Discovery of new oxadiazolo pyridine derivatives as potent ghrelin O-acyltransferase inhibitors using molecular modeling techniques.
Diabesity is a major global health concern, and ghrelin O-acyltransferase (GOAT) acts as an important target for the development of new inhibitors of this disease. The present work highlights a detailed QSAR study using QSARINS software, which provides an excellent model equation using descriptors. Here, the best model equation developed has two variables, namely MLFER_E and XlogP, with statistical parameters R2 = 0.8433, LOF = 0.0793, CCCtr = 0.915, Q2LOO = 0.8303, Q2LMO = 0.8275, CCCcv = 0.9081, R2ext = 0.7712, and CCCext = 0.8668. A higher correlation of the key structural fragments with activity is validated by the developed QSAR model. Furthermore, molecular docking helped us identify the binding interactions. Thirty four new molecules with better predicted biological activity (pIC50) were designed. The binding energy of four compounds have shown higher binding activity into the membrane protein (PDB Id: 6BUG). Molecular dynamics simulation has established the stability of the protein-ligand complex over 100 ns. DFT and ADME-toxicity analyses also confirmed their drug-like properties. Based on our findings, we report that these new oxadiazolo pyridine derivatives lead to the development of potent candidates for further development. METTL3-mediated HOTAIRM1 promotes vasculogenic mimicry in glioma via regulating IGFBP2 expression. METTL3 expression is high in glioma cells and tissues that stabilize and enhance HOTAIRM1 expression. This HOTAIRM1 then interacts with IGFBP2 which in turn promotes glioma cell malignancy and vasculogenic mimicry (VM) formation, thus providing a new direction for glioma therapy. The online version contains supplementary material available at 10.1007/s40203-023-00167-z.


Cerebral perfusion disturbances in chronic mild traumatic brain injury correlate with psychoemotional outcomes.
The study explored associations between hemodynamic changes and psychoemotional status in 32 patients with chronic mild traumatic brain injury (mTBI) and 31 age-matched healthy volunteers. Cerebral blood flow (CBF) and cerebral blood volume (CBV) values were obtained using Dynamic Susceptibility Contrast Magnetic Resonance Imaging in brain regions suspected to play a role in anxiety and depression. Patients were administered self-report measures of anxiety and depression symptoms and underwent neuropsychological assessment. As a group mTBI patients scored significantly below age- and education-adjusted population norms on multiple cognitive domains and reported high rates of anxiety and depression symptomatology. Significantly reduced CBF values were detected in the mTBI group compared to controls in dorsolateral prefrontal areas, putamen, and hippocampus, bilaterally. Within the mTBI group, depressive symptomatology was significantly associated with lower perfusion in the left anterior cingulate gyrus and higher perfusion in the putamen, bilaterally. The latter association was independent from verbal working memory capacity. Moreover, anxiety symptomatology was associated with lower perfusion in the hippocampus (after controlling for verbal episodic memory difficulties). Associations between regional perfusion and psychoemotional scores were specific to depression or anxiety, respectively, and independent of the presence of visible lesions on conventional MRI. Results are discussed in relation to the role of specific limbic and paralimbic regions in the pathogenesis of symptoms of depression and anxiety.


Next-generation sequencing of tissue and circulating tumor DNA: Resistance mechanisms to EGFR targeted therapy in a cohort of patients with advanced non-small cell lung cancer.
Epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) has been considered as an effective treatment in epidermal growth factor receptor-mutant (EGFR-mutant) advanced non-small cell lung cancer (NSCLC). However, most patients develop acquired resistance eventually. Here, we compared and analyzed the genetic alterations between tissue assay and circulating tumor DNA (ctDNA) and further explored the resistance mechanisms after EGFR-TKI treatment. Amplification refractory mutation system-polymerase chain reaction (ARMS-PCR), Cobas® ARMS-PCR and next-generation sequencing (NGS) were performed on tissue samples after pathological diagnosis. Digital droplet PCR (ddPCR) and NGS were performed on plasma samples. The association between genetic alterations and clinical outcomes was analyzed retrospectively. Thirty-seven patients were included. The success rate of re-biopsy was 91.89% (34/37). The total detection rate of EGFR T790M was 62.16% (23/37) and the consistency between tissue and ctDNA was 78.26% (18/23). Thirty-four patients were analyzed retrospectively. For tissue re-biopsy, 24 patients harbored concomitant mutations. Moreover, tissue re-biopsy at resistance showed 21 patients (21/34, 61.76%) had the concomitant somatic mutation. The three most frequent concomitant mutations were TP53 (18/34, 52.94%), MET (4/34, 11.76%), and PIK3CA (4/34, 11.76%). Meanwhile, 21 patients (21/34, 61.76%) with EGFR T790M mutation. Progression-free survival (PFS) and overall survival (OS) were better in patients with T790M mutation (p = 0.010 and p = 0.017) or third-generation EGFR-TKI treatment (p < 0.0001 and p = 0.073). Interestingly, concomitant genetic alterations were significantly associated with a worse prognosis for patients with EGFR T790M mutation receiving third-generation EGFR-TKIs (p = 0.037). Multi-platforms are feasible and highly consistent for re-biopsy after EGFR-TKI resistance. Concomitant genetic alterations may be associated with a poor prognosis for patients with EGFR T790M mutation after third-generation EGFR-TKIs.


Mediatization of development in sub Saharan Africa: insights from Ethiopian Orthodox Tewahedo Church (EOTC), 'Mahibere Kidusan' magazine, Ethiopia.
Development is an important issue for third world Sub-Saharan African countries such as Ethiopia. To enhance development, media have a significant role, but faith-based institutions, like religious media have not been considered significant contributors so far. This paper studies the Hamer, a religious magazine in Ethiopia Orthodox Tewahedo (EOTC) for about 30 years. This research examines the agenda and frames used by the Hamer magazine coverage of development issues (from 2005 -2020) in Ethiopia. The study applies a quantitative content analysis method and examined 308 stories. However, the political development issue was the most scarcely covered development issue as compared to others. Economic, moral, peace and social development issues had a relationship with the year of publication. Sources of economic development stories and dominant frames had impact on the choice of economic development stories. Similarly, dominant frames had impact on the choice of peace development stories. On the contrary, sources, story actors and dominant frames did not have significant impact on social, political, cultural, historical, moral, and natural resource development stories. Finally, the Hamer magazine had significant coverage of development issues although it is difficult to determine the causes of the extent of changes in the coverage. In conclusion, we propose that governmental and nongovernmental organizations should reconsider their communication contexts and can reach disadvantages through religious media while launching development programs.


Vascular trauma: successful repair using human umbilical cord vein interposition grafts in femoral artery and vein.
A case of extensive injury to the left femoral artery and vein is presented. As circumstances did not permit the use of autogenous vein grafts, human umbilical cord vein grafts were used. Almost one year afterwards the patient is doing well and both grafts are patent.


Presence of an insulin-stimulated serine kinase in cell extracts from IM-9 cells.
Insulin responsive protein kinase activities of wheat germ purified glycoproteins were examined. Glycoproteins were first incubated without or with insulin, and then exposed to a serum containing antibodies to insulin receptor. Thereafter, both immunoprecipitates and supernatants were studied for their kinase activity toward histone. Incubation with anti receptor antibodies promoted insulin receptor beta subunit and histone phosphorylation. More important insulin receptor depleted extract contained a kinase activity toward histone, that was increased by preincubation with insulin. This stimulation was observed only when insulin was added before the immunoprecipitation of insulin receptors. Alkali treatment and phosphoamino acids analysis revealed that the kinase activity remaining in the supernatant is serine specific. These findings suggest, that a serine kinase activity is associated with the insulin receptor, that it can be separated from the insulin receptor with anti receptor antibodies, that the serine kinase is activated by the hormone-receptor complex.


Increased incidence of stings in venom-sensitive patients.
We compared the histories of 29 venom-sensitive and 28 control subjects who were selected from our venom referral and general allergy clinics respectively. The variables in the study included insect avoidance knowledge, the number of stings during the previous 2 years, insects involved, and time spent out of doors per week. There was no significant difference between the two groups with respect to age. All venom-sensitive patients were well versed in avoidance techniques while only 3 of 28 controls (11%) claimed such knowledge. Venom-sensitive subjects were stung almost ten times more frequently than control subjects. Wasp stings were the most common, followed by yellow jacket, honey bee, and hornet. The venom-sensitive patients also reported spending a greater amount of time outdoors (x 17.4 hours versus x 11.8, P < .05). An analysis of covariance showed that this difference in outdoor exposure was insufficient to account for the disparity in the number of stings. We conclude that other factors such as intrinsic attractants must be responsible for this phenomenon.


Position of shoulder arthroplasty and clinical outcome in proximal humerus fractures.
Four-part proximal humeral fractures are frequently treated with shoulder replacement. Outcome of this procedure has not been standardized, and there are controversy data on range of motion (ROM) and active function of the shoulder. The aim of this study is to compare shoulder prosthesis position (SPP) in terms of version of humeral head and height of stem with clinical subjective and objective outcome. Fifty patients were treated with shoulder hemiarthroplasty for four-part proximal humeral fracture or fracture-dislocation of the humeral head. Radiological examination and CT-scan were performed preoperatively and at follow-up. Clinical outcome evaluation included active and passive ROM, and subjective perspective collected through SF-36, OSQ, ASES, and DASH. No significant correlation between stem height and clinical outcome were found. The prosthesis version correlates with all subjective questionnaires. The ROM was not correlated with stem height and prosthesis version. SPP involves clinical outcome, with great relevance of implant version.


Determining Gut Microbial Dysbiosis: a Review of Applied Indexes for Assessment of Intestinal Microbiota Imbalances.
Assessing "dysbiosis" in intestinal microbial communities is increasingly considered a routine analysis in microbiota studies, and it has added relevant information to the prediction and characterization of diseases and other adverse conditions. However, dysbiosis is not a well-defined condition. A variety of different dysbiosis indexes have been suggested and applied, but their underlying methodologies, as well as the cohorts and conditions for which they have been developed, differ considerably. To date, no comprehensive overview and comparison of all the different methodologies and applications of such indexes is available. Here, we list all types of dysbiosis indexes identified in the literature, introduce their methodology, group them into categories, and discuss their potential descriptive and clinical applications as well as their limitations. Thus, our focus is not on the implications of dysbiosis for disease but on the methodological approaches available to determine and quantify this condition.


Corrigendum: Methylation of FKBP5 and SLC6A4 in Relation to Treatment Response to Mindfulness Based Stress Reduction for Posttraumatic Stress Disorder.
[This corrects the article DOI: 10.3389/fpsyt.2018.00418.].


The discrepancy between plasma clearance tests of indocyanine green (ICG) and sulfobromophthalein (BSP): --report of cases and a study of ICG-binding pattern of serum proteins.
Two cases of Gilbert's syndrome and three cases of other diseases showed abnormally low plasma indocyanine green (ICG) clearance as contrated with normal or nearly normal plasma sulfobromphthalein (BSP) clearance. To evaluate the role of serum proteins in pathogenesis of these cases, ICG-binding patterns of serum proteins of the patients were compared with that of normal persons by the method of Sephadex G200 column chromatography. No qualitative differences in binding patterns were observed between serum proteins of the patients and normal persons. From these results, serum proteins were thought to be minor determinants of the hepatic ICG uptake mechanism. Because of normal or nearly normal plasma BSP clearance in these patients, the contents of hepatic organic anion-binding proteins, Y and Z, were thought to be sufficient. Possible qualitative abnormalities of the Y and/or Z proteins were also discussed.


A methodology for incorporating functional bone marrow sparing in IMRT planning for pelvic radiation therapy.
The purpose of this study was to design a radiation therapy treatment planning approach that would spare hematopoietically active bone marrow using [(18)F]FLT PET imaging. We have developed an IMRT planning methodology to incorporate functional PET imaging using [(18)F]FLT scans. Plans were generated for two simulated cervical cancer patients, where pelvic active bone marrow regions were incorporated as avoidance regions based on the ranges: SUV4 ≥ 4; 4>SUV3 ≥ 3; and 3 > SUV2 ≥ 2. Dose objectives were set to reduce bone marrow volume that received 10 (V(10)) and 20 (V(20))Gy. Active bone marrow regions identified by [(18)F]FLT with an SUV ≥ 2, SUV ≥ 3, and SUV ≥ 4 represented an average of 43.0%, 15.3%, and 5.8%, respectively of the total osseous pelvis for the two cases studied. Improved dose-volume histograms for all identified bone marrow SUV volumes and decreases in V(10), and V(20) were achieved without clinically significant changes to PTV or OAR doses. Incorporation of [(18)F]FLT PET in IMRT planning provides a methodology to reduce radiation dose to active bone marrow without compromising PTV or OAR dose objectives in pelvic malignancies.


Impact of Mechanical Complications on Success of Dental Implant Treatments: A Case-Control Study.
This study aimed to investigate the impact of mechanical complications on outcome measures for implant dentistry. This case-control study included 282 patients with mechanical complications occurring in fixed prosthetic rehabilitation supported by immediate function implants with external connection (cases) and 282 individuals without mechanical complications (control). Pairing was performed for sex, age (range = 3 years), and follow-up months (range = 11 months). The primary outcome measure was implant survival, while the secondary outcome measures were marginal bone loss and biological complication parameters (peri-implant pathology, soft tissue inflammation, fistula formation, and abscess formation). Cumulative implant survival was estimated by using life tables. Descriptive statistics with 95% confidence intervals (CI) and inferential statistics (Chi-square test) were performed to evaluate differences between cases and controls. The significance level was set at 5%. The average follow-up duration was 8.5 years. Mechanical complications included prosthetic fracture (n = 159), abutment loosening (n = 89), prosthetic screw loosening (n = 20), milled abutment (n = 12), milled prosthetic screw (n = 1), and decemented crown (n = 1). Implant failure occurred in one patient from the control group, with survival rates of 100 and 99.6% for cases and controls, respectively (p = 0.317). The average marginal bone loss was 1.72 (95% confidence interval [CI]: 1.60-1.84) for cases and 1.55 (95% CI: 1.45-1.65) for controls (p = 0.068). Biological complications were observed in 90 patients, with significant differences between cases (n = 54) and controls (n = 36; p = 0.038). Mechanical complications did not significantly influence survival or marginal bone loss; nevertheless, there is a need for studies with longer follow-up duration. Mechanical complications also significantly influence the incidence of biological complications.


Characterization of protein components of human urinary crystal surface binding substance.
We previously extracted crystal surface binding substance (CSBS) from human urine and showed that it appeared to constitute a substantial proportion of urinary macromolecular inhibitors of calcium oxalate crystallization. CSBS was isolated from human urine and fractionated by three consecutive chromatography procedures in order to characterize protein inhibitors of calcium oxalate crystallization. Sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and NH2-terminal amino acid sequencing revealed that inhibitory fractions eluted from a final, hydroxyapatite column contained prothrombin and osteopontin. Hydroxyapatite column fractions also contained other, unidentified protein inhibitors of calcium oxalate crystallization. CSBS contained also human serum albumin, alpha 1-acid glycoprotein, alpha 1-microglobulin, alpha 2-HS glycoprotein, retinol-binding protein, transferrin, and Tamm-Horsfall protein, but these proteins seemed to play no direct role in inhibitory activity.


HLA-HD: An accurate HLA typing algorithm for next-generation sequencing data.
The accurate typing of human leukocyte antigen (HLA) alleles is critical for a variety of medical applications, such as genomic studies of multifactorial diseases, including immune system and inflammation-related disorders, and donor selection in organ transplantation and regenerative medicine. Here, we developed a new algorithm for determining HLA alleles using next-generation sequencing (NGS) results. The method consists of constructing an extensive dictionary of HLA alleles, precise mapping of the NGS reads, and calculating a score based on weighted read counts to select the most suitable pair of alleles. The developed algorithm compares the score of all allele pairs, taking into account variation not only in the domain for antigen presentation (G-DOMAIN), but also outside this domain. Using this method, HLA alleles could be determined with 6-digit precision. We showed that our method was more accurate than other NGS-based methods and revealed limitations of the conventional HLA typing technologies. Furthermore, we determined the complete genomic sequence of an HLA-A-like-pseudogene when we assembled NGS reads that had caused arguable typing, and found its identity with HLA-Y*02:01. The accuracy of the HLA-A allele typing was improved after the HLA-Y*02:01 sequence was included in the HLA allele dictionary.


Endoscopic and Surgical Treatments for Gastroparesis: What to Do and Whom to Treat?
Gastroparesis is a complex chronic debilitating condition of gastric motility resulting in the delayed gastric emptying and multiple severe symptoms, which may lead to malnutrition and dehydration. Initial management of patients with gastroparesis focuses on the diet, lifestyle modification and medical therapy. Various endoscopic and surgical interventions are reserved for refractory cases of gastroparesis, not responding to conservative therapy. Pyloric interventions, enteral access tubes, gastric electrical stimulator and gastrectomy have been described in the care of patients with gastroparesis. In this article, the authors review current management, indications, and contraindications to these procedures.


When day meets night: Subsiding calcium signalling translates daylight into new neurones.
Day/night cycle controls neurogenesis; melatonin released from pineal gland in darkness stimulates intracellular Ca2+ dynamics thus decreasing proliferation of neural stem cells. In the daylight intracellular Ca2+ activity subsides, which stimulates neural stem cells division and increases generation of newborn neurones.


Sidon on the breadth of the wild sea: Movement and diet on the Mediterranean coast in the Middle Bronze Age.
Excavations at Sidon (Lebanon) have revealed dual identities during the Middle Bronze Age (ca. 2000-1600 BCE): a maritime port and center for local distribution, as well as a settlement with a heavy subsistence dependence on the extensive inland hinterlands. We aim to investigate residential mobility at Sidon using isotopic analyses of 112 individuals from 83 burials (20 females, 26 males, and 37 subadults). Veneration and remembrance of the dead is evident from funerary offerings in and near the tombs. With marine fish a major component in funerary offerings, we predict major marine reliance in this coastal population. New isotopic evidence of paleomobility (87 Sr/86 Sr, δ18 O) and diet (δ13 Ccarbonate ) is the focus of this research. Previous bulk bone collagen δ13 C and δ15 N analysis is strengthened by further sampling, along with δ34 S where collagen yield was sufficient. The five non-locals identified (8.9% of the 56 analyzed) come from constructed tombs with high-status grave goods except for one, which was heavily disturbed in antiquity. Dietary investigation of the population confirms reliance on terrestrial resources with no significant marine input. No significant differences in diet between the sexes or burial types are present. Although Sidon was part of a growing Mediterranean network evidenced through artefactual finds, relatively low immigration is evident. While religious feasts venerating the dead may have involved significant piscine components, no appreciable marine input in diet is observed. Fish may have been reserved for the deceased or only consumed on feast days alongside the dead rather than a regular part of the Bronze Age menu.


Relationship between oral health-related knowledge, attitudes and behavior among 15-16-year-old adolescents: a structural equation modeling approach.
The aim of this study was to confirm the previously observed attitudinal factor structure related to behavioral change and the knowledge-attitude-behavior model on dental health and hygiene among adolescents. The study population consisted of all 8(th) and 9(th) graders (15-16 years) who started the 2004-2005 school year in Rauma, Finland (n = 827). Data on knowledge, attitudes, toothbrushing and using fluoride toothpaste were gathered by questionnaires. Hypothesized structure included four attitudinal factors related to dental health and hygiene: 'importance of toothbrushing when participating in social situations' (F1), 'importance of toothbrushing for health-related reasons and better appearance' (F2), 'being concerned about developing caries lesions' (F3) and 'importance of toothbrushing for feeling accepted' (F4). Structural equation modeling (SEM) was used to test the hypothesized model: pathways lead from knowledge to behavior both directly and via attitudes. The hypothesized model was also modified by removing non-significant pathways and studying the inter-relationships between attitudes. A confirmatory factor analysis revealed that factor F4 had to be removed. In the final model, knowledge influenced behavior directly and via two attitude factors, F1 and F2, which were inter-related. 'Concern about developing caries lesions' was a background factor influencing only knowledge. The final factor structure and SEM model were acceptable-to-good fit. Knowledge had a smaller effect on behavior than on attitudes. Our results support theories about the causal knowledge-attitudes-behavior chain, also for adolescents' oral health-related behaviors.


Conflict and surrender during sentence processing: an ERP study of syntax-semantics interaction.
Recent ERP studies report that implausible verb-argument combinations can elicit a centro-parietal P600 effect (e.g., "The hearty meal was devouring …"; Kim & Osterhout, 2005). Such eliciting conditions do not involve outright syntactic anomaly, deviating from previous reports of P600. Kim and Osterhout (2005) attributed such P600 effects to structural reprocessing that occurs when syntactic cues fail to support a semantically attractive interpretation ('meal' as the Agent of 'devouring') and the syntactic cues are overwhelmed; the sentence is therefore perceived as syntactically ill-formed. The current study replicated such findings and also found that altering the syntactic cues in such situations of syntax-semantics conflict (e.g., "The hearty meal would devour …") affects the conflict's outcome. P600s were eliminated when sentences contained syntactic cues that required multiple morphosyntactic steps to "repair". These sentences elicited a broad, left-anterior negativity at 300-600ms (LAN). We interpret the reduction in P600 amplitude in terms of "resistance" of syntactic cues to reprocessing. We speculate that the LAN may be generated by difficulty retrieving an analysis that satisfies both syntactic and semantic cues, which results when syntactic cues are strong enough to resist opposing semantic cues. This pattern of effects is consistent with partially independent but highly interactive syntactic and semantic processing streams, which often operate collaboratively but can compete for influence over interpretation.


Distribution of repetitive DNA sequences in chromosomes of five opisthorchid species (Trematoda, Opisthorchiidae).
Genomes of opisthorchid species are characterized by small size, suggesting a reduced amount of repetitive DNA in their genomes. Distribution of repetitive DNA sequences in the chromosomes of five species of the family Opisthorchiidae (Opisthorchis felineus 2n = 14 (Rivolta, 1884), Opisthorchis viverrini 2n = 12 (Poirier, 1886), Metorchis xanthosomus 2n = 14 (Creplin, 1846), Metorchis bilis 2n = 14 (Braun, 1890), Clonorchis sinensis 2n = 14 (Cobbold, 1875)) was studied with C- and AgNOR-banding, generation of microdissected DNA probes from individual chromosomes and fluorescent in situ hybridization on mitotic and meiotic chromosomes. Small-sized C-bands were discovered in pericentric regions of chromosomes. Ag-NOR staining of opisthorchid chromosomes and FISH with ribosomal DNA probe showed that karyotypes of all studied species were characterized by the only nucleolus organizer region in one of small chromosomes. The generation of DNA probes from chromosomes 1 and 2 of O. felineus and M. xanthosomus was performed with chromosome microdissection followed by DOP-PCR. FISH of obtained microdissected DNA probes on chromosomes of these species revealed chromosome specific DNA repeats in pericentric C-bands. It was also shown that microdissected DNA probes generated from chromosomes could be used as the Whole Chromosome Painting Probes without suppression of repetitive DNA hybridization. Chromosome painting using microdissected chromosome specific DNA probes showed the overall repeat distribution in opisthorchid chromosomes.


Genetics and neurobiology of frontotemporal lobar degeneration.
Frontotemporal dementia (FTD) is characterised by brain intracellular deposition of abnormally phosphorylated tau protein, considered responsible for neuronal death. Several familial cases with different mutations in the tau encoding gene (MAPT), located on chromosome 17, have been described. Besides, in a Danish family, the genetic defect has been associated to chromosome 3. Although many FTD families exhibit known mutations, in some cases none of them occur. Recent findings demonstrate an increased intrathecal production of both pro- and anti-inflammatory cytokines in sporadic FTD patients. Besides, increased cerebrospinal fluid monocyte chemotactic protein-1 and interleukin-8 levels have been observed in FTD, whereas interferon-gamma-inducible protein-10 levels were similar to controls.


Resistance-Conferring Mutations on Whole-Genome Sequencing of Fluoroquinolone-resistant and -Susceptible Mycobacterium tuberculosis Isolates: A Proposed Threshold for Identifying Resistance.
Fluoroquinolone resistance in Mycobacterium tuberculosis (Mtb) is conferred by DNA gyrase mutations, but not all fluoroquinolone-resistant Mtb isolates have mutations detected. The optimal allele frequency threshold to identify resistance-conferring mutations by whole-genome sequencing is unknown. Phenotypically ofloxacin-resistant and lineage-matched ofloxacin-susceptible Mtb isolates underwent whole-genome sequencing at an average coverage depth of 868 reads. Polymorphisms within the quinolone-resistance-determining region (QRDR) of gyrA and gyrB were identified. The allele frequency threshold using the Genome Analysis Toolkit pipeline was ~8%; allele-level data identified the predominant variant allele frequency and mutational burden (ie, sum of all variant allele frequencies in the QRDR) in gyrA, gyrB, and gyrA + gyrB for each isolate. Receiver operating characteristic (ROC) curves assessed the optimal measure of allele frequency and potential thresholds for identifying phenotypically resistant isolates. Of 42 ofloxacin-resistant Mtb isolates, area under the ROC curve (AUC) was highest for predominant variant allele frequency, so that measure was used to evaluate optimal mutation detection thresholds. AUCs for 8%, 2.5%, and 0.8% thresholds were 0.8452, 0.9286, and 0.9069, respectively. Sensitivity and specificity were 69% and 100% for 8%, 86% and 100% for 2.5%, 91% and 91% for 0.8%. The sensitivity of the 2.5% and 0.8% thresholds were significantly higher than the 8% threshold (P = .016 and .004, respectively) but not significantly different between one another (P = .5). A predominant mutation allele frequency threshold of 2.5% had the highest AUC for detecting DNA gyrase mutations that confer ofloxacin resistance, and was therefore the optimal threshold.


Graft kidney hydronephrosis caused by transplant ureter inguinal hernia: A case report.
Ureteral obstruction of the graft kidney is a common complication of kidney transplantation. However, ureteral obstruction caused by inguinal hernia has rarely been reported. We present a rare case of ureteral obstruction with allograft dysfunction caused by an inguinal hernia. A 76-year-old man, who was a renal transplant recipient, presented with bilateral pitting oedema, reduced urine output, and right inguinal hernia. Abdominal computed tomography revealed severe hydroureteronephrosis of the kidney allograft. A right inguinal hernia with ureteral incarceration was observed. The patient underwent graft percutaneous nephrostomy, followed by antegrade insertion of a double-J tube (DJ). Gradual improvement was observed in his renal function. Right inguinal herniorrhaphy was performed 5 days later. The renal function returned to normal after percutaneous nephrostomy and insertion of the DJ. A right inguinal direct-type hernia with ureter adhesion to the hernial sac was observed during the surgery. The posterior wall defect was repaired by the McVay technique. The DJ was removed after 1 month. The patient's renal function remained stable at 6-month follow-up. The orientation of the graft kidney has a significant influence on the location of the ureter. Upward orientation of the hilum will result in superficial location of the ureter, rendering it close to the hernial sac and susceptible to incarceration. The transplant surgeon should be aware of such a presentation of graft dysfunction with inguinal hernia to prevent a delay in the diagnosis and graft loss.


Evaluation of Rhesus Macaque Models for Cerebral Palsy.
Animal models play a central role in all areas of biomedical research. The similarities in anatomical structure and physiological characteristics shared by non-human primates (NHPs) and humans make NHPs ideal models with which to study human disorders, such as cerebral palsy (CP). However, the methodologies for systematically evaluating NHP models of CP have rarely been assessed, despite the long history of using NHP models to understand CP. Such models should be evaluated using multidisciplinary approaches prior to being used to research the diagnosis and treatment of CP. In this study, we evaluated rhesus macaque CP models established by partial resection of the motor cortex and intrathecal injection of bilirubin. Abnormal posture, motor dysfunction, gross and fine motor behavior, and muscular tension were evaluated, and changes in the cerebral cortex and basal ganglia were observed using 9.4 T magnetic resonance imaging. The results clearly demonstrated the utility of the established evaluation methodology for assessing CP models. This model evaluation methodology may guide researchers through the model building process.


Innovation Is Tied to Optimism.
"At the height of laughter, the universe is flung into a kaleidoscope of new possibilities."-Jean Houston.


Zika Virus Targets Multiple Tissues and Cell Types During the First Trimester of Pregnancy.
The 2016 Zika virus (ZIKV) outbreak in the Americas has been characterized by an increased association frequency of fetal neuropathological abnormalities. To have a comprehensive and accurate knowledge of key elements of the clinically observed neurologic dysfunctions in Zika-infected babies, ZIKV transmission from mother to fetus needs to be deeply studied. Thus, it is important to determine the role of both virus-targeted cells and tissues within the mother-fetus interface. Cellular tropism and mechanisms of ZIKV transmission from mother to the fetus during early pregnancy still remain unknown on many aspects. To improve the characterization of the maternal-fetal ZIKV transmission, we have set up an ex vivo model using an organ culture approach with a light-invasive sampling from the first trimester of pregnancy samples. Thus, here we provide evidence that circulating epidemic ZIKV strains from Latin America widely target and destroy reproductive tissues, including the decidua basalis, fetal placenta, and umbilical cord. In addition, we show that ZIKV is able to differentially replicate in a large range of both maternal and fetal cells, including decidual fibroblasts and macrophages, fetal trophoblast and Hofbauer cells, as well as umbilical cord mesenchymal stem cells. This primary and broad ZIKV cellular tropism and the resulting abundant cytopathic-induced tissue effects during the first trimester of pregnancy show the upstream path of clinically observed congenital damages.


Inhibition of Vesicular Glutamate Transporters (VGLUTs) with Chicago Sky Blue 6B Before Focal Cerebral Ischemia Offers Neuroprotection.
Brain ischemia is one of the leading causes of death and long-term disability in the world. Interruption of the blood supply to the brain is a direct stimulus for many pathological events. The massive vesicular release of glutamate (Glu) after ischemia onset induces excitotoxicity, which is a potent stress on neurons. Loading of presynaptic vesicles with Glu is the first step of glutamatergic neurotransmission. Vesicular glutamate transporters 1, 2, and 3 (VGLUT1, 2, and 3) are the main players involved in filling presynaptic vesicles with Glu. VGLUT1 and VGLUT2 are expressed mainly in glutamatergic neurons. Therefore, the possibility of pharmacological modulation to prevent ischemia-related brain damage is attractive. In this study, we aimed to determine the effect of focal cerebral ischemia on the spatiotemporal expression of VGLUT1 and VGLUT2 in rats. Next, we investigated the influence of VGLUT inhibition with Chicago Sky Blue 6B (CSB6B) on Glu release and stroke outcome. The effect of CSB6B pretreatment on infarct volume and neurological deficit was compared with a reference model of ischemic preconditioning. The results of this study indicate that ischemia upregulated the expression of VGLUT1 in the cerebral cortex and in the dorsal striatum 3 days after ischemia onset. The expression of VGLUT2 was elevated in the dorsal striatum and in the cerebral cortex 24 h and 3 days after ischemia, respectively. Microdialysis revealed that pretreatment with CSB6B significantly reduced the extracellular Glu concentration. Altogether, this study shows that inhibition of VGLUTs might be a promising therapeutic strategy for the future.


A computational modeling approach for enhancing self-assembly and biofunctionalisation of collagen biomimetic peptides.
Collagen fibers are essential components of tissues, which are highly conserved across the animal kingdom and could be extremely useful in tissue engineering. The formation of these macromolecular fibers depends on molecular interactions-based self-assembly of the basic building blocks of collagen called tropocollagens. Several attempts to produce biomimetic collagen have been described, however the best method to achieve the optimal material for tissue engineering has not been established. Here, we describe a bottom-up approach to design two computationally mutated molecular models that use non-covalent interactions to cross-link triple helices of tropocollagen molecules and thus promote self-association. Implementing a graph theory approach in the software FIRST reveals the hotspots that are crucial for the overall rigidity of the supramolecular helical structures and the remaining non-hotspots available for mutations. The mutated models were further decorated with GFOGER, a known collagen cell binding motif, to depict a biofunctional model. In addition to their recognized role of cell binding, the charged residues of the binding motif appeared to enhance further the supramolecular helical association. These findings could help to produce biomimetic collagen for biomedical applications.


Vector competence of Anopheles stephensi for O'nyong-nyong virus: a risk for global virus spread.
O'nyong-nyong virus (ONNV) is a mosquito-borne alphavirus causing sporadic outbreaks of febrile illness with rash and polyarthralgia. Up to now, ONNV has been restricted to Africa and only two competent vectors have been found, Anopheles gambiae and An. funestus, which are also known malaria vectors. With globalization and invasive mosquito species migrating to ONNV endemic areas, there is a possible risk of introduction of the virus to other countries and continents. Anopheles stephensi, is closely related to An. gambiae and one of the invasive mosquito species of Asian origin that is now present in the Horn of Africa and spreading further east. We hypothesize that An. stephensi, a known primary urban malaria vector, may also serve as a new possible vector for ONNV. One-week-old female adult An. stephensi were exposed to ONNV-infected blood, and the vector competence for ONNV, i.e. infection rates (IRs), dissemination rates (DRs), transmission rates (TRs), dissemination efficiency (DEs) and transmission efficiency (TEs), were evaluated. Infection (IRs), dissemination efficiency (DEs) and transmission efficiency (TEs) were determined. Detection of ONNV RNA was analysed by RT-qPCR in the thorax and abdomen, head, wings, legs and saliva of the infected mosquitoes at four different time points, day 7, 14, 21 and 28 after blood meal. Infectious virus in saliva was assessed by infection of Vero B4 cells. The mean mortality across all sampling times was 27.3% (95 confidence interval [CI] 14.7-44.2%). The mean rate of infection across all sampling periods was 89.5% (95% CI 70.6-95.9). The mean dissemination rate across sampling intervals was 43.4% (95% CI 24.3-64.2%). The mean TR and TE across all mosquito sampling time intervals were 65.3 (95% CI 28.6-93.5) and 74.6 (95% CI 52.1-89.4). The IR was 100%, 79.3%, 78.6% and 100% respectively at 7, 14, 21 and 28 dpi. The DR was the highest at 7 dpi with 76.0%, followed by 28 dpi at 57.1%, 21 dpi at 27.3% and 14 dpi at the lowest DR of 13.04%. DE was 76%, 13.8%, 25%, 57.1% and TR was 79%, 50%, 57.1% and 75% at 7, 14, 21 and 28 dpi respectively. The TE was the highest at 28 dpi, with a proportion of 85.7%. For 7, 14 and 21 dpi the transmission efficiency was 72.0%, 65.5% and 75.0% respectively. Anopheles stephensi is a competent vector for ONNV and being an invasive species spreading to different parts of the world will likely spread the virus to other regions.


High-density mapping for gray leaf spot resistance using two related tropical maize recombinant inbred line populations.
Gray leaf spot (GLS) caused by Cercospora zeae-maydis or Cercospora zeina is one of the devastating maize foliar diseases worldwide. Identification of GLS-resistant quantitative trait loci (QTL)/genes plays an urgent role in improving GLS resistance in maize breeding practice. Two groups of recombinant inbred line (RIL) populations derived from CML373 × Ye107 and Chang7-2 × Ye107 were generated and subjected to genotyping-by-sequencing (GBS). A total of 1,929,222,287 reads in CML373 × Ye107 (RIL-YCML) and 2,585,728,312 reads in Chang7-2 × Ye107 (RIL-YChang), with an average of 10,961,490 (RIL-YCML) and 13,609,096 (RIL-YChang) reads per individual, were got, which was roughly equal to 0.70-fold and 0.87-fold coverage of the maize B73 RefGen_V4 genome for each F7 individual, respectively. 6418 and 5139 SNP markers were extracted to construct two high-density genetic maps. Comparative analysis using these physically mapped marker loci demonstrated a satisfactory colinear relationship with the reference genome. 11 GLS-resistant QTL have been detected. The individual QTL accounted for 1.53-24.00% of the phenotypic variance explained (PVE). The new consensus QTL (qYCM-DS3-3/qYCM-LT3-1/qYCM-LT3-2) with the largest effect was located in chromosome bin 3.05, with an interval of 2.7 Mb, representing 13.08 to 24.00% of the PVE. Further gene annotation indicated that there were four candidate genes (GRMZM2G032384, GRMZM2G041415, GRMZM2G041544, and GRMZM2G035992) for qYCM-LT3-1, which may be related to GLS resistance. Combining RIL populations and GBS-based high-density genetic maps, a new larger effect QTL was delimited to a narrow genomic interval, which will provide a new resistance source for maize breeding programs.


Prevalence of Dementia and Main Subtypes in Mexico: The Study on Aging and Dementia in Mexico (SADEM).
Dementia is a priority public health issue due to its high prevalence worldwide and its economic, social, and health impact. However, there are few reports in Mexico based on formal tests and with a clinical approach based on the Diagnostic and Statistical Manual of Mental Disorders, 5th edition (DSM-5). This study estimates the prevalence of the main types of dementia among elderly people living in the community in Mexico City. A population-based, two-step study was conducted, including 6,204 elderly individuals aged 60 or above with in-home assessment. All participants were screened for cognitive impairment; those who presented some cognitive problem underwent a standardized neurological examination. Each diagnosis was based on the criteria for dementia in the DSM-5, and the final consensus diagnosis of dementia was determined by an expert panel. The global estimated prevalence of dementia in the Mexican population was 7.8% met the criteria for Alzheimer's disease, 4.3% for vascular dementia, and 2.1% for mixed dementia. The prevalence of dementia was higher in women than in men (15.3% versus 12.5%, respectively). These results provide evidence to propose strategies for Latin American countries where dementia represents a challenge due to the heterogeneity of the populations and socioeconomic disparities, requiring early diagnosis and at the first levels of care.


A Generic Model for Follicular Lymphoma: Predicting Cost, Life Expectancy, and Quality-Adjusted-Life-Year Using UK Population-Based Observational Data.
To use real-world data to develop a flexible generic decision model to predict cost, life expectancy, and quality-adjusted life-years (QALYs) for follicular lymphoma (FL) in the general patient population. All patients newly diagnosed with FL in the UK's population-based Haematological Malignancy Research Network (www.hmrn.org) between 2004 and 2011 were followed until 2015 (N = 740). Treatment pathways, QALYs, and costs were incorporated into a discrete event simulation to reflect patient heterogeneity, including age and disease management. Two scenario analyses, based on the latest National Institute for Health and Clinical Excellence (NICE) guidelines (rituximab induction therapy for newly diagnosed asymptomatic patients and rituximab maintenance therapy for patients between treatments), were conducted and their economic impacts were compared to current practice. Incidence-based analysis revealed expected average lifetime costs ranging from £6,165 [US$7,709] to £63,864 [US$79,862] per patient, and average life expectancy from 75 days to 17.56 years. Prevalence-based analysis estimated average annual treatment costs of £60-65 million [US$75-80 million], accounting for approximately 10% of the United Kingdom's annual National Health Service budget for hematological cancers as a whole. Assuming that treatment effects reported in trials are applicable to all patient groups, scenario analyses for two recent NICE guidelines demonstrated potential annual cost savings for the United Kingdom that ranged with uptake frequency from £0.6 million to £11 million [US$0.75-2.75 million]. Costs, survival, and QALYs associated with FL vary markedly with patient characteristics and disease management. Allowing the production of more realistic outcomes across the patient population as a whole, our model addresses this heterogeneity and is a useful tool with which to evaluate new technologies/treatments to support healthcare decision makers.


Electrochemical properties of protoporphyrin IX zinc(II) films.
The electrochemical properties of protoporphyrin IX zinc(II) (ZnPP) films on indium-tin oxide (ITO) substrate have been studied for three types of films with different arrangements, which were an adsorbed film of ZnPP and LB films of ZnPP and its hybrid with hexadecyltrimethylammonium bromide. Cyclic voltammetry (CV) measurement showed that, as the adsorbed amount of ZnPP increases, an irreversible oxidation peak of ZnPP film is intensified. This reveals that electrochemical properties depend on the adsorbed amount rather than the orientation of porphyrin molecules. It was also supported from CV measurement and ultraviolet-visible absorption spectroscopy that porphyrins adsorbed on ITO substrate were desorbed after the single scan of potential. Additionally, photoresponse of these ZnPP films was investigated by photocurrent measurement. The photocurrent generation is due to carboxylic acid moieties but not ZnPP macrocycles.


Iridium-Based Multimetallic Porous Hollow Nanocrystals for Efficient Overall-Water-Splitting Catalysis.
The development of active and durable bifunctional electrocatalysts for overall water splitting is mandatory for renewable energy conversion. This study reports a general method for controllable synthesis of a class of IrM (M = Co, Ni, CoNi) multimetallic porous hollow nanocrystals (PHNCs), through etching Ir-based, multimetallic, solid nanocrystals using Fe3+ ions, as catalysts for boosting overall water splitting. The Ir-based multimetallic PHNCs show transition-metal-dependent bifunctional electrocatalytic activities for both the hydrogen evolution reaction (HER) and oxygen evolution reaction (OER) in acidic electrolyte, with IrCo and IrCoNi PHNCs being the best for HER and OER, respectively. First-principles calculations reveal a ligand effect, induced by alloying Ir with 3d transition metals, can weaken the adsorption energy of oxygen intermediates, which is the key to realizing much-enhanced OER activity. The IrCoNi PHNCs are highly efficient in overall-water-splitting catalysis by showing a low cell voltage of only 1.56 V at a current density of 2 mA cm-2 , and only 8 mV of polarization-curve shift after a 1000-cycle durability test in 0.5 m H2 SO4 solution. This work highlights a potentially powerful strategy toward the general synthesis of novel, multimetallic, PHNCs as highly active and durable bifunctional electrocatalysts for high-performance electrochemical overall-water-splitting devices.


Scalability and cost-effectiveness analysis of whole genome-wide association studies on Google Cloud Platform and Amazon Web Services.
Advancements in human genomics have generated a surge of available data, fueling the growth and accessibility of databases for more comprehensive, in-depth genetic studies. We provide a straightforward and innovative methodology to optimize cloud configuration in order to conduct genome-wide association studies. We utilized Spark clusters on both Google Cloud Platform and Amazon Web Services, as well as Hail (http://doi.org/10.5281/zenodo.2646680) for analysis and exploration of genomic variants dataset. Comparative evaluation of numerous cloud-based cluster configurations demonstrate a successful and unprecedented compromise between speed and cost for performing genome-wide association studies on 4 distinct whole-genome sequencing datasets. Results are consistent across the 2 cloud providers and could be highly useful for accelerating research in genetics. We present a timely piece for one of the most frequently asked questions when moving to the cloud: what is the trade-off between speed and cost?


Sequence of centromere separation: occurrence, possible significance, and control.
This review describes the existence of a phenomenon, sequential separation of centromeres, in mitotic cells of various species including both animals and plants. Critical observations at metaanaphase show that the centromeres of chromosomes in a given genome do not separate into two sister units randomly, but that there is a genetically controlled, nonrandom, species-specific sequence which is independent of the length of the chromosome or the position of the centromere. A stricter control appears to exist for late-separating than for early-separating chromosomes. At early stages of metaanaphase several chromosomes initiate onset of separation simultaneously or in rapid succession, but late-separating chromosomes are better defined in their sequential position. The effect of Colcemid on the sequence of separation is minimal. It is proposed that aneuploidy in humans and other organisms may result from out-of-phase separation of a given chromosome. With the exception of chromosome No. 16, it appears that very early- or very late-separating centromeres are involved in human trisomies more often than those in between. Perhaps one function of centromeric heterochromatin is the control of centromere separation. The amount of such chromatin shows a positive correlation with the timing of separation of the centromeres. Superimposed upon this quantitative influence is the qualitative aspect, as discussed for various genomes. This suggestion explains a lack of extremely large quantities of heterochromatin near the centromere. Its existence in the form of homogeneously staining regions distal to the centromere, as in some cancer cells or in sex chromosomes, seemingly has no influence on the separation of centromeres. A brief discussion of centromere separation errors in human disease is provided, and suggestions for further studies are made.


Capillaroscopic pattern in systemic lupus erythematosus and undifferentiated connective tissue disease: what we still have to learn?
In rheumatology, specific is the capillaroscopic pattern in systemic sclerosis (SSc), the so-called "scleroderma type". Capillaroscopic pattern in systemic lupus erythematosus (SLE) is less specific and includes a wide range of microvascular changes-"SLE-type" capillaroscopic pattern, non-specific findings and in a small percentage "scleroderma-like" pattern. The latter finding is currently associated with a potential subclinical overlap with SSc. Various microvascular changes have been observed in a different proportion of patients with undifferentiated connective tissue disease (UCTD). The aim of the study was to evaluate the capillaroscopic changes in SLE and UCTD. Patients from the following groups were included in the study: 30 female patients with SLE (mean age, 49 ± 15.4 years), 31 patients with UCTD (mean age, 50 ± 17 years; 30 females and 1 male); 34 age- and sex-matched healthy volunteers were examined as a control group. Nailfold capillaroscopy was performed using videocapillaroscope Videocap 3.0 (DS Medica). Capillaroscopic findings were compared with clinical and laboratory data of the patients. At capillaroscopic examination, the most frequent capillaroscopic changes in SLE patients were the presence of elongated capillaries in 43 % (13/30), an increased tortuosity in 70 % (21/30) and a prominent subpapillary plexus in 60 % (18/30) of the cases. In 80 % (24/30) of the patients, dilated capillaries were found; in 6.6 % (2/30), giant capillary loops; and in 16.6 % (5/30), haemorrhages. In 50 % of the patients, an "SLE-type" capillaroscopic pattern was found. In 30 % (9/30) of the cases the capillaroscopic examination revealed "non-specific changes", in 6.6 % (2/30) of the patients it was found a normal capillaroscopic pattern and in 13.3 % (4/30) a "scleroderma-like" pattern. Positive tests for ANA were detected in 73.3 % (11/15) of the patients with "SLE-type" capillaroscopic pattern. In all the patients with "scleroderma-like" capillaroscopic finding, positive autoantibodies with a high titre were found, without signs for overlap with other connective tissue disease (CTD). In two out of four patients with such capillaroscopic findings, a vasculitis of peripheral vessels was evident and in the other two secondary RP and high immunologic activity. A "scleroderma-like" pattern was found in 38 % (12/31) of the patients with UCTD. In 51 % (16/31) of the patients from this group, "non-specific" capillaroscopic findings were observed. For the evaluation of the predictive value of capillaroscopic pattern for the development of a distinct rheumatic disorder in patients with UCTD, a longer period of follow-up is necessary. In SLE patients, it has been found that capillaroscopic examination reveals microvascular changes also in the absence of RP. Here, the results from the study illustrate the correlation between capillaroscopic changes and immunological profile. "Scleroderma-like" capillaroscopic pattern may be observed in the context of active vasculitis of peripheral vessels as well as in patients with secondary RP and high immunologic activity. It does not have an obligatory association with an overlap syndrome with other CTD. Capillaroscopic findings in UCTD are heterogeneous. The potential of capillaroscopic examination in UCTD for evaluating the prognosis of the disease needs to be revealed through long-term follow-up.


A marker of animal-vegetal polarity in the egg of the sea urchin Paracentrotus lividus. The pigment band.
We have examined the subequatorial accumulation of pigment granules (the so-called 'pigment band') in the egg of the sea urchin Paracentrotus lividus, which constitutes an unambiguous marker of animal-vegetal polarity. Most of the reddish pigment granules are situated at the periphery of the egg. They exhibit occasional saltatory movements and can aggregate into large patches. Pigment granules are retained as a band in the isolated cortex when the egg surface complex is isolated by shearing eggs attached to polylysine-coated surfaces with calcium-free isotonic solutions. Pigment granules remain as the main vesicular component of fertilized egg cortices or of unfertilized egg cortices perfused with calcium to provoke cortical granule exocytosis. They may be anchored to the isolated cortex through associations with the plasma membrane and with an extensive subsurface network of rough endoplasmic reticulum (rough ER). Pigment granules contain antimonate-precipitable calcium and, in this respect and many others, resemble acidic vesicles recently identified in the cortex of unpigmented sea urchin eggs. We discuss the similarities observed between granules and acidic vesicles in various urchin egg species and their possible functions.


Variability of treatment selection among surgeons for patients with cT1 urothelial carcinoma.
To report the heterogeneity in the treatment of patients with cT1 urothelial carcinoma by different surgeons, and to report outcomes in patients with and without bacillus Calmette-Guérin (BCG) treatment. We retrospectively reviewed 396 patients who had undergone a re-staging transurethral resection (TUR) for cT1 bladder cancer. We assessed both differences in the treatment by surgeon, and the association of early treatment with BCG with recurrence, progression and bladder cancer-specific death. Muscle was captured in the re-staging TUR specimen in a median of 76% of patients (range 50-94 when stratified by surgeon). On multivariable analysis there was significant heterogeneity among surgeons in the use of early cystectomy (P < 0.001), deferred cystectomy (P < 0.001), and BCG (P= 0.014). However, there was no significant heterogeneity between surgeons in clinical outcome for recurrence (P= 0.9) and overall survival (P= 0.3). Among 288 patients placed on surveillance, the 5-year probability (95% confidence interval) of freedom from recurrence was 45 (36-54)% for those receiving and 54 (44-62)% for those not receiving early BCG. On multivariable analysis, early BCG was not significantly associated with recurrence (P= 0.14). The cumulative incidence of progression was ≈10% for both groups, and the cumulative incidence of bladder cancer-specific death was ≈7% for both groups. The cumulative incidence of deferred cystectomy before progression was 14% for those receiving and 15% for those not receiving early BCG. There is a significant variability among surgeons in the management of patients with T1 disease. The similar outcome for the BCG-treated and -untreated patients in our study is most likely confounded by patient selection.


Correlations between photosynthetic heat tolerance and leaf anatomy and climatic niche in Asian mangrove trees.
Photosynthetic heat tolerance (PHT ) is a key predictor of plant response to climate change. Mangroves are an ecologically and economically important coastal plant community comprised of trees growing at their physiological limits. Mangroves are currently impacted by global warming, yet the PHT of mangrove trees is poorly understood. In this study, we provide the first assessment of PHT in 13 Asian mangrove species, based on the critical temperature that causes the initial damage (TCrit ) and the temperature that causes 50% damage (T50 ) to photosystem II. We tested the hypotheses that the PHT in mangroves is: (i) correlated with climatic niche and leaf traits, and (ii) higher than in plants from other tropical ecosystems. Our results demonstrated correlations between PHT and multiple key climate variables, the palisade to spongy mesophyll ratio and the leaf area. The two most heat-sensitive species were Kandelia obovata and Avicennia marina. Our study also revealed that mangrove trees show high heat tolerance compared to plants from other tropical ecosystems. The high PHT of mangroves thus demonstrated a conservative evolutionary strategy in heat tolerance, and highlights the need for integrative and comparative studies on thermoregulatory traits and climatic niche in order to understand the physiological response of mangrove trees to climate change-driven heatwaves and rising global temperatures.


Early Locomotor Training in Tetraplegic Post-Surgical Dogs with Cervical Intervertebral Disc Disease.
Locomotor training (LT) is task-specific repetitive training, with sensorimotor stimulation and intensive exercises that promote neuromuscular reorganization. This study aimed to observe if LT could be initiated safely in the first 3−15 days after surgery in tetraplegic C1−C5 IVDD—Hansen type I dogs. This prospective blinded clinical study was conducted at two rehabilitation centers in Portugal, with 114 grade 1 (MFS/OFS) dogs, divided by the presence of spinal hyperesthesia into the SHG (spinal hyperesthesia group) (n = 74) and the NSHG (non-spinal hyperesthesia group) (n = 40), evaluated in each time point for two weeks according to a neurorehabilitation checklist by three observers for inter-agreement relation. LT was safely applied with 62.3% of the OFS ≥ 11 within 15 days and of these, 32.4% achieved a OFS ≥ 13. There were no new cases of hyperesthesia in the NSHG and from the SHG all recovered. Comparing groups, a significant difference was observed in their ability to achieve ambulatory status (p < 0.001), between the presence of hyperesthesia and days until ambulation (p < 0.006) and in each time point (p < 0.001; R2 = 0.809). Early LT may be a safe treatment to be applied in the first 3 days on these dogs and spinal hyperesthesia should be important to the rehabilitation team. This study should be continued.


In Vitro and In Silico Analyses of New Cinnamid and Rosmarinic Acid-Derived Compounds Biosynthesized in Escherichia coli as Leishmania amazonensis Arginase Inhibitors.
Arginase is a metalloenzyme that plays a central role in Leishmania infections. Previously, rosmarinic and caffeic acids were described as antileishmanial agents and as Leishmania amazonensis arginase inhibitors. Here, we describe the inhibition of arginase in L. amazonensis by rosmarinic acid analogs (1-7) and new caffeic acid-derived amides (8-10). Caffeic acid esters and amides were produced by means of an engineered synthesis in E. coli and tested against L. amazonensis arginase. New amides (8-10) were biosynthesized in E. coli cultured with 2 mM of different combinations of feeding substrates. The most potent arginase inhibitors showed Ki(s) ranging from 2 to 5.7 μM. Compounds 2-4 and 7 inhibited L. amazonensis arginase (L-ARG) through a noncompetitive mechanism whilst compound 9 showed a competitive inhibition. By applying an in silico protocol, we determined the binding mode of compound 9. The competitive inhibitor of L-ARG targeted the key residues within the binding site of the enzyme, establishing a metal coordination bond with the metal ions and a series of hydrophobic and polar contacts supporting its micromolar inhibition of L-ARG. These results highlight that dihydroxycinnamic-derived compounds can be used as the basis for developing new drugs using a powerful tool based on the biosynthesis of arginase inhibitors.


The effects of coenzyme Q10 on seizures in mice: the involvement of nitric oxide.
Coenzyme Q10 is a potent antioxidant in both mitochondria and lipid membranes. It has also been recognized to have an effect on gene expression. This study was designed to investigate whether acute or subchronic treatment with coenzyme Q10 altered the seizures induced by pentylenetetrazole or electroshock in mice. We also evaluated the involvement of nitric oxide in the effects of coenzyme Q10 in pentylenetetrazole-induced seizure models. Acute oral treatment with different doses of coenzyme Q10 did not affect the seizure in intraperitoneal pentylenetetrazole, intravenous pentylenetetrazole, and electroshock models in mice. Subchronic oral administration of coenzyme Q10 (100 mg/kg or more) increased time latencies to the onset of myoclonic jerks and clonic seizures induced by intraperitoneal pentylenetetrazole and at the doses of 25 mg/kg or more increased the seizure threshold induced by intravenous infusion of pentylenetetrazole. Subchronic doses of coenzyme Q10 (50 mg/kg or more) also decreased the incidence of tonic seizures in the electroshock-induced seizure model. Moreover, acute treatment with the precursor of nitric oxide synthesis, L-arginine (60 mg/kg), led to a significant potentiation of the antiseizure effects of subchronic administration of coenzyme Q10 (400 mg/kg in intraperitoneal and 6.25 mg/kg in intravenous pentylenetetrazole tests). Acute treatment with l-NAME (5 mg/kg), a nonspecific nitric oxide synthase inhibitor, significantly attenuated the antiseizure effects of subchronic doses of coenzyme Q10 in both seizure models induced by pentylenetetrazole. On the other hand, acute administration of aminoguanidine (100 mg/kg), a specific inducible nitric oxide synthase inhibitor, did not affect the seizures in mice treated with subchronic doses of coenzyme Q10 in both intraperitoneal and intravenous pentylenetetrazole tests. In conclusion, only subchronic and not acute administration of coenzyme Q10 attenuated seizures induced by pentylenetetrazole or electroshock. We also demonstrated, for the first time, the interaction between nitric oxide and coenzyme Q10 in antiseizure activity probably through the induction of constitutive nitric oxide synthase.


Fermentation of alginate and its derivatives by different enterotypes of human gut microbiota: Towards personalized nutrition using enterotype-specific dietary fibers.
Alginate and its derivatives are widely used as food additives and dietary fibers. Previous studies indicated that alginate, polyguluronate (PG) and polymannuronate acid (PM) could be fermented by human gut microbiota. However, how different compositions of the microbiota may affect the fermentation outcomes of these polysaccharides remains unknown. Here we show that Bacteroides-dominated microbiota (Bacteroides enterotype) is more proficient at degrading and utilizing PG and PM as compared to Prevotella-dominated (Prevotella enterotype) and Escherichia-dominated microbiota (Escherichia enterotype). Enterotype dictates the fermentation outcomes of the three fibers and the amount of short-chain fatty acids (SCFAs) that are produced. Fermentation of alginate and PM by Bacteroides-dominated microbiota produced the highest amount of total SCFAs and butyrate. Our study demonstrates an enterotype-specific effect of microbiota on the fermentation of alginate and its derivatives and highlights that personalized nutrition using dietary fibers should be tailored according to individual's composition of the gut microbiome.


Tissue imprints in surgical pathology: a rapid intraoperative diagnostic aid.
We studied the intraoperative diagnostic value of imprint cytology in 230 samples obtained from surgical specimens submitted for frozen section diagnosis. A rapid hematoxylin-eosin stain was used. Intraoperative imprint cytology achieved an accuracy rate of 94.3%; for benign lesions the accuracy was 97.5%, and for malignant lesions it was 91%. Overall, the false-negative and suspicious-for-malignancy rates were 1.3% and 4.3%, respectively. No false-positive results were found. The diagnostic yield when intraoperative imprint cytology and frozen section were used together was 99%. It is apparent that imprint cytology is a quick and simple method with wide applicability in the histopathologic diagnosis of lesions from all organs. The value of the method is enhanced when it is used with frozen section diagnosis.


Physiology in perspective: The Wisdom of the Body. Neural control of the kidney.
Cannon equated the fluid matrix of the body with Bernard's concept of the internal environment and emphasized the importance of "the safe-guarding of an effective fluid matrix." He further emphasized the important role of the autonomic nervous system in the establishment and maintenance of homeostasis in the internal environment. This year's Cannon Lecture discusses the important role of the renal sympathetic nerves to regulate various aspects of overall renal function and to serve as one of the major "self-regulatory agencies which operate to preserve the constancy of the fluid matrix."


Modular enzyme assembly for enhanced cascade biocatalysis and metabolic flux.
Enzymatic reactions in living cells are highly dynamic but simultaneously tightly regulated. Enzyme engineers seek to construct multienzyme complexes to prevent intermediate diffusion, to improve product yield, and to control the flux of metabolites. Here we choose a pair of short peptide tags (RIAD and RIDD) to create scaffold-free enzyme assemblies to achieve these goals. In vitro, assembling enzymes in the menaquinone biosynthetic pathway through RIAD-RIDD interaction yields protein nanoparticles with varying stoichiometries, sizes, geometries, and catalytic efficiency. In Escherichia coli, assembling the last enzyme of the upstream mevalonate pathway with the first enzyme of the downstream carotenoid pathway leads to the formation of a pathway node, which increases carotenoid production by 5.7 folds. The same strategy results in a 58% increase in lycopene production in engineered Saccharomyces cerevisiae. This work presents a simple strategy to impose metabolic control in biosynthetic microbe factories.


EEG mismatch responses in a multimodal roving stimulus paradigm provide evidence for probabilistic inference across audition, somatosensation, and vision.
The human brain is constantly subjected to a multimodal stream of probabilistic sensory inputs. Electroencephalography (EEG) signatures, such as the mismatch negativity (MMN) and the P3, can give valuable insight into neuronal probabilistic inference. Although reported for different modalities, mismatch responses have largely been studied in isolation, with a strong focus on the auditory MMN. To investigate the extent to which early and late mismatch responses across modalities represent comparable signatures of uni- and cross-modal probabilistic inference in the hierarchically structured cortex, we recorded EEG from 32 participants undergoing a novel tri-modal roving stimulus paradigm. The employed sequences consisted of high and low intensity stimuli in the auditory, somatosensory and visual modalities and were governed by unimodal transition probabilities and cross-modal conditional dependencies. We found modality specific signatures of MMN (~100-200 ms) in all three modalities, which were source localized to the respective sensory cortices and shared right lateralized prefrontal sources. Additionally, we identified a cross-modal signature of mismatch processing in the P3a time range (~300-350 ms), for which a common network with frontal dominance was found. Across modalities, the mismatch responses showed highly comparable parametric effects of stimulus train length, which were driven by standard and deviant response modulations in opposite directions. Strikingly, P3a responses across modalities were increased for mispredicted stimuli with low cross-modal conditional probability, suggesting sensitivity to multimodal (global) predictive sequence properties. Finally, model comparisons indicated that the observed single trial dynamics were best captured by Bayesian learning models tracking unimodal stimulus transitions as well as cross-modal conditional dependencies.


Exocrine Pancreatic Insufficiency Dosing Guidelines for Pancreatic Enzyme Replacement Therapy Vary Widely Across Disease Types.
Pancreatic enzyme replacement therapy (PERT) is the standard treatment for exocrine pancreatic insufficiency (EPI). However, many individuals are inadequately treated, with gaps in clinical dosing, guidelines, and tools to aid individual titration. A systematic review identified research and guidelines on PERT dosing recommendations across conditions, systematically reviewing and synthesizing total PERT intake, meal/snack guidelines, and changes over time to provide an up-to-date look at the most common doses used in studies and guidelines. This review of 257 articles found wide variability in PERT dosing guidelines within and across conditions. Many patients with EPI are underdosed, with guidelines differing globally and by disease type, and clinician prescribing may also play a role. The most common dosing guidelines focus on starting doses at 40,000-50,000 units of lipase/meal with increases of up to two to three times this amount before pursuing additive therapies. Guidelines and studies typically focus only on fat digestion, and comparison by total daily dose shows underdosing is common. Most PERT studies are on safety and efficacy rather than optimal titration. The current guidelines for PERT in EPI demonstrate substantial variability in dosing recommendations, both within and across disease types. This variation highlights the need for further research to optimize PERT dosing and improve patient outcomes. Healthcare providers should consider individualizing PERT dosing based on nutritional status and response to therapy, ensuring regular follow-up with patients for dose titrations with consideration that most guidelines are framed as initial doses rather than upper limits.


Commercialization of User Data by Developers of Medicines-Related Apps: a Content Analysis.
Developers of medicines-related apps collect a variety of technical, health-related, and identifying user information to improve and tailor services. User data may also be used for promotional purposes. Apps, for example, may be used to skirt regulation of direct-to-consumer advertising of medicines. Researchers have documented routine and extensive sharing of user data with third parties for commercial purposes, but little is known about the ways that app developers or "first" parties employ user data. We aimed to investigate the nature of user data collection and commercialization by developers of medicines-related apps. We conducted a content analysis of apps' store descriptions, linked websites, policies, and sponsorship prospectuses for prominent medicines-related apps found in the USA, Canada, Australia, and UK Google Play stores in late 2017. Apps were included if they pertained to the prescribing, administration, or use of medicines, and were interactive. Two independent coders extracted data from documents using a structured, open-ended instrument. We performed open, inductive coding to identify the range of promotional strategies involving user data for commercial purposes and wrote descriptive memos to refine and detail these codes. Ten of 24 apps primarily provided medication adherence services; 14 primarily provided medicines information. The majority (71%, 17/24) outlined at least one promotional strategy involving users' data for commercial purposes which included personalized marketing of the developer's related products and services, highly tailored advertising, third-party sponsorship of targeted content or messaging, and sale of aggregated customer insights to stakeholders. App developers may employ users' data in a feedback loop to deliver highly targeted promotional messages from developers, and commercial sponsors, including the pharmaceutical industry. These practices call into question developers' claims about the trustworthiness and independence of purportedly evidenced-based medicines information and may create a risk for mis- or overtreatment.


An open volume, high isolation, radio frequency surface coil system for pulsed magnetic resonance.
We present an open volume, high isolation, RF system suitable for pulsed NMR and EPR spectrometers with reduced dead time. It comprises a set of three RF surface coils disposed with mutually parallel RF fields and a double-channel receiver (RX). Theoretical and experimental results obtained with a prototype operating at about 100 MHz are reported. Each surface RF coil (diameter 5.5 cm) was tuned to f0=100.00+/-0.01 MHz when isolated. Because of the mutual coupling and the geometry of the RF coils, only two resonances at f1=97.94 MHz and f2=101.85 MHz were observed. We show they are associated with two different RF field spatial distributions. In continuous mode (CW) operation the isolation between the TX coil and one of the RX coils (single-channel) was about -10 dB. By setting the double-channel RF assembly in subtraction mode the isolation values at f1 or f2 could be optimised to about -75 dB. Following a TX RF pulse (5 micros duration) an exponential decay with time constant of about 600 ns was observed. The isolation with single-channel RX coil was about -11 dB and it increased to about -47 dB with the double-channel RX in subtraction mode. Similar results were obtained with the RF pulse frequency selected to f2 and also with shorter (500 ns) RF pulses. The above geometrical parameters and operating frequency of the RF assembly were selected as a model for potential applications in solid state NMR and in free radical EPR spectroscopy and imaging.


Proximate and ultimate mechanisms of human father-child rough-and-tumble play.
The aim of this contribution is to attempt to understand the adaptive functions of father-child rough-and-tumble play (RTP) in humans. We first present a synthesis of the known proximate and ultimate mechanisms of peer-peer RTP in mammals and compare human parent-child RTP with peer-peer RTP. Next, we examine the possible biological adaptive functions of father-child RTP in humans, by comparing paternal behavior in humans versus biparental animal species, in light of the activation relationship theory and the neurobiological basis of fathering. Analysis of analogies reveals that the endocrine profile of fathers is highly variable across species, compared to that of mothers. This can be interpreted as fathers' evolutionary adjustment to specific environmental conditions affecting the care of the young. Given the high unpredictability and risk-taking features of RTP, we conclude that human adult-child RTP appears to have a biological adaptive function, one of 'opening to the world'.


Increased platelet and red cell counts, blood viscosity, and plasma cholesterol levels during heat stress, and mortality from coronary and cerebral thrombosis.
Recorded deaths from coronary and cerebral thrombosis rise markedly in heat waves. In a British heat wave with little or no distortion due to air-conditioning, outside temperatures of 34.6 degrees C (maximum) and 20.8 degrees C (minimum) were followed by peak mortalities from coronary and cerebral thrombosis one to two days later. Experimental exposure of volunteers to moving air at 41 degrees C for six hours caused core temperature to rise 0.84 degree C, weight to fall 1.83 kg with sweating despite access to water, heart rate to increase 32 beats per minute, and arterial pressure to fall, particularly on standing. The red blood cell count increased 9 percent, and blood viscosity increased 24 percent, mostly after the first hour. The platelet count rose 18 percent, and the platelet volume fell, mostly in the first hour. The plasma cholesterol level increased 14 percent without a change in distribution among lipoprotein fractions. The changes seem able to explain the increased mortality from arterial thrombosis in hot weather.


Biomimetic Nanoerythrosome-Coated Aptamer-DNA Tetrahedron/Maytansine Conjugates: pH-Responsive and Targeted Cytotoxicity for HER2-Positive Breast Cancer.
DNA materials have emerged as potential nanocarriers for targeted cancer therapy to precisely deliver cargos with specific purposes. The short half-life and low bioavailability of DNA materials due to their interception by the reticuloendothelial system and blood clearance further limit their clinical translation. This study employs an HER2-targeted DNA-aptamer-modified DNA tetrahedron (HApt-tFNA) as a drug delivery system, and combines maytansine (DM1) to develop the HApt-DNA tetrahedron/DM1 conjugate (HApt-tFNA@DM1, HTD, HApDC) for targeted therapy of HER2-positive cancer. To optimize the pharmacokinetics and tumor-aggregation of HTD, a biomimetic camouflage is applied to embed HTD. The biomimetic camouflage is constructed by merging the erythrocyte membrane with pH-responsive functionalized synthetic liposomes, thus with excellent performance of drug delivery and tumor-stimulated drug release. The hybrid erythrosome-based nanoparticles show better inhibition of HER2-positive cancer than other drug formulations and exhibit superior biosafety. With the strengths of precise delivery, increased drug loading, sensitive tumor probing, and prolonged circulation time, the HApDC represents a promising nanomedicine to treat HER2-positive tumors. Notably, this study developsa dual-targeting nanoparticle by combining pH-sensitive camouflage and HApDC, initiating an important step toward the development and application of DNA-based medicine and biomimetic cell membrane materials in cancer treatment and other potential biological applications.


Broccoli (Brassica oleracea L. var. italica) cultivars, Palam Samridhi and Palam Vichitra affect the growth of Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae).
Effect of the ethyl acetate seed extracts of two cultivars of broccoli, Brassica oleracea Italica, Palam Samridhi (PS) and Palam Vichitra (PV) on growth, development and nutritional physiology of an economically important insect pest, Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae) was evaluated by conducting bioassays and nutritional assays. The insect larvae were fed on diets amended with the seed extracts of two cultivars at different concentrations viz. 5, 25, 125, 625 and 3125 ppm and taking water as control. The response of the insect varied with plant varieties. The extracts disrupted the developmental cycle of the pest. Larval mortality and total adult emergence were negatively affected. Larval period and total development period were also negatively influenced. Nutritional indices of S. litura also showed considerable decrease in the RGR, RCR, ECI and ECD as compared to control. The AD values were also enhanced with both the cultivars. The findings of the study revealed a considerable anti-insect potential of the two extracts and needs to be further explored for identification and isolation of bioactive constituents from broccoli for efficient management of the pest population.


Engineered macromolecular Toll-like receptor agents and assemblies.
Macromolecular Toll-like receptor (TLR) agents have been utilized as agonists and inhibitors in preclinical and clinical settings. These agents interface with the TLR class of innate immune receptors which recognize macromolecular ligands that are characteristic of pathogenic material. As such, many agents that have been historically investigated are derived from the natural macromolecules which activate or inhibit TLRs. This review covers recent research and clinically available TLR agents that are macromolecular or polymeric. Synthetic materials that have been found to interface with TLRs are also discussed. Assemblies of these materials are investigated in the context of improving stability or efficacy of ligands. Attention is given to strategies which modify or enhance the current agents and to future outlooks on the development of these agents.


Quantification of neocortical slice diffusion characteristics using pharmacokinetic and pharmacodynamic modelling.
Pharmacological brain slice experiments are complicated by the need to ensure adequate drug delivery deep into the healthy layers of the tissue. Because tissue slices have no blood supply, this is achieved solely by passive drug diffusion. The aim of this study was to determine whether pharmacokinetic/pharmacodynamic (PKPD) modeling could be adapted to estimate drug diffusion times in neocortical brain slices. No-magnesium seizure-like event (SLE) activity was generated in 41 slices (400 μ m). Two anesthetic agents, etomidate (24 μ M, n = 14) and thiopental (250 μ M, n = 14), and magnesium ions (n = 13) were delivered to effect reversible reductions in SLE frequency. Concentration-effect hysteresis loops were collapsed using a first order rate constant model and equilibrium half-lives (t1/2Ke0) derived. The t1/2Ke0 values obtained were consistent with expectations. The median (range) t1/2Ke0 of 83.1 (19.4-330.1) min for etomidate is in keeping with its known slow diffusion into brain slice tissue. Values for etomidate and thiopental (111.8 (27.8-198.0) min) were similar, while magnesium had a significantly faster equilibration rate (t1/2Ke0 of 26.1 (8.6-77.0) min) compared to the anesthetics, as expected for a simple ion. In conclusion, PKPD modeling is a simple and practical method that can be applied to brain slice experiments for investigating drug diffusion characteristics.


Safety evaluation of mycotoxin citrinin production from Monascus ruber through whole-genome sequencing and analytical evaluation.
In this study, the whole genome of Monascus ruber KACC 46666 was generated using the PacBio RSII sequencer with high-quality de novo assembly to obtain trustworthy assembly and annotation using genome assemblies with long reads from PacBio single-molecule real-time sequencing. The whole genome of M. ruber has a total length of 25.9 Mb, divided in 13 contigs with 9639 genes. The functions of genes involved in secondary metabolite production were further analyzed. Gene clusters involved in the production of Monascus pigment, monacolin K, and mycotoxin citrinin were identified. Notably, most of the citrinin gene cluster was lost, as confirmed via high-performance liquid chromatography analysis. This genome-level safety evaluation of industrially important Monascus strains will provide valuable information for genome-based microbial engineering of natural food colorants and production of commercially important secondary metabolites such as monacolin K.


Crystal structure of T4 endonuclease VII resolving a Holliday junction.
Holliday proposed a four-way DNA junction as an intermediate in homologous recombination, and such Holliday junctions have since been identified as a central component in DNA recombination and repair. Phage T4 endonuclease VII (endo VII) was the first enzyme shown to resolve Holliday junctions into duplex DNAs by introducing symmetrical nicks in equivalent strands. Several Holliday junction resolvases have since been characterized, but an atomic structure of a resolvase complex with a Holliday junction remained elusive. Here we report the crystal structure of an inactive T4 endo VII(N62D) complexed with an immobile four-way junction with alternating arm lengths of 10 and 14 base pairs. The junction is a hybrid of the conventional square-planar and stacked-X conformation. Endo VII protrudes into the junction point from the minor groove side, opening it to a 14 A x 32 A parallelogram. This interaction interrupts the coaxial stacking, yet every base pair surrounding the junction remains intact. Additional interactions involve the positively charged protein and DNA phosphate backbones. Each scissile phosphate that is two base pairs from the crossover interacts with a Mg2+ ion in the active site. The similar overall shape and surface charge potential of the Holliday junction resolvases endo VII, RuvC, Ydc2, Hjc and RecU, despite having different folds, active site composition and DNA sequence preference, suggest a conserved binding mode for Holliday junctions.


Impact of microplastics on the foraging, photosynthesis and digestive systems of submerged carnivorous macrophytes under low and high nutrient concentrations.
The prevalence of microplastics in marine, terrestrial, and freshwater habitats has raised concerns about their availability and risks to organisms. However, the effects of plastic debris on aquatic plants remain largely unknown and have hardly been studied, despite the importance of these plants in freshwater ecosystems. In this study, we exposed the aquatic carnivorous plant Utricularia vulgaris to polystyrene microplastics (10 and 80 mg/L) combined with different nutrient concentrations and then assessed their effects on the growth rate, photosynthesis, oxidative stress, and trap-associated microbial community of U. vulgaris. The impact of microplastic accumulation in the traps (or "bladders") of U. vulgaris was investigated using confocal microscopy. The results showed that the relative growth rate, shoot length, chlorophyll content, Fv/Fm, and ascorbate peroxidase enzyme activity of U. vulgaris decreased in 80 mg/L microplastics, whereas the superoxide dismutase and peroxidase enzyme activities increased significantly. The presence of microplastics led to higher malondialdehyde and hydrogen peroxide contents. However, high nutrient concentrations can compensate for the reduced growth performance of U. vulgaris in microplastic-exposure treatments. The microplastic treatments significantly altered the trap-associated microbial community structure and diversity. The results of this study revealed that beside adsorption, carnivorous plants can accumulate microplastics in their digestive organs.


Neonatal retrievals from a regional centre: Outcomes, missed opportunities and barriers to back transfer.
High-risk neonates are retrieved from regional centres to tertiary neonatal units when the required care of the baby exceeds the clinical capabilities of the birthing facility. However, there is limited research on the outcomes of neonatal retrievals from regional special care centres and the barriers to back transfer of neonates from a tertiary centre are not well established. This study aimed to review the outcome of neonatal retrievals >32 weeks gestation from a regional referral centre. The study also aimed to determine missed opportunities for providing care at the regional centre and evaluate patient back transfer delays. All neonates transferred to a tertiary neonatal intensive care unit in North Queensland over the 5-year period January 2016 to December 2020 from a regional neonatal centre were retrospectively reviewed from the electronic medical records. Fifty neonates transferred to a tertiary neonatal intensive care unit over the study period were identified. Between 2016 and 2020, the number of neonatal retrievals increased (P = 0.021). Out of the 50 neonatal retrievals, 86% were for medical reasons. Overall, eight neonates were identified as missed opportunities whose care could have been maintained at the regional centre with support from the tertiary neonatal intensive care unit. In total, 16 neonates were affected by a delay in back transfer. This study shows a significant increase in retrievals to tertiary neonatal intensive care unit over the study period. Increasing bed capacity, utilising telehealth and recruiting regional special care nursery staff could improve outcomes and reduce strain on tertiary neonatal resources.


Haemopoietic growth factor production by normal and aplastic anaemia stroma in long-term bone marrow culture.
Defective marrow stroma, or microenvironment, have been proposed as one of several mechanisms to account for bone marrow failure in aplastic anaemia (AA). This could involve defects in positive- or negative-acting haemopoietic regulator expression by AA stroma, or alteration of normal stroma-stem cell interactions. We have used a sensitive bioassay to investigate production of granulocyte-colony stimulating factor (G-CSF), granulocyte-macrophage-colony stimulating factor (GM-CSF), interleukin (IL)-3, IL-6 and stem cell growth factor (SCF), by normal and AA stroma in long-term bone marrow culture (LTBMC). LTBMC were grown to confluence, irradiated and harvested to yield a single cell suspension. These cells were cocultured with normal target bone marrow mononuclear cells (BMMC), or CD34+ cells, in clonogenic assays, in the absence of exogenous cytokines. Cytokines responsible for the colony-stimulating activity (CSA) and burst-promoting activity (BPA) produced by stromal cells were identified by neutralizing antibodies to specific cytokines. All normal stroma populations produced G-CSF and GM-CSF, 93% produced IL-3, 80% produced IL-6, and 70% produced SCF. Similarly, all AA stroma produced G-CSF and GM-CSF, and 71% produced SCF. In contrast, only 71% of AA stroma produced IL-3 and 36% produced IL-6. Target cell stimulation was not dependent on direct stroma-target cell contact, suggesting production of soluble cytokines. However, although both IL-6 and G-CSF were detected in LTBMC supernatants by enzyme-linked immunoassay (ELISA), IL-3 and GM-CSF were undetectable, perhaps indicating low-level local production of these factors.


Functional analysis of the conserved hydrophobic gate region of the magnesium transporter CorA.
The Leu294 residue in the cytoplasmic neck of Thermotoga maritima CorA is considered to be the main gate for Mg2+ transport. We created three site-directed mutants at this position: in the Leu294Asp and Leu294Gly mutants we observed a defect in closing of the pore, while in the Leu294Arg mutant not only gating, but also the regulation of Mg2+ uptake was affected. Our results confirmed the importance of the Leu294 for gating of Mg2+ transport and in addition revealed the influence of the charge and structural features of the amino acid residues on the gating mechanism.


On the inhibition of 5-lipoxygenase product formation by tryptanthrin: mechanistic studies and efficacy in vivo.
Leukotrienes (LTs) are pro-inflammatory mediators produced by 5-lipoxygenase (5-LO). Currently available 5-LO inhibitors either lack efficacy or are toxic and novel approaches are required to establish a successful anti-LT therapy. Here we provide a detailed evaluation of the effectiveness of the plant-derived alkaloid tryptanthrin as an inhibitor of LT biosynthesis. We analysed LT formation and performed mechanistic studies in human neutrophils stimulated with pathophysiologically relevant stimuli (LPS and formyl peptide), as well as in cell-free assays (neutrophil homogenates or recombinant human 5-LO) and in human whole blood. The in vivo effectiveness of tryptanthrin was evaluated in the rat model of carrageenan-induced pleurisy. Tryptanthrin potently reduced LT-formation in human neutrophils (IC(50) = 0.6µM). However, tryptanthrin is not a redox-active compound and did not directly interfere with 5-LO activity in cell-free assays. Similarly, tryptanthrin did not inhibit the release of arachidonic acid, the activation of MAPKs, or the increase in [Ca(2+) ](i) , but it modified the subcellular localization of 5-LO. Moreover, tryptanthrin potently suppressed LT formation in human whole blood (IC(50) = 10µM) and reduced LTB(4) levels in the rat pleurisy model after a single oral dose of 10mg·kg(-1) . Our data reveal that tryptanthrin is a potent natural inhibitor of cellular LT biosynthesis with proven efficacy in whole blood and is effective in vivo after oral administration. Its unique pharmacological profile supports further analysis to exploit its pharmacological potential.


[Standard wrist arthroscopy: technique and documentation].
Minimally invasive approach to the wrist in order to diagnose and treat different wrist pathologies. Diagnosis of unclear chronic pain syndromes, cartilage status, intra-articular ligament structures as well as post-traumatic and inflammatory conditions of the wrist. Treatment of ulnar impaction syndrome, dorsal ganglia and also in fracture treatment and various different wrist interventions. Soft tissue infections around the wrist, severe scarring may impede access to the joint. Supine position with the forearm upright and in neutral position, the elbow flexed by 90°, axial traction of 3-4 kg. Superficial stab incision, blunt preparation through joint capsule, insertion of optic through 3-4 portal and probe through 4-5 portal, and radial and ulnar midcarpal portals, respectively. Either sodium chloride, CO2 or air is used as arthroscopy medium. Diagnostic round with standardized examination of all parts of the joint. Standardized written and high quality photo and/or video documentation to facilitate understandability of the findings. Forearm cast for 1 week, limited load for 2 more weeks, then load is increased to normal.


Taxonomy of Euschizomerus Chaudoir 1850 (Coleoptera: Carabidae: Panagaeinae) from India with a new species and new synonym.
Description of new species Euschizomerus devagiriensis sp. nov. from south India, synonymisation of Euschizomerus schuhi Kirschenhofer, 2000 with E. indicus Jedlicka, 1955, redescriptions and new records of the earlier reported Indian species with a key to the species are provided.


Short sleep duration is associated with increased obesity markers in European adolescents: effect of physical activity and dietary habits. The HELENA study.
Adequate sleep is a critical factor for adolescent's health and health-related behaviors. (a) to describe sleep duration in European adolescents from nine countries, (b) to assess the association of short sleep duration with excess adiposity and (c) to elucidate if physical activity (PA), sedentary behaviors and/or inadequate food habits underlie this association. A sample of 3311 adolescents (1748 girls) aged 12.5-17.49 years from 10 European cities in Austria, Belgium, France, Germany, Greece, Hungary, Italy, Spain and Sweden was assessed in the Healthy Lifestyle in Europe by Nutrition in Adolescence Study between 2006 and 2008. We measured anthropometric data, sleep duration, PA (accelerometers and questionnaire), television watching and food habits (Food Frequency Questionnaire). Average duration of daily sleep was 8 h. Shorter sleepers showed higher values of BMI, body fat, waist and hip circumferences and fat mass index (P<0.05), particularly in females. Adolescents who slept <8 h per day were more sedentary, as assessed by accelerometry, and spent more time watching TV (P<0.05). The proportion of adolescents who eat adequate amounts of fruits, vegetables and fish was lower in shorter sleepers than in adolescents who slept ≥8 h per day, and so was the probability of having adequate food habits (P<0.05). Correlation analysis indicated that short sleep is associated with higher obesity parameters. In European adolescents, short sleep duration is associated with higher adiposity markers, particularly in female adolescents. This association seems to be related to both sides of the energy balance equation due to a combination of increased food intake and more sedentary habits.