Evaluation of the RadCalc collapsed cone dose calculation algorithm against measured data.
This work describes the experimental validation of the RadCalc (Lifeline software Inc, Tyler) collapsed cone dose calculation algorithm against measured data for a range of scenarios. 6 MV photon beam data were measured in a large water tank on a Varian TrueBeam linear accelerator. These were input into the RadCalc software, in conjunction with head geometry and output calibration information, then used to create a collapsed cone beam model. The model performance was assessed by comparison against measurement, using a selection of homogeneous and inhomogeneous geometries not incorporated into the original beam model. Dose calculations generated using the collapsed cone algorithm are generally in good agreement with measurement. However, the primary collimating of the linac is not accounted for in the RadCalc model and hence dose in the corners of large fields is significantly overestimated. Percentage depth doses were within 0.5% beyond a depth of 2 cm. The dose in the build-up region was underestimated by RadCalc Version 22.214.171.124, with (Distance To Agreement) discrepancies of up to 3 mm which were corrected in Version 126.96.36.199. Beam profiles for homogeneous phantom comparisons were within 2% in the central 80% of the field with out of field dose underestimated by no more than 3%. Dose comparisons in heterogeneous geometries were acceptable and generally within 3.5%. The largest observed differences were found at density interfaces and a result of the RadCalc dose resolution of 2 mm against 1 mm measured. Absolute dose comparisons demonstrated that RadCalc agreed with measurement to within 1.2% under homogeneous media irradiation geometries. For static beam IMRT deliveries agreement was within 2% or 2 mm of measured data, and for complex VMAT deliveries within 3% or 2 mm. The implementation of the (model-based) photon collapsed cone algorithm in RadCalc shows generally good agreement with measured data over a range of simple and complex scenarios considered.


Color stability and change in bioactive compounds of red beet juice concentrate stored at different temperatures.
In the present study, effect of storage temperature (25°, 35° and 45 °C) and light (with/without aluminum foil) on betalain content, color stability and bioactive compounds of concentrated red beet juice were investigated. Degradation of betalains and change in total phenolics content followed first-order kinetics while changes of L*, a*, b* and C* values and antioxidant capacity fitted zero-order kinetic. It was determined that the reaction rate constants of betacyanin and betaxanthin degradations in red beet juice concentrates increased with increasing storage temperature and time. The activation energies for betaxanthin degradation (Ea: 92.04-93.27 kJ mol-1) in comparison with the activation energies for betacyanin degradation (Ea: 66.07-66.13 kJ mol-1) in all samples demonstrate that susceptibility to temperature of betaxanthin is higher than that of betacyanin. According to L* and a* parameters it can be suggested that color stability in red beet juice concentrate stored with aluminum foil found better than that in the sample without aluminum foil. However, there was no significant difference (p > 0.05) between samples with and without foil as regards changes in total phenolics, betalains and antioxidant capacity. In addition, 25 °C can be proposed for providing betalain stability of red beet juice concentrates during storage.


Prediction errors drive dynamic changes in neural patterns that guide behavior.
Learning describes the process by which our internal expectation models of the world are updated by surprising outcomes (prediction errors [PEs]) to improve predictions of future events. However, the mechanisms through which error signals dynamically influence existing neural representations are unknown. Here, we use functional magnetic resonance imaging (fMRI) in humans solving a two-step Markov decision task to investigate changes in neural activation patterns following PEs. Using a dynamic multivariate pattern analysis, we can show that PE-related fMRI responses in error-coding regions predict trial-by-trial changes in multivariate neural patterns in the orbitofrontal cortex, the precuneus, and the ventromedial prefrontal cortex (vmPFC). Importantly, the dynamics of these pattern changes in the vmPFC also predicted upcoming changes in choice strategies and thus highlight the importance of these pattern changes for behavior.


Affinity chromatography of thiol-containing purines and ribonucleic acid.
Evidence is presented for the specific interaction of 6-mercaptopurine with mercurated cellulose. Following from this, a new method is described for the affinity chromatography of thiol-containing molecules and of RNA containing incorporated 6-thioguanosine on columns of mercurated cellulose. This technique may find application in the study of RNA metabolism and gene expression.


Differences in mortality rate between abrupt and progressive carotid ligation in the gerbil: role of endogenous angiotensin II.
Studies have shown that in comparison to rapid occlusion of a vessel, gradual occlusion produces less severe tissue ischemia due to a more effective development of collateral circulation. As other studies have shown that collateral circulation can be enhanced by stimulation of the endogenous renin-angiotensin II system, it was hypothesized that this system is involved in the mechanism of protection against ischemia that obtains during gradual vascular occlusion. To test this hypothesis, mortality rates were evaluated in gerbils subjected to gradual vascular occlusion by means of progressive carotid ligation while simultaneously infused with inhibitors of the renin-angiotensin II cascade--enalaprilat or saralasin. Groups of animals with either abrupt or progressive carotid ligation infused with saline served as controls. Results showed that (1) in saline-infused animals, there was a significant decrease in the mortality rate of progressive-ligated animals when compared to abrupt-ligated animals, and (2) administration of either enalaprilat or saralasin to progressive-ligated animals resulted in mortality rates that were indistinguishable from those of saline-infused abrupt-ligated animals. These results suggest that the endogenous renin-angiotensin system is indeed involved in an adaptive mechanism that occurs during progressive ligation of the carotid artery, and more specifically, that the relatively benign effect of progressive carotid ligation may be due to the action of angiotensin II to stimulate the development of collateral circulation and reduce the severity of focal brain ischemia.


Identification of an IGF-1R kinase regulatory phosphatase using the fission yeast Schizosaccharomyces pombe and a GFP tagged IGF-1R in mammalian cells.
To study the regulation of type 1 insulin like growth factor receptor (IGF-1R) tyrosine kinase activity using the fission yeast Schizosaccharomyces pombe and a green fluorescent protein (GFP) tagged, full length IGF-1R. The beta chain of the IGF-1R (betawt) was expressed under inducible conditions in the fission yeast S. pombe. Western blot analysis with antiphosphotyrosine antibodies was used to assess the kinase activity of betawt. A GFP tagged IGF-1R (GFP-IGF-1R) was constructed to study the tyrosine kinase activity of the full length IGF-1R. The signalling capabilities of GFP-IGF-1R in response to IGF-1 stimulation were investigated in transiently transfected fibroblasts. Immunofluorescent staining for cellular phosphotyrosine content was used to assess the localisation and tyrosine kinase activity of GFP-IGF-1R. The betawt protein displayed functional tyrosine kinase activity in S pombe and phosphorylated endogenous yeast proteins. In response to IGF-1 stimulation, the GFP-IGF-1R became autophosphorylated and also activated the phosphatidylinositol 3-kinase and mitogen activated protein kinase pathways. Tyrosine phosphorylation and kinase activity of the GFP-IGF-1R could be visualised by immunofluorescence with antiphosphotyrosine antibodies. Coexpression of a mammalian tyrosine phosphatase PTP1B with betawt completely inhibited this tyrosine kinase activity in yeast and also reduced the tyrosine phosphorylation in COS cells transfected with the GFP-IGF-1R. Schizosaccharomyces pombe can be used to analyse the tyrosine kinase activity of the IGF-1R beta chain and its regulation by tyrosine phosphatases. In addition, the regulation of IGF-1R tyrosine kinase activity can be studied using a GFP tagged IGF-1R. Using both of these methods, IGF-1R kinase activity was shown to be inhibited by the protein tyrosine phosphatase, PTP1B.


Demographic factors associated with smoking cessation during pregnancy in New South Wales, Australia, 2000-2011.
Smoking during pregnancy increases the risk of adverse health outcomes for both the mother and the child. Rates of smoking during pregnancy, and rates of smoking cessation during pregnancy, vary between demographic groups. This study describes demographic factors associated with smoking cessation during pregnancy in New South Wales, Australia, and describes trends in smoking cessation in demographic subgroups over the period 2000 - 2011. Data were obtained from the New South Wales Perinatal Data Collection, a population-based surveillance system covering all births in New South Wales. Multivariate logistic regression was used to explore associations between smoking cessation during pregnancy and demographic factors. Between 2000 and 2011, rates of smoking cessation in pregnancy increased from 4.0% to 25.2%. Demographic characteristics associated with lower rates of smoking cessation during pregnancy included being a teenage mother, being an Aboriginal person, and having a higher number of previous pregnancies. Between 2000 and 2011, rates of smoking cessation during pregnancy increased dramatically across all demographic groups. However, specific demographic groups remain significantly less likely to quit smoking, suggesting a need for targeted efforts to promote smoking cessation in these groups.


Hypnosis with specific relation to biofeedback and behavior therapy. Theoretical and clinical considerations.
Hypnosis as an intrapsychological and interpersonal experience is used as an integrative and amplifying procedure in relation to biofeedback mechanism and behavior therapy. The hypnotic capacity for linking cognitive to affective reactions within a feedback loop of sensory and motor imagery is presented as a dynamic approach to behavior modification during psychotherapy.


Magnetic resonance imaging study of the sciatic nerve variation in the pediatric gluteal region: Implications for the posterior approach of the sciatic nerve blockade.
In pediatric patients, the sciatic nerve is one of the most commonly blocked peripheral nerves during orthopedic procedures of the lower limb. Ultrasound guidance is the current standard for a successful localization of the sciatic nerve in the gluteal region. Relevant anatomical landmarks are also used to determine the nerve location when ultrasound is not available or inadequate. However, reports have demonstrated paucity of information regarding the sciatic nerve location and variation in the hip throughout pediatric development. This imaging study aimed to document and analyze the relative morphometric relationship of the sciatic nerve in the pediatric gluteal region throughout development. The location of the sciatic nerve in relation to bony landmarks was measured in 84 pediatric magnetic resonance imaging of patients aged 0.7-15.8 years. The sciatic nerve was identified medial to the most lateral point of greater trochanter at the level of ischial spine and the tip of coccyx. The strong positive correlation between sciatic nerve to landmark distances and age and stature demonstrated linear variation between sciatic nerve location with age and growth of children. To predict the nerve location in the gluteal region, regression equations using patient age were created, having implications for the posterior approach of the sciatic nerve blockade in children. Clinically significant differences were found between sexes, specifically in the older age group. Despite the small sample size of younger age group, this study is the first to document the morphometric changes of the sciatic nerve in the gluteal region across pediatric development and may be useful for providing confirmatory guidelines for nerve location when ultrasound is not accessible or cannot be utilized for practice.


Water vapor thermal therapy to alleviate catheter-dependent urinary retention secondary to benign prostatic hyperplasia.
Water vapor thermal therapy utilizes convectively delivered thermal energy to target ablation of obstructive prostatic tissue. We report results of this thermal therapy for relief of nonneurogenic complete urinary retention associated with BPH. We conducted a retrospective analysis of 38 catheter-dependent men with complete urinary retention consecutively enrolled in a registry in two centers: median age 75.5 years and multiple comorbidities, median prostate volume 58.5 cc (23-153), median 2 failed trials without catheter (TWOCs), and median catheter dependency 3 months (0.3-35). The Rezūm™ System thermal therapy procedure was performed in an ambulatory surgery center with conscious sedation or an office procedure room with a modified periprostatic block. Water vapor injections were customized to the configuration of the hyperplastic gland, including median lobe and/or enlarged central zone. Of the 38 treated patients, one was lost to follow-up and 26 of 37 (70.3%) voided spontaneously (mean of 1.6 ± 0.8 TWOCs) and were catheter free a median of 26 days (range 4-65) after the procedure; 18 of these 26 (69%) patients discontinued BPH medications. No significant differences in age, prostate volume, number of water vapor injections, or presence of the median lobe were associated with predicting a successful treatment outcome. Duration of follow-up for 20 catheter-free patients was a median of 475 days or 15.8 months (140-804 days); six patients were followed a median of 31.5 days (0-60). Adverse events were infrequent, mild, and resolved quickly including dysuria in five patients (13%), gross hematuria in four (10.5%), and UTIs in two (2.6%) with indwelling catheters. Water vapor thermal therapy may provide an effective and safe alternative to surgical treatment in this group of catheter-dependent patients in complete urinary retention.


Associations of blood biomarkers with arterial stiffness in patients with diabetes mellitus: A population-based study.
Arterial stiffness contributes to additional cardiovascular risks in diabetic patients by triggering the loss of vascular and myocardial compliance and promoting endothelial dysfunction. Thus, prevention of arterial stiffness is a public health priority, and the identification of potential biomarkers may provide benefits for early prevention. This study investigates the relationships between serum laboratory tests and pulse wave velocity (PWV) tests. We also investigated the associations between PWV and all-cause mortality. We examined a panel of 33 blood biomarkers among diabetic populations in the Atherosclerosis Risk in Communities Study. The carotid-femoral (cfPWV) and femoral-ankle PWV (faPWV) were measured using an automated cardiovascular screening device. The aortic-femoral arterial stiffness gradient (afSG) was calculated as faPWV divided by cfPWV. Biomarker levels were log-transformed and correlated with PWV. Cox proportional hazard models were employed for survival analysis. Among 1079 diabetic patients, biomarkers including high-density lipoprotein cholesterol, glycated hemoglobin, high-sensitivity troponin T, cystatin C, creatinine, and albuminuria were significantly correlated with afSG (R = 0.078, -0.193, -0.155, -0.153, -0.116, and -0.137, respectively) and cfPWV (R = -0.068, 0.175, 0.128, 0.066, 0.202, and 0.062, respectively). Compared with the lowest tertile of afSG, the risk of all-cause mortality was lower in the highest tertile (hazard ratio 0.543; 95% confidence interval 0.328-0.900). Certain biomarkers related to blood glucose monitoring, myocardial injury, and renal function significantly correlated with PWV, suggesting that these putative risk factors are likely to be important atherosclerosis mechanisms in diabetic patients. AfSG may be an independent predictor of mortality among diabetic populations.


A Raman spectroscopic study of the uranyl sulphate mineral johannite.
Raman spectroscopy at 298 and 77K has been used to study the secondary uranyl mineral johannite of formula (Cu(UO2)2(SO4)2(OH)2 x 8H2O). Four Raman bands are observed at 3593, 3523, 3387 and 3234cm(-1) and four infrared bands at 3589, 3518, 3389 and 3205cm(-1). The first two bands are assigned to OH- units (hydroxyls) and the second two bands to water units. Estimations of the hydrogen bond distances for these four bands are 3.35, 2.92, 2.79 and 2.70 A. A sharp intense band at 1042 cm(-1) is attributed to the (SO4)2- symmetric stretching vibration and the three Raman bands at 1147, 1100 and 1090cm(-1) to the (SO4)2- anti-symmetric stretching vibrations. The nu2 bending modes were at 469, 425 and 388 cm(-1) at 77K confirming the reduction in symmetry of the (SO4)2- units. At 77K two bands at 811 and 786 cm(-1) are attributed to the nu1 symmetric stretching modes of the (UO2)2+ units suggesting the non-equivalence of the UO bonds in the (UO2)2+ units. The band at 786cm(-1), however, may be related to water molecules libration modes. In the 77K Raman spectrum, bands are observed at 306, 282, 231 and 210cm(-1) with other low intensity bands found at 191, 170 and 149cm(-1). The two bands at 282 and 210 cm(-1) are attributed to the doubly degenerate nu2 bending vibration of the (UO2)2+ units. Raman spectroscopy can contribute significant knowledge in the study of uranyl minerals because of better band separation with significantly narrower bands, avoiding the complex spectral profiles as observed with infrared spectroscopy.


Reversion of mouse postimplantation epiblast stem cells to a naïve pluripotent state by modulation of signalling pathways.
Mouse postimplantation epiblast cultured in activin and basic fibroblast growth factor gives rise to continuously growing epiblast stem cells (EpiSCs) that share key properties with postimplantation epiblast, such as DNA methylation and an inactive X-chromosome. EpiSCs also show a distinct gene expression profile compared to embryonic stem cells (ESCs) derived from preimplantation blastocysts, and do not contribute efficiently to chimeras. EpiSCs can, however, revert to pluripotent ESC-like cells upon exposure to leukemia inhibitory factor-Stat3 signalling on feeder cells. Here we describe a protocol for the establishment of EpiSCs and their reversion to ESCs.


Population-based HPV vaccination programmes are safe and effective: 2017 update and the impetus for achieving better global coverage.
Persistent oncogenic human papillomavirus (HPV) is the cause of cervical cancer, as well as cancers of the anus, penis, vulva, vagina and oropharynx. There is good evidence that prophylactic HPV vaccines are immunogenic and effective against targeted-type HPV infections and type-specific genital lesions, including high-grade cervical intraepithelial neoplasia (CIN), when administered prior to HPV infection. There is good evidence that HPV vaccines are safe in population usage, with the most frequent adverse event being injection-site reactions. There is evidence to support some cross-protection against non-targeted types occurring following the administration of HPV vaccines. There is limited evidence suggesting that HPV vaccines may be beneficial in preventing future disease in women treated for high-grade CIN. This chapter focuses on the accumulated evidence regarding the global use of the three licensed HPV vaccines including safety, immunogenicity, duration of protection, effectiveness, coverage to date and barriers to higher coverage.


Latent Tuberculosis Therapy Outcomes in Dialysis Patients: A Retrospective Cohort.
Maintenance dialysis patients are at an increased risk for active tuberculosis (TB). In 2012, British Columbia, Canada, began systematically screening maintenance dialysis patients for latent TB infection (LTBI) and treating people with evidence of LTBI when appropriate. We examined LTBI treatment outcomes and compared treatment outcomes before and after rollout of the systematic screening program. Retrospective cohort study. The study comprised 365 people in British Columbia, Canada, initiating at least 90 days of dialysis from January 1, 2001, to May 31, 2017, and starting LTBI therapy: 290 (79.5%) people in the recent cohort and 75 (20.5%) in the historical cohort. People starting LTBI therapy from January 1, 2012, onward were classified as the recent cohort, whereas people starting LTBI therapy before January 1, 2012, were classified as the historical cohort. Systematic LTBI screening and therapy. Proportion of people who experience grade 3 to 5 adverse events (AEs) or any grade rash and end-of-treatment outcomes. Outcomes were reported using descriptive statistics. 2-sample test of proportions using χ2 distribution was used to test for statistical significance between the recent and historical cohorts. 298 (81.6%) people successfully completed LTBI therapy. The proportion of people experiencing a grade 3 to 4 AE or any grade rash was 21.1%. Most AEs were related to gastrointestinal events, general malaise, or pruritus that resulted in regimen changes. 2 (0.5%) people were hospitalized for AEs related to LTBI therapy. No significant difference was found between the recent and historical cohorts in all outcomes of interest. No grade 5 AEs (deaths) were attributed to LTBI therapy. Retrospective data and generalizability outside low-TB-burden settings. Our findings suggest that a high proportion of people receiving maintenance dialysis can complete LTBI therapy. The rate of grade 3 to 4 AEs was high and associated with frequent medication changes during therapy. LTBI therapy in maintenance dialysis may be safe but requires close monitoring.


School-based interventions to enhance knowledge and improve case management of schistosomiasis: a case study from Hunan, China.
This paper discusses an intersectoral health-related intervention, using cartoons and video-recording, print materials and face-to-face educational methods, to increase children's knowledge of schistosomiasis, which in turn might improve the case management of early diagnosis and treatment. The main components of the project were (i) the collaboration between the departments of public health and education and (ii) a randomized, controlled, school-based field trial conducted in the Dongting Lake region, China. Children in the experimental group (n=604) and their parents participated in the educational programme. Control children (n=527) received a 2 hour lecture about the disease. All participants were pre-tested, and retested five months after the conduct of the educational intervention. The results show significant changes among children and their parents in the experimental group related to knowledge about schistosomiasis and beliefs towards screening and treatment of the disease. Children in the experimental group also had better compliance than children in the control group for regular screening for schistosomiasis. These findings indicate that carefully designed education programmes are useful for providing both children and their families with information about the prevention and treatment of schistosomiasis. Intersectoral collaboration holds promise to deliver research-based interventions for enhanced knowledge of schistosomiasis and improved case management.


The mouse Formin mDia1 is a potent actin nucleation factor regulated by autoinhibition.
Formin proteins are widely expressed in eukaryotes and play essential roles in assembling specific cellular actin-based structures. Formins are defined by a Formin Homology 2 (FH2) domain, as well as a proline-rich FH1 domain that binds the actin monomer binding protein, profilin, and other ligands. Constructs including FH2 of budding yeast Bni1 or fission yeast Cdc12 formins nucleate actin filaments in vitro. In this study, we demonstrate that FH2-containing constructs of murine mDia1 (also called p140 mDia or Drf1) are much more potent actin nucleators than the yeast formins. FH1 is necessary for nucleation when actin monomers are profilin bound. mDia1 is a member of the Diaphanous formin subfamily (Dia), whose members contain an N-terminal Rho GTPase binding domain (GBD) and a C-terminal Diaphanous autoinhibitory domain (DAD, ). Based on cellular and in vitro binding studies, an autoinhibitory model for Dia formin regulation proposes that GBD binding to DAD inhibits Dia-induced actin remodeling, whereas Rho binding activates by releasing GBD from DAD. Supporting this model, our results show that an N-terminal mDia1 construct strongly inhibits actin nucleation by the C terminus. RhoA partially relieves inhibition but does so when bound to either GDP or GTP analogs. Both N- and C-terminal mDia1 constructs appear to be multimeric.


Burden of Proteinuria and Risk Factors of Chronic Kidney Disease among Adult Population in Urban Puducherry, India.
In the recent times, Chronic Kidney Diseases (CKDs) are emerging as a serious problem all over the world along with diabetes mellitus and hypertension. The presence of proteinuria is considered as an indicator of increased risk of progressive kidney diseases. To determine the prevalence of proteinuria among an adult population of a tertiary care institute of Puducherry, India. A cross-sectional study was carried out in the field practice areas of an urban health centre of a tertiary care institute, in Puducherry, India. A total of 215 study respondents were selected by systematic random sampling. All adults aged above 18 years who were residing for at least a year in Puducherry were included in the study. The study period was from July 2015 to October 2015. All the categorical variables were described as proportions. Chi square test was done to compare between two proportions. Univariate analysis was done to estimate the Odds Ratio (OR) with 95% CI. The mean age of the study participants was 38.5±12.8 years. Majority, 145 (67.4%) of the study participants were females. The prevalence of proteinuria was found to be 9.3%. While 4.7% and 11.2% of participants used tobacco and alcohol respectively, 13.5% and 27.9% had diabetes mellitus and hypertension respectively. Elderly age, diabetes mellitus and hypertension were found to be statistically significant predictors for proteinuria. The prevalence of proteinuria was high in our study population (9.3%) and hypertension and diabetes mellitus were also found to be risk factors for CKD. Routine screening among the general population for proteinuria in community-based settings might be an effective step to bring down the rate of progression of CKD.


The use of NMR chemical shifts to analyse the MD trajectories: simulation of bovine pancreatic trypsin inhibitor dynamics in water as a test case for solvent influences.
In this paper the NMR secondary chemical shifts, that are estimated from a set of 3D-structures, are compared with the observed ones to appraise the behaviour of a known x-ray diffraction structure (of the bovine pancreatic trypsin inhibitor protein) when various molecular dynamics are applied. The results of a 200 ps molecular dynamics under various conditions are analysed and different ways to modify the molecular dynamics are considered. With the purpose of avoiding the time-consuming explicit representation of the solvent (water) molecules, an attempt was made to understand the role of the solvent and to develop an implicit representation, which may be refined. A simulation of hydrophobic effects in an aqueous environment is also proposed which seems to provide a better approximation of the observed solution structure of the protein.


[Current status of surgical treatment of peritoneal metastases from colorectal cancer].
Cytoreductive surgery, often in combination with hyperthermic intraperitoneal chemotherapy (HIPEC), has been instrumental in improving the survival of patients with peritoneal metastases from colorectal cancer. Recent studies have highlighted the benefits of complete cytoreduction, while the role of the HIPEC treatment remains unclear. An oxaliplatin-based HIPEC over 30 min could not achieve any clear benefits in studies on colorectal cancer, neither in the therapeutic nor in the prophylactic setting, but caused relevant side effects and increased the morbidity. The negative results of these studies with respect to oxaliplatin-based HIPEC require critical appraisal; however, they should by no means be regarded as a general setback for surgical treatment of peritoneal metastases and be misunderstood as a general failure of this treatment. While HIPEC after complete surgical cytoreduction of peritoneal metastases from colorectal cancer requires further research, cytoreductive surgery should still be regarded as a highly effective treatment for suitable patients with limited abdominal tumor dissemination.


The correlation between immune subtypes and consensus molecular subtypes in colorectal cancer identifies novel tumour microenvironment profiles, with prognostic and therapeutic implications.
Solid tumour growth is the consequence of a complex interplay between cancer cells and their microenvironment. Recently, a new global transcriptomic immune classification of solid tumours has identified six immune subtypes (ISs) (C1-C6). Our aim was to specifically characterise ISs in colorectal cancer (CRC) and assess their interplay with the consensus molecular subtypes (CMSs). Clinical and molecular information, including CMSs and ISs, were obtained from The Cancer Genome Atlas (TCGA) (N = 625). Immune cell populations, differential gene expression and gene set enrichment analysis were performed to characterise ISs in the global CRC population by using CMSs. Only 5 ISs were identified in CRC, predominantly C1 wound healing (77%) and C2 IFN-γ dominant (17%). CMS1 showed the highest proportion of C2 (53%), whereas C1 was particularly dominant in CMS2 (91%). CMS3 had the highest representation of C3 inflammatory (7%) and C4 lymphocyte depleted ISs (4%), whereas all C6 TGF-β dominant cases belonged to CMS4 (2.3%). Prognostic relevance of ISs in CRC substantially differed from that reported for the global TCGA, and ISs had a greater ability to stratify the prognosis of CRC patients than CMS classification. C2 had higher densities of CD8, CD4 activated, follicular helper T cells, regulatory T cells and neutrophils and the highest M1/M2 polarisation. C2 had a heightened activation of pathways related to the immune system, apoptosis and DNA repair, mTOR signalling and oxidative phosphorylation, whereas C1 was more dependent of metabolic pathways. The correlation of IS and CMS allows a more precise categorisation of patients with relevant clinical and biological implications, which may be valuable tools to improve tailored therapeutic interventions in CRC patients.


Toenails as an alternative source material for the extraction of DNA from decomposed human remains.
The DNA identification of decomposed human remains for coronial investigations at the Victorian Institute of Forensic Medicine routinely requires the retrieval and processing of a bone sample obtained from the deceased. Bone is a difficult sample type to work with as it requires surgical removal from the deceased, refrigerated storage, and additional processing steps prior to DNA analysis in comparison to other samples types such as buccal swabs or blood stains. In an attempt to overcome the issues posed by bone, a DNA extraction method utilising toenails as an alternate source material was optimised and trialled. Two DNA extraction methods were optimised for digestion of toenail material, with the method utilising the QIAGEN DNA Investigator Kit selected for a casework trial. Single source DNA profiles, matching those of the conventional samples taken, were obtained for toenail samples collected from 28 of 30 coronial cases available for this study. Of these, 26 toenail samples produced full profiles. Although the overall DNA profile quality from the toenails was less than that of the conventional sample, the profiles from toenails met the reporting requirements for identification. Based on the results obtained, the Victorian Institute of Forensic Medicine will be implementing toenails as the primary sample type for collection from decomposed remains when blood is not a suitable sample type.


The newly isolated lytic bacteriophages st104a and st104b are highly virulent against Salmonella enterica.
To screen Irish faecal samples from a variety of sources with a view to isolating novel anti-Salmonella phages and to subsequently evaluate their lytic capability. Two novel anti-Salmonella phages st104a and st104b were isolated from a screening programme based on their lytic capability. The phages produced significantly larger plaques (2 mm) on the chosen indicator Salmonella enterica strain, DPC6046, when compared with the well-known control phage, Felix 01 (0.5 mm). Both phages st104a and st104b were found to have a broad host range within the Salm. enterica species. During in vitro trials, both phages (st104a and st104b) reduced Salm. enterica numbers more than 99% within 1 h. In vivo studies, involving the addition of the phage to porcine gastric juice (pH 2.5) demonstrated that phage st104a and phage Felix 01 were capable of surviving (10 and 30% survival respectively) the acidic conditions, unlike st104b, which was undetectable after 2 h exposure. Two novel lytic anti-Salmonella phages were isolated and characterized. With the exception of phage Felix 01, there has been relatively little phage therapy work performed using lytic Salmonella phage. In this study, the lytic phages st104a and st104b were isolated as a result of a faecal screening programme. Subsequently, phage st104a was found to have potential for biocontrol of Salm. enterica numbers if administered orally to pigs given their survival in porcine gastric juice, whereas, phage st104b may have potential in reducing cell numbers if applied by alternative approaches.


Dealloying fabrication of hierarchical porous Nickel-Iron foams for efficient oxygen evolution reaction.
Designing and preparing highly active oxygen evolution reaction (OER) electrodes are essential for improving the overall efficiency of water splitting. Increasing the number of active sites is the simplest way to enhance OER performance. Herein, we present a dealloy-etched Ni-Fe foam with a hierarchical nanoporous structure as integrated electrodes with excellent performance for OER. Using the dealloying method on the Ni-Fe foam framework, a nanoporous structure is produced, which is named nanoporous Ni-Fe@Ni-Fe foam (NP-NF@NFF). Because of the peculiarities of the dealloying method, the NP-NF@NFF produced contains oxygen vacancies and heterojunctions. As a result, NP-NF@NFF electrode outperforms state-of-the-art noble metal catalysts with an extremely low overpotential of 210 and 285 mV at current densities of 10 and 100 mA cm-2, respectively. Additionally, the NP-NF@NFF electrode shows a 60-h stability period. Therefore, NP-NF@NFF provides new insights into the investigation of high-performance transition metal foam electrodes with effective active sites for efficient oxygen evolution at high current densities.


Association of Retinal Blood Flow with Progression of Visual Field in Glaucoma.
In the glaucoma clinic, patients with normal intraocular pressure (IOP) can sometimes show visual field (VF) progression. Therefore, clarification of relationship between vascular status and glaucomatous VF deterioration is a focus of interest. We used optical coherence tomography angiography (OCTA), with the aim of evaluating the relationship between vessel density (VD) and VF progression in glaucoma patients. We included 104 eyes with open angle glaucoma who were followed up for at least 5 years in this retrospective case-control study. Superficial and deep VD of macula were assessed by OCTA. Regression analysis and Cox proportional hazards model were used to identify factors significantly associated with VF progression. In logistic regression analysis determining VF progression from Guided Progression Analysis (GPA) program, initial IOP and deep macular VD were significantly associated with VF progression in multivariate analysis (P = 0.019 and 0.004). Cox proportional hazards model also identified deep macular VD as significantly related to VF progression (P = 0.035). In conclusion, initial IOP and deep VD were related to VF deterioration in glaucoma. Deep VD might be used as a surrogate of glaucomatous VF progression related with vascular incompetence.


Meiosis study in a population sample from Afghanistan: allele frequencies and mutation rates of 16 STR loci.
The 16 short tandem repeat systems D3S1358, VWA, FGA, TH01, TPOX, CSF1PO, D5S818, D13S317, D7S820, ACTBP2, D2S1338, D16S539, D19S433, D21S11, D18S51 and D8S1179 were amplified in a population sample composed of 333 immigrants from Afghanistan. The 16 loci met Hardy-Weinberg expectations and possess a combined matching probability of 1 in 3.6 x 10(14) and a combined mean exclusion chance greater than 0.9996 in this Afghan population. Approximately 12,000 meiotic transfers were investigated and 19 mutations were observed in the repeat units of FGA (n=6), ACTBP2 (n=5), D3S1358 (n=2), D5S818 (n=2), D7S820 (n=2), VWA (n=1) and D8S1179 (n=1).


Craniospinal irradiation for acute lymphoblastic leukemia with central nervous system disease at diagnosis: a report from the Children's Cancer Group.
This study attempted to determine if central nervous system (CNS) disease at diagnosis is a poor prognostic factor in children with acute lymphoblastic leukemia (ALL) and whether 6 Gy of spinal irradiation is an adequate dose for these patients. Previously the Children's Cancer Group (CCG) treated patients with ALL and CNS disease at diagnosis with cranio (24 Gy)-spinal (12 Gy) irradiation, as well as systemic and intrathecal chemotherapy. In a series of CCG trials completed in 1989 the spinal dose was empirically reduced to 6 Gy for patients receiving systemic chemotherapy with an intensive consolidation phase to limit hematopoietic toxicity. The spinal dose was left at 12 Gy for patients treated with a less intensive consolidation phase. With a median follow-up for surviving patients of 74 months, the 5-year event-free survival for 53 patients with CNS disease at diagnosis was 69 +/- 13% (+/- 2 standard deviations), similar to the value obtained for 3364 patients without CNS disease, 67 +/- 2%. Corresponding values for 5-year survival were 77 +/- 12% and 80 +/- 1%, and for freedom from isolated first CNS relapse, were 90 +/- 9% and 94 +/- 1%. Event-free survival, survival, and freedom from isolated first CNS relapse in the 6-Gy group were as good as in the 12-Gy group. CNS disease at diagnosis is not a poor prognostic factor for children with ALL who are treated with intensive systemic chemotherapy, craniospinal irradiation, and intrathecal chemotherapy. Six Gy is an adequate dose of spinal irradiation for these patients.


Coordinating synaptic growth without being a nervous wreck.
The function and regulation of actin-cytoskeletal dynamics during synaptic growth is poorly understood. In this issue of Neuron, Coyle et al. report the identification of nervous wreck (nwk), a synapse-specific adaptor molecule in Drosophila that regulates synaptic growth and morphology via Wasp, a well-characterized mediator of actin dynamics.


Real-time reliable determination of binding kinetics of DNA hybridization using a multi-channel graphene biosensor.
Reliable determination of binding kinetics and affinity of DNA hybridization and single-base mismatches plays an essential role in systems biology, personalized and precision medicine. The standard tools are optical-based sensors that are difficult to operate in low cost and to miniaturize for high-throughput measurement. Biosensors based on nanowire field-effect transistors have been developed, but reliable and cost-effective fabrication remains a challenge. Here, we demonstrate that a graphene single-crystal domain patterned into multiple channels can measure time- and concentration-dependent DNA hybridization kinetics and affinity reliably and sensitively, with a detection limit of 10 pM for DNA. It can distinguish single-base mutations quantitatively in real time. An analytical model is developed to estimate probe density, efficiency of hybridization and the maximum sensor response. The results suggest a promising future for cost-effective, high-throughput screening of drug candidates, genetic variations and disease biomarkers by using an integrated, miniaturized, all-electrical multiplexed, graphene-based DNA array.


Gonadal thermoluminiscence dosimetry in cranial computed tomography with the EMI scanner.
A cranial computed tomographic examination (CT) consisting of four scans with the water bag equipped Mark I EMI scanner will expose the adult male gonads to a radiation dose of approximately 2 milliröntgen (mR).


Invertebrate health in marine protected areas (MPAs).
Marine protected areas (MPAs) consist of various categories of safeguarded areas in the marine environment, from semi-protected areas to total no take zones. The reported effects of MPAs are overwhelmingly positive, with numerous reports of fish size (biomass), abundance (recovery) and diversity increases, however, literature is lacking on the role and consequences of MPAs on parasite and disease dynamics, and in particular, invertebrate health. The implementation of MPAs has been known to alter trophic cascades and community dynamics, and with invertebrates commonly at the base of these systems, it is vital that their status is investigated. Overcrowding in areas closed to fishing is known to have parasitological consequences in some scenarios, and land/water use change has been known to alter host and vector communities, possibly elevating disease risk. Equally, reserves can be used as tools for alleviating impacts of marine disease. This review aims to consolidate extant literature and provide a comprehensive viewpoint on how invertebrates (and their health status) can be affected by MPAs, which are increasingly being implemented based on the relative urgency now being placed on protecting global biodiversity. In highlighting the paucity of knowledge surrounding MPAs and disease, especially that of the unenigmatic invertebrate groups, this review, published in the Special Issue on 'Invertebrates as One Health Sentinels', provides an opportunity for wide dissemination and provocation of further research in this area.


Substance P and serotonin immunoreactivity in the rat inferior olive.
The present study analyzes in detail the distribution of substance P and serotonin immunoreactivity in the inferior olivary complex of the rat using the indirect antibody peroxidase-antiperoxidase technique. The entire dorsal accessory olive contains an abundance of substance P fibers and varicosities. In addition, the dorsal cap of Kooy, nucleus Beta, parts b and c of the medial accessory olive, and the dorsomedial cell column contain numerous substance P immunoreactive elements. Fibers and varicosities positive for serotonin are present throughout the entire dorsal accessory olive. In addition, the rostral medial accessory olive contains an abundance of serotonin immunostaining. Scattered serotonin immunoreactive elements are present in the principal olive in particular in the area immediately subjacent to the lateral aspect of the dorsal accessory olive. Only scant serotonin immunolabeling is present in the dorsal cap of Kooy, nucleus Beta and the dorsomedial cell column. Speculations on possible functional roles of these two putative neurotransmitters are presented.


Effect of different metabolic pathways on itaconic acid production in engineered Corynebacterium glutamicum.
Itaconic acid (IA), a C5-dicarboxylic acid, is a potential bio-based building block for the polymer industry. There are three pathways for IA production from natural IA producers; however, most of the engineered strains were used for IA production by heterologous expression of cis-aconitate decarboxylase gene (cadA) from Aspergillus terreus. In this study, IA was produced by an engineered Corynebacterium glutamicum ATCC 13032 expressing two different types of genes from two distinct pathways. The first involves the mammalian immunoresponsive gene1 (Irg1) derived from Mus musculus. The second (termed here the trans-pathway) involves two genes from the natural IA producer Ustilago maydis which are aconitate-delta-isomerase (Adi1) and trans-aconitate decarboxylase (Tad1) genes. The constructed strains developing the two distinct IA production pathways: C. glutamicum ATCC 13032 pCH-Irg1opt and C. glutamicum ATCC 13032 pCH-Tad1optadi1opt were used for production of IA from different carbon sources. The results reflect the possibility for IA production from C. glutamicum expressing the trans-pathway (Adi1/Tad1 genes) and cis-pathway (Irg1 gene) other than the well-known cis-pathway that depends mainly on cadA gene from A. terreus. The developed strain expressing trans-pathway from U. maydis; however, proved to be better at IA production with high titers of 12.25, 11.34, and 11.02 g/L, and a molar yield of 0.22, 0.42, and 0.43 mol/mol from glucose, maltose, and sucrose, respectively, via fed-batch fermentation. The present study suggests that trans-pathway is better than cis-pathway for IA production in engineered C. glutamicum.


Using artificial intelligence in a primary care setting to identify patients at risk for cancer: a risk prediction model based on routine laboratory tests.
To evaluate the ability of an artificial intelligence (AI) model to predict the risk of cancer in patients referred from primary care based on routine blood tests. Results obtained with the AI model are compared to results based on logistic regression (LR). An analytical profile consisting of 25 predefined routine laboratory blood tests was introduced to general practitioners (GPs) to be used for patients with non-specific symptoms, as an additional tool to identify individuals at increased risk of cancer. Consecutive analytical profiles ordered by GPs from November 29th 2011 until March 1st 2020 were included. AI and LR analysis were performed on data from 6,592 analytical profiles for their ability to detect cancer. Cohort I for model development included 5,224 analytical profiles ordered by GP's from November 29th 2011 until the December 31st 2018, while 1,368 analytical profiles included from January 1st 2019 until March 1st 2020 constituted the "out of time" validation test Cohort II. The main outcome measure was a cancer diagnosis within 90 days. The AI model based on routine laboratory blood tests can provide an easy-to use risk score to predict cancer within 90 days. Results obtained with the AI model were comparable to results from the LR model. In the internal validation Cohort IB, the AI model provided slightly better results than the LR analysis both in terms of the area under the receiver operating characteristics curve (AUC) and PPV, sensitivity/specificity while in the "out of time" validation test Cohort II, the obtained results were comparable. The AI risk score may be a valuable tool in the clinical decision-making. The score should be further validated to determine its applicability in other populations.


Worsening of kidney function in patients with atrial fibrillation and chronic kidney disease: evidence from the real-world CALLIPER study.
Evidence is needed on the impact of anticoagulation therapy on kidney function in patients with atrial fibrillation (AF). The objective of this analysis, which is part of the CALLIPER study, was to investigate the risk of worsening kidney function with rivaroxaban 15 mg once daily compared with warfarin in patients with AF and moderate-to-severe chronic kidney disease (CKD) in routine clinical practice in the United States. CALLIPER was an observational, retrospective, new-user cohort study. Adult patients with AF in the US IBM Watson MarketScan databases who newly initiated anticoagulation with rivaroxaban 15 mg once daily or warfarin between January 2013 and December 2017 were included. Comparative analysis was performed using Cox proportional hazards regression after adjustment for potential confounding by the stabilized inverse probability of treatment weighting approach and propensity score matching. One of the main study outcomes was worsening kidney function (composite of progression to CKD stage 5, kidney failure, or need for dialysis), besides traditional AF-related outcomes. The cohort included 7368 patients: 5903 (80.1%) initiating warfarin and 1465 (19.9%) initiating rivaroxaban 15 mg once daily. Rivaroxaban 15 mg was associated with a significant 47% reduction in the risk of worsening kidney function versus warfarin (hazard ratio 0.53; 95% confidence interval 0.35-0.78). Similar results were observed in the subgroup of patients with type 2 diabetes. Rivaroxaban 15 mg may be associated with a lower risk of worsening kidney function as compared with warfarin in the atrial fibrillation population with moderate-to-severe CKD. NCT03359876.


Watershed-scale impacts of stormwater green infrastructure on hydrology, nutrient fluxes, and combined sewer overflows in the mid-Atlantic region.
Stormwater green infrastructure (SGI), including rain gardens, detention ponds, bioswales, and green roofs, is being implemented in cities across the globe to reduce flooding, combined sewer overflows, and pollutant transport to streams and rivers. Despite the increasing use of urban SGI, few studies have quantified the cumulative effects of multiple SGI projects on hydrology and water quality at the watershed scale. To assess the effects of SGI, Washington, DC, Montgomery County, MD, and Baltimore County, MD, were selected based on the availability of data on SGI, water quality, and stream flow. The cumulative impact of SGI was evaluated over space and time by comparing watersheds with and without SGI, and by assessing how long-term changes in SGI impact hydrologic and water quality metrics over time. Most Mid-Atlantic municipalities have a goal of achieving 10-20% of the landscape drain runoff through SGI by 2030. Of these areas, Washington, DC currently has the greatest amount of SGI (12.7% of the landscape drained through SGI), while Baltimore County has the lowest (7.9%). When controlling for watersheds size and percent impervious surface cover, watersheds with greater amounts of SGI have less flashy hydrology, with 44% lower peak runoff, 26% less frequent runoff events, and 26% less variable runoff. Watersheds with more SGI also show 44% less NO3(-) and 48% less total nitrogen exports compared to watersheds with minimal SGI. There was no significant reduction in phosphorus exports or combined sewer overflows in watersheds with greater SGI. When comparing individual watersheds over time, increases in SGI corresponded to non-significant reductions in hydrologic flashiness compared to watersheds with no change in SGI. While the implementation of SGI is somewhat in its infancy in some regions, cities are beginning to have a scale of SGI where there are statistically significant differences in hydrologic patterns and water quality.


Biological and biochemical characterisation of purine analogue resistant clones of V79 Chinese hamster cells.
Purine analogue resistant clones have been selected from the closely related Chinese hamster lines V79A and V79S. Clones were of either spontaneous origin or induced by EMS or ultraviolet light. The majority of clones selected in 8-azaguanine showed stable cross resistance to 6-thioguanine. Clones derived from V79A and selected for 6-thioguanine resistance were cross resistant to 8-azaguanine: however a group of 6-thioguanine resistant mutants selected from V79S cells were 8-azaguanine sensitive. All clones except two were unable to grow in HAT medium. The two exceptions were 8-azaguanine resistant, showed partial sensitivity to 6-thioguanine, and also differed in other biochemical characteristics. HGPRT activity was measurable in extracts of all clones under standard conditions. In many clones, HGPRT activity increased as the hypoxanthine concentration was reduced. Whole cell uptake of [14C] hypoxanthine was low in all cases examined and was not modified by incubation in the presence of amethopterin. The heat sensitivity and electrophoretic mobility of HGPRT in extracts of some clones was compared to that in wild-type extracts. All clones tested except one, which was consistently HAT positive, showed enhanced heat sensitivity and reduced electrophoretic mobility. None of the mutants reverted spontaneously at detectable frequency but some could be induced to revert by EMS. The presence of measurable enzyme with altered properties in all clones suggests that these revertable drug resistant clones represent missense mutants.


PyLiger: scalable single-cell multi-omic data integration in Python.
LIGER (Linked Inference of Genomic Experimental Relationships) is a widely used R package for single-cell multi-omic data integration. However, many users prefer to analyze their single-cell datasets in Python, which offers an attractive syntax and highly optimized scientific computing libraries for increased efficiency. We developed PyLiger, a Python package for integrating single-cell multi-omic datasets. PyLiger offers faster performance than the previous R implementation (2-5× speedup), interoperability with AnnData format, flexible on-disk or in-memory analysis capability and new functionality for gene ontology enrichment analysis. The on-disk capability enables analysis of arbitrarily large single-cell datasets using fixed memory. PyLiger is available on Github at https://github.com/welch-lab/pyliger and on the Python Package Index. Supplementary data are available at Bioinformatics online.


Reduced Myelin Water in the White Matter Tracts of Patients with Niemann-Pick Disease Type C.
Previous studies using diffusion tensor imaging to examine white matter in Niemann-Pick disease type C have produced mixed results. However, diffusion tensor imaging does not directly measure myelin and may be affected by other structural changes. We used myelin water imaging to more directly examine demyelination in 2 patients with Niemann-Pick disease type C. The results suggest that this technique may be useful for identifying regional changes in myelination in this condition.


Identification of subcutaneous myiasis using bedside emergency physician performed ultrasound.
Subcutaneous myiasis, a maggot infiltration of human tissue, is common in tropical countries. However, physicians in the United States may be unlikely to consider this etiology of dermatologic abnormalities even when a travel history suggests the diagnosis should be included in the differential. We report the case of a patient who returned from Sierra Leone with an infestation of a maggot of Cordylobia anthropophaga (tumbu fly) that was diagnosed and appropriately treated based on ultrasound findings. As international travel increases, clinicians should maintain a high level of suspicion for tumbu fly infestation in returned travelers from endemic areas. The increasing use of ultrasound in the Emergency Department for evaluation of skin and soft tissue infections may aid the physician in making the diagnosis of subcutaneous myiasis.


Moyamoya disease: current concepts and future perspectives.
Moyamoya disease is an uncommon cerebrovascular disease that is characterised by progressive stenosis of the terminal portion of the internal carotid artery and its main branches. The disease is associated with the development of dilated, fragile collateral vessels at the base of the brain, which are termed moyamoya vessels. The incidence of moyamoya disease is high in east Asia, and familial forms account for about 15% of patients with this disease. Moyamoya disease has several unique clinical features, which include two peaks of age distribution at 5 years and at about 40 years. Most paediatric patients have ischaemic attacks, whereas adult patients can have ischaemic attacks, intracranial bleeding, or both. Extracranial-intracranial arterial bypass, including anastomosis of the superficial temporal artery to the middle cerebral artery and indirect bypass, can help prevent further ischaemic attacks, although the beneficial effect on haemorrhagic stroke is still not clear. In this Review, we summarise the epidemiology, aetiology, clinical features, diagnosis, surgical treatment, and outcomes of moyamoya disease. Recent updates and future perspectives for moyamoya disease will also be discussed.


Recombinant and endogenous ways to produce methylated phospholipids in Escherichia coli.
Escherichia coli is the daily workhorse in molecular biology research labs and an important platform microorganism in white biotechnology. Its cytoplasmic membrane is primarily composed of the phospholipids phosphatidylethanolamine (PE), phosphatidylglycerol (PG), and cardiolipin (CL). As in most other bacteria, the typical eukaryotic phosphatidylcholine (PC) is not a regular component of the E. coli membrane. PC is known to act as a substrate in various metabolic or catabolic reactions, to affect protein folding and membrane insertion, and to activate proteins that originate from eukaryotic environments. Options to manipulate the E. coli membrane to include non-native lipids such as PC might make it an even more powerful and versatile tool for biotechnology and protein biochemistry. This article outlines different strategies how E. coli can be engineered to produce PC and other methylated PE derivatives. Several of these approaches rely on the ectopic expression of genes from natural PC-producing organisms. These include PC synthases, lysolipid acyltransferases, and several phospholipid N-methyltransferases with diverse substrate and product preferences. In addition, we show that E. coli has the capacity to produce PC by its own enzyme repertoire provided that appropriate precursors are supplied. Screening of the E. coli Keio knockout collection revealed the lysophospholipid transporter LplT to be responsible for the uptake of lyso-PC, which is then further acylated to PC by the acyltransferase-acyl carrier protein synthetase Aas. Overall, our study shows that the membrane composition of the most routinely used model bacterium can readily be tailored on demand.Key points• Escherichia coli can be engineered to produce non-native methylated PE derivatives.• These lipids can be produced by foreign and endogenous proteins.• Modification of E. coli membrane offers potential for biotechnology and research.


Isolation and characterization of amphotericin B-resistant cell lines in Chinese hamster cells.
Amphotericin B is a polyene macrolide antibiotic which interacts specifically with sterols in mammalian cell membranes. Amphotericin B-resistant (AMBr) lines of stable phenotype have been isolated from cultured Chinese hamster (V79) cells. Three AMBr clones (AMBr-1, -2 and -3) isolated independently after treatment with nitrosoguanidine were resistant to greater than or equal to microgram/ml of the antibiotic, while DNA synthesis as well as the colony-forming ability of the parental V79 cells was blocked by greater than 80% of control in the presence of 20--50 microgram/ml amphotericin B. The AMBr cell line also exhibited increased resistance to other polyene macrolide antibiotics such as nystatin and pentamycin. Other agents, however, such as cytosine arabinoside or ricin, blocked DNA synthesis in AMBr cells to the same extent as in V79 cells. The amphotericin B resistance phenotype was stably retained even after AMBr cells were cultured in the absence of the drug for over 200 generations. The control of free cholesterol or its esters was significantly decreased in all three resistant clones. Furthermore, cholesterol synthesis from acetate as well as mevalonate was partly defective in AMBr cells, compared with that in V79 cells.


circ_2858 Helps Blood-Brain Barrier Disruption by Increasing VEGFA via Sponging miR-93-5p during Escherichia coli Meningitis.
Meningitic Escherichia coli invasion of the host brain can lead to increased blood-brain barrier (BBB) permeability. Circular RNAs (circRNAs) are non-coding RNAs, highly abundant in the brain, that are widely involved in the pathological processes of central nervous system (CNS) disorders; however, whether circRNAs participate in the regulation of BBB permeability during E. coli meningitis remains unknown. Here, we identified a novel circRNA, circ_2858, that was significantly upregulated in human brain microvascular endothelial cells (hBMECs) upon meningitic E. coli infection. We also found that circ_2858 regulated BBB permeability in hBMECs by competitively binding miR-93-5p, thereby inducing the upregulation of vascular endothelial growth factor A and finally resulting in downregulation as well as altered distribution of tight junction proteins such as ZO-1, Occludin, and Claudin-5. These findings provide novel insights into the influence of circ_2858 on BBB permeability during the pathogenic process of E. coli meningitis, suggesting potential nucleic acid targets for future prevention and therapy of CNS infection induced by meningitic E. coli.


Malaria Is Associated with Diminished Levels of Ascorbic Acid: A Systematic Review and Meta-Analysis.
Background: It is still unclear how ascorbic acid levels relate to the pathogenesis of malaria. This systematic review synthesized different ascorbic acid levels in malaria patients with different severity levels of malaria and Plasmodium species. Methods: The systematic review protocol was registered in the PROSPERO database (CRD42023394849). A systematic search of PubMed, Embase, MEDLINE, Ovid, Scopus, and Google Scholar was conducted to identify studies that reported ascorbic acid and malaria. The pooled standardized mean difference (Cohen's d) with 95% confidence intervals (CIs) was calculated using the random-effects model. Results: A total of 1480 articles were obtained from the searches of the databases, and 30 studies were included for syntheses. The meta-analysis revealed that patients with malaria had lower levels of ascorbic acid than those without malaria or uninfected controls (p < 0.01, Cohen's d = -3.71, 95% CI = -4.44 to -2.98, I2 = 98.87%, 30 studies). Comparable levels of ascorbic acid were observed between patients with severe malaria and those with nonsevere malaria (p = 0.06, Cohen's d = -1.39, 95% CI = -2.85 to 0.07, I2 = 96.58%, 4 studies). Similarly, levels of ascorbic acid were comparable between patients with Plasmodium falciparum and Plasmodium vivax malaria (p = 0.34, Cohen's d = -1.06, 95% CI = -3.23 to 1.12, I2 = 97.30%, 3 studies). Conclusions: The meta-analysis reveals diminished levels of ascorbic acid in malaria cases. Manipulating the host's nutritional status, such as by supplementing it with ascorbic acid to restore reactive oxygen species balance, may alter the progression of malarial infection and prevention of disease severity.


Effect of antiresorptive therapy on aromatase inhibitor induced bone loss in postmenopausal women with early-stage breast cancer: A systematic review and meta-analysis of randomized controlled trials.
Aromatase inhibitors (AIs) are routinely used to treat postmenopausal women with early-stage breast cancer. Although AIs improve breast cancer outcomes, they increase the risk of osteoporosis and fractures. This systematic review and meta-analysis assesses the effect of antiresorptive drugs on AI induced bone loss in postmenopausal women with non-metastatic breast cancer. We searched four databases until November 4th 2020. We included Randomized controlled trials (RCTs) of antiresorptive drugs in postmenopausal women with breast cancer treated with AI. Two authors screened studies, extracted data and assessed the risk of bias independently and in duplicate. We identified 14 RCTs: 7 on zoledronic acid, 6 on oral bisphosphonates and 1 on denosumab. The mean difference in bone mineral density (BMD) was 5% at the lumbar spine and 4% at the total hip, at 12 months, favoring zoledronic acid compared to control. The certainty of the evidence was low for lumbar spine and moderate for total hip BMD. Similarly, the mean difference was 3% at the lumbar spine and 2% at the total hip, favoring oral bisphosphonates with moderate certainty. The mean difference was 6% at the lumbar spine, and 4% at the total hip BMD favoring denosumab compared to placebo. In addition, zoledronic acid resulted in a mean difference in bone turnover marker levels of -35-41%, and the relarive risk for morphometric vertebral fractures was 0.7 [0.3-1.4], compared to control. Denosumab reduced fracture incidence by 50% compared to placebo. Evidence suggests a protective effect of antiresorptive drugs on BMD and bone turnover markers in postmenopausal women with non-metastatic breast cancer on AI. However, data on fracture risk reduction remains unclear.


The possible significance of the baropathic nature of keratectasias.
The aim was to describe the baropathic nature of the keratectasias and to examine the possible significance of intraocular pressure-elevating activities in the development and/or progression of these conditions. Articles were selected from 150 produced from a PubMed search for keratectasias and used to elucidate the biomechanics and dependence on intraocular pressure of those conditions. The combination of viscoelastic and baropathic features of keratectasia indicate that elevations in intraocular pressure have the potential to contribute to the development and/or progression of these conditions. Acute events such as hydrops and perforation appear to be more or less dependent on IOP elevation. Development and/or progression of keratectasia might be slowed by patient counselling, which explains the dependence of keratectasia on intraocular pressure and recommends avoidance and/or moderation of activities that elevate intraocular pressure. Successful adoption of such advice could reduce rates of disease progression and the need for refitting with more expensive contact lens designs, which are often required for advanced cases, as well as reduce the need for grafting and the drain on limited availability of donor corneas.


Procalcitonin and neopterin as indicators of infection in critically ill patients.
: In critically ill patients, severe infection and systemic inflammation due to non-infectious causes produce very similar clinical presentations, and traditional infection markers do not always differentiate these two conditions. Both procalcitonin and neopterin have been suggested to aid in the early diagnosis of bacterial infections and in differentiating bacterial infections from systemic inflammatory, non-infectious diseases or from viral infections. : Procalcitonin (PCT) and neopterin were analyzed in 208 ICU patients who developed acute fever or septic shock. Blood samples were taken every 8th h within 48 h of the onset of fever or septic shock. : A total 162/208 of patients had infection, the most common location being the respiratory tract. Mortality was higher in infected patients (31.4% vs. 10.9%; P < 0.01). The optimum cut-off levels in identifying patients with infection of daily peak PCT were 0.8 microg/L on day 1 and 0.9 microg/L on day 2, and both sensitivity (67.7% and 60.9%, respectively) and specificity (47.8% and 63%) were poor. Accordingly, the optimum cut-off values of peak neopterin were 18 and 16 pg/L. The sensitivity was 62.7% on day 1 and 69.3% on day 2, while specificity was correspondingly 78.3% and 67.9%. There were no significant differences between the markers in discriminating between patients with infection or inflammation. Both PCT and neopterin increased with the severity of infection. They were higher in non-survivors. : PCT and neopterin were equally effective, although not very accurate in differentiating between infection and inflammation in critically ill patients. Neopterin was more specific than PCT, suggesting that neopterin is related to the activity of inflammatory response.


Ibrutinib Monotherapy in Relapsed or Refractory, Transformed Diffuse Large B-cell Lymphoma.
Histologic transformation to diffuse large B-cell lymphoma (tDLBCL) occurs in a significant proportion of indolent lymphomas. However, few studies of novel agents inform its management, particularly when relapsed after or refractory (R/R) to prior treatment. We prospectively evaluated ibrutinib monotherapy in pathologically documented patients with R/R tDLBCL in a single-arm study. The primary endpoint was overall response rate. Twenty patients who had received a median of 4 (range, 2-9) prior lines of therapy overall (median, 2.5; range, 1-9 for tDLBCL) were treated. The overall response rate was 35%, including complete responses in 15%. The median progression-free survival and overall survival were 4.1 months (95% confidence interval, 2.4-6.2 months) and 22.4 months (95% confidence interval, 7.5 months to not reached), respectively. Disease control > 2 months was seen in 75% and > 1 year in 15%. Response was associated with either low tumor bulk or low metabolic tumor volume (P = .05) but not with antecedent lymphoma histology (P = 1.0). Treatment-related adverse events were consistent with prior studies of ibrutinib. Ibrutinib showed low toxicity and meaningful efficacy in R/R tDLBCL, including short-term disease control in most cases. Results demonstrate the potential utility of ibrutinib in this challenging clinical setting, including as a potential bridge to more definitive treatments.


Onchocerca armillata contains the endosymbiotic bacterium Wolbachia and elicits a limited inflammatory response.
Human onchocerciasis, also known as River Blindness, is a debilitating disease caused by the filarial nematode Onchocerca volvulus. Many, but not all, filarial nematodes carry within their tissues endosymbiotic, Rickettsia-like bacteria of the genus Wolbachia. Onchocerca spp. infections in cattle offer the most relevant, analogous host-parasite model system. West African cattle are commonly co-infected with four Onchocerca spp.; two of these are Wolbachia-positive (Onchocerca gutturosa and Onchocerca ochengi), and the remainder are of unknown Wolbachia status (Onchocerca dukei and Onchocerca armillata). Previous studies have suggested that worm survival is dependent on this bacterium. O. armillata, an abundant parasite of African cattle that has received little attention, is a primitive species that may lack Wolbachia. The objectives of this study were to determine if O. armillata carries Wolbachia and to provide preliminary descriptions of the host inflammatory cell environment around the adult worms. The findings may support or refute the hypothesis that a prime contribution of Wolbachia is to permit long-term survival and reproduction of certain Onchocerca spp. (including O. volvulus in humans). O. armillata adult worms were found in the aorta of 90.7% of cattle (n=54) slaughtered at an abattoir in Ngaoundéré, Adamawa Region, Cameroon. The presence of Wolbachia in O. armillata was confirmed by a specific anti-Wolbachia surface protein antibody detected using a peroxidase conjugate (immunohistochemistry) and PCR for detection of Wolbachia-specific sequences within DNA extracts from frozen worms. Tissue sections stained with haematoxylin and eosin showed the host cell response to be dominated by macrophages and fibroblasts. This is unusual compared with nodule-dwelling Wolbachia-positive Onchocerca spp., where the host response is typically characterised by granulocytes, and suggests that the mechanisms for worm survival employed by this species (which is probably motile) may differ.


The elements of community consultation.
Psychiatrists are involved in both clinical and institutional consultations. Although the tradition of psychiatric consultation for medical patients is well established and adequately described, there are few contributions concerning the conceptual framework for community consultations and few case examples. In order to provide a framework for conceptualizing consultation work this communication presents an example of a consultation to a community agency and discusses some of the specific elements in community consultation. Thus it may help professionals to practice the science and art of consultation to community agencies.


Synthesis and antibacterial activity of novel 3-O-arylalkylcarbamoyl-3-O-descladinosyl-9-O-(2-chlorobenzyl)oxime clarithromycin derivatives.
A novel series of 3-O-arylalkylcarbamoyl-3-O-descladinosyl-9-O-(2-chlorobenzyl)oxime clarithromycin derivatives, were designed, synthesized and evaluated for their in vitro antibacterial activity. These derivatives were found to have strong activity against susceptible and resistant bacteria strains. Among them, compounds 7a and 7q showed the most potent activity (0.125 µg/mL) against erythromycin-resistant S. pneumoniae expressing the mefA gene. Moreover, compounds 7f, 7i, 7p and 7z displayed remarkably improved activity (4 µg/mL) against penicillin-resistant S. aureus ATCC31007, and compounds 7a, 7b, 7f, 7p and 7z showed improved activity (8 µg/mL) against erythromycin-resistant S. pyogenes. In particular, compound 7z exhibited potent and balanced activity against the tested drug-susceptible and -resistant bacterial strains.


Midgut volvulus in the adult. A complication of intestinal malrotation.
We report two cases of abnormalities in the second and third stages of intestinal rotation and fixation which presented in adulthood as recurrent midgut volvulus. We review the embryology, symptoms, diagnosis, and treatment of this problem.


Cerebellopontine Angle Lipoma with Contralateral Sudden Deafness: A Case Report and Review of Literature.
Cerebellopontine angle lipoma is a rare tumor that composes less than 1% of all CPA tumors. There has been no recorded case of unilateral CPA/IAC lipoma with sudden contralateral deafness yet. We report a 52-year-old man diagnosed with right cerebellopontine angle lipoma and combined left total deafness. Pure-tone audiometry revealed total sensorineural deafness in his left ear and moderate sensorineural deafness in the right ear. The patient was treated with glucocorticoids, batroxobin, and other symptomatic treatments. There was no substantial improvement in hearing after 14 days' treatment. We chose conservative treatment for him. It is advised to wear hearing aids in the right ear and to undergo regular imaging monitoring. Treatment options for such patients should be chosen by taking into account the degree of bilateral hearing loss, the size and location of the tumor, the possibility of preserving hearing during surgery, the functional level of the patient's facial nerve, and other factors.


Sulfamethoxazole:hydroxypropyl-β-cyclodextrin complex: preparation and characterization.
A complex of sulfamethoxazole (SMX) and hydroxypropyl-β-cyclodextrin (HP-β-CD) was developed and characterized in order to investigate their interactions in aqueous solution and the solid state. The SMX solubility was significantly increased upon complexation with HP-β-CD, with the solubility isotherm being an A(N) type due to the presence of aggregates and the stability constant calculated for a 1:1 complex being 302 ± 3 M⁻¹. Fourier-transform infrared (FT-IR) spectroscopy and scanning electron microscopy (SEM) experiments were used to compare the freeze-dried system with a physical mixture, and demonstrated the complex formation in the solid state. The differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA) showed that the thermal stability of SMX was enhanced in the presence of HP-β-CD.


Use of direct gradient analysis to uncover biological hypotheses in 16s survey data and beyond.
This study investigated the use of direct gradient analysis of bacterial 16S pyrosequencing surveys to identify relevant bacterial community signals in the midst of a "noisy" background, and to facilitate hypothesis-testing both within and beyond the realm of ecological surveys. The results, utilizing 3 different real world data sets, demonstrate the utility of adding direct gradient analysis to any analysis that draws conclusions from indirect methods such as Principal Component Analysis (PCA) and Principal Coordinates Analysis (PCoA). Direct gradient analysis produces testable models, and can identify significant patterns in the midst of noisy data. Additionally, we demonstrate that direct gradient analysis can be used with other kinds of multivariate data sets, such as flow cytometric data, to identify differentially expressed populations. The results of this study demonstrate the utility of direct gradient analysis in microbial ecology and in other areas of research where large multivariate data sets are involved.


alpha-Tocopherol disturbs macrophage LXRalpha regulation of ABCA1/G1 and cholesterol handling.
Based on the oxidation hypothesis high doses of alpha-tocopherol have been advocated to prevent atherosclerosis, but clinical trials failed to demonstrate a benefit. As specific oxylipids activate PPARgamma and LXRalpha, master regulators of lipid metabolism and cholesterol exporters, we hypothesized, that high dose alpha-tocopherol might interfere with reverse cholesterol transport out of the vessel wall. Human THP-1 cells, a foam cell model, were preincubated with alpha-tocopherol or carrier before exposure to oxidized LDL, delipidated HDL or control buffer. Specific mRNAs were quantified by real-time RT-PCR, LXRalpha activation by a reporter gene assay and cellular cholesterol homeostasis by oxLDL and dHDL facilitated uptake and efflux assays. alpha-Tocopherol significantly reduced baseline expression and stimulation by oxLDL of LXRalpha activity, CD36, ABCA1, and ABCG1. alpha-Tocopherol also reversed the suppression of CD36 and ABCA1 by dHDL. Thus alpha-Tocopherol compromises cellular lipid scavenging and channelling of cholesterol into reverse transport out of the vessel wall.


Purification and characterization of thiol aminopeptidase from the cytosolic fraction of human placenta.
A thiol-dependent aminopeptidase was purified from the cytosolic fraction of human placenta. The purified enzyme consisted of a single polypeptide chain with a mol wt of 95,000. The enzyme was most active in the neutral region with Ala-pNA as substrate, and the activity was increased about 20-fold in the presence of some -SH compounds. The results of substrate specificity studies indicated that the enzyme hydrolyzes bonds involving the amino groups of neutral and basic amino acid residues. However, higher thiol-dependent activity was only detected with neutral ones. The enzyme was strongly inhibited by microbial aminopeptidase inhibitors, puromycin, o-phenanthroline, and sulfhydryl reactive-reagents. As to several naturally occurring peptides tested, the enzyme showed N-terminal Tyr-releasing activity toward enkephalins and kinin-converting activity.


Highly selective oxidation of organic contaminants in the RuIII-activated peroxymonosulfate process: The dominance of RuVO species.
Ruthenium (RuIII)-activated peroxymonosulfate (the RuIII/PMS process) is one of the most efficient PMS-based advanced oxidation processes for the abatement of organic contaminants. Here we interestingly found that phenyl methyl sulfoxide (PMSO) was significantly oxidized to phenyl methyl sulfone (PMSO2) in the RuIII/PMS process at the pH range of 3.0-6.0, with the conversion ratio of ΔPMSO to ΔPMSO2 was close to 100%, which favored the dominance of high-valent ruthenium-oxo species (RuVO) instead of the widely-recognized radicals (i.e, hydroxyl radical and sulfate radical). Scavenging experiments further indicated that RuVO was unreactive to tert-butyl alcohol, but could be scavenged by methanol and dimethyl sulfoxide. Besides, sulfamethoxazole, acetaminophen, carbamazepine, diclofenac, 2,4,6-trichlorophenol were readily degraded in the RuIII/PMS process, but atrazine, ibuprofen, benzoic acid and 4-nitrobenzoic acid were barely removed, suggesting the high selectivity of RuVO species. This study enriched the understandings on the mechanism of RuIII-mediated PMS activation and the nature of RuVO species.


Immunopathogenesis of simian immunodeficiency virus infection in nonhuman primates.
Soon after the discovery of HIV-infected humans, rhesus macaques in a colony at the New England Primate Research Center showed similar signs of a progressive immune suppression. The discovery of the simian immunodeficiency virus (SIV)-associated disease opened the door to study an AIDS-like illness in nonhuman primates (NHP). Even after 3 decades, this animal model remains an invaluable tool to provide a greater insight into HIV immunopathogenesis. In this review, recent progress in deciphering pathways of immunopathogenesis in SIV-infected NHP is discussed. The immense diversity of mutations in SIV stocks prepared at different laboratories has recently been realized. The massive expansion of the enteric virome is a key finding in SIV-induced immunopathogenesis. Defining the function of host restriction factors, like the recently discovered SAMHD1, helps to evaluate the impact of the innate immune responses on virus replication. Utilization of pyrosequencing and defining molecular mechanisms of major histocompatibility complex (MHC) class I restriction helps to understand how the virus evades CD8 T-cell responses. The definition of MHC class I molecules in different NHP species provides new animal models to study SIV immunopathogenesis. T follicular helper cells have gained major interest in characterizing humoral immune responses in SIV infection and AIDS vaccine strategies. The ability of natural hosts to remain disease-free despite ongoing replication of SIV is continuing to puzzle the field. The HIV research field continues to realize the immense complexity of the host virus interaction. NHP present an invaluable tool to make progress towards an effective AIDS vaccine.


Truly-Biocompatible Gold Catalysis Enables Vivo-Orthogonal Intra-CNS Release of Anxiolytics.
Being recognized as the best-tolerated of all metals, the catalytic potential of gold (Au) has thus far been hindered by the ubiquitous presence of thiols in organisms. Herein we report the development of a truly-catalytic Au-polymer composite by assembling ultrasmall Au-nanoparticles at the protein-repelling outer layer of a co-polymer scaffold via electrostatic loading. Illustrating the in vivo-compatibility of the novel catalysts, we show their capacity to uncage the anxiolytic agent fluoxetine at the central nervous system (CNS) of developing zebrafish, influencing their swim pattern. This bioorthogonal strategy has enabled -for the first time- modification of cognitive activity by releasing a neuroactive agent directly in the brain of an animal.


Immunobiological biochemical and physico-chemical characteristics of Brucella lipopolysaccharide subjected to various doses of gamma radiation.
A comparative study of toxicity, serological activity, some biochemical and physico-chemical properties of the highly toxic Brucella lipopolysaccharide (LPS) and of preparations obtained after the action of gamma radiation in doses of 1.3 and 10 mrad on the antigen. The toxicity of LPS was found to decrease with the increasing dose of irradiation. Irradiation with a dose of 3 mrad produced a marked decrease in the toxicity of the antigen without causing essential changes in its serological properties. The process of LPS detoxication under the effect of irradiation was accompanied by changes in certain biochemical and physico-chemical indices suggestive of the modification of the primary structure of the LPS molecule and of impairment of mainly its polysaccharide side chains.


Synthesis of a bile acid-based click-macrocycle and its application in selective recognition of chloride ion.
A novel method for the synthesis of a bile acid-based macrocycle has been developed using click chemistry. The 1,2,3-triazolium derivative of the macrocycle shows remarkable selectivity in binding of chloride ion.


Colony forming ability of human breast carcinomas: lack of prognostic significance.
To study whether colony growth in vitro reflects the prognosis of breast cancer patients, specimens from a total number of 138 patients with primary breast carcinomas were cultivated in the Courtenay-Mills soft agar method. The plating efficiency (PE) values were related to various clinical and histopathological parameters. No significant correlation was found between colony forming ability and menopausal status, histopathology, TNM-status or steroid hormone receptor status. The crude survival of the patients was not significantly correlated to the in vitro growth of the tumours; neither was there any difference in relapse-free survival between patients whose tumours failed to grow in vitro and those having growing tumours (PE greater than 0). A multivariate survival analysis of 115 patients with primary tumours without distant metastases revealed that the PE was not a significant independent prognostic indicator, as it gave no additional prognostic information above that of node and ER status. It is concluded that routine measurement of colony formation in vitro is not warranted in the management of breast cancer.


Preventing staple-line leak in sleeve gastrectomy: reinforcement with bovine pericardium vs. oversewing.
One of the most serious, potentially life-threatening complications of laparoscopic sleeve gastrectomy (LSG) is staple-line leakage. Oversewing the LSG staple line vs buttressing it with bovine pericardial strips (BPS) to reduce perioperative bleeding and postoperative gastric leak was evaluated. From 2006 through 2011, 160 patients underwent LSG with suturing as the only staple-line reinforcement (Group A). From March 2010 through August 2012, 84 LSG patients had BPS incorporated into their last two stapler firings (Group B). Staple lines were evaluated perioperatively for bleeding, and patients were monitored for indications of staple-line leaks (peritonitis, abnormal output from the drain). In preoperative Group A and B, there were 117 (73.1%) vs. 56 (66.7%) females; mean age, 35.2 years (18.0-68.0) vs. 33.8 years (15.0-64.0); mean body mass index (BMI, kilograms per square meter), 42.5 (27.0-76.0) vs. 42.0 (30.0-58.0). Three months after surgery, mean BMI for Group A was 37.3 (-5.9); Group B, 35.2 (-7.3); at 6 months, 32.7 (-10.8) and 31.5 (-11.3; p < 0.001). Although there was no significant difference in perioperative blood loss, oversewn staple lines in Group A often required electrocautery to stanch bleeding; this was not required for Group B. In Group A, 15 patients (9.4%) developed complications; in Group B, five (6.0%; p = 0.46). Gastric fistula, verified by barium swallow, occurred in eight Group A patients (5.0%); in Group B, one (1.2%; p = 0.17). Relative to oversewing, staple-line buttressing with bovine pericardium was readily accomplished, safe, and associated with a lower staple-line leak rate.


Widening Racial Disparities During COVID-19 Telemedicine Transition: A Study of Child Mental Health Services at Two Large Children's Hospitals.
To examine whether racial disparities in access to pediatric mental health care were affected during the COVID-19 telemedicine transition at both The Children's Hospital of Philadelphia (CHOP) and Boston Children's Hospital (BCH). Electronic health records were queried for all unique outpatient visits from a pre-pandemic period in 2019 and a within-pandemic period in 2020. Changes in the proportion of patients were compared based on insurance status, clinic location, and racial identification. Hypotheses were tested via logistic regression analyses. At CHOP, from 2019 to 2020, the proportion of racially minoritized patients significantly declined within a 1-month period from 62% to 51%, whereas the proportion of White-identifying patients increased from 38% to 49% (β = 0.47; z = 3.60; p =.0003), after controlling for insurance status and clinic location. At BCH, the proportion of racially minoritized patients significantly declined within a longer 6-month period between 2019 and 2020, from 62% to 59%, whereas the proportion of White-identifying patients increased from 38% to 41% (β = 0.13; z = 2.8; p = .006), after controlling for insurance status. At CHOP and BCH, the COVID-19 telemedicine transition exacerbated pre-existing racial disparities in pediatric mental health services. Our findings suggest that racially minoritized patients receiving services in urban areas may be particularly at risk for losing access when telemedicine is implemented. Although there are limitations to this racial dichotomization, examining differences between White and racially minoritized patients can highlight ways in which White-identifying individuals have disproportionately received enhanced access to healthcare resources.


Linking circular economy model and green supply chain practices to shape environmental performance: case of Vietnam.
The circular economy (SE) is the foremost element for environmental improvement that reduces wastage and enhances production. This aspect requires the focus of new researchers and experienced policymakers. Hence, the current research examines the impact of SE practices such as internal environmental management (IEM), eco-design, and internal recovery on the SE targeted performance, such as environmental performance in Vietnam. The research also investigates the moderating role of environmental-oriented supply chain (SC) practices such as green purchases among IEM, eco-design, internal recovery, and environmental performance in Vietnam. The research collected the data from the respondents using survey questionnaires. The research also investigates the association among variables using smart-PLS. The outcomes revealed that the IEM, eco-design, and internal recovery are positively associated with environmental performance. The outcomes also exposed that the green purchases significantly moderates among IEM, eco-design, internal recovery, and environmental performance. The research guides the regulators in establishing regulations related to achieve the high SE targeted performance, such as environmental performance using circular economy practices such as IEM, eco-design, and internal recovery.


A field evaluation of anthelmintics in horses in Sweden.
A field evaluation of anthelmintics in 336 horses on 37 farms was conducted between February and May 1986 in Sweden. The herds, each comprising at least eight horses, had histories of grazing on permanent pastures and receiving regular treatments against parasites at least three times a year. Small strongyles were refractory to (pro)-benzimidazole drugs in all but one of 23 herds examined. There was an average reduction in egg output of approximately 60%, and approximately 30% of 205 horses examined were shedding less than 100 eggs g-1 7 days after treatment. There was great between-herd variation of both the faecal egg-count depression (6.4-90.4%) and drug efficacy (0.0-80.0%). The non-benzimidazole drugs under investigation were fully effective against mature small strongyles.


Dynamic monitoring of membrane nanotubes formation induced by vaccinia virus on a high throughput microfluidic chip.
Membrane nanotubes (MNTs) are physical connections for intercellular communication and induced by various viruses. However, the formation of vaccinia virus (VACV)-induced MNTs has never been studied. In this report, VACV-induced MNTs formation process was monitored on a microfluidic chip equipped with a series of side chambers, which protected MNTs from fluidic shear stress. MNTs were formed between susceptible cells and be facilitated by VACV infection through three patterns. The formed MNTs varied with cell migration and virus concentration. The length of MNTs was positively correlated with the distance of cell migration. With increasing virus titer, the peak value of the ratio of MNT-carried cell appeared earlier. The immunofluorescence assay indicated that the rearrangement of actin fibers induced by VACV infection may lead to the formation of MNTs. This study presents evidence for the formation of MNTs induced by virus and helps us to understand the relationship between pathogens and MNTs.


Long-term outcome of macular microstructure assessed by optical coherence tomography in eyes with spontaneous resolution of macular hole.
To evaluate the long-term anatomic features as assessed by optical coherence tomography (OCT) in patients with spontaneously closed full-thickness macular hole (MH). Retrospective, noncomparative, observational case series. Six eyes of 6 patients who eventually showed spontaneous resolution of idiopathic MH were studied. The best-corrected visual acuity (BCVA) and OCT images of the fovea were examined before and until at least 3 years after spontaneous resolution. The MH closed spontaneously 1 to 4 months after the initial examination. There were 2 distinct macular abnormalities in the early period after spontaneous closure: 4 eyes showed outer foveal defects and the remaining 2 eyes showed persistent foveal detachment. However, the fovea regained its normal configuration and the photoreceptor inner segment/outer segment (IS/OS) junction recovered completely in all eyes. Although all 6 eyes showed visual improvement after the hole closure, the BCVA in the 4 eyes that showed delayed recovery of the IS/OS junction and foveal detachment was limited to 20/25 or worse. Spontaneous closure of MH could acquire complete anatomic reconstruction. However, delayed restoration of the IS/OS junction and persistent foveal detachment after spontaneous closure might influence the functional outcomes.


Large, organ confined, impalpable transition zone prostate cancer: association with metastatic levels of prostate specific antigen.
We present 3 of 25 patients with massive, occult transition zone cancers 7 to 86 cc in volume. Despite serum prostate specific antigen (PSA) levels of 150 to 456 ng./ml. (Yang polyclonal assay), all 3 were organ confined at radical prostatectomy and have undetectable serum PSA levels by an ultrasensitive assay at 300 to 650 days postoperatively. This clinical syndrome includes a highly elevated PSA level, benign prostatic hyperplasia on digital rectal examination, a nondiagnostic transrectal ultrasound and frequently negative transrectal or perineal needle biopsies. Clinical recognition of this syndrome plus systematic biopsies of the transition zone are the keys to diagnosis and potential cure of these patients. These cases may explain the 10% rate of men who present with metastatic prostate cancer and a normal rectal examination, much of the discrepancy between focal cancer on biopsy and large cancers in radical prostatectomy specimens, and why some men have an extraordinarily high serum PSA level and organ-confined cancer at prostatectomy.


Biases in human papillomavirus genotype prevalence assessment associated with commonly used consensus primers.
Consensus primers targeting human papillomaviruses (HPVs) have biases in sensitivity toward certain HPV types. We applied 3 primer sets (GP5+/6+, MY09/11, PGMY09/11) in parallel on 120 Chinese cervical cancer specimens. GP5+/6+ exhibited a poor sensitivity for HPV52, for which the prevalence among squamous cell cervical cancer was underestimated from 14.6% to 0%. The fact that HPV52 should rank second in prevalence among squamous cell cervical carcinoma in Hong Kong could be missed if GP5+/6+, a worldwide commonly used primer set, was selected for HPV detection. Biases in HPV type-specific sensitivity may result in misprioritization of vaccine candidates.


Honing the classification of high-risk endometrial cancer with inclusion of lymphovascular space invasion.
The purpose of this study was to analyse the clinical impact of LVSI status in a large French multicentre cohort of women with high-risk ECs according to the ESMO classification. Data of 258 women with high-risk EC who received primary surgical treatment between January 2001 and December 2012 were abstracted from prospective multicentre database. The end points were the recurrence and the lymph node metastasis (LNM) rates. Recurrence free survival (RFS) and overall survival (OS) were analyzed. The recurrence and LNM rates in the whole population were 32.9% and 34.5%, respectively. The median follow-up and initial recurrence times were 27 (range: 1-151) and 23.5 (range: 1-151) months, respectively. The respective 3-year RFS rates according to the LNM and LVSI status were, 78.3% (95% CI, 70.1-87.3), 53.7% (95% CI, 40.8-70.6), 65.5% (95% CI, 46.1-93.2), 43.5% (95% CI, 30.3-62.3) for women with no LN metastasis/no LVSI; no LN metastasis/LVSI present; LN metastasis/no LVSI; LN metastasis/LVSI present, respectively (p = 0.0005). LVSI status remains a strong prognostic factor in high-risk ECs associated with a higher recurrence rate and lower RFS and OS whatever the histological type and lymph node status. It could thus be considered in future trials to guide decision-making about adjuvant therapy in high-risk ECs.


Construction of a new polycistronic vector for over-expression and rapid purification of human hemoglobin.
To facilitate the study of the structure-function relationship of human hemoglobin (Hb A), we have developed a new hemoglobin expression vector, pGEX6P-alpha-[SD]-beta. This vector allows the co-expression of alpha-Hb as a fusion protein with Glutathione S-Transferase (GST-alpha-Hb) and beta-Hb with an additional methionine at the N-terminal extremity (rbeta-Hb). These proteins were solubilized as GST-alpha-Hb/rbeta-Hb complex form and purified in one step by affinity chromatography on immobilized glutathione. The CO binding kinetic studies show that the GST-alpha-Hb/rbeta-Hb complex and recombinant Hb A exhibit the same allosteric behavior as for native Hb A. The GST moiety, which does not modify the function of the complex, can be easily eliminated by cleavage by the PreScission Protease. After cleavage during the rapid purification procedure, over 20mg of recombinant Hb per liter of culture were obtained, more than double the yield of previous co-expression systems. This polycistronic vector system, which offers the additional advantage of a very rapid purification, is especially well suited for the study of abnormal, unstable globins in order to better understand the associated pathology.