Localization and characterization of gonadotropin-releasing hormones in the brain, gonads, and plasma of a dipnoi (lungfish, Protopterus annectens).
Two molecular forms of GnRH (chicken GnRH II and a second variant) are present in the brains of species from all the major vertebrate groups. Their differential distribution in the brain and temporal expression during development suggests that have different functional roles. We investigated the nature of GnRH molecular forms in the brain, plasma, testis, and ovary of adult and juvenile lungfish (Protopterus annectens), using high performance liquid chromatography and radioimmunoassay with specific GnRH antisera. In the brain of adult and juvenile lungfish, two peptides with identical chromatographic and immunologic properties to mammalian GnRH and chicken GnRH II were detected. Chicken GnRH II predominated in both the adult and juvenile brain, and the percentage of chicken GnRH II relative to mammalian GnRH was greater in the juvenile brain. In the plasma, only mammalian GnRH was present. Immunoreactive GnRH was not detected in the testis and ovary. Chicken GnRH II and mammalian GnRH were found in the cells of the preoptic nucleus and in the ganglion of the nervus terminalis. Fibers were seen in the ventral hypothalamus, and chicken GnRH II immunoreactivity was detected within the neural lobe of the pituitary. The finding of chicken GnRH II in a sarcopterygian fish adds further support to our hypothesis that this ubiquitous structural variant is highly conserved and likely to have an important functional role. Mammalian GnRH, previously described in several early-evolved actinopterygian fish, also has a fairly widespread distribution and early evolutionary origin. The immunocytochemical distribution of mammalian GnRH and chicken GnRH II fibers in the lungfish brain suggests that both forms are hypophysiotropic. In addition, the presence of mammalian GnRH in the plasma of the lungfish suggests that this molecular form of GnRH has a hypophysiotropic function reaching target organs (pituitary and gonads) via the general circulation.


Computer-based classification of dermoscopy images of melanocytic lesions on acral volar skin.
We describe a fully automated system for the classification of acral volar melanomas. We used a total of 213 acral dermoscopy images (176 nevi and 37 melanomas). Our automatic tumor area extraction algorithm successfully extracted the tumor in 199 cases (169 nevi and 30 melanomas), and we developed a diagnostic classifier using these images. Our linear classifier achieved a sensitivity (SE) of 100%, a specificity (SP) of 95.9%, and an area under the receiver operating characteristic curve (AUC) of 0.993 using a leave-one-out cross-validation strategy (81.1% SE, 92.1% SP; considering 14 unsuccessful extraction cases as false classification). In addition, we developed three pattern detectors for typical dermoscopic structures such as parallel ridge, parallel furrow, and fibrillar patterns. These also achieved good detection accuracy as indicated by their AUC values: 0.985, 0.931, and 0.890, respectively. The features used in the melanoma-nevus classifier and the parallel ridge detector have significant overlap.


Author Correction: In situ 3D-patterning of electrospun fibers using two-layer composite materials.
An amendment to this paper has been published and can be accessed via a link at the top of the paper.


Three-dimensional print of a liver for preoperative planning in living donor liver transplantation.
The growing demand for liver transplantation and the concomitant scarcity of cadaveric livers have increased the need for living donor liver transplantation (LDLT). Ensuring the safety of donors and recipients is critical. The preoperative identification of the vascular and biliary tract anatomy with 3-dimensional (3D) printing may allow better preoperative surgical planning, avert unnecessary surgery in patients with potentially unsuitable anatomy, and thereby decrease the complications of liver transplant surgery. We developed a protocol and successfully 3D-printed synthetic livers (along with their complex networks of vascular and biliary structures) replicating the native livers of 6 patients: 3 living donors and 3 respective recipients who underwent LDLT. To our knowledge, these are the first complete 3D-printed livers. Using standardized preoperative, intraoperative, and postoperative assessments, we demonstrated identical anatomical and geometrical landmarks in the 3D-printed models and native livers.


Dermal absorption and penetration of jet fuel components in humans.
Jet propulsion fuel 8 (JP-8) is the largest source of chemical exposures on military bases. Dermal exposure to JP-8 has been investigated in vitro using rat or pig skin, but not in vivo in humans. The purpose of this study was to investigate the absorption and penetration of aromatic and aliphatic components of JP-8 in humans. A surface area of 20 cm2 was delineated on the forearms of human volunteers and 1 ml of JP-8 was applied to the skin. Tape-strip samples were collected 30 min after application. Blood samples were taken before exposure (t=0 h), after exposure (t=0.5 h), and every 0.5 h for up to 4 h past exposure. The tape-strip samples showed evidence of uptake into the skin for all JP-8 components. The blood data was used to estimate an apparent permeability coefficient (Kp). The rank order of the apparent Kp was naphthalene>1-methyl naphthalene=2-methyl naphthalene>decane>dodecane>undecane. This rank order is similar to results from rat and pig-skin studies. However, this study demonstrates that rat and pig models of the skin over predict the internal dose of JP-8 components in humans.


Does metformin modify the effect on glycaemic control of aerobic exercise, resistance exercise or both?
Some previous studies suggested that metformin might attenuate the effects of exercise on glycaemia or fitness. We therefore examined whether metformin use influenced changes in glycaemic control, fitness, body weight or waist circumference resulting from aerobic and/or resistance training in people with type 2 diabetes participating in an exercise intervention trial. After a 4 week run-in period, participants from the Diabetes Aerobic and Resistance Exercise (DARE) trial were randomly assigned to 22 weeks of aerobic training alone, resistance training alone, combined aerobic and resistance exercise training or a waiting-list control group. Of the 251 randomised, 143 participants reported using metformin throughout the entire study period and 82 reported not using metformin at all. Compared with control, aerobic training led to a significant reduction in HbA1c in the metformin users (-0.57%, 95% CI -1.05, -0.10; -6.3 mmol/mol, 95% CI -11.5, -1.1) but not in the non-metformin users (-0.17, 95% CI -0.78, 0.43; -1.9 mmol/mol, 95% CI -8.5, 4.7). However, there were no significant differences in the changes in HbA1c (or fasting glucose) between metformin users and non-users in any of the exercise groups compared with control (p> 0.32 for all metformin by group by time interactions). Similarly, metformin did not affect changes in indicators of aerobic fitness, strength and body weight or waist circumference (p ≥ 0.15 for all metformin by group by time interactions). Contrary to our hypothesis and to previous short-term studies, metformin did not significantly attenuate the benefits of exercise on glycaemic control or fitness.


Elevated glucose alters paracellular transport of cultured human proximal tubule cells.
Cultures of human proximal tubule cells were exposed to elevated concentrations of glucose and dome formation was assessed over a 22 day period of growth. Cultures grown on 5.5 mM glucose formed five domes per microscopic field while those exposed to elevated glucose concentrations (11.0 mM to 27.5 mM) formed only two to three domes per field. The areas of the domes formed by the cells grown on elevated glucose concentrations were reduced as compared to those formed on 5.5 mM glucose. An analysis of the electrical properties of cells grown on elevated glucose concentrations by Ussing chamber technique disclosed a marked reduction in potential difference, short circuit current, and resistance compared to cells grown on 5.5 mM glucose. Routine ultrastructural analysis disclosed that cells grown on elevated glucose concentrations appeared to have fewer tight junction complexes. Further examination utilizing freeze fracture methodology demonstrated that cells grown on elevated glucose concentrations averaged two to three sealing strands per junction as compared to an average of five sealing strands for cells grown on 5.5 mM glucose. The cells grown on elevated glucose concentrations were also noted to possess a greater number of gap junctions. These results demonstrate that elevated glucose concentrations can alter the paracellular route, and possibly the transcellular route, of transport regulation in cultured human proximal tubule cells.


Evidence of determinant spreading in the antibody responses to prostate cell surface antigens in patients immunized with prostate-specific antigen.
Prostate cancer consistently remains a difficult clinical problem. The development of novel therapy strategies for effective control and treatment of prostate cancer is essential. The prostate represents a unique site for immunotherapy, in part because prostate-specific immunity would most probably be without significant long-term sequellae. Antibodies and cell-mediated immunity, induced by either active or passive immunization, represent potential means to specifically target prostate tumor cells. The serum IgG response to cell surface antigens expressed on LNCAP [prostate-specific antigen (PSA)-positive] and PC-3 (PSA-negative) were analyzed in individuals with advanced disease receiving vaccinia- or fowlpox-expressed PSA (v-PSA or f-PSA, respectively) by flow cytometry. Sera from all seven patients in a Phase I study of v-PSA, collected prior to the third immunization, reacted with both prostate tumor cell lines. The majority of individuals (n = 12) in a Phase II trial of v-PSA and f-PSA developed sustainable antibody responses to cell surface antigens on the prostate tumor cell lines. The magnitude and kinetics of these responses were dependent on the immunization schedule. Of importance, the baseline serum of only one of nine patients tested had reactivity with nonprostate tumor cell lines. Sera from three normal males also lacked reactivity with prostate tumor cells. PSA vaccine constructs are immunogenic and induce antibody responses to a multitude of surface antigens on prostate tumor cell lines by epitope or determinant spreading after stimulation of the immune system by PSA immunization.


Comparative studies on the effect of 4-APP(4-aminopyrazolopyrimidine) on hamster and rat adrenal cortex.
The study aimed to compare, by means of stereologic methods, the reactivity of the adrenal cortex of the hamster and the rat to prolonged treatment with 4-aminopyrazolo-(3,4d)pyrimidine (4-APP), a drug reducing hepatic secretion of plasma lipoprotein. Adult female hamsters, intact or cortisone suppressed, were administered i.p. daily with 0.5 mg 4-APP for 5 days while intact female rats received 1 mg of the drug per dose. 4-APP resulted in a loss of body weight, with the more profound effect in the hamster. In the rat 4-APP did not change the adrenal gland weight, the volume of the adrenocortical zones, the average volume of adrenocortical cells and the number of parenchymal cells in the gland. Moreover, serum ACTH and corticosterone levels in the rat remained unchanged. In the intact hamster 4-APP decreased the adrenal gland weight, the volume of the zona fasciculata and enhanced the serum cortisol level. In steroid suppressed hamsters 4-APP lowered the adrenal weight, the volume of fasciculata and reticularis zones, the average volume of the fasciculata cells and the number of parenchymal cells in the gland. These findings may suggest the direct inhibitory effect of low 4-APP doses on the hamster adrenal cortex and clearly demonstrate higher susceptibility of the hamster adrenal cortex, if compared with the rat, to 4-aminopyrazolo-pyrimidine.


Preferential localisation of red cell vacuoles to pathologically shaped human red blood cells.
The percentage of pitted erythrocytes and Howell-Jolly bodies in peripheral blood samples of 51 individuals following posttraumatic splenectomy and 20 patients splenectomized because of various haematological diseases differed significantly from each other (p less than 0.001) and from that of healthy controls (p less than 0.001). The percentage of pitted erythrocytes was significantly higher in pathologically shaped red blood cells (RBCs) (acanthocytes, schizocytes, elliptocytes) than in normal discoid shaped RBCs (p less than 0.001). As the number of pits per RBC showed great individual variations, a scoring system for the evaluation of pitted RBCs is proposed.


Biology and evolution of bacterial toxin-antitoxin systems.
Toxin-antitoxin systems are widespread in bacterial genomes. They are usually composed of two elements: a toxin that inhibits an essential cellular process and an antitoxin that counteracts its cognate toxin. In the past decade, a number of new toxin-antitoxin systems have been described, bringing new growth inhibition mechanisms to light as well as novel modes of antitoxicity. However, recent advances in the field profoundly questioned the role of these systems in bacterial physiology, stress response and antimicrobial persistence. This shifted the paradigm of the functions of toxin-antitoxin systems to roles related to interactions between hosts and their mobile genetic elements, such as viral defence or plasmid stability. In this Review, we summarize the recent progress in understanding the biology and evolution of these small genetic elements, and discuss how genomic conflicts could shape the diversification of toxin-antitoxin systems.


The relationships between oral language and reading instruction: Evidence from a computational model of reading.
Reading acquisition involves learning to associate visual symbols with spoken language. Multiple lines of evidence indicate that instruction on the relationship between spellings and sounds may be particularly important.However, it is unclear whether the effectiveness of this form of instruction depends on pre-existing oral language knowledge.To investigate this issue, we developed a series of computational models of reading incorporating orthographic, phonological and semantic processing to simulate bothartificialand natural orthographic learning conditions in adults and children. We exposed the models to instruction focused on spelling-sound or spelling-meaning relationships, and tested the influence of the models' oral language proficiency on the effectiveness of these training regimes. Overall, the simulations indicated thatoral language proficiency is a vital foundation for reading acquisition, and may modulate the effectiveness of reading instruction. These results provide a computational basis for the Simple View of Reading,and emphasise the importance of both oral language knowledge and spelling-sound instructionin the initial stages of learning to read.


The lateral wrist radiograph - To retake or not to retake.
Patient positioning may impact diagnostic quality when obtaining radiographs of the musculoskeletal (MSK) system. Hence, knowledge on patient positioning, as seen in the radiograph, followed by informed adjusted retake if appropriate, is key when undertaking MSK radiographs. Forearm positioning is particularly important in lateral wrist radiographs where rotation impacts anatomic measurements. The purpose was to evaluate the accuracy of MSK and non-MSK radiographers' immediate assessments of wrist positioning including need for retake. A questionnaire including images of 18 lateral wrist radiographs and questions regarding positioning, i.e. forearm rotation and flexion of the wrist, were developed and distributed to radiographers worldwide via the European Federation of Radiographer Societies (EFRS) and the Research Hub at the European Congress of Radiology (ECR) 2021. Demographic data such as area of expertise, years of experience etc. were collected. In total, 156 replies were included in the analyses. The inter-observer agreement of radiographers' assessment of the need for a retake was 47% (kappa = .25) and the intra-observer agreement was 81% (kappa = .62). Radiographers working with MSK radiography had more correct positioning assessments than radiographers who did not routinely obtain radiographs of the MSK system (p = 0.0003). Results indicated that MSK radiographers are more consistent in assessment of the need for a retake in lateral wrist radiographs and more able to correctly judge positioning compared to non-MSK radiographers. Constant focus on image quality may lead to increased awareness and adherence to image criteria. Improved image quality will in turn improve the diagnostic value for the benefit of the patients potentially leading to better outcomes.


Survival of patients with incident dementia who had a pre-existing psychiatric disorder: a population-based 7-year follow-up study.
Although it is widely accepted that psychiatric disorders and dementia coexist and survival data for dementia patients have been published, there is a paucity of information regarding the survival of patients with a psychiatric disorder who develop dementia. This study fills this information gap providing survival data on patients with such comorbidity and identifies mortality risk factors. All residents of Saskatchewan, a Canadian province, diagnosed with psychiatric problems and/or dispensed a psychiatric drug in 2000 and without dementia were followed through to 31 December 2006; the development of incident dementia was noted. Median survival time (in months) and selected predictors of mortality were measured. Analyses used Cox's proportional hazard model. Incidence density of dementia for the year 2000 was also computed. By December 2006, 5,583 subjects with psychiatric disorders in 2000 had been diagnosed with incident dementia, and 60.65% of them died. Dementia-incidence density in this population for 2000 was 0.01 per 1000 person years at risk among those aged 18-64 years and rapidly increased to 3.13 per 1000 person years at risk among those aged 75 to 84 years. The median survival time from dementia onset to death was 32.66 months (interquartile range 31.21-34.14). Being male, later age of onset of dementia, having a lower income, and a high chronic disease score predicted shorter survival. The comorbidity of psychiatric disorders and dementia resulted in shorter survival compared with that reported for patients with dementia only. These findings can be used for prognosis for patients, caregivers, and service providers.


Targeted inhibition of genome-wide DNA methylation analysis in epigenetically modulated phenotypes in lung cancer.
DNA methylation analysis, an epigenetic specification, has been explored for partial determination of cancer cell phenotypes. The development of metastasis in cancerogenesis has led its feasible association with the epigenetic modulations. We generated highly aggressive non-small cell lung cancer cell lines (HTB56 and A549) by using in vivo selection approach. These were, then, subjected to DNA methylation analysis (genome-wide). We also explored the therapeutic effects of azacytidine, an epigenetic agent, on DNA methylation patterns as well as the in vivo phenotypes. During the development of highly aggressive cell lines, we observed widespread modulations in DNA methylation. Reduced representation bisulfite sequencing was used and compared with the less aggressive parental cell lines to identify the differential methylation, which was achieved up to 2.7 % of CpG-rich region. Azacytidine inhibited DNA methyltransferase and reversed the prometastatic phenotype. We found its high association with the preferential loss of DNA methylation from hypermethylated sites. After persisted exposure of azacytidine, we observed that DNA methylation affected the polycomb-binding sites. We found close association of DNA methylome modifications with metastatic capability of non-small cell lung cancer. We also concluded that epigenetic modulation could be used as a potential therapeutic approach to prevent metastasis formation as prometastatic phenotype was reversed due to inhibition of DNA methyltransferase.


Comparison of astrocytes and gap junction proteins in the white matter of genetic absence epileptic and control rats: an experimental study.
The role of white matter astrocytes in absence epilepsy is unknown. The present study aims to quantify astrocytic markers glial fibrillary acidic protein (GFAP), gap junction's proteins connexin 30 (Cx30) and connexin 43 (Cx43) in the corpus callosum (CC) of genetic absence epileptic rats from Strasbourg (GAERS), Wistar albino glaxo rats from Rijswijk (WAG/Rij)and compare the results with control animals. -The density of GFAP, Cx30 and Cx43 positive astrocytes in per unite area were quantified in the CC of GAERS, WAG/Rij and control animals using immunohistochemistry and real-time quantitative polymerase chain reaction (RT-qPCR). The quantifications were made from three regions of CC; below the primary somatosensory (S1BF), below the motor (M1) and below the retrosplenial (RSG) cortices. oThe number GFAP, Cx30 and Cx43 immunopositive astrocytes showed heterogeneous distribution within the CC. The GFAP immunopositive astrocytes was significantly high in the S1BF region of the three strains. The immunopositive GFAP and Cx43 showed significant decrease in the S1BF and M1 regions in GAERS and WAG/Rij compared to control animals, however, an increase in the immunopositive Cx30 was observed in the same regions in both GAERS and WAG/Rij compared to control Wistar animals but the increase was significant for GAERS but not for WAG/Rij. The RT-qPCR analysis was corroborated by GFAP immunohistochemistry results. The different expression pattern of the two Cx's in the CC of the epileptic strains compared to control animals may indicate a compensatory response or maybe the cause of generalization of absence seizures.


Anticancer effects of myricetin derivatives in non-small cell lung cancer in vitro and in vivo.
Lung cancer is the most common cause of cancer-related deaths. Moreover, exploring efficient tumor-killing drugs is urgently needed. In our study, several derivative compounds of myricetin were synthesized and tested. Experiments on non-small cell lung cancer (NSCLC) showed that S4-2-2 (5,7-dimethoxy-3-(4-(methyl(1-(naphthalen-2-ylsulfonyl)piperidin-4-yl)amino)butoxy)-2-(3,4,5-trimethoxyphenyl)-4H-chromen-4-one) had the strongest effect on A549 cell inhibition across all compounds. Furthermore, S4-2-2-treated A549 cells were also suppressed when transplanted into immunodeficient mice. Particularly, we found that the migration and invasiveness of A549 cells became suppressed upon treatment with S4-2-2. Furthermore, the compound significantly induced cell apoptosis, but did not affect the cell cycle of A549 cells. Finally, we revealed that S4-2-2 inhibited the biological function of NSCLC cells by regulating the protein process in the endoplasmic reticulum, and then by inducing the expression of apoptosis-related proteins. Taken together, S4-2-2 was shown to act as a potential molecular inhibitor of A549 cells.


Determination of nine catecholamine metabolites and 5-hydroxyindolacetic acid in urine by capillary gas chromatography.
A method is described for the simultaneous determination of nine urinary acidic and alcoholic catecholamine metabolites and urinary 5-hydroxyindolacetic acid. Incubation of a urine sample in the presence of ascorbic acid, glucuronidase and acylase and subsequent extraction with ethyl acetate precedes derivatization to trimethylsilyl compounds, capillary gas chromatographic separation and flame-ionization detection. The automated dual injection procedure and the analytical characteristics of the proposed method are reported in detail. Special attention is paid to problems that occur in analysis on a routine basis.


Sleep in conduct-disordered adolescents--a polysomnographic and spectral power analysis study.
The aim of the present study was to characterize sleep in conduct-disordered adolescents using polysomnography and spectral power analysis. The two hypotheses were that conduct disorder would be associated with objective sleep problems, and that conduct disorder--as a precursor of adult antisocial personality disorder--would be associated with the same kind of abnormal sleep architecture, with both increased deep sleep and delta power, as previously reported in antisocial personality disorder. The patients consisted of 15 adolescents (age range 13-17 years, mean age 14.7 years) with histories of antisocial behavior so functionally impairing that they were ordered by child welfare to undergo a psychosocial evaluation in a closed social services ward. The healthy age-matched controls comprised 20 volunteers recruited with a newspaper advertisement. Opposite to earlier subjective sleep studies among conduct-disordered children, no significant differences in sleep parameters were observed between the two groups. The adolescents with conduct disorder slept a little bit longer, but the percentage amount of different sleep stages did not differ significantly between the two groups. Relative spectral power of sleep, delta power in particular, was similar in both groups, assessed in total sleep time as well as in first half of it. Different alternative explanations for these findings are discussed.


The Regulatory Role of Apelin on the Appetite and Growth of Common Carp (Cyprinus Carpio L.).
Apelin, a kind of active polypeptide, has many biological functions, such as promoting food intake, enhancing immunity, and regulating energy balance. In mammals, studies have indicated that apelin is involved in regulating food intake. However, there are relatively few studies about the regulatory effect of apelin on fish feeding, and the specific mechanism is not clear. Therefore, the purpose of this study was to preliminarily investigate the regulatory effects of apelin on key genes of feeding and growth in common carp (Cyprinus Carpio L.) through in vitro and in vivo experiments. In the present study, after incubation with different concentrations of Pyr-apelin-13 (0, 10, 100, and 1000 nM) in hypothalamic fragments, the expressions of Neuropeptide Y (NPY) and Agouti related peptide (AgRP) mRNA were significantly up-regulated at 12 and 3 h, respectively, and the significant down-regulation of Cocaine and amphetamine-related transcript (CART) mRNA expression was observed at 1 and 3 h. In vivo, after Pyr-apelin-13 oral administration (0, 1, 10, and 100 pmol/g), the orexin mRNA level in the hypothalamus of common carp was significantly increased at 1, 6, and 12 h, while CART/(Proopiomelanocortin) POMC mRNA levels in the hypothalamus of common carp were significantly down-regulated. Following incubation with different concentrations of Pyr-apelin-13 (0, 10, 100, and 1000 nM) in primary hepatocytes, GHR (Growth hormone receptor), IGF2 (Insulin-like growth factor 2), IGFBP2 (Insulin like growth factor binding protein 2), and IGFBP3 (Insulin like growth factor binding protein 3) mRNA levels were significantly increased at 3 h. In vivo, the levels of IGF1 (Insulin-like growth factor 1), IGF2, IGFBP2 (Insulin like growth factor binding protein 2), and IGFBP3 mRNA were significantly increased after the oral administration of Pyr-apelin-13 in the hepatopancreas, in a time and dose-dependent manner. These results support the hypothesis that Pyr-apelin-13 might regulate the feeding and growth of common carp through mediating the expressions of appetite- and growth-related genes. Overall, apelin, which is an orexigenic peptide, improves food intake and is involved in the growth of common carp.


[Obesity is rarely curable: individual concepts and therapy programs for children and adolescents].
Obesity in children has an increased risk to persist in adulthood. In most cases, obesity starts to develop in children at school age. Lower social economic status and migration background are severe risk factors for obesity. Additionally, genetic predisposition for the development of obesity plays an especially important role in children and adolescents. However, the individual cause for obesity is heterogeneous and complex. This is the reason why only a systematic analysis of individually existing problems is necessary for a differentiated and realistic planning of the treatment. Long-lasting therapy concepts need to be based on current available evidence.The treatment of childhood obesity should rely on multiprofessional lifestyle programs. An exception might be rare monogenic or syndromic forms of obesity, because defects within the central regulatory pathways of body weight regulation could be present. In general, a key component of the treatment strategy should include an improvement of nutrition, physical exercise and self-esteem combined with a reduction of stress. Moreover, the inclusion of parents into the treatment strategy has shown to be beneficial and necessary. Long-term follow-up studies on the development of associated comorbidities are rare. Unfortunately, patient groups at risk are currently not necessarily reached with available treatment programs.A multiprofessional analysis of individual problems and differentiated treatment planning with a participative approach (acknowledging the cultural background and involving members of the family) should lead to long-lasting improvement of the therapeutic outcome. The individual main treatment aim should also include - apart from the reduction of body weight - the improvement of associated comorbidities and quality of life, especially by avoiding any stigmatization. A health-promoting environment is desirable for this salutogenetic approach.


Antiinflammation Derived Suzuki-Coupled Fenbufens as COX-2 Inhibitors: Minilibrary Construction and Bioassay.
A small fenbufen library comprising 18 compounds was prepared via Suzuki Miyara coupling. The five-step preparations deliver 9-17% biphenyl compounds in total yield. These fenbufen analogs exert insignificant activity against the IL-1 release as well as inhibiting cyclooxygenase 2 considerably. Both the para-amino and para-hydroxy mono substituents display the most substantial COX-2 inhibition, particularly the latter one showing a comparable activity as celecoxib. The most COX-2 selective and bioactive disubstituted compound encompasses one electron-withdrawing methyl and one electron-donating fluoro groups in one arene. COX-2 is selective but not COX-2 to bioactive compounds that contain both two electron-withdrawing groups; disubstituted analogs with both resonance-formable electron-donating dihydroxy groups display high COX-2 activity but inferior COX-2 selectivity. In silico simulation and modeling for three COX-2 active-p-fluoro, p-hydroxy and p-amino-fenbufens show a preferable docking to COX-2 than COX-1. The most stabilization by the p-hydroxy fenbufen with COX-2 predicted by theoretical simulation is consistent with its prominent COX-2 inhibition resulting from experiments.


Connectome Analysis in an Individual with SETD1B -Related Neurodevelopmental Disorder and Epilepsy.
Causative variants in SETD1B , encoding a lysine-specific methyltransferase, have recently been associated with a neurodevelopmental phenotype encompassing intellectual disability, autistic features, pronounced language delay, and epilepsy. It has been noted that long-term and deep phenotype data are needed to further delineate this rare condition. In this study, we provide an in-depth clinical characterization with long-term follow-up and trio exome sequencing findings to describe one additional individual affected by SETD1B -related disorder. The diagnostic workup was complemented by a functional magnetic resonance imaging (fMRI) study. We report a 24-year-old male individual with an early-onset neurodevelopmental disorder with epilepsy due to the de novo missense variant c.5699A>G, p.(Tyr1900Cys) in SETD1B (NM_015048.1). He exhibited delayed speech development, autism spectrum disorder, and early-onset epilepsy with absence and generalized tonic-clonic seizures. Despite profoundly impaired communication skills, ongoing improvements regarding language production have been noted in adulthood. fMRI findings demonstrate abnormal language activation and resting-state connectivity structure. Our report expands the previously delineated phenotype of SETD1B -related disorder and provides novel insights into underlying disease mechanisms.


Using evaluative frameworks to examine the implementation outcomes of a cognitive behavioral therapy program for autistic students with anxiety within public school settings.
Cognitive behavioral therapy helps to treat anxiety symptoms in autistic youth, but it is difficult for families to access cognitive behavioral therapy in the community. Training school providers to deliver cognitive behavioral therapy may help autistic youth and their families to access these programs. Unfortunately, we do not know how cognitive behavioral therapy programs can be delivered by school providers and how these programs help the autistic students who access them. This study addressed this gap and was part of a larger study that looked at the effectiveness of Facing Your Fears-School-Based in 25 public schools. The study goals were to understand whether Facing Your Fears-School-Based helped students and the factors that made it easy or difficult to deliver Facing Your Fears-School-Based in schools. Thirty providers participated in interviews guided by the Reach, Effectiveness, Adoption, Implementation, and Maintenance framework. Participants shared information that fell into several major categories that included (1) delivering Facing Your Fears-School-Based to many different students; (2) the positive impact of Facing Your Fears-School-Based on students' school participation; and (3) plans to continue using Facing Your Fears-School-Based. School providers also shared that Facing Your Fears-School-Based was easy to use for non-mental health providers and reported adapting Facing Your Fears-School-Based to meet student needs. The results of this study suggest that Facing Your Fears-School-Based may help autistic students and highlight the importance of using mental health programs in schools that are flexible, able to be adapted, and that are able to be used by many different types of school providers.


Effects of exposure to low-level lead on spatial learning and memory and the expression of mGluR1, NMDA receptor in different developmental stages of rats.
The effect of different periods of lead exposure on deficits of learning and memory is still unclear. In this study, we conduct in vivo experiment to investigate the critical stages when lead induced neurotoxicity in rats and its underlying mechanisms in some critical stages. Rats were exposed to 0.2% mg/ml lead acetate solution via drinking water during gestation, lactation and ablactation periods. Behavior deficits were found in gestation and lactation. N-Methyl-d-aspartate (NMDA) receptor subunit 2A (NR2A) increased during gestation both in hippocampus and cerebral cortex compared to the control group; in all treatment groups NR2B decreased in hippocampus and in cerebral cortex during the lactation period. Meanwhile, in hippocampus metabotropic glutamate receptor 1 (mGluR1) decreased during gestation and lactation periods but increased during the ablactation period. These observations suggest that exposure to lead in gestation and lactation periods could cause neurobehavioral deficits which extend to adulthood, and lactation was a more sensitive period for lead exposure. Furthermore, the abnormal expression of NMDA receptor 2 (NMDAR 2) subunits and mGluR1 are likely to be associated with the impairment.


A new mixed micellar preparation for oral vitamin K prophylaxis: randomised controlled comparison with an intramuscular formulation in breast fed infants.
To compare a new oral preparation of vitamin K1 (Konakion MM) containing lecithin and glycocholic acid with a standard intramuscular (IM) preparation during the first 8 weeks of life in exclusively breast fed infants. Infants were randomised at birth to the IM group (1 mg vitamin K) or the oral group (2 mg given at birth and repeated at 7 and 30 days of life). Prothrombin time (INR), plasma vitamin K1, and PIVKA II (undercarboxylated prothrombin) were monitored at 14, 30, and 56 days of age. Seventy nine infants were randomised to the oral group and 77 to the IM group. Sixty seven infants in each group completed eight weeks of the study. Prothrombin times did not differ between the two groups. Mean (SD) plasma vitamin K1 values (in ng/ml) decreased in both groups over time, but were higher in the oral group at 14 and 56 days: 2.0 (1.6) v 1.3 (1.1) at 14 days; 0.5 (0.3) v 0.5 (0.7) at 30 days; and 0.5 (0.8) v 0.2 (0.2) at 56 days of life. PIVKA II was raised (> or = 0.1 AU/ml) in cord blood in 47% of the infants. By 14 days, only one infant in each group had a raised PIVKA II value and both of these initially had high concentrations of PIVKA II in cord blood. At 30 days, there were no raised PIVKA II values. At 56 days, there were no raised PIVKA II values in the oral group, although three infants in the IM group had raised values. Plasma vitamin K concentrations were at least equal or significantly higher in babies given oral vitamin K supplements compared with IM treated babies at the time points measured. Through the first 8 weeks of life, multiple doses of the new oral preparation maintain haemostasis and vitamin K status in breast fed infants at least equal to that of the intramuscular preparation.


Autologous minced cartilage repair for chondral and osteochondral lesions of the knee joint demonstrates good postoperative outcomes and low reoperation rates at minimum five-year follow-up.
Minced cartilage is a one-step, autologous procedure with promising short-term results. The aim of the present study was to evaluate mid-term results in a patient cohort with chondral and osteochondral lesions in the knee joint treated with minced cartilage. From 2015 through 2016, a total of 34 consecutive patients were treated with a single-step, autologous minced cartilage for knee chondral and osteochondral lesions. Numeric analogue scale (NAS) for pain and knee function were obtained prior to surgery and at 12, 24 and 60 months postoperatively. Secondary outcomes, including Lysholm score, Tegner activity score, and the International Knee Documentation Committee (IKDC) score, were recorded at final follow-up. MRI examinations of patients with unplanned radiological follow-up were analysed using the MOCART (Magnetic Resonance Observation of Cartilage Repair Tissue) score. A total of 28 patients (44.1% females, age at surgery: 29.5 ± 11.5 years) were available at a mean follow-up of 65.5 ± 4.1 months. Mean defect size was 3.5 ± 1.8 cm2. NAS for pain decreased from a median of 7 (range: 2-10) preoperatively to 2 (0-8) postoperatively. NAS knee function improved from a median of 7 (range: 2-10) to 3 (0-7) after five years, respectively. Satisfactory Lysholm (76.5 ± 12.5), IKDC (71.6 ± 14.8) and Tegner activity (4, range 3-9) scores were reported at final follow-up. Of all patients, 21(75%) and 19 (67.9%) reached or exceeded the PASS for the IKDC- and Lysholm score at final follow-up, respectively. The average overall MOCART 2.0 scores for all postoperatively performed MRIs (n = 23) was 62.3 ± 17.4. Four (14.2%) postoperative complications were directly linked to minced cartilage, one (3.5%) of which required revision surgery. One-step, autologous minced cartilage repair of chondral and osteochondral lesions of the knee without the necessity for subchondral bone treatment demonstrated good patient-reported outcomes, low complication rates, and graft longevity at mid-term follow-up. Minced cartilage represents a viable treatment option to more traditional cartilage repair techniques even in mid-term. Level III.


Electrophysiological and dye-coupling studies on secretory, myoepithelial and duct cells in human eccrine sweat glands.
1. Electrophysiological properties and dye-coupling status of secretory, myoepithelial and coiled duct cells in isolated human eccrine sweat glands have been assessed by single-micro-electrode studies and intracellular micro-iontophoresis of the fluorescent naphthalimide dye Lucifer Yellow CH (molecular weight 457). Treated glands were embedded in LKB HistoResin and examined by transmission fluorescence microscopy, first as wholemounts and then as 5 microns serial sections. Sections positive for Lucifer Yellow were photographed and then stained with Toluidine Blue for observation by conventional microscopy to confirm the site of penetration. 2. Out of forty-five successful micro-iontophoreses, three were confirmed in secretory cells, twelve in myoepithelial cells and thirty in cells of the coiled duct wall. The latter were identified as the most penetrable in the coiled part of the isolated human eccrine sweat gland. 3. Of the three secretory cells labelled (resting potentials -40, -52 and -63 mV), all demonstrated dye coupling to neighbouring secretory cells although in one case this was found to be selective. Not every secretory cell was involved in coupling. No fluorescent label spread to the myoepithelial cells which form a network on the basal surface of the secretory tubule. 4. When myoepithelial cells were penetrated, they demonstrated dye coupling to neighbouring myoepithelial cells but not to secretory cells with which they also made contact. Basal resting potentials of -35 to -65 mV were recorded (mean = -52 mV, S.E. of mean = +/- 2.4 mV, n = 12) and, in eight out of the twelve cells penetrated and labelled, spontaneous depolarizing transients were also observed whose amplitude but not frequency increased with increasing membrane polarization. Administration of acetylcholine to produce a final concentration of 10(-6) to 10(-7) M produced either depolarization or micro-electrode dislodgement. 5. Of the thirty cells labelled in the coiled duct, twenty-six showed obvious dye spread to neighbouring cells in both layers of the wall. There was no relation between dye-coupling status and basal resting potential which lay in the range -40 to -82 mV (mean = -60 mV, S.E. of mean = +/- 2.4 mV, n = 30). Repeated doses of acetylcholine produced either no response from cells in this range or depolarization in cells with resting potentials more negative than -70 mV and hyperpolarization in cells with resting potentials more positive than -70 mV. In addition, there was a biphasic response, depolarization followed by hyperpolarization in a cell of resting potential -67 mV.(ABSTRACT TRUNCATED AT 400 WORDS)


Immune escape as a fundamental trait of cancer: focus on IDO.
Immune escape is a critical gateway to malignancy. The emergence of this fundamental trait of cancer represents the defeat of immune surveillance, a potent, multi-armed and essential mode of cancer suppression that may influence the ultimate clinical impact of an early stage tumor. Indeed, immune escape may be a central modifier of clinical outcomes, by affecting tumor dormancy versus progression, licensing invasion and metastasis and impacting therapeutic response. Although relatively little studied until recently, immune suppression and escape in tumors are now hot areas with clinical translation of several new therapeutic agents already under way. The interconnections between signaling pathways that control immune escape and those that control proliferation, senescence, apoptosis, metabolic alterations, angiogenesis, invasion and metastasis remain virtually unexplored, offering rich new areas for investigation. Here, an overview of this area is provided with a focus on the tryptophan catabolic enzyme indoleamine 2,3-dioxygenase (IDO) and its recently discovered relative IDO2 that are implicated in suppressing T-cell immunity in normal and pathological settings including cancer. Emerging evidence suggests that during cancer progression activation of the IDO pathway might act as a preferred nodal modifier pathway for immune escape, for example analogous to the PI3K pathway for survival or the VEGF pathway for angiogenesis. Small molecule inhibitors of IDO and IDO2 heighten chemotherapeutic efficacy in mouse models of cancer in a nontoxic fashion and an initial lead compound entered phase I clinical trials in late 2007. New modalities in this area offer promising ways to broaden the combinatorial attack on advanced cancers, where immune escape mechanisms likely provide pivotal support.


Inducibility of chromosomal aberrations by metal compounds in cultured mammalian cells.
Metal compounds were tested for their ability to induce chromosomal aberrations in cultured mammalian cells. Chromosomal aberrations were induced by the application of some Cr, Mn and Ni compounds. Among 6-valent Cr compounds, K2Cr2O7 and CrO3 induced high levels of aberrations, at rates which were similar for Cr-equivalent doses. The perchromate compounds were more efficient in producing chromosomal aberrations than was a chromate compound, K2CrO4. A 3-valent Cr compound, Cr2(SO4)3, was less toxic and failed to induce a demonstrable increase in chromosomal aberrations. KMnO4 induced aberrations, but at a low rate. As to Ni compounds, NiCl2 and (CH3COO)2Ni induced few aberrations. Administration of K2Ni(CN)4 induced only gaps. NiS induced a low but definite increase in chromosomal aberrations. The rate of these aberrations increased with an increase in treatment time from 24 to 48 h, indicating a time-dependent increase in the hereditable toxicity of metal compounds. CdCl2 and HgCl2 were somewhat toxic, but failed to induce chromosomal aberrations in the present study.


The drinking water disinfection performances and mechanisms of UVA-LEDs promoted by electrolysis.
In this study, the UVA (Ultraviolet A) drinking water disinfection was promoted by electrolysis. The influences of the UVA, electrolysis current, bubbling and temperature were investigated. The disinfection mechanisms and bacterial reactivation had been studied. The results revealed that the treatment time needed to reach the DL (detection limit, about 5.4 log removal) was shortened from 180 to 80 min by the electrolysis. The total electricity consumption decreased from about 126-57.0 kJ/L. Compared with increasing the UVA irradiation, increasing the electrolysis current in a certain range was more preferred to improve the disinfection rate. Oxygen bubbling or higher temperature could enhance the E. coli inactivation. The quenching experiment and EPR (Electron paramagnetic resonance) detection confirmed that ROSs (1O2, ·O2- and ·OH) played important roles for the disinfection. Compared with the treatment with UVA alone, the cell membrane damage was more severe by the promoting method. In addition to the dramatically reduced enzyme activity, the synergistic process degraded most of the bacterial genomic DNA, and the bacteria were completely killed. Therefore, hybrid with electrolysis is a better way for the application of the UVA-LED disinfection.


Spongiform encephalopathy with extensive involvement of white matter.
We report a proven case of CJD with prolonged clinical course and white matter involvement which consisted of severe, widespread myelin damage in the forebrain and cerebellum but with sparing of the internal capsule. Histologically, there was status spongiosus of the involved white matter with axonal loss, proliferation of hypertrophic astrocytes and scattered and perivascular foamy macrophages. White matter lesions have not been considered to be a significant feature of CJD. In the last few years, however, a few cases have been described with prominent degeneration of cerebral white matter. We believe that our findings corroborate the existence of an entity that at present may only be defined as spongiform panencephalopathy.


The entry of gossypol across the blood-testis barrier in rats.
This paper presents the experimental data about how different formulations of gossypol enter the blood-testis barrier following intravenous administration in acute experiments on rats. It was found that gossypol encapsulated by liposomes crossed the blood-testis barrier more readily than free gossypol, without affecting its pharmacokinetic pattern in the circulating blood, suggesting that liposomes may be useful as drug carriers and may facilitate the entry of encapsulated gossypol into the seminiferous tubule from the blood.


Electron counting detectors in scanning transmission electron microscopy via hardware signal processing.
Transmission electron microscopy is a pivotal instrument in materials and biological sciences due to its ability to provide local structural and spectroscopic information on a wide range of materials. However, the electron detectors used in scanning transmission electron microscopy are often unable to provide quantified information, that is the number of electrons impacting the detector, without exhaustive calibration and processing. This results in arbitrary signal values with slow response times that cannot be used for quantification or comparison to simulations. Here we demonstrate and optimise a hardware signal processing approach to augment electron detectors to perform single electron counting.


Modelling induction of a rotor in cardiac muscle by perpendicular electric shocks.
A strong, properly timed shock applied perpendicularly to a propagating wavefront causes a rotor in the canine myocardium. Experimental data indicate that the induction of this rotor relies on the shock exciting tissue away from the electrodes. The computational study reproduced such direct excitation in a two-dimensional model of a 2.7 x 3 cm sheet of cardiac muscle. The model used experimentally measured extracellular potentials to represent 100 and 150 V shocks delivered through extracellular electrodes. The shock-induced transmembrane potential was computed according to two mechanisms, the activating function and the unit-bundle sawtooth potential. The overall process leading to initiation of a rotor was the same in model and experiment. For the 100 V shock, the directly excited region extended 2.26 cm away from the electrode; the centre of the rotor ('critical point') was 1.28 cm away, where the electric field Ecr was 4.54 Vcm(-1). Increasing the shock strength to 150 V moved the critical point 1.02 cm further and decreased Ecr by 0.39 Vcm(-1). The results are comparable with experimental data. The model suggests that the unit-bundle sawtooth is responsible for the creation of the directly excited region, and the activating function is behind the dependence of Ecr on shock strength.


Expression Patterns of Coagulation Factor XIII Subunit A on Leukemic Lymphoblasts Correlate with Clinical Outcome and Genetic Subtypes in Childhood B-cell Progenitor Acute Lymphoblastic Leukemia.
Based on previous retrospective results, we investigated the association of coagulation FXIII subunit A (FXIII-A) expression pattern on survival and correlations with known prognostic factors of B-cell progenitor (BCP) childhood acute lymphoblastic leukemia (ALL) as a pilot study of the prospective multi-center BFM ALL-IC 2009 clinical trial. The study included four national centers (n = 408). Immunophenotyping by flow cytometry and cytogenetic analysis were performed by standard methods. Copy number alteration was studied in a subset of patients (n = 59). Survival rates were estimated by Kaplan-Meier analysis. Correlations between FXIII-A expression patterns and risk factors were investigated with Cox and logistic regression models. Three different patterns of FXIII-A expression were observed: negative (<20%), dim (20-79%), and bright (≥80%). The FXIII-A dim expression group had significantly higher 5-year event-free survival (EFS) (93%) than the FXIII-A negative (70%) and FXIII-A bright (61%) groups. Distribution of intermediate genetic risk categories and the "B-other" genetic subgroup differed significantly between the FXIII-A positive and negative groups. Multivariate logistic regression confirmed independent association between the FXIII-A negative expression characteristics and the prevalence of intermediate genetic risk group. FXIII-A negativity is associated with dismal survival in children with BCP-ALL and is an indicator for the presence of unfavorable genetic alterations.


Autologous blood injection for treatment of lateral epicondylosis: A meta-analysis of randomized controlled trials.
To appraise existing evidence of autologous blood injection in treating lateral epicondylosis. Meta-analysis of randomized controlled trials. A comprehensive search of the PubMed, Cochrane, SCOPUS, and CINAHL databases was performed to identify randomized controlled trials that reported the efficacy of autologous blood injection in treating lateral epicondylosis. The selected studies were subjected to a meta-analysis and risk of bias assessment. Patients with lateral epicondylosis. Pain-related measurement in each selected randomized controlled trial was pooled into meta-analysis. Nine randomized controlled trials were included in the analysis. The results of the meta-analysis including the pain scores indicated that autologous blood injection is more effective compared with corticosteroid injection (standard mean difference: -0.75; 95% confidence interval: -1.14 to -0.37) but not more effective compared with platelet-rich plasma injection (standard mean difference: 0.09; 95% confidence interval: -0.66 to 0.84). The risk of bias assessment indicated that all the included trials exhibited a moderate to high risk of bias. Autologous blood injection is more effective than corticosteroid injection but not more effective than platelet-rich plasma injection in treating lateral epicondylosis. However, this evidence is limited by the potential risk of bias.


Inflammatory and immunological responses in skin and peripheral lymph of sheep following intracutaneous injection of Staphylococcus aureus.
The cellular response within lesions and in draining lymph was examined in sheep following a primary intracutaneous injection of live or killed S. aureus. Microscopic examination of sections from live S. aureus lesions (12, 24, 48, and 96 h following vaccination) revealed a high ratio of neutrophils to macrophages at all times. This ratio was initially high following inoculation of killed S. aureus but decreased steadily at successive sampling times. Representative sections from lesions were subjected to indirect immunofluorescent staining to identify IgM-, IgG1-, and IgG2-containing cells. The ratio of IgG2- to IgG1-containing cells in lesions produced following live S. aureus vaccination was significantly greater than the ratio in lesions produced by killed staphylococci. Lesions induced by live S. aureus recruited significantly greater numbers of 51Cr-labeled allogeneic neutrophils from blood than did lesions induced by killed S. aureus. During the first 6 h this difference was approx. 20-fold. The volume of lymph and the number of leukocytes draining live S. aureus lesions was considerably greater than from lesions produced by killed staphylococci. The proportion of neutrophils in lymph draining both types of lesions increased markedly during the first two days of the response but was observed to be greater and remained higher for a longer period of time in lymph draining vaccine lesions produced following injection of live staphylococci. The increase in proportion of neutrophils in lymph was accompanied by a concomitant decrease in proportion of lymphocytes and macrophages. No immunoglobulin-containing cells or anti-staphylococcal antibody production was detected in lymph draining either type of lesion. These differences in inflammatory responses may contribute to the documented differences in immune responses to live and killed staphylococcal vaccines.


The adsorption and absorption kinetics of BDE-47 by Chlorella sp. and the role of extracellular polymer substances influenced by environmental factors.
Microalgae act as the entrance of polybrominated diphenyl ethers (PBDEs) from abiotic to biotic environments, which controlled the environmental fate of PBDEs in aquatic environments. Combing with typical coastal environmental characteristics including extracellular polymer substances (EPS) enrichment, light limitation and nitrogen starvation, the changes of adsorption and absorption kinetics of BDE-47 by Chlorella sp. and the role of EPS therein were investigated. The results quantified the adsorption and absorption kinetics of BDE-47 by Chlorella sp. cells and fitted it by the Lagergren pseudo first order model. Furthermore, we found the adsorption and absorption kinetics could be changed by the above mentioned environmental factors. To be specific, the total BDE-47 adsorption amounts per microalgal cell were increased as the increase of ambient EPS (proteins or carbohydrates), attributing to the increase of soluble (SL)-EPS contents; increased total BDE-47 adsorption amounts but decreased absorption rates were found under light limitation and nitrogen starvation, which were attributed to increased bound (B)-EPS contents and protein/carbohydrates (P/C) ratios therein, respectively. Therefore, our study elucidated the adsorption and absorption kinetics of PBDEs by microalgae could be influenced by ambient environmental changes, clarified the roles of SL-EPS, B-EPS contents and P/C ratios, providing a solid basis for evaluating the environmental fate of PBDEs in the marine environments.


Repeated intracoronary beta radiation for recurrent in-stent restenosis.
More than 70% of percutaneous coronary interventions are followed by a stent implantation. In-stent restenosis still occurs in 20-30% of patients and remains a therapeutic challenge. At present only vascular brachytherapy has been shown to be an effective treatment option. We report here one case of recurrent in-stent restenosis after vascular brachytherapy that was successfully treated by a second beta radiation treatment.


Large Stokes shift induced by intramolcular charge transfer in N,O-chelated naphthyridine-BF2 complexes.
Novel N,O-chelated naphthyridine-BF(2) complexes with push-pull structures have been synthesized and characterized. Spectral investigations on these complexes reveal that photoinduced intramolecular charge transfer occurs and results in a large Stokes shift, which is further supported by density functional theory based theoretical calculations.


Assessment of heavy metal accumulation potential of aquatic plants for bioindication and bioremediation of aquatic environment.
This study is devoted to assessing the potential for accumulating heavy metals (Fe, Mn, Cu, Zn) of nine species of aquatic and coastal plants growing in the water area of the Okhta River (St. Petersburg, Russian Federation). The list of studied species included the following: hornwort (Ceratophyllum demersum L.), water soldier (Stratiotes aloides L.), yellow water-lily (Nuphar lutea (L.) Sm.), arrowhead (Sagittaria sagittifolia L.), broadleaf cattail (Typha latifolia L.), water arum (Calla palustris L.), acute sedge (Carex acuta L.), reed canary grass (Phalaris arundinacea L.), and meadowsweet (Filipendula ulmaria (L.) Maxim.). The values of the bioconcentration factor for particular plant tissues were calculated. It has been established that the concentrations of metals in plant tissues can be thousands and tens of thousands of times higher than their content in water. At the same time, the ability of plants to accumulate metals varies greatly and depends on a number of factors, such as environmental conditions, the growing season, the type and ecological group of the plant.The study also attempts to determine the spatial and temporal dynamics of the content of heavy metals in higher aquatic plants. The main difficulties in data interpretation are indicated. The data obtained show the different significance of the study macrophyte species both for bioindication purposes and for use in wastewater treatment. The possibility of using such species as Sagittaria sagittifolia L., Typha latifolia L., Ceratophyllum demersum L., and Calla palustris L. for bioremediation is substantiated.


Chin Microgenia: A Clinical Comparative Study.
The aesthetic relevance of the chin and its relatively simple correction through different approaches make genioplasty one of the most performed aesthetic procedures of the face. Sliding genioplasty is extremely rewarding, particularly when performed as an adjunction to rhinoplasty, rhytidectomy, or jaw surgery. In the scientific literature, many different surgical techniques are described, but the biological implications and the economical impact can shape the surgeon's decision on which can be the best treatment: surgical correction with osteotomy, chin implants, or with fillers. The authors propose a decision making protocol for correcting chin microgenia based on a revision of 345 treated cases. A retrospective review of 345 cases of chin microgenia was undertaken to understand the proper preoperative assessment and therapeutic planning. A total of 135 patients were treated with surgical sliding genioplasty (group A): 60 patients (group B) have been grafted with alloplastic implants and the remaining 150 patients (group C) with hyaluronic acid. We recorded clinical indications, complications, and long-term aesthetic results at 3-year follow-up. The analysis of the results based on the entity of the chin's sagittal defect, the chin soft-tissue thickness, the patient's age, and self-judgment allows for simplified treatment planning for sagittal chin deformities showing a greater predictability and a more stable long-term aesthetic result regarding sliding genioplasty compared to alloplastic implant placement and fillers. Our proposal for a simple and versatile protocol of chin microgenia aims to simplify the therapeutic indications for a predictable and a stable long-term aesthetic result. This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to Table of Contents or the online Instructions to Authors www.springer.com/00266 .


Turnera diffusa extract attenuates profibrotic, extracellular matrix and mitochondrial markers in activated human hepatic stellate cells (HSC).
Hepatic fibrosis is characterized by the accumulation of extracellular matrix which includes the accumulation of α-smooth muscle actin (α-SMA), collagen type I (COL1α1), as well as remodeling induced by metalloproteinases and tissue inhibitor of metalloproteinase (TIMPs), where hepatic stellate cells (HSCs) play a central role. In addition, the transcription factor SNAI1 (which participates in epithelial-mesenchymal transition, EMT) and mitofusin 2 (MFN2, a mitochondrial marker) plays an important role in chronic liver disease. Turnera diffusa (TD), a Mexican endemic plant, has been shown to possess antioxidant and hepatoprotective activity in vitro. We treated human HSC (LX2 cells) with a methanolic extract of Turnera diffusa (METD) to evaluate the mechanism involved in its hepatoprotective effect measured as fibrosis modulation, EMT, and mitochondrial markers. HSC LX-2 cells were treated with METD (100 and 200ng/mL) alone or combined with TGF-β (10ng/mL) at different time points (24, 48, and 72h). α-SMA, COL1α1, MMP2, TIMP1, SNAI1, and MFN2 mRNAs and protein levels were determined by real-time quantitative PCR and Western Blot analysis. We found that METD decreases COL1α1-mRNA, α-SMA, and TIMP1 protein expression in LX2 cells treated with and TGF-β. This treatment also decreases MFN2 and TIMP1 protein expression and induces overexpression of MMP2-mRNA. Our results suggest that a methanolic extract of Turnera diffusa is associated with an antifibrotic effect by decreasing profibrotic and mitochondrial markers together with the possible induction of apoptosis through SNAI1 expression in activated HSC cells.


Quantitative assessment of the effect of hepatic lipase gene polymorphism on the risk of coronary heart disease.
The human hepatic lipase (LIPC) is a glycoprotein member of the lipase superfamily that has attracted considerable attention as a candidate gene for coronary heart disease (CHD) based on its enzyme function as a key factor in lipoprotein catabolism pathways. In the past decade, a number of case-control studies have been carried out to investigate the relationship between the LIPC polymorphisms and CHD. However, studies on the association between LIPC polymorphisms and CHD remain conflicting. To derive a more precise estimation of the relationship, a meta-analysis of 11,906 cases and 13,273 controls from 18 published case-control studies was performed. Overall, the summary odds ratio of CHD was 0.87 (95% confidence interval: 0.66-1.15) and 1.03 (95% confidence interval: 0.98-1.07) for LIPC -250A and -514T alleles, respectively. No significant results were observed in heterozygous and homozygous when compared with wild genotype for these polymorphisms. In the stratified analyses according to ethnicity, source of controls, no evidence of any gene-disease association was obtained. Our result suggest that the G-250A, C-514T polymorphisms of LIPC gene are not associated with CHD susceptibility.


Textbook outcome in the surgical treatment of liver hydatid cyst.
Textbook outcome is a composite measure used in surgery to define the ideal postoperative period and to assess the quality of care. The aim of this study was to analyze the incidence of textbook outcome and the factors independently associated with its achievement following surgical treatment of liver hydatid cysts. Retrospective cohort study of patients operated on for liver hydatid cysts between January 2006 and December 2021. Textbook outcome was achieved when all the following criteria were fulfilled: no mortality within 90 days, no major complications within 90 days, no hospital readmission within 90 days, and no prolonged hospital stay. Univariable and multivariable analyses were performed to identify factors associated with textbook outcome. During the study period, 296 patients underwent surgery. Textbook outcome was recorded in 65.9% (195/296). Female gender (odds ratio 2.02; P = .010), noncomplicated cyst (odds ratio 3.97, P < .001), and radical surgery (odds ratio 2.26, P = .003) were the variables associated with a higher probability of achieving textbook outcome. Textbook outcome may be a useful measure to assess the variations in surgical management between different centers, and to improve quality of care after liver hydatid cysts resection.


High prevalence of mutations in LCAT in patients with low HDL cholesterol levels in The Netherlands: identification and characterization of eight novel mutations.
Lecithin:cholesterol acyltransferase (LCAT) is crucial to the maturation of high-density lipoprotein (HDL). Homozygosity for LCAT mutations underlies rare disorders characterized by HDL-cholesterol (HDL-c) deficiency while heterozygotes have half normal HDL-c levels. We studied the prevalence of LCAT mutations in referred patients with low HDL-c to better understand the molecular basis of low HDL-c in our patients. LCAT was sequenced in 98 patients referred for HDL-c <5th percentile and in four patients referred for low HDL-c and corneal opacities. LCAT mutations were highly prevalent: in 28 of the 98 participants (29%), heterozygosity for nonsynonymous mutations was identified while 18 patients carried the same mutation (p.T147I). The four patients with corneal opacity were compound heterozygotes. All previously identified mutations are documented to cause loss of catalytic activity. Nine novel mutations-c.402G>T (p.E134D), c.403T>A (p.Y135N), c.964C>T (p.R322C), c.296G>C (p.W99S), c.736G>T (p.V246F), c.802C>T (p.R268C), c.945G>A (p.W315X), c.1012C>T (p.L338F), and c.1039C>T (p.R347C)--were shown to be functional through in vitro characterization. The effect of several mutations on the core protein structure was studied by a three-dimensional (3D) model. Unlike previous reports, functional mutations in LCAT were found in 29% of patients with low HDL-c, thus constituting a common cause of low HDL-c in referred patients in The Netherlands.


Amygdala-kindling as a model for chronic efficacy studies on antiepileptic drugs: experiments with carbamazepine.
The anticonvulsant properties of carbamazepine were evaluated after single and multiple doses in the kindled amygdaloid seizure model in rats. Carbamazepine significantly reduced severity and duration of motor seizures and the duration of afterdischarges recorded from the stimulated amygdala after a single dose of 30 mg/kg (i.p.), but tolerance developed during chronic treatment of rats with three times daily intraperitoneal injection of 30 mg/kg of the drug for 2 weeks. Tolerance was also observed with respect to central side-effects (ataxia, muscle relaxation) of the treatment. Determinations of plasma levels confirmed previous studies in rats and other species in that concentrations of carbamazepine declined during chronic treatment, which may be related to autoinduction of the metabolism of the drug. Indeed, carbamazepine-10,11-epoxide, the major active metabolite of carbamazepine, could not be quantified in the plasma after the first dose but accumulated during chronic treatment, indicating an enhanced metabolism of the parent drug. As a consequence of decreasing concentrations of carbamazepine but increasing levels of the epoxide, the sum of concentrations of both compounds after 1 or 2 weeks of treatment did not differ from the concentration of unchanged carbamazepine initially determined. Determinations of the acute anticonvulsant potency in kindled rats indicated that carbamazepine-10,11-epoxide was about half as potent as the parent drug. In view of the anticonvulsant activity of carbamazepine-10,11-epoxide, the loss of anticonvulsant efficacy observed during chronic treatment with carbamazepine cannot be related solely to metabolic tolerance, i.e. due to enhanced metabolism of drug, but should also relate to functional tolerance, i.e. induced by central adaptation to the long-term presence of the drug and its metabolite.


Progression from preplus to plus disease in the Telemedicine Approaches to Evaluating Acute-Phase Retinopathy of Prematurity (e-ROP) Study: incidence, timing, and predictors.
To determine the incidence of and timing and predictors for progression from pre-plus to plus disease, based on evaluation of images. Two trained readers independently evaluated posterior pole images of infants from 13 North American centers for pre-plus/plus disease, stage, and zone of retinopathy of prematurity (ROP). Discrepancies between readers were adjudicated. To be eligible for analysis, eyes had to have at least two imaging sessions, the earlier one with pre-plus disease. Of 681 eyes of 444 infants with pre-plus first detected at mean postmenstrual age (PMA) of 35.5 ± 2.1 weeks, 54 (7.9%) progressed to plus disease at a mean PMA of 37.6 ± 2.4 weeks with the mean interval for progression of 2.7 weeks (range, 0.4-8.9 weeks). Progression rate was higher for eyes with larger number of quadrants of pre-plus (44% for eyes with four quadrants vs 4% with one quadrant [P < 0.0001]), earlier PMA with pre-plus (18% for 32 weeks' PMA vs 3% for PMA of >37 weeks [P = 0.02]), higher ROP stage (12% for stage 3, 2.5% for no ROP [P < 0.0001]), lower ROP zone (24% for zone I, 6% for zone II or no ROP [P < 0.0001]) at the time of first pre-plus detection. Based on image evaluation, 8% of eyes progressed from pre-plus to plus disease at a mean interval of 3 weeks. Pre-plus in multiple quadrants, higher stages of ROP, and lower zones of ROP were associated with higher risk of progression. Image evaluation for pre-plus may help in the identification of high-risk eyes for developing plus disease.


Dose-response effects of an antimicrobial peptide, a cecropin hybrid, on growth performance, nutrient utilisation, bacterial counts in the digesta and intestinal morphology in broilers.
The aim of the present study was to evaluate the feasibility of an antimicrobial peptide, cecropin A(1-11)-D(12-37)-Asn (CADN), as an alternative to antibiotic growth promoter (AGP) in poultry diets. A total of 1500 14-d-old indigenous male chickens (222 (sd 13) g) were randomly allocated to five groups with five replicate cages of sixty birds each, and fed ad libitum five grower diets and subsequently five finisher diets for 14 d each. The diets were made up by supplementing their basal diets with a CADN liquid sample (CADNL) at 0, 2, 4, 6 and 8 ml/kg, respectively. During the feeding period, a metabolic experiment was carried out to determine the apparent digestibility of diethyl ether extract, nitrogen retention and apparent metabolisable energy of the diet sample fed to each cage of chicks. At the end of the feeding experiment, one chick from each cage was killed for bacteriological, light microscopic and scanning electron microscopic examination of the intestinal villi. CADN had a negative linear, positive quadratic and negative linear effect on feed intake (F), weight gain (G) and feed:gain ratio (F:G), respectively, for the growers; it had a quadratic effect on F, G or F:G for the finishers; it increased nutrient utilisation for both growers and finishers; it decreased aerobic bacterial counts in both jejunal and caecal digesta in a dose-dependent manner; it enhanced intestinal villus heights in a dose-dependent manner and made the duodenum villi of the CADNL8 group at 42 d appear as a netted leaf-like structure. CADN is therefore a possible alternative to AGP in broiler feeds.


An overview of magnetic ionic liquids: From synthetic strategies to applications in microextraction techniques.
Remarkable progress has been achieved in the application of magnetic ionic liquids in microextraction-based procedures. These materials exhibit unique physicochemical properties of ionic liquids featuring additional responses to magnetic fields by incorporating a paramagnetic component within the chemical structure. This intriguing property can open new horizons in analytical extractions because the solvent manipulation is facilitated. Moreover, the tunable chemical structures of magnetic ionic liquids also allow for task-specific extractions that can significantly increase the method selectivity. This review aimed at providing an up-to-date overview of articles involving synthesis, physicochemical properties, and applications of magnetic ionic liquids highlighting recent developments and configurations. Moreover, a section containing critical evaluation and future trends in magnetic ionic liquid-based extractions is included.


Time analysis of corneal endothelial cell density after cataract extraction.
Serial endothelial photographs were taken preoperatively and postoperatively in 200 eyes; 111 eyes contained a Rayner iris clip lens, 54 eyes contained a Fyodorov Sputnik lens, and 35 eyes had no lens. Central endothelial cell density was changed in all instances, with counts in implanted eyes declining 25 to 30%, and in nonimplanted eyes 10 to 15%. In both instances, the decline essentially ceased at about three months. The cause of the greater decline in implanted eyes appeared to be mechanical and subsequent cell loss after the 90-day period was virtually equal for the two groups. Methods that may be used to alter the difference in cell density occurring with implantation are best analyzed by using the 90-day period data for comparison.


Primary malignant melanoma of the esophagus.
The case of a 54-year-old man with primary malignant melanoma of the esophagus is described. The results of morphologic examination are presented, and the histogenesis of the tumor and the classification of melanomas are briefly discussed. Histologic examination suggested that this was a case of primary malignant melanoma of the esophagus arising from melanosis of the esophageal mucosa. Like melanomas from the oral mucosa, vagina, anus, and vulva, the tumor had lentiginous radial components.


The NDUFB6 subunit of the mitochondrial respiratory chain complex I is required for electron transfer activity: a proof of principle study on stable and controlled RNA interference in human cell lines.
Molecular bases of inherited deficiencies of mitochondrial respiratory chain complex I are still unknown in a high proportion of patients. Among 45 subunits making up this large complex, more than half has unknown function(s). Understanding the function of these subunits would contribute to our knowledge on mitochondrial physiology but might also reveal that some of these subunits are not required for the catalytic activity of the complex. A direct consequence of this finding would be the reduction of the number of candidate genes to be sequenced in patients with decreased complex I activity. In this study, we tested two different methods to stably extinct complex I subunits in cultured cells. We first found that lentivirus-mediated shRNA expression frequently resulted in the unpredicted extinction of additional gene(s) beside targeted ones. This can be ascribed to uncontrolled genetic material insertions in the genome of the host cell. This approach thus appeared inappropriate to study unknown functions of a gene. Next, we found it possible to specifically extinct a CI subunit gene by direct insertion of a miR targeting CI subunits in a Flp site (HEK293 Flp-In cells). By using this strategy we unambiguously demonstrated that the NDUFB6 subunit is required for complex I activity, and defined conditions suitable to undertake a systematic and stable extinction of the different supernumerary subunits in human cells.


A novel needle electrode for intraoperative fourth cranial nerve neurophysiological mapping.
Trochlear nerve (CN-IV) mapping method has not been confirmed to date. The compound muscle action potential (CMAP) of CN-IV cannot be recorded because of the low mapping sensitivity and anatomical characteristics of the superior oblique muscle (SOM). The aim of this study was to evaluate the effectiveness of a novel needle electrode (NNE), for the intraoperative mapping of CN-IV. The NNEs were inserted in the target extraocular muscles in 19 patients. We compared the CMAP amplitude of the NNE with that of the conventional needle electrode (CNE). Furthermore, we investigated the dissimilarity between the CMAP of the CN-IV and other extraocular cranial nerves (ECNs) and the correlation between the readings of the CN-IV mapping and its postoperative functional outcome. The CMAP of CN-IV has been measured in nine patients (47.4%). The CMAP of CN-IV was distinguishable from other ECNs. The CMAP of the NNE was found to be three times higher than that of the CNE. Although the NNE has shown the potential to record the CN-IV's CMAP, 4 cases ended up having a CN-IV postoperative dysfunction. For the first time, we confirmed the possibility of intraoperative mapping the CN-IV using an NNE inserted into the SOM. The NNE can also be useful for other neurophysiological monitoring methods.


Fundamentals of power spectra of extracellular potentials produced by a skeletal muscle fibre of finite length. Part I: Effect of fibre anatomy.
To provide a thorough understanding of the changes in the power spectrum of electromyographic (EMG) signals, the formation of the power spectrum (PS) of extracellular potentials (EPs) produced by a skeletal muscle fibre of finite length was analysed. It was shown that, as in the case of an infinite fibre, the PS could be represented as the product of power spectra of the input signal (the first temporal derivative of the intracellular action potential, IAP) and of the impulse response (IR) of the fibre of finite length as a system of EP generation. The interrelations between the two multipliers determine the sensitivity of the EP power spectrum to alterations in parameters. The anatomical parameters of the fibre (length, depth, position of the end-plate in respect of the fibre ends) affect the EP power spectrum through IR power spectrum. Variations of the EP characteristic frequencies along the fibre length as well as oscillations in the PS are intrinsic properties of the fibre of finite length.


Diagnosis of inflammatory bowel disease-Asian perspectives: the results of a multinational web-based survey in the 8th Asian Organization for Crohn's and Colitis meeting.
Inflammatory bowel disease (IBD) is no longer a rare disease in Asia, thus it needs to prepare recommendations relevant to Asian patients. This study aimed to identify disparities in the process of the diagnosis of IBD in Asian countries/regions. In line with the 2020 Asian Organization for Crohn's and Colitis annual meeting, a multinational web-based survey about Asian physicians' perspectives on IBD was conducted. A total of 384 Asian physicians (99 in China, 93 in Japan, 110 in Korea, and 82 in other Asian countries/regions) treating IBD patients from 24 countries/regions responded to the survey. Most respondents were gastroenterologists working in an academic teaching hospital. About half of them had more than 10 years of clinical experience in caring for patients with IBD. The European Crohn's Colitis Organisation guideline was used most commonly for the diagnosis of IBD except for Japanese physicians who preferred their own national guideline. The Mayo score and Crohn's Disease Activity Index were the most commonly used activity scoring systems for ulcerative colitis and Crohn's disease, respectively. Endoscopy, not surprisingly, was the main investigation in assessing the extent and activity of IBD. On the other hand, there were disparities across countries/regions with regard to the favored modalities of small bowel and perianal evaluation of Crohn's disease, as well as the use of serologic markers. Results of the present survey revealed practical behaviors of Asian physicians in the diagnosis of IBD. Investigating the reasons for different diagnostic approaches among countries/regions might help us develop Asian guidelines further.


Leveraging Zebrafish To Study Bona Fide Astrocytes.
In a recent study, Chen and colleagues demonstrated that zebrafish spinal cord radial glia differentiate into cells that are similar to mammalian astrocytes. This study highlights the validity of the zebrafish model for discovering molecular mechanisms governing astrocyte function.


Immunohistochemical detection of carcinoembryonic antigen in benign hyperplasia and adenocarcinoma of the prostate with monoclonal antibody.
Immunoperoxidase localization of carcinoembryonic antigen was done on tissue sections of benign hyperplasia and adenocarcinoma of the prostate using murine monoclonal antibody against carcinoembryonic antigen. The study was done to determine whether carcinoembryonic antigen would be a tissue marker of prostatic malignancy. Of 30 benign lesions 25 (83 per cent) were negative and 5 had focally weak staining, while of 36 adenocarcinomas 31 (86 per cent) had diffusely positive staining and 5 were negative. Diffusely moderate or strong staining was noted in 5 of 10 grade I (50 per cent), 11 of 12 (92 per cent) grade II and 13 of 14 (93 per cent) grade III tumors, as well as 14 of 15 (93 per cent) tumors from patients with bone metastasis. These findings suggested that extensive immunostaining for carcinoembryonic antigen in prostatic glandular lesions might be a potentially important prognostic marker of malignancy. Further studies are warranted to extend and confirm these results.


Diagnostic ultrasound of the chest wall in the symptomatic patient after mastectomy.
Lack of standardized imaging recommendations among mastectomy patients has led to variability in how recurrences are detected. To describe the findings and assess the diagnostic efficacy of ultrasound in the evaluation of symptomatic post-mastectomy patients. This single institution, retrospective study included 749 consecutive diagnostic chest wall ultrasound examinations performed in mastectomy patients, from January 2016 to June 2017. Chest wall ultrasound evaluated the mastectomy bed with or without reconstruction. Electronic health records were queried for the primary breast cancer histology prior to mastectomy, clinical symptoms prompting the diagnostic ultrasound, ultrasound findings, subsequent cytology and pathology, and follow-up data. Excluded were patients with a known recurrence, asymptomatic patients, and those with <2 years of clinical or imaging follow-up. Descriptive and comparative statistical analyses were performed. Among the 749 ultrasounds performed, 58 malignancies were identified for a 7.7% (58/749) malignancy rate, with a median tumor size of 20 mm. Patients diagnosed with a malignancy most often presented with a palpable abnormality (79.3%, 46/58) or skin changes (13.8%, 8/58) and rarely with pain (1.7%, 1/58). Patients who underwent a biopsy yielding a benign result most often presented with a palpable abnormality (41.5%, 287/691), pain (25.6%,177/691), or postoperative swelling/suspected fluid collection (17.8%, 123/691). Diagnostic ultrasound yielded a 91.4% sensitivity (95% CI 81.0, 97.1), 96.1% specificity (95% CI 94.4, 97.4), 66.3% PPV3 (95% CI 57.4, 74.1), and 99.3% negative predictive value (95% CI 98.3, 99.7) for cancer detection. There were 5 false negative ultrasound cases after a skin punch biopsy was performed due to clinically suspicious skin changes. Chest wall ultrasound has a high sensitivity and negative predictive value for detection of breast cancer recurrence in symptomatic patients after mastectomy. Skin changes remain an important clinical manifestation of a cancer recurrence.


Allele frequency distribution of 1691G >A F5 (which confers Factor V Leiden) across Europe, including Slavic populations.
The allele 1691A F5, conferring Factor V Leiden, is a common risk factor in venous thromboembolism. The frequency distribution for this allele in Western Europe has been well documented; but here data from Central, Eastern and South-Eastern Europe has been included. In order to assess the significance of the collated data, a chi-squared test was applied, and Tukey tests and z-tests with Bonferroni correction were compared. A distribution with a North-Southeast band of high frequency of the 1691A F5 allele was discovered with a pocket including some Southern Slavic populations with low frequency. European countries/regions can be arbitrarily delimited into low (group 1, <2.8 %, mean 1.9 % 1691A F5 allele) or high (group 2, ≥2.8 %, mean 4.0 %) frequency groups, with many significant differences between groups, but only one intra-group difference (the Tukey test is suggested to be superior to the z-tests). In Europe a North-Southeast band of 1691A F5 high frequency has been found, clarified by inclusion of data from Central, Eastern and South-Eastern Europe, which surrounds a pocket of low frequency in the Balkans which could possibly be explained by Slavic migration. There seem to be no indications of variation in environmental selection due to geographical location.


The pharmacokinetics, tissue distribution, and biotransformation of a new class of antitumor agents: mitonafide and pinafide.
The pharmacokinetics, biotransformation, protein binding and tissue distribution of mitonafide and pinafide were studied after single i.v. and oral administration of each drug (20 mg/Kg) in female rats. In pregnant rats a study of cross placental-barrier after i.v. administration of the two drugs was also performed. The drugs were absorbed fairly rapidly with a mean peak plasma level of 7.63 +/- 0.70 micrograms eq/ml for the 3H-mitonafide and with 6.16 +/- 0.77 micrograms eq/ml for the 3H-pinafide between 30 minutes and 1 hour after oral dosing. For both drugs, the pharmacokinetics can be described by a two-compartment open model. The mean elimination half-lives were 17.8 h and 47.5 h for 3H-mitonafide and 3H-pinafide, respectively. Two metabolites for each compound as well as unchanged drugs were identified in urine by TLC and GC/MS by comparison of their chromatographic properties with a number of reference compounds. Approximately 30% of the radioactive drug was excreted over a 48 h period for 3H-mitonafide and 24% for 3H-pinafide in urine, after i.v. administration. The cross placental-barrier studies showed that both 3H-mitonafide and 3H-pinafide were present in the 14-day fetuses.


Clinical trials reimagined.
Clinical trials have been a central driver of change and have provided the evidence base necessary to advance new therapies for liver diseases. This review provides a perspective on the status of trials in hepatology and a vantage point into the emerging capabilities and external forces that will shape the conduct of clinical trials in the future. The adaptations to clinical trial operations in response to the disruptions by the COVID-19 pandemic and opportunities for innovation in hepatology trials are emphasized. Future trials in hepatology will be driven by unmet therapeutic needs and fueled by technological advances incorporating digital capabilities with expanded participant-derived data collection, computing, and analytics. Their design will embrace innovative trial designs adapted to these advances and that emphasize broader and more inclusive participant engagement. Their conduct will be further shaped by evolving regulatory needs and the emergence of new stakeholders in the clinical trials ecosystem. The evolution of clinical trials will offer unique opportunities to advance new therapeutics that will ultimately improve the lives of patients with liver diseases.


Post-transplantation nephrosis in congenital nephrotic syndrome of the Finnish type.
Congenital nephrotic syndrome of the Finnish type (CNF) is an autosomal recessively inherited disease manifesting as massive proteinuria, edema and ascites in the neonatal period. The disease is believed to be limited to the kidneys and recurrences after renal transplantation have not been reported. At our center 29 transplantations have been performed on 28 CNF patients. One to 33 months after transplantation, seven grafts (24%) of six patients have developed a steroid-resistant nephrotic syndrome. The clinical data and renal histology of these patients were analyzed in order to elucidate the cause of the proteinuria. At the onset of six of the seven episodes of nephrosis, the patient had evidence of a preceding CMV- or EBV-infection and the remaining patient had sinusitis. Upon light and electron microscopy examination, endothelial swelling of the glomerular capillaries resembling transplant glomerulopathy (TG) was seen, but unlike TG, the glomerular basement membranes were normal. The response of proteinuria to steroid or cyclophosphamide therapy was poor, with total remission in only two patients and partial remission in one patient, all treated with methylprednisolone and cyclophosphamide immediately after the diagnosis. Four grafts have been lost. Our data show that CNF patients have an increased tendency for post-transplantation nephrosis.


Effect on cerebral D-[3H]aspartate release of 3-mercaptopropionic acid and methyl-6,7-dimethoxy-4-ethyl-beta-carboline-3-carboxylate.
Minislices of rat brain cortex were preloaded with D-[3H]aspartate and spontaneous and potassium-induced (25 mM KCl) efflux of labelled compound into superfusate measured. 3-Mercaptopropionic acid, 100-220 microM did not modify the spontaneous efflux of D-[3H]aspartate, but augmented the K+-induced efflux. Methyl-6,7-dimethoxy-4-ethyl-beta-carboline-3-carboxylate, 10 or 100 microM, enhanced both spontaneous and K+-induced efflux of D-[3H]aspartate. The convulsant action of both compounds may be due to enhanced synaptic release of aspartate or a related excitatory neurotransmitter.


Proton transfer in the quinol-dependent nitric oxide reductase from Geobacillus stearothermophilus during reduction of oxygen.
Bacterial nitric oxide reductases (NOR) are integral membrane proteins that catalyse the reduction of nitric oxide to nitrous oxide, often as a step in the process of denitrification. Most functional data has been obtained with NORs that receive their electrons from a soluble cytochrome c in the periplasm and are hence termed cNOR. Very recently, the structure of a different type of NOR, the quinol-dependent (q)-NOR from the thermophilic bacterium Geobacillus stearothermophilus was solved to atomic resolution [Y. Matsumoto, T. Tosha, A.V. Pisliakov, T. Hino, H. Sugimoto, S. Nagano, Y. Sugita and Y. Shiro, Nat. Struct. Mol. Biol. 19 (2012) 238-246]. In this study, we have investigated the reaction between this qNOR and oxygen. Our results show that, like some cNORs, the G. stearothermophilus qNOR is capable of O(2) reduction with a turnover of ~3electronss(-1) at 40°C. Furthermore, using the so-called flow-flash technique, we show that the fully reduced (with three available electrons) qNOR reacts with oxygen in a reaction with a time constant of 1.8ms that oxidises the low-spin heme b. This reaction is coupled to proton uptake from solution and presumably forms a ferryl intermediate at the active site. The pH dependence of the reaction is markedly different from a corresponding reaction in cNOR from Paracoccus denitrificans, indicating that possibly the proton uptake mechanism and/or pathway differs between qNOR and cNOR. This study furthermore forms the basis for investigation of the proton transfer pathway in qNOR using both variants with putative proton transfer elements modified and measurements of the vectorial nature of the proton transfer. This article is part of a Special Issue entitled: 17th European Bioenergetics Conference (EBEC 2012).


Transcription factors TEAD2 and E2A globally repress acetyl-CoA synthesis to promote tumorigenesis.
Acetyl-coenzyme A (acetyl-CoA) plays an important role in metabolism, gene expression, signaling, and other cellular processes via transfer of its acetyl group to proteins and metabolites. However, the synthesis and usage of acetyl-CoA in disease states such as cancer are poorly characterized. Here, we investigated global acetyl-CoA synthesis and protein acetylation in a mouse model and patient samples of hepatocellular carcinoma (HCC). Unexpectedly, we found that acetyl-CoA levels are decreased in HCC due to transcriptional downregulation of all six acetyl-CoA biosynthesis pathways. This led to hypo-acetylation specifically of non-histone proteins, including many enzymes in metabolic pathways. Importantly, repression of acetyl-CoA synthesis promoted oncogenic dedifferentiation and proliferation. Mechanistically, acetyl-CoA synthesis was repressed by the transcription factors TEAD2 and E2A, previously unknown to control acetyl-CoA synthesis. Knockdown of TEAD2 and E2A restored acetyl-CoA levels and inhibited tumor growth. Our findings causally link transcriptional reprogramming of acetyl-CoA metabolism, dedifferentiation, and cancer.


Cycloartenyl Ferulate Inhibits Paraquat-Induced Apoptosis in HK-2 Cells With the Involvement of ABCC1.
Nephrotoxicity induced by chemicals such as paraquat (PQ) is a common clinical phenomenon; therefore, searching for drugs with renal protective effect is of a great practical significance. Our previous investigation found that cycloartenyl ferulate (CF) can antagonize the cytotoxic effect of PQ, and recent studies also revealed a variety of bioactivities of CF. However, specific molecular mechanisms underlying the protective effect of CF have not been explored yet. HPLC detection of PQ content indicated that CF reduced PQ accumulation in HK-2 cells and thereby improved cell survival. Western blot results showed that both PQ and CF did not affect the expression of ABCB1; however, while PQ suppressed the expression of ABCC1, CF upregulated ABCC1 expression and thereby reversed the inhibitory effect of PQ on ABCC1 expression. Meanwhile, HK-2 cells did not express ABCG2. When the expression of ABCC1 was knocked down with siRNA, the inhibitory effect of CF on intracellular PQ accumulation was blocked. Further flow cytometric analysis showed that while PQ significantly induced the appearance of sub-G1 apoptotic peak in cells, CF evidently inhibited apoptosis. TUNEL-DAPI double-staining also detected that PQ significantly induced the occurrence of DNA fragmentation in cells, whereas CF effectively inhibited the effect of PQ. Further results showed that ABCC1 siRNA effectively abolished the protective effect of CF on PQ-induced apoptosis. Taken together, these data demonstrated that in HK-2 cells, CF could antagonize PQ-induced toxicity with the involvement of regulatiion of ABCC1 protein expression, which provides a new strategy for treatments of nephrotoxicity.


A comprehensive review of quantum bioreactor cell manufacture: Research and clinical applications.
The Quantum cell expansion system manufactured by Terumo-BCT is perhaps the most widely reported Good Manufacturing Practice-compliant bioreactor used for the expansion of adherent cell populations, both for research purposes and clinical cell-based therapies/trials. Although the system was originally designed for adherent cell expansion, more recently suspension cultures and extracellular vesicle manufacturing protocols have been published using the Quantum system. Cell therapy research and regenerative medicine in general is a rapidly expanding field and as such it is likely that the use of this system will become even more widespread and perhaps mandatory, for both research and development and in the clinic. The purpose of this review is to describe, compare and discuss the diverse range of research and clinical applications currently using the Quantum system, which to our knowledge has not previously been reviewed. In addition, current and future challenges will also be discussed.


Prediction of fetal anemia by measurement of the mean blood velocity in the fetal aorta.
In 68 red blood cell isoimmunized pregnancies the mean velocity of blood in the fetal aorta was measured by pulsed Doppler ultrasonography, and the fetal hemoglobin concentration was determined in blood samples obtained by cordocentesis. The values were compared to reference ranges of mean aortic velocity and hemoglobin for gestation, which were determined from 218 218 pregnancies that were not complicated by fetal hemolysis. In the red blood cell isoimmunized pregnancies there was a significant positive correlation between the aortic mean velocity and the hemoglobin deficit for the nonhydropic fetuses and a significant negative correlation between these two parameters for the hydropic fetuses.