroids were unaffected by activated T cells. In mice, administration of IFN-γ induced enteropathy with crypt hyperplasia, villus shortening, Paneth cell depletion, and modified ISC marker expression. IFN-γ exacerbated radiation enteritis, which was ameliorated by treatment with a selective JAK1/2 inhibitor. Thus, IFN-γ induced Paneth cell death and impaired regeneration of small intestinal epithelium in vivo, suggesting that IFN-γ may be a useful target for treating defective mucosal regeneration in enteric inflammation.


Inactivation of poxviruses by upper-room UVC light in a simulated hospital room environment.
In the event of a smallpox outbreak due to bioterrorism, delays in vaccination programs may lead to significant secondary transmission. In the early phases of such an outbreak, transmission of smallpox will take place especially in locations where infected persons may congregate, such as hospital emergency rooms. Air disinfection using upper-room 254 nm (UVC) light can lower the airborne concentrations of infective viruses in the lower part of the room, and thereby control the spread of airborne infections among room occupants without exposing occupants to a significant amount of UVC. Using vaccinia virus aerosols as a surrogate for smallpox we report on the effectiveness of air disinfection, via upper-room UVC light, under simulated real world conditions including the effects of convection, mechanical mixing, temperature and relative humidity. In decay experiments, upper-room UVC fixtures used with mixing by a conventional ceiling fan produced decreases in airborne virus concentrations that would require additional ventilation of more than 87 air changes per hour. Under steady state conditions the effective air changes per hour associated with upper-room UVC ranged from 18 to 1000. The surprisingly high end of the observed range resulted from the extreme susceptibility of vaccinia virus to UVC at low relative humidity and use of 4 UVC fixtures in a small room with efficient air mixing. Increasing the number of UVC fixtures or mechanical ventilation rates resulted in greater fractional reduction in virus aerosol and UVC effectiveness was higher in winter compared to summer for each scenario tested. These data demonstrate that upper-room UVC has the potential to greatly reduce exposure to susceptible viral aerosols. The greater survival at baseline and greater UVC susceptibility of vaccinia under winter conditions suggest that while risk from an aerosol attack with smallpox would be greatest in winter, protective measures using UVC may also be most efficient at this time. These data may also be relevant to influenza, which also has improved aerosol survival at low RH and somewhat similar sensitivity to UVC.


Fuzzy, copper-based multi-functional composite particles serving simultaneous catalytic and signal-enhancing roles.
Multifunctional plasmonic particles serving simultaneously as catalysts and label-free reporting agents are highly pursued due to their great potential in enhancing reaction operational efficiencies. Copper is an abundant and economic resource, and it possesses practical applicability in industries, but no dual-functional copper-based catalytic and self-reporting particles have been reported so far. This study proposes a facile strategy to prepare high-performance dual-functional copper-based composite particles that catalyze reactions and simultaneously serve as a SERS (surface enhanced Raman spectra) active, label-free reporting agent. Polyelectrolyte-modified reduced graphene oxide particles are used as the reactive precursors in the fabrication method. Upon adding Cu(NO3)2 solutions into the precursor dispersions, composite particles comprised by copper/copper oxide core and polyelectrolyte-graphene shell were facilely obtained under sonication. The as-prepared composite particles efficiently catalyzed the conversion of 4-nitrophenol to 4-aminophenol and simultaneously acted as the SERS-active substrate to give enhanced Raman spectra of the produced 4-aminophenol. Taking advantage of the assembling capabilities of polyelectrolyte shells, the composite particles could be further assembled onto a planar substrate to catalyze organic reactions, facilitating their application in various conditions. We expect this report to promote the fabrication and application of copper-based multifunctional particles.


Dalfampridine improves walking speed, walking endurance, and community participation in veterans with multiple sclerosis: a longitudinal cohort study.
In short-term trials, dalfampridine extended release (ER) improves walking in people with multiple sclerosis (MS). The tolerability and effects of dalfampridine-ER in clinical practice have not been reported. The objective of this paper is to determine the clinical tolerability and effects of dalfampridine on walking and community participation. All patients at the Portland VA Medical Center prescribed dalfampridine-ER over one year completed the Timed 25-Foot Walk (T25FW), Multiple Sclerosis Walking Scale-12 (MSWS-12), Two-Minute Timed Walk (2MTW), and Community Integration Questionnaire (CIQ) at baseline and follow-up clinic visits. Ongoing use and measures over one year were analyzed. A total of 39 patients (mean age 56.5 years, mean disease duration 19.5 years, 82% male, 38% relapsing-remitting MS, 62% progressive MS) were prescribed dalfampridine-ER. Twenty-four (62%) continued to take dalfampridine-ER. At initial follow-up, all measures improved significantly from baseline (T25FW: -2.7 s, p = 0.004; 2MTW: 41 feet (ft), p = 0.002; MSWS12: -11, p < 0.001; CIQ: 1.2, p = 0.003). At one year, walking endurance and self-perceived walking were still significantly improved (2MTW: 33 ft, p = 0.03; MSWS-12: 5.9, p = 0.007). Dalfampridine-ER was associated with short-term improvements in walking speed and community participation, and sustained improvements in walking endurance and self-perceived impact of MS on walking for one year. Our study supports the utility of this medication in late MS.


Physician code creep after the initiation of outpatient volume control program and implications for appropriate ICD-10-CM coding.
Most studies on the physician code creep (i.e., changes in case mix record-keeping practices to improve reimbursement) have focused on episodes (inpatient hospitalizations or outpatient procedures). Little is known regarding changes in diagnostic coding practices for better reimbursement among a fixed cohort of patients with chronic diseases. To examine whether physicians in tertiary medical centers changed their coding practices after the initiation of the Outpatient Volume Control Program (OVCP) in Taiwan, we conducted a retrospective observational study of four patient cohorts (two interventions and two controls) from January 2016 to September 2017 in Taiwan. The main outcomes were the number of outpatient visits with four coding practices: 1) OVCP monitoring code recorded as primary diagnosis; 2) OVCP monitoring code recorded as secondary diagnosis; 3) non-OVCP monitoring code recorded as primary diagnosis; 4) non-OVCP monitoring code recorded as secondary diagnosis. The percentage change of the number of visits with coding practice 1 between 2016Q1 and 2017Q3 was - 74% for patients with hypertension and - 73% with diabetes in tertiary medical centers and - 23% and - 17% in clinics, respectively. By contrast, the percentage changes of coding practice 3 were + 73% for patients with hypertension and + 46% for patients with diabetes in tertiary medical centers and - 19% and - 2% in clinics, respectively. Physician code creep occurred after the initiation of the OVCP. Education regarding appropriate outpatient coding for physicians will be relatively effective when proper coding is related to reimbursement.


Duration of illness and structure of symptoms in schizophrenia.
Previous research has mainly focused on the cross-sectional structure of symptoms in schizophrenia. This meta-analysis examined the association of duration of illness with the structure of symptoms. Using explicit criteria, 22 studies reporting on the correlations of symptoms in 2665 schizophrenic patients were selected. From each study, symptom-pair correlations for negative symptoms as rated by Scale for the Assessment of Negative Symptoms (SANS) and positive symptoms as rated by the Scale for the Assessment of Positive Symptoms (SAPS) were extracted. Variability among symptom-pair correlations across studies was assessed using tests of homogeneity. For symptom-pair correlations which were not found to be homogeneous, the association of average duration of illness with the symptom-pair correlations were examined. There was considerable variability in symptom-pair correlations across studies. Part of this variability was explainable by variations in average duration of illness. Longer duration of illness was associated with lower negative-negative symptom-pair correlations and higher negative-positive symptom-pair correlations. The findings suggest that the structure of symptoms in schizophrenia evolves over time, following a consistent pattern. In the early stages of illness, negative and positive symptoms form cohesive dimensions. With time, these dimensions become less cohesive and the boundaries between them, less clear.


Immune responses to Plasmodium vivax pre-erythrocytic stage antigens in naturally exposed Duffy-negative humans: a potential model for identification of liver-stage antigens.
Duffy antigen is the receptor used by Plasmodium vivax to invade erythrocytes. Consequently, individuals lacking Duffy antigen [Fy(-)] do not develop blood-stage infections. We hypothesized that naturally exposed Fy(-) humans may develop immune responses mainly to pre-erythrocytic stages and could be used to study acquired immunity to P. vivax and to identify liver-stage antigens. We report here that antibody and IFN-gamma responses to known sporozoite antigens were significantly induced by natural exposure in Fy(-) humans, whereas responses to blood-stage antigens were significantly induced in Fy(+) humans. IFN-gamma responses to sporozoite antigens were lower in Fy(+) than in Fy(-) humans, indicating that in Fy(+) humans blood-stage infections may have suppressed T cell responses to pre-erythrocytic stages. We evaluated the immune responses to 18 novel P. vivax homologs of P. falciparum sporozoite proteins identified from the P. vivax genome sequence. Eight proteins recalled IFN-gamma responses in P. vivax-exposed but not in unexposed individuals. Of these, 3 antigens elicited IFN-gamma responses in Fy(-) but not in Fy(+) individuals. These results suggest that differential immune responses observed in naturally exposed Fy(-) and Fy(+) individuals can be exploited to identify P. vivax stage-specific antigens.


Evolutionary diversification of type 2 porcine reproductive and respiratory syndrome virus.
Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the leading swine pathogens causing tremendous economic loss to the global swine industry due to its virulence, pathogenesis, infectivity and transmissibility. Although formally recognized only two and half decades ago, molecular dating estimation indicates a more ancient evolutionary history, which involved divergence into two genotypes (type 1 and type 2) prior to the 'initial' outbreaks of the late 1980s. Type 2 PRRSV circulates primarily in North America and Asia. The relatively greater availability of sequence data for this genotype from widespread geographical territories has enabled a better understanding of the evolving genotype. However, there are a number of challenges in terms of the vastness of data available and what this indicates in the context of viral diversity. Accordingly, here we revisit the mechanisms by which PRRSV generates variability, describe a means of organizing type 2 diversity captured in voluminous ORF5 sequences in a phylogenetic framework and provide a holistic view of known global type 2 diversity in the same setting. The consequences of the expanding diversity for control measures such as vaccination are discussed, as well as the contribution of modified live vaccines to the circulation of field isolates. We end by highlighting some limitations of current molecular epidemiology studies in relation to inferring PRRSV diversity, and what steps can be taken to overcome these and additionally enable PRRSV sequence data to be informative about viral phenotypic traits such as virulence.


[Thrombocytic and coagulation hemostasis in pregnant women living in the Ob-Irtysh river basin with chronic opisthorchiasis].
Opisthorchiasis that is widely spreading in Russia and CIS countries was and remains a serious social, medical, and scientific problem. Local or imported cases of opisthorchiasis have been notified in 87.6% of the administrative territories of Russia. The world's largest focus of opisthorchiasis is the Ob-Irtysh river basin with natural preconditions for its high infection in fish. The main clinical manifestations of Opisthorchis infestation are associated with liver damage caused by the obligate presence of Opisthorchis in the bile capillaries with evolving cholangitis, cholangiocholecystitis, and cholangiopancreatitis. Opisthorchis complicates the course of many diseases, affecting gestation, labor, and postpartum. The state of thrombocytic and coagulation hemostasis was analyzed in different period of pregnancy and postpartum (days 2-4), by using the course of pregnancy in the inhabitants of the hyperendemic focus of Opisthorchis infestation (the Ob-Irtysh basin) as an example. In the newcomers, pregnancy develops in the presence of platelet hyperaggregation and accelerated continuous blood clotting. In pregnant women who belong to native little peoples (Khanty, Mansi), opisthorchiasis induces no significant changes in the hemostatic potential.


Regulation of GATA-3 expression during CD4 lineage differentiation.
GATA-3 is necessary for the development of MHC class II-restricted CD4 T cells, and its expression is increased during positive selection of these cells. TCR signals drive this upregulation, but the signaling pathways that control this process are not well understood. Using genetic and pharmacological approaches, we show that GATA-3 upregulation during thymocyte-positive selection is the result of additive inputs from the Ras/MAPK and calcineurin pathways. This upregulation requires the presence of the transcription factor c-Myb. Furthermore, we show that TH-POK can also upregulate GATA-3 in double-positive thymocytes, suggesting the existence of a positive feedback loop that contributes to lock in the initial commitment to the CD4 lineage during differentiation.


Physiological and transcriptomic characterization of a fliA mutant of Pseudomonas putida KT2440.
Pseudomonas putida KT2440 encodes 23 alternative sigma factors. The fliA gene, which encodes σ(28) , is in a cluster with other genes involved in flagella biosynthesis and chemotaxis. Reverse transcriptase-PCR revealed that this cluster is comprised of four independent transcriptional units: flhAF, fleNfliA, cheYZA and cheBmotAB. We generated a nonpolar fliA mutant by homologous recombination and tested its motility, adhesion to biotic and abiotic surfaces, and responses to various stress conditions. The mutant strain was nonmotile and exhibited decreased capacity to bind to corn seeds, although its ability to colonize the rhizosphere of plants was unaffected. The mutant was also affected in binding to abiotic surfaces and its ability to form biofilms decreased by almost threefold. In the fliA mutant background expression of 25 genes was affected: two genes were upregulated and 23 genes were downregulated. In addition to a number of motility and chemotaxis genes, the fliA gene product is also necessary for the expression of some genes potentially involved in amino acid utilization or stress responses; however, we were unable to assign specific phenotypes linked to these genes since the fliA mutant used the same range of amino acids as the parental strain, and was as tolerant as the wild type to stress imposed by heat, antibiotics, NaCl, sodium dodecyl sulfate, H2 O2 and benzoate. Based on the sequence alignment of promoters recognized by FliA and genome in silico analysis, we propose that P. putidaσ(28) recognizes a TCAAG-t-N12 -GCCGATA consensus sequence located between -34 and -8 and that this sequence is preferentially associated with an AT-rich upstream region.


Loss of hepatic Mboat7 leads to liver fibrosis.
The rs641738C>T variant located near the membrane-bound O-acyltransferase domain containing 7 (MBOAT7) locus is associated with fibrosis in liver diseases, including non-alcoholic fatty liver disease (NAFLD), alcohol-related liver disease, hepatitis B and C. We aim to understand the mechanism by which the rs641738C>T variant contributes to pathogenesis of NAFLD. Mice with hepatocyte-specific deletion of MBOAT7 (Mboat7Δhep) were generated and livers were characterised by histology, flow cytometry, qPCR, RNA sequencing and lipidomics. We analysed the association of rs641738C>T genotype with liver inflammation and fibrosis in 846 NAFLD patients and obtained genotype-specific liver lipidomes from 280 human biopsies. Allelic imbalance analysis of heterozygous human liver samples pointed to lower expression of the MBOAT7 transcript on the rs641738C>T haplotype. Mboat7Δhep mice showed spontaneous steatosis characterised by increased hepatic cholesterol ester content after 10 weeks. After 6 weeks on a high fat, methionine-low, choline-deficient diet, mice developed increased hepatic fibrosis as measured by picrosirius staining (p<0.05), hydroxyproline content (p<0.05) and transcriptomics, while the inflammatory cell populations and inflammatory mediators were minimally affected. In a human biopsied NAFLD cohort, MBOAT7 rs641738C>T was associated with fibrosis (p=0.004) independent of the presence of histological inflammation. Liver lipidomes of Mboat7Δhep mice and human rs641738TT carriers with fibrosis showed increased total lysophosphatidylinositol levels. The altered lysophosphatidylinositol and phosphatidylinositol subspecies in MBOAT7Δhep livers and human rs641738TT carriers were similar. Mboat7 deficiency in mice and human points to an inflammation-independent pathway of liver fibrosis that may be mediated by lipid signalling and a potentially targetable treatment option in NAFLD.


Nitrate losses in subsurface drainage from a corn-soybean rotation as affected by fall and spring application of nitrogen and nitrapyrin.
Substantial amounts of NO3 from agricultural crop production systems on poorly drained soils can be transported to surface water via subsurface drainage. A field study was conducted from the fall of 1993 through 2000 on a tile-drained Canisteo clay loam soil (fine-loamy, mixed, superactive, calcareous, mesic Typic Endoaquoll) to determine the influence of fall vs. spring application of N and nitrapyrin [NP; 2-chloro-6-(trichloromethyl) pyridine] on NO3 losses from a corn (Zea mays L.)-soybean [Glycine max (L.) Merr.] rotation. Four anhydrous ammonia treatments (fall N, fall N + NP, spring preplant N, and spring N + NP) were replicated four times and applied at 135 kg N ha(-1) for corn on individual drainage plots. Drainage occurred in all seven years. Seventy-one percent of the annual drainage and 75% of the annual NO3 loss occurred in April, May, and June. Fifty-four percent of the NO3 lost in the drainage occurred during the corn phase and 46% during the soybean phase. Annual flow-weighted NO3-N concentrations for the fall, fall + NP, spring, and spring + NP treatments averaged 14.3, 11.5, 10.7, and 11.3 mg L(-1) during the corn phase but annual NO3-N concentrations were still > or =10 mg L(-1) in three of six years for the spring preplant treatment. Averaged across the six rotation cycles, flow-normalized NO3-N losses ranked in the order: fall N > spring N + NP > fall N + NP > spring N. Under these conditions, NO3 losses in subsurface drainage from a corn-soybean rotation can be reduced 14% by spring N and 10% by late fall N + NP compared with fall-applied N. Nitrate losses were not appreciably reduced by adding NP to spring preplant N.


Exploration of stilbenoid trimers as potential inhibitors of sirtuin1 enzyme using a molecular docking and molecular dynamics simulation approach.
A combination of molecular docking and molecular dynamics simulation (250 ns) has been carried out to study the interaction of stilbenoid trimer compounds with the SIRT1 enzyme as the target protein. SIRT1 expression regulates cellular stress responses that lead to the development of cancer. Redocking showed a good native ligand pose with an RMSD value of 1.40 Å at the receptor active site's coordinates. The molecular docking score uses a grid score functional (kcal mol-1), which shows results of 1NS: 79.56, TS1: -26.83, TS2: -87.77, and TS3: -83.67. The TS2 and TS3 candidates were chosen for further analysis because they had a lower grid score than the native ligand (1NS). Furthermore, prediction of binding free energy (kcal mol-1) using the Quantum Mechanics/generalized Born Surface Area (QM/MM-GBSA) method shows the results of 1NS: -31.52 ± 0.39, TS2: -58.99 ± 0.34, and TS3: -43.38 ± 0.35. These results indicate that the TS2 and TS3 compounds have good potential as inhibitors of the SIRT1 enzyme. Additionally, the amino acid residues were responsible for the inhibition mechanism through hydrogen bond interactions at the molecular level, including ASP22, PHE91, PRO11, ILE165, ASP166, and VAL230. The observations made in this study provide theoretical information for exploring the stilbenoid trimers as anticancer agents by targeting the SIRT1 enzyme.


Further characterization of tissue distribution and metabolism of (14C)aflatoxin B1 in chickens.
The distribution and metabolism of [(14)C]aflatoxin B(1) in chicken tissues were further investigated. Previously dried and frozen ethyl acetate extracts of liver, heart, gizzard, breast, leg, blood, and fecal samples were obtained from either layer or broiler chickens fed subclinical levels of [(14)C]aflatoxin B(1). Treatment of these extracts with either carboxypeptidase A, leucine aminopeptidase, pepsin, or trypsin revealed that an average of 50% of the (14)C detected in the acetate extracts was a liberated peptide (or amino acid) conjugate of [(14)C]aflatoxin B(2a). When a prepared standard of B(2a) was made by incubation of B(1) with cold dilute aqueous HCl, the R(f) values and absorbance maxima were identical with those of the tissue extracts after enzymatic treatment.


Kytococcus schroeteri prosthetic valve endocarditis.
We report the case of a 64-year-old male patient with a prosthetic aortic valve who presented with clinical features of endocarditis confirmed by transoesophageal echocardiography. His blood cultures were positive for a very rare and newly described organism-Kytococcus schroeteri. The patient underwent aortic valve replacement and a 6-week course of intravenous antibiotics. This is the fifth reported case of endocarditis associated with this organism.