Energy requirements for wet solvent extraction of lipids from microalgal biomass.
Biofuel production from microalgae requires energy efficient processes for extracting and converting triacylglyceride lipids to fuel, compatible with coproduction of protein feeds and nutraceuticals. Wet solvent extraction involves mechanical cell rupture, lipid extraction via solvent contacting, physical phase separation, thermal solvent recovery, and transesterification. A detailed analysis of the effect of key process parameters on the parasitic energy demand of this process was performed. On a well-to-pump basis, between 16% and 320% of the resultant biodiesel energy was consumed depending solely on the process parameters. Highly positive energy balances can be achieved, but only if a correctly designed process is used. This requires processing concentrated biomass (ca 25%w/w) with a high triacylglyceride content (ca 30%w/w), and an efficient extraction process employing a non-polar solvent, low solvent-to-paste ratio, and efficient energy recovery. These requirements preclude many laboratory scale processes and polar co-solvents as viable options for large-scale biofuel production.


Sound-evoked efflux of excitatory amino acids in the guinea-pig cochlea in vitro.
We have used the perfused guinea-pig temporal-bone preparation to study the sound-evoked efflux of aspartate and glutamate, which are putative afferent transmitters in the cochlea. The cochlea was stimulated with white noise at 89, 95, and 101 dB SPL. Cochlear function was monitored by recording the endocochlear potential, the cochlear microphonic, and the summating potential. In silence, there was a low basal efflux of both amino acids. A significant and intensity dependent sound-evoked efflux of aspartate was observed at all levels, whereas a significant efflux of glutamate was found only at the 101 dB SPL level. Immunohistochemistry of sections from the organ of corti showed an ubiquitous distribution of glutamate-like immunoreactivity in the sensory organ and ganglion, whereas aspartate-like immunoreactivity was found in the region of the inner hair cells and in the spiral ganglion. In view of these findings, we suggest that not only glutamate, but also aspartate may have a neurotransmitter role in the afferent pathway of the cochlea.


CT angiography: application to the evaluation of carotid artery stenosis.
To determine whether computed tomographic (CT) angiography can be used to accurately quantify carotid stenoses and differentiate severe stenoses from occlusions. CT angiography was used to evaluate 50 carotid arteries in 27 patients who were referred for surgical evaluation after diagnosis of carotid stenosis and who had undergone standard angiography. Four observers read each study separately in a blinded fashion, then all four observers together reviewed those studies in which the individual readings differed to reach a consensus reading. The consensus readings for the two modalities were in agreement on stenosis category in 41 (82%) of the 50 carotid arteries (kappa W = 0.852 +/- 0.114). The consensus readings in the other nine arteries differed by only one category; seven had less severe stenosis at CT angiography and two had more severe stenosis. The CT angiograms also depicted a variety of additional abnormalities, including loops (n = 6), aneurysms (n = 2), and ulcers (n = 4). These results indicate that CT angiography can non-invasively provide most of the information needed before carotid endarterectomy.


Estimating the opportunity costs of environmental conservation in the Feijão River watershed (São Carlos-SP, Brazil).
The objective of this study was to infer the opportunity cost of land use of the Feijão River watershed (São Carlos-SP, Brazil), in order to estimate the financial resources necessary to compensate landowners willing to convert their production areas into areas of environmental preservation. Net values were estimated by calculating the Annual Value and the Net Present Value of each activity. The area used for agricultural production was estimated using the Land Cover Map of the watershed. The study involved four production areas: forestry, livestock, sugarcane and orange, accounting for 66% of the watershed area of 22,300 hectares. Considering a scenario of total consent from landowners, the 2011 net annual values were estimated at R$ 13.4 million: R$ 2.2 million (eucalyptus), R$ 1.9 million (livestock), R$ 1.1 million (sugarcane) and R$ 8.2 million (orange). This amount would be used as payment for ecosystem services.


Lipopeptides as therapeutics: applications and in vivo quantitative analysis.
A number of novel lipopeptides have been studied for their possible therapeutic potential. These studies should be supported by the appropriate analytical tools not only for novel potential drugs but also for their metabolites, precursors and side products. Lipopeptides have specific physicochemical properties that make them successful in medical applications. However, there are some difficulties with their qualitative and quantitative analyses in biological samples. Therefore, reliable, sensitive and robust analytical methods are in high demand. The main interest of our review is to describe a selection of specific and important properties of lipopeptides, and the analytical methods currently utilized for their characterization and determination in biological samples. A comparison of the pros and cons of immunomethods versus LC-MS methods is discussed in detail.


Engraftment kinetics of human cord blood and murine fetal liver stem cells following in utero transplantation into immunodeficient mice.
This study was undertaken to evaluate the kinetics of engraftment after in utero transplantation of murine fetal liver and human cord blood stem cells in the nonobese diabetic/severe combined immunodeficient (NOD/SCID) mouse model. NOD/SCID fetuses were injected with murine fetal liver or human cord blood CD34+ cells at day 13.5 of gestation. Frequencies of donor cells were analyzed by flow cytometry up to 48 h post transplantation and 4-16 weeks postnatally. Hematopoietic multilineage reconstitution capacity was assessed. Both types of donor cells home rapidly. However, the frequency of human cord blood stem cells rapidly diminished while the murine fetal liver stem cells expanded over time, resulting in multilineage hematopoietic reconstitution. Differences in long-term reconstitution of allogeneic versus xenogeneic donor cells were ascribed to the inability of the human cells to self-renew and differentiate in the fetal mouse environment, demonstrating the limitations of this commonly used xenograph.


Workplace bullying and sickness presenteeism: cross-sectional and prospective associations in a 2-year follow-up study.
The aim of this study was to investigate exposure to workplace bullying as a potential risk factor for sickness presenteeism (SP), i.e. working while ill. This study is based on data collected through self-reported questionnaires in a 2-year prospective study on employees in Denmark. At baseline, 3363 employees (45.7 % response rate) answered to a questionnaire on their psychosocial work environment and health status. After 2 years, 1664 of the respondents also completed a follow-up questionnaire (49.5 % of the total baseline respondents). After excluding participants with missing values, the final samples were composed of 2865 and 1331 participants in the cross-sectional and prospective analyses, respectively. Modified poisson regression analyses showed that frequent (i.e. daily or weekly) exposure to workplace bullying was associated with reporting 8 or more days of SP in the preceding year in both the cross-sectional and the prospective analysis, also when controlling for several confounders including health-related variables. However, the prospective relationship became non-significant after adjustment for baseline SP. This study provides indications of a significant relationship between exposure to frequent workplace bullying and SP, although causal connections could not be established. Methodological and theoretical considerations about study findings are provided, which could be of benefit to future studies examining the impact of being a target of workplace bullying on SP.


Polymer-supported pyridine-bis(oxazoline). Application to ytterbium-catalyzed silylcyanation of benzaldehyde.
[reaction: see text] Terminal acetylenes containing hydroxy and carboxylic acid groups were subjected to Sonogashira coupling with 4-bromo-2,6-bis[(R)-4-phenyloxazolin-2-yl]pyridine and the resulting pybox derivatives were immobilized on Tentagel resins. Ytterbium(III) chloride complexes of the polymeric ligands catalyzed the addition of trimethylsilyl cyanide to benzaldehyde with 80-81% ee. The ligands were reused more than 30 times without any loss in selectivity or activity, and the metal complexes could be recovered and reused at least four times, although with slightly decreasing activity.


Area-level relative deprivation and alcohol use in Denmark: Is there a relationship?
Greater area-level relative deprivation has been related to poorer health behaviours, but studies specifically on alcohol use and abuse have been equivocal. The main purpose of the present study was to investigate how area-level relative deprivation in Denmark relates to alcohol use and misuse in the country. As individual-level data, we used the national alcohol and drug survey of 2011 ( n= 5133). Data were procured from Statistics Denmark to construct an index of relative deprivation at the parish level ( n=2119). The deprivation index has two components, which were divided into quintiles. Multilevel linear and logistic regressions analysed the influence of area deprivation on mean alcohol use and hazardous drinking, as measured by the Alcohol Use Disorder Identification Test. Men who lived in parishes designated as 'very deprived' on the socioeconomic component were more likely to consume less alcohol; women who lived in parishes designated as 'deprived' on the housing component were less likely to drink hazardously. But at the individual level, education was positively related to mean alcohol consumption, and higher individual income was positively related to mean consumption for women. Higher-educated men were more likely to drink hazardously. Area-level measures of relative deprivation were not strongly related to alcohol use, yet in the same models individual-level socioeconomic variables had a more noticeable influence. This suggests that in a stronger welfare state, the impact of area-level relative deprivation may not be as great. Further work is needed to develop more sensitive measures of relative deprivation.


Structural and immunological characterization of the intracellular forms of an abundant 68,000 Mr human cytomegalovirus protein.
Murine monoclonal antibodies and polyvalent monospecific antisera reactive with an abundant 68,000 Mr (p68) human cytomegalovirus (CMV) virion protein were used to characterize this protein within CMV-infected cells. The protein was found to partition within the nucleus and cytoplasm of infected cells. Pulse-chase analysis indicated the p68 was degraded into three proteins of 52,000, 51,000 and 50,000 Mr which were found only within infected cells. Both cellular forms as well as the virion p68 were phosphorylated but non-glycosylated. The p68 was synthesized shortly after infection and in the presence of cytosine arabinoside, an inhibitor of viral DNA replication. Studies with monospecific antisera and a panel of monoclonal antibodies specific for the p68 suggested that this protein was not expressed on the surface of infectious virions or infected cells.


Physician role conflict and resulting career changes. Gender and generational differences.
To evaluate gender and generational differences both in the prevalence of role conflict and in resulting career changes among married physicians with children. Cross-sectional survey. We sent a survey to equal numbers of licensed male and female physicians (1,412 total) in a Southern California county; of the 964 delivered questionnaires, 656 (68%) were returned completed. Our sample includes 415 currently married physicians with children, 64% male and 36% female. The prevalence of perceived role conflict, or career changes for marriage, and of career changes for children were evaluated. Types of career changes were also evaluated. More female than male physicians (87% vs 62%, p < .001) and more younger than older female physicians (93% vs 80%, p < .01) and male physicians (79% vs 54%, p < .001) experienced at least moderate levels of role conflict. Younger female and male physicians did not differ in their rates of career change for marriage (57% vs 49%), but female physicians from both age cohorts were more likely than their male peers to have made career changes for their children (85% vs 35%, p < .001). Younger male physicians were twice as likely as their older peers to have made a career change for marriage (49% vs 28%, p < .001) or children (51% vs 25%, p < .001). The most common type of career change made for marriage or children was a decrease in work hours. Most physicians experience role conflict, and many adjust their careers in response. Flexible career options may enable physicians to combine professional and family roles more effectively.


Proton and phosphorus-31 nuclear magnetic resonance spectroscopy of human bile in hepatopancreaticobiliary cancer.
Hepatopancreaticobiliary cancers can be difficult to diagnose. Nuclear magnetic resonance (NMR) spectroscopy provides non-invasive information on phospholipid metabolism, and previous studies of liver tissue have highlighted changes in phospholipids in malignancy. We hypothesised that in-vitro NMR spectroscopy of human bile may provide independent diagnostic indices in cancer management through an assessment of the phospholipid content. Bile samples from 24 patients were collected at endoscopic retrograde cholangiopancreatography and from one subject at cholecystectomy. Thirteen patients had cancer: pancreatic carcinoma (eight), cholangiocarcinoma (three) and metastatic liver disease (two). The remaining 12 patients had non-malignant pathology. In-vitro proton (H) and phosphorus-31 (P) NMR spectra were obtained from all samples using an 11.7 Tesla NMR spectroscopy system. Complementary information was obtained from the H and P NMR spectra. Signals were assigned to phosphatidylcholine in both H and P NMR spectra. Phosphatidylcholine levels were significantly reduced in the bile from cancer patients when compared with bile from non-cancer patients (P=0.007). These preliminary studies suggest that H and P NMR spectroscopy of bile may be used to detect differences in phospholipid content between cancer and non-cancer patients. This may have implications for the development of novel diagnostic strategies in hepatopancreaticobiliary cancers. Further larger-scale studies are warranted.


Algorithmic improvements to species delimitation and phylogeny estimation under the multispecies coalescent.
The focus of this article is a Bayesian method for inferring both species delimitations and species trees under the multispecies coalescent model using molecular sequences from multiple loci. The species delimitation requires no a priori assignment of individuals to species, and no guide tree. The method is implemented in a package called STACEY for BEAST2, and is a extension of the author's DISSECT package. Here we demonstrate considerable efficiency improvements by using three new operators for sampling from the posterior using the Markov chain Monte Carlo algorithm, and by using a model for the population size parameters along the branches of the species tree which allows these parameters to be integrated out. The correctness of the moves is demonstrated by tests of the implementation. The practice of using a pipeline approach to species delimitation under the multispecies coalescent, has been shown to have major problems on simulated data (Olave et al. in Syst Biol 63:263-271. doi: 10.1093/sysbio/syt106 , 2014). The same simulated data set is used to demonstrate the accuracy and improved convergence of the present method. We also compare performance with *BEAST for a fixed delimitation analysis on a large data set, and again show improved convergence.


[Maxillary sinus cancers: Prognosis and quality of life: a retrospective study of 22 cases].
Maxillary sinus cancer is uncommon. We present a study of 22 cases of patients managed at the Nantes University Hospital for maxillary sinus cancer during the time period 1989-2002. Clinical and oncological characteristics of the tumors were evaluated. Treatment and results were evaluated in association with the patients' quality of life, estimated with a questionnaire survey sent to surviving patients. Carcinomas predominated (68%). The TNM classification was: T2: 45.5%, T3: 45.5% and T4: 9%. 89% of necks were N0 and 9% were N1. Treatment course (determined after imaging) was surgical removal of the tumor by total maxillectomy: 77.5%; partial maxillectomy: 22.5%, associated with post-operative radiotherapy. Esthetic consequences and disabilities of patients assessed by the questionnaire pointed out that 68% of them were complaining of poor quality of life. Global survival at 12 months was 63%, 20% at 24 months and 9% at 5 years. There were no statistically significant differences between specific survivals according to T-stage nor according to histological features. The difference was significant when surgical removal of the tumour was large according to post-operative histology. Authors conclude that maxillary sinus cancers exhibit poor prognosis, and focus on the necessity of quality of life assessment.


Is the National Registration website (Australian Health Practitioner Regulation Agency) effective in tracking Australian medical graduates' rural work?
This study compared rural location identified through the National Registration (AHPRA) registry with location obtained through labour-intensive personal contact. Longitudinal cohort study with two methods to identify the work locations of medical graduates from The Rural Clinical School of Western Australia (RCSWA). Consenting alumni from the University of Western Australia and the University of Notre Dame Fremantle participating in RCSWA between 2002 and 2009 inclusive and available to contact in 2011. Percentage location matches between two contact methods. There was 80% agreement for principal suburb, 92% agreement for principal city and 94% agreement for principal state between RCSWA personal contact and the AHPRA registry. AHPRA identified nearly two times as many graduate locations. However, there was only 31% agreement for a rural placement location (of any length). In more detail, for year-long rural placement, personal contact was 88% concordant with AHPRA; work six months or more were less concordant (44% agreement); work less than six months were not concordant (4% agreement). AHPRA data matched RCSWA alumni data only for graduates in full-time rural work. Since medical alumni spend up to 10 years in pre-vocational and vocational training, which includes many rural options, personal contact was able to pick up the myriad of rural choices, whereas the AHPRA database was not sensitive enough to identify them. Until graduates have stably finished training, the optimal method to identify rural work is through personal contact but statistical correction for missing data needs to be considered.


Optimization of triarylimidazoles for Tie2: influence of conformation on potency.
In an effort to understand the effect of N-alkylation of triarylimidazoles on Tie2 inhibition, ortho-substituted C-2 aryl analogs were synthesized to investigate the effect of different torsion angles on potency. This exercise resulted in the identification of a potent and selective tetrasubstituted imidazole that was efficacious in an animal model of angiogenesis.


Process of technical stabilization of gastric endoscopic submucosal dissection at the National Cancer Center in Japan.
Endoscopic submucosal dissection (ESD) was originally developed in 1995 using an insulation-tipped diathermic knife (Olympus, Tokyo, Japan) to achieve en-bloc resection of early gastric cancer (EGC). It has been suggested that advances in endoscopic devices and medical equipment, such as the high-frequency generator (1999) and hemostatic forceps (2000), and procedural improvements including post-ESD preventive coagulation of visible vessels in the resection area (2003) led to further progress of ESD; therefore, we investigated the actual process of technical stabilization of gastric ESD. A total of 1,713 consecutive patients with solitary differentiated-type EGC at initial onset underwent ESD at our hospital from 1995 to 2006. We retrospectively assessed ESD outcomes for all patients by dividing them into three chronological periods: 1995-1998 (1st period; 57 patients), 1999-2002 (2nd period; 563 patients), and 2003-2006 (3rd period; 1,093 patients). The en-bloc resection, intraoperative bleeding, and delayed bleeding in the 1st/2nd/3rd period were 52.6%/94.7%/99.3% (1st vs. 2nd, p<0.01; 2nd vs. 3rd, p<0.01), 8.8%/7.1%/1.6% (1st vs. 2nd, no significant difference; 2nd vs. 3rd, p<0.01), and 15.8%/7.6%/3.3% (1st vs. 2nd, p<0.05; 2nd vs. 3rd, p<0.01), respectively. Gastric ESD has stabilized technically following advances in endoscopic devices and medical equipment together with procedural improvements.


Bartholinitis after vulvovaginal surgery.
The object of the study was to determine whether obstruction of the Bartholin gland duct after vulvovaginal surgery is a cause of Bartholin gland infections or cysts. The records of patients with vulvovaginal surgical procedures performed by me between 1983 and 1997 were reviewed for evidence of a Bartholin gland infection or cyst formation. A postoperative Bartholin gland infection occurred in 2 of 217 patients with posterior colporrhaphy (with or without vault suspension) and 1 of 14 patients with partial vestibulectomy for vestibulitis. No patient had a chronic Bartholin cyst develop. Bartholin gland infections and Bartholin cysts are uncommon occurrences after surgical procedures that have the potential to obstruct the Bartholin duct.


Children of alcoholics. Report of a preliminary study and comments on the literature.
Familial relationships in 11 families in which 1 parent is an alcoholic are discussed in the context of literature on the children of alcoholics.


Perturbation-free measurement of the harmonics-to-noise ratio in voice signals using pitch synchronous harmonic analysis.
The measurement of the harmonics-to-noise ratio (HNR) in speech signals gives an indication of the aperiodicity of the speech waveform. This may be due to the presence of jitter, shimmer, additive noise, waveshape change, or some unknown combination of these factors. In order to estimate the HNR as a measure of the additive noise component only, the contaminating effects of the other contributory components must first be removed. A pitch synchronous harmonic analysis is proposed to overcome this problem. The procedure takes advantage of the time scale compression-frequency expansion property of the Fourier series in order to eliminate jitter and shimmer. Successive spectra are added by harmonic number as opposed to frequency location, and perturbation is removed due to the fact that the relative heights of the harmonic components remain the same for scaled signals. The technique is examined on synthetically generated voice signals. A discussion of the results is given in terms of human voice signals, characterization of jitter, vocal tract filtering effects, perturbation mechanisms, nonlinear dynamics, and the possibility of developing the method for use with inverse filtering strategies.


[Histophysiology of the thymus gland].
The article comprises a survey of the literature data on histophysiology of the thymus gland and a discussion of results of the authors' own observations of this gland in rats line "Vistar" in normalcy and in the presence in the organism of a transplanted tumour RS-1. Problems associated with histophysiology of the thymus, including such as differentiation of thymocytes, their relationships with the reticuloepithelial cells, the capacity of the latter for secretion, the role of the stratified epithelial bodies, the hematothymus barrier, etc., are discussed.


Northward invasion of the parasitic deer ked ( Lipoptena cervi), is there geographical variation in pupal size and development duration?
The deer ked (Lipoptena cervi) is a common ectoparasite of cervids. During the last decades the species has rapidly invaded in northern Europe, especially in Finland, towards the north and increased its prevalence on the moose population. Consequently, during this rapid invasion the deer ked has faced more severe climatic conditions. We studied whether pupal size (measured as pupal weight) and pupal development duration of the deer ked varies along historical invasion zones and temperature zones towards north in Finland. Moreover, we explored possible size- and gender-dependent variation in pupal development duration. We divided wild-collected pupae in respect to their origin in two ways: (1) temperature zones (from south-west to colder north-east) and (2) invasion history (from early to late establishment). We reared pupae in the controlled laboratory conditions in identical temperature and light conditions. Pupal size decreased towards north and the smaller pupae developed faster. However, the results do not show differences in pupal size or developmental characteristics between the invasion zones. This supports the idea of rapid developmental plasticity of the deer ked and that not the invasion history but the current temperature regime determines the life history of the deer ked when invading towards a colder environment.


Risk factors for late-onset seizures related to cerebral contusions in adults with a moderate traumatic brain injury.
This prospective study compares the characteristics of patients with a moderately severe traumatic brain injury (TBI) and cerebral contusions who develop late-onset seizures to those who do not. Thirty-nine adult TBI patients with cerebral contusions, who did not need a neurosurgical treatment, could be followed up for more than 3years. Fourteen patients developed seizures during that period and 25 did not. The Glasgow Coma Scale (GCS) score on admission and the modified Rankin (mR) score on discharge from the hospital, the computed tomography (CT) and/or magnetic resonance imaging (MRI) findings, the electroencephalogram (EEG) patterns as well as the vascular and habit risk factors were compared between both groups. The mean GCS and mR scores were moderately severe and comparable between both groups. Early-onset seizures represented 21.4%. The overall seizure recurrence was 85.7% after treatment with carbamazepine or valproate sodium. Still 3 patients did not remain seizure-free after addition of another antiepileptic drug. The average number of brain contusions on CT/MRI was approximately the same. Vascular risk factors and alcohol abuse were more observed in the seizure patients. Abnormal EEG findings on discharge from the hospital were significantly more frequent in the patients who developed late-onset seizures afterward (P<0.05). Seizures after non-complicated cerebral contusions are difficult to treat. Vascular risk factors and alcohol abuse may also predispose to their occurrence. The EEG findings after the TBI are highly predictive.


Are optic disc haemorrhages a common finding in all glaucoma patients?
Disc haemorrhages are an important risk factor in development and progression of glaucomatous damage, but it is not known whether such bleedings can be found in all glaucoma patients. In this retrospective study of 1506 patients with manifest or suspected glaucoma, sequential optic disc stereophotographs revealed a disc haemorrhage in 94 cases. Distribution of the haemorrhagic and non-haemorrhagic patients according to the frequency of photography indicates that the 2 groups of patients represent different glaucoma populations. Furthermore, patients who do not show a haemorrhage in 1 or 2 random examinations probably will not show a bleeding in the later follow-up, either.


Clinical pharmacokinetics of almotriptan, a serotonin 5-HT(1B/1D) receptor agonist for the treatment of migraine.
The pharmacokinetics of almotriptan are linear over a range of oral doses up to 200mg in healthy volunteers. The compound has a half-life of approximately 3 hours. Almotriptan is well absorbed after oral administration and the mean absolute bioavailability is 69.1%. Maximal plasma concentrations are achieved between 1.5 and 4 hours after dose administration; however, within 1 hour after administration, plasma concentrations are approximately 68% of the value at 3 hours after administration. Food does not significantly affect almotriptan absorption. Almotriptan is not highly protein bound and is extensively distributed in the body. Approximately 50% of an almotriptan dose is excreted unchanged in the urine; this is the predominant single mechanism of elimination. Renal clearance is mediated, in part, through active tubular secretion, while the balance of the almotriptan dose is metabolised to inactive compounds. The predominant route of metabolism is via monoamine oxidase-A, and cytochrome P450 (CYP) mediated oxidation (via CYP3A4 and CYP2D6) occurs to a minor extent. Almotriptan clearance is moderately reduced in elderly subjects, but the magnitude of this effect does not warrant a dose reduction. Sex has no significant effect on almotriptan pharmacokinetics. Almotriptan pharmacokinetic parameters do not differ between adolescents and adults, and absorption is not affected during a migraine attack. As expected, renal dysfunction results in reduced clearance of almotriptan. Patients with moderate-to-severe renal dysfunction should use the lowest dose of almotriptan and the total daily dose should not exceed 12.5 mg. Similar dosage recommendations are valid for patients with hepatic impairment, based on the clearance mechanisms for almotriptan. Drug-drug interaction studies were conducted between almotriptan and the following compounds: fluoxetine, moclobemide, propranolol, verapamil and ketoconazole. No significant pharmacokinetic or pharmacodynamic interactions with almotriptan were observed for fluoxetine or propranolol. Almotriptan clearance was reduced, to a modest degree, by moclobemide and verapamil, which was consistent with the contribution of monoamine oxidase-A and CYP3A4 to the metabolic clearance of almotriptan. Although ketoconazole has a greater effect on almotriptan clearance than verapamil, no dosage adjustment is required when almotriptan is given with these drugs.


The respiratory syncytial virus transcription antiterminator M(2-1) is a highly stable, zinc binding tetramer with strong pH-dependent dissociation and a monomeric unfolding intermediate.
The human respiratory syncytial virus M(2-1) transcription antiterminator is an essential elongation factor required by the RNA polymerase for effective transcription beyond the first two nonstructural genes. Its exclusive presence in pneumovirus among all paramyxovirus suggests a unique function within this small genus. With the aim of understanding its biochemical properties, we investigated this α-helical tetramer by making use of a biophysical approach. We found that the tetramer hydrodynamic radius is considerably extended at high ionic strengths and determined its zinc content to be one atom per monomer. Dissociation-unfolding experiments show a fully reversible and concentration-dependent cooperative transition, but secondary and tertiary structural changes are uncoupled at lower protein concentrations. We detect the presence of a monomeric intermediate, which can be classified as a "late molten globule" with substantial secondary and tertiary structure. Global fittings of experiments from three different probes at two M(2-1) concentrations provide a free energy of dissociation-unfolding of -36.8 ± 0.1 kcal mol(-1), corresponding to a tight dissociation constant of 10(-28) M(3) at pH 7.0. The tetramer affinity is strongly governed by pH, with a free energy change of 13 kcal mol(-1) when pH decreases from 7.0 to 5.0 (K(D) = 10(-18) M(3)). The drastic changes that take place within a pH range compatible with a cellular environment strongly suggest a regulatory effect of pH on M(2-1) structure and biochemical properties, likely affecting transcription and interaction with proteins and RNA.


Insulin resistance and geographical origin: major predictors of liver fibrosis and response to peginterferon and ribavirin in HCV-4.
Hepatitis C virus (HCV) genotype 4 (HCV-4) is increasing in prevalence in Western countries. However, little is known about the severity of the disease and response to treatment. The aim of this study was to assess the predictors (logistic regression) of severe fibrosis (METAVIR score F3-F4), and sustained virological response (SVR) to peginterferon and ribavirin in 226 consecutive HCV-4 patients (Egyptians 40%, Europeans 35% and Africans 24%). Insulin resistance was assessed using the homeostasis model (HOMA-IR). Serum HCV-RNA level (bDNA) and subtypes of HCV (LiPA) were determined for all patients. Insulin resistance (HOMA-IR >3) was present in 105 patients (46%), and was associated with: age >45 years (OR, 2.614; 95% CI, 1.316 to 5.194), body mass index (BMI) >25 kg/m(2) (OR, 2.105; 95% CI, 1.048 to 4.229), serum HCV-RNA >800 000 IU/ml (OR, 3.143; 95% CI, 1.503 to 6.574), severe fibrosis (OR, 2.657; 95% CI, 1.214 to 5.818), and steatosis >30% (OR, 2.488; 95% CI, 1.105 to 5.602). Severe fibrosis was present in 67 patients (29%) and was associated with Egyptian origin (OR, 5.872; 95% CI, 2.747 to 12.553), excessive alcohol intake (OR, 5.311; 95% CI, 1.287 to 21.924), and HOMA-IR >3 (OR, 3.864; 95% CI, 1.859 to 8.034). 108 patients received a 48 week course of peginterferon plus ribavirin. SVR (undetectable serum HCV-RNA (TMA) 24 weeks after treatment stopping) was achieved in 59 patients (55%) and was associated with Egyptian origin (OR, 13.119; 95% CI, 3.089 to 55.706), HOMA-IR <2 (OR, 5.314; 95% CI, 1.953 to 14.459), and non-severe fibrosis (OR, 8.059; 95% CI, 2.512 to 25.855). Insulin resistance and geographical origin are major predictors of liver fibrosis and response to peginterferon and ribavirin in HCV-4 patients. Insulin resistance is frequently encountered in these patients, and correlated independently with serum HCV-RNA.


Magnetic Resonance Imaging Characteristics of Pituitary Abscess: A Review of 51 Cases.
To investigate pituitary abscess (PA) magnetic resonance imaging (MRI) features to improve the neuroradiologic and diagnostic knowledge of this rare disease and guide follow-up treatments. Clinical data were collected for 51 patients with PA, and MRI data were quantitatively reviewed in a retrospective analysis. Clinical factors were analyzed to investigate their relevance. PA neuroimaging showed special radiologic features, including hypointensity or isointensity on T1-weighted imaging (30 patients, 58.8%), isointensity or hyperintensity on T2-weighted imaging (39 patients, 76.5%), and disappearance of the posterior pituitary bright spot in most patients (44 patients, 86.3%). After gadolinium injection, rim or rimlike enhancement was observed in 82.4% of patients (n = 42). Half of those patients showed typical rim enhancement (profound peripheral enhancement with internal hypointensity); others showed atypical rim enhancement with special signs, such as enhanced, thick abscess wall and hyperintense flocculent or cottonlike foci within the internal hypointense region. Almost all patients (96.1%) presented at least 1 sign of adjacent anatomic structure invasion, including peripheral meninges enhancement, pituitary stalk thickening, and paranasal sinus mucosa enhancement. Rank correlation analysis of the clinical time course and MRI characteristics of PA showed a Spearman correlation coefficient of 0.270, indicating borderline significance (P = 0.055). Classic MRI characteristics of PA show T1 signal hypointensity or isointensity, T2 signal isointensity or hyperintensity, and peripheral rim enhancement after gadolinium injection. The MRI appearance of PA may reflect the abscess liquefaction and formation. Despite its profound significance for differential diagnosis, adjacent structure invasion is generally ignored by clinicians.


Controversies in the Management of Vesicoureteral Reflux.
Vesicoureteral reflux (VUR) is the abnormal retrograde flow of urine from the bladder into the upper urinary tract. Diagnosis and subsequent management of VUR have become increasingly controversial, with differing opinions over which children should be evaluated for reflux, and when detected, who should undergo treatment. Management goals include prevention of recurrent febrile urinary tract infection (fUTI) and renal injury while minimizing the morbidity of treatment and follow-up. Management options include observation with or without continuous antibiotic prophylaxis and surgical correction via endoscopic, open or laparoscopic/robotic approaches. Management should be individualized and based on patient age, health, risk of subsequent renal injury, clinical course, renal function, and parental preference.


Recombinant human erythropoietin reduces allogeneic blood transfusion requirements in patients undergoing major orthopedic surgery.
Blood loss is a significant problem encountered in patients undergoing total joint arthroplasty, and is considered to be one of the factors affecting the outcome of the operation. Traditionally these patients have been treated with blood transfusions. The introduction of recombinant human erythropoietin (rHuEpo) into clinical practice enabled assessment of its effectiveness to decrease the allogeneic blood transfusion requirement (BTR), thus avoiding or minimizing transfusion-related complications. Fifteen patients undergoing total hip replacement (THR, 10 patients) and total knee replacement (TKR, 5 patients) in our institute (from January-April 1997), were studied. After signing an informed consent they received daily s.c. rHuEpo (100 IU/kg for those with hemoglobin (Hb) > 13 g/dl, 300 IU/kg for Hb < 13) during the 10 days prior to surgery and the 4 days following the operation. Allogeneic red blood cell (RBC) transfusions were given as needed. Hb levels were measured on days -10, 0, +1.3 and 7 of the procedure and the BTR was recorded. The results were compared with those of previous patients operated on from January-December 1996. Patients who were eligible for the study but refused to participate served as controls. The mean Hb level in the study group prior to rHuEpo administration (day -10) was 13.41 g/dl, similar to those of the control group (13.47 g/dl on day 0). However, the mean Hb levels in the rHuEpo treated patients on days 0, 1, 3 and 7 were 14.37, 11.09, 10.99, and 11.2 g/dl, respectively. This way compared with the levels of 13.47 (p = 0.016), 9.88 (p = 0.024), 9.60 (p = 0.004) and 9.97 g/dl (p = 0.007) in the control patients. The difference between the rHuEpo treated patients and the control patients was more significant among the THR patients than among the TKR patients. Of the 10 rHuEpo-treated THR patients, only a single patient required one allogeneic blood unit, as compared with 23 units transfused to the 30 control patients. None of the rHuEpo-treated TKR patients required blood transfusion as opposed to 4 units needed by the 11 control patients. In total, only one allogeneic blood unit was required by the study group which way calculated to an average consumption of 0.066 blood unit per person, compared with 27 blood units used by the 41 controls, i.e. 0.66 blood units per person (p < 0.001). In the patients treated, rHuEpo was very well tolerated with no adverse effects.


Dementia after ischemic stroke: a population-based study in Rochester, Minnesota (1960-1984)
We used the medical records linkage system for the population of Rochester, Minnesota, to identify persons in the community who had their first cerebral infarct without previous dementia. In this cohort (n = 971), the incidence of dementia in the first year was nine times greater than expected, but if we did not observe dementia in the first year, the risk of dementia in the cohort each year thereafter was about twice the risk in the population. After the first year, a 50% increase was observed in Alzheimer's disease in the cohort compared with that in the community. Although the incidence of dementia increased with increasing age, the standardized morbidity ratios decreased with increasing age. Age, sex (male), and second stroke were significant independent predictors of dementia in a multivariate Cox proportional hazards model. There was no effect of location or clinical severity of infarct on the rate of occurrence of dementia.


[Evaluation of diagnostic accuracy of CRT monitor display for personal computer in the detection of small lung nodules: with particular emphasis on comparison between JPEG and wavelet compression].
To compare observer performance on cathode-ray-tube(CRT) monitors for personal computers with that on conventional radiographs in the detection of small lung nodules. Fifty-eight normal chest radiographs and 58 chest radiographs with a small lung nodule were selected. Ten radiologists examined the original conventional films on a viewbox and digitized (8 bit) uncompressed and compressed images of the same patient on a color CRT monitor with a matrix of 1,600 x 1,200, and rated the presence of lung nodules with a five-level scale of confidence. The methods of compression used in this study were the JPEG and wavelet methods, with compression ratios of 6:1 and 15:1. Results were analyzed by receiver operating characteristic methods. There was no significant difference between film and digitized uncompressed and compressed images obtained by the JPEG and wavelet methods with a compression ratio of 6:1. No statistically significant difference was detected between film and digitized image with wavelet compression at 15:1. However, detection was less accurate on digitized images with JPEG compression at 15:1. Digitized (8 bit) uncompressed and compressed images with a compression ratio of 6:1 are acceptable for the detection of small lung nodules. Digitized compressed images at a compression ratio of 15:1 are also acceptable when the wavelet method is used.


Temporal order and functional analysis of mutations within the Fli-1 and p53 genes during the erythroleukemias induced by F-MuLV.
The erythroleukemias induced by Friend murine leukemia virus (F-MuLV) result from the accumulation of a number of genetic changes, including activation of the Fli-1 proto-oncogene and inactivation of the p53 tumor suppressor gene. We have determined the temporal order of mutation of the genes involved in this multistage malignancy, by serial in vivo transplantation of F-MuLV induced primary erythroleukemias into syngenic Balb/c mice. These primary tumors are capable of growing when transplanted into syngenic mice, but die after several days of in vitro culture. From the transplanted tumors grown in syngenic mice, erythropoietin-dependent cell lines were established in culture that are clonally related to cells in the primary tumors. We show that retroviral insertional activation of the Fli-1 ets family member is the first detectable genetic event in F-MuLV induced primary erythroleukemias. Mutations in the p53 gene were observed in the Epo-dependent cell lines but not in the transplanted erythroleukemias used to establish these cell lines in culture. These data suggest that activation of Fli-1 plays an important role in the early stages of F-MuLV-induced leukemia, perhaps by altering the self-renewal probabilities of erythroid progenitor cells and that p53 mutations immortalize these cells, enabling them to grow in vitro in the presence of Epo.


The association between obesity and oligoclonal band formation in multiple sclerosis patients.
Since some neurological disorders present with increased body-mass index (BMI) and cerebrospinal fluid (CSF) oligoclonal bands (OCB), obesity-induced inflammation has been previously speculated in formation of OCB. We investigated the association between BMI, OCB formation and clinical features of MS in 120 patients with relapsing remitting multiple sclerosis (RRMS), a disease with high OCB positivity incidence. Thirty RRMS patients had BMI≥30 and 100 patients displayed CSF OCB. OCB positive and negative patients had comparable BMI and weight values. Disease duration, annual attack number and EDSS were not correlated with BMI and body weight. Patients with normal and high BMI did not significantly differ by means of OCB positivity, gender, annual attack number, disease duration and EDSS scores. Our results argue against a possible role of obesity in OCB formation. Moreover, obesity does not appear to influence disability and clinical progression of MS patients.


Respiratory health of brickworkers in Cape Town, South Africa. Background, aims and dust exposure determinations.
The respiratory health and work environment of 575 brickworkers in five brickworks in Cape Town, South Africa, were investigated by means of a questionnaire, a physical examination, pulmonary function testing, chest radiography, and dust measurements. This study presents the background, aims, and methods of the determining dust exposure assignments. The mean concentrations of respirable dust and total dust were 2.22 and 15.16 mg/m3, respectively, with a mean free-silica percentage of 2.1%. Subjective and objective, as well as categorical and continuous, dust indicators were constructed for subsequent analyses, together with findings from the health survey, which are reported elsewhere.


[Orthodontic movement through compact bone and spongious bone. The difference in tissue reaction with 2 different forces].
The objective of this study, was to study the contribution of the osseous density factor at the orthodontic movement. For this purpose we compared the histophysiological responses frontally to the roots of the mandibular first molar (at the experimental Wistar rat) according to the direction of displacement (mesio-lingual region). In fact, frontally to the mesial and lingual roots (at the mesio-lingual region) there is compact bony tissue in contrary at the vestibular and distal roots the density is less and there is spongiose osseous tissue. We found the following: the region of pressure is subjected to compression and hyalinisation periodontally from the first day, even with the "ideal" force; the extent of compression and hyalinisation periodontally, reaches the maximum approximately after two days; the extent of compression and hyalinisation periodontally is greater frontally to spongiose osseous tissue and even more greater with a heavy force; the immediate displacement of a tooth is greater frontally to spongiose bone and even more greater with a heavy force; there is bony resorption of the internal alveolarwall adjacent to the hyaline zone, resorption (direct laterally or peripherically) frontally to the compact and to the spongiose osseous tissue; the direct lateral resorption: is slower for the compact bone; frontally to the spongiose bone the lateral direct resorption is associated to an indirect resorption at the endoostic side; the real displacement of a tooth is less rapid in compact bone because the lateral direct resorption is slower and also because of the absence of indirect endoostic resorption; at the time of the real dental displacement, the periodontal space has reestablished; on regard of the spongiose bone, the periodontal space not only reestablishes, but also increases during the real dental displacement; at the level of the periosteum, we can observe a compensatory osseous apposition to the direct lateral bony resorption, only with light forces, on the contrary with heavy forces there is periostic resorption and the tooth movement occurs within the bone (especially frontal to a compact bone); the threshold for stimulating the osteoclasts is lower than the ideal force frontal to the compact bone. On the contrary, frontal to the spongiose bone, the threshold for stimulating the osteoclasts, is situated higher to the ideal force.


Intestinal bypass surgery produces conditioned taste aversion in rats.
Intestinal bypass surgery reduced the body weight of obese humans and rats primarily by decreasing food consumption. The present study reports that jejunoileal bypass surgery in rats produces a strong and persistent conditioned taste aversion to a novel-flavored solution with which it is paired. This finding indicates that the jejunioleal bypass produces a long-lasting malaise, which may be an important factor in the anorexia and weight loss produced by the operation.


Free tissue transfer of the rectus abdominis myoperitoneal flap for oral reconstruction in a dog.
A five-month-old intact/male Boxer dog was presented 5-days following bite wound trauma to the maxillary region resulting in an oronasal fistula extending from the maxillary canine teeth to the soft palate. Multiple surgical procedures using local, buccal mucosal flaps failed to repair the oronasal fistula. Free tissue transfer of the rectus abdominis myoperitoneal flap using microvascular surgical techniques was successful in providing soft tissue reconstruction of the hard palate area. Complications of these surgical techniques included muscle contraction and subsequent muzzle distortion. Small, refractory oronasal fistulae at the perimeter of the myoperitoneal flap were repaired by primary wound closure.


Characterization and behavior of composite hydrogel prepared from bamboo shoot cellulose and β-cyclodextrin.
Carboxymethyl cellulose was derived from bamboo shoot cellulose via chemical modification and was prepared into composite hydrogels by cross-linkage with β-cyclodextrin using epichlorohydrin as crossing agent. The structure of the prepared hydrogel was characterized by Fourier transform infrared spectroscopy, thermogravimetric analysis and scanning electron microscopy. The results showed that the prepared composite hydrogel was sensitive to surrounding changes in pH value, temperature and ionic strength. Under the surroundings of low temperatures and high pH values, the prepared hydrogel had significant high swelling ratios (23338±988% at 15°C and 6937±112% at pH 8.0, respectively). In the solution of 0.1mol/L NaCl, the hydrogel showed the maximum water retention rate (48.73%). Sodium salicylate was used as the model drug to study the behaviors of hydrogel adsorption and release in simulated intestinal (at pH 7.4) and gastric liquid (at pH 1.8) surroundings. The prepared composite hydrogel exhibited higher drug release ratio in simulated intestinal liquid (63.09% after 380min) than in gastric liquid (22.09% after 400min). These pH responses of the prepared composite hydrogel showed its potential applications, especially as the drug carrier to attain control release of drugs under different surrounding conditions or organs in human body.


Evaluation of an ion-exchange membrane for the purification of plasmid DNA.
Separation and purification of large quantities of plasmid DNA (pDNA) is a particularly difficult manufacturing issue because of the relatively low capacity, flow rate and purity observed using traditional bead-based chromatography. The objective of the present study was to evaluate the performance of anion-exchange membranes for the purification of pDNA from Escherichia coli lysate solution. The fate of host-cell protein and endotoxin relative to pDNA was measured and used to calculate recoveries, mass balances, dynamic capacities and purification factors as a function of the flow rate and loading volume of the lysate solution. Breakthrough curves were not sigmoidal and symmetric in shape. They rose sharply at first, and then slowly towards, but never reaching, saturation. Conversely, elution curves were independent of flow rate. pDNA bound tightly to the membranes, whereas protein and endotoxin did not. Dynamic binding capacity for pDNA was 20-25 times greater, and the flow rate was 55-550 times greater, than values observed for beads. However, some pDNA bound irreversibly to the membrane surface and was not removed completely during elution. The intrinsic rate of pDNA adsorption to the membrane was found to be rate-limiting, whereas effects of liquid-phase mass transfer and flow non-idealities were negligible. These results were interpreted using models of adsorption that included steric effects using the 'car-parking-problem' model, and surface residence time effects using the spreading model. This work demonstrated the advantages of ion-exchange membranes compared with beads for the purification of large biomolecules such as pDNA.


αRep A3: A Versatile Artificial Scaffold for Metalloenzyme Design.
αRep refers to a new family of artificial proteins based on a thermostable α-helical repeated motif. One of its members, αRep A3, forms a stable homo-dimer with a wide cleft that is able to accommodate metal complexes and thus appears to be suitable for generating new artificial biocatalysts. Based on the crystal structure of αRep A3, two positions (F119 and Y26) were chosen, and independently changed into cysteine residues. A phenanthroline ligand was covalently attached to the unique cysteine residue of each protein variant, and the corresponding biohybrids were purified and characterized. Once mutated and coupled to phenanthroline, the protein remained folded and dimeric. Copper(II) was specifically bound by the two biohybrids with two different binding modes. Furthermore, the holo-biohybrid A3F119NPH was found to be capable of enantioselectively catalyzing Diels-Alder (D-A) cycloadditions with up to 62 % ee. This study validates the choice of the αRep A3 dimer as a protein scaffold and provides a promising new route for the design and production of new enantioselective biohybrids based on entirely artificial proteins obtained from a highly diverse library.


Alloimmune anti-H-2 mouse sera inhibit human mixed lymphocyte reactions.
The inhibitory effect of alloimmune congenic mouse anti-H-2f serum on human mixed lymphocyte cultures was investigated. The anti-H-2 serum tested produced a strong inhibitory effect on human allogeneic MLR. This effect was more pronounced when HLA-A2 positive cells were involved in the MLR. More experiments are needed to substantiate the finding that both stimulator and responder functions of the human lymphocytes are impaired. The anti-H-2 serum also inhibited the human lymphocyte response to PHA.


Nanomechanical contrasts of gel and fluid phase supported lipid bilayers.
Lipid bilayers exhibit structural diversity that contributes to the complex properties of the cell membrane. We use interfacial force microscopy to correlate mechanical properties with the two-dimensional phase behavior of supported lipid bilayers (SLBs). Upon indentation by a 500 nm tungsten tip, a contrast in the mechanical response is observed for gel vs fluid phase SLBs. We measure the yield force and time scale for recovery for these films. Consistent with a gel phase, a DSPC SLB has a relatively high yield force and slow recovery. In the higher mobility fluid phase, a DLPC SLB has a lower yield force and completely recovers within the experimental time scale. Friction measurements offer further contrast between the two SLBs.


Interference of heparin in carcinoembryonic antigen radioimmunoassays.
A false Roche carcinoembryonic antigen (CEA) activity could be detected in all commercial and noncommercial heparin preparations we examined. The possibility of 'due to contamination' has been ruled out. Using the Roche procedure, heparin solutions, in the absence of CEA, gave positive CEA activity; on the other hand, no CEA activity was detected in solutions containing only heparin when the Abbott Kit was used. When heparin was present in specimens containing CEA, the Abbott Kit underestimated the CEA activity, whereas the Roche Kit gave false elevated values. However, the negative effect of heparin could be reduced by heat treatment in the presence of plasma proteins.


Phase II study of bendamustine combined with rituximab in relapsed/refractory mantle cell lymphoma: efficacy, tolerability, and safety findings.
In most cases of relapsed/refractory mantle cell lymphoma (MCL), patients respond to salvage therapy, though typically responses are partial and/or transient followed by disease progression, even with newer agents (e.g., ibrutinib). In this multicenter, open-label, single-arm, phase II study, patients with relapsed/refractory non-blastoid MCL received bendamustine 90 mg/m(2) (days 1 and 2) and rituximab 375 mg/m(2) (day 1) for 6 planned 28-day cycles. Functional imaging with 18-fluoro-2-deoxy-D-glucose positron emission tomography/computed tomography (18F-FDG PET/CT) was conducted at baseline and after cycle 6. Forty-five patients were enrolled (median age, 70 years; 82 % stage IV disease; median number of prior chemotherapies, 2 [range, 1-4]), showing an overall response rate (ORR; primary efficacy measure) of 82 % (complete response [CR], 40 %; partial response, 42 %). In the 32 patients with complete 18F-FDG PET/CT data, 75 % achieved a complete metabolic response. Median duration of response was 1.6 years, 1-year progression-free survival was 67 %, and 3-year overall survival was 55 %. Main non-hematologic adverse events were nausea (69 %), fatigue (56 %), decreased appetite (42 %), constipation (38 %), diarrhea (36 %), vomiting (36 %), and decreased weight (31 %). Grade 3/4 neutropenia and lymphopenia occurred in 44 and 89 % of patients, respectively. ORR and CR rate compared favorably with single-agent ibrutinib (ORR, 67 %; CR, 23 %); bendamustine-rituximab is an effective therapy with manageable toxicity in relapsed/refractory MCL.


Guidelines for the safe use of dental and maxillofacial CBCT: a review with recommendations for South Africa.
The aim of this manuscript is to provide the South African dental profession with guidelines and recommendations on the safe and responsible clinical use of cone beam computed tomography (CBCT) equipment. Dental practitioners who prescribe and refer patients for CBCT examinations as well as those who operate CBCT systems and interpret the images, share responsibility for the use of CBCT systems. Applicable information pertaining to the current status of the use of CBCT was reviewed, and the Directorate of Radiation Control of the Department of Health, South Africa, was invited to participate in the development of evidence-based guidelines on the safe use of cone beam computed tomography in dentistry. These guidelines establish a basis for discussions that are likely to lead to legislation.


Expression of RNF8 on cochlear apoptosis and aging in mice of different ages.
This study aimed to investigate the role of RNF8 (RING finger protein 8) in DNA damage repair in mice of different ages, and to provide new insight into the pathology and treatment of senile deafness. Sixteen C57BL/6J mice aged 8 weeks, 16 weeks and 32 weeks were obtained by paired reproduction. The mice of three age groups were equally divided into two groups, named experimental group (RNF8 gene knockout) and control group (no knockout). The cochlear hair cells, stria vascularis and spiral ganglion cells were observed by HE (hematoxylin-eosin) staining. The degree of DNA damage and the related expressions were observed by immunofluorescence γ-H2AX staining and 8-OH immunohistochemical staining, and the aging of damaged cells was detected by lipofuscin and β-galactosidase staining. HE staining showed that the changes of cochlear hair cells, stria vascularis and spiral ganglion cells were obvious in the same group of mice at different ages. Compared with the control group, the aging changes of cochlear hair cells, stria vascularis and spiral ganglion cells were more significant in the experimental group. Immunofluorescence γ-H2AX staining showed H2AX phosphorylation in injured cells. The aging of cochlea in mice changed, and staining of β-galactosidase in the experimental group suggested that the striae of blood vessels were changed with age at 32 weeks old and staining of lipofuscin showed dark brown staining around the nucleus (P < 0.05). In conclusion, the deletion of RNF8 is an important cause of morphological changes in the cochlea of mice. The deletion of RNF8 accelerates the aging of the cochlea of mice, suggesting that the apoptosis of the cochlea could contribute to aging in RNF8 gene-deficient mice.


N-oxygenation of stobadine, a gamma-carboline antiarrhythmic and cardioprotective agent: the role of flavin-containing monooxygenase.
1. N-oxygenation of stobadine, a gamma-carboline antiarrhythmic and cardioprotective agent, was investigated in vitro using rat liver preparations. 2. Stobadine N-oxygenase activity was located mainly in the microsomal fraction and exhibited a requirement for oxygen and NADPH. The apparent Km and Vmax values for the process were, respectively, 350 microM and 3.48 nmol/mg protein per min. 3. N-oxygenation of stobadine in rat liver microsomes was not affected by SKF 525-A, carbon monoxide, metyrapone, cyanide or n-octylamine. When guinea-pig liver microsomes were used the reaction was activated by n-octylamine. 4. N-oxygenase activity was strongly inhibited by methimazole and depressed by phenobarbital and 3-methylcholantrene pretreatment. 5. The above results, along with the pH optimum of 8.4, strongly indicate the involvement of flavin-containing monooxygenase in the metabolic N-oxygenation of stobadine. 6. Species difference in liver microsomal N-oxygenase activity was evident, the order of activity being guinea-pig greater than rat greater than rabbit.


Patterns of autoantibody expression in pediatric and adult systemic lupus erythematosus.
To characterize patterns of autoantibody expression in pediatric and adult systemic lupus erythematosus (SLE), IgG and IgM antibodies to cardiolipin (aCL) and single-stranded DNA (anti-DNA) were measured in sera from 32 pediatric and 36 adult patients. Antibody levels were assessed by isotype-specific ELISA and compared with 15 pediatric disease controls and 36 healthy adult controls. These determinations revealed significant differences in the pattern of IgG and IgM autoantibody expression in adult and pediatric patients. IgM aCL were more common in the pediatric than adult SLE population with 11 of 32 pediatric patients being positive compared to only 4 of 36 in the adult group (p less than 0.05). Conversely, enhanced IgG autoantibody production was observed in the adult group as 27 of 36 adult patients were positive for IgG anti-DNA compared to only 16 of 32 children (p less than 0.05). Analysis of sequential sera showed differences in the temporal expression of IgG and IgM autoantibodies as well as variability between anti-DNA and aCL responses over time. Our data suggest differences in the isotype profile of autoantibody responses in pediatric and adult SLE and provide further evidence for heterogeneity in the mechanisms of autoantibody production.


Procedural sedation and analgesia in a Canadian ED: a time-in-motion study.
Some patients presenting to emergency departments (EDs) suffer from conditions requiring potentially painful treatment; procedural sedation and analgesia (PSA) are important components of their management. The purpose of this study was to determine the resources used during the administration of PSA. This prospective observational study was conducted in a Canadian urban teaching center. Detailed data concerning the dosage of PSA medications, adverse events, and ED times for patients requiring PSA for treatment of fractures, reductions of joint dislocations, and cardioversion for atrial fibrillation were collected. Descriptive analyses included proportions, means with standard deviations, and medians with interquartile ranges. Of the 177 PSA cases considered for analysis, 69.5% were orthopedic manipulations and 30.5% were cardioversions. Propofol alone or combined with fentanyl was the commonest medication, and 27 minor adverse events were documented. The median number of staff used in each PSA was 4 (4, 4). The median time from triage to the start of the procedure was 175 minutes (98, 259). The median time from the end of monitoring to discharge was 186 minutes (104, 316). The median time from the start of PSA administration to the end of patient monitoring was 12 minutes for fractures/dislocations and 7 minutes for cardioversion. The total ED length of stay was 6.6 hours. Procedural sedation and analgesia are potentially time-consuming interventions requiring the coordination of ED staff; delays in procedures represent opportunities to reduce ED overcrowding. Procedural sedation and analgesia guidelines may assist with standardization.


The use of X-ray fluorescence (XRF) analysis in predicting the alkaline hydrothermal conversion of fly ash precipitates into zeolites.
The use and application of synthetic zeolites for ion exchange, adsorption and catalysis has shown enormous potential in industry. In this study, X-ray fluorescence (XRF) analysis was used to determine Si and Al in fly ash (FA) precipitates. The Si and Al contents of the fly ash precipitates were used as indices for the alkaline hydrothermal conversion of the fly ash compounds into zeolites. Precipitates were collected by using a co-disposal reaction wherein fly ash is reacted with acid mine drainage (AMD). These co-disposal precipitates were then analysed by XRF spectrometry for quantitative determination of SiO(2) and Al(2)O(3). The [SiO(2)]/[Al(2)O(3)] ratio obtained in the precipitates range from 1.4 to 2.5. The [SiO(2)]/[Al(2)O(3)] ratio was used to predict whether the fly ash precipitates could successfully be converted to faujasite zeolitic material by the synthetic method of [J. Haz. Mat. B 77 (2000) 123]. If the [SiO(2)]/[Al(2)O(3)] ratio is higher than 1.5 in the fly ash precipitates, it favours the formation of faujasite. The zeolite synthesis included an alkaline hydrothermal conversion of the co-disposal precipitates, followed by aging for 8h and crystallization at 100 degrees C. Different factors were investigated during the synthesis of zeolite to ascertain their influence on the end product. The factors included the amount of water in the starting material, composition of fly ash related starting material and the FA:NaOH ratio used for fusing the starting material. The mineralogical and physical analysis of the zeolitic material produced was performed by X-ray diffraction (XRD) and nitrogen Brunauer-Emmett-Teller (N(2) BET) surface analysis. Scanning electron microscopy (SEM) was used to determine the morphology of the zeolites, while inductively coupled mass spectrometry (ICP-MS), Fourier transformed infrared spectrometry (FT-IR) and Cation exchange capacity (CEC) [Report to Water Research Commission, RSA (2003) 15] techniques were used for chemical characterisation. The heavy and trace metal concentrations of the zeolite products were compared to that of the post-synthesis filtrate and of the precipitate materials used as Si and Al feed stock for zeolite formation, in order to determine the trends (increase or decrease) and ultimate fate of any toxic metals incorporated in the co-disposed precipitated residues.


Intermittent Cooling During Judo Training in a Warm/Humid Environment Reduces Autonomic and Hormonal Impact.
Carballeira, E, Morales, J, Fukuda, DH, Granada, ML, Carratalá-Deval, V, López Díaz de Durana, A, and Stout, JR. Intermittent cooling during Judo training in a warm/humid environment reduces autonomic and hormonal impact. J Strength Cond Res 33(8): 2241-2250, 2019-The purpose of this study was to identify the effects of superficial cooling on physiological responses while training in a warm, humid environment during an international Judo training camp. Sixteen judokas (8 women and 8 men) participated in the experiment. Four high-level women and 4 men were randomly assigned to wear a cooling vest (vest group [VG]) during the recovery periods within a training session (i.e., 8 bouts of 5-minute fighting with 5-minute rest) and up to 10 minutes after the session, whereas the remaining athletes in the control group (CG) trained without the use of any cooling aids. No differences between groups were reported in well-being before the session or in perceived fatigue after the session. The temperature was increased after the training session (p = 0.02) without significant differences between groups; however, CG demonstrated a moderate effect size (ES = 0.95, 90% confidence interval [CI] = 0.09-1.82; probability of superiority [PS] = 74.9%) in contrast to the small effect for VG (ES = 0.28, 90% CI = -0.55 to 1.11; PS = 57.9%). There were time × group interactions for heart rate variability (lnRMSSD) (p = 0.006; VG vs. CG, PS = 79.0%) and the dehydroepiandrosterone-cortisol ratio (DHEA/C ratio) (p = 0.04; VG vs. CG, PS = 99.9%). Vest group preserved the cardiac autonomic control (p > = 0.05; ES = -0.06, 90% CI = -0.88 to 0.76; PS = 51.7%) compared with the large decrement of CG (p < 0.05; ES = -1.18, 90% CI = -2.07 to -0.29; PS = 74.9%). Furthermore, VG showed an increase of DHEA/C (p = 0.002) from presession to postsession based on a moderate decrease of cortisol (p > = 0.05; ES = -0.67, 90% CI = -1.52 to 0.17; PS = 68.2%) with a concomitant small increase of DHEA (p > = 0.05; ES = 0.46, 90% CI = -0.38 to 1.29; PS = 62.7%). Conversely, the CG showed a moderate effect for increased DHEA and a small effect for increased cortisol after training. No significant interactions or main effects were shown for isometric handgrip values. Cooling vests diminished the cardiovascular strain and hormonal impact of the Judo training session in high-level athletes and may be considered for recovery purposes during exercise in warm/humid environments.


Hypothyroidism in elderly hospital patients.
Three hundred and ninety-six patients aged 65 years and over who were admitted to Waitakere Hospital in 1967-77 were screened for hypothyroidism using an adjusted serum thyroxine, serum thyrotrophin, or both. Nine new cases were found. Eleven other patients were already under treatment with thyroxine, though one had defaulted; in six of these the validity of the diagnosis was confirmed. A further five patients had marginally raised levels of serum thyrotrophin, of doubtful significance. There were therefore 15 proven cases of hypothyroidism, an incidence of 3.8%.


The effects of ethanol ingestion on the accuracy of pulmonary diffusing capacity measurement.
Erroneous diffusing capacity of the lung for carbon monoxide (Dlco) values as measured by spectrophotometry were observed at our facility in ethanol-intoxicated subjects. An atypical methane curve tracing was noted in these subjects. We hypothesized that ethanol intoxication interferes with Dlco measurements obtained using methane and designed a study to assess the blood ethanol level at which this occurs. Dlco and breath ethanol levels were measured at baseline and after escalating doses of ethanol in seven healthy subjects. Pulmonary function testing laboratory of a university hospital. Seven healthy volunteers. Dlco measurement at baseline and after escalating doses of ethanol. We found no significant change in measured Dlco values for a wide range of blood ethanol levels (from 0.006 to 0.12 mg/dL). However, subsequently, an abnormal methane curve and Dlco were again observed in an intoxicated subject whose ethanol blood level was 0.22 mg/dL. We conclude that interference between breath ethanol level and Dlco measurement exists only for blood ethanol levels well above the legal limit for intoxication. Based on our observations, we suggest that ethanol intoxication should be suspected when an abnormal Dlco measurement occurs along with an abnormal methane curve tracing. If intoxication is confirmed, precautions should be taken and referral for treatment of possible ethanol dependence should be offered.


Active and passive behaviour in the regulation of stiffness of the lateral wall in outer hair cells of the guinea-pig.
The stiffness of the outer hair cell (OHC) lateral wall, measured by the micropipette aspiration technique, is non-linear, decreasing from the ciliary pole (stiffness parameter Sp 1.83+/-0.13 nN/microm n=10) towards the cell base (Sp 1.14+/-0.16 nN/microm, n=10) irrespective of the cochleoapical or cochleobasal origin of the cells. The length of the aspirated lateral wall segment was related exponentially to the duration of the applied negative pressure (6 cm H2O) in the synaptic region of the OHCs whereas an active, sigmoid component was observed between 30 and 60 s in the supranuclear regions. A significant increase of the midlateral wall stiffness (to 1.91+/-0.23 nN/microm; n=10) was observed in calcium-free medium and the sigmoid component of the response of the lateral wall was abolished. Salicylate (5 mM) had no significant effect on the active sigmoid behaviour of the lateral wall (n=10). Gadolinium (5 mM), a non-specific cation channel blocker, increased the stiffness of the lateral wall and attenuated the active component (n=10). The motor protein prestin thus does not seem to be involved in the active stiffness regulation seen in this study. A role for the cortical cytoskeleton in the regulation of stiffness seems reasonable according to our model. The mechanism may involve calcium-dependent metabolic modification of cytoskeletal or membrane proteins.


[1-H2N-CB11F11]---synthesis and reactions of a functionalized fluorinated carbadodecaborate anion.
In anhydrous hydrogen fluoride, K[1-H2N-CB11H11] is fluorinated with elemental fluorine to produce K[1-H2N-CB11F11]. Under strong alkaline conditions, two fluorine atoms of the [1-H2N-CB11F11]- anion are regioselectively exchanged, yielding the [1-H2N-4,6-(HO)2-CB11F9]- anion via [1-H2N-6-HO-CB11F10]- as an intermediate. Both hydroxycarborate anions were isolated as [Ph4P]+ salts. All of the species were characterized by IR, Raman, and multi-NMR spectroscopy, thermal analysis (DSC) as well as by mass spectrometry (MALDI). The assignment of the NMR signals was supported by DFT calculations. Solid-state structures of K[1-H2N-CB11F11], [BzPh3P][1-H2N-CB11F11], [Ph4P][1-H2N-4,6-(HO)2-CB11F9], [Ph4P][1-H2N-6-HO-CB11F10], and [BzPh3P][1-H2N-CB11H11] were determined by single-crystal X-ray diffraction.


The effect of zinc tape upon wound healing. A biochemical, histochemical and histological study in rats.
The healing of excisional wounds in rats, which were treated with zinc tape, gauze sponge or porcine skin, was studied. Wound closure was completed earlier in zinc-tape-treated wounds than in wounds treated with a gauze sponge or porcine skin. Wound contraction was more pronounced in gauze- and porcine skin-treated wounds than in zinc-tape-treated wounds. More foreign body giant cells were seen 14 days after operation in gauze-treated contra zinc tape-treated wounds, whereas hydroxyproline concentration was higher in tape-treated wounds. Histochemically the acid phosphatase activity was pronounced in macrophages, foreign body giant cells and fibroblasts. The alkaline phosphatase activity was pronounced in granulocytes and fibroblasts. Quantitatively a reduction in alkaline phosphatase activity was seen in gauze-treated wounds from 7 to 14 days. A decrease was seen in albumin concentration in gauze- as well as zinc-tape-treated animals which was most pronounced after 7 days in the gauze-treated animals.


Design of a semi-implantable hearing device for direct acoustic cochlear stimulation.
A new hearing therapy based on direct acoustic cochlear stimulation was developed for the treatment of severe to profound mixed hearing loss. The device efficacy was validated in an initial clinical trial with four patients. This semi-implantable investigational device consists of an externally worn audio processor, a percutaneous connector, and an implantable microactuator. The actuator is placed in the mastoid bone, right behind the external auditory canal. It generates vibrations that are directly coupled to the inner ear fluids and that, therefore, bypass the external and the middle ear. The system is able to provide an equivalent sound pressure level of 125 dB over the frequency range between 125 and 8000 Hz. The hermetically sealed actuator is designed to provide maximal output power by keeping its dimensions small enough to enable implantation. A network model is used to simulate the dynamic characteristics of the actuator to adjust its transfer function to the characteristics of the middle ear. The geometry of the different actuator components is optimized using finite-element modeling.


Comparative quantitative proteomics to investigate the remodeling of bioenergetic pathways under iron deficiency in Chlamydomonas reinhardtii.
The basic question addressed in this study is how energy metabolism is adjusted to cope with iron deficiency in Chlamydomonas reinhardtii. To investigate the impact of iron deficiency on bioenergetic pathways, comparative proteomics was combined with spectroscopic as well as voltametric oxygen measurements to assess protein dynamics linked to functional properties of respiratory and photosynthetic machineries. Although photosynthetic electron transfer is largely compromised under iron deficiency, our quantitative and spectroscopic data revealed that the functional antenna size of photosystem II (PSII) significantly increased. Concomitantly, stress-related chloroplast polypeptides, like 2-cys peroxiredoxin and a stress-inducible light-harvesting protein, LhcSR3, as well as a novel light-harvesting protein and several proteins of unknown function were induced under iron-deprivation. Respiratory oxygen consumption did not decrease and accordingly, polypeptides of respiratory complexes, harboring numerous iron-sulfur clusters, were only slightly diminished or even increased under low iron. Consequently, iron-deprivation induces a transition from photoheterotrophic to primarily heterotrophic metabolism, indicating that a hierarchy for iron allocations within organelles of a single cell exists that is closely linked with the metabolic state of the cell.


Molecular mechanism of YbtA-mediated transcriptional regulation of divergent overlapping promoters ybtA and irp6 of Yersinia enterocolitica.
Genes of Yersiniae spp. involved in production of the siderophore yersiniabactin are located on the high-pathogenicity island (HPI). Their transcription is controlled by the AraC/XilS-like transcriptional regulator YbtA encoded within the HPI. YbtA-regulated divergent and overlapping ybtA and irp6 promoters contain three YbtA binding sites, RS1, RS2 and RS3. Deleting RSs systematically and using ybtA and irp6 transcriptional fusions, we determined that different modes of YbtA binding are responsible for activation of irp6 and repression of ybtA. Based on these data, we propose a model of irp6 and ybtA promoter regulation.


Clinical and demographic factors and outcome of amyotrophic lateral sclerosis in relation to population ancestral origin.
To review how the phenotype and outcome of amyotrophic lateral sclerosis (ALS) change with variations in population ancestral origin (PAO). Knowledge of how PAO modifies ALS phenotype may provide important insight into the risk factors and pathogenic mechanisms of the disease. We performed a systematic review and meta-analysis of the literature concerning differences in phenotype and outcome of ALS that relate to PAO. A review of 3111 records identified 78 population-based studies. The 40 that were included covered 40 geographical areas in 10 subcontinents. Around 12,700 ALS cases were considered. The results highlight the phenotypic heterogeneity of ALS at time of onset [age, sex ratio (SR), bulbar onset], age at diagnosis, occurrence of comorbidities in the first year after diagnosis, and outcome (survival). Subcontinent is a major explanatory factor for the variability of the ALS phenotype in population-based studies. Some markers of ALS phenotype were homogeneously distributed in western countries (SR, mean age at onset/diagnosis) but their distributions in other subcontinents were remarkably different. Other markers presented variations in European subcontinents (familial ALS, bulbar onset) and in other continents. As a consequence, ALS outcome strongly varied, with a median survival time from onset ranging from 24 months (Northern Europe) to 48 months (Central Asia). This review sets the scene for a collaborative study involving a wide international consortium to investigate, using a standard methodology, the link between ancestry, environment, and ALS phenotype.


Potentiation of Ca(2+)-activated secretory activity by a cAMP-mediated mechanism in avian salt gland cells.
In the avian salt gland carbachol (CCh) evokes oscillations in K+ and Cl- current that are sufficient to fully activate secretory activity. Employing the perforated patch-clamp technique, we demonstrate that beta-adrenergic receptor activation stimulates a sustained adenosine 3',5'-cyclic monophosphate (cAMP)-dependent Cl- current with no increase in K+ current. This evokes only a modest increase in secretory activity. However, application of isoproterenol in the presence of a threshold dose of CCh results in maximal secretory activity. Membrane potential measurements demonstrate that isoproterenol stimulates a sustained membrane depolarization from approximately -45 mV to the Cl- equilibrium potential (ECl), whereas CCh evokes oscillations in membrane potential to levels more negative than ECl, representing a mixture of K+ and Cl- conductances. We conclude that, in agreement with current models of fluid secretion, maximal stimulation can only be achieved with simultaneous activation of both K+ and Cl- currents. Because isoproterenol fails to stimulate a K+ current, Cl- secretion is reduced as the driving force for Cl- secretion is dissipated. However, if a driving force is imposed by increasing K+ channel activity (by coadministering CCh), Cl- efflux is sustained. These results could provide a basis for the marked potentiation of Ca(2+)-mediated secretion by agonists that increase cAMP seen in in vivo studies of salivary glands and other exocrine tissues.


Duloxetine: new indication. Depression and diabetic neuropathy: too many adverse effects.
(1) Several classes of antidepressants are available. The main difference between these classes is in their short-term pharmacological effects, leading to different patterns of adverse effects. Some antidepressants, especially tricyclics, have positive risk-benefit balances in the treatment of diabetic neuropathy. (2) Duloxetine, a compound chemically related to fluoxetine, appears to have a short-term mechanism of action similar to that of venlafaxine. In the European Union, duloxetine was first approved for female urinary stress incontinence. Another brand of duloxetine has since been marketed for depression and neuropathic pain in diabetic patients. (3) Duloxetine at a dose of 60 mg once a day showed moderate efficacy in 2 placebo-controlled trials. At this dose, however, there are no other comparative trials. It is therefore not possible to know whether duloxetine is as effective as other antidepressants. (4) Two placebo-controlled trials involving patients with pain due to diabetic neuropathy concluded that a dose of 60 mg/day had efficacy, although of doubtful clinical relevance. In the absence of comparative trials, however, we do not know if this efficacy is even equivalent to that of a tricyclic antidepressant used as an analgesic. (5) In fibromyalgia, a controversial clinical diagnosis, two double-blind placebo-controlled trials involving 207 and 354 patients failed to prove that duloxetine had tangible analgesic efficacy. It is therefore appropriate that this use is not mentioned in the "Indications" section of the summary of product characteristics (SPC). (6) The assessment of duloxetine in depression and neuropathic pain confirms existing data on its gastrointestinal, neuropsychological and hepatic adverse effects. In these trials, duloxetine increased blood pressure in a dose-dependent manner. (7) Duloxetine is metabolized by cytochrome P450 isoenzymes CYP 1A2 and CYP 2D6, creating an important risk of interactions with other drugs. (8) In practice, duloxetine currently has no place in the treatment of depression or diabetic neuropathy. Its efficacy has not yet been demonstrated to be even equivalent to that of other available drugs, and it has too many adverse effects, given this degree of uncertainty.


Public and personal depression stigma in a rural American female sample.
We examined public and personal stigma among a community sample of 1,000 women living in primarily rural counties of Western Kentucky. Data on demographics, depression, stigma, health information sources, and availability of health services were collected via a random digit dial survey. The prevalence of depression was 15.7%. The majority of respondents (82.2%) reported congruent levels of stigma with 11.6% reporting high public and high personal stigma. However, 17.8% of respondents reported incongruent public and personal stigma. The 7.5% of women with low public and high personal stigma were older and less educated, preferred anonymous sources of health information, and reported better availability of health services. The 10.3% of women with high public and low personal stigma were younger and more educated, preferred interpersonal sources of health information, and reported poorer availability of health services. In multivariate analyses, depression and lower education were associated with any incongruent stigma, while rural residence and White race/ethnicity was associated with high personal and public stigma. Psychiatric nurses should develop community-based and targeted, point-of-care interventions to reduce public and personal stigma among rural women.


The use of chitosan to increase the stability of calcium alginate beads with entrapped yeast cells.
A method was developed for increasing the stability of calcium alginate beads with entrapped Saccharomyces cerevisiae cells. Polyelectrolyte complexes were used as surface-coating materials by dropping alginate into chitosan. The coacervation agent sodium triphosphate was needed for the formation of the polyelectrolyte complexes. Gel beads maintained their rigidity during ethanol production and were chemically stable in phosphate buffer. No differences in ethanol production were found between S. cerevisiae immobilized in calcium alginate and in calcium alginate coated with polyelectrolyte complexes. The maximum productivity by immobilized S. cerevisiae cells was 74.1 g of ethanol/h per g of gel beads obtained at pH 4.8-6.0 and at 30 degrees C.


[Infected aneurysms of the aorta].
Mycotic aneurysms, a quite rare syndrome, are classified into three groups: primary (the most frequent one), secondary to bacterial endocarditis, and secondary to direct inoculation (infections focus or trauma). Primary aneurysms are related to bacteremia with bacterial setting of an atheromatous lesion, most often. Clinical presentation is often misleading or delayed from infectious process. However, presence of clinical aneurysm, pain, and infectious syndrome should suggest diagnosis and lead to immediate antibiotic therapy and rapid surgical management, as rupture of the aneurysm is a frequent, serious and ineluctable complication. Importance of microbiologic search is to be emphasized as the diversity of the responsible microorganisms is very high; waiting for bacteriological results, a bactericidal double antibiotherapy should be instituted.


Sulforaphane induces cell cycle arrest and apoptosis in murine osteosarcoma cells in vitro and inhibits tumor growth in vivo.
Sulforaphane (SFN), a naturally occurring isothiocyanate, is an attractive agent due to its potent anticancer effects. SFN suppresses the proliferation of various cancer cells in vitro and in vivo. In this study, we report that SFN inhibited the proliferation of cultured murine osteosarcoma LM8 cells. Twenty micromolar SFN completely inhibited the growth of LM8 cells and caused G2/M-phase arrest. SFN induced the expression of p21(WAF1/CIP1) protein causing the cell cycle arrest in a dose-dependent manner. SFN induced apoptosis which was characterized by the appearance of cells with sub-G1 DNA content and the cleavage and activation of caspase-3. We showed that SFN induced the growth arrest and up-regulated the expression of p21(WAF1/CIP1) protein in a p53-independent manner in human osteosarcoma MG63 cells. We found that intraperitoneal administration of SFN (1 or 2 mg, 5 times/week) significantly inhibited the growth of LM8 xenografts to <30% of the controls in a preclinical animal model without causing any toxicity. In osteosarcoma cells, our findings provide in vivo evidence for the efficacy of SFN against the advanced growth of tumor. We showed that SFN induces cell cycle arrest and apoptosis in osteosarcoma cells and inhibits tumor xenograft growth. Furthermore, SFN is a potent inducer of p21(WAF1/CIP1) in osteosarcoma cells. These results raise the possibility that SFN may be a promising candidate for molecular-targeting chemotherapy against osteosarcoma.


Microcosting method for small-volume injectables.
The costs of preparing different small-volume injectables (SVIs) in a centralized i.v. admixture pharmacy in a 650-bed teaching hospital were calculated using a microcosting method. The types of SVIs produced and methods of preparation were identified. Time-and-motion and work-sampling studies were performed to determine the amount of pharmacist and technician time spent in SVI production, auxiliary activities, and nonproductive activities. The costs of material and equipment used were calculated, and waste-cost factors based on percentages of wasted SVIs were determined. To determine the final cost per unit prepared, the calculated costs were added and multiplied by the appropriate waste factors. On the average, it took 5.57 minutes of total personnel time to prepare an SVI. It took an average of 192.4 seconds and 16.6 seconds for the technicians and pharmacists, respectively, to reconstitute a vial of powder and prepare an SVI in a minibag. Bulk prepacked items and admixtures prepared from vials of powder for reconstitution and stored in minibags were, respectively, the least and most expensive SVIs. By using microcosting methods to determine the cost of preparing SVIs, this institution discovered that the cost figures were much higher than it had anticipated. These data can be used to improve productivity and cost efficiency and to compare the costs of in-house and commercially available preparations.


Histologic correlates of metastasis in primary invasive squamous cell carcinoma of the lip.
Clinical and histologic features of 44 invasive squamous cell carcinomas of the lip treated by surgical excision were correlated with metastases and survival. Age, diameter, ulceration, grade, depth of invasion, muscle invasion, mitotic rate, lymphatic-vascular invasion, and tumor infiltrating lymphocytes were evaluated. Two (4.5%) patients had local recurrence, 10 (23%) had metastases to cervical lymph nodes, and 2 (4.5%) died from complications of extensive spread in the neck. One patient with local recurrence had regional node metastases. Regional metastases were discovered at the time of (3 patients) or up to 21 (median 8) months after initial diagnosis, correlating with ulceration (P = 0.003), depth of invasion (P = 0.0001), and mitotic rate (P = 0.05) of the primary tumor. Depth exceeding 2.0 mm. was associated with metastasis (P = 0.028), and all carcinomas deeper than 6.0 mm. metastasized. Eight of the 10 patients with regional metastases had no further disease after treatment through the follow-up period (median 64 months). Both fatalities were older (median 81 years) men. These results underscore the metastatic potential of invasive squamous cell carcinoma of the lip. Histologic analysis of the primary lesion can be used to identify individuals at risk.


Gait analysis and electromyography in fixed- and mobile-bearing total knee replacement: a prospective, comparative study.
The theoretical superiority of mobile-bearing total knee arthroplasties (TKAs) has not yet been proven in clinical studies. The aim of the current study was to compare and to analyse in a patient population that had received either a fixed or a mobile TKA differences in gait analysis electromyography and clinical scores. In a prospective, randomized, patient- and observer-blinded clinical study, 33 patients with primary osteoarthritis of the knee were included. All patients received a Genesis II total knee replacement. Sixteen patients received a mobile and 17 a fixed-bearing cruciate retaining Genesis II TKA. Clinical and quality-of-life scores, electromyography and gait analysis were applied preoperatively and postoperatively with a follow-up of 24 months. In both groups, improvements from pre- to postoperative were detected. whereas the results of gait analysis and electromyography did not show any differences. The results from the clinical and the quality-of-life scores improved from pre- to postoperative, while the Knee Society Score showed a superiority of the mobile-bearing group (mean 159 ± 28) over the fixed-bearing group (mean 134 ± 41). No functional advantage of mobile over fixed-bearing TKA was detected, although the mobile-bearing group had better clinical results for which a reason could not be found. These results only apply to cruciate retaining mobile-bearing TKA with a bearing which allows both rotation and anteroposterior translation.


Proteomic analysis of mdx skeletal muscle: Great reduction of adenylate kinase 1 expression and enzymatic activity.
To investigate the pathophysiological events of Duchenne muscular dystrophy, we analyzed alterations of protein expression in hindlimb muscles of three month old mdx mice using two-dimensional gel electrophoresis and mass spectrometry. About 40 differentially expressed proteins from the cytosolic fraction and 20 from the microsomal fraction of mdx hindlimb muscles were identified. Among these altered proteins, adenylate kinase 1 (AK 1) was particular interesting because its decrease in abundance was so dramatic (> four-fold). Enzymatic assays demonstrated that AK 1 activity was also decreased in mdx mice. Furthermore, the expression and enzymatic activity of AK 1 were consistently reduced in mdx mice at one and six months of age, suggesting a direct relationship between the lack of dystrophin and alteration of AK 1. Along with AK 1, creatine kinase (CK) provides a major pathway for regulation of nucleotide ratios and energy metabolism in muscles. To gain a better understanding of mechanisms of energy metabolism, we also investigated CK activities in these mdx mice at different ages. Our results suggested that decreased AK 1 expression and activity might result in redistribution of energy flow through the alternative CK system, thus a compensatory potential might limit cellular energy failure in mdx skeletal muscle.


Comparative thermodynamics of opioid receptor ligand interaction in the bovine adrenal medulla membranes--evidence of opioid site heterogeneity.
1. A marked dependence on temperature of agonist binding delta, mu and kappa 1-3 opioid sites in the bovine adrenal medulla was observed, at the range of 0 to 37 degrees C. These changes concern kinetic (k1) and equilibrium constants (Kd), but not binding capacities (Bmax). 2. These dependences are different for each ligand and each opioid receptor, suggesting their molecular heterogeneity. 3. The comparative thermodynamics indicates that the interaction of opioid agonists with their receptor is exergonic (delta G degree < 0) and entropy driven (delta S degree > 0). 4. The comparison of Van't Hoff and Arrhenius plots indicates a discrete mechanism in the binding of each opioid receptor.


[How do social-economic differences in residential characteristics affect mortality? A literature review].
Authors review studies on associations between mortality and geographic distribution of social-economic health determinants published in the last 15 years. Hungarian publications after the 1990ies identified several indicators with effect on mortality related to the place of residence such as size of the settlement, population density, joblessness, average income, education, and racial or ethnic mixture. As a rule, these papers analyzed time and geographically aggregated data with simple descriptive statistical approach. Major international studies, however, aside the area indicators commonly included individual data and applied multilevel statistical analysis. Earlier, the incoherent results of some complex statistical analyses were explained by lack of overall health concept, which could utilize effects both at individual and population levels. The most recent multilevel statistical methodology, using information related to spatial correlation and fitted to health models, can help researchers to better understand the causes and effects of health inequities.


Antigenic analyses of cilia-associated respiratory (CAR) bacillus isolates by use of monoclonal antibodies.
Mouse monoclonal antibodies (MAbs) developed to a rat isolate (R-3) of cilia-associated respiratory (CAR) bacillus were used to assess antigenic relationships among three rat and five rabbit CAR bacillus isolates. Evaluation of MAbs by enzyme-linked immunosorbent assays (ELISAs) indicated that 87 of 241 hybridomas secreted CAR bacillus-reactive antibodies that could be grouped into four major groups. Group-I MAbs reacted with epitopes expressed by all CAR bacillus isolates and at least two or more nonrelated species of bacteria. Group-II, -III, and -IV MAbs reacted with only one or more of the rat CAR bacillus isolates; no MAbs reacted only with rat and rabbit CAR bacillus isolates. Western blot analyses indicated that 41-, 50-, and 105-kDa peptides of rat CAR bacillus isolates expressed rat CAR bacillus group- and isolate-specific epitopes. Hyperimmune anti-CAR bacillus antiserum and serum specimens from a CAR bacillus histologically positive mouse and rat also reacted with the 41-, 50-, and 105-kDa peptides. Sera from CAR bacillus histologically negative rats did not react with these peptides. These results suggest that the 41-, 50-, and 105-kDa peptides may represent suitable antigens for development of a specific ELISA for detection of rodent CAR bacillus infections. Furthermore, these data indicate that use of crude CAR bacillus preparations for either rat or rabbit CAR bacillus ELISAs is inappropriate.