The effect of intravaginal applied GnRH-agonist on the time of ovulation and subsequent reproductive performance of weaned multiparous sows.
In order to prove the effect of 'fixed time insemination' and insemination at standing oestrus after post-weaning application of GnRH, in a Croatian large breeding unit, 502 sows were assigned to three groups and were artificially inseminated (AI) at their first post-weaning oestrus as many times as they stand, in 24-h intervals. The groups were treated as follows: group 1 (control, n = 160) were AI during their standing reflex; group 2 ['GnRH-fixed time insemination' (GnRH-FT-AI), n = 175] were AI, independent of detection of oestrus and following administration of GnRH-agonist at 96 h post-weaning; group 3 [GnRH insemination at standing oestrus (GnRH-OE-AI), n = 167] the animals were GnRH-agonist treated as group 2 and were AI at their standing reflex. Pre-trial daily average lactational feed intake, average daily feed intake from weaning to oestrus, oestrus within 6 days post-weaning (%), ovulation within 6 days post-weaning (%), weaning-to-oestrus interval (h), duration of oestrus (h), follicle size (mm), interval from oestrus to ovulation (h), subsequent day 24 pregnancy rate (%), farrowing rate (%) and total pigs born were evaluated. Pre-trial average daily lactational voluntary feed intake was 7.1 +/- 0.08 kg in group 1, 7.0 +/- 0.07 kg in group 2 and 7.1 +/- 0.17 kg in group 3 (p > 0.05). Average voluntary daily feed intake from weaning to oestrus was 5.1 +/- 0.3 kg in group 1, 5.2 +/- 0.5 kg in group 2 and 5.2 +/- 0.19 kg in group 3 (p > 0.05). Oestrus was detected within 6 days post-weaning in 134 (83.8%) in control, 164 (93.7%) in GnRH-FT-AI and 155 (92.8%) animals in GnRH-OE-AI groups (p = 0.05). Follicle size did not differ (p > 0.05) among the groups. In control 82.8%, in GnRH-FT-AI 91.5% and in GnRH-OE-AI 91.0% of the sows ovulated within 6 days post-weaning (p = 0.04), and had 80.6, 90.9 and 89.7% 24-day pregnancy rates (p = 0.16), respectively. In GnRH-FT-AI group 90.2%, in GnRH-OE-AI sows 89.7%, in control animals 79.9% farrowing rates were recorded (p = 0.17). Weaning to oestrus interval was 113.1 h in control, 114.1 h in GnRH-FT-AI and 112.6 h GnRH-OE-AI (p > 0.05). Duration of oestrus was significantly shorter in GnRH-FT-AI (44.9 h) and GnRH-OE-AI (48.1 h) animals, compared with the control (62.9 h) sows (p = 0.001). Similarly, the interval from oestrus to ovulation revealed significant (p = 0.004) differences between the groups (control 44.1 h, GnRH-OE-AI 34.1 h and GnRH-FT-AI 32.9 h). GnRH-FT-AI (12.5) and GnRH-OE-AI (12.6) sows had significantly higher (p = 0.01) number of total pigs born (n = 10.4) compared with control sows. GnRH-agonist-gel treatment to the sow shortens duration of oestrus, the interval from oestrus to ovulation, and may eliminate the need for oestrus detection in the hands of skilled personnel.


Rabbit classical eyeblink conditioning is altered by brief cerebellar cortical stimulation.
A pair of studies examined how cortical intracerebellar stimulation (ICS) affects eyeblink conditioning in the rabbit. Rabbits were implanted with chronic bipolar stimulating electrodes in the cell body layers of cerebellar lobule H-VI. Brief (40 ms) trains of intracranial stimulation (100 Hz, 250 microA) were delivered during training trials [forward pairings of a tone-conditioned stimulus (CS) with an air puff unconditioned stimulus (US)]. In Experiment 1, the onset of ICS varied randomly within sessions. US-onset-coincident ICS proved detrimental to the maintenance of conditioning [measured as the percentage of trials on which conditioned responses (CRs) were made] compared to ICS that ended 60 ms before US onset. Based on these findings, a second experiment compared a group trained with ICS consistently delivered at US onset to groups trained with ICS consistently delivered either at CS onset or between the two stimuli, as well as to unstimulated control subjects. Animals receiving CS- or US-coincident ICS learned slowest, whereas animals receiving middle stimulation learned more quickly than all other groups. In both Experiments 1 and 2, highly trained animals produced blinks in direct response to the stimulation. These data are discussed in terms of a new hypothesis concerning interactions between cerebellar cortex and the deep cerebellar nuclei during eyeblink conditioning--a rebound from inhibition hypothesis.


Comparison of Self-Etch Primers with Conventional Acid Etching System on Orthodontic Brackets.
The self-etching primer system consists of etchant and primer dispersed in a single unit. The etching and priming are merged as a single step leading to fewer stages in bonding procedure and reduction in the number of steps that also reduces the chance of introduction of error, resulting in saving time for the clinician. It also results in smaller extent of enamel decalcification. To compare the Shear Bond Strength (SBS) of orthodontic bracket bonded with Self-Etch Primers (SEP) and conventional acid etching system and to study the surface appearance of teeth after debonding; etching with conventional acid etch and self-etch priming, using stereomicroscope. Five Groups (n=20) were created randomly from a total of 100 extracted premolars. In a control Group A, etching of enamel was done with 37% phosphoric acid and bonding of stainless steel brackets with Transbond XT (3M Unitek, Monrovia, California). Enamel conditioning in left over four Groups was done with self-etching primers and adhesives as follows: Group B-Transbond Plus (3M Unitek), Group C Xeno V+ (Dentsply), Group D-G-Bond (GC), Group E-One-Coat (Coltene). The Adhesive Remnant Index (ARI) score was also evaluated. Additionally, the surface roughness using profilometer were observed. Mean SBS of Group A was 18.26±7.5MPa, Group B was 10.93±4.02MPa, Group C was 6.88±2.91MPa while of Group D was 7.78±4.13MPa and Group E was 10.39±5.22MPa respectively. In conventional group ARI scores shows that over half of the adhesive was remaining on the surface of tooth (score 1 to 3). In self-etching primer groups ARI scores show that there was no or minor amount of adhesive remaining on the surface of tooth (score 4 and 5). SEP produces a lesser surface roughness on the enamel than conventional etching. However, statistical analysis shows significant correlation (p<0.001) of bond strength with surface roughness of enamel. All groups might show clinically useful SBS values and Transbond XT can be successfully used for bracket bonding after enamel conditioning with any of the SEPs tested. The SEPs used in Groups C (Xeno V+) and D (G-Bond) have significantly lowered SBS. Although, the values might still be clinically acceptable.


Race terminology in the field of psychology: Acknowledging the growing multiracial population in the U.S.
The field of psychology is coming toward a critical juncture; scholars are increasingly recognizing that race, ethnicity, and culture play important roles in their fields of study, but do not always have the language to integrate race and culture into their own work. Furthermore, common conceptions of race may systematically exclude those from multiple racial and ethnic backgrounds in favor of fixed and discrete racial categories that ultimately perpetuate white supremacy. Meanwhile, as the Multiracial population of the U.S. is growing at an unprecedented rate, psychologists need language to acknowledge this population in their studies and pursue research to advance the field's knowledge of this diverse group and its many subpopulations. In an attempt to educate psychologists across subfields and disciplines, we provide a detailed account of preferred terms related to race and ethnicity with emphasis on ways to think about and talk about Multiracial populations in the United States. While preferred terms may change across time, the aim of this article is to provide psychologists with the tools to begin nuanced and necessary discussions about how race informs their research and the populations they work with in uniform and nonstigmatizing ways. By highlighting terminology related to those of multiple racial and ethnic backgrounds, we demystify and legitimize these rapidly growing but often hidden populations. Different perspectives on various terms are provided throughout to set psychologists on the path to beginning more race-conscious conversations and scientific inquiries into the experiences of Multiracial Americans and those from other marginalized racial-ethnic groups. (PsycInfo Database Record (c) 2022 APA, all rights reserved).


Nutritional physiology of mahi-mahi (Coryphaena hippurus): Postprandial metabolic response to different diets and metabolic impacts on swim performance.
Migratory pelagic fish species, such as the mahi-mahi (Coryphaena hippurus), must balance numerous metabolic demands simultaneously in order to survive in a challenging oceanic environment. Energetic support for such demands comes from a variety of natural prey items in the wild and can come from manufactured pelletized feed in captivity. This study quantified postprandial metabolism, commonly referred to as specific dynamic action (SDA), over time in adult mahi-mahi (706±25g; 38±0.7cm FL) in response to satiation feeding using three different natural and manufactured diets. Results indicate that during satiation feeding the amount of food ingested is dictated by energy content rather than prey mass, regardless of moisture content of the diet. Ingested meal energy did not differ significantly across groups (473±45kJ), nor did the duration of SDA (36±2.1h). Satiation feeding levels ranged from 2.9-16.2% bodyweight depending on the diet. Peak SDA and SDA magnitude were both significantly decreased in response to dry pelletized diet compared to the natural forage diets, despite equivalent energy consumption. Swim performance and maximum metabolic rate were not impacted significantly in satiation fed fish compared to unfed fish, supporting the evidence that mahi-mahi are able to maintain multiple metabolic demands at one time without compromising performance.


Anesthesia of Tibetan yak (Bos grunniens) using thiafentanil - xylazine and carfentanil - xylazine.
The use of 0.025 +/- 0.012 mg/kg (median +/- interquartile range) thiafentanil with 0.15 +/- 0.03 mg/kg xylazine (TX) and 0.011 +/- 0.0015 mg/kg carfentanil with 0.25 +/- 0.093 mg/kg xylazine (CX), with dosages based on estimated bodyweight, was used in the anesthesia of 37 Tibetan yak (Bos grunniens) housed within a drive-through animal park setting. The median time to lateral recumbency was 5 and 7 min for each group, respectively. With the addition of propofol in 8 CX animals and 17 TX animals, the anesthetic plane was suitable for a wide range of procedures. The median time to standing recovery following administration of naltrexone was 4 +/- 3.5 min with TX and 7 +/- 1.5 min with CX. There was one fatality and one case of renarcotization in the TX group. Overall, the dosages used in the study provided a reliable and useful anesthetic induction protocol, with TX animals demonstrating a more rapid induction and recovery with less cardiac depression than CX animals.


A mutant gene that increases gibberellin production in brassica.
A single gene mutant (elongated internode [ein/ein]) with accelerated shoot elongation was identified from a rapid cycling line of Brassica rapa. Relative to normal plants, mutant plants had slightly accelerated floral development, greater stem dry weights, and particularly, increased internode and inflorescence elongation. The application of the triazole plant growth retardant, paclobutrazol, inhibited shoot elongation, returning ein to a more normal phenotype. Conversely, exogenous gibberellin A(3) (GA(3)) can convert normal genotypes to a phenotype resembling ein. The content of endogenous GA(1) and GA(3) were estimated by gas chromatography-selected ion monitoring using [(2)H]GA(1), as a quantitative internal standard and at day 14 were 1.5- and 12.1-fold higher per stem, respectively, in ein than in normal plants, although GA concentrations were more similar. The endogenous levels of GA(20) and GA(1), and the rate of GA(19) metabolism were simultaneously analyzed at day 7 by feeding [(2)H(2)]GA(19) and measuring metabolites [(2)H(2)]GA(20) and [(2)H(2)]GA(1) and endogenous GA(20) and GA(1), with [(2)H(5)]GA(20) and [(2)H(5)]GA(1) as quantitative internal standards. Levels of GA(1) and GA(20) were 4.6- and 12.9-fold higher, respectively, and conversions to GA(20) and GA(1) were 8.3 and 1.3 times faster in ein than normal plants. Confirming the enhanced rate of GA(1) biosynthesis in ein, the conversion of [(3)H]GA(20) to [(3)H]GA(1) was also faster in ein than in the normal genotype. Thus, the ein allele results in accelerated GA(1) biosynthesis and an elevated content of endogenous GAs, including the dihydroxylated GAs A(1) and A(3). The enhanced GA production probably underlies the accelerated shoot growth and development, and particularly, the increased shoot elongation.


Mechanisms of Antibiotic Tolerance in Mycobacterium avium Complex: Lessons From Related Mycobacteria.
Mycobacterium avium complex (MAC) species are the most commonly isolated nontuberculous mycobacteria to cause pulmonary infections worldwide. The lengthy and complicated therapy required to cure lung disease due to MAC is at least in part due to the phenomenon of antibiotic tolerance. In this review, we will define antibiotic tolerance and contrast it with persistence and antibiotic resistance. We will discuss physiologically relevant stress conditions that induce altered metabolism and antibiotic tolerance in mycobacteria. Next, we will review general molecular mechanisms underlying bacterial antibiotic tolerance, particularly those described for MAC and related mycobacteria, including Mycobacterium tuberculosis, with a focus on genes containing significant sequence homology in MAC. An improved understanding of antibiotic tolerance mechanisms can lay the foundation for novel approaches to target antibiotic-tolerant mycobacteria, with the goal of shortening the duration of curative treatment and improving survival in patients with MAC.


["Unfair" effect of physical activity. Physically inactive individuals must exercise for a longer period to achieve the preventive targets].
The capacity to turnover energy during exercise is regulated by the maximal aerobic capacity. Advices regarding physical activity must therefore be modified and individualised based on the maximal aerobic capacity of the patient. Initially, physical training of patients with low capacity to turn over energy will make it more difficult for the patient to achieve goals of disease prevention and weight reduction. In addition, physical inactive subjects are more negative to physical exercise than physically active subjects. However, physically inactive subjects must be informed that they can increase their maximal aerobic capacity by physical training rapidly. In addition, they must be informed that it takes 10-20 weeks of regular physical endurance training to achieve increased vitality and a more positive attitude towards physical activity, and furthermore, that it is necessary with physical endurance training 2-4 times on a weekly basis to maintain positive effects of physical training.


Effect of aging on hedonic appreciation of pleasant and unpleasant odors.
Does hedonic appreciation evolve differently for pleasant odors and unpleasant odors during normal aging? To answer this question we combined psychophysics and electro-encephalographic recordings in young and old adults. A first study showed that pleasant odorants (but not unpleasant ones) were rated as less pleasant by old adults. A second study validated this decrease in hedonic appreciation for agreeable odors and further showed that smelling these odorants decreased beta event-related synchronization in aged participants. In conclusion, the study offers new insights into the evolution of odor hedonic perception during normal aging, highlighting for the first time a change in processing pleasant odors.


[High-quality hospital discharge summaries - general practitioners expectations].
Hospital discharge summaries ensure treatment continuity after hospital discharge. In Switzerland discharge letters are a celebrated custom and a tool for training young colleagues. The primary purpose is to guarantee high-quality care of patients treated by hospital staff and general practitioners. From the perspective of the patient's general practitioner discharge summaries should convey current and accurate medically important patient data to the physician responsible for follow-up care. In the era of highly developed electronic data transfer and introduction of diagnose related groups (DRGs), it will be necessary to transmit hospital discharge information selectively to different target groups. Nevertheless data protection and medical secret must be complied with.


Acute symptoms associated with antineoplastic drug handling among nurses.
Antineoplastic drug handling in the absence of adequate protective measures has been associated with biological uptake of the drugs among pharmacists and nurses. This study investigated the association between occupational exposure to antineoplastics and the presence of acute symptoms in a nationwide sample of 2,048 nurses and nurses' aides. Reported skin contact with the drugs was associated with a small but statistically significant increase in reported symptoms. Although number of doses handled and extent of protection used were significantly associated with number of symptoms, their effect was not independent of skin contact.


PK assays for antibody-drug conjugates: case study with ado-trastuzumab emtansine.
Antibody-drug conjugates (ADCs) combine the characteristics of large-molecule biologics and small-molecule drugs and are heterogeneous mixtures that can biotransform in vivo, resulting in additional complexity. ADC bioanalytical strategies require novel analytical methods, as well as existing large- and small-molecule methods. Because ADCs in late-stage clinical development are relatively new, regulatory guidelines and standard industry best practices for developing strategies for bioanalytical PK assays are still being established. A PK assay strategy was developed that included comprehensive novel reagent and assay characterization approaches for the ADC ado-trastuzumab emtansine (T-DM1). The bioanalytical strategy was successfully applied to the drug development of T-DM1 and ensured that key analytes were accurately measured in support of nonclinical and clinical development.


Biology and oncogenicity of the Kaposi sarcoma herpesvirus K1 protein.
The Kaposi sarcoma-associated herpesvirus (KSHV), or human herpesvirus 8, is a gammaherpesvirus etiologically linked to the development of Kaposi sarcoma, primary effusion lymphomas, and multicentric Castleman disease in humans. KSHV is unique among other human herpesviruses because of the elevated number of viral products that mimic human cellular proteins, such as a viral cyclin, a viral G protein-coupled receptor, anti-apoptotic proteins (e.g., v-bcl2 and v-FLIP), viral interferon regulatory factors, and CC chemokine viral homologues. Several KSHV products have oncogenic properties, including the transmembrane K1 glycoprotein. KSHV K1 is encoded in the viral ORFK1, which is the most variable portion of the viral genome, commonly used to discriminate among viral genotypes. The extracellular region of K1 has homology with the light chain of lambda immunoglobulin, and its cytoplasmic region contains an immunoreceptor tyrosine-based activation motif (ITAM). KSHV K1 ITAM activates several intracellular signaling pathways, notably PI3K/AKT. Consequently, K1 expression inhibits proapoptotic proteins and increases the life-span of KSHV-infected cells. Another remarkable effect of K1 activity is the production of inflammatory cytokines and proangiogenic factors, such as vascular endothelial growth factor. KSHV K1 immortalizes primary human endothelial cells and transforms rodent fibroblasts in vitro; moreover, K1 induces tumors in vivo in transgenic mice expressing this viral protein. This review aims to consolidate and discuss the current knowledge on this intriguing KSHV protein, focusing on activities of K1 that can contribute to the pathogenesis of KSHV-associated human cancers.


Controlled Site-Directed Assembly of Antibodies by Their Oligosaccharide Moieties onto APTES Derivatized Surfaces.
A convenient and efficient method for the site-directed incorporation of aldehydes generated on the oligosaccharide moieties at the C-terminal of immunoglobulin (IgG) using NaIO4 oxidation reaction is explored as a means of ensuring controlled assembly of IgG antibodies onto aminopropyltriethoxylsilane (APTES) derivatized silicon wafer surfaces. The orientation and antigen binding capacity (AgBC) of site-directly assembled IgG antibodies on derivatized surfaces were investigated using atomic force microscopy (AFM) and enzyme immunoassay (EIA), respectively. A major difference in preferential orientation is observed when the incubation of derivatized surfaces with oxidized IgG molecules is compared in two different kinds of buffer solutions. We obtained the stable and homogeneous IgG layer without loss of the AgBC on the APTES derivatized surface using the controlled incubation condition. Copyright 1999 Academic Press.


[Proportion of schizophrenia in inpatient treatment morbidity of a defined catchment area].
The results of a survey performed on a specific date and an analysis of the discharge files are used as a starting point to discuss the large proportion of schizophrenics among the patients receiving ward treatment at hospitals. Many of these are chronic, long-term patients whose chances of being discharged decrease with increasing duration of hospitalisation. In contrast, instances of hospitalisation of acute schizophrenia cases in the region concerned has decreased considerably during the past 20 years as a result of the more consistent use of psychopharmaceutics and improved dispensary treatment. These modern forms of therapy can only yield optimal results, however, if all those concerned with rehabilitation show a good portion of sociopsychiatric involvement and nonmedical institutions also help with the therapy scheme.


Supercritical CO₂ extraction of oil, fatty acids and flavonolignans from milk thistle seeds: Evaluation of their antioxidant and cytotoxic activities in Caco-2 cells.
The optimal conditions of supercritical carbon dioxide (SC-CO2) (160-220 bars, 40-80 °C) technology combined with co-solvent (ethanol), to recover oil, flavonolignans (silychristin, silydianin and silybinin) and fatty acids from milk thistle seeds, to be used as food additives and/or nutraceuticals, were studied. Moreover, the antioxidant and cytotoxic activities of the SC-CO2 oil seeds extracts were evaluated in Caco-2 carcinoma cells. Pressure and temperature had a significant effect on oil and flavonolignans recovery, although there was not observed a clear trend. SC-CO2 with co-solvent extraction at 220 bars, 40 °C was the optimum treatment to recover oil (30.8%) and flavonolignans from milk thistle seeds. Moreover, linoleic (47.64-66.70%), and oleic (19.68-24.83%) acids were the predominant fatty acids in the oil extracts recovered from milk thistle under SC-CO2. In addition, SC-CO2 extract showed a high antioxidant activity determined by DPPH and ABTS tests. Cytotoxic activities of silychristin, silydianin and silybinin and the obtained SC-CO2 extract (220 bars, 40 °C) were evaluated against Caco-2 cells. The SC-CO2 extract inhibited the proliferation of Caco-2 cells in a dose-responsive manner and induced the highest percentage of mortality of Caco-2 cells (from 43 to 71% for concentrations from 10 up to 100 μg/ml of SC-CO2 oil seeds).


Characterization of natural organic matter (NOM) derived from sewage sludge compost. Part 1: chemical and spectroscopic properties.
In this study changes in the properties of natural organic matter (NOM) were studied during composting of sewage sludge in a laboratory experiment using the pile method. Typical physicochemical parameters were measured during 53 days of composting including humic fractions. The effects of humification on the molecular properties of humic acids (HA) were investigated by 13C CP/MAS NMR spectroscopy. On the basis of chemical analyses, 53 days of composting sewage sludge with structural material can be divided into three phases: (i) domination of rapid decomposition of non-humic, easily biodegradable organic matter (two to three weeks), (ii) domination of organic matter humification and formation of polycondensed, humic-like substances (the next two weeks), (iii) stabilization of transformed organic material and weak microbial activity. Spectroscopic characterization (13C NMR) of compost humic acids reveals changes in their structures during maturation. The changes are highly correlated with the processes taking place in bulk compost.


Effect of an on-sight lead on the physiological and psychological responses to rock climbing.
Rock climbing is a multi-discipline activity that encompasses forms such as bouldering, top roping and lead climbing on natural and artificial climbing surfaces. A major focus of research has been explanation of physiological functioning. More recent research indicates that anxiety levels are elevated for less experienced climbers and in response to lead climbing ascents. Research regarding the demands of rock climbing has placed a lesser focus on the interaction of psychological and physiological factors. The objective of this study was to examine the effects of an on-sight lead climb on the physiological and psychological demands of the climb in comparison with a subsequent lead climb. Ten intermediate level climbers volunteered to complete the two climbing trials, on-sight lead climb (OSLC) and second lead climb (LC2). Climb time, lactate concentrations (baseline, pre climb, post climb and 15 min post climb), heart rate (1 min pre climb, peak HR, 1 min post climb and average climb across the duration of the climb), oxygen consumption, pre climb anxiety (CSAI-2R) were assessed for each climber for both trials. Results indicated that there were significant differences in self reported pre climb somatic and cognitive anxiety (t(9) = 2.79, p = 0.01, t(9) = 1.94, p = 0.043), climb time (t(9) = 3.07, p = 0.0052) and post climb lactate concentrations between the climbs (t(9) = 2.58, p = 0.015). These results indicate that psychological as well as physiological stress impact upon the response to rock climbing. The higher anxiety levels associated with an OSLC are likely to have influenced the physiological responses for the intermediate climbers in this study. Future studies should take into account the type of climbing, experience of climbers and the number of ascents as well as taking into account the interaction between physiological and psychological factors in response to rock climbing. Key pointsFor intermediate climbers, there are significant differences in physiological and psychological responses to on-sight lead and subsequent lead climb.There was an increased psychological and physiological load for leading a climb for the first time (on-sight).These results indicate that the type of climbing should be taken into account when interpreting results from climbing studies with intermediate or recreational climbers.


Thermogenic responsiveness to beta-adrenergic stimulation is augmented in exercising versus sedentary adults: role of oxidative stress.
Beta-adrenergic receptor (beta-AR) modulation of resting and postprandial energy expenditure (EE) is augmented in regularly exercising compared with sedentary adults, but the underlying physiological mechanisms are unknown. Differences in thermogenic responsiveness to beta-AR stimulation, perhaps secondary to reactive oxygen species (ROS) bioactivity, may be involved. To determine habitual exercise-related differences in beta-AR thermogenic responsiveness and the possible influence of ROS, we measured the percentage increase in EE (DeltaEE%; indirect calorimetry, ventilated hood method) above resting EE in response to non-specific beta-AR stimulation (intravenous isoproterenol (isoprenaline): 6, 12 and 24 ng (kg fat-free mass)-1 min-1) in 25 sedentary (11 males; 51+/-4 years; body mass index 25.0+/-0.8 kg m-2, maximal oxygen uptake 29+/-1 ml kg-1 min-1 (mean+/-s.e.m.)) and 14 habitually aerobic exercising (9 males, 46+/-6 years, 23.1+/-0.7 kg m-2, 44+/-3 ml kg-1 min-1) healthy adults under normal (control) conditions and during acute intravenous administration of a potent antioxidant, ascorbic acid (vitamin C; 0.04 g (kg fat-free mass)-1). DeltaEE% was greater (P=0.02) in the habitually exercising (8.6+/-1.2, 12.9+/-1.2, 20.0+/-1.4) versus sedentary (6.3+/-0.7, 10.4+/-0.8, 16.0+/-1.0) adults. Ascorbic acid increased (P=0.01) DeltaEE% only in the sedentary adults (to 9.5+/-0.9, 12.4+/-0.7, 18.5+/-0.8), abolishing baseline group differences. DeltaEE% was not related to the amount of body fat, sex, or any other baseline characteristic. Thermogenic responsiveness to beta-AR stimulation is augmented in habitually exercising adults. The mechanism is ascorbic acid dependent, suggesting that it may be linked to decreased ROS bioactivity. Our findings advance a novel mechanism by which habitual physical activity may modulate EE in humans, with potential implications for energy balance and body weight control.


Electron beam inactivation of selected microbial pathogens and indicator organisms in aerobically and anaerobically digested sewage sludge.
Microbial pathogens in municipal sewage sludges need to be inactivated prior to environmental disposal. The efficacy of high energy (10 MeV) e-beam irradiation to inactivate a variety of selected microbial pathogens and indicator organisms in aerobically and anaerobically digested sewage sludge was evaluated. Both bacterial and viral pathogens and indicator organisms are susceptible to e-beam irradiation. However, as expected there was a significant difference in their respective e-beam irradiation sensitivity. Somatic coliphages, bacterial endospores and enteric viruses were more resistant compared to bacterial pathogens. The current US EPA mandated 10 kGy minimum dose was capable of achieving significant reduction of both bacterial and viral pathogens. Somatic coliphages can be used as a microbial indicator for monitoring e-beam processes in terms of pathogen inactivation in sewage sludges.


Analysis of a single α-synuclein fibrillation by the interaction with a protein nanopore.
The formation of an α-synuclein fibril is critical in the pathogenesis of Parkinson's disease. The native unfolded α-synuclein monomer will translocate through an α-hemolysin nanopore by applied potential at physiological conditions in vitro. Applying a potential transformed α-synuclein into a partially folded intermediate, which was monitored by capture inside the vestibule of an α-hemolysin nanopore with a capture current of 20 ± 1.0 pA. The procedure involves the critical early stage of α-synuclein structural transformation. Further elongation of the intermediate produces a block current to 5 ± 0.5 pA. It is revealed that the early stage fibril of α-synuclein inside the nanopore is affected by intrapeptide electrostatic interaction. In addition, trehalose cleared the fibrillation by changing the surface hydrophobic interaction of A53T α-synuclein, which did not show any inhibition effect from WT α-synuclein. The results proved that the interpeptide hydrophobic interactions in the elongation of A53T α-synuclein protofilaments can be greatly weakened by trehalose. This suggests that trehalose inhibits the interpeptide interaction involved in protein secondary structure. The hydrophobic and electrostatic interactions are associated with an increase in α-synuclein fibrillation propensity. This work provides unique insights into the earliest steps of the α-synuclein aggregation pathway and provides the potential basis for the development of drugs that can prevent α-synuclein aggregation at the initial stage.


Source of tunable radiation based on Ti(3+):Al(2)O(3) crystals pumped by a copper-vapor laser.
A source of tunable coherent radiation based on the use of Ti(3+):Al(2)O(3) crystals pumped by Cu-vapor laser radiation is studied experimentally. This source is promising for applications to atmospheric sensing as well as for use in applied optics and spectroscopy studies. Energy, kinetic, spectral, and spatial characteristics of lasing in nonselective and selective cavities and in the master oscillator power amplifier system are measured. Based on the latter scheme an experimental model of the source is developed. It permits continuously tunable radiation to be obtained in two spectral regions, i.e., within the fundamental band from 680 to 980 nm with a maximum average output power of ~2.6 W and in its second harmonic range from 340 to 490 nm with a maximum average output power of 450 mW. The linewidth of the emission is ~2 pm everywhere in the operating wavelength ranges, and the beam divergence is close to that of the diffraction. The pulse repetition rate is 5 kHz. A further increase in the output power is possible.


Morusin Suppresses Cancer Cell Invasion and MMP-2 Expression through ERK Signaling in Human Nasopharyngeal Carcinoma.
The most important cause of treatment failure of nasopharyngeal carcinoma (NPC) patients is metastasis, including regional lymph nodes or distant metastasis, resulting in a poor prognosis and challenges for treatment. In the present study, we investigated the in vitro anti- tumoral properties of morusin on human nasopharyngeal carcinoma HONE-1, NPC-39, and NPC-BM cells. Our study revealed that morusin suppressed the migration and invasion abilities of the three NPC cells. Gelatin zymography assay and Western blotting demonstrated that the enzyme activity and the level of matrix metalloproteinases-2 (MMP-2) protein were downregulated by the treatment of morusin. Mitogen-activated protein kinase proteins were examined to identify the signaling pathway, which showed that phosphorylation of ERK1/2 was inhibited after the treatment of morusin. In summary, our data showed that morusin inhibited the migration and invasion of NPC cells by suppressing the expression of MMP-2 by downregulating the ERK1/2 signaling pathway, suggesting that morusin may be a potential candidate for chemoprevention or adjuvant therapy of NPC.


Proteolytic cleavage and truncation of NDRG1 in human prostate cancer cells, but not normal prostate epithelial cells.
NDRG1 (N-myc downstream regulated gene-1) is a metastasis suppressor that is down-regulated in prostate cancer. NDRG1 phosphorylation is associated with inhibition of metastasis and Western blots indicate two bands at ~41 and ~46 kDa. Previous investigations by others suggest the higher band is due to NDRG1 phosphorylation. However, the current study using a dephosphorylation assay and the Phos-tag (phosphate-binding tag) SDS/PAGE assay, demonstrated that the 46 kDa NDRG1 protein band was not due to phosphorylation. Further experiments showed that the NDRG1 protein bands were not affected upon glycosidase treatment, despite marked effects of these enzymes on the glycosylated protein, fetuin. Analysis using RT-PCR (reverse transcriptase-PCR) demonstrated only a single amplicon, and thus, the two bands could not result from an alternatively spliced NDRG1 transcript. Western-blot analysis of prostate cancer cell lysates identified the 41 kDa band to be a truncated form of NDRG1, with MS confirming the full and truncated proteins to be NDRG1. Significantly, this truncated protein was not present in normal human PrECs (prostate epithelial cells). Western-blot analysis using anti-NDRG1 raised to its N-terminal sequence failed to detect the truncated protein, suggesting that it lacked N-terminus amino acids (residues 1-49). Sequence analysis predicted a pseudotrypsin protease cleavage site between Cys49-Gly50. Such cleavage of NDRG1 in cancer cells may result in loss of NDRG1 tumour suppressive activity.


Genomic comparison of plant pathogenic and nonpathogenic Serratia marcescens strains by suppressive subtractive hybridization.
Cucurbit yellow vine disease (CYVD) is caused by disease-associated Serratia marcescens strains that have phenotypes significantly different from those of nonphytopathogenic strains. To identify the genetic differences responsible for pathogenicity-related phenotypes, we used a suppressive subtractive hybridization (SSH) strategy. S. marcescens strain Z01-A, isolated from CYVD-affected zucchini, was used as the tester, whereas rice endophytic S. marcescens strain R02-A (IRBG 502) was used as the driver. SSH revealed 48 sequences, ranging from 200 to 700 bp, that were present in Z01-A but absent in R02-A. Sequence analysis showed that a large proportion of these sequences resembled genes involved in synthesis of surface structures. By construction of a fosmid library, followed by colony hybridization, selection, and DNA sequencing, a phage gene cluster and a genome island containing a fimbrial-gene cluster were identified. Arrayed dot hybridization showed that the conservation of subtracted sequences among CYVD pathogenic and nonpathogenic S. marcescens strains varied. Thirty-four sequences were present only in pathogenic strains. Primers were designed based on one Z01-A-specific sequence, A79, and used in a multiplex PCR to discriminate between S. marcescens strains causing CYVD and those from other ecological niches.


Using Machine Learning to Train a Wearable Device for Measuring Students' Cognitive Load during Problem-Solving Activities Based on Electrodermal Activity, Body Temperature, and Heart Rate: Development of a Cognitive Load Tracker for Both Personal and Classroom Use.
Automated tracking of physical fitness has sparked a health revolution by allowing individuals to track their own physical activity and health in real time. This concept is beginning to be applied to tracking of cognitive load. It is well known that activity in the brain can be measured through changes in the body's physiology, but current real-time measures tend to be unimodal and invasive. We therefore propose the concept of a wearable educational fitness (EduFit) tracker. We use machine learning with physiological data to understand how to develop a wearable device that tracks cognitive load accurately in real time. In an initial study, we found that body temperature, skin conductance, and heart rate were able to distinguish between (i) a problem solving activity (high cognitive load), (ii) a leisure activity (moderate cognitive load), and (iii) daydreaming (low cognitive load) with high accuracy in the test dataset. In a second study, we found that these physiological features can be used to predict accurately user-reported mental focus in the test dataset, even when relatively small numbers of training data were used. We explain how these findings inform the development and implementation of a wearable device for temporal tracking and logging a user's learning activities and cognitive load.


Failed epidural anaesthesia in a patient with Hurler's disease.
Failure of epidural anaesthesia in a patient with Hurler's disease is presented. The possibility that epidural anaesthesia may fail due to deposition of mucopolysaccharide in the meninges, or in the epidural space is discussed. A granuloma in the upper trachea and severe choanal stenosis complicated the already difficult airway management.


Transplantation of frozen-thawed mouse primordial follicles.
Primordial follicles were isolated from juvenile mouse ovaries and cryopreserved by slow freezing with dimethylsulphoxide as the cryoprotectant. After thawing, approximately 80% of the oocytes and 65% of the somatic cells excluded Trypan Blue dye, indicating that cell membranes were still intact. Frozen-thawed cells were suspended in plasma clots and transplanted to the ovarian bursas of host animals that had been sterilized by oophorectomy. The grafts of frozen-thawed cells reorganized into morphologically distinguishable ovaries which produced signs of oestrogenic activity. After natural mating, host females produced normal offspring that were demonstrated by genetic markers to be derived from the transplanted frozen-thawed primordial follicles.


Selective Release of Recombinant Periplasmic Protein From E. coli Using Continuous Pulsed Electric Field Treatment.
To date, high-pressure homogenization is the standard method for cell disintegration before the extraction of cytosolic and periplasmic protein from E. coli. Its main drawback, however, is low selectivity and a resulting high load of host cell impurities. Pulsed electric field (PEF) treatment may be used for selective permeabilization of the outer membrane. PEF is a process which is able to generate pores within cell membranes, the so-called electroporation. It can be readily applied to the culture broth in continuous mode, no additional chemicals are needed, heat generation is relatively low, and it is already implemented at industrial scale in the food sector. Yet, studies about PEF-assisted extraction of recombinant protein from bacteria are scarce. In the present study, continuous electroporation was employed to selectively extract recombinant Protein A from the periplasm of E. coli. For this purpose, a specifically designed flow-through PEF treatment chamber was deployed, operated at 1.5 kg/h, using rectangular pulses of 3 μs at specific energy input levels between 10.3 and 241.9 kJ/kg. Energy input was controlled by variation of the electric field strength (28.4-44.8 kV/cm) and pulse repetition frequency (50-1,000 Hz). The effects of the process parameters on cell viability, product release, and host cell protein (HCP), DNA, as well as endotoxin (ET) loads were investigated. It was found that a maximum product release of 89% was achieved with increasing energy input levels. Cell death also gradually increased, with a maximum inactivation of -0.9 log at 241.9 kJ/kg. The conditions resulting in high release efficiencies while keeping impurities low were electric field strengths ≤ 30 kV/cm and frequencies ≥ 825 Hz. In comparison with high-pressure homogenization, PEF treatment resulted in 40% less HCP load, 96% less DNA load, and 43% less ET load. Therefore, PEF treatment can be an efficient alternative to the cell disintegration processes commonly used in downstream processing.


Expression changes in DNA repair enzymes and mitochondrial DNA damage in aging rat lens.
To determine if there is increased mitochondrial DNA (mtDNA) and nuclear DNA (nDNA) damage with age in the lenses of rats. We also explored the immunolocalization of 8-oxoguanine DNA glycosylase 1 (OGG1) and AP endonuclease 1 (APE1) in the lens and studied three of the predominant base excision repair (BER) enzymes: OGG1, APE1, and DNA polymerase gamma (Polgamma). The methods used by this study include the selection of twenty-six male Wistar rats in each group (2 months old and 26 months old) and fourteen male Wistar rats in the 16 months old group. The total DNA of lenses were isolated and the DNA genome was amplified by a long extension-polymerase chain reaction (LX-PCR). We examined mtDNA and nDNA damage with a quantitative polymerase chain reaction (QPCR) assay that was combined with EvaGreen. We also studied the gene expression of mRNA and protein in these key BER enzymes with real time-polymerase chain reaction (RT-PCR) and western blot analysis. There was an increase in oxidative DNA damage, which exists primarily in the mtDNA. The amount of 8-hydroxy-2'-deoxy-guanosine (8-OHdG) in DNA was significantly increased with age. Our experiments demonstrated that the gene expression of mRNA and protein in these key BER enzymes decreased with age. OGG1 and APE1 were localized by immunohistochemistry within lens epithelial cells (LECs) and superficial fiber cells. The gene expression of mRNA and protein in these key BER enzymes decreased with age, which caused a decrease in the repairing capability of the mtDNA and the accumulation of mtDNA damage. The increased mtDNA damage and decreased expression of BER enzymes may cause a "vicious cycle" of oxidative stress that contributes to the accumulation of mtDNA mutations and age-related cataract pathogenesis.


[Effects of curcumin on pneumocyte apoptosis and CHOP in pulmonary ischemia/reperfusion injury of mice].
To investigate the effects of curcumin (CUR) on pneumocyte apoptosis and CCAAT/enhancer binding protein homologous protein (CHOP) in pulmonary ischemia/reperfusion injury (PIRI) in mice. Sixty C57BL/6J mice were randomly allocated into six groups (n = 10): Sham operation group (Sham group), ischemia/reperfusion group (I/R group), ischemia/reperfusion + dimethyl sulfoxide group (DMSO group), ischemia/reperfusion + curcumin pre-treated with respectively 100 mg/kg, 150 mg/kg and 200 mg/kg groups (CUR-100 group, CUR-150 group and CUR-200 group). Left lung tissue of each group was excised after reperfusion for 3 h. Wet lung weight to dry lung weight (W/D) and total lung water content (TLW) were tested. The morphological and ultrastructural changes of lung tissue were observed under light microscope and electron microscope, and index of quantitative evaluation for alveolar damage (IQA) was calculated. The expression levels of CHOP and glucose regulated protein 78 (GRP78) were detected by RT-PCR and Western Blot. Apoptosis index (AI) of lung tissue was determined by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) method. Compared with Sham group, the expression levels of CHOP, GRP78 mRNA and protein were all significantly increased (P < 0.05) in I/R group and DMSO group, W/D, TLW, IQA and AI were all notably higher (P < 0.01); morphological and ultrastructural injury in lung tissue were notably observed in I/R group. Compared with DMSO group, the expression levels of GRP78 mRNA and protein were increased higher (P < 0. 05) in CUR-100 group, CUR-150 group, and CUR-200 group, but the expression levels of CHOP mRNA and protein were decreased lower (P < 0.05), W/D, TLW, IQA and AI were also decreased (P < 0.05, P < 0.01); morphological and ultrastructural injury in lung tissue were gradually alleviated in CUR groups. I/R induces excessive unfolded protein response (UPR) in lung tissue, in which CHOP participates in pneumocyte apoptosis, leading to lung injury; CUR has notable effects on lung protection against I/R injury, which may be related to inhibition of apoptosis mediated by CHOP in excessive UPR.


DOCA-salt induced myocardial sensitization to ventricular fibrillation by isoprenaline in rats. Role of the autonomic nervous system.
The possible role of the autonomic nervous system in the development of myocardial sensitization to isoprenaline produced by DOCA-salt pretreatment was evaluated in bilaterally unanaesthetized rats subject to chronic ganglionic blockade with chlorisondamine or hexamethonium and to treatment with 6-hydroxydopamine. Isoprenaline produced ventricular fibrillation and death in rats pretreated for 6 days with DOCA (20 mg pellet, s.c.) and 0.9% saline. The incidence of mortality was dose dependent and was 76.6% with 150 microgram/kg of isoprenaline. Adrenalectomy did not alter the mortality rate. The combination of adrenalectomy with concurrent chronic ganglionic blockade significantly decreased the incidence of mortality and delayed the time of death. In contrast, 6-hydroxydopamine treatment and adrenalectomy did not alter the incidence of mortality nor time of death. No differences in myocardial noradrenaline turnover were detected in rats susceptible and nonsusceptible to death in ventricular fibrillation. We conclude that myocardial sensitization to isoprenaline induced by DOCA-salt requires a neurogenic component, which is not, however, manifested by changes in myocardial noradrenaline turnover.


TRPM2 expression levels are associated with histological grading in patients with tongue squamous cell carcinoma.
Tongue squamous cell carcinoma (TSCC) accounts for a large proportion of cases of head and neck cancer. Transient receptor potential melastatin 2 (TRPM2) is a non‑selective cation channel sensitive to oxidative stress. High TRPM2 expression has been reported in various types of cancer, including neuroblastoma, glioblastoma, non‑small cell lung cancer and breast cancer. However, whether expression levels of TRPM2 are associated with aggressive clinical features in TSCC remains unclear. A total of 26 clinical sample tissues with TSCC were collected in the present study. The expression levels of the TRPM2 channel were determined by immunohistochemistry, western blot, and qPCR analysis. The content of malondialdehyde (MDA) and the activity of superoxide dismutase (SOD) were measured to reveal oxidative stress levels in TSCC tissues with different degrees of differentiation. The protein expression levels of caspase‑8, Bcl‑xL, Bax, caspase‑9, cleaved caspase‑9, caspase‑3, cleaved caspase‑3, poly [ADP‑ribose] polymerase (PARP) and cleaved PARP were detected by western blot analysis. Analysis of the tissue specimens from 26 patients with TSCC showed that TRPM2 was not upregulated in all specimens. Notably, the expression levels of TRPM2 were associated with the histological grading of different tissues. The specimens with low TRPM2 expression were significantly associated with moderate or poor differentiation (P=0.003), and exhibited increased lipid peroxidation level and decreased SOD activity. Furthermore, the altered expression of pro‑ and anti‑apoptotic proteins indicated a significant upregulation of apoptosis in TSCC tissues with low TRPM2 expression. These results suggested that low TRPM2 expression in TSCC may inhibit the ability of cells to adapt to or resist the oxidative stress, resulting in increased susceptibility to apoptosis. Therefore, the oxidative stress‑sensitive TRPM2 channel may serve as a potent biomarker, and the present study provides insights into the underlying mechanisms of tumor cell differentiation.


[Regenerative medicine with adult stem cells].
The present state of clinical regenerative medicine with adult stem cells in the cardiology, digestive, corneal and neurological fields are reviewed. From the cardiology point of view, there is clinical experience with bone marrow stem cells and peripheral blood cells and with skeletal myoblasts. At present, the adult stem cells (bone marrow hematopoietic or mesenchymal) constitute the best option for the regeneration of heart tissue, the clinical studies showing favorable results without ethical or safety problems. Most of the studies with skeletal myoblasts have also been demonstrated to significantly contribute to improve heart function, above all, the systolic one. However they have the disadvantage that has not been totally clarified that they induce malignant ventricular arrhythmias. In either case, the clinical studies are in the initial phase and new studies, above all randomized, are necessary. In the digestive field, there is the pioneer experience of the Hospital La Paz on the use of stem cells from abdominal fat in the treatment of fistulous condition of patients with Crohn's disease. In ophthalmology, the limbal corneal transplant is a recognized practice, using cells from the contralateral eye when the damage is in a single eye and cells from a donor when the damage is bilateral. Finally, in the neurological field, different zones of the adult mammal brain where there are stem cells have been identified: the hippocampus, subventricular zone, olfactory bulb and periependymal zone of the spinal cord. On the other hand, neurons may be obtained from adult stem cells from other tissues, such as the bone marrow or adipose tissue, which means a practically unendable source of neural precursors, either by direct implant after their selection or after their in vitro culture. However, most of the experimentation is animal up to now, clinical trails on safety in amyotrophic lateral sclerosis are now being initiated.


Irreversible Electroporation: An Emerging Immunomodulatory Therapy on Solid Tumors.
Irreversible electroporation (IRE), a novel non-thermal ablation technique, is utilized to ablate unresectable solid tumors and demonstrates favorable safety and efficacy in the clinic. IRE applies electric pulses to alter the cell transmembrane voltage and causes nanometer-sized membrane defects or pores in the cells, which leads to loss of cell homeostasis and ultimately results in cell death. The major drawbacks of IRE are incomplete ablation and susceptibility to recurrence, which limit its clinical application. Recent studies have shown that IRE promotes the massive release of intracellular concealed tumor antigens that become an "in-situ tumor vaccine," inducing a potential antitumor immune response to kill residual tumor cells after ablation and inhibiting local recurrence and distant metastasis. Therefore, IRE can be regarded as a potential immunomodulatory therapy, and combined with immunotherapy, it can exhibit synergistic treatment effects on malignant tumors, which provides broad application prospects for tumor treatment. This work reviewed the current status of the clinical efficacy of IRE in tumor treatment, summarized the characteristics of local and systemic immune responses induced by IRE in tumor-bearing organisms, and analyzed the specific mechanisms of the IRE-induced immune response. Moreover, we reviewed the current research progress of IRE combined with immunotherapy in the treatment of solid tumors. Based on the findings, we present deficiencies of current preclinical studies of animal models and analyze possible reasons and solutions. We also propose possible demands for clinical research. This review aimed to provide theoretical and practical guidance for the combination of IRE with immunotherapy in the treatment of malignant tumors.


Quercetin-solid lipid nanoparticle-embedded hyaluronic acid functionalized hydrogel for immunomodulation to promote bone reconstruction.
Bone defects are a persistent challenge in clinical practice. Although repair therapies based on tissue-engineered materials, which are known to have a crucial role in defective bone regeneration, have gathered increased attention, the current treatments for massive bone defects have several limitations. In the present study, based on the immunomodulatory inflammatory microenvironment properties of quercetin, we encapsulated quercetin-solid lipid nanoparticles (SLNs) in a hydrogel. Temperature-responsive poly(ε-caprolactone-co-lactide)-b-poly(ethylene glycol)-b-poly(ε-caprolactone-co-lactide) modifications were coupled to the main chain of hyaluronic acid hydrogel, constructing a novel, injectable bone immunomodulatory hydrogel scaffold. Extensive in vitro and in vivo data showed that this bone immunomodulatory scaffold forms an anti-inflammatory microenvironment by decreasing M1 polarization, while elevating the M2 polarization. Synergistic effects on angiogenesis and anti-osteoclastic differentiation were observed. These findings further proved that administering quercetin SLNs encapsulated in a hydrogel can aid bone defect reconstruction in rats, providing new insights for large-scale bone defect repair.


[Biosynthesis of nitric oxide and its function in organism].
Nitric oxide (NO) is a gas with diverse biological activities produced from L-arginine by nitric oxide synthetase (NOS). This article provides a short overview of the basic function of NO which takes part in smooth muscle relaxation, neurotransmission, modulation of inflammation in a number of tissues and pathological conditions. The discovery of these new physiological and pathological pathways allows for better understanding of many complex processes. It opens a new field for better treatment of many diseases.


Visible Light-Driven Photocatalytic Activity of Magnetic Recoverable Ternary ZnFe2O4/rGO/g-C3N4 Nanocomposites.
ZnFe2O4/rGO/g-C3N4 ternary nanocomposite photocatalysts with different ZnFe2O4/g-C3N4 weight ratio (0.5, 0.75, 1) were prepared by a stepwise solvothermal method using ethylene glycol as the solvent. Physicochemical methods such as X-ray diffraction, UV-Vis diffuse reflectance spectroscopy and photoluminescence spectroscopy were applied in order to characterize the composites. The formation of a meso-/macroporous structure with specific surface area between 67 and 77 m2 g?1 was confirmed by N2 adsorption/desorption. The bandgap of the composites was found to be lower (2.30 eV) than that of g-C3N4 (2.7 eV). In contrast to pure g-C3N4, the composites showed no fluorescence, i.e. no recombination of e?/h+ took place. All samples, including pure g-C3N4 and ZnFe2O4, were tested for adsorption and photocatalytic degradation of aqueous malachite green model solutions (10?5 M) under visible light irradiation (? >400 nm). The results show that the prepared nanocomposites have higher absorption and photocatalytic activity than the pristine g-C3N4 and ZnFe2O4 and can be successfully used for water purification from organic azo-dyes.


Convergence and Divergence of Signaling Events in Guard Cells during Stomatal Closure by Plant Hormones or Microbial Elicitors.
Dynamic regulation of stomatal aperture is essential for plants to optimize water use and CO2 uptake. Stomatal opening or closure is accompanied by the modulation of guard cell turgor. Among the events leading to stomatal closure by plant hormones or microbial elicitors, three signaling components stand out as the major converging points. These are reactive oxygen species (ROS), cytosolic free Ca(2+), and ion channels. Once formed, the ROS and free Ca(2+) of guard cells regulate both downstream and upstream events. A major influence of ROS is to increase the levels of NO and cytosolic free Ca(2+) in guard cells. Although the rise in NO is an important event during stomatal closure, the available evidences do not support the description of NO as the point of convergence. The rise in ROS and NO would cause an increase of free Ca(2+) and modulate ion channels, through a network of events, in such a way that the guard cells lose K(+)/Cl(-)/anions. The efflux of these ions decreases the turgor of guard cells and leads to stomatal closure. Thus, ROS, NO, and cytosolic free Ca(2+) act as points of divergence. The other guard cell components, which are modulated during stomatal closure are G-proteins, cytosolic pH, phospholipids, and sphingolipids. However, the current information on the role of these components is not convincing so as to assign them as the points of convergence or divergence. The interrelationships and interactions of ROS, NO, cytosolic pH, and free Ca(2+) are quite complex and need further detailed examination. Our review is an attempt to critically assess the current status of information on guard cells, while emphasizing the convergence and divergence of signaling components during stomatal closure. The existing gaps in our knowledge are identified to stimulate further research.


Differential effects of two hydrocephalus/MASA syndrome-related mutations on the homophilic binding and neuritogenic activities of the cell adhesion molecule L1.
The cell adhesion molecule L1 plays an important role in neural development. We have previously demonstrated that the second immunoglobulin-like domain (Ig2) of L1 contains both homophilic binding and neuritogenic activities (Zhao, X., and Siu, C.-H. (1995) J. Biol. Chem. 270, 29413-29421). Recently, two mutations (R184Q and H210Q) within the Ig2 region of the human L1 gene have been shown to be responsible for X-linked hydrocephalus and the related MASA (mental retardation, aphasia, shuffling gait, and adducted thumbs) syndrome. Glutathione S-transferase-Ig2 fusion proteins containing these mutations were used to evaluate their effects on L1. The homophilic binding activity of fusion proteins and their ability to promote neurite outgrowth from retinal cells were examined. The R184Q mutation led to a complete loss of both homophilic binding and neuritogenic activities, while the H210Q mutation resulted only in a partial loss. These results provide, for the first time, direct demonstration of the deleterious effects of hydrocephalus/MASA mutations on two intrinsic properties of L1.


Artificial Intelligence and the Practice of Neurology in 2035: The Neurology Future Forecasting Series.
High-quality health care delivery relies on a complex orchestration of the flow of patient data. Incorporating advanced artificial intelligence (AI) technologies into this delivery system has tremendous potential to improve health care, but also carries with it unique challenges. The nature of neurologic disease, and the current state of neurologic care delivery, makes this area of medicine well positioned for AI-driven innovation by 2035. Business, ethics, regulation, and medical education will need to evolve in concert. The information technology and data standards requirements for this potential transformation are underappreciated and will be a major driver of changes across the industry. Using AI on patient data to drive health care innovation to improve patients' lives as the primary goal will facilitate widespread acceptance and adoption of the practices required for a successful AI transformation in neurology. In planning the incorporation of AI into clinical practice, the tenets of rigorous research will need to be vigilantly applied to prevent unwarranted costs and inconveniences while promoting meaningful health outcomes.


Nanosized drug-eluting bead for transcatheter arterial chemoembolization (ND-TACE).
Commercially available drug-eluting embolization beads (100-500 μm) reduced the occurrence of adverse events related to an anticancer drug, but were unascertained to remarkably benefit the transcatheter arterial chemoembolization (TACE) treatment of intermediate-stage liver cancer. Dextran-coated arsenite nanoparticles with the size ranging from 400 to 600 nm were developed as a nanosized drug-eluting bead (NDEB) for chemoembolization therapy of the rabbit VX2 liver tumor. We fully characterized their relevant physicochemistry and drug release properties. Their hemolysis was investigated before vessel embolization. The introduction of the NDEB allowed continuous embolization of tumor feeding vessels and sustained release of arsenic trioxide, thereby causing severe tumor necrosis and reduced vascularity. Sonography including B mode ultrasound, color Doppler flow imaging (CDFI) and dynamic contrast-enhanced ultrasound (CEUS) were performed to evaluate the tumor vascularity and viability. Additionally, its hepatotoxicity was tolerable at a medium dose. NDEB-TACE might be an effective therapeutic strategy for interventional therapy.


Detection of a human papilloma virus type 5 DNA in a renal allograft patient from Scotland.
A renal allograft recipient with small raised red scaly lesions on a forearm is described. The clinical and histological picture was that of similar lesions reported in some epidermodysplasia verruciformis patients. Virological investigations revealed the presence of human papilloma virus type 5; this is the second report of the occurrence of this type in allograft patients. As human papilloma virus type 5 is known to have malignant potential, clinicians should be on the lookout for these banal-looking and distinctly non-warty lesions in renal transplant recipients. Close follow-up is indicated.


Changes in quantal parameters of EPSCs in rat CA1 neurones in vitro after the induction of long-term potentiation.
1. Long-term potentiation (LTP) was induced in EPSCs evoked in CA1 pyramidal neurones of young rats in vitro by extracellular stimulation of stratum radiatum. Low frequency stimulation was paired with postsynaptic depolarization to induce LTP, using whole-cell recording techniques. 2. Sufficient control and potentiated records were obtained under stable recording conditions to allow a quantal analysis of eleven EPSCs. The fluctuations in amplitude of all eleven EPSCs were quantized before conditioning stimulation, and they remained quantized after LTP induction, usually with an increased quantal variance. 3. Quantal current was increased by conditioning for nine out of eleven EPSCs. The increase in quantal current was correlated with the percentage increase in the EPSC. For only two EPSCs could the entire potentiation be attributed to an increase in quantal current. 4. The amplitude fluctuations of five control EPSCs could be described by binomial statistics, but after conditioning the binomial description held for only one of these EPSCs. For this EPSC, conditioning caused the release probability to increase from 0.39 +/- 0.05 to 0.47 +/- 0.02. 5. Quantal content was increased by conditioning stimulation for ten out of eleven EPSCs. The increase in quantal content was correlated with the percentage increase in the EPSC. However, for only four EPSCs could the entire potentiation be attributed to an increase in quantal content. 6. Most EPSCs were evoked with a high proportion of response failures. The probability of response failures decreased in eight out of eleven EPSCs following the induction of LTP. There was a negative correlation between the change in the probability of response failures and the amount of LTP. 7. The minimal number of sites at which transmission occurred increased for ten out of eleven EPSCs following LTP induction. Increases in the minimal number of active sites following conditioning were associated with decreases in the probability of response failures for seven out of eleven EPSCs. 8. The induction of LTP usually resulted in changes in the time course of the EPSCs. Cable analysis using a passive compartmental model of a CA1 pyramidal cell suggested that these time course changes were associated with shifts in the average electrotonic location of the active sites following LTP induction, rather than being caused by an increased duration of synaptic current. 9. LTP expression involves postsynaptic modifications to enhance the synaptic current at active sites. New sites are recruited, and our data cannot be used to determine if this is a result of a pre- or a postsynaptic change. Evidence for an increase in release probability was found for one EPSC.


Modulation of B-cell immunoglobulin synthesis by retinoic acid.
Retinoic acid (RA) and its parent compound retinol (ROH, vitamin A) have been recognized as important immunopotentiating agents since the early 1900s. We have focused our studies on the effects of retinoids on B-cell immune function in the newborn infant. The response of cord blood mononuclear cells (CBMC) to formalinized Cowan I strain Staphylococcus aureus (SAC), a T-cell-dependent factor for inducing the differentiation of B cells into immunoglobulin (Ig)-secreting cells, was used as a model system for studying whether RA could alter the immunoglobulin synthesis of newborn B lymphocytes. The addition of RA to SAC-stimulated CBMC cultures produced a 2- to 47-fold increase in IgM synthesis. An ELISA-spot assay showed that the RA-induced enhancement in Ig synthesis was due to the recruitment of more B cells to differentiate into Ig-secreting cells. Whereas RA enhanced IgM production of CBMC stimulated with SAC, RA augmented only IgG production of SAC-stimulated adult peripheral blood mononuclear cells (PBMC). To determine if the differences in dose-response characteristics between CBMC and adult PBMC resided within the target cell, i.e., the B cell, T-cell-enriched and T-cell-depleted (B-cell) fractions from CBMC and adult PBMC were cocultured in various combinations. The isotype, i.e., IgM vs IgG, and the dose-response curve characteristics were intrinsic to the responding B-cell source, i.e., newborn vs adult. Highly purified T cells from CBMC, when preincubated for 36 hr with RA, enhanced IgM synthesis of cord blood B cells. Supernatants from purified T cells generated a factor which could enhance B-cell synthesis. Although interleukin (IL)-2, IL-4, and IL-6 could not be detected by ELISA in the T-cell-derived supernatants, RA probably generates a cytokine/interleukin from T cells which modulates B-cell Ig secretion. RA can also act directly on B cells as evidenced by the augmentation in Ig synthesis of Epstein-Barr virus (EBV)-transformed B-cell lines. These data suggest that RA can have a direct effect on B cells. Since increased proliferation (numbers) of lymphoblastoid B cells was not responsible for the increased amounts of Ig in the supernatant fluids, we examined whether cytokines secreted by EBV-transformed B cells could be acting as an autocrine factor in increasing Ig synthesis. EBV-transformed B-cell clones incubated with RA for 6 days produced a 20- to 45-fold increase in IL-6. An understanding of the mechanisms by which RA enhances B-cell immune function may lead to the use of RA or its derivatives in patients with immune deficiencies and in preterm infants with immature immune systems.


Cyclopia-astomia-agnathia-holoprosencephaly association: a case report.
A female infant is described with cyclopia-astomia-agnathia-holoprosencephaly association. The authors discuss whether the use of salicylates in early pregnancy is implicated.


De Novo Malignancies After Transplantation: Risk and Surveillance Strategies.
De novo malignancies are one of the leading causes of late mortality after liver and kidney transplantation. Nonmelanoma skin cancer is the most common malignancy, followed by posttransplant lymphoproliferative disorder and solid organ tumors. Immunosuppression is a key factor for cancer development, although many other transplant-related and traditional risk factors also play a role. In this review, the authors summarize risk factors and outcomes of frequently encountered de novo malignancies after liver and kidney transplantation to stratify recipients at highest risk. Future efforts in prospectively validated, cost-effective surveillance strategies that improve survival of these complex patients are greatly needed.


Inhibition of deoxyribonuclease activity in the medium surrounding plant protoplasts.
After 1 hour, exogenous deoxyribonucleic acid was degraded within a culture medium at 25 C (pH 6) containing protoplasts of Daucus carota L. var. sativa. Low temperature incubation (1 C) or the addition of 45 millimolar sodium citrate to the medium eliminated DNase activity for at least 4.5 hours. This DNase activity was not reduced at pH 7 or 9, nor by addition of 200 millimolar adenosine 5'-triphosphate.Techniques were developed to ensure high protoplast plating efficiencies and high regenerative capabilities after low temperature treatment and the addition of sodium citrate to the medium. Results indicated citrate concentrations to 45 mm and 1 C temperatures revealed little or no effect on protoplast regeneration capacities. Protoplast viability was 90 to 95% at the time of plating as determined by phenosafranin staining and an estimated 50 to 60% of these undergo cell division in the solid agar medium.


Single-chain precursor of renal gamma-glutamyl transpeptidase.
The two subunits of gamma-glutamyl transpeptidase (EC 22.214.171.124) are derived from a single-chain glycosylated precursor. A small fraction of the propeptide survives proteolytic processing in the rat kidney and has been purified by an immunoaffinity technique. The propeptide contains determinants for both the subunits and its amino acid composition resembles that of the dimeric enzyme. However, the propeptide exhibits less than 2% of the transpeptidase activity shown by the dimeric enzyme.


NG-monomethyl-L-arginine, an inhibitor of nitric oxide synthase, increases extracellular GABA in the striatum of the freely moving rat.
We investigated the effect of an NO synthase inhibitor, NG-monomethyl-L-arginine (L-NMMA), on the levels of endogenous GABA in the rat striatum using in vivo microdialysis. Rats were perfused with the artificial CSF containing L-NMMA (0.1, 0.3 and 1.0 mM) or its inactive isomer D-NMMA (1.0 mM) for 1 h. Infusion of L-NMMA, but not its D-isomer, dose-dependently increased GABA concentration. Co-infusion with tetrodotoxin (1 microM) did not antagonize the increase of GABA induced by L-NMMA. These results show that decreased NO activity enhances GABA release even in the absence of depolarization of GABA neurones. We conclude that NO may be directly acting on GABA nerve terminals and tonically inhibiting GABA release or synthesis under basal conditions.


Characterization of mouse angiogenin-related protein: implications for functional studies on angiogenin.
Angiogenin-related protein (Angrp), the putative product of a recently discovered mouse gene, shares 78% sequence identity with mouse angiogenin (Ang). In the present study, the relationship of Angrp to Ang has been investigated by producing both proteins in bacteria and comparing their functional properties. We find that mouse Ang is potently angiogenic, but Angrp is not, even when assayed at relatively high doses. A deficiency in catalytic capacity, which is essential for the biological activity of Ang, does not appear to underlie Angrp's lack of angiogenicity. In fact, Angrp has somewhat greater ribonucleolytic activity toward tRNA and dinucleotide substrates than does Ang. Instead, an inability to bind cellular receptors is implicated since Angrp does not inhibit Ang-induced angiogenesis. Poor conservation of the Ang receptor recognition sequence 58-69 in Angrp most likely contributes to this defect. However, other substitutions must also influence receptor binding since an Angrp quadruple mutant that is identical to Ang in this segment still lacks both angiogenic activity and the capacity to inhibit Ang. The functional differences between Ang and Angrp, together with evidence presented herein that Angrp is regulated differently than Ang, suggest that the roles of the two proteins in vivo may be quite distinct.


Synthesis of the oxazolidinone fragment of thelepamide.
Thelepamide, an unique ketide-amino acid isolated from a marine annelid worm Thelepus crispus, has a unique oxazolidinone ring derived from cysteine, glycine and valine. Rareness in nature as well as promising bioactive possibility make the oxazolidinone ring an attractive synthetic target. The hydroxy oxazolidinone fragment of thelepamide was prepared by acid-catalysed N,O-acetal formation between a ketoamide and formaldehyde. Lactone-carbonyl selective isopropyl addition to an oxazilidine-dione under Grignard conditions also forms the target compound.


Coexistent partial anodontia and supernumerary tooth in the mandibular arch: a rare case.
Coexistent partial anodontia and supernumerary tooth in the lower jaw is a very uncommon condition. Very few cases have been reported in the literature of this condition, etiology of which is still obscure. Presented here is a rare case of simultaneous presence of single supernumerary tooth together with missing permanent central incisor teeth in the lower jaw without any associated systemic condition or syndrome.


Primary levofloxacin resistance and gyrA/B mutations among Helicobacter pylori in Japan.
Recent years have witnessed a decrease in the rate of Helicobacter pylori eradication due to antimicrobial resistance, clarithromycin or metronidazole resistance in particular. As one of the alternatives to the standard regimens, levofloxacin-containing therapy has been considered a promising regimen. Nevertheless, there is a little information concerning the prevalence of levofloxacin resistance and this resistance mechanism. Levofloxacin susceptibility was examined using E-test in 507 H. pylori strains clinically isolated in Japan from 2001 to 2004. Mutation patterns in the quinolone resistance-determining regions of the gyrA and gyrB genes were evaluated, performing direct sequencing of 68 levofloxacin-resistant and 50 susceptible strains. Primary levofloxacin resistance was found in 76 (15.0%) strains. Fifty-seven (83.8%) of 68 levofloxacin-resistant strains analyzed had point mutations in gyrA at Asn-87 or Asp-91, while seven (14.0%) of 50 susceptible strains had gyrA mutations. There was a significant difference in the occurrence of gyrA mutations between levofloxacin-resistant and -susceptible strains (p < .001). In levofloxacin-resistant strains, the occurrence of gyrA mutations at Asn-87 was most common regardless of minimal inhibitory concentration levels, and that of gyrA mutations at Asp-91 tended to be associated with low-level resistance. A double gyrA mutation at Asn-87 and Asp-91 might have an additional impact. As for gyrB, three (4.4%) of 68 levofloxacin-resistant strains with no susceptible strains had mutations. Primary levofloxacin resistance was common in Japan and primarily related to gyrA mutations at Asn-87 and Asp-91.


Origanum vulgare L. essential oil inhibits virulence patterns of Candida spp. and potentiates the effects of fluconazole and nystatin in vitro.
Recurrence and resistance of Candida spp. infections is associated with the ability of these microorganisms to present several virulence patterns such as morphogenesis, adhesion, and biofilm formation. In the search for agents with antivirulence activity, essential oils could represent a strategy to act against biofilms and to potentiate antifungal drugs. To evaluate the antivirulence effect of Origanum vulgare L. essential oil (O-EO) against Candida spp. and to potentiate the effect of fluconazole and nystatin. The effect of O-EO was evaluated on ATCC reference strains of C. albicans and non-albicans Candida species. Minimum inhibitory concentration (MIC) was determined through broth microdilution assay. Adhesion to microplates was determined by crystal violet (CV) assay. An adapted scratch assay in 24-well was used to determine the effect of essential oil on biofilms proliferation. Viability of biofilms was evaluated by MTT reduction assay and through a checkerboard assay we determined if O-EO could act synergistically with fluconazole and nystatin. MIC for C. albicans ATCC-90029 and ATCC-10231 was 0.01 mg/L and 0.97 mg/L, respectively. For non-albicans Candida strains MIC values were 2.6 mg/L for C. dubliniensis ATCC-CD36 and 5.3 mg/L for C. krusei ATCC-6258. By using these concentrations, O-EO inhibited morphogenesis, adhesion, and proliferation at least by 50% for the strains assayed. In formed biofilms O-EO decreased viability in ATCC 90029 and ATCC 10231 strains (IC50 7.4 and 2.8 mg/L respectively). Finally, we show that O-EO interacted synergistically with fluconazole and nystatin. This study demonstrate that O-EO could be considered to improve the antifungal treatment against Candida spp.


Maternal-fetal conflict: rapidly evolving proteins in the rodent placenta.
Conflicting evolutionary interests between mother and offspring are hypothesized to drive an evolutionary arms race during mammalian pregnancy, and thus, positive selection may cause the rapid divergence of placental proteins that affect maternal or fetal fitness. We investigated the genomic consequences of placental expression in rodents and report that a substantial proportion (20.5%) of genes specifically expressed in the mature placenta are rapidly evolving. Moreover, we found that most rapidly evolving genes belong to just three pregnancy-related gene families: placental cathepsins, prolactins, and placental carcinoembryonic antigens. We then sequenced the most rapidly evolving gene, trophoblast-specific protein alpha (Tpbpa), in nine different Mus species/subspecies and found evidence of positive selection within the Mus lineage, with an excess of nonsynonymous changes clustering near a functionally important interaction site. Together, these results suggest that placental proteins, which mediate interactions between mother and offspring, often may be the targets of evolutionary conflict.


Transcranial direct current stimulation on prefrontal and parietal areas enhances motor imagery.
Considering the potential effect of transcranial direct current stimulation (tDCS) to improve motor imagery the purpose of this study was to investigate the effects of tDCS on prefrontal and postparietal cortex in hand mental rotation (HMR). This investigation was a single-blind, randomized study which 60 healthy right-hand college students (30 males and 30 females, age 24.27±0.19 years) volunteered to attend. Using a simple random method, participants were divided into four groups: anodal: F4 (n=15) and P4 (n=15), sham: F4 (n=15) and P4 (n=15). Participants were asked to perform HMR task before and after five sessions of tDCS. Results showed that there is a significant difference between the pretest and post-test of reaction time (t=10.09, d.f.=29, P=0.005) and accuracy (t=-5.04, d.f.=29, P=0.005) in two sites (F4, P4) in anodal group, also two-way analysis of variance of HMR reaction time showed significant main effect of Group (F=52.458, P=0.000, ηP=0.488) indicating faster response in postanodal Group and Site (F=6.561, P=0.013, ηP=0.107) indicating better response in F4, and in HMR accuracy a significant main effect of Group (F=13.659, P=0.001, ηP=0.199) but not for the main effect of Site (F=0.499, P=0.483, ηP=0.009). According to the findings of the study, it is suggested that tDCS on both prefrontal and postparietal cortex could improve HMR with more effect on prefrontal area.


The Use of Arbuscular Mycorrhizal Fungi to Improve Strawberry Production in Coir Substrate.
Strawberry is an important fruit crop within the UK. To reduce the impact of soil-borne diseases and extend the production season, more than half of the UK strawberry production is now in substrate (predominantly coir) under protection. Substrates such as coir are usually depleted of microbes including arbuscular mycorrhizal fungi (AMF) and consequently the introduction of beneficial microbes is likely to benefit commercial cropping systems. Inoculating strawberry plants in substrate other than coir has been shown to increase plants tolerance to soil-borne pathogens and water stress. We carried out studies to investigate whether AMF could improve strawberry production in coir under low nitrogen input and regulated deficit irrigation. Application of AMF led to an appreciable increase in the size and number of class I fruit, especially under either deficient irrigation or low nitrogen input condition. However, root length colonization by AMF was reduced in strawberry grown in coir compared to soil and Terragreen. Furthermore, the appearance of AMF colonizing strawberry and maize roots grown in coir showed some physical differences from the structure in colonized roots in soil and Terragreen: the colonization structure appeared to be more compact and smaller in coir.


Covert Subclinical Neurocognitive Sequelae During the Rehabilitation Course of Severe Coronavirus Disease 2019.
Apart from respiratory symptoms, encephalopathy and a range of central nervous system complications have been described in coronavirus disease 2019. However, there is a lack of published literature on the rehabilitative course and functional outcomes of severe coronavirus disease 2019 with encephalopathy. In addition, the presence of subclinical neurocognitive sequelae during postacute rehabilitation has not been described and may be underrecognized by rehabilitation providers. We report the rehabilitative course of a middle-aged male patient with severe coronavirus disease 2019 who required intensive care and mechanical ventilation. During postacute inpatient rehabilitation for severe intensive care unit-related weakness, an abnormal cognitive screen prompted brain magnetic resonance imaging, which revealed destructive leukoencephalopathy. Subsequently, detailed psychometric evaluation revealed significant impairments in the domains of processing speed and executive function. After 40 days of intensive inpatient rehabilitation, he was discharged home with independent function. This report highlights the need for an increased awareness of covert subclinical neurocognitive sequelae, the role of comprehensive rehabilitation, and value of routine cognitive screening therein and describes the neurocognitive features in severe COVID-19.


Enantioselective Synthesis of 4-Cyanotetrahydroquinolines via Ni-Catalyzed Hydrocyanation of 1,2-Dihydroquinolines.
A Ni-catalyzed asymmetric hydrocyanation that enables the formation of 4-cyanotetrahydroquinolines in good yields with excellent enantioselectivities is presented herein. A variety of functional groups are well-tolerated, and a gram-scale reaction supports the synthetic potential of the transformation. Additionally, several crucial intermediates for pharmaceutically active agents, including a PGD2 receptor antagonist, are now accessible through asymmetric synthesis using this new protocol.


[Survival and prognosis of patients with polycythemia vera: a study of 816 patients in a single Chinese center].
To explore the survival and the risk factors of poor prognosis in Chinese patients with polycythemia vera (PV). A total of 816 patients with a definite diagnosis of PV were enrolled from August 1983 to June 2013 into this study. The standardized mortality ratio (SMR) was calculated by comparing the cumulative survival of 816 PV patients with age- and sex- and calendar year-matched healthy Chinese population from the national bureau of statistics of the People's Republic of China. The clinical features of diagnosis and prognosis of PV patients were analyzed by Cox regression to identify risk factors for the poor prognosis of PV and to develop a dynamic prognostic model in Chinese patients. The effects of different treatments on the development of acute myelocytic leukemia (AML) and post-PV myelofibrosis (post-PV MF) were determined by Kaplan-Meier analysis. JAK2 V617F allele burden (V617F%) was determined by quantitative real-time PCR in 104 patients. The median follow-up time was 6 (1-42) years. The 10-, 15- and 20-year overall survival (OS) was 89.50%, 76.70% and 64.70%, respectively. The SMR was 17.40 (95% CI: 13.71-21.78). Cox regression analysis revealed that white blood cell (WBC) count>10×10(9)/L (HR=3.10, 95% CI: 1.47-6.53, P=0.003), age>60 years (HR=2.89, 95% CI: 1.84-4.53, P<0.001) and prior thrombosis (HR=2.66, 95% CI: 1.65-4.29, P<0.001) were significant predictors for the poor prognosis of PV. Based on the hazard radio, 816 patents were allocated into 4 categories with significantly different survival: low (sum of points=0; median survival no reached), intermediate 1 (sum of points=1; median survival 33.10 (28.20-38.00) years), intermediate 2 (sum of points=2; median survival 23.00 (16.08-29.92) years), high (sum of points=3; median survival 13.00 (10.58-15.42) years). The mortality of high risk group was 5.37 fold higher than low risk patients. The 10- and 20-year survival of no post-PV MF were 89.50% and 79.60%, respectively, for interferon α (IFN-α); 73.80% and 43.50%, respectively, for hydroxyurea treatment; 82.20% and 71.40%, respectively, for alkylating agent treatment; and 80.00% and 38.20%, respectively, for no cytoreductive treatment. The treatment of exposure to IFN-α associated with a higher rate of no-post-PV MF survival (Log-rank=9.79, P=0.020). There were more post-PV MF patients with V617F%≥50% compared with those V617F%<50% (P<0.001). The mortality of PV patients is significantly higher than that of healthy Chinese population. The WBC count>10×10(9)/L, age>60 years, and prior thrombosis are identified as significant predictors for the prognosis of PV. The risk of post-PV MF transformation may be ameliorated by IFN-α via decreasing the burden of JAK2 V617F mutation.


Survival of fetal hippocampal CA3 cell grafts in the middle-aged and aged hippocampus: effect of host age and deafferentation.
The potential application of neural transplantation to many neurodegenerative disorders at early stages of disease progression would involve middle-aged and aged persons. Hence, it is important to examine critically the extent of graft cell survival in both intact and partially deafferented middle-aged and aged brain. We investigated the degree of survival of 5'-bromodeoxyuridine (BrdU)-labeled fetal hippocampal CA3 cells after grafting into both intact hippocampus and partially deafferented hippocampus (i.e., hippocampus contralateral to intracerebroventricular administration of kainic acid) of middle-aged and aged Fischer 344 rats. Absolute cell survival within these grafts was rigorously analyzed using BrdU immunostaining of serial sections and the optical fractionator cell counting method. In the intact hippocampus, graft cell survival was 23% of injected cells for middle-aged rats and 18% for aged rats, which is consistent with the survival of fetal hippocampal cells in the intact young adult hippocampus reported earlier (Shetty and Turner [1995] Neuroscience 67:561-582). A partial deafferentation at the time of grafting significantly enhanced the degree of graft cell survival to 35% of injected cells in the middle-aged hippocampus and 27% in the aged hippocampus. However, the overall graft cell survival after deafferentation was significantly (30%) greater in the middle-aged hippocampus compared with the aged hippocampus. These results reveal that 1) the degree of survival of fetal neural cells in the intact mature brain remains constant with aging and 2) a partial deafferentation of the mature host brain at the time of grafting enhances survival of grafted fetal cells, regardless of the host age. However, the overall extent of graft cell survival after deafferentation depends on the age of the mature brain at the time of deafferentation.


Right orbitofrontal cortex mediates conscious olfactory perception.
Understanding how the human brain translates sensory impressions into conscious percepts is a key challenge of neuroscience research. Work in this area has overwhelmingly centered on the conscious experience of vision at the exclusion of the other senses--in particular, smell. We hypothesized that the orbitofrontal cortex (OFC) is a central substrate for olfactory conscious experience because of its privileged physiological role in odor processing. Combining functional magnetic resonance imaging, peripheral autonomic recordings, and olfactory psychophysics, we studied a case of complete anosmia (smell loss) in a patient with circumscribed traumatic brain injury to the right OFC. Despite a complete absence of conscious olfaction, the patient exhibited robust "blind smell," as indexed by reliable odor-evoked neural activity in the left OFC and normal autonomic responses to odor hedonics during presentation of stimuli to the left nostril. These data highlight the right OFC's critical role in subserving human olfactory consciousness.


A survey of Daldinia species with large ascospores.
Specimens of Daldinia (Xylariaceae) from around the world possessing large ascospores were studied for teleomorphic and anamorphic morphological characters and compared with authentic material of D. grandis. A culture made from a specimen of D. grandis collected from Ecuador produced stromatic structures, but no conidiogenous structures referable to known xylariaceous anamorphs were observed. D. grandis is reconsidered and viewed as a species of warmer climates in the Americas. Three new species are recognised from new combinations of anamorphic and teleomorphic characters: (1) D. novaezelandiae sp. nov. from New Zealand, a fungus obviously related to D. bakeri, which is also reported from that country; (2) D. dennisii sp. nov., from Australia and New Zealand, of which two varieties are erected; and (3) D. loculatoides sp. nov., from the UK and Canada with affinities to D. loculata. The type of Sphaeria durissima was identified as D. loculata. Notes on further Daldinia spp. are included.


A Case for Hydrogen Sulfide Metabolism as an Oxygen Sensing Mechanism.
The ability to detect oxygen availability is a ubiquitous attribute of aerobic organisms. However, the mechanism(s) that transduce oxygen concentration or availability into appropriate physiological responses is less clear and often controversial. This review will make the case for oxygen-dependent metabolism of hydrogen sulfide (H2S) and polysulfides, collectively referred to as reactive sulfur species (RSS) as a physiologically relevant O2 sensing mechanism. This hypothesis is based on observations that H2S and RSS metabolism is inversely correlated with O2 tension, exogenous H2S elicits physiological responses identical to those produced by hypoxia, factors that affect H2S production or catabolism also affect tissue responses to hypoxia, and that RSS efficiently regulate downstream effectors of the hypoxic response in a manner consistent with a decrease in O2. H2S-mediated O2 sensing is then compared to the more generally accepted reactive oxygen species (ROS) mediated O2 sensing mechanism and a number of reasons are offered to resolve some of the confusion between the two.


Fracture history and risk of breast and endometrial cancer.
Fractures in postmenopausal women may serve as a surrogate measure of bone density, reflecting long-term lower estrogen levels, and lower estrogen levels appear to be inversely associated with breast and endometrial cancer. Breast cancer cases aged 50-79 years (n = 5,559) and endometrial cancer cases aged 40-79 years (n = 739) were enrolled in a US case-control study in 1992-1994 to evaluate the relation between fractures and risk of breast and endometrial cancer. Controls for the breast cancer analysis (n = 5,829) and the endometrial cancer analysis (n = 2,334) were randomly selected from population lists (driver's license and Medicare files). Information on fracture history and other risk factors was obtained by telephone interview. Compared with women without a fracture in the past 5 years, the odds ratios for women with a history of fracture were 0.80 (95% confidence interval (CI): 0.68, 0.94) for breast cancer and 0.59 (95% CI: 0.40, 0.89) for endometrial cancer. Height loss (> or =2.5 cm) and recent fracture history were associated with the lowest risk of breast cancer (odds ratio = 0.62, 95% CI: 0.46, 0.83) and endometrial cancer (odds ratio = 0.15, 95% CI: 0.05, 0.43). These data suggest that the endogenous hormonal factors associated with increased fracture risk are also related to decreased breast cancer risk and, more strongly, to endometrial cancer risk.


Targeting of antigens to B lymphocytes via CD19 as a means for tumor vaccine development.
Ab therapy against surface Ags on tumor cells has demonstrated significant efficacy for some cancers. However, it is costly and patients frequently develop acquired resistance over time. In cases of Ab therapy resistance, T cell responses have been shown to be essential in controlling disease progression. Thus, vaccination that generates a sustained Ab response as well as a T cell response may be more effective and economical. In this article, we have developed a vaccination strategy by targeting protein Ags to B cells via a CD19 single-chain variable fragment miniAb. Using the tumor-associated Ag her-2/neu extracellular domain, we showed that the coengagement of CD19 and BCR induced full B cell activation to produce a high titer of Abs and enhanced CD4 Th2 response and CD8 T cell activation and differentiation. These Abs competitively inhibited humanized her-2/neu Ab binding and were capable of activating the complement and inhibiting human breast cancer growth in vitro. Therapeutic efficacy was demonstrated in vivo using murine mammary carcinoma models. Furthermore, four different extracellular domains of her-2/neu could be targeted to B cells to generate Abs against particular domains with different antitumor properties. This approach may offer a new avenue for vaccine development with significantly lower cost, which may be of use not only for cancer therapy but also for infectious agents.


Development of novel pesticides in the 21st century.
General trends and strategies for novel pesticides are summarized. Global pesticide sales and pesticide discovery research are also briefly reviewed. At least 105 chemical pesticides have been launched during the past decade or are under development: 43 fungicides, 34 insecticides/acaricides, 6 nematicides, 21 herbicides, and 1 herbicide safener. Most of them are safe to humans and environmentally friendly. The most developed fungicides are SDHI (succinate dehydrogenase inhibitors), DMI (demethylation inhibitors), QoI (quinone outside inhibitors), and QiI (quinone inside inhibitors). Due to the development of resistance to fungicides with existing modes of action, many fungicides possessing various novel modes of action have been launched or are under development. The trend of insecticide development is changing from organophosphorus, carbamate, and synthetic pyrethroids to nicotinic and diamide insecticides. During the past decade, compounds possessing a variety of novel modes of action have also been launched or are under development. Flupyradifurone and flupyrimin, exhibiting extremely low honeybee toxicity, have been developed and subjected to practical use. Herbicides possessing varied modes of action, such as acetolactate synthase, p-hydroxyphenylpyruvate dioxygenase, protoporphyrinogen oxidase, and very-long-chain fatty acid elongase inhibition, have been developed, but no herbicides possessing a novel mode action have commercialized in nearly 30 years. It is of interest that cyclopyrimorate, which was recently launched, and tetflupyrolimet, which is under development, have novel modes action: homogentisate solanesyltransferase (HST) and dihydroorotate dehydrogenase (DHODH) inhibition, respectively. The development of useful acaricides and nematicides is also progressing. Some natural product origin pesticides are getting attention.