BIOS-4 as an embodiment of CELSS development conception.
Any attempt to create LSS for practical applications must take into account the possibility of castastrophic consequences if the problem of LSS reliability and stability is not solved. An integrated conception of CELSS studies development as a possible way to increase its reliability is considered. The BIOS-4 facility project is developed in the context of the conception. Three principles of highly effective experimental CELSS facility design are proposed. Some details of BIOS-4 design and its exploitation features are presented.


Hospital-acquired viral pathogens in the neonatal intensive care unit.
Hospital-acquired infections caused by viruses are a cause of considerable morbidity and occasional mortality in critically ill neonates. The intensive care environment allows for efficient spread of viral pathogens, and secondary cases among both patients and healthcare workers are frequently observed. We review the common viral causes of hospital-acquired infections in neonates, including rotavirus, respiratory syncytial virus, and others, discuss epidemiology and clinical syndromes, and summarize recommendations for control in outbreak situations. Chemoprophylaxis, isolation procedures, and care of affected staff are also addressed.


Density cumulant functional theory from a unitary transformation: N-representability, three-particle correlation effects, and application to O4(+).
A new approach to density cumulant functional theory is developed that derives density cumulant N-representability conditions from an approximate Fock space unitary transformation. We present explicit equations for the third- and fourth-order two-particle cumulant N-representability, as well as the second-order contributions that depend on the connected three-particle density cumulant. These conditions are used to formulate the ODC-13 method and the non-iterative (λ3) correction that employ an incomplete description of the fourth-order two-particle cumulant N-representability and the second-order three-particle correlation effects, respectively. We perform an analysis of the ODC-13 N-representability description for the dissociation of H2 and apply the ODC-13 method and the (λ3) correction to diatomic molecules with multiple bond character and the symmetry-breaking tetraoxygen cation (O4(+)). For the O4(+) molecule, the vibrational frequencies of the ODC-13(λ3) method do not exhibit spatial symmetry breaking and are in a good agreement with the recent infrared photodissociation experiment. We report the O4(+) equilibrium structure, harmonic frequencies, and dissociation energy computed using ODC-13(λ3) with a diffuse, core-correlated aug-cc-pCVTZ basis set.


Is obstructive sleep apnea a risk factor for diabetes?
Obstructive sleep apnea (OSA) and type 2 diabetes are both closely related to obesity and their prevalence is increasing due to the rising average body weight in Western countries. The findings of epidemiological studies have implicated that OSA increases the risk for cardiovascular disease, and metabolic disturbances, such as insulin resistance, may link OSA to vascular morbidity. A number of observational clinical studies have evaluated the relationship between OSA and insulin resistance, suggesting an independent association. However, the confounding effect of obesity complicates the establishment of a causal relationship between OSA and insulin resistance. Potential mechanisms that may underpin this relationship were evaluated in animal and human experimental studies and include intermittent hypoxia, arousals from sleep with concomitant sympathetic activation and sleep fragmentation. Currently only three randomized controlled trials investigating the effects of OSA on insulin resistance have been published. In these trials OSA patients were randomly assigned to treatment with continuous positive airway pressure (CPAP) or subtherapeutic CPAP and treatment effects on various measures of insulin resistance were examined. In two of these trials there was no effect of CPAP on glucose metabolism and in one trial a small beneficial effect of CPAP was observed. Further carefully conducted clinical studies and randomized controlled interventional CPAP trials are needed to determine the extent to which OSA is a risk factor for diabetes and its effect on glucose metabolism.


DNA damage in HeLa s3 cells by an antitumor drug Ledakrin and other antitumor 1-nitro-9-aminoacridines.
Ledakrin and seven other antitumor and cytotoxic derivatives of 1-nitro-9-aminoacridine were shown to induce DNA-single strand breaks in HeLa S3 cells as found by alkaline sucrose gradient centrifugation. The induced DNA damage is of non-random character. Some of Ledakrin -induced DNA breaks are probably generated by endonucleolytic cleavage in the course of repair processes as indicated by experiments with Novobiocin, an antibiotic preventing the incision step of DNA repair. Other Ledakrin -induced DNA breaks observed on alkaline sucrose gradients may arise from alkali-labile sites in DNA. Most of such sites seem to be converted to breaks after brief exposure to alkali. The extent of DNA damage by 1-nitro-9-aminoacridines was found to be correlated with cytotoxic activities of these compounds against HeLa S3 cells. Furthermore, Ledakrin and other derivatives seem to induce DNA-repair synthesis in HeLa S3 cells as judged by the stimulation of hydroxyurea (HU)-resistant incorporation of [3H] thymidine into DNA. The agents studied differ in their concentrations required to produce a considerable stimulation of DNA repair, whereas the maximal level of this effect is similar for all the derivatives assayed. The former values are correlated with cytotoxic activities of these compounds and seem to reflect the overall extent of DNA damage by 1-nitro-9-aminoacridines. Stimulation of DNA-repair synthesis is gradually shut off during prolonged incubation of the cells with Ledakrin or during postincubation of the cells in a drug-free medium. Such postincubation results also in the gradual accumulation of DNA-single strand breaks as observed by alkaline sucrose centrifugation. Hence, HeLa S3 cells are incapable of efficiently removing DNA damage by 1-nitro-9-aminoacridines, though the drug's action activates temporarily some repair mechanisms. The reported results suggest that overall DNA damage may contribute to the cytotoxic effects of 1-nitro-9-aminoacridines besides previously found ability of these agents to form interstrand DNA cross-links.


Lateralization patterns of verbal stimuli processing assessed by reaction time and event-related potentials in schizophrenic patients.
Event-related potentials (ERPs), reaction time (RT), number of errors and number of omissions were recorded by using a visual target detection paradigm in which consonant pairs were presented in a central and in a lateral condition. Fourteen DSM III drug-free schizophrenics and 19 healthy subjects took part in the study. All of them were male and right-handed. Independent principal component analysis and varimax rotation were performed for each condition. Separate ANOVAs were performed on factor scores computed for each rotated principal component. In the central condition, schizophrenic patients showed a reduction of the late positive complex (LPC) peak with respect to normal controls, and this reduction was associated with a predominant negative symptomatology. No pattern of lateral asymmetry was found in either group for behavioural and ERP measures. For the lateral condition, a significant advantage of the right visual field (RVF) presentation (shorter RT and larger LPC peak for RVF stimuli) was observed in normal subjects, while no visual field effect on these two measures was detected in schizophrenics. Significant differences were found between the two groups on both behavioural and ERP measures for the RVF only. Furthermore, two ERP components showed a marked advantage of the left visual field in the patient group, associated with a predominance of positive symptoms in the clinical picture.


Biocatalytic, one-pot diterminal oxidation and esterification of n-alkanes for production of α,ω-diol and α,ω-dicarboxylic acid esters.
Direct and selective terminal oxidation of medium-chain n-alkanes is a major challenge in chemistry. Efforts to achieve this have so far resulted in low specificity and overoxidized products. Biocatalytic oxidation of medium-chain n-alkanes - with for example the alkane monooxygenase AlkB from P. putida GPo1- on the other hand is highly selective. However, it also results in overoxidation. Moreover, diterminal oxidation of medium-chain n-alkanes is inefficient. Hence, α,ω-bifunctional monomers are mostly produced from olefins using energy intensive, multi-step processes. By combining biocatalytic oxidation with esterification we drastically increased diterminal oxidation upto 92mol% and reduced overoxidation to 3% for n-hexane. This methodology allowed us to convert medium-chain n-alkanes into α,ω-diacetoxyalkanes and esterified α,ω-dicarboxylic acids. We achieved this in a one-pot reaction with resting-cell suspensions of genetically engineered Escherichia coli. The combination of terminal oxidation and esterification constitutes a versatile toolbox to produce α,ω-bifunctional monomers from n-alkanes.


[The therapy for acute acquired syphilitic uveitis].
In retrospective study (analysis) to report the effectiveness of therapy on anatomic and functional outcomes in patients with acute acquired syphilitic uveitis. Retrospective case-control study. 3 patients (4 eyes) with symptoms of ocular acute inflammation which occurred in latent stage of syphilis are included in our study. In all patients uveitis was the first clinical symptom of syphilis. In all 3 patients the infection of acquired syphilis was diagnosed using specific and non-specific tests for treponemal antibody. In 2 eyes we performed diagnostic and therapeutic pars plana vitrectomy. The management of ocular syphilis in all patients consisted of mega units of aqueous penicillin intravenously or intramusculary, in 2 patients we used the combination with steroids. In 2 eyes with perioperatively obtained material of vitreous (during pars plana vitrectomy) specific treponemal antibody was detected using specific (TPHA) test. In the third patient infection was verified serologically. The final visual acuity of all 4 eyes is better than or equal to 6/12. The visual acuity of one exe is 6/6. Uveitis and neuroretinitis may be the first symptom of acquired syphilis in latent stage. Early antibiotic treatment significantly improves anatomical reparation of infection and visual outcomes.


Preparation of 3 beta, 5 alpha-, 3 alpha, 5 alpha- and 3 alpha, 5 beta-tetrahydro derivatives of 19-noraldosterone by chemical synthesis and microbial bioconversion.
The 3 beta, 5 alpha-, 3 alpha, 5 alpha- and 3 alpha, 5 beta-tetrahydro derivatives 19, 20 and 27 of 19-noraldosterone (1) were prepared to facilitate the search for these compounds in urine. The diketal 4, consisting of a 2:1 mixture of the 5,6- and 5(10)-ene isomers, was hydrogenated with Pd-C and partially hydrolyzed to 5 alpha, 10 alpha- and 5 alpha, 10 beta-dihydroketals 8 and 10 in a 1:2.5 ratio. Assignment of protons was done with aid of COSY 45 experiments. Compound 10 was reduced with diisobutylaluminum hydride (DIBAH) to 4 products: the 3 alpha- and 3 beta-ol hemiacetals 16 and 15, and the corresponding tetraols 14 and 13. Alternatively, hydrogenation of the 4-en-3-one 2 gave 10, its 5 beta, 10 beta-isomer 21 and the tetrahydro compound 22, in a 4:2:1 ratio. A better way to prepare the 5 beta, 10 beta-series involved microbial conversion of 2 with Clostridium paraputrificum, and the resulting tetrahydrolactone 23 was reduced with DIBAH to the hemiacetal 24. Acid hydrolysis of 16, 15 and 24 afforded 20, 19 and 27, respectively. According to [1H]-NMR, in solution 20 and 24 exist as mixtures of isomers, while 19 appears in one form only. Periodate oxidation converted 19 and 27 into their gamma-etiolactones 18 and 28. EI MS base peaks are different and characteristic for 19, 20 and 27.


Vestibular hypofunction in the initial postoperative period after surgical treatment of superior semicircular canal dehiscence.
1) Determine the prevalence of vestibular hypofunction in the immediate postoperative period after surgical treatment of superior semicircular canal dehiscence syndrome. 2) Evaluate whether dehiscence length is associated with risk of postoperative vestibular hypofunction. 3) Compare the prevalences of immediate and late postoperative vestibular hypofunction. Clinical review. Tertiary referral center. Subjects with superior canal dehiscence syndrome (n = 42) based on history, physiologic testing, and computed tomography findings, who underwent middle fossa craniotomy and superior canal dehiscence plugging. Dehiscence length was measured intraoperatively. Bedside horizontal head thrust testing (hHTT) was administered between postoperative days 1 to 7 to diagnose immediate postoperative vestibular hypofunction. Both hHTT and quantitative vestibulo-ocular reflex testing were administered 6 to 29 weeks postoperatively to detect late vestibular hypofunction. Dehiscence length and hypofunction in response to hHTT. Thirty-eight percent of the subjects (95% confidence interval, 25-54) had hypofunction in response to hHTT within 1 week after surgery. Mean dehiscence lengths were 4.9 (range, 2.0-10.5 mm) and 3.4 mm (range, 1.0-5.5 mm) in subjects with and without postoperative hypofunction, respectively (p = 0.0018). Each 1-mm increase in dehiscence length increased the odds of immediate postoperative hypofunction 2.6-fold (95% confidence interval, 1.3-5.1). The prevalence of vestibular hypofunction was significantly higher in the early compared with the late postoperative period. Immediate postoperative vestibular hypofunction is common, particularly with larger dehiscences. This hypofunction may typically resolve, given that the prevalence of vestibular hypofunction 6 weeks postoperatively is low. Possible mechanisms include intraoperative loss of perilymph, which may be more likely with larger dehiscences.


Selected suppliers of reagents and equipment.
This appendix lists the full contact information, including website URLs, for all suppliers mentioned in Current Protocols in Molecular Biology.


Stability of absolute copy number of housekeeping genes in preeclamptic and normal placentas, as measured by digital PCR.
Measuring mRNA expression is fundamental to placental research. Ideally, mRNA transcript numbers are directly quantified. However, PCR analysis using the ΔΔCT method relies on the stability of housekeeping genes and only reports relative expression. Digital PCR (dPCR) directly quantifies mRNA copy number and is more accurate than quantitative PCR. We quantified absolute mRNA copy number of housekeeping genes in normotensive pre-term (n = 20), severe preeclamptic (n = 11) and term (n = 12) placenta using dPCR. Whilst there was some variation, we confirm absolute mRNA copy number of GAPDH, TOP1, CYC1 and YWHAZ in placenta does not significantly alter between these cohorts, or across gestation.


Subclinical left ventricular dysfunction in asymptomatic patients with Type II diabetes mellitus, related to serum lipids and glycated haemoglobin.
The aim of the present study was to measure regional ventricular function at rest and during stress in order to assess if patients with Type II diabetes have subclinical myocardial dysfunction and if it is related to risk factors. Seventy subjects (35 patients with Type II diabetes with no symptoms, signs or history of heart disease, and 35 age- and sex-matched healthy controls) had echocardiography at rest and during dobutamine stress. Myocardial velocities were measured off-line from digital loops of colour tissue Doppler. Subendocardial function was assessed from the mean longitudinal velocities of four basal segments (apical views) and radial function from the velocities of the basal posterior wall (parasternal view). Systolic functional reserve was calculated as the increase in velocity from baseline. Longitudinal peak systolic velocity was lower in patients with diabetes, at rest (5.6 +/- 1.4 compared with 6.5 +/- 1.1 cm/s) and at peak stress (10.9 +/- 2.8 compared with 14.3 +/- 2.1 cm/s) (both P <0.01). Functional reserve was impaired in patients with diabetes (+5.4 +/- 2.0 compared with +7.7 +/- 1.7 cm/s; P <0.01). Radial systolic velocity was higher in patients with diabetes (5.4 +/- 1.3 compared with 4.7 +/- 1.4 cm/s; P <0.05). Resting longitudinal systolic function correlated inversely with low-density lipoprotein-cholesterol ( r =-0.53), glycated haemoglobin ( r =-0.48), age ( r =-0.41) and diastolic blood pressure ( r =-0.38) (all P < 0.05). Peak stress systolic velocity correlated inversely with glycated haemoglobin ( r =-0.46) and age ( r =-0.44) (both P < 0.01). In conclusion, patients with Type II diabetes and no clinical heart disease have impaired subendocardial function of the left ventricle at rest and peak stress, which is related to glycated haemoglobin and serum low-density lipoprotein-cholesterol.


Novel perspectives in cancer therapy: Targeting ion channels.
By controlling ion fluxes at multiple time scales, ion channels shape rapid cell signals, such as action potential and synaptic transmission, as well as much slower processes, such as mitosis and cell migration. As is currently increasingly recognized, a variety of channel types are involved in cancer hallmarks, and regulate specific stages of neoplastic progression. Long-term in vitro work has established that inhibition of these ion channels impairs the growth of cancer cells. Recently, these studies have been followed up in vivo, hence revealing that ion channels constitute promising pharmacological targets in oncology. The channel proteins can be often accessed from the extracellular milieu, which allows use of lower drug doses and decrease untoward toxicity. However, because of the central physiological roles exerted by ion channels in excitable cells, other types of side effects may arise, the gravest of which is cardiac arrhythmia. A paradigmatic case is offered by Kv11.1 (hERG1) channels. HERG1 blockers attenuate the progression of both hematologic malignancies and solid tumors, but may also lead to the lengthening of the electrocardiographic QT interval, thus predisposing the patient to ventricular arrhythmias. These side effects can be avoided by specifically inhibiting the channel isoforms which are highly expressed in certain tumors, such as Kv11.1B and the neonatal forms of voltage-gated Na(+) channels. Preclinical studies are also being explored in breast and prostate cancer (targeting voltage-gated Na(+) channels), and gliomas (targeting CLC-3). Overall, the possible approaches to improve the efficacy and safety of ion channel targeting in oncology include: (1) the development of specific inhibitors for the channel subtypes expressed in specific tumors; (2) drug delivery into the tumor by using antibodies or nanotechnology-based approaches; (3) combination regimen therapy and (4) blocking specific conformational states of the ion channel. We believe that expanding this relatively neglected field of oncology research might lead to unforeseen therapeutic benefits for cancer patients.


A naturalistic, randomized, controlled trial combining cognitive remediation with cognitive-behavioural therapy after first-episode non-affective psychosis.
Cognitive remediation (CR) preceding cognitive-behavioural therapy for psychosis (CBTp) was trialled within routine clinical services, with the hypothesis that following first-episode non-affective psychosis CR would enhance CBTp efficacy by improving neuropsychological performance. A total of 61 patients with DSM-IV non-affective psychoses waiting for routine CBTp were randomized to computerized CR over 12 weeks, supported by a trained support worker, or time-matched social contact (SC). Primary outcome was the blind-rated Psychotic Symptoms Rating Scale (PSYRATS). Secondary outcomes included measures of CBTp progress, cognition, symptoms, insight and self-esteem: all at baseline, after CR (12 weeks) and after CBTp (42 weeks). PSYRATS and global neuropsychological efficacy were tested using mixed-effects models with a group × time interaction term. Measures of CBTp progress and some neuropsychological measures were modelled by regression. There was no significant difference between the CR and SC groups in PSYRATS (group × time coefficient 0.3, 95% confidence interval -0.4 to 1.1, p = 0.39). However, after CR CBTp was shorter [median 7 sessions, interquartile range (IQR) 2-12 after CR; median 13, IQR 4-18 after SC; model p = 0.011] and linked to better insight (p = 0.02). Global cognition did not improve significantly more after CR (p = 0.20) but executive function did (Wisconsin Card Sort, p = 0.012). CBTp courses preceded by CR were far shorter but achieved the same outcome as CBTp preceded by an active control, consistent with neuropsychological improvement enhancing CBTp. CR was delivered by staff with minimal training, offering the potential to reduce the costs of CBTp considerably.


Modelling the signal delivered by a population of first-order neurons in a moth olfactory system.
A statistical model of the population of first-order olfactory receptor neurons (ORNs) is proposed and analysed. It describes the relationship between stimulus intensity (odour concentration) and coding variables such as rate and latency of the population of several thousand sex-pheromone sensitive ORNs in male moths. Although these neurons likely express the same olfactory receptor, they exhibit, at any concentration, a relatively large heterogeneity of responses in both peak firing frequency and latency of the first action potential fired after stimulus onset. The stochastic model is defined by a multivariate distribution of six model parameters that describe the dependence of the peak firing rate and the latency on the stimulus dose. These six parameters and their mutual linear correlations were estimated from experiments in single ORNs and included in the multidimensional model distribution. The model is utilized to reconstruct the peak firing rate and latency of the message sent to the brain by the whole ORN population at different stimulus intensities and to establish their main qualitative and quantitative properties. Finally, these properties are shown to be in agreement with those found previously in a vertebrate ORN population. This article is part of a Special Issue entitled: Neural Coding.


Endocrine carcinoma of the stomach: clinicopathological analysis of 27 surgically treated cases in a single institute.
Gastric endocrine carcinoma (EC) is an uncommon tumor of the stomach and the clinical features are not well known. Additionally, the classification and staging systems of this tumor are not yet unified worldwide. In this study, we reviewed 27 patients with gastric EC to evaluate the clinicopathological characteristics of this tumor. We retrospectively reviewed 27 patients with gastric EC among 6466 patients who had undergone gastrectomy between 1986 and 2008 at our institute. Clinicopathological features including immunohistochemistry of Ki-67 were investigated to evaluate the malignant potential of the tumor. Furthermore, survivals were compared between the 7th edition of the International Union Against Cancer (UICC)-TNM (7th TNM) classification for gastric cancer (GC) and the new TNM classification for foregut neuroendocrine tumors (NET). The median survival of the patients was 19.0 months. The 5-year survival rate was 100% in pathological stage (pStage) I, 40% in pStage II, 38% in pStage III, and 11% in pStage IV according to the 7th TNM classification for GC. Survivals by stage showed great difference between the 7th TNM classification for GC and the new TNM classification for foregut NET, but each system correlated with survival. The Ki-67 labeling index was more than 20% in most of the patients. Univariate analysis revealed that maximum tumor diameter, tumor depth, lymph node metastasis, lymphatic invasion, pStage, and curability had significant correlations with survival. Early detection and curative operations are essential for improving the prognosis of gastric EC. However, some adjuvant chemotherapies are required for advanced-stage tumors. Classification and staging systems may need to be unified worldwide for further analysis.


Comparison of pathophysiologic changes in the lungs of calves challenge exposed with Escherichia coli-derived endotoxin and Pasteurella haemolytica, alone or in combination.
Pulmonary responses to intratracheal challenge exposure with Pasteurella haemolytica, with or without Escherichia coli-derived endotoxin, E coli endotoxin alone, or saline solution were compared in anesthetized, mechanically ventilated neonatal calves. Baseline values for dynamic compliance, total pulmonary resistance, functional residual capacity, arterial blood gas tensions, hemogram, leukogram, and systemic and pulmonary arterial pressures were recorded for each calf. After baseline data were obtained, calves were challenge exposed with logarithmic-growth phase P haemolytica organisms with or without E coli endotoxin, E coli endotoxin alone, or saline solution (0.9% NaCl). Physiologic data were obtained immediately after challenge exposure and at various intervals over the next 6 hours. Calves challenge exposed with P haemolytica alone developed sever hypoxemia, had increased alveolar-arterial oxygen difference and threefold increases in total pulmonary resistance, became hypercarbic, had decreased functional residual capacity, and developed systemic hypotension without change in pulmonary arterial pressure. At necropsy, these calves had extensive multifocal areas of necrohemorrhagic and purulent pneumonia. Ratio of extravascular lung water to lung dry weight was not significantly increased in lung specimens obtained from calves challenge exposed with P haemolytica, but ratio of lung wet weight to dry weight was increased, indicating that increased lung wet weight was attributable largely to increased solids and not to fluid alone. (Extravascular lung water measurement excludes fluid from the vascular compartment.) Intratracheal challenge exposure with endotoxin failed to alter lung function and caused minor changes in lung structure consisting of focal areas of hemorrhage and edema.(ABSTRACT TRUNCATED AT 250 WORDS)


Diffusion-weighted magnetic resonance imaging is effective to detect ileocolonic ulcerations in Crohn's disease.
Magnetic resonance entero-colonography enables accurate assessment of ileocolonic Crohn's disease, but the need for bowel cleansing and rectal enema limits considerably its use in daily practice. We evaluated the accuracy of diffusion-weighted magnetic resonance entero-colonography with neither bowel cleansing nor rectal enema to assess endoscopic activity. Forty-four Crohn's disease patients underwent prospectively and consecutively diffusion-weighted magnetic resonance entero-colonography [with apparent diffusion coefficient (ADC) and Clermont score calculation] and ileocolonoscopy [with Crohn's Disease Endoscopic Index of Severity (CDEIS) and Simplified Endoscopic score for Crohn's Disease (SES-CD) calculation]. Mean ADC was inversely correlated with total CDEIS (ρ = -0.40; P = 0.0067) and total SES-CD (ρ = -0.33; P = 0.032). Considering the 194 segments, ADC was inversely correlated with segmental CDEIS (-0.48; P < 0.001) and segmental SES-CD (-0.44; P < 0.001). ADC values were lower in segments with deep ulcers (1.30 ± 0.23) or superficial ulcerations (1.75 ± 0.64) than in non-ulcerated segments (2.15 ± 0.5) (P = 0.001). Using a receiver operating curve, we determined that segmental ADC <1.42 detected endoscopic deep ulcerations with sensitivity = 0.91 and specificity = 0.83 (Area under the curve = 0.84; P < 0.001). Segmental ADC <1.88 detected endoscopic superficial ulcerations with sensitivity = 0.64 and specificity = 0.75. The segmental ADC values decreased when the ulcerations size increased (P = 0.0001). Clermont score correlated with ileal CDEIS (0.63; P < 0.05) and ileal SES-CD (0.58; P < 0.05). Clermont score was higher in ulcerated segments (23.3 ± 8.4) than in non-ulcerated segments (12.4 ± 10.0) (P = 0.006) and increased with ulcers size (P = 0.012). Clermont score >18.9 detected ulcerations with sensitivity = 0.79 and specificity = 0.73. Diffusion-weighted magnetic resonance entero-colonography using apparent diffusion coefficient and Clermont score was effective to indirectly detect endoscopic ulcerations in ileocolonic Crohn's disease.


Edrophonium antagonizes combined lidocaine-pancuronium and verapamil-pancuronium neuromuscular blockade in cats.
The effects of lidocaine or verapamil on pancuronium neuromuscular blockade and the ability of anticholinesterase agents to antagonize these combined blockades were studied in 14 cats using a standard peroneal nerve-anterior tibialis muscle preparation. Pancuronium was infused at a constant rate to produce a stable 50% depression of single twitch tension. In nine cats, intravenous lidocaine boluses followed by a constant infusion produced serum lidocaine levels of 5.09 +/- 1.9 micrograms/ml (mean +/- SD) and resulted in an additional 20.0 +/- 5.5% depression of twitch tension. In the other five cats, intravenous injection of 0.15 mg/kg of verapamil produced an additional 12.8 +/- 8.0% twitch depression of pancuronium-induced neuromuscular blockade. For individual animals, edrophonium antagonism of the combined lidocaine-pancuronium-induced neuromuscular blockade or combined verapamil-pancuronium-induced neuromuscular blockade was not significantly different from antagonism of an equivalent twitch depression produced by pancuronium alone. It is concluded that lidocaine and verapamil augment neuromuscular blockade caused by pancuronium and that anticholinesterase antagonism of this augmented blockade can be expected to occur in a normal fashion.


Interactions of protoporphyrin IX and its derivatives with benzodiazepine receptor.
The aim of this study was to determine binding affinities of the Peripheral benzodiazepine receptor (PBR) with protoporphyrin IX, haematoporphyrin (Hp), and two arginized derivatives: Hp(Arg)(2) and PP(Arg)(2). The quenching of protein fluorescence was used to measure association constants during titration of PBR solution with PP derivatives in a function of concentration. The experimental data was fitted to theoretical curves, assuming strong and weak binding model with one binding site. As a result of this early study affinity constants were determined. The highest affinity toward PBR showed PPIX and Hp, Hp(Arg)(2), PP(Arg)(2) lower, respectively. Changes in pH of protein solution resulted in decrease of association constants in nearly all examined derivatives.


Initial perspectives on process threat management.
Terrorist and criminal acts are now considered credible risks in the process industries. Deliberate attacks on the nation's petroleum refineries and chemical plants would pose a significant threat to public welfare, national security, and the US economy. To-date, the primary response of government and industry has been on improved security to prevent attacks and the associated consequences. While prevention is clearly preferred, the potential for successful attacks must be addressed. If plant security is breached, the extent of the inflicted damage is determined by the available plant safety systems and procedures. We refer to this "inside the gate" response as process threat management. The authors have initiated a joint industry/academia study to address: the level of safety provided by existing plant equipment and safety systems in response to a terrorist act, and identification of process (rather than security) needs or opportunities to address this new safety concern. This paper describes the initial perspectives and issues identified by the team at the beginning of the study.


An Active Drimane-Type Lactone from Polygonum jucundum Attenuates Lipopolysaccharide-Induced Acute Lung Injury in Mice Through TLR4-MAPKs Signaling Pathway.
The herbs of Polygonum jucundum Lindex. (Polygonaceae) is a traditional Chinese medicine for inflammatory diseases. 2α-Hydroxyl-3β-angeloylcinnamolide (HAC), a drimane-type sesquiterpenoid, was the major active compound of the ethanol extract of P. jucundum which inhibited the production of inflammatory mediators. However, the biological mechanism of HAC for anti-inflammatory activity has not been reported. In the current study, we investigated whether HAC could suppress the production of inflammatory mediators in lipopolysaccharide (LPS)-induced acute lung injury in mice (ALI) through downregulation of Toll-like receptor 4 (TLR4) and activations of mitogen-activated protein kinases (MAPKs) and inducible protein nitric oxide synthase (iNOS). Moreover, our data indicated that HAC inhibits the overexpression of iNOS and TLR4 in LPS-treated RAW264.7, and also inhibits MAPK signal. These findings suggest that HAC shows anti-inflammatory effects in ALI mice through suppressing TLR4-mediated MAPK pathway in activated macrophages. In addition, six derivatives of HAC obtained by structure modification were investigated for their inhibitory effects on the production of nitric oxide (NO) and tumor necrosis factor-α (TNF-α), suggesting that the acetylation could increase the inhibition of HAC on TNF-α release in LPS-treated RAW264.7 cells. In summary, all these results showed that HAC may be a potential anti-inflammatory lead compound for the treatment of acute lung injury.


Effects of increasing diet fermentability on intake, digestion, rumen fermentation, blood metabolites and milk production of heat-stressed dairy cows.
Heat stress is a major problem for dairy cows in hot climates, thus coping strategies are essential. This study evaluated the effects of increasing diet fermentability on intake, total tract digestibility, ruminal pH and volatile fatty acids (VFA) profile, blood metabolite profile and milk production and composition of lactating dairy cows managed under conditions of ambient heat stress. Nine multiparous cows (650 ± 56 kg BW; mean ± SD) averaging 102 ± 13 days in milk and producing 54 ± 6 kg/day were randomly assigned to a triplicate 3 × 3 Latin square. During each 21-day period, cows were offered one of three total mixed rations that varied in diet fermentability. The three levels of diet fermentability were achieved by increasing the proportion of pellets containing ground wheat and barley in the dietary DM from 11.7% (low), to 23.3% (moderate), and 35.0% (high) by replacing ground corn grain. Each period had 14 day of adaptation and 7 day of sampling. The ambient temperature-humidity index ( ≥ 72) indicated that the cows were in heat stress almost the entire duration of the study. Also, rectal temperature of cows was elevated at 39.2°C, another indication of heat stress. Increasing diet fermentability linearly decreased dry matter intake (22.8, 22.5, 21.8 kg/day for low, moderate and high, respectively; P ≤ 0.05) but increased non-fibre carbohydrate digestibility (P ≤ 0.05) and tended to increase digestibility of DM (P = 0.10) and crude protein (P = 0.06). As a result, the intake of digestible DM was not affected by the treatments. The production of 3.5% fat corrected milk (32.6, 33.7, and 31.5 kg/day) was quadratically (P ≤ 0.05) affected by diet fermentability with lower production for the high diet compared with the other two, which were similar. Rumen pH (ruminocentesis) and proportions of butyrate and isovalerate linearly decreased whereas propionate proportion linearly increased with increasing diet fermentability (P ≤ 0.05). The rumen concentration of NH3-N (11.0, 9.0, and 8.7 mg/dL) and blood concentration of urea linearly decreased with increasing diet fermentability (P ≤ 0.05). The activity of alkaline phosphatase increased (65.1, 83.2, and 84.9 U/l) and concentration of malondialdehyde decreased (2.39, 1.90 and 1.87 µmol/l) linearly with increasing diet fermentability (P ≤ 0.05), which indicated possible attenuation of the effects of oxidative stress with increasing diet fermentability. Overall, a modest increase of diet fermentability improved nitrogen metabolism, milk protein production and oxidative stress of heat-stressed dairy cows, but a further increase in diet fermentability decreased milk yield.


[Osmotic behavior of mouse embryonic cells subjected to hypotonic shock].
Osmotic adaptation in a blastomere of mouse embryo has been studied by the direct measurement of the cell volume using laser scanning microscopy microtomography followed by quantitative 3D reconstruction. Embryonic cells subjected to hypotonic shock first swelled and then returned to the initial size. At the beginning of osmotic stress, the swelling occurred by the van't Hoff equation with the water permeability coefficient (L(p)) of 0.4 micro x min(-1) atm(-1). The phase of the regulatory volume decrease was not abolished by Na+/K(+)-ATPase inhibition.


Chondroitin sulphate proteoglycans in the CNS injury response.
As the preceding discussion has demonstrated, experimental data now indicate that the expression of a number of different CSPGs is increased following CNS injury. The hyalectans neurocan, versican and [figure: see text] brevican, plus NG2 and phosphacan are upregulated following injury and all have been shown to exhibit inhibitory effects on neurite outgrowth in vitro. It is likely therefore that the increased expression of these molecules contributes to the non-permissive nature of the glial scar. The relative contributions of individual molecules remain, however, to be determined. It is important to remember also that not only does the glial scar contain many different inhibitory molecules, but that these are the products of a number of different cells, including not just astrocytes, but also oligodendrocyte progenitor and meningeal cells. It is arguable that the latter two cell types make a greater contribution than astrocytes to the inhibitory environment of the injured CNS. Recently, attempts have been made to alter the CSPG component of the glial scar in the hope that this will facilitate improved axonal regeneration. Three studies (Bradbury et al., 2002; Yick et al., 2000; Moon et al., 2001) have reported an improved regenerative response following treatment of the injured CNS with chondroitinase ABC. CSPGs represent a significant source of inhibition within the injured CNS; these studies indicate that successful CNS regeneration may be brought about by interventions which target these molecules and/or the cells which produce them.


Effect of lung volume reduction surgery on gas exchange and pulmonary hemodynamics at rest and during exercise.
Lung volume reduction surgery (LVRS) has become an extended surgery for emphysema in order to improve the dyspnea of severely affected patients. Because resection of lung areas may reduce the vascular bed, which is an important factor of pulmonary hypertension in emphysematous patients, especially during exercise, the aim of our study was to assess the outcome of pulmonary hemodynamics and gas exchange at rest and during exercise after LVRS. Nine patients had right heart catheterization before and 3 to 12 mo (mean, 4.5 mo) after LVRS. FEV1 increased from 705 to 1,005 ml (p < 0.05) after LVRS. PaO2, PaCO2 and mean pulmonary artery pressure (Ppa) did not change after LVRS, either at rest or during exercise. However, a significant overall decrease of the respiratory swings of the pulmonary artery diastolic pressure (DeltaPd) at rest (median value, from 12 to 8 mm Hg, p < 0.01) and during exercise (from 20 to 15 mm Hg, p < 0.05) was observed. There was a significant correlation between the change in resting Ppa (Ppa before minus Ppa after LVRS) and the change in resting DeltaPd (r = 0.73, p < 0.03), and also between the change in exercising Ppa and the change in resting DeltaPd (r = 0.80, p < 0.02). Significant correlations were also found between the change in exercising Ppa and the change in exercising PaO2 (r = -0.70, p < 0.05), and between the change in exercising Ppa and the change in exercising PaCO2 (r = 0.76, p < 0. 03). We conclude that pulmonary hemodynamics in most cases are not impaired by LVRS either at rest or during exercise. The possible mechanisms influencing hemodynamics after a lung volume reduction procedure are discussed.


Microenvironmental factors involved in human amnion mesenchymal stem cells fate decisions.
Human amnion mesenchymal stem cells (HAMCs) show great differentiation and proliferation potential and also other remarkable features that could serve as an outstanding alternative source of stem cells in regenerative medicine. Recent reports have demonstrated various kinds of effective artificial niche that mimic the microenvironment of different types of stem cell to maintain and control their fate and function. The components of the stem cell microenvironment consist mainly of soluble and insoluble factors responsible for regulating stem cell differentiation and self-renewal. Extensive studies have been made on regulating HAMCs differentiation into specific phenotypes; however, the understanding of relevant factors in directing stem cell fate decisions in HAMCs remain underexplored. In this review, we have therefore identified soluble and insoluble factors, including mechanical stimuli and cues from the other supporting cells that are involved in directing HAMCs fate decisions. In order to strengthen the significance of understanding on the relevant factors involved in stem cell fate decisions, recent technologies developed to specifically mimic the microenvironments of specific cell lineages are also reviewed. Copyright © 2015 John Wiley & Sons, Ltd.


Changes in methylation within the STK32B promoter are associated with an increased risk for generalized anxiety disorder in adolescents.
Generalized anxiety disorder (GAD) is highly prevalent among adolescents. An early detection of individuals at risk may prevent later psychiatric condition. Genome-wide studies investigating single nucleotide polymorphisms (SNPs) concluded that a focus on epigenetic mechanisms, which mediate the impact of environmental factors, could more efficiently help the understanding of GAD pathogenesis. We investigated the relationship between epigenetic shifts in blood and the risk to develop GAD, evaluated by the Development and Well-Being Assessment (DAWBA) score, in 221 otherwise healthy adolescents. Our analysis focused specifically on methylation sites showing high inter-individual variation but low tissue-specific variation, in order to infer a potential correlation between results obtained in blood and brain. Two statistical methods were applied, 1) a linear model with limma and 2) a likelihood test followed by Bonferroni correction. Methylation findings were validated in a cohort of 160 adults applying logistic models against the outcome variable "anxiety treatment obtained in the past" and studied in a third cohort with regards to associated expression changes measured in monocytes. One CpG site showed 1% increased methylation in adolescents at high risk of GAD (cg16333992, padj. = 0.028, estimate = 3.22), as confirmed in the second cohort (p = 0.031, estimate = 1.32). The identified and validated CpG site is located within the STK32B promoter region and its methylation level was positively associated with gene expression. Gene ontology analysis revealed that STK32B is involved in stress response and defense response. Our results provide evidence that shifts in DNA methylation are associated with a modulated risk profile for GAD in adolescence.


Congenital hypofibrinogenemia and pregnancy, obstetric and hematological management.
A 27-year-old pregnant woman with severe congenital hypofibrinogenemia was studied from the 18th to the 40th week of pregnancy, after which she had a normal delivery. The results of the quantitative and qualitative fibrinogen studies done on the patient made it possible to rule out associated dysfibrinogenemia. No variations in the concentration and function of fibrinogen were observed during gestation. The only treatment given was a transfusion of fresh plasma prior to delivery. The importance of fibrinogen in maintaining normal placental insertion and the obstetric management of predelivery and delivery are discussed.


Mid-term follow-up of 183 arterial myocardial revascularization procedures.
To evaluate the mid-term results of complete arterial myocardial revascularization performed with arterial conduits. From July 1987 to December 1994, 183 patients underwent a myocardial revascularization procedure with the use of at least two arterial grafts (IMAs, rGEA, IEA) at our institute. Their mean age was 56 +/- 8.7 years, the redo-operation rate was 16.9% (31/183), two-vessel disease was present in 61 patients (33.3%), three-vessel disease in 122 (66.7%). The LIMA was used in 179 patients (97.8%), the RIMA in 116 (63.4%), the rGEA in 66 (36.1%) and the IEA in 41 (22.4%). In-hospital mortality was 1.1% (2/183), while the perioperative myocardial infarction (MI) rate was 2.2% (4/183). The angiographic restudy, performed on 87 (47.5%) patients during the early postoperative period (median 38 days) showed the following grafts patency rates: LIMA 98.8 (86/87), RIMA 97.1 (34/35), IEA 85.7 (24/28), rGEA 97.05 (33/34) and saphenous vein 90.9% (10/11). The median follow-up was 35 months. Kaplan-Meier survival was 96 +/- 2% at 3 and 5 years, freedom from angina 94 +/- 2% at 3 years and 91 +/- 3% at 5 years, while the Kaplan-Meier freedom from cardiac events was 90 +/- 3% at 3 years and 88 +/- 3% at 5 years. Cox regression analysis identified perioperative MI (P = 0.03, relative risk 3.6) as the only prognostic factor for mortality at follow-up. With regards to recurrence of angina, multivariate analysis has shown that incremental risk factors for the return of angina are redo-operation (P < 0.01, relative risk 2.7) and the persistence of hypertension after surgery (P < 0.01; relative risk 3.2), while the use of the RIMA in the operation has emerged as a protective factor (P = 0.02; relative risk 0.43). Finally, only redo-operation (P < 0.01; relative risk 2.3), has emerged as a predictor of cardiac complications. Myocardial revascularization with at least two arterial grafts can be performed with very low perioperative morbidity and mortality and good mid-term follow-up. The mid-term results of arterial myocardial revascularization are less favourable in cases of redo-operations or when the RIMA is not used.


Determining competence of experienced critical care nurses.
A prescriptive algorithm to determine competence in an adult critical care setting guides cost-effective hiring and education of critical care nurses at the VA Medical Center in West Los Angeles. Outcomes included efficient mobilization of 15 RNs with critical care experience and a management tool that facilitated interviewing of candidates. Duplication of educational programs was avoided, thus saving this federal institution's inservice dollars while assuring quality care.


[Nutritional value and safety of homogenized quick-frozen mixtures, prepared from dagestan grown fruits and berries].
The mineral structure, the maintenance of vitamin C and phenolic substances in three-componental, homogeneous mixes from the fast freezing (-30 degrees C) fruits and berries are investigated. Experimental dates show, that the low-temperature freezing and the subsequent three- and nine-monthly storage (-18 degrees C) allow to keep in mixes quantity of the main things nutrients up to 70-80%. The developed technology of reception of fruit-berry mixes, on the basis of a principle of complementarity on the basic components of biochemical structure describing food value, can promote the decision of a problem of all-the-year-round supply of the population the rational, balanced feed.


Effect of exercise on the expression of adiponectin mRNA and GLUT4 mRNA in type 2 diabetic rats.
To investigate the impact of exercise on the expression of adiponectin and GLUT4 mRNA in type 2 diabetic rats, type 2 diabetic rat model was made. The diabetic rats were treated with swimming training for 8 weeks. The expression of adiponectin mRNA in perirenal fat and GLUT4 mRNA in skeletal muscles were assessed by reverse transcription polymerase chain reaction (RT-PCR) and the levels of blood glucose, serum insulin, and blood lipid were measured. Our results showed that the expression of adiponectin mRNA and GLUT4 mRNA in diabetic model group was decreased by 45% (P < 0.01), 43% (P < 0.01) respectively. The gene expression of adiponectin and GLUT4 was increased significantly in swimming group (P < 0.05 and P < 0.01, respectively). Compared with the model group, fasting insulin, TG, TC and FFA were decreased significantly in the training group (P < 0.05 or P < 0.01) as compared with model group. It is concluded that exercise can promote the expression of adiponectin mRNA and GLUT4 mRNA in type 2 diabetic rats, which may be one of the mechanisms responsible for the amelioration of insulin resistance in the rats.


The prognostic value of circulating myeloblasts in patients with myelodysplastic syndromes treated with azacitidine.


Enhancement (by native type II collagen) of the humoral immune response to native type I collagen in the rat.
Both native type II collagen and rabbit IgG were good immunogens in the rat when emulsified in incomplete Freund's adjuvant but native type I collagen and denatured type II collagen were not. Both native type II collagen and rabbit IgG had an enhancing effect upon the production of antibodies to native type I collagen when rats were immunised with a mixture of native type I collagen and either of these immunogens but denatured type II collagen did not have this effect. These immunogens, however, did not enhance the production of antibodies to ovalbumin when emulsified with it. Although both native type II collagen and rabbit IgG showed this enhancing effect with native type I collagen, only rats immunised with native type II collagen or mixtures containing native type II collagen developed arthritis. Thus, a specific immune response to collagen rather than this enhancing effect appears to be necessary for the induction of arthritis.


Dietary modulation of the biotransformation and genotoxicity of aflatoxin B(1).
Diet and its various components are consistently identified as among the most important 'risk factors' for cancer worldwide, yet great uncertainty remains regarding the relative contribution of nutritive (e.g., vitamins, calories) vs. non-nutritive (e.g., phytochemicals, fiber, contaminants) factors in both cancer induction and cancer prevention. Among the most potent known human dietary carcinogens is the mycotoxin, aflatoxin B(1) (AFB). AFB and related aflatoxins are produced as secondary metabolites by the molds Aspergillus flavus and Aspergillus parasiticus that commonly infect poorly stored foods including peanuts, pistachios, corn, and rice. AFB is a potent hepatocarcinogenic agent in numerous animal species, and has been implicated in the etiology of human hepatocellular carcinoma. Recent research has shown that many diet-derived factors have great potential to influence AFB biotransformation, and some efficiently protect from AFB-induced genotoxicity. One key mode of action for reducing AFB-induced carcinogenesis in experimental animals was shown to be the induction of detoxification enzymes such as certain glutathione-S-transferases that are regulated through the Keap1-Nrf2-ARE signaling pathway. Although initial studies utilized the dithiolthione drug, oltipraz, as a prototypical inducer of antioxidant response, dietary components such as suforaphane (SFN) are also effective inducers of this pathway in rodent models. However, human GSTs in general do not appear to be extensively induced by SFN, and GSTM1 - the only human GST with measurable catalytic activity toward aflatoxin B(1)-8,9-epoxide (AFBO; the genotoxic metabolite of AFB), does not appear to be induced by SFN, at least in human hepatocytes, even though its expression in human liver cells does appear to offer considerable protection against AFB-DNA damage. Although induction of detoxification pathways has served as the primary mechanistic focus of chemoprevention studies, protective effects of chemoprotective dietary components may also arise through a decrease in the rate of activation of AFB to AFBO. Dietary consumption of apiaceous vegetables inhibits CYP1A2 activity in humans, and it has been demonstrated that some compounds in those vegetables act as potent inhibitors of human CYP1A2 and cause reduced hCYP1A2-mediated mutagenicity of AFB. Other dietary compounds of different origin (e.g., constituents of brassica vegetables and hops) have been shown to modify expression of human hepatic enzymes involved in the oxidation of AFB. SFN has been shown to protect animals from AFB-induced tumors, to reduce AFB biomarkers in humans in vivo and to reduce efficiently AFB adduct formation in human hepatocytes, although it appears that this protective effect is the result of repression of human hepatic CYP3A4 expression, rather than induction of protective GSTs, at least in human hepatocytes. If this mechanism were to occur in vivo in humans, it would raise safety concerns for the use of SFN as a chemoprotective agent as it may have important implications for drug-drug interactions in humans. A dietary chemoprevention pathway that is independent of AFB biotransformation is represented by the potential for dietary components, such as chlorophyllin, to tightly bind to and reduce the bioavailability of aflatoxins. Chlorophyllin has been shown to significantly reduce genotoxic AFB biomarkers in humans, and it therefore holds promise as a practical means of reducing the incidence of AFB-induced liver cancer. Recent reports have demonstrated that DNA repair mechanisms are inducible in mammalian systems and some diet-derived compounds elevated significantly the gene expression of enzymes potentially involved in nucleotide excision repair of AFB-DNA adducts. However, these are initial observations and more research is needed to determine if dietary modulation of DNA repair is a safe and effective approach to chemoprevention of AFB-induced liver cancer.


Can the height-independent Pottel eGFR equation be used as a screening tool for chronic kidney disease in children?
Determination of plasma creatinine (Pcr) should be associated to an estimation of glomerular filtration rate (eGFR). Pottel et al. established a height-independent equation, eGFR = 107.3/(Pcr/Q) where Q is the median of Pcr (Pottel-Belgium). The aims were to 1) determine a local height-independent equation (Pottel-Lyon), 2) evaluate the performance of these equations compared to the Schwartz 2009 and Schwartz-Lyon equations, and 3) evaluate the height-independent equations in laboratory routine. Therefore, 1) all first pediatric Pcr determination (December 2009-June 2011) were collected, and median of Pcr was determined for each 1-year age interval (Q-Lyon), 2) GFR was measured (mGFR) in 359 children (438 measures) and compared to eGFR, and 3) all first Pcr determination (January 2012-June 2013) were used to calculate eGFR with the Pottel-Lyon and the Pottel-Belgium equations. Pcr was determined by an IDMS-standardized enzymatic assay. In the population with a mGFR, the Pottel-Lyon and the Schwartz-Lyon showed the best performance (bias, P10 and P30). However, the performance in identifying patients with a mGFR < 75 mL/min/1.73 m(2) was similar for all the studied equations. The performance of the height-independent and dependent equations to identify mild renal dysfunction is similar. The height-independent Pottel equation could be proposed as an excellent screening tool for kidney disease when height information is not available. " " • Determination of plasma creatinine in children is rarely associated to an estimation of glomerular filtration rate due to the lack of height information. • Pottel et al. developed a height-independent equation (eGFR = 107.3/(Pcr/Q) where Q is the median of Pcr for each age class. " " • The performance of the height-independent (Pottel) or height-dependent (Schwartz) equations is similar to identify renal dysfunction (GFR < 75 mL/min/1.73 m (2) ) in children. • The height-independent Pottel equation could be an excellent screening tool for kidney disease in a general pediatric laboratory when height information is not available.


Aging of the erythrocyte. IX. Fluorescence studies on changes in membrane properties.
Fluorescence studies of membranes prepared from density-separated red blood cells demonstrated an increase in lipid viscosity (judging from fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene and 1-aminonaphthalene-8-sulfonate and changes in the physical state of proteins (judging from tryptophan fluorescence intensity and polarization). These alterations may be partly due to lipid peroxidation. In vivo accumulation of products of this process in the membranes was confirmed. Changes were also found in the distribution pattern of cell populations of different mean age in a cell sorter, probably due to alterations in rheological properties of the erythrocytes.


In vitro testing of endothelial cell monolayers under dynamic conditions inside a beating ventricular prosthesis.
Thromboembolic complications remain a major problem associated with the long-term clinical use of cardiac prostheses. A promising approach toward resolving this predicament is lining the blood contacting surfaces with a functional monolayer of endothelial cells (EC). In developing an endothelialized cardiac prosthesis, the authors in the past focused on establishing a confluent EC monolayer on the luminal surface of ventricular blood sacs. In this study, the authors concentrated on exposing the post confluent monolayers to the dynamic conditions inside a beating ventricle. The cells, derived from either bovine aortae or jugular veins, were grown to post confluence inside fully assembled ventricles on fibronectin or plasma cryoprecipitate coated, textured surfaces. After 11 days of culturing under static conditions, the endothelialized ventricles were connected to a mock loop that was run for 6 and 24 hr at 60 bpm and mean flow rate of 3.2 L/min. The status of the monolayer was evaluated by Alamar Blue assay before and after each run, and the extent of surface coverage was determined visually using bright field microscopic study after cell staining with KMnO4 and toluidine blue. In addition, morphometric information on cells/polyurethane surface was obtained with a scanning electron microscope. After 6 hr of pumping, cell staining revealed signs of moderate cell loss in fibronectin coated blood sacs, whereas in cryoprecipitate coated bladders the signs of denudation were marginal. In seven ventricles operated for 24 hr, Alamar Blue measurements indicated 35 +/- 16% of cell loss from monolayers established on fibronectin coating, but only 4.8 +/- 6.25% on cryoprecipitate. Thus, the current study demonstrates the feasibility of maintaining an intact endothelial surface in a beating ventricular prosthesis and indicates that the integrity of the endothelial lining is dependent upon a proper choice of surface macrostructure and protein coating.


Pathological fracture after migration of cement used to treat distal femur physeal arrest.
Treatment of physeal arrest with bar removal and placement of interposition materials in young patients has been shown to restore physeal growth. Among the various materials that have been used to prevent early reformation of the physeal bar (fat, silastic, cartilage), Peterson recommended the use of Cranioplast, as this material can prevent bar reformation, and it is radiolucent because it does not contain barium. Peterson suggested steps be taken to prevent migration of the Cranioplast, as migration of the interposition material might allow for reformation of the physeal bar. Although Peterson had not observed cement migration leading to pathologic fracture, he felt that this was a potential concern. We describe the case of cement migration from the epiphysis into the diaphysis, leading to a pathological femur fracture. Despite migration of the Cranioplast, the physeal bridge did not reform, and the patient had nearly normal growth 7 years after the initial physeal arrest procedure.


Atrial fibrillation and congestive heart failure: a two-way street.
Atrial fibrillation (AF) is the most common sustained cardiac arrhythmia encountered in clinical cardiology, particularly in patients with heart failure. Its prevalence increases with the degree of left ventricular dysfunction and severity of heart failure symptoms. The development of AF in the setting of heart failure has been shown to result in increased mortality. Studies thus far indicate that rhythm control with antiarrhythmic drug therapy or catheter ablation offers both symptomatic and probably survival advantage in patients with heart failure and AF. In patients with permanent AF, the effects of restoring a regular ventricular response with atrioventricular junction ablation followed by biventricular pacing remain to be shown. In the current manuscript, we will review the proposed mechanisms of increased morbidity and mortality associated with AF and the current treatment options including the roles of radiofrequency ablation and pacing.


In vivo kinetics of trichloroacetate in male Fischer 344 rats.
Trichloroacetate (TCA) is a toxicologically important metabolite of the industrial solvents trichloroethylene and tetrachloroethylene, and a by-product of the chlorination of drinking water. Tissue disposition and elimination of 14C-TCA were investigated in male Fischer 344 rats injected iv with 6.1, 61, or 306 micromol TCA/kg body weight. Blood and tissues were collected at various time points up to 24 h. No metabolites were observed in plasma, urine, or tissue extracts. Overall TCA kinetics in tissues were similar at all doses. Based on similar terminal elimination rate constants, tissues could be divided into three classes: plasma, RBC, muscle, and fat; kidney and skin; and liver, small intestine, and large intestine. Nonextractable radiolabel, assumed to be biologically incorporated metabolites in both liver and plasma, increased with time, peaking at 6-9 h postinjection. The fraction of the initial dose excreted in the urine at 24 h increased from 67% to 84% as the dose increased, whereas fecal excretion decreased from 7% to 4%. The cumulative elimination of TCA as CO2 at 24 h decreased from 12% to 8% of the total dose. Two important kinetic processes were identified: a) hepatic intracellular concentrations of TCA were significantly greater than free plasma concentrations, indicating concentrative transport at the hepatic sinusoidal plasma membrane, and b) TCA appears to be reabsorbed from urine postfiltration at the glomerulus, either in the renal tubules or in the bladder. These processes have an impact on the effective tissue dosimetry in liver and kidney and may play an important role in TCA toxicity.


Stimulation of anaerobic metabolism in rats at high altitude hypoxia--adrenergic effects dependent on dietary states.
1. Plasma lactate and pyruvate were increased more markedly in fed rats than in fasted rats exposed to an 8000 m altitude. 2. The increase in plasma lactate and pyruvate was enhanced and inhibited by the alpha 1-adrenergic antagonist prazosin and the beta-blocker propranolol, respectively, in fasted rats exposed to an 8000 m altitude. Blood glucose was not changed by adrenergic blockades under the same conditions. 3. Prazosin and propranolol showed no effect on glycolytic metabolites in plasma in fed rats submitted to an 8000 m altitude. Blood glucose of fed rats was increased by alpha 1-blockade during severe hypoxia. 4. In fasted rats whose energy metabolism depends on oxidation mainly, alpha 1- and beta-adrenergic receptors can participate in the stimulation of respiration and the glycogen degradation, respectively, during an exposure to severe hypoxia. In fed rats energy metabolism depends on glycolysis, which utilizes blood glucose as the substrate preferentially during hypoxia.


[Bilateral recurrent retinal thrombosis in a young man].
A 41-year-old male patient presented with veiled vision which had appeared only a few hours previously. Funduscopy revealed a retinal edema due to venous stasis retinopathy. A previous history was unknown except for an uncorrected arterial hypertension. A retinal vein thrombosis and macula edema developed in the affected eye. An antiphospholipid antibody syndrome was diagnosed which was treated with anticoagulants.


LDL apheresis for cholesterol embolism following coronary artery bypass graft surgery--a case report.
A 76-year-old man without any prior history of abnormal urinalysis findings or renal insufficiency demonstrated mild renal dysfunction after coronary bypass graft surgery (CABG). Two months after CABG, pain and blueness in the toes (blue toe syndrome) appeared and, the serum creatinine level (S-Cr) increased from 1.2 to 2.0 mg/dL. On admission (3 months later), the urinary protein level was 0.5 g/day, white blood cell count 8,300/microL with eosinophils (Eo) 10.5%, S-Cr 2.1 mg/dL, and low-density lipoprotein (LDL) 106 mg/dL. Acute renal failure and blue toe syndrome due to a cholesterol embolism (CE) were diagnosed. Alprostadil 40 microg/day orally for 2 weeks and alprostadil 40 microg/day intravenously were used for 5 weeks, and Eo were 250/microL, S-Cr 2.5 mg/dL; however, blue toe syndrome gradually developed. At 8 weeks after admission, limaprost alfadex 30 microg/day orally was used for 3 weeks. However, the Eo gradually rose to 1,520/microL, S-Cr to 3.0 mg/dL, and LDL to 135 mg/dL, and LDL apheresis was therefore performed 20 times for CE. The data just after LDL apheresis was performed 10 times were as follows: Eo 1,120/microL, S-Cr 4.0 mg/dL, and LDL 89 mg/dL, and blue toe syndrome had disappeared. At 10 months after the first LDL apheresis, the Eo were 630/microL, S-Cr 2.9 mg/dL, and LDL 109 mg/dL. As a result, LDL apheresis was found to be beneficial for the treatment of CE with acute renal failure and blue toe syndrome after CABG.


[A case of patent urachus].
A case of congenital patent urachus is reported. A 12-month-old boy was referred to our outpatient clinic with the complaint of watery discharge from the navel. A cystography revealed the communication between the dome of the urinary bladder and the umbilicus. Excretion from the umbilicus of indigocarmine solution instilled into the urinary bladder was recognized. There were no complicated abnormalities in other organs. The patient underwent radical operation. Herein, we collected 164 cases of urachal disorders reported in Japan and reviewed the incidence, clinical symptoms, diagnosis and treatment. The classifications of the disease were briefly discussed.


Course and prognosis of chronic progressive multiple sclerosis. Results of an epidemiological study.
In studies on the natural course of multiple sclerosis (MS), several forms of the disease are distinguished. The most important are the relapsing remitting and the chronic progressive forms. The relationship between these remains unclear. In a prospective epidemiological survey we studied the course of MS using the year in which the chronic-progressive phase started as a landmark. The reliability of this "year of progression" was examined in an observer agreement study. Data were acquired from 342 patients. Progression of the handicap was most rapid in case of a secondary progressive course, female sex, high relapse rate in the preceding remitting phase and "year of progression" at a higher age. Survival after the "year of progression" was lowest in the secondary progressive group. Determining the "year of progression" seems to be significant for the prognosis.


Stimulus-specific regulation of CD63 and CD203c membrane expression in human basophils by the flavonoid quercetin.
Flavonoids, such as quercetin, were reported to inhibit histamine release and cytokine production by basophils, but there is no evidence describing their action on membrane markers and intracellular biochemical pathways. The aim of the study was to examine the effect of several quercetin doses on an in vitro human basophil activation system that evaluates up-regulation of membrane markers in response to agonists. Leukocyte buffy coats from K(2)-EDTA anti-coagulated blood were treated with different concentrations of quercetin and triggered with anti-IgE ("allergy model") and with N-formyl-Met-Leu-Phe (fMLP) ("inflammation model"). Basophils were captured as CD123(bright)/HLA-DR(non-expressing) cells in a flow cytometry analysis and fluorescence values of CD63-FITC, CD203c-PE and CD123-PECy5 were used to produce dose response curves. Quercetin at a dose of 10 microg/ml strongly inhibited CD63 and CD203c membrane up-regulation triggered by both agonists, but it neither affected cell viability nor changed the expression of the phenotypic marker CD123. The anti-IgE model appeared highly sensitive to the effect of quercetin: a dose as low as 0.01 microg/ml was able to significantly decrease CD63 and CD203c membrane expression. In the fMLP model the dose response was different: quercetin doses from 0.01 to 0.1 microg/ml significantly increased up-regulation of membrane markers, achieving the highest effect with CD63. Very low doses of quercetin, within the pharmacological range, inhibit IgE-mediated membrane marker's up-regulation but prime the response to the chemotactic peptide fMLP; this stimulus specificity may have implications on the possible therapeutic action of the flavonoid in different pathologies.


Modulation of enantioselective metabolism and inversion of ibuprofen by xenobiotics in isolated rat hepatocytes.
R-ibuprofen undergoes chiral inversion by the formation of a coenzyme A (CoA) thioester and subsequent epimerization and hydrolysis. Using isolated rat hepatocytes, the interaction of xenobiotics with the inversion and oxidation pathways of ibuprofen enantiomers was determined from the time course of R- and S-ibuprofen and ibuprofenyl-CoA during 4-hr incubations with R- or S-ibuprofen (25 microM). By fitting a first-order model, the rate constants of the formation of ibuprofenyl-CoA (K12), oxidation of R-ibuprofen (K10), hydrolysis of ibuprofenyl-CoA (K21) and oxidation of S-ibuprofen (K30) were 1.306, 0.284, 6.858 and 0.496 hr-1, respectively. The fractional inversion of R-ibuprofen was 0.75 and the area under the curve for ibuprofenyl-CoA was 203.8 microM min. Coincubation with 50 microM of the cytochrome P450 inhibitors metyrapone and proadifen resulted in significant reductions of K10 and K30; the fractional inversion of R-ibuprofen increased to 116% and 127% and the area under the curve of ibuprofenyl-CoA to 145% and 144% of controls, respectively. Valproic acid and pivalic acid at 50 microM significantly reduced the K12 and increased the K21; the fractional inversion was unchanged but the area under the curve of ibuprofenyl-CoA was significantly reduced to 57% and 28% of controls, respectively. Valproic acid also significantly reduced K10 and K30. p-nitrobenzoic acid at 50 microM significantly increased K21 and reduced the area under the curve of ibuprofenyl-CoA to 44% of control but did not influence the fractional inversion. Selective inhibitors of ibuprofen oxidation were found to enhance significantly hepatocellular exposure to the potentially reactive ibuprofenyl-CoA intermediate.


Effectiveness of a vacuum lifting system in reducing spinal load during airline baggage handling.
Information on spinal loading for using lift assist systems for airport baggage handling is lacking. We conducted a laboratory study to evaluate a vacuum lift system for reducing lumbar spinal loads during baggage loading/unloading tasks. Ten subjects performed the tasks using the industry average baggage weight of 14.5 kg on a typical two-shelved baggage cart with or without using the lift system (i.e. lifting technique). Repeated measures analysis of variance (2 tasks × 2 shelf heights x 2 techniques) was used. Spinal loads were estimated by an electromyography-driven biomechanical model. On average, the vacuum lift system reduced spinal compressive forces on the lumbar spine by 39% and below the 3400 N damage threshold. The system also resulted in a 25% reduction in the anterior-posterior shear force at the L5/S1 inferior endplate level. This study provides evidence for the potential to reduce spinal loads when using a vacuum lift system.


Professional care providers in dementia care in eight European countries; their training and involvement in early dementia stage and in home care.
Knowledge concerning professionals involved in dementia care throughout its trajectory is sparse; the focus has mainly been on nursing-home care and less on home care, diagnosis and treatment of the disease and its complications despite the fact that home care is the most prominent type of care. The aim of this study was to explore and describe professional care providers involved in dementia care and their educational level applying the International Standard Classification of Education (ISCED) and further to investigate practice in the RightTimePlaceCare-countries with regard to screening, diagnostic procedures and treatment of dementia and home care. The findings demonstrate more similarities than differences in terms of type of professionals involved among the countries although untrained staff were more common in some countries. Findings also show that many types of professionals are involved, who to turn to may not be clear, for instance in terms of medical specialities and it may be unclear who bears the ultimate responsibility. The professionals involved in diagnosis, treatment and care are educated to bachelor's level or above whilst everyday care is provided by people trained at a lower ISCED level or with no formal training. Registered nurses as well as occupational therapists have bachelor's degrees in most countries, but not in Germany or Estonia. Professionals specifically trained in dementia care are not so common. Further research is needed to reveal not only who provides the diagnostics and treatment, but also how home care is organised and quality assured. Many different types of professionals serve as providers along the trajectory of the disease which may be difficult for the patient and the informal caregiver to cope with.


Shear Wave Elastography in Rectal Cancer Staging, Compared with Endorectal Ultrasonography and Magnetic Resonance Imaging.
The goal of the study described here was to investigate the value of shear wave elastography (SWE) in pre-operative staging of rectal cancer. Fifty-five patients with rectal cancer underwent pre-operative conventional endorectal ultrasonography (ERUS), SWE and enhanced magnetic resonance imaging (MRI) examinations. Pathologic results were used as the gold standard for cancer staging. The concordance rate with pathologic stage by ERUS and MRI and the stiffness values measured by SWE for tumors in different stages were compared. The concordance rates for cancer staging were 72.7% and 70.9% for conventional ERUS and enhanced MRI, respectively; the difference was not significant (p > 0.05). SWE indicated that the mean and maximum stiffness values of the tumors increased with advance in stage. The differences in stiffness values between T1 and T2, T1 and T3-4, as well as T2 and T3-4, were all statistically significant (p < 0.001). When the maximum stiffness values of 65.0 and 90.7 kPa are used for the diagnosis of T1, T2 and local advanced rectal cancer, the concordance rate of cancer staging was 85.5%, which was slightly higher than those of ERUS and MRI, although the difference was not statistically significant (p > 0.05). SWE is useful in judging the depth of invasion of rectal tumors. The value of tumor stiffness can provide a quantifiable indicator for pre-operative diagnosis of cancer staging and can be used as a supplement to conventional ERUS. Further studies with larger sample sizes are needed.


Preconditioning by hypoventilation increases ventricular arrhythmia threshold in Wistar rats.
Hypoventilation, as one of ventilatory disorders, decreases the electrical stability of the heart similarly as ischemia. If preconditioning by short cycles of ischemia has a cardioprotective effect against harmful influences of a prolonged ischemic period, then preconditioning by hypoventilation (HPC) can also have a similar effect. Anesthetized rats (ketamine 100 mg/kg + xylasine 15 mg/kg i.m., open chest experiments) were subjected to 20 min of hypoventilation followed by 20 min of reoxygenation (control group). The preconditioning (PC) was induced by one (1PC), two (2PC) or three (3PC) cycles of 5-min hypoventilation followed by 5-min reoxygenation. The electrical stability of the heart was measured by a ventricular arrhythmia threshold (VAT) tested by electrical stimulation of the right ventricle. Twenty-minute hypoventilation significantly decreased the VAT in the control and 1PC groups (p<0.05) and non-significantly in 2PC vs. the initial values. Reoxygenation reversed the VAT values to the initial level only in the control group. In 3PC, the VAT was increased from 2.32+/-0.69 mA to 4.25+/-1.31 mA. during hypoventilation (p<0.001) and to 4.37+/-1.99 mA during reoxygenation (p<0.001). It is concluded that cardioprotection against the hypoventilation/ reoxygenation-induced decrease of VAT proved to be effective only after three cycles of HPC.


Lingual calcinosis circumscripta in a captive sitatunga.
Calcinosis circumscripta (CC) was found in a 10-yr-old female sitatunga (Bovidae; Tragelaphus spekei). At necropsy, there were two white coalescing nodules (3 x 5 x 2 cm, 2 x 2 x 1.5 cm) on the ventral side of the tongue. The cut surface of the nodules had multiple, well-circumscribed loculi with chalky appearance and gritty consistency, separated by thin strands of connective tissue. Histologically, the nodules contained multiple loculi of various sizes and shapes, which consisted of amorphous material that was pale basophilic with H&E stain, surrounded by fibrous connective tissue. Around the loculi were numerous foreign body giant cells and epithelioid macrophages; occasional lymphocyte aggregations also were seen. The amorphous material was positive for PAS and von Kossa's stain. Calcinosis circumscripta is rare in the Bovidae.


Features of the peritoneal covering of the lesser pelvis with special reference to stomata regions.
Occasional reports describe various aspects of the fine morphology of the pelvic peritoneum, but its complete organ characteristics remain undefined. The peritoneal covering of the urinary bladder, rectum, uterus, uterine tube, ovary, broad ligament (BL) and testis in Wistar rats was examined by means of transmission and scanning electron microscopy (TEM, SEM). Unusually complicated relief and stomata between the cubic mesothelial cells characterized the surface of the BL. Deep, parallel furrows separated the wide longitudinal folds over the entire length of the uterine tube. The uterus and the ovary formed less numerous, shallow or extremely deep crypt-like invaginations, as well as serous villus-like or papilla-like evaginations. The flat cells were the predominant cell type over the BL, while the cubic mesothelium was the basic covering of the organs. Most of the cubic cells were located in the invagination of the submesothelial layer (SML). Such cells formed an almost smooth surface over the urinary bladder or formed larger areas of the rectum and the testis surfaces. Numerous microvilli, ciliae, round evaginations and complex lamellar bodies characterized their apical plasmalemma. In conclusion, the mesothelial heterogeneity is a stable feature of the lesser pelvis peritoneum, confirmed by TEM and SEM. The cubic mesothelium characterizes the organ peritoneum, while the BL plays the role of the parietal sheet, involving lymphatic units in the SML. The different types of contacts between the mesothelio-endothelial cells, large lymphatic vessels and occasional stomata are the usual components of the lymphatic units in norm, visible by TEM. Images of stomata, seen by SEM, demonstrate oval-shaped deep channel-like gaps surrounded by cubic mesothelium. The last data extend the evidence on stomata regions, which resemble the diaphragmatic ones. Clusters of cells (macrophages, mastocytes and Lymphocytes), small vessels (blood or lymphatic) and nerve fibers (unmyelinated and rare myelinated) form highly specialized complexes in the SML of the ovary, the uterus and the testis.


Increased serum levels of progranulin (PGRN) in patients with haemophilic arthropathy.
Haemophilia A and B are rarely occurring X chromosome-linked congenital coagulation disorders dominated by spontaneous joint bleedings and chronic synovitis, leading to development of haemophilic arthropathy (HA). Progranulin (PGRN) is a growth factor with anti-inflammatory and immunomodulatory properties. PGRN is an important molecule in the pathogenesis of osteoarthritis (OA) and rheumatological disorders. This study was aimed at investigating the potential role of PGRN in the mechanisms underlying the pathogenesis of HA. The serum levels of PGRN were measured by enzyme-linked immunosorbent assay (ELISA) in patients with end-stage knee joint HA (n = 20) and end-stage primary knee joint OA (n = 20) who met the inclusion and exclusion criteria. The clinical and radiological assessment of disease severity was evaluated by the Knee Society Score (KSS) and Kellgren-Lawrence scale. Median PGRN levels in HA patients was 349.1 ng/mL (232.8-415.6 ng/mL) and in OA patients 148.3 ng/mL (112.1-275.3 ng/mL) with statistically significant differences between both groups (P < 0.015). Further analysis revealed no correlation between PGRN levels and any of the patient demographics and clinical parameters. This study demonstrates increased PGRN serum levels in patients with HA and provides new insights into the mechanisms underlying the pathogenesis of HA indicating a new potential target for therapeutic intervention.


Polyamines modulate streptomycin-induced mistranslation in Escherichia coli.
The effects of intracellular levels of polyamines on both the in vivo inhibition of protein synthesis and the decrease of translation accuracy induced by streptomycin have been studied in polyamine-auxotrophic strains of Escherichia coli infected with the MS2 bacteriophage. The amount of viral coat protein formed was strongly reduced upon addition of increasing concentrations of streptomycin to polyamine-supplemented bacteria. In contrast, the antibiotic almost did not inhibit coat protein synthesis in polyamine-starved cells. The increase of mistranslation frequency elicited by streptomycin was only observed in bacteria grown with putrescine. In these cells several coat protein-satellites were detected after two-dimensional gel electrophoresis. These proteins, more basic than the normal MS2 coat protein, contain multiple substitutions of lysine for asparagine.


Correlation of quality of life and functional outcome measures for cervical spondylotic myelopathy.
Studies comparing surgical treatments for cervical spondylotic myelopathy (CSM) are heterogeneous, using a variety of different quality of life (QOL) outcomes and myelopathy-specific measures. This study sought to evaluate the relationship of these measures to each other, and to better understand their use in evaluating patients with CSM. A retrospective study was performed in all patients with CSM who underwent either ventral or dorsal cervical spine surgery at a single tertiary-care institution between January 2008 and July 2013. Severity of myelopathy was assessed pre- and postoperatively using both the Nurick scale and the modified Japanese Orthopaedic Association (mJOA) classification of disability. Prospectively collected QOL outcomes data included Pain Disability Questionnaire (PDQ), Patient Health Questionnaire-9 (PHQ-9), and EQ-5D. Spearman rank correlations were calculated to assess the construct convergent validity for each pair of health status measures (HSMs). To assess each HSM's ability to discriminate favorable EQ-5D index, we performed receiver operating characteristic (ROC) curve analysis and assessed the area under the curve (AUC). A total of 119 patients were included. The PDQ total score had the highest correlation with EQ-5D index (Spearman's rho = -0.82). Neither of the myelopathy scales (mJOA or Nurick) had strong correlations between themselves (0.41) or with the other QOL measures (absolute value range 0.13-0.49). In contrast, the QOL measures correlated relatively well with each other (absolute value range 0.68-0.97). For predicting favorable EQ-5D outcomes, PDQ total score had an AUC of 0.909. The AUCs were significantly greater for the QOL measures in comparison with the myelopathy measures (AUCs were 0.677 and 0.607 for mJOA and Nurick scale scores, respectively). The authors found that all included measures of QOL and CSM-specific (mJOA or Nurick scale) measures were valid and responsive. The PDQ was the most predictive of positive QOL after surgery (as measured by the EQ-5D index) for patients with CSM. The substantially lower correlation between myelopathy and QOL outcomes, compared with the various QOL measures themselves, suggests that these questionnaires are measuring different aspects of the patient experience. Solely assessing the myelopathy or disease-specific signs and symptoms is likely insufficient to fully understand and appreciate clinical outcome in its totality. These questionnaire types should be used together to best evaluate patients pre- and postoperatively.


Test of Gross Motor Development-3 (TGMD-3) with the Use of Visual Supports for Children with Autism Spectrum Disorder: Validity and Reliability.
The validity and reliability of the Test of Gross Motor Development-3 (TGMD-3) were measured, taking into consideration the preference for visual learning of children with autism spectrum disorder (ASD). The TGMD-3 was administered to 14 children with ASD (4-10 years) and 21 age-matched typically developing children under two conditions: TGMD-3 traditional protocol, and TGMD-3 visual support protocol. Excellent levels of internal consistency, test-retest, interrater and intrarater reliability were achieved for the TGMD-3 visual support protocol. TGMD-3 raw scores of children with ASD were significantly lower than typically developing peers, however, significantly improved using the TGMD-3 visual support protocol. This demonstrates that the TGMD-3 visual support protocol is a valid and reliable assessment of gross motor performance for children with ASD.


Tula hantavirus L protein is a 250 kDa perinuclear membrane-associated protein.
The complete open reading frame of Tula hantavirus (TULV) L RNA was cloned in three parts. The middle third (nt 2191-4344) could be expressed in E. coli and was used to immunize rabbits. The resultant antiserum was then used to immunoblot concentrated TULV and infected Vero E6 cells. The L protein of a hantavirus was detected, for the first time, in infected cells and was found to be expressed as a single protein with an apparent molecular mass of 250 kDa in both virions and infected cells. Using the antiserum, the expression level of the L protein was followed and image analysis of immunoblots indicated that there were 10(4) copies per cell at the peak level of expression. The antiserum was also used to detect the L protein in cell fractionation studies. In cells infected with TULV and cells expressing recombinant L, the protein pelleted with the microsomal membrane fraction. The membrane association was confirmed with membrane flotation assays. To visualize L protein localization in cells, a fusion protein of L and enhanced green fluorescent protein, L-EGFP, was expressed in Vero E6 cells with a plasmid-driven T7 expression system. L-EGFP localized in the perinuclear region where it had partial co-localization with the Golgi matrix protein GM130 and the TULV nucleocapsid protein.


Antifungal susceptibility, exoenzyme production and cytotoxicity of novel oximes against Candida.
Novel oximes were synthesized, their in vitro antifungal activity against Candida was evaluated and their cytotoxicity was determined. The procedure used for the synthesis of the oximes is aligned with the current green chemistry trend; water is employed as the solvent in this reaction. The minimum inhibitory and minimum fungicidal concentrations of the oximes were evaluated using the CLSI M27-A3 method. The influence of these compounds on the inhibition of the production of hydrolytic enzymes, phospholipase and proteinase by Candida was also investigated. The compounds showed a good ability to inhibit phospholipase, with a 50 % reduction in most cases. However, the tested compounds did not affect proteinase. The current results showed a substantial reduction in the phospholipase production, which suggests that compounds of this class may interfere with host infection and disease progression. The oximes examined showed lower fungicidal activities than fluconazole but interfered significantly with the expression of phospholipase. Some of the oximes included in this study could be a suitable matrix for the development of novel antifungal compounds.


Intrathecal cholecystokinin interacts with morphine but not substance P in modulating the nociceptive flexion reflex in the rat.
The effect of intrathecal (IT) cholecystokinin (CCK), substance P (SP) and morphine (MO) on spinal cord excitability was studied in decerebrate, spinalized rats. CCK had a weaker facilitatory effect on the nociceptive flexion reflex than SP. The possible functional significance of the coexistence of CCK and SP in neurons projecting to the spinal cord was tested by coadministration of the two peptides. At the doses tested no synergistic interaction on the reflex was found with CCK and SP. IT MO caused a brief enhancement followed by a prolonged depression of the reflex. A high dose of CCK injected prior to MO increased the facilitatory effect and decreased the depressive effect of the opiate on the reflex. The effect of desulfated (D) CCK was similar to CCK but at a higher dose. Naloxone (NAL) had a similar effect as CCK when administered prior to MO. The MO-induced depression of the reflex was readily reversed by NAL, but not by CCK. The results indicate that CCK may prevent the inhibitory effect of MO on spinal cord excitability to nociceptive stimulation, but does not reverse it. CCK may alter the balance of excitation-inhibition between various types of dorsal horn interneurons that are involved in the transmission of nociceptive information.


The competitive success of Methanomicrococcus blatticola, a dominant methylotrophic methanogen in the cockroach hindgut, is supported by high substrate affinities and favorable thermodynamics.
Methanomicrococcus blatticola is an obligately anaerobic methanogen that derives the energy for growth exclusively from the reduction of methylated compounds to methane with molecular hydrogen as energy source. Competition for methanol (concentration below 10 microM) and H(2) (concentration below 500 Pa), as well as oxidative stress due to the presence of oxygen are likely to occur in the peripheral region of the cockroach hindgut, the species' normal habitat. We investigated the ecophysiological properties of M. blatticola to explain how it can successfully compete for its methanogenic substrates. The organism showed affinities for methanol (K(m)=5 microM; threshold<1 microM) and hydrogen (K(m)=200 Pa; threshold <0.7 Pa) that are superior to other methylotrophic methanogens (Methanosphaera stadtmanae, Methanosarcina barkeri) investigated here. Thermodynamic considerations indicated that 'methanol respiration', i.e. the use of methanol as the terminal electron acceptor, represents an attractive mode of energy generation, especially at low hydrogen concentrations. Methanomicrococcus blatticola exploits the opportunities by specific growth rates (>0.2 h(-1)) and specific growth yields (up to 7 g of dry cells per mole of methane formed) that are particularly high within the realm of mesophilic methanogens. Upon oxygen exposure, part of the metabolic activity may be diverted into oxygen removal, thus establishing appropriate anaerobic conditions for survival and growth.


Khorasan stroke registry: analysis of 1392 stroke patients.
The hospital-based stroke registry is useful for understanding diverse clinical characteristics of stroke related to geographical, racial, or environmental differences. The Khorasan Stroke Registry was established for evaluation of incidence, clinical manifestations, risk factors, topography, and etiology of ischemic stroke in Southern Khorasan, Iran, during 2001 - 2005. Consecutive stroke patients underwent a standard battery of diagnostic investigations by a stroke neurologist. Topography and etiology of brain infarction was determined based on the Practical Iranian Criteria classification. The incidence of ischemic stroke in Khorasan population is 43.17 cases per 100,000 people per year. During a 5-year period 1,392 ischemic stroke patients (738 females, 654 males) were evaluated in the Khorasan Stroke Registry. Atherosclerosis constituted 53.6% of etiologies followed by uncertain causes (19.9%), cardioembolism (11.8%), and miscellaneous etiologies (2.9%). Eleven point seven percent of our patients had both atherosclerosis and cardioembolic mechanisms. Rheumatic valvular disease was present in 44.8% of cardioembolic strokes and caused 4.31 preventable stroke cases per 100,000 Iranian population per year. Hypertension and history of ischemic cerebrovascular events were the most frequent risk factors, 53.1% and 22.3% respectively. In-hospital mortality of our ischemic stroke patients was 7.3%. High frequency of atherosclerotic etiology in the Khorasan Stroke Registry is because of its classification criteria, which does not separate small vessel territory infarcts as a different etiologic subtype. Rheumatic valvular disease is an important cause of stroke in Khorasan population.


Olive oil and mitochondrial oxidative stress.
This review summarizes studies on the role of olive oil intake in the prevention and attenuation of mitochondrial oxidative stress derived from several situations such as ageing, physical exercise, treatment with adriamycin or intake of thermally oxidised oils. After more than fifteen years of studies on these issues, results have demonstrated that under different models of oxidative stress the regular intake of virgin olive oil as dietary fat is able to attenuate or increase free radical production at the mitochondrial level to a lower extent than when n-6 polyunsaturated oils are used. In the same way, virgin olive oil leads to better function of the mitochondrial electron transport chain.


Prevalence of antibodies to seven viruses in a flock of ewes in Minnesota.
Blood samples were collected from a flock of healthy ewes at a University of Minnesota research station. Sera from these blood samples were tested for antibodies against 7 viruses, using 3 tests (eg, virus-neutralization test for bovine viral diarrhea virus, infectious bovine rhinotracheitis virus, bovine adenovirus type 3, and bovine respiratory syncytial virus; hemagglutination inhibition test for parainfluenza virus type 3; and agar-gel immunodiffusion test for lentivirus of ovine progressive interstitial pneumonia and bluetongue virus). The number of seropositive ewes for each antibody type were 1 of 377 (0.3%) for bovine viral diarrhea virus, 2 of 377 (0.5%) for infectious bovine rhinotracheitis virus, 29 of 378 (7.6%) for bovine adenovirus type 3, 200 of 378 (52.5%) for bovine respiratory syncytial virus, 273 of 373 (71.7%) for parainfluenza virus type 3, and 210 of 379 (55%) for ovine progressive pneumonia virus. All ewes were seronegative for bluetongue virus antibodies.


Spinal cord injury hospitalisation in a rehabilitation hospital in Japan.
The Kanagawa Rehabilitation Hospital was established by Kanagawa Prefecture 20 years ago. The hospital has a 40 bed SCI unit and has been providing comprehensive medical and rehabilitation services for SCI patients of this prefecture. A letter of enquiry to 404 major surgical units within this prefecture revealed that 630 SCI patients had been under care in this prefecture during a period of 3 years. Of these, the Frankel grade A, B and C patients, were assumed to be rehabilitation candidates, totalling about 130 in this prefecture per year. The inpatient medical record revealed that we treated 770 SCI patients from 1986 to 1991. From this data, we estimated that the hospital accepted an average of 54.5 SCI patients per year for rehabilitation purposes. Other inpatients were rehospitalised for various complications. According to the study of 125 patients who were hospitalised for comprehensive rehabilitation, they waited an average of 139 days and stayed in the hospital for 288 days. The increasing number of chronic patients who required treatment of complications appeared to be delaying the early admission of rehabilitation patients. The medical treatment cost for those with spinal cord injuries was less than that for the average patient with other types of medical conditions.


Vitamin D3 fortification, quantification, and long-term stability in Cheddar and low-fat cheeses.
Considering the widespread insufficiency of vitamin D, the fortification of additional foods with vitamin D is warranted. The objective of this research was to assess the feasibility of vitamin D3 fortification in natural hard cheeses. We examined the recovery, distribution, long-term retention, and heat stability of the vitamin in industrially made fortified Cheddar and low-fat cheeses. The results indicated that the vitamin D3 did not degrade during processing, over 1 year of ripening (3-8 degrees C), or after thermal treatment at 232 degrees C for 5 min. Vitamin D3 recovery in the fortified Cheddar and low-fat cheeses were, respectively, 91 and 55% of the vitamin D3 added to the milk used to make each cheese. The remaining vitamin D3 was entrained in the whey. The vitamin D3 was uniformly distributed throughout the blocks of cheese. The fortification process did not alter the yield, chemical composition, or flavor of the Cheddar cheese. We conclude that industrially manufactured Cheddar and low-fat cheeses are suitable for vitamin D3 fortification.