Prehistoric mitochondrial DNA of domesticate animals supports a 13th century exodus from the northern US southwest.
The 13th century Puebloan depopulation of the Four Corners region of the US Southwest is an iconic episode in world prehistory. Studies of its causes, as well as its consequences, have a bearing not only on archaeological method and theory, but also social responses to climate change, the sociology of social movements, and contemporary patterns of cultural diversity. Previous research has debated the demographic scale, destinations, and impacts of Four Corners migrants. Much of this uncertainty stems from the substantial differences in material culture between the Four Corners vs. hypothesized destination areas. Comparable biological evidence has been difficult to obtain due to the complete departure of farmers from the Four Corners in the 13th century CE and restrictions on sampling human remains. As an alternative, patterns of genetic variation among domesticated species were used to address the role of migration in this collapse. We collected mitochondrial haplotypic data from dog (Canis lupus familiaris) and turkey (Meleagris gallopavo) remains from archaeological sites in the most densely-populated portion of the Four Corners region, and the most commonly proposed destination area for that population under migration scenarios. Results are consistent with a large-scale migration of humans, accompanied by their domestic turkeys, during the 13th century CE. These results support scenarios that suggest contemporary Pueblo peoples of the Northern Rio Grande are biological and cultural descendants of Four Corners populations.


Analysis of phosphatases in ER-negative breast cancers identifies DUSP4 as a critical regulator of growth and invasion.
Estrogen receptor (ER)-negative cancers have a poor prognosis, and few targeted therapies are available for their treatment. Our previous analyses have identified potential kinase targets critical for the growth of ER-negative, progesterone receptor (PR)-negative and HER2-negative, or "triple-negative" breast cancer (TNBC). Because phosphatases regulate the function of kinase signaling pathways, in this study, we investigated whether phosphatases are also differentially expressed in ER-negative compared to those in ER-positive breast cancers. We compared RNA expression in 98 human breast cancers (56 ER-positive and 42 ER-negative) to identify phosphatases differentially expressed in ER-negative compared to those in ER-positive breast cancers. We then examined the effects of one selected phosphatase, dual specificity phosphatase 4 (DUSP4), on proliferation, cell growth, migration and invasion, and on signaling pathways using protein microarray analyses of 172 proteins, including phosphoproteins. We identified 48 phosphatase genes are significantly differentially expressed in ER-negative compared to those in ER-positive breast tumors. We discovered that 31 phosphatases were more highly expressed, while 11 were underexpressed specifically in ER-negative breast cancers. The DUSP4 gene is underexpressed in ER-negative breast cancer and is deleted in approximately 50 % of breast cancers. Induced DUSP4 expression suppresses both in vitro and in vivo growths of breast cancer cells. Our studies show that induced DUSP4 expression blocks the cell cycle at the G1/S checkpoint; inhibits ERK1/2, p38, JNK1, RB, and NFkB p65 phosphorylation; and inhibits invasiveness of TNBC cells. These results suggest that that DUSP4 is a critical regulator of the growth and invasion of triple-negative breast cancer cells.


Mathematical modeling of circadian rhythms.
Circadian rhythms are endogenous ~24-hr oscillations usually entrained to daily environmental cycles of light/dark. Many biological processes and physiological functions including mammalian body temperature, the cell cycle, sleep/wake cycles, neurobehavioral performance, and a wide range of diseases including metabolic, cardiovascular, and psychiatric disorders are impacted by these rhythms. Circadian clocks are present within individual cells and at tissue and organismal levels as emergent properties from the interaction of cellular oscillators. Mathematical models of circadian rhythms have been proposed to provide a better understanding of and to predict aspects of this complex physiological system. These models can be used to: (a) manipulate the system in silico with specificity that cannot be easily achieved using in vivo and in vitro experimental methods and at lower cost, (b) resolve apparently contradictory empirical results, (c) generate hypotheses, (d) design new experiments, and (e) to design interventions for altering circadian rhythms. Mathematical models differ in structure, the underlying assumptions, the number of parameters and variables, and constraints on variables. Models representing circadian rhythms at different physiologic scales and in different species are reviewed to promote understanding of these models and facilitate their use. This article is categorized under: Physiology > Mammalian Physiology in Health and Disease Models of Systems Properties and Processes > Organ, Tissue, and Physiological Models.


Spinal correction surgery improves asymmetrical trunk kinematics during gait in adolescent idiopathic scoliosis with thoracic major curve.
To clarify the effect of posterior correction and fusion surgery on the trunk-pelvic kinematics during gait in adolescent idiopathic scoliosis (AIS) patients with single thoracic major curve. Among preoperative AIS patients who planned correction surgery, 18 patients with Lenke type 1A or B were selected for this study. All patients were female. The patients' trunk and pelvic kinematics during gait were measured three-dimensionally and dynamically using reflective markers, optoelectronic motion capture system. The gait analysis was performed before and 1-2 years after surgery. The trunk and pelvic symmetry during gait was evaluated at coronal, sagittal, and transverse planes between concave and convex sides. The trunk and pelvic angles in sagittal and coronal planes were equivalent between concave and convex sides before and after surgery. Preoperatively, transverse trunk rotation angles were significantly deviated toward the concave (left) side during both static standing (4.3 ± 2.0°) and gait (8.8 ± 0.6°, p < 0.01). Preoperative transverse pelvic rotation angles were significantly deviated toward the convex side during static standing (4.0 ± 3.8°). However, pelvis displayed with symmetric rotational kinematics during gait. Postoperatively, the deviated transverse trunk rotation angle significantly decreased (1.6 ± 0.3°), and the transverse rotational kinematics of both trunk and pelvis improved to symmetric. Posterior correction and fusion surgery have improved preoperative asymmetric global rotational kinematics of trunk and pelvis in transverse plane to symmetric postoperatively in AIS patients with thoracic single major curve. These slides can be retrieved under Electronic Supplementary Material.


Genotyping and Quantifying Lyme Pathogen Strains by Deep Sequencing of the Outer Surface Protein C (ospC) Locus.
A mixed infection of a single tick or host by Lyme disease spirochetes is common and a unique challenge for the diagnosis, treatment, and surveillance of Lyme disease. Here, we describe a novel protocol for differentiating Lyme strains on the basis of deep sequencing of the hypervariable outer surface protein C locus (ospC). Improving upon the traditional DNA-DNA hybridization method, the next-generation sequencing-based protocol is high throughput, quantitative, and able to detect new pathogen strains. We applied the method to more than one hundred infected Ixodes scapularis ticks collected from New York State, USA, in 2015 and 2016. An analysis of strain distributions within individual ticks suggests an overabundance of multiple infections by five or more strains, inhibitory interactions among coinfecting strains, and the presence of a new strain closely related to Borreliella bissettiae A supporting bioinformatics pipeline has been developed. The newly designed pair of universal ospC primers target intergenic sequences conserved among all known Lyme pathogens. The protocol could be used for culture-free identification and quantification of Lyme pathogens in wildlife and potentially in clinical specimens.


Utilization of Lignin in Biopolymeric Packaging Films.
Lignin is a byproduct of agricultural industries and only has limited applications. In this study, lignin was investigated for use in sustainable biopolymeric packaging film. Alkali lignin (AL) and lignosulfonate (LSS) were added to enzymatically modified soy protein isolate (SPI) biopolymeric film with different concentrations with the goal of improvement of film physical and functional properties. A radical scavenging activity test revealed that films containing LSS had values 28 and 6% higher than control and AL-based films, respectively; AL itself (not in films) had significantly higher radical scavenging activity than LSS. This indicates the activity of lignin is affected by interaction with SPI. The higher compatibility between LSS and enzymatically modified SPI resulted in a positive effect on surface smoothness, water absorption, and mechanical properties of LSS-based films. Films containing AL showed a high light absorption range in the UV region, and this UV-blocking ability increased with increasing level of lignin. Deconvoluted Fourier transform infrared spectra confirmed that the addition of lignin resulted in some changes in the secondary structure of the protein matrix, which were aligned with X-ray diffraction results. The addition of lignin improved tensile strength (TS) and thermal stability of films compared to the lignin-free control. This improvement in TS and thermal stability was probably a result of new intermolecular interactions between lignin and SPI. Water vapor permeability of the films containing lignin decreased to 50% of the control because lignin played a role as a filler in the matrix. On the basis of our observations, the incorporation of lignin into biopolymeric film is capable of providing additional benefits and solutions to various industries, such as food, packaging, agriculture, and pharmaceuticals.


Water-in-Water Droplets by Passive Microfluidic Flow Focusing.
We present a simple microfluidic system that generates water-in-water, aqueous two phase system (ATPS) droplets, by passive flow focusing. ATPS droplet formation is achieved by applying weak hydrostatic pressures, with liquid-filled pipette tips as fluid columns at the inlets, to introduce low speed flows to the flow focusing junction. To control the size of the droplets, we systematically vary the interfacial tension and viscosity of the ATPS fluids and adjust the fluid column height at the fluid inlets. The size of the droplets scales with a power law of the ratio of viscous stresses in the two ATPS phases. Overall, we find a drop size coefficient of variation (CV; i.e., polydispersity) of about 10%. We also find that when drops form very close to the flow focusing junction, the drops have a CV of less than 1%. Our droplet generation method is easily scalable: we demonstrate a parallel system that generates droplets simultaneously and improves the droplet production rate by up to one order of magnitude. Finally, we show the potential application of our system for encapsulating cells in water-in-water emulsions by encapsulating microparticles and cells. To the best of our knowledge, our microfluidic technique is the first that forms low interfacial tension ATPS droplets without applying external perturbations. We anticipate that this simple approach will find utility in drug and cell delivery applications because of the all-biocompatible nature of the water-in-water ATPS environment.


CD81 is dispensable for hepatitis C virus cell-to-cell transmission in hepatoma cells.
Hepatitis C virus (HCV) infects cells by the direct uptake of cell-free virus following virus engagement with specific cell receptors such as CD81. Recent data have shown that HCV is also capable of direct cell-to-cell transmission, although the role of CD81 in this process is disputed. Here, we generated cell culture infectious strain JFH1 HCV (HCVcc) genomes carrying an alanine substitution of E2 residues W529 or D535 that are critical for binding to CD81 and infectivity. Co-cultivation of these cells with naïve cells expressing enhanced green fluorescent protein (EGFP) resulted in a small number of cells co-expressing both EGFP and HCV NS5A, showing that the HCVcc mutants are capable of cell-to-cell spread. In contrast, no cell-to-cell transmission from JFH1(DeltaE1E2)-transfected cells occurred, indicating that the HCV glycoproteins are essential for this process. The frequency of cell-to-cell transmission of JFH1(W529A) was unaffected by the presence of neutralizing antibodies that inhibit E2-CD81 interactions. By using cell lines that expressed little or no CD81 and that were refractive to infection with cell-free virus, we showed that the occurrence of viral cell-to-cell transmission is not influenced by the levels of CD81 on either donor or recipient cells. Thus, our results show that CD81 plays no role in the cell-to-cell spread of HCVcc and that this mode of transmission is shielded from neutralizing antibodies. These data suggest that therapeutic interventions targeting the entry of cell-free HCV may not be sufficient in controlling an ongoing chronic infection, but need to be complemented by additional strategies aimed at disrupting direct cell-to-cell viral transmission.


How do childhood intelligence and early psychosocial adversity influence income attainment among adult extremely low birth weight survivors? A test of the cognitive reserve hypothesis.
Perinatal and later postnatal adversities have been shown to adversely affect socioeconomic trajectories, while enhanced early cognitive abilities improve them. However, little is known about the combined influence of these exposures on social mobility. In this study, we examined if childhood IQ moderated the association between four different types of postnatal adversity (childhood socioeconomic disadvantage, childhood sexual abuse, lifetime psychiatric disorder, and trait neuroticism) and annual earnings at 30-35 years of age in a sample of 88 extremely low birth weight survivors. Our results suggested that higher childhood IQ was associated with greater personal income at age 30-35. Extremely low birth weight survivors who did not face psychological adversities and who had higher childhood IQ reported higher income in adulthood. However, those who faced psychological adversity and had higher childhood IQ generally reported lower income in adulthood. Our findings suggest that cognitive reserve may not protect preterm survivors against the complex web of risk factors affecting their later socioeconomic attainment.


Blockade of transgenic gamma delta T cell development in beta 2-microglobulin deficient mice.
The gamma delta T cell receptor (TCR) of the hybridoma KN6 recognizes the self molecule encoded by a class I gene which maps within the TL region of the major histocompatibility complex (MHC) of H-2b mice. Mice transgenic (Tg) for this TCR were crossed with mice genetically deficient in beta 2-microglobulin (beta 2m). No mature Tg gamma delta T cells were detected in the thymus or the spleen of the beta 2m- gamma delta Tg mice. We conclude that interaction between the Tg gamma delta TCR and a beta 2m-associated molecule (probably an MHC class I molecule) is required for the generation of mature Tg gamma delta T cells.


Zig-zag networks of self-excited periodic oscillations in a tunnel diode and a fiber-ring laser.
We report numerical evidence showing that periodic oscillations can produce unexpected and wide-ranging zig-zag parameter networks embedded in chaos in the control space of nonlinear systems. Such networks interconnect shrimplike windows of stable oscillations and are illustrated here for a tunnel diode, for an erbium-doped fiber-ring laser, and for the Hénon map, a proxy of certain CO(2) lasers. Networks in maps can be studied without the need for solving differential equations. Tuning parameters along zig-zag networks allows one to continuously modify wave patterns without changing their chaotic or periodic nature. In addition, we report convenient parameter ranges where such networks can be detected experimentally.


Bis(phenothiazine)arene diradicaloids: isolation, characterization and crystal structures.
A class of bis(phenothiazine)arene dications were isolated and structurally characterized by using weakly coordinating anions. Their electronic structures were investigated by various experiments in tandem with DFT calculations. They all feature singlet ground states with a considerable diradical character and may be regarded as nearly pure diradicals. Our work demonstrates the important role of orthogonal alignment of spin-carrying π-systems in tuning the electronic structures of bis(triarylamine) dications.


Coffee consumption and coronary heart disease risk using the Framingham risk score.
Although concerns regarding the influence of coffee consumption on human health have accompanied the massive increase in coffee consumption, the effects of coffee intake on the risk for coronary heart disease (CHD) remain controversial. Therefore, we evaluated the association between coffee consumption and CHD risk as estimated using the Framingham risk model in Korean adults. This cross-sectional study involved 3,987 participants aged 30-74 years who participated in the fifth Korea National Health and Nutrition Examination Survey conducted in 2010. The frequency of coffee consumption was self-reported and classified into 4 categories (non-drinker, 1, 2, and >=3 cups/day). The 10-year risk for CHD was determined from the Framingham risk score. Across the levels of coffee consumption, there were significant differences in the frequency of smoking among men and advanced age, low high-density lipoprotein cholesterol level, diabetes, and smoking among women. In the multiple logistic regression analyses, the adjusted odds ratios (95% CI) for >=20% 10-year CHD risk was 0.211 (0.060-0.745) for women who consumed >=3 cups of coffee per day compared with women who consumed <1 cup per day. For women, a significant dose-response inverse association between the level of coffee consumption and 10-year CHD risk persisted even after adjusting for multiple confounding factors. For the men, however, there was no significant association between coffee consumption and 10-year CHD risk. Coffee consumption is associated with a lower risk for CHD in Korean women.


Survey of the implementation of the recommendations in the Health Services Circular 1998/224 'Better Blood Transfusion'.
This report describes a questionnaire survey on the implementation of the recommendations of the Health Services Circular (HSC) 'Better Blood Transfusion' 1998/224 for improving transfusion practice. The survey was carried out to inform a second UK Chief Medical Officers' symposium on 'Better Blood Transfusion' in October 2001. Sixty-nine percent of hospitals where blood is transfused in England participated. The results show that, by 2001, most hospitals had established Hospital Transfusion Committees (HTCs), developed protocols for the process of transfusion and were participating in the Serious Hazards of Transfusion (SHOT) scheme. However, there was limited compliance with other recommendations, including the provision of training for staff involved in transfusion and information to patients, the development of protocols for the appropriate use of blood, the performance of audits of transfusion practice and the introduction of peri-operative cell salvage. The survey did not determine the reasons for this limited compliance. New initiatives including the issue of a further HSC on 'Better Blood Transfusion' are aimed at enabling hospitals to improve their transfusion practice in a more systematic way than that was found in the results of this survey.


Clinical development and regulatory points for consideration for second-generation live attenuated dengue vaccines.
Licensing and decisions on public health use of a vaccine rely on a robust clinical development program that permits a risk-benefit assessment of the product in the target population. Studies undertaken early in clinical development, as well as well-designed pivotal trials, allow for this robust characterization. In 2012, WHO published guidelines on the quality, safety and efficacy of live attenuated dengue tetravalent vaccines. Subsequently, efficacy and longer-term follow-up data have become available from two Phase 3 trials of a dengue vaccine, conducted in parallel, and the vaccine was licensed in December 2015. The findings and interpretation of the results from these trials released both before and after licensure have highlighted key complexities for tetravalent dengue vaccines, including concerns vaccination could increase the incidence of dengue disease in certain subpopulations. This report summarizes clinical and regulatory points for consideration that may guide vaccine developers on some aspects of trial design and facilitate regulatory review to enable broader public health recommendations for second-generation dengue vaccines.


Co-existence of scrapie prion protein types 1 and 2 in sporadic Creutzfeldt-Jakob disease: its effect on the phenotype and prion-type characteristics.
Five phenotypically distinct subtypes have been identified in sporadic Creutzfeldt-Jakob disease (sCJD), based on the methionine/valine polymorphic genotype of codon 129 of the prion protein (PrP) gene and the presence of either one of the two protease K-resistant scrapie prion protein (PrP(Sc)) types identified as 1 and 2. The infrequent co-existence of both PrP(Sc) types in the same case has been known for a long time. Recently, it has been reported, using type-specific antibodies, that the PrP(Sc) type 1 is present in all cases of sCJD carrying PrP(Sc) type 2. The consistent co-occurrence of both PrP(Sc) types complicates the diagnosis and the current classification of sCJD, and has implications for the pathogenesis of naturally occurring prion diseases. In the present study, we investigated the prevalence of PrP(Sc) types 1 and 2 co-occurrence, along with its effects on the disease phenotype and PrP(Sc) strain characteristics, comparatively analysing 34 cases of sCJD, all methionine homozygous at codon 129 of the PrP gene (sCJDMM). To minimize overestimating the prevalence of the sCJDMM cases carrying PrP(Sc) types 1 and 2 (sCJDMM1-2), we used proteinase K concentrations designed to hydrolyse all fragments resulting from an incomplete digestion, while preserving the protease-resistant PrP(Sc) core. Furthermore, we used several antibodies to maximize the detection of both PrP(Sc) types. Our data show that sCJDMM cases associated exclusively with either PrP(Sc) type 1 (sCJDMM1) or PrP(Sc) type 2 (sCJDMM2) do exist; we estimate that they account for approximately 56% and 5% of all the sCJDMM cases, respectively; while in 39% of the cases, both PrP(Sc) types 1 and 2 are present together (sCJDMM1-2) either mixed in the same anatomical region or separate in different regions. Clinically, sCJDMM1-2 had an average disease duration intermediate between the other two sCJDMM subtypes. The histopathology was also intermediate, except for the cerebellum where it resembled that of sCJDMM1. These features, along with the PrP immunostaining pattern, offer a diagnostic clue. We also observed a correlation between the disease duration and the prevalence of PrP(Sc) type 2 and sCJDMM2 phenotypes. The use of different antibodies and of the conformational stability immunoassay indicated that the co-existence of types 1 and 2 in the same anatomical region may confer special conformational characteristics to PrP(Sc) types 1 and 2. All of these findings indicate that sCJDMM1-2 should be considered as a separate entity at this time.


Depleting tumor-specific Tregs at a single site eradicates disseminated tumors.
Activation of TLR9 by direct injection of unmethylated CpG nucleotides into a tumor can induce a therapeutic immune response; however, Tregs eventually inhibit the antitumor immune response and thereby limit the power of cancer immunotherapies. In tumor-bearing mice, we found that Tregs within the tumor preferentially express the cell surface markers CTLA-4 and OX40. We show that intratumoral coinjection of anti-CTLA-4 and anti-OX40 together with CpG depleted tumor-infiltrating Tregs. This in situ immunomodulation, which was performed with low doses of antibodies in a single tumor, generated a systemic antitumor immune response that eradicated disseminated disease in mice. Further, this treatment modality was effective against established CNS lymphoma with leptomeningeal metastases, sites that are usually considered to be tumor cell sanctuaries in the context of conventional systemic therapy. These results demonstrate that antitumor immune effectors elicited by local immunomodulation can eradicate tumor cells at distant sites. We propose that, rather than using mAbs to target cancer cells systemically, mAbs could be used to target the tumor infiltrative immune cells locally, thereby eliciting a systemic immune response.


Genetic Susceptibility for Childhood BMI has no Impact on Weight Loss Following Lifestyle Intervention in Danish Children.
This study aimed to investigate the effect of a genetic risk score (GRS) comprising 15 single-nucleotide polymorphisms, previously shown to associate with childhood BMI, on the baseline cardiometabolic traits and the response to a lifestyle intervention in Danish children and adolescents. Children and adolescents with overweight or obesity (n = 920) and a population-based control sample (n = 698) were recruited. Anthropometric and biochemical measures were obtained at baseline and in a subgroup of children and adolescents with overweight or obesity again after 6 to 24 months of lifestyle intervention (n = 754). The effects of the GRS were examined by multiple linear regressions using additive genetic models. At baseline, the GRS associated with BMI standard deviation score (SDS) both in children and adolescents with overweight or obesity (β = 0.033 [SE = 0.01]; P = 0.001) and in the population-based sample (β = 0.065 [SE = 0.02]; P = 0.001). No associations were observed for cardiometabolic traits. The GRS did not influence changes in BMI SDS or cardiometabolic traits following lifestyle intervention. A GRS for childhood BMI was associated with BMI SDS but not with other cardiometabolic traits in Danish children and adolescents. The GRS did not influence treatment response following lifestyle intervention.


Effect of opioid receptor blockade on acetaldehyde self-administration and ERK phosphorylation in the rat nucleus accumbens.
We have previously shown that acetaldehyde (ACD), the first metabolite of ethanol, regulates its motivational properties and possesses reinforcing effects by itself. A large and still growing body of evidence indicates that the endogenous opioidergic system plays a critical role in the motivational effects of ethanol and suggests a role for extracellular signal-regulated kinase (ERK) in these effects of both ethanol and ACD. The present study was undertaken to examine if opioid-mediated mechanisms are involved in the reinforcing properties of ACD and in ACD-elicited ERK activation. To this end, Wistar rats were trained to orally self-administer ACD (0.2%) by nose poking. Responses on active nose poke caused delivery of ACD solution, whereas responses on inactive nose poke had no consequences. The effect of pretreatment with a nonselective opioid receptor antagonist, naltrexone (NTX), was evaluated during (1) maintenance of ACD self-administration, (2) deprivation effect after ACD extinction, and (3) ACD self-administration under a progressive-ratio schedule of reinforcement. Additionally, we tested the effect of NTX on saccharin (0.05%) reinforcement, as assessed by oral self-administration, and on ACD-elicited ERK phosphorylation in the nucleus accumbens (Acb), as assessed by immunohistochemistry. Finally, we examined the effect of a μ(1)-selective opioid receptor antagonist, naloxonazine (NLZ), on the maintenance phase of ACD and saccharin self-administration. The results indicate that NTX (0.4-0.8mg/kg) reduced the maintenance, the deprivation effect, and the break points of ACD self-administration without suppressing saccharin self-administration. Moreover, NTX decreased ACD-elicited ERK activation in the Acb shell and core. NLZ (10-15mg/kg) reduced the maintenance phase of ACD self-administration without interfering with saccharin self-administration, whereas both NTX and NLZ failed to modify responses on inactive nose poke indicating the lack of a nonspecific behavioral activation. Overall, these results indicate that the opioid system is implicated in the reinforcing properties of ACD and suggest an involvement of ERK. The finding that NTX and NLZ reduce ACD but not saccharin self-administration indicates that these effects are specific to ACD.


Xylo-oligosaccharides and virginiamycin differentially modulate gut microbial composition in chickens.
The emergence and spread of antibiotic resistance in pathogens have led to a restriction on the use of antibiotic growth promoters (AGPs) in animal feed in some countries. The potential negative after-effects of a ban on AGPs could be mitigated by improving animal intestinal health with prebiotic dietary fibers such as xylo-oligosaccharides (XOS). However, the mechanism(s) by which an antibiotic or prebiotic contributes to the health and growth of animals are not well understood. Here, we evaluated XOS and virginiamycin (VIRG)-mediated changes in gut microbiota of broiler chickens using pyrosequencing of the 16S rRNA gene. There was a significant change in the relative abundance of certain bacteria, but the overall microbial diversity was not affected by treatment with either XOS or VIRG. Supplementation of HXOS (2 g XOS/kg diet) increased the proportion of Lactobacillus genus in the cecum, whereas Propionibacterium and Corynebacterium genera were enriched in the ileum of VIRG (16 mg/kg) treated birds. Furthermore, an increase in the cecal concentrations of acetate and propionate was observed in HXOS- and VIRG-fed chickens, respectively. These two groups of birds had better feed conversion efficiencies in comparison with the control group from day 7 to 21. In addition, temporal variations in the gut microbiota were evident in the chickens of different ages. Treatments with XOS or VIRG modified the relative abundance but not the presence or absence of specific microbial genus. The increase in both Lactobacillus spp. and acetate production in the cecum of HXOS-treated chickens may promote intestinal health.


Multidisciplinary team working: collaboration and conflict.
The aim of this paper is to report on findings identifying some of the difficulties encountered by the multidisciplinary team in the development and implementation of a care pathway for patients diagnosed with schizophrenia. Policy direction has shifted towards greater team working and blurring of professional boundaries. Moreover, there is greater need to deliver care according to set standards and for patients to reach particular outcomes of care. Care pathways are relatively new for psychiatry and will potentially uncover tensions within the team. Data were collected by participant observation and semistructured interviews over a period of 12 months on an acute psychiatric unit. The care team developed the care pathway and the process of development and implementation was observed through action research. Cross-sectional indexing was used to analyse the data, and themes were developed using interview and observational data collection methods. Clinicians argued strongly for clear role boundaries but also defended their perceived control over health care from other professions. The findings indicate that designing a care pathway for people with schizophrenia may produce conflicting perceptions from the team. Conflict may arise through professions being unwilling to accept plurality over roles, which may hinder progress in meeting the needs of patients. The findings also counter the impression that care pathways can be implemented with little impact on the team.


Benefits of cardiac coherence combined with medical hypnosis on preoperative anxiety before cancer surgery: the COHEC II study trial protocol.
Preoperative anxiety is a frequent problem that can lead to complications both during anaesthesia and in the postoperative period, especially in oncology. Studies have shown that it can be managed using non-pharmacological approaches, but few works have evaluated psychoeducational programmes. The aim of the COHErence Cardiaque (COHEC) II Study is to evaluate the combination of medical hypnosis (MH) and cardiac coherence (CC) training to manage preoperative anxiety in patients with cancer. COHEC II is an ongoing multicentre randomised clinical trial carried out in three French comprehensive cancer centres. In total, 296 patients who will undergo surgery for cancer will be recruited during 18 months and will be randomised in the control arm or the intervention arm. Patients in the intervention arm will follow a daily programme that combines MH and CC, starting 7 days before surgery. The control arm will receive the standard treatment to manage preoperative anxiety. The primary endpoint is the anxiety level on surgery day, measured using a Visual Analogue Scale. Secondary endpoints are patient adherence to the programme, satisfaction and postsurgery recovery quality. The study protocol was approved by the French Ethics Committee (Comité de Protection des Personnes EST-II) on 24 November 2021 and will be carried out following the good practice guidelines and the Declaration of Helsinki. Results will be published in peer-reviewed journals and presented at conferences. NCT05197972.


Health-care professionals' perspective on discussing sexual issues in adult patients after haematopoietic cell transplantation.
The majority of adult patients have sexual concerns after post-haematopoietic cell transplantation. Even so, health-care professionals (HCP) do not routinely discuss these problems. We, therefore, surveyed all the members of the European Society for Blood and Marrow Transplantation to evaluate the barriers and facilitators to discussing sexual issues. The 73-item web-survey was completed by 166 registered nurses (RNs) and 126 medical doctors (MDs). Sixty-eight percent reported that they seldom discussed sexual issues. Younger MDs (p < 0.001) and those who work in non-western European countries (p = 0.003), RNs with probably less sexual education themselves (p = 0.002), MDs and RNs who have limited knowledge about sexual complications (p < 0.001) and MDs and RNs who feel uncomfortable discussing sexual issues (p < 0.001) are all less likely to discuss these matters. The major perceived barriers were that patients might be embarrassed if sexual issues were discussed in the presence of a relative (60% RNs, 67% MDs) and that professionals prefer patients to raise sexual issues themselves (54% RNs, 44% MDs). The most important perceived facilitator was for the patient to initiate discussion (≥ 90% for RNs and MDs). Overall, haematopoietic cell transplantation survivors may not be receiving the support on sexual issues they probably need.


Use of magnetic resonance imaging to detect occult spinal dysraphism in infants.
OBJECTIVE Cutaneous stigmata or congenital anomalies often prompt screening for occult spinal dysraphism (OSD) in asymptomatic infants. While a number of studies have examined the results of ultrasonography (US) screening, less is known about the findings when MRI is used as the primary imaging modality. The object of this study was to assess the results of MRI screening for OSD in infants. METHODS The authors undertook a retrospective review of all infants who had undergone MRI of the lumbar spine to screen for OSD over a 6-year period (September 2006-September 2012). All images had been obtained on modern MRI scanners using sequences optimized to detect OSD, which was defined as any fibrolipoma of the filum terminale (FFT), a conus medullaris ending at or below the L2-3 disc space, as well as more complex lesions such as lipomyelomeningocele (LMM). RESULTS Five hundred twenty-two patients with a mean age of 6.2 months at imaging were included in the study. Indications for imaging included isolated dimple in 235 patients (45%), asymmetrically deviated gluteal cleft in 43 (8%), symmetrically deviated (Y-shaped) gluteal cleft in 38 (7%), hemangioma in 28 (5%), other isolated cutaneous stigmata (subcutaneous lipoma, vestigial tail, hairy patch, and dysplastic skin) in 31 (6%), several of the above stigmata in 97 (18%), and congenital anomalies in 50 (10%). Twenty-three percent (122 patients) of the study population had OSD. Lesions in 19% of these 122 patients were complex OSD consisting of LMM, dermal sinus tract extending to the thecal sac, and lipomeningocele. The majority of OSD lesions (99 patients [81%]) were filar abnormalities, a group including FFT and low-lying conus. The rate of OSD ranged from 12% for patients with asymmetrically deviated gluteal crease to 55% for those with other isolated cutaneous stigmata. Isolated midline dimple was the most common indication for imaging. Among this group, 20% (46 of 235) had OSD. There was no difference in the rate of OSD based on dimple location. Those with OSD had a mean dimple position of 15 mm (SD 11.8) above the coccyx. Those without OSD had a mean dimple position of 12.2 mm (SD 19) above the coccyx (p = 0.25). CONCLUSIONS The prevalence of OSD identified with modern high-resolution MRI screening is significantly higher than that reported with US screening, particularly in patients with dimples. The majority of OSD lesions identified are FFT and low conus. The clinical significance of such lesions remains unclear.


Antihypertensive properties of a nitric oxide-releasing naproxen derivative in two-kidney, one-clip rats.
Nonsteroidal anti-inflammatory drugs have been reported to exacerbate hypertension. In this study, we tested the hypothesis that a nitric oxide-releasing derivative of naproxen would ameliorate hypertension in the rat. Hypertension was induced by partially occluding one renal artery (the "2K,1C" model), and 2 wk later the rats started receiving naproxen, the nitric oxide-releasing derivative HCT-3012, or vehicle each day for 2 wk. Naproxen significantly exacerbated the hypertension. HCT-3012 significantly reduced blood pressure relative to both the naproxen- and vehicle-treated groups. Both naproxen and HCT-3012 markedly suppressed whole blood thromboxane B(2) synthesis. In studies of anesthetized rats, naproxen significantly enhanced the late hypertensive response to endothelin-1 and significantly blunted the early hypotensive response. In contrast, HCT-3102 did not affect either response to endothelin-1. In vitro, HCT-3012 significantly reduced the responsiveness of aortic rings to the contractile effects of phenylephrine. These studies suggest that HCT-3012 reduces blood pressure in hypertensive rats, not simply through the vasodilatory actions of the nitric oxide it releases, but through alterations in the responsiveness of the vasculature to endogenous pressor agents.


Graphene-mediated microfluidic transport and nebulization via high frequency Rayleigh wave substrate excitation.
The deposition of a thin graphene film atop a chip scale piezoelectric substrate on which surface acoustic waves are excited is observed to enhance its performance for fluid transport and manipulation considerably, which can be exploited to achieve further efficiency gains in these devices. Such gains can then enable complete integration and miniaturization for true portability for a variety of microfluidic applications across drug delivery, biosensing and point-of-care diagnostics, among others, where field-use, point-of-collection or point-of-care functionality is desired. In addition to a first demonstration of vibration-induced molecular transport in graphene films, we show that the coupling of the surface acoustic wave gives rise to antisymmetric Lamb waves in the film which enhance molecular diffusion and hence the flow through the interstitial layers that make up the film. Above a critical input power, the strong substrate vibration displacement can also force the molecules out of the graphene film to form a thin fluid layer, which subsequently destabilizes and breaks up to form a mist of micron dimension aerosol droplets. We provide physical insight into this coupling through a simple numerical model, verified through experiments, and show several-fold improvement in the rate of fluid transport through the film, and up to 55% enhancement in the rate of fluid atomization from the film using this simple method.


Luminal influences on potassium secretion: chloride replacement with sulfate.
Electrolyte transport by the renal distal tubule of rats was studied by in vivo continuous microperfusion to determine whether replacing luminal chloride with sulfate would stimulate net potassium secretion. Results in a first series of experiments showed that replacing all Cl in the perfusion fluid with SO4 reduced net Na absorption, reversed the direction of net Cl transport from net absorption to net secretion, increased the lumen-negative transepithelial voltage, and increased net K secretion. In a second series of experiments adding NaCl to a perfusion fluid containing SO4 increased net Na absorption, reversed the direction of net Cl transport from net secretion to net absorption, decreased the transepithelial voltage, and decreased net K secretion. In a third series of experiments addition of 10(-6) M amiloride to a SO4-containing perfusion fluid reduced the transepithelial voltage but did not prevent the increase in K secretion, indicating that the changes in K secretion cannot be explained solely by changes in electrical driving forces. Since net fluid absorption was controlled, the changes in K secretion cannot be attributed to solvent drag effects. In all three series of experiments the rate of net K secretion was increased when luminal Cl concentration was reduced to less than 10 mM. The results suggest the existence of a cotransport system mediating K and Cl absorption by the distal tubule. Inhibition of K absorption by low lumen Cl may contribute to the increased net K secretion seen when nonchloride anions are present in distal fluid.


Circular proteins in plants: solution structure of a novel macrocyclic trypsin inhibitor from Momordica cochinchinensis.
Much interest has been generated by recent reports on the discovery of circular (i.e. head-to-tail cyclized) proteins in plants. Here we report the three-dimensional structure of one of the newest such circular proteins, MCoTI-II, a novel trypsin inhibitor from Momordica cochinchinensis, a member of the Cucurbitaceae plant family. The structure consists of a small beta-sheet, several turns, and a cystine knot arrangement of the three disulfide bonds. Interestingly, the molecular topology is similar to that of the plant cyclotides (Craik, D. J., Daly, N. L., Bond, T., and Waine, C. (1999) J. Mol. Biol. 294, 1327-1336), which derive from the Rubiaceae and Violaceae plant families, have antimicrobial activities, and exemplify the cyclic cystine knot structural motif as part of their circular backbone. The sequence, biological activity, and plant family of MCoTI-II are all different from known cyclotides. However, given the structural similarity, cyclic backbone, and plant origin of MCoTI-II, we propose that MCoTI-II can be classified as a new member of the cyclotide class of proteins. The expansion of the cyclotides to include trypsin inhibitory activity and a new plant family highlights the importance and functional variability of circular proteins and the fact that they are more common than has previously been believed. Insights into the possible roles of backbone cyclization have been gained by a comparison of the structure of MCoTI-II with the homologous acyclic trypsin inhibitors CMTI-I and EETI-II from the Cucurbitaceae plant family.


The Effect of Metoclopramide Addition to Lidocaine on Pain of Patients with Grades II and III Post-Episiotomy Repair.
Episiotomy is the most common procedure used for dilatation of the vaginal opening for giving birth. Although episiotomy is associated with benefits for the mother, it may lead to short term and long term disabilities, including postpartum perineal pain that is secondary to perineal tearing. The aim of this study was to investigate the effect of subcutaneous lidocaine compared with lidocaine and metoclopramide on pain after episiotomy. In this clinical trial study, the patients (83 for control group and 83 for case group) with episiotomy Grades II and III were enrolled and were asked to express their postpartum episiotomy pain as a number, from zero (no pain) to 10 (severest pain) Visual Analogue Scale (VAS score) at 0, 30 minute, 1 hour, 2 hour, 4 hour, 6 hour and 12 hour postpartum. Patients at the second stage of labour (crowning) were assigned to two groups: In the control group, 5 cc lidocaine 2% and in the case group, 5 cc lidocaine + 10 mg metoclopramide was injected for episiotomy. Then, the pain score was compared between the two groups. The data were analyzed by t-test and chi-square test with software SPSS version 20.0. Mean age of the women was 23.19±0.46 years in the control group and 23.96±0.58 years in the case group with no significant difference between the two groups (p<0.05). The mean pain score in the control group was 3.54±0.71 and in the case group 2.93±0.91 at 30 minutes after the injection, with a statistically significant difference. At other intervals, postnatal pain scores in the case group was lower than those of the control group (p<0.05). Injection of metoclopramide with lidocaine is more effective than lidocaine alone for relieving the pain after episiotomy.


The GARD platform for potency assessment of skin sensitizing chemicals.
Contact allergy induced by certain chemicals is a common health concern, and several alternative methods have been developed to fulfill the requirements of European legislation with regard to hazard assessment of potential skin sensitizers. However, validated methods, which provide information about the potency of skin sensitizers, are still lacking. The cell-based assay Genomic Allergen Rapid Detection (GARD), targeting key event 3, dendritic cell activation, of the skin sensitization AOP, uses gene expression profiling and a machine learning approach for the prediction of chemicals as sensitizers or non-sensitizers. Based on the GARD platform, we here expanded the assay to predict three sensitizer potency classes according to the European Classification, Labelling and Packaging (CLP) Regulation, targeting categories 1A (strong), 1B (weak) and no cat (non-sensitizer). Using a random forest approach and 70 training samples, a potential biomarker signature of 52 transcripts was identified. The resulting model could predict an independent test set consisting of 18 chemicals, six from each CLP category and all previously unseen to the model, with an overall accuracy of 78%. Importantly, the model was shown to be conservative and only underestimated the class label of one chemical. Furthermore, an association of defined chemical protein reactivity with distinct biological pathways illustrates that our transcriptional approach can reveal information contributing to the understanding of underlying mechanisms in sensitization.


Distinct mechanisms mediate speed-accuracy adjustments in cortico-subthalamic networks.
Optimal decision-making requires balancing fast but error-prone and more accurate but slower decisions through adjustments of decision thresholds. Here, we demonstrate two distinct correlates of such speed-accuracy adjustments by recording subthalamic nucleus (STN) activity and electroencephalography in 11 Parkinson's disease patients during a perceptual decision-making task; STN low-frequency oscillatory (LFO) activity (2-8 Hz), coupled to activity at prefrontal electrode Fz, and STN beta activity (13-30 Hz) coupled to electrodes C3/C4 close to motor cortex. These two correlates differed not only in their cortical topography and spectral characteristics but also in the relative timing of recruitment and in their precise relationship with decision thresholds. Increases of STN LFO power preceding the response predicted increased thresholds only after accuracy instructions, while cue-induced reductions of STN beta power decreased thresholds irrespective of instructions. These findings indicate that distinct neural mechanisms determine whether a decision will be made in haste or with caution.


Usual choline and betaine dietary intake and incident coronary heart disease: the Atherosclerosis Risk in Communities (ARIC) study.
Low dietary intake of the essential nutrient choline and its metabolite betaine may increase atherogenesis both through effects on homocysteine methylation pathways as well as through choline's antioxidants properties. Nutrient values for many common foods for choline and betaine have recently become available in the U.S. nutrient composition database. Our objective was to assess the association of dietary intake of choline and betaine with incident coronary heart disease (CHD), adjusting for dietary intake measurement error. We conducted a prospective investigation of the relation between usual intake of choline and betaine with the risk of CHD in 14,430 middle-aged men and women of the biethnic Atherosclerosis Risk in Communities study. A semi-quantitative food frequency questionnaire was used to assess nutrient intake. Proportional hazard regression models were used to calculate the risk of incident CHD. A regression calibration method was used to adjust for measurement error. During an average 14 years of follow-up (1987-2002), 1,072 incident CHD events were documented. Compared with the lowest quartile of intake, incident CHD risk was slightly and non-significantly higher in the highest quartile of choline and choline plus betaine, HR = 1.22 (0.91, 1.64) and HR = 1.14 (0.85, 1.53), controlling for age, sex, education, total energy intake, dietary intakes of folate, methionine and vitamin B6. No association was found between dietary choline intake and incident CHD when correcting for measurement error. Higher intakes of choline and betaine were not protective for incident CHD. Similar investigations in other populations are of interest.


Body mass index is associated with appropriateness of weight gain but not leisure-time physical activity during pregnancy.
Little is known about how leisure-time physical activity (LTPA) influences gestational weight gain (GWG) among body mass index (BMI) categories. The purpose of this study was to examine the relationship between pregnancy LTPA and the proportion of normal, overweight, and obese women who meet GWG recommendations. Participants included 449 subcohort women from the Pregnancy Outcomes and Community Health (POUCH) study. LTPA was collapsed into 3 categories [(None, < 7.5 kcal/kg/wk (low), ≥ 7.5 kcal/kg/wk (recommended)]. GWG was categorized according to IOM recommendations (low, recommended, or excess). Chi-square and logistic regression analyses were used to evaluate relationships among LTPA, BMI, and GWG. Overweight women were more likely to have high GWG vs. normal weight women (OR = 2.3, 95% CI 1.3-4.0). Obese women were more likely to experience low GWG (OR = 7.3, 95% CI 3.6-15.1; vs. normal and overweight women) or excess GWG (OR = 3.5, 95% CI 1.9-6.5; vs. normal weight women). LTPA did not vary by prepregnancy BMI category (P = .55) and was not related to GWG in any prepregnancy BMI category (P = .78). Regardless of prepregnancy BMI, LTPA did not affect a woman's GWG according to IOM recommendations. Results may be due to LTPA not differing among BMI categories.


Association of platelet-activating factor acetylhydrolase gene polymorphism with premature coronary artery disease in Turkish patients.
Platelet-activating factor (PAF) is a phospholipid with multiple actions that is involved in inflammatory diseases as well as in atherogenesis. It is inactivated by a plasma enzyme, PAF-acetylhydrolase (PAF-AH). Deficiency of this enzyme in plasma is caused by a missense mutation in the gene (G994T). The aim of this study was to investigate association of this mutation with premature coronary artery disease (CAD). One hundred and fifteen unrelated Turkish patients with a diagnosis of premature CAD and 128 unrelated healthy subjects were enrolled in this study. Genotyping was performed by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP). The prevalence of the G994T mutation in the patients was 2.60 % (heterozygote), and 0 % in the controls. There was no significant difference in allele frequency and genotype distribution among the study groups. The G9943T mutation in the plasma PAF acetylhydrolase gene is not associated with premature CAD in Turkish subjects.


PET imaging of epidermal growth factor receptor expression in tumours using 89Zr-labelled ZEGFR:2377 affibody molecules.
Epidermal growth factor receptor (EGFR) is a transmembrane tyrosine kinase receptor, which is overexpressed in many types of cancer. The use of EGFR-targeting monoclonal antibodies and tyrosine-kinase inhibitors improves significantly survival of patients with colorectal, non-small cell lung cancer and head and neck squamous cell carcinoma. Detection of EGFR overexpression provides important prognostic and predictive information influencing management of the patients. The use of radionuclide molecular imaging would enable non-invasive repeatable determination of EGFR expression in disseminated cancer. Moreover, positron emission tomography (PET) would provide superior sensitivity and quantitation accuracy in EGFR expression imaging. Affibody molecules are a new type of imaging probes, providing high contrast in molecular imaging. In the present study, an EGFR-binding affibody molecule (ZEGFR:2377) was site-specifically conjugated with a deferoxamine (DFO) chelator and labelled under mild conditions (room temperature and neutral pH) with a positron-emitting radionuclide (89)Zr. The (89)Zr-DFO-ZEGFR:2377 tracer demonstrated specific high affinity (160 ± 60 pM) binding to EGFR-expressing A431 epidermoid carcinoma cell line. In mice bearing A431 xenografts, (89)Zr-DFO-ZEGFR:2377 demonstrated specific uptake in tumours and EGFR-expressing tissues. The tracer provided tumour uptake of 2.6 ± 0.5% ID/g and tumour-to-blood ratio of 3.7 ± 0.6 at 24 h after injection. (89)Zr-DFO-ZEGFR:2377 provides higher tumour-to-organ ratios than anti-EGFR antibody (89)Zr-DFO-cetuximab at 48 h after injection. EGFR‑expressing tumours were clearly visualized by microPET using (89)Zr-DFO-ZEGFR:2377 at both 3 and 24 h after injection. In conclusion, 8(9)Zr-DFO-ZEGFR:2377 is a potential probe for PET imaging of EGFR-expression in vivo.


Cloning of DNA fragments from the left end of the adenovirus type 12 genome: transformation by cloned early region 1.
The human adenovirus serotype 12 (Ad-12) EcoRI-C DNA fragment (0 to 16.5 map units) was cloned in the plasmid vector pAT153. This cloned Ad-12 EcoRI-C DNA fragment was subcloned generating recombinant plasmids which contained the Ad-12 SalI-C fragment (0 to 10.3 map units), the Ad-12 HindIII-G fragment (0 to 6 X 8 map units) and the Ad-12 AccI-H fragment (0 to 4 X 7 map units). Thus, we constructed recombinant plasmids which contain Ad-12 DNA sequences which represent all or part of the virus transforming gene region. The capacity of the cloned Ad-12 EcoRI-C DNA fragment to transform rat cells in vitro was assessed using the focus assay on primary cultures of rat cells. The specific transforming activity of this recombinant plasmid was in the same range as that found for intact Ad-12 DNA. Transformed foci which were induced by the cloned Ad-12 EcoRI-C DNA fragment were established as cell lines and the presence of Ad-12 DNA in these lines was demonstrated using the Southern blotting technique.


Patients' experiences of chronic non-malignant musculoskeletal pain: a qualitative systematic review.
Musculoskeletal (MSK) pain is one of the most predominant types of pain and accounts for a large portion of the primary care workload. To systematically review and integrate the findings of qualitative research to increase understanding of patients' experiences of chronic non-malignant MSK pain. Synthesis of qualitative research using meta-ethnography using six electronic databases up until February 2012 (Medline, Embase, Cinahl, Psychinfo, Amed and HMIC). Databases were searched from their inception until February 2012, supplemented by hand-searching contents lists of specific journals for 2001-2011 and citation tracking. Full published reports of qualitative studies exploring adults' own experience of chronic non-malignant MSK pain were eligible for inclusion. Out of 24 992 titles, 676 abstracts, and 321 full texts were screened, 77 papers reporting 60 individual studies were included. A new concept of pain as an adversarial struggle emerged. This adversarial struggle was to: 1) affirm self; 2) reconstruct self in time; 3) construct an explanation for suffering; 4) negotiate the healthcare system; and 5) prove legitimacy. However, despite this struggle there is also a sense for some patients of 6) moving forward alongside pain. This review provides a theoretical underpinning for improving patient experience and facilitating a therapeutic collaborative partnership. A conceptual model is presented, which offers opportunities for improvement by involving patients, showing them their pain is understood, and forming the basis to help patients move forward alongside their pain.


Effects of heat stress and plane of nutrition on lactating Holstein cows: I. Production, metabolism, and aspects of circulating somatotropin.
Heat stress is detrimental to dairy production and affects numerous variables including feed intake and milk production. It is unclear, however, whether decreased milk yield is primarily due to the associated reduction in feed intake or the cumulative effects of heat stress on feed intake, metabolism, and physiology of dairy cattle. To distinguish between direct (not mediated by feed intake) and indirect (mediated by feed intake) effects of heat stress on physiological and metabolic indices, Holstein cows (n = 6) housed in thermal neutral conditions were pair-fed (PF) to match the nutrient intake of heat-stressed cows (HS; n = 6). All cows were subjected to 2 experimental periods: 1) thermal neutral and ad libitum intake for 9 d (P1) and 2) HS or PF for 9 d (P2). Heat-stress conditions were cyclical with daily temperatures ranging from 29.7 to 39.2 degrees C. During P1 and P2 all cows received i.v. challenges of epinephrine (d 6 of each period), and growth hormone releasing factor (GRF; d 7 of each period), and had circulating somatotropin (ST) profiles characterized (every 15 min for 6 h on d 8 of each period). During P2, HS cows were hyperthermic for the entire day and peak differences in rectal temperatures and respiration rates occurred in the afternoon (38.7 to 40.2 degrees C and 46 to 82 breaths/min, respectively). Heat stress decreased dry matter intake by greater than 35% and, by design, PF cows had similar reduced intakes. Heat stress and PF decreased milk yield, although the pattern and magnitude (40 and 21%, respectively) differed between treatments. The reduction in dry matter intake caused by HS accounted for only approximately 35% of the decrease in milk production. Both HS and PF cows entered into negative energy balance, but only PF cows had increased (approximately 120%) basal nonesterified fatty acid (NEFA) concentrations. Both PF and HS cows had decreased (7%) plasma glucose levels. The NEFA response to epinephrine did not differ between treatments but was increased (greater than 50%) in all cows during P2. During P2, HS (but not PF) cows had a modest reduction (16%) in plasma insulin-like growth factor-I. Neither treatment nor period had an effect on the ST response to GRF and there was little or no treatment effect on mean ST levels or pulsatility characteristics, but both HS and PF cows had reduced mean ST concentrations during P2. In summary, reduced nutrient intake accounted for just 35% of the HS-induced decrease in milk yield, and modest changes in the somatotropic axis may have contributed to a portion of the remainder. Differences in basal NEFA between PF and HS cows suggest a shift in postabsorptive metabolism and nutrient partitioning that may explain the additional reduction in milk yield in cows experiencing a thermal load.


Certified Reference Materials and their need for the diagnosis of autoimmune diseases.
Autoantibody measurement is the chosen tool, in addition to clinical observations, for the diagnosis of autoimmune diseases. Hence, it is essential for these measurements to be reliable and in the longer run to be standardised. Due to the intrinsic variability of analytes and reagents, and the heterogeneity of the available techniques, standardisation cannot be taken for granted, and results may vary between laboratories. As a consequence, diagnoses can be missed or wrong and unnecessary costs may burden individuals and healthcare systems. Standardisation of autoantibody testing is a demanding and multi-parameter task, but could be part of the solution. So as to achieve standardisation, the development and availability of suitable certified reference materials for calibration and/or quality control is crucial.


Alcohol brief intervention in community pharmacies: a feasibility study of outcomes and customer experiences.
Studies indicate that community pharmacy-based alcohol brief intervention (BI) is feasible. However, few studies report significant reductions in post-BI alcohol consumption and customer experience. Cost-effectiveness has not been previously examined. This 5 month study adopted a single group pre- and post-experimental design to: (1) assess uptake of the community pharmacy alcohol BI service; (2) establish post-BI changes in alcohol consumption for hazardous drinkers; (3) report the acceptability of the service to customers who received it; and (4) undertake a preliminary economic evaluation of the service through establishing whether pharmacy-based alcohol BI affected health and social care costs, including lost employment costs, and whether it was cost-effective. 26 community pharmacies in south London, UK. Trained pharmacists used the AUDIT-C and a retrospective 7-day Drinking Diary to identify risky drinkers and inform feedback and advice. Harmful drinkers were referred to their general practitioner and/or specialist alcohol services. A confidential service feedback questionnaire was completed by alcohol BI recipients. Baseline and 3-month follow-up telephone interviews were conducted with hazardous and low risk drinkers to assess post-BI alcohol use change and service cost-effectiveness. AUDIT-C, 7-day alcohol unit consumption, drinking days, cost utilisation data. Of the 663 eligible customers offered alcohol BI, 141 (21 %) took up the service. Three-quarters of customers were identified as risky drinkers. Follow-up interviews were conducted with 61 hazardous/low risk drinkers (response rate = 58 %). Hazardous drinkers were found to significantly reduce their 7-day alcohol unit consumption and drinking days, but not AUDIT-C scores. The majority of harmful drinkers (91 %, n = 10) who were contactable post-BI had accessed further alcohol related services. Customer feedback was generally positive. Over 75 % of customers would recommend the service to others. The cost of delivering the service was estimated to be £ 134. The difference in service costs pre-BI and post-BI was not statistically significant and remained non-significant when calculated on 500 customers receiving the intervention. Community pharmacy-based alcohol BI is a low cost service that may not have immediate beneficial impact on health and social service use, but can be effective in reducing drinking in hazardous drinkers.


Corticospinal activation of internal oblique muscles has a strong ipsilateral component and can be lateralised in man.
Trunk muscles receive corticospinal innervation ipsilaterally and contralaterally and here we investigate the degree of ipsilateral innervation and any cortical asymmetry in pairs of trunk muscles and proximal and distal limb muscles. Transcranial magnetic stimulation (TMS) was applied to left and right motor cortices in turn and bilateral electromyographic (EMG) recordings were made from internal oblique (IO; lower abdominal), deltoid (D; shoulder) and first dorsal interosseus (1DI; hand) muscles during voluntary contraction in ten healthy subjects. We used a 7-cm figure-of-eight stimulating coil located 2 cm lateral and 2 cm anterior to the vertex over either cortex. Incidence of ipsilateral motor evoked potentials (MEPs) was 85% in IO, 40% in D and 35% in 1DI. Mean (+/- S.E.M.) ipsilateral MEP latencies were longer ( P<0.05; paired t-test) than contralateral MEP latencies (contralateral vs. ipsilateral; IO: 16.1+/-0.4 ms vs. 19.0+/-0.5 ms; D: 9.7+/-0.3 ms vs. 15.1+/-1.9 ms; 1DI: 18.3+/-0.6 ms vs. 23.3+/-1.4 ms), suggesting that ipsilateral MEPs were not a result of interhemispheric current spread. Where data were available, we calculated a ratio (ipsilateral MEP areas/contralateral MEP areas) for a given muscle (IO: n=16; D: n=8; 1DI: n=7 ratios). Mean values for these ratios were 0.70+/-0.20 (IO), 0.14+/-0.05 (D) and 0.08+/-0.02 (1DI), revealing stronger ipsilateral drive to IO. Comparisons of the sizes of these ratios revealed a bias towards one cortex or the other (four subjects right; three subjects left). The predominant cortex showed a mean ratio of 1.21+/-0.38 compared with 0.26+/-0.06 in the other cortex ( P<0.05). It appears that the corticospinal control of IO has a strong ipsilateral component relative to the limb muscles and also shows hemispheric asymmetry.


Droplet evaporation on heated hydrophobic and superhydrophobic surfaces.
The evaporation characteristics of sessile water droplets on smooth hydrophobic and structured superhydrophobic heated surfaces are experimentally investigated. Droplets placed on the hierarchical superhydrophobic surface subtend a very high contact angle (∼160°) and demonstrate low roll-off angle (∼1°), while the hydrophobic substrate supports corresponding values of 120° and ∼10°. The substrates are heated to different constant temperatures in the range of 40-60 °C, which causes the droplet to evaporate much faster than in the case of natural evaporation without heating. The geometric parameters of the droplet, such as contact angle, contact radius, and volume evolution over time, are experimentally tracked. The droplets are observed to evaporate primarily in a constant-contact-angle mode where the contact line slides along the surface. The measurements are compared with predictions from a model based on diffusion of vapor into the ambient that assumes isothermal conditions. This vapor-diffusion-only model captures the qualitative evaporation characteristics on both test substrates, but reasonable quantitative agreement is achieved only for the hydrophobic surface. The superhydrophobic surface demonstrates significant deviation between the measured evaporation rate and that obtained using the vapor-diffusion-only model, with the difference being amplified as the substrate temperature is increased. A simple model considering thermal diffusion through the droplet is used to highlight the important role of evaporative cooling at the droplet interface in determining the droplet evaporation characteristics on superhydrophobic surfaces.


Contemporary Multi-Institutional Cohort of 550 Cases of Phyllodes Tumors (2007-2017) Demonstrates a Need for More Individualized Margin Guidelines.
Phyllodes tumors (PTs) are rare breast neoplasms, which have little granular data on margins. Current guidelines recommend ≥ 1 cm margins; however, recent data suggest narrower margins are sufficient, and for benign PT, a negative margin may not be necessary. We performed an 11-institution contemporary (2007-2017) review of PT practices. Demographics, surgical, and histopathologic data were captured. Logistic regression was used to estimate the association of select covariates with local recurrence (LR). Of 550 PT patients, the majority underwent excisional biopsy (55.3%, n = 302/546) or lumpectomy (wide excision) (38.5%, n = 210/546). Median tumor size was 30 mm, 68.9% (n = 379) were benign, 19.6% (n = 108) borderline, and 10.5% (n = 58) malignant. Surgical margins were positive in 42% (n = 231) and negative in 57.3% (n = 311). A second operation was performed in 38.0% (n = 209) of the total cohort, including 51 patients with an initial negative margin (82.4% with < 2 mm), and 157 with an initial positive margin, with residual disease only found in six (2.9%). Notably, 32.0% (n = 74) of those with an initial positive margin did not undergo a second operation, among whom only 2.7% (n = 2) recurred. Recurrence occurred in 3.3% (n = 18) of the total cohort (n = 15 LR, n = 3 distant), at median follow-up of 36.7 months. LR (all PT grades) was not reduced with wider negative margin width (≥ 2 mm v < 2 mm: odds ratio [OR] = 0.39; 95% CI, 0.07 to 2.10; P = .27) or final margin status (positive v negative: OR = 0.96; 95% CI, 0.26 to 3.52; P = .96). In current practice, many patients are managed outside of current guidelines. For the entire cohort, a wider margin width was not associated with a reduced risk of LR. We do not recommend re-excision of a negative margin for benign PT, regardless of margin width, as a progressively wider surgical margin is unlikely to reduce LR.


Pancreatic Cancer Metabolism: Breaking It Down to Build It Back Up.
How do cancer cells escape tightly controlled regulatory circuits that link their proliferation to extracellular nutrient cues? An emerging theme in cancer biology is the hijacking of normal stress response mechanisms to enable growth even when nutrients are limiting. Pancreatic ductal adenocarcinoma (PDA) is the quintessential aggressive malignancy that thrives in nutrient-poor, hypoxic environments. PDAs overcome these limitations through appropriation of unorthodox strategies for fuel source acquisition and utilization. In addition, the interplay between evolving PDA and whole-body metabolism contributes to disease pathogenesis. Deciphering how these pathways function and integrate with one another can reveal novel angles of therapeutic attack. Alterations in tumor cell and systemic metabolism are central to the biology of pancreatic cancer. Further investigation of these processes will provide important insights into how these tumors develop and grow, and suggest new approaches for its detection, prevention, and treatment.


Assessing interpersonal and communication skills in emergency medicine.
Interpersonal and communication skills (ICS) are a key component of several competency-based schemata and key competency in the set of six Accreditation Council for Graduate Medical Education (ACGME) core competencies. With the shift toward a competency-based educational framework, the importance of robust learner assessment becomes paramount. The journal Academic Emergency Medicine (AEM) hosted a consensus conference to discuss education research in emergency medicine (EM). This article summarizes the initial preparatory research that was conducted to brief consensus conference attendees and reports the results of the consensus conference breakout session as it pertains to ICS assessment of learners. The goals of this consensus conference session were to twofold: 1) to determine the state of assessment of observable learner performance and 2) to determine a research agenda within the ICS field for medical educators. The working group identified six key recommendations for medical educators and researchers.


Multiparticle biased diffusion-limited aggregation with surface diffusion: A comprehensive model of electrodeposition
We present a complete study of the multiparticle biased diffusion-limited aggregation (MBDLA) model supplemented with surface diffusion (SD), focusing on the relevance and effects of the latter transport mechanism. By comparing different algorithms, we show that MBDLA + SD is a very good qualitative model for electrodeposition in essentially the whole range of current intensities provided one introduces SD in the model in the proper fashion. We have found that the correct procedure involves simultaneous bulk diffusion and SD, introducing a time scale arising from the ratio of the rates of the two processes. We discuss in detail the different morphologies obtained and compare them to the available experimental data with very satisfactory results. We also characterize the aggregates thus obtained by means of the dynamic scaling exponents of the interface height, allowing us to distinguish several regimes in the mentioned interface growth. Our asymptotic scaling exponents are again in good agreement with recent experiments. We conclude by discussing a global picture of the influence and consequences of SD in electrodeposition.


Chronic and acute effects of walnuts on antioxidant capacity and nutritional status in humans: a randomized, cross-over pilot study.
Compared with other common plant foods, walnuts (Juglans regia) are consistently ranked among the highest in antioxidant capacity. In vitro, walnut polyphenols inhibit plasma and LDL oxidation, while in animal models they lower biomarkers of oxidative stress and raise antioxidant capacity. A limited number of human feeding trials indicate that walnuts improve some measures of antioxidant status, but not others. A 19 wk, randomized crossover trial was conducted in 21 generally healthy men and postmenopausal women > or = 50 y to study the dose-response effects of walnut intake on biomarkers of antioxidant activity, oxidative stress, and nutrient status. Subjects were randomized to receive either 21 or 42 g raw walnuts/d during each 6 wk intervention phase with a 6 wk washout between phases. Subjects were instructed to consume their usual diet, but refrain from eating any other tree nuts, seeds, peanuts, or ellagitannin-rich foods during the entire study, and other polyphenol-rich foods for 2 d prior to each study visit. Compared to baseline levels, red blood cell (RBC) linoleic acid and plasma pyridoxal phosphate (PLP) were significantly higher after 6 wk with 42 g/d walnuts (P < 0.05 for both). Overall, changes in plasma total thiols, and other antioxidant biomarkers, were not significant with either walnut dose. However, when compared to fasting levels, plasma total thiols were elevated within 1 h of walnut consumption with both doses during the baseline and end visits for each intervention phase (P < 0.05 for all). Despite the observed increase in RBC linoleic and linolenic acids associated with walnut consumption, this substrate for lipid peroxidation only minimally affected malondialdehyde (MDA) and antioxidant capacity. The proportional changes in MDA and Oxygen Radical Absorbance Capacity (ORAC) were consistent with a dose-response effect, although no significant within- or between-group differences were observed for these measures. Walnut consumption did not significantly change the plasma antioxidant capacity of healthy, well-nourished older adults in this pilot study. However, improvements in linoleic acid and pyridoxal phosphate were observed with chronic consumption, while total plasma thiols were enhanced acutely. Future studies investigating the antioxidant effects of walnuts in humans are warranted, but should include either a larger sample size or a controlled feeding intervention. ClinicalTrials.gov: NCT00626691.


A phase III, multicenter, randomized, placebo-controlled clinical trial of topical aminocaproic acid (Caprogel) in the management of traumatic hyphema.
To determine the safety and efficacy of topical aminocaproic acid (Caprogel) in the management of traumatic hyphema. Multicenter, randomized, double-masked, placebo-controlled clinical trial. A total of 51 patients participated in this trial (power = 36%, 2-tailed test). Patients presenting with traumatic hyphema were randomly assigned to 5-day treatment with topical aminocaproic acid or a placebo gel. Patients were monitored daily with ocular examination and vital sign testing for the 5 days of treatment and at 24 and 48 hours after treatment. General physical examination and laboratory testing were performed at baseline and day 5. The main efficacy variable was the rate of rebleeding. Secondary efficacy variables included time to hyphema clearance, intraocular pressure, time to secondary hemorrhage, and visual acuity. Safety variables included adverse events, vital signs, and laboratory measurements. Rebleeding occurred in 30% of the placebo group (8 of 27; 95% confidence interval [CI] = 14-50%), versus 8% of the treatment group (2 of 24; 95% CI = 1-27%), for an estimated continuity-corrected difference in percentage of patients with bleeding of 17% (95% CI = -3-38%). Secondary efficacy variables were similar in the groups, except that there was a trend towards more visual improvement in the topical aminocaproic acid group (54%) than in the placebo group (30%) at the last measurement (P = 0.08). Adverse events were similar. This study provides evidence that topical aminocaproic acid is safe and demonstrates trends towards reducing the rebleeding rate in the management of traumatic hyphema. However, because the study was terminated before complete enrollment, more definitive recommendations will require a larger trial.


Diabetes symptoms predictors of health-related quality of life in adolescents and young adults with type 1 or type 2 diabetes.
The objective was to investigate the patient-reported diabetes symptoms predictors of generic health-related quality of life (HRQOL) in adolescents and young adults (AYA) with type 1 or type 2 diabetes. The 15-item PedsQL™ 3.2 Diabetes Module Diabetes Symptoms Summary Score and PedsQL™ 4.0 Generic Core Scales were completed in a 10-site national field test study by 513 AYA ages 13-25 years with type 1 (n = 424) or type 2 (n = 89) diabetes. Diabetes symptoms were tested for bivariate and multivariate linear associations with generic HRQOL. Diabetes symptoms were associated with decreased HRQOL in bivariate analyses. In predictive analytics models utilizing hierarchical multiple regression analyses controlling for relevant demographic and clinical covariates, diabetes symptoms accounted for 38 and 39% of the variance in patient-reported generic HRQOL for type 1 and type 2 diabetes, respectively, reflecting large effect sizes. The diabetes symptoms facets hyperglycemia symptoms, hypoglycemia symptoms, and nonspecific diabetes symptoms individually accounted for a significant percentage of the variance in separate exploratory predictive analytics models after controlling for demographic and clinical covariates, with small-to-large effect sizes. Diabetes symptoms are potentially modifiable predictors of generic HRQOL in AYA with diabetes. Identifying specific diabetes symptoms or symptoms facets that are the most important predictors from the patient perspective facilitates a patient-centered approach in clinical research, clinical trials, and practice designed to enhance overall generic HRQOL in AYA with diabetes.


Naloxone alters the early response to an inspiratory flow-resistive load.
In a previous study in unanesthetized goats, we demonstrated that cerebrospinal fluid levels of beta-endorphin were significantly elevated after 2.5 h of inspiratory flow-resistive loading. Naloxone (NLX) (0.1 mg/kg) administration partially and transiently reversed the tidal volume depression seen during loading. In the current study, we tested the hypothesis that endogenous opioid elaboration results in depression of respiratory output to the diaphragm. In six studies of five unanesthetized goats, tidal volume (VT), transdiaphragmatic pressure (Pdi), diaphragmatic electromyogram (EMGdi), and arterial blood gases were monitored. A continuous NLX (0.1 mg/kg) or saline (SAL) infusion was begun 5 min before an inspiratory flow-resistive load of 120 cmH2O.l-1.s was imposed. Our data show that the depression of VT induced by the load was prevented by NLX as early as 15 min and persisted for 2 h. At 2 h, Pdi was still 294 +/- 45% of the base-line value compared with 217 +/- 35% during SAL. There was no difference in EMGdi between the groups at any time. However, the augmentation of Pdi was associated with a greater increase in end-expiratory gastric pressure in the NLX group. We conclude that the reduction in VT and Pdi associated with endogenous opioid elaboration is not mediated by a decrease in neural output to the diaphragm, but it appears to be the result of a decrease in respiratory output to the abdominal muscles.


Multi-stimuli-responsive supramolecular gel constructed by pillar[5]arene-based pseudorotaxanes for efficient detection and separation of multi-analytes in aqueous solution.
Here, a novel pseudorotaxanes-type crosslinker of a supramolecular polymer network (WP5-PN) has been constructed from a host water-soluble pillar[5]arene (WP5) and a guest naphthalene dimethylamine derivative (PN) via a stepwise process involving multiple non-covalent interactions. The obtained supramolecular polymers were able to transform into a supramolecular polymer gel (WP5-PN-G) and show AIE properties in DMSO-H2O binary solution. Interestingly, due to the dynamic and reversible nature of non-covalent interactions, the resultant supramolecular polymer gels exhibited external stimuli-responsiveness to different parameters, such as temperature, acid-base, competitive guest and mechanical stress. Moreover, WP5-PN-G showed fluorescent response for Fe3+ and Cu2+, while its xerogel showed excellent recyclable separation properties for these metal ions with adsorption rates up to 98.07% and 95.38%, respectively. Moreover, by rational introduction of these metal ions into the WP5-PN-G, corresponding metal ion coordinated metallogels, such as WP5-PN-FeG and WP5-PN-CuG were obtained. These metallogels could selectively and sensitively sense F- and CN-, respectively. The detection limits of these metallogels for F- and CN- were about 1 × 10-8 M. The WP5-PN-G has potential applications in multi-analytes detection and separation as well as fluorescent display materials.


Cingulo-insular structural alterations associated with psychogenic symptoms, childhood abuse and PTSD in functional neurological disorders.
Adverse early-life events are predisposing factors for functional neurological disorder (FND) and post-traumatic stress disorder (PTSD). Cingulo-insular regions are implicated in the biology of both conditions and are sites of stress-mediated neuroplasticity. We hypothesised that functional neurological symptoms and the magnitude of childhood abuse would be associated with overlapping anterior cingulate cortex (ACC) and insular volumetric reductions, and that FND and PTSD symptoms would map onto distinct cingulo-insular areas. This within-group voxel-based morphometry study probes volumetric associations with self-report measures of functional neurological symptoms, adverse life events and PTSD symptoms in 23 mixed-gender FND patients. Separate secondary analyses were also performed in the subset of 18 women with FND to account for gender-specific effects. Across the entire cohort, there were no statistically significant volumetric associations with self-report measures of functional neurological symptom severity or childhood abuse. In women with FND, however, parallel inverse associations were observed between left anterior insular volume and functional neurological symptoms as measured by the Patient Health Questionnaire-15 and the Screening for Somatoform Symptoms Conversion Disorder subscale. Similar inverse relationships were also appreciated between childhood abuse burden and left anterior insular volume. Across all subjects, PTSD symptom severity was inversely associated with dorsal ACC volume, and the magnitude of lifetime adverse events was inversely associated with left hippocampal volume. This study reveals distinct cingulo-insular alterations for FND and PTSD symptoms and may advance our understanding of FND. Potential biological convergence between stress-related neuroplasticity, functional neurological symptoms and reduced insular volume was identified.


Dielectric spectroscopy platform to measure MCF10A epithelial cell aggregation as a model for spheroidal cell cluster analysis.
3-Dimensional cell cultures are more representative of the native environment than traditional cell cultures on flat substrates. As a result, 3-dimensional cell cultures have emerged as a very valuable model environment to study tumorigenesis, organogenesis and tissue regeneration. Many of these models encompass the formation of cell aggregates, which mimic the architecture of tumor and organ tissue. Dielectric impedance spectroscopy is a non-invasive, label free and real time technique, overcoming the drawbacks of established techniques to monitor cell aggregates. Here we introduce a platform to monitor cell aggregation in a 3-dimensional extracellular matrix using dielectric spectroscopy. The MCF10A breast epithelial cell line serves as a model for cell aggregation. The platform maintains sterile conditions during the multi-day assay while allowing continuous dielectric spectroscopy measurements. The platform geometry optimizes dielectric measurements by concentrating cells within the electrode sensing region. The cells show a characteristic dielectric response to aggregation which corroborates with finite element analysis computer simulations. By fitting the experimental dielectric spectra to the Cole-Cole equation, we demonstrated that the dispersion intensity Δε and the characteristic frequency fc are related to cell aggregate growth. In addition, microscopy can be performed directly on the platform providing information about cell position, density and morphology. This platform could yield many applications for studying the electrophysiological activity of cell aggregates.


Neuroprotection by Estrogen and Progesterone in Traumatic Brain Injury and Spinal Cord Injury.
In recent years there has been a growing body of clinical and laboratory evidence demonstrating the neuroprotective effects of estrogen and progesterone after traumatic brain injury (TBI) and spinal cord injury (SCI). In humans, women have been shown to have a lower incidence of morbidity and mortality after TBI compared with age-matched men. Similarly, numerous laboratory studies have demonstrated that estrogen and progesterone administration is associated with a mortality reduction, improvement in neurological outcomes, and a reduction in neuronal apoptosis after TBI and SCI. Here, we review the evidence that supports hormone-related neuroprotection and discuss possible underlying mechanisms. Estrogen and progesterone-mediated neuroprotection are thought to be related to their effects on hormone receptors, signaling systems, direct antioxidant effects, effects on astrocytes and microglia, modulation of the inflammatory response, effects on cerebral blood flow and metabolism, and effects on mediating glutamate excitotoxicity. Future laboratory research is needed to better determine the mechanisms underlying the hormones' neuroprotective effects, which will allow for more clinical studies. Furthermore, large randomized clinical control trials are needed to better assess their role in human neurodegenerative conditions.


H1 linker histone promotes epigenetic silencing by regulating both DNA methylation and histone H3 methylation.
Epigenetic silencing in mammals involves DNA methylation and posttranslational modifications of core histones. Here we show that the H1 linker histone plays a key role in regulating both DNA methylation and histone H3 methylation at the H19 and Gtl2 loci in mouse ES cells. Some, but not all, murine H1 subtypes interact with DNA methyltransferases DNMT1 and DNMT3B. The interactions are direct and require a portion of the H1 C-terminal domain. Expression of an H1 subtype that interacts with DNMT1 and DNMT3B in ES cells leads to their recruitment and DNA methylation of the H19 and Gtl2 imprinting control regions. H1 also interferes with binding of the SET7/9 histone methyltransferase to the imprinting control regions, inhibiting production of an activating methylation mark on histone H3 lysine 4. H1-dependent recruitment of DNMT1 and DNMT3B and interference with the binding of SET7/9 also were observed with chromatin reconstituted in vitro. The data support a model in which H1 plays an active role in helping direct two processes that lead to the formation of epigenetic silencing marks. The data also provide evidence for functional differences among the H1 subtypes expressed in somatic mammalian cells.


Patient satisfaction compared with general health and disease-specific questionnaires in knee arthroplasty patients.
When assessing the health status of patients after orthopaedic surgery, such as knee arthroplasty, general health and disease-specific questionnaires are gaining in popularity because of their precision in detecting subtle differences. Self-administered postal surveys using extensive questionnaires have associated patient burden, however, which may have an impact on response rate and completeness. When a high response rate is important or when the use of comprehensive questionnaires is not practical, it may be possible to gain useful outcome data after a surgical procedure by simpler means. Two postal surveys to knee arthroplasty patients were performed. In the first survey, we posed a simple question regarding patient satisfaction to 27,114 patients. A second survey was sent 9 months later to 3,600 of the same patients; the same simple satisfaction question was posed along with several previously validated general health (NHP, SF36, SF12) and disease/site-specific (Oxford-12, WOMAC) outcome questionnaires. We found that patient satisfaction correlates significantly with general health and disease-specific outcome measures, with the highest correlation to the domains that relate to pain and function. When sent a simple satisfaction questionnaire, 95% of the patients answered, whereas the usable return rate of the more comprehensive questionnaires was 18% to 45% lower. Patients not responding to the comprehensive questionnaires were more often unsatisfied with their operated knee than patients responding.


Prospective evaluation of glutamine and phospholipids levels in first degree relatives of patients with Type 1 Diabetes from a multiethnic population.
A dysregulation in the metabolism of lipids may be an early marker of autoimmunity in Type 1 Diabetes (T1D). It would be of general importance to identify metabolic patterns that would predict the risk for T1D later in life. The aim of this study was to perform a prospective evaluation of glutamine and phospholipids levels in Brazilian first degree relatives (FDR) of patients with T1D in a mean interval of 5 years. Brazilian FDR (n = 30) of patients with T1D were evaluated and blood was sampled to measure the levels of glutamine and phospholipids in the fasting serum by quantitative colorimetric method. The tests were repeated after a mean interval of 5 years and compared to a control group (n = 20). The FDR presented lower levels of phospholipids than controls (p = 0.028), but not of glutamine (p = 0.075). Phospholipids levels decreased over time (p = 0.028) in FDR and were associated with Glutamic acid decarboxylase autoantibody (GADA) titers (p = 0.045), autoantibody positivity (p = 0.008) and PTPN22 polymorphisms (p = 0.014). In this Brazilian multiethnic population, there was a significant decrease in phospholipids levels in FDR in patients with T1D during a 5-year prospective follow-up, as well as a significant association between these metabolite, GADA and PTPN22 polymorphisms. For Glutamine no difference was found. These findings suggest that a dysregulation in the metabolism of lipids may precede the onset of the autoimmunity in T1D.


Novel phenotypes of Escherichia coli tat mutants revealed by global gene expression and phenotypic analysis.
The Tat protein export system serves to export folded proteins harboring an N-terminal twin arginine signal peptide across the cytoplasmic membrane. In this study, we have used gene expression profiling of Escherichia coli supported by phenotypic analysis to investigate how cells respond to a defect in the Tat pathway. Previous work has demonstrated that strains mutated in genes encoding essential Tat pathway components are defective in the integrity of their cell envelope because of the mislocalization of two amidases involved in cell wall metabolism (Ize, B., Stanley, N. R., Buchanan, G., and Palmer, T. (2003) Mol. Microbiol. 48, 1183-1193). To distinguish between genes that are differentially expressed specifically because of the cell envelope defect and those that result from other effects of the tatC deletion, we also analyzed two different transposon mutants of the DeltatatC strain that have their outer membrane integrity restored. Approximately 50% of the genes that were differentially expressed in the tatC mutant are linked to the envelope defect, with the products of many of these genes involved in self-defense or protection mechanisms, including the production of exopolysaccharide. Among the changes that were not explicitly linked to envelope integrity, we characterized a role for the Tat system in iron acquisition and copper homeostasis. Finally, we have demonstrated that overproduction of the Tat substrate SufI saturates the Tat translocon and produces effects on global gene expression that are similar to those resulting from the DeltatatC mutation.


Structural and spectroscopic insight into the metal binding properties of the o-aminophenol-N,N,O-triacetic acid (APTRA) chelator: implications for design of metal indicators.
The o-aminophenol-N,N,O-triacetic acid (APTRA) chelator is employed extensively as a metal-recognition moiety in fluorescent indicators for biological free Mg(2+), as well as in low-affinity indicators for the detection of high levels of cellular Ca(2+). Despite its widespread use in sensor design, the limited metal selectivity of this chelating moiety can lead to binding of competing cations that complicate the fluorescence-based detection of metals of interest in complex samples. Reported herein are the structural characterization of APTRA complexes with various biologically relevant cations, and the thermodynamic analysis of complex formation with Mg(2+), Ca(2+) and Zn(2+). Our results indicate that the low affinity of APTRA for Mg(2+), which makes it a suitable metal-recognition moiety for sensitive analysis of typical millimolar levels of this metal in cells, stems from a much higher enthalpic cost of Mg(2+) binding compared to that of other cations. The results are discussed in the context of indicator design, highlighting the aspects that may aid the future development of fluorescent sensors with enhanced metal selectivity profiles.


New perspectives in equine gastric ulcer syndrome.
Equine gastric ulcer syndrome (EGUS) is common in horses. A history of mild intermitted recurrent colic signs after eating is noted in many horses. Management of horses with abdominal pain caused by gastric ulcers is especially difficult, because non-steroidal anti-inflammatory agents, typically used to control abdominal pain, may exacerbate this condition. Effective pharmacologic agents are available to treat EGUS and eliminate abdominal pain, but more comprehensive measures of environmental and dietary management are needed to manage horses with EGUS and prevent recurrence. This article focuses on the history, clinical signs, diagnosis, and management of horses with abdominal pain associated with gastric ulcers. The primary goal is to provide an understanding of EGUS and to review effective pain management and specific antiulcer treatments and management strategies in horses with EGUS.


Frequency-locked light scattering: real-time Doppler velocimetry with closed-loop feedback control.
Real-time measurement capability of a frequency-modulated filtered light-scattering- (FM FLS) Doppler velocimeter has been demonstrated. Doppler-shifted light from a frequency-modulated Ti:sapphire laser scattered from a supersonic flow is imaged through a potassium vapor cell and is detected by FM spectroscopy. The FM signal is used in closed-loop feedback control of the laser frequency to lock the Doppler-shifted scattered light to the resonance frequency of the filter. The difference between the filter resonance frequency and the laser frequency when the scattered light is frequency locked to the filter resonance is the flow-induced Doppler shift. Changes in flow velocity are tracked by changes in laser frequency, which is subsequently measured to obtain the Doppler shift. The frequency-locking capability of the technique was achieved with use of a simple analog controller. The random Doppler shift measurement errors (2varsigma) were approximately 20 MHz, which correspond to velocity measurement errors for the real-time measurement of less than 3% in a 10-Hz bandwidth.