[The effect of sedation on oxygen uptake during spontaneous breathing].
Sedation may be used in intensive care and emergency medicine to improve the oxygen demand/delivery ratio. The influence of sedation has most frequently been investigated in a dose-related manner. The aim of the present study was to determine the effect-related influence of different sedatives on oxygen uptake (VO2) in relation to defined resting conditions. METHODS. Forty ASA I patients who had to undergo a minor surgical procedure were investigated 1.5 h before surgery at basal energy-expenditure measurement conditions. One of the following substances was given with a preset bolus rate in a double-blind, randomised order until a defined level of sleep or side effects was encountered: propofol (n = 8), midazolam (n = 8), thiopentone (n = 8), sodium chloride (n = 8), and fentanyl (n = 8). The sleep level was defined as sluggish response to a loud voice or tapping on the forearm. The variables VO2, carbon dioxide elimination (VCO2), end tidal CO2 (p(et)CO2), oxygen saturation (SaO2), heart rate, systemic blood pressure, skin temperature, and skin resistance on the sole of the foot were documented on-line on a computer. All variables were compared using differences of averages from 10-min periods before and after sedation during which the VO2 was minimal. RESULTS. The mean VO2 before sedation was 264 +/- 60 ml/min, and the measured energy expenditure did differ by -0.2% (+/- 14%) from mean predicted values using the Harris-Benedict equation. The VO2 was reduced by 15 +/- 2% with propofol, by 12 +/- 8% with midazolam, and by 10 +/- 5% with thiopentone. This was statistically significant compared to placebo treatment, as was the difference between propofol and thiopentone effects. All patients in these groups reached the defined sleep level, which was not achieved by the placebo and fentanyl groups. Placebo treatment changed the VO2 by 0.1% (+/- 2%). Fentanyl increased the VO2 by 5% (+/- 8%), which did not reach significance. In the fentanyl group the bolus application had to be stopped at a p(et)CO2 of 50 mm Hg in all patients. In the propofol, midazolam, and thiopentone groups the phasic changes of skin resistance were reduced to zero and the skin temperature increased from 27 +/- 2 degrees C to 32 +/- 2 degrees C. The fentanyl group showed an increase in changes of skin resistance without changes in temperature. CONCLUSIONS. Sleep induced by propofol, midazolam, or thiopentone to a clinically maximal desirable level in spontaneously breathing patients reduced VO2 by 10% to 15%. This level of sedation did not induce a relevant change in P(et)CO2 or SaO2. The effect of propofol appeared to be the most pronounced and least variable. This may be attributable to a more pronounced reduction in single-organ VO2 or to an undetected difference in level of sedation. Fentanyl did, in contrast to most publications on opioid effects, seem to increase VO2. Underlying mechanisms may be sought in an increased rate-pressure product and sympathetic activity on the basis of hypercapnia and changes in muscle tension.


Determination of membrane Peptide orientation by 1H-detected 2H NMR spectroscopy.
We demonstrate the application of the proton inverse detected deuteron (PRIDE) NMR technique to the measurement of the orientation of membrane-bound peptides with enhanced sensitivity. Gramicidin D, a transmembrane peptide, and ovispirin, a surface-bound peptide, were used as model systems. The peptides were 2H-labeled by 1H/2H exchange and oriented uniaxially on glass plates. The directly detected 2H spectra of both peptides showed only a strong D(2)O signal and no large quadrupolar splittings. In contrast, the PRIDE spectrum of gramicidin exhibited quadrupolar splittings as large as 281 kHz, consistent with its transmembrane orientation. Moreover, the large D(2)O signal in the directly detected 2H spectra was cleanly suppressed in the PRIDE spectrum. For ovispirin, the 1H indirectly detected 2H spectrum revealed a 104 kHz splitting and a zero-frequency peak. The former reflects the in-plane orientation of most of the helix axis, while the latter results from residues with a magic-angle orientation of the N-D bonds. These are consistent with previous 15N NMR results on ovispirin. The combination of PRIDE and exchange labeling provides an economical and sensitive method of studying membrane peptide orientations in lipid bilayers without the influence of D(2)O and with the ability to detect N-D bonds at the magic angle from the bilayer normal.


Synthetic High-Density Lipoprotein (sHDL) Inhibits Steroid Production in HAC15 Adrenal Cells.
High density lipoprotein (HDL) transported cholesterol represents one of the sources of substrate for adrenal steroid production. Synthetic HDL (sHDL) particles represent a new therapeutic option to reduce atherosclerotic plaque burden by increasing cholesterol efflux from macrophage cells. The effects of the sHDL particles on steroidogenic cells have not been explored. sHDL, specifically ETC-642, was studied in HAC15 adrenocortical cells. Cells were treated with sHDL, forskolin, 22R-hydroxycholesterol, or pregnenolone. Experiments included time and concentration response curves, followed by steroid assay. Quantitative real-time RT-PCR was used to study mRNA of 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase, lanosterol 14-α-methylase, cholesterol side-chain cleavage enzyme, and steroid acute regulatory protein. Cholesterol assay was performed using cell culture media and cell lipid extracts from a dose response experiment. sHDL significantly inhibited production of cortisol. Inhibition occurred in a concentration- and time-dependent manner and in a concentration range of 3μM-50μM. Forskolin (10μM) stimulated cortisol production was also inhibited. Incubation with 22R-hydroxycholesterol (10μM) and pregnenolone (10μM) increased cortisol production, which was unaffected by sHDL treatment. sHDL increased transcript levels for the rate-limiting cholesterol biosynthetic enzyme, 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase. Extracellular cholesterol assayed in culture media showed a positive correlation with increasing concentration of sHDL, whereas intracellular cholesterol decreased after treatment with sHDL. The current study suggests that sHDL inhibits HAC15 adrenal cell steroid production by efflux of cholesterol, leading to an overall decrease in steroid production and an adaptive rise in adrenal cholesterol biosynthesis.


Upper-limb assessment in people with Parkinson disease: is it a priority for therapists, and which assessment tools are used?
To investigate the frequency of physiotherapy and occupational therapy assessment of the upper limb (UL) in people with Parkinson disease (PD) and to identify the impairments and activity limitations assessed and the methods used. A custom-designed questionnaire was used to survey physiotherapists and occupational therapists with previous experience in managing people with PD, using targeted recruitment to invite physiotherapy conference attendees, clinicians employed in movement disorders programmes, and practitioners in neurology and gerontology to respond either on paper or online. Of the 190 respondents (122 physiotherapists, 68 occupational therapists), 54% reported consistently assessing the UL. A majority (>60%) assessed impairments specific to PD, but few quantified these using standardized measures. Activity limitations, largely relating to manual dexterity, were assessed using observational analysis (61%), non-standardized timed activities (46%), and standardized outcome measures (61%), most generic or developed for evaluating other neurological conditions. More than 10% of respondents could not identify an appropriate standardized measure. Slightly more than half of respondents regularly assessed the UL. Respondents reported widespread use of non-standardized methods to assess PD-specific impairments. Standardized measures were more frequently used to evaluate activity limitations, but despite the unique movement disorders associated with PD, the clinimetric properties of most of the tools identified have not been established in this population. Education and further clinimetric investigation of measures in use are needed to facilitate evidence-based practice in this area.


Pregnancy induces degenerative and regenerative changes in the autonomic innervation of the female reproductive tract.
The uterus is supplied with an extensive system of adrenergic nerves. The neurobiological properties of this innervation have been investigated in a series of studies primarily using the guinea-pig as model. The guinea-pig uterus is supplied from three different sources: the paracervical plexus, containing short adrenergic neurons; the inferior mesenteric ganglion; and a cranial source, probably the aorticorenal plexus, via nerves in the uterine suspensory ligaments. The nerve density is higher in the tubal end of the uterine horn and in the cervix than in the main part of the uterine horn. The turnover rate of transmitter is lower in the uterus than in a control organ, such as the heart. Noradrenaline levels in the uterus, but not the heart, are influenced by alterations in the endocrine milieu, e.g. during the oestrous cycle and after treatment with sex steroids. In the uterine tissue surrounding the conceptus during pregnancy, there is an early and drastic decay in various functional parameters related to the adrenergic nerve plexus and primarily reflecting a local, pregnancy-induced axonal degeneration. In the main part of the empty horn, in unilateral pregnancy, there is an extensive decay in various adrenergic functional parameters; these, however, reflect changes in a nerve plexus that has an essentially intact structure. No sign of functional impairment is seen in the adrenergic nerves of the uterine cervix. The increased turnover rate and reduced transmitter content in this region during late pregnancy may reflect increased frequency of firing of the adrenergic nerves. The tubal end of the uterine horn shows no signs of altered sympathetic function. This is the only part of the uterine horn that appears unaffected by pregnancy. The deficient recovery post partum of the changes in the uterine horn that previously contained the fetuses suggests permanent damage to the adrenergic nerve plexus after a pregnancy. The post partum recovery of changes seen in the previously empty horn, however, is more pronounced but still incomplete by comparison with the innervation before pregnancy. Studies on the adult human uterus indicate that similar events to those described for the guinea-pig model occur in human pregnancy.


Sex-specific associations of gestational glucose tolerance with childhood body composition.
To examine the associations of maternal gestational glucose tolerance with offspring body composition in late childhood. Among 958 women in the prebirth cohort Project Viva, glucose tolerance was assessed in the second trimester by nonfasting 50-g 1-h glucose challenge test (GCT), followed if abnormal by fasting 100-g 3-h oral glucose tolerance test (OGTT). We categorized women as normoglycemic (83.3%) if GCT was ≤140 mg/dL, isolated hyperglycemia (9.1%) if GCT was abnormal but OGTT normal, intermediate glucose intolerance (IGI) (3.3%) if there was one abnormal value on OGTT, or gestational diabetes mellitus (GDM) (4.5%) if there were two or more abnormal OGTT values. Using multivariable linear regression, we examined adjusted associations of glucose tolerance with offspring overall (N=958) and central (N=760) adiposity and body composition using dual X-ray absorptiometry (DXA) measured at the school-age visit (95±10 months). Compared with that in the male offspring of normoglycemic mothers, DXA fat mass was higher in male offspring of GDM mothers (1.89 kg [95% CI 0.33-3.45]) but not in male offspring of mothers with IGI (0.06 kg [-1.45 to 1.57]). DXA trunk-to-peripheral fat mass, a measure of central adiposity, was also somewhat higher in male offspring of GDM mothers (0.04 [-0.01 to 0.09]). In girls, DXA fat mass was higher in offspring of mothers with IGI (2.23 kg [0.12-4.34]) but not GDM (-1.25 kg [-3.13 to 0.63]). We showed no association of gestational glucose tolerance with DXA lean mass. In this study, only male offspring of GDM mothers manifested increased adiposity, whereas only female offspring of mothers with IGI did so. Sex differences in glycemic sensitivity may explain these findings.


Analysis of the Fungal Diversity and Community Structure in Sichuan Dark Tea During Pile-Fermentation.
The fungi present during pile-fermentation of Sichuan dark tea play a pivotal role in the development of its aroma and physical characteristics. Samples of tea leaves were collected on days 0 (YC-raw material), 8 (W1-first turn), 16 (W2-second turn), 24 (W3-third turn), and 32 (W4-out of pile) during pile-fermentation. High-throughput sequencing revealed seven phyla, 22 classes, 41 orders, 85 families, 128 genera, and 184 species of fungi. During fermentation, the fungal diversity index declined from the W1 to W3 stages and then increased exponentially at the W4 stage. A bar plot and heatmap revealed that Aspergillus, Thermomyces, Candida, Debaryomyces, Rasamsonia, Rhizomucor, and Thermoascus were abundant during piling, of which Aspergillus was the most abundant. Cluster analysis revealed that the W4 stage of fermentation is critical for fungal growth, diversity, and the community structure in Sichuan dark tea. This study revealed the role of fungi during pile-fermentation in the development of the essence and physical characteristics of Sichuan dark tea. This study comes under one of the Sustainable Development Goals of United Nations Organization (UNO) to "Establish Good Health and Well-Being."


Nanotube-Like Electronic States in [5,5]-C90 Fullertube Molecules.
Fullertubes, that is, fullerenes consisting of a carbon nanotube moiety capped by hemifullerene ends, are emerging carbon nanomaterials whose properties show both fullerene and carbon nanotube (CNT) traits. Albeit it may be expected that their electronic states show a certain resemblance to those of the extended nanotube, such a correlation has not yet been found or described. Here it shows a scanning tunneling microscopy (STM) and spectroscopy (STS) characterization of the adsorption, self-assembly, and electronic structure of 2D arrays of [5,5]-C90 fullertube molecules on two different noble metal surfaces, Ag(111) and Au(111). The results demonstrate that the shape of the molecular orbitals of the adsorbed fullertubes corresponds closely to those expected for isolated species on the grounds of density functional theory calculations. Moreover, a comparison between the electronic density profiles in the bands of the extended [5,5]-CNT and in the molecules reveals that some of the frontier orbitals of the fullertube molecules can be described as the result of the quantum confinement imposed by the hemifullerene caps to the delocalized band states in the extended CNT. The results thus provide a conceptual framework for the rational design of custom fullertube molecules and can potentially become a cornerstone in the understanding of these new carbon nanoforms.


Enhanced expression of tandem multimers of the antimicrobial peptide buforin II in Escherichia coli by the DEAD-box protein and trxB mutant.
The tandem multimeric expression of various peptides has been explored by many researchers. However, expression levels have usually not been proportional to the degree of multimerization. To increase the expression level in Escherichia coli of tandem multimers of a cationic antimicrobial peptide, buforin II, fused to an anionic peptide, we studied the effect of the DEAD-box protein and the trxB mutant on the expression of tandem multimers. An expression vector with a tac promoter was more effective in directing multimeric expression than one with a T7 promoter. The expression level of large multimers was substantially increased with the tac promoter, possibly through stabilization of long transcripts by synchronization of transcription and translation. Coexpression of the DEAD-box protein, an RNA-binding protein, with the T7 expression system increased the expression level of multimers, especially large multimers, due to protection of the long RNA transcripts. In addition, the use of the trxB mutant also enhanced the expression level of tandem multimers, which contain two cysteine residues at both ends of the monomeric unit. It seems that disulfide bonds formed in the multimers in the trxB mutant might help efficient charge neutralization for inclusion body formation of the multimers, resulting in enhancement of expression. Our results show that the expression of multimers can be improved through the stabilization of the long transcripts by the DEAD-box protein or the expression, under an oxidizing environment, of the trxB mutant in which covalent cross-links through disulfide bonds facilitate inclusion body formation of the multimeric fusion peptide.


Lower Face: Clinical Anatomy and Regional Approaches with Injectable Fillers.
The use of injectable fillers enables facial sculpting through treatment of volume depletion and modeling of facial contours. Injectable fillers are among the most frequently performed minimally invasive cosmetic procedures.However, treatment of the lower third of the face can be challenging and requires expertise in facial anatomy. In this article, the authors provide a comprehensive review of the anatomy of the lower third of the face, highlighting danger zones. In addition, the authors describe their preferred approach and detailed technique used in the treatment of each specific area, namely the jawline, prejowl sulcus, melomental folds, and lips.


Experimental study of the mechanism of the decrease in endocochlear d.c. potential after administration of nitrogen mustard-N-oxide.
20 mg/kg of body weight of nitrogen mustard-N-oxide hydrochloride ( NMNO ) causes a transient decrease in the magnitude of the endocochlear d.c. potential (EP) in the same manner as does nitrogen mustard (NM). NMNO does not, however, change the electrical resistance of the cochlear partition. When the administered dose of NMNO is increased to 40, 60 and 80 mg/kg body weight, a corresponding diminution in the EP is not observed. No ultrastructural changes in the stria vascularis are observed, even in animals administered 80 mg/kg body weight. The reduction of the EP with NMNO could not be attributed to a loss of the electrical insulator effect of the cochlear partition, or to a functional derangement of the stria vascularis. In the experimental animals whose organ of Corti was already destroyed with kanamycin sulfate (KM), NMNO did not reduce the magnitude of the EP. The organ of Corti may be central in understanding the mechanism of NMNO -induced reduction of the EP.


Transcriptional repressor REST drives lineage stage-specific chromatin compaction at Ptch1 and increases AKT activation in a mouse model of medulloblastoma.
In medulloblastomas (MBs), the expression and activity of RE1-silencing transcription factor (REST) is increased in tumors driven by the sonic hedgehog (SHH) pathway, specifically the SHH-α (children 3 to 16 years) and SHH-β (infants) subgroups. Neuronal maturation is greater in SHH-β than SHH-α tumors, but both correlate with poor overall patient survival. We studied the contribution of REST to MB using a transgenic mouse model (RESTTG ) wherein conditional NeuroD2-controlled REST transgene expression in lineage-committed Ptch1 +/- cerebellar granule neuron progenitors (CGNPs) accelerated tumorigenesis and increased penetrance and infiltrative disease. This model revealed a neuronal maturation context-specific antagonistic interplay between the transcriptional repressor REST and the activator GLI1 at Ptch1 Expression of Arrb1, which encodes β-arrestin1 (a GLI1 inhibitor), was substantially reduced in proliferating and, to a lesser extent, lineage-committed RESTTG cells compared with wild-type proliferating CGNPs. Lineage-committed RESTTG cells also had decreased GLI1 activity and increased histone H3K9 methylation at the Ptch1 locus, which correlated with premature silencing of Ptch1 These cells also had decreased expression of Pten, which encodes a negative regulator of the kinase AKT. Expression of PTCH1 and GLI1 were less, and ARRB1 was somewhat greater, in patient SHH-β than SHH-α MBs, whereas that of PTEN was similarly lower in both subtypes than in others. Inhibition of histone modifiers or AKT reduced proliferation and induced apoptosis, respectively, in cultured REST-high MB cells. Our findings linking REST to differentiation-specific chromatin remodeling, PTCH1 silencing, and AKT activation in MB tissues reveal potential subgroup-specific therapeutic targets for MB patients.


Detecting and Monitoring Porcine Hemagglutinating Encephalomyelitis Virus, an Underresearched Betacoronavirus.
Members of family Coronaviridae cause a variety of diseases in birds and mammals. Porcine hemagglutinating encephalomyelitis virus (PHEV), a lesser-researched coronavirus, can infect naive pigs of any age, but clinical disease is observed in pigs ≤4 weeks of age. No commercial PHEV vaccines are available, and neonatal protection from PHEV-associated disease is presumably dependent on lactogenic immunity. Although subclinical PHEV infections are thought to be common, PHEV ecology in commercial swine herds is unknown. To begin to address this gap in knowledge, a serum IgG antibody enzyme-linked immunosorbent assay (ELISA) based on the S1 protein was developed and evaluated on known-status samples and then used to estimate PHEV seroprevalence in U.S. sow herds. Assessment of the diagnostic performance of the PHEV S1 ELISA using serum samples (n = 924) collected from 7-week-old pigs (n = 84; 12 pigs per group) inoculated with PHEV, porcine epidemic diarrhea virus, transmissible gastroenteritis virus, porcine respiratory coronavirus, or porcine deltacoronavirus showed that a sample-to-positive cutoff value of ≥0.6 was both sensitive and specific, i.e., all PHEV-inoculated pigs were seropositive from days postinoculation 10 to 42, and no cross-reactivity was observed in samples from other groups. The PHEV S1 ELISA was then used to estimate PHEV seroprevalence in U.S. sow herds (19 states) using 2,756 serum samples from breeding females (>28 weeks old) on commercial farms (n = 104) with no history of PHEV-associated disease. The overall seroprevalence was 53.35% (confidence interval [CI], ±1.86%) and herd seroprevalence was 96.15% (CI, ±3.70%).IMPORTANCE There is a paucity of information concerning the ecology of porcine hemagglutinating encephalomyelitis virus (PHEV) in commercial swine herds. This study provided evidence that PHEV infection is endemic and highly prevalent in U.S. swine herds. These results raised questions for future studies regarding the impact of endemic PHEV on swine health and the mechanisms by which this virus circulates in endemically infected populations. Regardless, the availability of the validated PHEV S1 enzyme-linked immunosorbent assay (ELISA) provides the means for swine producers to detect and monitor PHEV infections, confirm prior exposure to the virus, and to evaluate the immune status of breeding herds.


The outcomes of salvage surgery for vascular injury in the extremities: a special consideration for delayed revascularization.
A seven years retrospective study was performed in 45 consecutive vascular injuries in the extremities to investigate the pattern of injuries, managements and outcomes. Motor-vehicle accidents were the leading cause of injuries (80%), followed by industrial injuries (11.1%) and iatrogenic injuries (4.4%). Popliteal and brachial artery injuries were commonly involved (20%). Fifteen (33.3%) patients had fractures, dislocation or fracture dislocation around the knee joint and 6 (13.3%) patients had soft tissue injuries without fracture. Traumatic arterial transection accounted for 34 (75.6%) cases, followed by laceration in 7 (15.6%) and 9 (6.7%) contusions. Associated nerve injuries were seen in 8 (17.8 %) patients using intra-operative findings as the gold standard, both conventional angiogram (CA) and computerized tomography angiogram (CTA) had 100% specificity and 100% sensitivity in determining the site of arterial injuries. The mean ischemic time was 25.31 hours (4 - 278 hours). Thirty-three (73.3 %) patients were treated more than 6 hours after injury and 6 patients underwent revascularization after 24 hours; all had good collateral circulation without distal pulses or evidence of ischemic neurological deficit. The mean ischemic time in 39 patients who underwent revascularization within 24 hours was 13.2 hours. Delayed amputation was performed in 5 patients (11.1%). Of the 6 patients who underwent delayed revascularization, one patient had early amputation, one -had delayed amputation following infection and multiple flap procedures while the rest of the patients' limbs survived. Joint stiffness was noted in 10 patients (22.2%) involving the knee joint, elbow and shoulder in two patients each. Infection was also noted in 5 patients (11.1%) with two of them were due to infected implants. Other complications encountered included nonunion (2 patients, 4.4%), delayed union (1 patient, 2.2%),limb length discrepancy (1 patient, 2.2%), hematoma (1 patient, 2.2%) and leaking anastomosis in one patient (2.2%). Volkmann's ischemic contracture occurred in 3 (6.7%) patients. There was no complication noted in 8 (17.8%) patients Three patients (6.7%) died of whom two were not due to vascular causes. We conclude that early detection and revascularization of traumatic vascular injuries is important but delayed revascularization also produced acceptable results.


Free luteinizing-hormone beta-subunit in normal subjects and patients with pituitary adenomas.
Most clinically nonfunctioning pituitary adenomas (NFPA) are found to be gonadotropinomas when assessed by immunocytochemistry. However, they are rarely associated with increased basal plasma levels of FSH, LH and/or alpha-subunit. It has been claimed that the paradoxical free LHbeta response to TRH may be a useful clinical tool for determining the gonadotropic nature of NFPA. We used a very specific and sensitive immunoradiometric assay (IRMA) for free LHbeta measurement and another specific IRMA to check the absence of free CGbeta, to study normal subjects and 26 patients with NFPA. Basal plasma levels of LHbeta were undetectable in normal men and premenopausal women in the early follicular phase. In contrast, normal postmenopausal women had increased basal plasma LHbeta, parallel to dimeric LH and alpha-subunit levels. In healthy subjects, stimulation with GnRH elicited an increase in LHbeta while TRH was ineffective. In patients with NFPA, LHbeta hypersecretion was found basally and/or after stimulation with TRH in 3 of 16 men, 3 of 5 premenopausal women, and 1 of 5 postmenopausal women, i.e. 7 of 26 patients (26%). In 3 of these 7 cases, alpha-subunit and/or FSH levels were also increased. The LHbeta measurement was thus truly informative on the gonadotropic nature of NFPA in only 4 out of 26 cases (15%). In addition, increased LHbeta levels and/or a positive response of free LHbeta to TRH was observed in 3 patients with pure prolactinomas but in no patients with GH-secreting adenomas. Thus, using this very sensitive and specific IRMA, free LHbeta measurement is rarely helpful for determining the gonadotropic nature of NFPA.


alpha-Glucosidase inhibitors have a prolonged antiviral effect against hepatitis B virus through the sustained inhibition of the large and middle envelope glycoproteins.
Previous work has shown that the secretion of enveloped hepatitis B virus (HBV) DNA and the HBV middle envelope protein (MHBs) are sensitive to glucosidase inhibition. Here, it is shown that HBV DNA secretion remains depressed after the removal of the glucosidase inhibitor and long after glucosidase function returns to normal. For example, glyco-processing and the secretion of alpha-1 anti-trypsin returned to normal within 3 h of the removal of the glucosidase inhibitor. In contrast, the secretion of HBV did not return to normal for more than 7 days after the removal of the inhibitor. Consistent with the inhibition of HBV virion secretion, the levels of HBV L and HBV M proteins were also reduced by treatment with the glucosidase inhibitor and remained reduced for 7 days after compound withdrawal. The implications of the prolonged antiviral effect against HBV and the use of glucosidase inhibitors as antiviral agents are discussed.


[Experimental study on characteristics of biodiesel exhausted particle].
A particle emission experiment of a direct-injection turbocharged diesel engine with biodiesel and diesel was carried out. A pump of 80 L/min and fiber glass filters with diameter of 90 mm was used to sample engine particles in exhaust pipe. The size distribution, soluble organic fraction (SOF) and 16 polycyclic aromatic hydrocarbons (PAHs) of particles were analyzed by a laser diffraction particle size analyzer and GC-MS. The results indicate that the volume weighted size distribution of biodiesel particle is single-peak and its median diameter d(0.5) and mean diameter d32 are decreased with the increasing speed. At the high speed the d32 and d(0.5) of biodiesel are larger than those of diesel, and quite the contrary at the low speed. SOF mass concentration and mass percentage of biodiesel are 12.3 - 31.5 mg/m3 and 38.2% - 58.0% respectively, which are much higher than those of diesel. The total PAHs emission concentration of biodiesel is 2.9 - 4.7 microg/m3 lower than that of diesel as much as 29.1% - 92.4%.


Short communication: limit feeding affects behavior patterns and feeding motivation of dairy heifers.
The study objective was to assess the effects of limit feeding dairy heifers on behavior patterns and feeding motivation. Ten Holstein heifers (291.6±39.2d of age, weighing 324.2±61.2kg; mean ± SD) were exposed to each of 2 dietary treatments, in a random order, over 2 successive 26-d treatment periods (14-d adaptation period and a 12-d data collection period) using a crossover design: (1) a high-forage total mixed ration (TMR), provided ad libitum (CON) and (2) a low-forage TMR, limit-fed at 2.05% body weight (LF). Heifers were fed daily at 1100h and motivation to access a low-nutritive feedstuff (straw) was assessed using a push-door apparatus at 2 time points: 3h after feed delivery (1400h) and 21h after feed delivery (0800h). The amount of weight pushed, weight pushed as percentage of body weight, and latency to access the push door were recorded on 3 different days for each heifer at each time point on each treatment. When fed CON, heifers had greater dry matter intake (12.9 vs. 7.2kg/d), greater feeding time (209.3 vs. 82.4min/d), greater ruminating time (452.2 vs. 318.3min/d), and slower rates of intake (0.06 vs. 0.09kg of dry matter/min) than when fed LF. Heifers fed LF pushed more weight as a percentage of body weight at 3h (4.5 vs. 1.9%) and 21h (9.3 vs. 2.8%) after feed delivery. At both 3 and 21h after feed delivery, latency to access the door was shorter for the LF heifers compared with the CON heifers (65 vs. 145 s). These results indicate that, in addition to decreasing feeding time, limit feeding increases motivation of heifers to access a low-nutritive feedstuff, possibly due to lack of satiety resulting from lack of physical fill or insufficient time spent foraging.


Serum immunoglobulin levels in hepatitis-B antigen-positive and hepatitis-B antigen-negative acute hepatitis.
IgG, IgA and IgM levels were measured in the serum of 73 patients with hepatitis-B antigen-positive (virus-B) acute hepatitis and 48 patients with hepatitis-B antigen-negative (virus-A) hepatitis. The mean serum IgM level on admission was significantly higher in virus-A than in virus-B hepatitis (p less than 0,01), and remained so for 2-3 weeks. However, the overlap between the individual figures in the two forms of the disease was so great as to make the estimation of little diagnostic value. The mean serum IgG and IgA levels at the time of admission were not significantly different in the patients with and without hepatitis-B anti-genaemia.


Biceps tendon tenodesis for posterolateral instability of the knee. An in vitro study.
The effects of biceps tendon tenodesis on internal-external and varus-valgus laxity were measured using fresh-frozen cadaveric specimens that had undergone sequential sectioning of the posterolateral structures and of the fibular collateral ligament. Tenodesis (using 89 N graft tension and a fixation point located 1 cm anterior to the fibular collateral ligament's insertion on the femur) was effective in restoring external rotation and varus laxity; the procedure actually overconstrained external tibial rotation at all flexion positions and varus angulation at 60 degrees and 90 degrees of flexion. Internal rotation and valgus laxity were unaffected by the tenodesis procedure. The anterior fixation point was more effective in reducing laxity than a fixation point located 1 cm proximal to the fibular collateral ligament insertion. Tenodesis using the proximal fixation point, which was nonisometric, did not restore external rotation and varus laxities to intact values at 60 degrees and 90 degrees of knee flexion. Graft tension (45 or 89 N) had no measurable effect on the results of the tenodesis. This study has demonstrated that the biceps tenodesis procedure is effective for reducing static laxity in the knee with posterolateral instability.


IL-4R{alpha}-responsive smooth muscle cells increase intestinal hypercontractility and contribute to resistance during acute Schistosomiasis.
Interleukin-(IL)-4 and IL-13 signal through heterodimeric receptors containing a common IL-4 receptor-alpha (IL-4Ralpha) subunit, which is important for protection against helminth infections, including schistosomiasis. Previous studies demonstrated important roles for IL-4Ralpha-responsive hematopoietic cells, including T cells and macrophages in schistosomiasis. In this study, we examined the role of IL-4Ralpha responsiveness by nonhematopoietic smooth muscle cells during experimental acute murine schistosomiasis. Comparative Schistosoma mansoni infection studies with smooth muscle cell-specific IL-4Ralpha-deficient (SM-MHC(cre)IL-4Ralpha(-/flox)) mice, heterozygous control (IL-4Ralpha(-/flox)) mice, and global IL-4Ralpha-deficient (IL-4Ralpha(-/-)) mice were conducted. S. mansoni-infected SM-MHC(cre)IL-4Ralpha(-/flox) mice showed increased weight loss and earlier mortalities compared with IL-4Ralpha(-/flox) mice, despite comparable T(H)2/type 2 immune responses. In contrast to highly susceptible IL-4Ralpha-deficient mice, increased susceptibility in SM-MHC(cre)IL-4Ralpha(-/flox) mice was not accompanied by intestinal tissue damage and subsequent sepsis. However, both susceptible mutant mouse strains failed to efficiently expel eggs, demonstrated by egg reduction in the feces compared with control mice. Reduced egg expulsion was accompanied by impaired IL-4/IL-13-mediated hypercontractile intestinal responses, which was present in the more resistant control mice. Together, we conclude that IL-4Ralpha responsiveness by smooth muscle cells and subsequent IL-4- and IL-13-mediated hypercontractility are required for host protection during acute schistosomiasis to efficiently expel S. mansoni eggs and to prevent premature mortality.


Higenamine Reduces Fine-Dust-Induced Matrix Metalloproteinase (MMP)-1 in Human Keratinocytes.
Environmental pollutants such as fine dust are increasingly linked to premature skin aging. In this study, we investigated the protective effects of higenamine, a natural plant alkaloid, against fine-dust-induced skin aging in human keratinocytes (HaCaT cells). We found that higenamine significantly attenuated fine-dust-induced expression of matrix metalloproteinase-1 (MMP-1), a key enzyme involved in collagen degradation. Furthermore, higenamine was found to modulate fine-dust-induced AP-1 and NF-κB transactivation, which are crucial factors for MMP-1 transcription. Higenamine also impeded fine-dust-induced phosphorylation in specific pathways related to AP-1 and NF-κB activation, and effectively alleviated reactive oxygen species (ROS) production, a key factor in oxidative stress caused by fine dust exposure. These results suggest that higenamine exerts protective effects against fine-dust-induced skin aging, primarily through its MMP-1 inhibitory properties and ability to mitigate ROS-induced oxidative damage. Our data highlight the potential of higenamine as an effective ingredient in skincare products designed to combat environmental skin damage.


The Pseudomonas aeruginosa pfpI gene plays an antimutator role and provides general stress protection.
Hypermutator Pseudomonas aeruginosa strains, characterized by an increased spontaneous-mutation rate, are found at high frequencies in chronic lung infections. Hypermutability is associated with the loss of antimutator genes related to DNA repair or damage avoidance systems. Only a few antimutator genes have been described in P. aeruginosa, although there is some evidence that additional genes may be involved in naturally occurring hypermutability. In order to find new P. aeruginosa antimutator genes, we constructed and screened a library of random insertions in the PA14 strain. Some previously described P. aeruginosa and/or Escherichia coli antimutator genes, such as mutS, mutL, uvrD, mutT, ung, and mutY, were detected, indicating a good coverage of our insertional library. One additional mutant contained an insertion in the P. aeruginosa PA14-04650 (pfpI) gene, putatively encoding a member of the DJ-1/ThiJ/PfpI superfamily, which includes chaperones, peptidases, and the Parkinson's disease protein DJ-1a. The pfpI-defective mutants in both PAO1 and PA14 showed higher spontaneous mutation rates than the wild-type strains, suggesting that PfpI plays a key role in DNA protection under nonstress conditions. Moreover, the inactivation of pfpI resulted in a dramatic increase in the H(2)O(2)-induced mutant frequency. Global transcription studies showed the induction of bacteriophage Pf1 genes and the repression of genes related to iron metabolism, suggesting that the increased spontaneous-mutant frequency may be due to reduced protection against the basal level of reactive oxygen species. Finally, pfpI mutants are more sensitive to different types of stress and are affected in biofilm formation.


[Influence of gastric juice on human amylase activity].
Although it is well established that amylase activity is inactivated by acid, little is known about the influence of native gastric juice on amylase activity. We investigated the effect of gastric juice, acid, and pepsin on both pancreatic (P-) and salivary (S-) isoamylase activities in vitro. S- and P-isoamylase activities were completely inactivated by HCl (10 mEq/l), but 20% of each isoamylase activity remained when bovine serum albumin (BSA) (3%) was added. However pepsin (0.2 tyrosine mg/ml/min) inactivated both isoamylase activities even though BSA was added. When saliva or pancreatic juice was mixed with gastric juice, both isoamylase activities were detected until the mixing ratio was 3:7 (gastric juice:saliva or pancreatic juice, v/v). It is suggested that some of both isoamylase activities could survive after exposure to gastric juice in the stomach and duodenum, depending on the concentrations of acid and protein in gastric juice, and mixing ratio to gastric juice.


ABCB1 C3435T polymorphism influences methotrexate sensitivity in rheumatoid arthritis patients.
Methotrexate (MTX) is most widely used for the treatment of rheumatoid arthritis (RA). However, it has certain drawbacks with regard to individual differences in its therapeutic effects as well as the differences in the patients' response to MTX therapy. We investigated whether multi-drug resistance-1 (ABCB1) C3435T, reduced folate carrier-1 (RFC1) G80A, 5-aminoimidazole-4-carboxamide ribonucleotide transformylase (ATIC) C347G and a 6bp-deletion polymorphism in the 3'-untranslated region of the thymidylase synthase (TYMS) gene are predictive of MTX sensitivity and its adverse effects. Patients whose last maintenance dosage of MTX was <or= 6 mg/week were regarded as responders, while patients whose last maintenance dosage of MTX was > 6 mg/week or those in whom MTX therapy was changed due to poor response to MTX were regarded as non-responders. The data of 124 RA patients who had received MTX treatment were retrospectively analyzed for polymorphisms in the ABCB1, RFC1, ATIC and TYMS genes, MTX sensitivity and MTX toxicity. There were no significant differences in MTX sensitivity among the genotypes of RFC1, ATIC and TYMS genes. ABCB1 3435TT cases included statistically significantly more non-responders than 3435CC cases according to univariate analysis (crude odds ratio (OR) = 8.91, p = 0.001) and multivariate analysis (adjusted OR = 8.78, p = 0.038). There were no significant differences in MTX toxicity among the genotypes of all the genes. These results suggested that the genetic diagnosis of ABCB1 C3435T can be applied to determine MTX sensitivity for the treatment of RA patients. However, further pharmacokinetics studies are required in this regard.


[Protective effects of Guangdong Liangcha grandes on restraint stress-induced liver damage in mice].
To investigate the effect of Guangdong Liangcha Keli on restraint stress-induced liver damage in mice. Thirty-five male C57BL/6J mice of 7 weeks old were divided into 5 groups randomly with 7 mice in each group: normal group, restraint stress group, 250 mg kg(-1) Vitamin C, Guangdong Liangcha Keli 500 mg kg(-1) and 250 mg kg(-1). After 18 hr restraint stress, the ALT acitivity in plasma, MDA level in plasma and liver, GSH content, GSH-PX and GST activities, NO level and ORAC value in liver were determined. Compared with restraint model group, Guangdong Liangcha Keli could markedly reduce ALT activity (92.75 +/- 1.91 vs 39.29 +/- 2.56, 32.69 +/- 1.46) U L(-1), and protect the liver damage induced by oxidative stress. In addition, Guangdong Liangcha Keli could effectively increase the ORAC value, GSH content, GSH-PX activity and GST activity and reduce the MDA level and NO level in liver. Oral treatment of Guangdong Liangcha Keli is found to reduce restraint stress-induced liver damage in terms of above mentioned biochemical parameters, and these protective effects may be related to its free radical scavenging activity and lipid peroxidation inhibitory effects.


BRAT1-associated neurodegeneration: Intra-familial phenotypic differences in siblings.
Recessive mutations in BRAT1 cause lethal neonatal rigidity and multifocal seizure syndrome, a phenotype characterized by neonatal microcephaly, hypertonia, and refractory epilepsy with premature death by age 2 years. Recently, attenuated disease variants have been described, suggesting that a wider clinical spectrum of BRAT1-associated neurodegeneration exists than was previously thought. Here, we report two affected siblings with compound heterozygous truncating mutations in BRAT1 and intra-familial phenotypic heterogeneity, with a less severe disease course in the female sibling. This phenotypic variability should be taken into account when treating patients with BRAT1-associated neurodegenerative disease. Mildly affected individuals with BRAT1 mutations show that BRAT1 must be considered as a cause in childhood refractory epilepsy and microcephaly with survival beyond infancy. © 2016 Wiley Periodicals, Inc.


Extraskeletal Ewing's Sarcoma Masquerading as Infantile Benign Neck Mass.
Despite being the second most common malignant bone tumor, Ewing's sarcoma remains uncommon in younger children and seldom seen in neonates and infants. Extraskeletal locations are even rarer, hardly ever suspected, and often misdiagnosed, causing delays in management. The histologic similarities of Ewing's sarcoma to more common pediatric small-blue-round-cell tumors such as lymphoma and neuroblastoma necessitate immunohistochemistry and molecular genetics for clinching the diagnosis. We report a soft-tissue Ewing's sarcoma in a 4-month-old female infant masquerading as a benign neck mass clinically, radiologically, cytologically, and intraoperatively. We also reviewed literature for any existing guidelines on when to biopsy neck masses in the pediatric population.


Antimicrobial resistance of Campylobacter jejuni and Campylobacter coli with special reference to plasmid profiles of Japanese clinical isolates.
A total of 111 clinical isolates of Campylobacter jejuni and 10 clinical isolates of Campylobacter coli were characterized by their susceptibility to nine antimicrobial agents and by their plasmid profiles on agarose gel electrophoresis. All of the C. jejuni isolates were susceptible to chloramphenicol, ciprofloxacin, erythromycin, kanamycin, and nalidixic acid, but 55% were tetracycline resistant. In the 10 C. coli isolates, a high prevalence of multiple-antibiotic resistance was noted. Plasmids were found in 82% of the tetracycline-resistant and 15% of the tetracycline-susceptible C. jejuni isolates. Tetracycline resistance in six randomly selected C. jejuni isolates, which contained 50- or 135-kilobase (kb) plasmids, was transferred by conjugation to a Campylobacter fetus subsp. fetus recipient with recovery of a 50- or a 45-kb plasmid from transconjugants. From one multiple-antibiotic-resistant C. coli isolate, resistance to tetracycline, kanamycin, and chloramphenicol was transferred concomitantly with a 58-kb plasmid, pNR9589. Nonconjugative 98-kb plasmids, pNR9131 and pNR9581, from C. coli isolates with resistance to tetracycline, kanamycin, and erythromycin were shown by cloning experiments to code for at least kanamycin resistance. Restriction digests revealed that 50-kb plasmids from tetracycline-resistant C. jejuni isolates were identical, although plasmids from multiple-antibiotic-resistant C. coli isolates shared partial DNA homology to each other. Cloning of the kanamycin and chloramphenicol resistance genes of pNR9589 into Escherichia coli showed that the two genes are closely linked or clustered. Double-digestion analysis of the fragments encoding the kanamycin resistance of pNR9131, pNR9581, and pNR9589 showed that these three plasmids contain a common fragment related to kanamycin resistance.


The Path From Heart Failure to Cardiac Transplant.
Heart failure is a progressive and fatal disease impacting millions of American each year. Divided into stages, heart failure presents with progressive symptoms requiring a wide range of medical treatments. Treatments include diet and lifestyle changes, medications, electrical therapies (defibrillator and/or cardiac resynchronization therapy), as well as mechanical circulatory support. Cardiac transplant is the gold standard treatment of heart failure, although the availability of donors limits the utility of a cardiac transplant. This article outlines heart failure treatments and the indications, contraindications, and pretransplant evaluation for a cardiac transplant. Information on the allocation of donor hearts and donor characteristics is also included for the reader.


A COMPARISON OF TWO STICKLEBACKS.
We present results of an experiment designed to address fundamental issues in the ecology and evolution of plastic trophic morphology: (1) Is observed plasticity adaptive? (2) How much interspecific morphological variation is the result of plasticity? (3) Have different selective regimes resulted in the evolution of different degrees of plasticity? (4) Is genetic variation for phenotypic plasticity present in contemporary populations? We raised fish from two recently diverged species of freshwater threespine sticklebacks on two different diets representative of the natural prey of the two species. Both species exhibited morphological plasticity in an adaptive direction: each species more closely resembled the other when raised on the latter's diet. Dietreversal reduced the natural morphological gap between these two species, -1% to 58%, depending on the trait. One species is known to have a more variable diet in the wild than the other species, and we found that it also exhibited the greater amount of morphological plasticity. Given that the two species have recently diverged, this result is compelling evidence that diet variability is important in the evolution of plastic trophic morphology. Finally, by using a full-sib experimental design, we demonstrated that genetic variation for morphological plasticity exists in contemporary populations, thus confirming that plasticity has evolutionary potential.


Particle Morphology Analysis of Biomass Material Based on Improved Image Processing Method.
Particle morphology, including size and shape, is an important factor that significantly influences the physical and chemical properties of biomass material. Based on image processing technology, a method was developed to process sample images, measure particle dimensions, and analyse the particle size and shape distributions of knife-milled wheat straw, which had been preclassified into five nominal size groups using mechanical sieving approach. Considering the great variation of particle size from micrometer to millimeter, the powders greater than 250 μm were photographed by a flatbed scanner without zoom function, and the others were photographed using a scanning electron microscopy (SEM) with high-image resolution. Actual imaging tests confirmed the excellent effect of backscattered electron (BSE) imaging mode of SEM. Particle aggregation is an important factor that affects the recognition accuracy of the image processing method. In sample preparation, the singulated arrangement and ultrasonic dispersion methods were used to separate powders into particles that were larger and smaller than the nominal size of 250 μm. In addition, an image segmentation algorithm based on particle geometrical information was proposed to recognise the finer clustered powders. Experimental results demonstrated that the improved image processing method was suitable to analyse the particle size and shape distributions of ground biomass materials and solve the size inconsistencies in sieving analysis.


The 48-week efficacy of once-daily saquinavir/ritonavir in patients with undetectable viral load after 3 years of antiretroviral therapy.
To evaluate the efficacy and safety of once-daily saquinavir-soft-gel-capsules/ritonavir (SQV-SGC/RTV) 1600 mg/100 mg plus dual nucleoside reverse transcriptase inhibitors (NRTIs) in HIV-infected patients with plasma viral load (pVL) <50 HIV-1 RNA copies/mL following 3 years of antiretroviral therapy. A total of 69 patients with pVL <50 copies/mL after 162 weeks of antiretroviral treatment started SQV-SGC/RTV 1600 mg/100 mg once-daily while continuing dual NRTIs. Previous treatment consisted of 66 weeks of treatment with a half/full dose of zidovudine (ZDV)/zalcitabine (ddC), followed by 2 years of SQV-SGC twice a day (bid) plus ZDV/lamivudine (3TC) or didanosine (ddI)/stavudine (d4T). Efficacy (pVL), safety and immunological changes (CD4 cell counts) were evaluated after 48 weeks in this open-label, single-arm prospective study. SQV-SGC/RTV once-daily was well tolerated. No patient changed regimens or was lost to follow-up. After 48 weeks, 63 of 69 patients (91%) had pVL <50 copies/mL (five of the six remaining patients had pVL <400 copies/mL, and one patient had an unexplained rise to 39 500 copies/mL, which decreased to <50 copies/mL 12 weeks later). Median CD4 count increased from 534 cells/muL at the start of the SQV-SGC/RTV once-daily treatment to 664 cells/muL (P<0.001). Compared to the preceding 48 weeks on bid SQV-SGC, the CD4 cell count improved significantly on once-daily SQV-SGC/RTV (P<0.001). These data support the use of SQV-SGC/RTV 1600 mg/100 mg once-daily with two NRTIs as a convenient, safe and cost-saving regimen to maintain viral suppression and CD4 counts for 48 weeks in this preselected cohort on highly active antiretroviral therapy (HAART) with pVL <50 copies/mL. The CD4 count rise may be a result of continued immune reconstitution in patients with well-controlled infection.


ApoE4 expression disrupts tau uptake, trafficking, and clearance in astrocytes.
Tauopathies are a collection of neurodegenerative diseases characterized by the accumulation of pathogenic aggregates of the microtubule-associated protein tau. Despite the prevalence and diversity of tau astrogliopathy in tauopathies, the interactions between astrocytes and tau in the brain, and the influence of neurodegenerative genetic risk factors like the apolipoprotein E4 (apoE4) isoform, are largely unknown. Here, we leveraged primary and immortalized astrocytes expressing humanized apoE isoforms to characterize the mechanisms by which astrocytes interact with and eliminate extracellular tau, and the influence of apoE genotype on these processes. Our work indicates that astrocytes rapidly internalize, process, and release tau via an exosomal secretory mechanism under physiological conditions. However, we found that apoE4 disrupted these processes in comparison to apoE3, resulting in an astrocytic phenotype prone to intracellular tau accumulation. Furthermore, exposure to repetitive mild traumatic brain injuries exacerbated the apoE4-induced impairments in tau processing and elimination by astrocytes in apoE4 targeted-replacement mice. The diminished ability of apoE4 astrocytes to eliminate extracellular tau can lead to an accumulation of pathogenic tau, which induces mitochondrial dysfunction, as demonstrated by our studies. In total, our findings suggest that the apoE4 isoform lowers the threshold of astrocytic resilience to pathogenic tau, rendering them susceptible to bioenergetic deficits in the early stages of neurodegenerative diseases such as traumatic brain injury, potentially contributing to neurological decline.


[Effectiveness of specific immunotherapy in the treatment of children and youngsters suffering from atopic dermatitis. Part III. Serum concentrations of selected immunologic parameters].
Etiology and pathomechanism of atopic dermatitis still remains partially unclear and therefore contemporary methods of treatment are not always satisfactory from the clinical standpoint. The aim of this study was to evaluate selected immunological parameters (tIgE, ECP, sIL-2R, IFN-gamma, IL-4,IL-5) in sera of atopic dermatitis patients in the course of specific immunotherapy performed for the time period of 3 years. Novo-Helisen Depot allergy vaccines of appropriate composition were used for the treatment of 36 children and youngsters with atopic dermatitis, allergic to house dust mites (24 patients) and grass pollen allergens (12 patients). The control group consisted of 20 patients with atopic dermatitis and analogous IgE-mediated airborne allergy who were treated with conventional methods. There was a clear difference between two investigated groups of patients in terms of immunological parameters. In the group treated with allergy vaccines serum concentrations of total IgE and ECP tended to decrease (p < 0.001) as well as sIL-2R (p < 0.01). On the contrary in the control group serum tIgE increased and IL-4 as well as IL-5 concentrations tended to increase significantly (p < 0.01; p < 0.05 respectively).


Concurrent sodium channelopathies and amyotrophic lateral sclerosis supports shared pathogenesis.
Amyotrophic lateral sclerosis (ALS) is an invariably fatal adult-onset neurodegenerative disorder; approximately 10% of ALS is monogenic but all ALS exhibits significant heritability. The skeletal muscle sodium channelopathies are a group of inherited, non-dystrophic ion channel disorders caused by heterozygous point mutations in the SCN4A gene, leading to clinical manifestations of congenital myotonia, paramyotonia, and periodic paralysis syndromes. We provide clinical and genetic evidence of concurrence of these two rare disorders which implies a possible shared underlying pathophysiology in two patients. We then identify an enrichment of ALS-associated mutations in another sodium channel, SCN7A, from whole genome sequencing data of 4495 ALS patients and 1925 controls passing multiple testing correction (67 variants, p = 0.0002, Firth logistic regression). These findings suggest dysfunctional sodium channels may play a role upstream in the pathogenesis of ALS in a subset of patients, potentially opening the door to novel personalized medicine approaches.


Vibrio parahaemolyticus becomes lethal to post-larvae shrimp via acquiring novel virulence factors.
Translucent post-larvae disease (TPD), caused by Vibrio parahaemolyticus (Vp TPD), has become an emerging shrimp disease, affecting more than 70%-80% of coastal shrimp nurseries in China in spring 2020. Here, we investigated the key virulence factors of Vp TPD by analyzing protein fragments, related genomic information, as well as experimental challenge tests. After investigating the toxic effects of purified protein fragments with different molecular weights (MWs) from Vp TPD, we found that only the protein fragments with MW >100 kDa showed similar lethality to live Vp TPD in the experimental challenge test using post-larvae shrimp. Meanwhile, similar histopathological changes exhibiting in the hepatopancreas and midgut of the diseased individuals were observed in the post-larvae shrimp challenged with either bacterial protein fragments (MW >100 kDa) or live Vp TPD. Based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and mass spectrometry analyses, two novel proteins, Vibrio high virulent protein (VHVP)-1 and VHVP-2, were identified as the candidates of key virulence factors to cause TPD. Moreover, VHVP-1 and VHVP-2 were found to be encoded by two genes (vhvp-1 and vhvp-2) tandemly located on a 187,791-bp plasmid and were predicted to depend on the same promoter following a comparative genomic analysis. Further epidemiological investigation and challenge test indicated that the V. parahaemolyticus isolate carrying only the vhvp-1 gene and lacking vhvp-2 gene could not cause mortality of experimental Penaeus vannamei post-larvae. The mutant (Δvhvp-2) by deleting vhvp-2 gene could only cause 4.92% of accumulative mortality of post-larvae that is similar to the non-Vp TPD Vibrio strain. Additionally, the complemented strains, Δvhvp-2/pBT3-vhvp-2 and Δvhvp-2/pwtCas9-vhvp-2, showed similar virulence to the wild-type Vp TPD. The results demonstrated that V. parahaemolyticus becomes lethal to post-larval shrimp by acquiring the VHVP-2 virulence factor. This study sheds light on further investigations of the pathogenic mechanism of Vp TPD and the development of strategies for early diagnosis of TPD in shrimp hatcheries. As a severe emerging shrimp disease, TPD has heavily impacted the shrimp aquaculture industry and resulted in serious economic losses in China since spring 2020. This study aimed to identify the key virulent factors and related genes of the Vp TPD, for a better understanding of its pathogenicity of the novel highly lethal infectious pathogen, as well as its molecular epidemiological characteristics in China. The present study revealed that a novel protein, Vibrio high virulent protein-2 (MW >100 kDa), is responsible to the lethal virulence of V. parahaemolyticus to shrimp post-larvae. The results are essential for effectively diagnosing and monitoring novel pathogenic bacteria, like Vp TPD, in aquaculture shrimps and would be beneficial to the fisheries department in early warning of Vp TPD emergence and developing prevention strategies to reduce economic losses due to severe outbreaks of TPD. Elucidation of the key virulence genes and genomics of Vp TPD could also provide valuable information on the evolution and ecology of this emerging pathogen in aquaculture environments.


Molecular analysis of chromosome 20q deletions associated with myeloproliferative disorders and myelodysplastic syndromes.
Acquired deletions of the long arm of chromosome 20 are found in several hematologic conditions and particularly in the myeloproliferative disorders and myelodysplastic syndromes. The spectrum of diseases associated with 20q deletions suggests that such deletions may mark the site of a tumor suppressor gene that contributes to the regulation of normal multipotent hematopoietic progenitors. We present here the first detailed molecular analysis of 20q deletions associated with myeloid disorders. Thirty-four microsatellite primer pairs corresponding to loci on 20q have been used to study DNA samples from two cell lines and from highly purified peripheral blood granulocytes obtained from seven patients. In addition, Southern analysis of cell line DNA has been performed using 19 DNA probes that map to 20q. Three conclusions can be drawn from our results. Firstly, molecular heterogeneity of both centromeric and telomeric breakpoints was demonstrated, thus supporting the existence of a tumor suppressor gene on 20q. In addition many of the breakpoints have been mapped to small genetic intervals. Secondly, our results define a commonly deleted region of 16-21 cM which contains ADA, PLC1, TOP1, SEMG1, and PPGB. Several candidate tumor suppressor genes lie outside the common deleted region including SRC, HCK, p107, PTPN1, and CEBP beta. Thirdly, the data allow integration of genetic and physical maps and have refined the map positions of multiple genes. These results will facilitate attempts to identify candidate hematopoietic tumor suppressor genes on 20q.


CH...pi interaction for rhenium-based rectangles: an interaction that is rarely designed into a host-guest pair.
Alkoxy- and thiolato-bridged Re(I) molecular rectangles [{(CO)3Re(mu-ER)2Re(CO)3}2(mu-bpy)2] (ER = SC4H9, 1a; SC8H17, 1b; OC4H9, 2a; OC12H25, 2b; bpy = 4,4'-bipyridine) exhibit strong interactions with several planar aromatic molecules. The nature of their binding was studied by spectral techniques and verified by X-ray diffraction analysis. Standard absorption and fluorescence titrations showed that a relatively strong 1:1 interaction occurs between aromatic guests such as pyrene and these rectangles. The results of a single-crystal X-ray diffraction analysis show that the recognition of 1 with a pyrene molecule is mainly due to CH...pi interactions and the face of the guest pyrene is located over the edges of the bpy linkers of 1. This is a fairly novel example of an interaction that is rarely designed into a host-guest pair. Furthermore, the interaction of 1 with Ag+ results in the self-organization of supramolecular arrays, as revealed by solid-state data.


Writing Abilities in Compulsive Prisoners.
Research has found links between academic failure and criminal offending and suggest that many incarcerated young people have experienced significant behavioral and learning problems in school, which could result in criminal outcomes and poor academic performance. The objective of this study was to analyse writing disorders in impulsive and compulsive prisoners. The sample was composed of 194 male prisoners, of which 81 had been diagnosed with Antisocial Personality Disorder and 113 with Obsessive Compulsive Personality Disorder. Male participants were recruited at the Granada Prison Center. They completed the Demographic, Crime, and Institutional Behavior Interview; the International Personality Disorder Examination (IPDE); The Symptom Checklist (SCL-90-R) and Assessment Battery of Writing Processes (PROESC in its Spanish acronym). We found that prisoners with writing disorders generally have difficulties in the skills necessary to write properly due to impulsive and compulsive behavior.


Suicide in prison: a study of France from 1852 to 1913.
Prison suicide rates in France from 1852 to 1913 were strongly correlated with the overall male suicide rate in France. Measures of domestic social integration were associated with both of these suicide rates, but were less successful in accounting for the prison suicide rate than the overall male suicide rate.


Environmental Regulation, Foreign Direct Investment and Green Technological Progress-Evidence from Chinese Manufacturing Industries.
This study examines the spillover effects of foreign direct investment (FDI) on green technology progress rate (as measured by the green total factor productivity). The analysis utilizes two measures of FDI, labor-based FDI and capital-based FDI, and separately investigates four sets of industry classifications-high/low discharge regulation and high/low emission standard regulation. The results indicate that in the low discharge regulation and low emission standard regulation industry, labor-based FDI has a significant negative spillover effect, and capital-based FDI has a significant positive spillover effect. However, in the high-intensity environmental regulation industry, the negative influence of labor-based FDI is completely restrained, and capital-based FDI continues to play a significant positive green technological spillover effects. These findings have clear policy implications: the government should be gradually reducing the labor-based FDI inflow or increasing stringency of environmental regulation in order to reduce or eliminate the negative spillover effect of the labor-based FDI.


Postoperative daily living activities of geriatric patients administered general or spinal anesthesia for hip fracture surgery: A retrospective cohort study.
Maintaining independence after hip fracture repair is important for geriatric patients and general welfare. We investigated the effects of anesthetic methods on postoperative activities of daily living (ADLs) following hip fracture surgery in elderly patients. The medical records of 12,342 patients aged ≥65 years who underwent typical surgeries for hip fracture using either general anesthesia or spinal anesthesia were reviewed. To adjust for baseline differences and minimize selection bias for the chosen method of anesthesia, patients were matched by propensity scores. Factors affecting the deterioration in ADLs during hospital stay were also investigated in all subjects using a multivariate logistic regression analysis. Eating, grooming, toileting, bathing, and walking were selected as the ADL parameters, as they are considered important for an independent life. Of the 12,342 patients, 6918 (56.1%) received general anesthesia and 5424 (43.9%) received spinal anesthesia. After the propensity score matching, the anesthesia types were not associated with ADL scores except toileting at discharge. Results from the multivariate logistic regression analysis showed that the types of anesthesia were not associated with deterioration in ADL scores. Advanced age, male sex, high Charlson Comorbidity Index scores, psychiatric disease, no administration of nonsteroidal anti-inflammatory drugs, and short length of hospital stay were associated with deterioration in ADL scores. The anesthesia types were not associated with ADL dependency except toileting at discharge. Spinal anesthesia adversely affected toilet use at hospital discharge. However, anesthesia types were not factors that affected deterioration in ADL during hospital stay in elderly patients who underwent hip fracture surgery.


[Quantitative changes in the capillary supply of selected muscles of turkeys, ducks, rats and swine during postnatal development].
The capillary and muscle fibre density, the ratio of capillaries to fibres and the mean diffusion distance were determined in histological cross sections of the M. pectoralis of turkeys and ducks, the M. semitendinosus of rats and the M. longissimus of pigs between hatching or birth and maturity. The data were analyzed by fitting the Janoschek growth curve. The capillary and fibre density decrease asymptomatically, the diffusion distance increases. The M. longissimus of the adult pig is provided with the lowest capillary density resulting in the greatest diffusion distance. The highest capillarity was found in the M. pectoralis of ducks.


CCHCR1-astrin interaction promotes centriole duplication through recruitment of CEP72.
The centrosome is one of the most important non-membranous organelles regulating microtubule organization and progression of cell mitosis. The coiled-coil alpha-helical rod protein 1 (CCHCR1, also known as HCR) gene is considered to be a psoriasis susceptibility gene, and the protein is suggested to be localized to the P-bodies and centrosomes in mammalian cells. However, the exact cellular function of HCR and its potential regulatory role in the centrosomes remain unexplored. We found that HCR interacts directly with astrin, a key factor in centrosome maturation and mitosis. Immunoprecipitation assays showed that the coiled-coil region present in the C-terminus of HCR and astrin respectively mediated the interaction between them. Astrin not only recruits HCR to the centrosome, but also protects HCR from ubiquitin-proteasome-mediated degradation. In addition, depletion of either HCR or astrin significantly reduced centrosome localization of CEP72 and subsequent MCPH proteins, including CEP152, CDK5RAP2, and CEP63. The absence of HCR also caused centriole duplication defects and mitotic errors, resulting in multipolar spindle formation, genomic instability, and DNA damage. We conclude that HCR is localized and stabilized at the centrosome by directly binding to astrin. HCR are required for the centrosomal recruitment of MCPH proteins and centriolar duplication. Both HCR and astrin play key roles in keeping normal microtubule assembly and maintaining genomic stability.


[Nutritional status associated with demographic characteristics in older Peruvian adults].
To describe the nutritional status in older adults and its association with sociodemographic characteristics. A cross-sectional study was conducted. Sampling was probabilistic, stratifi and multistage. Nutritional status was assessed by body mass index (BMI) according to the classifi of nutritional status for the elderly from the Ministry of Health of Peru. The statistical analysis considered the necessary weight for complex samples. The study included 7,267 older adults. 26.8% of participants were underweight, 21.7% overweight, 10.6% obese and 40.8% normal. The average age in the sample was 70.1 ± 8.3 years. Illiteracy (OR 1.9; 95% CI 1.2-3.0), primary education (OR 1.9; 95% CI 1.3-2.9), extreme poverty (OR 2.0; 95% CI 1.6-2.5), living in rural areas (OR 1.8; 95% CI 1.5-2.1), living in the mountains (OR 1.6; 95% CI 1.2-2.2) or jungle (OR 1.6; 95% CI 1.1-2.2) were found associated with underweight. Females (OR 1.8; 95% CI 1.4-2.1), living in urban areas (OR 2.0; 95% CI 1.6-2.5), living in the coastal region (OR 1.5; 95% CI 1.2- 1.8); and not classifi as poor (OR 1.9; 95% CI 1.3-2.9) were associated with overweight. Female sex (OR 3.1; 95% CI 2.3-4.1), primary education (OR 2.4; 95% CI 1.5-4.0) and secondary (OR 2.0; 95% CI 1.2-3.4); live in urban areas (OR 2.2; 95% CI 1.6-2.9), inhabiting the coast (OR 1.8; 95% CI 1.3-2.4), Metro (OR 1.6, 95% CI 1.1-2.2) and jungle (OR 1.6; 95% CI 1.1-2.2), and not classifi as poor (OR 3.5; 95% CI 1.8-7.0) were associated with obesity. The data suggest that both underweight and overweight are common in the elderly population studied.


Triplex-forming oligonucleotide-orthophenanthroline conjugates for efficient targeted genome modification.
The inefficiency of gene modification by homologous recombination can be overcome by the introduction of a double-strand break (DSB) in the target. Engineering the endonucleases needed, however, remains a challenging task that limits widespread application of nuclease-driven gene modification. We report here that conjugates of orthophenanthroline (OP), a DNA cleaving molecule, and triplex-forming oligonucleotides (TFOs), known to bind specific DNA sequences, are synthetic nucleases efficient at stimulating targeted genome modification. We show that in cultured cells, OP-TFO conjugates induce targeted DSBs. An OP-TFO with a unique target was highly efficient, and mutations at the target site were found in approximately 10% of treated cells, including small deletions most likely introduced during DSB repair by nonhomologous end joining. Importantly, we found that when homologous donor DNA was cotransfected, targeted gene modification took place in >1.5% of treated cells. Because triplex-forming sequences are frequent in human and mouse genes, OP-TFO conjugates therefore constitute an important class of site-specific nucleases for targeted gene modification. Harnessing DNA-damaging molecules to predetermined genomic sites, as achieved here, should also provide inroads into mechanisms of DNA repair and cancer.


The glucuronyltransferase B4GAT1 is required for initiation of LARGE-mediated α-dystroglycan functional glycosylation.
Dystroglycan is a cell membrane receptor that organizes the basement membrane by binding ligands in the extracellular matrix. Proper glycosylation of the α-dystroglycan (α-DG) subunit is essential for these activities, and lack thereof results in neuromuscular disease. Currently, neither the glycan synthesis pathway nor the roles of many known or putative glycosyltransferases that are essential for this process are well understood. Here we show that FKRP, FKTN, TMEM5 and B4GAT1 (formerly known as B3GNT1) localize to the Golgi and contribute to the O-mannosyl post-phosphorylation modification of α-DG. Moreover, we assigned B4GAT1 a function as a xylose β1,4-glucuronyltransferase. Nuclear magnetic resonance studies confirmed that a glucuronic acid β1,4-xylose disaccharide synthesized by B4GAT1 acts as an acceptor primer that can be elongated by LARGE with the ligand-binding heteropolysaccharide. Our findings greatly broaden the understanding of α-DG glycosylation and provide mechanistic insight into why mutations in B4GAT1 disrupt dystroglycan function and cause disease.


Mitochondrial DNA structure of an isolated Tunisian Berber population and its relationship with Mediterranean populations.
Douiret is an isolated Berber population from South-Eastern Tunisia. The strong geographic and cultural isolation characterising this population might have contributed to remarkable endogamy and consanguinity, which were practiced for several centuries. The objective of this study is to evaluate the mitochondrial DNA (mtDNA) genetic structure of Douiret and to compare it to other Mediterranean populations with a special focus on major haplogroup T. Genomic DNA was extracted from blood samples of 58 unrelated individuals collected from the different patrilineal lineages of the population. The hypervariable region 1 of the mtDNA was amplified and sequenced. For comparative analyses, additional HVS1 sequences (n = 4857) were compiled from previous studies. The maternal background of the studied sample from Douiret was mainly of Eurasian origin (74%) followed by Sub-Saharan (17%) and North African (3%) lineages. Douiret harbours the highest frequency of haplogroup T in the Mediterranean region, assigned to the unique subclade T1a (38%). Phylogenetic analysis showed an outlier position of Douiret at the Mediterranean level. The genetic structure of Douiret highlights the presence of founders, most likely of Near/Middle Eastern origin, who conquered this area during the Middle/Late Upper Palaeolithic and Neolithic dispersals.


Partially oxidized [2-[(benzoylmethylene)diphenyl-lambda5-phosphino]ethyl]diphenylphosphine as a monohydrate.
The title compound is a co-crystal of [2-[(benzoylmethylene)diphenyl-lambda5-phosphino]ethyl]diphenylphosphine oxide, [2-[(benzoylmethylene)diphenyl-lambda5-phosphino]ethyl]diphenylphosphine and water in an approximate 2:1:3 ratio, with an overall composition of C34H30O1.678P2.H2O. The ylidic portion shows the expected electronic polarization, and the organic components are linked by a combination of C-H...O and C-H...pi(arene) hydrogen bonds.


Convenient method to access new 4,4-dialkoxy- and 4,4-diaryloxy-diaza-s-indacene dyes: Synthesis and spectroscopic evaluation.
A straightforward method for the synthesis of original 4,4-dialkoxy- or 4,4-diaryloxy-diaza-s-indacenes (BODIPY) derivatives obtained by treatment of BODIPY 1 with various alcohols in the presence of AlCl3 is described. The novel compounds are characterized by spectroscopic properties similar to those of the parent BODIPY 1, absorption and emission spectra with similar band shapes, high molar absorption coefficients (epsilon lambda max approximately 80,000 M(-1) cm(-1)), and for most of them high fluorescence quantum yields (Phi exp from 0.52 to 0.71). Among all of the new compounds synthesized, the dye 2 h exhibits higher fluorescence quantum yield (0.71) and lifetime (4.09 ns) than compound 1 and a good chemical stability toward conditions compatible with biological cell-based assays.


From medicine to natural philosophy: Francis Hauksbee's way to the air-pump.
Francis Hauksbee (1660-1713) is well known for his double-barrelled air-pump. However, the origin of this pump, and Hauksbee's background, are often described as a mystery. This text seeks to dispel the riddle. It is argued that Hauksbee's competence as an exceptional maker of air-pumps was developed between 1699 and 1703 as a result of his experiences with the construction, manufacturing and sale of cupping-glasses. His cupping utensils embodied a new design, where syringes were used to evacuate the glasses, instead of the traditional way by fire or mouth suction. These syringes, which in fact were small air-pumps, were perfected between 1699 and 1701. A larger syringe, introduced in 1701, served as a transition from the cupping-syringe to his first air-pump for use in natural philosophy. This syringe was described as a 'combined engine', which could serve as an air-pump, a condensing engine and a syringe for injecting air, wax or mercury into pathological specimens. Hauksbee's first air-pump was a single-barrelled model introduced in 1702, based on the combined engine. Its various features, such as easy and convenient leak-tightening, exact pressure measurements by an in-built barometer and an air-inlet function for readmission of air into the receiver, are discussed. Finally, it is shown that these activities gave Hauksbee the reputation of being an outstanding instrument-maker, years before the double-barrelled air-pump was in sight.


[Supersensitive detection of tyrosine hydroxylase mRNA and neuroblastoma cells by reverse transcription and polymerase chain reaction].
To facilitate the diagnosis of bone marrow metastasis in neuroblastoma, we have developed a method of amplifying and detecting the tyrosine hydroxylase (TH) mRNA sequence in bone marrow cell, using a combination of reverse transcription and the polymerase chain reaction (RT/PCR). By this method, the sequence was detected clearly in the neuroblastoma tissues of all 6 patients and not detected in the bone marrow cells of any of the 9 negative control children. In a reconstitution experiment, one neuroblastoma cell per 100,000 normal bone marrow cells could be detected, thus indicating the great sensitivity of the method. Based on these results, this technique may be of value in the diagnosis and treatment follow-up of bone marrow metastasis of neuroblastoma.


Effects of vitamin D supplementation as an adjuvant therapy in coronary artery disease patients.
Low vitamin D status has been shown to be associated with coronary artery disease. We planned to research the effects of vitamin D3 supplementation on the severity of coronary artery disease. We investigated the effect of 0.5 μg vitamin D3 per day in a randomized, placebo-controlled, double-blind study in 90 stable coronary artery disease patients residing in Beijing. Coronary angiography was performed before and after 6 months of treatment that took place between January and June. 25-Hydroxyvitamin D was measured by chemiluminescence assay. Coronary artery disease severity was assessed by using the SYNTAX scores. In vitamin D supplementation group, there was a significant increase in mean 25-hydroxyvitamin D levels from baseline (19.9 ± 9.8 ng/ml) to 6 months (35.8 ± 12.1 ng/ml; p < 0.001). At 6 months, the primary end point, a difference in the fall of SYNTAX score between the groups was -2.5 (95% CI -5.1 to -0.5; p < 0.001) under intention to treat analysis. Compared with the control group, patients treated with vitamin D3 also had greater decreases in high sensitivity C-reactive protein and renin-angiotensin system activity (p < 0.05). Vitamin D supplementation has beneficial effects on coronary artery disease; it can be an adjuvant therapy for patients with coronary artery disease.


Role of multiple environmental stimuli in control of transcription from a nitrogen-regulated promoter in Escherichia coli with weak or no activator-binding sites.
Nitrogen regulator I (NRI [or NtrC])-phosphate stimulates transcription from the glnAp2 promoter of the glnALG operon in enteric bacteria. Unlike most activators, NRI-phosphate can stimulate transcription without apparent activator binding sites. We observed that when lacZ was controlled by a minimal glnAp2 promoter (without NRI binding sites) in Escherichia coli, lacZ expression was regulated by two different stimuli, the nitrogen status of the medium and the particular amino acid used as a nitrogen source. The latter stimulus did not affect the activity of the wild-type glnAp2 promoter, which has two high-affinity NRI binding sites. We present several lines of evidence that suggest that the concentration of NRI-phosphate limits the activity of the minimal glnAp2 promoter in vivo. Our results also suggest that nitrogen regulator II-dependent phosphorylation of NRI cannot account for the proposed variations in the concentration of NRI-phosphate. Therefore, to account for the regulation of the minimal glnAp2 promoter by two environmental stimuli, we propose that at least two protein kinases phosphorylate NRI during nitrogen-limited growth. We isolated and characterized mutants in which NRI could not stimulate transcription from the minimal glnAp2 promoter but could activate transcription from the wild-type glnAp2 promoter. These mutants could not utilize arginine or proline as a nitrogen source, suggesting that degradation of some nitrogen sources may require transcription from promoters similar to the minimal glnAp2 promoter.


[Echo Doppler post total hip arthroplasty: retrospective: study of 111 patients at CHBAH].
We studied all the patients who underwent a total hip arthroplasty during the last six months of 2001. They all had a postoperative venous Doppler echo (possibly repeated) followed by a phlebography in case of clinical doubt. They all received a prophylactic treatment by low molecular weight heparin. Our rate of deep venous thrombosis was 22% (from which 3% are) and of pulmonary emboli was 2,7%. These figures are in keeping with those of large controlled studies. The risk factors identified are age and female gender. We discuss, based on the literature, the significance of lower deep venous thrombosis. We also discuss the best strategy to detect postoperative deep venous thrombosis in order to improve the cost/effectiveness ratio.


The impact of various work schedules on sleep complaints and minor accidents during work or leisure time: evidence from a national survey.
To explore the prevalence of insomnia symptoms and excessive daytime sleepiness (EDS) in different work schedules and the impact of shift schedules on the risk of minor accidents during work or leisure time. Using the data from Taiwan Social Development Trend Survey in 2005 (n = 18,794), insomnia symptoms, EDS, and minor accidents were analyzed using descriptive statistics and multivariable logistic regression model. The evening-to-night group had significantly higher prevalence rates of insomnia symptoms. Higher prevalence rate of EDS was presented in both day-to-evening and evening-to-night groups. Adjusting for confounders, the day-to-evening shift had a higher odds ratio of minor accidents as compared with the fixed daytime workers. Extended-shift workers tend to experience insomnia symptoms and EDS and have an increased likelihood of minor accidents.


A Retrospective Study of Clinical Profile and Long Term Outcome to Imatinib Mesylate Alone in Childhood Chronic Myeloid Leukemia in Chronic Phase.
Chronic myeloid leukemia (CML) is relatively rare malignancy in childhood. There are limited studies of use of Imatinib Mesylate (IM) alone in management of CML in this age group. We retrospectively analyzed the outcome of 30 consecutive children with CML chronic phase treated with IM alone. The median age at the time of diagnosis was 11 years with male preponderance. Asthenia and abdominal discomfort due to splenomegaly were the most common presenting features and splenomegaly a dominant sign. At time of starting IM, 19 children were in early CP while 11 were in late CP. Complete hematological remission was achieved in 90% with a median time to achieve CHR was 60 days. Among evaluable children, 83.3% achieved cytogenetic response (CyGR). Those who achieved complete CyGR were in major molecular remission at the time of last follow up. At 3 years, progression-free survival was 81.5% and overall survival was 100%. At 10 years, 12 (40%) children failed on IM therapy of which 3(10%) children developed primary IM resistance while 9 (30%) developed secondary IM resistance. IM was well tolerated and severe (grade III-IV) events were infrequent. Non-haematological toxicities were uncommon except hypopigmentation of skin which was seen in 60% of the cohort. Presenting features of CML-CP in children is comparable to other Indian and international studies. IM is very effective and safe drug for the first line treatment of CML-CP in children. It is very effective in inducing CHR. Adherence to treatment is very important for achieving CyGR and long term survival. This data will be useful for financially deprived children in developing countries where allogeneic stem cell transplant (SCT) or second line tyrosine kinase inhibitors (TKIs) is not an affordable option.


A mass spectrometry assay for detection of endogenous and lentiviral engineered hemoglobin in cultured cells and sickle cell disease mice.
Sickle cell disease (SCD) results from a sequence defect in the β-globin chain of adult hemoglobin (HbA) leading to expression of sickle hemoglobin (HbS). It is traditionally diagnosed by cellulose-acetate hemoglobin electrophoresis or high-performance liquid chromatography. While clinically useful, these methods have both sensitivity and specificity limitations. We developed a novel mass spectrometry (MS) method for the rapid, sensitive and highly quantitative detection of endogenous human β-globin and sickle hβ-globin, as well as lentiviral-encoded therapeutic hβAS3-globin in cultured cells and small quantities of mouse peripheral blood. The MS methods were used to phenotype homozygous HbA (AA), heterozygous HbA-HbS (AS) and homozygous HbS (SS) Townes SCD mice and detect lentiviral vector-encoded hβAS3-globin in transduced mouse erythroid cell cultures and transduced human CD34+ cells after erythroid differentiation. hβAS3-globin was also detected in peripheral blood 6 weeks post-transplant of transduced Townes SS bone marrow cells into syngeneic Townes SS mice and persisted for over 20 weeks post-transplant. As several genome-editing and gene therapy approaches for severe hemoglobin disorders are currently in clinical trials, this MS method will be useful for patient assessment before treatment and during follow-up.


Policy makers' perspectives on the utility of a national study of child maltreatment.
Reliable national child maltreatment data are needed for developing and modifying policies aimed at preventing child maltreatment and helping child victims of maltreatment. Health Canada hosted a daylong forum in 2002 to solicit feedback from senior Government of Canada policy and program officials involved in child welfare programs and research in regard to the data collected in the Canadian Incidence Study of Reported Child Abuse and Neglect (CIS). This article reviews the discussions and debates regarding the utility of the CIS data for government policy makers and reflects on the implications for surveillance and knowledge in the area of child maltreatment. The key themes are definitions and measurement issues, the value of enhanced and additional data, and challenges to linking research and practice.


Atrioventricular nodal ablation and biventricular pacing therapy with coronary venoplasty for severe heart failure with drug refractory atrial tachycardia.
A 53-year-old man was admitted to our hospital for dyspnea, associated with atrial tachycardia. He underwent mitral valve replacement and Maze operation for mitral regurgitation and atrial fibrillation and since then he suffered from drug refractory atrial tachycardia followed by cardiogenic shock with systolic heart failure. Atrial tachycardia with rapid ventricular response was medically refractory, and radical catheter ablation was thought to be very difficult due to post Maze operation, mitral mechanical valve replacement and unstable hemodynamics. Thus, atrioventricular nodal ablation, which was s safe procedure compared to the radical ablation for this patient, and biventricular pacemaker implantation, which required coronary venoplasty, were performed. Combination therapy may be one of the treatments for heart failure patients with drug refractory tachyarrhythmias.


Operator-as-a-Consumer: A Novel Energy Storage Sharing Approach Under Demand Charge.
Energy storage systems (ESSs)-based demand response (DR) is an appealing way to save electricity bills for consumers under demand charge and time-of-use (TOU) price. In order to counteract the high investment cost of ESS, a novel operator-enabled ESS sharing scheme, namely, the "operator-as-a-consumer (OaaC)," is proposed and investigated in this article. In this scheme, the users and the operator form a Stackelberg game. The users send ESS orders to the operator and apply their own ESS dispatching strategies for their own purposes. Meanwhile, the operator maximizes its profit through optimal ESS sizing and scheduling, as well as pricing for the users' ESS orders. The feasibility and economic performance of OaaC are further analyzed by solving a bilevel joint optimization problem of ESS pricing, sizing, and scheduling. To make the analysis tractable, the bilevel model is first transformed into its single-level mathematical program with equilibrium constraints (MPEC) formulation and is then linearized into a mixed-integer linear programming (MILP) problem using multiple linearization methods. Case studies with actual data are utilized to demonstrate the profitability for the operator and simultaneously the ability of bill saving for the users under the proposed OaaC scheme.


Diurnal Variation Has Effect on Differential Gene Expression Analysis in the Hippocampus of the Pilocarpine-Induced Model of Mesial Temporal Lobe Epilepsy.
The molecular mechanisms underlying epileptogenesis have been widely investigated by differential gene expression approach, especially RT-qPCR methodology. However, controversial findings highlight the occurrence of unpredictable sources of variance in the experimental designs. Here, we investigated if diurnal rhythms of transcript's levels may impact on differential gene expression analysis in hippocampus of rats with experimental epilepsy. For this, we have selected six core clock genes (Per1, Per3, Bmal1, Clock, Cry1 and Cry2), whose rhythmic expression pattern in hippocampus had been previously reported. Initially, we identified Tubb2a/Rplp1 and Tubb2a/Ppia as suitable normalizers for circadian studies in hippocampus of rats maintained to 12:12 hour light:dark (LD) cycle. Next, we confirmed the temporal profiling of Per1, Per3, Bmal1, Cry1 and Cry2 mRNA levels in the hippocampus of naive rats by both Acrophase and CircWave statistical tests for circadian analysis. Finally, we showed that temporal differences of sampling can change experimental results for Per1, Per3, Bmal1, Cry1 and Cry2, but not for Clock, which was consistently decreased in rats with epilepsy in all comparison to the naive group. In conclusion, our study demonstrates it is mandatory to consider diurnal oscillations, in order to avoid erroneous conclusions in gene expression analysis in hippocampus of rats with epilepsy. Investigators, therefore, should be aware that genes with circadian expression could be out of phase in different animals of experimental and control groups. Moreover, our results indicate that a sub-expression of Clock may be involved in epileptogenicity, although the functional significance of this remains to be investigated.


LPMO-supported saccharification of biomass: effects of continuous aeration of reaction mixtures with variable fractions of water-insoluble solids and cellulolytic enzymes.
High substrate concentrations and high sugar yields are important aspects of enzymatic saccharification of lignocellulosic substrates. The benefit of supporting the catalytic action of lytic polysaccharide monooxygenase (LPMO) through continuous aeration of slurries of pretreated softwood was weighed against problems associated with increasing substrate content (quantitated as WIS, water-insoluble solids, in the range 12.5-17.5%), and was compared to the beneficial effect on the saccharification reaction achieved by increasing the enzyme preparation (Cellic CTec3) loadings. Aerated reactions were compared to reactions supplied with N2 to assess the contribution of LPMO to the saccharification reactions. Analysis using 13C NMR spectroscopy, XRD, Simons' staining, BET analysis, and SEM analysis was used to gain further insights into the effects of the cellulolytic enzymes on the substrate under different reaction conditions. Although glucose production after 72 h was higher at 17.5% WIS than at 12.5% WIS, glucan conversion decreased with 24% (air) and 17% (N2). Compared to reactions with N2, the average increases in glucose production for aerated reactions were 91% (12.5% WIS), 70% (15.0% WIS), and 67% (17.5% WIS). Improvements in glucan conversion through aeration were larger (55-86%) than the negative effects of increasing WIS content. For reactions with 12.5% WIS, increased enzyme dosage with 50% improved glucan conversion with 25-30% for air and N2, whereas improvements with double enzyme dosage were 30% (N2) and 39% (air). Structural analyses of the solid fractions revealed that the enzymatic reaction, particularly with aeration, created increased surface area (BET analysis), increased disorder (SEM analysis), decreased crystallinity (XRD), and increased dye adsorption based on the cellulose content (Simons' staining). The gains in glucan conversion with aeration were larger than the decreases observed due to increased substrate content, resulting in higher glucan conversion when using aeration at the highest WIS value than when using N2 at the lowest WIS value. The increase in glucan conversion with double enzyme preparation dosage was smaller than the increase achieved with aeration. The results demonstrate the potential in using proper aeration to exploit the inherent capacity of LPMO in enzymatic saccharification of lignocellulosic substrates and provide detailed information about the characteristics of the substrate after interaction with cellulolytic enzymes.


Engineering the vibrational coherence of vision into a synthetic molecular device.
The light-induced double-bond isomerization of the visual pigment rhodopsin operates a molecular-level optomechanical energy transduction, which triggers a crucial protein structure change. In fact, rhodopsin isomerization occurs according to a unique, ultrafast mechanism that preserves mode-specific vibrational coherence all the way from the reactant excited state to the primary photoproduct ground state. The engineering of such an energy-funnelling function in synthetic compounds would pave the way towards biomimetic molecular machines capable of achieving optimum light-to-mechanical energy conversion. Here we use resonance and off-resonance vibrational coherence spectroscopy to demonstrate that a rhodopsin-like isomerization operates in a biomimetic molecular switch in solution. Furthermore, by using quantum chemical simulations, we show why the observed coherent nuclear motion critically depends on minor chemical modifications capable to induce specific geometric and electronic effects. This finding provides a strategy for engineering vibrationally coherent motions in other synthetic systems.


ClearT: a detergent- and solvent-free clearing method for neuronal and non-neuronal tissue.
We describe a clearing method for enhanced visualization of cell morphology and connections in neuronal and non-neuronal tissue. Using Clear(T) or Clear(T2), which are composed of formamide or formamide/polyethylene glycol, respectively, embryos, whole mounts and thick brain sections can be rapidly cleared with minimal volume changes. Unlike other available clearing techniques, these methods do not use detergents or solvents, and thus preserve lipophilic dyes, fluorescent tracers and immunohistochemical labeling, as well as fluorescent-protein labeling.


Recruitment activities and sociodemographic factors that predict attendance at a mammographic screening program.
A random sample of 2266 women aged 50 to 69 years was used to investigate factors that predict attendance at a free Australian mammographic screening program. The most important predictor was receipt of a personal invitation letter. A letter that included an appointment time increased attendance 132-fold initially and decreased to 20 times baseline after 14 days. A letter that did not include an appointment time increased attendance 12-fold, and a second letter to nonattenders increased attendance approximately 13-fold. Attendance declined with increasing distance from the program and with increases in the percentage of non-English speaking women in a neighborhood, but was higher in areas of higher socioeconomic status.