Brain atrophy in a murine model of chronic fatigue syndrome and beneficial effect of Hochu-ekki-to (TJ-41).
Brain-derived neurotrophic factor (BDNF) is associated with the main symptoms of chronic fatigue syndrome (CFS) and neuron apoptosis. Nevertheless, no study has been performed directly to explore the relationship between CFS, BDNF and neuron apoptosis. We induced a CFS model by six injections of killed Brucella abortus antigen in BALB/c mice and treated them with Hochu-ekki-to (TJ-41). Daily running activity, body weight (BW), ratio of cerebral weight to BW (CW/BW) and expression levels of BDNF and Bcl-2 mRNA in the hippocampus were determined. The daily activity and CW/BW decreased significantly in the CFS model. BDNF and Bcl-2 mRNA expression levels in the hippocampus were suppressed in the CFS model and TJ-41 treated mice, while no significant difference was found between them. We improved a murine model to investigate the relationship between CFS and brain dysfunction. In this model, reduced daily activity might have been associated with decreased hippocampal BDNF mRNA expression, hippocampal apoptosis and brain atrophy. TJ-41 increased the daily running activity of the model, which was independent of brain recovery.


Lysosomal enzymes in preterm infants with bronchopulmonary dysplasia: a potential diagnostic marker.
Some lysosomal glycohydrolases (N-acetyl-beta-D-glucosaminidase and their major isoenzymes, beta-D-glucuronidase, alpha-D-galactosidase, beta-D-galactosidase and alpha-D-glucosidase) were investigated in the plasma of 36 preterm infants with respiratory distress, 11 of whom developed bronchopulmonary dysplasia (BPD), in order to evaluate the role of the lysosomal apparatus in the disease. Enzyme activity was assayed fluorimetrically; the major N-acetyl-beta-D-glucosaminidase (NAG) isoenzymes were separated using a routine chromatofocusing procedure; the diagnostic efficiency was evaluated by Bayes theorem. The mean levels of almost all glycohydrolases considered were significantly higher in BPD than in non-BPD infants. Among NAG major isoenzymes, an increase was found only in form A. No variation was evident in the plasma levels of glycohydrolases during dexamethasone therapy. Data from a retrospective analysis performed in all preterms considered, show that alpha-D-galactosidase and beta-D-galactosidase differentiate a posteriori BPD and non-BPD subjects. These enzymes, after a priori verification of their diagnostic potential in preterm infants at risk of BPD development, could acquire an important predictive value.


Association between Dietary Fiber Intake and Colorectal Adenoma.
Dietary fiber intake has been suggested to decrease the risks of colorectal cancer (CRC) and adenoma. This cross-sectional study aimed to evaluate the association between total dietary fiber intake and colorectal adenoma among asymptomatic Korean adults. Individuals who received a screening colonoscopy between May and December of 2011 were recruited. Multivariable-adjusted logistic regression was used to assess the odds ratios (ORs) and 95% confidence intervals (CIs) of colorectal adenoma. 558 of the 1,716 study participants were diagnosed with colorectal adenoma. No significant association between total dietary fiber intake and colorectal adenoma was found; ORs (95% CIs) for subsequent quintiles compared to the bottom quintile were 1.00 (0.69-1.46), 1.11 (0.73-1.71), 0.97 (0.57-1.65), and 0.88 (0.46-1.71; P for trend = 0.65). Dietary fiber intakes from cereal, fruit, vegetable, or legume weren't associated with colorectal adenoma. When we compared >30 g/d to ≤10 g/d of total dietary fiber intake, OR (95% CI) was 0.32 (0.10-1.02; P for trend = 0.23). In the analyses of advanced or high-risk state and location of adenoma, we didn't observe significant associations. In conclusion, dietary fiber intake was not associated with colorectal adenoma in Korean adults. However, the association for low intake of dietary fiber warrants further investigation.


Quantification of Plasmodium falciparum HRP-2 as an alternative method to [3H]hypoxanthine incorporation to measure the parasite reduction ratio in vitro.
In the absence of a highly efficacious vaccine, chemotherapy remains the cornerstone to control malaria morbidity and mortality. The threat of the emergence of parasites resistant to artemisinin-based combination therapies highlights the need for new antimalarial drugs ideally with superior properties. The killing rate reflects the speed of action of antimalarial drugs, which can be measured in vitro through the parasite reduction ratio (PRR) assay to shortlist interesting candidates. As a standard, the in vitro PRR assay is performed by measuring [3H]hypoxanthine incorporation of Plasmodium falciparum. This methodology is restricted to specialised laboratories owing to the handling of radioactive material. In this work, we describe a sandwich enzyme-linked immunosorbent assay to detect P. falciparum histidine-rich protein 2 (HRP-2) as an alternative methodology to assess the PRR. We first validated the methodology with established antimalarial drugs (artesunate, chloroquine, pyrimethamine and atovaquone) by comparing our results with previous results of the [3H]hypoxanthine incorporation readout provided by an expert laboratory, and subsequently assessed the speed of action of four new antimalarial candidates (compound 22, chlorotonil A, boromycin and ivermectin). The HRP-2 PRR assay achieved comparable results to the [3H]hypoxanthine incorporation readout in terms of parasite growth rate over time, lag phase and parasite clearance time. In addition, parasite growth following drug exposure was quantified after 7, 14, 21 and 28 days of recovery time. In conclusion, the PRR assay based on HRP-2 is similar to [3H]hypoxanthine in determining a drug's parasite killing rate and can be widely used in all research laboratories.


Nore1 inhibits tumor cell growth independent of Ras or the MST1/2 kinases.
Nore1, a noncatalytic protein identified by its ability to bind selectively to active Ras, is most closely related in amino-acid sequence to the tumor suppressor RASSF1. Both are expressed predominantly as a longer (Nore1A/RASSF1A) and/or shorter (Nore1B/RASSF1C) polypeptide; all four polypeptides contain a Ras-association domain and bind, through their conserved carboxytermini, the proapoptotic protein kinases MST1 and MST2. Moreover, the expression of the longer polypeptide is downregulated in human tumor cell lines through promoter methylation (frequently for RASSF1A, less regularly for Nore1A). Forced expression of RASSF1A in several such lines (including the NSCLC line A549) has been shown to suppress tumorigenicity; herein we inquire whether Nore has growth inhibitory activity. Four tumor cell lines were tested, selected for their low expression of both Nore1A and Nore1B; the two NSCLC lines, A549 and NCI-H460, each have a mutant active Ras oncogene, whereas the two melanoma lines G361 and M14 each contain the constitutively active BRaf(V599E) oncogene and wild-type Ras. The expression of Nore1A or Nore1B suppresses colony formation by the A549 and G361 lines, as effectively in A549 as does RASSF1A; colony formation in the NCI-H460 and M14 lines is unaffected. Nore1A inhibits anchorage-independent growth by A549 cells and delays A549 progression through G1 without evidence of increased apoptosis. The growth suppressive action of Nore1A is largely unaffected by deletion of both the MST- and Ras-binding domains, as well as by mutation of the Nore1A zinc finger. Thus, Nore1 suppresses the growth of some tumor cell lines through as yet unidentified effectors, independent of Ras-like proteins or MST1/2.


Evaluation of the chemical profile from four germplasms sources of Pruni Semen using UHPLC-LTQ-Orbitrap-MS and multivariate analyses.
Pruni Semen, the seed of several unique Prunus plants, is a traditional purgative herbal material. To determine the authentic sources of Pruni Semen, 46 samples from four species were collected and analyzed. Ten compounds including multiflorin A (Mul A), a notable purative compound, were isolated and identified by chemical separation and nuclear magnetic resonance spectroscopy. Seventy-six communal components were identified by ultra-high performance liquid chromatography with linear ion trap-quadrupole Orbitrap mass spectrometry, and acetyl flavonoid glycosides were recognized as characteristic constituents. The flavonoids were distributed in the seed coat and cyanogenic glycosides in the kernel. Based on this, methods for identifying Pruni Semen from different sources were established using chemical fingerprinting, quantitative analysis of the eight principal compounds, hierarchical cluster analysis, principal component analysis, and orthogonal partial least squares discriminant analysis. The results showed that the samples were divided into two categories: one is the small seeds from Prunus humilis (Ph) and Prunus japonica (Pj), and the other is the big seeds from Prunus pedunculata (Pp) and Prunus triloba (Pt). The average content of Mul A was 3.02, 6.93, 0.40, and 0.29 mg/g, while the average content of amygdalin was 18.5, 17.7, 31.5, and 30.9 mg/g in Ph, Pj, Pp, and Pt, respectively. All the above information suggests that small seeds might be superior sources of Pruni Semen. This is the first comprehensive report on the identification of chemical components in Pruni Semen from different species.


Inhibition of interleukin 2 production by prostaglandin E2 is not absolute but depends on the strength of the stimulating signal.
In view of the eminently important role of interleukin-2 (IL-2) in T-cell responses, and in view of reports about immune stimulatory effects of PGE2, we reinvestigated the question whether PGE2 inhibits IL-2 production. It was found that PGE2 does not inhibit IL-2 production in murine spleen cell cultures after optimal stimulation (5 micrograms/ml concanavalin A) but does inhibit at suboptimal stimulation conditions. The failure of PGE2 to inhibit IL-2 production at optimal concanavalin A concentration was demonstrated by two independent IL-2 assays namely by the co-stimulator assay and by the proliferation of IL-2-dependent T-cell clone W-2. Our observations indicated that the inhibitory effect of PGE2 depends on the strength of the stimulating signal. IL-2 production in cultures with 5 micrograms/ml concanavalin A was also not suppressed by PGE1, by prostaglandin D2, thromboxane B2 (T X B2), and prostaglandin F2.


A Metabolomics Study of Hypoxia Ischemia during Mouse Brain Development Using Hyperpolarized 13C.
Hyperpolarized 13C spectroscopic magnetic resonance spectroscopy (MRS) is an advanced imaging tool that may provide important real-time information about brain metabolism. Mice underwent unilateral hypoxia-ischemia (HI) on postnatal day (P)10. Injured and sham mice were scanned at P10, P17, and P31. We used hyperpolarized 13C MRS to investigate the metabolic exchange of pyruvate to lactate in real time during brain development following HI. 13C-1-labeled pyruvate was hyperpolarized and injected into the tail vein through a tail-vein catheter. Chemical-shift imaging was performed to acquire spectral-spatial information of the metabolites in the brain. A voxel placed on each of the injured and contralateral hemispheres was chosen for comparison. The difference in pyruvate delivery and lactate to pyruvate ratio was calculated for each of the voxels at each time point. The normalized lactate level of the injured hemisphere was also calculated for each mouse at each of the scanning time points. There was a significant reduction in pyruvate delivery and a higher lactate to pyruvate ratio in the ipsilateral (HI) hemisphere at P10. The differences decreased at P17 and disappeared at P31. The normalized lactate level in the injured hemisphere increased from P10 to P31 in both sham and HI mice without brain injury. We describe a method for detecting and monitoring the evolution of HI injury during brain maturation which could prove to be an excellent biomarker of injury.


Developing decision support systems in clinical bioinformatics.
There is a growing demand for tools to support clinicians utilize genomic results generated by molecular diagnostic and cytogenetic methods in support of their decision-making. This chapter reviews existing experience and methods for the design, implementation and evaluation of clinical bioinformatics electronic decision support systems (EDSS). It provides a roadmap for identifying decision tasks for automation and selecting optimal tools for building task-specific systems. Key success factors for EDSS implementation and evaluation are also outlined.


Dietary iron regulates intestinal goblet cell function and alleviates Salmonella typhimurium invasion in mice.
Iron is an important micronutrient that plays a vital role in host defenses and bacterial pathogenicity. As iron treatments increase the risk of infection by stimulating the growth and virulence of bacterial pathogens, their roles in anti-infection immunity have frequently been underestimated. To estimate whether adequate dietary iron intake would help defend against pathogenic bacterial infection, mice were fed iron-deficient (2 mg kg-1 feed), iron-sufficient (35 mg kg-1 feed), or iron-enriched diet (350 mg kg-1 feed) for 12 weeks, followed by oral infection with Salmonella typhimurium. Our results revealed that dietary iron intake improved mucus layer function and decelerated the invasion of the pathogenic bacteria, Salmonella typhimurium. Positive correlations between serum iron and the number of goblet cells and mucin2 were found in response to total iron intake in mice. Unabsorbed iron in the intestinal tract affected the gut microbiota composition, and the abundance of Bacteroidales, family Muribaculaceae, was positively correlated with their mucin2 expression. However, the results from antibiotic-treated mice showed that the dietary iron-regulated mucin layer function was not microbial-dependent. Furthermore, in vitro studies revealed that ferric citrate directly induced mucin2 expression and promoted the proliferation of goblet cells in both ileal and colonic organoids. Thus, dietary iron intake improves serum iron levels, regulates goblet cell regeneration and mucin layer function, and plays a positive role in the prevention of pathogenic bacteria.


Re-investigation of functional gastrointestinal disorders utilizing a machine learning approach.
Functional gastrointestinal disorders (FGIDs), as a group of syndromes with no identified structural or pathophysiological biomarkers, are currently classified by Rome criteria based on gastrointestinal symptoms (GI). However, the high overlap among FGIDs in patients makes treatment and identifying underlying mechanisms challenging. Furthermore, disregarding psychological factors in the current classification, despite their approved relationship with GI symptoms, underlines the necessity of more investigation into grouping FGID patients. We aimed to provide more homogenous and well-separated clusters based on both GI and psychological characteristics for patients with FGIDs using an unsupervised machine learning algorithm. Based on a cross-sectional study, 3765 (79%) patients with at least one FGID were included in the current study. In the first step, the clustering utilizing a machine learning algorithm was merely executed based on GI symptoms. In the second step, considering the previous step's results and focusing on the clusters with a diverse combination of GI symptoms, the clustering was re-conducted based on both GI symptoms and psychological factors. The first phase clustering of all participants based on GI symptoms resulted in the formation of pure and non-pure clusters. Pure clusters exactly illustrated the properties of most pure Rome syndromes. Re-clustering the members of the non-pure clusters based on GI and psychological factors (i.e., the second clustering step) resulted in eight new clusters, indicating the dominance of multiple factors but well-discriminated from other clusters. The results of the second step especially highlight the impact of psychological factors in grouping FGIDs. In the current study, the existence of Rome disorders, which were previously defined by expert opinion-based consensus, was approved, and, eight new clusters with multiple dominant symptoms based on GI and psychological factors were also introduced. The more homogeneous clusters of patients could lead to the design of more precise clinical experiments and further targeted patient care.


[One-year results on the safety and efficacy of the InnFocus MicroShunt™ depending on placement and concentration of mitomycin C].
To compare intraocular pressure (IOP) at one year with the InnFocus MicroShunt(®) with or without cataract surgery with according to placement and concentration of mitomycin C (MMC) DESIGN: A retrospective two-center, two-surgeon study (France and Dominican Republic). Adults with POAG requiring filtering surgery. One MicroShunt(®) was placed in one eye of each patient. The effect of concentration and site of application of MMC was assessed by IOP and medication reduction at one year. Eighty-seven eyes were studied with one-year follow-up. Twenty-three eyes treated with 0.4 mg/mL MMC close to the limbus demonstrated a 55% reduction in IOP from 23.8 ± 5.3 at baseline to 10.7 ± 2.8 mmHg at one year. Topical glaucoma medication/patient was reduced 85% from 2.4 ± 0.9 to 0.3 ± 0.8. Thirty-one eyes treated with 0.2mg/mL MMC close to the limbus demonstrated a 52% reduction in IOP from 27.9 ± 6.7 at baseline to 13.3 ± 3.3 mmHg at one year. Topical glaucoma medication/patient was reduced 88% from 2.5 ± 1.4 to 0.5 ± 1.0. Thirty-three eyes treated with 0.4 mg/mL MMC deep in the pocket demonstrated a 38% reduction in IOP from 25.4 ± 7.9 at baseline to 15.7 ± 4.6 mmHg at one year. Topical glaucoma medication/patient was reduced 72% from 2.9 ± 1.0 to 0.8 ± 1.3. There were no sight-threatening long-term adverse events. The InnFocus MicroShunt(®) is a filtering surgery whose efficacy is related to the location of application and concentration of MMC used.


The ferric iron chelator 2,2'-dipyridyl attenuates basilar artery vasospasm and improves neurological function after subarachnoid hemorrhage in rabbits.
We investigated the efficacy of the ferrous iron (Fe(2+)) chelator 2,2'-dipyridyl (DP) to attenuate cerebral vasospasm after subarachnoid hemorrhage (SAH). Thirty-six New Zealand white rabbits were randomly assigned to four groups: untreated control, SAH, SAH + dimethyl sulfoxide (DMSO) vehicle, and SAH + DP. SAH was induced by injection of autologous blood into the cisterna magna and then DP or vehicle was infused into the cistern magna for 5 days (20 mg/kg/day or an equal volume of DMSO). Neurological deficit score (NDS) was used to assess neurological function and cerebral angiography to measure basilar artery (BA) diameter following SAH. TUNEL staining was used to detect BA endothelial cell apoptosis, and immunohistochemistry and Western blotting to assess changes in caspase-3 protein levels 5 days post-SAH. The SAH + DP group had a significantly larger mean BA diameter and lower mean NDS post-SAH compared to the SAH + DMSO and SAH groups (p < 0.05). TUNEL-positive cell numbers and caspase-3 levels were significantly reduced in BA endothelial cells of the SAH + DP group as compared to the SAH and SAH + DMSO groups (p < 0.05). The iron chelator DP reduced vasospasm and neurological sequelae in rabbits, likely by chelating the Fe(2+) in oxyhemoglobin and reducing oxidative stress-induced endothelial cell apoptosis.


A novel, direct, reagent-free method for the detection of beeswax adulteration by single-reflection attenuated total reflectance mid-infrared spectroscopy.
In this work, a novel, direct, reagent-free method for the detection of beeswax adulteration by paraffin, microcrystalline wax, tallow and stearic acid using single-reflection attenuated total reflectance mid-infrared spectroscopy was developed. The use of the absorbance ratios of [Formula: see text] , [Formula: see text] and [Formula: see text] allows a minimum of 5% paraffin/microcrystalline wax and tallow adulteration and 0.5% stearic acid adulteration of beeswax to be detected. The upper and lower critical limits for beeswax authenticity were established from the analysis of virgin beeswax and were validated by independent analysis of real sheet and comb beeswax samples using high-temperature gas chromatography with flame-ionization detection. In addition to its simplicity with respect to sample handling, the amount of sample and the time needed are far less than those required in previously described methods, which are based on chemical analysis and chromatographic techniques. These advantages result in time and cost savings, an increase in the number of samples that can be analyzed, and, most importantly, the detection of the main beeswax adulterants using a single method.


Pre-hospital use of direct oral anticoagulants agents is associated with a lower risk of major bleeding events in critically ill patients: A single academic center experience.
The last decade has witnessed significant advancements in direct oral anticoagulants (DOACs), transforming the landscape of anticoagulation therapy. With the uptrend in DOACs use, critical care physicians are encountering more patients with pre-hospital DOACs prescription. Safety and real world outcomes-related data on DOACs use in critically ill patients are scarce. We assess the risk of major bleeding (MB) events and patient-centered outcomes with pre-hospital use of direct oral anticoagulant agents (DOACs) compared to warfarin therapy. Observational study in a single large academic center from January 1st, 2012, through May 4th, 2018. We included adult critically ill patients with warfarin or one of the DOACs, as active medications at the time of hospital admission. The primary outcome was major bleeding (MB), based on the ISTH criteria RESULTS: 99,481 patients were screened; 558 and 3037 patients were included in the final analysis for the DOAC and warfarin groups, respectively. Multivariable analysis showed that the pre-hospital use of DOACs was associated with lower odds for major bleeding events, GI bleeding, need for endoscopic intervention, hemorrhagic shock, any blood transfusion; but higher odds of intracranial bleeding, as compared to warfarin use. There was no difference in hospital length of stay or ICU-free days. Pre-hospital use of DOACs among critically ill patients is associated with lower major bleeding events, GI bleeding, need for endoscopic intervention, and blood transfusion but a higher risk for intracranial bleeding.


Gallium-transferrin as a macromolecular tracer of vascular permeability.
Labeling of plasma transferrin with gallium was investigated to determine whether the gallium-transferrin complex could be effectively used as a macromolecular tracer in studies of capillary permeability using Positron Emission Tomography (PET). Three gallium-plasma preparations were tested and 2 h biodistribution studies were performed in rats. The three preparations gave similar blood clearance and tissue distribution data, but the methods used for evaluating gallium-transferrin binding were found to be suboptimal. Gallium clearance from blood was biexpoential with both components faster than that of 125I-albumin. Gallium distribution spaces in all tissues including intracerebral Walker-256 tumors were larger than those of albumin. These results indicate a relative instability of the gallium-transferrin complex in vivo, which appears to preclude its use as an acceptable radiolabeled protein for vascular permeability studies using PET.


Adenovirus vectors activate Vδ2+ γδT cells in a type I interferon-, TNF-, and IL-18-dependent manner.
Vδ2+ γδT cells are unconventional T cells that can be activated by cytokines without TCR signaling. Adenovirus vaccine vectors activated Vδ2+ γδT cells in an interleukin 18-, TNF-, and type I interferon-dependent manner. This stimulatory capacity was associated with adenovirus vectors of non-species C origin, including the ChAdOx1 vaccine platform.


Material selection for landfill leachate piping by using a grey target decision-making approach.
Landfill leachate, due to its quantity and inherent risk, is generally collected and transported by piping system for advanced treatment. During the piping, the pipe materials may react with leachate, resulting in corrosion and scaling. In order to reduce possible failures and mitigate the associated consequences, this study provides an indicator system for material selection to aid the pipe system design. The material functional, economic, and environmental attributes are incorporated into the indicator system, to perform a precise selection of commercial drainage pipe materials, thus improving empirically oriented selection. Four common drainage pipe materials including high-density polyethylene (HDPE), polyvinyl chloride (PVC), galvanized steel, and seamless steel are taken as the material alternatives for the selection. Based upon their experimental data, a grey target decision-making framework is employed to perform the priority ranking of the materials. The results indicate that HDPE has the best performance, followed by PVC, galvanized steel, and seamless steel. This study discusses the validity of the selection results and the applicability of the proposed method, to provide insight into leachate piping system design.


Post-translational modifications induced by nitric oxide (NO): implication in cancer cells apoptosis.
Post-translational modifications of proteins can regulate the balance between survival and cell death signals. It is increasingly recognized that nitric oxide (NO) and reactive oxygen species (ROS)-induced post-translational modifications could play a role in cell death. This review provides an introduction of current knowledge of NO proteins modifications promoting or inhibiting cell death with special attention in cancer cells.


Crystallographic snapshots of tyrosine phenol-lyase show that substrate strain plays a role in C-C bond cleavage.
The key step in the enzymatic reaction catalyzed by tyrosine phenol-lyase (TPL) is reversible cleavage of the Cβ-Cγ bond of L-tyrosine. Here, we present X-ray structures for two enzymatic states that form just before and after the cleavage of the carbon-carbon bond. As for most other pyridoxal 5'-phosphate-dependent enzymes, the first state, a quinonoid intermediate, is central for the catalysis. We captured this relatively unstable intermediate in the crystalline state by introducing substitutions Y71F or F448H in Citrobacter freundii TPL and briefly soaking crystals of the mutant enzymes with a substrate 3-fluoro-L-tyrosine followed by flash-cooling. The X-ray structures, determined at ~2.0 Å resolution, reveal two quinonoid geometries: "relaxed" in the open and "tense" in the closed state of the active site. The "tense" state is characterized by changes in enzyme contacts made with the substrate's phenolic moiety, which result in significantly strained conformation at Cβ and Cγ positions. We also captured, at 2.25 Å resolution, the X-ray structure for the state just after the substrate's Cβ-Cγ bond cleavage by preparing the ternary complex between TPL, alanine quinonoid and pyridine N-oxide, which mimics the α-aminoacrylate intermediate with bound phenol. In this state, the enzyme-ligand contacts remain almost exactly the same as in the "tense" quinonoid, indicating that the strain induced by the closure of the active site facilitates elimination of phenol. Taken together, structural observations demonstrate that the enzyme serves not only to stabilize the transition state but also to destabilize the ground state.


Simultaneous determination of nineteen hallucinogenic tryptamines/beta-calbolines and phenethylamines using gas chromatography-mass spectrometry and liquid chromatography-electrospray ionisation-mass spectrometry.
To investigate the trend of non-controlled drugs of abuse, simultaneous analytical methods were developed using GC-MS and LC-ESI-MS for 8 tryptamines/beta-carbolines, 6 phenethylamines of typically non-controlled substances in Japan, and, additionally, five legally controlled tryptamines and phenethylamines originally found in fungi or plants. Moreover, the proposed methods were applied to analyses of these drugs in 99 kinds of products (a total number of 123 products purchased at adult shops or via the Internet over the past 2 years in Japan), which potentially advertised psychotropic/psychoactive effects. The samples were extracted with methanol under ultrasonication. After centrifugation, the extracts were filtered prior to injections. GC-MS analysis was performed using a DB-5MS capillary column. Regarding the LC-ESI-MS analysis; the separation of the target drugs was optimized on an ODS column in acetonitrile/MeOH (7:3)-10 mM ammonium formate buffer (pH 3.5)/acetonitrile (95:5) by a linear gradient program and a quantitative analysis was carried out by the monitoring of each [M+H]+ in the positive ion mode of ESI-MS. As a result of the analyses using GC-MS and LC-ESI-MS, 5-MeO-DIPT (the synthetic substance known by the street name "Foxy") was found in 8 out of the 99 kinds of products. Additionally, AMT (from brown powder), DMT (from dried plant), harmine and harmaline (from dried plant) were also found in some of the 99 products. These analytical methods could be useful for the investigation of the distribution of the non-controlled psychotropic tryptamines/beta-carbolines and phenethylamines in the market.


Evaluation of different nucleic acid stains for sensitive double-stranded DNA analysis with capillary electrophoretic separation.
This paper outlines the first use of SYTOX Orange, SYTO 82 and SYTO 25 nucleic acid stains for on-column staining of double-stranded DNA (dsDNA) fragments separated by capillary electrophoresis (CE). Low-viscosity, replaceable poly(vinylpyrrolidone) (PVP) polymer solution was used as the sieving matrix on an uncoated fused-silica capillary. The effects of PVP concentration, electric field strength, and incorporated nucleic acid stain concentrations on separation efficiency were examined for a wide range of DNA fragment sizes. Our study was focused on using nucleic acid stains efficiently excitable at a wavelength of 532 nm. Among the five tested nucleic acid stains, SYTOX Orange stain was shown to have the best sensitivity for dsDNA detection by CE. About a 500-fold lower detection limit was obtained compared to commonly used ethidium bromide and propidium iodide. SYTOX Orange stain also provided a wide linear dynamic range for direct DNA quantitation with on-line CE detection. Use of SYTOX Orange stain can greatly improve the measurement of DNA fragments by CE, which will enable an expanded set of applications in genomics and diagnostics.


Clinic expands adolescents' access to care: Health promotion.
A Minneapolis health center opened a mini-clinic in a local high school to provide adolescents with better access to care, appropriate referrals, and health education.


A High-Throughput, Multi-Cell Phenotype Assay for the Identification of Novel Inhibitors of Chemotaxis/Migration.
Chemotaxis and cell migration are fundamental, universal eukaryotic processes essential for biological functions such as embryogenesis, immunity, cell renewal, and wound healing, as well as for pathogenesis of many diseases including cancer metastasis and chronic inflammation. To identify novel chemotaxis inhibitors as probes for mechanistic studies and leads for development of new therapeutics, we developed a unique, unbiased phenotypic chemotaxis-dependent Dictyostelium aggregation assay for high-throughput screening using rapid, laser-scanning cytometry. Under defined conditions, individual Dictyostelium secrete chemoattractants, migrate, and aggregate. Chemotaxis is quantified by laser-scanning cytometry with a GFP marker expressed only in cells after chemotaxis/multi-cell aggregation. We applied the assay to screen 1,280 known compounds in a 1536-well plate format and identified two chemotaxis inhibitors. The chemotaxis inhibitory activities of both compounds were confirmed in both Dictyostelium and in human neutrophils in a directed EZ-TAXIscan chemotaxis assay. The compounds were also shown to inhibit migration of two human cancer cell lines in monolayer scratch assays. This test screen demonstrated that the miniaturized assay is extremely suited for high-throughput screening of very large libraries of small molecules to identify novel classes of chemotaxis/migratory inhibitors for drug development and research tools for targeting chemotactic pathways universal to humans and other systems.


The repair and signaling responses to DNA double-strand breaks.
A DNA double-strand break (DSB) has long been recognized as a severe cellular lesion, potentially representing an initiating event for carcinogenesis or cell death. The evolution of DSB repair pathways as well as additional processes, such as cell cycle checkpoint arrest, to minimize the cellular impact of DSB formation was, therefore, not surprising. However, the depth and complexity of the DNA damage responses being revealed by current studies were unexpected. Perhaps the most surprising finding to emerge is the dramatic changes to chromatin architecture that arise in the DSB vicinity. In this review, we overview the cellular response to DSBs focusing on DNA repair pathways and the interface between them. We consider additional events which impact upon these DSB repair pathways, including regulated arrest of cell cycle progression and chromatin architecture alterations. Finally, we discuss the impact of defects in these processes to human disease.


Advances in Understanding CREB Signaling-Mediated Regulation of the Pathogenesis and Progression of Epilepsy.
Recent studies have indicated that the transcription factor cyclic adenosine monophosphate response element binding protein (CREB) is involved in the etiology of epilepsy. With regard to its transcriptional regulation, CREB phosphorylation is critical for the transmission of multiple extracellular signals, which implicates the activation of downstream target genes in the pathogenesis and progression of epilepsy. This review mainly focuses on recent discoveries of associations between the molecular and structural characteristics of CREB as well as the related CREB signaling pathway and epilepsy.


Innovative production of fungal pulp from Trametes versicolor and its application in a fungal paper box containing clove oil.
The aims of this study were to develop fungal pulp from Trametes versicolor (white-rot fungi) and apply it with clove oil in a paper box to inhibit mold growth on the surface of peanuts. Broken rice media with different sugar solutions (2-10% w w-1) were prepared and then inoculated with T. versicolor mycelium at amounts from 0.5 to 1.5% w w-1. Fungal pulp and commercial paper (50 g) at different ratios (100:0, 70:30, 50:50, 30:70 and 0:100) were mixed and prepared before being placed into a stainless box (5 cm long by 5 cm wide). For the antimicrobial activity against Aspergillus flavus on peanuts, a paper box was incorporated with 2.5, 5 and 7.5% w w-1 of clove oil, eugenol, caryophyllene, and a combination of eugenol and caryophyllene at ratios of 7:1, 4:4, and 1:7. Results indicated that the highest fungal pulp biomass of T. versicolor in broken rice media was found when using 6% sugar with 1% mycelium inoculums. Fungal pulp and commercial paper at the ratio of 70:30 produced the highest value of hardness. The paper box containing clove oil at 7.5% w w-1 inhibited A. flavus on peanuts for at least 28 days while the control had mold growth within 3 days. Combining eugenol and caryophyllene, the main components of clove oil, at the ratio of 7:1 (7.5% w w-1) in the paper box should be a key factor to inhibit A. flavus during storage.


New bacteriological patterns in primary infected aorto-iliac aneurysms: a single-centre experience.
To assess causative pathogens and surgical outcomes in patients with primary infected aorto-iliac aneurysms at our institution. Retrospective study of patients treated at a university hospital between 1992 and 2009. We identified 26 patients (median age, 63 years) with primary infected aneurysms on the aorta (descending thoracic, n = 2; thoraco-abdominal, n = 3; suprarenal, n = 2; infrarenal, n = 15) or iliac arteries (n = 4). Among them, 22 were symptomatic, including 13 with ruptured aneurysms. The causative organisms, identified in 25/26 patients, were Campylobacter fetus, n = 6; Streptococcus pneumoniae, n = 4; Listeria, n = 3; Salmonella, n = 2; Mycobacterium tuberculosis, n = 2; Staphylococcus aureus, n = 1; and other, n = 7. Immune suppression was a feature in 10 (38.4%) patients. Revascularisation was performed in situ in 23 patients (10 allografts, eight grafts, three superficial femoral veins, and 2 stentgrafts) and by extra-anatomic bypass in three patients. Hospital mortality was 23% (in situ group, 17.4%; extra-anatomic group, 66.7%; χ(2)(Yates), P = 0.24). During follow-up in the 20 survivors (median, 48.5 months), there were two non-infection-related deaths (five and 24 months) and six (30%) vascular complications. The bacteriological spectrum of primary infected aorto-iliac aneurysms was wider than previously reported. The availability of new diagnostic tests and increased prevalence of immunosuppression may explain this finding.


Influence of thyroid hormones and methylthiouracil upon elastin content in the liver of rats with carbon tetrachloride-induced hepatic fibrosis.
The effect of administration of thyroid extract and methylthiouracil upon the elastin content of the liver was investigated in healthy and CCl4-poisoned rats. Hyperthyroidism induced a decrease in liver elastin content and hypothyroidism led to an increase of liver elastin. Similar changes were found in rats with carbon tetrachloride-induced hepatic fibrosis. The possible mechanisms of these changes are discussed.


In vivo onset and duration of action varies for botulinum neurotoxin A subtypes 1-5.
To date, over 40 subtypes of botulinum neurotoxins (BoNTs) have been identified. BoNTs are classified into 7 serotypes distinguished primarily by their antigenic properties, but also characterized by their unique SNARE targets and cleavage sites, host specificity, and duration of action. Sequencing efforts in the last decade have identified several subtypes within the serotypes. Subtypes are currently defined as distinct based solely on amino acid sequence comparison, with a similarity cut-off of 2.5% difference. Ten subtypes have been identified for BoNT/A, which is the serotype associated with the most severe human botulism and also the most commonly used serotype for clinical purposes. Analyses of several of these subtypes have revealed distinct characteristics, ranging from differences in cell entry and enzyme kinetics to differences in potency in mice and cell-model specific potency. A long-term activity study in cultured primary neurons has indicated that BoNT/A1, 2, 4, and 5 have a similar duration of action, whereas BoNT/A3 has a significantly shorter duration of action. This report describes an in vivo mouse study, showing that after local injection BoNT/A2 resulted in faster onset of local paralysis than BoNT/A1, 3, 4, and 5, whereas BoNT/A3 resulted in significantly faster recovery of motor-neuron deficiency.


Diagnosis and Surgical Management of Male Pelvic, Inguinal, and Testicular Pain.
Pain occurs in the male genitourinary organs as for any organ system in response to traumatic, infectious, or irritative stimuli. A knowledge and understanding of chronic genitourinary pain can be of great utility to practicing nonurologists. This article provides insight into the medical and surgical management of subacute and chronic pelvic, inguinal, and scrotal pain. The pathophysiology, diagnosis, and medical and surgical treatment options of each are discussed.


Value of Computerized 3D Shape Analysis in Differentiating Encapsulated from Invasive Thymomas.
To retrospectively investigate the added value of quantitative 3D shape analysis in differentiating encapsulated from invasive thymomas. From February 2002 to October 2013, 53 patients (25 men and 28 women; mean age, 53.94 ± 13.13 years) with 53 pathologically-confirmed thymomas underwent preoperative chest CT scans (slice thicknesses ≤ 2.5 mm). Twenty-three tumors were encapsulated thymomas and 30 were invasive thymomas. Their clinical and CT characteristics were evaluated. In addition, each thymoma was manually-segmented from surrounding structures, and their 3D shape features were assessed using an in-house developed software program. To evaluate the added value of 3D shape features in differentiating encapsulated from invasive thymomas, logistic regression analysis and receiver-operating characteristics curve (ROC) analysis were performed. Significant differences were observed between encapsulated and invasive thymomas, in terms of cystic changes (p=0.004), sphericity (p=0.016), and discrete compactness (p=0.001). Subsequent binary logistic regression analysis revealed that absence of cystic change (adjusted odds ratio (OR) = 6.636; p=0.015) and higher discrete compactness (OR = 77.775; p=0.012) were significant differentiators of encapsulated from invasive thymomas. ROC analyses revealed that the addition of 3D shape analysis to clinical and CT features (AUC, 0.955; 95% CI, 0.935-0.975) provided significantly higher performance in differentiating encapsulated from invasive thymomas than clinical and CT features (AUC, 0.666; 95% CI, 0.626-0.707) (p<0.001). Addition of 3D shape analysis, particularly discrete compactness, can improve differentiation of encapsulated thymomas from invasive thymomas.


Identification of exercise-induced ischemia using QRS slopes.
In this work we studied a computer-aided approach using QRS slopes as unconventional ECG features to identify the exercise-induced ischemia during exercise stress testing and demonstrated that the performance is comparable to the experts' manual analysis using standard criteria involving ST-segment depression. We evaluated the performance of our algorithm using a database including 927 patients undergoing exercise stress tests and simultaneously collecting the ECG recordings and SPECT results. High resolution 12-lead ECG recordings were collected continuously throughout the rest, exercise, and recovery phases. Patients in the database were classified into three categories of moderate/severe ischemia, mild ischemia, and normal according to the differences in sum of the individual segment scores for the rest and stress SPECT images. Philips DXL 16-lead diagnostic algorithm was run on all 10-s segments of 12-lead ECG recordings for each patient to acquire the representative beats, ECG fiducial points from the representative beats, and other ECG parameters. The QRS slopes were extracted for each lead from the averaged representative beats and the leads with highest classification power were selected. We employed linear discriminant analysis and measured the performance using 10-fold cross-validation. Comparable performance of this method to the conventional ST-segment analysis exhibits the classification power of QRS slopes as unconventional ECG parameters contributing to improved identification of exercise-induced ischemia.


Plasma levels of nortriptyline and 10-hydroxynortriptyline and treatment-related electrocardiographic changes in the elderly depressed.
Thirty-one elderly depressed patients were treated for seven weeks with nortriptyline with plasma levels kept between 50-180 ng/ml. Electrocardiograms were taken at the third and seventh weeks of treatment. There were significant increases in the PR interval, QTc interval, and heart rate from before and after treatment. However, there were no consistent correlations between electrocardiographic changes during treatment and plasma levels of nortriptyline, 10-hydroxynortriptyline and either of its two isomers (E-10-hydroxynortriptyline, Z-10-hydroxynortriptyline). Increased QRS duration after seven weeks of treatment was correlated with daily dose of nortriptyline.


Antiestrogen action and growth factor regulation.
The basis of the anti-proliferative action of antiestrogens is generally considered to be their ability to inhibit estrogen induced growth pathways by competitively inhibiting the binding of estrogen to the estrogen receptor. Recent data suggest that this may not be the entire story. Moreover, the cascade of events responsible for inhibition of mitogenesis after an initial interaction with the estrogen receptor is poorly understood. Multiple growth factor pathways operate in both normal and neoplastic estrogen/antiestrogen target tissues. While it is unlikely that any single pathway is pivotal, interactions of estrogen and/or antiestrogens with some of these pathways have been implicated in their proliferative effects. The exact molecular mechanisms remain unclear but autocrine, paracrine/juxtacrine, intracrine, and endocrine mediators or various combinations of them are likely to be involved in vivo. Super-imposed on this is the possibility that 'cross-talk' between intracellular signaling pathways may also be involved. Elucidation of such molecular mechanisms will be important with respect to design of novel antiestrogenic/antimitogenic drugs and alternative treatment strategies for both breast and uterine cancer.


Maggot debridement therapy with a direct dressing can cause compression injuries in patients with chronic limb ischemia.
While there are no reports regarding dressing-associated iatrogenic skin ulcer as an adverse event of maggot debridement therapy (MDT), MDT is clinically used on patients with critical limb ischaemia with dermal fragility. Herein we report causes and counter measures for a case of iatrogenic skin ulcer induced by MDT dressing.


Rapid Roll Out of a Pacemaker Home Monitoring Programme: A Patient Perspective.
To assess the safety, efficacy, and patient acceptability of a pacemaker home monitoring (HM) service. All patients receiving a new Biotronik (Biotronik, Berlin, Germany) pacemaker between March 2020 and February 2021 were contacted for participation. Participants were surveyed on their experience of pacemaker HM. HM alerts and remote wound monitoring rates were also assessed. Of the patients contacted, 77% responded, with a mean age of 80.6±9.9 years. Of these, 95.8% agreed that the home monitoring (HM) has been beneficial. Two thirds preferred HM to face-to-face follow-up and two thirds felt safe with HM. Three themes were identified from the comments: reassurance, technology and data security. Forty-one percent (41%) of respondents would like more reassurance that their HM is working, 18% mentioned technology with mixed responses, and 4.7% cited cybersecurity or the use of their personal data as a concern. The average one-way patient journey saved was 24.3±16.7 km (15.1±10.4 miles). One in three HM alerts required action but only 3.4% were urgent. Remote wound review was successful in 59%. The majority of patients prefer HM and almost all think it has been beneficial. It saves significant travel time and provides actionable alerts. The patient experience could be improved by reassuring patients that their device is being monitored.


Potential risk factors for new onset of back pain disability in Japanese workers: findings from the Japan epidemiological research of occupation-related back pain study.
Two-year, prospective cohort data from the Japan epidemiological research of occupation-related back pain study were used for this analysis. To examine the association between a new onset of low back pain (LBP) with disability and potential risk factors among initially symptom-free Japanese workers. Despite strong evidence that psychosocial issues may influence LBP onset among symptom-free persons, these and other LBP risk factors have not been well investigated in the Japanese workplace. Of 5310 participants responding to a self-administered baseline questionnaire (response rate: 86.5%), 3194 (60.2%) completed both 1- and 2-year follow-up questionnaires. The baseline questionnaire assessed individual characteristics, ergonomic work demands, and work-related psychosocial factors. The outcome of interest was new-onset LBP with disability during the follow-up period. Incidence was calculated for the participants who reported no LBP during the past year at baseline. Logistic regression was used to explore risk factors associated with new-onset LBP with disability. Of 836 participants who were symptom-free during the preceding year, 33 (3.9%) reported LBP with disability during the 2-year follow-up. In univariate analyses, "history of LBP," "frequent lifting," "interpersonal stress at workplace," and "monotonous tasks" were all significant predictors of LBP incidence. All of these factors remained statistically significant or almost significant in the multivariate analysis adjusting for the other variables as well as age and sex: adjusted odds ratio (OR) and 95% confidence interval (95% CI) for history of LBP (OR: 3.25, 95% CI: 1.53-6.91), frequent lifting (OR: 3.77, 95% CI: 1.16-12.3), interpersonal stress at workplace (OR: 2.42, 95% CI: 1.08-5.43), and monotonous tasks (OR: 2.21, 95% CI: 0.99-4.94). Both ergonomic and work-related psychosocial factors may predict the development of LBP with disability among previously asymptomatic Japanese workers. Thus, workplace interventions aimed at reducing the incidence of LBP should focus on both ergonomic and psychosocial stress.


The ratio of energy expenditure to nitrogen loss in diverse patient groups--a systematic review.
The ratio of energy expenditure to nitrogen loss respectively of energy to nitrogen provision (E/N) is considered a valuable tool in the creation of an enteral or parenteral formulation. Specific E/N ratios for parenteral nutrition (PN) have not yet been clearly defined. To determine the range of energy expenditure, nitrogen (protein) losses, and E/N ratios for various patient groups, we performed a systematic review of the literature. Medline 1950-2011 was searched for all studies on patients or healthy controls reporting energy expenditure and nitrogen loss at the same time. We identified 53 studies with 91 cohorts which comprised 1107 subjects. Mean TEE ± standard deviation (SD) was 31.2 ± 7.2 kcal/kg BW/day in patients (n = 881) and 35.6 ± 4.3 kcal/kg BW/day in healthy controls (n = 266). Mean total protein loss (TPL) was 1.50 ± 0.57 g/kg BW/day in patients and 0.94 ± 0.24 g/kg BW/day in healthy controls. A non-linear significant correlation was found between TPL and the E/N ratio. The E/N ratio is not a constant value but decreases continuously with increasing protein loss. These variations should be considered in the nutritional support of patients.


Insufficiency fractures of both femoral necks in a young adult caused by osteoporosis: a case report.
Insufficiency fractures of femoral necks due to osteoporosis of unknown aetiology occurring coincidentally on both sides in young adults, have not been described in English literature so far. A 18-year-old young girl presented with a 3-month history of gradual onset of bilateral thigh pain and progressive inability to walk. At the first look, plain radiographs of both hips showed no gross abnormal findings other than some tips on reduced bone mineral density of the femoral necks. MRI was ordered upon a high suspicion of fracture and T1-weighed MRI showed a linear low signal on both femoral necks. Dual-energy X-ray absorptiometry of the lumbar spine and femoral neck showed low mineral density. She was treated surgically with in-situ internal fixation using cannulated screws. The bilateral thigh pain was resolved and walking started immediately after the treatment. Insufficiency fracture of the femoral neck due to osteoporosis should be kept in mind even in young adults complaining of atypical pain without evidence of unusual activities. We used both the terms of stress fracture and insufficiency (osteoporotic) fracture interchangeably as the mechanism of fractures resemble each other.


CEREBO®: A Portable Device for Non-invasive Detection of Intracranial Hematomas in Real Time.
Brain injury has become a silent epidemic and has very low survival and recovery rates because of inaccurate triaging, especially in absence of symptoms. Therefore, a clinical assessment tool for quick onsite detection of intracranial hematoma is necessary. This study aims to assess the efficacy of the near infrared-based device CEREBO® for the non-invasive detection of intracranial hematomas in traumatic head injury patients. Observational, prospective, cohort, single-center study. Forty-four patients recruited from the Department of Neurosurgery of Civil Hospital, Ahmedabad, between 3 and 85 years from June 2018 to March 2020 were examined by CEREBO® and computed tomography (CT) scan within 72 h post-injury or first onset of symptoms to measure the desired parameters. SAS 9.4. The device exhibited high sensitivity (94.87%) and specificity (76.19%) for unilateral hematomas with a positive predictive value (PPV) of 93.67% and a negative predictive value (NPV) of 80%. For bilateral hematomas, the device exhibited a sensitivity of 80%, specificity of 77.78%, PPV 83.33%, and NPV 73.68%. This study establishes the efficacy of CEREBO® to be used as a point-of-care medical screening device for detecting brain hematomas in patients who have had a head injury and is therefore recommended as an adjunct to CT scan. In the triaging or diagnosis phase, it allows for early treatment and thus helps to reduce the secondary injury resulting from the existing and delayed hematomas.


Ubiquitous occurrence of sulfonamides in tropical Asian waters.
Seven sulfonamides, trimethoprim, five macrolides, lincomycin and three tetracyclines were measured in 150 water samples of sewage, livestock and aquaculture wastewater, and river and coastal waters, in five tropical Asian countries. The sum of the concentrations of the target antibiotics in sewage and heavily sewage-impacted waters were at sub- to low-ppb levels. The most abundant antibiotic was sulfamethoxazole (SMX), followed by lincomycin and sulfathiazole. The average concentration of SMX in sewage or heavily sewage-impacted waters was 1720 ng/L in Vietnam (Hanoi, Ho Chi Minh, Can Tho; n=15), 802ng/L in the Philippines (Manila; n=4), 538 ng/L in India (Kolkata; n=4), 282 ng/L in Indonesia (Jakarta; n=10), and 76 ng/L in Malaysia (Kuala Lumpur; n=6). These concentrations were higher than those in Japan, China, Europe, the US and Canada. A predominance of sulfonamides, especially SMX, is notable in these tropical countries. The higher average concentrations, and the predominance of SMX, can be ascribed to the lower cost of the antibiotics. Both the concentration and composition of antibiotics in livestock and aquaculture wastewater varied widely. In many cases, sulfamethazine (SMT), oxytetracycline (OTC), lincomycin, and SMX were predominant in livestock and aquaculture wastewater. Both human and animal antibiotics were widely distributed in the respective receiving waters (i.e., the Mekong River and Manila Bay). SMT/SMX ratios indicate a significant contribution from livestock wastewater to the Mekong River and nearby canals, with an estimated ~10% of river water SMX derived from such wastewater. Mass flow calculations estimate that 12 tons of SMX is discharged annually from the Mekong River into the South China Sea. Riverine inputs of antibiotics may significantly increase the concentration of such antibiotics in the coastal waters.


Dietary habits of patients with regurgitation after colon interposition.
The dietary habits after colon interposition following oesophagectomy in patients without symptoms (n = 8), with regurgitation (n = 22) and in sex- and age-matched healthy controls (n = 20) were studied by a 7-day diary method. The patients ate smaller meals (1080 +/- 90 kJ versus 1810 +/- 151 for the controls, P less than 0.01), more frequently during the day (eight versus five in the controls). Solid and sour foods were preferred, especially by those with regurgitation, compared with controls. The asymptomatic patients consumed more milk and coffee and less cheese, sour milk, meat, fish, eggs, tea and orange juice than the patients with regurgitation. Vegetable fats and medium chain triglycerides were consumed in negligible amounts. The patients with regurgitation had more fluids separately from meals than the asymptomatic patients. The results suggest that intake of vitamin supplements and replacement of animal fats by vegetable fats may be useful in these patients. Much of the dietary difficulties after colon interposition might be the result of the intra-abdominal colon graft loop anastomosed to the antrum; a short graft with a more proximal anastomosis to the lesser curvature merits study in this context.


Long-term stability after high-pull headgear combined with fixed appliances: A retrospective cohort study of growing class II patients.
High-pull headgear with fixed appliances is a common therapeutic plan in class II growing individuals, especially in those with a tendency to hyperdivergence. The stability of this approach has not been adequately assessed in the long-term. The aim of this retrospective study was to assess that long-term stability by means of lateral cephalograms. Seventy-four consecutive patients were included and examined at three time-points: pre-treatment (T1), end of treatment (T2), and at least five years post-treatment (T3). The average initial age of the sample was 9.3 years (standard deviation, SD: 1.6). At T1, the mean ANB was 5.1 degrees (SD: 1.6), the mean SN-PP 5.6 (SD: 3.0) and the mean MP-PP 28.7 degrees (SD: 4.0). The median follow-up time was 8.6 years with an interquartile range of 2.7. A statistically significant, but of small magnitude increase in the SNA angle was noted at T3 compared to T2 after adjusting for the pre-treatment SNA value (mean difference (MD): 0.75; 95% CI: 0.34, 1.15; P<0.001). The inclination of the palatal plane appeared stable in the post-treatment period, while the MP-PP angle showed scarce evidence of decrease during the post-treatment period after adjusting for sex, pre-treatment SNA and SN-PP angles (MD: -2.29; 95% CI: -2.85, -1.74; P<0.001). The sagittal position of the maxilla and the inclination of the palatal plane appeared to be stable after treatment with high-pull headgear and fixed appliances in the long-term. Continuous mandibular growth, both sagittaly and vertically, contributed to the stability of class II correction.


Optimal time for postoperative irradiation of pterygia.
A prospective study of the optimal time for postoperative beta irradiation of pterygia in 54 eyes of 54 patients suggested that immediate postoperative irradiation, given as a single dose of 2000 rad, was associated with a lower rate of recurrence than a similar dose given 4 days postoperatively. The authors suggest that patients with recurrent pterygia, selected to have postoperative beta radiation, receive the radiation as a single dose of 2000 rad at the end of the surgical procedure.


Fatty acid composition of serum lipids in fasting ponies.
Alterations in the fatty acid distribution of total lipid extracts and 4 of the major lipid subclasses of serum in ponies fasted overnight and for 4 and 7 days were determined. Although increases in 16:0, 16:1, and 18:3 omega 3 were observed, decreased amounts of 18:0 and 18:2 omega 6 combined to cause no significant change in the saturated to unsaturated fatty acid ratio in the total extracts. Phospholipid became somewhat preferentially enriched in saturated fatty acids due to a decrease in 18:1, although this response was variable. The free fatty acid and triglyceride fractions both showed increases in relative amounts of 18:3 omega 3 and a decrease in 18:0 and a concomitant change in the saturated to unsaturated fatty acid ratio. This endogenous alteration was most likely due to the mobilization of an increased proportion of polyunsaturated fatty acids from tissue sites with their subsequent incorporation into triglyceride by the liver. It probably reflects the type of forage diet on which the animals had been maintained prior to the study. The fatty acid composition of the cholesteryl ester fractions was unchanged during fasting but contained appreciable amounts of the 18:2 omega 6 fatty acid.


Na+,K(+)-ATPase activity in CNS and noradrenergic neurotransmission: time course of differential desipramine (DMI) effects.
ATPase activities in CNS membranes were studied after administration of desipramine (DMI), a noradrenaline (NA) uptake inhibitor. In a previous paper we reported that Na+,K(+)-ATPase activity significantly increased 3 h after DMI administration (10 mg/kg) in hypothalamus and mesencephalus but not in cerebral cortex and pons-medulla oblongata membranes (Viola et al., Cell. molec. Neurobiol. 1989, 9, 263-271). Here it was observed that Na+,K(+)-ATPase increase induced by acute DMI disappeared at 24 h in hypothalamus but remained during 21 days in mesencephalus. Na+,K(+)-ATPase increase by acute DMI was inhibited when endogenous NA was depleted by the noradrenergic neurotoxin DSP-4 or the NA synthesis inhibitor alpha-methyl-p-tyrosine. On the whole, Mg(2+)-ATPase activity was not modified by treatment. 5'-nucleotidase, another membrane-bound enzyme, was unchanged by acute DMI. The addition of DMI in vitro (50 ng/mg tissue) during Na+,K(+)-ATPase assay failed to affect ATPase activities. Acute DMI effects on Na+,K(+)-ATPase are thus attributable to noradrenergic neurotransmission rather than to non-specific drug-CNS membrane interaction. Furthermore, DMI produces differential effects on membrane Na+,K(+)-ATPase, depending on treatment conditions and CNS area studied.


Successful outcome with eculizumab treatment in a patient with antiphospholipid syndrome presenting with an unusual thrombotic storm.
Catastrophic Antiphospholipid Syndrome (CAPS) is a life-threatening complication of APS requiring complex management to optimize patient outcome. We describe a 54-year-old man with APS with history of splanchnic vein thrombosis, a Factor II G20210A heterozygote, autoimmune hemolytic anemia and thrombocytopenia. He developed sudden onset of severe flank pain due to spontaneous bilateral adrenal hemorrhage while on warfarin with a therapeutic INR. Despite unfractionated heparin and initial clinical improvement, severe thrombocytopenia developed requiring dexamethasone, rituximab, and romiplostim. Hospitalization was complicated further by thrombosis of the inferior vena cava, pulmonary embolism, and painful violaceous patches on his neck and ear cartilages. Punch biopsy of lesions revealed C5b-C9 deposition of small vessel thromboses. Although the inciting event for his thrombotic storm remains uncertain, anti-complement therapy with eculizumab provided rapid and durable lesion resolution. Eculizumab was discontinued after 6 months and patient remains in remission without recurrent thrombosis. This case provides insight on the management of CAPS, including the use of eculizumab.


Current status and approaches to developing press-coated chronodelivery drug systems.
The past several decades have seen the development of many controlled-release preparations featuring constant release rates to maintain drug concentrations in the human body, regardless of the patient's physiological condition. However, long-term constant drug concentrations in the blood and tissue can cause problems such as resistance, tolerability, and drug side effects. People vary considerably in their physiological and biochemical conditions during any 24 h period, due to the circadian rhythm, and thus, the constant delivery of a drug into the body seems both unnecessary and undesirable. If the drug release profile mimics a living system's pulsatile hormone secretion, then it may improve drug efficacy, and reduce the toxicity of a specific drug administration schedule. Medication and treatments provided according to the body's circadian rhythms will result in better outcomes. This may be provided by a chronopharmaceutical dosage regimen with pulsatile release that matches the circadian rhythm resulting from a disease state, so optimizing the therapeutic effect while minimizing side effects. The press coating technique is a simple and unique technology used to provide tablets with a programmable lag phase, followed by a fast, or rate-controlled, drug release after administration. The technique offers many advantages, and no special coating solvent or coating equipment is required for manufacturing this type of tablet. The present review article introduces chronopharmaceutical press-coated products from a patient physiological needs perspective. The contents of this article include biological rhythms and pulsatile hormone secretion in humans, the reasons for using pulsatile drug delivery for disease treatment, recent chronopharmaceutical preparations appearing on the market, updated compilation of all research articles and press-coated delivery techniques, factors affecting the performance and drug release characteristics of press-coated delivery systems, and recent challenges for the press coating technique. We also provide a brief overview of press-coating approaches intended for chronotherapy.


An overview of microplastics characterization by thermal analysis.
Microplastics may be present in the environment as primary microplastics (manufactured) or secondary microplastics (result of the continuous degradation of larger plastic pieces into smaller fragments due to environmental, physicochemical and biotic factors). To fully understand the dynamics of microplastic particles and their environmental effects, harmonized, automated, cheap, rapid and reliable methodologies for sampling, extraction and characterization of microplastic need to be developed. This review focuses on the potential of thermal analytical techniques for microplastics characterization and highlights some of the new trends in this area.


Death effector domain-containing proteins DEDD and FLAME-3 form nuclear complexes with the TFIIIC102 subunit of human transcription factor IIIC.
Death effector domain-containing proteins are involved in important cellular processes such as death-receptor induced apoptosis, NF-kappaB activation and ERK activation. Here we report the identification of a novel nuclear DED-containing protein, FLAME-3. FLAME-3 shares significant sequence (46.6% identical) and structural homology to another DED-containing protein, DEDD. FLAME-3 interacts with DEDD and c-FLIP (FLAME-1) but not with the other DED-containing proteins FADD, caspase-8 or caspase-10. FLAME-3 translocates to, and sequesters c-FLIP in the nucleus upon overexpression in human cell lines. Using the yeast two-hybrid system to identify DEDD-interacting proteins, the TFIIIC102 subunit of human transcription factor TFIIIC was identified as a DEDD- and FLAME-3-specific interacting protein. Co-expression of either DEDD or FLAME-3 with hTFIIIC102 in MCF-7 cells induces the translocation from the cytoplasm and sequestration of hTFIIIC102 in the nucleus, indicating that DEDD and FLAME-3 form strong heterocomplexes with hTFIIIC102 and might be important regulators of the activity of the hTFIIIC transcriptional complex. Consistent with this, overexpression of DEDD or FLAME-3 in 293 cells inhibited the expression of a luciferase-reporter gene under the control of the NF-kappaB promoter. Our data provide the first direct evidence for the involvement of DED-containing proteins in the regulation of components of the general transcription machinery in the nucleus.


Biocompatibility assessment of fibrous nanomaterials in mammalian embryos.
Currently there is a growing interest in the use of nanotechnology in reproductive medicine and reproductive biology. However, their toxic effects on mammalian embryos remain poorly understood. In this work, we evaluate the biocompatibility of two fibrous nanomaterials (NMs): cotton cellulose nanofibers (CNF) and carboxylated multiwalled carbon nanotubes (MWCNT-COOH), by performing an investigation of the embryonic development, gene expression (biomarkers focused on cell stress, apoptosis and totipotency) and in situ apoptosis in bovine embryos. Exposure to NMs did not interfere in preimplantation development or in the incidence of apoptosis in the bovine embryo, but they did affect the gene expression. The results presented are important for an understanding of the toxicity of cotton CNF and MWCNT-COOH on mammalian embryos. To our knowledge, we report the first evaluation of biocompatibility between these NMs on preimplantation embryos, which may open a new window for reproductive biomedical applications.


Acculturation and depressive symptoms in Hispanic older adults: does perceived ethnic density moderate their relationship?
The negative impact of low acculturation on mental health in Hispanic older adults is widely known. The current study examined whether this association varied by a perceived density of people with same racial/ethnic backgrounds in neighborhoods. We hypothesized that the negative impact of low acculturation on mental health would be pronounced when they lived in neighborhoods with a low density of Hispanics. Using data from the Sacramento Area Latino Study on Aging (n = 1,267), hierarchical regression models of depressive symptoms were estimated with sets of predictors: (1) demographic variables, (2) acculturation, (3) perceived density of Hispanics in neighborhoods, and (4) an interaction between acculturation and perceived density of Hispanics. Supporting the hypothesis, the impact of acculturation on depressive symptoms was found to be moderated by the perceived density of Hispanics in neighborhoods. Findings suggest the importance of neighborhood characteristics in the lives of immigrant older adults.


Regulated intracellular degradation of apolipoprotein B in semipermeable HepG2 cells.
A digitonin-permeabilized HepG2 cell system has been developed and used to characterize the pathway responsible for apolipoprotein B100 (apoB) degradation. Degradation of 35S-labeled apoB occurred after an initial lag period and resulted in the loss of approximately 75.8% of apoB after 2 h of chase. The degradation rate in permeabilized cells was slower than that in intact cells, but approximately the same percentage of apoB was lost over 2 h of chase. The loss of intact apoB in permeabilized cells coincided with the appearance of a number of degradation fragments, including 335- and 70-kDa fragments. The detection of a 70-kDa fragment was a sensitive indicator of degradation. ApoB degradation was inhibited at temperatures below 37 degrees C and maximally activated at 42 degrees C. Degradation was also pH-dependent, with inhibition at pH > or = 7.5. ApoB decay was stimulated by ATP supplementation but not GTP and was inhibited in the presence of energy inhibitors. Degradation was not significantly affected by cycloheximide. However, the stability of the 70-kDa fragment was prolonged with cycloheximide. Preincubation of cells with brefeldin A and nocodazole, as well as monensin, did not diminish the degradation of intact apoB or the appearance of the 70-kDa fragment, suggesting a lack of requirement for intracellular transport from the endoplasmic reticulum. Among the various protease inhibitors tested, degradation was most sensitive to N-acetylleucylleucylnorleucinal (ALLN), which abolished the generation of the 70-kDa fragment in a dose-dependent manner. ALLN-sensitive degradation of apoB was unaffected by the calcium ionophore, A23187. Interestingly, degradation of unglycosylated apoB, detected in tunicamycin-pretreated cells, occurred earlier, resulted in generation of additional fragments, and was largely uninhibited by ALLN. In summary, apoB degradation occurs in permeabilized HepG2 cells by a temperature- and pH-sensitive, pre-Golgi degradation system and is catalyzed by a calcium-independent, ALLN-sensitive protease. Specificity of the initial apoB cleavage may require the proper N-linked glycosylation of the protein in the endoplasmic reticulum.


Accurate interrogation of FCGR3A rs396991 in European and Asian populations using a widely available TaqMan genotyping method.
A polymorphism in the receptor for the Fc region of IgG, Fc γ-receptor IIIa (FcγRIIIa, FCGR3A rs396991), has been inconsistently shown in the literature to have an effect on response to monoclonal antibody therapy in several indications. The rs396991 (T/G) polymorphism leads to an F176V substitution and increased affinity for IgG. This variant has proven difficult to genotype accurately, primarily because of extensive homology between the FCGR3A and FCGR3B genes. We have shown that rs396991 can be genotyped by PCR amplification, followed by direct Sanger sequencing of the product, without coamplification of FCGR3B, and that the rs396991 TaqMan assay (C__25815666_10) agrees with Sanger sequencing results in 100% of European and Asian samples tested, but it has a small error rate in African and American populations. C__25815666_10 is therefore suitable to interrogate rs396991 in studies involving Europeans and Asians; however for other populations, the default genotyping method should be PCR followed by Sanger sequencing.


Helicopter transport of sick neonates: a 14-year population-based study.
The Norwegian Helicopter Emergency Medical Service (HEMS) employs anesthesiologists and paramedics who are not formally trained in neonatology to provide stabilization and transport of sick neonates. We describe neonatal transport by HEMS in central Norway and report the outcome. Retrospective analysis of HEMS missions during the 14-year period 1988-2001. A total of 252 neonates were transported, indicating a prevalence of 0.90 per 100 newborn. Median response time was 42 min, on-scene stabilization time 38 min and transportation time 30 min. Median gestational age (GA) was 38 weeks and 4.8% of the neonates were <1000 g and/or <28 weeks. Main clinical problems were respiratory disease, asphyxia and malformations; 30 neonates (12%) died within 1 year. No deaths were transport-related. Tube or ventilator problems were noted in seven out of 66 transports of ventilated neonates. Other technical problems were few. Regarding ventilation, oxygenation and circulation, the clinical condition of most neonates improved during transport and the median temperature rose from 36.5 degrees C to 37.0 degrees C. Hypoglycemia (<2 mmol l(-1)) was documented in 19 missions after transport; of these, eight received a glucose infusion. Four neonates might have profited from HEMS-delivered surfactant therapy. Helicopter Emergency Medical Service in central Norway provides rapid medical assistance in a wide spectrum of neonatal problems, but more attention should be paid to proper ventilation and prevention of hypothermia and hypoglycemia.


Major histocompatibility complex class I molecules are required for the development of insulitis in non-obese diabetic mice.
An early step in the development of autoimmune diabetes is lymphocyte infiltration into the islets of Langerhans of the pancreas, or insulitis. The infiltrate contains both CD4+ and CD8+ T cells and both are required for progression to diabetes in non-obese diabetic (NOD) mice. It has been thought that the CD4+ lymphocytes are the initiators of the disease, the islet invaders, while CD8+ cells are the effectors, the islet destroyers. We question this interpretation because NOD mice lacking MHC class I molecules, hence CD8+ T cells, do not display even insulitis when expected.


Preeclampsia and androgen receptor gene CAG repeat length: results from both children and women.
We studied whether the CAG (encoding glutamine) repeat length polymorphism in the first exon of the androgen receptor (AR) gene is predictive of preeclampsia. Fifty-nine children born after preeclamptic pregnancy (PRE) and 58 control subjects born after normotensive pregnancy (non-PRE) were genotyped for the CAG repeat length of the AR gene. Secondly, the ARCAG repeat lengths of 133 unrelated preeclamptic women and 112 healthy controls were studied. The mean AR gene CAG lengths were compared between the preeclampsia and the control groups. The mean length of the CAG repeat segment among children was significantly shorter in the PRE group compared with the non-PRE group (p = 0.02). Interestingly, the difference between the PRE and the non-PRE boys was even more significant (p = 0.008). Also the distribution of allele frequencies was different, short repeat lengths being overrepresented in the PRE children. However, there were no significant differences in the mean CAG repeat lengths between the unrelated preeclamptic women and their controls, but the shortest CAG repeat lengths were found only in the preeclamptic women. The AR gene CAG repeat length is not a major determinant in the development of preeclampsia. The association of the shortest CAG repeats with preeclampsia is possible, but a larger study group is needed to confirm this finding.


Cognitive Performance Following Ingestion of Glucose-Fructose Sweeteners That Impart Different Postprandial Glycaemic Responses: A Randomised Control Trial.
We aimed to investigate the isolated effect of glycaemia on cognitive test performance by using beverages sweetened with two different glucose-fructose disaccharides, sucrose and isomaltulose. In a randomised crossover design, 70 healthy adults received a low-glycaemic-index (GI) isomaltulose and sucralose beverage (GI 32) and a high-GI sucrose beverage (GI 65) on two occasions that were separated by two weeks. Following beverage ingestion, declarative memory and immediate word recall were examined at 30, 80 and 130 min. At 140 min, executive function was tested. To confirm that the glycaemic response of the test beverages matched published GI estimates, a subsample (n = 12) of the cognitive testing population (n = 70) underwent glycaemic response testing on different test days. A significantly lower value of mean (95% CI) blood glucose concentration incremental area under the curve (iAUC) was found for isomaltulose, in comparison to the blood glucose concentration iAUC value for sucrose, the difference corresponding to -44 mmol/L∙min (-70, -18), p = 0.003. The mean (95% CI) difference in numbers of correct answers or words recalled between beverages at 30, 80 and 130 min were 0.1 (-0.2, 0.5), -0.3 (-0.8, 0.2) and 0.0 (-0.5, 0.5) for declarative memory, and -0.5 (-1.4, 0.3), 0.4 (-0.4, 1.3) and -0.4 (-1.1, 0.4) for immediate free word recall. At 140 min, the mean difference in the trail-making test between beverages was -0.3 sec (-6.9, 6.3). None of these differences were statistically or clinically significant. In summary, cognitive performance was unaffected by different glycaemic responses to beverages during the postprandial period of 140 min.


Mass-forming xanthogranulomatous cholecystitis masquerading as gallbladder cancer.
Mass-forming xanthogranulomatous cholecystitis (XGC), an uncommon inflammatory pathology of gallbladder (GB), masquerades as gallbladder cancer (GBC) and diagnosis is often a histological surprise. A retrospective analysis of prospectively collected database of patients with GB mass operated between August 2009 and September 2012 was conducted to determine clinical, radiological, and intraoperative findings that might aid in the preoperative diagnosis of mass-forming XGC and ascertain their optimal management strategy. Of the 566 patients with GB mass and suspected GBC, 239 were found to be inoperable on preoperative workup and 129 patients had unresectable disease on staging laparoscopy/laparotomy. Of the 198 with resectable disease, 31 were reported as XGC on final histopathology (Group A), while 167 were GBC (Group B). Of these 31 patients, six with an intraoperative suspicion of benign pathology underwent cholecystectomy with segments IVb and V resection, and frozen section histopathology. Twenty-five underwent radical cholecystectomy, with (n = 10) or without (n = 15) adjacent organ resection. In comparison, anorexia and weight loss were significantly more in Group B (p = 0.001 and <0.001). Intraoperatively, empyema and associated gallstones were more common in Group A (p = 0.011 and <0.001). On computed tomography (CT) of the abdomen, continuous mucosal line enhancement and intramural hypodense bands were significantly more in Group A (p < 0.001 and 0.025). While CT abdomen revealed one or more features suggestive of XGC in 64.5 % (20/31) of patients in Group A, 11(35.5 %) did not have any findings suggestive of XGC on imaging. Mass-forming XGC mimics GBC, making preoperative and intraoperative distinction difficult. While imaging findings can help in suspecting XGC, definitive diagnosis require histopathological examination. Presence of typical radiological findings, however, can help in avoiding extended radical resection in selected cases.


Extracting semantic representations from word co-occurrence statistics: a computational study.
The idea that at least some aspects of word meaning can be induced from patterns of word co-occurrence is becoming increasingly popular. However, there is less agreement about the precise computations involved, and the appropriate tests to distinguish between the various possibilities. It is important that the effect of the relevant design choices and parameter values are understood if psychological models using these methods are to be reliably evaluated and compared. In this article, we present a systematic exploration of the principal computational possibilities for formulating and validating representations of word meanings from word co-occurrence statistics. We find that, once we have identified the best procedures, a very simple approach is surprisingly successful and robust over a range of psychologically relevant evaluation measures.


Retrospective analysis of post-tracheostomy complications.
To elucidate patient, disease, and surgical factors that are significantly associated with 90-day tracheostomy complications, readmissions, and mortality. Retrospective case series with chart review. A single academic tertiary care center between 2011 and 2018. Patients who underwent tracheostomy by any technique for any indication were included. Demographic, disease, and operative details were examined. Multivariable analysis was performed to determine factors associated with 90-day complications, 90-day readmissions, and overall mortality. 697 patients were included. 75% of patients had severe comorbidity (ACE-27 score of 3).1 Patients were intubated for 12 days prior to tracheostomy placement on average. The primary indication was ventilator dependence due to critical illness (85%). 74% were performed open and 26% percutaneous. 10% of patients had a tracheostomy-related complication within 90 days. Complications occurred at a median of post-operative day 11, and hemorrhage was most common (n = 35). 14 patients required immediate return to the operating room, and 3 patients died of their complication, all within 3 days of tracheostomy placement. 40% of patients undergoing tracheostomy died within 30 days. In multivariable analysis, only a documented difficult tracheostomy placement was significantly associated with a 90-day complication. While complications after tracheostomy are infrequent, they are often severe. A heightened level of preparedness to immediately manage accidental tracheostomy decannulation or hemorrhage is required for patients with a difficult tracheostomy placement. 30-day mortality is high, which reinforces the need for multi-disciplinary evaluation, including palliative care, to determine appropriate candidacy for tracheostomy.


Randomized controlled trial of daily teriparatide, weekly high-dose teriparatide, or bisphosphonate in patients with postmenopausal osteoporosis: The TERABIT study.
The effects of daily teriparatide (20 μg) (D-PTH), weekly high-dose teriparatide (56.5 μg) (W-PTH), or bisphosphonates (BPs) on areal bone mineral density (aBMD), bone turnover markers (BTMs), volumetric BMD (vBMD), microarchitecture, and estimated strength were investigated in postmenopausal osteoporosis patients. The study participants were 131 women with a history of fragility fractures. They were randomized to receive D-PTH, W-PTH, or BPs (alendronate or risedronate) for 18 months. Dual-energy X-ray absorptiometry (DXA), BTMs, and high-resolution peripheral quantitative CT (HR-pQCT) parameters were evaluated at baseline and after 6 and 18 months of treatment. The primary endpoint was the change (%) in cortical thickness (Ct.Th) after 18 months' treatment compared with baseline. DXA showed that D-PTH, W-PTH, and BPs increased lumbar spine aBMD (+12.0%, +8.5%, and +6.8%) and total hip aBMD (+3.0%, +2.1%, and +3.0%), but D-PTH and W-PTH decreased 1/3 radius aBMD (-4.1%, -3.0%, -1.4%) after 18 months. On HR-pQCT, D-PTH increased trabecular vBMD (Tb.vBMD) at the distal radius and tibia after 18 months (+6.4%, +3.7%) compared with the BPs group, decreased cortical volumetric tissue mineral density (Ct.vTMD) (-1.8%, -0.9%) compared with the other groups, increased Ct.Th (+1.3%, +3.9%), and increased failure load (FL) (+4.7%, +4.4%). W-PTH increased Tb.vBMD (+5.3%, +1.9%), maintained Ct.vTMD (-0.7%, +0.2%) compared with D-PTH, increased Ct.Th (+0.6%, +3.6%), and increased FL (+4.9%, +4.5%). The BPs increased Tb.vBMD only in the radius (+2.0%, +0.2%), maintained Ct.vTMD (-0.6%, +0.3%), increased Ct.Th (+0.5%, +3.4%), and increased FL (+3.9%, +2.8%). D-PTH and W-PTH comparably increased Ct.Th, the primary endpoint. D-PTH had a strong effect on trabecular bone. Although D-PTH decreased Ct.vTMD, it increased Ct.Th and total bone strength. W-PTH had a moderate effect on trabecular bone, maintained Ct.vTMD, and increased Ct.Th and total bone strength to the same extent as D-PTH.


Social memory, amnesia, and autism: brain oxytocin secretion is regulated by NAD+ metabolites and single nucleotide polymorphisms of CD38.
Previously, we demonstrated that CD38, a transmembrane protein with ADP-ribosyl cyclase activity, plays a critical role in mouse social behavior by regulating the release of oxytocin (OXT), which is essential for mutual recognition. When CD38 was disrupted, social amnesia was observed in Cd38 knockout mice. The autism spectrum disorders (ASDs), characterized by defects in reciprocal social interaction and communication, occur either sporadically or in a familial pattern. However, the etiology of ASDs remains largely unknown. Therefore, the theoretical basis for pharmacological treatments has not been established. Hence, there is a rationale for investigating single nucleotide polymorphisms (SNPs) in the human CD38 gene in ASD subjects. We found several SNPs in this gene. The SNP rs3796863 (C>A) was associated with high-functioning autism (HFA) in American samples from the Autism Gene Resource Exchange. Although this finding was partially confirmed in low-functioning autism subjects in Israel, it has not been replicated in Japanese HFA subjects. The second SNP of interest, rs1800561 (4693C>T), leads to the substitution of an arginine (R) at codon 140 by tryptophan (W; R140W) in CD38. This mutation was found in four probands of ASD and in family members of three pedigrees with variable levels of ASD or ASD traits. The plasma levels of OXT in ASD subjects with the R140W allele were lower than those in ASD subjects lacking this allele. The OXT levels were unchanged in healthy subjects with or without this mutation. One proband with the R140W allele receiving intranasal OXT for approximately 3years showed improvement in areas of social approach, eye contact and communication behaviors, emotion, irritability, and aggression. Five other ASD subjects with mental deficits received nasal OXT for various periods; three subjects showed improved symptoms, while two showed little or no effect. These results suggest that SNPs in CD38 may be possible risk factors for ASD by abrogating OXT function and that some ASD subjects can be treated with OXT in preliminary clinical trials.


Development of a multiplex microsatellite marker set for the study of the solitary red mason bee, Osmia bicornis (Megachilidae).
Solitary bees, such as the red mason bee (Osmia bicornis), provide important ecosystem services including pollination. In the face of global declines of pollinator abundance, such haplodiploid Hymenopterans have a compounded extinction risk due to the potential for limited genetic diversity. In order to assess the genetic diversity of Osmia bicornis populations, we developed microsatellite markers and characterised them in two populations. Microsatellite sequences were mined from the recently published Osmia bicornis genome, which was assembled from DNA extracted from a single male bee originating from the United Kingdom. Sequences were identified that contained dinucleotide, trinucleotide, and tetranucleotide repeat regions. Seventeen polymorphic microsatellite markers were designed and tested, sixteen of which were developed into four multiplex PCR sets to facilitate cheap, fast and efficient genotyping and were characterised in unrelated females from Germany (n = 19) and England (n = 14). The microsatellite markers are highly informative, with a combined exclusion probability of 0.997 (first parent), which will enable studies of genetic structure and diversity to inform conservation efforts in this bee.


Solubility of toluene, benzene and TCE in high-microbial concentration systems.
We report measurements of solubility limits for benzene, toluene, and TCE in systems that contain varying levels of biomass up to 0.13 g mL(-1) for TCE and 0.25 g mL(-1) for benzene and toluene. The solubility limit increased from 21 to 48 mM when biomass (in the form of yeast) was added to aqueous batch systems containing benzene. The toluene solubility limit increased from 4.9 to greater than 20mM. For TCE, the solubility increased from 8mM to more than 1000 mM. Solubility for TCE (trichloroethylene) was most heavily impacted by biomass levels, changing by two orders of magnitude as the microbial concentrations approach those in biofilms.


The amniotic band disruption complex. The pathogenesis of oblique facial clefts; an experimental study in the foetal rat.
This paper reports the findings of an experimental foetal rat study which has shown that it is possible to produce facial clefts by stimulating the tethering and disruptive effects of amniotic bands. Of the 10 experimental foetuses, 9 (90%) survived the in utero surgery and are included in the present study. Facial and palatal clefts were produced in all 9 (100%) of these foetuses.


Proteolysis: a double-edged sword for the development of amyloidoses.
The yeast Saccharomyces cerevisiae has proven to be a useful model system to investigate the mechanism of prion generation and inheritance, to which studies in Sup35 made a great contribution. Recent studies demonstrated that 'protein misfolding and aggregation' (i.e. amyloidogenesis) is a common principle underlying the pathogenesis of neurodegenerative diseases including prion, amyotrophic lateral sclerosis (ALS), Perkinson's (PD), Alzheimer's (AD) diseases and polyglutamine (polyQ) diseases such as spinocerebellar ataxia (SCA) and Hantington's disease (HD). By these findings, the yeast has again been drawing increased attention as a useful system for studying neurodegenerative proteinopathies. So far, it has been reported that proteolytic cleavage of causative amyloidogenic proteins might affect the pathogenesis of the respective neurodegenerative diseases. Although those reports provide a clear phenomenological description, in the majority of cases, it has remained elusive if proteolysis is directly involved in the pathogenesis of the diseases. Recently, we have demonstrated in yeast that proteolysis suppresses prion generation. The yeast-based strategy might make a breakthrough to the unsolved issues.


Immunohistological localization of IgG1, IgA and secretory component in the bovine mammary gland during involution.
Immunoperoxidase methods were used to localize secretory component, immunoglobulin A and immunoglobulin G1 in mammary tissue from dairy cows. In lactating tissue, immunostaining for immunoglobulin A and secretory component was observed primarily in the luminal contents of alveoli. By day 2 of involution, alveolar epithelial cells stained for both immunoglobulin A and secretory component. Staining of alveolar epithelial cells for immunoglobulin A and secretory component continued throughout the period of mammary involution. No staining for secretory component was observed in the interalveolar stromal area. Immunoglobulin G1 immunostaining was localized primarily in the interalveolar areas in lactating tissue, but was localized at the apical and basolateral surface of alveolar cells on day 2 of involution. In contrast to immunoglobulin A, immunoglobulin G1 staining of epithelial cells did not persist and was primarily in the interalveolar areas by day 4. These results suggest that an increased localization of immunoglobulin G1 in bovine mammary epithelial cells may occur transiently in early involution, while an increase in immunoglobulin A and secretory component localization in epithelial cells persists throughout involution.


On the nature of arterial and lung calcifications induced in cattle by Solanum glaucophyllum.
The chemical and crystalline composition of the mineral deposited in ossified arterial and lung tissues of cows affected by enteque seco (enteque ossificans) has been investigated. The arterial calcification precedes that of lung. The latter was observed only in animals with a calcification of the aorta above 150 mumoles Ca per gram of wet tissue. The mineral deposited in arterial and lung tissues appeared to be a poorly crystallized apatite; following incineration the diffraction patterns of carbonate apatite and magnesium whitlockite, respectively, were found.


Relationship between trabecular vertebral body density and fractures: a quantitative definition of spinal osteoporosis.
To evaluate the relationship between vertebral fractures and trabecular vertebral body density (TVBD), as measured by computed tomography (CT), we evaluated 110 female and 38 male patients referred consecutively to our clinic for an osteoporosis evaluation. Number of fractures per patient and TVBD was negatively correlated in both males and females (r = -.64, P less than .001 and r = -.69, P less than .001, respectively). Based on this relationship and that between percent of patients with fracture and TVBD, we devised four different approaches to calculate the fracture threshold. (1) Because the x-axis intercept of this regression line represents the TVBD value at zero fractures, this intercept can be considered the fracture threshold, which was 103 mg/cm3 for females and 132 mg/cm3 for males. (2) Breakpoint analysis of the relationship between the number of vertebral fractures per patient v TVBD gave a fracture threshold value of 98 mg/cm3 for females, but for males we were unable to compute a threshold value because of the small sample size. The percentage of patients with fractures was also negatively correlated with TVBD for males and females (r = -.98, P less than .001, and r = -.94, P less than .001, respectively). (3) the x-axis intercept of this relationship, which represents the fracture threshold, was 123 mg/cm3 for males and 101 mg/cm3 for females. (4) The fracture threshold, calculated as the mean TVBD + 2 SD for patients with fracture(s), was 120 and 92 mg/cm3 for males and females, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)


Isolation and characterization of mitochondria from human B lymphoblastoid cell lines.
Mitochondria were isolated from detergent-treated Epstein-Barr virus-transformed human lymphocytes to examine their potential use in the study of the functional expression of genetic disorders of the respiratory chain. The increase of cytochrome c oxidase activity in the mitochondrial fraction indicated a 6-fold purification of intact mitochondria. Polarographic and spectrophotometric studies revealed that the isolated mitochondria were functionally well preserved. Furthermore, the isolated mitochondria supported an active in organello protein synthesis, which was dependent on the presence of a respiratory substrate generating ATP and was essentially abolished by chloramphenicol or by a specific respiratory chain inhibitor, such as antimycin. Thus, B lymphoblastoid cell lines constitute a valuable source of mitochondria to investigate mitochondrial functions in patients affected by respiratory chain disorders.


Early-juvenile Batten's disease--a recognisable sub-group distinct from other forms of Batten's disease. Analysis of 5 patients.
In most cases where rectal biopsy is performed to diagnose Batten's disease, there is good correlation between biopsy appearance, age of onset, clinical course and electrophysiological parameters. As a result 3 forms of the disease have been recognised; infantile, late infantile and juvenile. In a review of rectal biopsy in Batten's disease at the Hospital for Sick Children, Great Ormond Street, we have studied the few cases in which no such correlation appeared to exist. In 5 the features are sufficiently similar to suggest a further recognisable sub-group which could be descriptively called "early juvenile". The clinical course, electrophysiological features and the absence of vacuolated lymphocytes in this subgroup are as found in the late infantile form, whereas the biopsy findings are identical to those of the juvenile form. By analogy with some of the mucopolysaccharidoses we speculate that the genes of the late infantile and juvenile forms of Batten's disease are allelic and that the "early juvenile" sub-group is a genetic compound presenting as an intermediate phenotype.