Unlocking nature's (sub)cellular symphony: Phase separation in plant meristems.
Plant development is based on the balance of stem cell maintenance and differentiation in the shoot and root meristems. The necessary cell fate decisions are regulated by intricate networks of proteins and biomolecules within plant cells and require robust and dynamic compartmentalization strategies, including liquid-liquid phase separation (LLPS), which allows the formation of membrane-less compartments. This review summarizes the current knowledge about the emerging field of LLPS in plant development, with a particular focus on the shoot and root meristems. LLPS regulates not only floral transition and flowering time while integrating environmental signals in the shoots but also influences auxin signalling and is putatively involved in maintaining the stem cell niche (SCN) in the roots. Therefore, LLPS has the potential to play a crucial role in the plasticity of plant development, necessitating further research for a comprehensive understanding.


Blastogenic response of activated human umbilical cord blood T-lymphocytes.
Donor T-lymphocytes are thought to play a crucial role in both acute and chronic graft-versus-host disease (GvHD), pathological conditions that frequently complicate allogeneic bone marrow transplantation. These diseases are described as occurring with a lower incidence and lesser severity when human umbilical cord blood (HUCB) cells, which have recently emerged as a potential source of hematopoietic progenitors, are used for transplantation. This condition is probably related to the immaturity of neonatal HUCB T cells. Lymphocyte blastogenic response to phytohemagglutinin (PHA), evaluated by means of flow cytometry, is a useful tool for testing the functional ability of T-cells to display an immune response against allo-antigens, reproducing in vitro the in vivo mechanism of activation. This study was designed to verify whether an impairment in HUCB T-cell ability to undergo an in vitro blastogenic response to mitogens could explain their reduced in vivo ability to induce GvHD.


DNA methylation changes in cystic fibrosis: Cause or consequence?
Twin and sibling studies have shown that lung disease severity is variable among cystic fibrosis (CF) patients and affected to the same extent by genetic and nonheritable factors. Genetic factors have been thoroughly assessed, whereas the molecular mechanisms whereby nonheritable factors contribute to the phenotypic variability of CF patients are still unknown. Epigenetic modifications may represent the missing link between nonheritable factors and phenotypic variation in CF. Herein, we review recent studies showing that DNA methylation is altered in CF and we address three possible factors responsible for these variations: (i) overproduction of reactive oxygen species, (ii) depletion of DNA methylation cofactors and (iii) susceptibility to acute and chronic bacterial infections. Also, we hypothesize that the unique DNA methylation profile of each patient can modulate the phenotype and discuss the interest of implementing integrated genomic, epigenomic and transcriptomic studies to further understand the clinical diversity of CF patients (Graphical Abstract).


Layer I neurons of the rat neocortex. II. Voltage-dependent outward currents.
1. Whole cell patch-clamp techniques, combined with direct visualization of neurons, were used to study voltage-dependent potassium currents in layer 1 neurons and layer II/III pyramidal cells. 2. In the presence of tetrodotoxin, step depolarizations evoked an outward current. This current had a complex waveform and appeared to be a composite of early and late components. The early peak of the composite K+ outward current was larger in layer I neurons. 3. In both layer I and pyramidal cells, the composite outward K+ current could be separated into two components based on kinetic and pharmacological properties. The early component was termed I(A) because it was a transient outward current activating rapidly and then decaying. I(A) was more sensitive to blocking by 4-aminopyridine (4-AP) than tetraethylammonium (TEA). The second component, termed the delayed rectifier or I(DR), activated relatively slowly and did not decay significantly during a 200-ms test pulse. I(DR) was insensitive to blocking by 4-AP at concentrations up to 4 mM and blocked by > 60% by 40-60 mM TEA. 4. I(A) kinetics were examined in the presence of 40-60 mM TEA. Under these conditions, I(A) began to activate between -40 and -30 mV. Half-maximal activation occurred around 0 mV. In both layer I and pyramidal cells, the half-inactivation potential (Vh-inact) was around or more positive than -50 mV. At -60 mV, > 70% of I(A) conductance was available. I(A) decayed along a single exponential time course with a time constant of approximately 15 ms. This decay showed little voltage dependence. 5. In both layer I and pyramidal cells, I(DR) was studied in the presence of 4 mM 4-AP to block I(A) and in saline containing 0.2 mM Ca2+ and 3.6 mM Mg2+ to reduce contributions from Ca2+-dependent K+ currents. Under these conditions, I(DR) began to activate at -35 to -25 mV with Vh-act of 3.6 +/- 4.5 mV (mean +/- SD). The 10-90% rise time of I(DR) was 15 ms at 30 mV. At 2.2 ms after the onset of the command potential to +30 mV, I(DR) could reach a significant amplitude (approximately 1.5 nA in layer I neurons and 2.2 nA in pyramidal cells depending on the cell size). When long test pulses (> or = 1,000 ms) were used, a decay time constant approximately 800 ms at +40 mV was observed. In both layer I and pyramidal cells, steady state inactivation of I(DR) was minimal. 6. These results indicate that I(A) and I(DR) are the two major hyperpolarizing currents in layer I and pyramidal cells. The kinetics and pharmacological properties of I(A) and I(DR) were not significantly different in fast-spiking layer I neurons and regular-spiking layer II/III pyramidal cells. The relatively positive activation threshold (more than or equal to -40 mV) of both I(A) and I(DR) suggest that they do not play a role in neuronal behavior below action potential (AP) threshold and that their properties are more suitable to repolarize AP. The greater density of I(A) in layer I neurons appears responsible for fast spike generation.


Diabetic retinopathy and nephropathy in Fiji: comparison with data from an Australian diabetes centre.
As part of a project to improve diabetes care in Fiji, we assessed the magnitude of problems posed by diabetic retinopathy in that country and compared the findings with those from an Australian diabetes centre. The relationship between diabetic retinopathy and nephropathy was also examined in a subset of patients. A medical team from Australia screened a total of 446 type 2 diabetic patients (ethnicity: Fijian/Indian 16/84%) for diabetic retinopathy in five towns from the Division of Viti Levu, Fiji. The findings were compared with data obtained from 1659 type 2 diabetic patients who had attended an Australian diabetes centre (ethnicity Indian/ Anglo-Celtic 12/88%). In both cohorts, retinopathy was assessed by direct fundoscopy and a spot urine sample was collected for determination of albuminuria (defined as a concentration > 50 mg/L). The prevalence of diabetic retinopathy increased linearly with duration of diabetes. It was higher in Fiji, even when cases from the same ethnicity (i.e. Indians) and duration were compared (P < 0.05). Extrapolation of the data points suggests a delay in the diagnosis of diabetes in Fiji. Of those patients with retinopathy in Fiji, more than half had moderate to severe non-proliferative diabetic retinopathy or proliferative diabetic retinopathy, significantly higher than patients in the Australian cohort (chi2 = 29.2; P < 0.0001). Retinopathy was not a predictor of albuminuria in Fijian Indians (chi2 = 0.4; P = 0.5). In contrast, Australian Indians with retinopathy had significantly more albuminuria (chi2 = 10.2; P = 0.001). Severe diabetic retinopathy is common in both ethnic groups in Fiji. A delay in the diagnosis of diabetes as well as poor glycaemic control are possible factors. The availability of laser therapy is important to prevent loss of vision, but it is also essential that appropriate training of health professionals is integrated with a programme of diabetic complication screening to support this form of therapy.


Ascaridia galli - An old problem that requires new solutions.
Reports of Ascaridia galli in laying hens in Europe have increased since the ban on conventional battery cages in 2012. As this parasite is transmitted directly via the faecal-oral route by parasite eggs containing a larva, it is reasonable to assume that the escalating problem is related to the increased exposure now occurring in modern welfare-friendly cage-free housing systems. On many farms, A. galli reappears in subsequent flocks, even though the birds have no access to the outdoors, biosecurity is high and empty houses are cleaned and disinfected during downtime. Since the egg production cycle lasts only ≈80 weeks and recombinant antigen production for helminth vaccines has not yet been solved, the development of a vaccine seems to be an unrealistic option. Therefore, disrupting the life cycle of the parasite by other means, including the strategic use of dewormers, appears to be the key to controlling infection. Of concern is that only one class of anthelmintics is licenced for poultry in Europe and that are usually administered indiscriminately through the birds' drinking water and often too late when the parasite is already established. If current calendar-based parasite control strategies are not changed, there is a risk that resistance to anthelmintics may develop, as has already been demonstrated with nematodes in livestock. We insist that treatments can be more effective and the risk of developing drug resistance can be mitigated if we invest in a better understanding of A. galli responses to more prudent and judicious use of anthelmintics. This review identifies knowledge gaps and highlights aspects of sustainable parasite control that require further research to support commercial egg producers.


Postoperative dyspnoea.
A man in his 50s presented with abscessed aortic valve methicillin-sensitive Staphylococcus aureus endocarditis, received intravenous antibiotics and underwent bioprosthetic aortic valve replacement with removal of all infected tissues. He returned 18 days later with severe dyspnoea, subjective fever and bilateral lower extremity oedema. Physical examination revealed tachypnoea and tachycardia without fever, prominent neck CV waves visible at 90°, left parasternal heave, 3/6 holosystolic murmur across the precordium, lung rales and severe peripheral oedema. C reactive protein was 211 mg/L (normal <8 mg/L). Blood cultures were obtained. ECG showed sinus tachycardia and right axis deviation. Transthoracic echocardiogram (TTE) parasternal zoomed short-axis systolic frame (figure 1A, B), apical four-chamber systolic frame (figure 1C) and subcostal continuous wave (CW) Doppler (figure 1D), are shown. Given the clinical presentation and TTE findings, what is the diagnosis?Severe tricuspid regurgitation due to extension of endocarditisAortic valve obstruction resulting in severe pulmonary hypertensionAcquired Gerbode defectAortic valve dehiscence with severe paraprosthetic regurgitation.


Mass spectrometry analyses of rat 2b myosin heavy chain isoform.
We have separated 2b myosin heavy chain (MyHC) isoform from the rat extensor digitorum longus muscle by SDS-PAGE and analyzed it by two subsequent mass spectrometry techniques. After tryptic digestion, the obtained peptides were identified by Matrix-Assisted Laser Desorption/Ionisation reflectron Time of Flight mass spectrometry (MALDI-TOF MS) and sequenced by liquid chromatography tandem mass spectrometry (ESI LC/MS/MS). The analyzed peptides proportionally covered 30 % of the 2b MyHC isoform sequence. The results suggest that the primary structure is identical with the highest probability to a NCBI database record ref|NP_062198.1|, representing the last updated record of rat 2b isoform. Nonetheless, four peptides carrying amino acid substitution(s) in comparison with the NCBI database record were identified.


An overview of nitrate sources and operating processes in arid and semiarid aquifer systems.
Nitrate concentration in most aquifers in arid and semi-arid areas has increased in the past several decades as a result of human activities. Under the predominantly oxic conditions of these aquifers, denitrification is inhibited, allowing nitrate, a soluble and stable form of nitrogen (N), to accumulate. Because of its close association with municipal and agricultural wastes, nitrate is commonly used as an indicator of anthropogenic contamination. Aquifers affected by agricultural waste may contain salts from irrigation returns and herbicides in addition to nitrates. Preventing leakage from soil to deeper parts of the aquifer is thus a priority in the sustainable management of aquifers in arid and semiarid areas. Studies report a wide range of nitrate concentrations distributed non-uniformly within the aquifer, with roughly 40% and 20% of sampled wells exceeding 50mg/L nitrate in shallow and deep parts of the aquifer respectively. In aquifers at risk of becoming contaminated, nitrate isotopes (δ15N, δ18O, Δ17O) can be used to identify the source of nitrogen as mineral or organic fertilizer, sewage, or atmospheric deposition. A variety of mathematical models (crop, hydrological, geochemical, or a combination of them) have been successful in identifying best practices that minimize N leakage without negatively affecting crop yield. In addition, field research in crop management, e.g., conservation agriculture, has yielded promising results in determining the adequate dosage and time of application of fertilizers to reduce N losses. Examples of key dryland aquifers impacted by nitrate are discussed, and some of the most pressing challenges to achieve sustainability are presented.


Indoor black carbon and brown carbon concentrations from cooking and outdoor penetration: insights from the HOMEChem study.
Particle emissions from cooking are a major contributor to residential indoor air pollution and could also contribute to ambient concentrations. An important constituent of these emissions is light-absorbing carbon, including black carbon (BC) and brown carbon (BrC). This work characterizes the contributions of indoor and outdoor sources of BC and BrC to the indoor environment by concurrently measuring real-time concentrations of these air pollutants indoors and outdoors during the month-long HOMEChem study. The median indoor-to-outdoor ratios of BC and BrC during the periods of no activity inside the test house were 0.6 and 0.7, respectively. The absorption Ångström exponent was used to characterize light-absorbing particle emissions during different activities and ranged from 1.1 to 2.7 throughout the campaign, with the highest value (indicative of BrC-dominated emissions) observed during the preparation of a simulated Thanksgiving Day holiday style meal. An indoor BC exposure assessment shows that exposure for an occupant present in the kitchen area was ∼4 times higher during Thanksgiving Day experiments (primarily due to candle burning) when compared to the background conditions.


Excess triiodothyronine as a risk factor of coronary events.
Abnormalities in cardiac function, eg, arrhythmias and congestive heart failure, often accompany thyrotoxicosis. A relationship between thyroid hormone excess and the cardiac complications of angina pectoris and myocardial infarction (MI) remains largely speculative. The results of thyroid function studies on blood samples drawn from a total of 1049 patients (aged 40 years or older) immediately on emergency medical admission were related to frequencies of angina pectoris and myocardial infarction as determined according to current diagnostic algorithms. After 3 years, those patients who had initially presented with angina pectoris or acute MI were observed for subsequent coronary events; of these (n=185), 98% of the subjects (n=181) could be reevaluated. On hospital admission, the relative rate of angina pectoris and MI was markedly high (odds ratio, 2.6; 95% confidence interval, 1.3-5.2; P=.007) in patients with elevated serum free and total triiodothyronine (T(3)) levels. An initially elevated free T(3) level was a risk factor for subsequent coronary events during the 3-year follow-up (adjusted odds ratio, 4.8; 95% confidence interval, 1.3-17.4; P=.02). An elevation of serum free T(3) levels at hospital admission is associated with a 2.6-fold greater likelihood of the presence of a coronary event. Moreover, an initially elevated T(3) level is associated with a 3-fold higher risk of developing a subsequent coronary event during the next 3 years. Excess T(3) seemed to be a factor associated with the development and progression of acute myocardial ischemia.


Risk-preference differentiates orbitofrontal cortex responses to freely chosen reward outcomes.
To successfully evaluate potential courses of action and choose the most favorable, we must consider the outcomes that may result. Many choices involve risk, our assessment of which may lead us to success or failure in matters financial, legal or health-related. The orbitofrontal cortex (OFC) has been implicated as critical for evaluating choices based on risk. To measure how outcomes of risky decisions are represented in the OFC, we recorded the electrophysiological activity of single neurons while rats made behavioral responses to obtain rewards under conditions of either certainty or risk. Rats exhibited different risk-preferences when given the opportunity to choose. In risk-preferring rats, OFC responses were enhanced following the delivery of large rewards obtained under risk compared with smaller, certain rewards and reward omission. However, in risk-neutral rats, neurons showed similarly enhanced responses to both large rewards obtained under risk and smaller, certain rewards compared with reward omission. Thus, the responses of OFC neurons reflected the subjective evaluation of outcomes in individuals with different risk-preferences. Such enhanced neural responding to risky rewards may serve to bias individuals towards risk-preference in decision-making.


[Critical phenomena, phase equilibria, and the temperature and structural optimum of homeostasis, as revealed by a model system water-biopolymer-electrolyte].
Equations of spinodal and two quasispinodals corresponding to critical and supercritical phase transitions leading to a rise of different dynamic structures of solution in the phase diagram of a model system water-biopolymer-electrolyte were obtained. The section of the phase diagram was considered where there exists the probability of quasi-equilibrium monomer-cluster and the principle of water-ion homeostasis is realized. Based on these results, a possible mechanism of origination of unspecific adaptation reactions of a biomolecular system at the stage of chemical evolution was suggested.


[Primary diagnosis of syphilis by the ophthalmologist].
Throughout the developed world the reported incidence of sexually transmitted diseases is rising with previously unreported cases at rates of 40 - 70 %. According to German sources, recent epidemiological data show a considerable increase of syphilis especially among homosexual men in larger cities. We report on 4 patients (age 38 - 54) who were referred to our outpatient department because of vision loss of unknown origin. All were subsequently tested positive for syphilis. None of the patients (3 men, 1 woman) belonged to a risk group, only one described systemic symptoms (urethritis and arthritis). The ocular manifestations of syphilis were broad: granulomatous and non-granulomatous anterior uveitis, papillitis, and chorioretinitis. Two patients also tested positive for HIV. After systemic antibiotic therapy, the ocular diseases stabilized. Diagnosis and therapy of syphilis is cost-effective. The rise of syphilis especially in urban areas necessitates a high level of suspicion dealing with patients with intraocular inflammation of unknown origin. Lues serology should be incorporated into routine lab diagnostics to aid in the detection of such cases.


Diallel panel reveals a significant impact of low-frequency genetic variants on gene expression variation in yeast.
Unraveling the genetic sources of gene expression variation is essential to better understand the origins of phenotypic diversity in natural populations. Genome-wide association studies identified thousands of variants involved in gene expression variation, however, variants detected only explain part of the heritability. In fact, variants such as low-frequency and structural variants (SVs) are poorly captured in association studies. To assess the impact of these variants on gene expression variation, we explored a half-diallel panel composed of 323 hybrids originated from pairwise crosses of 26 natural Saccharomyces cerevisiae isolates. Using short- and long-read sequencing strategies, we established an exhaustive catalog of single nucleotide polymorphisms (SNPs) and SVs for this panel. Combining this dataset with the transcriptomes of all hybrids, we comprehensively mapped SNPs and SVs associated with gene expression variation. While SVs impact gene expression variation, SNPs exhibit a higher effect size with an overrepresentation of low-frequency variants compared to common ones. These results reinforce the importance of dissecting the heritability of complex traits with a comprehensive catalog of genetic variants at the population level.


Are Psychology Journals Anti-replication? A Snapshot of Editorial Practices.
Recent research in psychology has highlighted a number of replication problems in the discipline, with publication bias - the preference for publishing original and positive results, and a resistance to publishing negative results and replications- identified as one reason for replication failure. However, little empirical research exists to demonstrate that journals explicitly refuse to publish replications. We reviewed the instructions to authors and the published aims of 1151 psychology journals and examined whether they indicated that replications were permitted and accepted. We also examined whether journal practices differed across branches of the discipline, and whether editorial practices differed between low and high impact journals. Thirty three journals (3%) stated in their aims or instructions to authors that they accepted replications. There was no difference between high and low impact journals. The implications of these findings for psychology are discussed.


Hypoglycemic and antidiabetic effect of ethanolic extract of leaves of Annona squamosa L. in experimental animals.
The ethanolic extract of Annona squamosa L. (Annonaceae) leaves was administered orally at different doses to normal as well as streptozotocin (STZ)-induced diabetic rats and alloxan-induced diabetic rabbits. The dose of 350 mg/kg body weight (bw) reduced the fasting blood glucose (FBG) level by 6.0% within 1 h, whereas, the peak blood glucose at 1 h during glucose tolerance test (GTT) was reduced by 17.1% in normal rats. The same dose of ethanolic extract reduced FBG by 26.8% and improved glucose tolerance by 38.5 and 40.6% at 1 and 2 h, respectively, during GTT in alloxan-induced diabetic rabbits. In STZ-diabetic rats, a fall of 13.0% in FBG and an improvement in glucose tolerance by 37.2 and 60.6% at 1 and 2 h, respectively, was observed during GTT. The dose of 350 mg/kg bw of ethanolic extract in 10-day treatment of a group of STZ-diabetic rats produced 73.3% fall in FBG level and no sugar was observed in fasting urine. Treatment of severely-diabetic rabbits for 15 days with a dose of 350 mg/kg of extract reduce FBG by 52.7% and urine sugar by 75%. It brought about fall in the level of total cholesterol (TC) by 49.3% with increase of 30.3% in high-density lipoprotein (HDL) and decrease of 71.9 and 28.7% in low-density lipoprotein (LDL) and triglycerides (TG) levels, respectively.


Ertapenem.
Given their broad-spectrum and safety, carbapenems are a widely used class of antibiotics, especially in the treatment of hospital-acquired infections including infections due to multidrug-resistant organisms. Ertapenem is a unique member of this class, with a narrower spectrum that lacks reliable activity against Pseudomonas and Enterococcus. Given its spectrum and half-life of 4 h it is better suited to use in community-acquired infections and it is particularly well positioned for use in the outpatient setting. Chemistry, mechanism of action, pharmacokinetics/pharmacodynamics, safety and indications for use will be covered in this review. Similar to other beta-lactams, the carbapenems inhibit cell wall synthesis by binding to and inhibiting penicillin-binding proteins. Their resistance to beta-lactamases including AmpC and extended-spectrum beta-lactamases enhances their usefulness. Similar to other beta-lactams, ertapenem exhibits time-dependent killing. Given this profile, ertapenem has been found to be useful in intra-abdominal infections, acute pelvic infections, complicated skin and skin structure infections, community-acquired pneumonia and complicated urinary tract infections. This review will enable the reader to understand differences between the different carbapenems, especially with regard to ertapenem. Once an understanding is gained with regard to pharmacology and microbiology, the reader will be positioned to understand better those circumstances in which use of ertapenem should be considered. Substantial differences between the carbapenems exist. Ertapenem has unique characteristics that may make it useful in specific clinical circumstances that are detailed in this review.


Early development, stress and depression across the life course: pathways to depression in a national British birth cohort.
The aetiology of depression is multifactorial, with biological, cognitive and environmental factors across the life course influencing risk of a depressive episode. There is inconsistent evidence linking early life development and later depression. The aim of this study was to investigate relationships between low birthweight (LBW), infant neurodevelopment, and acute and chronic stress as components in pathways to depression in adulthood. The sample included 4627 members of the National Survey of Health and Development (NSHD; the 1946 British birth cohort). Weight at birth, age of developmental milestones, economic deprivation in early childhood, acute stressors in childhood and adulthood, and socio-economic status (SES) in adulthood were assessed for their direct and indirect effects on adolescent (ages 13 and 15 years) and adult (ages 36, 43 and 53 years) measures of depressive symptoms in a structural equation modelling (SEM) framework. A structural equation model developed to incorporate all variables exhibited excellent model fit according to several indices. The path of prediction from birthweight to age of developmental milestones to adolescent depression/anxiety to adult depression/anxiety was significant (p < 0.001). Notably, direct paths from birthweight (p = 0.25) and age of developmental milestones (p = 0.23) to adult depression were not significant. Childhood deprivation and stressors had important direct and indirect effects on depression. Stressors in adulthood were strongly associated with adult depression. Depression in adulthood is influenced by an accumulation of stressors across the life course, including many that originate in the first years of life. Effects of early-life development on mental health appear by adolescence.


Bursts and the efficacy of selective serotonin reuptake inhibitors.
We present a new hypothesis for the efficacy of selective serotonin reuptake inhibitors (SSRIs). We propose that SSRIs bring the response to the phasic firing of raphe nucleus cells back to normal, even though the average extracellular 5HT concentration remains low. We discuss burst firing in the raphe nuclei and use mathematical models to argue that tonic firing and phasic firing may be decoupled and may come from different mechanisms. We use a mathematical model for serotonin synthesis, release, and reuptake in terminals to illustrate the responses in terminal regions to bursts in a normal individual and in an individual with low vesicular serotonin. We then show that acute doses of SSRIs do not bring the response to bursts back to normal, but that chronic doses do return the response to normal. These model results need to be confirmed by new electrophysiological and pharmacological experiments.


Survivin expression correlates with apoptosis resistance after lymphocyte activation and is found preferentially in memory T cells.
The prevention of apoptosis may be critical for immunological function. Survivin is a recently cloned member of the inhibitor of apoptosis protein family. We analyzed survivin expression before and after lymphocyte activation in isolated cell populations. Prior to activation, survivin was undetectable. After activation with IL-2 and OKT-3, CD3(+) cells expressed survivin. Next, we correlated survivin expression with Fas, FasL and the amount of apoptosis over time in culture. After activation, survivin was readily detected by day 2 and decreased thereafter. Prior to activation (day 0), Fas was present on 60% of the cells and on 100% by days 2-6. Peak FasL mRNA expression was at day 2. During peak survivin expression (days 2-4) the apoptotic fraction was low, but when survivin expression decreased the apoptotic fraction increased rapidly. Finally, we found that CD45RO(+) memory T cells showed a higher expression of survivin than did CD45RA(+) naive T cells after activation. These results suggest that survivin may contribute to T-cell survival early in T-cell responses as well as in memory immune responses.


Evaluation of loxapine hydrochloride oral concentrate (loxitane C) in acute schizophrenia.
In an open study of 4-weeks duration, both an oral liquid concentrate formulation of loxapine hydrochloride (LOXITANE C) and capsules of loxapine succinate (LOXITANE) were administered to 11 acutely disturbed schizophrenic patients. Optimal dosage levels achieved with the concentrate proved satisfactory with the capsules. Efficacy evaluation with the Brief Psychiatric Rating Scale (BPRS), Systematic Nurses' Observation of Psychopathology (SNOOP), and Clinical Global Impressions (CGI) indicated rapid improvement with concentrate administration and continued improvement with capsule administration. All but two side effects were extrapyramidal or sedative, all but one were mild or moderate in severity, and the frequency was similar with the two formulations. Cardiovascular and clinical laboratory findings remained essentially unchanged with both formulations.


Immunotherapy with a HER2-Targeting Listeria Induces HER2-Specific Immunity and Demonstrates Potential Therapeutic Effects in a Phase I Trial in Canine Osteosarcoma.
Recombinant Listeria vaccines induce tumor-specific T-cell responses that eliminate established tumors and prevent metastatic disease in murine cancer models. We used dogs with HER2/neu(+) appendicular osteosarcoma, a well-recognized spontaneous model for pediatric osteosarcoma, to determine whether a highly attenuated, recombinant Listeria monocytogenes expressing a chimeric human HER2/neu fusion protein (ADXS31-164) could safely induce HER2/neu-specific immunity and prevent metastatic disease. Eighteen dogs that underwent limb amputation or salvage surgery and adjuvant chemotherapy were enrolled in a phase I dose escalation clinical trial and received either 2 × 10(8), 5 × 10(8), 1 × 10(9), or 3.3 × 10(9) CFU of ADXS31-164 intravenously every 3 weeks for 3 administrations. Only low-grade, transient toxicities were observed. ADXS31-164 broke peripheral tolerance and induced antigen-specific IFNγ responses against the intracellular domain of HER2/neu in 15 of 18 dogs within 6 months of treatment. Furthermore, ADXS31-164 reduced the incidence of metastatic disease and significantly increased duration of survival time and 1-, 2-, and 3-year survival rates when compared with a historical control group with HER2/neu(+) appendicular osteosarcoma treated with amputation and chemotherapy alone. These findings demonstrate that ADXS31-164 administered in the setting of minimal residual disease can induce HER2/neu-specific immunity and may reduce the incidence of metastatic disease and prolong overall survival in a clinically relevant, spontaneous, large animal model of cancer. These findings, therefore, have important translational relevance for children with osteosarcoma and adults with other HER2/neu(+) cancers. Clin Cancer Res; 22(17); 4380-90. ©2016 AACR.


[Coping with stress as a paradigm of the psychopathology in childhood and adolescence].
Stress coping can be valid for a paradigm of psychopathology of children and adolescents. This concept is supported by an extensive literature which shows the way of coping from a "footnote to stress theory" (Lazarus) to its central concept. Neither the "objective" stress (stimulus side) nor the specificity of stress reaction (adaptation syndrome) are the important factors, but rather the mediation between stress coping and the individual, the transaction in the sense of an exchange process. Above all, permanent personality attributes have been examined here, most intensively extraversion/introversion and neuroticism, and recently, more and more often coping processes. The research situation also confirms in its approach the concept of Lazarus (1981) in the field of child and adolescent psychiatry. According to Lazarus, stressors are less meaningful as a cause of psychiatric disturbances than cognitive-emotional reaction mediating coping processes. They should be examined preferably under natural conditions; a combination of standard research strategies with individual case analyses is recommended. Therapeutically, it is essential to help children and adolescents to achieve an adequate correlation between their own capabilities and limitations on the one hand, and of conflicts and stress situations on the other hand, whereby the adolescent (young) patient is interested in a quick solution to personal and interpersonal crises. He is not at all interested in its extension in the sense of a 'Sturm-und-Drang'-idea of adolescent age. Hence coping cannot be confined only to coping with a functional crisis; it must rather aspire to achieve a schematisation of the situation in accordance with the truth.


Response of the Saccharomyces cerevisiae Mpk1 mitogen-activated protein kinase pathway to increases in internal turgor pressure caused by loss of Ppz protein phosphatases.
The Mpk1 pathway of Saccharomyces cerevisiae is a key determinant of cell wall integrity. A genetic link between the Mpk1 kinase and the Ppz phosphatases has been reported, but the nature of this connection was unclear. Recently, the Ppz phosphatases were shown to be regulators of K+ and pH homeostasis. Here, we demonstrate that Ppz-deficient strains display increased steady-state K+ levels and sensitivity to increased KCl concentrations. Given these observations and the fact that K+ is the major determinant of intracellular turgor pressure, we reasoned that the connection between PPZ1 and -2 and MPK1 was due to the combination of increased internal turgor pressure in Ppz-deficient strains and cell wall instability observed in strains lacking MPK1. Accordingly, the MPK1 gene was up-regulated, the Mpk1 protein was overexpressed, and the phosphorylated active form was more abundant in Ppz-deficient strains. Moreover, the expression of genes previously identified as targets of the Mpk1 pathway are also up-regulated in strains lacking PPZ1 and -2. The transcriptional and posttranslational modifications of Mpk1 were not observed when the internal K+ concentration (and thus turgor pressure) was lowered by disrupting the TRK1 and -2 K+ transporter genes or when the cell wall was stabilized by the addition of sorbitol. Moreover, we present genetic evidence showing that both the Wsc1 and Mid2 branches of the Mpk1 pathway contribute to this response. Finally, despite its role in G1/S transition, increased levels of activated Mpk1 do not appear to be responsible for the cell cycle phenotype observed in Ppz-deficient strains.


Channeling through Two Stacked Guanine Quartets of One and Two Alkali Cations in the Li+, Na+, K+, and Rb+ Series. Assessment of the Accuracy of the SIBFA Anisotropic Polarizable Molecular Mechanics Potential.
Stacking of guanine quartets (GQs) can trigger the formation of DNA or RNA quadruple helices, which play numerous biochemical roles. The GQs are stabilized by alkali cations, mainly K+ and Na+, which can reside in, or channel through, the central axis of the GQ stems. Further, ion conduction through GQ wires can be leveraged for nanochemistry applications. G-quadruplex systems have been extensively studied by classical molecular dynamics (MD) simulations using pair-additive force fields or by quantum-chemical (QC) calculations. However, the non-polarizable force fields are very approximate, while QC calculations lack the necessary sampling. Thus, ultimate description of GQ systems would require long-enough simulations using advanced polarizable molecular mechanics (MM). However, to perform such calculations, it is first mandatory to evaluate the method's accuracy using benchmark QC. We report such an evaluation for SIBFA polarizable MM, bearing on the channeling (movement) of an alkali cation (Li+, Na+, K+, or Rb+) along the axis of two stacked G quartets interacting with either one or two ions. The QC energy profiles display markedly different features depending upon the cation but can be retrieved in the majority of cases by the SIBFA profiles. An appropriate balance of first-order (electrostatic and short-range repulsion) and second-order (polarization, charge-transfer, and dispersion) contributions within ΔE is mandatory. With two cations in the channel, the relative weights of the second-order contributions increase steadily upon increasing the ion size. In the G8 complexes with two K+ or two Rb+ cations, the sum of polarization and charge-transfer exceeds the first order terms for all ion positions.


Smad3 promotes alkaline phosphatase activity and mineralization of osteoblastic MC3T3-E1 cells.
Transforming growth factor (TGF) beta is abundantly stored in bone matrix and appears to regulate bone metabolism. Although the Smad family proteins are critical components of the TGF-beta signaling pathways, the roles of Smad3 in the expression of osteoblastic phenotypes remain poorly understood. Therefore, this study was performed to clarify the roles of Smad3 in the regulation of proliferation, expression of bone matrix proteins, and mineralization in osteoblasts by using mouse osteoblastic cell line MC3T3-E1 cells stably transfected with Smad3. Smad3 significantly inhibited [3H]thymidine incorporation and fluorescent intensity of the MTT-dye assay, compared with empty vector. Moreover, Smad3 increased the levels of type I procollagen, osteopontin (OPN), and matrix Gla protein (MGP) mRNA in Northern blotting. These effects of Smad3 mimicked the effects of TGF-beta on the same cells. On the other hand, Smad3 greatly enhanced ALP activity and mineralization of MC3T3-E1 cells compared with empty vector, although TGF-beta inhibited ALP activity and mineralization of wild-type MC3T3-E1 cells. A type I collagen synthesis inhibitor L-azetidine-2-carboxylic acid, as well as osteocalcin (OCN), significantly antagonized Smad3-stimulated ALP activity and mineralization of MC3T3-E1 cells. In conclusion, this study showed that in mouse osteoblastic cells, Smad3 inhibited proliferation, but it also enhanced ALP activity, mineralization, and the levels of bone matrix proteins such as type I collagen (COLI), OPN, and MGP. We propose that Smad3 plays an important role in osteoblastic bone formation and might help to elucidate the transcriptional mechanism of bone formation and possibly lead to the development of bone-forming drugs.


Acroangiodermatitis (pseudo-Kaposi sarcoma): a rarely-recognized condition. A case on the plantar aspect of the foot associated with chronic venous insufficiency.
Acroangiodermatitis, often known as pseudo-Kaposi sarcoma, is an uncommon angioproliferative entity related to chronic venous insufficiency, arteriovenous fistulae, paralysed limbs, amputation stumps, vascular syndromes and conditions associated with thrombosis. It presents most frequently as purple macules, papules or plaques in the dorsal aspects of the feet, especially the toes, and the malleoli. We report a case of acroangiodermatitis in the plantar aspect of the foot, misdiagnosed for two years, in which haematoxylin-eosin hystopathological stain and immunolabeling with CD34 histochemistry examination were decisive for diagnosis. Patient had chronic venous insufficiency. The lesion responded well to the treatment with a combination of leg elevation and compression.


Constitution of the twin polymerase of DNA polymerase III holoenzyme.
It is speculated that DNA polymerases which duplicate chromosomes are dimeric to provide concurrent replication of both leading and lagging strands. DNA polymerase III holoenzyme (holoenzyme), is the 10-subunit replicase of the Escherichia coli chromosome. A complex of the alpha (DNA polymerase) and epsilon (3'-5' exonuclease) subunits of the holoenzyme contains only one of each protein. Presumably, one of the eight other subunit(s) functions to dimerize the alpha epsilon polymerase within the holoenzyme. Based on dimeric subassemblies of the holoenzyme, two subunits have been elected as possible agents of polymerase dimerization, one of which is the tau subunit (McHenry, C. S. (1982) J. Biol. Chem. 257, 2657-2663). Here, we have used pure alpha, epsilon, and tau subunits in binding studies to determine whether tau can dimerize the polymerase. We find tau binds directly to alpha. Whereas alpha is monomeric, tau is a dimer in its native state and thereby serves as an efficient scaffold to dimerize the polymerase. The epsilon subunit does not associate directly with tau but becomes dimerized in the alpha epsilon tau complex by virtue of its interaction with alpha. We have analyzed the dimeric alpha epsilon tau complex by different physical methods to increase the confidence that this complex truly contains a dimeric polymerase. The tau subunit is comprised of the NH2-terminal two-thirds of tau but does not bind to alpha epsilon, identifying the COOH-terminal region of tau as essential to its polymerase dimerization function. The significance of these results with respect to the organization of subunits within the holoenzyme is discussed.


Predicting diabetic nephropathy using a multifactorial genetic model.
The tendency to develop diabetic nephropathy is, in part, genetically determined, however this genetic risk is largely undefined. In this proof-of-concept study, we tested the hypothesis that combined analysis of multiple genetic variants can improve prediction. Based on previous reports, we selected 27 SNPs in 15 genes from metabolic pathways involved in the pathogenesis of diabetic nephropathy and genotyped them in 1274 Ashkenazi or Sephardic Jewish patients with Type 1 or Type 2 diabetes of >10 years duration. A logistic regression model was built using a backward selection algorithm and SNPs nominally associated with nephropathy in our population. The model was validated by using random "training" (75%) and "test" (25%) subgroups of the original population and by applying the model to an independent dataset of 848 Ashkenazi patients. The logistic model based on 5 SNPs in 5 genes (HSPG2, NOS3, ADIPOR2, AGER, and CCL5) and 5 conventional variables (age, sex, ethnicity, diabetes type and duration), and allowing for all possible two-way interactions, predicted nephropathy in our initial population (C-statistic = 0.672) better than a model based on conventional variables only (C = 0.569). In the independent replication dataset, although the C-statistic of the genetic model decreased (0.576), it remained highly associated with diabetic nephropathy (χ(2) = 17.79, p<0.0001). In the replication dataset, the model based on conventional variables only was not associated with nephropathy (χ(2) = 3.2673, p = 0.07). In this proof-of-concept study, we developed and validated a genetic model in the Ashkenazi/Sephardic population predicting nephropathy more effectively than a similarly constructed non-genetic model. Further testing is required to determine if this modeling approach, using an optimally selected panel of genetic markers, can provide clinically useful prediction and if generic models can be developed for use across multiple ethnic groups or if population-specific models are required.


Intrathymic adeno-associated virus gene transfer rapidly restores thymic function and long-term persistence of gene-corrected T cells.
Patients with T-cell immunodeficiencies are generally treated with allogeneic hematopoietic stem cell transplantation, but alternatives are needed for patients without matched donors. An innovative intrathymic gene therapy approach that directly targets the thymus might improve outcomes. We sought to determine the efficacy of intrathymic adeno-associated virus (AAV) serotypes to transduce thymocyte subsets and correct the T-cell immunodeficiency in a zeta-associated protein of 70 kDa (ZAP-70)-deficient murine model. AAV serotypes were injected intrathymically into wild-type mice, and gene transfer efficiency was monitored. ZAP-70-/- mice were intrathymically injected with an AAV8 vector harboring the ZAP70 gene. Thymus structure, immunophenotyping, T-cell receptor clonotypes, T-cell function, immune responses to transgenes and autoantibodies, vector copy number, and integration were evaluated. AAV8, AAV9, and AAV10 serotypes all transduced thymocyte subsets after in situ gene transfer, with transduction of up to 5% of cells. Intrathymic injection of an AAV8-ZAP-70 vector into ZAP-70-/- mice resulted in a rapid thymocyte differentiation associated with the development of a thymic medulla. Strikingly, medullary thymic epithelial cells expressing the autoimmune regulator were detected within 10 days of gene transfer, correlating with the presence of functional effector and regulatory T-cell subsets with diverse T-cell receptor clonotypes in the periphery. Although thymocyte reconstitution was transient, gene-corrected peripheral T cells harboring approximately 1 AAV genome per cell persisted for more than 40 weeks, and AAV vector integration was detected. Intrathymic AAV-transduced progenitors promote a rapid restoration of the thymic architecture, with a single wave of thymopoiesis generating long-term peripheral T-cell function.


[An immunohistochemical study of placenta-specific alpha 2-microglobulin in human ontogeny].
Tissues of adult man and fetus were studied by indirect immunoperoxidase technique using affinity-purified antibodies against placenta-specific alpha 2-microglobulin (APM-2). APM-2 proved to be synthesized mucous membrane cells of uterus during secretory phase of the menstrual cycle. APM-2 was also found in atretic follicle of fertile woman ovary as well as in I order oocytes from the fetus ovary (26 weeks). In men APM-2 is synthesized in seminal vesicles. APM-2 was found both in cytotrophoblast and in syncytiotrophoblast as shown by study of chorionic villi.


A chemogenomics based approach for deorphanization of testicular receptor 4: An orphan receptor of nuclear receptor superfamily.
Orphan Receptor of Nuclear Receptor superfamily is the one with no known endogenous ligands. Many of these orphan receptors are associated with different types of diseases and therefore deserve special attention to find the potential ligands they would be associated with. The major task of molecular pharmacology is the deorphanization of the large number of nuclear receptors with unidentified endogenous agonists. The deorphanization provides a promising research for new therapeutics. The Testicular Receptor 4 being negative modulator to other members of the nuclear receptor superfamily, is one of the Orphan members of this family and is associated with prostate cancer, breast cancer, sickle cell anemia and joint diseases. The knowledge that related receptors of the same family often have ligands with similar structural features has helped us to utilize the chemogenomic approach to deorphanize the orphan receptor. Chemogenomics approach involves screening of known ligands of a protein family having analogous domain architecture for identification of new leads for existing protein family members. The deorphanization involved the database homology searching, followed by domain identification, active site prediction, sequence and structure comparative studies. A ligand library set was prepared based on these studies and was used to deorphanize the receptor. The molecular docking study conducted using PyRx revealed that estradiol and tretinion as a potential ligand for Testicular Receptor 4.


Evidence for involvement of Saccharomyces cerevisiae protein kinase C in glucose induction of HXT genes and derepression of SUC2.
The PKC1 gene in the yeast Saccharomyces cerevisiae encodes protein kinase C that is known to control a mitogen-activated protein (MAP) kinase cascade consisting of Bck1, Mkk1 and Mkk2, and Mpk1. This cascade affects the cell wall integrity but the phenotype of Pkc1 mutants suggests additional targets which have not yet been identified. We show that a pkc1Delta mutant, as opposed to mutants in the MAP kinase cascade, displays two major defects in the control of carbon metabolism. It shows a delay in the initiation of fermentation upon addition of glucose and a defect in derepression of SUC2 gene after exhaustion of glucose from the medium. After addition of glucose the production of both ethanol and glycerol started very slowly. The V(max) of glucose transport dropped considerably and Northern blot analysis showed that induction of the HXT1, HXT2 and HXT4 genes was strongly reduced. Growth of the pkc1Delta mutant was absent on glycerol and poor on galactose and raffinose. Oxygen uptake was barely present. Derepression of invertase activity and SUC2 transcription upon transfer of cells from glucose to raffinose was deficient in the pkc1Delta mutant as opposed to the wild-type. Our results suggest an involvement of Pkc1p in the control of carbon metabolism which is not shared by the downstream MAP kinase cascade.


Quinine-induced thrombocytopenia: drug-dependent GPIb/IX antibodies inhibit megakaryocyte and proplatelet production in vitro.
The development of immune cytopenias is a well-recognized side effect of many drugs. Quinine- and quinidine-dependent antibodies are classic examples of drug-induced effects that cause severe, life-threatening thrombocytopenia. Whereas the effects of drug-dependent antibodies on platelets have been well documented, their effects on megakaryocyte (Mk) biology are still unclear. We analyzed sera from several quinine-induced thrombocytopenia (QITP) patients on highly pure Mks (98% glycoprotein IIb-positive [GPIIb(+)]; 92% GPIX(+)) derived from human CD34(+) cells cultured with human thrombopoietin. We demonstrate by flow cytometry and confocal microscopy that QITP IgGs bind Mks efficiently in the presence of quinine. Incubation of day-4 Mks with QITP sera or purified IgG resulted in induction of apoptosis, a significant decrease in cell viability, and an increase in cell death. Furthermore, QITP sera preferentially reduced the number of late GPIX(+)/GPIbα(+) Mks and the number of receptors per cell in the surviving population. Ploidy distribution, lobularity, and average cell size of Mks remained unchanged after treatment. In addition, treated Mks showed a marked decrease in their proplatelet production capacity, suggesting that drug-dependent antibodies hinder platelet production. Therefore, QITP antibodies considerably reduce the proplatelet production capabilities of Mks despite undetectable effects on DNA content, morphology, and cell size.


Treatment with beta-adrenoceptor blockers reduces plasma melatonin concentration.
In treated hypertensive patients plasma melatonin levels were lower in subjects receiving beta-adrenoceptor blockers than those treated with diuretics. Melatonin concentrations in middle-aged and young control subjects were similar to each other and to those of the diuretic-treated patients. The results suggest that treatment with beta-adrenoceptor blockers causes a persistent reduction in plasma melatonin but it is unclear if this finding has clinical implications.


Naturally occurring autoantibodies against beta-amyloid: investigating their role in transgenic animal and in vitro models of Alzheimer's disease.
Alzheimer's disease (AD) is a neurodegenerative disorder primarily affecting regions of the brain responsible for higher cognitive functions. Immunization against β-amyloid (Aβ) in animal models of AD has been shown to be effective on the molecular level but also on the behavioral level. Recently, we reported naturally occurring autoantibodies against Aβ (NAbs-Aβ) being reduced in Alzheimer's disease patients. Here, we further investigated their physiological role: in epitope mapping studies, NAbs-Aβ recognized the mid-/C-terminal end of Aβ and preferentially bound to oligomers but failed to bind to monomers/fibrils. NAbs-Aβ were able to interfere with Aβ peptide toxicity, but NAbs-Aβ did not readily clear senile plaques although early fleecy-like plaques were reduced. Administration of NAbs-Aβ in transgenic mice improved the object location memory significantly, almost reaching performance levels of wild-type control mice. These findings suggest a novel physiological mechanism involving NAbs-Aβ to dispose of proteins or peptides that are prone to forming toxic aggregates.


On the origin of the transthyretin Val30Met familial amyloid polyneuropathy.
Transthyretin (TTR) familial amyloid polyneuropathy is a severe autosomal dominant neuropathy of adulthood, frequently linked to the pathogenic Val30Met variant of the TTR gene. The condition was initially described in northern Portugal, which is the first focus of the disease. Other important clusters of families are found in Sweden, Japan and South America. The origin of the Val30Met mutation and its distribution through the populations remains unclear. In the present work, we aimed at refining the history of the Val30Met mutation in patients affected with TTR amyloid neuropathy from Portugal, Sweden and Brazil. The decay of haplotype sharing was studied in 60 patients to estimate the age of the Most Recent Common Ancestor (MRCA) of mutation carriers in these populations. Our results showed a common haplotype in Portuguese and Brazilian patients and an age estimate of the MRCA of 750 and 650 years, respectively. In contrast, a different haplotype was found in the Swedish Val30Met patients with a corresponding age estimate for the MRCA, of 375 years. This work strengthens the hypothesis of different founders in Portuguese and Swedish Val30Met carriers and suggested a Portuguese origin of the Brazilian mutation. The age estimates of the MRCA are in line with the current historical knowledge of these populations.


Tracheal smooth muscle.
Contraction of tracheal smooth muscle requires the binding of Ca2+ to calmodulin, which then binds to and activates MLCK. The Ca2+-calmodulin-MLCK complex catalyzes the phosphorylation of myosin, which causes contraction by stimulating actin-activated Mg2+-ATPase activity of myosin. Myosin phosphorylation appears to be a transient event that is responsible for a high velocity of shortening. The mechanism responsible for maintenance of isometric force is unknown, although a second Ca2+-dependent mechanism with a greater sensitivity to Ca2+ than the activation of MLCK has been hypothesized. Force would be maintained through the slow cycling of nonphosphorylated cross-bridges or a small population of phosphorylated cross-bridges. Tracheal smooth muscle utilizes both extracellular and intracellular pools of Ca2+ for contraction. Moreover, the membrane channels through which extracellular Ca2+ passes have been subdivided into potential-dependent channels (PDCs) and receptor-operated channels (ROCs) independent of membrane potential. The relative extent to which extracellular and intracellular sources of Ca2+ as well as PDCs and ROCs are utilized depends on the agonist used for contraction, its concentration, and the type and location of the smooth muscle being investigated. Calcium antagonists such as verapamil and nifedipine, which reportedly block PDCs but not ROCs, are much better inhibitors of tracheal smooth muscle contractions induced by serotonin than those induced by acetylcholine, histamine, and leukotriene D4, indicating an effect of these latter three agents on ROCs. Relaxation of tracheal smooth muscle following stimulation of beta-adrenergic receptors most likely results from an increase in cAMP that stimulates a cAMP-dependent protein kinase to catalyze a protein phosphorylation that leads to relaxation by decreasing the intracellular concentration of Ca2+. The primary mechanisms whereby cAMP is thought to reduce intracellular Ca2+ to effect relaxation include: activation of a calmodulin-sensitive Ca2+ ATPase in the plasma and sarcoplasmic reticulum membranes, and extrusion of Ca2+ by a Na+-Ca2+ exchange mechanism coupled to Na+-K+-ATPase in the cell membrane. A more controversial mechanism for relaxation that bypasses Ca2+ might involve the dephosphorylation of myosin. Leukotrienes are released by various stimuli, including immunologic challenge, and have been considered as important mediators of bronchoconstriction in allergic asthma.(ABSTRACT TRUNCATED AT 400 WORDS)


Characterization and tissue-specific expression of human GSK-3-binding proteins FRAT1 and FRAT2.
We have isolated the entire coding sequence of human FRAT2 (frequently rearranged in advanced T-cell lymphomas-2). It exhibits appreciable amino acid identity to FRAT1 (77%) which was initially isolated as frequently being overexpressed in a murine leukemia virus insertion model in murine tumors. FRAT proteins are thought to play a role in Wnt signaling. They can bind to glycogen synthase kinase-3 (GSK-3) and Dishevelled, two proteins involved in Wnt signal transduction. Both hFRAT1 and hFRAT2 are intronless genes localized to the same portion of chromosome 10q24.1 and separated by only 10.7 kb. In a broad range of human tissues FRAT1 and FRAT2 are readily detected and expressed in a near identical pattern. Both species are repressed when the human embryonal carcinoma cell line, NT2/D1, is induced to differentiate with all-trans retinoic acid (RA). This treatment had no appreciable effect on FRAT levels in two other RA-sensitive cell lines that were not of germ cell tumor origin. The overlapping expression patterns suggest these two genes share a regulatory region. Both FRAT genes exhibited three species of mRNA, which varied in representation between tissues. When transiently overexpressed in COS-1 cells, the FRAT proteins were detected in the cytosol and concentrated in the nucleus. Both hFRAT1 and hFRAT2 are implicated in the selective modulation of GSK-3 activity via the Wnt signaling pathway. This study provides a foundation from which to examine the role these proteins play in Wnt-dependent and -independent processes.


[Being a supervisor demands other skills than being a doctor].
Engagement with being a doctor is not an automatic qualification for being a good supervisor. But engagement with being a teacher is linked to a better evaluation as supervisor by trainees. Different personality traits are related to these different roles. Engagement is not only influenced by personality traits, however. The work environment can have a positive effect on engagement as well. If there are proper job resources that support the teacher's role, engagement with being a teacher will increase. In addition to proper teaching resources, supervisors also need training. It is important that supervisors get used to lifelong learning, just as doctors have to. Teaching competencies should be part of medical training. This might help to shape a working environment that is well equipped for teaching.


Defective oligodendrocyte development and severe hypomyelination in PDGF-A knockout mice.
There is a class of oligodendrocyte progenitors, called O-2A progenitors, that is characterized by expression of platelet-derived growth factor &agr;-receptors (PDGFR(&agr;)). It is not known whether all oligodendrocytes are derived from these PDGFRalpha-progenitors or whether a subset(s) of oligodendrocytes develops from a different, PDGFR alpha-negative lineage(s). We investigated the relationship between PDGF and oligodendrogenesis by examining mice that lack either PDGF-A or PDGF-B. PDGF-A null mice had many fewer PDGFR alpha-progenitors than either wild-type or PDGF-B null mice, demonstrating that proliferation of these cells relies heavily (though not exclusively) on PDGF-AA homodimers. PDGF-A-deficient mice also had reduced numbers of oligodendrocytes and a dysmyelinating phenotype (tremor). Not all parts of the central nervous system (CNS) were equally affected in the knockout. For example, there were profound reductions in the numbers of PDGFR alpha-progenitors and oligodendrocytes in the spinal cord and cerebellum, but less severe reductions of both cell types in the medulla. This correlation suggests a close link between PDGFRalpha-progenitors and oligodendrogenesis in most or all parts of the CNS. We also provide evidence that myelin proteolipid protein (PLP/DM-20)-positive cells in the late embryonic brainstem are non-dividing cells, presumably immature oligodendrocytes, and not proliferating precursors.


The pharmacokinetics of a single oral dose of mycophenolate mofetil in patients with varying degrees of renal function.
The purpose of this study was to determine the effect of renal function on the elimination and disposition of mycophenolic acid and its glucuronide metabolite (MPAG) after oral administration of the pro-drug mycophenolate mofetil. In addition, this study sought to examine hemodialysis removal of mycophenolic acid and its MPAG. Subjects were stratified into five groups on the basis of iohexol clearance. After an overnight fast, all subjects received a single 1 gm dose of mycophenolate mofetil. Plasma concentrations of mycophenolic acid and MPAG were measured from 0 to 96 hours after administration. Mycophenolic acid and MPAG maximum plasma concentration (Cmax) and the time to reach Cmax (tmax) for each group were determined from the mean plasma concentration-time profiles. Area under the plasma concentration-time curve values for mycophenolic acid and MPAG were calculated by the trapezoidal rule. The half-lives of mycophenolic acid and MPAG were calculated from the terminal portions of the concentration-time profiles. Mycophenolic acid clearance was not associated with changes in glomerular filtration rate (GFR). Cmax tended to increase as GFR declined. MPAG clearance correlated well with GFR (r2 = 0.905). Clearance of mycophenolic acid and MPAG were unaffected by hemodialysis. Clearance of mycophenolic acid after a single 1 gm oral dose of mycophenolate mofetil is unaffected by renal function. Clearance of mycophenolic acid is unaffected by hemodialysis. Diminished renal function should not require preemptive adjustment of 1 gm doses of mycophenolate mofetil; however dosage adjustment may be warranted on the basis of adverse effects or toxicity in individual patients. Mycophenolate mofetil can be administered irrespective of hemodialysis session without effect on mycophenolic acid exposure.


A descriptive epidemiology of lifetime trauma and the physical health status of older adults.
Three issues are evaluated in this study. The 1st involves examining the relationship between exposure to trauma over the life course and physical health status in old age. The 2nd has to do with seeing whether the relationship between trauma and health varies across 3 cohorts of older adults: the young-old (ages 65-74), the old-old (ages 75-84), and the oldest old (age 85 and over). The 3rd issue involves seeing whether the age at which a trauma was encountered is related to health in late life. Data from a nationwide survey of older people (N=1,518) reveal that trauma is associated with worse health. Moreover, the young-old appear to be at greatest risk. Finally, data suggest that trauma arising between the ages of 18 and 30 years, as well as ages 31 to 64 years, has the strongest relationship with current health.


Venous invasion may predict lymph node metastasis in early rectal cancer.
The aim of this study was to evaluate the role of histopathological and demographic characteristics in predicting lymph node metastasis in patients with adenocarcinoma of the rectum confined to the mucosal and submucosal layers. Fifty-nine patients with early rectal cancer underwent resection of the rectum including lymph nodes and five showed lymph node metastasis (8.6%). Pathology slides of these patients were reviewed by a single pathologist. Demographic and clinical characteristics of these 59 patients were correlated with the existence of nodal metastasis. Formal tests of comparability were carried out by using Fisher's exact test. Logistic regression models were fitted to data to examine possible relationships with 12 covariates measured from each patient and to obtain corresponding odds ratios (as well as a 95% confidence interval for the odds ratios). These covariates included age at surgery, gender, morphology, histology, degree of differentiation, Haggitt's classification for polyps according to the level of invasion, lymphatic and venous invasion, desmoplastic reaction, degree of lymphocytic invasion, presence of lymphoid follicles and presence of infiltrating or pushing margins. A significantly higher rate of lymph node metastasis occurs in the presence of venous invasion (P < 0.01). Venous invasion was present in three of five (60%) patients with lymph node metastasis and only four of 54 (7%) patients without lymph node metastasis. Other variables did not achieve statistical significance. Only the presence of venous invasion was found to be highly significant. The odds ratio of lymph node metastasis increased 18-fold for a patient who had venous invasion compared with a patient who did not. This suggests that the presence of venous invasion in early rectal cancer may provide valuable information to determine which patients would benefit from radical surgery, or adjuvant radiation therapy after sphincter-sparing surgery owing to an increased risk of lymph node metastasis.


Neonicotinoids-from zero to hero in insecticide chemistry.
In recent years, neonicotinoids have been the fastest-growing class of insecticides in modern crop protection, with widespread use against a broad spectrum of sucking and certain chewing pests. As potent agonists, they act selectively on insect nicotinic acetylcholine receptors, their molecular target site. The discovery of neonicotinoids can be considered as a milestone in insecticide research and facilitates greatly the understanding of the functional properties of insect nicotinic acetylcholine receptors. Because of the relatively low risk for non-target organisms and environment, the high target specificity of neonicotinoid insecticides and their versatility in application methods, this important class has to be maintained globally for integrated pest management strategies and insect resistance management programmes. This review comprehensively describes particularly the origin, structure and bonding as well as associated properties of neonicotinoid insecticides.


[A study of left ventricle filling using pulsed Doppler in patients with a high prevalence of atherosclerosis].
We studied the left ventricular diastolic function with Doppler Echocardiography in 61 patients: the first group (A) was conformed by 20 patients with cerebral stroke in evolution (24 to 48 hours) with age between (45 to 63 year old) every one had carotid obstructive lesion shown by angiography. The (B) group had 21 patients with ischemic heart disease and all of them had selective coronary arteriography with atheromatous obstructive lesions (45 to 63 years old); 17 of this patients had previous myocardial infarction. The third group (C) was formed by 20 healthy people between 40 and 62 years of age. We found an increase in the late diastolic peak velocity, in the (A/E) ratio in the early diastolic time; in the early diastolic acceleration and in early diastolic desceleration. The difference between groups (A) and (B) with the (C) was evident (P less than 0.001). We conclude that patients of (A) and (B) groups had reduced left ventricular compliance and the (B) group with evidence of ischemic heart disease show more prominent abnormality.


Molecular basis of phosphatidyl-myo-inositol mannoside biosynthesis and regulation in mycobacteria.
Phosphatidyl-myo-inositol mannosides (PIMs) are unique glycolipids found in abundant quantities in the inner and outer membranes of the cell envelope of all Mycobacterium species. They are based on a phosphatidyl-myo-inositol lipid anchor carrying one to six mannose residues and up to four acyl chains. PIMs are considered not only essential structural components of the cell envelope but also the structural basis of the lipoglycans (lipomannan and lipoarabinomannan), all important molecules implicated in host-pathogen interactions in the course of tuberculosis and leprosy. Although the chemical structure of PIMs is now well established, knowledge of the enzymes and sequential events leading to their biosynthesis and regulation is still incomplete. Recent advances in the identification of key proteins involved in PIM biogenesis and the determination of the three-dimensional structures of the essential phosphatidyl-myo-inositol mannosyltransferase PimA and the lipoprotein LpqW have led to important insights into the molecular basis of this pathway.


Use of aides in occupational therapy practice.
This study examined the use of aides in occupational therapy practice, the supervision and training of aides, and practitioners' attitudes toward the use of aides. A questionnaire was mailed to a systematically selected sample of 510 occupational therapists and occupational therapy assistants. The response rate was 74%. The use of occupational therapy aides is prevalent in a variety of settings. Forty percent of respondents reported that they currently work with aides, whereas 23% reported that they have never worked with aides. Occupational therapy aides perform a variety of tasks and receive various levels and amounts of supervision and training. Of those respondents who delegated specific skilled tasks to aides, 76% reported daily contact for purposes of supervision and training. Nineteen percent who currently work or have worked with aides reported being in a situation where an ethical issue arose related to aide use. On the basis of the data, it is suggested that occupational therapy practitioners would benefit from taking a proactive role in determining how aides can be used to maximize service delivery while maintaining quality services. There also appear to be ethical concerns related to appropriate delegation of tasks to aides and to their supervision and training.


Predictive Value of Serum Infliximab Levels at Induction Phase in Rheumatoid Arthritis Patients.
The Infliximab, has proven effective in treating rheumatoid arthritis (RA). A good clinical response is usually associated with high serum drug levels. Development of antibodies toward Infliximab (ATI) can increase drug clearance, leading to treatment failure. To analyze whether serum Infliximab trough levels (ITL) at the induction phase are associated with Infliximab clearance and clinical outcomes at week(W) 54 and to investigate the association with immunogenicity development. Observational retrospective study in which ITL from 66 RA patients were measured by capture ELISA at W0, W2, W6, W14 and 22. Patients were classified as ITLpos if Infliximab was detectable at W54 and ITLneg otherwise. ATI were assayed by bridging ELISA and by two drug-tolerant assays. ITL cut-off values were established by ROC curves. The association between ITL at early-stage and clearance of Infliximab at W54 was analyzed by univariable and multivariable logistic regression. ITLneg patients (n=25) always had significantly lower Infliximab levels than ITLpos (n=41). An ITL value of 4.4 μg/mL at W6 best predicted W54 Infliximab absence. In the multivariable analysis, only ITL below the cut-off at W6 (OR: 86.6; 95%CI: 6.58-1139.99) and non-use of methotrexate (OR: 6.9; 95%CI: 1.04-45.84) remained significantly associated with W54 Infliximab absence. ATI were more frequent in patients with ITL below the cut-off at W6. In RA, ITL at induction phase are inversely associated with Infliximab clearance and clinical outcomes at W54. ATI was the main reason for low early ITL. A predictive value of ITL at W6 was found as a useful prognostic measure of treatment efficacy.


Machine Learning Improves the Accuracy of Trauma Team Activation Level Assignments in Pediatric Patients.
The assignment of trauma team activation levels can be conceptualized as a classification task. Machine learning models can be used to optimize classification predictions. Our purpose was to demonstrate proof-of-concept for a machine learning tool for predicting trauma team activation levels in pediatric patients with traumatic injuries. Following IRB approval, we retrospectively collected data from the institutional trauma registry and electronic medical record at our Pediatric Trauma Center for all patients (age <18 y) who triggered a trauma team activation (1/2014-12/2021), including: demographics, mechanisms of injury, comorbidities, pre-hospital interventions, numeric variables, and the six "Need for Trauma Intervention (NFTI)" criteria. Three machine learning models (Logistic Regression, Random Forest, Support Vector Machine) were tested 1000 times in separate trials using the union of the Cribari and NFTI metrics as ground-truth (Injury Severity Score >15 or positive for any of 6 NFTI criteria = full activation). Model performance was quantified and compared to emergency department (ED) staff. ED staff had 75% accuracy, an area under the curve (AUC) of 0.73 ± 0.04, and an F1 score of 0.49. The best performing of all machine learning models, the support vector machine, had 80% accuracy, AUC 0.81 ± 4.1e-5, F1 Score 0.80, with less variance compared to other models and ED staff. All machine learning models outperformed ED staff in all performance metrics. These results suggest that data-driven methods can optimize trauma team activations in the ED, with potential improvements in both patient safety and hospital resource utilization. Economic/Decision Analysis or Modeling Studies. II.


Yttrium orthoaluminate nanoperovskite doped with Tm3+ ions as upconversion optical temperature sensor in the near-infrared region.
The thermal sensing capability of the Tm3+-doped yttrium orthoaluminate nanoperovskite in the infrared range, synthetized by a sol-gel method, was studied. The temperature dependence of the infrared upconverted emission bands located at around 705 nm (3F2,3→3H6) and 800 nm (3H4→3H6) of YAP: Tm3+ nanoperovskite under excitation at 1210 nm was analyzed from RT up to 425 K. Calibration of the optical sensor has been made using the fluorescence intensity ratio technique, showing a high sensitivity in the near-infrared compared to other trivalent rare-earth based optical sensors working in the same range. In addition, a second calibration procedure of the YAP: Tm3+ optical sensor was performed by using the FIR technique on the emission band associated to the 3H4→3H6 transition in the physiological temperature range (293-333 K), showing a very high relative sensitivity compared with other rare-earth based optical temperature sensors working in the physiological range. Moreover, the main advantage compared with other optical sensors is that the excitation source and the upconverted emissions do not overlap, since they lie in different biological windows, thus allowing its potential use as an optical temperature probe in the near-infrared range for biological applications.


Navigating the Chemical Space of Multitarget-Directed Ligands: From Hybrids to Fragments in Alzheimer's Disease.
Multitarget drug discovery is one of the hottest topics and most active fields in the search for new molecules against Alzheimer's disease (AD). Over the last 20 years, many promising multitarget-directed ligands (MTDLs) have been identified and developed at a pre-clinical level. However, how to design them in a rational way remains the most fundamental challenge of medicinal chemists. This is related to the foundational question of achieving an optimized activity towards multiple targets of interest, while preserving drug-like properties. In this respect, large hybrid molecules and small fragments are poles apart. In this review article, our aim is to appraise what we have accomplished in the development of both hybrid- and fragment-like molecules directed to diverse AD targets (i.e., acetylcholinesterase, NMDA receptors, metal chelation, BACE-1 and GSK-3β). In addition, we attempt to highlight what are the persistent needs that deserve to be improved and cared for, with the ultimate goal of moving an MTDL to AD clinical studies.


The roles of polymerases ν and θ in replicative bypass of O6- and N2-alkyl-2'-deoxyguanosine lesions in human cells.
Exogenous and endogenous chemicals can react with DNA to produce DNA lesions that may block DNA replication. Not much is known about the roles of polymerase (Pol) ν and Pol θ in translesion synthesis (TLS) in cells. Here we examined the functions of these two polymerases in bypassing major-groove O6-alkyl-2'-deoxyguanosine (O6-alkyl-dG) and minor-groove N2-alkyl-dG lesions in human cells, where the alkyl groups are ethyl, n-butyl (nBu), and, for O6-alkyl-dG, pyridyloxobutyl. We found that Pol ν and Pol θ promote TLS across major-groove O6-alkyl-dG lesions. O6-alkyl-dG lesions mainly induced G→A mutations that were modulated by the two TLS polymerases and the structures of the alkyl groups. Simultaneous ablation of Pol ν and Pol θ resulted in diminished mutation frequencies for all three O6-alkyl-dG lesions. Depletion of Pol ν alone reduced mutations only for O6-nBu-dG, and sole loss of Pol θ attenuated the mutation rates for O6-nBu-dG and O6-pyridyloxobutyl-dG. Replication across the two N2-alkyl-dG lesions was error-free, and Pol ν and Pol θ were dispensable for their replicative bypass. Together, our results provide critical knowledge about the involvement of Pol ν and Pol θ in bypassing alkylated guanine lesions in human cells.


Structure and cohesive energy of dipolar helices.
This paper deals with the investigation of cohesive energy in dipolar helices made up of hard spheres. Such tubular helical structures are ubiquitous objects in biological systems. We observe a complex dependence of cohesive energy on surface packing fraction and dipole moment distribution. As far as single helices are concerned, the lowest cohesive energy is achieved at the highest surface packing fraction. Besides, a striking non-monotonic behavior is reported for the cohesive energy as a function of the surface packing fraction. For multiple helices, we discover a new phase, exhibiting markedly higher cohesive energy. This phase is referred to as ZZ tube consisting of stacked crown rings (reminiscent of a pile of zig-zag rings), resulting in a local triangular arrangement with densely packed filaments parallel to the tube axis.


Novel Design Approaches in the Fabrication of Polymeric Microarray Patches via Micromoulding.
The focus on novel systems for transdermal delivery of therapeutic agents has increased considerably over recent years, as this administration route comes with many advantages. Polymeric microarray patches (MAPs) are minimally invasive devices that enable systemic delivery of a wide range of drugs by overcoming the outer skin barrier. Conventionally, MAPs fabricated by micromoulding have a low needle density. In this study, the performance of hydrogel-forming MAPs cast using novel industrially manufactured micromoulds with a high needle density (600 needles/0.75 cm2) was compared to that of MAPs obtained using conventional moulds with a lower density (196 needles/0.89 cm2). Surrounding holders for micromoulds were designed for time-efficient fabrication of MAPs. The influence of needle densities on mechanical strength, insertion efficiency and in vitro permeation of ibuprofen sodium (IBU) was analysed. Insertion of both MAPs into an artificial skin model and neonatal porcine skin was comparable. No significant difference was observed in permeation studies of IBU (p > 0.05), with a delivery of 8.7 ± 1.7 mg for low-density and 9.5 ± 0.1 mg for high-density MAPs within 24 h. This highlights the potential of these novel micromoulds for manufacturing polymeric MAPs with a higher needle density for future applications.


Short communication: previous semen donors and their views regarding the sharing of information with offspring.
The UK government has decided to introduce, from 2005, rules that will allow donor-conceived persons to have access to identifying information concerning their donor. This has led to many concerns regarding future gamete donor recruitment. Semen donors who had been recruited between 1988 and 2002 were invited to take part in a telephone interview. The interview sought these previous donors' views on issues associated with recruitment, attitudes regarding information sharing and views concerning the offspring. Responses regarding information sharing were compared with their views recorded at the time of recruitment. All 32 donors were recruited altruistically. Eighteen (56%) held the same views concerning the provision of identifying information as they did at the time of recruitment. Of those who had changed their views, eight (25%) expressed a willingness to be more open and four (12%) now wished to be anonymous having previously been unsure. Half of the donors would still have donated if they had been required to be identified to offspring, one-quarter would not have and one-quarter were undecided, although the majority of these said they may have donated under an open system. The study shows that it is possible to recruit identifiable donors at this clinic and this suggests that it may be possible for other clinics to do likewise.


GlyGly-CTERM and rhombosortase: a C-terminal protein processing signal in a many-to-one pairing with a rhomboid family intramembrane serine protease.
The rhomboid family of serine proteases occurs in all domains of life. Its members contain at least six hydrophobic membrane-spanning helices, with an active site serine located deep within the hydrophobic interior of the plasma membrane. The model member GlpG from Escherichia coli is heavily studied through engineered mutant forms, varied model substrates, and multiple X-ray crystal studies, yet its relationship to endogenous substrates is not well understood. Here we describe an apparent membrane anchoring C-terminal homology domain that appears in numerous genera including Shewanella, Vibrio, Acinetobacter, and Ralstonia, but excluding Escherichia and Haemophilus. Individual genomes encode up to thirteen members, usually homologous to each other only in this C-terminal region. The domain's tripartite architecture consists of motif, transmembrane helix, and cluster of basic residues at the protein C-terminus, as also seen with the LPXTG recognition sequence for sortase A and the PEP-CTERM recognition sequence for exosortase. Partial Phylogenetic Profiling identifies a distinctive rhomboid-like protease subfamily almost perfectly co-distributed with this recognition sequence. This protease subfamily and its putative target domain are hereby renamed rhombosortase and GlyGly-CTERM, respectively. The protease and target are encoded by consecutive genes in most genomes with just a single target, but far apart otherwise. The signature motif of the Rhombo-CTERM domain, often SGGS, only partially resembles known cleavage sites of rhomboid protease family model substrates. Some protein families that have several members with C-terminal GlyGly-CTERM domains also have additional members with LPXTG or PEP-CTERM domains instead, suggesting there may be common themes to the post-translational processing of these proteins by three different membrane protein superfamilies.


Clinical impact of minimal residual disease and genetic subtypes on the prognosis of childhood acute lymphoblastic leukemia.
This study analyzed data from two consecutive protocols for children newly diagnosed with acute lymphoblastic leukemia (ALL) to determine the clinical impact of minimal/measurable residual disease (MRD) and recently identified tumor genetic subtypes. Genetic subtypes were determined by sequential approaches including DNA indexing, reverse transcriptase-polymerase chain reaction, multiplex ligation-dependent probe amplification, and RNA-sequencing. MRD was assessed by flow cytometry. The Taiwan Pediatric Oncology Group TPOG-ALL-2013 study enrolled patients who received MRD-directed therapy. The 5-year event-free survival (EFS) and overall survival rates in the 2013 cohort were 77.8% and 86.9% compared to those of the 2002 cohort, which were 62.4% and 76.5%. Among patients treated with MRD-guided therapy, those with ETV6-RUNX1 fusion and high hyperdiploidy had the highest 5-year EFS (91.4% and 89.6%, respectively). The addition of dasatinib improved outcomes in patients with BCR-ABL1 ALL. Recently identified subtypes like DUX4-rearranged, ZNF384-rearranged, MEF2D-rearranged, and PAX5alt subtypes were frequently positive for MRD after remission induction, and these patients consequently received intensified chemotherapy. Treatment intensification according to the MRD improved the outcomes of patients presenting DUX4 rearrangements. In high-risk or very-high-risk subtypes, the TPOG-ALL-2013 regimen did not confer significant improvements compared to TPOG-ALL-2002, and the outcomes of BCR-ABL1-like, MEF2D-rearranged, and KMT2A-rearranged ALL subtypes (in addition to those of T-cell ALL) were not sufficiently good. Novel agents or approaches are needed to improve the outcomes for these patients. The TPOG-ALL-2013 study yielded outcomes superior to those of patients treated in the preceding TPOG-ALL-2002 study. This study provides important data to inform the design of future clinical trials in Taiwan. MRD-directed therapy improved the outcomes for pediatric ALL, especially standard-risk patients. Genomic analyses and MRD might be used together for risk-directed therapy of childhood ALL. Our work provides important data to inform the design of future clinical trials in Taiwan.


Serum high-density lipoprotein cholesterol concentrations in hypo- and hyperthyroidism.
We studied alterations in the concentration of high-density lipoprotein cholesterol and of other lipid categories (total cholesterol, low-density lipoprotein cholesterol, very-low-density lipoprotein cholesterol, and triglycerides) in serum in hypo- and hyperthyroid states, and compared these findings with those of a control group. In the latter group the lipids showed an age-related increase in concentration but with a decrease in the eighth decade in all categories. In untreated hypo- and hyperthyroid subjects, all of the lipid values differed significantly from those of the controls but promptly returned to normal values upon treatment. In hypothyroid patients who are prone to develop coronary heart disease, the concentrations of high-density lipoprotein cholesterol were high, suggesting protection against heart disease, while the ratio to total cholesterol indicated the contrary. The exact opposite of these relationships was seen in hyperthyroid disorders. This apparent paradox suggests that the low-density lipoprotein cholesterol in serum, with its established atherogenic effect, should be given at least equal, if not more, weight than the concentration of high-density lipoprotein cholesterol in serum, with its alleged protective effect.


Processing, physico-chemical, sensory and nutritional evaluation of protein, mineral and vitamin enriched peanut chikki - an Indian traditional sweet.
Chikki, a popular Indian traditional sweet snack prepared from peanut was enriched with protein, minerals and vitamins by incorporating soy protein isolate, calcium carbonate, ferrous fumerate, vitamin A and folic acid to meet the growing demand for health foods. The enriched nutra chikki was evaluated for physico-chemical characteristics such as moisture, texture, peroxide value (PV), fatty acid composition in comparison with control peanut chikki. Nutra chikki, which had initial moisture content of 2-3%, did not alter much up to 60 days of storage at 27 °C and 37 °C. PV increased gradually at 27 °C, whereas at 37 °C, it was higher. Hardness of nutra chikki did not change significantly when stored at 27 °C up to 105 days, but at 37 °C a gradual increase in hardness was observed after 45 days. These results of nutra chikki were similar and comparable with those of control chikki. Nutra chikki had 18% protein, 20% fat, 6.42% Ca, 1.7% Fe, 4000 μg vitamin A and 2660 μg folic acid. Protein digestibility corrected amino acid score of nutra chikki was 0.78 whereas that of control chikki was 0.73. The fatty acid composition of control and nutra chikki was same as both contain peanuts as oil source. The formulation and process parameters for preparation of protein, mineral and vitamin enriched peanut chikki, were standardized. The storage stability and quality parameters of enriched chikki were comparable with those of control chikki.


Protective effect of IAXO-102 on renal ischemia-reperfusion injury in rats.
Ischemia/reperfusion injury (IRI) is a common cause of kidney damage, characterized by oxidative stress and inflammation. In this study, we investigated the potential protective effects of IAXO-102, a chemical compound, on experimentally induced IRI in male rats. The bilateral renal IRI model was used, with 24 adult male rats randomly divided into four groups (N=6): sham group (laparotomy without IRI induction), control group (laparotomy plus bilateral IRI for 30 minutes followed by 2 hours of reperfusion), vehicle group (same as control but pre-injected with the vehicle), and treatment group (similar to control but pre-injected with IAXO-102). We measured several biomarkers involved in IRI pathophysiology using enzyme-linked immunosorbent assay (ELISA), including High mobility group box1 (HMGB1), nuclear factor kappa b-p65 (NF-κB p65), interleukin beta-1 (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), 8-isoprostane, Bcl-2 associated X protein (BAX), heat shock protein 27 (HSP27), and Bcl-2. Statistical analysis was performed using one-way ANOVA and Tukey post hoc tests. Our results showed that IAXO-102 significantly improved kidney function, reduced histological alterations, and decreased the inflammatory response (IL-1, IL-6, and TNF) caused by IRI. IAXO-102 also decreased apoptosis by reducing pro-apoptotic Bax and increasing anti-apoptotic Bcl-2 without impacting HSP27. In conclusion, our findings suggest that IAXO-102 had a significant protective effect against IRI damage in the kidneys.


[Expression of hypoxia-inducible factor-1alpha and matrix metalloproteinase-2 in degenerative lumbar intervertebral disc].
To investigate the expression of hypoxia-inducible factor-1alpha (HIF-1alpha) and matrix metalloproteinase-2 (MMP-2) in degenerative human intervertebral disc. Specimens of the nucleus pulposus of the degenerative discs were obtained from patients with lumbar disc herniation (experimental group), and those from young patients suffering thoracolumbal vertebral bursting fracture served as the control. The experimental group was divided into three subgroups according to the magnetic resonance imaging (MRI) results. The tissue specimens from all the groups were examined for HIF-1alpha and MMP-2 expressions using immunohistochemistry. The tissues in the experimental group showed significantly higher expressions of HIF-1alpha and MMP-2 than those in the control group (P<0.05), and the expressions increased significantly with the severity of the degenerative changes of the intervertebral discs (P<0.05). A positive correlation was found between the expressions of the HIF-1alpha and MMP-2 in degenerative intervertebral discs. HIF-1alpha and MMP-2 may participate in the degeneration of human intervertebral discs, in which process their expressions show a common pattern of enhancement with the progression of the condition.


Effectiveness of over-the-counter nicotine replacement therapy: a qualitative review of nonrandomized trials.
Randomized trials conducted in over-the-counter (OTC) settings have shown that nicotine replacement therapy (NRT) is effective. This paper reviews nonrandomized tests of the effectiveness of OTC NRT. Literature search via computer and other methods located (a) retrospective cohort studies of users versus nonusers of OTC NRT and (b) studies of quit rates before versus after NRT went OTC or before versus after NRT was given free to quitline callers. The methods were too heterogeneous to allow meta-analysis. The results were similar for cohort and pre- versus post-studies. Most of the studies found numerically greater quitting among NRT users than nonusers. Often when NRT was not found effective, other assumed effective treatments (e.g., phone counseling) were also not found effective, suggesting biased or insensitive study methods. Only about half of the studies found statistically greater quitting among NRT users, and the most rigorous studies did not find greater quitting among users. Many studies found selection bias, for example, NRT users are more dependent smokers. Some lines of evidence appear to confirm the effectiveness of OTC NRT, but others do not. We believe further secondary analyses using nonrandomized comparisons are unlikely to resolve this issue due to sensitivity, specificity, and selection bias problems.


Potential Use of Modulators of Oxidative Stress as Add-on Therapy in Patients with Anxiety Disorders.
It is known that an increased oxidative stress is present in a wide range of diseases and, given the vulnerability of the central nervous system, its involvement has been in particular investigated in neurological and psychiatric diseases, including anxiety disorders. In this review, we analyse the studies that have been conducted on the effects of oxidative stress modulators in anxiety, focusing on their possible clinical use. While preclinical studies have shown a clear anxiolytic-like effect of different oxidative stress modulators, less significant results have been obtained from clinical studies. After having reviewed the possible reasons for the discrepancy between preclinical and clinical data, we encourage further studies aimed at better investigating the utility of the modulation of oxidative stress in humans, as an adjunctive therapy of the traditional integrated psychotherapeutic and pharmacological approach.


Application of IS900 PCR for detection of Mycobacterium avium subsp. paratuberculosis directly from raw milk.
A polymerase chain reaction (PCR)-based assay was developed for detection of insertion sequence 900 (IS900) of Mycobacterium avium subsp. paratuberculosis in raw milk. This IS900 PCR assay included DNA extraction and PCR assay using commercially available kits. The DNA extraction and PCR assay were optimized to detect the IS900 sequence directly from raw milk. The IS900 PCR assay was evaluated by inoculating raw bulk milk and Middlebrook's 7H9 broth with 0 to 10(8) cfu/ml of each of four American Type Culture Collection strains of M. paratuberculosis. Under experimental conditions, both milk culture on Herrold's egg yolk medium slants, and IS900 PCR could detect 10 to 100 cfu/ml of M. paratuberculosis. Detection of M. paratuberculosis by IS900 PCR was consistent (24/24 PCR assays) when about 100 cfu/ml were present, whereas detection was variable (12/24 PCR assays) at concentrations as low as 10 cfu/ml. Based on the findings of the experimental study, IS900 PCR was further evaluated with pooled quarter milk samples from 211 cows from five herds with known history of Johne's disease. Out of 211 animals examined, nine (4%) and 69 (33%) were positive for M. paratuberculosis by milk culture and IS900 PCR from milk, respectively. A total of 20 bulk tank milk sample aliquots (one sample, four aliquots from each herd) were also examined, of which 10 (50%) were positive for M. paratuberculosis by IS900 PCR. By contrast, only one out of 20 (5%) bulk tank milk sample aliquots was positive by culture. The IS900 PCR amplified product of 229-bp obtained on testing of quarter milk and bulk tank milk samples was confirmed to be the IS900 of M. paratuberculosis by DNA sequence analysis. The results of this study suggest that M. paratuberculosis can be detected directly from quarter milk and bulk tank milk by IS900 PCR.


Liquid-crystalline elastomer-nanoparticle hybrids with reversible switch of magnetic memory.
A stimuli-responsive material is synthesized that combines the actuation potential of liquid-crystalline elastomers with the anisotropic magnetic properties of ellipsoidal iron oxide nanoparticles. The resulting nanocomposite exhibits unique shape-memory features with magnetic information, which can be reversibly stored and erased via parameters typical of soft materials, such as high deformations, low stresses, and liquid-crystalline smectic-isotropic transition temperatures.