Mathematical Model of Ultrasound Attenuation With Skull Thickness for Transcranial-Focused Ultrasound.
Transcranial-focused ultrasound (tFUS) has potential for both neuromodulation and neuroimaging. Due to the influence of head tissue, especially the skull, its attenuation is a key issue affecting precise focusing. The objective of the present study was to construct a mathematical model of ultrasound attenuation inclusive of skull thickness. First, combined with real skull phantom experiments and simulation experiments, tFUS attenuation of different head tissues was investigated. Furthermore, based on the system identification method, a mathematical model of ultrasound attenuation was constructed taking skull thickness into account. Finally, the performance of the mathematical model was tested, and its potential applications were investigated. For different head tissues, including scalp, skull, and brain tissue, the skull was found to be the biggest influencing factor for ultrasound attenuation, the attenuation caused by it being 4.70 times and 7.06 times that of attenuation caused by the brain and scalp, respectively. Consistent with the results of both the simulation and phantom experiments, the attenuation of the mathematical model increased as the skull thickness increased. The average error of the mathematical model was 1.87% in the phantom experiment. In addition, the experimental results show that the devised mathematical model is suitable for different initial pressures and different skulls with correlation coefficients higher than 0.99. Both simulation and phantom experiments validated the effectiveness of the proposed mathematical model. It can be concluded from this experiment that the proposed mathematical model can accurately calculate the tFUS attenuation and can significantly contribute to further research and application of tFUS.


Health system responsiveness for delivery care in Southern Thailand.
To assess the perception of women who gave birth in a hospital on health system responsiveness and their satisfaction. A cross-sectional study. Four district, one regional and one university hospitals in the Songkhla province, Southern Thailand. All women who delivered in the participating hospitals from November 2007 to December 2008. All women were interviewed at 24- or 48-h post partum by well-trained interviewers who worked independently outside the hospital. Eight components of health system responsiveness were measured. The factors associated with high rate of health responsiveness and its effect on the women's satisfaction was estimated by multiple logistic regression. A total of 2822 women were interviewed and their ages ranged from 12 to 48 years (mean ± SD: 27.6 ± 6.3). The components of health system responsiveness, which influenced the women's decision for delivering in a hospital were, in the order of importance, prompt attention, dignity, clear communication, autonomy, basic amenities, confidentiality, choice of provider and social support. The majority of women (>80%) gave high ratings for dignity, clear communication, prompt attention and autonomy. The type of hospital was a significant factor in all components of health responsiveness. Compared with women with universal coverage, women insured with the social security and civil servant medical benefit schemes gave higher ratings of dignity, confidentiality and choice of provider. Women's satisfaction for delivery care was significantly associated with high rates of all health responsiveness components, except choice of health providers. Prompt attention, dignity, clear communication and autonomy influenced women's decision to deliver in the hospital. High health system responsiveness is important for women's satisfaction.


Development of the Nursing Practice Readiness Scale for new graduate nurses: A methodological study.
To develop a scale to assess nursing practice readiness and verify its validity and reliability. Practice readiness refers to the attitude and characteristics of new employees that enable them to successfully adapt to the workplace. For new graduate nurses to receive on-the-job training appropriate to their individual competency level, a scale to assess nursing practice readiness is needed. A methodological study was conducted in two phases: (1) development of the scale through a literature review and interviews related to nursing practice readiness; (2) confirming construct validity, criterion-related validity and reliability of the developed scale. Data were collected from 430 new graduate nurses in South Korea. The validity and reliability of the scale were verified using maximum likelihood exploratory factor analysis with promax factor rotation, confirmatory factor analysis using maximum likelihood estimation, Pearson correlation, analysis of variance and Cronbach's alpha. The Nursing Practice Readiness Scale consisted of 35 items for five factors: clinical judgment and nursing performance, professional attitudes, patient-centeredness, self-regulation and collaborative interpersonal relationship. Confirmatory factor analysis results revealed an adequate model fit. Construct validity and criterion-related validity were verified and all factors achieved a Cronbach's α greater than 0.80. The developed scale can evaluate educational outcomes and determine the need for additional support, which can improve the workplace adaptation of new graduate nurses, patient safety and quality of care.


Stem cells in orthopedics: current concepts and possible future applications.
Stem cells are the cells that have the ability to divide for indefinite periods in culture and to give rise to specialized cells. Sources of these cells include embryo, umbilical cord and certain sites in adults such as the central nervous system [CNS] and bone marrow. Its use hold promise of wide spread applications particularly in areas of spinal cord injury, difficult non-unions, critical bone defects, spinal fusions, augmentation of ligament reconstructions, cartilage repair and degenerative disc disorders. This review article contains current information derived from Medline searches on the use in various orthopedic subspecialties. Some issues remain at the forefront of the controversy involving stem cell research - legislation, ethics and public opinion, cost and concentration methods. As is true with any new technology, the enthusiasm for this technology that has potential to influence virtually every orthopedic case management, must be balanced by subjecting it to stringent clinical and basic research investigations.


Stability analysis of the GAL regulatory network in Saccharomyces cerevisiae and Kluyveromyces lactis.
In the yeast Saccharomyces cerevisiae, interactions between galactose, Gal3p, Gal80p, and Gal4p determine the transcriptional status of the genes required for the galactose utilization. Increase in the cellular galactose concentration causes the galactose molecules to bind onto Gal3p which, via Gal80p, activates Gal4p, which induces the GAL3 and GAL80 gene transcription. Recently, a linear time-invariant multi-input multi-output (MIMO) model of this GAL regulatory network has been proposed; the inputs being galactose and Gal4p, and the outputs being the active Gal4p and galactose utilization. Unfortunately, this model assumes the cell culture to be homogeneous, although it is not so in practice. We overcome this drawback by including more biochemical reactions, and derive a quadratic ordinary differential equation (ODE) based model. We show that the model, referred to above, does not exhibit bistability. We establish sufficiency conditions for the domain of attraction of an equilibrium point of our ODE model for the special case of full-state feedback controller. We observe that the GAL regulatory system of Kluyveromyces lactis exhibits an aberration of monotone nonlinearity and apply the Rantzer multipliers to establish a class of stabilizing controllers for this system. Feedback in a GAL regulatory system can be used to enhance the cellular memory. We show that the system can be modeled as a quadratic nonlinear system for which the effect of feedback on the domain of attraction of the equilibrium point can be characterized using linear matrix inequality (LMI) conditions that are easily implementable in software. The benefit of this result is that a mathematically sound approach to the synthesis of full-state and partial-state feedback controllers to regulate the cellular memory is now possible, irrespective of the number of state-variables or parameters of interest.


Functional analysis of the human pro-opiomelanocortin promoter in the small cell lung carcinoma cell line DMS-79.
DMS-79 is a human cell line derived from a small cell lung carcinoma (SCLC), which expresses the pro-opiomelanocortin (POMC) gene. We took it as a model in which to study the mechanism of POMC gene expression in these tumors: precursor processing is altered and gene expression is essentially unresponsive to glucocorticoids. POMC gene structure appeared normal by Southern blot analysis, indicating that gene rearrangement was not responsible for its expression in DMS-79. Indeed, using transient expression of human POMC-luciferase fusion genes in DMS-79, we showed that (1) the normal human POMC promoter was functional in DMS-79, and (2) the same proximal promoter region (-417; + 21) produced the full transcriptional activity in DMS-79 and in the mouse pituitary cell line AtT-20. Progressive 5' deletion analysis revealed differences between AtT-20 and DMS-79: region (-611; -376) was active in AtT-20 and not in DMS-79, whereas region (-95; -161) was active in both cell lines and (-376; -417) was only active in DMS-79. The latter partially overlaps a motif homologous to the DE-2 rat element which confers the tissue-specific expression of POMC in AtT-20 cells; however, this motif had no effect in DMS-79. These data suggest that POMC gene transcription is achieved through a different set of transacting factors in DMS-79 and AtT-20. Altogether, our results provide evidence that DMS-79 is a valid model of tumors responsible for the ectopic ACTH syndrome and that the mechanism of POMC gene expression in these SCLC cells is different from that in pituitary cells.


Auditory hallucinations and posttraumatic stress disorder within schizophrenia and substance abuse.
There is a high prevalence of traumatic events within individuals diagnosed with schizophrenia and of auditory hallucinations within individuals diagnosed with posttraumatic stress disorder (PTSD). However, the relationship among the symptoms associated with these disorders remains poorly understood. We conducted a multidimensional assessment of auditory hallucinations within a sample diagnosed with schizophrenia and substance abuse, both with and without comorbid PTSD. The results suggest a rate of comorbid PTSD similar to those reported in other studies. Patients who have comorbid PTSD reported more distressing auditory hallucinations. However, the hallucinations were not more frequent or of longer duration. The need for a multidimensional assessment is supported. The results are discussed within current theoretical accounts of traumatic psychosis.


Chest pain associated with nutcracker esophagus: a preliminary study of the role of gastroesophageal reflux.
A review of our 402 motility records of patients undergoing evaluation of noncardiac chest pain identified 40 patients with the diagnosis of nutcracker esophagus. Gastroesophageal reflux was found in 13 of 20 patients (65%) who underwent pH studies, and endoscopy detected one patient with erosive esophagitis. Thus, at least 14 (35%) of our nutcracker esophagus patients had evidence of reflux. Twelve of these subjects agreed to enter an open-label therapeutic trial. After 8 wk of intensive antireflux treatment with high doses of ranitidine or omeprazole, repeat 24-h pH studies and endoscopy demonstrated normalization of pH parameters and healing of esophagitis in all patients. Ten (83%) patients obtained significant symptomatic improvement in frequency of pain episodes, number of days with pain, and pain severity. However, repeat manometry showed normalization of motor findings in only two (18%) patients. These observations warrant further placebo-controlled trials. Until more information is available, the results of this study suggest that gastroesophageal reflux should be excluded in patients with noncardiac chest pain and nutcracker esophagus before initiation of smooth muscle relaxant therapy.


Chevon production and quality of Kalahari Red goats fed increasing levels of hempseed cake substituted for soybean meal.
A 42-d study was conducted to evaluate the effects of replacing soybean meal with increasing levels of hempseed cake (HSC) in goat finishing diets on growth performance, carcass and chevon quality attributes. Thirty-five, 3-month-old Kalahari Red wethers (25 ± 1.5 kg initial body weight) were randomly allocated to one of five dietary treatments with seven animals per treatment. Wethers were fed maize-lucerne based finishing diets with inclusions of 0 (control), 25, 50, 75 and 100 g/kg DM of HSC replacing soybean meal as the main protein source. Diet had no effect (P > 0.05) on daily feed intake, average daily gain, final body weight and income-over-feed costs. Carcass and meat quality attributes were not influenced (P > 0.05) by HSC, except intramuscular fat, which increased linearly (P ≤ 0.05) with HSC inclusion levels. It was concluded that HSC could completely replace soybean meal in goat finishing diets without affecting chevon production and quality.


Pregnancy associated anemia and iron: a pilot study.
Iron deficiency occurs when the rate of loss of utilization exceeds its assimilation. Treatment is based on iron supplementation but due to side effects compliance to iron therapy is poor. A double blind comparative study was done using a novel time release preparation of ferrous sulphate (Code A) v.s. sustained release ferrous sulphate preparation (Code B) on 60 pregnant women in mid or late pregnancy with anaemia. The amount of ferrous sulphate in Code A was less than half of Code B. The patients were sequentially randomised as Code A or Code B recipient. The non compliance rate was 33%, and for both Code A & Code B. The mean improvement in hemoglobin after 4 weeks of therapy was 2.01 gm% for Code A and 2.3 gm% for Code B. Iron absorption as evidenced by improvement in S. Iron, TIBC and ferritin levels was better with Code A. The improvement in subjective symptoms of anaemia was better than average in Code B preparation. Code A group had comparatively more side effects both major and minor, this may have been the reason for a slightly higher drop out rate in this group. In conclusion the timed release preparation has a comparable haematological response and better absorption with significantly lower doses as compared to the sustained release preparation.


A test of the factor structure equivalence of the 50-item IPIP Five-factor model measure across gender and ethnic groups.
Personality is frequently assessed in research and applied settings, in part due to evidence that scores on measures of the Five-factor model (FFM) of personality show predictive validity for a variety of outcomes. Although researchers are increasingly using the International Personality Item Pool (IPIP; Goldberg, 1999; International Personality Item Pool, 2007b) FFM measures, investigations of the psychometric properties of these measures are unfortunately sparse. The purpose of this study was to examine the factor structure equivalence of the 50-item IPIP FFM measure across gender and ethnic groups (i.e., Whites, Latinos, Asian Americans) using multigroup confirmatory factor analysis. Results from a sample of 1,727 college students generally support the invariance of the factor structure across groups, although there was some evidence of differences across gender and ethnic groups for model parameters. We discuss these findings and their implications.


Principles Guiding the Square Bonding Motif Containing a Pair of Chalcogen Bonds between Chalcogenadiazoles.
The bonding motif adopted by a dimer of chalcogenadiazole molecules is characterized by a pair of equivalent Ch···N chalcogen bonds. Quantum calculations show that the interaction energy is substantial, varying between 4 kcal/mol for Ch = S and 17 kcal/mol for Te. The interaction is cooperative in that the total bond strength is greater than either chalcogen bond individually. Neither the addition of a phenyl ring nor the addition of a pair of cyano substituents to the diazole ring has much influence on this binding. Removal of one N from the diazole weakens the binding, and addition of two nitrogens has little effect. The largest perturbation arises with three N atoms in each ring, for which the binding energy increases by some 25%. The ring size plays a minor role in most cases, although a near doubling of bond strength occurs if there are two N atoms present on a four-membered ring.


Enhanced Efflux Activity Facilitates Drug Tolerance in Dormant Bacterial Cells.
Natural variations in gene expression provide a mechanism for multiple phenotypes to arise in an isogenic bacterial population. In particular, a sub-group termed persisters show high tolerance to antibiotics. Previously, their formation has been attributed to cell dormancy. Here we demonstrate that bacterial persisters, under β-lactam antibiotic treatment, show less cytoplasmic drug accumulation as a result of enhanced efflux activity. Consistently, a number of multi-drug efflux genes, particularly the central component TolC, show higher expression in persisters. Time-lapse imaging and mutagenesis studies further establish a positive correlation between tolC expression and bacterial persistence. The key role of efflux systems, among multiple biological pathways involved in persister formation, indicates that persisters implement a positive defense against antibiotics prior to a passive defense via dormancy. Finally, efflux inhibitors and antibiotics together effectively attenuate persister formation, suggesting a combination strategy to target drug tolerance.


Liquid crystalline phases of the amphiphilic ionic liquid N-hexadecyl-N-methylpyrrolidinium bromide formed in the ionic liquid ethylammonium nitrate and in water.
The phase behavior of a surfactant-like ionic liquid, N-hexadecyl-N-methylpyrrolidinium bromide (C(16)MPB), was studied in both water and a room temperature ionic liquid, ethylammonium nitrate (EAN). Polarized optical microscopy (POM) and small-angle X-ray scattering (SAXS) measurements were employed to investigate the phase behavior of the two systems and to determine which lyotropic liquid crystalline (LC) phases were formed. With increasing C(16)MPB concentration, an isotropic solution phase, a hexagonal (H(1)) phase, and a cubic phase (V(2)) are all present in either EAN or H(2)O. The structural parameters of the H(1) phase were calculated from SAXS patterns, which show the structural changes as a function of the amount of C(16)MPB. The rheological results reveal that the H(1) phase constructed by C(16)MPB in EAN displays a typical Maxwell behavior, whereas the H(1) phase formed by C(16)MPB in water shows a gel-like behavior, unlike traditional cationic surfactants. POM and differential scanning calorimetry (DSC) results demonstrate that the lyotropic LC phase in EAN has a higher thermal stability than that formed in H(2)O, which may be important to extend the applications of the LC phase.


Chloromethane-induced genes define a third C1 utilization pathway in Methylobacterium chloromethanicum CM4.
Methylobacterium chloromethanicum CM4 is an aerobic alpha-proteobacterium capable of growth with chloromethane as the sole carbon and energy source. Two proteins, CmuA and CmuB, were previously purified and shown to catalyze the dehalogenation of chloromethane and the vitamin B12-mediated transfer of the methyl group of chloromethane to tetrahydrofolate. Three genes located near cmuA and cmuB, designated metF, folD and purU and encoding homologs of methylene tetrahydrofolate (methylene-H4folate) reductase, methylene-H4folate dehydrogenase-methenyl-H4folate cyclohydrolase and formyl-H4folate hydrolase, respectively, suggested the existence of a chloromethane-specific oxidation pathway from methyl-tetrahydrofolate to formate in strain CM4. Hybridization and PCR analysis indicated that these genes were absent in Methylobacterium extorquens AM1, which is unable to grow with chloromethane. Studies with transcriptional xylE fusions demonstrated the chloromethane-dependent expression of these genes. Transcriptional start sites were mapped by primer extension and allowed to define three transcriptional units, each likely comprising several genes, that were specifically expressed during growth of strain CM4 with chloromethane. The DNA sequences of the deduced promoters display a high degree of sequence conservation but differ from the Methylobacterium promoters described thus far. As shown previously for purU, inactivation of the metF gene resulted in a CM4 mutant unable to grow with chloromethane. Methylene-H4folate reductase activity was detected in a cell extract of strain CM4 only in the presence of chloromethane but not in the metF mutant. Taken together, these data provide evidence that M. chloromethanicum CM4 requires a specific set of tetrahydrofolate-dependent enzymes for growth with chloromethane.


RAGE expression in tumor-associated macrophages promotes angiogenesis in glioma.
Interaction of RAGE (the receptor for advanced glycation endproducts) with its ligands can promote tumor progression, invasion, and angiogenesis. Although blocking RAGE signaling has been proposed as a potential anticancer strategy, functional contributions of RAGE expression in the tumor microenvironment (TME) have not been investigated in detail. Here, we evaluated the effect of genetic depletion of RAGE in TME on the growth of gliomas. In both invasive and noninvasive glioma models, animal survival was prolonged in RAGE knockout (Ager(-/-)) mice. However, the improvement in survival in Ager(-/-) mice was not due to changes in tumor growth rate but rather to a reduction in tumor-associated inflammation. Furthermore, RAGE ablation in the TME abrogated angiogenesis by downregulating the expression of proangiogenic factors, which prevented normal vessel formation, thereby generating a leaky vasculature. These alterations were most prominent in noninvasive gliomas, in which the expression of VEGF and proinflammatory cytokines were also lower in tumor-associated macrophages (TAM) in Ager(-/-) mice. Interestingly, reconstitution of Ager(-/-) TAM with wild-type microglia or macrophages normalized tumor vascularity. Our results establish that RAGE signaling in glioma-associated microglia and TAM drives angiogenesis, underscoring the complex role of RAGE and its ligands in gliomagenesis.


Leading limb preference during brachiation in the gibbon family member, Hylobates syndactylus (siamangs): a study of the effects of singing on lateralisation.
Individual-level lateralisations are common among vertebrates, however population-level preferences are usually reserved for specific tasks. In humans, handedness is thought to be related to the hemispheric processing of specific aspects of language, including speech. Although nonhuman primates do not possess speech, gibbons are known to produce elaborate vocal displays often referred to as song. To investigate the evolutionary effects of singing on hand preference, this study examined leading limb preference during brachiation in the large-bodied, melodious gibbon, Hylobates syndactylus (siamangs). A total of 13 male and 12 female siamangs were observed in captive and semi-captive settings for leading limb preference during vocal and nonvocal behaviour. No significant results were found for the effect of vocalisation at the group level, although individual-level data indicated a trend towards a right shift in the vocal condition, especially for females, where 8 out of 12 demonstrated a greater reliance on a right leading limb during the vocal versus nonvocal conditions. Males were not found to possess significant preferences in either the vocal or nonvocal conditions alone, however when condition data were combined, the number of individual males lateralised was significantly greater than predicted by chance. These findings support individual, but not population-level, effects for vocalisation on leading limb preference in siamangs, and emphasise the importance of testing for sex difference in handedness and lateralisation research.


Increased risk for respiratory distress among white, male, late preterm and term infants.
To determine whether race/ethnicity and sex independently increase risk of respiratory distress syndrome (RDS) in late preterm and term infants. Using a cohort design, we studied the risk of RDS associated with race/ethnicity and sex in infants with gestational age (GA) 34 to 42 weeks born between 1 January 2000 and 31 December 2009 (n=286 454) within 12 hospitals in the Northern California Kaiser Permanente Medical Care Program. Male sex (adjusted odds ratio (aOR) 1.68; 95% confidence interval 1.45 to 1.93) and White race/ethnicity (vs Asians (aOR 0.57; 95% confidence interval 0.47 to 0.70), Blacks (aOR 0.66; 95% confidence interval 0.50 to 0.87), and Hispanics (aOR 0.76; 95% confidence interval 0.64 to 0.90)) independently increase risk for RDS regardless of GA. A GA <39 weeks, operative delivery, maternal diabetes, and chorioamnionitis also increased RDS risk in this cohort. Male sex and White race/ethnicity independently increase risk for RDS in late preterm and term infants. Timing of elective delivery should acknowledge these risks.


Regulatory T cell subsets in bullous pemphigoid and dipeptidyl peptidase-4 inhibitor-associated bullous pemphigoid.
Regulatory T (Treg) cells play an essential role in peripheral immune tolerance. Bullous pemphigoid (BP) is the most common blistering disease and is caused by autoantibodies to two BP antigens: type XVII collagen and BP230. Recently, we reported that Treg cell dysfunction may cause the production of autoantibodies to BP antigens. Several studies have suggested an association between Treg cells and BP pathogenesis. However, Treg cells are heterogeneous in humans, leading to inconsistent results in previous studies. To assess functional Treg subsets in BP. We examined three distinct Treg subsets in conventional BP (cBP) patients before versus after systemic corticosteroid treatment, dipeptidyl peptidase-4 inhibitor-associated BP (DPP-4i-BP) patients, younger controls and older controls. We found that total Treg cells and all Treg cell subsets were increased in cBP patients before treatment and decreased by systemic corticosteroid treatment. In contrast, neither total Treg cells nor all Treg subsets were increased in DPP-4i-BP. Notably, CD45RA- Foxp3hi effector Treg cells positively correlated with disease severity in cBP, whereas CD45RA+Foxp3lo naïve Treg cells positively correlated with the disease severity in DPP-4i-BP. These findings suggest that Treg cells are differently involved in the pathogeneses of cBP and DPP-4i-BP.


Biochemical and x-ray diffraction analysis of concanavalin B crystals from jack bean.
Parallel biochemical and crystallographic studies have been carried out on the protein concanavalin B from Jack Bean (Canavalis ensiformis). The studies show the protein to be a monomer of 33,000 daltons with unexceptional amino acid composition and no covalently bound carbohydrate. The molecule contains a single, firmly bound zinc ion and we present evidence that the protein specifically binds nucleotide coenzymes including at least NADPH and flavin mononucleotide. An electron density map of the concanavalin B crystals (space group P6(1), a = b = 80.9 A, c = 102.2 A) has been calculated from phases based on six isomorphous heavy atom derivatives. A description of the structure of the protein based on the 5.0 A resolution x-ray diffraction study is provided. The structure appears to contain a substantial amount of alpha-helix as well as an extensive beta-sheet and these are organized into two domains of unequal size. The position of the bound zinc is indicated by anomalous difference Fouriers and the nucleotide binding site by conventional difference Fourier maps.


Obese patients show a depressed cytokine profile following severe blunt injury.
We hypothesized that severely injured obese patients would display increased concentrations of proinflammatory cytokines when compared with patients of normal body mass index (BMI) and that this would be associated with multiple organ failure (MOF). This was a retrospective review of prospectively collected data in the "Inflammation and the Host Response to Injury" trauma-related database. Data were collected prospectively from US level I trauma centers. The subjects were severely injured adult blunt trauma patients. Cytokine concentrations obtained within 12 h of injury and on days 1 and 4 were compared between subjects on the basis of BMI (normal, 18.5-24.9 kg/m, and obese, ≥30 kg/m). Demographic measures, injury severity, cytokine concentrations, and outcome measures were compared between groups. Seventy-four adult blunt trauma victims were evaluated. Relative to patients of normal BMI (n = 34), obese patients (n = 40) demonstrated an overall depressed cytokine response to severe injury, with significantly lower concentrations of several cytokines. Obese patients showed greater incidences of nosocomial infection (60 vs. 45%, not statistically significant) and MOF (63% vs. 44%, not statistically significant) and a later onset of maximum MOF score (5 vs. 3 days, P < 0.04) when compared with those of normal BMI. Despite prior reports suggesting a proinflammatory cytokine profile in obese individuals, obese patients sustaining severe injury show a depressed early cytokine response when compared with patients of normal BMI. This may confer increased susceptibility to nosocomial infection and later MOF. Further study of immune dysfunction in the postinjury obese patient should assess the possibility of early immune suppression.


[Effects of 3'-azido-deoxythymidine on telomerase activity and protein expression of hepatocarcinoma cell line SMMC-7721].
The mechanism of inhibiting telomerase activity by telomerase inhibitors is very complex, and involves common actions of many proteins. This study was to investigate the effects of 3'-azido-deoxythymidine (AZT) on telomerase activity and protein expression of hepatocarcinoma cell line SMMC-7721, and explore possible mechanism of inhibiting telomerase activity in SMMC-7721 cells by AZT. Optimized concentration and treatment time of AZT were detected by MTT assay. After treatment with AZT, the telomerase activity in SMMC-7721 cells was detected by real-time fluorescent quantitative TRAP (FQ-TRAP) assay; the apoptosis of SMMC-7721 cells was detected by TUNEL assay and flow cytometry (FCM); the changes of specific proteins were monitored by raman spectra assay and surface-enhanced laser desorption time of flight-mass spectrum (SELDI-TOF-MS) with protein chip assay. The optimized treatment time of AZT was 48 h, and the optimized concentration of AZT was 20 mmol/L. When treated with 20 mmol/L AZT for 48 h, the telomerase activity in SMMC-7721 cells was inhibited by 53.85% of control (P<0.001); apoptotic morphologic changes of SMMC-7721 cells were observed; the apoptosis rate of SMMC-7721 cells was 13.5%; 7 spectral peak of proteins were changed; 24 proteins were up-regulated and 8 were down-regulated in SMMC-7721 cells. All of the 32 distinct proteins were small molecular proteins. AZT can inhibit the telomerase activity and induce apoptosis of SMMC-7721 cells, which is related to specific small molecular proteins.


Incidence of multiple sclerosis in Møre and Romsdal, Norway from 1950 to 1991. An age-period-cohort analysis.
We have examined the time trends in multiple sclerosis incidence over the past four decades from 1950 to 1991 in Møre and Romsdal County, Norway. Incidence rates by year of birth, year of onset and year of diagnosis according to sex, age and clinical course were studied. All patients with multiple sclerosis diagnosed by August 1992 were included, giving a total of 419 patients (171 men, 248 women) with onset of multiple sclerosis in the period 1950-91. Mean age at onset was 33.2 years, and mean age at diagnosis was 39.2 years. The incidence rate by year of onset increased from 2.87 per 100 000 in the period 1950-54 (men 3.06, women 2.67) to 5.57 per 100 000 in 1985-91 (men 3.75, women 7.94) (P<0.001). The incidence was particularly high in the period from 1975 to 1985. Major period- or cohort-effects in incidence, apart from the general increasing trend, could not be identified by an age-period-cohort analysis.


Imidazodiazepinediones: a new class of adenosine receptor antagonists.
A series of imidazo[4,5-e][1-4]diazepine-5,8-diones were synthesized from hypoxanthines. Certain of these cyclic homologues of caffeine, theophylline, theobromine, 3-isobutyl-1-methylxanthine, and enprofylline were inhibitors of binding of adenosine analogues to rat brain A1 and A2 adenosine receptors and were antagonists of A2 adenosine receptors stimulatory to adenylate cyclase in rat PC12 cell membranes. Activity at adenosine receptors was lower than the corresponding xanthines, perhaps because imidazodiazepinediones contain a boat-shaped seven-membered ring rather than the planar heteroaryl ring system of the xanthines. The imidazodiazepinediones had low affinity for brain benzodiazepine sites.


Outcome of Listeria monocytogenes prosthetic valve endocarditis: as bad as it looks?
In 1998 we presented 1 successfully treated case of Listeria monocytogenes prosthetic valve endocarditis and made a review of all the cases that had been published to date. We carry out an up-to-date review through Pub-Med of every case of Listeria monocytogenes prosthetic valve endocarditis; mortality rate is calculated and data from several clinical and therapeutical variables are collected; Fisher's exact test is used to identify those variables significantly associated with mortality. Four out of 23 patients died in hospital (17%); among all the variables included, only peripheral embolism (p=0.024), onset on a mechanical prosthesis (p=0.035) and having used only 1 antibiotic instead of a combination of drugs (p=0.026) were independently associated with mortality. Although the overall number of cases is too low to draw definite conclusions (n=23), mortality rate is lower than previously believed. Some variables that have traditionally been associated with a poor prognosis for endocarditides are not for the case of Listeria monocytogenes on valvular prostheses. It seems prudent to treat affected patients with a combination of ampicillin -- or vancomycin, if there is a history of beta-lactam allergy and ampicillin desensitization is not possible -- plus an aminoglycoside.


Cytokine changes after a marathon race.
The influence of carbohydrate (1 l/h of a 6% carbohydrate beverage), gender, and age on pro- and anti-inflammatory plasma cytokine and hormone changes was studied in 98 runners for 1.5 h after two competitive marathon races. The marathoner runners were randomly assigned to carbohydrate (C, n = 48) and placebo (P, n = 50) groups, with beverages administered during the races in a double-blind fashion using color codes. Plasma glucose was higher and cortisol was lower in the C than in the P group after the race (P < 0.001). For all subjects combined, plasma levels of interleukin (IL)-10, IL-1 receptor antagonist (IL-1ra), IL-6, and IL-8 rose significantly immediately after the race and remained above prerace levels 1.5 h later. The pattern of change in all cytokines did not differ significantly between the 12 women and 86 men in the study and the 23 subjects > or =50 yr of age and the 75 subjects <50 yr of age. The pattern of change in IL-10, IL-1ra, and IL-8, but not IL-6, differed significantly between the C and the P group, with higher postrace values measured for IL-10 (109% higher) and IL-1ra (212%) in the P group and for IL-8 (42%) in the C group. In conclusion, plasma levels of IL-10, IL-1ra, IL-6, and IL-8 rose strongly in runners after a competitive marathon, and this was not influenced by age or gender. Carbohydrate ingestion, however, had a major effect in attenuating increases in cortisol and two anti-inflammatory cytokines, IL-10 and IL-1ra.


ORSAT and modifications of SEFT and APT.
Up to now, three subspectra are usually needed for the complete assignment of all signals in 13C spectrum: two DEPT spectra (phi = 90 degrees and phi = 135 degrees) and a proton decoupled spectrum. In this paper, we present a method in which a complete assignment becomes possible with merely two spectra. For this purpose, a new pulse sequence (ORSAT) has been elaborated by using off-resonance irradiation. The method described here is a further development of SEFT and APT. The second required spectrum is a DEPT (phi = 135 degrees). Signal assignment of cholesteryl acetate is demonstrated as an example. 13C routine spectroscopy can be significantly accelerated by applying this method.


Striving for Gerotranscendence: Preparing Nursing Students to Care for the Aging Population.
The population of older adults in America is increasing rapidly, but many prelicensure nursing students do not understand the unique needs of these patients. Educational methods are needed that foster a mindset of research and inquiry to help older adults thrive. A three-pronged educational hybrid was designed to help students create meaningful links between geriatric nursing theory in the classroom and application in the clinical setting. Test scores increased for 45% of students after the intervention, and 89% reported a greater understanding of evidence-based geriatric care. Students demonstrated increased confidence in discussing geriatric care and sharing theoretical knowledge with peers. With guided application of nursing theory in the clinical setting, students can use evidence-based care and develop nursing practice that helps older adults thrive. This prepares students to meet the demands of today's health care environment and develop practice standards that will follow them throughout their careers. [J Nurs Educ. 2020;59(9):536-538.].


Autism Spectrum Disorders and Metabolic Complications of Obesity.
To assess for an increased risk of obesity, type 2 diabetes mellitus, hypertension, hyperlipidemia, and nonalcoholic fatty liver disease/nonalcoholic steatohepatitis in children with autism spectrum disorders (ASD). Additionally, to determine the rates of prescribed treatment for obesity-related metabolic disorders and to determine whether treatment with psychotropic medications is associated with the development of obesity for children with ASD. A retrospective 1:5 case-control study was performed by use of the Military Health System database from October 2000 to September 2013. For children with ASD and matched controls, International Classification of Diseases, Ninth Revision, Clinical Modification diagnostic codes for obesity, type 2 diabetes mellitus, hypertension, hyperlipidemia, nonalcoholic fatty liver disease/nonalcoholic steatohepatitis, and prescriptions were obtained. Conditional logistic regression determined ORs and 95% CIs. A total of 48 762 individuals with ASD and 243 810 matched controls were identified. Children with ASD had significantly greater odds of having obesity (OR 1.85; 95% CI 1.78-1.92), having obesity-related disorders, and being prescribed a medication when they had these diseases. In children with ASD, mood stabilizers, antipsychotics, antiepileptic drugs, and selective serotonin reuptake inhibitors were associated with obesity. Children with ASD have an increased risk of obesity and obesity-related metabolic disorders. They are more likely to be prescribed medications to treat these complications, suggesting they may have more severe disease. There is a significant association between the use of some psychotropic categories and a diagnosis of obesity, suggesting that obesity in children with ASD may be partially iatrogenic.


Induction of electric fields due to gradient switching: a numerical approach.
Since the first observations of peripheral nerve stimulation in MRI, it has been clear that the underlying mechanism is the activation of the nervous system by induced electric fields. However, compared to experimental investigations little work has been done on calculating these electric fields with adequate accuracy. In this article a numerical analysis of the electric fields induced by a complete whole body gradient system is presented. The calculations were carried out on three human body models of different complexities. The numerical results correlate better to the experimental observations with a body model that resembles the human body. Applying a model with inhomogeneous conductivity, numerical stability was not reached. The results were compared to the limits given in the upcoming IEC 60601-2-33 standard. The comparison shows that the derived peak electric fields depend substantially on the body model used, which dictates that limits have to refer to a body model that is exactly defined.


Patient-individual local SAR determination: in vivo measurements and numerical validation.
Tissue heating during magnetic resonance measurements is a potential hazard at high-field MRI, and particularly, in the framework of parallel radiofrequency transmission. The heating is directly related to the radiofrequency energy absorbed during an magnetic resonance examination, that is, the specific absorption rate (SAR). SAR is a pivotal parameter in MRI safety regulations, requiring reliable estimation methods. Currently used methods are usually based on models which are neither patient-specific nor taken into account patient position and posture, which typically leads to the need for large safety margins. In this work, a novel approach is presented, which measures local SAR in a patient-specific manner. Using a specific formulation of Maxwell's equations, the local SAR is estimated via postprocessing of the complex transmit sensitivity of the radiofrequency antenna involved. The approximations involved in the proposed method are investigated. The presented approach yields a sufficiently accurate and patient-specific local SAR measurement of the brain within a scan time of less than 5 min.


Core (Polystyrene)-Shell [Poly(glycerol monomethacrylate)] Particles.
A set of water-swollen core-shell particles was synthesized by emulsion polymerization of a 1,3-dioxolane functional monomer in water. After removal of the 1,3-dioxolane group, the particles' shells were shown to swell in aqueous media. Upon hydrolysis, the particles increased in size from around 70 to 100-130 nm. A bicinchoninic acid assay and ζ-potential measurements were used to investigate the adsorption of lysozyme, albumin, or fibrinogen. Each of the core-shell particles adsorbed significantly less protein than the noncoated core (polystyrene) particles. Differences were observed as both the amount of difunctional, cross-linking monomer and the amount of shell monomer in the feed were changed. The core-shell particles were shown to be resistant to protein adsorption, and the degree to which the three proteins adsorbed was dependent on the formulation of the shell.


Second-site changes affect viability of amphotropic/ecotropic chimeric enveloped murine leukemia viruses.
Chimeras were previously generated between the ecotropic (Moloney-MuLV) and amphotropic (4070A) SU and TM proteins of murine leukemia virus (MuLV). After passage in D17 cells, three chimeras with junctions in the C terminus of SU (AE5, AE6, and AE7), showed improved kinetics of viral spreading, suggesting that they had adapted. Sequencing of the viruses derived from the D17 cell lines revealed second-site changes within the env gene. Changes were detected in the receptor binding domain, the proline-rich region, the C terminus of SU, and the ectodomain of TM. Second-site changes were subcloned into the parental DNA, singly and in combination, and tested for viability. All viruses had maintained their original cloned mutations and junctions. Reconstruction and passage of AE7 or AE6 virus with single point mutations recovered the additional second-site changes identified in the parental population. The AE5 isolate required changes in the VRA, the VRC, the VRB-hinge region, and the C terminus of SU for efficient infection. Passage of virus, including the parental 4070A, in D17 cells resulted in a predominant G100R mutation within the receptor binding domain. Viruses were subjected to titer determination in three cell types, NIH 3T3, canine D17, and 293T. AE6 viruses with changes in the proline-rich region initially adapted for growth on D17 cells could infect all cell types tested. AE6-based chimeras with additional mutations in the C terminus of SU could infect D17 and 293T cells. Infection of NIH 3T3 cells was dependent on the proline-rich mutation. AE7-based chimeras encoding L538Q and G100R were impaired in infecting NIH 3T3 and 293T cells.


Prevalence of Janus Kinase 2 mutations in patients with unusual site venous thrombosis.
We aimed to study patients with splanchnic vein thrombosis (SVT) and cerebral vein thrombosis (CVT) searching for JAK2 mutations. We evaluated 14 patients (median age: 41.5 years) with portal vein thrombosis (PVT) = 7; mesenteric vein thrombosis (MVT) = 3; and CVT = 4. JAK2 V617F was assessed by allele specific PCR of peripheral blood DNA. In addition, DNA was sequenced for other JAK2 mutations. Other inherited and acquired thrombophilia risk factors were evaluated. JAK2 V617F was positive in four out of seven patients with PVT and in one CVT patient. These five patients had a diagnosis of myelo-proliferative disorder (MPD) at the moment of the occurrence of thrombosis (n = 2) or later (n = 2). Patients with MVT and CVT were negative for JAK2 V617F, except one patient with CVT and a diagnosis of essential thrombocythemia. No other JAK2 mutations were found in this cohort. Besides MPD, other thrombophilia risk factors were identified in five patients. One patient had MPD as well as thrombophilia risk factor. In this group, 4 out of 7 of the patients with PVT carried the JAK2 V617F mutation with or without overt MPD. However, the investigation of other JAK2 mutations may not be necessary in patients with thrombosis at unusual sites.


Circular RNA and gene expression profiles in gastric cancer based on microarray chip technology.
The aim of the present study was to screen gastric cancer (GC) tissue and adjacent tissue for differences in mRNA and circular (circRNA) expression, to analyze the differences in circRNA and mRNA expression, and to investigate the circRNA expression in gastric carcinoma and its mechanism. circRNA and mRNA differential expression profiles generated using Agilent microarray technology were analyzed in the GC tissues and adjacent tissues. qRT-PCR was used to verify the differential expression of circRNAs and mRNAs according to the interactions between circRNAs and miRNAs as well as the possible existence of miRNA and mRNA interactions. We found that: i) the circRNA expression profile revealed 1,285 significant differences in circRNA expression, with circRNA expression downregulated in 594 samples and upregulated in 691 samples via interactions with miRNAs. The qRT-PCR validation experiments showed that hsa_circRNA_400071, hsa_circRNA_000543 and hsa_circRNA_001959 expression was consistent with the microarray analysis results. ii) 29,112 genes were found in the GC tissues and adjacent tissues, including 5,460 differentially expressed genes. Among them, 2,390 differentially expressed genes were upregulated and 3,070 genes were downregulated. Gene Ontology (GO) analysis of the differentially expressed genes revealed these genes involved in biological process classification, cellular component classification and molecular function classification. Pathway analysis of the differentially expressed genes identified 83 significantly enriched genes, including 28 upregulated genes and 55 downregulated genes. iii) 69 differentially expressed circRNAs were found that might adsorb specific miRNAs to regulate the expression of their target gene mRNAs. The conclusions are: i) differentially expressed circRNAs had corresponding miRNA binding sites. These circRNAs regulated the expression of target genes through interactions with miRNAs and might become new molecular biomarkers for GC in the future. ii) Differentially expressed genes may be involved in the occurrence of GC via a variety of mechanisms. iii) CD44, CXXC5, MYH9, MALAT1 and other genes may have important implications for the occurrence and development of GC through the regulation, interaction, and mutual influence of circRNA-miRNA-mRNA via different mechanisms.


A mannose receptor is involved in retinal phagocytosis.
A 175-kD mannose-specific receptor has been described in macrophages which appears to mediate pinocytosis and phagocytosis. A mannose-specific receptor has also been found on retinal pigment epithelial cells (RPE). Its role was examined in the phagocytosis of photoreceptor outer segments (ROS) by the RPE. All testing was done on cultured RPE cells challenged with fluorescein isothiocyanate (FITC)-labeled ROS for 4 hr at 37 degrees C. Total (internalized and bound) and phagocytized (internalized) ROS were quantified, and the effects of several experimental conditions on ROS phagocytosis were examined. Incubation of RPE cells in the presence of rabbit anti-serum (1:100) raised against the rat alveolar macrophage mannose receptor (anti-Mr) showed a 80% reduction in ROS phagocytosis compared with RPE in the presence of ROS alone. Anti-Mr preabsorbed with purified human mannose receptor protein (4 micrograms) showed no reduction in ROS phagocytosis. Incubation of RPE with preimmune serum showed no reduction in ROS phagocytosis. When FITC-labeled ROS (1.8 X 10(7)) were preincubated with purified mannose receptor, there was a 93% reduction in phagocytosis. Immunoblots of solubilized rat RPE microvillus membranes stained with the anti-Mr showed a single stained band at the 175-kD region, and immunolocalization studies showed specific labeling along RPE microvilli. These results suggest that a mannose-specific receptor is involved in retinal phagocytosis.


Prognostic significance of the tumour-adjacent tissue in head and neck cancers.
Even with significant advances in operative skills and adjuvant therapies, the overall survival of patients suffering with head and neck squamous cancers (HNSCC) is unsatisfactory. Accordingly, no clinically useful prognostic biomarkers have been found yet for HNSCC. Many studies analysed the expression of potential markers in tumour tissues compared to adjacent tissues. Nevertheless, due to the sharing of the same microenvironment, adjacent tissues show molecular similarity to tumour tissues. Thus, gene expression patterns of 94 HNSCC tumorous tissues were compared with 31 adjacent tissues and with 10 tonsillectomy specimens of non-cancer individuals. The genes analysed at RNA level using quantitative RT-PCR and correlated with clinico-pathological conditions were as follows: EGF, EGFR, MKI67, BCL2, BAX, FOS, JUN, TP53, VEGF, FLT1, MMP2, MMP9, MT1A and MT2A. The elevated MT2A, BAX, EGF and JUN expression was associated with the influence of tumour cells on the rearrangement of healthy tissues, as well as a significant shift in the BAX/BCL2 ratio. Our investigation also indicated that adjacent tissues play an important role in cancerogenesis by releasing several tumour-supporting factors such as EGF. A gradual increase in the metallothionein expression, from the lowest one in tonsillectomy samples to the highest ones in tumour samples, suggests that MT expression might be tissue reaction to the presence of tumour cells. The results of this study confirmed the significance of metallothionein in tumori-genesis and gave evidences for its use as a potential HNSCC biomarker. Furthermore, this study highlighted the importance of histologically normal tumour-adjacent tissue in prediction of HNSCC progress.


Segregational fidelity of chromosomes in human thyroid tumour cells.
Using fluorescence in situ hybridisation (FISH) we have analysed the segregational fidelity of all the human chromosomes during mitotic cell division. The losses and gains of chromosomes were analysed in human polyploid cell lines derived from a well-differentiated papillary thyroid cancer. These thyroid cells can be cultured for more than 300 population doublings. For the purpose of our study the polyploid nature of the cells may act as a protective buffer against the cell-lethal effects of the loss of individual chromosomes. To evaluate the role of the p53 gene product in maintaining the fidelity of chromosome segregation we compared the frequencies of chromosome loss and gain in cultures with wild-type p53 activity (K1E7neo3) and cultures transfected with plasmids expressing a mutant p53 product (K1E7scx6). Cultures were analysed for the presence of both structurally normal and rearranged chromosomes at both early and late passages. Cell cultures with defective p53 activity showed progressive chromosome loss from a median chromosome number of 87-97 to 75-86. Cell growth in cultures with wild-type p53 activity showed the loss of chromosomes 6, 7, and 8 and the gain of 17 and 20. Cultures expressing mutant p53 activity showed the loss of chromosomes 2, 5, 14 and 17 and the gain of 4 and 22. The combination of defective p53 and growth resulted in further destabilisation with the additional losses of chromosomes 3, 11, 15, 16 and 21. Chromosomes 1, 9, 10, 12, 13, 18, 19, X and Y segregated stably under all the culture conditions as did the structurally rearranged marker chromosomes. The study has demonstrated variation in the fidelity of mitotic chromosome segregation and the influence of p53 gene activity upon the segregation of individual human chromosomes.


[Imaging diagnosis of alveolar echinococcosis].
Alveolar echinococcosis, a parasitic disease caused in humans by the larval stage of Echinococcus granulosus tapeworm is a rare disease in Poland There are about 16 officially registered cases and further 8 cases described before creation of the European Registration system. This disease still causes significant diagnostic difficulties. Presently available radiological imaging techniques which can be applied in the alveolar echinococcosis diagnosis are presented. Ultrasound, CT and MRI images of patient with recurrent liver alveococcosis are presented against the background of literature data.


Effect of nesiritide (B-type natriuretic peptide) and dobutamine on ventricular arrhythmias in the treatment of patients with acutely decompensated congestive heart failure: the PRECEDENT study.
Dobutamine is commonly used as a means of treating decompensated congestive heart failure (CHF). Although typically effective at improving short-term hemodynamics and symptomatology, the frequent occurrence of arrhythmias and tachycardia is undesirable. In this randomized, multicenter trial, we compared the safety and clinical effectiveness of the cardiac hormone nesiritide (human B-type natriuretic peptide) with dobutamine in hospitalized patients with decompensated CHF. The study population consisted of 255 patients who were randomized to 1 of 2 doses of intravenous nesiritide (0.015 or 0.03 microg/kg/min) or dobutamine (> or =5 microg/kg/min) and stratified by means of an earlier history of ventricular tachycardia. Patients were also assessed with 24 hour Holter recordings immediately before and during study drug therapy and by means of signs and symptoms of CHF. Dobutamine significantly increased the mean (1) number of ventricular tachycardia events per 24 hours by 48 +/- 205 (P =.001), (2) repetitive ventricular beats per hour by 15 +/- 53 (P =.001), (3) premature ventricular beats per hour by 69 +/- 214 (P =.006), and (4) heart rate by 5.1 +/- 7.7 beats per minute (P <.001). These end points were significantly decreased or unchanged in the nesiritide groups. Nesiritide did not increase heart rate, despite a greater reduction of blood pressure. Both drugs were similarly effective means of improving signs and symptoms of CHF. Dobutamine is associated with substantial proarrhythmic and chronotropic effects in patients with decompensated CHF, whereas nesiritide actually reduces ventricular ectopy or has a neutral effect. Compared with dobutamine, nesiritide may be a safer, short-term treatment for patients with decompensated CHF.


Feline ilial fractures: assessment of screw loosening and pelvic canal narrowing after lateral plating.
To report the incidence of (1) screw loosening and (2) severity of pelvic canal narrowing in cats after repair of ilial fractures by a lateral bone plate. Case series. Cats (n=21) with ilial fractures. Medical records (June 1994 to February 2004) and radiographs were reviewed. Retrieved data were: signalment, fracture description, screw purchase, screw loosening, complications, and clinical outcome. Sacral index (SI) was used to objectively assess degree of pelvic canal narrowing. Long-term follow-up was by owner telephone interview or postal questionnaire. Screw loosening occurred in 13 cats (62%), with a mean loss of total screw purchase of 13% (range, 0-46%). Pelvic canal narrowing (mean decrease, 25%; range, 5-60%) occurred in all cats. Six cats (28%) had severe pelvic canal narrowing (mean, 52%); 5 of these had signs of recurrent constipation within 12 months postoperatively. Mean loss of total screw purchase in cats with severe pelvic narrowing was 28% compared with 9% for all other cats with narrowing. Lateral plating of ilial fractures in cats is associated with a high incidence of screw loosening. Pelvic canal narrowing >45% is associated with a high risk of recurrent constipation.


Sucrose clusters exhibiting a magic number in dilute aqueous solutions.
We report clustering of sucrose molecules in dilute aqueous solutions, based on measurements of the electrical mobility spectrum of singly charged airborne clusters produced by electrospraying the solution. The spectrum contains peaks with a smooth envelope, except for one peak which has about twice the amplitude of the envelope. The mobility at this peak is found to be somewhat higher than would be expected based on the rest of the sequence, suggesting a more compact structure for this cluster. This "magic" peak is prominent for basic, but not acidic, solutions. It is argued that these observations demonstrate the existence of clusters of many sizes in the solution prior to electrospraying, including the "magic" cluster. A simplified model is presented which reproduces the observed features.


The intestinal nematode inhibits T-cell reactivity by targeting P-GP activity.
Host immunosuppression occurs during chronic nematode infection, partly due to effector T-cell hyporesponsiveness. The role of P-glycoprotein (P-gp), a member of the ABC transporter family, has been assessed in T-cell activity. This study assesses the possible role of P-gp in T-cell activity during nematode infection. Our findings indicate that blockade of P-gp in vivo increased protection against Heligmosomoides polygyrus nematode infection and was associated with the enhanced T-cell activity. Three P-gp-inhibitors, verapamil (VRP), cyclosporine (CsA) and tariquidar (XR9576), were used to determine the influence of nematode infection on the P-gp function of T cells. The influence of the nematode on the uptake, efflux and kinetics of extrusion in T-cell subsets CD4+ and CD8+ was assessed by the accumulation of Rho123 dye. The results indicate that H. polygyrus infection contributes to the inhibition of T-cell function by elevating P-gp activity. The blockade of P-gp in the T cells of infected mice led to an impressive increase in T-cell proliferation and IL-4 cytokine release through the upregulation of NF-κB activation. These results provide the first evidence that the P-gp function of T cells is altered during nematode infection to open the way for further studies aiming to explore the role of P-gp in host-parasite interactions.


The effect of growth in 99.8 deuterium oxide on the ultrastructure of winter rye.
A preliminary report dealing with the ultrastructural effects of culture in a 99.8% D(2)O (deuterium oxide) environment on winter rye (Secale cereale L. cv. Winter) is presented. In general, the cells of D(2)O-cultured seedlings appeared similar to the cells of H(2)O-cultured seedlings. However, differences were found in chloroplast and dictyosome morphology, and ribosome number.


Comparative genomics for non-O1/O139 Vibrio cholerae isolates recovered from the Yangtze River Estuary versus V. cholerae representative isolates from serogroup O1.
Vibriocholerae, which is autochthonous to estuaries worldwide, can cause human cholera that is still pandemic in developing countries. A number of V. cholerae isolates of clinical and environmental origin worldwide have been subjected to genome sequencing to address their phylogenesis and bacterial pathogenesis, however, little genome information is available for V. cholerae isolates derived from estuaries, particularly in China. In this study, we determined the complete genome sequence of V. cholerae CHN108B (non-O1/O139 serogroup) isolated from the Yangtze River Estuary, China and performed comparative genome analysis between CHN108B and other eight representative V. cholerae isolates. The 4,168,545-bp V. cholerae CHN108B genome (47.2% G+C) consists of two circular chromosomes with 3,691 predicted protein-encoding genes. It has 110 strain-specific genes, the highest number among the eight representative V. cholerae whole genomes from serogroup O1: there are seven clinical isolates linked to cholera pandemics (1937-2010) and one environmental isolate from Brazil. Various mobile genetic elements (such as insertion sequences, prophages, integrative and conjugative elements, and super-integrons) were identified in the nine V. cholerae genomes of clinical and environmental origin, indicating that the bacterium undergoes extensive genetic recombination via lateral gene transfer. Comparative genomics also revealed different virulence and antimicrobial resistance gene patterns among the V. cholerae isolates, suggesting some potential virulence factors and the rising development of resistance among pathogenic V. cholerae. Additionally, draft genome sequences of multiple V. cholerae isolates recovered from the Yangtze River Estuary were also determined, and comparative genomics revealed many genes involved in specific metabolism pathways, which are likely shaped by the unique estuary environment. These results provide additional evidence of V. cholerae genome plasticity and will facilitate better understanding of the genome evolution and pathogenesis of this severe water-borne pathogen worldwide.


Capturing embryonic development from metamorphosis: how did the terminal patterning signalling pathway of Drosophila evolve?
The Torso receptor tyrosine kinase has two crucial roles in Drosophila melanogaster development. One is in the control of insect moulting, which is regulated by the neuropeptide hormone PTTH (prothoracicotropic hormone). PTTH activates ERK signalling via Torso in the prothoracic gland to stimulate ecdysone secretion. Torso also has a role in control of one of the earliest events in embryogenesis in Drosophila; patterning of the embryonic termini. Here Torso is activated by a different, but related, peptide called Trunk. During terminal patterning another protein, Torso-like, has a key role in mediating activation of Torso by Trunk. Torso-like is also expressed in the prothoracic gland and null-mutants have defective developmental timing in Drosophila. This function, however, has been recently shown to be independent of Torso and PTTH. We refer to these proteins, Trunk, PTTH, Torso and Torso-like, as the Torso-activation module. Outside Drosophila we see that the genes encoding the Torso-activation module have a complex phylogenetic history, with different origins and multiple losses of components of this signalling pathway during arthropod evolution. This, together with expression and functional data in a range of insects, leads us to propose that the terminal patterning pathway in Drosophila and Tribolium arose through co-option of PTTH/Trunk and Torso, which has a role in developmental timing, into a new context, and that Torso-like was recruited specifically in the ovary to modulate the specificity of this pathway.


Investigation of the intracellular stability and formation of a triple helix formed with a short purine oligonucleotide targeted to the murine c-pim-1 proto-oncogene promotor.
In our previous work we have shown that the oligonucleotide 5'-GGGGAGGGGGAGG-3' gives a very stable and specific triplex with the promoter of the murine c-pim-1 proto-oncogene in vitro[Svinarchuk, F., Bertrand, J.-R. and Malvy, C.(1994)Nucleic Acids Res., 22, 3742-3747]. In the present work, we have tested triplex formation with some derivatives of this oligonucleotide which are designed to be degradation-resistant inside the cells, and we show that phosphorothioate and the oligonucleotide with a 3' terminal amino group are still able to form triplexes. Moreover these oligonucleotides, like the 13mer oligonucleotide of similar composition [Svinarchuk, F., Paoletti, J., and Malvy, C. (1995) J. Biol. Chem., 270, 14068-14071], are able to stabilize the targeted duplex. In vivo DMS footprint analysis after electroporation of the pre-formed triplex into the cell have shown the presence of the triple helix inside the cells. This triplex structure partially blocks c-pim-1 promotor activity as shown by transient assay with a c-pim-1 promoter-luciferase gene construct. To our knowledge these data are the first direct evidence that conditions inside cells are favorable for triplex stability with non-modified oligonucleotides. However we were unable to show triplex formation inside living cells using various methods of oligonucleotide delivery. We suppose that this may be due to the oligonucleotide being sequestered by cellular processes or proteins. Further work is needed to find oligonucleotide derivatives and ways of their delivery to overcome the problem of triplex formation inside the cells.


Mutations in the zinc-binding motif of the reovirus capsid protein delta 3 eliminate its ability to associate with capsid protein mu 1.
Reovirus capsid protein delta 3 binds both double-stranded RNA (dsRNA) and zinc. Previous studies have revealed that the amino-terminal zinc finger is not required for the ability of delta 3 to bind dsRNA. We expressed wild-type and mutant delta 3 molecules by in vitro transcription/translation to evaluate the importance of the zinc finger for other functions of delta 3. delta 3 molecules with mutations in the zinc finger did not form complexes with capsid protein mu 1 but bound dsRNA more efficiently than wild-type delta 3 did. In contrast, a dsRNA-binding mutant was unimpaired in its ability to associate with mu 1. Studies with delta 3 fragments support these findings and indicate that sequences critical for delta 3's interaction with mu 1 lie in the amino terminus of the molecule. Our finding that mu 1 and dsRNA do not compete for identical binding sites on delta 3 has implications for its function as a translational regulator in infected cells.


Assessment of Erythrobacter Species Diversity through Pan-Genome Analysis with Newly Isolated Erythrobacter sp. 3-20A1M.
Erythrobacter species are extensively studied marine bacteria that produce various carotenoids. Due to their photoheterotrophic ability, it has been suggested that they play a crucial role in marine ecosystems. It is essential to identify the genome sequence and the genes of the species to predict their role in the marine ecosystem. In this study, we report the complete genome sequence of the marine bacterium Erythrobacter sp. 3-20A1M. The genome size was 3.1 Mbp and its GC content was 64.8%. In total, 2998 genetic features were annotated, of which 2882 were annotated as functional coding genes. Using the genetic information of Erythrobacter sp. 3-20A1M, we performed pangenome analysis with other Erythrobacter species. This revealed highly conserved secondary metabolite biosynthesis-related COG functions across Erythrobacter species. Through subsequent secondary metabolite biosynthetic gene cluster prediction and KEGG analysis, the carotenoid biosynthetic pathway was proven conserved in all Erythrobacter species, except for the spheroidene and spirilloxanthin pathways, which are only found in photosynthetic Erythrobacter species. The presence of virulence genes, especially the plant-algae cell wall degrading genes, revealed that Erythrobacter sp. 3-20A1M is a potential marine plant-algae scavenger.


Quantitative profiling of regional protein expression in rat retina after partial optic nerve transection using fluorescence difference two‑dimensional gel electrophoresis.
To examine the difference between primary and secondary retinal ganglion cell (RGC) degeneration, the protein expression at four regions of retina including superior, temporal, inferior and nasal quadrant in a rat model of partial optic nerve transection (pONT) using 2‑D Fluorescence Difference Gel Electrophoresis (DIGE) were investigated. Unilateral pONT was performed on the temporal side of optic nerves of adult Wistar rats to separate primary and secondary RGC loss. Topographical quantification of RGCs labeled by Rbpms antibody and analysis of axonal injury by grading of optic nerve damage at 1 week (n=8) and 8 weeks (n=15) after pONT demonstrated early RGC loss at temporal region, which is considered as primary RGC degeneration and progressing RGC loss at nasal region, which is considered as secondary RGC degeneration. Early protein expression in each retinal quadrant (n=4) at 2 weeks after pONT was compared with the corresponding quadrant in the contralateral control eye by DIGE. For all comparisons, 24 differentially expressed proteins (>1.2‑fold; P<0.05; ≥3 non‑duplicated peptide matches) were identified by mass spectrometry (MS). Interestingly, in the nasal retina, serum albumin and members of crystallin family, including αA, αB, βA2, βA3, βB2 and gamma S indicating stress response were upregulated. By contrast, only αB and βA2 crystallin proteins were altered in temporal quadrant. In the superior and inferior quadrants, βB2 crystallin, keratin type I, S‑arrestin and lamin‑B1 were upregulated, while heat shock cognate 71 kDa protein and heterogeneous nuclear ribonucleoproteins A2/B1 were downregulated. In summary, the use of DIGE followed by MS is useful to detect early regional protein regulation in the retina after localized optic nerve injury.


[Cloning and eukaryotic expression of human TRAF3IP3 gene].
To construct an eukaryotic expression plasmid of human TNF receptor-associated factor 3 in teracting protein 3(TRAF3IP3) gene and identify its expression in HEK293 cells. Human TRAF3IP3 cDNA was amplified by RT-PCR from bone marrow mononuclear cells. After digested by restriction enzymes XhoI and SalI, the complete open reading frame of TRAF3IP3 gene was inserted into pIRES2-EGFP eukaryotic expression vector with a Flag tag at the N-terminus. The recombinant plasmid was identified by double restriction enzyme digestion and sequencing analysis. The constructed TRAF3IP3 eukaryotic expression plasmid was transfected into HEK293 cells by calcium phosphate precipitation method. The expression of green fluorescence protein was observed by fluorescence microscopy. Real-time PCR and Western blotting were performed to detect the expression of Flag-TRAF3IP3 fusion protein. Double restriction enzyme digestion and sequencing analysis revealed that TRAF3IP3 eukaryotic expression plasmid was constructed successfully. Green fluorescence was detected in transfected HEK293 cells. Real-time PCR showed TRAF3IP3 mRNA was expressed at a high level. The approximate 62 kD Flag-TRAF3IP3 fusion protein was found by Western blotting. Human TRAF3IP3 eukaryotic expression plasmid pIRES2-EGFP-TRAF3IP3 has been constructed successfully, which provides a foundation for the gene function research of TRAF3IP3.


Model Selection and Model Averaging for Mixed-Effects Models with Crossed Random Effects for Subjects and Items.
A good deal of experimental research is characterized by the presence of random effects on subjects and items. A standard modeling approach that includes such sources of variability is the mixed-effects models (MEMs) with crossed random effects. However, under-parameterizing or over-parameterizing the random structure of MEMs bias the estimations of the Standard Errors (SEs) of fixed effects. In this simulation study, we examined two different but complementary perspectives: model selection with likelihood-ratio tests, AIC, and BIC; and model averaging with Akaike weights. Results showed that true model selection was constant across the different strategies examined (including ML and REML estimators). However, sample size and variance of random slopes were found to explain true model selection and SE bias of fixed effects. No relevant differences in SE bias were found for model selection and model averaging. Sample size and variance of random slopes interacted with the estimator to explain SE bias. Only the within-subjects effect showed significant underestimation of SEs with smaller number of items and larger item random slopes. SE bias was higher for ML than REML, but the variability of SE bias was the opposite. Such variability can be translated into high rates of unacceptable bias in many replications.


Real-world characteristics of hospitalized frail elderly patients with atrial fibrillation: can we improve the current prescription of anticoagulants?
In elderly patients, especially those older than 80 years, atrial fibrillation (AF) is associated with an almost 25% increased risk of stroke. Stroke prophylaxis with anticoagulants is therefore highly recommended. The prevalence of factors that have been associated with a lower rate of prescription and adherence to anticoagulant therapy in these patients is little known. The objective of this study was to explore the clinical characteristics of elderly subjects, with and without AF, consecutively admitted to an acute geriatric unit, discussing factors that may decrease the persistence on stroke prophylaxis therapy. We also highlight possible strategies to overcome the barriers conditioning the current underuse of oral anticoagulants in this segment of the population. A retrospective observational study was performed on a cohort of elderly patients with and without AF admitted to the Acute Geriatric Unit of San Gerardo Hospital (Monza, Italy). Compared to patients without AF (n = 1216), those with AF (n = 403) had a higher Charlson Comorbidity Index (3 vs. 2, P < 0.001), number of administered drugs (4 vs. 3, P < 0.001), rate of heart failure (36.5% vs. 12%, P < 0.001) and chronic kidney disease (20.6 vs. 13.2, P < 0.001). Many patients with AF were frail (54%) or pre-frail (29%). Elderly patients with AF have higher rates of conditions that affect adherence to traditional anticoagulant therapy (vitamin K antagonists, VKA). New direct oral anticoagulants (DOAs) can help overcome this problem. In order to prescribe the most appropriate VKA or DOAs, with the best efficacy/safety profile and the highest compliance, a comprehensive geriatric assessment should always accompany the scores for thrombotic and hemorrhagic risk stratification.


Isoenzyme patterns of soluble glutathione S-transferases from rat adrenal gland and ovary.
Soluble glutathione S-transferases (EC 188.8.131.52) of rat adrenal gland and ovary were separated by means of chromatofocusing. Four isoenzymes were found in the former and five in the latter organ. Most adrenal and ovarian GST are basic proteins with an apparent molecular mass of about 45 000. Some of them possess a weak delta 5-ketosteroid isomerase (EC 126.96.36.199) activity, too. The most abundant ovarian GST isoenzyme (iP = 9, 5) has common immunological determinants with the hepatic GST-B.


"We inform the experience of health": perspectives on professionalism in nursing self-employment.
Nursing work has evolved tremendously over the last century, raising ongoing questions about nursing's professional status. Through various strategies, professionalization in nursing has to some extent been accomplished, although autonomy over nursing practice has been elusive. This is especially so in the contemporary health care system, in which managerial control is emphasized and physician dominance continues. In response to professional constraints in traditional work settings, nursing self-employment is growing. In this study I used focused ethnography to explore the professional experiences of Canadian self-employed nurses and to reconsider nursing knowledge, ethics, and professionalism in this unique context. Despite the barriers they faced, these nurses offered a perspective on nursing professionalism that transcends classic professional traits, showing how the concept of professionalism can be invoked not as a way to "prove" status but as a way to describe a sense of commitment and the contribution to societal well-being.


EEG Correlates of Sustained Attention Variability during Discrete Multi-finger Force Control Tasks.
The neurophysiological characteristics of sustained attention states are unclear in discrete multi-finger force control tasks. In this article, we developed an immersive visuo-haptic task for conducting stimulus-response measurements. Visual cues were randomly provided to signify the required amplitude and tolerance of fingertip force. Participants were required to respond to the visual cues by pressing force transducers using their fingertips. Response time variation was taken as a behavioral measure of sustained attention states during the task. 50% low-variability trials were classified as the optimal state and the other high-variability trials were classified as the suboptimal state using z-scoring over time. A 64-channel electroencephalogram (EEG) acquisition system was used to collect brain activities during the tasks. The haptics-elicited potential amplitude at 20 ∼ 40 ms in latency and over the frontal-central region significantly decreased in the optimal state. Furthermore, the alpha-band power in the spectra of 8 ∼ 13 Hz was significantly suppressed in the frontal-central, right temporal, and parietal regions in the optimal state. Taken together, we have identified neuroelectrophysiological features that were associated with sustained attention during multi-finger force control tasks, which would be potentially used in the development of closed-loop attention detection and training systems exploiting haptic interaction.


Varidans streptococci in the oral flora of the patients at risk for infective endocarditis: species and penicillin susceptibilities.
This study examined viridans streptococci (SV) in oral flora of patients at risk of infective endocarditis (IE) by obtaining 57 isolates from gum-tooth margin swabs of 3 groups of patients. Penicillin minimal inhibitory concentrations (MICs) were determined by E test. Group 1 was from 30 patients with prosthetic heart valves who did not receive antimicrobial agents within 3 months prior to recruitment. Group 2 consisted of 21 patients with known rheumatic heart diseases who regularly received penicillin prophylaxis. Group 3 was 2 patients with IE caused by SV in whom dental swabs were performed before and on the third day of treatment. Streptococcus mitis was found most frequently (49% among 51 oral isolates of SV from patients in group 1 and 2) and in both blood cultures of IE patients. Ninety-four per cent of the isolates were penicillin-susceptible and the rest were intermediate-resistant (IR) equally distributed in both groups 1 and 2. In 2 patients with IE, oral SV obtained after 3 days of penicillin therapy had MICs rising 3 and 5 folds of the baselines. It is suggested that surveillance of susceptibilities of oral SV in patients at risk for IE should be kept up since this will affect the dose and type of antimicrobial agents in IE prophylaxis.


A synergistic effect of extracellular hypocalcemic condition for hyperoxic reoxygenation injury in rat hepatocytes.
Calcium accumulation of cells and mitochondria during reperfusion or reoxygenation has been implicated as a potential factor in cell injury as the result of mitochondrial damage. The objective of this study was to disclose whether or not low extracellular calcium ion concentration ([Ca2+]ex) in the medium at the time of reoxygenation might prevent calcium accumulation and attenuate hepatocytes injury after severe hypoxia. Isolated rat hepatocytes were incubated under a hyperoxic or hypoxic atmosphere for 60 min. During the ensuing 60-min hyperoxic reoxygenation, medium [Ca2+]ex was varied from 0.6 microM to 2.0 mM by altering total calcium and addition of chelators. Incubation in low [Ca2+]ex reduced total cellular calcium and mitochondrial calcium in both the hyperoxic and hypoxic group. Under hyperoxic/hyperoxic incubation (control), hepatocytes were able to maintain potassium balance when [Ca2+]ex was >3.0 microM (pCa=5.5) and cellular viability (% lactate dehydrogenase release) at all levels of extracellular calcium. Under hypoxic/hyperoxic incubation (reoxygenation), however, loss of the ability to restore potassium balance as well as apparent increase in lactate dehydrogenase release were observed at severely low [Ca2+]ex (<30 microM; pCa=4.5). This low [Ca2+]ex-induced exacerbation of hepatocytes viability could not be generated under mild reoxygenation such as normoxia. In normal isolated hepatocytes, very low [Ca2+]ex levels produce only very subtle changes in membrane permeability of isolated hepatocytes. After hypoxia, however, hypocalcemia acts synergistically with hyperoxic reoxygenation to produce more severe damage. These results suggested that [Ca2+]ex should be maintained on the physiological level to attenuate hepatocytes injury after severe hypoxia.


Endocrine and respiratory responses to ergotamine in Brahman and Hereford steers.
Ergot alkaloids are considered causative agents of fescue toxicosis, a syndrome experienced by cattle consuming tall fescue (Festuca arundinacea) infected with the fungal endophyte Neotyphodium coenophialum. One sign of fescue toxicosis in cattle is severe hyperthermia. This study assessed hormonal responses to ergotamine in heat-sensitive and heat-tolerant cattle. Seven Hereford (heat-sensitive, Bos taurus) and 7 Brahman (heat-tolerant, Bos indicus) steers on a fescue-free diet received ergotamine tartrate iv. Blood was sampled every 15 min for 2 h before and 4 h after dosing for determination of circulating hormonal changes. Respiration rates were recorded hourly. Ambient temperature and relative humidity averaged 31C and 48%, respectively, during sampling. A breed x time interaction existed (p < 0.01) for plasma prolactin, LH, insulin, glucagon, cortisol, triiodothyronine and glucose concentrations. The breed x time interaction tended to affect (p = 0.14) growth hormone and influenced (p < 0.01) respiration rates. Ergotamine reduced (p < 0.01) plasma LH and increased (p < 0.01) growth hormone concentrations in Brahman. Both breeds responded to ergotamine with increased (p < 0.01) plasma cortisol, glucagon, and glucose and reduced (p < 0.01) insulin concentrations. The magnitude of response for cortisol, insulin, and glucose were greater for the Brahman. Ergotamine increased (p < 0.01) plasma triiodothyronine and respiration rates in Hereford, whereas these traits were unaltered in Brahman. Acute ergotamine exposure generally resulted in similar effects on Brahman and Hereford steers. The triiodothyronine and respiratory comparisons revealed modified responses in Brahman that suggest a potential benefit of using heat-tolerant genetics to reduce the adverse effects of fescue toxicosis in cattle.


Phenotypic characterization of OmpX, an Ail homologue of Yersinia pestis KIM.
The goal of this study was to characterize the Yersinia pestis KIM OmpX protein. Yersinia spp. provide a model for studying several virulence processes including attachment to, and internalization by, host cells. For Yersinia enterocolitica and Yersinia pseudotuberculosis, Ail, YadA and Inv, have been implicated in these processes. In Y. pestis, YadA and Inv are inactivated. Genomic analysis of two Y. pestis strains revealed four loci with sequence homology to Ail. One of these genes, designated y1324 in the Y. pestis KIM database, encodes a protein designated OmpX. The mature protein has a predicted molecular mass of 17.47 kDa, shares approximately 70 % sequence identity with Y. enterocolitica Ail, and has an identical homologue, designated Ail, in the Y. pestis CO92 database. The present study compared the Y. pestis KIM6(+) parental strain with a mutant derivative having an engineered disruption of the OmpX structural gene. The parental strain (and a merodiploid control strain) expressed OmpX at 28 and 37 degrees C, and the protein was detectable throughout all phases of growth. OmpX was required for efficient adherence to, and internalization by, cultured HEp-2 cell monolayers and conferred resistance to the bactericidal effect of human serum. Deletion of ompX resulted in a significantly reduced autoaggregation phenotype and loss of pellicle formation in vitro. These results suggest that Y. pestis OmpX shares functional homology with Y. enterocolitica Ail in adherence, internalization into epithelial cells and serum resistance.


Tunable detection sensitivity of opiates in urine via a label-free porous silicon competitive inhibition immunosensor.
Currently, there is need for laboratory-based high-throughput and reliable point-of-care drug screening methodologies. We demonstrate here a chip-based label-free porous silicon (PSi) photonic sensor for detecting opiates in urine. This technique provides a cost-effective alternative to conventional labeled drug screening immunoassays with potential for translation to multiplexed analysis. Important effects of surface chemistry and competitive binding assay protocol on the sensitivity of opiate detection are revealed. Capability to tune sensitivity and detection range over approximately 3 orders of magnitude (18.0 nM to 10.8 muM) was achieved by varying the applied urine specimen volume (100-5 muL), which results in systematic shifts in the competitive binding response curve. A detection range (0.36-4.02 muM) of morphine in urine (15 muL) was designed to span the current positive cutoff value (1.05 muM morphine) in medical opiate urine screening. Desirable high cross-reactivity to oxycodone, in addition to other common opiates, morphine, morphine-3-glucuronide, 6-acetyl morphine, demonstrates an advantage over current commercial screening assays, while low interference with cocaine metabolite was maintained. This study uniquely displays PSi sensor technology as an inexpensive, rapid, and reliable drug screening technology. Furthermore, the versatile surface chemistry developed can be implemented on a range of solid-supported sensors to conduct competitive inhibition assays.


Body surface area as a determinant of pharmacokinetics and drug dosing.
Body surface area (BSA) was introduced into medical oncology in order to derive a safe starting dose for phase I studies of anticancer drugs from preclinical animal toxicology data. It is not clear however, as to why dosing by BSA was extended to the routine dosing of antineoplastic agents. Several formulas exist to estimate BSA, but the formula derived by DuBois and DuBois is the one used in adult medical oncology. This formula was derived based on data from only nine patients; subsequent attempts to validate the formula have found the DuBois formula to either over or underestimate the actual determined BSA. While cardiac output does correlate with BSA, the relationship between BSA and other physiologic measures relevant for drug metabolism and disposition, such as, renal and hepatic function, is weak or nonexistent. Further only epirubicin, etoposide, and carboplatin have been studied to determine if dosing by BSA would reduce interpatient variability, and none of these drugs were found to have significant relationships between their pharmacokinetics and BSA. Future clinical trials of new agents should not presume that dosing based on BSA reduces interpatient variability. Studies should examine the role, if any, BSA has in dosing new chemotherapeutic agents in initial phase I studies.


New examination of the performance of the MetaMax I metabolic analyser with the Douglas-bag technique.
To examine the performance of the MetaMax® I metabolic analyser of Cortex Biophysik, running a recent version of the software (MetaSoft v. 1.11.05), using the Douglas-bag method as reference. First, 182 measurements of the maximal O(2) uptake on school children were analysed both by a former version of the software (MetaMax Analysis) and by a recent version (MetaSoft). In further experiments, seven grown-up subjects of different fitness levels cycled for 5 min at constant powers between 50 and 350 W while the O(2) uptake was measured simultaneously by the MetaMax I and the Douglas-bag method during the last minute of altogether 39 exercises. The calculated maximal O(2) uptake of the 182 school children was on average 3% lower when the data were reanalysed by MetaSoft than when analysed by the former version of the software. There was in addition a 2% variation. In further experiments on grown-up subjects the O(2) uptake reported by the MetaMax I did not differ from that reported by the Douglas-bag system when data for all subjects were pooled, and the random error was 4%. However, the relationships differed slightly between the subjects (p = 0.005), and consequently the random error within each subject was 2-3%. The respiratory exchange ratio (R-value) reported by the MetaMax I differed systematically from that of the control method. However, a reliable R-value could be calculated from the instrument's raw data. Our data suggest that the MetaMax I running MetaSoft reports unbiased values of the O(2) uptake and with moderate random error.


Calcium signaling at the endoplasmic reticulum: fine-tuning stress responses.
Endoplasmic reticulum (ER) calcium signaling is implicated in a myriad of coordinated cellular processes. The ER calcium content is tightly regulated as it allows a favorable environment for protein folding, in addition to operate as a major reservoir for fast and specific release of calcium. Altered ER homeostasis impacts protein folding, activating the unfolded protein response (UPR) as a rescue mechanism to restore proteostasis. ER calcium release impacts mitochondrial metabolism and also fine-tunes the threshold to undergo apoptosis under chronic stress. The global coordination between UPR signaling and energetic demands takes place at mitochondrial associated membranes (MAMs), specialized subdomains mediating interorganelle communication. Here we discuss current models explaining the functional relationship between ER homeostasis and various cellular responses to coordinate proteostasis and metabolic maintenance.


Inhalation anaesthesia at the extremes of age: geriatric anaesthesia.
No single anaesthetic technique is superior for all elderly patients, although the prognosis is improved if minor surgical procedures are performed with local anaesthesia rather than with general or major regional anaesthesia. If general anaesthesia is required, which specific inhalation anaesthetic can improve the prognosis of the geriatric patient? The low solubility of desflurane and its resistance to biodegradation would seem to recommend its use in the geriatric patient. However, these theoretical advantages remain to be adequately documented.


Characterization of an Allosteric Pocket in Zika Virus NS2B-NS3 Protease.
The NS2B-NS3 protease from Zika virus (ZIKV NS2B-NS3pro) cleaves the viral polyprotein, being essential for its replication and a therapeutic target. Inhibitors that target the active site of ZIKV NS2B-NS3pro have been developed, but they tend to have unfavorable pharmacokinetic properties due to their highly positive charge. Thus, the characterization of allosteric sites in this protease provides new strategies for inhibitor development. Here, we characterized a new allosteric pocket in ZIKV NS2B-NS3pro, analogous to the one previously described for the dengue virus protease. Molecular dynamics simulations indicate the presence of cavities around the residue Ala125, sampling protein conformations in which they are connected to the active site. This link between the residue Ala125 and the active site residues was reinforced by correlation network analysis. To experimentally verify the existence of this allosteric mechanism, we expressed and purified the Ala125Cys mutant of ZIKV NS2B-NS3pro and demonstrated that this variant is inhibited by the thiol-containing chemical probes 5,5'-dithiobis-(2-nitrobenzoic acid) and aldrithiol, which do not affect the activity of the wild-type protein. Inhibition of the mutant protein is reversed by the addition of strong reducing agents, supporting the involvement of Cys125 in covalent bond formation and enzyme inhibition. Together, our results provide experimental evidence for an allosteric pocket in ZIKV NS2B-NS3pro, in the region around Ala125, and computational insights on the structural connection between this region and the enzyme active site.


New antipsychotics: classification, efficacy, and adverse effects.
Compared to traditional neuroleptics, most of the new antipsychotics are characterized by a low extrapyramidal side effect (EPS) liability and varying antipsychotic efficacy. This topic is reviewed for four principal classes of new, established, and potential antipsychotics: (1) Antipsychotics such as sulpiride and remoxipride that block a subgroup of dopamine (DA) D2/D3 receptors produce a relatively low level of side effects, including EPS, and have an antipsychotic effect equal to or slightly weaker than traditional neuroleptics. D1 antagonists demonstrate a low level of EPS in primates and may prove to be a valuable new type of antipsychotic drug. (2) Theoretically, partial D2 agonists have the advantage of producing few or no EPS and a specific beneficial effect in negative symptoms, but as yet the expectations have not been fulfilled. (3) Nondopamine drugs such as serotonin (5HT1) agonists, 5HT2 antagonists, 5HT3 antagonists, and gamma-amino-butyric-acid-A (GABA-A) benzodiazepine agonists have anxiolytic, antidepressant, antiaggressive, and maybe antiparkinsonian effects and may play an adjunctive role in the treatment of schizophrenia. 5HT3 antagonists (e.g., ondansetron), partial benzodiazepine agonists, and partial glutamate agonists may prove to be effective antipsychotics. (4) Antipsychotics such as clozapine and risperidone, which affect D2/D3 receptors as well as 5HT, alpha 1, and/or D1 receptors appear to have the most pronounced antipsychotic effect.


Interleukins and redox impairment in type 2 diabetes mellitus: mini-review and pilot study.
Type 2 diabetes mellitus (T2DM) represents a leading cause of morbidity and premature mortality, low-grade inflammation being acknowledged as a key contributor to its development and progression. A tailored therapeutic approach, based on sensitive and specific biomarkers, could allow a more accurate analysis of disease susceptibility/prognostic and of the response to treatment. This mini-review and pilot study had two main goals: (1) reviewing the most recent literature encompassing the use of interleukins as inflammatory markers influenced by the redox imbalances in T2DM and (2) assessing parameters that conjunctly evaluate the redox impairment and inflammatory burden of T2DM patients, taking into consideration smoking status, as such group-specific biomarkers are scarcely reported in literature. Firstly, PubMed database was surveyed to select and review the relevant studies employing interleukins as T2DM biomarkers and to assess if studies using combined inflammatory-redox indices were reported. Then, routine biochemical parameters were assessed in a pilot study -T2DM patients with 3 subgroups: non-smokers, smokers and ex-smokers, were compared to a control group of non-diabetic, apparently healthy non-smokers. Protein (AOPPs, AGEs), lipid/HDL (Amplex Red-based method) oxidative damage and inflammatory status (CRP, IL-1β, IL-6, IL-10) biomarkers were assessed. Cytokine ratios and 2 oxidative-inflammatory status indices were developed (IH1 and IH2) and evaluated. We observed significant differences in terms of serum redox and inflammatory status (AOPPs, AGEs, CRP, CRP/HDL, CRP/IL-6, IL-10/IL-6, IH1) between T2DM patients compared to control and, moreover, between the subgroups formed considering smoking status (CRP, CRP/HDL, IH1). Glycemic control strongly influenced inflammatory status biomarkers: glycemia was positively correlated with the inflammatory parameters (CRP/IL-10) and inversely with the anti-inflammatory ones (IL-10, IL-10/IL-1β ratio). Several of the assessed parameters may possess prognostic value for diabetics, especially when comparing subgroups with a different smoking history and could prove useful in clinical practice for assessing disease progress and therapeutic efficacy.


[CLINICAL AND PSHYSIOLOGICAL FEATURES OF INTRAOSSEOUS ANESTHESIA DURING DENTAL IMPLANTATION].
The aim of research - to assess the feasibility and effectiveness, of the intraosseous method of anesthesia, with the Quick Sleeper injection system on the toothless area of the alveolar bone during dental implantation. Clinical and physiological studies were conducted to study the features of performing intraosseous anesthesia on the site of alveolar bone adentia in the absence of the usual anatomical topographic landmarks. New computer technologies, as well as modern methods of patient examination and observation were used. The results of the analysis confirmed the advantages of intraosseous anesthesia compared with alternative methods of local anesthesia during dental implantation. The advantages of the intraosseous method of anesthesia are characterized, the features of conducting, the anatomical guidelines have been indicated for an effective intraosseous anesthesia in the area of the alveolar bone adentia, during dental implantation.