Increasing the cervical lordosis with chiropractic biophysics seated combined extension-compression and transverse load cervical traction with cervical manipulation: nonrandomized clinical control trial.
Cervical lordosis has been shown to be an important outcome of care; however, few conservative methods of rehabilitating sagittal cervical alignment have been reported. To study whether a seated, retracted, extended, and compressed position would cause tension in the anterior cervical ligament, anterior disk, and muscle structures, and thereby restore cervical lordosis or increase the curvature in patients with loss of the cervical lordosis. Nonrandomized, prospective, clinical control trial. Thirty preselected patients, after diagnostic screening for tolerance to cervical extension with compression, were treated for the first 3 weeks of care using cervical manipulation and a new type of cervical extension-compression traction (vertical weight applied to the subject's forehead in the sitting position with a transverse load at the area of kyphosis). Pretreatment and posttreatment Visual Analogue Scale (VAS) pain ratings were compared along with pretreatment and posttreatment lateral cervical radiographs analyzed with the posterior tangent method for changes in alignment. Results are compared to a control group of 33 subjects receiving no treatment and matched for age, sex, weight, height, and pain. Control subjects reported no change in VAS pain ratings and had no statistical significant change in segmental or global cervical alignment on comparative lateral cervical radiographs (difference in all angle mean values < 1.3 degrees ) repeated an average of 8.5 months later. For the traction group, VAS ratings were 4.1 pretreatment and 1.1 posttreatment. On comparative lateral cervical radiographs repeated after an average of 38 visits over 14.6 weeks, 10 angles and 2 distances showed statistically significant improvements, including anterior head weight bearing (mean improvement of 11 mm), Cobb angle at C2-C7 (mean improvement of -13.6 degrees ), and the angle of intersection of the posterior tangents at C2-C7 (mean improvement of 17.9 degrees ). Twenty-one (70%) of the treatment group subjects were followed for an additional 14 months; improvements in cervical lordosis and anterior weight bearing were maintained. Chiropractic biophysics (CBP) technique's extension-compression 2-way cervical traction combined with spinal manipulation decreased chronic neck pain intensity and improved cervical lordosis in 38 visits over 14.6 weeks, as indicated by increases in segmental and global cervical alignment. Anterior head weight-bearing was reduced by 11 mm; Cobb angles averaged an increase of 13 degrees to 14 degrees; and the angle of intersection of posterior tangents on C2 and C7 averaged 17.9 degrees of improvement.


Letter to the Editor: Diagnostic Criteria in Urological Diseases do not Always Match with Findings by Extended Culture Techniques and Metagenomic Sequencing of 16S rDNA.
Some diseases of the urinary tract are defined by the presence of microorganisms while others are defined by their absence. The underlying idea has always been that urine from healthy subjects is sterile and a negative urine culture has usually been taken as discriminative for an infection to be absent. Several disorders with symptoms that resemble infections are regarded as separate entities based on the exclusion of bacterial growth such as overactive neurogenic bladder and pelvic pain syndromes. During the recent years two paradigmata related to the role of bacteria in urological disease classification have changed completely. Firstly, bacteriuria does not necessarily mean an infection, and secondly, if extended sets of culture media for identification of fastidious and anaerobic bacteria or culture-independent metagenomic sequencing (MGS) is applied, a broad range of even non-culturable bacteria has been detected in the "sterile" bladder urine in healthy individuals. The aim of this editorial is to initiate a discussion to redefine the criteria for urinary tract infections and non-infectious urological disorders with similar symptoms. Clinical studies, in which extended sets of culture media and MGS are integrated, are needed to clarify the pathogenesis of urological disorders where bacteria may play a role. The pure detection of bacteria in the urine does not by itself prove an infectious etiology of a specific disorder. It is important to avoid that results of new technologies lead to unnecessary antibiotic consumption with unwanted collateral damage and adverse events.


Genetic variability in insulin action inhibitor Ikkbeta (IKBKB) does not play a major role in the development of type 2 diabetes.
Recent evidence indicates that IkappaB kinase beta (Ikkbeta) may be a mediator of acquired forms of insulin-resistance. In this study, we examined whether genetic variability at the Ikkbeta locus (IKBKB) contributes to the development of genetic forms of early-onset type 2 diabetes transmitted with an autosomal dominant mode of inheritance. Linkage with four markers flanking the IKBKB gene was evaluated in 32 multigenerational families. Included in the study were 233 diabetic (mean age at Dx = 37 +/- 18) and 152 nondiabetic subjects. The overall LOD scores were negative (-54.9 and -46.2 on the centromeric and telomeric sides, respectively) indicating that variability in IKBKB was not a major determinant of diabetes in these families. Positive values, however, were observed for selected pedigrees. All 17 families for which linkage with the IKBKB locus could not be excluded were screened for sequence differences in the 22 exons and 1.6 kb of the 5' flanking region by dideoxyfingerprinting or direct sequencing. Polymorphisms were identified in the 5' flanking region (-1775del/insC and -1547T > A), exon 11 (c.1083A > G, L361L) and in intron 12 (IVS12+14t > a). However, no mutations segregating with diabetes could be found in these families. Furthermore, all four polymorphisms had similar allele frequencies in the 32 family probands, 171 individuals with common, later-onset type 2 diabetes, and 182 nondiabetic controls. We conclude that sequence differences in the IKBKB gene do not play a major role in either early-onset, autosomal dominant type 2 diabetes, or common forms with a later-onset.


Optics-on-a-chip for ultrafast manipulation of 350-MHz hard x-ray pulses.
Microelectromechanical systems (MEMS) are miniature devices integrated into a vast range of industrial and consumer applications. Optical MEMS are developed for dynamic spatiotemporal control in lightwave manipulation and communication as modulators, switches, multiplexers, spectrometer, etc. However, they have not been shown to function similarly in sub-nm wavelength regimes, namely, with hard x-rays, as high-brilliance pulsed x-rays have proven powerful for addressing challenges in time-domain science, from energy conversion to neurobiological control. While desirable temporal properties of x-ray pulses can be enhanced by optics, conventional x-ray optics are inherently massive in size, hence, never dynamic. We demonstrate highly ultrafast x-ray optics-on-a-chip based on MEMS capable of modulating hard x-ray pulses exceeding 350 MHz, 103× higher than any other mechanical modulator, with a pulse purity >106 without compromising the spectral brilliance. Moreover, the timing characteristics of the devices can be tuned on-the-fly to deliver optimal pulse properties to create a host of dynamic x-ray instruments and applications, impossible with traditional optics of 109× bulkier and more massive. The advent of the ultrafast optics-on-a-chip heralds a new paradigm of x-ray photonics, time-domain science, and accelerator diagnostics, especially at not only the future-generation light sources that offer coherent and high-frequency pulses but also lab-based facilities that normally do not offer timing structures.


[Preparative subfractionation of high-density lipoproteins in dysalphalipoproteinemias].
A new modified procedure is described for preparative production of three subfractions of high density lipoproteins (HDL) by means of salt density gradient developed in ultracentrifugation. Concentrations of cholesterol, phospholipids and apoprotein A-1 in both subfractions HDL2b and HDL2 alpha were found to alter simultaneously with the level of HDL cholesterol in blood plasma of the patients with low, normal and increased content of HDL cholesterol (10 men in each group), thus suggesting the alteration of the particles amount in both these subfractions. In hyperalphacholesterolemia the level of all the components studied was unaltered as compared with normal state, and in hypoalphacholesterolemia only cholesterol containing in HDL3 was decreased due to diminished loading of the particles with cholesterol, as calculated per a unit of apoprotein A-1. The data obtained suggest that alterations in amount of both HDL2b and HDL2a particles depended on intensity of lipid binding, especially of cholesterol, to particles of the HDL3 subfraction, which plays an important role in various physiological and pathological states.


RCRdiff: A fully integrated Bayesian method for differential expression analysis using raw NanoString nCounter data.
The medium-throughput mRNA abundance platform NanoString nCounter has gained great popularity in the past decade, due to its high sensitivity and technical reproducibility as well as remarkable applicability to ubiquitous formalin fixed paraffin embedded (FFPE) tissue samples. Based on RCRnorm developed for normalizing NanoString nCounter data and Bayesian LASSO for variable selection, we propose a fully integrated Bayesian method, called RCRdiff, to detect differentially expressed (DE) genes between different groups of tissue samples (eg, normal and cancer). Unlike existing methods that often require normalization performed beforehand, RCRdiff directly handles raw read counts and jointly models the behaviors of different types of internal controls along with DE and non-DE gene patterns. Doing so would avoid efficiency loss caused by ignoring estimation uncertainty from the normalization step in a sequential approach and thus can offer more reliable statistical inference. We also propose clustering-based strategies for DE gene selection, which do not require any external dataset and are free of any arbitrary cutoff. Empirical evidence of the attractiveness of RCRdiff is demonstrated via extensive simulation and data examples.


Nrf1 and Nrf2 play distinct roles in activation of antioxidant response element-dependent genes.
Nrf1 is a member of the vertebrate Cap'n'Collar (CNC) transcription factor family that commonly contains a unique basic-leucine zipper domain. Among CNC family members, Nrf2 is known to regulate a battery of antioxidant and xenobiotic-metabolizing enzyme genes through the antioxidant response element (ARE). Although Nrf1 has also been shown to bind the ARE, it is unclear whether it plays a distinct role from Nrf2 in regulating genes with this element. To address this issue in vivo, we generated mice bearing a hepatocyte-specific disruption of the Nrf1 gene. AlthoughNrf2 knock-out mice did not exhibit liver damage when they were maintained in an unstressed condition, hepatocyte-specific deletion of Nrf1 caused liver damage resembling the human disease non-alcoholic steatohepatitis. Gene expression analysis revealed that the disruption of Nrf1 causes stress that activates a number of ARE-driven genes in an Nrf2-dependent manner, indicating that Nrf2 cannot compensate completely for loss of Nrf1 function in the liver. In contrast, expression of metallothionein-1 and -2 (MT1 and MT2) genes, each of which harbors at least one ARE in its regulatory region, was decreased in the Nrf1-null mutant mice. Whereas Nrf1 and Nrf2 bound the MT1 ARE with comparable affinity, Nrf1 preferentially activated the reporter gene expression through the MT1 ARE. This study has, thus, identified the first ARE-dependent gene that relies exclusively on Nrf1, suggesting that it plays a distinct functional role in regulating ARE-driven genes.


A total laparoscopic approach reduces the infertility rate after ileal pouch-anal anastomosis: a 2-center study.
To assess the infertility rate after laparoscopic ileal pouch-anal anastomosis (IPAA). Total proctocolectomy with IPAA is known to be associated with postoperative infertility in open surgery, which may be caused by pelvic adhesions affecting the fallopian tubes. However, fertility after laparoscopic IPAA has never been assessed. All patients who underwent a total laparoscopic IPAA between 2000 and 2011 and were aged 45 years or less at the time of operation and 18 years or more at the time of data collection were included. The patients answered a fertility questionnaire by telephone. All demographic and perioperative data were prospectively collected. The results were compared with those of controls undergoing laparoscopic appendectomy. Sixty-three patients were included. The mean age at the time of surgery was 31 ± 9 years (range 14-44). IPAA was performed for ulcerative colitis in 73% of the cases and familial adenomatous polyposis in 17%. The mean follow-up after IPAA was 68 ± 33 months (range 6-136). Fifty-six patients answered the questionnaire (89%). Half of them already had a child before IPAA. Fifteen patients attempted pregnancy after IPAA, of which 11 (73%) were able to conceive, resulting in 10 ongoing pregnancies and 1 miscarriage. The global infertility rate was 27%. There was no difference in fertility over time compared with the 14 controls who attempted pregnancy during the same period (90% vs 86% at 36 months, P = 0.397). The infertility rate appears to be lower after laparoscopic IPAA than after open surgery.


Surgical management for hydatid disease.
Hydatid disease is caused by the parasite Echinococcus granulosus. The liver and the lungs are common sites. When a cystic lesion is seen on CT scan, diagnosis is made based on the patient having lived in an endemic area. Serologic tests are used for differential diagnosis. Medical treatment is centered on albendazole. Surgery is recommended either by open or endoscopic technique depending on the characteristics of the cysts and the patient. Complications of surgery are rare except for prolonged air leaks. Mortality occurs when the cyst is located in the central nervous system or occludes major vessels.


Peer rejection, affiliation with deviant peers, delinquency, and risky sexual behavior.
Risky sexual behavior poses significant health risks by increasing sexually transmitted infections and unintended pregnancies. Previous research has documented many factors related to risky sexual behavior. This study adds to the literature by proposing a prospective, developmental model of peer factors related to risky sexual behavior. Developmental pathways to risky sexual behavior were examined in a sample of 517 individuals (51% female; 82% European American, 16% African American, 2% other) followed from age 5-27. Structural equation models examined direct and indirect effects of peer rejection (assessed via peer nominations at ages 5, 6, 7, and 8), affiliation with deviant peers (assessed via self-report at ages 11 and 12), and delinquency (assessed via maternal report at ages 10 and 16) on risky sexual behavior (assessed via self-report at age 27). More peer rejection during childhood, affiliation with deviant peers during pre- adolescence, and delinquency in childhood and adolescence predicted more risky sexual behavior through age 27, although delinquency at age 16 was the only risk factor that had a significant direct effect on risky sexual behavior through age 27 above and beyond the other risk factors. Peer rejection was related to subsequent risk factors for girls but not boys. Peer risk factors as early as age 5 shape developmental pathways through childhood and adolescence and have implications for risky sexual behavior into adulthood.


[Integrationism versus nonconformism. Two slogans and two programs].
The problem concerning the alternative between integration and segregation can be understood in a more adequate manner as the alternative between integrationism and nonconformism. Deafness serves as a paradigmatic model which helps to understand that, in order to warrant a healthy emotional development, the child and adolescent need to enjoy from both opportunities: raising of specially characteristic features as well as promoting common social patterns. --Fully integrated education is therefore as insufficient as is a just segregated one.


Importance of cardiac rehabilitation in rural and remote areas of Australia.
To assess implementation of in-patient cardiac rehabilitation (Phase-1-cardiac rehabilitation), impact on people in rural and remote areas of Australia and potential methods for addressing identified weaknesses. Exploratory case study methodology using qualitative and quantitative methods. Qualitative data collection via semi-structured interviews, using thematic analysis, augmented by quantitative data collection via a medical record audit. Four regional hospitals (2 Queensland Health and 2 private) providing tertiary health care. (a) Hospital in-patients with heart disease ≥18 years. (b) Staff responsible for their care. Implementation of Phase-1-cardiac rehabilitation in tertiary hosptials in North Queensland and the impact on in-patients discharge planning and post discharge care. Recommentations and implications for practice are proposed to address deficits. Phase-1-cardiac rehabilitation implementation rates, in-patient understanding and multidisciplinary team involvement were low. The highest rates of Phase-1-cardiac rehabilitation were for in-patients with a length of stay three days or more in cardiac units with cardiac educators. Rates were lower in cardiac units with no cardiac educators, and lowest for in-patients in all areas of all hospitals with length of stay of two days or less days. Low Phase-1-cardiac rehabilitation implementation rates resulted in poor in-patient understanding about their disease, treatment and post-discharge care. Further, medical discharge summaries rarely mentioned cardiac rehabilitation/secondary prevention or risk factor management resulting in a lack of information for health care providers on cardiac rehabilitation and holistic health care. Implementation of Phase-1-cardiac rehabilitation in regional hospitals in this study fell short of recommended best practice, resulting in patients' poor preparation for discharge, and insufficient information on holistic care for health care providers in rural and remote areas. These factors potentially impact on holistic care for people returning home following treatment for heart disease.


A pathophysiologic, gastroenterologic, and radiologic approach to the management of gastric varices.
Gastric varices (GV) occur in 20% of patients with portal hypertension either in isolation or in combination with esophageal varices (EV). There is no consensus for optimum treatment of GV and because they comprise an inhomogeneous entity, accurate classification is vital to determine the appropriate management. Gastroesophageal varices (GOV) are classified as GOV1 (EV extending down to cardia or lesser curve) or GOV2 (esophageal and fundal varices). Isolated gastric varices (IGV) may be located in the fundus (IGV1) or elsewhere in the stomach (IGV2). GV possibly bleed less frequently than EV, but GV bleeding is typically difficult to control, associated with a high risk for rebleeding, and high mortality. Fundal varices, large GV (>5 mm), presence of a red spot, and Child's C liver status are associated with a high risk for bleeding. GOV1 have a much lower risk for bleeding. A portosystemic pressure gradient of > or =12 mm Hg is not necessary for GV bleeding, probably related to the high frequency of spontaneous gastrorenal shunts in these patients. GOV1 should be treated as for EV. First-line treatment of bleeding fundal varices is endoscopic variceal obturation. TIPS is currently second-line acute treatment and is used for prevention of rebleeding. The role of some newer interventional radiologic techniques requires further appraisal. This review describes the pathophysiology, diagnosis, natural history, endoscopic, and interventional radiologic treatment options for GV.


Rationally Designed Vaccines Targeting the V2 Region of HIV-1 gp120 Induce a Focused, Cross-Clade-Reactive, Biologically Functional Antibody Response.
Strong antibody (Ab) responses against V1V2 epitopes of the human immunodeficiency virus type 1 (HIV-1) gp120 envelope (Env) correlated with reduced infection rates in studies of HIV, simian-human immunodeficiency virus (SHIV), and simian immunodeficiency virus (SIV). In order to focus the Ab response on V1V2, we used six V1V2 sequences and nine scaffold proteins to construct immunogens which were tested using various immunization regimens for their ability to induce cross-reactive and biologically active V2 Abs in rabbits. A prime/boost immunization strategy was employed using gp120 DNA and various V1V2-scaffold proteins. The rabbit polyclonal Ab responses (i) were successfully focused on the V1V2 region, with weak or only transient responses to other Env epitopes, (ii) displayed broad cross-reactive binding activity with gp120s and the V1V2 regions of diverse strains from clades B, C, and E, (iii) included V2 Abs with specificities similar to those found in HIV-infected individuals, and (iv) remained detectable ≥1 year after the last boosting dose. Importantly, sera from rabbits receiving V1V2-scaffold immunogens displayed Ab-dependent cellular phagocytosis whereas sera from rabbits receiving only gp120 did not. The results represent the first fully successful example of reverse vaccinology in the HIV vaccine field with rationally designed epitope scaffold immunogens inducing Abs that recapitulate the epitope specificity and biologic activity of the human monoclonal Abs from which the immunogens were designed. Moreover, this is the first immunogenicity study using epitope-targeting, rationally designed vaccine constructs that induced an Fc-mediated activity associated with protection from infection with HIV, SIV, and SHIV. Novel immunogens were designed to focus the antibody response of rabbits on the V1V2 epitopes of HIV-1 gp120 since such antibodies were associated with reduced infection rates of HIV, SIV, and SHIV. The vaccine-induced antibodies were broadly cross-reactive with the V1V2 regions of HIV subtypes B, C and E and, importantly, facilitated Fc-mediated phagocytosis, an activity not induced upon immunization of rabbits with gp120. This is the first immunogenicity study of vaccine constructs that focuses the antibody response on V1V2 and induces V2-specific antibodies with the ability to mediate phagocytosis, an activity that has been associated with protection from infection with HIV, SIV, and SHIV.


Human proximity seeking in family pigs and dogs.
Family dogs (Canis familiaris) seek human contact from an early age, can discriminate and prefer their caregivers over other humans. To investigate if being kept as a family animal is sufficient to induce similar early human proximity-seeking in another domestic mammal, here we directly compared such behaviours in dogs and similarly raised domestic pigs (Sus scrofa domesticus). We used a preference test to measure proximity-seeking behaviours of 4-month-old family pigs and dogs in the presence of their caregiver and either a stranger or a familiar object, in a novel environment. We found that both pigs and dogs preferred their caregivers over the familiar object but not over the stranger. However, when the caregiver and the stranger were present, only dogs showed an overall preference for human proximity, and pigs spent more time away from both humans. These results suggest that both dogs and pigs seek the proximity of their caregiver, but there is a difference in how each species generalizes their experience to other humans. Species-specific predispositions, including that dogs have a longer socialization period and that humans are more salient as a social stimulus for them, may be important for the early development of an overall preference for humans.


Adult hematopoietic stem cells require NKAP for maintenance and survival.
Steady-state hematopoiesis is sustained through differentiation balanced with proliferation and self-renewal of hematopoietic stem cells (HSCs). Disruption of this balance can lead to hematopoietic failure, as hematopoietic differentiation without self-renewal leads to loss of the HSC pool. We find that conditional knockout mice that delete the transcriptional repressor NKAP in HSCs and all hematopoietic lineages during embryonic development exhibit perinatal lethality and abrogation of hematopoiesis as demonstrated by multilineage defects in lymphocyte, granulocyte, erythrocyte and megakaryocyte development. Inducible deletion of NKAP in adult mice leads to lethality within 2 weeks, at which point hematopoiesis in the bone marrow has halted and HSCs have disappeared. This hematopoietic failure and lethality is cell intrinsic, as radiation chimeras reconstituted with inducible Mx1-cre NKAP conditional knockout bone marrow also succumb with a similar time course. Even in the context of a completely normal bone marrow environment using mixed radiation chimeras, NKAP deletion results in HSC failure. NKAP deletion leads to decreased proliferation and increased apoptosis of HSCs, which is likely due to increased expression of the cyclin-dependent kinase inhibitors p21Cip1/Waf1 and p19Ink4d. These data establish NKAP as one of a very small number of transcriptional regulators that is absolutely required for adult HSC maintenance and survival.


COVID-19: An Update on Pathogenesis and Treatment.
In 2019, a new virus -SARS-COV2 possibly emerged in China, which infected many people affecting mainly the respiratory system. SARS-COV2 gets transmitted by inhalation of droplets from the infected persons. Symptoms start to appear after the incubation period of the virus which ranges from 2 to 14 days. In most people, symptoms are usually mild such as fever, sore throat, cough, chest tightness and fatigue. In other people, the disease might progress into severe pneumonia leading to several fatal consequences. Treatment is usually supportive and the role of antiviral is not established yet. Home isolation for mild cases is important for the prevention of the transmission of infection. Although the rate of transmission of this virus is faster than other viruses from the family, such as MERS-CoV, it has a lower fatality rate. The main difference in the genome structure of this family, which makes it distinguishable from other viruses is its use of (+) ssRNA as genetic material, which comprise 5' cap located at one end and 3' polyadenylation tract at the other end. During infection of an exposed host cell, viral-encoded protease cleaves the polyprotein that results from translation of 5' open reading frame (ORF) of the genome, culminating in the release of multiple nonstructural proteins such as helicase (Hel), adenosine triphosphate (ATPase) and RNA-dependent RNA polymerase (Rep). These proteins are responsible for the replication process in addition to the syntheses of the sub genomic mRNA used as transcription template strand. In this review article, we discussed the transmission pathways, genetic sequence and current treatment approach of COVID-19.


General Washington and the patriot soldiers. They won a war with little food. A Bicentennial study.
The troops in the first action of the American Revolution in Massachusetts were well provisioned through the efforts of Joseph Trumbull of Connecticut, whom the Second Continental Congress appointed Commissary General, at General Washingtion's request. The ration, as ordered, would have provided-by today's standards-more calories, twice as much protein, and adequate minerals and vitamins, except possible vitamin A and ascorbic acid. Unfortunately, all items in the ration were seldom available to the soldiers. Each man was issued his ration and was responsible for cooking it. Trumbull held his post for two years, with little support from the Congress and no departmental organization. When the Army moved to New York, difficulties in provisioning the troops on the march began-and continued until 1781. Following the retreat to Morristown, New Jersey, in 1776, flour and beef, but little else, were available during the winter and into the following summer. Washington pleaded with the Congress for help with food supplies, but to no avail. Finally, Trumbull resigned. Thereafter, followed a series of four reorganizations of the commissariat by the Congress-each disastrously conceived-which brought no relief to the ill-fed soldiers, whose physical condition progressively worsened. Finally, a fifth reorganization in 1781 put into operation a contract plan of procuring food; no longer was it necessary for Washington to order impressment of food from farmers or for the troops to live off the land. The Continental Army "starved, not because the country could not furnish the food, but becasue the people were unwilling to endure taxation and Congress did not understand the importance of administrative centralization; ...the greater part of the hardships" were avoidable. Yes, General Washington and the patriot soldiers won the war with little food.


[The molecular biology of epithelial ovarian cancer].
Epithelial ovarian cancer frequently presents at an advanced stage where the cornerstone of management remains surgery and platinum-based chemotherapy. Unfortunately, despite sometimes dramatic initial responses, advanced ovarian cancer almost invariably relapses. Little progress has been made in the identification of effective targeted-therapies for ovarian cancer. The majority of clinical trials investigating novel agents have been negative and the only approved targeted-therapy is bevacizumab, for which reliable predictive biomarkers still elude us. Ovarian cancer is treated as a uniform disease. Yet, biological studies have highlighted the heterogeneity of this malignancy with marked differences in histology, oncogenesis, prognosis, chemo-responsiveness, and molecular profile. Recent high throughput molecular analyses have identified a huge number of genomic/phenotypic alterations. Broadly speaking, high grade serous carcinomas (type II) display significant genomic instability and numerous amplifications and losses; low grade (type I) tumors are genomically stable but display frequent mutations. Importantly, many of these genomic alterations relate to known oncogenes for which targeted-therapies are available or in development. There is today a real potential for personalized medicine in ovarian cancer. We will review the current literature regarding the molecular characterization of epithelial ovarian cancer and discuss the biological rationale for a number of targeted strategies. In order to translate these biological advances into meaningful clinical improvements for our patients, it is imperative to incorporate translational research in ovarian cancer trials, a number of strategies will be proposed such as the acquisition of quality tumor samples, including sequential pre- and post-treatment biopsies, the potential of liquid biopsies, and novel trial designs more adapted to the molecular era of ovarian cancer research.


The Di2/pet Variant in the PETALOSA Gene Underlies a Major Heat Requirement-Related QTL for Blooming Date in Peach [Prunus persica (L.) Batsch].
Environmental adaptation of deciduous fruit trees largely depends on their ability to synchronize growth and development with seasonal climate change. Winter dormancy of flower buds is a key process to prevent frost damage and ensure reproductive success. Temperature is a crucial environmental stimulus largely influencing the timing of flowering, only occurring after fulfillment of certain temperature requirements. Nevertheless, genetic variation affecting chilling or heat-dependent dormancy release still remains largely unknown. In this study, a major QTL able to delay blooming date in peach by increasing heat requirement was finely mapped in three segregating progenies, revealing a strict association with a genetic variant (petDEL) in a PETALOSA gene, previously shown to also affect flower morphology. Analysis of segregating genome-edited tobacco plants provided further evidence of the potential ability of PET variations to delay flowering time. Potential applications of the petDEL variant for improving phenological traits in peach are discussed.


Participatory design methods for the development of a clinical telehealth service for neonatal homecare.
Neonatal homecare delivered during home visits by neonatal nurses is a common method for supporting families of preterm infants following discharge. Telehealth has been introduced for the provision of neonatal homecare, resulting in positive feedback from parents of preterm infants. While the benefits are beginning to be realised, widespread uptake of telehealth has been limited due to a range of logistical challenges. Understanding user requirements is important when planning and developing a clinical telehealth service. We therefore used participatory design to develop a clinical telehealth service for neonatal homecare. The study adopted a participatory design approach to engage users in the development and design of a new telehealth service. Participatory design embraces qualitative research methods. Creative and technical workshops were conducted as part of the study. Tests of the telehealth service were conducted in the neonatal unit. Participants in this study were former and current parents of preterm infants eligible for neonatal homecare, and clinical staff (medical and nursing) from the neonatal unit. Preterm infants accompanied their parents. Based on the results obtained during the workshops and subsequent testing, we developed an application (app), which was integrated into the medical record at the neonatal unit. The app was used to initiate videoconferences and chat messages between the family at home and the neonatal unit, and to share information regarding infant growth and well-being. Results obtained from the workshops and testing demonstrated the importance of involving users when developing new telehealth applications. The workshops helped identify the challenges associated with delivery of the service, and helped instruct the design of a new telehealth service for neonatal homecare based on the needs of parents and clinical staff.


Clinical Pharmacology of Janus Kinase Inhibitors in Inflammatory Bowel Disease.
Inflammatory bowel disease, including ulcerative colitis and Crohn's disease, are chronic inflammatory disorders of the gastrointestinal tract which are characterised, in part, by an imbalance in the production of several pro- and anti-inflammatory cytokines. Although various agents are effective for inducing and maintaining remission, approximately 20% of patients are treatment-refractory and require surgery. Parenterally administered monoclonal antibody-based biologics are associated with adverse effects resulting in treatment discontinuation and/or immunogenicity, leading to loss of response to therapy. Approximately 50% of patients who initially respond to treatment with tumour necrosis factor antagonists lose response to therapy within the 1st year of treatment. Incidence of immunogenicity tends to decrease over time, but once present can persist for years, even after treatment discontinuation. Nonimmunogenic oral small molecule therapies, including Janus kinase inhibitors, are currently being developed and have demonstrated efficacy in early phase clinical trials, which has already led to regulatory approval of tofacitinib for the treatment of patients with moderate-to-severe ulcerative colitis. Differentiation of T cells into T helper cells, which are mediators of the inflammatory response in inflammatory bowel disease, is mediated by the Janus kinase signal transducer and activator of the transcription signalling pathway. Absorption and distribution of Janus kinase inhibitors occurs at the site of action in the gastrointestinal tract, and newer compounds are being developed with limited systemic absorption, potentially reducing the risk of adverse effects. The current review describes the clinical pharmacology of approved Janus kinase inhibitors, as well as those in clinical development for the treatment of inflammatory bowel disease.


[Chemical constituents analysis of Mgrtol standardized by GC-FTIR].
The chemical constituents of the Mgrtol standardized were analyzed by capillary gas chromatography-Fourier transform infrared spectroscopy (GC-FTIR). The relative content of each component was determined by area nomalization method. Twelve peaks were separated and were all identified by FTIR through search method. The results showed that the Mgrtol standardized mainly consists of 3 kinds of single terpenes, including 1,8-cineole, D-limonen and alpha-pinene, which account for 14.70%, 36.83% and 46.35% respectively, totaling to 97.88% of the entire compositions of the Mgrtol standardized. The results are the same as those analyzed by GC-MS. Comparing with GC-MS, the isomers of organic compounds separated by gas chromatography could be determined by GC-FTIR. So this method has wider application value. The results also provided a scientific proof for the further development of the plant.


Disruption of zinc homeostasis and the pathogenesis of senile dementia.
Zinc (Zn) is an essential trace element that is abundantly present in the brain. Although Zn plays crucial roles in learning and memory, numerous studies have indicated that the disruption of Zn homeostasis, namely both depletion and excess Zn, causes severe damage to neurons and is linked with various neurodegenerative diseases including Alzheimer's disease and vascular dementia. Here, we review the current understanding of the role of Zn in the pathogenesis of these neurodegenerative diseases. Based on our findings and other numerous studies, Zn acts as a contributor to Alzheimer's disease in the oligomerization, and as a protector in the neurotoxicity of Alzheimer's β-amyloid protein. Furthermore, Zn plays a central role in ischemia-induced neuronal death and the pathogenesis of vascular dementia. Involvement of Ca(2+) dyshomeostasis and endoplasmic reticulum (ER) stress in the mechanism of Zn-induced neurotoxicity are suggested. We also discuss the possible role of carnosine (β-alanyl histidine), a dipeptide that is present in the brain, as a protective substance for neuronal injury.


A Single-Run HPLC-MS Multiplex Assay for Therapeutic Drug Monitoring of Relevant First- and Second-Line Antibiotics in the Treatment of Drug-Resistant Tuberculosis.
The treatment of drug-resistant Mycobacterium tuberculosis relies on complex antibiotic therapy. Inadequate antibiotic exposure can lead to treatment failure, acquired drug resistance, and an increased risk of adverse events. Therapeutic drug monitoring (TDM) can be used to optimize the antibiotic exposure. Therefore, we aimed to develop a single-run multiplex assay using high-performance liquid chromatography-mass spectrometry (HPLC-MS) for TDM of patients with multidrug-resistant, pre-extensively drug-resistant and extensively drug-resistant tuberculosis. A target profile for sufficient performance, based on the intended clinical application, was established and the assay was developed accordingly. Antibiotics were analyzed on a zwitterionic hydrophilic interaction liquid chromatography column and a triple quadrupole mass spectrometer using stable isotope-labeled internal standards. The assay was sufficiently sensitive to monitor drug concentrations over five half-lives for rifampicin, rifabutin, levofloxacin, moxifloxacin, bedaquiline, linezolid, clofazimine, terizidone/cycloserine, ethambutol, delamanid, pyrazinamide, meropenem, prothionamide, and para-amino salicylic acid (PAS). Accuracy and precision were sufficient to support clinical decision making (≤±15% in clinical samples and ±20-25% in spiked samples, with 80% of future measured concentrations predicted to fall within ±40% of nominal concentrations). The method was applied in the TDM of two patients with complex drug-resistant tuberculosis. All relevant antibiotics from their regimens could be quantified and high-dose therapy was initiated, followed by microbiological conversion. In conclusion, we developed a multiplex assay that enables TDM of the relevant first- and second-line anti-tuberculosis medicines in a single run and was able to show its applicability in TDM of two drug-resistant tuberculosis patients.


Service delivery through public health care system to control sexually transmitted infections in Himachal pradesh.
The National AIDS Control Organization has designed multiple synergistic interventions to identify and control curable sexually transmitted infections (STIs). To assess the impact of services offered at designated STI clinics in the state of Himachal Pradesh, India and the profile of the attending clients. This was a two-year prospective study, conducted from April 2011 to March 2013. Training on delivering STI/RTI services was imparted to the staff of 16 designated STI clinics including recording of data. The staff in each STI clinic comprises of one doctor, one counselor, one nurse, and one laboratory technician. The clients attending these designated clinics were offered counseling, syndromic case management (SCM), and diagnostic services wherever possible. Monthly data of STI clinic attendees was collected, compiled, and analyzed. A total of 65,760 clinic visits were reported, of which 32,385 (49%) visits were for index STI/RTI complaint(s). The ratio of male to female attendees was 1:2. The commonest age group accessing the STI clinics was 25-44 years (n = 38,966; 59.3%). According to SCM, 52.9% clients were managed. The commonest presenting syndrome was urethral discharge (n = 4,500; 41%) in males, and vaginal discharge (n = 13,305; 56%) in females. Genital ulcer disease was treated in 2099 cases. Laboratory tests were performed only in 6466 patients, and 39,597 antenatal mothers were screened for syphilis. Counseling services were provided to 51,298 (f = 34,804; 68%: m = 16,494; 32%) clients and of these, 48% (n = 25,056) of the clients were referred to integrated counseling and testing centers. Forty-three clients (m = 24: f = 19) were detected positive for HIV infection. Uniform and standardized services delivered to clients attending public health clinics can gather reliable data to monitor trends of STI infection.


Surface area-to-volume ratio, not cellular viscoelasticity, is the major determinant of red blood cell traversal through small channels.
The remarkable deformability of red blood cells (RBCs) depends on the viscoelasticity of the plasma membrane and cell contents and the surface area to volume (SA:V) ratio; however, it remains unclear which of these factors is the key determinant for passage through small capillaries. We used a microfluidic device to examine the traversal of normal, stiffened, swollen, parasitised and immature RBCs. We show that dramatic stiffening of RBCs had no measurable effect on their ability to traverse small channels. By contrast, a moderate decrease in the SA:V ratio had a marked effect on the equivalent cylinder diameter that is traversable by RBCs of similar cellular viscoelasticity. We developed a finite element model that provides a coherent rationale for the experimental observations, based on the nonlinear mechanical behaviour of the RBC membrane skeleton. We conclude that the SA:V ratio should be given more prominence in studies of RBC pathologies.


Protein A-immobilized microporous polyhydroxyethylmethacrylate affinity membranes for selective sorption of human-immunoglobulin-G from human plasma.
Microporous membranes made of poly(2-hydroxyethylmethacrylate) [poly(HEMA)] carrying protein A were used for selective sorption of human-IgG from human plasma. Poly(HEMA) membranes were prepared by a photo-polymerization technique, and activated by cyanogen bromide (CNBr) in an alkaline medium (pH 11.5). Bioligand protein A was then immobilized by covalent binding onto these CNBr-activated membranes. The amount of immobilized protein A was controlled by changing pH and the initial concentrations of CNBr and protein A. The non-specific adsorption of protein A on the plain poly(HEMA) membranes was 2.9 microg cm(-2). Maximum protein A immobilization was observed at pH 9.5. Up to 186 microg cm(-2) was immobilized on the CNBr-activated poly(HEMA) membranes. The maximum adsorption of human-IgG on the protein A-immobilized poly(HEMA) membranes was observed at pH 8.0. The non-specific adsorption of human-IgG onto the plain poly(HEMA) membranes was low (about 4.4 microg cm(-2)). Higher human-IgG adsorption values (up to 394 microg cm(-2)) were obtained in which the protein A-immobilized poly(HEMA) membranes were used. Much higher amounts of human-IgG (up to 489 microg cm(-2)) were adsorbed from human plasma. Up to 91% of the adsorbed human-IgG was desorbed by using 0.1 M aminoacetic acid as elution agent. The adsorption-desorption cycle was repeated ten times using the same polymeric membranes. There was no remarkable reduction in the adsorption capacity of the protein A-immobilized poly(HEMA) membranes.


Assessing the Effect of a Glaucoma Surgical Curriculum in Resident Physicians.
Subspecialty surgical training is an important part of resident education. We changed the glaucoma rotation in which postgraduate year 4 residents worked with multiple attending physicians with varying teaching styles to a structured surgical curriculum led by 2 dedicated preceptors, and we evaluated the effect on residents' surgical performance prospectively. A curriculum consisting of preoperative training, intraoperative teaching, postoperative feedback, and repetition was implemented for postgraduate year 4 residents between July 2, 2012, and June 30, 2014. In a class of 8 residents per year, the mean (SD) glaucoma surgical volume increased from 8.9 (0.8) cases in the prior year to 13.6 (2.5) in 2013 (mean difference, 4.8 cases; 95% CI, 2.4-7.1; P = .001) and 14.8 (4.2) in 2014 (mean difference, 5.9 cases; 95% CI, 2.1-9.6; P = .007). A self-assessment survey showed improvement in suturing (scores for each section range from 1 [worst] to 5 [best]; mean rating, 3.9 in the prior year vs 4.4 in 2013 [P = .04] and 4.5 in 2014 [P = .02]). A validated survey assessing overall surgical competency revealed improvement in handling adverse events (mean rating, 4.1 in the prior year vs 5.0 for both 2013 and 2014; both P < .001). Despite the small sample size and nonrandomized study design, these data suggest that a structured surgical curriculum has advantages in teaching subspecialty surgery and might be considered by other ophthalmology training programs.


Evidence for the presence of xenopsin-related peptide(s) in the gastric mucosa of mammals.
Using immunohistochemistry and radioimmunoassay, substance(s) related to the amphibian octapeptide xenopsin (XP) were demonstrated in the gastric mucosa of humans and dogs. Immunohistochemistry localized XP-immunoreactive epithelial cells in the gastric antral mucosa. The reaction was abolished by preabsorption of the antiserum with XP but not by neurotensin or other peptides. Immunoreactive XP (iXP) was found by radioimmunoassay in extracts of both the antrum and body of the stomach prepared with acid/acetone or acetic acid. A study of its distribution in the dog indicated that the level of iXP was highest in the stomach, lower in the pancreas and duodenum, and not measurable in the jejunoileum and colon. Gel chromatography on Sephadex G-25 indicated the presence of at least two forms of iXP, one larger and the other about the same size as XP. Reverse-phase high pressure liquid chromatography on mu-Bondapak C-18 yielded several peaks of iXP, one of which eluted at the position of synthetic XP. The results of immunochemical analyses using four different antisera towards XP were consistent with structures for canine iXPs that were closely related to XP only in their C-terminal regions. These results suggest that mammalian counterparts to amphibian XP reside within endocrine cells of the gastric mucosa. It seems possible that these peptides function as gastrointestinal signals.


A retrieval analysis perspective on revision for infection.
Retrieval analysis has long served the orthopaedic community as a tool for understanding implant failure modes; however, what retrieval studies can reveal about the nature of prosthetic joint infection (PJI) remains unknown. We hypothesize that records from a comprehensive joint retrieval program should corroborate clinically-reported temporal characteristics of prosthesis-related infection. We examined 2527 records documenting a decade of explanted hip and knee components to quantify the following: (1) the relative contribution of infection to revision arthroplasty; (2) the effects of joint type, revision status, and reason for retrieval on indwelling time; and (3) whether the temporal distribution of infected explants reflects clinical experience. In this series, 20% (507/2527) of explants were performed for infection, with PJI being more commonly implicated in the retrieval of revision implants than of primaries. Infected prostheses were explanted 23.2 months sooner on average than those retrieved for other causes. Within the subset of infected devices, revision components were explanted 11.2 months sooner than primaries, with no appreciable difference observed between hips and knees. Retrieval-based temporal distributions were most similar to PJI studies with endpoint enrollment or long duration follow-up, suggesting a later average onset of infection than reported in comparable clinical studies with short (<10-year) follow-up. Infection represents a major cause of revision arthroplasty, and is associated with shorter indwelling times in revision components than in primaries. Studies with less than 10 years of follow-up are likely to under-report late PJI.


The role of the perfusionist.
During open heart procedures, the heart is stopped. The surgeon needs a blood-free, motionless work area. The heart-lung bypass machine provides circulation for the rest of the body during surgery. The perfusionist, as an active member of the surgical team, operates the heart-lung bypass machine. Historical development of the perfusionist's role and collaboration with other members of the healthcare team are discussed.


First imaging results of an intraindividual comparison of (11)C-acetate and (18)F-fluorocholine PET/CT in patients with prostate cancer at early biochemical first or second relapse after prostatectomy or radiotherapy.
(18)F-Fluorocholine (FCH) and (11)C-acetate (ACE) PET are widely used for detection of recurrent prostate cancer (PC). We present the first results of a comparative, prospective PET/CT study of both tracers evaluated in the same patients presenting with recurrence and low PSA to compare the diagnostic information provided by the two tracers. The study group comprised 23 patients studied for a rising PSA level after radical prostatectomy (RP, 7 patients, PSA ≤ 3 ng/ml), curative radiotherapy (RT, 7 patients, PSA ≤ 5 ng/ml) or RP and salvage RT (9 patients, PSA ≤ 5 ng/ml). Both FCH and ACE PET/CT scans were performed in a random sequence a median of 4 days (range 0 to 11 days) apart. FCH PET/CT was started at injection (307 ± 16 MBq) with a 10-min dynamic acquisition of the prostate bed, followed by a whole-body PET scan and late (45 min) imaging of the pelvis. ACE PET/CT was performed as a double whole-body PET scan starting 5 and 22 min after injection (994 ± 72 MBq), and a late view (45 min) of the prostate bed. PET/CT scans were blindly reviewed by two independent pairs of two experienced nuclear medicine physicians, discordant subgroup results being discussed to reach a consensus for positive, negative end equivocal results. PET results were concordant in 88 out of 92 local, regional and distant findings (Cohen's kappa 0.929). In particular, results were concordant in all patients concerning local status, bone metastases and distant findings. Lymph-node results were concordant in 19 patients and different in 4 patients. On a per-patient basis results were concordant in 22 of 23 patients (14 positive, 5 negative and 3 equivocal). In only one patient was ACE PET/CT positive for nodal metastases while FCH PET/CT was overall negative; interestingly, the ACE-positive and FCH-negative lymph nodes became positive in a second FCH PET/CT scan performed a few months later. Overall, ACE and FCH PET/CT showed excellent concordance, on both a per-lesion and a per-patient basis, suggesting that both tracers perform equally for recurrent prostate cancer staging.


Trichosporin-B-III, an alpha-aminoisobutyric acid-containing peptide, causes Ca(2+)-dependent catecholamine secretion from adrenal medullary chromaffin cells.
We examined the effect of trichosporin-B-III, an alpha-aminoisobutyric acid-containing antibiotic peptide consisting of 19 amino acid residues and a phenylalaninol, on catecholamine secretion from cultured bovine adrenal chromaffin cells. Incubation of the cells with trichosporin-B-III (3-20 microM) caused an increase in the secretion of catecholamines. The secretion induced by trichosporin-B-III at low concentrations (3 and 5 microM) was completely dependent on external Ca2+, whereas that induced by higher concentrations (10 and 20 microM) was partly independent of Ca2+. Trichosporin-B-III at low concentration (5 microM) did not increase the release of lactate dehydrogenase, a marker enzyme of cytoplasm, from the cells. In contrast, the peptide at higher concentration (10 microM) increased the release of the enzyme. Trichosporin-B-III also caused both 45Ca2+ influx into the cells and an increase in the intracellular free Ca2+ concentration. The increases in catecholamine secretion and 45Ca2+ influx behaved similarly in relation to trichosporin-B-III concentration (3-10 microM). The time courses of the increases in secretion, 45Ca2+ influx, and intracellular free Ca2+ concentration induced by trichosporin-B-III were also quite similar. Trichosporin-B-III-induced (at 5 microM) secretion was not affected by the elimination of Na+ from the incubation medium or by the addition of tetrodotoxin, a blocker of highly selective voltage-dependent Na+ channels, or hexamethonium, a blocker of nicotinic acetylcholine receptors. On the other hand, both diltiazem (2-200 microM) and nicardipine (1-200 microM), blockers of voltage-dependent Ca2+ channels, inhibited the secretion induced by trichosporin-B-III (5 microM) in a concentration-dependent manner. Trichosporin-B-III-induced (at 5 microM) secretion also was suppressed by the addition of Mn2+ (5 mM) to the medium. The diltiazem (20 microM) inhibition of trichosporin-B-III-induced (at 5 microM) secretion was reversed by increasing the external Ca2+ concentration. These results indicate that trichosporin-B-III causes the secretion of catecholamines from bovine adrenal chromaffin cells by two mechanisms, Ca2+ dependent and Ca2+ independent (only at high concentrations of trichosporin-B-III). Furthermore, these results strongly suggest that trichosporin-B-III, in Ca(2+)-dependent secretion, activates endogenous voltage-dependent Ca2+ channels, or itself forms the channels in the membranes, and induces Ca2+ influx into the cells.


Influenza virus polymerase subunits co-evolve to ensure proper levels of dimerization of the heterotrimer.
The influenza A virus RNA-dependent RNA polymerase complex consists in three subunits, PB2, PB1 and PA, that perform transcription and replication of the viral genome through very distinct mechanisms. Biochemical and structural studies have revealed that the polymerase can adopt multiple conformations and form oligomers. However so far it remained unclear whether the available oligomeric crystal structures represent a functional state of the polymerase. Here we gained new insights into this question, by investigating the incompatibility between non-cognate subunits of influenza polymerase brought together through genetic reassortment. We observed that a 7:1 reassortant virus whose PB2 segment derives from the A/WSN/33 (WSN) virus in an otherwise A/PR/8/34 (PR8) backbone is attenuated, despite a 97% identity between the PR8-PB2 and WSN-PB2 proteins. Independent serial passages led to the selection of phenotypic revertants bearing distinct second-site mutations on PA, PB1 and/or PB2. The constellation of mutations present on one revertant virus was studied extensively using reverse genetics and cell-based reconstitution of the viral polymerase. The PA-E349K mutation appeared to play a major role in correcting the initial defect in replication (cRNA -> vRNA) of the PR8xWSN-PB2 reassortant. Strikingly the PA-E349K mutation, and also the PB2-G74R and PB1-K577G mutations present on other revertants, are located at a dimerization interface of the polymerase. All three restore wild-type-like polymerase activity in a minigenome assay while decreasing the level of polymerase dimerization. Overall, our data show that the polymerase subunits co-evolve to ensure not only optimal inter-subunit interactions within the heterotrimer, but also proper levels of dimerization of the heterotrimer which appears to be essential for efficient viral RNA replication. Our findings point to influenza polymerase dimerization as a feature that is controlled by a complex interplay of genetic determinants, can restrict genetic reassortment, and could become a target for antiviral drug development.


Metal-free coupling of azoles with 2- and 3-haloindoles providing access to novel 2- or 3-(azol-1-yl)indole derivatives.
Thermal or microwave-mediated heating of 2- or 3-haloindoles with azoles (pK(a) < 8) provides a straightforward, metal-free, and environmentally friendly access to novel 2-(azol-1-yl)indoles. Furthermore, previously unknown 2,3-bis(azolyl-1-yl)indoles can be prepared from 2,3-dihaloindoles by sequential reaction with two distinct azoles. This operationally simple acid-catalyzed process delivers novel indole derivatives in fair to excellent yields and expands the chemical diversity of substitutions that can be introduced on this medicinally important scaffold.


Daxx-beta and Daxx-gamma, two novel splice variants of the transcriptional co-repressor Daxx.
Daxx is involved in transcriptional control and apoptosis. It comprises several domains, including a regulatory C terminus that is responsible for the interaction with numerous proteins such as p53, promyelocytic leukemia protein (PML), and Hsp27. Here, we describe the identification and characterization of two novel variants of Daxx termed Daxx-β and Daxx-γ, which are generated by alternative splicing. Alternative splicing results in a truncated regulatory C terminus in both proteins. As a consequence, Daxx-β and Daxx-γ show a markedly decreased affinity to PML, which in turn is associated with a different subnuclear localization of these proteins compared with Daxx. Although Daxx is localized mainly in PML-oncogenic domains (PODs) Daxx-β and Daxx-γ display a distinct distribution pattern. Furthermore, Daxx-β and Daxx-γ show a decreased affinity to p53 also due to the truncated C terminus. We provide evidence that the p53 recruitment into PODs is Daxx isoform-dependent. The decreased affinity of Daxx-β/-γ to p53 and PML results in a diffuse localization of p53 throughout the nucleus. In contrast to Daxx, Daxx-β and Daxx-γ are unable to repress p53-mediated transcription. Therefore, alternative splicing of Daxx might indicate an additional level in the cellular apoptosis network.


[Asphyxia and hypoxic-ischaemic encephalopathy].
Hypoxic-ischaemic encephalopathy is a common cause of death and disability in newborns. Brain damage related to a perinatal insult is the result of a dynamic process, and its progressing development over time allows for specific interventions to reduce total damage, as described in this review. Despite therapeutic hypothermia which currently is the only treatment available, a considerable number of newborns still have adverse outcomes. Prognosis is evaluated by clinical examination and paraclinical investigations. There is still a need for novel treatments and better prognostic and diagnostic tools to improve outcome.


[Features of responses of the neurons of the caudate nucleus to various localizations of a photic stimulus in the visual field of the alert cat].
The responses of caudate neurons to restricted light stimuli differently located in the visual field were studied during natural fixation of the gaze on the visual target in awake cats. 71% of visually responsive neurons were sensitive to changing of stimulus location. It indicates that some neurons (17%) had restricted receptive fields in the centre of the gaze, other neurons (17%) had decreasing sensitivity to stimuli removed from the centre of the field and 37% of responsive neurons received information of different quality from different parts of the visual field. It is suggested that caudate nucleus neurons participate in the analysis of visual sensory information.


[Kinetics of thermal inactivation of peroxidases in turnips and peas].
The heat inactivation of peroxidase in green peas and turnips grown in Venezuela was determined experimentally at temperatures ranging from 60 degrees C to 76 degrees C for turnips, and between 72 degrees C and 82 degrees C for green peas. The thermal destruction curves obtained were found to be biphasic, following first-order kinetics of inactivation, and with average times for the change of phase of 22.5 sec and 23.8 sec for the turnips and peas, respectively. The activation energies, as calculated by the Arrhenius equation for the first and second inactivation intervals, were 27.1 kcal/mole and 43.5 kcal/mole for turnips, and 34.5 kcal/mole and 28.9 kcal/mole for green peas.


The effect of nitric oxide/endothelins system on the hepatic ischemia/reperfusion injury.
The relationship between the hepatic ischemia/reperfusion (I/R) injury and the balance of nitric oxide/endothelins (NO/ET) was studied. The changes of the ratio of NO/ET and the hepatic injury were observed in a rat hepatic I/R model pretreated with several tool drugs. In the acute phase of hepatic I/R injury, the ratio of plasma NO/ET was reduced from 1.58 +/- 0.20 to 0.29 +/- 0.05 (P < 0.01) and the hepatic damage deteriorated. NO donor L-Arg and ET receptor antagonist TAK-044 could alleviate the hepatic I/R injury to some degree, whereas NO synthase inhibitor L-NAME aggravated the damage. It was concluded that the hepatic I/R injury might be related with the disturbance of the NO/ET balance. Regulation of this balance might have an effect on the I/R injury.


Retinal polarization effects.
Retinal photographs taken with crossed polarizers in the input and recording light paths show a cross or brush pattern overlying the macula. Experiments were done indicating that this pattern is due to the birefringence in cone-photoreceptor outer segments. Haidinger's brushes are also attributed to cone outer segments, but the effect is due to a combination of the dispersion in the birefringence and the dichroism of the outer segments. The objective, polarized-light retinal pattern is shown to be a useful clinical tool for diagnosing diseases affecting the macula.


Oxidation of Reduced Ceria by Incorporation of Hydrogen.
The interaction of hydrogen with reduced ceria (CeO2-x ) powders and CeO2-x (111) thin films was studied using several characterization techniques including TEM, XRD, LEED, XPS, RPES, EELS, ESR, and TDS. The results clearly indicate that both in reduced ceria powders as well as in reduced single crystal ceria films hydrogen may form hydroxyls at the surface and hydride species below the surface. The formation of hydrides is clearly linked to the presence of oxygen vacancies and is accompanied by the transfer of an electron from a Ce3+ species to hydrogen, which results in the formation of Ce4+ , and thus in oxidation of ceria.


Serum from COVID-19 patients early in the pandemic shows limited evidence of cross-neutralization against variants of concern.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) results in a variety of clinical symptoms ranging from no or mild to severe disease. Currently, there are multiple postulated mechanisms that may push a moderate to severe disease into a critical state. Human serum contains abundant evidence of the immune status following infection. Cytokines, chemokines, and antibodies can be assayed to determine the extent to which a patient responded to a pathogen. We examined serum and plasma from a cohort of patients infected with SARS-CoV-2 early in the pandemic and compared them to negative-control sera. Cytokine and chemokine concentrations varied depending on the severity of infection, and antibody responses were significantly increased in severe cases compared to mild to moderate infections. Neutralization data revealed that patients with high titers against an early 2020 isolate had detectable but limited neutralizing antibodies against newly circulating SARS-CoV-2 variants of concern. This study highlights the potential of re-infection for recovered COVID-19 patients.


Administering an appeasing substance to beef calves at weaning to optimize productive and health responses during a 42-d preconditioning program.
This experiment evaluated the impacts of administering a bovine appeasing substance (BAS) to beef calves at weaning on their performance, physiological responses, and behavior during a 42-d preconditioning program. Eighty calves (40 heifers and 40 steers; 90% British × 10% Nellore) were weaned at 233 ± 2 d of age (day 0); ranked by sex, weaning age, and body weight (BW); and assigned to receive BAS (IRSEA Group, Quartier Salignan, France; n = 40) or placebo (diethylene glycol monoethyl ether; CON; n = 40). Treatments (5 mL) were topically applied to the nuchal skin area of each animal following dam separation. Within treatment, calves were allocated to one of eight drylot pens (four pens per treatment; pen being the experimental unit) and received a free-choice total mixed ration (TMR) from day 0 to 42, intake of which was assessed daily. Live behavior observations were conducted on days 1, 2, 4, 8, 16, and 32. Temperament was assessed and blood samples were collected via jugular venipuncture on days -21, 0, 3, 7, 14, 28, and 42. Hair samples were collected from the tail switch on days 0, 14, 28, and 42. Calves were vaccinated against bovine respiratory disease viruses on days -21 and 0. Average daily gain from day 0 to 42 did not differ between treatments (P = 0.57) but was greater (P = 0.05) in BAS vs. CON calves from day 0 to 28. Intake of TMR was greater (P = 0.05) during the first week for BAS vs. CON calves (treatment × week; P = 0.08). The mean proportion of calves feeding simultaneously and performance of social and play behaviors were greater (P ≤ 0.05) for BAS vs. CON calves. Escape attempts were greater (P < 0.01) for BAS vs. CON calves on day 1 (treatment × day; P = 0.03). Exit velocity was greater (P = 0.04) for CON vs. BAS calves on day 14 and tended (P = 0.10) to be greater for CON vs. BAS calves on day 7 (treatment × day; P = 0.03). Mean plasma concentrations of haptoglobin were greater (P = 0.02) in CON vs. BAS calves. Hair cortisol concentrations were greater (P = 0.05) in CON vs. BAS calves on day 14 (treatment × day; P = 0.03). Mean serum concentrations of antibodies against bovine viral diarrhea virus were greater (P = 0.02) in BAS vs. CON calves. Collectively, BAS administration to beef calves at weaning alleviated stress-induced physiological reactions, improved temperament evaluated via chute exit velocity, enhanced humoral immunity acquired from vaccination, and appeared to have accelerated adaptation to novel management scheme and environment.


Antidiabetic treatment restores adiponectin serum levels and APPL1 expression, but does not improve adiponectin-induced vasodilation and endothelial dysfunction in Zucker diabetic fatty rats.
Adiponectin is able to induce NO-dependent vasodilation in Zucker lean (ZL) rats, but this effect is clearly alleviated in their diabetic littermates, the Zucker diabetic fatty (ZDF) rats. ZDF rats also exhibit hypoadiponectinemia and a suppressed expression of APPL1, an adaptor protein of the adiponectin receptors, in mesenteric resistance arteries. Whether an antidiabetic treatment can restore the vasodilatory effect of adiponectin and improve endothelial function in diabetes mellitus type 2 is not known. During our animal experiment from week 11 to 22 in each case seven ZDF rats received an antidiabetic treatment with either insulin (ZDF+I) or metformin (ZDF+M). Six normoglycemic ZL and six untreated ZDF rats served as controls. Blood glucose was measured at least weekly and serum adiponectin levels were quantified via ELISA in week 11 and 22. The direct vasodilatory response of their isolated mesenteric resistance arteries to adiponectin as well as the endothelium-dependent and -independent function was evaluated in a small vessel myograph. Additionally, the expression of different components of the adiponectin signaling pathway in the resistance arteries was quantified by real-time RT-PCR. In ZDF rats a sufficient blood glucose control could only be reached by treatment with insulin, but both treatments restored the serum levels of adiponectin and the expression of APPL1 in small resistance arteries. Nevertheless, both therapies were not able to improve the vasodilatory response to adiponectin as well as endothelial function in ZDF rats. Concurrently, a downregulation of the adiponectin receptors 1 and 2 as well as endothelial NO-synthase expression was detected in insulin-treated ZDF rats. Metformin-treated ZDF rats showed a reduced expression of adiponectin receptor 2. An antidiabetic treatment with either insulin or metformin in ZDF rats inhibits the development of hypoadiponectinemia and downregulation of APPL1 in mesenteric resistance arteries, but is not able to improve adiponectin induced vasodilation and endothelial dysfunction. This is possibly due to alterations in the expression of adiponectin receptors and eNOS.


Modification of acute murine cytomegalovirus adrenal gland infection by adoptive spleen cell transfer.
Homozygous (nu/nu), athymic nude mice, infected with murine cytomegalovirus (MCMV), develop unremitting and ablative virus infection involving both the adrenal cortex and medulla. During acute infection, adoptive transfer of MCMV-immune, but not naive, spleen cells suppressed virus replication in the adrenal glands, but not the lungs or salivary gland. T lymphocytes, not macrophages or B cells, were responsible for limiting viral replication. The effect by donor cells was restricted by compatibility at the major histocompatibility locus. Restriction of MCMV replication in the adrenal gland was associated with T lymphocytes of the L3T4 phenotype. Thus, T-cell immunity is critical in regulating MCMV replication in the adrenal glands, and T lymphocytes restricted by class II major histocompatibility antigens mediate this effect.


In vivo administration of purified human interleukin-2 to patients with cancer: development of interleukin-2 receptor positive cells and circulating soluble interleukin-2 receptors following interleukin-2 administration.
Recent studies have demonstrated efficacy of immunotherapies including interleukin-2 (IL-2) in the treatment of malignancies in rodents and humans. High levels of IL-2 receptor-positive cells were found in the peripheral blood of patients receiving recombinant IL-2 in these Phase I clinical trials. This was demonstrated both in patients receiving i.v. IL-2 who had detectable circulating levels of IL-2 as well as in patients receiving i.p. IL-2 who did not. Up to 100% of the anti-Tac binding could be inhibited by preincubation with IL-2 indicating that this was indeed an IL-2 receptor that was identified. Two-color experiments demonstrated that few Leu 2-positive cells (less than 5-10%) but over 30% of the Leu 3-positive cells bore Tac antigen. Most of the M3-positive monocytes were Tac positive (83.7%) and negative for other T-cell (Leu-4) and nonspecific murine markers (Lyt-2 and Thy 1.2). Although normal individuals had a mean of only 186 units/ml (range, 83-335 units/ml) of soluble IL-2 receptor, patients receiving IL-2 had as much as 20,000 units/ml of soluble IL-2 receptor line in their serum. The physiological role of the IL-2 receptor identified on the cell surface of Leu 3 and M3-positive cells as well as in the serum is unclear. Soluble IL-2 receptors appeared in the circulation early following IL-2 administration, approximately 1 week prior to the detection of circulating IL-2 receptor-bearing cells. Further studies will be needed to assess the role of IL-2 in monocyte function, the precise function of IL-2 receptor-bearing Leu 3-positive cells, and the relationship of these findings to the toxicity and success of this immunotherapy in humans.


Molecular mechanics calculations of proteins. Comparison of different energy minimization strategies.
A general strategy for performing energy minimization of proteins using the SYBYL molecular modelling program has been developed. The influence of several variables including energy minimization procedure, solvation, dielectric function and dielectric constant have been investigated in order to develop a general method, which is capable of producing high quality protein structures. Avian pancreatic polypeptide (APP) and bovine pancreatic phospholipase A2 (BP PLA2) were selected for the calculations, because high quality X-ray structures exist and because all classes of secondary structure are represented in the structures. The energy minimized structures were evaluated relative to the corresponding X-ray structures. The overall similarity was checked by calculating RMS distances for all atom positions. Backbone conformation was checked by Ramachandran plots and secondary structure elements evaluated by the length on hydrogen bonds. The dimensions of active site in BP PLA2 is very dependent on electrostatic interactions, due to the presence of the positively charged calcium ion. Thus, the distances between calcium and the calcium-coordinating groups were used as a quality index for this protein. Energy minimized structures of the trimeric PLA2 from Indian cobra (N.n.n. PLA2) were used for assessing the impact of protein-protein interactions. Based on the above mentioned criteria, it could be concluded that using the following conditions: Dielectric constant epsilon = 4 or 20; a distance dependent dielectric function and stepwise energy minimization, it is possible to reproduce X-ray structures very accurately without including explicit solvent molecules.


Molecular analysis of endocrine disruption in hornyhead turbot at wastewater outfalls in southern california using a second generation multi-species microarray.
Sentinel fish hornyhead turbot (Pleuronichthysverticalis) captured near wastewater outfalls are used for monitoring exposure to industrial and agricultural chemicals of ~ 20 million people living in coastal Southern California. Although analyses of hormones in blood and organ morphology and histology are useful for assessing contaminant exposure, there is a need for quantitative and sensitive molecular measurements, since contaminants of emerging concern are known to produce subtle effects. We developed a second generation multi-species microarray with expanded content and sensitivity to investigate endocrine disruption in turbot captured near wastewater outfalls in San Diego, Orange County and Los Angeles California. Analysis of expression of genes involved in hormone [e.g., estrogen, androgen, thyroid] responses and xenobiotic metabolism in turbot livers was correlated with a series of phenotypic end points. Molecular analyses of turbot livers uncovered altered expression of vitellogenin and zona pellucida protein, indicating exposure to one or more estrogenic chemicals, as well as, alterations in cytochrome P450 (CYP) 1A, CYP3A and glutathione S-transferase-α indicating induction of the detoxification response. Molecular responses indicative of exposure to endocrine disruptors were observed in field-caught hornyhead turbot captured in Southern California demonstrating the utility of molecular methods for monitoring environmental chemicals in wastewater outfalls. Moreover, this approach can be adapted to monitor other sites for contaminants of emerging concern in other fish species for which there are few available gene sequences.


Metabolic Regulation of Copper Toxicity during Marine Mussel Embryogenesis.
The development of new tools for assessing the health of cultured shellfish larvae is crucial for aquaculture industries to develop and refine hatchery methodologies. We established a large-volume ecotoxicology/health stressor trial, exposing mussel (Perna canaliculus) embryos to copper in the presence of ethylenediaminetetraacetic acid (EDTA). GC/MS-based metabolomics was applied to identify potential biomarkers for monitoring embryonic/larval health and to characterise mechanisms of metal toxicity. Cellular viability, developmental abnormalities, larval behaviour, mortality, and a targeted analysis of proteins involved in the regulation of reactive oxygen species were simultaneously evaluated to provide a complementary framework for interpretative purposes and authenticate the metabolomics data. Trace metal analysis and speciation modelling verified EDTA as an effective copper chelator. Toxicity thresholds for P. canaliculus were low, with 10% developmental abnormalities in D-stage larvae being recorded upon exposure to 1.10 μg·L-1 bioavailable copper for 66 h. Sublethal levels of bioavailable copper (0.04 and 1.10 μg·L-1) caused coordinated fluctuations in metabolite profiles, which were dependent on development stage, treatment level, and exposure duration. Larvae appeared to successfully employ various mechanisms involving the biosynthesis of antioxidants and a restructuring of energy-related metabolism to alleviate the toxic effects of copper on cells and developing tissues. These results suggest that regulation of trace metal-induced toxicity is tightly linked with metabolism during the early ontogenic development of marine mussels. Lethal-level bioavailable copper (50.3 μg·L-1) caused severe metabolic dysregulation after 3 h of exposure, which worsened with time, substantially delayed embryonic development, induced critical oxidative damage, initiated the apoptotic pathway, and resulted in cell/organism death shortly after 18 h of exposure. Metabolite profiling is a useful approach to (1) assess the health status of marine invertebrate embryos and larvae, (2) detect early warning biomarkers for trace metal contamination, and (3) identify novel regulatory mechanisms of copper-induced toxicity.


Strain Stiffening Hydrogels through Self-Assembly and Covalent Fixation of Semi-Flexible Fibers.
Biomimetic, strain-stiffening materials are reported, made through self-assembly and covalent fixation of small building blocks to form fibrous hydrogels that are able to stiffen by an order of magnitude in response to applied stress. The gels consist of semi-flexible rodlike micelles of bisurea bolaamphiphiles with oligo(ethylene oxide) (EO) outer blocks and a polydiacetylene (PDA) backbone. The micelles are fibers, composed of 9-10 ribbons. A gelation method based on Cu-catalyzed azide-alkyne cycloaddition (CuAAC), was developed and shown to lead to strain-stiffening hydrogels with unusual, yet universal, linear and nonlinear stress-strain response. Upon gelation, the X-ray scattering profile is unchanged, suggesting that crosslinks are formed at random positions along the fiber contour without fiber bundling. The work expands current knowledge about the design principles and chemistries needed to achieve fully synthetic, biomimetic soft matter with on-demand, targeted mechanical properties.


[Cardiovascular risk in autoimmune and inflammatory disorders].
Atherosclerosis (ATS) is characterized by an inflammatory process initiated by oxidized LDL in the vessel wall, where activation of cells of the innate and adaptive immune system takes place. ATS is accelerated with an increased risk of cardiovascular (CV) events in systemic autoimmune disorders (AID) such as systemic lupus erythematosus or rheumatoid arthritis (RA). In addition to the traditional CV risk factors, which are over-represented in AID, the underlying chronic inflammation and dysregulation of the immune system play an amplifying role in ATS. Although certain drugs used in AID can increase the CV risk, the control of the disease as permitted by TNF-blocking agents in RA, reduces this risk. The strategies specific to AID to reduce the CV risk remain to be better defined.


PHASE: a randomised, controlled trial of supervised self-help cognitive behavioural therapy in primary care.
Common mental health problems account for up to 40% of all general practitioner (GP) consultations. Patients have limited access to evidence-based psychological therapies. Cognitive behavioural therapy self-help strategies offer one potential solution. To determine differences in clinical outcome, patient satisfaction and costs, between a cognitive behavioural-based self-help package facilitated by practice nurses compared to ordinary care by GPs for mild to moderate anxiety and depression. Randomised controlled trial. Seventeen primary healthcare teams. Patients presenting to their GP with mild to moderate anxiety and/or depression were recruited to the study and randomised to receive either a self-help intervention facilitated by practice nurses or ordinary care. The self-help intervention consisted of up to three appointments: two 1 week apart and a third 3 months later. There were no restrictions on ordinary care. Intention-to-treat analysis showed that patients treated with practice nurse-supported cognitive behavioural therapy self-help attained similar clinical outcomes for similar costs and were more satisfied than patients treated by GPs with ordinary care. On-treatment analysis showed patients receiving the facilitated cognitive behavioural therapy self-help were more likely to be below clinical threshold at 1 month compared to the ordinary care group (odds ratio [OR] = 3.65, 95% confidence interval [CI] = 1.87 to 4.37). This difference was less well marked at 3 months (OR = 1.36, 95% CI = 0.52 to 3.56). Facilitated cognitive behavioural self-help may provide a short-term cost-effective clinical benefit for patients with mild to moderate anxiety and depression. This has the potential to help primary care provide a choice of effective psychological as well as pharmacological treatments for mental health problems.


Biomechanical responses to repeated stretches in human hamstring muscle in vivo.
To examine stiffness, energy, and passive torque in the dynamic and static phases of a stretch maneuver in the human hamstring muscle in vivo we used a test-retest protocol and a repeated stretches protocol. Resistance to stretch was defined as passive torque (in newton-meters) offered by the hamstring muscle group during passive knee extension as measured using an isokinetic dynamometer with a modified thigh pad. In 13 uninjured subjects, the knee was passively extended to a predetermined final position (0.0875 rad/ sec, dynamic phase) where it remained stationary for 90 seconds (static phase). The test-retest protocol included two tests administered 1 hour apart. On a separate occasion, five consecutive static stretches were administered separated by 30 seconds and followed by a sixth stretch 1 hour later. For the test-retest phase, stiffness and energy in the dynamic phase and passive torque in the static phase did not differ and yielded correlations of r = 0.91 to 0.99. During the static phase, passive torque declined in both tests (P < 0.0001). For the repeated stretches, decreases were observed for energy (P < 0.01) and stiffness (P < 0.05) in the dynamic phase and for passive torque (P < 0.0001) in the static phase. However, the decline in the variables returned to baseline within 1 hour. The data show that the method employed is a useful tool for measuring biomechanical variables during a stretch maneuver. This may provide a more detailed method to examine skeletal muscle flexibility.


Stress and Western diets increase vulnerability to neuropsychiatric disorders: A common mechanism.
In modern lifestyle, stress and Western diets are two major environmental risk factors involved in the etiology of neuropsychiatric disorders. Lifelong interactions between stress, Western diets, and how they can affect brain physiology, remain unknown. A possible relation between dietary long chain polyunsaturated fatty acids (PUFA), endocannabinoids, and stress is proposed. This review suggests that both Western diets and negative stress or distress increase n-6/n-3 PUFA ratio in the phospholipids of the plasma membrane in neurons, allowing an over-activation of the endocannabinoid system in the limbic areas that control emotions. As a consequence, an excitatory/inhibitory imbalance is induced, which may affect the ability to synchronize brain areas involved in the control of stress responses. These alterations increase vulnerability to neuropsychiatric disorders. Accordingly, dietary intake of n-3 PUFA would counter the effects of stress on the brain of stressed subjects. In conclusion, this article proposes that PUFA, endocannabinoids, and stress form a unique system which is self-regulated in limbic areas which in turn controls the effects of stress on the brain throughout a lifetime.


Influence of extracellular [Ca2+] on secretory and redox responses to CCK-8 in perfused rat pancreas.
There is a distinct discrepancy between the dose-dependent secretory responses (pancreatic protein output and juice flow) and the dose-dependent redox responses of cytochromes aa3, b, and c+c1 to various concentrations of CCK-8: a bell-shaped relation for the secretory responses contrasts with a sigmoidal relation for the redox responses. Continuous stimulation with CCK-8 at physiological low concentrations (5-10 pM) induced an increase in pancreatic protein output with little if any reduction of cytochromes. Continuous stimulation with CCK-8 at a pharmacological intermediate concentration (50 pM), however, induced an increase in pancreatic protein output with delayed reduction of cytochromes. The secretory and redox responses were completely abolished when CaCl2 was removed from the perfusing and bathing solution. An almost linear relation was found between the magnitude of the protein output in response to 50 pM CCK-8 and extracellular [Ca2+] [( Ca2+]o) in the range of 0.5-2.5 mM. Similar relations were also found between the levels of reduction of cytochromes and [Ca2+]o. These results are compatible with the view that continuous stimulation with CCK-8 at a pharmacological or a pathological concentration may cause excess increase in intracellular [Ca2+] and intramitochondrial Ca2+ and in this way may accelerate the formation of reducing power for oxidative phosphorylation.


Assessment and treatment of the child with mental retardation. Guidelines for the public school therapist.
Physical therapists employed in public schools may be responsible for the evaluation and treatment of not only physically handicapped children, but also children who have moderate to severe motor disabilities secondary to mental retardation. The purpose of this article is to suggest appropriate assessment and treatment techniques for these children. General principles of intervention based on neurophysiologic treatment approaches, particularly sensory integration, are described. Examples of specific assessment and treatment strategies are given for visual, auditory, tactile, olfactory-gustatory, proprioceptive-kinesthetic, and vestibular functions. In addition, self-stimulatory behaviors, tests of motor and reflex development, problems in muscle tone and strength, and variations in gait patterns are discussed.


Novel complex polar lipids from the methanogenic archaebacterium Methanospirillum hungatei.
The methanogenic archaebacterium Methanospirillum hungatei contains two unusual phosphoglycolipids that account for 64 percent of the total cellular lipids. These lipids are derivatives of the dibiphytanyl diglycerol tetraether, previously identified in methanogens. One of the free hydroxyls of this tetraether is esterified with glycerophosphoric acid, and the other is linked glycosidically to a disaccharide. The two phosphoglycolipids may function as covalently bonded lipid bilayers to impart stability and rigidity to methanogen membranes.


Cd52, known as a major maturation-associated sperm membrane antigen secreted from the epididymis, is not required for fertilization in the mouse.
CD52 is a glycosylphosphatidylinositol (GPI)-anchored antigen expressed on lymphocytes and in epididymal epithelial cells. CD52 is also known as "maturation-associated sperm antigen" but its function is unknown. We therefore generated Cd52 disrupted mice. The resulting Cd52 null mice were healthy, even though Cd52 is expressed on cells of the immune system. We then examined a possible role for CD52 in reproduction. Sperm from Cd52-deficient males were investigated and the viability, motility, morphology, and incidence of spontaneous acrosome reactions were found to be all similar to values for wild-type sperm. In in vitro fertilization system, the sperm showed normal fertilizing ability. As CD52 was found to be transferred onto sperm only after they had migrated into the vas deferens, we examined the behavior of sperm from Cd52-deficient mice in vivo. The mice mated naturally and we observed that a normal number of sperm passed through the uterotubal junction, known to the crucial hurdle for various gene knockouts resulting in infertile sperm. As a consequence, there was no difference in the litter size from the wild-type and Cd52-null males. Our results therefore indicate CD52 is not required for fertilization in the mouse either in vivo or in vitro.


A novel antitumor activity of deguelin targeting the insulin-like growth factor (IGF) receptor pathway via up-regulation of IGF-binding protein-3 expression in breast cancer.
In this study, we investigated the antitumor effects of deguelin in several human breast cancer cells in vitro and in vivo. Deguelin inhibited cell viability and the anchorage-dependent and anchorage-independent colony formation of triple-negative (MDA-MB-231 and MDA-MB-468) and triple-positive (MCF-7) breast cancer cells, and it significantly reduced the growth of MCF-7 cell xenograft tumors. The induction of apoptosis, inhibition of insulin-like growth factor-1 receptor (IGF-1R) signaling activation, and up-regulation of IGF-binding protein-3 (IGFBP-3) expression may be associated with deguelin-mediated antitumor effects. Our findings suggest a potential therapeutic use for deguelin in patients with triple-negative breast cancer and for those with breast cancers who are sensitive to endocrine- and HER2-targeted therapies.


A rapid water bath PCR combined with lateral flow assay for the simultaneous detection of SARS-CoV-2 and influenza B virus.
The outbreak of the coronavirus disease 2019 caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has resulted in significant global health and economic threats to the human society. Thus, a rapid and accurate detection method for early testing and diagnosis should be established. In this study, a rapid water bath polymerase chain reaction (PCR) combined with lateral flow assay was developed to detect SARS-CoV-2 and influenza B virus simultaneously. A homemade automated transfer device equipped with reaction tube shuttled rapidly between two water baths at 98 °C and 53 °C to realize rapid PCR. After amplification, two-ended labeled PCR products were detected using the lateral flow strip with two test lines and streptavidin-conjugated quantum dot nanobeads. The fluorescence value was read using a handheld instrument. The established assay could complete reverse-transcription PCR amplification and lateral flow detection in 45 minutes. The detection limits were 8.44 copies per μL and 14.23 copies per μL for SARS-CoV-2 and influenza B virus, respectively. The coefficients of variation of the test strip were 10.10% for the SARS-CoV-2 and 4.94% for the influenza B virus, demonstrating the excellent repeatability of the experiment. These results indicated that the rapid PCR combined with lateral flow assay could detect SARS-CoV-2 and influenza B virus simultaneously at a short assay time and low cost, thereby showing the remarkable potential for the rapid and multiplex detection of respiratory viruses in resource-limited settings.


Lead poisoning mimicking acute porphyria!
We are presenting a case of a 13-year-old autistic boy whose urine porphyrin test came positive on three separate occasions. The child was brought to emergency department of Kasturba Medical College Hospital, Attavar, Mangalore, India, with fever and acute abdominal pain, with no previous history of any serious illness. Investigations revealed thalassemia trait,microcytic hypochromic anaemia while the other biochemical and haematological parameters were normal. False positive urine porphyrin test may be seen in porphyria induced by liver cancer, hepatitis and heavy metal poisoning such as lead, arsenic and mercury. Blood lead (PbB) level was 59.5μg/dl. Further evaluation revealed a daily consumption of native medicine in the form of syrup.


Visual crowding is anisotropic along the horizontal meridian during smooth pursuit.
Humans make smooth pursuit eye movements to foveate moving objects of interest. It is known that smooth pursuit alters visual processing, but there is currently no consensus on whether changes in vision are contingent on the direction the eyes are moving. We recently showed that visual crowding can be used as a sensitive measure of changes in visual processing, resulting from involvement of the saccadic eye movement system. The present paper extends these results by examining the effect of smooth pursuit eye movements on the spatial extent of visual crowding-the area over which visual stimuli are integrated. We found systematic changes in crowding that depended on the direction of pursuit and the distance of stimuli from the pursuit target. Relative to when no eye movement was made, the spatial extent of crowding increased for objects located contraversive to the direction of pursuit at an eccentricity of approximately 3°. By contrast, crowding for objects located ipsiversive to the direction of pursuit remained unchanged. There was no change in crowding during smooth pursuit for objects located approximately 7° from the fovea. The increased size of the crowding zone for the contraversive direction may be related to the distance that the fovea lags behind the pursuit target during smooth eye movements. Overall, our results reveal that visual perception is altered dynamically according to the intended destination of oculomotor commands.


[Her-2/neu, Ki-67 expression and ploidy in breast carcinoma].
Expression of the markers was studied immunocytochemically on cytological preparations of breast tumor, ploidy was assessed by flow cytofluorimetry. The material was divided into 2 groups: diploid--41.4% cases and aneuploid--58.6%. Hyperexpression of Her-2/neu in the first group was observed in 54.5%, in the 2nd group in 48.6% cases. Expression of Ki-67 in 63.4% and in 68.9%, respectively. Tumours with high proliferative activity were numerous in aneuploid tissues, and in diploid tumours moderate activity prevailed (p = 0.006). Direct correlation between the markers was observed, high expression of Ki-67 more frequently was associated with positive expression of Her-2/neu. Thus, aneuploid tumours with high proliferative activity and hyperexpression of Her-2/neu are more aggressive tumours of a large size.


Homologous regulation of gonadotropin-releasing hormone receptors in cultured pituitary cells.
Specific receptors for gonadotropin-releasing hormone (GnRH) in cultured rat pituitary cells were increased by subnanomolar concentrations of GnRH agonists and decreased by high concentrations of these peptides. The antagonist [D-Phe2, Pro3, D-Phe6]GnRH did not alter GnRH binding capacity and blocked the increase in sites induced by GnRH. These findings provide direct evidence for the homologous regulation of GnRH receptors by physiological concentrations of the hypothalamic peptide, an action that could mediate the cyclical and postcastration increases in GnRH receptors and responsiveness of the pituitary gonadotrophs.


The role of nickel accumulation and epithelial cell proliferation in orthodontic treatment-induced gingival overgrowth.
The aim of this study was to investigate the role of nickel in orthodontic treatment-induced gingival hyperplasia. The nickel concentration in gingival tissues with and without overgrowth, histopathology of gingival overgrowth, and epithelial cell proliferation response to different nickel concentrations were analysed. Ten patients receiving orthodontic therapy (eight females and two males, mean age 15.4 years) were included in the study. Hyperplastic and healthy gingiva samples were collected from the same patients. The amount of nickel in the gingival tissue samples was analysed using the atomic absorption spectrometry technique. The tissues removed from hyperplastic areas during gingivectomy were also used for histological analysis. To analyse the effect of nickel on epithelial cell proliferation, four different nickel concentrations (0.5, 2, 5, and 10 microg) were incubated with keratinocyte cells for 11 days. Mann-Whitney U-test, analysis of variance, and Tukey's test were used in the statistical analyses. The results did not show any difference in nickel concentration between the study and control gingiva tissue samples, but histological analysis demonstrated an increase in epithelial thickness and a significant increase (P = 0.031, 0.02, 0.02) in epithelial cell proliferation in response to low-dose nickel concentrations, with a toxic response to a higher dose. In the limitations of this study, it is plausible that the effect of a continuing low-dose nickel release to epithelium is the initiating factor of gingival overgrowth induced by orthodontic treatment.