Identification of a plasmin-interactive site within the A2 domain of the factor VIII heavy chain.
Factor VIII is activated and inactivated by plasmin by limited proteolysis. In our one-stage clotting assay, these plasmin-catalyzed reactions were inhibited by the addition of isolated factor VIII A2 subunits and by Glu-Gly-Arg-active-site modified factor IXa. SDS-PAGE analysis showed that an anti-A2 monoclonal antibody, recognizing the factor IXa-interactive site (residues 484-509), blocked the plasmin-catalyzed cleavage at Arg(336) and Arg(372) but not at Arg(740). Surface plasmon resonance-based assays and ELISA demonstrated that the A2 subunit bound to active-site modified anhydro-plasmin with high affinity (K(d): 21 nM). Both an anti-A2 monoclonal antibody and a peptide comprising of A2 residues 479-504 blocked A2 binding by approximately 80% and approximately 55%, respectively. Mutant A2 molecules where the basic residues in A2 were converted to alanine were evaluated for binding of anhydro-plasmin. Among the tested mutants, the R484A A2 mutant possessed approximately 250-fold lower affinity than the wild-type A2. The affinities of K377A, K466A, and R471A mutants were decreased by 10-20-fold. The inhibitory effect of R484A mutant on plasmin-catalyzed inactivation of factor VIIIa was approximately 20% of that of wild-type A2. In addition, the inactivation rate by plasmin of factor VIIIa reconstituted with R484A mutant was approximately 3-fold lower than that with wild-type A2. These findings demonstrate that Arg(484) plays a key role within the A2 plasmin-binding site, responsible for plasmin-catalyzed factor VIII(a) inactivation.


Dissection of the antibacterial mechanism of zinc oxide nanoparticles with manipulable nanoscale morphologies.
Despite the extensive uses of ZnO nanoparticles as promising antimicrobial agents to tackle the severe microbial infections, the systematic antibacterial studies on ZnO nanoparticles with manipulable nanoscale morphologies at the genetic expression level remain ill-defined. In this study, via a controllable thermal decomposition, ZnO nanoparticles of different morphologies were facilely prepared. Additionally, the surface PEGylation of ZnO was conducted to obtain the nanoparticles of low biotoxicity. While all the prepared ZnO nanoparticles exhibited the significantly chemical activities, the pronounced antibacterial effect of obtained ZnO nanoparticles was also identified, in which the ultra-small ones (~5 nm) showed the best performance. Moreover, the antibacterial activities of ZnO nanoparticles were studied by bacterial nucleic acid leakage, alkaline phosphatase, biofilm and reactive oxygen species (ROS) assays. Furthermore, the transcriptome analysis of ZnO nanoparticles with different morphologies against Escherichia coli (E. coli) revealed the underlying antibacterial mechanism involved the signal transduction, material transport, energy metabolism and other biological processes. Therefore, the cost-effective preparation of ZnO nanoparticles with distinct morphological features provides insights for the development of application specific antibacterial agents.


Neohemocytes.
The neohemocytes described in this report average approximately 0.4 microns in diameter. The microcapsule membrane is composed of biodegradable lipids, including phospholipids, and has a structure similar to liposomes. In a suspension having an apparent hematocrit of 0.5, the neohemocytes occupy 50% of the volume, the neohemocyte membrane accounts for approximately 2%, and the hemoglobin suspension accounts for about 48% of the volume. The encapsulated hemoglobin suspension averages 15.8g%. The P50 averages 26, the Hill Number averages 2.1, and methemoglobin is typically less than 5%. Transfusions in rats of neohemocyte suspensions, where the RBC hematocrit is lowered below 0.03, consistently give a fivefold-or-better increase in survival time relative to transfusions of equal concentrations of unencapsulated hemoglobin. Many of the problems of hemoglobin microencapsulation have been overcome, and the results strongly indicate that neohemocytes may become the functional component of a resuscitative fluid for use in man.


Utility of chromogranin A, pancreatic polypeptide, glucagon and gastrin in the diagnosis and follow-up of pancreatic neuroendocrine tumours in multiple endocrine neoplasia type 1 patients.
Pancreatic neuroendocrine tumours (PNETs) are the major source of disease-specific mortality in multiple endocrine neoplasia type 1 (MEN1) patients. Chromogranin A (CgA), pancreatic polypeptide (PP), glucagon and gastrin have some diagnostic value in sporadic PNETs, but there is very little evidence for their efficacy in diagnosing PNETs in MEN1 patients. We performed a retrospective chart review of the existing MEN1 database in our institution. One hundred and thirteen patients were eligible for diagnostic value analysis of tumour markers. Patients were excluded if measurement of tumour markers was missing, either 3 months prior to PNET diagnosis (PNET patients) or prior to abdominal imaging (non-PNET patients). Clinicopathologic characteristics and of tumour marker measurements were analysed. Of 293 confirmed MEN1 cases, 55 PNETs and 58 non-PNETs met inclusion criteria. The area under the curve (AUC) for CgA, PP, glucagon and gastrin in MEN1 cases was 59·5%, 64·1%, 77·0% and 75·9%, respectively. The AUC for the combination of CgA, PP and gastrin was 59·6%. PP, but not CgA, glucagon or gastrin was significantly associated with both age and PNET functional status (P = 0·0485 and 0·0188, respectively). No markers were significantly associated with sex, PNET size, tumour number, tumour location, American Joint Committee on Cancer (AJCC) stage, presence of lymph node metastasis, lymphovascular invasion or overall survival. CgA values were not significantly lower following PNET resection than pre-operatively (P = 0·554). The value of blood markers for diagnosing PNETs in MEN1 patients is relatively low, even when used in combination.


A new risk predictive scoring system of vasovagal reactions in patients with preoperative autologous blood donation.
Vasovagal response (VVR) is the most common adverse reaction during blood donation and it is the main element for the safety of the patients with preoperative autologous blood donation (PABD). Accurate identification high-risk group is of great significance for PABD. Our study aimed to establish a scoring system based on the nomogram to screen the high-risk population and provide evidence for preventing the occurrence of VVRs. A number of 4829 patients underwent PABD between July 2017 and June 2020 in the first medical center of Chinese PLA Hospital were recruited, 3387 of whom were included in the training group (70 %; 108 VVRs patients vs 3279 Non-VVRs patients), 1442 were included in the validation group (30 %; 46 VVRs patients vs 1396 Non-VVRs patients). The data were analyzed by univariate and multivariate logistic regression. The nomogram of the scoring system was created by using the RMS tool in R software. Seven variables including BMI, hematocrit, pre-phlebotomy heart rate and systolic blood pressure, history of blood donation, age group and primary disease were selected to build the nomogram, which was shown as prediction model. And the score was 0-1 for BMI, 0-2 for hematocrit, systolic blood pressure, heart rate and no blood donation history, 0-10 for age, 0-3 for primary disease. When the total cutoff score was 11, the predictive system for identifying VVRs displayed higher diagnostic accuracy. The area under the curve, specificity, and sensitivity of the training group were 0.942, 82.41 % and 97.17 %, respectively, whereas those of the validation group were 0.836, 78.26 % and 78.15 %, respectively. A risk predictive scoring system was successfully developed to identify high-risk VVRs group form PABD patients that performed well.


The role of endoscopic doppler probe in the management of bleeding peptic ulcers: a systematic review and meta-analysis.
Forrest classification for ulceration has significant intra and inter-observer variability. The endoscopic doppler probe (DOP-US) identifies arterial blood flow at the base to direct therapy. We performed a systematic review and meta-analysis to evaluate the role of the DOP-US in bleeding peptic ulcers. Three independent reviewers performed a comprehensive review of all original articles published from inception to December 2019, evaluating the use of DOP-US in peptic ulcer bleeding. Primary outcomes were the comparison of rebleeding rate, mortality, and surgical intervention in patients with DOP-US signal-guided therapy versus standard visual evaluation guided therapy. Eight studies were included after a thorough search was concluded using the key words. The use of DOP-US probe decreases rebleeding, mortality, and surgical intervention as compared to Forrest Classification. The risk of rebleeding is significantly higher if the signal persists despite endoscopic therapy (48.5% (95% CI 29.5-67.9%)). The first systematic review and meta-analysis showed that the DOP-US is a beneficial tool in the management of bleeding ulcers and adds valuable information to visual evaluation.


Strategies to minimize placebo effects in research investigations.
Placebo-controlled trials are the research standard to evaluate new interventions for which there is no standard of care. While lessening performance and detection bias, such design provides a direct mode of comparison against the probed intervention. Still, using placebo arms may pose new challenges to the design, conduct and analysis of clinical trials. This is particularly relevant in circumstances of non-additivity between the therapeutic and the placebo effects, if the outcome of interest has floor or ceiling effects, or when the predicted effect size of the intervention is large and leads to small sample sizes. There are several possible strategies to mitigate the confounding effects of the placebo, each relevant to specific clinical trial designs. This chapter puts into context the new challenges created by the placebo effect, discusses possible ways around them, and explores the future of the field.


Sex-specific myoelectric manifestations of localized fatigue during a multi-joint repetitive task.
We have previously demonstrated that fatigue at different locations impacts joint angles, angular variability, and coordination variability differently. However, the neuromuscular control aspects underlying these kinematic changes have never been demonstrated. Seventeen young adults (8 males) were recruited. Electromyographic electrodes were placed on: upper trapezius, pectoralis major, anterior and middle deltoid, biceps and triceps brachii, and left and right erector spinae. Subjects performed the repetitive pointing task (RPT) at 1 Hz for 30 s before and after localized fatigue tasks, which consisted of one shoulder, one elbow and one lower back isometric fatiguing protocols until exhaustion in randomized order. Electromyographic amplitude (RMS), variability (SD) and mean power frequency (MnPF) were calculated for each of the pre-fatigue and post-fatigue RPT trials. There were sex × fatigue location interaction effects on upper trapezius RMS (p = 0.038) with males' values increasing the most after shoulder fatigue. Females' triceps brachii RMS was greater compared to males after shoulder, elbow, and trunk fatigue (p = 0.003, p = 0.001 and p = 0.007 respectively). There were sex × fatigue location effects on left erector spinae MnPF (p = 0.011) with males and females' values decreasing the most after trunk fatigue, but more so in males. Results demonstrate that males and females compensate differently during a repetitive pointing task when their elbows, shoulders and trunks are locally fatigued, which could have implications on sex-specific workplace injury risks. See Table 1 for acronyms.


Sustained housing-type social buffering following social housing in male rats.
In social animals, recovery from the adverse effects of distressing stimuli is promoted by subsequent cohousing with a conspecific animal(s). This phenomenon has been termed housing-type social buffering. We previously found that social housing induced housing-type social buffering in fear-conditioned male rats. This buffering took the form of attenuated conditioned hyperthermia in response to an auditory conditioned stimulus (CS). Here, we assessed whether this social buffering is sustained even if the subject is housed alone after a period of social housing. When fear-conditioned subjects were housed alone during a 48-h period between conditioning and re-exposure to the auditory CS, they exhibited conditioned hyperthermia in response to the CS. However, conditioned hyperthermia was not observed when the 12-h period of social housing began 24 and 36h after conditioning during the 48-h period. This was not the case when the 12-h period of social housing began 0 and 12h after the conditioning. These results suggest that housing-type social buffering is sustained for 12h after the 12-h period of social housing. We next considered whether increasing the duration of social housing would extend the period of social buffering. We observed social buffering of conditioned hyperthermia 24 and 48, but not 96h after a 24-h period of social housing. These results suggest that social buffering was extended when the duration of social housing was increased. Taken together, our findings indicate that housing-type social buffering is sustained after a period of social housing.


Depression, anxiety, and stress and socio-demographic correlates among general Indian public during COVID-19.
The severe outbreak of COVID-19 has affected the mental health of Indians. The objective of this article was to find the prevalence rates of depression, anxiety and stress and their socio-demographic correlates among Indian population during the lockdown to contain the spread of COVID-19. A cross-sectional survey was conducted using an electronic questionnaire. A total of 354 participants were recruited through convenience sampling. Depression, anxiety and stress were measured using Depression Anxiety Stress Scale (DASS-21), a 21-item self-reported questionnaire. In total, 25%, 28% and 11.6% of the participants were moderate to extremely severely depressed, anxious and stressed, respectively. Binary logistic regressions indicated employment status (odds ratio (OR) = 1.91; 95% confidence interval (CI): 1.072-3.418) and binge drinking (OR = 2.03; 95% CI: 1.045-3.945) were significantly associated with depressive symptoms; gender (OR = 2.17; 95% CI: 1.317-3.589), employment status (OR = 1.77; 95% CI: 1.002-3.141) and binge drinking (OR = 2.62; 95% CI: 1.361-5.048) were significantly associated with anxiety symptoms; and binge drinking (OR = 3.42; 95% CI: 1.544-7.583) was significantly associated with stress symptoms. Depression, anxiety and stress among Indian population during the lockdown were prevalent. Along with other measures to contain the spread of COVID-19, mental health of citizens needs the urgent attention of the Indian government and mental health experts. Further large-scale studies should be conducted on different professions and communities such as health care professionals and migrant workers and incorporate other mental health indicators.


Anti-inflammatory activity of hydroalcoholic extracts of Lavandula dentata L. and Lavandula stoechas L.
Plants from genus Lavandula have been used as anti-inflammatory drugs in Mediterranean traditional medicine. Nowadays, there is a growing interest for complementary medicine, including herbal remedies, to treat inflammatory bowel disease (IBD). To test the anti-inflammatory properties of Lavandula dentata and Lavandula stoechas extracts in two inflammatory experimental models: TNBS model of rat colitis and the carrageenan-induced paw edema in mice, in order to mimic the intestinal conditions and the extra-intestinal manifestations of human IBD, respectively. The extracts were characterized through the qualitative HPLC analysis. Then, they were assayed in vitro and in vivo. In vitro studies were performed in BMDMs and CMT-93 epithelial cells with different concentrations of the extracts (ranging from 0.1 to 100µg/ml). The extracts were tested in vivo in the TNBS model of rat colitis (10 and 25mg/kg) and in the carrageenan-induced paw edema in mice (10, 25 and 100mg/kg). L. dentata and L. stoechas extracts displayed immunomodulatory properties in vitro down-regulating different mediators of inflammation like cytokines and nitric oxide. They also showed anti-inflammatory effects in the TNBS model of colitis as evidenced by reduced myeloperoxidase activity and increased total glutathione content, indicating a decrease of neutrophil infiltration and an improvement of the oxidative state. Besides, both extracts modulated the expression of pro-inflammatory cytokines and chemokines, and ameliorated the altered epithelial barrier function. They also displayed anti-inflammatory effects in the carrageenan-induced paw edema in mice, since a significant reduction of the paw thickness was observed. This was associated with a down-regulation of the expression of different inducible enzymes like MMP-9, iNOS and COX-2 and pro-inflammatory cytokines, all involved in the maintenance of the inflammatory condition. L. dentata and L. stoechas extracts showed intestinal anti-inflammatory effect, confirming their potential use as herbal remedies in gastrointestinal disorders. In addition, their anti-inflammatory effect was also observed in other locations, thus suggesting a possible use for the treatment of the extra-intestinal symptoms of IBD.


Development and experimental medicine applications of PET in oncology: a historical perspective.
Nearly 90 years of scientific research have led to the use of PET and the ability to forge advances in the field of oncology. In this Historial Review we outline the key developments made with this imaging technique, including the evolution of cyclotrons and scanners, together with the associated advances made in image reconstruction, presentation, analysis of data, and radiochemistry. The applications of PET to experimental medicine are summarised, and we cover how these are related to the use and development of PET, especially in the assessment of tumour biology and pharmacology. The use of PET in clinical oncology and for tissue pharmacokinetic and pharmacodynamic studies as a means of supporting drug development is detailed. The current limitations of the technology are also discussed.


Selective depletion and reconstitution of nucleosome core particles.
A method for the complete and specific removal of histones H2A and H2B from nucleosome core particles is presented. Reconstitution of the separated products of depletion form a particle which has the same structure as native core particles as judged by a number of physical and biochemical criteria. The technique described also minimises the possibility of the formation of reconstituted core particles with different histone stoichiometries. These experiments are important as they demonstrate a procedure which can be extended to prepare core particles with selectively deuterated components while maintaining complete integrity of structure. When prepared, and studied by neutron scattering, selectively-deuterated core particles can give detailed information with respect to the relative positions and structure of the histone fractions within the core particle.


Diastereoselective synthesis of optically active (2 R,5 R)-hexanediol.
Diastereoselective reduction of diketones with Lactobacillus kefir DSM 20587 was examined. The reduction of both oxo-functions proceeded highly diastereoselectively. (2 R,5 R)-Hexanediol 3 was produced starting from (2,5)-hexanedione 1 in quantitative yields with enantiomeric excess >99% and diastereomeric excess >99%. The reaction conditions were optimized: maximum yield of (2 R,5 R)-hexanediol was reached at pH 6, 30 degrees C and with equal amounts of substrate and cosubstrate. The applicability of the system in fed-batch experiments was demonstrated. The feed specific biomass concentration required to reach maximal yield and selectivity in fed-batch mode was determined.


EGFR Mutations in GBM Alter Ligand Binding and Subsequent Dimerization.
Mutations in EGFR in glioblastoma (GBM) affect its ability to discriminate between activating ligands.


Limited Effect of Rebamipide in Addition to Proton Pump Inhibitor (PPI) in the Treatment of Post-Endoscopic Submucosal Dissection Gastric Ulcers: A Randomized Controlled Trial Comparing PPI Plus Rebamipide Combination Therapy with PPI Monotherapy.
The ability of endoscopic submucosal dissection (ESD) to resect large early gastric cancers (EGCs) results in the need to treat large artificial gastric ulcers. This study assessed whether the combination therapy of rebamipide plus a proton pump inhibitor (PPI) offered benefits over PPI monotherapy. In this prospective, randomized, multicenter, open-label, and comparative study, patients who had undergone ESD for EGC or gastric adenoma were randomized into groups receiving either rabeprazole monotherapy (10 mg/day, n=64) or a combination of rabeprazole plus rebamipide (300 mg/day, n=66). The Scar stage (S stage) ratio after treatment was compared, and factors independently associated with ulcer healing were identified by using multivariate analyses. The S stage rates at 4 and 8 weeks were similar in the two groups, even in the subgroups of patients with large amounts of tissue resected and regardless of CYP2C19 genotype. Independent factors for ulcer healing were circumferential location of the tumor and resected tissue size; the type of treatment did not affect ulcer healing. Combination therapy with rebamipide and PPI had limited benefits compared with PPI monotherapy in the treatment of post-ESD gastric ulcer (UMIN Clinical Trials Registry, UMIN000007435).


Cost-Effective and Readily Replicable Surgical Simulation Model Improves Trainee Performance in Benchtop Robotic Urethrovesical Anastomosis.
Our objective was to develop a simple, cost-effective and reusable model for urethrovesical anastomosis for robotic-assisted radical prostatectomy and evaluate its impact on fundamental surgical skills and confidence of urology trainees. A model for the bladder, urethra and bony pelvis was created from materials easily purchased online. Each participant performed several trials of urethrovesical anastomosis using the da Vinci Si® surgical system. Pre-task confidence was assessed prior to each attempt. Two blinded researchers measured the following outcomes: time-to-anastomosis, number of suture throws, perpendicular needle entry and atraumatic needle driving. Integrity of the anastomosis was estimated by gravity filling and measuring pressure at which leakage occurred. These outcomes were translated into an independently validated Prostatectomy Assessment Competency Evaluation score. The model took 2 hours to create and total cost was 64 U.S. dollars. Twenty-one residents enrolled and demonstrated significant improvement in time-to-anastomosis, perpendicular needle driving, anastomotic pressure and total Prostatectomy Assessment Competency Evaluation score between the first and third trial. Pre-task confidence was measured on a Likert scale (1-5) and improved significantly over the 3 trials (Likert scale of 1.8, 2.8 and 3.3). We developed a cost-effective model of urethrovesical anastomosis that does not require the use of a 3D printer. This study demonstrates significant improvement of fundamental surgical skills and validated surgical assessment score for urology trainees over several trials. Our model shows potential for increasing accessibility of robotic training models for urological education. Additional investigation will be required to further assess the utility and validity of this model.


Lipoprotein catabolism in rats with portacaval anastomosis on various diets.
The catabolism of lipoproteins was measured in rats with a portacaval anastomosis and in intact control rats. Radioiodinated rat high density lipoproteins or human low density lipoproteins, the major cholesterol-bearing lipoproteins in rat or man respectively, were injected intravenously into rats. More than 90% of trace amounts of these lipoproteins were removed from plasma of rats with portacaval anastomosis and controls on standard chow at closely similar rates within 24 h. Also, [125I]high density lipoproteins left the plasma at comparable rates in controls and rats with portacaval anastomosis, whether fed with a cholesterol-free chow or a carbohydrate-rich lard chow. These dietary regimens were employed to avoid artifacts through a different development of the body weight in operated and control rats. A standard laboratory chow ad libitum led to weight loss in rats with portacaval anastomosis. Pair-fed with a cholesterol-free chow both groups of rats kept the same weight, but only with a carbohydrate-rich lard chow could the natural weight gain be achieved. In all rats with portacaval anastomosis liver weights were reduced and serum cholesterol decreased by 21-31% with the major change in high density lipoproteins. The findings suggest that cholesterol concentrations are not likely to be lowered in rats with portacaval anastomosis by enhanced lipoprotein catabolism.


The effect of two weeks of spinal manipulative therapy and home stretching exercises on pain and disability in patients with persistent or recurrent neck pain; a randomized controlled trial.
Recurrent or persistent neck pain affects a vast number of people globally, leading to reduced quality of life and high societal costs. Clinically, it is a difficult condition to manage, and treatment effect sizes are often moderate at best. Activity and manual therapy are first-line treatment options in current guidelines. We aimed to investigate the combination of home stretching exercises and spinal manipulative therapy in a multicentre randomized controlled clinical trial, carried out in multidiscipline ary primary care clinics. The treatment modalities utilized were spinal manipulative therapy and home stretching exercises compared to home stretching exercises alone. Both groups received 4 treatments for 2 weeks. The primary outcome was pain, where the subjective pain experience was investigated by assessing pain intensity (NRS - 11) and the quality of pain (McGill Pain Questionnaire). Neck disability and health status were secondary outcomes, measured using the Neck Disability Indexthe EQ-5D, respectively. One hundred thirty-one adult subjects were randomized to one of the two treatment groups. All subjects had experienced persistent or recurrent neck pain the previous 6 months and were blinded to the other group intervention. The clinicians provided treatment for subjects in both group and could not be blinded. The researchers collecting data were blinded to treatment allocation, as was the statistician performing data analyses. An intention-to-treat analysis was used. Sixty-six subjects were randomized to the intervention group, and sixty-five to the control group. For NRS - 11, a B-coefficient of - 0,01 was seen, indication a 0,01 improvement for the intervention group in relation to the control group at each time point with a p-value of 0,305. There were no statistically significant differences between groups for any of the outcome measures. Based on the current findings, there is no additional treatment effect from adding spinal manipulative therapy to neck stretching exercises over 2 weeks for patients with persistent or recurrent neck pain. The trial was registered 03/07/2018 at ClinicalTrials.gov , registration number: NCT03576846.


Association of oral bacteria with oral hygiene habits and self-reported gingival bleeding.
To describe associations of gingival bacterial composition and diversity with self-reported gingival bleeding and oral hygiene habits in a Norwegian regional-based population. We examined the microbiome composition of the gingival fluid (16S amplicon sequencing) in 484 adult participants (47% females; median age 28 years) in the Respiratory Health in Northern Europe, Spain and Australia (RHINESSA) study in Bergen, Norway. We explored bacterial diversity and abundance differences by the community periodontal index score, self-reported frequency of gingival bleeding, and oral hygiene habits. Gingival bacterial diversity increased with increasing frequency of self-reported gingival bleeding, with higher Shannon diversity index for "always" β = 0.51 and "often" β = 0.75 (p < .001) compared to "never" gingival bleeding. Frequent gingival bleeding was associated with higher abundance of several bacteria such as Porphyromonas endodontalis, Treponema denticola, and Fretibacterium spp., but lower abundance of bacteria within the gram-positive phyla Firmicutes and Actinobacteria. Flossing and rinsing with mouthwash twice daily were associated with higher total abundance of bacteria in the Proteobacteria phylum but with lower bacterial diversity compared to those who never flossed or never used mouthwash. A high frequency of self-reported gingival bleeding was associated with higher bacterial diversity than found in participants reporting no gingival bleeding and with higher total abundance of known periodontal pathogens such as Porphyromonas spp., Treponema spp., and Bacteroides spp.


Identification and expression profile of microRNA in seven tissues of the Golden snub-nosed monkey (Rhinopithecus roxellanae).
MicroRNAs (miRNAs) are key in the post-transcriptional regulation of gene expression and thus characterization of miRNAs and investigation of the relative abundance and specificity of tissue expression are essential for understanding gene expression in the golden snub-nosed monkey (GSM, Rhinopithecus roxellanae). Here, we report the first dataset of GSM miRNAs where we identified 460 miRNAs in seven tissues, with 246 conserved known mature miRNAs and 214 novel mature miRNAs. We determined miRNA abundance and expression in the seven tissues using a Tissue Specificity Index score and found that most novel GSM miRNAs showed a highly tissue-specific expression pattern. In particular, 67 novel miRNAs and the miR-34 family were expressed in abundance only in the lung. Five known miRNAs were highly abundant in digestive organs such as the pancreas and liver, and four novel miRNAs were highly expressed in the heart and muscle. Annotation of target genes of GSM miRNAs indicated that target genes were enriched in many important pathways, such as the HIF-1 signaling pathway and xenobiotic biodegradation-related pathways. Collectively, these results emphasize that miRNAs play important roles in GSM diet and high-elevation adaptation regulation. In summary, this study provides essential information on GSM miRNAs and will benefit further investigations of the function and mechanism of miRNAs in controlling gene expression in the GSM.


Establishing apoptosis resistant cell lines for improving protein productivity of cell culture.
The authors established apoptosis resistant COS-1, myeloma, hybridoma, and Friend leukemia cell lines by genetically engineering cells, aiming at more efficient protein production by cell culture. COS-1 cells, which are most widely used for eukariotic gene expression, were transfected with human bcl-2 gene. Both bcl-2 and mock transfected COS-1 cells were cultured at low (0.2%) serum concentration for 9 days. The final viable cell number of the bcl-2 transfected cells was ninefold of that of the mock transfectants. Both bcl-2 and mock transfectants were further transfected with the vector pcDNA-λ containing SV40 ori and immunoglobulin λ gene for transiently expressing λ protein. The bcl-2 expressing COS-1 cells produced more λ protein than the mock transfected COS-1 cells after 4 days posttransfection.Mouse myeloma p3-X63-Ag.8.653 cells, which are widely used as the partner for preparing hybridoma, and hybridoma 2E3 cells were transfected with human bcl-2 gene. Both bcl-2 transfected myeloma and hybridoma survived longer than the corresponding original cells in batch culture. The bcl-2 transfected 2E3 cells survived 2 to 4 four days longer in culture, producing 1.5- to 4-fold amount of antibody in comparison with the mock transfectants.Coexpression of bag-1 with bcl-2 improved survival of hybridoma 2E3 cells more than bcl-2 expression alone. The bag-1 and bcl-2 coexpressing cells produced more IgG than the the cells expressing bcl-2 alone.Apoptosis of Friend murine erythroleukemia(F-MEL) cells was suppressed with antisense c-jun expression. The antisense c-jun expressing cells survived 16 days at non-growth state.


[Delayed management of anorectal malformation with rectouretrobulbar fistula: a case study].
Ano-rectale malformations (ARM) are a spectrum of heterogeneous abnormalities in the development of the rectal canal. Its incidence is usually low and it is a little higher in some developing countries. Boys are more affected than girls and rectobulbar fistula associated with atresia of the anal canal is the most frequent disorder among them. We report the case of a 10-months old infant of male sex, whose mother lived in a mining area and had been complaining of fecaluria since the birth of her child. Physical examination showed fingerprint 1 cm below the intersection of the median raphe and the bi-ischiatic line. Paraclinical examinations showed no other associated malformation. Patient's management was based on anorectoplasty through abdominal and perineal approach with lower abdominoperineal reconstruction. In the post-operative period, the patient received antibiotic treatment and intravenous analgesia as well as dilations, which continued after discharge 2 weeks after surgery. No complications were observed and outcome was favorable.


Mining continuous intracranial EEG in focal canine epilepsy: Relating interictal bursts to seizure onsets.
Brain regions are localized for resection during epilepsy surgery based on rare seizures observed during a short period of intracranial electroencephalography (iEEG) monitoring. Interictal epileptiform bursts, which are more prevalent than seizures, may provide complementary information to aid in epilepsy evaluation. In this study, we leverage a long-term iEEG dataset from canines with naturally occurring epilepsy to investigate interictal bursts and their electrographic relationship to seizures. Four dogs were included in this study, each monitored previously with continuous iEEG for periods of 475.7, 329.9, 45.8, and 451.8 days, respectively, for a total of >11,000 h. Seizures and bursts were detected and validated by two board-certified epileptologists. A published Bayesian model was applied to analyze the dynamics of interictal epileptic bursts on EEG and compare them to seizures. In three dogs, bursts were stereotyped and found to be statistically similar to periods before or near seizure onsets. Seizures from one dog during status epilepticus were markedly different from other seizures in terms of burst similarity. Shorter epileptic bursts explored in this work have the potential to yield significant information about the distribution of epileptic events. In our data, bursts are at least an order of magnitude more prevalent than seizures and occur much more regularly. Our finding that bursts often display pronounced similarity to seizure onsets suggests that they contain relevant information about the epileptic networks from which they arise and may aide in the clinical evaluation of epilepsy in patients.


Mitochondrial Peptide Humanin Facilitates Chemoresistance in Glioblastoma Cells.
Humanin (HN) is a mitochondrial-derived peptide with robust cytoprotective effects in many cell types. Although the administration of HN analogs has been proposed to treat degenerative diseases, its role in the pathogenesis of cancer is poorly understood. Here, we evaluated whether HN affects the chemosensitivity of glioblastoma (GBM) cells. We found that chemotherapy upregulated HN expression in GBM cell lines and primary cultures derived from GBM biopsies. An HN analog (HNGF6A) boosted chemoresistance, increased the migration of GBM cells and improved their capacity to induce endothelial cell migration and proliferation. Chemotherapy also upregulated FPR2 expression, an HN membrane-bound receptor, and the HNGF6A cytoprotective effects were inhibited by an FPR2 receptor antagonist (WRW4). These effects were observed in glioma cells with heterogeneous genetic backgrounds, i.e., glioma cells with wild-type (wtIDH) and mutated (mIDH) isocitrate dehydrogenase. HN silencing using a baculoviral vector that encodes for a specific shRNA for HN (BV.shHN) reduced chemoresistance, and impaired the migration and proangiogenic capacity of GBM cells. Taken together, our findings suggest that HN boosts the hallmark characteristics of GBM, i.e., chemoresistance, migration and endothelial cell proliferation. Thus, strategies that inhibit the HN/FPR2 pathway may improve the response of GBM to standard therapy.


Identification of proteins from formalin-fixed paraffin-embedded cells by LC-MS/MS.
There exists a need for robust approaches for tandem mass spectrometry (MS/MS)-based identification of proteins in formalin-fixed paraffin-embedded (FFPE) material. We demonstrate herein the identification of proteins in FFPE material using enzymatic cleavage for extraction of peptides from the FFPE specimen and liquid chromatography (LC) followed by MS/MS. We identified 324 proteins from a 3-year-old FFPE cell-block of a human lymphoma cell line. The identified proteins were assigned to the membrane, cytosol and nucleus, with diverse cellular functions. The results were comparable to those obtained with lysates from a fresh specimen of the lymphoma cell line. Western blotting analysis and immunofluorescence microscopy confirmed the expression of selected proteins. The functional significance of one protein (PKC eta) was validated using a PKC inhibitory peptide which resulted in lymphoma cell death in vitro. The ability to identify proteins from FFPE specimens has significant implications for MS/MS-based proteomics of vast repositories of archival primary tissue samples for disease-related discovery research.


How Safe Is the Use of Intrathecal Vancomycin?
Central nervous system infection after neurosurgical procedures is a severe complication with high morbidity rates and sometimes mortality. Our experimental study aimed to investigate the biochemical and histopathologic effects of vancomycin on neural tissues when applied to the cisterna magna. Wistar albino rats were randomly divided into 4 groups: Control (Group 1) and different vancomycin dose groups (Groups 2, 3, and 4). In Group 1, 0.1 mL cerebrospinal fluid was drained from the cisterna magna and 0.1 mL 0.9% NaCI (normal saline) was administered into the subarachnoid space. In the study groups, 0.1 mL cerebrospinal fluid was drained from the cisterna magna and 0.1 mg/200g rat per day (Group 2), 0.2 mg/200g rat per day (Group 3), and 0.4 mg/200g rat per day (Group 4) vancomycin were administered into the subarachnoid space for 7 days. All rats were sacrificed on the eighth day. Serum superoxide dismutase and catalase levels were measured. Histopathologic and immunohistochemical analyses were conducted. The findings showed that the administration of 0.2 and 0.4 mg/kg doses had significant differences in superoxide dismutase and catalase activity compared with the controls (P < 0.05). These vancomycin doses also induced the apoptotic process, and the enzyme activity results correlated with immunohistochemical results. Dose-related neurotoxicity of intrathecal vancomycin was shown at the cellular level. The importance of dose regulation of intrathecal vancomycin has come into view. To our knowledge, this is the first study in the literature that has investigated the neurotoxic effects of vancomycin.


Non-steroidal anti-inflammatories--XII. Mode of action of anti-inflammatory methane sulfonanilides.
The influence of anti-inflammatory methane sulfonanilides on the arachidonic acid metabolism in ram seminal vesicle homogenate depends on the pK3 of the individual substance: more acidic compounds suppress, less acidic compounds stimulate PG synthesis and enhance the PGH2/PGG2 ratio. As oxygen radicals are liberated during the conversion of PGG2 to PGH2 it is suggested that the less acidic methane sulfonanilides are scavengers of the (pro-inflammatory) oxygen radicals.


The 100 top-cited articles in orthodontics from 1975 to 2011.
To identify the 100 top-cited articles published in orthodontics journals and to analyze their characteristics to investigate the achievement and development of orthodontics research in past decades. The Institute for Scientific Information Web of Knowledge Database and the 2011 Journal Citation Report Science Editions were used to retrieve the 100 top-cited articles published in orthodontics journals since 1975. Some basic information was collected by the Analyze Tool on the Web of Science, including citation time, publication title, journal name, publication year, and country and institution of origin. A further study was then performed to determine authorship, article type, field of study, study design, and level of evidence. The 100 target articles were retrieved from three journals: American Journal of Orthodontics and Dentofacial Orthopedics (n = 74), The Angle Orthodontist (n = 15), and European Journal of Orthodontics (n = 11). Since 1975, the articles cited 89 to 545 times mainly originated from the United States, and the overwhelming majority of articles were clinical. The most common study design was case series; 40 articles were classified as level IV and 12 as level V evidence. The 100 top-cited articles in orthodontics are generally old articles, rarely possessing high-level evidence.


Cytokinesis depends on the motor domains of myosin-II in fission yeast but not in budding yeast.
Budding yeast possesses one myosin-II, Myo1p, whereas fission yeast has two, Myo2p and Myp2p, all of which contribute to cytokinesis. We find that chimeras consisting of Myo2p or Myp2p motor domains fused to the tail of Myo1p are fully functional in supporting budding yeast cytokinesis. Remarkably, the tail alone of budding yeast Myo1p localizes to the contractile ring, supporting both its constriction and cytokinesis. In contrast, fission yeast Myo2p and Myp2p require both the catalytic head domain as well as tail domains for function, with the tails providing distinct functions (Bezanilla and Pollard, 2000). Myo1p is the first example of a myosin whose cellular function does not require a catalytic motor domain revealing a novel mechanism of action for budding yeast myosin-II independent of actin binding and ATPase activity.


We need to rename 'whistleblowing' if we really want people to speak up.
The recently published NHS improvement guide, Freedom to Speak Up, has many features to commend it.


Anticancer Opportunity Created by Loss of Tumor Suppressor Genes.
Deletion of oncosuppressors occurs frequently in the cancer genome. A great deal of effort has been made to therapeutically restore the lost function of oncosuppressors, with little clinically translatable success, however. Reassuringly, besides the disappointing restoration endeavors, oncosuppressor loss can be therapeutically exploited in several other ways, such as the "synthetic lethality" strategies and the "therapeutic vulnerability" created by codeletion of neighboring genes. The study by Liu et al showed that codeletion of p53 and a neighboring essential gene POLR2A rendered colon cancer cells highly sensitive to further inhibition of POLR2A both in vitro and in vivo In recent years, several studies have reported similar phenomenon in a wide range of cancer types. In this focus article, we will introduce several kinds of anticancer opportunities created by the loss of oncosuppressors and discuss their mechanisms. Given the frequency of oncosuppressor loss in cancer, its therapeutic exploitation rather merits further investigation and may open a new window for oncotherapy.


Anticoagulation for atrial fibrillation.
For over two decades, valuable insights have been accumulated from epidemiologic studies and randomized trials about the risks for and prevention of AF-related stroke. AF substantially raises the risk of stroke, most likely through an atrio-embolic mechanism. Warfarin and other members of its class of oral anticoagulants targeted at an INR of 2.5 can abrogate the risk of stroke attributable to AF effectively and fairly safely. High-quality management of anticoagulation can be achieved in usual clinical care. These insights have important implications for the care of individual patients and more generally for public health. Future research is needed to specify the risk of stroke and hemorrhage among patients with AF better, particularly among older individuals, to optimize use of antithrombotic agents, and to define the role of recently developed antithrombotic drugs and invasive nondrug approaches.


Non-Hodgkin's lymphoma as a cause of hypoadrenalism.
Non-Hodgkin's lymphoma which presents as adrenocortical insufficiency is rare, however, it should be a well-recognised entity because, in addition to specific chemotherapy or radiotherapy, it needs hormonal replacement. We report a patient with diffuse histiocytic lymphoma, who had remission for seven years and relapsed with Addisonian crisis.


Early detection and stratification of lung cancer aided by a cost-effective assay targeting circulating tumor DNA (ctDNA) methylation.
Detection of lung cancer at earlier stage can greatly improve patient survival. We aim to develop, validate, and implement a cost-effective ctDNA-methylation-based plasma test to aid lung cancer early detection. Case-control studies were designed to select the most relevant markers to lung cancer. Patients with lung cancer or benign lung disease and healthy individuals were recruited from different clinical centers. A multi-locus qPCR assay, LunaCAM, was developed for lung cancer alertness by ctDNA methylation. Two LunaCAM models were built for screening (-S) or diagnostic aid (-D) to favor sensitivity or specificity, respectively. The performance of the models was validated for different intended uses in clinics. Profiling DNA methylation on 429 plasma samples including 209 lung cancer, 123 benign diseases and 97 healthy participants identified the top markers that detected lung cancer from benign diseases and healthy with an AUC of 0.85 and 0.95, respectively. The most effective methylation markers were verified individually in 40 tissues and 169 plasma samples to develop LunaCAM assay. Two models corresponding to different intended uses were trained with 513 plasma samples, and validated with an independent collection of 172 plasma samples. In validation, LunaCAM-S model achieved an AUC of 0.90 (95% CI: 0.88-0.94) between lung cancer and healthy individuals, whereas LunaCAM-D model stratified lung cancer from benign pulmonary diseases with an AUC of 0.81 (95% CI: 0.78-0.86). When implemented sequentially in the validation set, LunaCAM-S enables to identify 58 patients of lung cancer (90.6% sensitivity), followed by LunaCAM-D to remove 20 patients with no evidence of cancer (83.3% specificity). LunaCAM-D significantly outperformed the blood test of carcinoembryonic antigen (CEA), and the combined model can further improve the predictive power for lung cancer to an overall AUC of 0.86. We developed two different models by ctDNA methylation assay to sensitively detect early-stage lung cancer or specifically classify lung benign diseases. Implemented at different clinical settings, LunaCAM models has a potential to provide a facile and inexpensive avenue for early screening and diagnostic aids for lung cancer.


Pediatric Program Directors Should Have an Active Presence on Twitter.
For academic pediatricians, social media has become an important avenue for professional development through continuing education, professional networking, and academic collaboration. Pediatric residency program directors have recognized additional benefits of social media engagement via program promotion and resident recruitment. The novel coronavirus disease 2019 (COVID-19) pandemic and subsequent move to virtual interviews for the 2020-2021 residency interview season have created a new urgency for pediatric program directors to establish an active social media presence, primarily as a means to engage applicants and provide them with information in lieu of cancelled away rotations and in-person interviews. Twitter is a free microblogging and social networking platform that allows real-time engagement among academic pediatricians. Here, we make the case that all pediatric program directors should have an active presence on Twitter.


West syndrome with periventricular leukomalacia: a morphometric MRI study.
A morphometric magnetic resonance imaging study was performed, and the results were compared among three groups (group 1, periventricular leukomalacia patients with West syndrome; group 2, periventricular leukomalacia patients without West syndrome; and group 3, control patients) to clarify the characteristics and cause of West syndrome. This study included 21 infants (11 males and 10 females, 7 months to 2 years 8 months old) born at 24-32 weeks of gestation and weighing 625-1,908 gm. The Evans ratio, ratio of the posterior horns, Cella media index, width of the third ventricle, and the areas of the midbrain, pons, and medulla oblongata were measured and compared among the three groups. There were no differences of gestation or birth weight among the three groups. The Evans ratio, ratio of the posterior horns, Cella media index, and width of the third ventricle were larger in group 1 than in groups 2 and 3. The ratio of the posterior horns and Cella media index were larger in group 2 than in group 3, although the width of the third ventricle was not. Myelination was delayed in all patients in group 1 and in two patients in group 2. In group 1 the areas of the midbrain and pons were smaller than in groups 2 and 3 and the medulla oblongata was smaller than in group 3, although there were no differences in midbrain, pons, and medulla oblongata between groups 2 and 3. Although the infants with periventricular leukomalacia and West syndrome frequently demonstrated marked ventricular dilatation and delayed myelination, the atrophy of midbrain and pons was the most characteristic, and the damage may cause West syndrome.


Anthropogenic and natural hydrocarbons in water and sediments of the Kara Sea.
The results on the content and composition of aliphatic (AHСs) and polycyclic aromatic hydrocarbons (PAHs) at the geochemical barrier between the Kara Sea and the Ob and Yenisei Rivers are discussed in comparison with those in the mouth areas of the Lena, Khatanga, Indigirka and Kolyma. It has been established that hydrocarbon concentrations sharply decrease in the river-sea mixing zones, and their composition, and the ratio of their dissolved and suspended forms change. Besides, the research season plays a great role, as the highest content of hydrocarbons and particulate matter was found in the Ob Gulf during the flood, when the concentrations of AHCs in surface waters reached 325 μg/L, PAHs -15 ng/L, particulate matter-15.5 mg/L. In the Kara Sea itself, AHC concentrations increased on average from 20 to 59 μg/L, that is most likely associated with the melting of seasonal ice and floods. Anthropogenic influence was found in river waters, where the proportion of naphthalenes, which marks oil products, and pyrene, which marks pyrogenic PAHs, is increased. In the Ob Gulf is linked to an increase in navigation along the Northern Sea Route and the functioning of the port of Sabetta. In the mouth area of the Yenisei, the concentrations of both AHC and PAH in surface waters and sediments varied in a smaller range than in the mouth area of the Ob, which is due to the nature of the rocks that make up the catchment areas of these rivers, as well as the regulation of the Yenisei runoff. The hydrological features of the estuarine areas can lead to increased AHC concentrations (the mouth of the Khatanga, up to 189 μg/L, 2.6 μg/mg SPM). The minimum AHC content in SPM and sediments in the river transects was established in the East Siberian rivers (Indigirka, Kolyma), associated with the decreasing productivity in waters from the Western Arctic seas to the Eastern.


Role of solid carriers in pharmaceutical performance of solid supersaturable SEDDS of celecoxib.
Self emulsifying drug delivery system (SEDDS) has been increasingly used for improving the oral bioavailability of poorly water soluble drugs. SEDDS can be solidified by adsorbing them on different solid carriers. In the present study, the impact of properties of solid carrier on drug release profile from solid SEDDS was investigated. Celecoxib (CEL) loaded supersaturable SEDDS (S-SEDDS) was prepared and optimized by using optimal response surface design. Optimum composition of S-SEDDS corresponded to 10:45:45% v/v ratio of oil (Capryol 90), surfactant (Tween 20) and cosurfactant (Transcutol HP) with Soluplus (40 mg) as precipitation inhibitor. Different grades of silicon dioxide were selected based on their properties like surface area, porosity and hydrophobicity-hydrophilicity, and used for preparation of solid S-SEDDS (SS-SEDDS) by adsorption method. All SS-SEDDS formulations in release studies, gave droplet size, PDI and zeta potential similar to S-SEDDS. The percent drug release after 120min from CEL powder, S-SEDDS and SS-SEDDS with Sylysia 350 fcp, Aerosil 300 Pharma, Aerosil 200 Pharma and Aerosil R 972 Pharma was found to be 0.58%, 100%, 38.44%, 9.63%, 2.53% and 5.99%, respectively. Drug release profiles were compared by using model independent methods. The differential drug release behavior of SS-SEDDS was attributed to the different physico-chemical properties of solid carriers. SS-SEDDS with Sylysia 350 fcp showed higher drug release and greater dissolution efficiency. Oral bioavailability study also demonstrated 2.34 fold increase in Cmax and 4.82 fold increase in AUC (0-24h) when compared with CEL powder. This study highlights the rational for selection of solid carriers in the formulation development of solid SEDDS.


Reduced Platelet Adhesion for Blended Electrospun Meshes with Low Amounts of Collagen Type I.
A clinically approved, tissue engineered graft is needed as an alternative for small-diameter artery replacement. Collagen type I is commonly investigated for naturally derived grafts. However, collagen promotes thrombosis, currently requiring a graft pre-seeding step. This study investigates unique impacts of blending low collagen amounts with synthetic polymers on scaffold platelet response, which would allow for viable acellular grafts that can endothelialize in vivo. While platelet adhesion and activation are confirmed to be high with 50% collagen samples, low collagen ratios surprisingly exhibit the opposite, anti-thrombogenic effect. Different platelet interactions in these blended materials can be related to collagen structure. Low collagen ratios show homogenous distribution of the components within individual fibers. Importantly, blended collagen scaffolds exhibit significant differences from gelatin scaffolds, including retaining percentage of collagen after incubation. These findings correlate with functional benefits including better endothelial cell spreading on collagen versus gelatin blended materials. This appears to differ from the current paradigm that processing with harsh solvents will irreversibly denature collagen into less desirable gelatin, but an important distinction is the interaction between collagen and synthetic materials during processing. Overall, excellent anti-thrombogenic properties of low collagen blends and benefits after grafting show promise for this vascular graft strategy.


Inactivation of Escherichia coli O157:H7, Salmonella typhimurium, and Listeria monocytogenes on cherry tomatoes and oranges by superheated steam.
This study was performed to compare the effectiveness of saturated steam and superheated steam for the inactivation of Escherichia coli O157:H7, Salmonella typhimurium, and Listeria monocytogenes on the surface of cherry tomatoes and oranges. It also determined the effect of the steam processes on the color, texture, Vitamin C content, and antioxidant capacity and changes in these parameters during chilled storage. Cherry tomatoes and oranges inoculated with the three foodborne pathogens were treated with saturated steam at 100 °C and superheated steam at 125, 150, 175, and 200 °C for various time intervals. After the cherry tomatoes and oranges were exposed to superheated steam at 200 °C for 3 or 20 s, all tested pathogens were reduced to below the detection limit (1 or 1.7 log, respectively) without significant changes in color, texture, vitamin C content, and antioxidant capacity (P > .05) at 4 °C for up to 9 days. Our results suggest that superheated steam treatment can be effective at decreasing pathogen populations when compared to saturated steam, without significant quality deterioration, and thus, this technique demonstrates great potential to improve the microbial safety of fresh produce.


The role of the left hippocampal region in the acquisition and retention of story content.
Thirteen normal control subjects and 62 patients who had undergone either a unilateral temporal or a unilateral frontal lobectomy learned the content of a short prose passage to a strict criterion. Compared to other subject, patients with left temporal-lobe excision took longer to learn the story content and, within this group, the slowest rate noted was for patients with extensive removal from the hippocampal region. When retention of the material was tested after a 20 min delay, only the group with large excisions from the left hippocampus and/or parahippocampal gyrus was impaired. This finding of abnormally rapid forgetting of material learned to criterion highlights the role of the left hippocampal region in the long-term maintenance of verbal information presented in a context.


Identification of de novo chromosomal markers and derivatives by spectral karyotyping.
Despite major advances in molecular cytogenetics during the past decade and the important diagnostic role that fluorescence in situ hybridization (FISH) plays in the characterization of chromosomal abnormalities, the usefulness of this technique remains limited by the number of spectrally distinguishable fluorochromes or fluorochrome combinations. Overcoming this major limitation would allow one to use FISH to screen the whole human genome for chromosomal aberrations which, until recently, was possible only through conventional karyotyping. A recently described molecular cytogenetics technology, 24-color FISH using spectral karyotyping (SKY), permits the simultaneous visualization of all human chromosomes in 24 different colors. Most chromosomal aberrations detected during cytogenetic evaluation can be resolved using routine cytogenetic techniques alone or in combination with single- or dual-color FISH. However, some cases remain unresolved, in particular de novo supernumerary marker chromosomes and de novo unbalanced structural rearrangements. These findings cause particular diagnostic and counseling concerns when detected during prenatal diagnosis. The purpose of this report is to demonstrate the application of SKY in the characterization of these de novo structural chromosomal abnormalities. Eight cases are described in this report. SKY has considerable diagnostic applications in prenatal diagnosis because of its reliability and speed. The identification of the chromosomal origin of markers and unbalanced translocations provides the patient, physician, and genetic counselor with better predictive information on the phenotype of the carrier.


Experimental research on surface acoustic wave microfluidic atomization for drug delivery.
This paper demonstrates that surface acoustic wave (SAW) atomization can produce suitable aerosol concentration and size distribution for efficient inhaled lung drug delivery and is a potential atomization device for asthma treatment. Using the SAW device, we present comprehensive experimental results exploring the complexity of the acoustic atomization process and the influence of input power, device frequency, and liquid flow rate on aerosol size distribution. It is hoped that these studies will explain the mechanism of SAW atomization aerosol generation and how they can be controlled. The insights from the high-speed flow visualization studies reveal that it is possible by setting the input power above 4.17 W, thus allowing atomization to occur from a relatively thin film, forming dense, monodisperse aerosols. Moreover, we found that the aerosol droplet size can be effectively changed by adjusting the input power and liquid flow rate to change the film conditions. In this work, we proposed a method to realize drug atomization by a microfluidic channel. A SU-8 flow channel was prepared on the surface of a piezoelectric substrate by photolithography technology. Combined with the silicon dioxide coating process and PDMS process closed microfluidic channel was prepared, and continuous drug atomization was provided to improve the deposition efficiency of drug atomization by microfluidic.


Ventilatory Support Use in Hospitalized Patients With Community-Acquired Pneumonia. Fifteen-year Trends in Spain (2001-2015).
We examined fifteen years trends (2001-2015) in the use of non-invasive ventilation (NIV), invasive mechanical ventilation (IMV) or both (NIV+IMV) among patients hospitalized for community acquired pneumonia (CAP). We also analyzed trends overtime and the influence of patient factors in the in-hospital mortality (IHM) after receiving NIV, IMV or NIV+IMV. Observational retrospective epidemiological study. Our data source was the Spanish National Hospital Discharge Database. Over a total of 1,486,240 hospitalized patients with CAP, we identified 56,158 who had received ventilator support in Spain over the study period. Of them, 54.82% received NIV, 37.04% IMV and 8.14% both procedures. The use of NIV and NIV+IMV increased significantly (p<0.001) over time (from 0.91 to 12.84 per 100.000 inhabitant and from 0.23 to 1.19 per 100.000 inhabitants, respectively), while the IMV utilization decreased (from 3.55 to 2.79 per 100,000 inhabitants; p<0.001). Patients receiving NIV were the oldest and had the highest mean value in the Charlson comorbidity index (CCI) score and readmission rate. Patients who received only IMV had the highest IHM. Factors associated with IHM for all groups analyzed included age, comorbidities and readmission. IHM decreased significantly over time in patients with CAP who received NIV, IMV and NIV+IMV. We found an increase in NIV use and a decline in IMV utilization in patients hospitalized for CAP over the study period. Patients receiving NIV were the oldest and had the highest CCI score and readmission rate. IHM decreased significantly over time in patients with CAP who received NIV, IMV and NIV+IMV.


Magneto-optical visualization of metal-loss defects in a ferromagnetic plate: experimental verification of theoretical modeling.
Rare-earth iron garnet films with in-plane magnetic anisotropy grown on [111]-oriented substrates are promising for the visualization of magnetic leakage fields in nondestructive evaluation. Such magneto-optical films have to be specifically engineered, and we give an example of this technology. To assess the validity and accuracy of finite-element calculations of a magnetization assembly combined with the physical modeling of the image formation, comparisons between calculated and experimentally obtained magneto-optical images of metal-loss defects have been made. A convincing quantitative agreement is demonstrated. It is shown that both physical and computer modeling techniques allow for a predictive engineering design of the prospective applications and provide greater insight into the method.


Measurement of DNA breakage in spermiogenic germ-cell stages of mice exposed to ethylene oxide, using an alkaline elution procedure.
DNA breakage in spermiogenic stages of the mouse was studied after exposure to ethylene oxide (EtO), using an alkaline elution technique. At daily intervals over a 23-day period following i.p. injection of 100 mg EtO/kg, mature spermatozoa were recovered from treated ([3H]dThd-labeled) and control ([14C]dThd-labeled) animals, lysed together on polycarbonate filters, and the DNA was eluted with a high pH (12.2) buffer. Elution of germ-cell DNA from EtO-exposed animals increased (more DNA strand breaks) in stages sensitive to the genetic effects of EtO (late spermatids to early spermatozoa). The stage-related pattern of EtO-induced DNA breakage paralleled the pattern of sperm alkylation and protamine alkylation found to be produced by EtO in an earlier study (Sega and Owens, 1987). At 9 days posttreatment (sperm sampled were in late-spermatid stages at the time of EtO exposure) the amount of sperm DNA eluted did not change significantly over a pH range of 11.6-12.8, indicating that, at the time of assay, DNA breaks were already present in the sperm.


Dietary hydrolyzed wheat gluten supplementation ameliorated intestinal barrier dysfunctions of broilers challenged with Escherichia coli O78.
This study aimed to investigate a protective effect of hydrolyzed wheat gluten (HWG) on Escherichia coli (E. coli)-induced intestinal barrier dysfunctions in broilers. Broilers fed a basal diet unsupplemented or supplemented with HWG (0.5% or 1%) were intraperitoneally injected with either E. coli O78 suspension (108 CFU/mL) or equal volume of vehicle on d 18 of age. Blood and tissue samples were collected 3rd d post infection. The results showed that E. coli-infection increased immune-organ indexes of spleen and thymus, enhanced serum diamine oxidase (DAO) level, impaired ileal villus structure and reduced tight junction mRNA levels (Occludin, Claudin-1, ZO-1, P < 0.05), while increased mRNA levels of inflammatory cytokines (TNF-α, IFN-γ, IL-1β, IL-6, IL-8) and TLR4 in the ileum of broilers (P < 0.05). The effects of E. coli O78 challenge on organ indexes of spleen and thymus, serum DAO level, mRNA levels of tight junctions were alleviated by 1% HWG supplementation, the upregulations of IL-1β and TLR4 were prevented by 0.5% HWG supplementation (P < 0.05). In addition, increased IFN-γ of E. coli-infected broilers was abrogated by 0.5% or 1% HWG supplementation (P < 0.05). In summary, dietary HWG supplementation ameliorated intestinal barrier dysfunctions triggered by E. coli-infection in the ileum of broilers. HWG supplementation might be a nutritional strategy to improve the intestinal mucosal barrier function of broilers.


The protective roles of nitric oxide and superoxide dismutase in adriamycin-induced cardiotoxicity.
Treatment with adriamycin (ADR) is associated with cardiotoxicity mediated through the generation of superoxide (O2*-). Because nitric oxide (*NO) reacts with O2*-, generating peroxynitrite, we hypothesized that decreased *NO production would lead to protection in acute cardiac injury. We investigated the role of decreased *NO levels in exacerbation of ADR-induced cardiotoxicity in vivo using iNOS (-/-) mice. Pathology, biochemical injury markers, and cardiac function were used to assess ADR-induced cardiac injury. Ultrastructural analysis demonstrated that iNOS (-/-) mice exhibited extensive cytoplasmic swelling and degeneration of mitochondria when compared to wildtype mice following treatment with ADR. Mice lacking iNOS exhibited a decrease in resting indices of cardiac function as well as an impairment in the positive inotropic actions of isoproterenol following treatment with ADR compared to nTg mice. Cardiac troponin, creatine phosphokinase, and lactate dehydrogenase levels were significantly increased after treatment in iNOS (-/-) mice as compared to controls and wildtype mice. These results indicate that a lack of *NO production by iNOS caused significantly enhanced cardiac injury. However, when iNOS (-/-) mice were crossed with manganese superoxide dismutase (MnSOD)-overexpressing animals, mitochondrial injury was ameliorated to the level of the wild type. These findings suggest that reduction of *NO levels mediated by ADR treatment leads to increased cardiac mitochondrial injury that can be attenuated by a compensatory increase in MnSOD.


Seminars in Thoracic and Cardiovascular Surgery - Current Readings: Adult Congenital Heart Surgery.
While the incidence of congenital heart disease has remained relatively stable, survival has increased dramatically. Many of these patients require lifelong specialized cardiac care to address sequelae and residual disease. In order to best meet the increasing demands and needs of this heterogeneous group of survivors, multidisciplinary specialty training, lesion specific outcomes research and both personnel and financial resources must be dedicated to their care.


Synthesis and mesomorphic behavior of a donor-acceptor-type hexaazatriphenylene.
[structure: see text] A new donor-acceptor, 1,4,5,8,9,12-hexaazatriphenylene HATCNOR(n), is described. The synthesis of HATCNOR1 and HATCNOR6 is achieved by the regioselective displacement of 1,4,5,8,9,12-hexaazatriphenylene hexacarbonitrile (HATCN) with an alkoxy group. The X-ray analysis revealed self-assembly of HATCNOR1 in the solid state. HATCNOR6 is the new difunctionalized hexaazatriphenylene discotic liquid crystal.


Mechanisms by which the thiazolidinedione troglitazone protects against sucrose-induced hepatic fat accumulation and hyperinsulinaemia.
Thiazolidinediones (TZD) are known to ameliorate fatty liver in type 2 diabetes. To date, the underlying mechanisms of their hepatic actions remain unclear. Hepatic triglyceride content and export rates were assessed in 2 week high-sucrose-fed Wistar rats treated with troglitazone and compared with untreated high-sucrose rodent controls. Fractional de novo lipogenesis (DNL) contributions to hepatic triglyceride were quantified by analysis of triglyceride enrichment from deuterated water. Hepatic insulin clearance and NO status during a meal tolerance test were also evaluated. TZD significantly reduced hepatic triglyceride (P < 0.01) by 48%, decreased DNL contribution to hepatic triglyceride (P < 0.01) and increased postprandial non-esterified fatty acids clearance rates (P < 0.01) in comparison with the high-sucrose rodent control group. During a meal tolerance test, plasma insulin AUC was significantly lower (P < 0.01), while blood glucose and plasma C-peptide levels were not different. Insulin clearance was increased (P < 0.001) by 24% and was associated with a 22% augmentation of hepatic insulin-degrading enzyme activity (P < 0.05). Finally, hepatic NO was decreased by 24% (P < 0.05). Overall, TZD show direct actions on liver by reducing hepatic DNL and increasing hepatic insulin clearance. The alterations in hepatic insulin clearance were associated with changes in insulin-degrading enzyme activity, with possible modulation of NO levels.


Nano-Surveillance: Tracking Individual Molecules in a Sea of Chromatin.
Chromatin is the epigenomic platform for diverse nuclear processes such as DNA repair, replication, transcription, telomere, and centromere function. In cancer cells, mutations in key processes result in DNA amplification, chromosome translocations, and chromothripsis, severely distorting the natural chromatin state. In normal and diseased states, dozens of chromatin effectors alter the physical integrity and dynamics of chromatin at the level of both single nucleosomes and arrays of nucleosomes folded into 3-dimensional shapes. Integrating these length scales, from the 10 nm sized nucleosome to mitotic chromosomes, whilst jostling within the crowded environment of the cell, cannot yet be achieved by a single technology. In this review, we discuss tools that have proven powerful in the investigation of nucleosome and chromatin fiber dynamics. We also provide a deeper focus into atomic force microscopy (AFM) applications that can bridge diverse length and time scales. Using time course AFM, we observe that chromatin condensation by H1.5 is dynamic, whereas using nano-indentation force spectroscopy we observe that both histone variants and nucleosome binding partners alter material properties of individual nucleosomes. Finally, we demonstrate how high-speed AFM can visualize plasmid DNA dynamics, intermittent nucleosome-nucleosome contacts, and changes in nucleosome phasing along a contiguous chromatin fiber. Altogether, the development of innovative technologies holds the promise of revealing the secret lives of nucleosomes, potentially bridging the gaps in our understanding of how chromatin works within living cells and tissues.


A review of movement disorders in persons living with HIV.
The human immunodeficiency virus (HIV) causes movement disorders in persons living with HIV (PLH). We conducted a systematic review on the spectrum of movement disorders in PLH using standard terms for each of the phenomenologies and HIV. Movement disorders in PLH were commonly attributed to opportunistic infections (OI), dopamine receptor blockade reactions, HIV-associated dementia (HAD), presented during seroconversion, developed due to drug reactions or antiretroviral therapy (ART) itself and lastly, movement disorders occurred as a consequence of the HIV-virus. Parkinsonism in ART naïve PLH was associated with shorter survival, however when Parkinsonism presented in PLH on ART, the syndrome was indistinguishable from Idiopathic Parkinson's disease and responded to therapy. Tremor was often postural due to HAD, drugs or OI. Generalized chorea was most frequent in HIV encephalopathy and toxoplasmosis gondii caused most cases of hemichorea. Ataxia was strongly associated with JCV infection, ART efavirenz toxicity or due to HIV itself. Dystonia was reported in HAD, secondary to drugs and atypical facial dystonias. Both cortical/subcortical and segmental/spinal origin myoclonus were noted mainly associated with HAD. In patients with HIV related opsoclonus-myoclonus-ataxia-syndrome, seroconversion illness was the commonest cause of followed by IRIS and CSF HIV viral escape phenomenon. Aetiology of movement disorders in PLH depend on the treatment state. Untreated, PLH are prone to develop OI and HAD and movement disorders. However, as the number of PLH on ART increase and survive longer, the frequency of ART and non-AIDS related complications are likely to increase.


The emergence of autoclitic frames in atypically and typically developing children as a function of multiple exemplar instruction.
In two experiments, we tested the effect of multiple exemplar instruction (MEI) for training sets on the emergence of autoclitic frames for spatial relations for novel tacts and mands. In Experiment 1, we used a replicated pre- and post-intervention probe design with four students with significant learning disabilities to test for acquisition of four autoclitic frames with novel tacts and mands before and after MEI. The untaught topographies emerged for all participants. In Experiment 2, we used a multiple probe design to test the effects of the MEI procedures on the same responses in four typically developing, bilingual students. The novel usage emerged for all participants. In the latter experiment, the children demonstrated untaught usage of mand or tact frames regardless of whether they were taught to respond in either listener or speaker functions alone or across listener and speaker functions. The findings are discussed in terms of the role of MEI in the formation of abstractions.


Adoptive Transfer of Cryptococcus neoformans-Specific CD4 T-Cells to Study Anti-fungal Lymphocyte Responses In Vivo.
CD4 T-cells are important for long-term control and clearance of several fungal infections in humans, particularly those caused by Cryptococcus species. Understanding the mechanisms underlying protective T-cell immunity against fungal infection is critical for developing mechanistic insights into the pathogenesis of the disease. Here, we describe a protocol that enables analysis of fungal-specific CD4 T-cell responses in vivo, using adoptive transfer of fungal-specific T-cell receptor (TCR) transgenic CD4 T-cells. While the protocol here uses a TCR transgenic model reactive to peptide deriving from Cryptococcus neoformans, this method could be adapted to other fungal infection experimental settings.


Eomes restricts Brachyury functions at the onset of mouse gastrulation.
Mammalian specification of mesoderm and definitive endoderm (DE) is instructed by the two related Tbx transcription factors (TFs) Eomesodermin (Eomes) and Brachyury sharing partially redundant functions. Gross differences in mutant embryonic phenotypes suggest specific functions of each TF. To date, the molecular details of separated lineage-specific gene regulation by Eomes and Brachyury remain poorly understood. Here, we combine mouse embryonic and stem-cell-based analyses to delineate the non-overlapping, lineage-specific transcriptional activities. On a genome-wide scale, binding of both TFs overlaps at promoters of target genes but shows specificity for distal enhancer regions that is conferred by differences in Tbx DNA-binding motifs. The unique binding to enhancer sites instructs the specification of anterior mesoderm (AM) and DE by Eomes and caudal mesoderm by Brachyury. Remarkably, EOMES antagonizes BRACHYURY gene regulatory functions in coexpressing cells during early gastrulation to ensure the proper sequence of early AM and DE lineage specification followed by posterior mesoderm derivatives.


A synthetic enzymatic pathway for extremely thermophilic acetone production based on the unexpectedly thermostable acetoacetate decarboxylase from Clostridium acetobutylicum.
One potential advantage of an extremely thermophilic metabolic engineering host (T opt ≥ 70°C) is facilitated recovery of volatile chemicals from the vapor phase of an active fermenting culture. This process would reduce purification costs and concomitantly alleviate toxicity to the cells by continuously removing solvent fermentation products such as acetone or ethanol, a process we are calling "bio-reactive distillation." Although extremely thermophilic heterologous metabolic pathways can be inspired by existing mesophilic versions, they require thermostable homologs of the constituent enzymes if they are to be utilized in extremely thermophilic bacteria or archaea. Production of acetone from acetyl-CoA and acetate in the mesophilic bacterium Clostridium acetobutylicum utilizes three enzymes: thiolase, acetoacetyl-CoA: acetate CoA transferase (CtfAB), and acetoacetate decarboxylase (Adc). Previously reported biocatalytic pathways for acetone production were demonstrated only as high as 55°C. Here, we demonstrate a synthetic enzymatic pathway for acetone production that functions up to at least 70°C in vitro, made possible by the unusual thermostability of Adc from the mesophile C. acetobutylicum, and heteromultimeric acetoacetyl-CoA:acetate CoA-transferase (CtfAB) complexes from Thermosipho melanesiensis and Caldanaerobacter subterraneus, composed of a highly thermostable α-subunit and a thermally labile β-subunit. The three enzymes produce acetone in vitro at temperatures of at least 70°C, paving the way for bio-reactive distillation of acetone using a metabolically engineered extreme thermophile as a production host.


Pestalopyrones A-D, four tricyclic pyrone derivatives from the endophytic fungus Pestalotiopsis neglecta S3.
Four undescribed pyrone derivatives, pestalopyrones A-D, containing unusual tricyclic 5/6/6 polycyclic skeletons, were isolated from the ethyl acetate extract of the endophytic fungus Pestalotiopsis neglecta S3 derived from the fresh stems of Rubia podantha Diels (Rubiaceae). Their planar structures were elucidated mainly by NMR and HRESIMS. Pestalopyrones A-D contained six contiguous chiral carbons, and the relative configurations of C-4, C-5, and C-8 in tricyclic 5/6/6 polycyclic skeletons were determined by ROESY spectra. For pestalopyrone B, the absolute configuration of C-16 was determined by the Mosher's method. All isolated compounds were tested for their cytotoxicity against cancer cell lines, antibacterial activity, and inhibitory effect on the lipopolysaccharide (LPS)-induced nitric oxide (NO) production, and the results showed that pestalopyrone A inhibited LPS-induced NO production with an IC50 value of 35.8 μM.


Isolation of Enterococcus faecium, characterization of its antimicrobial metabolites and viability in probiotic Minas Frescal cheese.
The aim of this study was to isolate Enterococcus faecium from raw milk samples, to characterize its antimicrobial metabolites, and to evaluate its viability in a probiotic Minas Frescal cheese. For this, antagonist activity against Listeria monocytogenes, safety aspects and biochemical, genotypic, and probiotic characteristics of the isolates were evaluated. Minas Frescal cheese was manufactured with the isolate that showed the best characteristics in vitro, and its viability in the product was evaluated. It was observed that of the 478 lactic acid bacteria isolates, only isolate E297 presented antagonist activity, genes encoding for enterocin production and absence of virulence factors. Besides that, E297 presented probiotic characteristics in vitro, and maintained its viability (8.09 log CFU mL-1) for 14 days of cold storage, when it was added to cheese. Therefore, isolate E297 can be considered a promising microorganism for the manufacture of probiotic foods, especially Minas Frescal cheese.


Electrosorption and photocatalytic one-stage combined process using a new type of nanosized TiO₂/activated charcoal plate electrode.
In the present study, an activated charcoal (AC) plate was prepared by physical activation method. Its surface was coated with TiO₂ nanoparticles by electrophoretic deposition (EPD) method. The average crystallite size of TiO₂ nanoparticles was determined approximately 28 nm. The nature of prepared electrode was characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), and Brunauer-Emmett-Teller (BET) surface area measurement before and after immobilization. The electrosorption and photocatalytic one-stage combined process was investigated in degradation of Lanasol Red 5B (LR5B), and the effect of dye concentration, electrolyte concentration, pH, voltage, and contact time was optimized and modeled using response surface methodology (RSM) approach. The dye concentration of 30 mg L(-1), Na₂SO₄ concentration of 4.38 g L(-1), pH of 4, voltage of 250 mV, and contact time of 120 min were determined as optimum conditions. Decolorization efficiency increased in combined process to 85.65% at optimum conditions compared to 66.03% in TiO₂/AC photocatalytic, 20.09% in TiO₂/AC electrosorption, and 1.91% in AC photocatalytic processes.


Pyrimidine Triazole Thioether Derivatives as Toll-Like Receptor 5 (TLR5)/Flagellin Complex Inhibitors.
Protein-protein interactions have been regarded as "undruggable" despite their importance in many biological processes. The complex formed between host toll-like receptor 5 (TLR5) and flagellin, a globular protein that is the main component of a bacterial flagellum, plays a vital role in a number of pathogen defenses, immunological diseases and cancers. Through high-throughput screening, we identified two hits with a common pharmacophore, which were used to successfully develop a series of small-molecule probes as novel inhibitors of flagellin binding to TLR5. In a multitude of assays, 4-((4-benzyl-5-(pyridin4yl)-4H-1,2,4-triazol-3-yl)thio)pyrido[3',2':4,5]thieno[3,2-d]pyrimidine (TH1020) was identified as a potent antagonist of TLR5 signaling with promising activity (IC50 =0.85±0.12 μm) and specificity. Furthermore, TH1020 was shown to repress the expression of downstream TNF-α signaling pathways mediated by the TLR5/flagellin complex formation. Based on molecular docking simulation, TH1020 is suggested to compete with flagellin and disrupt its association with TLR5. TH1020 provides a much-needed molecular probe for studying this important protein-protein interaction and a lead compound for identifying novel therapeutics targeting TLR5.


Understanding Intimate Partner Violence in Jordan: Application of the Ecological Model.
To compare and contrast the Heise "ecological model of gender-based violence" with factors related to "intimate partner violence" (IPV) in Jordan. Findings of empirical studies examining IPV against Jordanian women were analyzed using the Heise ecological model (1998). Factors identified in the literature were generally consistent with the ecological model. However, the literature on IPV in Jordan rarely examined the role of community-level factors in the exosystem and sociopolitical factors in the macrosystem. Consequently, little is known about how these exosystem and macrosystem factors affect IPV in Jordan. The Heise model was then adapted to reflect identified factors in the Jordanian cultural context. More research aimed at uncovering barriers to identification and treatment of IPV targeting the exosystem and macrosystem is needed.


Multistep nucleation and multi-modification effect of Sc in hypoeutectic Al-Mg-Si alloys.
Structure of Al5Mg2SiMn casting alloy with and without of Sc addition, morphology of primary phases and specifically morphology of the Sc-containing intermetallic compounds and nanoscale properties of the α-Al matrix of Al-Mg-Si-Mn alloys were observed. Morphology and chemical composition were investigated by scanning electron microscopy on polished and deep etched microsections. It was found that addition of Sc changes morphology of α-Al, Al-Mg2Si eutectic and intermetallic compounds with Mn. α-Al dendrites change their morphology to spherical, lamellar Al-Mg2Si eutectic to fine fibrous eutectic. It was found that crystals Al3Sc can act as nucleating particles of α-Al grains, as well as Mg2Si eutectic fibrous. The mechanisms of nucleation can be attributed to heterogeneous nucleation.


Japanese encephalitis virus replicon-based vaccine expressing enterovirus-71 epitope confers dual protection from lethal challenges.
To construct safer recombinant flavivirus vaccine, we exploited Japanese encephalitis virus (JEV) replicon-based platform to generate single-round infectious particles (SRIPs) that expressed heterologous neutralizing epitope SP70 derived from enterovirus-71 (EV71). Such pseudo-infectious virus particles, named SRIP-SP70, although are not genuine viable viruses, closely mimic live virus infection to elicit immune responses within one round of viral life cycle. We found that, besides gaining of full protection to thwart JEV lethal challenge, female outbred ICR mice, when were immunized with SRIP-SP70 by prime-boost protocol, could not only induce SP70-specific and IgG2a predominant antibodies but also provide their newborns certain degree of protection against EV71 lethal challenge. Our results therefore exemplify that this vaccination strategy could indeed confer an immunized host a dual protective immunity against subsequent lethal challenge from JEV or EV71.


Outcomes of surgical treatments for acquired gynatresia in a tertiary institution in Ibadan, Nigeria.
A 16-year review of causes, clinical presentation and management outcomes of Acquired gynatresia (AG) at University College Hospital, Ibadan, Nigeria. Information was obtained using a proforma and data analysed using SPSS version 20.0. The mean age of the 31 women with AG was 35.6 ± 6.2 years. The majority (90.3%) were married and had a tertiary level of education. The mean parity was 0.74 ± 1.1 and 54.8% were nulliparous while 29.0% were primiparous women. The commonest cause of AG was the insertion of caustic substances into the vagina and this was mainly for the treatment of uterine fibroid (68.2%) and infertility (54.5%). Over three-quarters presented with dyspareunia while 54.8% had infertility. A majority (87.1%) had one-stage surgery with dissection/excision of fibrotic tissue being the commonest (45.2%) and sigmoid vaginoplasty the least performed. Postoperative complications were seen in 29.0% of cases. Acquired gynatresia remains a condition of public health interest despite an increase in female education and relatively improved health care in Nigeria.Impact statementWhat is already known on this subject? Acquired gynatresia (AG) could be of chemical and non-chemical origin and result from certain cultural beliefs and practices.What do the results of this study add? Irrespective of the women's level of education, there is inadequate awareness of the implications of inserting caustic materials into the vagina. However, there is a high success rate of treatment of AG.What the implications are of these findings for clinical practice and/or further research? There is a need to increase health education and awareness of the populace on the causes of AG and its associated complications.


Idiomarina rhizosphaerae sp. nov. isolated from rhizosphere soil of Kalidium cuspidatum, and reclassification of Idiomarina andamanensis as Pseudidiomarina andamanensis comb. nov., and Idiomarina mangrovi as Pseudidiomarina mangrovi comb. nov.
A Gram-stain-negative, strictly aerobic, gliding motile, non-spore forming, rod-shaped indole-3-acetic acid producing bacterial strain, designated M1R2S28T, was isolated from the rhizosphere soil of Kalidium cuspidatum, in Tumd Right Banner, Inner Mongolia, China. Strain M1R2S28T grew at pH 5.0-10.0 (optimum 8.0), 10-45 °C (optimum 37 °C), in the presence of 0-20% (w/v) NaCl (optimum 5%). The phylogenetic trees based on the 16S rRNA gene sequences and the core-genome both revealed that strain M1R2S28T clustered tightly with Idiomarina loihiensis L2-TRT, and shared 99.3, 99.2, 98.7, and < 98.7% of the 16S rRNA gene sequence similarities with I. loihiensis L2-TRT, I. abyssalis KMM 227 T, I. ramblicola R22T, and all the other current type strains. The strain contained ubiquinone-8 (Q-8) as the sole respiratory quinone. Its major polar lipids were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, an unidentified aminolipid, an unidentified phospholipid, and three unidentified lipids. Its major fatty acids were iso-C15:0, iso-C17:0, and iso-C17:1 ω9c. The genomic DNA G + C content was 46.8%. The average nucleotide identity based on BLAST (ANIb) and the digital DNA-DNA hybridization (dDDH) values of strain M1R2S28T to I. loihiensis L2-TRT, I. ramblicola R22T, and I. abyssalis KMM 227 T were 93.0, 82.9, and 81.8%, and 51.2, 26.0, and 25.0%, respectively. The phylogenetic and physiological results allowed the discrimination of strain M1R2S28T from its phylogenetic relatives. Idiomarina rhizosphaerae sp. nov. is, therefore, proposed with strain M1R2S28T (= CGMCC 1.19026 T = KCTC 92133 T) as the type strain. Additionally, based on the phylogenomic and genomic analysis results, Idiomarina andamanensis and Idiomarina mangrovi were transferred into genus Pseudidiomarina and be named Pseudidiomarina andamanensis comb. nov. and Pseudidiomarina mangrovi comb. nov., respectively.


What is new in preterm birth prevention? Important recent articles.
This month, we focus on current research in preterm birth prevention. Dr. Andrews discusses recent publications, and each is concluded with a "bottom line" that is the take-home message. The complete reference for each can be found in on this page, along with direct links to the abstracts.


Synthesis and properties of cyclic sandwich compounds.
Cyclic nanometre-scale sandwich complexes assembled from individual building blocks were synthesized. Sandwich complexes, in which a metal ion is π-coordinated by two planar aromatic organic rings belong to the foundations of organometallic chemistry. They have been successfully used in a wide variety of applications ranging from catalysis, synthesis and electrochemistry to nanotechnology, materials science and medicine1,2. Extending the sandwich structural motif leads to linear multidecker compounds, in which aromatic organic rings and metal atoms are arranged in an alternating fashion. However, the extension to a cyclic multidecker scaffold is unprecedented. Here we show the design, synthesis and characterization of an isomorphous series of circular sandwich compounds, for which the term 'cyclocenes' is suggested. These cyclocenes consist of 18 repeating units, forming almost ideally circular, closed rings in the solid state, that can be described by the general formula [cyclo-MII(μ-η8:η8-CotTIPS)]18 (M = Sr, Sm, Eu; CotTIPS = 1,4-(iPr3Si)2C8H62-). Quantum chemical calculations lead to the conclusion that a unique interplay between the ionic metal-to-ligand bonds, the bulkiness of the ligand system and the energy gain on ring closure, which is crucially influenced by dispersion interactions, facilitate the formation of these cyclic systems. Up to now, only linear one-dimensional multidecker sandwich compounds have been investigated for possible applications such as nanowires3-10. This textbook example of cyclic sandwich compounds is expected to open the door for further innovations towards new functional organometallic materials.


Changing Anesthesia Block Allocations Improves Endoscopy Suite Efficiency.
Non-operating room anesthesia (NORA) has grown and continues to expand as a proportion of all anesthesia practice in the United States [1, 2]. While many management processes have been adapted for NORA from the traditional operating room, it is still unclear what scheduling paradigm will maximize efficiency of resource utilization in this arena. In this study, we investigate the impact of tactical a shift from a shared group to individual, provider-specific block allocations for available anesthesia time in an endoscopy suite for adult patients undergoing elective endoscopy procedures at an academic hospital. Using a retrospective and prospective analysis, we measured elective time-in-block; elective time out-of-block; under-utilized (opportunity and non-opportunity unused) time; over-utilized time; and case tardiness to determine operational efficiency and clinical productivity. Over the study period, the monthly caseload remained constant. Elective time in block increased by 156% (p < 0.0001) and elective time out of block decreased by 38% (p < 0.0001). Opportunity unused time decreased by 28% (p < 0.0001) and productivity increased by 51% (p < 0.0001). Neither over-utilized time nor case tardiness showed a significant change after the intervention. Despite the evidence base supporting traditional approaches to anesthesia block allocation involving group block allocation and non-sequential case scheduling, we have demonstrated an advantage to individual block allocation in a GI endoscopy setting. This sequential case scheduling highlights how tactical decisions in NORA environments may require a rethinking of many practices that anesthesiologists have brought with them from the traditional OR. Using these efficiency and productivity metrics, further adjustments to scheduling practices should be investigated, and connecting these metrics to other systems outcomes, such as financial productivity, is an important next step as NORA services expand into the future.


Solid multifunctional granular bioink for constructing chondroid basing on stem cell spheroids and chondrocytes.
Stem cell spheroids are advanced building blocks to produce chondroid. However, the multi-step operations including spheroids preparation, collection and transfer, the following 3D printing and shaping limit their application in 3D printing. The present study fabricates an 'ALL-IN-ONE' bioink based on granular hydrogel to not only produce adipose derived stem cell (ASC) spheroids, but also realize the further combination of chondrocytes and the subsequent 3D printing. Microgels (6-10μm) grafted with β-cyclodextrin (β-CD) (MGβ-CD) were assembled and crosslinked byin-situpolymerized poly (N-isopropylacrylamide) (PNIPAm) to form bulk granular hydrogel. The host-guest action between β-CD of microgels and PNIPAm endows the hydrogel with stable, shear-thinning and self-healing properties. After creating caves, ASCs aggregate spontaneously to form numerous spheroids with diameter of 100-200μm inside the hydrogel. The thermosensitive porous granular hydrogel exhibits volume change under different temperature, realizing further adsorbing chondrocytes. Then, the granular hydrogel carrying ASC spheroids and chondrocytes is extruded by 3D printer at room temperature to form a tube, which can shrink at cell culture temperature to enhance the resolution. The subsequent ASC spheroids/chondrocytes co-culture forms cartilage-like tissue at 21 din vitro, which further matures subcutaneouslyin vivo, indicating the application potential of the fully synthetic granular hydrogel ink toward organoid culture.


Tracing Makerere University's 100 years' contribution to lung science/medicine.
Dear Editor, African Health Sciences Journal, This year, 2022, Makerere University will be celebrating 100 years of existence. As current lung science/medicine experts, we felt it as important to trace the University's contribution to lung science and medicine. In this letter, we trace and describe some of the early work done by Makerere University/Mulago Hospital affiliated scientists, identify prominent players in lung science over the 100 years, and present the university's scholarly contribution to this field, as available in online databases. We include both Makerere University and Mulago Hospital affiliated scientists, because for many years, staff of these two institutions have worked together in teaching, research and patient care.