Secreted leukocyte protease inhibitor is present in aqueous humours from cataracts and other eye pathologies.
Previous studies identified serine, cysteine and metalloproteases in normal aqueous humours (AH) and suggested that a balance between proteases and their inhibitors may play a role in the modulation of the AH outflow. We aimed to determine whether secretory leukocyte protease inhibitor (SLPI), a serine protease inhibitor, is present in AH of patients with cataract and other eye pathologies. AH was collected from 117 cataract patients of which 55 were diagnosed with more when one eye disease: cataract only (n=62), pseudoexfoliation (PEX) (n=26), glaucoma (n=6), diabetes retinopathy (n=4), iritis-uveitis (n=4) and macular degeneration (n=28). The total protein in AH was determined by a Bradford assay and SLPI was analyzed by Western blot and ELISA methods. The average concentration of total protein and SLPI in AH samples was 160+/-15 microg/ml (n=117, +/-SEM) and 500+/-94 pg/ml (n=105), respectively. The cataract patients with additional eye disease(s) showed higher protein levels (201+/-35 microg/ml) than cataract (controls) (128+/-31 microg/ml), P<0.01. It is noteworthy that no correlation was found between SLPI and the total protein concentrations in AH, but SLPI was positively correlated with age (r=0.2, P<0.05). No statistical difference in SLPI levels was found between controls (cataract) and other pathologies, while patients with iritis/uveitis had higher SLPI levels compared to those with diabetes (P<0.05). We show here for the first time that SLPI is present in AH and may play a role as well as serve as a marker in pathological states.


Bloodstream infection caused by Solobacteriummoorei: A case report and literature review.
Solobacterium moorei is an asporogenous, strictly anaerobic, short-to-long bacillus belonging to the family Erysipelotrichidae and phylum Firmicutes. This bacterium rarely causes bloodstream infections. Here, we report a case of the postoperative recurrence and metastasis of gastric antral adenocarcinoma combined with S. moorei bloodstream infection.


Comparison of Femtosecond Laser-Assisted Cataract Surgery and Conventional Phacoemulsification on Endothelial Cell Density When Using Torsional Modality.
To compare the effects of femtosecond laser assisted cataract surgery (FLACS) and manual phacoemulsification on cumulative dissipated energy (CDE), torsional amplitude, and endothelial cell density (ECD). This prospective, randomized study was conducted at Oka Eye Clinic (Fukuoka, Japan). Surgeries were performed using FLACS (with LenSx) or conventional technique in adults ≥20 years with grade 2-4 cataracts. Visits included preoperative, surgery day, and 5 postoperative visits (days 1, 4-10, 20-40, 60-120, and 150-210). Primary endpoint was CDE. Secondary endpoints included ECD percent change at day 150-210 versus preoperative visit and average torsional amplitude on surgery day. Exploratory endpoints included central corneal thickness and corrected distance visual acuity (CDVA). Superiority of FLACS to conventional technique was evaluated using t-tests based on a mixed model for repeated measures. Full analysis set included 53 eyes per group. Mean cataract grade was 2.92±0.58 in FLACS and 2.94±0.57 in conventional group. FLACS versus conventional method had significantly lower mean CDE (0.213±0.334 versus 1.718±0.898%-seconds, respectively; P<0.0001), demonstrating superiority of FLACS. Low endothelial cell loss (ECL) was achieved with both FLACS and conventional methods (1.5±5.6% and 2.7±5.2%; P=0.260). Torsional amplitude was significantly lower for FLACS versus conventional method (19.6±16.0% versus 31.1±6.6%; P<0.0001). Central corneal thickness was comparable for both methods at all visits except day 1; CDVA was comparable for both methods at all postoperative visits. FLACS achieved significantly lower CDE compared with the conventional surgical method (P<0.0001). Low ECL was achieved with both FLACS (1.5%) and conventional (2.7%) methods.


[Significance of characteristics of microorganisms in the pathogenesis B. pyocyaneus infection (an experimental study)].
The experimental Pseudomonas Sepsis was induced by various strains. Biological features of these strains were different and various forms of sepsis were induced. The authors suggested that biological features of microorganisms cause the development of different and various forms of sepsis.


Molt cycle related changes and effect of short term starvation on the biochemical constituents of the blue swimmer crab Portunus pelagicus.
Synthesis and hardening of a new exoskeleton are essential to the arthropod molting process. The present study emphasizes the variations in the levels of hemolymph total free sugars, hepatopancreas glycogen and cuticular proteins during the molting stages of Portunus pelagicus. It also reports the effect of short-term starvation conditions on the biochemical constituents of the hemolymph. Intermolt crabs were subjected to 6 days of starvation and hemolymph samples were taken. Standard biochemical procedures were followed toward the quantification of total proteins, total free sugars and total lipids. The total free sugar level in the hemolymph of P. pelagicus was observed to increase during early premolt D0 (3.108 ± 0.032 g/ml) and a gradual decrease till late postmolt B stage (0.552 ± 0.124 g/ml), suggesting the need for total free sugars to provide energy for the apolysis process. Increase in the levels of hepatopancreas glycogen was observed from 1225 ± 0.04 μg/mg in early premolt D0 to 1700 ± 0.3 μg/mg in late premolt D2-3. This is in correlation with the decreased levels of free sugars during premolt stages, suggesting an increase in the storage of glycogen reserves in the hepatopancreas. Cuticular proteins increased during stage B (2.702 ± 0.093 g/ml) and stage C (3.065 ± 0.012 g/ml), indicating exoskeleton hardening and mineralization. Results of the starvation studies clearly showed a steady decline in the level of total free sugars till day 6 (0.099 ± 0.00 g/ml) when compared to the control (8.646 ± 0.08 g/ml). Gradual decrease of total lipids was also observed from the first day of the experiment (6.088 ± 2.44 g/ml) to the last day of the study (0.401 ± 0.20 g/ml) which was 85% lesser than the control (8.450 ± 0.49 g/ml)suggesting the efficient usage of total sugars to consolidate the loss of energy reserves during starvation. The knowledge of Molt-cycle events can be used as a tool for the evaluation of the developmental state providing a morphological reference system for physiological and biochemical studies related to crab aquaculture. Starvation studies enlightens that increasing carbohydrate levels in crab feed together with good protein content could alleviate the natural effects of starvation, improve farm productivity and reduce the deleterious impact of nitrogen pollution generated by rich-protein feeds used in crab farming.


Effect of temperature on sensory and motor conduction of the rat tail nerves.
To assess the effect of temperature upon conduction velocity, amplitude and signal energy of the sensory and motor rat tail nerves. Sensory and motor responses were recorded from the tail nerves in 10 adult rats at different temperatures, starting from 40 degrees C and cooling down to 16 degrees C in steps of 2 degrees C. The conduction velocity of the various components of the orthodromic sensory response was directly and linearly related to temperature (fastest fibres ranged from 47.7 down to 19.7 m/s), with Q(10) values of approximately 1.30, suggesting that all fibres, regardless of their diameter, were equally sensitive to changes in temperature. The motor conduction was similarly affected with a Q(10) value of 1.28 and a velocity range from 24.2 down to 9.6m/s. Amplitude and energy of the sensory responses were inversely related to temperature, reaching their maximum at 16 degrees C. Energy was by far the most temperature sensitive parameter, with a Q(10) of approximately 3 both for fast or slow conducting fibres. Amplitude and energy of the motor responses also showed an inverse correlation with temperature, but were influenced by a more complex set of factors (neuromuscular synapse, muscle membrane) than the simple neural conduction. Besides providing new normative data upon conduction in the rat tail nerves at different temperatures, our results suggest that this method may represent an excellent tool to study models of peripheral-nerve conduction in vivo under various physiological and pathological conditions.


Action of baclofen, GABA and antagonists on the membrane potential of cultured astrocytes of rat spinal cord.
The action of gamma-aminobutyric acid A (GABAA) and B (GABAB)-agonists has been studied on the membrane potential of astrocytes in explant cultures of rat spinal cord by means of intracellular microelectrode recordings. The GABAB-agonists (-)-baclofen and CGP 27 492 (3-aminopropyl phosphonous acid; 10(-6) to 10(-4) M) caused a hyperpolarization of the majority of astrocytes studied. On approximately 25% of the cells, the compounds had no effect. The hyperpolarization by baclofen (10(-4) M) was reversibly antagonized by the GABAB-antagonist 5-hydroxysaclofen (10(-4) M). GABA and the GABAA-agonist muscimol (10(-4) and 10(-3) M) depolarized approximately two thirds of the glial cells tested, whereas the remaining third remained unaffected. The GABAA-antagonist bicuculline (10(-4) and 10(-3) M) only reduced the depolarization by GABA (10(-4) M) but did not completely block it. On half of the cells tested, the depolarization by GABA was not affected by bicuculline, suggesting that the glial GABAA-receptor is different from the neuronal GABAA-receptor. Our electrophysiological investigations together with recent autoradiographic binding studies strongly suggest the existence of GABAB-receptors on astrocytes whereas there is less evidence for GABAA-sites on these cells.


More interventions do not necessarily improve outcome in critically ill patients.
The organizational structure of health care facilities has been shown to affect outcome in critically ill patients. We evaluated the association between structures, treatments and outcomes in a large cohort of critically ill patients. Prospective multicentre cohort study. A total of 26,186 patients consecutively admitted to 31 intensive care units (ICUs) in Austria from January 1998 through December 2000. The ICUs were divided into three groups according to the size and function of the hospital: community hospitals and specialized trauma centers (group A); central referral hospitals (group B); and teaching hospitals (group C). Group C patients exhibited a significantly higher risk-adjusted mortality (O/E ratio). Although severity of illness at admission in groups B and C was similar, group C patients received significantly more invasive diagnostic and therapeutic interventions throughout their ICU stay: For 7 of 10 invasive interventions identified, odds ratios for group C vs group B patients were significantly increased, even after adjustment for age, gender, severity of illness and reason for admission (odds ratios 1.2-13.1; all 95% CIs >1). Risk-adjusted multivariate analysis confirmed that six of these invasive interventions were independently associated with mortality. Furthermore, nurse-to-patient ratios did not differ between groups, leading to a significantly increased nursing workload in group C ICUs. Several invasive interventions were independently associated with increased mortality. Our results provide strong evidence that this association was responsible in part for the increased risk-adjusted mortality in group C patients.


The calcium-sensing receptor and the hallmarks of cancer.
The calcium-sensing receptor (CaSR) plays a pivotal role in systemic calcium metabolism by regulating parathyroid hormone secretion and urinary calcium excretion. The CaSR is ubiquitously expressed, implying a wide range of functions regulated by this receptor. Abnormal CaSR function affects the development of both calciotropic disorders such as hyperparathyroidism, and non-calciotropic disorders such as cardiovascular disease and cancer, which are the leading causes of mortality worldwide. The CaSR is able to bind a plethora of ligands; it interacts with multiple G protein subtypes, and regulates highly divergent downstream signalling pathways, depending on the cellular context. The CaSR is a key regulator for such diverse processes as hormone secretion, gene expression, inflammation, proliferation, differentiation, and apoptosis. Due to this pleiotropy, the CaSR is able to regulate cell fate and is implicated in the development of many types of benign or malignant tumours of the breast, prostate, parathyroid, and colon. In cancer, the CaSR appears to have paradoxical roles, and depending on the tissue involved, it is able to prevent or promote tumour growth. In tissues like the parathyroid or colon, the CaSR inhibits proliferation and induces terminal differentiation of the cells. Therefore, loss of the receptor, as seen in colorectal or parathyroid tumours, confers malignant potential, suggestive of a tumour suppressor role. In contrast, in prostate and breast tumours the expression of the CaSR is increased and it seems that it favours metastasis to the bone, acting as an oncogene. Deciphering the molecular mechanism driving the CaSR in the different tissues could lead to development of new allosteric drug compounds that selectively target the CaSR and have therapeutic potential for cancer. This article is part of a Special Issue entitled: Calcium and Cell Fate. Guest Editors: Jacques Haiech, Claus Heizmann, Joachim Krebs, Thierry Capiod and Olivier Mignen.


Lantana poisoning of cattle and sheep in New Zealand.
The investigation of a field case of Lantana camara poisoning in cattle is reported. Toxicity was confirmed by the experimental dosing of two sheep with dried leaf powder at 6 and 12 mg/kg body weight. Marked cholestatic hepatophy and renal tubular disease resulted. This is the first reported case of toxicity due to grazing a Lantana camara variety grown in New Zealand.


Targeting of integrin-linked kinase with a small interfering RNA suppresses progression of experimental proliferative vitreoretinopathy.
Integrin-linked kinase (ILK) is a serine/threonine kinase that interacts through its COOH terminus with beta1 and beta3 integrins, which mediates a diversity of cell functions by coupling integrins and growth factors to cascades of downstream signaling events. The purpose of this work was to investigate the effects of ILK on development of experimental proliferative vitreoretinopathy (PVR). Cultured human RPE cell line D407 was knocked down for ILK using a small interfering RNA (siRNA). For this, cellular ILK expression was quantified by real-time quantitative PCR, Western blot analysis and immunocytochemical assay, and cytotoxicity of transfection was determined by MTT assay. Moreover, cell attachment, spreading, migration, microfilament dynamics, and cell cycling assays were performed. Furthermore, the impact of the ILK-specific siRNA on PVR was tested using a rabbit model in which PVR was induced by the injection of human RPE cells. Prevalence of PVR and retinal detachment were determined by indirect ophthalmoscopy on days 1, 3, 7, 14, 21 and 28 post-injection. The results showed that blocking the expression of ILK by siRNA significantly inhibited human RPE cell attachment, spreading, migration and proliferation. The knockdown of ILK also disturbed F-actin assembly and induced a cellular arrest in the G1 phase of the cell cycle. Though the eyes injected with ILK-specific siRNA also developed features of PVR, the severities of day 28 post-injection were significantly lower than those in the control eyes (P<0.01). We conclude that targeting of ILK with a small interfering RNA not only inhibits human RPE cell attachment, spreading, migration and proliferation in vitro, but also effectively suppresses development of proliferative vitreoretinopathy in a rabbit model. This may be a potential therapeutic usefulness in treating PVR.


The book of heat from Rashīd ad-Dīn's Kitāb Bayān al-Haqā'iq, MS. Klç Ali Pasha 834, fol. 208 b-241 b Chinese medicine from a source in 14th Century Persia.
Rashid ad-Din gives a summary of what he found in Arabic and Chinese medical literature concerning the topic Heat.


How actin initiates the motor activity of Myosin.
Fundamental to cellular processes are directional movements driven by molecular motors. A common theme for these and other molecular machines driven by ATP is that controlled release of hydrolysis products is essential for using the chemical energy efficiently. Mechanochemical transduction by myosin motors on actin is coupled to unknown structural changes that result in the sequential release of inorganic phosphate (Pi) and MgADP. We present here a myosin structure possessing an actin-binding interface and a tunnel (back door) that creates an escape route for Pi with a minimal rotation of the myosin lever arm that drives movements. We propose that this state represents the beginning of the powerstroke on actin and that Pi translocation from the nucleotide pocket triggered by actin binding initiates myosin force generation. This elucidates how actin initiates force generation and movement and may represent a strategy common to many molecular machines.


Time to loading and locoregional control in perioperative high-dose-rate brachytherapy: The tumor bed effect revisited.
To determine whether the time to loading (TTL) affects locoregional control. Locoregional control status was determined in 301 patients enrolled in several perioperative high-dose-rate brachytherapy (PHDRB) prospective studies conducted at the University of Navarre. The impact of the time elapsed from catheter implantation to the first PHDRB treatment (TTL) was analyzed. Patients treated with PHDRB alone (n = 113), mainly because of prior irradiation, received 32 Gy in eight twice-a-day treatments or 40 Gy in 10 twice-a-day treatments for negative or close/positive margins, respectively. Patients treated with PHDRB + external beam radiation therapy (EBRT) (n = 188) received 16 Gy in four twice-a-day treatments or 24 Gy in six twice-a-day treatments for negative or close/positive margins followed by 45 Gy of EBRT in 25 treatments. After a median followup of 6.5 years (range, 2-13.6+), 113 patients have failed (37.5%), 65 in the PHDRB-alone group (57.5%) and 48 in the combined PHDRB + EBRT group (25.5%). Patients who started PHDRB before Postoperative Day 5 had a 10-year locoregional control rate of 66.7% and patients who started PHDRB on Postoperative Day 5 or longer had a 10-year locoregional control rate of 51.8% (p = 0.009). Subgroup analysis detected that this difference was only observed in the recurrent cases treated with PHDRB alone (Subset 2; n = 99; p = 0.004). No correlation could be detected between locoregional control rate and TTL in the other patient subsets although a trend toward a decreased locoregional control rate after a longer TTL was observed when they were grouped together (p = 0.089). Patients should start PHDRB as soon as possible to maximize locoregional control especially in those recurrent cases treated with PHDRB alone. The time effect in other disease scenarios is less clear.


Clearly weighing the evidence in read-across can improve assessments of data-poor chemicals.
This paper provides a systematic weight-of-evidence method for read-across analyses of data-poor chemicals. The read-across technique extrapolates toxicity from analogous chemicals for which suitable test data are available to a target chemical. To determine that a candidate analogue is the 'best' and is sufficiently similar, the evidence for similarity of each candidate analogue to the target is weighed. We present a systematic weight of evidence method that provides transparency and imposes a consistent and rigorous inferential process. The method assembles relevant information concerning structure, physicochemical attributes, toxicokinetics, and toxicodynamics of the target and analogues. The information is then organized by evidence types and subtypes and weighted in terms of properties: relevance, strength, and reliability into weight levels, expressed as symbols. After evidence types are weighted, the bodies of evidence are weighted for collective properties: number, diversity, and coherence. Finally, the weights for the types and bodies of evidence are weighed for each analogue, and, if the overall weight of evidence is sufficient for one or more analogues, the analogue with the greatest weight is used to estimate the endpoint effect. We illustrate this WoE approach with a read-across analysis for screening the organochlorine contaminant, p,p'-dichlorodiphenyldichloroethane (DDD), for noncancer oral toxicity.


The efficacy of pegylated interferon alpha 2a or 2b plus ribavirin in chronic hepatitis C patients.
We aimed to compare viral responses to pegylated interferon 2a plus ribavirin with pegylated interferon alpha 2b plus ribavirin. Patients with the following characteristics were included: anti HCV(+); normal and/or elevated serum transaminase levels; positive HCV RNA by quantitative PCR; and at least stage 1 fibrosis according to Knodell Scoring System on liver biopsy. Patients were assigned into two groups. Group 1 consisted of 37 patients (24 female, 13 male) who received pegylated interferon alpha 2a 180 microg s.c. weekly plus ribavirin adjusted for patient's weight. All patients were genotype 1. Group 2 consisted of 37 patients (27 female, 10 male) who received pegylated interferon alpha 2b 1.5 microg/kg s.c. weekly plus ribavirin adjusted for patient's weight. At week 24, the treatment was discontinued in patients positive for HCV RNA by PCR, while patients negative for HCV RNA continued treatment up to 48 weeks. The end of treatment and sustained virologic responses of the patients were ascertained by assessing HCV RNA levels at the end of the treatment and after 24 weeks follow-up after the cessation of treatment. At week 48, the proportion of patients with negative HCV RNA (end of treatment viral response) was 28/37 (75.7%) in Group 1 and 27/37 (73%) in Group 2. The group sustained virologic response rates were 48.6% and 35.1% for Group 1 and Group 2, respectively. No significant differences were noted between the two groups. The two pegylated interferon molecules were similar in terms of sustained virologic response rate.


Clopidogrel: new indication. In combination with aspirin: marginal additional benefits.
In patients with myocardial infarction who are not eligible for angioplasty, adding clopidogrel to aspirin reduces the overall 15-day mortality rate, but the subsequent outcome is not known.


On identification of the number of best treatments using the Newman-Keuls test.
In this paper, we provide a stochastic ordering of the Studentized range statistics under a balanced one-way ANOVA model. Based on this result we show that, when restricted to the multiple comparisons with the best, the Newman-Keuls (NK) procedure strongly controls experimentwise error rate for a sequence of null hypotheses regarding the number of largest treatment means. In other words, the NK procedure provides an upper confidence bound for the number of best treatments.


Exploring Minimally Invasive Approach to Define Stages of Type 1 Diabetes Remotely.
Objective: New methods are pivotal in accurately predicting, monitoring, and diagnosing the clinical manifestation of type 1 diabetes (T1D) in high-risk children. Continuous glucose monitoring (CGM) is a valuable tool for patients with T1D, but there is still a knowledge gap regarding its utility in the prediction of diabetes. The current study explored whether 10-day CGM or CGM during an oral glucose tolerance test (OGTT) performed in the laboratory or at home (home-OGTT) could be accurate in detecting stages of T1D. Research Design and Methods: Forty-six subjects 4-25 years of age carrying genetic risk for T1D were recruited and classified into the following groups: islet autoantibody (IAb) negative, one IAb, and stages 1-3 of T1D, based on the laboratory OGTT and IAb results at baseline. A 10-day CGM was initiated before the OGTT. Results: In this study, we showed that CGM was sensitive in detecting asymptomatic individuals at stage 3, and dysglycemic individuals in stage 2 of T1D both during OGTT and the 10-day period. CGM also showed significant differences in several variables during the 10-day sensoring among individuals at different stages of T1D. Furthermore, CGM showed different OGTT profiles and detected significantly more abnormal OGTT results when compared with plasma glucose. Conclusions: CGM together with home-OGTT could detect stages of T1D and offer an alternative method to confirm normoglycemia in high-risk individuals.


A new technique for measuring muscle fiber conduction velocities in full interference patterns.
The motor unit potential shape, mainly its duration and frequency spectra, and the EMG IP crispiness and its frequency spectra are affected by the muscle fiber conduction velocities (MFCVs). Present techniques are somewhat deficient in that they are not adaptable to measure MFCVs continuously and intramuscularly in the presence of interference patterns, and to do so without interfering with the ongoing muscular activity. In this study a cross-correlation with averaging correlograms technique is presented. An EMG needle electrode, with two recording surfaces 1 cm apart, continuously record two channels of EMG activity which is analog-to-digital converted. Contiguous segments of the signals are cross-correlated, the evolved correlograms are averaged together, averaging-out the time-unlocked noise, and averaging-in a peak that represent the average time it takes the EMG signal to propagate from one recording surface to the other. From the distance between these two recording surfaces and the above calculated propagation time the MFCVs can be computed and monitored intramuscularly either in weak or in strong, in isometric or isotonic contractions. But for the fact that a needle is introduced, there is no interference with the muscle electrical activity. It is expected that this technique may add to EMG diagnosis of neuromuscular disorders, will be used to monitor muscular fatigue and applied in normalizing EMG spectra, conditioning them for a better use in diagnostic electromyography.


An aberrant branch of the radial digital nerve in the index finger: case report.
This report describes an incidental finding of an aberrant branch of the radial digital nerve in the index finger of a 39-year-old woman who cut herself with a knife.


P38 mitogen activated protein kinase is involved in the downregulation of granulocyte CXC chemokine receptors 1 and 2 during human endotoxemia.
Chemokine receptors CXC receptor (CXCR) 1 and 2, and their ligands interleukin (IL)-8 and growth-related oncogene alpha (GRO alpha), are principal regulators of neutrophil activation and migration. To investigate the role of p38 mitogen activated protein kinase (MAPK) in the regulation of CXCR expression during an inflammatory response in vivo, 24 healthy volunteers received an intravenous injection with lipopolysaccharide (LPS) preceded (-3 hr) by a specific p38 MAPK inhibitor (BIRB 796 BS) at a high dose (600 mg) or a low dose (50 mg) or a placebo. The LPS-induced reduction of neutrophil CXCR 1 and 2 expression, as determined by fluorescence-activated cell sorter analysis, was inhibited in volunteers receiving the high dose of the p38 MAPK inhibitor. The kinase inhibitor also dose dependently diminished the LPS-induced rises in plasma IL-8 and GRO alpha levels. These results indicate a principal role for p38 MAPK in regulating factors essential for neutrophil activation and chemotaxis in vivo.


Avirulent K88 (F4)+ Escherichia coli strains constructed to express modified enterotoxins protect young piglets from challenge with a virulent enterotoxigenic Escherichia coli strain that expresses the same adhesion and enterotoxins.
Virulence of enterotoxigenic Escherichia coli (ETEC) is associated with fimbrial adhesins and enterotoxins such as heat-labile (LT) and/or heat-stable (ST) enterotoxins. Previous studies using a cell culture model suggest that exclusion of ETEC from attachment to epithelial cells requires expression of both an adhesin such as K88 (F4) fimbriae, and LT. To test the ability of non-pathogenic E. coli constructs to exclude virulent ETEC sufficiently to prevent clinical disease, we utilized a piglet ETEC challenge model. Thirty-nine 5-day-old piglets were inoculated with a placebo (control), or with either of the three K88(+)E. coli strains isogenic with regard to modified LT expression: 8017 (pBR322 plasmid vector control), non-toxigenic mutant 8221 (LT(R192G)) in pBR322, or 8488, with the LT gene fused to the STb gene in pBR322 (LT(R192G)-STb). Piglets were challenged with virulent ETEC Strain 3030-2 (K88(+)/LT/STb) 24h post-inoculation. K88ac receptor-positive piglets in the control group developed diarrhea and became dehydrated 12-24h post-challenge. Piglets inoculated with 8221 or 8488 did not exhibit clinical signs of ETEC disease; most piglets inoculated with 8017 showed diarrhea. Control pigs exhibited significant weight loss, increased blood total protein, and higher numbers of colony-forming units of 3030-2 E. coli in washed ileum and jejunum than treated pigs. This study shows for the first time that pre-inoculation with an avirulent strain expressing adhesive fimbriae and a non-toxic form of LT provides significant short term protection from challenge with a virulent ETEC strain that expresses the same fimbrial adhesion and enterotoxin.


Implementing a Standardized Cardiac Protocol for the Adolescent Preparticipation Sports Examination.
To evaluate adolescent athletes for cardiovascular (CV) abnormalities before engaging in competitive sports, the American Heart Association guidelines recommend preparticipation screening. The initiative aimed to implement a standardized protocol to increase the identification of CV abnormalities during athletic physical examinations. A rapid-cycle model with four Plan-Do-Study-Act cycles was implemented in two urgent care clinics. Providers used a CV-specific screening checklist; parents were engaged in a discussion about the importance of this screening, and team engagement activities were used. Outcomes were measured with chart audits and surveys. Because of the intervention, 62% of athletes were screened, with 29% identified as having a CV risk. The satisfaction of the patients and the team increased by 3% and 62.5%, respectively. Despite competing foci in the clinics, athletes were properly screened using a standardized checklist, which is easily sustainable and can be duplicated in other settings.


Autophagy regulates trophoblast invasion by targeting NF-κB activity.
Preeclampsia is one of the most serious complications of pregnancy, affecting 5-10% of parturients worldwide. Recent studies have suggested that autophagy is involved in trophoblast invasion and may be associated with defective placentation underlying preeclampsia. We thus aimed to understand the mechanistic link between autophagy and trophoblast invasion. Using the two most commonly used trophoblast cell lines, JEG-3 and HTR-8/SVneo, we inhibited autophagy by ATG5 and beclin-1 shRNA. Conversion of LC3-II was evaluated in ATG5 and beclin-1 knock-down cells in the presence of the lysosomal protease inhibitors E-64d and pepstatin A, to detect the efficiency of autophagy inhibition. Upon autophagy inhibition, we measured cell invasion, activity of NF-κB and related signaling pathways, MMP-2, MMP-9, sFlt-1, and TNF-α levels. Autophagy inhibition increased the invasiveness of these trophoblastic cell lines and increased Akt and NF-κB activity as well as p65 expression. Of note, an NF-κB inhibitor significantly attenuated the trophoblast invasion induced by autophagy inhibition. Autophagy inhibition was also associated with increased MMP-2 and MMP-9 levels and decreased the production of sFlt-1 and TNF-α. Collectively, our results indicate that autophagy regulates trophoblast invasiveness in which the NF-κB pathway and MMP-2, MMP-9, sFlt-1 and TNF-α levels are affected.


Drug release in vivo and efficacy evaluation of a new colon targeted powder of total saponins of Pulsatilla.
Based on the anti-ulcerative colitis (UC) effect of total saponins of Pulsatilla (PTS), a pH dependent colonic targeting particle design powder of PTS was prepared. The core-shell composite particle design powder of PTS was prepared with pH sensitive polymer material Eudragit S100 superfine powder as shell and the drug as core. The release of PTS composite particle design powder was increased in the artificial colon fluid and decreased in the artificial stomach and small intestine fluid. In this paper, the release performance of Pulsatilla saponin D in PTS and the ulcerative colitis model induced by TNBS in rats were used to evaluate the targeting of PTS composite particle design powder in colon. The results showed that the content of Pulsatilla saponin D in colon tissue was significantly higher than that of the original drug group after oral administration of PTS composite particle design powder. The solubility of Pulsatilla saponin D in colon tissue was also higher than that in the stomach and small intestine. The peak time and retention time in vivo were prolonged, and the maximum blood concentration was decreased (Cmax). The effect of colonic targeting powder of PTS (50 mg/kg)on anti-ulcerative colitis induced by TNBS in SD rats was better than the original drug (200 mg/kg). Therefore, it is a great significance to make the PTS into colon targeted preparation for improving bioavailability, efficacy and reducing gastrointestinal stimulation.


A novel polysaccharide from Lentinus edodes Mycelia exhibits potential antitumor activity on laryngeal squamous cancer cell line Hep-2.
A novel polysaccharide [Lentinus edodes mycelia polysaccharide 2 (LMP2)] with a molecular weight of 2.27 × 10(4) Da was isolated from the Lentinus edodes mycelia and purified by Sephadex G-200 and diethylaminoethyl-32 column chromatography. HPLC results indicated that LMP2 contained mannose, arabinose, galactose, xylose, and rhamnose with a relative molar ratio of 1:0.74:3.23:1.18:10.98. Its antitumor activity was evaluated in vitro by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, colony formation assay, and transwell assay. LMP2 showed a remarkable inhibitory effect on the proliferation of Hep-2 cells, and at the concentration of 200 mg/ml, the inhibition ratio was 37.2 % after 72 h. Furthermore, colony formation of Hep-2 cells was reduced significantly after treatment with LMP2. Invasion of Hep-2 cells was inhibited significantly by LMP2. These results suggested that LMP2 could be explored as a potential antitumor material for laryngeal carcinoma.


Volumetric evaluation of the relations among the cerebrum, cerebellum and brain stem in young subjects: a combination of stereology and magnetic resonance imaging.
The Cavalieri estimator using a point grid is used to estimate the volume of three-dimensional structures based on two-dimensional slices of the object. The size of the components of intracranial neural structures should have proportional relations among them. The volume fraction approach of stereological methods provides information about volumetric relations of the components of structures. The purpose of our study is to estimate the volume and volume fraction data related to the cerebrum, cerebellum and brain stem. In this study, volume of the total brain, cerebrum, cerebellum and brain stem were estimated in 24 young Turkish volunteers (12 males and 12 females) who are free of any neurological symptoms and signs. The volume and volume fraction of the total brain, cerebrum, cerebellum and brain stem were determined on magnetic resonance (MR) images using the point-counting approach of stereological methods. The mean (+/-SD) total brain, cerebrum and cerebellum volumes were 1,202.05 +/- 103.51, 1,143.65 +/- 106.25 cm3 in males and females, 1,060.0 +/- 94.6, 1,008.9 +/- 104.3 cm3 in males and females, 117.75 +/- 10.7, 111.83 +/- 8.0 cm3 in males and females, respectively. The mean brain stem volumes were 24.3 +/- 2.89, 22.9 +/- 4.49 cm3 in males and females, respectively. Our results revealed that female subjects have less cerebral, cerebellar and brain stem volumes compared to males, although there was no statistically significant difference between genders (P > 0.05). The volume ratio of the cerebrum to total brain volume (TBV), cerebellum to TBV and brain stem to TBV were 88.16 and 88.13% in males and females, 9.8 and 9.8% in males and females, 2.03 and 2.03% in males and females, respectively. The volume ratio of the cerebellum to cerebrum, brain stem to cerebrum and brain stem to cerebellum were 11.12 and 11.16% in males and females, 2.30 and 2.31% in males and females, 20.7 and 20.6% in males and females, respectively. The difference between the genders was not statistically significant (P > 0.05). Our results revealed that the volumetric composition of the cerebrum, cerebellum and brain stem does not show sexual dimorphism.


FunAndes - A functional trait database of Andean plants.
We introduce the FunAndes database, a compilation of functional trait data for the Andean flora spanning six countries. FunAndes contains data on 24 traits across 2,694 taxa, for a total of 105,466 entries. The database features plant-morphological attributes including growth form, and leaf, stem, and wood traits measured at the species or individual level, together with geographic metadata (i.e., coordinates and elevation). FunAndes follows the field names, trait descriptions and units of measurement of the TRY database. It is currently available in open access in the FIGSHARE data repository, and will be part of TRY's next release. Open access trait data from Andean plants will contribute to ecological research in the region, the most species rich terrestrial biodiversity hotspot.


Conformational transition pathway in the allosteric process of calcium-induced recoverin: molecular dynamics simulations.
Recoverin is an important neuronal calcium sensor (NCS) protein, which have been implicated in a wide range of Ca(2+) signaling events in neurons and photoreceptors. To characterize the conformational transition of recoverin from the myristoyl sequestered state to the extrusion state, a series of conventional molecular dynamics (CMD) and targeted molecular dynamics (TMD) simulations were performed. The 36.8 ns long CMD and TMD simulations on recoverin revealed a reliably conformational transition pathway, which can be viewed as a sequential two-stage process. A very important mechanistic conclusion from the present TMD simulations is that the hydrophobic and hydrophilic interactions modulate the allostery cooperatively in the conformational transition pathway. In the first stage, three salt-bridges broken between Lys-84 and Gly-124, between Lys-5 and Glu-103 and between Gly-16 and Lys-97 are major components to destabilize the structure of state T and trigger the swivel of the N- and C-terminal domains. In the second stage, the rupture of H-bond Phe-56-O(...)H(O)-Thr-21 leads to the two helices of EF-1 apart from each other, destabilizing the hydrophobic interactions of the myristoyl group with its environment, whereas the making of H-bond Leu-108-O(...)H(O)-Ser-72 helps the interfacial domain maintain its structural integrity during the course of the myristoyl extrusion. The molecular dynamics simulations results are beneficial to understanding the mechanism of how and why NCS proteins make progress in the photo-signal transduction processes. Further experimental and theoretical studies are still very desirable.


The cap-assisted technique enhances colonoscopy training: prospective randomized study of six trainees.
Colonoscopy and polypectomy procedures have effectively reduced the incidence of colorectal cancer. Currently, competence in colonoscopy is an essential part of the education program for gastrointestinal (GI) trainees. However, considerable training is required for the optimal performance of a colonoscopy. This study involved six colonoscopy trainees, three of whom used the cap whereas the others did not. Each trainee managed 100 cases of screening colonoscopy from beginning to end. The cecal intubation success rate, cecal intubation time, polyp detection rate, adenoma detection rate, advanced adenoma detection rate, and adenocarcinoma detection rate were checked. The rate of successful cecal intubation and the cecal intubation time were reviewed every 10 cases. The cecal intubation rate was 80.7 % (242/300) in the cap group and 63.3 % (190/300) in the non-cap group. The average cecal intubation time was 13.7 min in the cap group and 18.7 min in the non-cap group. The statistical analysis of these results suggested that the cap group had a significantly higher success rate (p < 0.001) and a shorter cecal intubation time (p < 0.001) than the non-cap group. However, the two groups did not differ significantly in the detection rate for polyps (45.3 vs 43 %; p = 0.565), adenomas (26.3 vs 25 %; p = 0.709), advanced adenomas (2.6 vs 0.6 %; p = 0.056), or adenocarcinomas (5.3 vs 3 %; p = 0.153). Cap-assisted colonoscopies might help to increase the rate of cecal intubation success and shorten the cecal intubation time for GI trainees.


Dimethyl-10,12-benz(a)acridine; evidence for differential effects on the synthesis of RNA of mammalian or avian fibroblasts and some RNA viruses.
We have studied the differential effect of dimethyl-10,12-benz(a)acridine (DBMAcr) on the synthesis of RNA of chicken or mouse fibroblasts in culture and that of some RNA-containing viruses such as Rous sarcoma virus and Mengovirus. DMBAcr at low concentrations blocks the cell multiplication of both normal and Rous sarcoma virus-transformed chicken fibroblasts in culture; it affects transformed cells more than normal ones. The cell growth inhibiting effect of DMBAcr is reversible after short periods of incubation. DMBAcr depresses the synthesis of cellular DNA and RNA in parallel. Concurrently the synthesis of protein proceedes at a relatively high rate in DMBAcr-treated cultures. Its inhibitory effect on cellular RNA synthesis is mostly due to a block in the formation of 28 S and 18 S ribosomal RNA species; in contrast, the synthesis of 45 S ribosomal RNA precursor is proceeding at almost control rate. Also, the synthesis of heterogeneous nuclear RNA is not blocked by DMBAcr. The production of Rous sarcoma virus in transformed fibroblasts is not affected by DMBAcr. Since this is correlated with persisting high rates of protein and heterogenous nuclear RNA synthesis, the effects of DMBAcr suggest that the synthesis of Rous sarcoma virus-RNA shares the specificity of messenger and heterogeneous nuclear RNA. DMBAcr inhibits the synthesis of viral RNA of Mengovirus under conditions where the synthesis of total cellular RNA is not appreciably depressed, suggesting its differential effect on the DNA-directed and the RNA-directed RNA synthesis.


Neglected diseases in the news: a content analysis of recent international media coverage focussing on leishmaniasis and trypanosomiasis.
Although the pharmaceutical industry's "neglect" of neglected tropical diseases (NTDs) has been investigated, no study evaluating media coverage of NTDs has been published. Poor media coverage exacerbates the neglect. This study aimed to investigate, describe, and analyse international media coverage of "neglected diseases" in general and three specific NTDs--African trypanosomiasis, leishmaniasis, and Chagas disease--from 1 January 2003 to 1 June 2007. Archives of 11 leading international, English-language media were searched. A content analysis was done, coding for media organisation, date, author, type of report, slant, themes, and "frames". Semi-structured interviews with journalists and key informants were conducted for further insight. Only 113 articles in a 53-month time period met the inclusion criteria, with no strong trends or increases in coverage. Overall, the BBC had the highest coverage with 20 results, followed by the Financial Times and Agence France Presse. CNN had the least coverage with one result. The term "neglected diseases" had good media currency and "sleeping sickness" was far more widely used than trypanosomiasis. The disease most covered was leishmaniasis and the least covered was Chagas. Academic researchers were most commonly quoted as a main source, while the World Health Organization (WHO) and pharmaceutical industry were the least quoted. Journalists generally agreed NTDs had not been adequately covered, but said a lack of real news development and the need to cater to domestic audiences were major obstacles for NTD reporting. All journalists said health agencies, particularly WHO, were not communicating adequately about the burden of NTDs. Public health agencies need to raise priority for NTD advocacy. Innovative strategies, such as reporting grants or creating a network of voices, may be needed.


Titanium-coated hollow screw and reconstruction plate system (THORP) in mandibular reconstruction.
Clinical results are presented which have been obtained from mandibular reconstruction with the Titanium-coated Hollow Screw and Reconstruction-Plate system (THORP) in 7 selected patients. In 3 of the patients primary bone grafting was performed with simultaneous implantation of titanium fixtures for dental reconstruction. The reconstruction-plates have shown excellent functional stability with no failures, such as loosening of screws, within an observation period up to 2 1/2 years. The system was submitted to immediate full functional loading despite the limited number of 2-3 fixation screws for each end of the plate. Among several technical advantages of the system are the possibility of lingual plate application and reconstruction with a three-dimensionally free, adjustable condylar prosthesis.


Neural stem cells for disease modeling and evaluation of therapeutics for Tay-Sachs disease.
Tay-Sachs disease (TSD) is a rare neurodegenerative disorder caused by autosomal recessive mutations in the HEXA gene on chromosome 15 that encodes β-hexosaminidase. Deficiency in HEXA results in accumulation of GM2 ganglioside, a glycosphingolipid, in lysosomes. Currently, there is no effective treatment for TSD. We generated induced pluripotent stem cells (iPSCs) from two TSD patient dermal fibroblast lines and further differentiated them into neural stem cells (NSCs). The TSD neural stem cells exhibited a disease phenotype of lysosomal lipid accumulation. The Tay-Sachs disease NSCs were then used to evaluate the therapeutic effects of enzyme replacement therapy (ERT) with recombinant human Hex A protein and two small molecular compounds: hydroxypropyl-β-cyclodextrin (HPβCD) and δ-tocopherol. Using this disease model, we observed reduction of lipid accumulation by employing enzyme replacement therapy as well as by the use of HPβCD and δ-tocopherol. Our results demonstrate that the Tay-Sachs disease NSCs possess the characteristic phenotype to serve as a cell-based disease model for study of the disease pathogenesis and evaluation of drug efficacy. The enzyme replacement therapy with recombinant Hex A protein and two small molecules (cyclodextrin and tocopherol) significantly ameliorated lipid accumulation in the Tay-Sachs disease cell model.


Safety surveillance for ustekinumab and other psoriasis treatments from the Psoriasis Longitudinal Assessment and Registry (PSOLAR) Errata.
Erratum for article "Safety surveillance for ustekinumab and other psoriasis treatments from the Psoriasis Longitudinal Assessment and Registry (PSOLAR)." Retrieved from https://jddonline.com/articles/dermatology/S1545961615P0706XJ Drugs Dermatol 2015;14(7):706-714  J Drugs Dermatol 2020;19:e32-e34.


[Antioxidation and multidrug resistance of structure-similar ATP-binding cassette transporter proteins in two fungal entomopathogens].
Structure-similar ATP-binding cassette (ABC) transporter proteins, BbT1 in Beauveria bassiana (Bb) and IfT1 in Isaria fumosorosea (If), were characterized. Various phenotypes including cellular antioxidant response, multidrug resistance and virulence were compared between wild-type strain and the constructed mutants DeltaBbT1, DeltaBbT1/BbT1 and DBbT1/IfT1. Compared with the wild-type, DeltaBbT1/BbT1 and DeltaBbT1/ IfT1 strains showing no significant changes in examined phenotypes, DeltaBbT1 became 27% to 2.1-fold less tolerant to the oxidative stress of 20-40 mmol/L H2O2 and 2-8 mmol/L menadione, 28% to 4.7-fold less resistant to carbendazim, itraconazole, dimetachlone, cycloheximide, ethirimol and 4-nitroquinoline N-oxide, and about 20% less virulent to the second-instar larvae of Spodoptera litura. The ABC transporter proteins BbT1 and IfT1 are similar not only in sequence structure but also in biological function, act as one of the determinants for the multidrug resistance of Bb and If, and also contribute to the fungal virulence due to involvement in cellular antioxidant response.


A morning briefing: setting the stage for a clinically and operationally good day.
This tool can be used in any setting where physicians, nurses, and other disciplines work as a team to manage patients and admission-to-discharge flow.


Cell lineage analyses of epithelia and blood vessels in chimeric mouse embryos by use of an embryonic stem cell line expressing the beta-galactosidase gene.
We have established an embryonic stem (ES) cell line, MS1-EL4, which has the potential to make various tissues in chimeric embryos and, at the same time, expresses the beta-galactosidase gene which was introduced as a good cell marker. To examine cell behavior and lineage during embryogenesis, we injected MS1-EL4 cells into host blastocysts and recovered chimeric embryos at various developmental stages. We examined the distribution of the MS1-EL4 cell derivatives by staining whole embryos with X-gal and by making serial paraffin sections. So far we have obtained the following results: (1) the MS1-EL4 cell line is useful for studying cell lineages because of its ubiquitous expression at least until the mid-gestation stage; (2) cells of the primitive ectoderm and its derivative epithelial tissues continue to intermingle with each other until the late primitive streak stage. Then, at early somite stages, cells of various epithelia stop intermingling and give rise to small coherent clones; (3) blood vessels of the yolk sac are formed by local aggregation of the ancestor cells and those of the embryo proper by proliferation and sprouting from fewer angiogenic cells.


Anterior Lumbar Interbody Fusion Using Reaction Bonded Silicon Nitride Implants: Long-Term Case Series of the First Synthetic Anterior Lumbar Interbody Fusion Spacer Implanted in Humans.
In this study, a historical case series is reported of reaction bonded silicon nitride (Si3N4) implants for anterior lumbar interbody fusion (ALIF) for a patient population of 30 and surgery levels L3/4, L4/5, and/or L5/S1. Before the study, the only work on Si3N4 as a biomedical material was associated preliminary work, which involved animal trials using a rabbit model. The objective was to undertake the first use of Si3N4 as a biomedical material for humans, as an implant for ALIF. The Si3N4 implants were prepared by die-pressing silicon powder and reaction bonding in 95 N2/5 H2 at ∼1400°C for ∼50 hours. The surgeries involved a retroperitoneal approach for L3/4 and L4/5 levels and a transperitoneal approach for L5/S1 level. The patient follow-up involved assessment of radiologic fusion up to 30 years and clinical outcomes to 10 years. The reaction bonded Si3N4 implants were found to be biologically safe and to show high fusion rates with minimal subsidence, no abnormal reaction, and no other complications. The primary outcome measure, visual analog scale back pain, improved from a preoperative mean of 8.4 (range, 6-10) to a mean of 3.7 (range, 0-9) at 5 years and a mean of 4.9 (range, 0-9) at 10 years. The Oswestry Disability Index improved from a preoperative mean of 48 (range, 26-84) to a mean of 35 (range, 4-76) at 10 years. This study confirms that Si3N4 is biologically safe in the long-term, with capacity for excellent radiologic osseointegration.


Drugs on the horizon for diabesity.
The coexistence of type 2 diabetes and obesity presents a complex therapeutic challenge. Future combination tablets may include agents to address diabetes and any accompanying cardiovascular risk factors. Injectable agents that improve glycemic control and facilitate weight loss have recently become available: the soluble amylin analogue pramlintide provides an adjunct to insulin therapy in type 1 and type 2 diabetes, and the incretin mimetic exenatide can enhance prandial insulin release in type 2 diabetes. Orally active inhibitors of the incretin-degrading enzyme dipeptidyl peptidase-IV, agonists of peroxisome proliferator-activated receptor (PPAR)-a and PPAR-g ("dual PPARs"), and the CB1 cannabinoid receptor inhibitor rimonabant are advanced in clinical development. Many novel antidiabetic and antiobesity compounds are emerging in preclinical development.


Validity of a self-administered food frequency questionnaire (FFQ) and its generalizability to the estimation of dietary folate intake in Japan.
In an epidemiological study, it is essential to test the validity of the food frequency questionnaire (FFQ) for its ability to estimate dietary intake. The objectives of our study were to 1) validate a FFQ for estimating folate intake, and to identify the foods that contribute to inter-individual variation of folate intake in the Japanese population. Validity of the FFQ was evaluated using 28-day weighed dietary records (DRs) as gold standard in the two groups independently. In the group for which the FFQ was developed, validity was evaluated by Spearman's correlation coefficients (CCs), and linear regression analysis was used to identify foods with large inter-individual variation. The cumulative mean intake of these foods was compared with total intake estimated by the DR. The external validity of the FFQ and intake from foods on the same list were evaluated in the other group to verify generalizability. Subjects were a subsample from the Japan Public Health Center-based prospective Study who volunteered to participate in the FFQ validation study. CCs for the internal validity of the FFQ were 0.49 for men and 0.29 and women, while CCs for external validity were 0.33 for men and 0.42 for women. CCs for cumulative folate intake from 33 foods selected by regression analysis were also applicable to an external population. Our FFQ was valid for and generalizable to the estimation of folate intake. Foods identified as predictors of inter-individual variation in folate intake were also generalizable in Japanese populations. The FFQ with 138 foods was valid for the estimation of folate intake, while that with 33 foods might be useful for estimating inter-individual variation and ranking of individual folate intake.


Silencing of imprinted CDKN1C gene expression is associated with loss of CpG and histone H3 lysine 9 methylation at DMR-LIT1 in esophageal cancer.
The putative tumor suppressor CDKN1C is an imprinted gene at 11p15.5, a well-known imprinted region often deleted in tumors. The absence of somatic mutations and the frequent diminished expression in tumors would suggest that CDKN1C expression is regulated epigenetically. It has been, however, controversial whether the diminution is caused by imprinting disruption of the CDKN1C/LIT1 domain or by promoter hypermethylation of CDKN1C itself. To clarify this, we investigated the CpG methylation index of the CDKN1C promoter and the differentially methylated region of the LIT1 CpG island (differentially methylated region (DMR)-LIT1), an imprinting control region of the domain, and CDKN1C expression in esophageal cancer cell lines. CDKN1C expression was diminished in 10 of 17 lines and statistically correlated with the loss of methylation at DMR-LIT1 in all but three. However, there was no statistical correlation between CDKN1C promoter MI and CDKN1C expression. Furthermore, loss of CpG methylation was associated with loss of histone H3 lysine 9 (H3K9) methylation at DMR-LIT1. Histone modifications at CDKN1C promoter were not correlated with CDKN1C expression. The data suggested that the diminished CDKN1C expression is associated with the loss of methylation of CpG and H3K9 at DMR-LIT1, not by its own promoter CpG methylation, and is involved in esophageal cancer, implying that DMR-LIT1 epigenetically regulates CDKN1C expression not through histone modifications at CDKN1C promoter, but through that of DMR-LIT1.


Fluorescent characteristics and metal binding properties of individual molecular weight fractions in municipal solid waste leachate.
Molecular weight (MW) is a fundamental property of dissolved organic matter (DOM), which potentially affects the binding behavior between DOM and metals. Here, a combined approach of ultrafiltration fractionation, fluorescence excitation-emission matrix quenching and parallel factor analysis (PARAFAC) was employed to elucidate fluorescent characteristics and metal binding properties of individual MW fractions of DOM in landfill leachate. Four humic-like and two protein-like components were identified by PARAFAC. Among them, a fulvic acid-like component was found to be responsible for Cd(II) binding while Cu(II) inclined to complex with humic-like components rather than protein-like ones. Apart from that, MW was found to exert less influence on metal binding than that of specific metals or components. Key components distributed within various fractions of DOM were the main influence on the impact of MW on metal binding.


24-Epibrassinolide alleviates organic pollutants-retarded root elongation by promoting redox homeostasis and secondary metabolism in Cucumis sativus L.
Environmental pollution by organic pollutants (OPs) has become a global concern due to its detrimental effects on the environment and human health. As plants are used to remediate contaminated sites, understanding the responses of plants to various OPs and fortification of plant tolerance are of great significance. In this work, we studied the biochemical and molecular responses of cucumber plants to three well-known OPs, 2,4,6-trichlorophenol, chlorpyrifos and oxytetracycline in the absence or presence of 24-epibrassinolide (EBR), a potent regulator of plant growth and stress tolerance. The results showed that the selected three OPs retarded root elongation; however, the phytotoxic effects of OPs were attenuated by exogenous EBR. OPs induced accumulations of both hydrogen peroxide (H2O2) and nitric oxide (NO) in root tips and resulted in an increased malondialdehyde (MDA) content, an indicator of membrane lipid peroxidation. Exogenous EBR reduced accumulations of H2O2, NO and MDA in the roots by increasing the expression of antioxidant and detoxification genes and the activities of the corresponding enzymes. Intriguingly, EBR not only promoted the activities of glutathione S-transferase and glutathione reductase, but also increased the content of reduced glutathione without altering the content of oxidized glutathione, which resulted in a reduced redox state under OPs stress. Furthermore, EBR increased the free radical scavenging capacity, flavonoid content and the activity and transcription of secondary metabolism related enzymes. Our results suggest that EBR treatment may fortify secondary metabolism to enhance antioxidant capacity in response to OPs treatment, which might have potential implication in phytoremediation of OPs.


From crystal clear ligand binding to designer I domains.
A long awaited crystal structure of an integrin I domain in complex with a peptide derived from collagen has revealed the ligand-bound conformation of this domain and suggests a mechanism for allosteric control of integrin function by ligand binding. Also, a computational protein design approach has allowed the creation of stable, high affinity forms of the I domain for the first time.


Daily intake of high dietary fiber slows accelerated colonic transit induced by restrain stress in rats.
Stress and central corticotropin releasing factor (CRF) are contributing factors to the pathogenesis of irritable bowel syndrome (IBS). It has been shown that restraint stress and central CRF stimulate colonic motility in rats. Short-chain fatty acids (SCFAs) are produced by bacterial fermentation from dietary fibers. It is controversial whether daily intake of dietary fiber is beneficial for patients with diarrhea-predominant IBS. We studied whether dietary fiber intake affects colonic transit and motility stimulated by restraint stress and central CRF in rats. Corn starch is digested almost completely in the small intestine, while potato starch shows substantial resistance to alpha-amylase and its consumption leads to stimulation of large-bowel fermentation. Four weeks after a corn starch, potato starch, or usual diet (rat chow), colonic transit and motility stimulated by restraint stress and intracisternal (ic) injection of CRF were studied. Fecal concentration of SCFAs in the cecum was measured by high-performance liquid chromatography (HPLC). Four-week treatment with potato starch diet significantly increased cecal concentrations of SCFAs, compared to treatment with corn starch diet. Accelerated colonic transit induced by stress and ic injection of CRF (1 microg) were significantly attenuated in rats receiving potato starch diet compared to rats receiving corn starch diet. The incidence of unformed stool (diarrhea) induced by stress and CRF was also reduced in rats receiving potato starch diet compared to rats receiving corn starch diet and usual diet. It is suggested that daily intake of a high-fiber diet may prevent stress- and CRF-induced acceleration of colonic transit and diarrhea. This study may contribute to treatment for the patients of diarrhea-predominant IBS.


Altered functional connectivity in brain networks underlying self-referential processing in delusions of reference in schizophrenia.
Delusions of reference in schizophrenia are thought to result from misattributions of self-relevance to neutral events. Activation of regions within the cortical midline structures (CMS; e.g., medial prefrontal cortex, cingulate cortex, and precuneus) have been previously associated with self-referential processing in schizophrenia patients; however, the specificity of this pattern to individuals with current delusions of reference has yet to be determined. In the present study, we identified functional brain networks that underlie self-referential processing using task-based multivariate functional connectivity. Healthy control subjects (n=15) and schizophrenia patients with (n=14) and without (n=13) current delusions of reference were shown ambiguous statements while undergoing functional magnetic resonance imaging, and evaluated whether these statements were thought to be specifically about them. Our results revealed two functionally distinct CMS networks that differed between patients and controls during self-referential processing: a posterior CMS network, which showed muted deactivity in non-delusional patients; and an anterior CMS network, in which delusional patients demonstrated hyperactivity. Furthermore, activity within the anterior CMS network across the three groups showed a linear pattern of increasing activity associated with greater intensity of delusions of reference, suggesting that hyperactivity in this network may underlie delusions of reference.


Validation of the Postgraduate Hospital Educational Environment Measure (PHEEM) in a sample of 731 Greek residents.
The Greek version of the Postgraduate Hospital Educational Environment Measure (PHEEM) was evaluated to determine its psychometric properties, i.e., validity, internal consistency, sensitivity and responsiveness to be used for measuring the learning environment in Greek hospitals. The PHEEM was administered to Greek hospital residents. Internal consistency was measured using Cronbach's alpha. Root Mean Square Error of Approximation (RMSEA) was used to evaluate the fit of Structural Equation Models. Content validity was addressed by the original study. Construct validity was tested using confirmatory (to test the set of underlying dimensions suggested by the original study) and exploratory (to explore the dimensions needed to explain the variability of the given answers) factor analysis using Varimax rotation. Convergent validity was calculated by Pearson's correlation coefficient regarding the participant's PHEEM score and participant's overall satisfaction score of the added item "Overall, I am very satisfied with my specialization in this post". Sensitivity was checked by comparing good versus poor aspects of the educational environment and by satisfied versus unsatisfied participants. A total of 731 residents from 83 hospitals and 41 prefectures responded to the PHEEM. The original three-factor model didn't fit better compared to one factor model that is accounting for 32% of the variance. Cronbach's α was 0.933 when assuming one-factor model. Using a three-factor model (autonomy, teaching, social support), Cronbach's α were 0.815 (expected 0.830), 0.908 (0.839), 0.734 (0.793), respectively. The three-factor model gave an RMSEA value of 0.074 (90% confidence interval 0.071, 0.076), suggesting a fair fit. Pearson's correlation coefficient between total PHEEM and global satisfaction was 0.765. Mean question scores ranged from 19.0 (very poor) to 73.7 (very good), and mean participant scores from 5.5 (very unsatisfied) to 96.5 (very satisfied). The Greek version of PHEEM is a valid, reliable, and sensitive instrument measuring the educational environment among junior doctors in Greek hospitals and it can be used for evidence-based SWOT analysis and policy.


Bowen's disease of the nail apparatus: a series of 8 patients and a literature review.
Bowen's disease of the nail apparatus is uncommon. Since often characteristic clinical features are missing, diagnosis and treatment are delayed. We have analyzed our patients' files from January 2001 to July 2015 for this disorder. We identified eight patients with Bowen's disease of the nail apparatus, six male and two female individuals. Fingers were more often affected than toes. The disease does not respect the anatomical borders of the nail apparatus. Therefore, we performed delayed Mohs surgery with skin grafts in seven patients and with second intention healing in one patient. Two relapses were noted but only one ate the same digit. Since relapses occurred after 2-3 years, a follow-up of such patients seems justified.


Kidney tubular transcription co-activator, Yes-associated protein 1 (YAP), controls the expression of collecting duct aquaporins and water homeostasis.
Final urine volume and concentration are defined by water reabsorption through the water channel proteins aquaporin (AQP)-2, -3 and -4 in the collecting duct. However, the transcriptional regulation of these AQPs is not well understood. The Hippo/Yes-associated protein 1 (YAP) pathway plays an important role in organ size control and tissue homeostasis. When the Hippo pathway including the Mst1/Mst2 kinases is inhibited, YAP is activated and functions as a transcription co-activator. Our previous work revealed a pathological role of tubular YAP activation in chronic kidney disease, but the physiological role of YAP in the kidney remains to be established. Here, we found that tubule-specific Yap knockout mice showed increased urine output and decreased urinary osmolality. Decreases in Aqp2, -3 and -4 mRNA and protein abundance in the kidney were evident in Yap knockout mice. Analysis of Mst1/Mst2 double knockout and Mst1/Mst2/Yap triple knockout mice showed that expression of Aqp2 and Aqp4 but not Aqp3 was dependent on YAP. Furthermore, YAP was recruited to the promoters of the Aqp2 and Aqp4 genes and stimulated their transcription. Interestingly, YAP was found to interact with transcription factors GATA2, GATA3 and NFATc1. These three factors promoted Aqp2 transcription in a YAP dependent manner in collecting duct cells. These three factors also promoted Aqp4 transcription whereas only GATA2 and GATA3 enhanced Aqp3 transcription. Thus, our results suggest that YAP promotes Aqp2 and Aqp4 transcription, interacts with GATA2, GATA3 and NFATc1 to control Aqp2 expression, while Aqp-2, -3 and -4 exploit overlapping mechanisms for their baseline transcriptional regulation.


Daikenchuto accelerates the recovery from prolonged postoperative ileus after open abdominal surgery: a subgroup analysis of three randomized controlled trials.
Prolonged postoperative ileus (POI) is a common complication after open abdominal surgery (OAS). Daikenchuto (DKT), a traditional Japanese medicine that peripherally stimulates the neurogenic pathway, is used to treat prolonged POI in Japan. To analyze whether DKT accelerates the recovery from prolonged POI after OAS, we conducted a secondary analysis of three multicenter randomized controlled trials (RCTs). A secondary analysis of the three RCTs supported by the Japanese Foundation for Multidisciplinary Treatment of Cancer (project numbers 39-0902, 40-1001, 42-1002) assessing the effect of DKT on prolonged POI in patients who had undergone OAS for colon, liver, or gastric cancer was performed. The subgroup included 410 patients with no bowel movement (BM) before the first diet, a DKT group (n = 214), and a placebo group (n = 196). Patients received either 5 g DKT or a placebo orally, three times a day. The primary endpoint was defined as the time from the end of surgery to the first bowel movement (FBM). A sensitivity analysis was also performed on the age, body mass index and dosage as subgroup analyses. The primary endpoint was significantly accelerated in the DKT group compared with the placebo group (p = 0.004; hazard ratio 1.337). The median time to the FBM was 113.8 h in the placebo group and 99.1 h in the DKT treatment group. The subgroup analysis showed that DKT significantly accelerated the recovery from prolonged POI following OAS. UMIN000026292.


Real-time structure search and structure classification for AlphaFold protein models.
Last year saw a breakthrough in protein structure prediction, where the AlphaFold2 method showed a substantial improvement in the modeling accuracy. Following the software release of AlphaFold2, predicted structures by AlphaFold2 for proteins in 21 species were made publicly available via the AlphaFold Database. Here, to facilitate structural analysis and application of AlphaFold2 models, we provide the infrastructure, 3D-AF-Surfer, which allows real-time structure-based search for the AlphaFold2 models. In 3D-AF-Surfer, structures are represented with 3D Zernike descriptors (3DZD), which is a rotationally invariant, mathematical representation of 3D shapes. We developed a neural network that takes 3DZDs of proteins as input and retrieves proteins of the same fold more accurately than direct comparison of 3DZDs. Using 3D-AF-Surfer, we report structure classifications of AlphaFold2 models and discuss the correlation between confidence levels of AlphaFold2 models and intrinsic disordered regions.


Molecular cloning of liver angiotensinogen gene in ayu (Plecoglossus altivelis) and mRNA expression changes upon Aeromonas hydrophila infection.
Angiotensinogen (AGT) is thought to be an acute phase protein, because in mammals its plasma concentrations increase during inflammation induced by lipopolysaccharides (LPS). A full-length cDNA clone of the AGT gene, 1828bp in size, was isolated from the fish ayu (Plecoglossus altivelis). Its deduced amino acid sequence of 462 residues had 57.7-68.6%, 30.2% and 23.3-27.4% identity to those sequenced from other fish, chicken and mammals, respectively. Ayu-AGT is most closely related to that of silver sea bream and the phylogenetic tree largely reflects the currently accepted animal relationships. In healthy control fish, AGT transcripts were found to be present in a number of tissues including brain, spleen, liver, kidney and intestine. The amplification was found to be the highest in the liver. In Aeromonas hydrophila-infected ayu, AGT transcripts significantly increased in various tissues, suggesting a possible relationship between AGT mRNA expression and the vascular inflammation caused by bacterial infection. AGT transcripts could be detected in the heart and gill of bacteria-infected ayu that were not detected in control fish, but still not in the muscle. This is the first report of changes in fish AGT transcripts upon bacterial infection.


Primary structure of a pheromone-binding protein from Antheraea pernyi: homologies with other ligand-carrying proteins.
An antennal cDNA clone encoding the complete sequence (163 amino acids) of a pheromone-binding protein precursor from the male silk moth, Antheraea pernyi, was isolated using oligonucleotide probes. The cloned cDNA was expressed and the translation product detected by specific antibodies. The deduced protein sequence consists of a signal peptide of 21 amino acids and a mature binding protein of 142 amino acid residues. The predicted structure of this protein is homologous to binding-proteins from different insect species which have previously been identified, but shows no similarities to odorant-binding proteins from vertebrates, suggesting that soluble odorant-binding proteins in insects and vertebrates represent an evolutionary convergence.


Journal submissions, review and editorial decision patterns during initial COVID-19 restrictions.
We use the full administrative records from four leading agricultural economics journals to study the impacts of the COVID-19 pandemic on manuscript submission, editorial desk rejection and reviewer acceptance rates, and time to editorial decision. We also test for gender differences in these impacts. Manuscript submissions increased sharply and equi-proportionately by gender. Desk rejection rates remained stable, leading to increased demand for reviews. Female reviewers became eight percentage points more likely to decline a review invitation during the early stage of the pandemic. First editorial decisions for papers sent out for peer review occurred significantly faster after pandemic lockdowns began. Overall, the initial effects of the pandemic on journal editorial tasks and review patterns appear relatively modest, despite the increased number of submissions handled by editors and reviewers. We find no evidence in agricultural economics of a generalized disruption to near-term, peer-reviewed publication.


Incidence of histologic chorioamnionitis in a rural community hospital.
Histological chorioamnionitis (HCA) is a pathological condition defined as an acute inflammation of the amniochorionic membranes which has been linked to a wide range of adverse neonatal events. The purpose of this study is to identify the incidence of HCA in the rural population and evaluate whether there are significant differences in these incidences within the different clinical parameters of delivery method, gravidity, gestational age, previous cesarean section, reason for cesarean section and body mass index (BMI). A retrospective chart review was conducted on 462 consecutive deliveries that occurred in a rural hospital during a four-year period. Data collected was analyzed using independent sample T-tests, chi-squared tests, and descriptive statistics, with a p-value of < 0.05. Overall incidence of HCA was 15.9% (73/459), with a term incidence of 16.2% (68/421) and preterm incidence of 13.2% (5/38). The incidence of HCA was significantly higher in vaginal deliveries (18.8%; n = 54/288) than C-section deliveries (11.1%; n = 19/171) (p = 0.03). Incidence of clinical chorioamnionitis was 0.43% (2/462), with 2.74% (2/73) of HCA manifesting clinically. Evaluation of the incidence of HCA and associated clinical parameters in this study showed a marked decrease in the incidence of HCA when compared to other studies. Strategies to reduce the incidence of HCA include reducing the length of labor via active labor management. We hypothesize that these findings are due to the consistent use of active labor management and our rural study population, but further investigation is required to confirm this.


First genetic detection and characterization of canine parvovirus type 2 (Carnivore protoparvovirus 1) in southwestern Ethiopia.
The species Carnivore protoparvovirus 1 includes viruses, e.g. canine parvovirus (CPV-2) and feline panleukopenia virus (FPV), which are among the most relevant for pets, causing extremely severe clinical signs and high fatality rate in dogs and cats. Moreover, a broad range of wild hosts, including endangered ones, were proven to be susceptible. Currently, no data on CPV-2 molecular epidemiology and strain characterization are available in Ethiopia, also considering the frequent contacts between domestic and wild populations. In the present study, a molecular epidemiology survey was performed on 92 fecal samples collected from domestic (n = 84) and stray (n = 8) dogs in southwestern Ethiopia in 2021. Approximately, 10% of the samples tested positive and the complete VP2 sequences of 5 strains were obtained, classified within the CPV-2a (n = 1) and CPV-2c (n = 4) antigenic variants. In most instances, the closest genetic relatives were strains of Chinese origin, which is fully compatible with the intense relationships that have been developing between the two countries, involving human being travels and their pets as well. Considering the clinical relevance of this infection and the risk it poses to local domestic and wild carnivore populations, especially endangered ones, much stronger monitoring and surveillance activity on foreign incoming animals should be performed. More strict constraints on animal introduction, e.g. compulsory vaccination, should also be considered.


Microvascular decompression treatment for post-Bell's palsy hemifacial spasm.
This retrospective study is to explore the clinical features and surgical outcomes of the patients who suffered from hemifacial spasm preceded by Bell's palsy. Seventeen patients with post-Bell's palsy hemifacial spasm underwent microvascular decompression surgery. A 3D-TOF-MRA examination was performed pre-operatively to confirm the existence of offending vessels around the facial nerve. Abnormal muscle response was monitored during operation. The results of spasm resolution and post-operative complications were assessed. During operation, offending vessels were found and transposed in 15 patients. For the other two patients in whom offending vessel was absent, the facial nerve was treated by combing. The results of spasm resolution were 'cured' in 12 (70·5%) patients, 'improved' in 2 (11·8%) patients, 'fair' in 2 (11·8%) patient, and 'failed' in 1 (5·9%) patient. The complications included transient hearing loss in one case, and deterioration of facial weakness in two cases. Vascular compression may be an etiological factor of post-Bell's palsy hemifacial spasm, and microvascular decompression is an effective treatment to this disorder.


Complications of modeling glycosylation reactions: can the anomeric conformation of a donor determine the glycopyranosyl oxacarbenium ring conformation?
That the ring conformation of glycopyranosyl oxacarbenium ions can influence the stereochemical outcome of glycosylation reactions has been postulated for some time. Some new ionization calculations show that the ultimate conformation (4)H(3) or (5)S(1) of D-glucopyranosyl oxacarbenium ions depends on the initial ϕ(H) (CH-1-C-1-S(+)-SCH(3)) conformation of anomeric thiosulfonium ions. Evidence is also presented that nucleophile:electrophile hydrogen bonded complexes, 1,6-anhydro-carbenium ions and electron rich carbon nucleophile:oxacarbenium ion complexes are all probably artifacts of neglecting counter ions or nucleophiles in the DFT calculation. All three cationic species are likely important for glycosylation reaction side reactions but not as productive species.


Evolving a strategic approach to cervical cancer control in Africa.
Nigeria is a country in Western Africa where the incidence rate of cervical cancer is 25/100,000. There are 32 million women aged 15-64 years old. If we were to conduct a one-time screen over 1 year, 8000 new invasive cervical cancers would be detected. Currently, 80% of cases present in Stage III. While a one-time screen should detect some earlier stages, there could be as many as 6400 Stage III cancers to treat. Strategies to enable a one-time screen are being considered.


[Prospective evaluation of PICC line related complications].
To report the occurrence of complications following placement of peripherally inserted central catheters (PICC). Materials and methods. A total of 127 PICC lines were placed in 115 patients for TPN (n=54), long-term antibiotics (n=29), chemotherapy (n=14) or miscellaneous intravenous therapy (n=18). All PICC lines were successfully inserted. PICC lines were used for a mean duration of 16 days (1-166 days). The following complications were recorded: occlusion (7%), rupture (1.6%), accidental withdrawal (2.4%), infection (3.1%) and venous thrombosis (2.4%). PICC lines are a simple, safe and effective alternative to conventional central venous catheters. The occurrence of complications, typically "mechanical", must be reduced and prevented by strict management of this type of central line by the treating team.


Long noncoding RNA CCHE1 promotes cervical cancer cell proliferation via upregulating PCNA.
Long noncoding RNAs (lncRNAs) have been shown to play important roles in carcinogenesis and progression. However, the roles and functional mechanisms of lncRNAs in cervical cancer remain largely unknown. In this study, we found that cervical carcinoma high-expressed lncRNA 1 (lncRNA-CCHE1) was significantly upregulated in cervical cancer tissues. The higher expression of CCHE1 was significantly correlated with large tumor size, advanced Federation of Gynecology and Obstetrics stage, uterine corpus invasion, and poor survival. Gain-of-function and loss-of-function experiments demonstrated that CCHE1 overexpression promotes the proliferation of cervical cancer cell. By contrast, the depletion of CCHE1 inhibits the proliferation of cervical cancer cells. RNA pull-down assays confirmed that CCHE1 physically associates with proliferating cell nuclear antigen (PCNA) messenger RNA, consequently enhances the expression of PCNA. The expression of CCHE1 and PCNA is significantly correlated in cervical cancer tissues. The depletion of PCNA abolishes the effects of CCHE1 on the proliferation of cervical cancer cells. Taken together, these findings indicate that CCHE1 plays a pivotal role in cervical cancer cell proliferation via increasing PCNA expression and serves as a potential prognostic biomarker and therapeutic target in human cervical cancer.


A new species of Lycosa and three new species and a new record of Karakumosa from Iran (Araneae, Lycosidae).
Four new species of lycosids from Iran are described: Lycosa soboutii sp. n. (, Zanjan Province), Karakumosa golestanica sp. n. (, Golestan Province), K. sarvari sp. n. (, Razavi Khorasan Province), and K. yahaghii sp. n. (, South Khorasan Province). Karakumosa shmatkoi Logunov Ponomarev, 2020 (, Razavi Khorasan Province) is reported from Iran for the first time, and its male is redescribed.


Topographical anatomy of the albino rat's ischiotrochanteric muscle group.
The obturator internus, gemelli, and quadratus femoris muscles are thought to be postural muscles. Thus, they are in the focus of research. Although these muscles are described in other species, detailed descriptions are still lacking for the albino rat. We hypothesized that the rat's ischiotrochanteric muscle group is comparable to that of humans. We aimed to provide a detailed description, and to compare the rat's condition with other species including humans. Both hind limbs of 30 adult male formalin-fixed albino rats were carefully dissected and photo documented. Our results were then compared with data for other species and descriptions of human anatomy. The gemellus muscle was one single muscle mass, originating from the lesser sciatic notch and an unnamed groove on the dorsal aspect of the ischium. The obturator internus muscle arose from the inner aspect of the tabula of ischium. Both muscles formed a continuum and inserted as one complex on the medial aspect of the greater trochanter. The quadratus femoris muscle originated from the outer aspect of the tabula of ischium and inserted on the distal portion of the intertrochanteric crest, and the dorsal aspect of the lesser trochanter. Despite minor differences, the topographical conditions of these muscles are comparable between rats and other mammals including humans.


Multi-omics strategies uncover the molecular mechanisms of nitrogen, phosphorus and potassium deficiency responses in Brassica napus.
Nitrogen (N), phosphorus (P) and potassium (K) are critical macronutrients in crops, such that deficiency in any of N, P or K has substantial effects on crop growth. However, the specific commonalities of plant responses to different macronutrient deficiencies remain largely unknown. Here, we assessed the phenotypic and physiological performances along with whole transcriptome and metabolomic profiles of rapeseed seedlings exposed to N, P and K deficiency stresses. Quantities of reactive oxygen species were significantly increased by all macronutrient deficiencies. N and K deficiencies resulted in more severe root development responses than P deficiency, as well as greater chlorophyll content reduction in leaves (associated with disrupted chloroplast structure). Transcriptome and metabolome analyses validated the macronutrient-specific responses, with more pronounced effects of N and P deficiencies on mRNAs, microRNAs (miRNAs), circular RNAs (circRNAs) and metabolites relative to K deficiency. Tissue-specific responses also occurred, with greater effects of macronutrient deficiencies on roots compared with shoots. We further uncovered a set of common responders with simultaneous roles in all three macronutrient deficiencies, including 112 mRNAs and 10 miRNAs involved in hormonal signaling, ion transport and oxidative stress in the root, and 33 mRNAs and 6 miRNAs with roles in abiotic stress response and photosynthesis in the shoot. 27 and seven common miRNA-mRNA pairs with role in miRNA-mediated regulation of oxidoreduction processes and ion transmembrane transport were identified in all three macronutrient deficiencies. No circRNA was responsive to three macronutrient deficiency stresses, but two common circRNAs were identified for two macronutrient deficiencies. Combined analysis of circRNAs, miRNAs and mRNAs suggested that two circRNAs act as decoys for miR156 and participate in oxidoreduction processes and transmembrane transport in both N- and P-deprived roots. Simultaneously, dramatic alterations of metabolites also occurred. Associations of RNAs with metabolites were observed, and suggested potential positive regulatory roles for tricarboxylic acids, azoles, carbohydrates, sterols and auxins, and negative regulatory roles for aromatic and aspartate amino acids, glucosamine-containing compounds, cinnamic acid, and nicotianamine in plant adaptation to macronutrient deficiency. Our findings revealed strategies to rescue rapeseed from macronutrient deficiency stress, including reducing the expression of non-essential genes and activating or enhancing the expression of anti-stress genes, aided by plant hormones, ion transporters and stress responders. The common responders to different macronutrient deficiencies identified could be targeted to enhance nutrient use efficiency in rapeseed.


Genome shuffling leads to rapid phenotypic improvement in bacteria.
For millennia, selective breeding, on the basis of biparental mating, has led to the successful improvement of plants and animals to meet societal needs. At a molecular level, DNA shuffling mimics, yet accelerates, evolutionary processes, and allows the breeding and improvement of individual genes and subgenomic DNA fragments. We describe here whole-genome shuffling; a process that combines the advantage of multi-parental crossing allowed by DNA shuffling with the recombination of entire genomes normally associated with conventional breeding. We show that recursive genomic recombination within a population of bacteria can efficiently generate combinatorial libraries of new strains. When applied to a population of phenotypically selected bacteria, many of these new strains show marked improvements in the selected phenotype. We demonstrate the use of this approach through the rapid improvement of tylosin production from Streptomyces fradiae. This approach has the potential to facilitate cell and metabolic engineering and provide a non-recombinant alternative to the rapid production of improved organisms.


Carbaxylosides of 4-ethyl-2-oxo-2H-benzopyran-7-yl as non-hydrolyzable, orally active venous antithrombotic agents.
A (-)-conduritol F derivative was condensed with 4-ethyl-7-hydroxy-2H-1-benzopyran-2-one and converted into (+)-4-ethyl-7-[(1'R,2'S,3'S,4'R)-2',3',4'- trihydroxycyclohexyloxy]-2H-1-benzopyran-2-one ((+)-2). Enantiomer (-)-2 was obtained from a (+)-conduritol F derivative. The carbaxyloside (-)-2 with the L-xylose configuration was more active than (+)-2 in the Wessler's model.


ApoE4 impairs hippocampal plasticity isoform-specifically and blocks the environmental stimulation of synaptogenesis and memory.
Alzheimer's disease (AD) is associated with genetic risk factors, of which the allele E4 of apolipoprotein E (apoE4) is the most prevalent, and is affected by environmental factors that include education early in life and socioeconomic background. The extent to which environmental factors affect the phenotypic expression of the AD genetic risk factors is not known. Here we show that the neuronal and cognitive stimulations, which are elicited by environmental enrichment at a young age, are markedly affected by the apoE genotype. Accordingly, exposure to an enriched environment of young mice transgenic for human apoE3, which is the benign AD apoE allele, resulted in improved learning and memory, whereas mice transgenic for human apoE4 were unaffected by the enriched environment and their learning and memory were similar to those of the nonenriched apoE3 transgenic mice. These cognitive effects were associated with higher hippocampal levels of the presynaptic protein synaptophysin and of NGF in apoE3 but not apoE4 transgenic mice. In contrast, cortical synaptophysin and NGF levels of the apoE3 and apoE4 transgenic mice were similarly elevated by environmental enrichment. These findings show that apoE4 impairs hippocampal plasticity and isoform-specifically blocks the environmental stimulation of synaptogenesis and memory. This provides a novel mechanism by which environmental factors can modulate the function and phenotypic expression of the apoE genotype.


Factors associated with human papillomavirus infection among women in the Northwest Territories.
In some regions, Aboriginal women have higher rates of cervical cancer compared to other Canadian women. However, little data are available regarding the co-factors of HPV infection in the Aboriginal population of Canada. We examined factors associated with high-risk human papillomavirus (HR-HPV) infection in a population of women (both Aboriginal and non-Aboriginal) in the Northwest Territories (NWT). We used a cross-sectional design using a convenience sample of all women aged 14 years or older presenting for Papanicolaou (Pap) testing across all regions of the NWT from March 2009 to March 2010. Women answered a questionnaire on demographic characteristics, sexual behaviour, and gynaecological and obstetrical events. We used multiple regression analysis to explore factors associated with HR-HPV infection according to age and cultural background. HPV typing was done using the Luminex assay. Of the total 1,279 participants, 178 had missing HPV results. We obtained data on 1,101 women and overall HR-HPV prevalence was 14.2%. Younger age, single marital status, Aboriginal background, current smoking, lifetime deliveries, use of hormonal contraceptives, and the numbers of sexual partners in the last year were associated with prevalent HR-HPV. Our findings tend to indicate that Aboriginal women have different predictors of HR-HPV than non-Aboriginal women that may affect HPV progression to cervical cancer. These findings can help to better target public health practices for the women at higher risk of HPV infection and cervical cancer.


[Profiles of urethral pressure, measured with microtransducers, in the assessment of female stress urinary incontinence].
This study was conducted to evaluate the usefulness of urethral pressure profiles in the diagnosis of urinary stress incontinence and to determine the possible correlation of such profiles with the degree of incontinence. An initial group of 92 female patients were studied; 29 that presented vesical instability in the urodynamic studies were posteriorly discarded from the study. Patient evaluation included clinical history, physical examination, analytical, radiological and urodynamic studies, including uroflowmetry, filling and emptying cystometry and static and dynamic urethral pressure profiles. A Phoenix 5 and a 10 F microtransducter catheter were used to carry out the necessary evaluations. ICS recommendations were observed. In the statistical analysis of the results, the Wilconson test and contingency table of 2x2 were employed. In the different groups, the parameters analyzed in the static urethral pressure profiles revealed only statistically significant differences in maximum pressure urethral values and maximum closing urethral pressure. No differences in either total length or in functional urethral length were observed. The values of the dynamic urethral pressure profiles of the different groups were compared. There was a statistically significantly higher proportion of patients that presented a negative closing urethral pressure dynamics in the group of women with urodynamically and clinically demonstrated urinary incontinence in comparison to the group of women whose incontinence was not demonstrated either by urodynamic or physical explorations. The urethral pressure profile is sufficiently reliable in confirming both the diagnosis of urinary incontinence and its degree of severity. As a diagnostic test for urinary stress incontinence, it has a sensitivity of 82% and a specificity of 90%.


Neuroimaging.
Imaging is integral to the management of patients with brain tumors. Conventional structural imaging provides exquisite anatomic detail but remains limited in the evaluation of molecular characteristics of intracranial neoplasms. Quantitative and physiologic biomarkers derived from advanced imaging techniques have been increasingly utilized as problem-solving tools to identify glioma grade and assess response to therapy. This chapter provides a comprehensive overview of the imaging strategies used in the clinical assessment of patients with gliomas and describes how novel imaging biomarkers have the potential to improve patient management.


Substance P activates responses correlated with tumour growth in human glioma cell lines bearing tachykinin NK1 receptors.
The neuropeptide substance P (SP), by stimulating tachykinin NK1 receptors (NK1R), triggers a number of biological responses in human glioma cells which are potentially relevant for tumour growth. First, radioligand binding studies demonstrated the presence of tachykinin NK1R on SNB-19, DBTRG-05 MG and U373 MG, but not on U138 MG and MOG-G-GCM human glioma cell lines. Second, application of SP or neurokinin A (NKA) to NK1R+ glioma cell lines increased the secretion of interleukin 6 (IL-6) and potentiated IL-6 secretion induced by IL-1beta. SP also up-regulated the release of transforming growth factor beta1 (TGF-beta1) by the U373 MG glioma cell line. Third, SP induced new DNA synthesis and enhanced the proliferation rate of NK1R+, but not of NK1R- glioma cell lines. Also, NKA stimulated the proliferation and cytokine secretion in NK1R+ glioma cell lines. All the stimulant effects of SP/NKA on NK1R+ glioma cell lines were completely blocked by a specific tachykinin NK1R antagonist, MEN 11467. These data support the potential use of tachykinin NK1R antagonist for controlling the proliferative rate of human gliomas.


[Value of blood lipase determination in pancreatic diseases. Comparison with blood amylase assay].
The comparative diagnostic value of plasma lipase and amylase levels was evaluated in 90 patients with various symptoms of pancreatic disease. Comparison with healthy subjects showed that the lipase assay was more sensitive than the amylase assay in patients with acute pancreatitis or post-traumatic pancreatic injury, but not in patients with chronic pancreatitis. On the other hand, acute pancreatitis can be detected earlier by measuring plasma amylase levels, owing to the rapid release of this enzyme. The relative importance of these two laboratory tests is discussed.


Purification and immunological analysis of RNA polymerase II from Caenorhabditis elegans.
We describe a rapid procedure for obtaining highly purified RNA polymerase II from the nematode Caenorhabditis elegans. The structure of the enzyme was examined by denaturing gel electrophoresis and found to consist of three large polypeptides (molecular weights 200,000, 175,000, and 135,000) and eight smaller polypeptides (molecular weights 29,500, 20,000, 16,000, 15,000, 13,000, 11,500, 10,500, and 9,500). As observed for the analogous enzyme from other organisms, the 175,000 polypeptide (II175) appeared to be a degraded form of the 200,000 polypeptide (II200). The structure of nematode RNA polymerase II closely resembles that of the corresponding enzyme from other animals. Four of its larger subunits shared antigenicity with Drosophila RNA polymerase II. Antibody raised against purified RNA polymerase II reacted with several enzyme subunits in "Western" blots of purified polymerase and impure enzyme fractions. Immunofluorescence staining was used to visualize RNA polymerase II in the nuclei of a nematode squash preparation and the nucleoplasm of cultured mammalian cells.