Pushing antibody-based labeling systems to higher sensitivity by linker-assisted affinity enhancement.
The sensitivity of antibody/hapten-based labeling systems is limited by the natural affinity ceiling of immunoglobulins. Breaking this limit by antibody engineering is difficult. We thus attempted a different approach and investigated if the so-called bridge effect, a corecognition of the linker present between hapten and carrier protein during antibody generation, can be utilized to improve the affinity of such labeling systems. The well-known haptens 2,4-dinitrophenol (2,4-DNP) and 2,4-dichlorophenoxyacetic acid (2,4-D) were equipped with various linkers, and the resulting affinity change of their cognate antibodies was analyzed by ELISA. Anti-2,4-DNP antibodies exhibited the best affinity to their hapten when it was combined with aminobutanoic acid or aminohexanoic acid. The affinity of anti-2,4-D antibodies could be enhanced even further with longer aliphatic spacers connected to the hapten. The affinity toward aminoundecanoic acid-2,4-D derivatives, for instance, was improved about 100-fold compared to 2,4-D alone and yielded detection limits as low as 100 amoles of analyte. As the effect occurred for all antibodies and haptens tested, it may be sensible to implement the bridge effect in future antibody/hapten-labeling systems in order to achieve the highest sensitivity possible.


Characterization of DNA Condensation by Conformationally Restricted Dipeptides and Gene Delivery.
A wide variety of non-viral vectors have been developed for gene delivery in past few decades but find limited applications mainly due to lower encapsulation, endosomal entrapment, high toxicity and low transfection efficiency. In this work, we explored plasmid DNA binding ability of several low molecular weight dipeptides containing α,β-dehydrophenylalanine (ΔF) and found that an arginine containing dipeptide, arginine-α,β-dehydrophenylalanine (R-ΔF) condensed pEGFP-N1 plasmid into positively charged spherical nanoparticles of size 250–275 nm. Single molecule techniques showed that R-ΔF interacted with the plasmid DNA in a dose dependent manner which was accompanied by a decrease in diffusion time of the plasmid DNA as well as release of the bound fluorophore. The plasmid DNA in R-ΔF-plasmid complex (R-ΔF-Pl) was stable against DNase action. A pH dependent release of the plasmid DNA from R-ΔF-Pl was observed and the released plasmid DNA retained its natural conformation at endosomal pH, as evidenced from time correlated single photon counting. R-ΔF-Pl was biocompatible and showed ready uptake in HEK 293T cells. Transfection assays using reporter plasmids for green fluorescent protein (GFP), luciferase enzyme and chloramphenicol acetyltransferase (CAT) showed R-ΔF mediated gene delivery both in the presence and absence of serum in the medium. Ease of synthesis, homogenous assembly and biocompatibility balanced with significant expression of gene of interest make R-ΔF a potential vector for development for in vivo application.


Metabolism of putrescine in neuroblastoma and glioma cells during culture.
The metabolism of putrescine in neuroblastoma and glioma cells was analyzed during culture. In the former cells the formation of gamma-aminobutyric acid from putrescine was low and constant during the logarithmic phase and increased dramatically during the stationary phase or in cultures with low serum concentrations, while in the latter cells it was low and constant throughout culture. The formation of spermidine from putrescine in both cell lines was active at days 1-2 of culture and decreased gradually during culture. These findings indicate that spermidine formation is closely related to proliferation of the cells, and gamma-aminobutyric acid formation to differentiation or maturation of the neuroblastoma cells.


Epilepsy, Psychogenic Seizure, Trauma in the Developmental Process.
An epileptic seizure, can cause trauma for its sudden emergence, leading to functional impairment, accidents and injuries, and fear of death. The seizure can be traumatizing itself, besides, an head trauma that may occur during the seizure can also cause epilepsy. As the severity and duration of epilepsy increases, disturbances in development and traumatic effects occur. Conversion (psychogenic) seizures may be added over the years in epileptic patients. The comorbidity of trauma-related dissociative disorder and psychogenic seizures is observed in approximately half of the cases. Dissociative disorders are known to occur in children with chronic diseases due to the traumatic effect of the disease. Conversion disorder and psychogenic seizures are frequently seen in dissociative disorders. Posttraumatic stress disorder, dissociative disorders, and psychogenic seizures are often comorbid diagnoses in epilepsy. For this reason, traumatic effect and associated dissociative disorder dimension should be kept in mind in the psychiatric approach when handling with cases of epilepsy.


Comparison of a respiratory suspension aerosolized by an air-jet and an ultrasonic nebulizer.
In the absence of USP standards and performance monographs, this research sought to determine if differences in the aerosolization mechanism (air-jet vs. ultrasonic) affected droplet and insoluble particle deposition of a nebulized model respiratory suspension. Five milliliters of a model suspension containing 0.1% w/v fluorescein (to estimate droplet deposition) and known quantities of 1, 3, and 6 microns latex spheres (representing insoluble drug particles) was aerosolized from an air-jet and an ultrasonic nebulizer. Nebulized output was collected in a modified Andersen impactor. Samples were analyzed spectrophotometrically (490.5 nm) and by a Coulter Counter to estimate droplet and sphere deposition, respectively. The distribution of droplets throughout the modified impactor for both nebulizers suggested that both the air-jet and the ultrasonic nebulizer produced droplets (0.4 to 10 microns in aerodynamic diameter) large enough to incorporate 1, 3, and 6 microns insoluble spheres. However, Coulter Counter analysis of the sphere distribution revealed that while the air-jet nebulized output contained spheres of all sizes, this was not true for the ultrasonic nebulizer. In the ultrasonic nebulizer, 99% of the spheres (irrespective of size) were not aerosolized and were recovered from the nebulizer reservoir at the aerosolization end point. The results highlight the importance of evaluating performance of a respiratory suspension in combination with a specific nebulizer. When conducting in vitro inertial deposition testing of a respiratory suspension, it is inappropriate to assume that deposition trends of droplets will predict the deposition of the insoluble dispersed phase.


Doppler criteria for intrauterine growth retardation: predictive values.
Published data concerning proposed antenatal Doppler criteria for intrauterine growth retardation (IUGR) were critically analyzed using the same technique that has been previously applied to conventional (non-Doppler) sonographic criteria for IUGR. Fifteen studies of Doppler criteria, each conducted in such a way that sensitivity and specificity could be determined, were identified through a literature review. These studies encompassed a variety of arterial waveform and volume blood flow criteria. Bayes' theorem was then used to determine the positive predictive value of each criterion if used as a screening test for IUGR. Criteria involving vessels that have been studied by at least two groups, including those involving umbilical artery waveform, uterine arcuate artery waveform, and umbilical vein volume blood flow, had positive predictive values that fell in a low range (17-57%) similar to that previously found for conventional criteria. Two criteria, using fetal internal carotid artery and thoracic aortic waveforms, had higher positive predictive values (66 and 100%, respectively) in initial studies, but the data have yet to be replicated by other groups. It is concluded that no Doppler criterion has yet been established as a clinically useful method to screen for IUGR antenatally.


Comparison of the changes in central catecholamine systems following short- and long-term lithium treatment and the consequences of lithium withdrawal.
The effects of treatment with and withdrawal from lithium (Li, 2 mmol/kg) on central catecholamine systems in rat brain were investigated. Synthetic parameters were found to be unchanged after treatment, but abrupt withdrawal resulted in enhanced activity of tyrosine hydroxylase. Noradrenaline levels in most brain regions examined were depressed following both short- and long-term Li administration, with a further decrease in content in the pons but enhanced cortical levels on withdrawal. Li administration resulted in elevated 3,4-dihydroxyphenylacetic acid levels, whereas withdrawal caused an enhancement of homovanillic acid content and depression of 3-methoxy-4-hydroxyphenyl glycol levels. It appears that the changes in central catecholamine systems are qualitatively the same following both short- and long-term Li treatment, as are the consequences of abrupt Li withdrawal. It is suggested that Li administration results in a disruption of catecholamine storage processes, while withdrawal from Li does not result in a simple return towards normal states.


Dendrimers Show Promise for siRNA and microRNA Therapeutics.
The lack of an appropriate intracellular delivery system for therapeutic nucleic acids (TNAs) is a major problem in molecular biology, biotechnology, and medicine. A relatively new class of highly symmetrical hyperbranched polymers, called dendrimers, shows promise for transporting small TNAs into both cells and target tissues. Dendrimers have intrinsic advantages for this purpose: their physico-chemical and biological properties can be controlled during synthesis, and they are able to transport large numbers of TNA molecules that can specifically suppress the expression of single or multiple targeted genes. Numerous chemical modifications of dendrimers extend the biocompatibility of synthetic materials and allow targeted vectors to be designed for particular therapeutic purposes. This review summarizes the latest experimental data and trends in the medical application of various types of dendrimers and dendrimer-based nanoconstructions as delivery systems for short small interfering RNAs (siRNAs) and microRNAs at the cell and organism levels. It provides an overview of the structural features of dendrimers, indicating their advantages over other types of TNA transporters.


Identification of two novel mutations in the SLC45A2 gene in a Hungarian pedigree affected by unusual OCA type 4.
Oculocutaneous albinism (OCA) is a clinically and genetically heterogenic group of pigmentation abnormalities. OCA type IV (OCA4, OMIM 606574) develops due to homozygous or compound heterozygous mutations in the solute carrier family 45, member 2 (SLC45A2) gene. This gene encodes a membrane-associated transport protein, which regulates tyrosinase activity and, thus, melanin content by changing melanosomal pH and disrupting the incorporation of copper into tyrosinase. Here we report two Hungarian siblings affected by an unusual OCA4 phenotype. After genomic DNA was isolated from peripheral blood of the patients, the coding regions of the SLC45A2 gene were sequenced. In silico tools were applied to identify the functional impact of the newly detected mutations. Direct sequencing of the SLC45A2 gene revealed two novel, heterozygous mutations, one missense (c.1226G > A, p.Gly409Asp) and one nonsense (c.1459C > T, p.Gln437*), which were present in both patients, suggesting the mutations were compound heterozygous. In silico tools suggest that these variations are disease causing mutations. The newly identified mutations may affect the transmembrane domains of the protein, and could impair transport function, resulting in decreases in both melanosomal pH and tyrosinase activity. Our study provides expands on the mutation spectrum of the SLC45A2 gene and the genetic background of OCA4.


Antilymphocytic and immunosuppressive effects of Lantana camara leaves in rats.
Once daily administration of Lantana camara leaves juice at different dose levels (60, 300, 600 and 1500 mg/kg/day) for 14 days in rats resulted in alterations in various haemato- and biochemical parameters. Significant increase in blood urea nitrogen was observed with the doses of 600 and 1500 mg while significant increase in the relative weights of adrenals was observed at all the four dose levels. Total proteins, globulins, absolute lymphocyte count and per cent lymphocyte count were significantly decreased with 60, 600 and 1500 mg doses while a significant hypoglycemic effect was observed with 1500 mg only. Rats treated with 1500 mg dose did not exhibit any increase in alanine aminotransferase and aspartate aminotransferase activities or the alterations in relative kidney and liver weights. In another set of experiment, once daily oral administration of 1500 mg/kg/day for 14 days significantly inhibited the granulomatous tissue formation in rats and this effect was comparable to that of cyclophosphamide (10 mg/kg/day).


Factors Predicting the Final Diagnosis in Image-Guided Percutaneous Needle Biopsy for Suspected Spinal Tumors.
In cases of suspected spinal tumors on imaging studies, a biopsy is often necessary for establishing the diagnosis. Predictive factors for tumors or malignancies may help in scheduling biopsies or avoiding unnecessary ones. However, there have been few studies on determining these factors. We aimed to determine the factors associated with the final diagnosis in cases requiring spinal biopsy. This study included 117 patients who underwent image-guided (fluoroscopy- or computed tomography [CT]-guided) needle biopsy of the spine. Data on patient demographic, pathological diagnoses, and final diagnoses were retrospectively collected from the medical records. The imaging features and location of lesions were also evaluated on CT and magnetic resonance imaging. Furthermore, factors related to tumors or malignancies were analyzed. The diagnostic accuracy of biopsy was 94.0%, and there was no difference in the diagnostic accuracy between the fluoroscopic and CT-guided biopsies. Sixty-six and fifty-six patients were diagnosed with spinal tumors and malignant tumors, respectively. Multivariate analysis revealed that a history of malignant tumors and the presence of pedicle lesions and/or extravertebral lesions were related factors for both tumors or malignancy in the final diagnosis. These findings can help determine the necessity for or timing of biopsy in patients with suspected spinal tumors.


Complications Leading to Keratoplasty among Contact Lens Users and LASIK Patients: A 10-Year Cross-Sectional Analysis.
To determine the incidence and outcomes in patients who underwent penetrating keratoplasty (PK) resulting from complications related to contact lens (CL) use and laser in situ keratomileusis (LASIK) in a metropolitan area of the United States. Population data was obtained from the United States Census Bureau and the Centers for Disease Control. A retrospective, cross-sectional chart review was performed on all patients who underwent keratoplasty in a specific metropolitan geographic area over a ten-year period. The main outcome was best-corrected visual acuity (BCVA) at 2 years in patients who underwent PK secondary to complications related to CL use and LASIK. The secondary outcome was the relative risk of undergoing PK secondary to a complication related to CL use versus LASIK. The study's geographic area had 46,545 CL users in one or both eyes during any given year and 10,285 patients who underwent LASIK in one or both eyes during the study interval. There were 24 CL users (0.52 per 1,000) and 3 post-LASIK patients (0.29 per 1,000) who underwent PK secondary to complications during the study interval (OR 1.77 [0.53-5.87, 95% CI]; p=0.35). BCVA at 2 years was 1.45 [1.0-1.90] logMAR (20/564 Snellen) in the CL using cohort and 0.07 [-1.19-1.33] logMAR (20/23 Snellen) in post-LASIK cohort following PK (p=0.04). Patients who underwent PK secondary to complications related to CL use had worse visual outcomes at 2 years compared to those related to LASIK. Complications leading to PK were rare in both cohorts, but the incidence of undergoing PK secondary to CL use trended higher than LASIK.


Depressive symptoms are more strongly related to executive functioning and episodic memory among African American compared with non-Hispanic White older adults.
We examined whether the reserve capacity model can be extended to cognitive outcomes among older African Americans. Two hundred and ninety-two non-Hispanic Whites and 37 African Americans over age 54 participated in the normative study for the NIH Toolbox for the Assessment of Neurological and Behavioral Function. Multiple-group path analysis showed that associations between depressive symptoms and cognition differed by race, independent of age, education, reading level, income, health, and recruitment site. Depressive symptoms were associated with slowed processing speed among Whites and worse task-switching, inhibition, and episodic memory among African Americans. African Americans may be more vulnerable to negative effects of depression on cognition than non-Hispanic Whites. Further research is needed to explicate the psychological and neurobiological underpinnings of this greater vulnerability.


Immunohistochemical analysis of nuclear survivin expression in esophageal squamous cell carcinoma.
Despite advances in the treatment of esophageal carcinoma, the prognosis for this disease remains poor. Therefore, it is important to obtain a better understanding of the molecular basis of esophageal carcinogenesis. The purpose of this study was to clarify the roles of survivin in esophageal squamous cell carcinoma (ESCC). One hundred 22 ESCC surgical specimens resected from 1989 to 1999 were examined. Survivin expression was assessed by immunohistochemistry. Tumor cells were considered survivin-positive if the immunoreactivity was confined to the nucleus, and a scoring method was applied. Survivin-positive immunostaining was detected in 68 patients (56%). There was a significant association between survivin expression and pN (P = 0.0472). Moreover, the overall survival rate was worse in patients with survivin-positive tumors than in patients with survivin-negative tumors (P = 0.0189). The overexpression of survivin was associated with the overall survival rate and poor prognosis in patients with ESCC. Survivin may be targeted during cancer therapy because of its selective expression in malignant tissue.


Pharmacokinetic differences of mifepristone between sexes in animals.
Mifepristone (RU486) is developed originally as a contraceptive used by hundreds of millions of women world-wide, and also reported as a safe and long-term psychotic depressant, or as a cancer chemotherapeutic agent used by both sexes. In our preliminary study aimed at developing mifepristone as a cancer metastatic chemopreventive, we coincidentally observed that blood mifepristone concentrations in female rats seem to be higher than those in male ones post administration. To substantiate if the pharmacokinetic differences between sexes exist, we established a fast UPLC-MS/MS method to determine mifepristone concentrations in plasma, and analyzed blood concentrations of mifepristone over time in rats and dogs of both sexes. Mifepristone in plasma or incubation liquid was recovered by liquid-liquid extraction using 1 mL of ethyl acetate. Chromatographic separation was performed on a C18 column at 35 °C, with a gradient elution consisting of methanol and water containing 0.1% (v/v) formic acid at a flow rate of 0.3 mL/min. And pharmacokinetic parameters such as elimination half-life, and mean residence time were calculated by using the non-compartmental pharmacokinetics data analysis software. In this work, administrations of mifepristone to rats and beagle dogs revealed that the plasma concentrations of mifepristone (AUC, Cmax) were significantly higher (P < 0.05) in females than that in males. In vitro liver microsomal incubation experiments showed that the metabolic rate of mifepristone in males was higher than that in females, which was consistent with the results of in vivo experiments. In general, we first found the sex-related differences about pharmacokinetic properties of mifepristone and revealed the metabolism difference of hepatic microsomal enzyme is the main reason.


Membrane solubility parameter and in situ release of theophylline.
The solubility of theophylline in polyethylene glycol 400-water binary mixtures was analyzed in terms of solute-solvent interaction using the solubility parameter principle of Hildebrand. Preliminary in situ (rat gut permeation) studies with the solvent mixtures having varying polarity as expressed by the solubility parameter showed that (a) the more alike the solubility parameter of theophylline (delta 2 = 14, as obtained from solubility studies) and the solubility parameter of the solvent mixture, the greater was the attraction of solvent for the drug (b) sufficient similarity must exist between the solubility parameter of theophylline and that of the intestinal mucosa to promote bioabsorption, and (c) from the above, it follows that there exists a competition between solvent and rat gut membrane for the drug. The solubility parameter of the solvent vehicle must be such that it does not impede absorption of the molecules into the intestinal mucosa.


The many faces of surgical margins.
Most variables associated with survival after cancer surgery are fixed when we see the patient. One variable over which we have control is the surgical margin. We begin by reviewing the definitions of a positive surgical margin, and then explore potential inaccuracies in obtaining a biopsy and reasons for recurrence after obtaining a free surgical margin. Research in improving the diagnostic accuracy of surgical margins is discussed. Finally, the prognostic significance of surgical margins is reviewed.


Human immunodeficiency virus and leprosy.
In summary, clinical leprosy does not appear to be more frequent in HIV-positive than in HIV-negative people in areas where both infections are endemic. There is no evidence that the paucibacillary to multibacillary distribution of patients is altered by HIV infection. There are reports that neuritis is more severe in co-infected people, and that reversal reactions (or, at least, new lesions) may be more frequent after therapy; but these reports are either poorly documented or poorly controlled. In several ways, this lack of expected leprosy is similar to the patterns of other low-virulent infections in HIV-positive patients in the tropics, such as MAC infection, Pneumocystis carinii and cytomegalovirus infection. Despite the presence of the agents in the environment and/or in the human host, they are infrequently encountered clinicopathologically. This is in marked contrast to their importance in industrialized countries. One explanation, as yet unproven, is that HIV-positive patients in the tropics do not live long enough in states of severe immunosuppression to develop these infections. Perhaps the same applies to M. leprae infection, whose incubation period can be measured in decades and whose clinical course may evolve over years, in contrast to the common reactivation of latent, virulent M. tuberculosis infection with its high morbidity. There may also be an analogy between leprosy and infection with Plasmodium falciparum, Strongyloides stercoralis and Entamoeba histolytica: these infections are controlled (or at least influenced) by cell-mediated immunity and, theoretically, should be more frequent in HIV-positive people than HIV-negatives. In fact they are not, and leprosy may be regarded as another 'missing infection in AIDS'.(ABSTRACT TRUNCATED AT 250 WORDS)


The axonal transport motor kinesin-2 navigates microtubule obstacles via protofilament switching.
Axonal transport involves kinesin motors trafficking cargo along microtubules that are rich in microtubule-associated proteins (MAPs). Much attention has focused on the behavior of kinesin-1 in the presence of MAPs, which has overshadowed understanding the contribution of other kinesins such as kinesin-2 in axonal transport. We have previously shown that, unlike kinesin-1, kinesin-2 in vitro motility is insensitive to the neuronal MAP Tau. However, the mechanism by which kinesin-2 efficiently navigates Tau on the microtubule surface is unknown. We hypothesized that mammalian kinesin-2 side-steps to adjacent protofilaments to maneuver around MAPs. To test this, we used single-molecule imaging to track the characteristic run length and protofilament switching behavior of kinesin-1 and kinesin-2 motors in the absence and presence of 2 different microtubule obstacles. Under all conditions tested, kinesin-2 switched protofilaments more frequently than kinesin-1. Using computational modeling that recapitulates run length and switching frequencies in the presence of varying roadblock densities, we conclude that kinesin-2 switches protofilaments to navigate around microtubule obstacles. Elucidating the kinesin-2 mechanism of navigation on the crowded microtubule surface provides a refined view of its contribution in facilitating axonal transport.


Quantitative phase imaging for characterization of single cell growth dynamics.
Quantitative phase imaging (QPI) has emerged as an indispensable tool in the field of biomedicine, offering the ability to obtain quantitative maps of phase changes due to optical path length delays without the need for contrast agents. These maps provide valuable information about cellular morphology and dynamics, unperturbed by the introduction of exogenous substances. In this review, a summary of recent studies that have focused on elucidating the growth dynamics of individual cells using QPI is presented. Specifically, investigations into cellular changes occurring during mitosis, the differentiation of cellular organelles, the assessment of distinct cell death processes (i.e., apoptosis, necrosis, and oncosis) and the precise measurement of live cell temperature are explored. Furthermore, the captivating applications of QPI in theragnostics, where its potential for transformative impact is prominently showcased, are highlighted. Finally, the challenges that need to be overcome for its wider adoption and successful integration into biomedical research are outlined.


Translocation of Phthalates From Food Packaging Materials Into Minced Beef.
There has been increased concern regarding the potential human health risks associated with exposure to phthalates. Research indicates that food intake is the most critical exposure pathway for phthalates. This study aimed to investigate packaged beef samples for the presence of dimethyl terephthalate (DMTP), di-n-butyl phthalate (DnBP), and diisooctyl phthalate (DiOP) and to assess their translocation from the common form of food packaging procured from various Saint-Petersburg and Leningrad region shops. The packaging samples include paper and different types of plastic. Phthalates were extracted by dichloromethane and analyzed by gas chromatography coupled with mass spectrometry (GC-MS). While DnBP had the highest mean values in beef from 34.5 to 378.5 μg·kg-1, DiOP displayed the lowest mean values from LOD to 37 μg·kg-1. The larger contact area and the presence of distributed fat on the surface of the minced meat resulted in significantly higher phthalate translocation than beef slices. Further, DMTP was not detected in any samples. However, the examined food packages do not meet the requirements of Russian, EU and USA legislation, as DnBP migrates to meat. Calculated maximum DnBP daily intake of 0.167 μg·kg-1·day-1 for chilled minced beef in vacuum packaging did not exceed tolerable daily intake (TDI) level. The most alarming results are concerning the phthalates presence in beef farmed in the Leningrad region and not subjected to any plastic packaging. A full-scale study is warranted to determine the pathways and sources of phthalates migration in the food chain.


[An update on parathyroid surgery].
The operative management of patients with hyperparathyroidism is controversial. High rates of persistent hypercalcemia and postoperative hypoparathyroidism are seen in multiple hyperplasia and bilateral neck exploration. Patients undergoing unilateral neck exploration with removal of a single parathyroid adenoma have a rapid clearance of PTH which declines within hours after surgery. There is a sensitive immunometric assay (IRMA) for the intact molecule which demonstrates a decline in 15 minutes during surgery. Intraoperative measurement of PTH may be complementary to surgical skill and histopathologic information and may modulate extension of neck exploration.


Case report: An unusual presentation of acute promyelocytic leukemia in a middle aged female mimicking dengue infection.
Acute promyelocytic leukemia (APL) is an uncommon subtype of acute myeloid leukemia (AML). M3v phenotype is a less common presentation of APL and these patients usually present with leukocytosis and abnormal promyelocytes that are characterized by sparse granulation and are less likely to have faggot cells with multiple Auer rods. Distinguishing M3v phenotype from acute febrile illness can be challenging as the diagnosis relies on examination of peripheral smear. Fifty-seven-year-old female who presented after recent trip to Dominican Republic for high grade fever and gum bleeding. She was exposed to patients with Dengue fever during her stay. At presentation, patient had leukocytosis, thrombocytopenia, and urinalysis showing bacteria and white cell. She was started on treatment for urinary tract infection. Patient remained febrile and thrombocytopenia worsened. On day 2, flow cytometry of the peripheral smear showed 43% medium sized blasts. Fluorescence in situ hybridization was positive for promyelocytic leukemia/retinoic acid receptor alpha. The patient was diagnosed with APL. Patient was started on treatment with all-trans retinoic acid and arsenic trioxide along with supportive care OUTCOMES:: Patient had a favorable clinical response and her symptoms subsided. Flow cytometry of the peripheral smear is key to diagnosis of suspected APL. One must maintain high suspicion for this life-threatening condition as early diagnosis saves lives.


Acetylcholine-induced coronary spasm in patients with unobstructed coronary arteries is associated with elevated concentrations of soluble CD40 ligand and high-sensitivity C-reactive protein.
To assess whether epicardial and microvascular coronary artery spasm in response to acetylcholine (ACH) is associated with markers of inflammation, platelet stimulation, and endothelial activation in patients with angina and unobstructed coronary arteries. Patients with angina pectoris despite angiographically normal coronary arteries represent a diagnostic and therapeutic challenge. Both impaired coronary microvascular dilatory responses as well as diffuse distal epicardial and microvascular coronary artery spasm have been described as possible pathogenic mechanisms. Although inflammation has been proposed to play a pathogenic role in angina, an association between ACH-induced coronary vasospasm and inflammation in Caucasians has not been reported previously in this context. We assessed 62 consecutive patients (26 men, age 60±10 years) with chest pain despite angiographically unobstructed coronary arteries (<50% stenosis) who underwent intracoronary ACH testing for the diagnosis of coronary artery spasm. High-sensitivity C-reactive protein (hs-CRP), e-selectin, neopterin, and sCD40L concentrations were measured in all patients before ACH testing. The ACH test was considered to be 'positive' in the presence of (a) angina and at least 75% coronary diameter reduction (epicardial coronary artery spasm) or (b) ischemic ST-shifts and angina in the absence of epicardial spasm (microvascular spasm). Eight patients without angina pectoris served as a control group. The ACH test was positive in 48 patients (77%). Twenty-seven patients had epicardial spasm (56%) and 21 patients had microvascular spasm (44%). Epicardial spasm was diffuse in 26 patients (96%) and focal in one patient (4%). Elevated hs-CRP, e-selectin, and sCD40 ligand concentrations were significantly (P≤0.05) associated with a positive ACH-test response. Hs-CRP (odds ratio 1.54, confidence interval 1.02-2.33, P=0.04) and sCD40 ligand (odds ratio 1.001, confidence interval 1.00-1.001, P=0.003) were predictors for a positive ACH test on multivariate analysis. None of the patients in the control group developed epicardial or microvascular spasm during ACH testing. Epicardial and microvascular coronary spasm in response to ACH correlate significantly with hs-CRP and sCD40 ligand concentrations in patients with angina pectoris and angiographically unobstructed coronary arteries. These results suggest that an association exists between inflammation and coronary artery spasm in patients with angina pectoris despite unobstructed coronary arteries and studies are needed to explore the mechanisms underlying this association.


Detection and quantification of noroviruses in shellfish.
Noroviruses (NoVs) are the most common viral agents of acute gastroenteritis in humans, and high concentrations of NoVs are discharged into the environment. As these viruses are very resistant to inactivation, the sanitary consequences are contamination of food, including molluscan shellfish. There are four major problems with NoV detection in shellfish samples: low levels of virus contamination, the difficulty of efficient virus extraction, the presence of interfering substances that inhibit molecular detection, and NoV genetic variability. The aims of this study were to adapt a kit for use with a method previously shown to be efficient for detection of NoV in shellfish and to use a one step real-time reverse transcription-PCR method with addition of an external viral control. Comparisons of the two methods using bioaccumulated oysters showed that the methods reproducibly detected similar levels of virus in oyster samples. Validation studies using naturally contaminated samples also showed that there was a good correlation between the results of the two methods, and the variability was more attributable to the level of sample contamination. Magnetic silica very efficiently eliminated inhibitors, and use of extraction and amplification controls increased quality assurance. These controls increased the confidence in estimates of NoV concentrations in shellfish samples and strongly supported the conclusion that the results of the method described here reflected the levels of virus contamination in oysters. This approach is important for food safety and is under evaluationfor European regulation.


De novo methylation of MMLV provirus in embryonic stem cells: CpG versus non-CpG methylation.
DNA methyltransferases, Dnmt3a and Dnmt3b, are required for de novo methylation in embryonic stem (ES) cells and postimplantation embryos. However, the mechanism of de novo methylation is largely unknown. In this study, we have analyzed the sequence specificity of Dnmt3a and Dnmt3b during de novo methylation of murine Moloney leukemia virus provirus DNA in virus-infected ES cells. Provirus DNA from infected wild-type (J1), Dnmt1-/- (c/c), and Dnmt3a3b-/- (3a3b-/-) ES cells were analyzed using the bisulfite sequencing method. We demonstrate that Dnmt3 enzymes methylate predominantly CpG sites in vivo and confirm that Dnmt3 enzymes, but not Dnmt1, are responsible for de novo methylation. However, the sequence context and CpG density do not appear to influence de novo methylation, though strand bias is detectable. Interestingly, non-CpG methylation is detected as a component of de novo methylation. CpA methylation was detected at approximately 1.4% of all sites in J1 and approximately 1.0% in c/c, but only approximately 0.2% in 3a3b-/-. Few methylated CpT or CpC sites were detected. Similar results from nearest neighbor analysis of global endogenous methylation levels indicated a correlation between Dnmt3a and Dnmt3b presence and CpA methylation. These results demonstrate that the Dnmt3 enzymes methylate predominantly CpG sites and at a low frequency CpA sites with no apparent sequence preferences.


DNA release as a direct measure of microbial killing. I. Serum bactericidal activity.
A new method for quantification of microbial killing is presented, based upon the assumption that release of DNA from an organism can be taken as direct evidence of cell death. The assay was applied to measurement of serum bactericidal activity. A "serum-sensitive" Escherichia coli, whose DNA was pre-labeled with [14C] thymidine, released 95 to 100% of its radioactivity upon exposure to human serum for 120 min. This was accompanied by a fall in viability with less than 0.1% of the bacteria surviving. Heating serum to 56 degrees C for 30 min completely abolished both DNA release and killing. Normal serum did not release DNA from a "serum-resistant". Salmonella typhimurium whereas exposure to ampicillin caused both significant killing and DNA release. This assay is highly specific, sensitive, and rapid.


Medical and physical therapy of temporomandibular joint disk displacement without reduction.
The objective of this study was to determine the influence of medical and physical therapy on long-term treatment outcome in 72 patients with anterior disk displacement without reduction. Patients were treated solely with occlusal splints (group I), with splints and supplementary medical therapy (group II), with splints and physical therapy (group III) or with splints, medical, and physical therapy (group IV). After therapy, the maintenance of improvement was objectively and subjectively assessed with an extensive clinical examination and a postal questionnaire. The percentage of pain free patients after therapy was 76% in group I, 88% in group II, 43% in group III, and 65% in group IV. There was a statistically significant higher increase of maximum jaw opening after therapy in group II than in the control groups (p<0.05). The improvement in mouth opening came to 9.7 mm in group I, 14.5 mm in group II, 7.3 mm in group III, and 11.2 mm in group IV. Medical therapy seems to have a positive influence on the treatment outcome of patients with anterior disk displacement without reduction.


Configurational factors in the perception of unfamiliar faces.
Young et al (1987) have demonstrated that the juxtaposition of top and bottom halves of different faces produces a powerful impression of a novel face. It is difficult to isolate perceptually either half of the 'new' face. Inversion of the stimulus, however, makes this task easier. Upright chimeric faces appear to evoke strong and automatic configurational processing mechanisms which interfere with selective piecemeal processing. In this paper three experiments are described in which a matching paradigm was used to show that Young et al's findings apply to unfamiliar as well as to familiar faces. The results highlight the way in which minor procedural differences may alter the way in which subjects perform face-recognition tasks.


Ex vivo genome-wide RNAi screening of the Drosophila Toll signaling pathway elicited by a larva-derived tissue extract.
Damage-associated molecular patterns (DAMPs), so-called "danger signals," play important roles in host defense and pathophysiology in mammals and insects. In Drosophila, the Toll pathway confers damage responses during bacterial infection and improper cell-fate control. However, the intrinsic ligands and signaling mechanisms that potentiate innate immune responses remain unknown. Here, we demonstrate that a Drosophila larva-derived tissue extract strongly elicits Toll pathway activation via the Toll receptor. Using this extract, we performed ex vivo genome-wide RNAi screening in Drosophila cultured cells, and identified several signaling factors that are required for host defense and antimicrobial-peptide expression in Drosophila adults. These results suggest that our larva-derived tissue extract contains active ingredients that mediate Toll pathway activation, and the screening data will shed light on the mechanisms of damage-related Toll pathway signaling in Drosophila.


Atomic Bose and Anderson glasses in optical lattices.
An ultracold atomic Bose gas in an optical lattice is shown to provide an ideal system for the controlled analysis of disordered Bose lattice gases. This goal may be easily achieved under the current experimental conditions by introducing a pseudorandom potential created by a second additional lattice or, alternatively, by placing a speckle pattern on the main lattice. We show that, for a noncommensurable filling factor, in the strong-interaction limit, a controlled growing of the disorder drives a dynamical transition from superfluid to Bose-glass phase. Similarly, in the weak interaction limit, a dynamical transition from superfluid to Anderson-glass phase may be observed. In both regimes, we show that even very low-intensity disorder-inducing lasers cause large modifications of the superfluid fraction of the system.


Ether for Wound Debridement of Wild Maggots.
Myiasis is defined as an infestation of the organs or tissues of live vertebrates (humans or animals) by dipterous larvae, commonly referred to as maggots. Wound myiasis is a common presentation in the emergency department. The presence of wild maggots and larvae in wounds is concerning for subsequent myiasis-induced infections and complications. Proper extraction of wild maggots and larvae is required to avoid complications that may occur with wound myiasis. Surgical debridement is often utilized to extract wild maggots and larvae, but can be painful and time-consuming. There is limited literature available on alternative approaches for removal of wild maggots and larvae. We present a case of lower extremity wound myiasis that had wild maggots and larvae successfully removed with no complications using ether. This procedure is a novel, noninvasive, and pain-free way to extract wild maggots and larvae from wound myiasis.


The catalytic effect of water, water dimers and water trimers on H2S + (3)O2 formation by the HO2 + HS reaction under tropospheric conditions.
In this article, the reaction mechanisms of H2S + (3)O2 formation by the HO2 + HS reaction without and with catalyst X (X = H2O, (H2O)2 and (H2O)3) have been investigated theoretically at the CCSD(T)/6-311++G(3df,2pd)//B3LYP/6-311+G(2df,2p) level of theory, coupled with rate constant calculations by using conventional transition state theory. Our results show that in the presence of catalyst X (X = H2O, (H2O)2 and (H2O)3) into the channel of H2S + (3)O2 formation, the reactions between the SH radical and HO2(H2O)n (n = 1-3) complexes are more favorable than the corresponding reactions of the HO2 radical with HS(H2O)n (n = 1-3) complexes due to the lower barrier of the former reactions and the higher concentrations of HO2(H2O)n (n = 1-3) complexes. Meanwhile, the catalytic effect of water, water dimers and water trimers is mainly taken from the contribution of a single water vapor molecule, since the total effective rate constant of HO2H2O + HS and H2OHO2 + HS reactions was, respectively, larger by 7-9 and 9-12 orders of magnitude than that of SH + HO2(H2O)2 and SH + HO2(H2O)3 reactions. Besides, the enhancement factor of water vapor is only 0.37% at 240 K, while at high temperatures, such as 425 K, the positive water vapor effect is enhanced up to 38.00%, indicating that at high temperatures the positive water effect is obvious under atmospheric conditions. Overall, these results show how water and water clusters catalyze the gas phase reactions under atmospheric conditions.


[The promoting research of phospholipase C epsilon-1 on nasal Th2 cell polarization].
Phospholipase C epsilon-1 (PLCE1) is a phospholipase C isoenzyme encoded by PLCE1 gene, and has more complicated molecular structure and function than other subtypes. Phospholipase C epsilon-1 is accepted the dual regulation by the upstream G proteins and GTP enzymes of Ras family. The downstream signal of PLCE1 is not only cause the Ca2+ flow and protein kinase C(PKC) activation, but also can be used as the GTP enzyme guanylic acid conversion factor of Ras superfamily, so as to regulate the expression of certain genes, adjusting cell growth and differentiation processes. PLCE1 plays a very important role in the signal transduction in the regulation of cell growth, differentiation, proliferation and apoptosis. Previous studies showed that phospholipase C epsilon-1 played an important role in the development of malignant tumors (especially the digestive tumors), heart disease, nephrotic syndrome and other diseases, but there are some questions about the mechanisms of PLCE1 involved in allergic rhinitis, this article will make an overview about PLCE1 promotes allergic rhinitis CD4+ T cells differentiate to Th2 cells by PKC-NF-κB pathway and Ras-MAPK pathway.


TRIENNIAL GROWTH AND DEVELOPMENT SYMPOSIUM: Dedifferentiated fat cells: Potential and perspectives for their use in clinical and animal science purpose.
An increasing body of evidences has demonstrated the ability of the mature adipocyte to dedifferentiate into a population of proliferative-competent cells known as dedifferentiated fat (DFAT) cells. As early as the 1970s, in vitro studies showed that DFAT cells may be obtained by ceiling culture, which takes advantage of the buoyancy property of lipid-filled cells. It was documented that DFAT cells may acquire a phenotype similar to mesenchymal stem cells and yet may differentiate into multiple cell lineages, such as skeletal and smooth muscle cells, cardiomyocytes, osteoblasts, and adipocytes. Additionally, recent studies showed the ability of isolated mature adipocytes to dedifferentiate in vivo and the capacity of the progeny cells to redifferentiate into mature adipocytes, contributing to the increase of body fatness. These findings shed light on the potential for use of DFAT cells, not only for clinical purposes but also within the animal science field, because increasing intramuscular fat without excessive increase in other fat depots is a challenge in livestock production. Knowledge of the mechanisms underlying the dedifferentiation and redifferentiation of DFAT cells will allow the development of strategies for their use for clinical and animal science purposes. In this review, we highlight several aspects of DFAT cells, their potential for clinical purposes, and their contribution to adipose tissue mass in livestock.


Structural relaxation in charged metal surfaces and cluster ions.
Space-charge layers can noticeably affect the properties of metal electrode surfaces, for instance by modifying the surface dielectric response or indirectly via the induced atomic relaxations. While there are efforts to exploit this concept for designing novel functional nanomaterials, the underlying microscopic processes are poorly understood. Here, we report on a density functional theory (DFT) study of atomic relaxation in Au cluster ions comprising up to 309 atoms. Suitable averages over atomic displacements respond to charging consistent with experimental observation on macroscopic Au single-crystal surfaces. Moreover, the overall DFT response is also consistent with predictions of a simple phenomenological model. Motivated by our observations, we propose a scenario in which the surface relaxation ("stretch") results from out-of-plane Hellman-Feynman forces exerted on the surface atoms by the excess charge, and where the in-plane surface stress represents essentially an elastic transverse contraction tendency of the surface layer in response to stretch.


Meta-analytic evidence for downregulation of the amygdala during working memory maintenance.
The amygdala is a region critically implicated in affective processes. Downregulation of the amygdala is one of the hallmarks of successful emotion regulation. Top-down inhibition of the amygdala is thought to involve activation of the executive control network. This reciprocal relationship, however, is not exclusive to explicit emotion regulation. It has been noted that any cognitively demanding task that activates executive control network may downregulate the amygdala, including a standard working memory task. Such downregulation is likely established in a load-dependent fashion with more cognitive demand leading to stronger deactivation. Using a coordinate-based meta-analysis, we examined whether a standard working memory task downregulates the amygdala similarly to cognitive reappraisal. We found that a standard 2-back working memory task indeed systematically downregulates the amygdala and that deactivated clusters strongly overlap with those observed during a cognitive reappraisal task. This finding may have consequences for the interpretation of the underlying mechanism of cognitive reappraisal: amygdala downregulation may be related to the cognitively demanding nature of reappraisal and not per se by the act of the reappraisal itself. Moreover, it raises the possibility of applying working memory tasks in clinical settings as an alternative emotion regulation strategy.


Successful treatment of a rectal anastomotic stenosis by transanal endoscopic microsurgery (TEM) using the contact Nd:YAG laser.
We report the advantage of employing transanal endoscopic microsurgery (TEM) using the contact Nd:YAG laser for the treatment of a rectal anastomotic stenosis. A 72-year-old woman was admitted to our hospital with a postoperative rectal anastomotic stenosis. Twenty months prior to admission, the patient underwent a low anterior resection for the treatment of the rectal cancer using an EEA stapling device. A barium enema and colonoscopy revealed a rectal stenosis, 0.8-cm diameter. This stenosis was at the anastomotic site, approximately 4.0 from the dental line. An endoscopic treatment was performed transanally using the contact Nd:YAG laser. The stenotic rectal wall was fulgurated or vaporized completely. There were no intraoperative or postoperative complications. We concluded that TEM appears to be a safe and minimally invasive procedure. Furthermore, the contact Nd:YAG laser is very effective in treating the gastrointestinal stenotic area. To our knowledge, this is the first successful report of this novel procedure.


Two sites in the MAPT region confer genetic risk for Guam ALS/PDC and dementia.
Unusual forms of amyotrophic lateral sclerosis (ALS-G), Parkinsonism dementia complex (PDC-G) and Guam dementia (GD) are found in Chamorros, the indigenous people of Guam. Neurofibrillary tangles composed of hyperphosphorylated tau are a neuropathologic feature of these closely related disorders. To determine if variation in the gene that encodes microtubule-associated protein tau gene (MAPT) contributes to risk for these disorders, we genotyped nine single nucleotide polymorphism (SNP) sites and one insertion/deletion in the 5' end of MAPT in 54 ALS-G, 135 PDC-G, 153 GD and 258 control subjects, all of whom are Chamorros. Variation at three SNPs (sites 2, 6 and 9) influenced risk for ALS-G, PDC-G and GD. SNP2 acts through a dominant mechanism and is independent of the risk conferred by SNPs 6 and 9, the latter two acting by a recessive mechanism. Persons with the high-risk SNP6 and SNP9 AC/AC diplotype had an increased risk of 3-fold [95% confidence interval (CI)=1.10-8.25] for GD, 4-fold (95% CI=1.40-11.64) for PDC-G and 6-fold (95% CI=1.44-32.14) for ALS-G, compared to persons with other diplotypes after adjusting for SNP2. Carriers of the SNP2 G allele had an increased risk of 1.6-fold (95% CI=1.00-2.62) for GD, 2-fold (95% CI=1.28-3.66) for PDC-G, and 1.5-fold (95% CI=0.74-3.00) for ALS-G, compared to non-carriers after adjusting for SNPs 6 and 9. Others have shown that SNP6 is also associated with risk for progressive supranuclear palsy. These two independent cis-acting sites presumably influence risk for Guam neuro-degenerative disorders by regulating MAPT expression.


Oral administration of NGF synthesis stimulators recovers reduced brain NGF content in aged rats and cognitive dysfunction in basal-forebrain-lesioned rats.
Nerve growth factor (NGF) plays an important role in the survival and maintenance of cholinergic neurons in the central neuronal system. Since this factor does not cross the blood-brain barrier and is easily metabolized by peptidases when administered peripherally, it can be used for medical treatment only when directly injected into the brain. We report here that repeated oral administration of the stimulators for the NGF synthesis, idebenone and propentofylline, produced a significant recovery of the reduced NGF content in the frontal and parietal cortices of aged rats. These compounds also improved deficits of performance in water maze, passive avoidance and habituation tasks in basal-forebrain-lesioned rats. These results suggest that the use of the stimulators for the NGF synthesis may provide a therapeutic approach to cholinergic dysfunction.


International role of the perioperative nurse.
Perioperative nursing extends outside of our operating room environments, not only within our hospitals, but also internationally, across borders. Those perioperative nurses who are interested in international care can easily become involved with a project within their own city and state. The challenge is the commitment and the effort.


Multiplexed single-molecule measurements with magnetic tweezers.
We present a method for performing multiple single-molecule manipulation experiments in parallel with magnetic tweezers. We use a microscope with a low magnification, and thus a wide field of view, to visualize multiple DNA-tethered paramagnetic beads and apply an optimized image analysis routine to track the three-dimensional position of each bead simultaneously in real time. Force is applied to each bead using an externally applied magnetic field. Since variations in the field parameters are negligible across the field of view, nearly identical manipulation of all visible beads is possible. However, we find that the error in the position measurement is inversely proportional to the microscope's magnification. To mitigate the increased error caused by demagnification, we have developed a strategy based on tracking multiple fixed beads. Our system is capable of simultaneously manipulating and tracking up to 34 DNA-tethered beads at 60 Hz with approximately 1.5 nm resolution and with approximately 10% variation in applied force.


Local arterial wall drug delivery using balloon catheter system.
Balloon-based drug delivery systems allow localized application of drugs to a vascular segment to reduce neointimal hyperplasia and restenosis. Drugs are coated onto balloons using excipients as drug carriers to facilitate adherence and release of drug during balloon inflation. Drug-coated balloon delivery system is characterized by a rapid drug transfer that achieves high drug concentration along the vessel wall surface, intended to correspond to the balloon dilation-induced vascular injury and healing processes. The balloon catheter system allows homogenous drug delivery to the vessel wall, such that the drug release per unit surface area is kept constant along balloons of different lengths. Optimization of the balloon coating matrix is essential for efficient drug transfer and tissue retention until the artery remodels to a normal set point. Challenges in the development of balloon-based drug delivery to the arterial wall include finding suitable excipients for drug formulation to enable drug release to a targeted lesion site effectively, maintain coating integrity during transit, prolong tissue retention and reduce particulate generation. This review highlights various factors involved in the successful design of balloon-based delivery systems, including drug release kinetics, matrix coating transfer, transmural drug partitioning, dissolution rate and release of unbound active drug.


Effect of Longitudinal Variation of Vocal Fold Inner Layer Thickness on Fluid-Structure Interaction During Voice Production.
A three-dimensional fluid-structure interaction computational model was used to investigate the effect of the longitudinal variation of vocal fold inner layer thickness on voice production. The computational model coupled a finite element method based continuum vocal fold model and a Navier-Stokes equation based incompressible flow model. Four vocal fold models, one with constant layer thickness and the others with different degrees of layer thickness variation in the longitudinal direction, were studied. It was found that the varied thickness resulted in up to 24% stiffness reduction at the middle and up to 47% stiffness increase near the anterior and posterior ends of the vocal fold; however, the average stiffness was not affected. The fluid-structure interaction simulations on the four models showed that the thickness variation did not affect vibration amplitude, glottal flow rate, and the waveform related parameters. However, it increased glottal angles at the middle of the vocal fold, suggesting that vocal fold vibration amplitude was determined by the average stiffness of the vocal fold, while the glottal angle was determined by the local stiffness. The models with longitudinal variation of layer thickness consumed less energy during the vibrations compared with the constant layer thickness one.


Effect of acute 60Co-gamma-irradiation on the in vivo lipid peroxidation in experimental animals.
The effect of sublethal (6.0 Gy) and LD50/30 (9.0 Gy) doses of 60Co-gamma-irradiation on malondialdehyde (MDA) level was studied in rats. The findings suggest that in the organs investigated (brain, liver, spleen, kidney, testicle, stomach, small intestines) acute 60Co-gamma-irradiation increased the formation of MDA, the main product of lipid peroxidation, in a time-related manner to an extent characteristic of the organ investigated. Differences in the degree and temporal development of the changes allow some conclusions as to the radiosensitivity of individual organs. On this basis it can be assumed that the increase in MDA level caused by irradiation considerably contributes to the development of certain symptoms of radiation sickness.


Beyond purely ethical understanding of responsibility: a phenomenological approach.
The assumption that moral responsibility is a constitutive feature of the human person could almost be considered as the raison d'être of ethics and consequently of bioethics. In fact, the concept of"common morality", which is becoming ever more popular within the mainstream of bioethics, assumes that certain, even if elementary, experience of moral responsibility (and understanding of it) is shared by all the people of every cultural (moral) tradition. Although the phenomenon of (moral) responsibility is seen as a moral question, its ultimate explanation needs reflection which goes beyond purely ethical considerations. The phenomenological ontology as elaborated by R. Ingarden is accepted as the theoretical background of deliberations in this study. Methods of reasoning appropriate to this approach to phenomenology are used. The phenomenon of human responsibility is analyzed--according to Ingarden's suggestion--from the point of view of the four "situations" in which responsibility appears: (I) to be responsible for something, (II) to accept responsibility for something, (III) to be called to responsibility for something, and (IV) to be acting responsibly. The ontological foundation of responsibility in each of these situations is then inquired. Ingarden's philosophical reflection leads to discovery of the ultimate ontological foundation of responsibility in: (I) objective axiology, (II) structure of an acting subject, and (III) a causality structure of reality. We discuss whether Ingarden's analyses are coherent with the concept of "common morality". Although R. Ingarden is best known, especially to English-speaking readers, for his work in aesthetics, it should be emphasized that his phenomenological ontology can provide interesting insights into the understanding of the concept of responsibility which is fundamental to ethics/bioethics.


Influence of Clostridium botulinum C2 toxin on Fc epsilonRI-mediated secretion and tyrosine phosphorylation in RBL cells.
We studied the effects of the binary Clostridium botulinum C2 toxin on stimulated [3H]serotonin release and protein tyrosine phosphorylation in RBL 2H3 hm1 cells. Actin was specifically ADP-ribosylated by C2 toxin in intact cells resulting in a 2-3 fold increase in antigen- or calcium ionophore (A23187)-induced degranulation. The effects of C2 toxin were time- and concentration-dependent. Toxin treatment, which dramatically changes the morphology of RBL cells, was not sufficient to induce mediator release in the absence of activators of secretion. Antigen- and A23187-stimulated tyrosine phosphorylation of 60-80 kDa and 110-120 kDa proteins was reduced or blocked after C2 toxin incubation. Treatment of RBL cells with the tyrosine phosphatase inhibitor pervanadate reversed the inhibitory effect of C2 toxin on stimulated protein tyrosine phosphorylation indicating activation of phosphatases by C2 toxin. The data indicate that disassembly of the actin cytoskeleton by C2 toxin facilitates Fc epsilonRI-mediated signal-secretion coupling and suggest a role of the actin cytoskeleton in phosphatase regulation in RBL cells.


Maximising value from PFI contracts.
Against a backdrop where the Coalition Government has said more 'value' needs to be squeezed out of existing healthcare PFI projects, Karen Prosser, head of the health sector team at built asset consultancy, EC Harris, and Russell Gates, one of the company's partners on the same team, set out some of the key elements that NHS Trusts with operational PFI contracts should consider when undertaking a contract savings review.


Notch signaling in vascular smooth muscle cells is required to pattern the cerebral vasculature.
Stroke is the third leading cause of death and a significant contributor of morbidity in the United States. In humans, suboptimal cerebral collateral circulation within the circle of Willis (CW) predisposes to ischemia and stroke risk in the setting of occlusive carotid artery disease. Unique genes or developmental pathways responsible for proper CW formation are unknown. Herein we characterize a mouse model lacking Notch signaling in vascular smooth muscle cells (vSMCs), in which the animals are intolerant to reduced cerebral blood flow. Remarkably, unilateral carotid artery ligation results in profound neurological sequelae and death. After carotid ligation, perfusion of the ipsilateral cerebral hemisphere was markedly diminished, suggesting an anastomotic deficiency within the CW. High-resolution microcomputed tomographic (micro-CT) imaging revealed profound defects in cerebrovascular patterning, including interruption of the CW and anatomic deformity of the cerebral arteries. These data identify a vSMC-autonomous function for Notch signaling in patterning and collateral formation within the cerebral arterial circulation. The data further implicate genetic or functional deficiencies in Notch signaling in the pathogenesis of anatomic derangements underlying cerebrovascular accidents.


[A study of preoperative administration of 5'-DFUR in patients with colo-rectal cancer].
For the purpose to assess tumor-selectivity of doxifluridine (5'-DFUR), an anticancer drug of fluorinated pyrimidine class, the authors conducted a clinical-pharmacological study with the drug. The subjects in the present study, 26 patients with colorectal cancer received surgeries, were preoperatively administered 5'-DFUR orally at either doses of 800 or 1,200 mg/day for 3 days followed by oral administration of the drug at a dose of 400 mg 4-6 hrs before operation. The tissues were collected from specimens removed at operations and we measured both the activities of pyrimidine nucleoside phosphorylase (PyNPase), an activating enzyme for 5'-DFUR, and 5-FU concentrations in the tissue samples. The PyNPase activities showed higher levels in tumor tissues and regional lymph nodes than in normal tissues adjacent to the tumors. The 5-FU concentrations were: 102.7 ng/g in tumor tissues, 12.5 ng/g in normal tissues adjacent to the tumors, 97.6 ng/g in metastatic lymph nodes, and 7.3 ng/g in normal lymph nodes. In other words, the 5-FU concentrations were significantly (p < 0.01) higher in either of tumor tissues and metastatic lymph nodes than in the rests. The 5-FU concentrations in the sera were also extremely low. The results above clearly indicate that 5'-DFUR has a high selectivity for tumors.


Freud's three theories of love in the light of later developments.
Evidence is presented that Freud developed three very different theories on love. These theories were not integrated into a coherent theory. In subsequent developments each theory had its own history. The paper discusses the history of the controversy on the genital character, the relation between love and gender identity, between love and narcissism, the hierarchical structure of the capacity to love, and the relation between love and object loss. The impact of some concepts such as symbiosis and the rapprochmente subphase on the understanding of conflicts in loving is raised. While at present differences in emphasis make it difficult to build a coherent psychoanalytic theory of love, it is productive to bring divergent views in touch with each other. A unified theory of love based on psychoanalytic observations is suggested.


Immunogenic protein variations of Clostridium chauvoei cellular antigens associated with the culture growth phase.
The immunoprotective capacity of four Clostridium chauvoei strains at different growth stages is reported. In all the strains tested, the cells coming from the stationary phase were those with the highest immunoprotective capacity and, depending on the strain, this protective capacity diminished or even disappeared in other phases. Protein profiles were similar in all the strains and few proteins were differentially expressed during growth as shown by SDS-PAGE. For strain 17, a local strain, a clear relationship was observed between the diminution of immunogenicity and the total loss of protective capacity of sonicated cells at late stationary phase.


Implant Alignment and Patient Factors Affecting the Short-Term Patient-Reported Clinical Outcomes after Oxford Unicompartmental Knee Arthroplasty.
This study aims at clarifying implant alignment and other patient factors' influence on clinical outcomes, particularly on patient-reported outcomes (PRO), following Oxford unicompartmental knee arthroplasty (OUKA). A total of 142 patients after OUKA were divided into two groups according to the validated Japanese version of the knee injury and osteoarthritis outcome score (KOOS) for each subscale of pain, symptoms, and activities of daily living (ADL) at postoperative year 1 and 2: group 1 had ≥80 scores and group 2 had <80 scores. Postoperative clinical and radiographical findings were then compared among groups in each subscale. Using postoperative year 1 and 2 data, a multivariable logistic regression analysis was conducted to clarify factors for clinical outcomes' improvement. In the analysis of KOOS subscale of symptoms at postoperative year 1, gender distribution, preoperative body mass index, and postoperative maximum knee flexion angles differed significantly among groups. In the analysis for KOOS subscale of ADL, significant differences were observed in postoperative tibial component varus angles. Multivariable logistic regression analysis revealed that all of these parameters significantly related to ≥80 KOOS scores of each subscale at postoperative year 1. Additionally, at postoperative year 2, larger postoperative knee flexion angles and tibial component varus angles related to ≥80 KOOS subscale of symptoms and ADL were observed. In conclusion, tibial component's larger varus alignment was associated with the better PRO at both 1 and 2 years after OUKA. Importantly, larger postoperative knee flexion angle helped achieve good short-term PRO after OUKA.


Deficiencies in suicide training in primary care specialties: a survey of training directors.
A high percentage of suicide victims have seen a primary care physician in the months before committing suicide. Thus, primary care physicians may play an important role in suicide prevention. The authors mailed a survey to directors of training programs in family practice, internal medicine, and pediatrics, and 50.5% responded. Data obtained were analyzed with WebStat. Training directors reported deficiencies in training in suicide and depression. Notably, less than half of the internal medicine and pediatrics training directors who replied reported that teaching about suicide was adequate. The majority of them indicated a need for standardized curricular materials on suicide and depression. Experts could provide standardized curricula to primary care residencies in the recognition and management of suicide and depression. More robust training about these vital mental health concerns in primary care could reduce morbidity and mortality.


[Study on the Influence of Mineralizer on the Preparation of Calcium Aluminates Based on Infrared Spectroscopy].
In this study, effect of mineralizer on the structure and spectraproperties of calcium aluminates formation was extensively studied. Medium or low-grade bauxite and calcium carbonate were used as raw material and mineralizer CaF2 as additive. Calcium aluminates can be obtained after mixing fully, calcination and grinding. The prepared calcium aluminates can be directly used for the production of polyaluminiumchloride (PAC), polymeric aluminum sulfate, sodium aluminate and some other water treatment agents. The calcium aluminates preparation technology was optimized by investigating the mass ratio of raw materials (bauxiteand calcium carbonate) and mineralizer CaF2 dosage. The structure and spectra properties of bauxite and calcium aluminates were characterized by Fourier transform infrared(FTIR) spectroscopy analysis and the mineralization mechanism of the mineralizer was studied. FTIR spectra indicated that the addition of mineralizer promoted the decomposition and transformation of the diaspore, gibbsite and kaolinite, the decomposition of calcium carbonate, and more adequately reaction between bauxite and calcium carbonate. In addition, not only Ca in calcium carbonate and Si in bauxite were more readily reacted, but also Si-O, Si-O-Al and Al-Si bonds in the bauxite were more fractured which contributed to the release of Al in bauxite, and therefore, the dissolution rate of Al2O3 could be improved. The dissolution rate of Al2O3 can be promoted effectively when the mineralizer CaF2 was added in a mass ratio amount of 3%. And the mineralizer CaF2 cannot be fully functioned, when its dosage was in a mass percent of 1. 5%. Low-grade bauxite was easier to sinter for the preparation of calcium aluminates comparing with the highgrade one. The optimum material ratio for the preparation of calcium aluminates calcium at 1 250 °C was the mass ratio between bauxite and calcium carbonate of 1 : 0. 6 and mineralizer CaF2 mass ratio percent of 3%.


A novel L-neopentylglycine derivative as auxiliary for copper-catalyzed asymmetric Michael reactions.
L-Neopentylglycine diethylamide (4a). was prepared from the new unnatural amino acid L-neopentylglycine (1). The utilization of amide 4a as a chiral auxiliary in the copper(II)-catalyzed asymmetric Michael reaction was investigated in comparison with L-valine diethylamide (4b). Cyclic beta-oxocarboxylates 7 react with 4a and 4b to give the respective enaminoesters 8, which were converted with methyl vinyl ketone (9) in the presence of 10 mol% Cu(OAc)(2). H2O at room temperature in acetone to yield the optically active Michael addition products (R)-10a, b with high selectivity independent of the starting enamine. In the case of the seven-membered beta-oxocarboxylate 7c, however, the valine-derived enamine 8f led to higher enantioselectivity for product 10c. Despite the bulkiness of the neopentyl group, the isopropyl group with an alpha-branch has a better stereoinducing effect.


Phylogeography of marine mutualists: parallel patterns of genetic structure between obligate goby and shrimp partners.
The survival of many organisms depends on interspecific, mutualistic interactions. Hence, it is important to assess the phylogeography of multiple mutualistic species simultaneously to gain insight into how their metapopulations persist, spread and recover from disturbance. Consequently, we sequenced mitochondrial DNA (cytochrome b) from a gobiid fish (Ctenogobiops feroculus) and its mutualistic partner, an alpheid snapping shrimp (Alpheus djeddensis). These obligate mutualists are common in shallow coral reef lagoons, and we collected individuals from 11 Indo-Pacific islands that were likely exposed to different disturbance histories due to sea level fluctuations associated with glaciation events [geological studies indicate that eastern oceanic islands (Cook Islands, French Polynesia) were more disturbed than western continental plate islands that have deeper, more extensive lagoons (Okinawa, Fiji)]. Both phylogenetic and population genetic analyses indicated that gobies from Okinawa and Fiji were genetically distinct both from each other and those in the Cook Islands and French Polynesia. Shrimp, by contrast, grouped into two distinct genetic groups that showed no geographic structure. Restricting the analysis to one clade of shrimp, which contained two-thirds of the individuals, showed that these shrimp had very similar phylogeographical structure to the gobies. Mismatch distributions demonstrated that both gobies and shrimp experienced a recent, rapid population expansion into French Polynesia. Finally, nucleotide and haplotype diversities per sample location were significantly correlated between mutualists and tended to be higher on western continental than eastern oceanic islands. Our results suggest that these mutualists recovered from a major disturbance by colonizing South Pacific islands in synchrony.


A genomically modified Escherichia coli strain carrying an orthogonal E. coli histidyl-tRNA synthetase•tRNAHis pair.
Development of new aminoacyl-tRNA synthetase (aaRS)•tRNA pairs is central for incorporation of novel non-canonical amino acids (ncAAs) into proteins via genetic code expansion (GCE). The Escherichia coli and Caulobacter crescentus histidyl-tRNA synthetases (HisRS) evolved divergent mechanisms of tRNAHis recognition that prevent their cross-reactivity. Although the E. coli HisRS•tRNAHis pair is a good candidate for GCE, its use in C. crescentus is limited by the lack of established genetic selection methods and by the low transformation efficiency of C. crescentus. E. coli was genetically engineered to use a C. crescentus HisRS•tRNAHis pair. Super-folder green fluorescent protein (sfGFP) and chloramphenicol acetyltransferase (CAT) were used as reporters for read-through assays. A library of 313 ncAAs coupled with the sfGFP reporter system was employed to investigate the specificity of E. coli HisRS in vivo. A genomically modified E. coli strain (named MEOV1) was created. MEVO1 requires an active C. crescentus HisRS•tRNAHis pair for growth, and displays a similar doubling time as the parental E. coli strain. sfGFP- and CAT-based assays showed that the E. coli HisRS•tRNAHis pair is orthogonal in MEOV1 cells. A mutation in the anticodon loop of E. coli tRNAHisCUA elevated its suppression efficiency by 2-fold. The C. crescentus HisRS•tRNAHis pair functionally complements an E. coli ΔhisS strain. The E. coli HisRS•tRNAHis is orthogonal in MEOV1 cells. E. coli tRNAHisCUA is an efficient amber suppressor in MEOV1. We developed a platform that allows protein engineering of E. coli HisRS that should facilitate GCE in E. coli. This article is part of a Special Issue entitled "Biochemistry of Synthetic Biology - Recent Developments" Guest Editor: Dr. Ilka Heinemann and Dr. Patrick O'Donoghue.


Rifaximin Use, Adherence and Persistence in Patients with Hepatic Encephalopathy: A Real-World Study in the South of Italy.
Real-world data on the therapeutic management of hepatic encephalopathy (HE) patients are limited. The aim of this study was to evaluate the HE medications prescribed in an Italian cohort of HE patients post-discharge and to assess the real-world rifaximin adherence and persistence over 1 year. An observation retrospective study was conducted using data retrieved from outpatient pharmaceutical databases and hospital discharge records of the Campania region. For all subjects hospitalized for HE during 2019 (cohort 1), the HE medications prescribed within 60 days after discharge were evaluated. Adherence (proportion of days covered, PDC) and persistence were estimated for rifaximin 550 mg incident users over 1 year (cohort 2). Patients with PDC ≥80% were considered adherents. Persistence was defined as the period of time from the first rifaximin prescription to the date of discontinuation. Discontinuation was assessed using the permissible gap method. In cohort 1, 544 patients were identified; 58.5% received rifaximin while 15.6% only received non-absorbable disaccharides and 25.9% did not receive any HE medications. In cohort 2, 650 users were selected; only 54.5% were adherents and 35% were persistent users at 1 year. This real-world study highlights that quality improvement in therapeutic management is needed to potentially improve the outcomes of HE patients.


Tonsillar B cells do not express PSGL-1, but a significant fraction displays the cutaneous lymphocyte antigen and exhibits effective E- and P-selectin ligand activity.
Skin-homing T cells are defined by the expression of the cutaneous lymphocyte-associated antigen (CLA) which enables the cells to selectively bind to vascular endothelial E-selectin close to sites of cutaneous inflammation, an initial step in the effective extravasation from blood into the inflamed tissue. Essentially all CLA on T cells decorates the backbone of the P-selectin glycoprotein ligand-1 (PSGL-1). In this study we show that human peripheral blood B cells (PBBC) and tonsillar B cells (TBC) do not display PSGL-1 in fluorescence-activated cell sorter analysis using different murine monoclonal antibodies and polyclonal rabbit anti-PSGL-1 antiserum. A significant population of TBC, however, expresses a HECA-452-reactive epitope. These cells represent nonactivated IgM(+)/IgG(-) mature B lymphocytes. Up to 50% of the TBC in a given preparation strongly bind to E- and up to 79% to P-selectin. The shear stress resistance in a parallel-plate flow chamber system was high. Neuraminidase treatment of TBC totally and O-sialoglycoprotein endopeptidase partially diminished HECA-452 reactivity and reduced E- but not P-selectin ligand activities. Mocarhagin had no effect in the assays. The data suggest a different ligand for P-selectin and a distinct glycoprotein carrier for the E-selectin ligand as compared to T cells or other leukocytes. Adhesion to P-selectin, however, still required sulfation of the ligand for function. Western blots of TBC cell lysates detected a >240-kD HECA-452-reactive material that was resistant to reducing conditions. Anti-PSGL-1 did not reveal immunoreactive material in these cell lysates. B cell activation did neither significantly change HECA positivity nor induce PSGL-1 expression. Cultured, activated TBC, however, maintained expression of the integrin alpha4beta7. Human peripheral blood B cells had similar cell surface characteristics to TBC. Our observations suggest that several adhesion molecules may be involved in B cell homing which include CLA, the P-selectin ligand, and structures such as alpha4beta7.


Alpha-interferon therapy for chronic hepatitis C may induce acute allograft rejection in kidney transplant patients with failed allografts.
In hepatitis C virus (HCV) positive kidney transplant (KT) patients, the use of alpha-interferon (alphaIFN) is contraindicated due to the risk of acute rejection (AR). Conversely, if these HCV(+) KT patients lose their allograft, re-transplantation might be contemplated provided alphaIFN therapy has been attempted. Between 01/01/1989 and 31/12/1994, 261 kidney transplantations were performed; of these 174 were HCV(-) (group I) and 87 were HCV(+) (group II). At last follow-up (2006), in group I, the number of patients with a functioning graft, the number of patients who died with a functioning graft, and the number of patients who lost their graft before or after month (M) 12 were 92 (52.8%), 14 (8%), 20 (11.5%) and 48 (27.7%), respectively. In group II, the corresponding figures were 22 (25.3%; P < 0.0001), 8 (9.1%; ns), 9 (10.3%; ns) and 48 (55.3%; P < 0.0001). In group I, 19 of 48 (39.5%) patients with failed allografts after M12 underwent transplantectomy (TX) compared to 14 of 48 (29%; ns) in group II. In group II, 11 of 48 (23%) patients were offered alphaIFN therapy after their allograft failed: of these, four (36.3%) developed AR during alphaIFN therapy leading to TX. Histology, in addition to chronic allograft lesions, showed acute cellular and vascular lesions. In patients who were not offered alphaIFN therapy, TX was performed less frequently, i.e. in only six cases (16.2%). We conclude that even alphaIFN-treated KT patients with a failed allograft can experience acute allograft rejection that requires transplantectomy during therapy.


Nano magnetic solid phase extraction for preconcentration of lead ions in environmental samples by a newly synthesized reagent.
In this study, magnetite nanoparticles with particle size lower than 47 nm were synthesized and were applied for preconcentration of Pb2+ ions from aqueous solutions. To preconcentrate the Pb2+ ions, the surface of the synthesized nano particles was modified with sodium dodecyl sulfate (SDS) as an anionic surfactant. A new chelating agent (2-((E)-2-amino-4,5-dinitrophenylimino)methyl)phenol) was synthesized and used to form a very stable complex with Pb2+ ions. The lead ions formed complexes and were quantitatively extracted with SDS-coated magnetite nanoparticles. After magnetic separation of adsorbent, the adsorbent was eluted with 0.5% (v/v) HC1 in dimethyl sulfoxide (DMSO) prior to analysis by flame atomic absorption spectrometry (FAAS). Orthogonal array design (OAD) was used to study and optimize the different experimental parameters. Under the optimum conditions, enhancement factor up to 63.5 was achieved for extraction from only 10 mL of sample solution and the relative standard deviation (RSD %) of the method was lower than 2.8%. The obtained calibration curve was linear in the range of 1-300 pg L-' with reasonable linearity (r2 > 0.998). The limit of detection (LOD) based on S/N = 3 was 0.04 microg L(-1) for 10 mL sample volumes. Finally, applicability of the proposed method was successfully confirmed by preconcentration and determination of trace amounts of lead ions in environmental samples and satisfactory results were obtained.


Using antifibrinolytics in the peripartum period - concern for a hypercoagulable effect?
Although antifibrinolytic agents are used to prevent and treat hemorrhage, there are concerns about a potential increased risk for peripartum venous thromboembolism. We sought to determine the impact of tranexamic acid and ɛ-aminocaproic acid on in vitro clotting properties in pregnancy. Blood samples were obtained from healthy pregnant, obese, and preeclamptic pregnant women (n = 10 in each group) prior to delivery as well as from healthy non-pregnant controls (n = 10). Maximum clot firmness (MCF) and clotting time (CT) were measured using rotation thromboelastometry in the presence of tranexamic acid (3, 30, or 300 μg/mL) or ɛ-aminocaproic acid (30, 300, or 3000 μg/mL). ANOVA and regression analyses were performed. Mean whole blood MCF was significantly higher in healthy pregnant vs. non-pregnant women (66.5 vs. 57.5 mm, p < 0.001). Among healthy pregnant women, there was no significant difference between mean MCF (whole blood alone, and with increasing tranexamic acid doses = 66.5, 66.1, 66.4, 66.3 mm, respectively; p = 0.25) or mean CT (409, 412, 420, 424 sec; p = 0.30) after addition of tranexamic acid. Similar results were found using ɛ-aminocaproic acid. Preeclamptic women had a higher mean MCF after the addition of ɛ-aminocaproic acid and tranexamic acid (p = 0.05 and p = 0.04, respectively) compared to whole blood alone. Pregnancy is a hypercoagulable state, as reflected by an increased MCF compared to non-pregnant women. Addition of antifibrinolytic therapy in vitro does not appear to increase MCF or CT for non-pregnant, pregnant, and obese women. Whether antifibrinolytics are safe in preeclampsia may require further study.


Development of an extraocular retinal prosthesis: evaluation of stimulation parameters in the cat.
Electrical stimulation of the retina with extraocular electrodes is a new approach to developing a retinal prosthesis for blind patients. We have evaluated stimulus and electrode configurations for an extraocular retinal prosthesis (ERP). In anesthetized cats, ERP disc electrodes of 1 mm, 2 mm and 3 mm diameter were sutured to the sclera over the lateral globe. Electrically evoked potentials (EEPs) were recorded over the ipsilateral visual cortex, which resulted from the retinal stimulation of the ERP electrodes with a return electrode placed at the medial canthus. Square pulses, triangular pulses and the effects of dark adaptation and electrode size on the amplitude and thresholds for a cortical response were investigated. Square pulses were more effective than triangular pulses for stimulating the retina. Dark adaptation leads to a large increase in the threshold for retinal stimulation. There was no difference in the threshold for stimulation between electrodes of 1 mm and 3 mm diameter. Stimulation of the retina with extraocular electrodes elicits an EEP that is similar to that generated by retinal stimulation with intraocular electrodes. The use of square pulses is preferred to triangular pulses to minimize the peak current density at the electrode-tissue interface. As there is little difference in the threshold current for retinal stimulation with 1 mm or 3 mm electrodes, 3 mm electrodes are preferred as this will decrease the charge density at the active surface of the electrode.


Gastroesophageal reflux disease.
Chronic GERD is an unremitting, incurable disorder that recurs rapidly upon discontinuation of therapy. Primary complications of GERD include esophagitis, esophageal stricture and Barrett esophagus. Current therapy focuses on modifying risk factors, inhibiting the production of acid and enhancing esophageal gastric motility. In patients with uncomplicated heartburn, nondrug therapy should be the initial therapeutic approach, with patient education a major step in promoting lifestyle changes and improving the outcome. Maintenance therapy is central to the management of GERD. If symptoms persist to suggest complicated disease, further diagnostic tests (endoscopy) are indicated. H2 receptor antagonists usually resolve symptoms in 50% to 70% of patients, and PPIs in 74% to 96% of patients. Agents that improve esophageal motility, such as cisapride, may provide symptomatic relief of heartburn, but healing effects are inconsistent. In refractory disease, therapy is individualized to the patient, and may include combination therapy, more aggressive single line therapy or an appropriate surgical approach.


Pharmacokinetic interactions between theophylline and other medication (Part I).
Many drugs have been found to increase or decrease the clearance of theophylline, probably by interaction with one or more of the variants of the cytochrome P450 drug-metabolising system. Theophylline may be particularly susceptible to alteration of its clearance because of the particular form(s) of the P450 system involved, because its metabolism is saturable, and/or because 90% of its elimination is via metabolism. Its clearance has been found to be decreased (typically by around 25%, but often by far more) by erythromycin, troleandomycin (triacetyloleandomycin), roxithromycin, enoxacin, ciprofloxacin, pefloxacin, norfloxacin, ofloxacin, fluoroquinolone T-3262, pipemidic acid, cimetidine, etintidine, propranolol, verapamil, diltiazem, nifedipine, furosemide (frusemide), at least some anovulent agents, viloxazine, allopurinol, ticlopidine, idrocilamide, thiabendazole, disulfiram, influenza- and BCG-vaccination, interferon, and caffeine (half-life increase). In contrast, theophylline clearance (clearance/bioavailability) was found to be increased by isoprenaline (isoproterenol), terbutaline, some corticosteroids, phenytoin, phenobarbital, activated charcoal, felodipine moricizine, benzodiazepines and sulfinpyrazone - typically by about 25%, but sometimes by as much as 80% or more. For several of these concomitant medications, however, only some of the published studies can substantiate an influence, which may highlight the sensitivity of some interactions to particular experimental and/or clinical conditions, e.g. with terbutaline, erythromycin, ciprofloxacin, norfloxacin, ofloxacin, phenobarbital, cimetidine, verapamil, diltiazem, nifedipine, anovulents, allopurinol and influenza vaccination. Moreover, reports both of inhibition and of induction of theophylline clearance by each of rifampicin and isoniazid have appeared. Nevertheless, under investigation many medications have not been found to perceptibly influence theophylline disposition kinetics, e.g. ephedrine, orciprenaline (metaproterenol), prednisone, prednisolone, temelastine, terfenadine, mequitazine, picumast, repirinast, josamycin, midecamycin, miocamycin, spiramycin, amoxicillin, ampicillin, cefalexin, cefaclor, ceftibuten, cotrimoxazole (trimethoprim plus sulfamethoxazole), tetracycline, doxycycline, lomefloxacin, fluoroquinolones NY-198 and AM-833, nalidixic acid, lincomycin, metronidazole, certain antacids, ranitidine, roxatidine, pirenzepine, rioprostil, metoclopramide, metoprolol, atenolol, nadolol, medroxyprogesterone, dextropropoxyphene (propoxyphene), piroxicam, ozagrel, mebendazole and ascorbic acid.(ABSTRACT TRUNCATED AT 400 WORDS)


The Drosophila sex determination gene snf encodes a nuclear protein with sequence and functional similarity to the mammalian U1A snRNP protein.
Alternative splicing controls the expression of many genes, including the Drosophila sex determination gene Sex-lethal. Previous studies have suggested that snf plays a role in regulating Sex-lethal splicing. Here, we demonstrate that snf is an integral component of the machinery required for splice site recognition. We have cloned snf and found that it has sequence homology to the mammalian U1A and U2B" snRNP proteins. Moreover, we establish that snf encodes a Drosophila protein shown previously to have functional similarity to U1A. Finally, with the isolation and analysis of a null mutation, we demonstrate that snf is an essential gene. These studies provide the first demonstration, in a multicellular organism, that mutations in a U1 snRNP protein alter splicing in vivo.


Success of the state pain initiatives: moving pain management forward.
The purpose of this article is to describe the history of the State Cancer Pain Initiative (CPI) movement and current initiative activities that are directed at improving the management of cancer pain. The undertreatment of pain is a major public health problem in the United States. State CPIs strive to remove the barriers to effective cancer pain management through the provision of innovative professional, patient, and public educational programs, through institutional change projects, and through public advocacy. Collaboration with organizations that share common goals is important in sustaining and maximizing the impact of the movement. Professionals, patients, and families benefit from the work of the State Initiatives. By enhancing the knowledge and skills of clinicians, supporting institutional change, and teaching patients and families the importance of effective pain control, the Initiatives help to improve the quality of life for all cancer survivors. Models of intervention that respond to local needs are discussed, and legislative and advocacy efforts that affect care also are noted.


Correlation of dystrophin-glycoprotein complex and focal adhesion complex with myosin heavy chain isoforms in rat skeletal muscle.
The dystrophin-glycoprotein complex (DGC) and focal adhesion complex (FAC) are transmembrane structures in muscle fibres that link the intracellular cytoskeleton to the extracellular matrix. DGC and FAC proteins are abundant in slow-type muscles, indicating the structural reinforcement which play a pivotal role in continuous force output to maintain posture for long periods. The aim of the present study was to examine the expression of these structures across fast-type muscles containing different myosin heavy chain (MHC) isoform patterns which reflect the fatigue-resistant characteristics of skeletal muscle. We measured the expression of dystrophin and beta1 integrin (representative proteins of DGC and FAC respectively) in plantaris, extensor digitorum longus, tibialis anterior, red and white portions of gastrocnemius, superficial portion of vastus lateralis and diaphragm, in comparison with soleus (SOL) and cardiac muscle from rats. The expression of dystrophin and beta1 integrin correlated positively with the percentage of type I, IIa and IIx MHC isoforms and negatively with that of type IIb MHC isoform in fast-type skeletal muscles, and their expression was abundant in SOL and cardiac muscle. Our results support the idea that DGC and FAC are among the factors that explain the fatigue-resistant property not only of slow-type but also of fast-type skeletal muscles.


Clostridial Enterotoxemia and Coccidiosis in Weanling Cottontail Rabbits ( Sylvilagus audubonii, Sylvilagus floridanus, Sylvilagus nuttallii) from Colorado, USA.
Wild cottontail rabbits ( Sylvilagus spp.), especially young individuals, are one of the most frequent wildlife species presented for rehabilitation at wildlife rehabilitation centers. These species are challenging to rehabilitate, with gastrointestinal (GI) disease being a major cause of morbidity and mortality during the weaning stage. Two organisms, Clostridium spiroforme and Eimeria spp., are frequently associated with GI disease in young domestic rabbits ( Oryctolagus cuniculus) and can result in high rates of morbidity and mortality in this species. Here we present evidence that these two pathogens also play an important role in GI disease in young cottontail rabbits ( Sylvilagus audubonii, Sylvilagus floridanus, Sylvilagus nuttallii) undergoing rehabilitation.


Multiannual Assessment of Quality of Plantago major L. Seed Progeny from Kyshtym Radiation Accident Area: Weather-Dependent Effects.
The effects of low-dose radiation that are observed in plant populations in radioactively contaminated areas are variable. One of the reasons is the influence of fluctuating weather conditions and the interaction of radiation with weather factors. This article summarizes results of 12-year research on the viability and radioresistance of greater plantain (Plantago major L.) seed progeny growing in the East Ural Radioactive Trace (EURT) zone and in control (nonradioactive) areas, with consideration of weather conditions' variability. The EURT was formed by the Kyshtym accident, which occurred in 1957 at the Mayak Production Association. Absorbed dose rates of P. major parental plants in the pollution gradient were 14.5-165.9 μGy h-1, which correspond to a low-dose range. Seed progeny quality was evaluated as seed weight, the survival rate, and root length of 21-day seedlings. Interannual variability in the studied parameters was high, and their ranges overlapped between EURT groups of seeds and control groups in most cases. The number of significant correlations between the parameters of seed quality and weather conditions was higher in EURT groups than in control populations. In the control groups of seeds, 88.9% of correlations were negative, whereas in the EURT groups, 78.5% were positive.


Reduced Neuromuscular Performance in Amenorrheic Elite Endurance Athletes.
Secondary functional hypothalamic amenorrhea (SFHA) is common among female athletes, especially in weight-sensitive sports. The aim of this study was to investigate the link between SFHA and neuromuscular performance in elite endurance athletes. Sixteen eumenorrheic (EUM) and 14 SFHA athletes from national teams and competitive clubs participated. Methods included gynecological examination, body composition (dual-energy x-ray absorptiometry), resting metabolic rate and work efficiency, exercise capacity, knee muscular strength (KMS) and knee muscular endurance (KME), reaction time (RT), blood sampling performed on the third to fifth days of the menstrual cycle, and 7-d assessment of energy availability. SFHA athletes had lower estrogen (0.12 ± 0.03 vs 0.17 ± 0.09 nmol·L, P < 0.05), triiodothyronine (T3) (1.4 ± 0.2 vs 1.7 ± 0.3 nmol·L, P < 0.01), and blood glucose (3.8 ± 0.3 vs 4.4 ± 0.3 mmol·L, P < 0.001) but higher cortisol levels (564 ± 111 vs 400 ± 140 nmol·L, P < 0.05) compared with EUM athletes. SFHA had a lower body weight (55.0 ± 5.8 vs 60.6 ± 7.1 kg, P < 0.05), but no difference in exercise capacity between groups was found (56.4 ± 5.8 vs 54.0 ± 6.3 mL O2·min·kg). RT was 7% longer, and KMS and KME were 11% and 20% lower compared with EUM athletes. RT was negatively associated with glucose (r = -0.40, P < 0.05), T3 (r = -0.37, P < 0.05), and estrogen (r = -0.43, P < 0.05), but positively associated with cortisol (r = 0.38, P < 0.05). KMS and KME correlated with fat-free mass in the tested leg (FFMleg; r = 0.52, P < 0.001; r = 0.58, P < 0.001) but were negatively associated with cortisol (r = -0.42, P < 0.05; r = -0.59, P < 0.001). FFMleg explained the differences in KMS, while reproductive function and FFMleg independently explained the variability in KME. We found lower neuromuscular performance among SFHA compared with EUM athletes linked to a lower FFMleg, glucose, estrogen, T3, and elevated cortisol levels.


Molecular Weight Determinations.
A Beams magnetically suspended equilibrium ultracentrifuge was used to determine the molecular weight of sucrose, ribonuclease, and insulin. Both long- and short-column ultracentifuge cells were used. The longer cells gave greater precision, but required a longer time for equilibrium to occur.