Equal overall rejection rate in pre-transplant flow-cytometric cross-match negative and positive adult recipients in liver transplantation.
T cell IgG flow-cytometric cross-matches (FCXM) using 48 stored pre-transplant patient serum samples and 40 stored serum samples collected 3 wk after liver transplantation and frozen spleen cells of cadaveric donors in 48 consecutive liver transplantations were performed retrospectively. T cell IgG FCXM using pre-transplant serum samples was compared with 46 complement-dependent lymphocytotoxic cross-matches (CDCXM) performed at the time of transplantation. Clinical relevance of these tests was evaluated in relation to acute rejection, 1-, 3- and 5-yr graft and patient survival. The incidence of positive FCXM was 33% (16 of 48) and 13% (six of 46) by CDCXM. The median time of acute rejection was 29 d after transplantation in FCXM positive group (range 13-101 d) and 22 d in FCXM negative group (range 7-157 d, NS). Rejection rate was similar in 16 pre-transplant FCXM positive patients (eight of 16, 50%) compared with six pre-transplant CDCXM positive patients (three of six, 50%; NS). Recipients having graft rejection tended to be more often pre-transplant FCXM positive (eight of 21, 38%) than CDCXM positive (three of 21, 14%), but the difference was not significant (p > 0.1). No difference was found in the positive predictive value in relation to acute rejection between positive FCXM and CDCXM (69% vs. 50%; NS). Furthermore there was no correlation between post-transplant positive FCXM and acute rejection. No difference was found between pre-transplant T cell IgG FCXM positive and negative recipients in relation to graft or patient survival. Our findings are supportive for little risk associated with preformed donor-specific antibodies in liver transplantation.


Elevation of D-dimer levels are associated with early need for mechanical ventilation support in patients with COVID-19.
Severe COVID-19 disease is typically associated with an urgent need for supplemental oxygen therapy that may be successfully delivered through conventional methods or require invasive mechanical ventilation. Early prediction of the need for invasive mechanical ventilation could significantly improve outcomes of COVID-19 patients. Plasma levels of D-dimer and a number of inflammatory markers as well as values of complete blood counts, all measured in the first two days of hospital admission of COVID-19 patients, were evaluated for their significance as predictors of the eventual need for invasive mechanical ventilation support as well as their values as predictors of post-ventilation morbidly and mortality. This retrospective cohort study was conducted at a single center and included data pertaining to 200 patients with previously confirmed moderate to severe COVID-19 disease in the period between May 2021 and the end of December 2022. Data were retrieved from medical records for further analysis. The mean (SD) age of patients stood at 59 (14) years of age, and with a majority of patients being male (77%). About 18% of cases, all of significantly older age, had been connected to invasive mechanical ventilation (IMV). Total leucocytic count (TLC), as well as levels of urea, creatinine, D-dimer, ferritin, and CRP in IMV patients were significantly higher than non-ventilated patients (p < 0.01 for all). In contrast, lymphocytic count, hemoglobin level, and platelet count were significantly lower in IMV patients (p < 0.001, 0.04, and 0.002, respectively). The mortality rate was significantly higher in IMV patients (p < 0.001). D-dimer independently predicted IMV demand (OR = 1, p = 0.001 in adjusted and unadjusted models). The utility of D-dimer was excellent; and the cutoff level of above 1415 µ/L showed sensitivity and specificity of about 92% and 76%, respectively. Also, the D-dimer level was very effective in predicting post-IMV survival; the AUC = 0.86, p = 0.02, and a cutoff value below 4558 µ/L was associated with 100% and 66% sensitivity and specificity, respectively. High D-dimer levels independently correlated with the need for invasive mechanical ventilation. Low levels of this marker could evidently predict post-IMV survival of mechanically ventilated COVID-19 patients. Measuring D-dimer levels during routine follow up of those patients would thus be useful in predicting patient outcomes.


Magnetic covalent organic frameworks for extraction and determination of endocrine-disrupting chemicals in beverage and water samples.
Phenolic endocrine-disrupting chemicals (EDCs) are widespread and easily ingested through the food chain. They pose a serious threat to human health. Magnetic solid-phase extraction (MSPE) is an effective sample pre-treatment technology to determine traces of phenolic EDCs. Magnetic covalent organic framework (COF) (Fe3 O4 @COF) nanospheres were prepared and characterized. The efficient and selective extraction of phenolic EDCs relies on a large specific surface and the inherent porosity of COFs and hydrogen bonding, π-π, and hydrophobic interactions between COF shells and phenolic EDCs. Under optimal conditions, the proposed magnetic solid-phase extraction-high-performance liquid chromatography-ultra violet (MSPE-HPLC-UV) based on the metallic covalent organic framework method for phenolic EDCs shows good linearities (0.002-6 μg mL-1 ), with R2 of 0.995 or higher, and low limits of detection (6-1.200 ng mL-1 ). Magnetic covalent organic frameworks (Fe3 O4 @COFs) with good MSPE performance for phenolic EDCs were synthesized by the solvothermal method. The magnetic covalent organic framework-based MSPE-HPLC-UV method was applied successfully to determine phenolic EDCs in beverage and water samples with satisfactory recoveries (90.200%-123%) and relative standard deviations (2.100%-12.100%). © 2023 Society of Chemical Industry.


Reactivity of IgE and IgG serum levels to chymopapain after chemonucleolysis.
Patients suffering from herniated lumbar intervertebral disks can currently be treated by chemonucleolysis. This procedure involves reducing the nucleus pulposus by injecting chymopapain into the affected intervertebral disk. A small percentage of the patients who undergo this treatment experience an immediate hypersensitivity reaction. In this study the IgE and IgG specific for sensitivity to chymopapain of 21 patients receiving chymopapain (Discase for injection) was monitored over a 6-month period after injection. This study demonstrates that serum levels of IgE and IgG to chymopapain increase after chemonucleolysis.


The Role and Therapeutic Potential of miRNAs in Colorectal Liver Metastasis.
Colorectal cancer (CRC) is the fourth leading cause of cancer-related deaths worldwide. Liver metastasis is the major cause of CRC patient mortality, occurring in 60% patients with no effective therapies. Although studies have indicated the role of miRNAs in CRC, an in-depth miRNA expression analysis is essential to identify clinically relevant miRNAs and understand their potential in targeting liver metastasis. Here we analyzed miRNA expressions in 405 patient tumors from publicly available colorectal cancer genome sequencing project database. Our analyses showed miR-132, miR-378f, miR-605 and miR-1976 to be the most significantly downregulated miRNAs in primary and CRC liver metastatic tissues, and CRC cell lines. Observations in CRC cell lines indicated that ectopic expressions of miR-378f, -605 and -1976 suppress CRC cell proliferation, anchorage independent growth, metastatic potential, and enhance apoptosis. Consistently, CRC patients with higher miR-378f and miR-1976 levels exhibited better survival. Together, our data suggests an anti-tumorigenic role of these miRNAs in CRC and warrant future in vivo evaluation of the molecules for developing biomarkers or novel therapeutic strategies.


Activity staining of halophilic enzymes: substitution of salt with a zwitterion in non-denaturing electrophoresis.
Several NAD(P)(+)-dependent dehydrogenases were partially purified from Halobacterium halobium. When salt (2M (NH4)2SO4) was replaced with glycine betaine (4M or 6M), the zwitterion stabilised activity less completely than the salt. Nevertheless most of the enzyme activity still remained after 90h, e.g. 70% for malate dehydrogenase. This level of stabilisation permitted non-denaturing gel electrophoresis in 4M betaine after dialysis to replace salt. Coomassie Blue staining showed good separation of the proteins, and activity staining, hitherto impossible for halophilic enzymes, readily identified the individual dehydrogenase bands. Transfer of activity-stained gels to Coomassie staining solution halted background formazan staining and showed up activity and other protein bands in contrasting colours.


A biorefinery approach for pectin extraction and second-generation bioethanol production from cocoa pod husk.
A biorefinery approach was applied for pectin extraction, xylooligosaccharides' (XOs) and bioethanol production from cocoa pod husk (CPH) using citric acid-assisted hydrothermal pretreatment. Under optimal conditions at 120° C, 10 min and 2% w.v-1 of citric acid a high pectin recovery (19.5%) with high content of uronic acids (41.9%) was obtained. In addition, the liquid fraction presented a XOs concentration of 50.4 mg.g-1 and 69.7 mg.g-1 of fermentable sugars. Enzymatic hydrolysis of solid fraction showed glucan conversion of 60%. Finally, the hydrothermal and enzymatic hydrolysates of CPH were used in bioethanol production by Candida tropicalis and Saccharomyces cerevisiae, reaching 30.9 g and 45.2 g of bioethanol per kg of CPH, respectively. An environmentally friendly and rapid pretreatment method was development for pectin extraction, XOS and second-generation bioethanol production from CPH with great perspectives for the application of these biomolecules in food and bioenergy industry.


Driving after cerebral ischemia--a driving simulator investigation.
Stroke patients with relevant deficits are not allowed to drive. This study tended to assess driving abilities in an early stage after cerebral ischemia with a driving simulator. 32 patients (aged 54.9 +/- 14.8 years) with acute ischemia in the middle cerebral artery (MCA) (n = 24) and in the vertebral artery (VA) (n = 8) were investigated with reaction tests and a driving simulator. Controls were 12 healthy volunteers (aged 50.0 +/- 13.9 years). Compared to controls, the accident rate was increased only in patients with MCA-ischemia (2.9 +/- 3.6 vs. 1.3 +/- 1.4, p < 0.05). These patients also had poorer results in the driving simulator (p < 0.05) than patients with VA-ischemia. Driving simulator performance and neuropsychological test results did not correlate. All patients had only mild deficits and stated themselves able to drive. We could demonstrate the need to focus on driving skills especially in patients with MCA-infarctions. The driving simulator assesses various physical and neuropsychological functions influencing driving. It may especially be used for expert opinions regarding driving licence. It can also serve as a pre-driver evaluation before on road driving tests.


Coexistence of three species of leaf-feeding aphids (Homoptera) onBetula pendula.
Three sympatric aphid species of European origin (Euceraphis betulae (Koch),Callipterinella calliptera (Hartig) andBetulaphis brevipilosa Börner) feed onBetula pendula Roth in northern California. The component species of this guild partition resources by plant part, feeding sites within leaves and leaf phenological state. Overall niche overlap between species reached a maximum of 49.9% betweenB. brevipilosa andC. calliptera. Co-occurence ofE. betulae with both other species on leaves was random while co-occurrence ofC. calliptera andB. brevipilosa was aggregated. Morphological features (stylet length and body weight) act to mediate niche organization within this guild.California populations ofE. betulae andB. brevipilosa appear to utilize resources in a fashion similar to endemic Danish populations. Whereas DanishC. calliptera usually feed inside leaves silked together by spiders or Lepidoptera,C. calliptera in California also commonly feed on exposed leaf surfaces. This species, which has thus expanded its endemic niche, occupies the most generalized niche of the three species studied.


Differences in future life expectancy of testicular germ-cell tumor patients vs. age-matched male population-based controls.
It is unknown whether five-year overall survival (OS) differs and to what extent between testicular germ-cell tumor (TGCT) patients and age-matched male population-based controls. We identified newly diagnosed (2004-2014) TGCT patients within Surveillance Epidemiology and End Results database 2004-2019. We compared OS between non-seminoma (NS-TGCT) and seminoma (S-TGCT) patients relative to age-matched male population-based controls based on Social Security Administration Life-Tables. Smoothed cumulative incidence plots displayed cancer-specific mortality (CSM) vs. other-cause mortality (OCM). Of all 20,935 TGCT patients, 43% had NS-TGCT and 57% had S-TGCT. Of NS-TGCT patients, 63% were stage I vs. 16% stage II vs. 21% stage III. Of S-TGCT patients, 86% were stage I vs. 8% were stage II vs. 6% stage III. Five-year OS differences between NS-TGCT patients vs age-matched male population-based controls were 97 vs. 99% (Δ = 2%) for stage I, 96 vs. 99% (Δ = 3%) for stage II, 76 vs 98% (Δ = 22%) for stage III. Five-year OS differences between S-TGCT patients vs age-matched male population-based controls were 97 vs. 98% (Δ = 1%) for stage I, 95 vs. 97% (Δ = 2%) for stage II, 87 vs. 98% (Δ = 11%) for stage III. OCM rates ranged from 1 to 3% in NS-TGCT patients and from 2 to 4% in S-TGCT patients. The OS difference between NS-TGCT patients vs. age-matched male population-based controls was invariably higher across all stages (2-22%) than for S-TGCT patients (1-11%). Reassuringly, OCM rates were marginal in stage I and stage II patients. Conversely, higher OCM rates were recorded in stage III patients.


Japanese health and safety information for overseas visitors: a randomized controlled trial.
International travel to Japan increased steadily until the coronavirus disease 2019 (COVID-19) outbreak. Although international travel was curtailed worldwide due to the pandemic, the number of overseas visitors to Japan should increase again after the restrictions are lifted. We assessed the effect of a five-minute digital game on the knowledge of health information and the level of satisfaction with educational health resources of overseas visitors to Japan. We conducted a randomized controlled trial among 1062 previous and potential visitors to Japan utilizing an internet portal. We recruited previous and potential visitors to Japan from the internet portal sites of the UK, the US and Australia. We randomly allocated participants to two groups: an intervention group that played an animated game and a control group that viewed an online animation. All participants answered a self-administered questionnaire online from March 16 to 19, 2021. We assessed visitors' levels of health knowledge and satisfaction using the CSQ-8. We analyzed the data with a t test and the difference in differences test. Our RCT followed the SPIRIT guidelines. Of the 1062 previous and potential visitors recruited via the three countries' internet portals (354 from each country), some had visited Japan previously (174 in the intervention group, 220 in the control group), while some were potential visitors to Japan (357 in the intervention group, 311 in the control group). Some had gathered health and safety information about Japan prior to this study (180 in the intervention group, 211 in the control group). Both groups improved their health information levels after the intervention. The level of satisfaction with health information in Japan was significantly increased in the intervention group (average difference of 4.5 points) compared to that in the control group (average difference of 3.9 points) (p < 0.05). Both groups' mean CSQ-8 scores increased significantly after the intervention (p < 0.001): from 23 to 28 in the intervention group and from 23 to 24 in the control group. Our study introduced unique educational strategies using an online game to provide health and safety information to previous and potential visitors to Japan. The online game was a more effective way to increase satisfaction than the online animation about health information. This study was registered in the UMIN-CTR (University Hospital Medical Information Network Center Clinical Trials Registry) as Version 1, and the trial registration data are available as UMIN000042483, 17/11/2020. Trials UMIN-CTR (University Hospital Medical Information Network Center Clinical Trials Registry), UMIN000042483 (Japanese health and safety information for overseas visitors: A randomized controlled trial), 17/11/2020.


Localization of the ASV src gene product to the plasma membrane of transformed cells by electron microscopic immunocytochemistry.
The cellular location of the src gene product (p60src) of the Schmidt-Ruppin strain of avian sarcoma virus has been determined by electron microscopic immunocytochemistry in Schmidt-Ruppin ASV-transformed NRK cells, and the amount of the protein in different regions of the cell has been quantified. The protein is concentrated on the inner surface of the plasma membrane, particularly under ruffles, and it is highly concentrated on the inner surface of the membrane near junctions connecting adjacent cells. Small amounts of p60src were detected in the cytoplasm and in the perinuclear Golgi region of the cell. No significant localization was detected in control NRK cells or in NRK cells transformed by the Kirsten strain of murine sarcoma virus. The presence of p60src on the inner surface of the plasma membrane indicates that the changes in cell growth, cell shape and cell membrane structure noted in ASV-transformed cells are due to an initial action of p60src at the cell membrane.


[Xanthic lithiasis in the child. A study of one case. Review of the literature (author's transl)].
Seeking medical help because of an acute urinary retention, a 3-year-old child showed at intravenous urography (IVU) multiple radiotransparent lithiases. Hypouricemia suggested a diagnosis of xanthuria. The problems of this condition due to a congenital deficiency in xanthine oxydase are studied in a review of the literature. 56 xanthic lithiases have been published and since the work of Dent and Philpot, 28 xanthinurias have been reported.


Regulatory T cells in kidney disease and transplantation.
Regulatory T cells (Tregs) have been shown to be important in maintaining immune homeostasis and preventing autoimmune disease, including autoimmune kidney disease. It is also likely that they play a role in limiting kidney transplant rejection and potentially in promoting transplant tolerance. Although other subsets of Tregs exist, the most potent and well-defined Tregs are the Foxp3 expressing CD4(+) Tregs derived from the thymus or generated peripherally. These CD4(+)Foxp3(+) Tregs limit autoimmune renal disease in animal models, especially chronic kidney disease, and kidney transplantation. Furthermore, other subsets of Tregs, including CD8 Tregs, may play a role in immunosuppression in kidney disease. The development and protective mechanisms of Tregs in kidney disease and kidney transplantation involve multiple mechanisms of suppression. Here we review the development and function of CD4(+)Foxp3(+) Tregs. We discuss the specific application of Tregs as a therapeutic strategy to prevent kidney disease and to limit kidney transplant rejection and detail clinical trials in this area of transplantation.


Does abnormal neuronal excitability exist in myotonic dystrophy? I. Effects of the antiarrhythmic drug hydroquinidine on slow saccadic eye movements.
The abnormal neuronal excitability hypothesized in myotonic dystrophy (MD) might contribute to psychomotor and behavioral disturbances of MD patients. To gain new insights into the pathophysiology of MD, we determined whether the antiarrhythmic drug hydroquinidine would ameliorate slow saccadic eye movements (SEMs), apathy and hypersomnia in MD patients. SEMs were selected as simple modality for psychomotor investigation. The study was conducted in a randomized, placebo-controlled, double-blind, crossover manner. Ten ambulatory patients without contraindications to hydroquinidine administration were enrolled. Hydroquinidine (450 mg/day) or placebo was given orally for 6 weeks with a washout period of 6 weeks between treatments. SEMs were recorded by electrooculography and analyzed by a computer system. Two patients withdrew in the first week of active treatment because of nausea and epigastralgia. Hydroquinidine significantly increased the normalized peak saccadic velocity and shortened the saccadic reaction time compared to placebo. The drug's effects on apathy and hypersomnia are presented in a companion paper.


CELSR3 variants are associated with febrile seizures and epilepsy with antecedent febrile seizures.
To identify novel pathogenic gene of febrile seizures (FS)/epilepsy with antecedent FS (EFS+). The trio-based whole-exome sequencing was performed in a cohort of 462 cases with FS/EFS+. Silico programs, sequence alignment, and protein modeling were used to predict the damaging of variants. Statistical testing was performed to analyze gene-based burden of variants. Five heterozygous missense variants in CELSR3 were detected in five cases (families) with eight individuals (five females, three males) affected. Two variants were de novo, and three were identified in families with more than one individual affected. All the variants were predicted to be damaging in silico tools. Protein modeling showed that the variants resulted in disappearance of multiple hydrogen bonds and one disulfide bond, which potentially caused functional impairments of protein. The frequency of CELSR3 variants identified in this study was significantly higher than that in controls. All affected individuals were diagnosed with FS/EFS+, including six patients with FS and two patients with EFS+. All cases presented favorable outcomes without neurodevelopmental disorders. CELSR3 variants are potentially associated with FS/EFS+.


The cell structure damage and embden-meyerhof-parnas pathway inhibition of Listeria monocytogenes induced by glycinin basic peptide.
Glycinin basic peptide (GBP) is a natural antibacterial peptide. This study aimed to explore the antibacterial characteristics of GBP against Listeria monocytogenes (L. monocytogenes) by measuring the membrane potential, membrane permeability, cell damage, morphological changes, respiration metabolism inhibition and DNA content. GBP increased the surface zeta potential and decreased the trans-membrane potential of L. monocytogenes in a dose-dependent manner. Compared with the control, the electrical conductivities of GBP-treated bacterial suspensions were significantly increased. The percentages of bacteria with damaged membrane increased from 6.40% to 70.90% with GBP from 0 to 0.8 mg/mL. Obvious rupture and deform of bacterial cells with GBP were observed by transmission electron microscope (TEM), showing the destructive effect of GBP on L. monocytogenes. GBP also inhibited the embden-meyerhof-parnas pathway of the bacterial respiration metabolism and reduced the activities of its key regulator enzymes. Besides, the content of DNA in GBP-treated L. monocytogenes was lower than that in control.


Crystal structure of the guanylate kinase domain from discs large homolog 1 (DLG1/SAP97).
Discs large homolog 1 (DLG1/SAP97) is involved in the development and regulation of neuronal and immunological synapses. DLG1 is a member of the membrane associated guanylate kinase (MAGUK) family of proteins, which function as molecular scaffolds. The C-terminal guanylate kinase (GK) domain of DLG1 binds peptides with a phosphorylated serine residue. In this study, we solved the crystal structure of the GK domain of human DLG1. The C-terminal tail of DLG1 is bound to the peptide-binding site of an adjacent symmetry-related DLG1 GK molecule. The binding direction of the C-terminal tail to the peptide-binding site is opposite to that of the phosphorylated LGN peptide in complex with the rat DLG1 GK domain. The C-terminal tail forms a 310 helix, which is also different from the conformation of the phosphorylated LGN peptide. Nevertheless, the side chain interactions of the C-terminal tail with the DLG1 GK domain are similar to those of the phosphorylated LGN peptide.


Glioblastoma stem cells and Wnt signaling pathway: molecular mechanisms and therapeutic targets.
Glioblastoma is the most common form of primary brain tumor. Glioblastoma stem cells play an important role in tumor formation by activation of several signaling pathways. Wnt signaling pathway is one such important pathway which helps cellular differentiation to promote tumor formation in the brain. Glioblastoma remains to be a highly destructive type of tumor despite availability of treatment strategies like surgery, chemotherapy, and radiation. Advances in the field of cancer biology have revolutionized therapy by allowing targeting of tumor-specific molecular deregulation. In this review, we discuss about the significance of glioblastoma stem cells in cancer progression through Wnt signaling pathway and highlight the clinical targets being potentially considered for therapy in glioblastoma.


Time to completion of web-based physics problems with tutoring.
We studied students performing a complex learning task, that of solving multipart physics problems with interactive tutoring on the web. We extracted the rate of completion and fraction completed as a function of time on task by retrospectively analyzing the log of student-tutor interactions. There was a spontaneous division of students into three groups, the central (and largest) group (about 65% of the students) being those who solved the problem in real time after multiple interactions with the tutorial program (primarily receiving feedback to submitted wrong answers and requesting hints). This group displayed a sigmoidal fraction-completed curve as a function of logarithmic time. The sigmoidal shape is qualitatively flatter for problems that do not include hints and wrong-answer responses. We argue that the group of students who respond quickly (about 10% of the students) is obtaining the answer from some outside source. The third group (about 25% of the students) represents those who interrupt their solution, presumably to work offline or to obtain outside help.


Modelling sensitivity losses in ocular disorders: colour vision anomalies following intense blue-light exposure in monkeys.
The effects of prolonged exposure to intense, short-wavelength light were studied in monkeys through the measurement of increment-threshold spectral sensitivity (ITSS) and threshold-versus-intensity (TVI) functions using a behavioural method. The long-term effect of intense blue-light exposure was to induce a short-wavelength (SW) sensitivity loss which did not depend on the intensity or chromatic composition of the adapting field. The TVI curves for short wavelength stimuli revealed an increase in test threshold without changes in field sensitivity. Since this SW sensitivity loss may generalize to characteristic colour vision defects found in many outer retinal diseases, models of acquired alterations of colour vision mechanisms are considered. These models describe probable changes in ITSS functions and TVI curves in diseases affecting the inner or outer retina as well as changes in dark adaptation.


Giant multilocular epidermoid cyst on the left buttock.
Epidermoid cysts are the most common cysts of the skin. They are generally small and slow-growing and rarely reach more than 5 cm in diameter. We present a patient with a giant multilocular epidermoid cyst. A 75-year-old man presented with a giant, soft, painless, tumorlike mass on the left buttock that had gradually enlarged over a 12-year period. Excision of the mass from the surrounding tissue was extremely easy because of a well-defined capsule. The 28 x 14 x 12 cm mass was totally excised. A histopathologic examination revealed that it was an epidermoid cyst. This rare case of giant epidermoid cyst was treated successfully by local excision followed by primary closure. Dissection of the mass from the surrounding tissue was extremely easy because of a well-defined capsule.


Investigation of thermodynamic parameters in the pyrolysis conversion of biomass and manure to biochars using thermogravimetric analysis.
The thermodynamic parameters of the conversion of two companion pair materials, i.e., rice straw vs dairy manure, and rice bran vs chicken manure, to biochars were characterized by thermogravimetric analysis. The overall changes of activation energy (Ea) were well described by the Flynn-Wall method. The Ea values increased steeply from about 120 to 180 kJ/mol at the mass conversion (α) at 0.2-0.4, followed by a relatively steady change at 0.4<α<0.65, thereafter showed a quick increase at α>0.65. The higher contents of minerals in manures resulted in the larger Ea. The individual conversion of hemicellulose, cellulose and lignin in the feedstocks was identified and their thermodynamic parameters (ΔH°, ΔG° and ΔS°) were calculated. The yields of biochars calculated from TG curve were compared with the determined yields of biochars using muffle pyrolysis. Along with Fourier transform infrared spectra data, the distinct decompositions of biomasses and manures were evaluated.


Flow velocity waveforms in the maternal uteroplacental and fetal umbilical placental circulations.
A simple continuous-wave Doppler ultrasound system was used to record arterial flow velocity time waveforms from branches of the maternal uterine artery in the placental bed and the fetal umbilical arteries. The systolic-diastolic ratio of flow velocities was measured as an index of peripheral resistance. In normal pregnancy both circulations exhibit high diastolic flow velocities caused by low resistance. The study group consisted of 172 pregnancies with high fetal risk of which 53 resulted in delivery of a small-for-gestational age infant. The last study-to-delivery interval was less than 10 days. In the small-for-gestational age fetuses in whom both umbilical and uterine studies were normal there was good fetal outcome (19 cases). Neonatal morbidity occurred among those with abnormal umbilical studies with low diastolic flow velocities, indicating high resistance. Of these studies there were two subgroups. An abnormal uterine study (13 cases) indicated a primary maternal uteroplacental lesion and a normal study (21 cases) a primary fetal lesion. In cases of severe maternal hypertension, abnormal uterine artery waveforms were associated with abnormal umbilical artery waveforms.


Motivations for prescription drug misuse among young men who have sex with men (YMSM) in Philadelphia.
Prescription drug misuse (i.e. opioids, tranquilizers and stimulants) has become the fastest growing area of substance abuse among young adults. Limited studies focus on prescription drug misuse among young men who have sex with men (YMSM, aged 18-29 years). Furthermore, little is known about YMSM's motivations for misuse. The purpose of this study was to explore personal motivations for prescription drug misuse among YMSM, including the possible connection between misuse and sexual behaviors. As part of a larger mixed methods study of 191 YMSM recruited in Philadelphia during 2012-2013, we conducted semi-structured qualitative interviews with 25 of these participants to gather additional contextual information about their prescription drug misuse. We conducted thematic analysis of qualitative data. While our results corroborated previous literature on motives for misuse of prescription drugs, our data yielded some distinct motivations specific among YMSM. These motives included social/recreational motives, facilitating sex with other men (including motives such as use of opioids for less painful anal receptive sex), and psychological motives such as depression, stress management, coping with everyday hardships (opioids and tranquilizers) or feeling more energized (stimulants). Prescription drugs were commonly misused within the broader contexts of participants' polysubstance use, adding to the significance of this problem. Our findings offer insights into YMSM's motivations for prescription drug misuse, and point to the importance of recognizing and addressing them. While substance use is likely related to various psychosocial issues impacting YMSM, it also may lead to significant health consequences. Results support the need to include prescription drugs and polysubstance use in harm reduction messages and treatment approaches aimed at substance using YMSM.


Topical ROR Inverse Agonists Suppress Inflammation in Mouse Models of Atopic Dermatitis and Acute Irritant Dermatitis.
The retinoic acid receptor-related orphan receptors RORα and RORγ are critical for the functions of specific subsets of T cells and innate lymphoid cells, which are key drivers of inflammatory disease in barrier tissues. Here, we investigate the anti-inflammatory potential of SR1001, a synthetic RORα/γ inverse agonist, in mouse models of atopic dermatitis and acute irritant dermatitis. Topical treatment with SR1001 reduces epidermal and dermal features of MC903-induced atopic dermatitis-like disease and suppresses the production of type 2 cytokines and other inflammatory mediators in lesional skin. In the epidermis, SR1001 treatment blocks MC903-induced expression of TSLP and reverses impaired keratinocyte differentiation. SR1001 is also effective in alleviating acute dermatitis triggered by 12-O-tetradecanoylphorbol-13-acetate. Overall, our results suggest that RORα/γ are important therapeutic targets for cutaneous inflammation and suggest topical usage of inhibitory ligands as an approach to treating skin diseases of inflammatory etiology.


Characterization of an angiotensin II-fluorescamine derivative.
The coupling of fluorescamine (4-phenylspiro[furan-2(3H), 1'-PHTHALAN]-3,3'dione) to angiotensin II to form a fluorescent derivative was studied. Complete reaction of the peptide below concentrations of 10- minus 4 M could be achieved with a fluorescamine concentration of 0-3 mg ml- minus 1 of acetone at pH 8-3, and the lowest concentration detectable by fluorescence spectroscopy was 100 pmol ml- minus 1. The derivative, as prepared did not react with ninhydrin, and no fluorescence was generated when fluorescamine was reacted with (1-Sar)-ATII. These data suggest that fluorescence is generated only through the coupling of fluorescamine to the N-terminal primary amine of ATII. The ATII-fluorescamine derivative has the same intrinsic activity on the contraction of rat colon (elevenfold loss of affinity), and on the release of fluorogenic corticosteroids from bovine adrenal cortical slices (sixfold loss of affinity) compared to ATII. Water-hydrolysed fluorescamine and Asp-fluorescamine did not contract rat colon preparations; the contractile response to ATII-fluorescamine was blocked by (8-Leu)-ATII, a specific ATII antagonist. These findings suggest that theATII fluorophore shares a common receptor site with the native octapeptide. The rate loss of biological activity of the ATII-fluorescamine derivative was appreciably lower than that observed for ATII. The present study suggests that the ATII-fluorescamine derivative can be substituted for radioactively-labelled ATII for use in a variety of applications.


Effect of microwave pre-treatment on quality parameters in Indian mustard.
Demand for edible oil from crops like groundnut, mustard and soybean, is increasing and likely to increase further. In this regard, Indian mustard seeds were treated with microwaves (MW), to investigate the possibility of enhancing oil yield and nutritional content. MW pre-treatment was given to the seeds of two Indian mustard varieties i.e. PM21 (V1) and PDZ1 (V2) for 0, 2, 4 and 6 min (H0, H2, H4 and H6 min respectively). MW treatment with increasing exposure time showed a linear reduction in the glucosinolate and erucic acid content with concomitant increase in oil yield and moisture loss in both the varieties, as evident from correlation and principal component analysis. Antioxidant capacity has increased with the reduction in phytic acid content (1.82) in V2 at 6 min exposure time with respect to untreated control. Free radical scavenging activity was improved with increasing MW treatment in both the varieties. Therefore, from the obtained results, it is advisable to treat mustard seeds with MW before extraction of oil, because it gives a relatively good oil yield, with enhanced nutritional factors. Moreover, microwaving was effective in reducing glucosinolates and erucic acid also.


Nerve conduction studies and EMG in carpal tunnel syndrome: Do they add value?
This paper summarises the views of four experts on the place of neurophysiological testing (EDX) in patients presenting with possible carpal tunnel syndrome, in guiding their treatment, and in reevaluations. This is not meant to be a position paper or a literature review, and heterogeneous viewpoints are presented. Nerve conduction studies should be performed in patients presenting with possible carpal tunnel syndrome to assist diagnosis, and may need to be repeated at intervals in those managed conservatively. There is evidence that local corticosteroid injection is safe and effective for many patients, thereby avoiding or deferring surgical decompression. All patients should undergo EDX studies before any invasive procedure for CTS (injection or surgery). Needle EMG studies are not obligatory, but may be needed in those with severe disease and those in whom an alternate or concomitant diagnosis is suspected.


Direction and distance dependency of reaching movements of lower limb.
Efficient body movement is required in our daily lives, as it facilitates responding to the external environment and producing movements in various directions and distances. While numerous studies have reported on goal-directed movements in the frontal direction during gait initiation, there is limited research on the efficient movement of the lower limbs in multiple directions and distances. Therefore, we aimed to examine changes in the kinematics of lower-limb reaching movements to determine skilled motor ability in terms of direction and distance. Sixteen adults (10 male participants) were requested to reach targets projected on the floor in seven directions and at three distances for a total of 21 points. The reaching time slowed down for the contralateral side (right foot to left-sided target) and was caused by a slower start of the toe movement. To identify the cause of this delay, we analyzed the onset of movement at each joint and found that movement to the contralateral side starts from the hip, followed by the knee, and subsequently the toe. The time-to-peak velocity was also calculated, and the motion required to reach the target in the shortest time varied depending on direction and distance. These results suggested that movement kinematics vary with direction and distance, resulting in a slower reaching time on the contralateral side. The results of our study hold promise for potential applications in sports and rehabilitation.


The effect of an aldose reductase inhibitor on cardiovascular performance in patients with diabetes mellitus.
Because some aldose reductase inhibitor studies have demonstrated clinical improvement in scored neurological signs and symptoms of diabetic neuropathy, a prospective study of the effect on cardiovascular performance of sorbinil 250 mg/day for 12 months was conducted on patients with diabetic autonomic neuropathy who were free of atherosclerotic coronary artery disease and/or cardiomyopathy. After 1 year of treatment, the study group (n = 14) demonstrated significant improvement in both the resting cardiac output (P = 0.02), and the maximal cardiac output (P = 0.03). This observation suggests that the use of an aldose reductase inhibitor may be useful in treating suboptimal cardiovascular performance in patients with diabetic cardiac autonomic neuropathy.


Toxic and transcriptional responses of PC12 cells to soluble tungsten alloy surrogates.
There is increasing evidence that metals have a role in the etiology of diverse neurological diseases. This study used PC12 cells as an in vitro model to examine the toxicity of tungsten alloys that have important military applications. Initially, the relative concentrations of tungsten (W), nickel (Ni), and cobalt (Co) mobilized from pellets of a weapons-grade tungsten alloy incubated in physiologically relevant solutions were determined. Dosing solutions of soluble metal salts that were equivalent in ratio to those mobilized from these alloy pellets were used to treat nerve growth factor (NGF) differentiated PC12 cells. Treatments consisted of single (W, Ni or Co), paired (W/Ni, W/Co or Ni/Co) or complete (W/Ni/Co) metal exposures for 24 h followed by measurement of cytotoxicity, viability, and microarray analysis to examine their impact on survival and viability, global gene expression, and biological processes. Gene expression changed dramatically with addition of NGF. Addition of Ni or Co either singly or in combination further impacted gene expression. An observed additive effect of Ni and Co on gene expression was unaffected by the addition of W. The work showed that tungsten, as found in this tungsten alloy, had minimal relative toxicity as compared to the other alloy components when used either alone or in combination.


Hypertrophic cardiomyopathy.
Hypertrophic cardiomyopathy is a fascinating disease of marked heterogeneity. Hypertrophic cardiomyopathy was originally characterized by massive myocardial hypertrophy in the absence of known etiology, a dynamic left ventricular outflow obstruction, and increased risk of sudden death. It is now well accepted that multiple mutations in genes encoding for the cardiac sarcomere are responsible for the disease. Complex morphologic and pathophysiologic differences, disparate natural history studies, and novel treatment strategies underscore the challenge to the practicing cardiologist when faced with the management of the hypertrophic cardiomyopathy patient.


Tranexamic acid for bleeding: Much more than a treatment for postpartum hemorrhage.
The evidence that early tranexamic acid treatment reduces postpartum hemorrhage deaths has major implications for obstetrical care worldwide. Tranexamic acid may also have a role in the prevention of postpartum hemorrhage, but more evidence is needed on the balance of risks and benefits. Most deaths from postpartum hemorrhage are in low- and middle-income countries where tranexamic acid treatment is often unavailable. Several maternal health organizations including the Reproductive Health Supplies Coalition, Clinton Health Access Initiative, Concept Foundation, International Federation of Gynecology and Obstetrics, and Unitaid are working to increase access. However, a wider view of the evidence on tranexamic acid and bleeding shows that it can improve maternal health in many other ways. An appreciation of these other health benefits could facilitate efforts to increase access. By reducing heavy menstrual bleeding, tranexamic acid could reduce the prevalence of maternal anemia, a common and important risk factor for postpartum hemorrhage and other maternal and neonatal outcomes. Further clinical trials of tranexamic acid for the treatment of menstrual bleeding are needed. By reducing surgical bleeding and the need for blood transfusion, tranexamic acid would increase the availability of blood in countries where there is blood shortage so that more blood is available for use in life-threatening bleeding including postpartum hemorrhage. In countries where there is no blood shortage, tranexamic acid use would reduce healthcare costs and prevent transfusion-transmitted infections and reactions. Trauma affects women and men, and violence is a leading cause of death in pregnancy. Increased use of tranexamic acid in trauma would significantly reduce trauma deaths. Efforts to increase the availability and use of tranexamic acid for obstetrical hemorrhage should acknowledge its other health benefits and aim to increase its use across health services more generally.


Fat graft survival requires metabolic reprogramming toward the glycolytic pathway.
Fat grafts are widely used as natural fillers in reconstructive and cosmetic surgery. However, the mechanisms underlying fat graft survival are poorly understood. Here, we performed an unbiased transcriptomic analysis in a mouse fat graft model to determine the molecular mechanism underlying free fat graft survival. We conducted RNA-sequencing (RNA-seq) analysis in a mouse free subcutaneous fat graft model on days 3 and 7 following grafting (n = 5). High-throughput sequencing was performed on paired-end reads using NovaSeq6000. The calculated transcripts per million (TPM) values were processed for principal component analysis (PCA), unsupervised hierarchically clustered heatmap generation, and gene set enrichment analysis. PCA and heatmap data revealed global differences in the transcriptomes of the fat graft model and the non-grafted control. The top meaningful upregulated gene sets in the fat graft model were related to the epithelial-mesenchymal transition, hypoxia on day 3, and angiogenesis on day 7. Mechanistically, the glycolytic pathway was upregulated in the fat graft model at days 3 (FDR q = 0.012) and 7 (FDR q = 0.084). In subsequent experiments, pharmacological inhibition of the glycolytic pathway in mouse fat grafts with 2-deoxy-D-glucose (2-DG) significantly suppressed fat graft retention rates, both grossly and microscopically (n = 5). Free adipose tissue grafts undergo metabolic reprogramming toward the glycolytic pathway. Future studies should examine whether targeting this pathway can enhance the graft survival rate.


NMR-based metabolic profiling to follow the production of anti-phytopathogenic compounds in the culture of the marine strain Streptomyces sp. PNM-9.
Actinobacteria are the major source of bioactive secondary metabolites and are featured in the search for antimicrobial compounds. We used nuclear magnetic resonance (RMN)-metabolic profiling and multivariate data analysis (MVDA) to correlate the metabolites' production of Streptomyces sp. PNM-9 from the algae Dictyota sp. and their biological activity against the rice phytopathogenic bacteria Burkholderia spp. The compounds 2-methyl-N-(2'-phenylethyl)-butanamide (1) and 3-methyl-N-(2'-phenylethyl)-butanamide (2) were identified through MVDA and 2D NMR experiments in the organic extract of a 15-days LB media culture of Streptomyces sp. PNM-9. Compounds 1 and 2 were isolated and their structures confirmed by one- and two-dimensional NMR and mass spectrometry (MS) data. Compounds 1 and 2 were active against the rice pathogenic bacteria Burkholderia glumae (ATCC 33,617) displaying minimal inhibitory concentration (MIC) values of 2.43 mM and 1.21 mM, respectively. The metabolomics-guided approach employing NMR-metabolic profiling was useful for marine microbial bioprospecting and suggested Streptomyces sp. PNM-9 strain and its compounds as a potential control against phytopathogenic bacteria.


Fetal alcohol syndrome in families.
A retrospective study of the clinical literature of fetal alcohol syndrome (FAS) was conducted to determine the incidence of FAS among younger and older siblings of a patient with FAS. The incidence of the disorder among older sibs was 170 per 1,000 whereas among younger sibs the incidence was 771 per 1,000. Since the risk for FAS in the general population is 1.9 per 1,000, these results clearly indicate the high risk of FAS among siblings if one sib is diagnosed as FAS.


Comparison and evolutionary analysis of the glycosomal glyceraldehyde-3-phosphate dehydrogenase from different Kinetoplastida.
In this work, we present the sequences and a comparison of the glycosomal GAPDHs from a number of Kinetoplastida. The complete gene sequences have been determined for some species (Crithidia fasciculata, Herpetomonas samuelpessoai, Leptomonas seymouri, and Phytomonas sp), whereas for other species (Trypanosoma brucei gambiense, Trypanosoma congolense, Trypanosoma vivax, and Leishmania major), only partial sequences have been obtained by PCR amplification. The structure of all available glycosomal GAPDH genes was analyzed in detail. Considerable variations were observed in both their nucleotide composition and their codon usage. The GC content varies between 64.4% in L. seymouri and 49.5% in the previously sequenced GAPDH gene from Trypanoplasma borreli. A highly biased codon usage was found in C. fasciculata, with only 34 triplets used, whereas in T. borreli 57 codons were employed. No obvious correlation could be observed between the codon usage and either the nucleotide composition or the level of gene expression. The glycosomal GAPDH is a very well-conserved enzyme. The maximal overall difference observed in the amino acid sequences is only 25%. Specific insertions and extensions are retained in all sequences. The residues involved in catalysis, substrate, and inorganic phosphate binding are fully conserved, whereas some variability is observed in the cofactor-binding pocket. The implications of these data for the design of new trypanocidal drugs targeted against GAPDH are discussed. All available gene and amino acid sequences of glycosomal GAPDHs were used for a phylogenetic analysis. The division of the Kinetoplastida into two suborders, Bodonina and Trypanosomatina, was well supported. Within the letter group, the Trypanosoma species appeared to be monophyletic, whereas the other trypanosomatids form a second clade.


Enhanced Proton Conductivity by Aliovalent Substitution of Cadmium for Indium in Dimethylaminium-Templated Metal Anilicates.
Metal-organic frameworks (MOFs) with excellent proton conducting ability are crucial to fuel cells, chemical sensors, and redox flow batteries, but achieving them remain a challenge because of the difficulty in simultaneous fulfillment of large number of proton carriers, high mobility of protons, and long-term durable proton conduction. To explore a simple, efficient, and general route toward highly proton-conducting MOFs, we propose herein an aliovalent substitution metal strategy for isostructural aminium-templated MOFs which benefit the acquisition of rich proton sources without modifying ligands or exchanging protic organic molecules. This idea is verified by 100-fold enhancement of conductivity in compounds (Me2NH2)2[Cd(mdhbqdc)2] (Cd-BQ) and (Me2NH2) (Me2NH)[In(mdhbqdc)2] (In-BQ) (H2mdhbqdc = dimethyl 3,6-dihydroxy-2,5-benzoquinone-1,4-dicarboxylic acid) that feature three-dimensional diamond-like structures with two-dimensional intersected channels. Accompanied by the in situ formation of an anilicate ligand, a great number of -OH groups are grafted onto the inner wall of pores, which interact with neutral Me2NH and/or protonated Me2NH2+ cations via N-H···O hydrogen bonds. The high concentration of protons and dynamics of protic amines in the porous framework readily leads to a moderate conductivity of In-BQ (2.10 × 10-4 S cm-1, at 303 K under 95% RH) and an activation energy of 0.73 eV (95% RH). It should be noted that the aliovalent substitution of Cd(II) for In(III) results in the doubling of dimethylaminium proton carriers in Cd-BQ, indicating more frequent hopping and multiple proton-transfer pathways. This indication is supported by a very high protonic conductivity of 2.30 × 10-2 S cm-1 and a reduced activation energy of 0.48 eV under the same conditions. Molecular dynamics simulations visually elucidate the fact that compared with In-BQ, aliovalent-substituted Cd-BQ has shorter proton-migration distances, which in combination with more proton numbers results in more frequent hopping and sliding of protons, in agreement with the experimental results.


Characterization of olive oil flavored with Brazilian pink pepper (Schinus terebinthifolius Raddi) in different maceration processes.
The present study aimed to evaluate two different processes of olive oil aromatization with Schinus terebinthifolius Raddi, conventional maceration (CM) and ultrasound-assisted maceration (UM), and their influence on quality parameters, total phenolic compounds (TPC), fatty acid profile (FA), volatile organic compounds (VOCs), antioxidant capacity, and oxidative stability. Flavoring reduced peroxide values, although it increased free fatty acids and extinction coefficients. The flavorization process did not change the FA profile, which showed oleic acid as a major compound. The VOCs varied and the migration of oxygenated monoterpenes were more effective in UM-flavored olive oil compared to its CM counterpart. All flavored olive oils presented higher oxidative stability than the control samples and UM-flavored olive oil was highlighted for its higher antioxidant activity. These findings proved that aromatization with pink pepper assisted by ultrasound may be an alternative to improve olive oil quality and create a new competitive flavored product.


Research topics in intellectual disabilities in North Korea: A scoping review.
Research outcomes on intellectual development and related disabilities in North Korea are not widely known. Therefore, the current scoping review aimed to provide preliminary insight on research topics concerning intellectual disabilities in North Korea. A six-stage framework for scoping review was adopted to examine research trends. Articles were categorised based on the era of supreme leader and research topic. There is a greater amount of research regarding intellectual disabilities in the recent Kim Jong-un era compared to the period of the previous leader where research outcomes on general intelligence were the focus. Significant qualitative progress was similarly found. The current analysis on research outcomes provides meaningful insights to aid in understanding the atmosphere in North Korea surrounding intellectual disabilities. Follow-up studies and open discussions are necessary for further progress.


To tell or not to tell: attitudes of transplant surgeons and transplant nephrologists regarding the disclosure of recipient information to living kidney donors.
Organ Procurement and Transplantation Network policy regarding "Living Donation Informed Consent Requirements" only requires general disclosure of candidate health information to prospective living kidney donors. We examined attitudes of transplant surgeons and transplant nephrologists regarding greater disclosure. Web-based and mailed surveys to explore attitudes about disclosing potential recipient health information, health-associated behaviors, and lifestyle choices to living donors. Of 397 potential participants, 111 eligible participants (28%) fully or partially responded. Respondents were split between surgeons (42%) and nephrologists (58%). While 72% believed that general disclosure did not require explicit permission, 88% believed that disclosure of specific recipient information did. Many would disclose more information if legally permissible. Over 65% thought disclosure of recipient information should not depend on the donor-recipient relationship. Virtually all supported disclosing expected one- and five-yr graft survival and anticipated deceased donor wait-time. Sixty-six percent supported disclosing non-compliance or difficulty taking medications. Support was divided for disclosure of HIV (52%), hepatitis (49%), smoking (53%), illicit drugs (50%), alcohol (49%), and psychiatric history (44%). While virtually all respondents support disclosing recipient information directly relevant to graft and patient survival to prospective living donors, they are divided about sharing other recipient health, health behavior, and/or lifestyle information.


Regulation of glucose and lipid metabolism in health and disease.
Glucose and fatty acids are the major sources of energy for human body. Cholesterol, the most abundant sterol in mammals, is a key component of cell membranes although it does not generate ATP. The metabolisms of glucose, fatty acids and cholesterol are often intertwined and regulated. For example, glucose can be converted to fatty acids and cholesterol through de novo lipid biosynthesis pathways. Excessive lipids are secreted in lipoproteins or stored in lipid droplets. The metabolites of glucose and lipids are dynamically transported intercellularly and intracellularly, and then converted to other molecules in specific compartments. The disorders of glucose and lipid metabolism result in severe diseases including cardiovascular disease, diabetes and fatty liver. This review summarizes the major metabolic aspects of glucose and lipid, and their regulations in the context of physiology and diseases.


A model to study the direct effect of diet on early intestinal growth and maturation using Thiry-Vella loops.
Thiry-Vella loops (TVL) are neurovascular intact intestinal segments isolated from the normal flow of chyme. TVL have been used to assess the effects of local and systemic factors in the intestine of adult animal models. To assess the direct effect of diet in growth and maturation of the intestine in the growing animal, the authors developed a similar model in the weanling rat. The surgical technique is described. Results include weight gain, morphometry of the intestine, dissacharidases, total proteins, and nucleic acid in both TVLs and intestine in continuity. This model is inexpensive, easy to handle, and ideal for studying the effect of dietary factors on intestinal growth and maturation, separating the direct effect of diet on the intestine from systemic effects.


Bone tissue in reflex sympathetic dystrophy syndrome--Sudeck's atrophy: structural and ultrastructural studies.
Light and electron microscopic examination of bone in reflex sympathetic dystrophy syndrome confirms that vascular disorders play an important role in the genesis of bone lesions. These vascular disorders lead to degeneration of osteocytes and demineralization of bone, with the disappearance of hydroxyapatite crystals. The neo-osteogenesis that follows resembles the different stages of fracture repair, with precocious elaboration of irregular, replacement woven bone, followed by osteoclastic resorption and the laying down of lamellar bone trabeculae. The bone lesions clearly appear to be a consequence of autonomic nervous system circulatory disorders.


Gadolinium chloride-induced shifts in intrahepatic distributions of liposomes.
Intravenously administered gadolinium chloride caused only a slight decrease in the rate of elimination of small unilamellar liposomes from the blood and had no influence on the total hepatic uptake of these vesicles, but did alter their intrahepatic distribution substantially. Uptake by the non-parenchymal cells was substantially decreased, whereas uptake by the parenchymal cells showed a concomitant increase. Our earlier observations (Roerdink et al. (1981) Biochim. Biophys. Acta 677, 79-89) on the effect of lanthanides on the in vivo distribution of multilamellar liposomes have been extended, in that we demonstrate, in addition to the drop in elimination rate from the blood and in the over-all hepatic uptake, a shift of liposome distribution within the Kupffer cell population. While the larger Kupffer cells, which normally take up a major fraction of an injected liposome dose, were strongly inhibited in liposome uptake, the more numerous small macrophages showed a 3-4-fold increase in uptake.


A 3-year prospective randomized controlled clinical trial of group care in type 1 diabetes.
We showed that continuing education can be embedded into routine diabetes care by seeing patients in small groups rather than individually. Group care was cost-effective in improving quality of life, knowledge of diabetes, health behaviours and clinical outcomes in people with type 2 diabetes. The aim of this study was to verify if group care can also be applied to type 1 diabetes. Randomized, controlled clinical trial comparing 31 patients managed by group care with 31 managed by traditional one-to-one care. A syllabus was built and later remodulated with the patients in a series of focus-group meetings. The primary end-point was changes in quality of life. Secondary end-points were: knowledge of diabetes, health behaviours, HbA1c and circulating lipids. Differential costs to the Italian National Health System and to the patients were also calculated. After 3 years, quality of life improved among patients on group care, along with knowledge and health behaviours (p<0.001, all). Knowledge added its effects to those of group care by independently influencing behaviours (p=0.004) while quality of life changed independently of either (p<0.001). Among controls, quality of life worsened (p<0.001) whereas knowledge and behaviours remained unchanged. HDL cholesterol increased among patients on group care (p=0.027) and total cholesterol decreased in the controls (p<0.05). HbA1c decreased, though not significantly, in both. Direct costs for group and one-to-one care were Euros 933.19 and Euros 697.10 per patient, respectively, giving a cost-effectiveness ratio of Euros 19.42 spent per point gained in the quality of life scale. Group care is applicable and also cost-effective in type 1 diabetes. It improves quality of life, knowledge and behaviours. Future programme adjustments should strive to impact more on metabolic control.


Production of high quality brain-derived neurotrophic factor (BDNF) and tropomyosin receptor kinase B (TrkB) RNA from isolated populations of rat spinal cord motor neurons obtained by Laser Capture Microdissection (LCM).
The mammalian central nervous system (CNS) is composed of multiple cellular elements, making it challenging to segregate one particular cell type to study their gene expression profile. For instance, as motor neurons represent only 5-10% of the total cell population of the spinal cord, meaningful transcriptional analysis on these neurons is almost impossible to achieve from homogenized spinal cord tissue. A major challenge faced by scientists is to obtain good quality RNA from small amounts of starting material. In this paper, we used Laser Capture Microdissection (LCM) techniques to identify and isolate spinal cord motor neurons. The present analysis revealed that perfusion with paraformaldehyde (PFA) does not alter RNA quality. RNA integrity numbers (RINs) of tissue samples from rubrospinal tract (RST)-transected, intact spinal cord or from whole spinal cord homogenate were all above 8, which indicates intact, high-quality RNA. Levels of mRNA for brain-derived neurotrophic factor (BDNF) or for its tropomyosin receptor kinase B (TrkB) were not affected by rubrospinal tract (RST) transection, a surgical procedure that deprive motor neurons from one of their main supraspinal input. The isolation of pure populations of neurons with LCM techniques allows for robust transcriptional characterization that cannot be achieved with spinal cord homogenates. Such preparations of pure population of motor neurons will provide valuable tools to advance our understanding of the molecular mechanisms underlying spinal cord injury and neuromuscular diseases. In the near future, LCM techniques might be instrumental to the success of gene therapy for these debilitating conditions.


Ginkgolide B attenuates cerebral ischemia-reperfusion injury via inhibition of ferroptosis through disrupting NCOA4-FTH1 interaction.
Cerebral ischemia/reperfusion (I/R) injury is a major cause of neuronal damage and death. Ginkgolide B (GB) has been shown to exhibit neuroprotective effects in various brain injury models. The aim of study was to investigate the potential role of GB in protecting against cerebral I/R injury and explore the underlying mechanisms. Adult male Sprague-Dawley rats were exposed to transient middle cerebral artery occlusion (tMCAO) followed by reperfusion in order to trigger cerebral I/R injury. The rats were treated with different doses of GB, vehicle control or positive drug. Neurological function, infarct volume, and levels of ferroptosis markers were evaluated. In vitro experiments were performed using OGD/R-induced PC12 cells to further investigate the effects of GB on ferroptosis and its mechanisms. In addition, molecular docking, and microscale thermophoresis (MST) assay were conducted to explore the combination of GB and NCOA4. Reduced infarct volume and enhanced neurological function were signs of dose-dependent protection from cerebral I/R injury by GB therapy. Additionally, GB treatment had an impact on the levels of oxidative stress and ferroptosis markers, including reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD), and Fe2+ in the cerebral environment during IR injury. Moreover, relevant ferroptosis key factors such as ACSL4, GPX4, FTH1, and NCOA4 can be regulated by GB. In OGD/R-induced PC12 cells, GB protected against ferroptosis by inhibiting autophagy and disrupting the interaction of NCOA4-FTH1. Our findings suggest that GB may protect against cerebral I/R injury by inhibiting ferroptosis through disrupting NCOA4-FTH1 interaction. GB has potential therapeutic applications for cerebral I/R injury, and further investigation of the underlying mechanisms and clinical trials are warranted.


Modified technique using flexible iris retractors in clear corneal cataract surgery.
Flexible nylon iris retractors are a useful adjunct to cataract surgery in cases of small pupil. We present a modification of the standard technique. Instead of approaching the square-shaped pupil formed by iris retractors from the side, we recommend an approach anterior to 1 of the retractor hooks. From the surgeon's viewpoint, the pupil goes from a square shape to a diamond shape. This modification minimizes the likelihood of iris prolapse and iris damage during phacoemulsification and increases the workspace for the phacoemulsification needle.


Bronchial responsiveness to inhaled histamine in both adults with intrinsic and extrinsic asthma: the importance of prechallenge forced expiratory volume in 1 second.
Recent evidence suggests that both nonspecific bronchial hyperresponsiveness and degree of airflow obstruction may be involved in the deterioration of lung function observed in patients with bronchial asthma. To investigate to which degree the level of bronchial hyperresponsiveness reflects the size of the airways at the time of testing, we examined the relationship of nonspecific bronchial responsiveness to prechallenge pulmonary function in adult asthmatic patients. Bronchial responsiveness to inhaled histamine was measured in 100 patients (age range, 28 to 79 years), of whom 62 had intrinsic and 38 had extrinsic asthma. Histamine responsiveness was analyzed by means of the dose-response slope (DRS). No significant difference in prechallenge forced expiratory volume expressed as a percentage of predicted value (FEV1%pred), duration of asthma, smoking habits (pack years), or bronchial responsiveness (DRS) was found between the patients with intrinsic and those with extrinsic asthma. The patients had, in general, reduced pulmonary function, because 80% had a prechallenge FEV1 less than 80%pred. The DRS displayed a significant inverse relationship to the prechallenge level of FEV1%pred in both patients with intrinsic asthma and those with extrinsic asthma (p < 0.000001); the initial FEV1%pred accounted for approximately 35% of the variance in measurements of histamine responsiveness. Excluding patients with abnormal prechallenge FEV1 (< 70%pred), or current and exsmokers (never smoked: n = 67) from the analysis did not change these findings. Nonspecific bronchial responsiveness is to some extent determined by the prechallenge level of pulmonary function in both adults with intrinsic and extrinsic asthma.


Inhibition of prostacyclin formation by cyclosporin is not due to reduced availability of arachidonic acid in membrane phospholipids of cultured human endothelial cells.
Our studies have shown that CS inhibits PGI2 production in HUVEC, that this inhibition is not overcome when exogenous AA is supplied, that the inhibitory action of CS is proximal to PGI2 synthetase and finally that there is abundant free AA available in membrane phospholipids of CS treated HUVEC [4,5]. In conclusion, CS does not appear to inhibit PGI2 synthesis by reducing the availability of free AA in the endothelial cell membrane. Although CS appears to inhibit cyclo-oxygenase, we can not exclude an additional effect on acyltransferase.


Inference of age-dependent case-fatality ratios for seasonal influenza virus subtypes A(H3N2) and A(H1N1)pdm09 and B lineages using data from the Netherlands.
Despite the known relatively high disease burden of influenza, data are lacking regarding a critical epidemiological indicator, the case-fatality ratio. Our objective was to infer age-group and influenza (sub)type specific values by combining modelled estimates of symptomatic incidence and influenza-attributable mortality. The setting was the Netherlands, 2011/2012 through 2019/2020 seasons. Sentinel surveillance data from general practitioners and laboratory testing were synthesised to supply age-group specific estimates of incidence of symptomatic infection, and ecological additive modelling was used to estimate influenza-attributable deaths. These were combined in an Bayesian inferential framework to estimate case-fatality ratios for influenza A(H3N2), A(H1N1)pdm09 and influenza B, per 5-year age-group. Case-fatality estimates were highest for influenza A(H3N2) followed by influenza B and then A(H1N1)pdm09 and were highest for the 85+ years age-group, at 4.76% (95% credible interval [CrI]: 4.52-5.01%) for A(H3N2), followed by influenza B at 4.08% (95% CrI: 3.77-4.39%) and A(H1N1)pdm09 at 2.51% (95% CrI: 2.09-2.94%). For 55-59 through 85+ years, the case-fatality risk was estimated to double with every 3.7 years of age. These estimated case-fatality ratios, per influenza sub(type) and per age-group, constitute valuable information for public health decision-making, for assessing the retrospective and prospective value of preventative interventions such as vaccination and for health economic evaluations.


Enhanced air-cavity effect of periodically oriented embedded air protrusions for high-efficiency InGaN/GaN light-emitting diodes.
We report on the development of periodically oriented embedded air protrusion (EAP) structures at the GaN-sapphire interface in InGaN/GaN LEDs. A specific SiO(2) mask pattern and a simple wet etching process were utilized for the fabrication of EAP structures. A strong coupling between closely proximate air cavities and the multiple quantum wells promoted spontaneous emission due to the high-index contrast at the GaN-air interface. As a result, the light output power of the EAP LED was 2.2 times higher than that of a conventional LED at an injection current of 20 mA.


Synthesis of Optically Active α-Trifluoromethylamines by Rearrangement of β-Amino-α-trifluoromethyl Alcohols.
The synthesis of various optically active α-trifluoromethylamines has been realized from β-amino-α-trifluoromethyl alcohols via an aziridinium ion intermediate under kinetic conditions.


Hybrid UV/COP advanced oxidation process using ZnO as a catalyst immobilized on a stone surface for degradation of acid red 18 dye.
Azo dyes are the largest group of synthetic organic dyes which containing the linkage C-N[bond, double bond]N-C and used in various industries such as textile industries leather articles, and some foods. Azo dyes are resistant compounds against the biodegradation processes. The purpose of this research was hybrid UV/COP advanced oxidation process using ZnO as a catalyst immobilized on a stone surface for degradation of acid red 18 (AR18) Dye. In the hybrid process using some parameters such as the dye initial concentration, pH, contact time and catalyst concentration, the process efficiency was investigated. In order to the dye removal, the sole ozonation process (SOP), catalytic ozonation process (COP) and photocatalytic process (UV/ZnO) were used. The ZnO nanoparticles were characterized by XRD, SEM and TEM analyses. The maximum dye removal was achieved 97% at the dye initial concentration 25 mg/L, catalyst concentration 3 g/L, contact time 40 min and pH 5. As a real sample, the Yazdbaf textile factory wastewater was selected. After that, the physicochemical quality was evaluated. As well as, in the optimal conditions, the AR18 dye removal efficiency was achieved 65%. The kinetic results demonstrated that the degradation reaction was fitted by pseudo-first-order kinetic. The UV/COP hybrid process had high efficiency for removal of resistant dyes from the textile wastewater. Advantages of this technique were as follows:•ZnO nanoparticles were synthesized as catalyst by thermal method and were immobilized on the stones.•pH changes had no significant effect on the removal efficiency.•In the kinetic studies, the decomposition reaction followed pseudo-first order kinetic.


Forecasting the carbon dioxide emissions in 53 countries and regions using a non-equigap grey model.
Non-equigap GM(1,1) model with conformable fractional accumulation (CFNGM(1,1)) is proposed to analyze the relationship between energy consumption and carbon dioxide emissions. Two cases are used to prove the validity of the model. In this article, energy consumption is used as input and carbon dioxide emissions are used as output. Carbon dioxide emissions of 53 countries and regions in North America, South America, Europe, Commonwealth of Independent States (CIS), Middle East, Africa, and Asia Pacific are predicted. The forecast results show that the carbon dioxide emissions of 30 countries and regions have risen to varying degrees. The top three countries with carbon dioxide emissions in the next three years are China, the USA, and India. More attention should be paid to the carbon dioxide emissions of China.


Assessment of the incremental prognostic value from the modified frailty index-5 in complete traumatic cervical spinal cord injury.
Frailty, as measured by the modified frailty index-5 (mFI-5), and older age are associated with increased mortality in the setting of spinal cord injury (SCI). However, there is limited evidence demonstrating an incremental prognostic value derived from patient mFI-5. We conducted a retrospective cohort study to evaluate in-hospital mortality among adult complete cervical SCI patients at participating centers of the Trauma Quality Improvement Program from 2010 to 2018. Logistic regression was used to model in-hospital mortality, and the area under the receiver operating characteristic curve (AUROC) of regression models with age, mFI-5, or age with mFI-5 was used to compare the prognostic value of each model. 4733 patients were eligible. We found that both age (80 y versus 60 y: OR 3.59 95% CI [2.82 4.56], P < 0.001) and mFI-5 (score ≥ 2 versus < 2: OR 1.53 95% CI [1.19 1.97], P < 0.001) had statistically significant associations with in-hospital mortality. There was no significant difference in the AUROC of a model including age and mFI-5 when compared to a model including age without mFI-5 (95% CI Δ AUROC [- 8.72 × 10-4 0.82], P = 0.199). Both models were superior to a model including mFI-5 without age (95% CI Δ AUROC [0.06 0.09], P < 0.001). Our findings suggest that mFI-5 provides minimal incremental prognostic value over age with respect to in-hospital mortality for patients complete cervical SCI.


Foot and ankle Osteoarthritis and Cognitive impairment in retired UK Soccer players (FOCUS): protocol for a cross-sectional comparative study with general population controls.
Professional footballers commonly experience sports-related injury and repetitive microtrauma to the foot and ankle, placing them at risk of subsequent chronic pain and osteoarthritis (OA) of the foot and ankle. Similarly, repeated heading of the ball, head/neck injuries and concussion have been implicated in later development of neurodegenerative diseases such as dementia. A recent retrospective study found that death from neurodegenerative diseases was higher among former professional soccer players compared with age matched controls. However, well-designed lifetime studies are still needed to provide evidence regarding the prevalence of these conditions and their associated risk factors in retired professional football players compared with the general male population. To determine whether former professional male footballers have a higher prevalence than the general male population of: (1) foot/ankle pain and radiographic OA; and (2) cognitive and motor impairments associated with dementia and Parkinson's disease. Secondary objectives are to identify specific football-related risk factors such as head impact/concussion for neurodegenerative conditions and foot/ankle injuries for chronic foot/ankle pain and OA. This is a cross-sectional, comparative study involving a questionnaire survey with subsamples of responders being assessed for cognitive function by telephone assessment, and foot/ankle OA by radiographic examination. A sample of 900 adult, male, ex professional footballers will be recruited and compared with a control group of 1100 age-matched general population men between 40 and 100 years old. Prevalence will be estimated per group. Poisson regression will be performed to determine prevalence ratio between the populations and logistic regression will be used to examine risk factors associated with each condition in footballers. This study was approved by the East Midlands-Leicester Central Research Ethics Committee on 23 January 2020 (REC ref: 19/EM/0354). The study results will be disseminated at national and international meetings and submitted for peer-review publication.


Effect of cellular aging on collagen biosynthesis: I. Methodological considerations and pharmacological applications.
The study of the age and passage dependent modifications of collagen biosynthesis requires a simple, rapid and reproducible procedure adaptable to serial cell cultures. To make such a method comparable to other methods of collagen determination, we calibrated a colorimetric procedure both by hydroxyproline (HYP) determinations and in terms of collagen concentration. For collagen types I and IV, widely different slopes were obtained with the colorimetric procedure. To further refine the procedure, we tempted to completely inhibit collagen synthesis by beta-aminopropionitrile (beta APN) added to cultures in order to obtain a negative control. Using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl 2H-tetrazolium bromide (MTT-test), it could be shown that relatively high concentrations of beta APN are tolerated by the cells. It appeared, however, that even the highest concentration of beta APN (1mM) still tolerated by the fibroblasts did not completely inhibit collagen synthesis. At low concentrations, beta APN even stimulated cell-proliferation. The colorimetric procedure calibrated in terms of collagen type I concentration, was therefore retained for the serial determination of collagen synthesis and accumulation. We shall here describe the methodological details of its validation as well as its application for the pharmacological study of the effect of aging on collagen biosynthesis. Among the factors involved, the accumulation of advanced glycation end-products (AGEs) might well play an important role. Several of such AGE-products showed a significant inhibition of collagen deposition. On the contrary, retinol, ascorbic acid as well as the rhamnose-rich oligo- and polysaccharides (RROPs) did produce a significant upregulation collagen deposition. Polysaccharide preparations, rich in rhamnose and fucose (the EROB-mixture) could protect against the AGEs-induced inhibition of collagen accumulation.


Degradation of levofloxacin from antibiotic wastewater by pulse electrochemical oxidation with BDD electrode.
Antibiotic-containing wastewater is a typical biochemical refractory organic wastewater and general treatment methods cannot effectively and quickly degrade the antibiotic molecules. In this study, a novel boron-doped diamond (BDD) pulse electrochemical oxidation (PEO) technology was proposed for the efficient removal of levofloxacin (LFXN) from wastewater. The effects of current density (j), initial pH (pH0), frequency (f), electrolyte types and initial concentration (c0(LFXN)) on the degradation of LFXN were systematically investigated. The degradation kinetics under four different processes have also been studied. The possible degradation mechanism of LFXN was proposed by Density functional theory calculation and analysis of degradation intermediates. The results showed that under the optimal parameters, the COD removal efficiency (η(COD)) was 94.4% and the energy consumption (EEC) was 81.43 kWh·m-3 at t = 120 min. The degradation of LFXN at pH = 2.8/c(H2O2) followed pseudo-first-order kinetics. The apparent rate constant was 1.33 × 10-2 min-1, which was much higher than other processes. The degradation rate of LFXN was as follows: pH = 2.8/c(H2O2) > pH = 2.8 > pH = 7/c(H2O2) > pH = 7. Ten aromatic intermediates were formed during the degradation of LFXN, which were further degraded to F-, NH4+, NO3-, CO2 and H2O. This study provides a promising approach for efficiently treating LFXN antibiotic wastewater by pulsed electrochemical oxidation with a BDD electrode without adding H2O2.


Thrombin induces DNA synthesis and phosphoinositide hydrolysis in airway smooth muscle by activation of distinct receptors.
Chronic airway inflammation induces numerous structural changes of the airways involving hypertrophy and hyperplasia of airway smooth muscle (ASM). Thrombin has been identified in the bronchoalveolar lavage fluid of asthmatic subjects and displays potent bronchoconstrictor and mitogenic activity towards ASM. This study has addressed which proteinase-activated receptors (PARs) and signalling pathways are involved in mediating distinct effects of thrombin. Using cultured bovine tracheal smooth muscle (BTSM) cells as a model system, thrombin stimulated a marked increase in [3H]inositol phosphate ([3H]InsPs) accumulation, which was fully mimicked by a selective PAR1 activating peptide. In contrast, PAR1, PAR2, PAR3 and PAR4 activating peptides were unable to replicate the ability of thrombin to stimulate DNA synthesis as assessed by [3H]thymidine incorporation. Further investigation demonstrated that the mitogenic effect of thrombin did not involve stimulation of PDGF secretion but did involve activation of PDGF or EGF receptors and a G(i/o)-dependent activation of phosphoinositide 3-kinase. Thrombin, but not the PAR1, PAR2, PAR3 or PAR4 activating peptides was able to stimulate PtdIns(3,4,5)P3 mass accumulation. PAR3 antisense oligonucleotides substantially inhibit thrombin-stimulated [3H]thymidine incorporation and PtdIns(3,4,5)P3 generation but had no effect on thrombin-induced phosphoinositide hydrolysis. These data indicate that while PI hydrolysis and Ca2+ mobilisation induced by thrombin operates via PAR1-dependent activation of phospholipase C, phosphoinositide 3-kinase activation and DNA synthesis occurs via a distinct proteinase-activated receptor pathway, possibly involving PAR3.


Applications of Artificial Intelligence in Drug Design: Opportunities and Challenges.
Artificial intelligence (AI) has undergone rapid development in recent years and has been successfully applied to real-world problems such as drug design. In this chapter, we review recent applications of AI to problems in drug design including virtual screening, computer-aided synthesis planning, and de novo molecule generation, with a focus on the limitations of the application of AI therein and opportunities for improvement. Furthermore, we discuss the broader challenges imposed by AI in translating theoretical practice to real-world drug design; including quantifying prediction uncertainty and explaining model behavior.


Influence of cleavage-stage embryo quality on the in-vitro fertilization outcome after single embryo transfer in fresh cycles.
Embryo quality is crucial for determining the outcome of embryo implantation. This study aimed to assess the impact of embryo quality on the outcome of in vitro fertilization/single-embryo transfer (IVF-SET). This retrospective study included 2531 fresh IVF-SET cycles, including 277 poor-quality and 2254 top-quality embryos. The clinical pregnancy rate, miscarriage rate, live birth, implantation rate, pregnancy outcome and complication were analyzed and compared. Risk factors associated with miscarriage rate and pregnancy complication were identified using logistics regression analysis. Top-quality embryos resulted in higher clinical pregnancy rate (30.5% vs. 12.6%, P < 0.001) and live birth rate (23.9% vs. 9.7%, P < 0.001) compared with poor-quality embryos. Logistics regression analysis revealed that embryo quality was not correlated with miscarriage rate (95% CI 0.33-1.89) and pregnancy complications (95% CI 0.12-7.84). Maternal age and body mass index was a risk factor for miscarriage rate (95% CI 1.05-1.22) and pregnancy complication (95% CI 1.01-1.29), respectively. Clinical miscarriage rate and pregnancy complication were embryo quality independent. Maternal age was the risk factor for miscarriage rate. Embryo quality did not affect miscarriage once a clinical pregnancy is achieved.


Surgical treatment of biliary atresia in the liver transplantation era.
Biliary atresia (BA) still remains one of the most intractable gastrointestinal diseases in infancy despite the concerted efforts of pediatric surgeons all over the world. The introduction of liver transplantation has revolutionized the protocols for the treatment of this condition. In this editorial, the role of hepatic portoenterostomy (the Kasai procedure) in the surgical treatment of BA in the "transplantation era" will be discussed.


Geriatric pharmacotherapy: Appraising new drugs for neurologic disorders in older patients.
New drug development for neurologic disorders has one of the highest attrition rates of all clinical drug developments. This is problematic when, with innovative technology available in so many aspects of life, modern societies expect to have effective treatments for neurodegenerative disorders and mental health conditions that provide something beyond simple symptomatic relief-the expectation is treatment that impacts and mitigates fundamental mechanisms that drive these disorders. The disease burden of neurologic disorders remains extremely high, whereas the proportion of patients receiving effective therapy is relatively low, demonstrating a sizeable unmet medical need. Whether for novel breakthrough therapies or for drugs considered successful, deciding on the basis of clinical trial data whether a particular treatment will be effective for a specific patient is always a leap of faith. However, expertise at reading trial results combined with knowledge of the patient and of his or her disease, together with an understanding of the effect of age on drug pharmacokinetics and pharmacodynamics, the effect of age on the patient's condition, and the effect of age on the patient's life and outlook will ensure the landing is safe. The focus of this article is to provide such knowledge and thereby optimize this expertise.


Evaluation of two in vitro assays for tumorigenicity assessment of CRISPR-Cas9 genome-edited cells.
Off-target editing is one of the main safety concerns for the use of CRISPR-Cas9 genome editing in gene therapy. These unwanted modifications could lead to malignant transformation, which renders tumorigenicity assessment of gene therapy products indispensable. In this study, we established two in vitro transformation assays, the soft agar colony-forming assay (SACF) and the growth in low attachment assay (GILA) as alternative methods for tumorigenicity evaluation of genome-edited cells. Using a CRISPR-Cas9-based approach to transform immortalized MCF10A cells, we identified PTPN12, a known tumor suppressor, as a valid positive control in GILA and SACF. Next, we measured the limit of detection for both assays and proved that SACF is more sensitive than GILA (0.8% versus 3.1% transformed cells). We further validated SACF and GILA by identifying a set of positive and negative controls and by testing the suitability of another cell line (THLE-2). Moreover, in contrast to SACF and GILA, an in vivo tumorigenicity study failed to detect the known tumorigenic potential of PTPN12 deletion, demonstrating the relevance of GILA and SACF in tumorigenicity testing. In conclusion, SACF and GILA are both attractive and valuable additions to preclinical safety assessment of gene therapy products.


Factors associated with the effect of pulmonary rehabilitation on physical activity in patients with chronic obstructive pulmonary disease.
Although the effects of pulmonary rehabilitation (PR) have been well defined for chronic obstructive pulmonary disease (COPD), it remains controversial whether PR improves physical activity (PA). The purpose of the present study was to identify factors associated with the effect of PR on PA. This was a prospective study of 29 patients with COPD. They underwent pulmonary rehabilitation twice weekly for 12 weeks, and were assessed using the hospital anxiety and depression score, 6-min walk distance (6MWD), and the St. George Respiratory Questionnaire (SGRQ) before and after they underwent PR. The PA of patients was measured by a three-axis accelerometer. Physical activity level (PAL) was calculated by dividing each patient's total energy expenditure by basal metabolic rate. Correlations between changes in PAL after PR and 6MWD, St. George Respiratory Questionnaire, and hospital anxiety and depression score scores, and clinical parameters, including forced expiratory volume in 1 s were determined. 6MWD was significantly increased, but PAL was unchanged after PR. PAL was positively correlated with 6MWD, but not with percent predicted forced expiratory volume in 1 s nor St. George Respiratory Questionnaire scores before PR. The increase in PAL was negatively correlated with changes in hospital anxiety and depression score anxiety and depression scores, but was not correlated with the change in 6MWD. A PR program for COPD patients improved results of the 6MWD, but not PAL. Increased PAL was associated with improvements in anxiety and depression, but not with increased exercise capacity. Treating the depression and anxiety of patients with COPD might not only reduce emotional distress, but also improve their PAL. Geriatr Gerontol Int 2017; 17: 17-23.


Disrupted functional network in patients with temporal lobe epilepsy with impaired alertness.
Cognitive impairment is common in patients with temporal lobe epilepsy (TLE). Alertness is an important subfunction of cognition, but it is poorly understood in TLE. We hypothesized that disruptions to underlying brain networks may affect alertness in patients with TLE. Patients with unilateral TLE were grouped into low-alertness and high-alertness groups, and they were matched with healthy controls (HCs) (n = 20 each). Functional magnetic resonance imaging (fMRI) was used to construct functional brain networks, and graph theory was used to identify topological parameters of the networks. At the global level, patients with low alertness had networks with less small-worldness and less normalized clustering than HCs. At the nodal level, patients with low alertness exhibited decreased centrality of the bilateral parahippocampal gyrus compared with HCs and increased centrality of the right postcentral gyrus compared with patients with high alertness. This study reveals a decreased separation, tending toward randomization, of the functional network in patients with TLE with impaired alertness. Our results also suggest that the parahippocampal gyrus may contribute to impaired alertness and the right postcentral gyrus plays an important role in the modulation of alertness in patients with TLE.


Gypenosides derived from Gynostemma pentaphyllum suppress NO synthesis in murine macrophages by inhibiting iNOS enzymatic activity and attenuating NF-kappaB-mediated iNOS protein expression.
Gypenosides isolated from Gynostemma pentaphyllum are widely used in traditional Chinese medicine, with beneficial effects reported in numerous diseases, including inflammation and atherosclerosis, although the mechanism underlying these therapeutic effects is unknown. Because increased nitric oxide (NO) plays a role in these pathological conditions, we investigated whether the pharmacological activity of gypenosides is due to suppression of NO synthesis. The markedly increased production of nitrite by stimulation of RAW 264.7 murine macrophages with 1 microg/mL lipopolysaccharide (LPS) for 20 h (unstimulated: 0.3+/-0.3 microM vs. LPS: 32.5+/-1.2 microM) was dose-dependently inhibited by gypenosides (0.1-100 microg/mL). When cells were pretreated with gypenosides (for 1h) prior to LPS stimulation, subsequent NO production was significantly attenuated (IC(50) of 3.1+/-0.4 microg/mL) (P<0.05). Gypenosides (25 microg/mL) produced the same maximum inhibition of LPS-induced NO production as aminoguanidine, a standard inhibitor of NOS enzymes. Suppression of NO production occurred both by direct inhibition of the activity and expression of iNOS. Inhibition of iNOS protein expression appears to be at the transcriptional level, since gypenosides decreased LPS-induced NF-kappaB activity in a dose-dependent manner (P<0.05), with significant inhibition achieved following pretreatment with 10 microg/mL gypenoside. Taken together, these results suggest that gypenosides derived from G. pentaphyllum suppress NO synthesis in murine macrophages by inhibiting iNOS enzymatic activity and attenuating NF-kappaB-mediated iNOS protein expression, thereby implicating a mechanism by which gypenosides may exert their therapeutic effects.


S4153R is a gain-of-function mutation in the cardiac Ca(2+) release channel ryanodine receptor associated with catecholaminergic polymorphic ventricular tachycardia and paroxysmal atrial fibrillation.
Mutations in ryanodine receptor 2 (RYR2) gene can cause catecholaminergic polymorphic ventricular tachycardia (CPVT). The novel RYR2-S4153R mutation has been implicated as a cause of CPVT and atrial fibrillation. The mutation has been functionally characterized via store-overload-induced Ca(2+) release (SOICR) and tritium-labelled ryanodine ([(3)H]ryanodine) binding assays. The S4153R mutation enhanced propensity for spontaneous Ca(2+) release and reduced SOICR threshold but did not alter Ca(2+) activation of [(3)H]ryanodine binding, a common feature of other CPVT gain-of-function RYR2 mutations. We conclude that the S4153R mutation is a gain-of-function RYR2 mutation associated with a clinical phenotype characterized by both CPVT and atrial fibrillation.


Apparent prevalence of antibodies to Coxiella burnetii (Q fever) in bulk tank milk from dairy herds in southern Belgium.
In 2006, a cross-sectional survey was carried out in 206 herds in southern Belgium to estimate the proportion of herds with Q fever-seropositive bulk tank milk (BTM), i.e. herds with an intra-herd seroprevalence >10%. Of the tested herds, 119 (57.8%; 95% CI: 51.1-64.4%) were seropositive. Most had a low antibody titre. Of these herds, 50 were randomly subjected to a real time Coxiella burnetii PCR analysis. The proportion of herds excreting Coxiella in BTM was 30.0% with only 2.0% of herds having a high level of shedding. An additional questionnaire and logistical regression identified some factors which increased the risk of BTM seroconversion (in heifers, drinking water from watercourse or well water) and some which decreased it (in heifers, tie- and free-stalling, and animals of all age in the same shed; in heifers and cows, shed disinfection).


Unveiling the mechanisms of medium-chain fatty acid production from waste activated sludge alkaline fermentation liquor through physiological, thermodynamic and metagenomic investigations.
Effective sludge treatment with bioenergy production is attracting increasing interests as large quantities of waste activated sludge (WAS) are produced during the wastewater treatment. In this study, a new biotechnical process for converting the WAS alkaline fermentation liquor (WASAFL) into valuable, easy-separated medium chain fatty acids (MCFAs) through chain elongation (CE) was investigated, which may provide a new insight into sludge treatment. In the process, ethanol was served as the electron donor (EDs) and WASAFL were main electron acceptors (EAs). The MCFAs productions were investigated under three different ED to EA ratios (i.e., 1:2, 1:1 and 2:1). The result showed that MCFAs production was increased from 2.88 ± 0.01 to 5.28 ± 0.18 g COD/L with the increase of ED to EA ratio. However, the highest MCFA selectivity was achieved at 72.9% when the ED to EA ratio was 1:1. The decrease in the selectivity at high ED:EA ratio is mainly due to the production of higher alcohol (i.e., n-butanol and n-hexanol). The thermodynamic analysis confirmed all CE processes for MCFAs production from WASAFL were exothermic reactions, with the spontaneity and energy release of the reactions increased with the ethanol level. The microbial community analysis showed that the relative abundances of Clostridium, Oscillibacter, Leptolinea and Exilispira were positively correlated with the MCFAs production. The metagenomic analysis suggested that both the reverse β-oxidization pathway and fatty acid biosynthesis pathway contributed to the CE process in the studied system. The functional enzymes were mainly associated within Clostridium, with Clostridium Kluyveri, Clostridium botulinum and Clostridium magnum being likely the key species responsible for the CE process.


Nutritional physiology and selective isolation of Exophiala dermatitidis.
The nutritional physiology of ten strains of Exophiala dermatitidis was investigated. The growth reactions to lactose, citrate, nitrate, nitrite, lysine, creatine and creatinine differ from those found in closely related black yeasts. In addition, it is the only Exophiala species which is able to grow at 40 degrees C. A selective medium containing meso-erythritol as sole carbon source was evaluated. This medium is particularly useful for the isolation of black yeasts from lungs of patients with cystic fibrosis as well as from the environment.