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Abstract Single-particle cryo-EM is widely used to determine enzyme-nucleosome complex structures. However, cryo-EM sample preparation remains challenging and inconsistent due to complex denaturation at the air-water interface (AWI). Here, to address this issue, we develop graphene-oxide-coated EM grids functionalized with either single-stranded DNA (ssDNA) or thiol-poly(acrylic acid-co-styrene) (TAASTY) co-polymer. These grids protect complexes between the chromatin remodeler SNF2h and nucleosomes from the AWI and facilitate collection of high-quality micrographs of intact SNF2h-nucleosome complexes in the absence of crosslinking. The data yields maps ranging from 2.3 to 3 Å in resolution. 3D variability analysis reveals nucleotide-state linked conformational changes in SNF2h bound to a nucleosome. In addition, the analysis provides structural evidence for asymmetric coordination between two SNF2h protomers acting on the same nucleosome. We envision these grids will enable similar detailed structural analyses for other enzyme-nucleosome complexes and possibly other protein-nucleic acid complexes in general.

Abstract Burnout is a form of negative emotional and physical response to job stress. This study aimed to investigate the prevalence of burnout among healthcare workers responding to the coronavirus disease 2019 (COVID-19) outbreak in Korea and to explore correlates of burnout among healthcare workers. A nationwide questionnaire-based survey was conducted from December 1, 2020, to January 29, 2021 on 1425 healthcare workers who worked in one of the 16 healthcare facilities designated for COVID-19 care, in public health centers, or as paramedics in Korea. Burnout was assessed using 16 Korean-adapted items based on the Oldenburg Burnout Inventory (OLBI). Data were collected using a structured questionnaire and analyzed using the R version 4.1.1 software program. OLBI results indicate clinically exhaustion in 84.5% (1204/1425) and clinically disengagement in 91.1% (1298/1425), and 77.3% (1102/1425) met the score criteria for both the exhaustion and disengagement subscales for burnout. Burnout rate was significantly increased in the group with chronic fatigue symptoms (Fatigue Severity Scale ≥ 3.22) after the outbreak of COVID-19 (OR, 3.94; 95% CI 2.80–5.56), in the female group (OR, 2.05; 95% CI 1.46–2.86), in the group with physical symptoms (Patient Health Questionnaire-15 ≥ 10) after the outbreak of COVID-19 (OR, 2.03; 95% CI 1.14–3.60), in the group with a higher Global Assessment of Recent Stress scale (OR, 1.71; 95% CI 1.46–2.01), in the group with post-traumatic stress symptoms (Primary Care Post-Traumatic Stress Disorder-5 ≥ 2) (OR, 1.47; 95% CI 1.08–2.01), and in the younger age group(OR, 1.45; 95% CI 1.22–1.72). The chronic fatigue symptoms were correlated with cumulative days of care (OR, 1.18; 95% CI 1.02–1.37). The physical symptoms were correlated with average contact hours with COVID-19 patients per day (OR, 1.34; 95% CI 1.17–1.54), and cumulative days of care (OR, 1.21; 95% CI 1.06–1.38). Most Korean healthcare workers suffered from burnout related to excessive workload during the COVID-19 pandemic. During a widespread health crisis like COVID-19, it is necessary to regularly check the burnout status in healthcare workers and reduce their excessive workload by supplementing the workforce and providing appropriate working hours sufficient rest hours.

Abstract Delayed diagnosis of female genital tuberculosis (FGTB) can lead to inappropriate treatment and unnecessary surgical procedures rather than standard anti-TB medication. We tried to evaluate the use of computed tomography (CT) imaging to differentiate TB peritonitis from peritoneal carcinomatosis of advanced epithelial ovarian cancer (AEOC). We investigated women who underwent CT to distinguish between TB peritonitis and peritoneal carcinomatosis of AEOC. We evaluated various CT imaging features to identify differences between the two diseases. In addition, we performed univariate and multivariate logistic regression analyses to identify the independent imaging parameters associated with TB peritonitis and evaluated the diagnostic performance of the combined imaging parameters. We also performed the histopathological analysis of the available salpinx specimens of TB peritonitis. We included 25 women with TB peritonitis and 34 women with peritoneal carcinomatosis of AEOC. A multivariate analysis of the discriminant CT imaging features between the two diseases revealed that changes in fallopian tubes and peritoneal micronodules were independent parameters associated with TB peritonitis (p ≤ 0.012). Combining the two imaging parameters showed an area under the receiver operating characteristic curve of 0.855, a sensitivity of 88.0%, and a specificity of 67.7% for differentiating TB peritonitis from peritoneal carcinomatosis. Furthermore, changes in fallopian tubes were correlated with histopathological abnormalities in salpinx specimens. Pretreatment CT evaluation with useful imaging features could help differentiate TB peritonitis from peritoneal carcinomatosis of AEOC.

Objective We investigated the feasibility of laparoscopic restaging surgery in patients with unexpected uterine cancer. Methods This retrospective study included eight patients who underwent laparoscopic restaging surgery for Iran University uterine cancer after a prior hysterectomy or myomectomy. Results The median age of the patients and their body mass index were 55 years (range, 44–78) and 23.8 kg/m2 (range, 20.75–31.89), respectively. The median interval between the prior hysterectomy and the restaging surgery was 21 days (range, 10–35). The median operating time and time for the return of bowel activity were 325 minutes (range, 200–475) and 35 hours (range, 18–50), respectively. The median numbers of harvested pelvic and para-aortic lymph nodes were 17.5 (range, 14–29) and 20.5 (range, 7–36), respectively. In seven of the eight patients, uterine extraction was performed with vaginal or electronic morcellation. The final International Federation of Gynecology and Obstetrics stage was IA in all patients. Intraoperative and postoperative complications did not occur in any of the patients, except for the need for transfusion. Patient 4 had synchronous primary cancer (stage IA) of the endometrium and left ovary. Two of the eight patients with clear cell carcinoma received chemotherapy, and none received radiotherapy. All patients survived without disease recurrence. Conclusion Restaging surgery might be necessary for highly selective patients with unexpected uterine malignancies. This would be an alternative surgical modality for complete staging and planning tailored adjuvant treatments. However, lymphadenectomy might not be performed in patients with early uterine cancer.

Of the 141 scabies patients, 78 (55.3%) had recently received treatment, often steroids or antihistamines, but only two patients (2.6%) had been prescribed anti-scabies treatment. Sixty-eight persons had skin complaints: 21 (30.9%) and 30 (44.1%) were newly diagnosed with scabies and other skin diseases, respectively; 17 patients (25.0%) remained with persistent complaints after anti-scabies treatment of whom nine (52.9%) had incorrectly applied the treatment. Randomized trial of community treatment with azithromycin and ivermectin mass drug administration for control of scabies and impetigo. We performed multivariate mixed-effect logistic regression, adjusted for clustering within monasteries, to examine factors associated with scabies, and estimated the population attributable fraction (PAF) of impetigo that is attributable to scabies. [Extracted from the article]

IntroductionMild cognitive impairment (MCI) is often described as an intermediate stage of the normal cognitive decline associated with aging and dementia. There is a growing interest in various non-pharmacological interventions for MCI to delay the onset and inhibit the progressive deterioration of daily life functions. Previous studies suggest that cognitive training (CT) contributes to the restoration of working memory and that the brain-computer-interface technique can be applied to elicit a more effective treatment response. However, these techniques have certain limitations. Thus, in this preliminary study, we applied the neurofeedback paradigm during CT to increase the working memory function of patients with MCI.MethodsNear-infrared spectroscopy (NIRS) was used to provide neurofeedback by measuring the changes in oxygenated hemoglobin in the prefrontal cortex. Thirteen elderly MCI patients who received CT-neurofeedback sessions four times on the left dorsolateral prefrontal cortex (dlPFC) once a week were recruited as participants.ResultsCompared with pre-intervention, the activity of the targeted brain region increased when the participants first engaged in the training; after 4 weeks of training, oxygen saturation was significantly decreased in the left dlPFC. The participants demonstrated significantly improved working memory compared with pre-intervention and decreased activity significantly correlated with improved cognitive performance.ConclusionOur results suggest that the applications for evaluating brain-computer interfaces can aid in elucidation of the subjective mental workload that may create additional or decreased task workloads due to CT.

The coronavirus disease 2019 (COVID-19) outbreak became a worldwide pandemic in 2020. Social distancing measures, such as self-quarantine, lockdowns, and school closures, which have proven efficacy in various pandemic situations, remain in use in Korea. These measures prevented viral transmission to some extent; however, adverse effects have also resulted. First, the negative effect of social isolation on mental health is evident. This influences the psychiatric milieu of parents and children directly and indirectly. The most stressful factor among Korean youth was the restriction of outdoor activities. Increasing parenting burden result in increased screen time among youth, and social isolation created depressive mood with symptoms similar to those of attention deficit hyperactivity disorder and anxiety. Second, symptoms of posttraumatic stress disorder (PTSD) and somatization are prevalent among children and adolescents. The sense of threatened health and life during the pandemic, one symptom of PTSD, is a strong risk factor for somatization. Finally, the increased pattern of child abuse in pandemic indicates increased levels of emotional/psychological abuse and nonmedical neglect. Social isolation makes people less aware of these events. Because pediatricians evaluate pediatric patients and their families, they should regularly assess emotional/stress factors, especially when somatization is prominent during the pandemic, and cautiously recommend that families seek advice from mental health professionals when warranted. Primary physicians must understand the characteristics and aspects of child abuse in the COVID-19 pandemic, make efforts to identify signs of child abuse, and deliver accurate information and preventive strategies for child abuse to caregivers, thereby functioning as a professional guardian. To promote the mental health of parents and children during the COVID-19 pandemic, more research and cooperation among health professionals, families, governments, and schools are needed in the future.

Abstract The SARS-CoV-2 pandemic is rapidly evolving and remains a major health challenge worldwide. With an increase in pregnant women with COVID-19 infection, we recognized an urgent need to set up a multidisciplinary taskforce to provide safe and holistic care for this group of women. In this review of practice in a tertiary hospital in Singapore, we discuss the key considerations in setting up an isolation maternity unit and our strategies for peripartum and postpartum care. Through teleconsultation, we involve these women and their families in the discussion of timing and mode of birth, disposition of babies after birth and safety of breastfeeding to enable them to make informed decisions and individualize their care.

Abstract Background Multiple gestations are associated with an increased incidence of preeclampsia. However, there exists no evidence for an association between multiple gestations and development of hypertension(HTN) later in life. This study aimed to determine whether multiple gestations are associated with HTN beyond the peripartum period. Methods In this retrospective nationwide population-based study, women who delivered a baby between January 1, 2007, and December 31, 2008, and underwent a national health screening examination within one year prior to their pregnancy were included. Subsequently, we tracked the occurrence of HTN during follow-up until December 31, 2015, using International Classification of Diseases-10th Revision codes. Results Among 362,821 women who gave birth during the study period, 4,944 (1.36%) women had multiple gestations. The cumulative incidence of HTN was higher in multiple gestations group compared with singleton group (5.95% vs. 3.78%, p

Introduction: During resuscitations, healthcare professionals (HCPs) find balancing the need for timely resuscitation and adherence to infection prevention (IP) measures difficult. This study explored the effects of an innovative teaching method, using in-situ simulation and inter-professional education to enhance compliance to IP through better inter-professional collaboration. Methods: The study was conducted in the Surgical Intensive Care Unit (SICU) in a 1200-beds teaching hospital. HCPs working in the SICU were conveniently allocated to the intervention or control group based on their work roster. The intervention group attended an in-situ simulated scenario on managing cardiac arrest in an infectious patient. The control group completed the standard institution-wide infection control eLearning module. Outcomes measured were: (a) attitudes towards inter-professional teamwork [TeamSTEPPS Teamwork Attitudes Questionnaire (TAQ)], (b) infection prevention knowledge test, (c) self-evaluated confidence in dealing with infectious patients and (d) intensive care unit (ICU) audits on infection prevention compliance during actual resuscitations. Results: 40 HCPs were recruited. 29 responded (71%) to the pre- and post-workshop questionnaires. There were no significant differences in the TeamSTEPPS TAQ and infection prevention knowledge score between the groups. However, ICU audits demonstrated a 60% improvement in IP compliance for endotracheal tube insertion and 50% improvement in parenteral medication administration. This may be attributed to the debriefing session where IP staff shared useful tips on compliance to IP measures during resuscitation and identified threats that could deter IP compliance in SICU. Conclusion: Learning infection prevention through simulated inter-professional education (IPE) workshops may lead to increased IP compliance in clinical settings.

Abstract An electron-rich fused indoloindole-based poly(indoloindole-selenophene vinylene selenophene) was synthesized and characterized. Soxhlet can be obtained by continuously purifying the product with a specific solvent and obtaining a pure polymer with a high concentration. Molecular weight is affected by the vapor pressure of marginal solvent, and the polymer was fractionated using tetrahydrofuran, chloroform, and chlorobenzene. Solubility is closely related to the morphology of bulk heterojunction and device parameters. In the solution process of fabricating the organic solar cell, securement of solubility has a great effect on the performance of the device, because morphology and orientation of a photo-active layer which significantly affect charge transport in the device. Since tetrahydrofuran (THF) Soxhlet solvents have high vapor pressure and appropriate solubility parameters, THF induced the best solubility of P-IDI-SVS materials for organic solvents. And through additive optimization, the performance of the device based on P-IDI-SVS from THF-Soxhlet extraction was enhanced. This is expected to be a meaningful study because the effect on solubility of Soxhlet solvent suggests factors to be considered in the solution process in organic solar cell research. In addition, surface modified bulk heterojunction was observed using atomic force microscopy, photoluminescence, time-correlated single photon counting and Raman spectroscopy analysis.

Evidence has emerged that pregnant women who contract coronavirus disease 2019 (Covid-19) are at increased risk of certain forms of severe illness as well as complications requiring intensive care unit admission and resultant mortality. Teleconsultations can facilitate continuing care for obstetric patients during the Covid-19 pandemic while reducing their risk of exposure to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In this short report, we share our experience in the provision of teleconsultations for ambulatory obstetric anaesthesia patients in our high-risk obstetric anaesthesia clinic during the Covid-19 pandemic. Appropriate labour analgesia or anaesthesia plans were able to be formulated and communicated to the patients by teleconsultation, resulting in no delay or compromise in their peripartum care. Both patients and clinicians reported satisfaction with the teleconsultation process and outcome. The considerations and challenges in setting up a teleconsultation service as well as the factors in favour of teleconsultation are also explored.

Pulsed EPR dipolar spectroscopy is a powerful tool for determining the structure and conformational dynamics of biological macromolecules, as it allows precise measurements of distances in the range of 1.5–10 nm. Utilization of high-spin Mn2+ species as spin probes for distance measurements is of significant interest, because they are biologically compatible and endogenous in numerous biological systems. However, to date dipolar spectroscopy experiments with this kind of species have been underexplored. Here we present pulsed electron electron double resonance (PELDOR also called DEER) and relaxation-induced dipolar modulation enhancement (RIDME) experiments, which have been performed at W-band (94 GHz) and J-band frequencies (263 GHz) on a bis-MnDOTA (DOTA = 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate) model system. The distances obtained from these experiments are in good agreement with predictions. RIDME experiments reveal a significantly higher modulation depth compared to PELDOR, which is an important consideration for biological samples. These experiments also feature higher harmonics of the dipolar coupling frequency due to effective multiple-quantum relaxation of high-spin Mn2+ as well as the multiple-component background function. Harmonics of the dipolar coupling frequency were taken into account by including additional terms in the kernel function of Tikhonov regularization analysis. [ABSTRACT FROM AUTHOR]

ObjectiveThis study aimed to determine the association between preeclampsia and the postpartum development of metabolic syndrome based on the pre-pregnancy status.MethodsKorean women who delivered their first child between January 1, 2011, and December 31, 2012, were enrolled. All subjects underwent a national health screening examination conducted by the National Health Insurance Corporation 1 or 2 years prior to their first delivery and within 2 years after their first delivery.ResultsAmong the 49,065 participants, preeclampsia developed in 3,391 participants (6.9%). The prevalence of metabolic syndrome was higher postpartum in women with preeclampsia than in those without preeclampsia (4.9% vs. 2.7%, respectively, P

Matrix metalloproteinase-11 (MMP-11) promote cancer invasion and metastasis through degrading the extracellular matrix. Protein degradation by MMP-11 in tumor cells may progressively suppress cancer surveillance activities with blocking immune response in breast cancer. The aim of study is to analyze clinicopathological parameters, molecular interactions and anticancer immune response in patients with MMP-11 expression and to provide candidate target drugs. We investigated the clinicopathologic parameters, specific gene sets, tumor antigenicity, and immunologic relevance according to MMP-11 expression in 226 and 776 breast cancer patients from the Hanyang University Guri Hospital (HUGH) cohort and The Cancer Genome Atlas (TCGA) data, respectively. We analyzed pathway networks and in vitro drug response. High MMP-11 expression was associated with worse survival rate in breast cancer from HUGH cohort and TCGA data (all p < 0.05). In analysis of immunologic gene sets, high MMP-11 expression was related to low immune response such as CD8+T cell, CD4+T cell and B cell. In silico cytometry, there was a decrease of cancer testis antigen and low tumor infiltrating lymphocyte in patient with high MMP-11 expression: activated dendritic cell, CD8+T cell, CD4+ memory T cell, and memory B cell. In pathway networks, MMP-11 was linked to the pathways including low immune response, response to growth hormone and catabolic process. We found that pictilisib and AZ960 effectively inhibited the breast cancer cell lines with high MMP-11 expression. Strategies making use of MMP-11-related hub genes could contribute to better clinical management/research for patients with breast cancer.

The purpose of the paper is to examine the meaning of 'lifelong learning to be' as the essence of lifelong education, which has been implied in the thoughts of Friedrich Wilhelm Nietzsche (1844-1900). This will be approached from the perspectives of 'learning to know', 'learning to do', 'learning to live together' and 'learning to be', which are the four pillars of education in UNESCO's 1996 Delors Report. Despite Friedrich Nietzsche being one of the most influential scholars of the nineteenth century, few types of research have been carried out concerning his philosophy and its impact on lifelong education. This article, first and foremost, seeks to argue that Nietzsche is a significantly prominent lifelong educator whose philosophy contains significant implications on lifelong learning in today's period of uncertainty. Secondly, we will discuss the 'learning to be' as the pillar of lifelong learning suggested in both Faure and Delors Report. In the final part, the article explores the implications of Nietzsche's idea on the four pillars of lifelong learning.

Accurate protein localization is crucial to generate and maintain organization in all cells. Achieving accuracy is challenging, as the molecular signals that dictate a protein’s cellular destination are often promiscuous. A salient example is the targeting of an essential class of tail-anchored (TA) proteins, whose sole defining feature is a transmembrane domain near their C-terminus. Here we show that the Guided Entry of Tail-anchored protein (GET) pathway selects TA proteins destined to the endoplasmic reticulum (ER) utilizing distinct molecular steps, including differential binding by the co-chaperone Sgt2 and kinetic proofreading after ATP hydrolysis by the targeting factor Get3. Further, the different steps select for distinct physicochemical features of the TA substrate. The use of multiple selection filters may be general to protein biogenesis pathways that must distinguish correct and incorrect substrates based on minor differences.

• High prevalence of transmitted drug resistance (TDR) in acute HIV-1 patients • Good efficacy of dolutegravir (DTG)-based regimen even in the presence of TDR • Time to viro-suppression was similar for DTG and non-DTG-based regimens • Interruptions of DTG-based regimen were rare The aim of this study was to compare the tolerability and viro-immunological efficacy of dolutegravir-based regimens (DTG group) with regimens based on EVG, RAL, PI or NNRTI (NODTG group) in patients with acute HIV-1 infections (AHI). All patients diagnosed with AHI and on antiretroviral therapy (ART) between January 2015 and December 2017 from five centers in Italy were included and followed-up to 30th April 2018. AHI was defined by the presence of the positive p24 antigen with negative or indeterminate western blot. Forty-three patients were enrolled: 20 in the DTG group, 23 in the NODTG group. Nine patients (20.9%; four in the DTG group, five in the NODTG group) were prescribed a four-drug regimen. In the cohort, 81.4% were Italian and 83.7% were male, with a median age of 41 years (interquartile range [IQR] 31-48). Median time between HIV diagnosis and ART initiation was 12 days (IQR 5-28). Seven patients harbored a virus with transmitted mutations at baseline (16.2%), all were in the DTG group (P =0.005). All patients had undetectable HIV-RNA at the end of follow-up except two patients, one of whom had 57 copies and one who was lost to follow-up. In Kaplan-Meier analysis, time to virological suppression was similar in the two groups (log rank: P = 0.7155). After achieving virological suppression, four patients stopped ART because of toxicity: two on DTG, two on EVG for neurological and gastrointestinal toxicity, respectively. In our setting, ART in AHI is started very early. DTG showed good viro-immunological efficacy even in the presence of NRTI-transmitted mutations. DTG interruptions were rare. [ABSTRACT FROM AUTHOR]

Our aim was to determine the impact of antimicrobial stewardship tools (ASTs) and the COVID-19 pandemic on antibiotic consumption (AC). We used the national software Consores ® to determine AC in DDD/1000 days of hospitalization from 2017 to 2022 in voluntary private hospitals in France. The ASTs considered were: 1. internal guidelines; 2. the list of antibiotics with restricted access; 3. the presence of an antibiotic referent or 4. an ID specialist; and 5. proof of an annual meeting on antimicrobial resistance. Institutions with dedicated units for COVID-19 patients were specified. In 30 institutions, the total AC varied from (means) 390 to 405 DDD/1000 DH from 2017 to 2022. Fluoroquinolones and amoxicillin/clavulanate consumption decreased from 50 to 36 ( p = 0.003) and from 112 to 77 ( p = 0.025), respectively, but consumption of piperacillin/tazobactam increased from 9 to 21 ( p < 0.001). Over the study period, 10 institutions with ≤2 AST had lower AC compared to 20 institutions with ≥3 AST ( p < 0.01). COVID-19 units opened in 10 institutions were associated with a trend toward higher macrolide consumption from 15 to 25 from 2017 to 2020 ( p = 0.065) and with an acceleration of piperacillin/tazobactam consumption from 2020 to 2022 ( p ≤ 0.003). Antibiotic consumption in 30 private hospitals in France was inversely related to the number of AST. The COVID-19 pandemic was associated with limited impact on AC, but special attention should be paid to piperacillin/tazobactam consumption.

Genetic code expansion allows modification of the physical and chemical properties of proteins by the site-directed insertion of noncanonical amino acids. Here we exploit this technology for measuring nanometer-scale distances in proteins. (2,2'-Bipyridin-5-yl)alanine was incorporated into the green fluorescent protein (GFP) and used as an anchoring point for Cu(II) to create a spin-label. The incorporation of (2,2'-bipyridin-5-yl)alanine directly into the protein resulted in a high-affinity binding site for Cu(II) capable of outcompeting other binding positions in the protein. The resulting Cu(II)-spin label is very compact and not larger than a conventional amino acid. By using 94 GHz electron paramagnetic resonance (EPR) pulse dipolar spectroscopy we have been able to determine accurately the distance between two such spin-labels. Our measurements revealed that GFP dimers can adopt different quaternary conformations. The combination of spin-labeling using a paramagnetic nonconventional amino acid with high-frequency EPR techniques resulted in a sensitive method for studying the structures of proteins.

Quantum information technology puts stringent demands on the quality of materials and interfaces in the pursuit of increased device coherence. Yet, little is known about the chemical structure and origins of paramagnetic impurities that produce flux/charge noise that causes decoherence of fragile quantum states and impedes the progress toward large-scale quantum computing. Here, we perform high magnetic field electron paramagnetic resonance (HFEPR) and hyperfine multispin spectroscopy on α-Al 2 O 3 , a common substrate for quantum devices. In its amorphous form, α-Al 2 O 3 is also unavoidably present in aluminum-based superconducting circuits and qubits. The detected paramagnetic centers are immanent to the surface and have a well-defined but highly complex structure that extends over multiple hydrogen, aluminum, and oxygen atoms. Modeling reveals that the radicals likely originate from well-known reactive oxygen chemistry common to many metal oxides. We discuss how EPR spectroscopy might benefit the search for surface passivation and decoherence mitigation strategies.

This is the first study to examine the effects of in vitro digestion on biological activities of Sargassum spp., a broadly known brown seaweed for therapeutic potential. Three fractions (F1-F3) were obtained from hexane extract by column chromatography. Under in vitro simulated digestion, the anti-α-amylase capacity of F1 in oral and intestinal phases increases, while it significantly decreases in the gastric phase. The α-amylase inhibition of F2 promotes throughout all digestive stages while the activity of F3 significantly reduces. The cytotoxic activity of F1 against U266 cell-line accelerates over the oral, gastric, and intestinal stages. The fractions F2 and F3 exhibited the declined cytotoxic potentialities in oral and gastric phases, but they were strengthened under intestinal condition. Palmitic acid and fucosterol may play an active role in antidiabetic and cytotoxic activity against multiple myeloma U266 cell line of Sargassum spp. However, the involvement of other phytochemicals in the seaweed should be further investigated.

We have generated a mutant of C. elegans manganese superoxide dismutase at histidine 30 by site-directed mutagenesis. The structure was solved at a resolution of 1.52 Å by X-ray crystallography (pdb: 6S0D). His30 was targeted, as it forms as a gateway residue at the top of the solvent access funnel to the active site, together with Tyr34. In the wild-type protein, these gateway residues are involved in the hydrogen-bonding network providing the protons necessary for the catalytic reaction at the metal center. However, biophysical characterization and cell viability experiments reveal that a mutation from histidine to asparagine in the H30N mutant modifies metal selectivity in the protein, favoring the uptake of iron over manganese in minimal media conditions, alters active-site coordination from the characteristic trigonal bipyramidal to octahedral geometry, and encourages cellular proliferation in K562 cells, when added exogenously to the cells.

The 285 GHz EPR spectra of perchlorotriphenylmethyl and tetrathiatriarylmethyl radicals in frozen solution have been accurately measured. The relationship between their molecular structures and their g-tensors has been investigated with the aid of DFT calculations, revealing that the degree of spin density delocalization away from the central methylene carbon is an important determining factor of the g-anisotropy. In particular, the small amount of spin densities on the Cl or S heteroatoms at the 2 and 6 positions with respect to the central carbon have the strongest influence. Furthermore, the amount of spin densities on these heteroatoms and thus the anisotropy can be modulated by the protonation (esterification) state of the carboxylate groups at the 4 position. These results provide unique insights into the g-anisotropy of persistent trityl radicals and how it can be tuned.

Almost half of all enzymes utilize a metal cofactor. However, the features that dictate the metal utilized by metalloenzymes are poorly understood, limiting our ability to manipulate these enzymes for industrial and health-associated applications. The ubiquitous iron/manganese superoxide dismutase (SOD) family exemplifies this deficit, as the specific metal used by any family member cannot be predicted. Biochemical, structural and paramagnetic analysis of two evolutionarily related SODs with different metal specificity produced by the pathogenic bacterium Staphylococcus aureus identifies two positions that control metal specificity. These residues make no direct contacts with the metal-coordinating ligands but control the metal's redox properties, demonstrating that subtle architectural changes can dramatically alter metal utilization. Introducing these mutations into S. aureus alters the ability of the bacterium to resist superoxide stress when metal starved by the host, revealing that small changes in metal-dependent activity can drive the evolution of metalloenzymes with new cofactor specificity.

To elucidate biochemical mechanisms leading to seed deterioration, we studied 23 wheat genotypes after exposure to seed bank storage for 6-16 years compared to controlled deterioration (CD) at 45 °C and 14 (CD14) and 18% (CD18) moisture content (MC) for up to 32 days. Under two seed bank storage conditions, seed viability was maintained in cold storage (CS) at 0 °C and 9% seed MC, but significantly decreased in ambient storage (AS) at 20 °C and 9% MC. Under AS and CS, organic free radicals, most likely semiquinones, accumulated, detected by electron paramagnetic resonance, while the antioxidant glutathione (GSH) was partly lost and partly converted to glutathione disulphide (GSSG), detected by HPLC. Under AS the glutathione half-cell reduction potential (E GSSG/2GSH ) shifted towards more oxidising conditions, from -186 to -141 mV. In seeds exposed to CD14 or CD18, no accumulation of organic free radicals was observed, GSH and seed viability declined within 32 and 7 days, respectively, GSSG hardly changed (CD14) or decreased (CD18) and E GSSG/2GSH shifted to -116 mV. The pH of extracts prepared from seeds subjected to CS, AS and CD14 decreased with viability, and remained high under CD18. Across all treatments, E GSSG/2GSH correlated significantly with seed viability ( r  = 0.8, p <.001). Data are discussed with a view that the cytoplasm is in a glassy state in CS and AS, but during the CD treatments, underwent transition to a liquid state. We suggest that enzymes can be active during CD but not under the seed bank conditions tested. However, upon CD, enzyme-based repair processes were apparently outweighed by deteriorative reactions. We conclude that seed ageing by CD and under seed bank conditions are accompanied by different biochemical reactions.

VUV photons from a synchrotron source were used to record the gas-phase absorption spectrum of isoquinoline over the range 3.5 to 10.7 eV. The rich spectrum exhibits both broad and sharp features, of varying intensities, that are analyzed into eight valence and eight Rydberg transitions. Previous data on the valence transitions of isoquinoline were essentially limited to solution spectra up to 5.4 eV. Our study increases their number considerably. The features in the 3.96 eV region are discussed in terms of vibronic coupling between the nπ* 1 1 A'' and ππ* 2 1 A' valence electronic states. The intensities of some spectral features are augmented by collective π-electron modes considered to be of plasmon-type. Assignments of the valence transitions were facilitated by our DFT calculations and by earlier Pariser-Parr-Pople MO calculations. The calculation results are compared and their relative value is discussed. The DFT calculations reproduce very well a number of experimentally determined properties of the ground state of isoquinoline, in particular its bond distances and angles, rotational constants, vibrational frequencies and dipole moment. No Rydberg series of isoquinoline have previously been observed. Three of the newly observed Rydberg series converge to the D 0 electronic ground state of the ion, while two converge to the D 1 and three to the D 3 excited electronic states of the cation. Astrophysical applications of the VUV absorption spectrum of isoquinoline, in particular the measured absorption cross-sections, are briefly discussed. A comparison between the absorption spectra of isoquinoline and quinoline highlights their similarities and differences, related to their respective molecular orbitals., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

Mn 2+ often serves as a paramagnetic substitute to Mg 2+ , providing means for exploring the close environment of Mg 2+ in many biological systems where it serves as an essential co-factor. This applies to proteins with ATPase activity, where the ATP hydrolysis requires the binding of Mg 2+ -ATP to the ATPase active site. In this context, it is important to distinguish between the Mn 2+ coordination mode with free ATP in solution as compared to the protein bound case. In this work, we explore the Mn 2+ complexes with ATP, the non-hydrolysable ATP analog, AMPPNP, and ADP free in solution. Using W-band 31 P electron-nuclear double resonance (ENDOR) we obtained information about the coordination to the phosphates, whereas from electron-electron double resonance (ELDOR) - detected NMR (EDNMR) we determined the coordination to an adenosine nitrogen. The coordination to these ligands has been reported earlier, but whether the nitrogen and phosphate coordination is within the same nucleotide molecules or different ones is still under debate. By applying the correlation technique, THYCOS (triple hyperfine correlation spectroscopy), and measuring 15 N- 31 P correlations we establish that in Mn-ATP in solution both phosphates and a nitrogen are coordinated to the Mn 2+ ion. We also carried out DFT calculations to substantiate this finding. In addition, we expanded the understanding of the THYCOS experiment by comparing it to 2D-EDNMR for 55 Mn- 31 P correlation experiments and through simulations of THYCOS and 2D-EDNMR spectra with 15 N- 31 P correlations., (Copyright © 2018 Elsevier Inc. All rights reserved.)

The absorption spectrum of quinoline was measured in the gas phase between 3.5 and 10.7 eV using a synchrotron photon source. A large number of sharp and broad spectral features were observed, some of which have plasmon-type collective π-electron modes contributing to their intensities. Eight valence electronic transitions were assigned, considerably extending the number of π-π* transitions previously observed mainly in solution. The principal factor in solution red-shifts is found to be the Lorentz-Lorenz polarizability parameter. Rydberg bands, observed for the first time, are analyzed into eight different series, converging to the D 0 ground and two excited electronic states, namely, D 3 and D 4 , of the quinoline cation. The R1 series limit is 8.628 eV for the first ionization energy of quinoline, a value more precise than previously published. This value, combined with cation electronic transition data, provides precise energies, respectively, 10.623 and 11.355 eV, for the D 3 and D 4 states. The valence transition assignments are based on density functional theory (DFT) calculations as well as on earlier Pariser-Parr-Pople (P-P-P) self-consistent field linear combination of atomic orbitals molecular orbital results. The relative quality of the P-P-P and DFT data is discussed. Both are far from spectroscopic accuracy concerning electronic excited states but were nevertheless useful for our assignments. Our time-dependent DFT calculations of quinoline are excellent for its ground-state properties such as geometry, rotational constants, dipole moment, and vibrational frequencies, which agree well with experimental observations. Vibrational components of the valence and Rydberg transitions mainly involve C-H bend and C═C and C═N stretch modes. Astrophysical applications of the vacuum UV absorption of quinoline are briefly discussed.

The pathogenicity of Staphylococcus aureus is enhanced by having two superoxide dismutases (SODs): a Mn-specific SOD and another that can use either Mn or Fe. Using 94 GHz electron-nuclear double resonance (ENDOR) and electron double resonance detected (ELDOR)-NMR we show that, despite their different metal-specificities, their structural and electronic similarities extend down to their active-site 1 H- and 14 N-Mn(ii) hyperfine interactions. However these interactions, and hence the positions of these nuclei, are different in the inactive Mn-reconstituted Escherichia coli Fe-specific SOD. Density functional theory modelling attributes this to a different angular position of the E. coli H171 ligand. This likely disrupts the Mn-H171-E170' triad causing a shift in charge and in metal redox potential, leading to the loss of activity. This is supported by the correlated differences in the Mn(ii) zero-field interactions of the three SOD types and suggests that the triad is important for determining metal specific activity.

Background: Prostate cancer is the most common solid tumor. The incidence of prostate cancer shows regional and racial differences. The ideal PSA threshold for prostate biopsy is still being debated., Objective: We aimed to investigate cancer detection rates in Turkish men who underwent transrectal ultrasound-guided prostate biopsy (TRUSPB) who had prostate-specific antigen (PSA) levels in the range of 2.5 to 4.0 ng/mL and compare them with the rates of cancer in patients with PSA levels in the range of 4.0 to 10.0 ng/mL., Methods: All Turkish men who underwent TRUSPB in our clinic between January 2012 and May 2014 were included; that is, 101 patients (Group 1) with PSA level in the range of 2.5 to 4.0 ng/mL and 522 patients (Group 2) with PSA level in the range of 4.0 to 10.0 ng/mL. Mean PSA level, age, prostate volume, and cancer detection rates were evaluated., Results: The mean age was 60.5 and 64 years in Group 1 and Group 2, respectively ( P = 0.06). The mean PSA level was determined as 3.1 and 6.8 ng/mL in Group 1 and Group 2, respectively ( P = 0.03). The cancer detection rate was 12.7% in Group 1 (n = 13) and 30.8% in Group 2 (n = 161), which revealed a statistically significant difference between the 2 groups ( P = 0.001). In Group 1, 9 of 13 patients (69%) had Gleason score of 6, 3 (23%) had Gleason score of 7, and 1 (8%) had a Gleason score of 8., Conclusions: The cancer detection rate is lower in Turkish men with PSA level in the range of 2.5 to 4.0 ng/mL when compared with men with PSA level in the range of 4.0 to 10.0 ng/mL. Furthermore, most patients in whom cancer was detected who have a PSA level in the range of 2.5 to 4.0 ng/mL are low risk. Therefore, the benefit of TRUSBP in Turkish men with PSA level between 2.5 and 4 ng/mL is low.

Pulsed EPR dipolar spectroscopy is a powerful tool for determining the structure and conformational dynamics of biological macromolecules, as it allows precise measurements of distances in the range of 1.5-10 nm. Utilization of high-spin Mn 2+ species as spin probes for distance measurements is of significant interest, because they are biologically compatible and endogenous in numerous biological systems. However, to date dipolar spectroscopy experiments with this kind of species have been underexplored. Here we present pulsed electron electron double resonance (PELDOR also called DEER) and relaxation-induced dipolar modulation enhancement (RIDME) experiments, which have been performed at W-band (94 GHz) and J-band frequencies (263 GHz) on a bis-MnDOTA (DOTA = 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate) model system. The distances obtained from these experiments are in good agreement with predictions. RIDME experiments reveal a significantly higher modulation depth compared to PELDOR, which is an important consideration for biological samples. These experiments also feature higher harmonics of the dipolar coupling frequency due to effective multiple-quantum relaxation of high-spin Mn 2+ as well as the multiple-component background function. Harmonics of the dipolar coupling frequency were taken into account by including additional terms in the kernel function of Tikhonov regularization analysis.

Double electron-electron resonance (DEER) can be used to study the structure of a protein in its native cellular environment. Until now, this has required isolation, in vitro labeling, and reintroduction of the protein back into the cells. We describe a completely biosynthetic approach that avoids these steps. It exploits genetically encodable lanthanide-binding tags (LBT) to form self-assembling Gd(III) metal-based spin labels and enables direct in-cell measurements. This approach is demonstrated using a pair of LBTs encoded one at each end of a 3-helix bundle expressed in E. coli grown on Gd(III) -supplemented medium. DEER measurements directly on these cells produced readily detectable time traces from which the distance between the Gd(III) labels could be determined. This work is the first to use biosynthetically produced self-assembling metal-containing spin labels for non-disruptive in-cell structural measurements., (© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Recent studies have shown that in spite of being generally close to the equator; vitamin D deficiency is common in South East Asian countries. In order to quantify micronutrient status for women and children in Cambodia; a nationally-representative survey was conducted in 2014 linked to the Cambodian Demographic Health Survey. The countrywide median of 25(OH)D was, respectively, 64.9 and 91.1 nmol/L for mothers and children. Based on The Endocrine Society cutoffs (>50<75 nmol/L = insufficiency; ≤50 nmol/L = deficiency); 64.6% of mothers and 34.8% of their children had plasma vitamin D concentrations indicating insufficiency or deficiency. For deficiency alone, 29% of the mothers were found to be vitamin D deficient, but only 13.4% of children. Children who live in urban areas had a 43% higher rate of vitamin D insufficiency versus those who live in rural areas (OR; 1.434; 95% CI: 1.007; 2.041). However, such differences were not observed in their mothers. The high prevalence of vitamin D deficiency is likely in part due to lifestyle choices, including sun avoidance, increasingly predominant indoor work, and covered transport. These survey findings support the need for a broader national Cambodian study incorporating testing of adult men, adolescents and the elderly, and encompassing other parameters such as skeletal health. However, the data presented in this study already show significant deficiencies which need to be addressed and we discuss the benefit of establishing nationally-mandated food fortification programs to enhance the intake of vitamin D.

A 2014 national assessment of salt iodization coverage in Cambodia found that 62% of samples were non-iodized, suggesting a significant decline in daily iodine intakes. The Cambodian Micronutrient Survey conducted in 2014 (CMNS-2014) permitted obtaining national data on urinary iodine concentrations (UIC) to assess iodine status and whether iodized salt use had an impact. Urine samples were collected from mothers (n = 736) and children (n = 950). The median UIC was 63 µg/L and 72 µg/L in mothers and children respectively. More than 60% of mothers and their children had a UIC < 100 µg/L, thereby indicating a serious public health problem. Iodine status was significantly lower among mothers and children living in rural areas, belonging to the poorest socioeconomic category, or living in a household not using iodized salt. The limited enforcement of the legislation for iodized salt has resulted in a major decrease in the prevalence of iodized salt, which in turn has compromised iodine status in Cambodia. It is essential for the government to enhance enforcement of the iodized salt legislation, and implement short term strategies, such as iodine supplementation, to prevent an increase of severe complications due to iodine deficiency in the Cambodian population.

Manganous phosphates have been postulated to play an important role in cells as antioxidants. In situ Mn(II) electron-nuclear double resonance (ENDOR) spectroscopy has been used to measure their speciation in cells. The analyses of such ENDOR spectra and the quantification of cellular Mn(II) phosphates has been based on comparisons to in vitro model complexes and heuristic modeling. In order to put such analyses on a more physical and theoretical footing, the Mn(II)-(31)P hyperfine interactions of various Mn(II) phosphate complexes have been measured by 95 GHz ENDOR spectroscopy. The dipolar components of these interactions remained relatively constant as a function of pH, esterification, and phosphate chain length, while the isotropic contributions were significantly affected. Counterintuitively, although the manganese-phosphate bonds are weakened by protonation and esterification, they lead to larger isotropic values, indicating higher unpaired-electron spin densities at the phosphorus nuclei. By comparison, extending the phosphate chain with additional phosphate groups lowers the spin density. Density functional theory calculations of model complexes quantitatively reproduced the measured hyperfine couplings and provided detailed insights into how bonding in Mn(II) phosphate complexes modulates the electron-spin polarization and consequently their isotropic hyperfine couplings. These results show that various classes of phosphates can be identified by their ENDOR spectra and provide a theoretical framework for understanding the in situ (31)P ENDOR spectra of cellular Mn(II) complexes.

The ability to characterize the structure of metal centers beyond their primary ligands is important to understanding their chemistry. High-magnetic-field pulsed electron double resonance detected NMR (ELDOR-NMR) is shown to be a very sensitive approach to measuring the multinuclear NMR spectra of the nuclei surrounding Mn(II) ions. Resolved spectra of intact organisms with resonances arising from (55)Mn, (31)P, (1)H, (39)K, (35)Cl, (23)Na, and (14)N nuclei surrounding Mn(2+) centers were obtained. Naturally abundant cellular (13)C could be routinely measured as well. The amplitudes of the (14)N and (2)H ELDOR-NMR spectra were found to be linearly dependent on the number of nuclei in the ligand sphere. The evolution of the Mn(II) ELDOR-NMR spectra as a function of excitation time was found to be best described by a saturation phenomenon rather than a coherently driven process. Mn(II) ELDOR-NMR revealed details about not only the immediate ligands to the Mn(II) ions but also more distant nuclei, providing a view of their extended structures. This will be important for understanding the speciation and chemistry of the manganese complexes as well as other metals found in organisms.

Pulse electron-electron double resonance (PELDOR) is a versatile technique for probing the structures and functions of complex biological systems. Despite the recent interest in high-spin metal-ions for high field/frequency applications, PELDOR measurements of Mn(ii) remain relatively underexplored. Here we present Mn(ii)-Mn(ii) PELDOR distance measurements at 94 GHz on polyproline II (PPII) helices doubly spin-labeled with Mn(ii)DOTA, which are distinguished by their small zero-field interaction. The measured Mn-Mn distances and distribution profiles were in good agreement with the expected values from molecular models. Additional features in the frequency-domain spectra became apparent at certain combinations of detect and pump frequencies. Spin-Hamiltonian calculations showed that they likely arose from contributions from the pseudo-secular component of the dipolar interaction that were found to be non-negligible for Mn(ii)DOTA. However, the influence of the pseudo-secular component on the distance distribution profiles apparently was limited. The results show the potential of Mn(ii)DOTA spin labels for high-field PELDOR distance measurements in proteins and other biological systems.

Introduction: Percutaneous nephrolithotomy (PCNL) is the primary surgical intervention in kidney stone management. Even though it is performed quite often, the complication rates are also high. Arteriovenous fistulas following extended hemorrhages after PCNL are one of the most serious complications of this operation. Our main objective was to review the data of patients who required angiography and embolization., Methods: In total, we included 1405 patients who underwent PCNL between 2007 and 2014. All patient data were retrospectively reviewed. All patients went under PCNL using fluoroscopy. Following informed consent, all hemorrhagic patients underwent angiography in the interventional radiology department and embolization was performed in patients with a hemorrhage focus point., Results: A total of 147 patients (10.4%) required transfusion for post-PCNL hemorrhages. Of them, 14 (0.99%) underwent angiography and embolization (9 [64.2%] were male and 5 [35.8%] were female, with a mean age of 39.4 ± 10.2). The remaining 133 patients were conservatively managed (81 [60.9%] males and 52 [39.1%] females, with a mean age of 42.3 ± 12.4). When the predicting factors for angiography and embolization were reviewed, renal abnormalities and the mean size of stones were significant in both univariate and multivariate analysis (p < 0.001)., Conclusion: Patients with extended and intermittent hematuria should be monitored closely for hemodynamics; if there is an ongoing necessity for transfusion, angiography should be considered.

Introduction: Radical prostatectomy (RP) is considered the best treatment for the management of localized prostate cancer in patients with life expectancy over 10 years. However, a complete recovery is not guaranteed for all patients who received/underwent RP treatment. Biochemical recurrence is frequently observed during the post-operative follow-up period. The main objective in this study is to evaluate the predictive factors of biochemical recurrence in localized prostate cancer patients who underwent RP surgery., Material and Methods: The study included 352 patients with prostate cancer treated by RP at a single institution between February 2004 and June 2014. Detailed pathological and follow-up data of all patients were obtained and analyzed to determine the results., Results: Mean follow-up duration was 39.7 months. 83 patients (23%) experienced biochemical recurrence (BCR) during the follow-up period. Mean BCR duration range was 6.56 (1-41) months. In multivariate logistic regression analysis, Gleason score (GS), PSA and extra-capsular tumour spread (ECS) variables were found to be statistically significant as BCR predictive factors., Conclusions: According to our study results, it is thought that PSA, GS and ECS can all be used for guidance in choosing a treatment modality for post-RP biochemical recurrence and metastatic disease as predictive factors. However, there is no consensus in this matter and it is still debated.

Castleman's disease (CD) is a non-clonal lymph node hyperplasia, mostly seen in the mediastinum. It has various clinical and pathological outcomes. There are different treatments because of its rare occurance and heterogenity. We present 2 cases which were referred to our clinic as retroperitoneal mass and diagnosed as CD after surgical resection.