Transcorneal and transscleral iontophoresis of dexamethasone phosphate using drug loaded hydrogel.
To evaluate dexamethasone penetration to the eye after a short transcorneal and transscleral iontophoresis using a drug loaded hydrogel assembled on a portable iontophoretic device. Iontophoresis of dexamethasone phosphate was studied in healthy rabbits using drug loaded disposable HEMA hydrogel sponges and portable iontophoretic device. Corneal iontophoretic administration was performed with a current intensity of 1 mA for 1 and 4 min. Transconjunctival and transscleral iontophoresis were performed twice for 2 min at two near places in the pars-plana area, on the conjunctival membrane or directly on the sclera. Dexamethasone concentrations were assayed using HPLC. Dexamethasone levels in the rabbit cornea after a single transcorneal iontophoresis for 1 min were up to 30 fold higher compared to those obtained after frequent eye drop instillation. Also, high drug concentrations were obtained in the retina and sclera 4 h after transscleral iontophoresis. A short low current non-invasive iontophoretic treatment using dexamethasone-loaded hydrogels has potential clinical value in increasing drug penetration to the anterior and posterior segments of the eye.


Site of Metabolism Prediction Based on ab initio Derived Atom Representations.
Machine learning models for site of metabolism (SoM) prediction offer the ability to identify metabolic soft spots in low-molecular-weight drug molecules at low computational cost and enable data-based reactivity prediction. SoM prediction is an atom classification problem. Successful construction of machine learning models requires atom representations that capture the reactivity-determining features of a potential reaction site. We have developed a descriptor scheme that characterizes an atom's steric and electronic environment and its relative location in the molecular structure. The partial charge distributions were obtained from fast quantum mechanical calculations. We successfully trained machine learning classifiers on curated cytochrome P450 metabolism data. The models based on the new atom descriptors showed sustained accuracy for retrospective analyses of metabolism optimization campaigns and lead optimization projects from Bayer Pharmaceuticals. The results obtained demonstrate the practicality of quantum-chemistry-supported machine learning models for hit-to-lead optimization.


Batch and semi-continuous experiments examining the sludge mesophilic anaerobic digestive performance with different varieties of rice straw.
Anaerobic co-digestion of excess sludge (ES) and different varieties of rice straw including indica rice straw (IRS), japonica rice straw (JRS) and glutinous rice straw (GRS) was investigated in batch and semi-continuous experiments. The batch experiment results showed that the GRS addition presents the highest hydrolysis and methanogenesis rates, its cumulative methane yield (CMY) was 305.75 mL/g VS and its average methane content was 60.56%. After digestion, the structure of GRS was almost completely destroyed, which was beneficial to the degradation of lignocellulose. The digestive process is affected by the abundance of Actinobactereria, Proteobacteria, Methanosaetae and Methanosarcina. The results of semi-continuous digestion were similar to batch digestion. In addition, the addition of GRS increased TN concentration in biogas residue and TP concentration in biogas slurry, but was not conducive to the subsequent dehydration of sludge.


Assessment of Corticosteroid Therapy and Death or Disability According to Pretreatment Risk of Death or Bronchopulmonary Dysplasia in Extremely Preterm Infants.
Meta-analyses suggest that corticosteroids may be associated with increased survival without cerebral palsy in infants at high risk of bronchopulmonary dysplasia (BPD) but are associated with adverse neurologic outcomes in low-risk infants. Whether this association exists in contemporary practice is uncertain because most randomized clinical trials administered corticosteroids earlier and at higher doses than currently recommended. To evaluate whether the pretreatment risk of death or grade 2 or 3 BPD at 36 weeks' postmenstrual age modified the association between postnatal corticosteroid therapy and death or disability at 2 years' corrected age in extremely preterm infants. This cohort study analyzed data on 482 matched pairs of infants from 45 participating US hospitals in the National Institute of Child Health and Human Development Neonatal Research Network Generic Database (GDB). Infants were included in the cohort if they were born at less than 27 weeks' gestation between April 1, 2011, and March 31, 2017; survived the first 7 postnatal days; and had 2-year death or developmental follow-up data collected between January 2013 and December 2019. Corticosteroid-treated infants were propensity score matched with untreated controls. Data were analyzed from September 1, 2019, to November 30, 2022. Systemic corticosteroid therapy to prevent BPD that was initiated between day 8 and day 42 after birth. The primary outcome was death or moderate to severe neurodevelopmental impairment at 2 years' corrected age. The secondary outcome was death or moderate to severe cerebral palsy at 2 years' corrected age. A total of 482 matched pairs of infants (mean [SD] gestational age, 24.1 [1.1] weeks]; 270 males [56.0%]) were included from 656 corticosteroid-treated infants and 2796 potential controls. Most treated infants (363 [75.3%]) received dexamethasone. The risk of death or disability associated with corticosteroid therapy was inversely associated with the estimated pretreatment probability of death or grade 2 or 3 BPD. The risk difference for death or neurodevelopmental impairment associated with corticosteroids decreased by 2.7% (95% CI, 1.9%-3.5%) for each 10% increase in the pretreatment risk of death or grade 2 or 3 BPD. This risk transitioned from estimated net harm to benefit when the pretreatment risk of death or grade 2 or 3 BPD exceeded 53% (95% CI, 44%-61%). For death or cerebral palsy, the risk difference decreased by 3.6% (95% CI, 2.9%-4.4%) for each 10% increase in the risk of death or grade 2 or 3 BPD and transitioned from estimated net harm to benefit at a pretreatment risk of 40% (95% CI, 33%-46%). Results of this study suggested that corticosteroids were associated with a reduced risk of death or disability in infants at moderate to high pretreatment risk of death or grade 2 or 3 BPD but with possible harm in infants at lower risk.


Lysis protein encoded by plasmid ColA-CA31. Gene sequence and export.
A gene, cal, coding for a polypeptide needed for the release of colicin A from Escherichia coli cells has been identified by transposon insertion. The cal gene was located on the ColA plasmid map adjacent to cai, the gene coding for colicin A immunity protein, and therefore 592 bases downstream from caa, the structural gene for colicin A. Transcription of cal is in the same direction as caa, that is in the opposite direction to cai. Its sequence has been determined and the predicted amino acid composition features a basic N-terminal end followed by a serie of hydrophobic residues similar to the signal sequence in precursors of exported proteins. The C-terminal part also contains a core of hydrophobic residues. The overall amino acid sequence of the cal protein is homologous to that of lytic proteins encoded by the related plasmids pColE1, and pCloDF13. The cal protein has been identified on urea-SDS-polyacrylamide gels by selective labelling with various radioactive amino acids and its synthesis is co-induced with that of colicin A. The cal protein undergoes slow processing with loss of the N-terminal "signal" region and the mature form is released into the medium together with colicin A.


Enterobacter cloacae SLD1a-1 gains a selective advantage from selenate reduction when growing in nitrate-depleted anaerobic environments.
Enterobacter cloacae SLD1a-1 is capable of the complete reduction of selenate to selenium and the initial reaction is catalysed by a membrane-bound selenate reductase. In the present study, continuous culture experiments were employed to investigate the possibility that selenate reduction, via the selenate reductase, might provide sufficient energy to maintain cell viability when deprived of the preferred anaerobic terminal electron acceptor nitrate. The evidence presented indicates that the selenate reductase supports slow growth that retards the wash-out of the culture when switching to nitrate-depleted selenate-rich medium, and provides a proton motive force for sustained cell maintenance. In contrast, a strain of E. cloacae (sub sp. cloacae) that does not readily reduce selenate, cannot sustain cell maintenance when switching to a selenate-rich medium. This work demonstrates for the first time that respiratory linked selenate reduction gives E. cloacae SLD1a-1 a selective advantage when inhabiting selenate-contaminated environments and highlights the suitability of utilising E. cloacae SLD1a-1 when developing selenium remediation strategies.


Phytochemical and in silico studies for potential constituents from Centaurium spicatum as candidates against the SARS-CoV-2 main protease and RNA-dependent RNA polymerase.
In the present study, a new secoiridoid glycoside lisianthoside II 1, along with seven known compounds 2-8, were isolated from Centaurium spicatum L. In-silico molecular docking and molecular dynamic simulation against SARS-CoV-2 Main protease (Mpro) and RNA-dependent RNA polymerase (RdRp) were conducted. The affinity docking scores revealed that 8 is the best bound ligand to Mpro active site with binding energy of -14.9877 kcal/mol (RSMD = 1.16 Å), while 6 was the highest against RdRp (-16.9572 kcal/mol, RMSD = 1.01 Å). Moreover, the molecular dynamic simulation revealed that 8 with a (ΔG) of -7.9 kcal/mol (RMSD value of 2.6 Å) and 6 (RMSD value of 1.6 Å) and binding free energy (ΔG) of -7.1 kcal/mol achieved the highest stability over 50 ns of MDS inside the Mpro and RdRp enzyme's active site, respectively. Hence, the isolated compounds could be a good lead for development of new leads targeting COVID-19.


The global landscape of drug development of trastuzumab biosimilars.
Limited access to prevention, screening, timely diagnosis, affordable and effective treatment is the main driver of disparities in cancer care. The high costs represent a common barrier for the affordable access to biological agents. The only biological agent included in the World Health Organization list of essential medicines for the management of breast cancer is trastuzumab, and its biosimilars. We reviewed the pivotal trials for approved trastuzumab biosimilars and searched the proposed biosimilars under investigation in clinical trials, then mapped them against metrics of national epidemiology and health system capacity. Thirteen clinical trials were extracted. Across 40 countries and 825 research facilities, more than three- fourth enrolling centers were located in low- and middle- income countries (n = 633) Correlation analysis suggested that the research and drug development of biosimilars was prioritized in settings where weaker health systems can result in poorer cancer- related outcomes and higher out of pocket expenses (range of r: -0,37; + 0.53; all p-values <0.05). Eleven countries had no biosimilars approved at the time of the analysis: in these countries, the drug development of biosimilars was more represented. The drug development of biosimilars occurs significantly in low- and middle- income countries. Our correlative findings must be interpreted cautiously, to understand if the biosimilar drug development is congruent with the national health priorities of cancer control. Health, societal and economic impacts on populations should be accounted, as relevant metrics for clinical trial implementations. The implementation of cancer research should be developed under the national cancer control priorities, to ensure consistency across the evidence- informed relevant national policies, thus result in a population health impact. Biosimilars drug development can drive tangible population health benefits by accelerating the uptake of more sustainable health interventions and shaping more efficient regulatory environments.


Ectopic breast cancer: a case report and review of the Japanese literature.
We report a rare case of ectopic breast cancer in a 68-year-old woman, successfully treated by surgery and chemotherapy. We found 37 cases including our case in the Japanese literature between 1933 to 1997. Our review demonstrated that the clinical features of Japanese patients who developed ectopic breast cancer was similar to previous foreign reports.


Extending half-life in coagulation factors: where do we stand?
The human coagulation factor VIII (FVIII) and von Willebrand factor (VWF) are two distinct glycoproteins that circulate in the plasma as a non-covalently bound complex (VWF/FVIII complex). Deficiencies or structural defects in FVIII and VWF are responsible for the most common inherited plasma bleeding disorders haemophilia A and von Willebrand disease (VWD), respectively. Current therapies for the treatment of haemophilia have favourable efficacy, tolerability and safety profiles. However, multiple, frequent infusions are usually required to manage a bleeding episode, owing to the short half-life of FVIII. This makes treatment inconvenient and impacts patient quality of life. Several strategies are currently being pursued in an attempt to reduce the number of infusions required per bleeding episode. One of the more promising approaches involves prolonging the half-life of FVIII. This article summarizes the methods that are being used to extend FVIII half-life.


Clinical and pathologic manifestation of oesophagostomosis in African great apes: does self-medication in wild apes influence disease progression?
Nodular worms (Oesophagostomum spp.) are common intestinal parasites found in cattle, pig, and primates including humans. In human, they are responsible for serious clinical disease called oesophagostomosis resulting from the formation of granulomas, caseous lesions or abscesses in intestinal walls. In wild great apes, the fecal prevalence of this parasite is high, but little information is available concerning the clinical signs and lesions associated. In the present study, we describe six cases of multinodular oesophagostomosis in free-ranging and ex-captive chimpanzees and captive gorillas caused by Oesophagostomum stephanostomum. While severe clinical signs associated with this infection were observed in great apes raised in sanctuaries, nodules found in wild chimpanzees do not seem to affect their health status. One hypothesis to explain this difference would be that in wild chimpanzees, access to natural environment and behavior such as rough leaves swallowing combined with ingestion of plants having pharmacological properties would prevent severe infection and decrease potential symptoms.


One year in autoimmunity.
The past 12 months have witnessed several critical advancements and new particularly promising lines of evidence on the pathogenesis and clinical management of autoimmune diseases. The aim of the present article is to briefly review such progress in specific conditions as well as in autoimmunity in general by highlighting papers published in the Journal of Autoimmunity and in Autoimmunity Reviews. These data cumulatively support the hypothesis that autoimmunity is multifactorial but that specific diseases cluster together; they also continue to reflect the critical relationships between genetics and environmental triggers.


Anatomical study of the deep branch of the ulnar nerve and application to selective neurectomy in the treatment of spasticity of the first web space.
Spasticity of the first web space is common in upper limb spasticity. Selective neurectomy is one of the treatments that can reduce spasticity. The purpose of this study was to describe the variations of the deep motor branch of the ulnar nerve for the adductor pollicis (AP) and the first dorsal interosseous muscle (DIO) to assess the feasibility of selective neurectomy and suggest an ideal surgical approach. The deep branch of the ulnar nerve (DBUN) was dissected in 21 hands. Measurements included the distance between the point of passage of the DBUN between the two heads of the adductor and three anatomical landmarks: the bi-styloid line, the flexor carpi radialis and the pisiform bone, and the number and mode of divisions of each branch. The point of passage of the DBUN between the two heads of the adductor is very constant respective to the landmarks. The DBUN gives off 1-3 branches each for the oblique head of the AP, the transverse head, and the first DIO. Muscles receive more than one branch in 95% cases for the oblique head and 62% of cases for the transverse head, and 100% for the DIO. This anatomical study suggests that selective neurectomy is feasible for the AP and first DIO muscles in most cases. An ideal approach for selective neurectomy of these muscles should start from the point of passage of the DBUN between the two heads of the AP. This point is easily identified with the help of the described landmarks.


Molecular cloning and expression of a proteinase gene from Lactococcus lactis subsp. cremoris H2 and construction of a new lactococcal vector pFX1.
The 6.5 kb HindIII DNA fragment of the Lactococcus lactis subsp. cremoris H2 plasmid pDI21 was cloned into Escherichia coli POP13 with lambda NM1149, and also directly into Lactococcus lactis subsp. lactis 4125 using a newly-constructed broad host-range vector pFX1. Proteinase was expressed in both transformed organisms. The proteinase resembles a PI type since it preferentially degraded beta-casein. The restriction map of the 6.5 kb proteinase gene fragment has minor differences from those of published plasmid proteinase genes. High-efficiency electroporation with pFX1 provides a direct approach for gene cloning in lactococci.


Detection of arsenic-containing hydrocarbons in a range of commercial fish oils by GC-ICPMS analysis.
The present study describes the use of a simple solid-phase extraction procedure for the extraction of arsenic-containing hydrocarbons from fish oil followed by analysis using gas chromatography (GC) coupled to inductively coupled plasma mass spectrometry (ICPMS). The procedure permitted the analysis of a small sample amount, and the method was applied on a range of different commercial fish oils, including oils of anchovy (Engraulis ringens), Atlantic herring (Clupea harengus), sand eel (Ammodytes marinus), blue whiting (Micromesistius poutassou) and a commercial mixed fish oil (mix of oils of Atlantic herring, Atlantic cod (Gadus morhua) and saithe (Pollachius virens)). Total arsenic concentrations in the fish oils and in the extracts of the fish oils were determined by microwave-assisted acid digestion and ICPMS. The arsenic concentrations in the fish oils ranged from 5.9 to 8.7 mg kg(-1). Three dominant arsenic-containing hydrocarbons in addition to one minor unidentified compound were detected in all the oils using GC-ICPMS. The molecular structures of the arsenic-containing hydrocarbons, dimethylarsinoyl hydrocarbons (C17H38AsO, C19H42AsO, C23H38AsO), were verified using GC coupled to tandem mass spectrometry (MS/MS), and the accurate masses of the compounds were verified using quadrupole time-of-flight mass spectrometry (qTOF-MS). Additionally, total arsenic and the arsenic-containing hydrocarbons were studied in decontaminated and in non-decontaminated fish oils, where a reduced arsenic concentration was seen in the decontaminated fish oils. This provided an insight to how a decontamination procedure originally ascribed for the removal of persistent organic pollutants affects the level of arsenolipids present in fish oils.


Sex Differences in Mate Choice Preference Characteristics of Aequidens rivulatus.
The mating roles of males and females, to a certain extent, are dynamic and variable. Several factors influence the mate choice process. Nonetheless, the main preference features have not yet been fully understood in Aequidens rivulatus. In this study, because of its natural pairing characteristics, A. rivulatus was selected to explore the mate choice preferences of different sexes. Specifically, male and female behavioral performances were described and quantified through a "no-choice paradigm" during mate choice. A total of 12 behavioral performances were defined in male mate choice (experiment 1), whereas 14 behavioral performances were defined in female mate choice (experiment 2). According to the obtained results, unselected females did not display any proactive behaviors in experiment 1, whereas unselected males exhibited proactive behaviors in experiment 2, including quivering, nipping, tail beating, swimming up and down, and aggression. It was also found that both male and female individuals tend to express dislike rather than like. Those behaviors with higher frequencies (e.g., quivering) often mean less energy expenditure, thus easier repeatability. Moreover, principal component analysis (PCA) was employed to extract and identify mate choice preference features. Preliminary results indicated that male preferences for a mate were mainly associated with body size, behavioral intention, and appearance, whereas the intensity of female preferences was in the order of body size, appearance, and behavioral intention. In addition, sex hormone levels were associated with mate choices.


Risk Factors for Flap Loss: Analysis of Donor and Recipient Vessel Morphology in Patients Undergoing Microvascular Head and Neck Reconstructions.
In microvascular head and neck reconstruction, various factors such as diabetes, alcohol consumption, and preoperative radiation hold a risk for flap loss. The primary objective of this study was to examine the vessel morphology of both recipient and donor vessels and to identify predictors for changes in the diameters of H.E.-stained specimens associated with flap loss in a prospective setting. Artery and vein samples (N = 191) were collected from patients (N = 100), with sampling from the recipient vessels in the neck area and the donor vessels prior to anastomosis. External vessel diameter transverse (ED), inner vessel diameter transverse (ID), thickness vessel intima (TI), thickness vessel media (TM), thickness vessel wall (TVW), and intima-media ratio (IMR) for the recipient (R) and transplant site (T) in arteries (A) and veins (V) were evaluated using H.E. staining. Flap loss (3%) was associated with increased ARED (<italic>p</italic> = 0.004) and ARID (<italic>p</italic> = 0.004). Preoperative radiotherapy led to a significant reduction in the outer diameter of the recipient vein in the neck (<italic>p</italic> = 0.018). Alcohol consumption (<italic>p</italic> = 0.05), previous thrombosis (<italic>p</italic> = 0.007), and diabetes (<italic>p</italic> = 0.002) were associated with an increase in the total thickness of venous recipient veins in the neck. Diabetes was also found to be associated with dilation of the venous media in the neck vessels (<italic>p</italic> = 0.007). The presence of cardiovascular disease (CVD) was associated with reduced intimal thickness (<italic>p</italic> = 0.016) and increased total venous vessel wall thickness (<italic>p</italic> = 0.017) at the transplant site. Revision surgeries were linked to increased internal and external diameters of the graft artery (<italic>p</italic> = 0.04 and <italic>p</italic> = 0.003, respectively), while patients with flap loss showed significantly increased artery diameters (<italic>p</italic> = 0.004). At the transplant site, alcohol influenced the enlargement of arm artery diameters (<italic>p</italic> = 0.03) and the intima-media ratio in the radial forearm flap (<italic>p</italic> = 0.013). In the anterolateral thigh, CVD significantly increased the intimal thickness and the intima-media ratio of the graft artery (<italic>p</italic> = 0.01 and <italic>p</italic> = 0.02, respectively). Patients with myocardial infarction displayed increased thickness in the <italic>A. thyroidea</italic> and artery media (<italic>p</italic> = 0.003). Facial arteries exhibited larger total vessel diameters in patients with CVD (<italic>p</italic> = 0.03), while facial arteries in patients with previous thrombosis had larger diameters and thicker media (<italic>p</italic> = 0.01). The presence of diabetes was associated with a reduced intima-media ratio (<italic>p</italic> < 0.001). Although the presence of diabetes, irradiation, and cardiovascular disease causes changes in vessel thickness in connecting vessels, these alterations did not adversely affect the overall success of the flap.


In vitro proliferation and osteogenic differentiation of endometrial stem cells and dental pulp stem cells.
Stem-cell-based therapies were introduced aiming to overcome the limitations of the existing procedures for regeneration of mineralized tissues. Stem cells isolated from the endometrial tissue and dental pulp have the capacity to differentiate into various functional cells including osteoblasts. However, studies comparing their ability to regenerate mineralized tissue are lacking. The purpose of this study was to compare the proliferation and osteogenic differentiation potential of endometrial stem cells (EnSCs) and dental pulp stem cells (DPSCs) using in vitro cell culture technique. The DPSCs and EnSCs were isolated from human dental pulp and endometrium, respectively. Their proliferation and osteogenic potential were compared in the same osteogenic medium (OM) after 3, 5, 7 and 10 days using the methyl thiazol tetrazolium assay, alizarin red staining, and real-time quantitative reverse transcription polymerase chain reaction (Real-Time qRT-PCR). The EnSCs showed higher proliferation rate compared to DPSCs. Regarding osteogenesis, alizarin red-positive colonies appeared earlier and in greater amounts in DPSCs group. The real-time qRT-PCR demonstrated significantly greater osteogenic potential of DPSCs compared to EnSCs. Our findings revealed significant differences in stem cell properties based on the tissue source. The EnSCs had greater proliferation rate than DPSCs, while DPSCs showed greater osteogenic potential compared to EnSCs in the same OM.


Effects of selected organic solvents on the astrocyte membrane ATPase in vitro.
1. The present study deals with astrocyte cultures as a model for studying the membrane-mediated central nervous system-depressing effect of organic solvents. 2. The primary astrocyte cultures were prepared from neonatal rat cerebella. The cells were cultured in modified essential medium. The astrocyte membranes isolated from the cultures were exposed to solvents in incubation mixture at different dose levels (3, 6 and 9 mmol/L) for 1 h. The physiologically important integral proteins Na+, K(+)-ATPase and Mg(2+)-ATPase were studied. 3. The aromatic hydrocarbons (benzene, toluene, styrene, xylene and ethylbenzene) inhibited the ATPase activities according to their lipid solubilities. n-Hexane and cyclohexane clearly had less effect than aromatic hydrocarbons, despite their greater lipid solubilities. 4. Astrocytes were shown to be sensitive targets to the effects of organic solvents, measured as the inhibition of the integral enzymes Na+, K(+)-ATPase and Mg(2+)-ATPase.


[Demonstration of an alkali PAS-effect using periodic acid at low concentration (author's transl)].
It has been shown that an isolated KOH PAS-effect of epithelial mucosubstances in the colonic mucosae of the rat can be demonstrated also without the conventionally used preliminary oxidation/reduction step. The method is based on the use of strongly diluted periodic acid after previous treatment of tissue sections with ethanolic KOH. As the positive reacting material has been proven to be sensitive against treatment with neuraminidase the modified KOH PAS-reaction should be related to the presence of acylated sialic acid residues in mucosubstances of the colonic mucosae of the rat.


Phenotypic and genotypic discrimination of Francisella tularensis ssp. holarctica clades.
Francisella tularensis is the causative agent of tularemia, a zoonotic disease with a wide host range. F. tularensis ssp. holarctica (Fth) is of clinical relevance for European countries, including Germany. Whole genome sequencing methods, including canonical Single Nucleotide Polymorphism (canSNP) typing and whole genome SNP typing, have revealed that European Fth strains belong to a few monophyletic populations. The majority of German Fth isolates belong to two basal phylogenetic clades B.6 (biovar I) and B.12 (biovar II). Strains of B.6 and B.12 seem to differ in their pathogenicity, and it has been shown that strains of biovar II are resistant against erythromycin. In this study, we present data corroborating our previous data demonstrating that basal clade B.12 can be divided into clades B.71 and B.72. By applying phylogenetic whole genome analysis as well as proteome analysis, we could verify that strains of these two clades are distinct from one another. This was confirmed by measuring the intensity of backscatter light on bacteria grown in liquid media. Strains belonging to clades B.6, B.71 or B.72 showed clade-specific backscatter growth curves. Furthermore, we present the whole genome sequence of strain A-1341, as a reference genome of clade B.71, and whole proteomes comparison of Fth strains belonging to clades B.6, B.71 and B.72. Further research is necessary to investigate phenotypes and putative differences in pathogenicity of the investigated different clades of Fth to better understand the relationship between observed phenotypes, pathogenicity and distribution of Fth strains.


Inhibition by dopamine of 86Rb efflux and hormone secretion from bovine anterior pituitary cells perifused in the presence of acetylcholine or TRH.
The effects of dopamine (10(-5) M) on growth hormone and prolactin secretion and the fractional rate of 86Rb efflux were investigated using perifused bovine pituitary cells. Dopamine decreased the control efflux rate, prevented the rise caused by thyroliberin (10(-7) M) in the presence of 3-isobutyl-1-methylxanthine (10(-4) M) and greatly decreased the rise caused by acetylcholine (2.5 x 10(-5) M). Under these conditions it inhibited the prolactin secretion but it had no effect on growth hormone secretion induced by either treatment. Dopamine receptors in lactotrophs are therefore coupled to secretion and potassium permeability, but if somatotrophs contain dopamine receptors they are coupled only to K+ efflux.


Exploring the relationship between eating disorder symptoms and substance use severity in women with comorbid PTSD and substance use disorders.
Eating disorders (ED) and substance use disorders (SUD) commonly co-occur, especially in conjunction with posttraumatic stress disorder (PTSD), yet little is known about ED and ED symptoms in women presenting to addiction treatment programs. We examined the association between ED symptoms and substance use frequency and severity in a sample of women with a DSM IV diagnosis of current SUD and PTSD enrolled in SUD treatment. Participants were 122 women from four substance abuse treatment sites who participated in a multi-site clinical trial through the National Institute of Drug Abuse Clinical Trials Network (NIDA CTN). The Eating Disorder Examination-Questionnaire (EDE-Q), the Clinician's Administered PTSD Scale (CAPS) and the Addiction Severity Index (ASI) were administered at baseline and correlational analyses were performed. Variables that significantly correlated with EDE-Q total and subscale scores were entered into a linear regression analysis. Scores on the EDE-Q Global scale, as well as the Eating Concern, Weight Concern and Shape Concern subscales of the EDE-Q were significantly associated with Caucasian race/ethnicity, past 30 day opiate use, higher ASI Psychiatric Subscale score and lower ASI Employment Subscale score. Although exploratory, these findings suggest that there may be a relationship between addiction severity, use of certain drugs of abuse and eating disorder symptoms, particularly those involving weight and shape concerns in women with comorbid PTSD and SUD.


Low-temperature thermal conductivity of superconductors with gap nodes.
We report a detailed analytic and numerical study of electronic thermal conductivity in d-wave superconductors. We compare theory of the crossover at low temperatures from T dependence to T(3) dependence for increasing temperature with recent experiments on YBa(2)Cu(3)O(7) in zero magnetic field for T approximately [0.04 K,0.4 K] by Hill et al. [Phys. Rev. Lett. 92, 027001 (2004)]. Transport theory, including impurity scattering and inelastic scattering within strong-coupling superconductivity, can consistently fit the temperature dependence of the data in the lower half of the temperature regime. We discuss the conditions under which we expect power-law dependences over wide temperature intervals.


Effect of operational strategies on activated sludge's acclimation to phenol, subsequent aerobic granulation, and accumulation of polyhydoxyalkanoates.
Aerobic granules, a relative novel form of microbial aggregate, are capable of degrading many toxic organic pollutants. Appropriate strategy is needed to acclimate seed sludge to the toxic compounds for successful granulation. In this study, two distinct strategies, i.e. mixed or single carbon sources, were experimented to obtain phenol-acclimated sludge. Their effects on reactor performance, biomass characteristics, microbial population and the granulation process were analyzed. Sludge fed with phenol alone exhibited faster acclimation and earlier appearance of granules, but possibly lower microbial diversity and reactor stability. Using a mixture of acetate and phenol in the acclimation stage, on the other hand, led to a reactor with slower phenol degradation and granulation, but eventual formation of strong and stable aerobic granules. In addition, the content of intracellular polyhydoxyakanoates (PHA) was also monitored, and significant accumulation was observed during the pre-granulation stage, where PHA >50% of dry weight was observed in both reactors.


Greater inhibitory effects of bivalirudin compared with unfractionated heparin plus eptifibitide on thrombin-induced platelet activation.
The effects of antithrombotic agents on the activation of platelets by thrombin were determined in blood from patients (n=12) with symptomatic coronary artery disease. Blood obtained immediately before cardiac catheterization was incubated in vitro with therapeutic concentrations of unfractionated heparin (1.2 and 2.0 U/ml) alone and in combination with eptifibatide (unfractionated heparin 1.2 U/ml+eptifibatide, 1.7 microg/ml) or bivalirudin (8 and 14 microg/ml). An activated clotting time was determined. Platelet activation was induced with thrombin (0, 5, 15, and 40 U/ml) and assessed with the use of flow cytometry. Fibrin polymerization was prevented by the peptide Gly-Pro-Arg-Pro. The activation of glycoprotein IIb-IIIa and surface expression of P-selectin were identified with antibodies (PAC-1 and anti-CD62). The activated clotting time was 258+/-13 s with 1.2 U/ml unfractionated heparin alone, 396+/-8 s with unfractionated heparin+eptifibatide, and 348+/-9 s with 8 microg/ml bivalirudin. The binding of PAC-1 (reflecting the potential to aggregate) was decreased most effectively by bivalirudin (percentage of platelets binding PAC-1 in response to 15 U/ml thrombin-unfractionated heparin=54+/-7%, unfractionated heparin+eptifibatide=12+/-4%, bivalirudin=1+/-0.3% of platelets, P<0.05 for bivalirudin compared with unfractionated heparin or unfractionated heparin+eptifibatide and unfractionated heparin+eptifibatide compared with unfractionated heparin). Bivalirudin prevented thrombin-induced surface expression of P-selectin more effectively than unfractionated heparin alone or unfractionated heparin+eptifibatide (percentage of platelets expressing P-selectin in response to 15 U/ml thrombin-unfractionated heparin alone=74+/-5%, unfractionated heparin+eptifibatide=53+/-7%, bivalirudin=1+/-0.3%, P<0.05 for bivalirudin compared with unfractionated heparin or unfractionated heparin+eptifibatide). Comparison between pharmacologic concentrations shown to be therapeutic demonstrated that bivalirudin inhibited thrombin-induced activation of platelets to a greater extent than unfractionated heparin alone or in combination with eptifibatide.


Glycosaminoglycan and glycoprotein synthesis by cranial neural crest cells in vitro.
In the developing chick embryo, cranial neural crest cells, that will subsequently give rise to facial mesenchyme tissues, migrate beneath the surface ectoderm in a cell-free and hyaluronate-rich matrix. To determine how the crest cells could contribute to this matrix, we cultured crest cells from stage 9 embryos for 2 days and then labeled them for 18--20 h with various precursors of glycosaminoglycans (GAG) and glycoproteins. [3H]fucose and [3H]glucosamine were incorporated into Pronase-sensitive macromolecules associated with the cell layer, but little labeled glycoprotein was released into the medium. Hyaluronate was the major GAG synthesized and was distributed between the cells and medium. Less chondroitin-sulfate was synthesized. In comparison, older cultures as well as fibroblasts produced different proportions of GAG. Our results confirm the autoradiographic findings of Pratt et al. (1975) and suggest that crest cells may contribute GAG to the matrix during migration.


Antigen-specific responses assessment for the evaluation of Bordetella pertussis T cell immunity in humans.
Measurement of antigen-specific T cell responses is an adjunctive parameter to evaluate protection induced by a previous Bordetella pertussis infection or vaccination. The assessment of T cell responses is technically complex and usually performed on fresh peripheral blood mononuclear cells (PBMC). The objective of this study was to identify simplified methods to assess pertussis specific T cell responses and verify if these assays could be performed using frozen/thawed (frozen) PBMC. Three read-outs to measure proliferation were compared: the fluorescent dye 5,6-carboxylfluorescein diacetate succinimidyl ester (CFSE) dilution test, the number of blast cells defined by physical parameters, and the incorporation of (3)H-thymidine. The results of pertussis-specific assays performed on fresh PBMC were compared to the results on frozen PBMC from the same donor. High concordance was obtained when the results of CFSE and blast read-outs were compared, an encouraging result since blast analysis allows the identification of proliferating cells and does not require any use of radioactive tracer as well as any staining. The results obtained using fresh and frozen PBMC from the same donor in the different T cell assays, including IFNγ and TNFα cytokine production, did not show significant differences, suggesting that a careful cryopreservation process of PBMC would not significantly influence T cell response evaluation. Adopting blast analysis and frozen PBMC, the possibility to test T cell responses is simplified and might be applied in population studies, providing for new instruments to better define correlates of protection still elusive in pertussis.


Innervation analysis of the small muscle bundles attached to the temporalis: truly new muscles or merely derivatives of the temporalis?
Detailed examinations were performed in ten temporal muscles from five cadavers to identify the muscle bundle arrangements of the temporalis and their innervation. Three additional muscle bundles were clearly observed in the main part of the fan-shaped temporalis: the anteromedial, anterolateral, and mid-lateral muscle bundles. Based on the origins, insertions and detailed innervation patterns, these bundles were considered as parts of the temporalis rather than independent muscles, although the anteromedial and anterolateral bundles had been recently reported as newly described muscles. A possible schematic model of the origins of these muscle bundles is proposed. We also report a branch from the posterior deep temporal nerve which was distributed to the temporal fascia and to the skin of the temporal region.


[Growth deficiency and renal tubular acidosis. The possible etiopathogenic mechanisms].
The authors have studied two cases of renal tubular acidosis in 2 children underlining that a possible defective mechanism of growth is based on alteration of the receptors of somatomedine C to level of cartilage. The deficit of receptors could be the consequence of the alterations of the acidosis on cartilage.


Effect of surround configuration on increment thresholds of a tiny violet flash.
Increment thresholds were measured for a 5', 435-nm test flash (50 or 200 msec) on background fields of various wavelengths and spatial configurations. As the field diameter was increased, the detection threshold for the 50-msec test decreased rapidly and leveled off when the field diameter exceeded 20'. For short-to-middle wavelength fields (much less than or equal to 503 nm), the detection threshold for the 200-msec test varied with field diameter in a similar manner to those observed for the 50-msec tests, while for middle-to-long wavelength fields (much greater than or equal to 544 nm), the detection threshold continued to decrease even when the field diameter exceeded 20'. The temporal-integration properties and the threshold vs intensity curves were determined for 465-nm and 579-nm fields to specify the mechanisms responsible for the detection. It was shown that the detection thresholds of the test upon the large 579-nm field were characterized by the short-wavelength mechanism isolated by an intense yellow adapting field, whereas the detection thresholds of the test upon the small 579-nm or the 465-nm field showed an intrusion of a more sensitive alternate mechanism, presumably the middle-wavelength mechanism. It was suggested that the sensitivity of the S cone signals might be affected by spatially distributed signals from the longer-wavelength mechanisms.


Central nervous system histoplasmosis with obstructive hydrocephalus.
A case of central nervous system histoplasmosis complicated by obstruction of the fourth ventricle is described. The patient rarely exhibited systemic symtoms of infection despite positive cultures for Histoplasma capsulatum from bone marrow, blood and urine. Infection recurred despite the administration of a total of 5 g of systemic amphotericin B. An additional course combined with intrathecal amphotericin B was terminated because of transverse myelitis.


Kainate receptor subunit 1 (GRIK1) risk variants and GRIK1 deficiency were detected in the Indian ADHD probands.
Executive dysfunctions caused by structural and functional abnormalities of the prefrontal cortex were reported in patients with Attention deficit hyperactivity disorder (ADHD). Owing to a higher expression of the glutamate ionotropic receptor kainate type subunit 1 (GluK1), encoded by the GRIK1 gene, in brain regions responsible for learning and memory, we hypothesized that GRIK1 might have a role in ADHD. GRIK1 variants rs363504 and rs363538, affecting the receptor function, were analyzed by case-control and family-based methods to identify the association with ADHD. The impact of these variants on ADHD-associated traits and pharmacological intervention were also analyzed. GRIK1 expression was quantified in the peripheral blood. The probands and their fathers had a higher frequency of rs363504 'CC' and rs363538 'CA' genotypes. Family-based investigation revealed maternal over transmission of rs363504 'C' and rs363538 'A' alleles to the probands. Quantitative trait analysis exhibited an association of rs363504 'TT' and rs363538 'AA' genotypes with higher hyperactivity scores of the probands. In the presence of rs363504 'TT' and rs363538 'CC' genotypes, MPH treatment improved hyperactivity and inattention, respectively. GRIK1 expression was significantly downregulated in the probands. We infer that GRIK1 affects ADHD etiology, warranting further in-depth investigation involving a larger cohort and more functional variants.


Development and validation of the Japanese version of the Restorative Sleep Questionnaire in community samples.
This study aimed to validate the Japanese version of the Restorative Sleep Questionnaire Weekly version (RSQ-W-J) and estimate the RSQ-W-J cut-off score to identify the degree of non-restorative sleep (NRS) that interferes with daytime function. An online survey was conducted, and 513 participants (241 men, 272 women; mean age, 47.88 ± 16.66 years) completed the questionnaires. The RSQ-W-J, Multidimensional Fatigue Inventory, Athens Insomnia Scale, Patient Health Questionnaire-9, Epworth Sleepiness Scale, Sleep Debt Index, NRS and the degree of daytime dysfunction, and the diagnostic criteria for chronic insomnia were used as implementation scales. The RSQ-W-J was confirmed to have a single-factor structure. Both the Cronbach α coefficient and test-retest reliability of RSQ-W-J were high (α = 0.91; intraclass correlation coefficient [ICC]agreement = 0.85). The RSQ-W-J was significantly negatively correlated with the scales of fatigue, insomnia, depressive symptoms, daytime sleepiness, and sleep debt. In addition, the RSQ-W-J scores of the insomnia group, in which participants met the diagnostic criteria for chronic insomnia, NRS group, in which participants had NRS only, and sleep debt group, in which participants had sleep debt, were significantly lower than those in the normal sleep group. Furthermore, the insomnia group scored significantly lower than the sleep debt group. The RSQ-W-J cut-off score, which indicates whether or not NRS interferes with daytime function, was estimated to be 45.8 points. The RSQ-W-J was shown to be useful for comprehensively measuring the degree of restorative sleep.


Development of genomic microsatellite markers for Aconitum gymnandrum (Ranunculaceae) by next generation sequencing (NGS).
Mating plays key roles in the demographic and genetic dynamics of populations. Estimates of mating portfolios and system based on progeny array (PA) method required highly polymorphic genetic markers, of which microsatellite is a good choice. In this study, we reported 19 polymorphic microsatellite loci for Aconitum gymnandrum. The number of alleles per locus ranged from 2 to 12. Observed and expected heterozygosity ranged from 0.000 to 1.000 and from 0.219 to 0.842, respectively. Seven loci showed significant deviation from Hardy-Weinberg equilibrium. These markers will provide a useful tool for pollination ecology and population genetic studies of A. gymnandrum in Qinghai-Tibet plateau.


Questioning the link between PTSD and cognitive dysfunction.
Posttraumatic stress disorder (PTSD) has been linked with impairment in cognitive functioning, but anomalies appear to arise on several levels, masking the true nature of the relationship. On one level, there is a blurring of the emotional and physical causes of cognitive dysfunction, especially with memory problems that are similar to those due to neurological disorders. At other levels, problems are evident due to issues such as overgeneralizing from specific populations to the general population, and the use of narrow-focused neuropsychological assessment instruments that neither dissociate among precise memory components nor relate to everyday situations. In this review we highlight methodological problems and concerns, and make suggestions to establish valid links between PTSD and cognitive dysfunction.


Analysis of receptor binding displacement curves by a nonhomologous ligand, on the basis of an equivalent competition principle.
An exact method for the analysis of receptor-ligand binding data when labeled bound ligand is displaced by a nonhomologous ligand with a different dissociation constant is described. The present method, which is based on an equivalent competition principle for the homologous and the nonhomologous ligand, converts displacement curves into a linear form and is also applicable to situations in which free concentrations of ligand are significantly smaller than the added concentrations as a result of ligand binding. It is shown that the dissociation constant of the nonhomologous ligand is given directly by the concentration of this nonhomologous ligand added and the free concentration of unlabeled homologous ligand required to give the same level of displacement of labeled bound ligand. On the basis of these displacement characteristics, all binding parameters for receptor interaction of the nonhomologous ligand can be obtained and expressed, for example, in a Scatchard plot. The present method, which is referred to as the equivalent competition method, is also evaluated in this study with respect to the effects of nonspecific ligand binding and the presence of multiple receptor classes.


Does muscle coactivation influence joint excursions during gait in children with and without hemiplegic cerebral palsy? Relationship between muscle coactivation and joint kinematics.
The theoretical role of muscle coactivation is to stiffen joints. The aim of this study was to assess the relationship between muscle coactivation and joint excursions during gait in children with and without hemiplegic cerebral palsy. Twelve children with hemiplegic cerebral palsy and twelve typically developing children underwent gait analysis at three different gait speeds. Sagittal hip, knee, and ankle kinematics were divided into their main components corresponding to joint excursions. A coactivation index was calculated for each excursion from the electromyographic envelopes of the rectus femoris/semitendinosus, vastus medialis/semitendinosus, or tibialis anterior/soleus muscles. Mixed linear analyses of covariance modeled joint excursions as a function of the coactivation index and limb. In typically developing children, increased coactivation was associated with reduced joint excursion for 8 of the 14 linear models (hip flexion, knee loading, knee extension in stance, knee flexion in swing, ankle plantarflexion from initial contact to foot-flat, ankle dorsiflexion in stance and in swing). Conversely, ankle plantarflexion excursion at push-off increased with increasing tibialis anterior/soleus coactivation. In the involved limbs of the children with cerebral palsy, knee loading, ankle plantarflexion at push off, and ankle dorsiflexion in swing decreased, while hip extension increased, with increasing muscle coactivation. The relationships between muscle coactivation and joint excursion were not equally distributed in both groups, and predominant in typically developing children. The results suggest that excessive muscle coactivation is not a cause of stiff-knee gait in children with hemiplegic cerebral palsy, but appears to be related to spastic drop foot.


Accuracy and sensitivity of high mobility group box 1 (HMGB1) in diagnosis of acute kidney injury caused by sepsis and relevance to prognosis.
The diagnostic value of high mobility group box 1 (HMGB1) levels in patients with acute kidney injury (AKI) caused by sepsis and its relationship with disease prognosis were investigated to improve patient survival. A total of 120 patients diagnosed with sepsis by comprehensive clinical examination were selected as the research subjects. According to the presence or absence of concurrent AKI, all patients were divided into SIAKI (50 cases with concurrent AKI) and N-AKI groups (70 cases without concurrent AKI). Sixty normal people receiving a physical examination in our hospital during the same period were divided into the control group. The diagnostic efficacy and the influences of HMGB1 on prognosis were assessed. HMGB1 levels in the serum and urine of the control group (3.43 ± 0.73 pg/mL, 343.13 ± 51.03 pg/mL) were both lower than those of the SIAKI (14.76 ± 2.44 pg/mL, 1109.76 ± 225.66 pg/mL) and N-AKI groups (7.99 ± 1.84 pg/mL, 890.54 ± 97.76 pg/mL) (P < 0.05). HMGB1 in the serum of the SIAKI group was higher than that of the N-AKI group (P < 0.05). The sensitivity (88%), specificity (87%), accuracy (88%), and area under the curve (AUC) (0.891) of the joint diagnosis of HMGB1 in blood and urine were superior to the diagnostic effects of HMGB1 in serum (70%, 70%, 70%, and 0.701) and HMGB1 in urine (59%, 57%, 58%, and 0.677) (P < 0.05). The proportion of HMGB1 in the nonsurvivors was higher than that in the survivors (85%) and was obviously higher than that in the survivors (15%) (P < 0.05). As a diagnostic marker of sepsis complicated with AKI, HMGB1 in serum and urine showed good application value. Serum HMGB1 could be used to assess disease prognosis with good clinical promotion.


The persistence of interleukin-6 is regulated by a blood buffer system derived from dendritic cells.
The interleukin-6 (IL-6) membrane receptor and its circulating soluble form, sIL-6R, can be targeted by antibody therapy to reduce deleterious immune signaling caused by chronic overexpression of the pro-inflammatory cytokine IL-6. This strategy may also hold promise for treating acute hyperinflammation, such as observed in coronavirus disease 2019 (COVID-19), highlighting a need to define regulators of IL-6 homeostasis. We found that conventional dendritic cells (cDCs), defined in mice via expression of the transcription factor Zbtb46, were a major source of circulating sIL-6R and, thus, systemically regulated IL-6 signaling. This was uncovered through identification of a cDC-dependent but T cell-independent modality that naturally adjuvants plasma cell differentiation and antibody responses to protein antigens. This pathway was then revealed as part of a broader biological buffer system in which cDC-derived sIL-6R set the in-solution persistence of IL-6. This control axis may further inform the development of therapeutic agents to modulate pro-inflammatory immune reactions.


Advances in understanding the association between Down syndrome and Hirschsprung disease (DS-HSCR).
The clinical association between Trisomy 21 (Down syndrome) and aganglionosis (Hirschsprung disease; DS-HSCR) is well-established, being of the order of 5% and remains the most common congenital association with Hirschsprung disease. However, little consensus exists as to the possible etiologic and genetic factors influencing this association. Recent research has identified a number of levels at which development of the enteric nervous system is potentially affected in Trisomy 21. These include a decreased central pool of available neuroblasts for migration into the enteric nervous system, abnormal neuroblast type, poor synaptic nerve function and early germline gene-related influences on the migrating neuroblasts due to genetic mutations of a number of important developmental genes, and possible somatic mutations resulting from alterations in the local tissue microenvironment. In this paper, we review available evidence for this association. In addition, we provide evidence of both germline and somatic gene mutations suggesting causation. Although the picture is complex, recent associations between specific RET proto-oncogene variations have been shown to be significant in Down syndrome patients with Hirschsprung disease, as they probably interfere with vital RET functions in the development of the autonomic and enteric nervous systems, increasing the risk of disturbed normal function. In addition, we explore potential role of other facilitatory influence of other susceptibility genes as well as potential other chromosome 21 gene actions and the microenvironment on the Down syndrome gastro-intestinal tract. The various ways in which trisomy of chromosome influences the enteric nervous system are becoming clearer. The sum of these effects influences the outcome of surgery in Down syndrome patients with Hirschsprung Disease.


Characterization of mass transfer of lower chlorinated benzenes from contaminated sediment into water.
Mass transport of chlorinated benzenes as found at the Petro-Processors of Louisiana, Inc. (PPI) Superfund site was characterized for a range of flow rates in small streams. At this site hazardous waste was historically disposed of in unengineered pits. Hexachlorobenzene and lesser chlorinated degradation products were found among other compounds. As waste was being disposed into unengineered pits, it seeped to lagoons and sediments of Baton Rouge Bayou (BRB), which flows through and nearby the former disposal areas. Characterization of the transport and fate of chlorobenzenes at PPI is an integral part of the Monitored Natural Attenuation (MNA) remedy currently underway. Laboratory experimental results and mathematical model predictions of the flux of 1,3-dichlorobenzene (1,3-DCB) from sediments into water are presented. 1,3-DCB was studied as individually and as part of a mixture of four contaminants with 1,2-DCB, chlorobenzene (MCB) and trichlorobenzene (TCB). Surficial sediments were collected, spiked with contaminants and leached to determine flux over time. Two advection-dispersion models were tested and the effect of low, cycling and fast stream flow on the contaminant flux was assessed. Model results suggest that tortuosity and effective diffusivity are related effective system predictors and descriptors. Statistical analysis supports the models' predictive capabilities.


Association between cholinesterase inhibitors and kidney function decline in patients with Alzheimer's dementia.
Preclinical evidence shows that activation of the cholinergic anti-inflammatory pathway (CAP) may have direct and indirect beneficial effects on the kidney. Cholinesterase inhibitors (ChEIs) are specific Alzheimer's dementia (AD) therapies that block the action of cholinesterases and activate CAP. Here, we explored a plausible effect of ChEIs on slowing kidney function decline by comparing the risk of CKD progression among patients with newly diagnosed AD that initiated ChEI or not within 90 days. Using complete information of routine serum creatinine tests, we evaluated changes in estimated glomerular filtration rate (eGFR) and defined the outcome of chronic kidney disease (CKD) progression as the composite of an eGFR decline of over 30%, initiation of dialysis/transplant or death attributed to CKD. A secondary outcome was death. Inverse probability of treatment-weighted Cox regression was used to estimate hazard ratios. Among 11, 898 patients, 6,803 started on ChEIs and 5,095 did not. Mean age was 80 years (64% women) and the mean eGFR was 68 ml/min/1.73m2. During a median 3.0 years of follow-up, and compared to non-use, ChEI use was associated with 18% lower risk of CKD progression (1,231 events, adjusted hazard ratio 0.82; 95% confidence interval 0.71-0.96) and a 21% lower risk of death (0.79; 0.72-0.86). Results were consistent across subgroups, ChEI subclasses and after accounting for competing risks. Thus, in patients with AD undergoing routine care, use of ChEI (vs no-use) was associated with lower risk of CKD progression.


Correlations between periodontitis and loss of mandibular bone in relation to systemic bone changes in postmenopausal Japanese women.
A new method of measuring mandibular alveolar bone mineral density (BMD) was applied to 40 postmenopausal Japanese women aged 50-69 years exhibiting minimal to mild periodontal diseases. Lumbar spine BMD was measured by dual X-ray absorptiometry (DXA) and calcaneus speed of sound (SOS) by quantitative ultrasound (QUS). There were age-related decreases of alveolar BMD, calcaneus SOS and vertebral BMD. There were significant correlations between two of the respective bone mass values. Correlations between clinical dental findings and bone mass data including alveolar BMD, SOS and lumbar spine BMD were investigated. Significant correlations were demonstrated between alveolar BMD and calcaneus SOS or vertebral BMD. Alveolar BMD showed significant correlation with clinical dental findings including periodontal pocket depth and mobility as well as calcaneus SOS and lumbar spine BMD. Using multivariate analysis combinations of univariate predictors, including deoxypyridinoline (DPD), significantly predicted attachment levels. The SOS value was useful combined with other predictors for predicting attachment level. It was concluded that the new method of evaluating alveolar BMD is useful to predict systemic bone mass and strength as well as dental clinical findings.


3-Methylglutaric acid in energy metabolism.
3-methylglutaric (3MG) acid is a conspicuous C6 dicarboxylic organic acid classically associated with two distinct leucine pathway enzyme deficiencies. 3MG acid is excreted in urine of individuals harboring deficiencies in 3-hydroxy-3-methylglutaryl (HMG) CoA lyase (HMGCL) or 3-methylglutaconyl CoA hydratase (AUH). Whereas 3MG CoA is not part of the leucine catabolic pathway, it is likely formed via a side reaction involving reduction of the α-ß trans double bond in the leucine pathway intermediate, 3-methylglutaconyl CoA. While the metabolic basis for the accumulation of 3MG acid in subjects with deficiencies in HMGCL or AUH is apparent, the occurrence of 3MG aciduria in a host of unrelated inborn errors of metabolism associated with compromised mitochondrial energy metabolism is less clear. Herein, a novel mitochondrial biosynthetic pathway termed "the acetyl CoA diversion pathway", provides an explanation. The pathway is initiated by defective electron transport chain function which, ultimately, inhibits acetyl CoA entry into the TCA cycle. When this occurs, 3MG acid is synthesized in five steps from acetyl CoA via a novel reaction sequence, providing a metabolic rationale for the connection between 3MG aciduria and compromised mitochondrial energy metabolism.


Effect of aminophylline upon the protective activity of common antiepileptic drugs and their plasma levels in mice.
Aminophylline (50 mg/kg) decreased the protective efficacy of carbamazepine (20 mg/kg), diphenylhydantoin (8-12 mg/kg), phenobarbital (20 and 25 mg/kg), and valproate (250 and 300 mg/kg) against electroconvulsions in mice. On the other hand, aminophylline (5 mg/kg) was devoid of such activity. Plasma levels of antiepileptic drugs were measured with the help of the Abbott TDx analyzer and after administration of carbamazepine (20 mg/kg), diphenylhydantoin (10 mg/kg), phenobarbital (25 mg/kg), and valproate (250 mg/kg) were as follows: 8.61, 6.48, 24.3 and 329 micrograms/ml, respectively. Aminophylline (50 mg/kg) remained without any significant influence upon these plasma levels. This may lead to the conclusion that aminophylline-induced reversal of antiepileptic drug activity is not dependent upon a pharmacokinetic mechanism and probably occurs at the neuronal level.


Single vs multivessel treatment during primary angioplasty: results of the multicentre randomised HEpacoat for cuLPrit or multivessel stenting for Acute Myocardial Infarction (HELP AMI) Study.
Prospective randomized, multicentre study. Recanalisation of the culprit lesion is the main goal of primary angioplasty for acute myocardial infarction. With the exception of cardiogenic shock, staged procedures are performed in the presence of multivessel disease. The study hypothesis is that with modern non-thrombogenic stents (heparin coated) complete revascularization with multivessel treatment can be safely achieved during the primary angioplasty procedure with a lower need of subsequent revascularization procedures and at a lower cost. PRIMARY: 12-month incidence of repeat revascularization (any revascularization, infarct related artery as well as non-infarct-related artery). SECONDARY: (1) in hospital repeat revascularization, reinfarction and death; (2) total hospital cost (including a 12 months follow-up period). 69 patients with ST elevation Acute Myocardial Infarction (AMI), <12 hours after symptoms onset, undergoing primary angioplasty, with documented multivessel disease and both culprit lesion and 1 to 3 other lesions suitable for stent implantation. Unbalanced randomization between culprit lesion treatment only (n = 17) and complete multivessel treatment (n = 52, with 71 additional lesions treated). The two groups were well balanced in terms of clinical characteristics, number of diseased vessels and angiographic characteristics of the culprit lesion. In the complete multivessel treatment group 2.36 +/- 0.64 lesions per patient were treated using 2.73 +/- 0.78 heparin coated stents (1.00 lesions and 1.29 +/- 0.61 stents in the culprit treatment group, both p < 0.001). The duration of the procedure increased from 53 +/- 21 min (culprit treatment group) to 69 +/- 32 min (p = 0.032) and the amount of contrast used from 242 +/- 102 ml (culprit treatment group) to 341 +/- 163 ml (multivessel complete treatment), p = 0.025. A similar low incidence of in-hospital major adverse cardiac events was observed in the 2 groups (0 and 3.8% in culprit and multivessel treatment groups, p = 0.164). The increase in the incidence of new revascularisation in the culprit treatment group at 12 month follow-up was not significant (35 vs 17%, p = 0.247) but was sufficient to compensate the initial higher in-hospital cost, with a similar 12 month hospital cost in the 2 groups (Euro 22,330 +/- Euro 13,653 vs Euro 20,382 +/- Euro 11,671, p = 0.231). Multivessel treatment during primary PTCA was safe in this controlled trial. However, when only the culprit lesion was initially treated, the need for subsequent clinically driven revascularization remained low and no clinical or economical advantages were obtainable with a more aggressive initial approach. In clinical practice, a staged approach to multivessel treatment during primary angioplasty avoids to treat unnecessarily non clinically relevant lesions.


An evidence-based study on the current status of Chinese secondary school mathematics teachers' autonomous learning capacity across demographic and contextual factors.
Autonomous learning capacity is a key competency that supports teachers' professional development. In this study, a stratified sampling method was used to recruit 396 junior and senior high school mathematics teachers in T city, one of the provincial city in China. A questionnaire with high reliability and validity developed prior to the study by the researchers was employed to measure their autonomous learning capacity and differences across groups. Twelve teachers were then selected for interviews. The results showed satisfactory overall performance. By subdimension, teachers' performance was the best in the development of study plans, followed by evaluation of learning outcomes, while they needed improvement in learning habit formation and proficiency in using learning methods. Furthermore, the analysis of differences across groups indicated that for autonomous learning capacity, female teachers were significantly better than their male peers; junior high school teachers were better compared to those in senior high school; teachers aged 41-50 underperformed those aged 51 and above; teachers who work in rural areas and townships did not perform worse than urban teachers; and those with doctoral degrees did not demonstrate considerable advantage over others. There were no significant differences in the overall autonomous learning capacity across years of teaching and job title groups. However, in the subdimensions, those with 21-30 years of teaching experience had lower proficiency in using learning methods and evaluation of learning outcomes, and teachers with senior titles did not demonstrate expected advantages in learning habit formation.


Arginine Biosynthesis by a Bacterial Symbiont Enables Nitric Oxide Production and Facilitates Larval Settlement in the Marine-Sponge Host.
Larval settlement and metamorphosis are regulated by nitric oxide (NO) signaling in a wide diversity of marine invertebrates.1-10 It is thus surprising that, in most invertebrates, the substrate for NO synthesis-arginine-cannot be biosynthesized but instead must be exogenously sourced.11 In the sponge Amphimedon queenslandica, vertically inherited proteobacterial symbionts in the larva are able to biosynthesize arginine.12,13 Here, we test the hypothesis that symbionts provide arginine to the sponge host so that nitric oxide synthase expressed in the larva can produce NO, which regulates metamorphosis,8 and the byproduct citrulline (Figure 1). First, we find support for an arginine-citrulline biosynthetic loop in this sponge larval holobiont by using stable isotope tracing. In symbionts, incorporated 13C-citrulline decreases as 13C-arginine increases, consistent with the use of exogenous citrulline for arginine synthesis. In contrast, 13C-citrulline accumulates in larvae as 13C-arginine decreases, demonstrating the uptake of exogenous arginine and its conversion to NO and citrulline. Second, we show that, although Amphimedon larvae can derive arginine directly from seawater, normal settlement and metamorphosis can occur in artificial sea water lacking arginine. Together, these results support holobiont complementation of the arginine-citrulline loop and NO biosynthesis in Amphimedon larvae, suggesting a critical role for bacterial symbionts in the development of this marine sponge. Given that NO regulates settlement and metamorphosis in diverse animal phyla1-10 and arginine is procured externally in most animals,11 we propose that symbionts might play an equally critical regulatory role in this essential life cycle transition in other metazoans.


Axonal injury following mild traumatic brain injury is exacerbated by repetitive insult and is linked to the delayed attenuation of NeuN expression without concomitant neuronal death in the mouse.
Mild traumatic brain injury (mTBI) affects brain structure and function and can lead to persistent abnormalities. Repetitive mTBI exacerbates the acute phase response to injury. Nonetheless, its long-term implications remain poorly understood, particularly in the context of traumatic axonal injury (TAI), a player in TBI morbidity via axonal disconnection, synaptic loss and retrograde neuronal perturbation. In contrast to the examination of these processes in the acute phase of injury, the chronic-phase burden of TAI and/or its implications for retrograde neuronal perturbation or death have received little consideration. To critically assess this issue, murine neocortical tissue was investigated at acute (24-h postinjury, 24hpi) and chronic time points (28-days postinjury, 28dpi) after singular or repetitive mTBI induced by central fluid percussion injury (cFPI). Neurons were immunofluorescently labeled for NeuroTrace and NeuN (all neurons), p-c-Jun (axotomized neurons) and DRAQ5 (cell nuclei), imaged in 3D and quantified in automated manner. Single mTBI produced axotomy in 10% of neurons at 24hpi and the percentage increased after repetitive injury. The fraction of p-c-Jun+ neurons decreased at 28dpi but without neuronal loss (NeuroTrace), suggesting their reorganization and/or repair following TAI. In contrast, NeuN+ neurons decreased with repetitive injury at 24hpi while the corresponding fraction of NeuroTrace+ neurons decreased over 28dpi. Attenuated NeuN expression was linked exclusively to non-axotomized neurons at 24hpi which extended to the axotomized at 28dpi, revealing a delayed response of the axotomized neurons. Collectively, we demonstrate an increased burden of TAI after repetitive mTBI, which is most striking in the acute phase response to the injury. Our finding of widespread axotomy in large fields of intact neurons contradicts the notion that repetitive mTBI elicits progressive neuronal death, rather, emphasizing the importance of axotomy-mediated change.


Cholinergic excitation of dopaminergic cells depends on sequential activation of protein kinase C and the L-type calcium channel in ventral tegmental area slices.
Dopaminergic projections from the ventral tegmental area (VTA) constitute the mesolimbocortical system that underlies addiction and psychosis primarily as the result of increased dopaminergic transmission. Dopaminergic neurons in the VTA receive glutamatergic and cholinergic innervations that regulate their firing activities. Both transmitter systems can activate protein kinase C (PKC) by increasing intracellular calcium and lipid second messengers, however, whether PKC mediates increased firing following glutamatergic and cholinergic activation remains unknown. This paper examined the effects of acute PKC inhibition on firing responses to carbachol, NMDA or AMPA using patch clamp recordings from brain slices. The three ligands all induced a reversible increase in firing, however, only carbachol-induced increase in firing was attenuated by the PKC inhibitors chelerythrine or GF 109203X. The L-type calcium channel blocker nifedipine partially blocked carbachol-induced excitation similar to PKC inhibitors. PKC inhibition and L-type channel blockade did not significantly alter NMDA- or AMPA-induced excitation. Concurrent blockade of PKC and L-type channels with chelerythrine and nifedipine did not additively suppress carbachol-induced excitation indicating they were sequential events in the same signaling pathway. Furthermore, preincubation with the PKC inhibitor GF 109203X reduced the carbachol-induced increase in nifedipine-sensitive high-voltage gated calcium currents. These results indicate that cholinergic activation enhances PKC activity, which in turn facilitates L-type channel opening to excite dopaminergic cells, a finding that is in line with reports of increased PKC in the VTA in animals displaying addictive behavior.


A comparison of directed functional connectivity among fist-related brain activities during movement imagery, movement execution, and movement observation.
Brain-computer interface (BCI) has been widely used in sports training and rehabilitation training. It is primarily based on action simulation, including movement imagery (MI) and movement observation (MO). However, the development of BCI technology is limited due to the challenge of getting an in-depth understanding of brain networks involved in MI, MO, and movement execution (ME). To better understand the brain activity changes and the communications across various brain regions under MO, ME, and MI, this study conducted the fist experiment under MO, ME, and MI. We recorded 64-channel electroencephalography (EEG) from 39 healthy subjects (25 males, 14 females, all right-handed) during fist tasks, obtained intensities and locations of sources using EEG source imaging (ESI), computed source activation modes, and finally investigated the brain networks using spectral Granger causality (GC). The brain regions involved in the three motor conditions are similar, but the degree of participation of each brain region and the network connections among the brain regions are different. MO, ME, and MI did not recruit shared brain connectivity networks. In addition, both source activation modes and brain network connectivity had lateralization advantages.


The impact of COVID-19 on cervicofacial infection of dental aetiology.
Cervicofacial infections of dental aetiology can be life-threatening and with the closure of dental practices following the onset of the COVID-19, it would be anticipated that their prevalence presenting to maxillofacial surgery would increase and services may be overwhelmed, with patients presenting later with a potential subsequent increase in morbidity. A retrospective analysis of patients with cervicofacial infection of dental aetiology referred to maxillofacial surgery during the initial six weeks of COVID-19 lockdown in 2020 was carried out and compared with the equivalent period in the two preceding years. Unexpectedly, during COVID-19 lockdown, there was a reduction in patients seen with cervicofacial infection of dental aetiology. This may have resulted from patient adherence to government guidelines "Stay at home", successful triaging of patients in primary care and emergency treatment provided by urgent dental care centres. Proportionally more patients who presented to hospital had received prior antibiotic therapy and required in-patient admission. All patients admitted received incision and drainage, with an increase extraoral drainage and an associated reduction in length of stay. During COVID-19 lockdown, maxillofacial managed a reduced number of patients with cervicofacial infection, likely resulting from primary and secondary dental care working together. The rate of incision and drainage of patients not admitted increased under local anaesthesia with increase of extraoral drainage and reduced length of stay for those admitted.


Hypothalamic energy metabolism is impaired by doxorubicin independently of inflammation in non-tumour-bearing rats.
We sought to explore the effects of doxorubicin on inflammatory profiles and energy metabolism in the hypothalamus of rats. To investigate these effects, we formed two groups: a control (C) group and a Doxorubicin (DOXO) group. Sixteen rats were randomly assigned to either the control (C) or DOXO groups. The hypothalamus was collected. The levels of interleukin (IL)-1β, IL-6, IL-10, TNF-α and energy metabolism (malate dehydrogenase, complex I and III activities) were analysed in the hypothalamus. The DOXO group exhibited a decreased body weight (p < 0.01). Hypothalamic malate dehydrogenase activity was reduced when compared with control (p < 0.05). In addition, pro-inflammatory cytokine levels were unchanged. Therefore, our results demonstrate that doxorubicin leads to an impairment of \hypothalamic energy metabolism, but do not affect the inflammatory pathway. SIGNIFICANCE PARAGRAPH: The hypothalamus is a central organ that regulates a great number of functions, such as food intake, temperature and energy expenditure, among others. Doxorubicin can lead to deep anorexia and metabolic chaos; thus, we observed the effect of this chemotherapeutic drug on the inflammation and metabolism in rats after the administration of doxorubicin in order to understand the central effect in the hypothalamus. Drug treatment by doxorubicin is used as a cancer therapy; however the use of this drug may cause harmful alterations to the metabolism. Thus, further investigations are needed on the impact of drug therapy over the long term.


Civil commitment: a review of traditional arrangements in the provision of involuntary hospitalization of the mentally ill.
The history of civil commitment procedures is reviewed, with an analysis of the origins of commitment authority. Views contrary to involuntary hospitalization are presented. The author argues for the retention of these procedures, with an emphasis upon the parens patriae approach.


Neural correlates for aesthetic appraisal of pictograph and its referent: An fMRI study.
As one kind of language symbols, pictograph has a graphical structure based on its referential object. Are the aesthetic qualities of these referential objects reflected in the cognitive and neural processing of the font structure of pictographs? In this present study, participants performed aesthetic judgments on both pictographs and their referential object images whilst acquiring fMRI data. Square luminance judgment was served as baseline to control for activity in motor brain regions associated with the key responses. Behavioral results showed both font structures of pictographs and their corresponding object images which refer to beautiful objects were rated significantly higher on beauty than those refer to ugly objects. Functional MRI revealed regions of occipital lobe, frontal lobe and inferior orbitofrontal cortex (OFC) that were commonly activated for pictographs and object images referring to beautiful objects in the direct contrasts between beautiful judgment and luminance judgment. Moreover, only the aesthetic judgments of beauty for the pictographs but not the object images elicited activation of motor areas, which implied relevant embodied experience was elicited during the aesthetic perception of novel pictographs. In contrast, activities associated with pictographs referring to ugly objects were limited to visual processing regions of the bilateral inferior occipital gyri, which could be due to the intentions of avoiding ugly or aversive stimulus. In addition, object images were associated with broader recruitment in cortical areas than pictographs, likely due to the increased visual processing demands for the more visually complex object images. These findings indicate that aesthetic perceptions for the font structures of pictographs were derived from their referential objects, which could arouse a similar sense of beauty supported by common neural mechanisms with other aesthetic objects.


Detrusor hyperreflexia: the effect of posture and pelvic floor activity.
Detrusor hyperreflexia is described in patients with neurological disorders, bladder outflow obstruction, persistent enuresis and in female incontinent patients with obvious neurological or urological disease. Postural changes were present in 60% of the patients and in 45% of the patients examined reflex relaxation of the pelvic floor occurred with the uninhibited detrusor contractions.


Focal inflammation in the embryological ventral pancreas: assessment using CT and MRI.
To identify the characteristic computed tomography (CT) and magnetic resonance imaging (MRI) findings of nonuniform inflammation between the posterior aspect of the head and the remainder of the pancreas. Two radiologists retrospectively evaluated images of 19 patients in whom the degree of attenuation in the CT images and/or the signal intensity in the MRI images differed between the posterior aspect of the head (ventral primordium) and the remainder of the pancreas (dorsal primordium) due to benign disorders other than uneven fatty replacement. Multiphase, contrast-enhanced CT examinations were performed in 17 patients. In five, T1- and T2-weighted images with and without fat suppression were obtained using a 1.5 T superconducting MRI system. The lesions were localized in the posterior aspect of the head in 17 patients. In two patients, the lesions occupied the posterior aspect of the head and extended to the neck. The lesions exhibited the following findings: inhomogeneous hypoattenuation (with spotty areas of relatively well-maintained contrast enhancement) during the pancreatic phase (17/17) and no intense peripheral enhancement or central necrotic areas (17/17) in CT images, hypointensity in T1-weighed images (5/5), hyperintensity in T2-weighed images (4/5), no vascular invasion (19/19), failure to depict the entire course of Wirsung's duct (17/19), and calcification (13/19). Santorini's ducts were depicted in all but three patients. Focal inflammation in the embryological ventral pancreas exhibits a unique anatomical distribution and characteristic findings.


Use of a combination of chemotherapy and radiation therapy in the management of advanced basal cell carcinoma of the head and neck.
A series of patients with massive rodent-type basal cell carcinoma of the head are presented as having therapeutic responses with chemotherapy combined with radiation therapy ranging from partial to complete response. No cases were cured by the regimen. The nature of large basal cell carcinomas of the head and treatment philosophy for surgically unresectable tumor are discussed.


[The status of acquiring written informed consent to clinical practice of medical students in our department of anesthesiology].
There is no information on the acquisition status of the written informed consent to clinical anesthesia practice of medical students. The contents of the clinical practice were divided into three categories: No.1 (the minor procedures such as mask-and-bag ventilation, blood drawing, drug administration), No.2 (only the visit), No.3 (non-cooperation). We asked the patient to chose one of them. From the written informed consents given by 1531 patients scheduled for operation, we analyzed the degree of the cooperation and the relation between the cooperation and the patient background. Although 78% of patients agreed to cooperate in some way (No.1 or No.2), there were only 15% of patients who allowed students to perform minor procedures. The results showed that clinical practice in anesthesiology can not be performed properly by only the visit. For a meaningful clinical practice in anesthesiology to be performed, it will be useful to use high-fidelity human patient simulator (HF-PS) actively and campaign for the cooperation and the understanding of the general pablic about medical education.


Digital object attachment.
Digital object attachment is a complex phenomenon that has received little attention in the extant literature. The purpose of the current article is to develop a conceptual model to clarify discrepancies and encourage future work in this area. We classify digital objects into four categories by their complexity, user control, interactivity and user value. The proposed model captures the dynamic and cyclical interplay between the user, the digital object and the environment. The user-object relationship can evolve over time through user dedication and psychological ownership, enabling identity experimentation and formation that ultimately lead to object attachment. In advanced forms, digital objects can assume transitional properties that blur the user's offline and digital existence and trigger identity transference.


[Changes in the microcirculation in the rat mesentery during hypotension induced by adenosine].
In anesthetized rats, i.v. administration of adenosine (2.5 mg/kg/min) for 30 min caused reduction of mean systemic blood pressure. A significant decrease of blood flow was found in mesentery microvessels as well as dilation of arterioles and venules.


E3 ubiquitin ligase-mediated regulation of bone formation and tumorigenesis.
The ubiquitination-proteasome and degradation system is an essential process that regulates protein homeostasis. This system is involved in the regulation of cell proliferation, differentiation and survival, and dysregulations in this system lead to pathologies including cancers. The ubiquitination system is an enzymatic cascade that mediates the marking of target proteins by an ubiquitin label and thereby directs their degradation through the proteasome pathway. The ubiquitination of proteins occurs through a three-step process involving ubiquitin activation by the E1 enzyme, allowing for the transfer to a ubiquitin-conjugated enzyme E2 and to the targeted protein via ubiquitin-protein ligases (E3), the most abundant group of enzymes involved in ubiquitination. Significant advances have been made in our understanding of the role of E3 ubiquitin ligases in the control of bone turnover and tumorigenesis. These ligases are implicated in the regulation of bone cells through the degradation of receptor tyrosine kinases, signaling molecules and transcription factors. Initial studies showed that the E3 ubiquitin ligase c-Cbl, a multi-domain scaffold protein, regulates bone resorption by interacting with several molecules in osteoclasts. Further studies showed that c-Cbl controls the ubiquitination of signaling molecules in osteoblasts and in turn regulates osteoblast proliferation, differentiation and survival. Recent data indicate that c-Cbl expression is decreased in primary bone tumors, resulting in excessive receptor tyrosine kinase signaling. Consistently, c-Cbl ectopic expression reduces bone tumorigenesis by promoting tyrosine kinase receptor degradation. Here, we review the mechanisms of action of E3 ubiquitin ligases in the regulation of normal and pathologic bone formation, and we discuss how targeting the interactions of c-Cbl with some substrates may be a potential therapeutic strategy to promote osteogenesis and to reduce tumorigenesis.


Kinetics of gene expression of alkaline phosphatase during healing of alveolar bone in rats.
Immunohistochemical studies and molecular biology have enabled us to identify numerous proteins that are involved in the metabolism of bone, and their encoding genes. Among these is alkaline phosphatase (ALP), an enzyme that is responsible for the initiation of mineralisation of the extracellular matrix during alveolar bone repair. To evaluate the gene expression of ALP during this process, we studied nine healthy adult male rats, which had their maxillary central incisors extracted from the right side and were randomly divided into three groups. During three experimental periods, 7 days, 14 days, and 28 days, the alveoli were curetted, the rats killed, and samples analysed by real-time reverse transcription polymerase chain reaction (qRT-PCR). The RNAm that encodes the gene for the synthesis of ALP was expressed during the three periods analysed, but its concentration was significantly increased at 14 and 28 days compared with at 7 days. There was no significant difference between 14 and 28 days (p=0.0005). We conclude that genes related to ALP are expressed throughout the healing process and more intensively during the later periods (14 and 28 days), which coincides with the increased formation of mineralised bone.


CIITA-regulated plexin-A1 affects T-cell-dendritic cell interactions.
The major histocompatibility complex (MHC) class II transactivator (CIITA) is the 'master coactivator' of MHC class II genes. To identify new targets of CIITA, we analyzed cDNA microarrays of dendritic cells (DCs) from CIITA-deficient, MHC class II-deficient and control mice. We found the semaphorin receptor plexin-A1 was expressed in DCs, but not in other immune cells, and was strongly induced by CIITA. RNA interference by short hairpin RNA specific for plexin-A1, but not a single-nucleotide mutant, greatly reduced plexin-A1 expression and T cell stimulation by protein- or peptide-antigen-pulsed DCs.Plexin-A1 is not required for peptide binding to MHC. These data indicate involvement of plexin-A1 in T cell-DC interactions but not antigen processing or binding.


Metitepine distinguishes two receptors mediating inhibition of [3H]-5-hydroxytryptamine release in guinea pig hippocampus.
Inhibition of [3H]-5-hydroxytryptamine ([3H]-5-HT) release from guinea pig brain slices via activation of the terminal 5-HT autoreceptor has previously been characterised as a model of 5-HT1D receptor activation, based on the rank potencies of a range of agonists, and the potent antagonism of the inhibitory effects of 5-HT by metitepine. The present study uses this model, in slices of the guinea pig hippocampus, to examine the antagonist potency of metitepine against the 5-HT receptor agonists sumatriptan, 5-carboxamidotryptamine (5-CT) and 5-HT. Addition of metitepine to the perfusion buffer (30, 300 and 1000 nmol/l) significantly shifted the concentration-response curve to 5-HT, producing a Schild slope of 1.1, and a pA2 value of 7.6. However, the ability of metitepine to antagonise the effects of sumatriptan or 5-CT in this model was less marked. A clear-cut shift in the concentration-response curve to sumatriptan was only achieved at 1000 nmol/l metitepine (apparent pA2 = 6.7), and this was similar to the ability of metitepine to attenuate the effects of 5-CT (apparent pA2 7.0 at 300 nmol/l and 6.7 at 1000 nmol/l). These findings suggest heterogeneity in the receptor mediating inhibition of [3H]-5-HT release in guinea pig hippocampus.


Nonstandard imaging methods in electron microscopy.
A rigorous treatment of image formation, taking into account inelastic scattering and partially coherent illumination, is given for the CTEM. The results also hold for a STEM if the momentum vectors of the incident and outgoing waves are interchanged and the flight direction of the electrons is reversed. A generalized optical theorem is found which connects the anti-Friedel part of the elastic scattering amplitude with the squares of the elastic and inelastic scattering amplitudes. This theorem has important consequences for all approximation methods. For example, it shows that inelastic scattering influences the elastic scattering amplitude. It is further shown that the observed wings in optical diffractograms, obtained from micrographs of amorphous material taken using tilted beam illumination, can only be explained if double scattering (elastic and inelastic) is assumed. Hollow-cone illumination strongly suppresses the structural noise in the phase contrast image. The corresponding phase contrast transfer function (PCTF) is calculated for two different cone angles, assuming that no beam-limiting aperture is present. In addition, the PCTFs of a corrected microscope are calculated for two different illumination modes, one yielding negative and the other positive phase contrast. Finally, the contrast transfer has been investigated for a stem operating with a split detector.


Cross-bridge behaviour in skinned smooth muscle of the guinea-pig taenia coli at altered ionic strength.
1. The effects of varied levels (25-300 mM) of ionic strength on mechanical properties and ATP hydrolysis rate of chemically skinned guinea-pig taenia coli fibres were investigated. 2. The tension development following activation by calcium (pCa 4.8), and relaxation following removal of calcium (pCa 9), were slower in 25 mM compared to 150 mM ionic strength. In fibres activated by thiophosphorylation of myosin light chains, by exposure to ATP-gamma-S, the tension development was rapid and independent of ionic strength. 3. The maximal shortening velocity (Vmax) was obtained from force-velocity relations determined by the quick-release method. The rate of ATP hydrolysis (JATP) was determined by measurement of pyruvate released from phosphoenolpyruvate (PEP). In order to obtain maximal Vmax and JATP at a Mg-ATP concentration of 1 mM, an ATP regenerating system was required. In thiophosphorylated fibres 2 mM-phosphocreatine (PCr) or 3.2 mM-PEP were adequate for maximal Vmax and JATP respectively. In calcium-activated fibres 5 mM-PCr was required for maximal Vmax. 4. The isometric force of thiophosphorylated fibres showed a biphasic dependence on ionic strength with a maximum at 90 mM. Vmax was essentially unchanged between 50 and 200 mM ionic strength. At 25 mM ionic strength, isometric force and Vmax were decreased by, respectively, about 15 and 25%. At 250 mM ionic strength, isometric force and Vmax were decreased by, respectively, 47 and 33%. 5. Vmax decreased with decreasing [Mg-ATP]. At [Mg-ATP] less than 0.1 mM there was no difference in Vmax between 35 and 150 mM ionic strength. At 250 mM ionic strength Vmax was lower than that at 150 mM at all [Mg-ATP]. 6. JATP during contraction in thiophosphorylated fibres at 35, 150 and 250 mM ionic strength was respectively, 0.62, 0.98 and 0.93 mumol g-1 min-1. The energetic tension cost (JATP/force) increased with ionic strength. 7. The force response to a quick stretch was investigated in the relaxed, contracted and rigor states at 25, 150 and 250 mM ionic strength. Stiffness in the relaxed state increased with speed of stretch and was higher the lower the ionic strength. In the contracted and rigor states, stiffness was also affected by ionic strength, but the relative effect in the contracted state was small.(ABSTRACT TRUNCATED AT 400 WORDS)


Osteogenic response to porous hydroxyapatite ceramics under the skin of dogs.
Soft tissue reactions to two materials of different microstructure made with porous and dense hydroxyapatite ceramic granules, were compared in this study. Each was implanted subcutaneously in the abdomen of 10 dogs. The implants were removed 1, 3 and 6 months after implantation, and examined by light microscopy. Heterotopic bone formation was only evident around porous hydroxyapatite ceramic granules in 6 of the 10 animals after 3 months, and in all cases after 6 months, but never around dense granules. These findings suggested that cellular differentiation and induction of osteogenesis may be influenced by the physical and chemical factors in the micromilieu provided by porous hydroxyapatite ceramic granules.


Consequences of noise- or styrene-induced cochlear damages on glutamate decarboxylase levels in the rat inferior colliculus.
Both noise and styrene can injure the cochlea, resulting in a reduction of incoming inputs from the cochlea to the central nervous system. In addition, styrene is known to have neurotoxic properties at high doses. The loss of inputs caused by noise has been shown to be compensated by a new equilibrium between excitatory and inhibitory influences within the inferior colliculus (IC). The main goal of this study was to determine whether styrene-induced hearing loss could also be counterbalanced by a GABAergic adjustment in the IC. For this purpose, rats were exposed to noise (97 dB SPL octave band noise centered at 8 kHz), or to a non-neurotoxic dose of styrene for 4 weeks (700 ppm, 6 h/day, 5 days/week). Auditory sensitivity was tested by evoked potentials, and cochlear damage was assessed by hair cell counts. Glutamate decarboxylase (GAD) was dosed in the IC by indirect competitive enzyme-linked immunosorbent assay. Both noise and styrene caused PTSs that reached 27.0 and 14.6 dB respectively. Outer hair cell (OHC) loss caused by noise did not exceed 9% in the first row, on the other hand OHC loss induced by styrene reached 63% in the third row. Only the noise caused a decrease of GAD of 37% compared to that measured in the controls. No significant modification of GAD concentration has been shown after styrene exposure. Thus, central compensation for cochlear damage may depend on the nature of the ototoxic agent. Unless styrene directly affects IC function, it is reasonable to assume that noise causes a modification of inhibitory neurotransmission within the structure because of impairment of afferent supply to the auditory brainstem. The present findings suggest that central compensation for cochlear damage can preferably occur when afferent fibers are altered.