Counterion and substrate effects on barrier heights of the hydrolytic kinetic resolution of terminal epoxides catalyzed by Co(III)-salen.
Density functional theory (DFT) has been applied to the proposed rate-limiting step of the hydrolytic kinetic resolution (HKR) of terminal epoxides as catalyzed by Co-salen-X (X = counterion) in order to resolve questions surrounding the mechanism. The present results indicate that the bimetallic mechanism proposed by Jacobsen shows nonadditive, cooperative catalysis with a larger reduction in barrier height than the sum of the barrier height reductions from the two monometallic reaction pathways. We computed barrier heights for the reaction using several counterions (chloride, acetate, tosylate, and hydroxide). For the three counterions that are experimentally active (chloride, acetate, and tosylate) the barrier heights are 35, 38, and 34 kJ mol(-1), respectively, while for hydroxide it is 48 kJ mol(-1). The similarity of the barrier heights for chloride, acetate, and tosylate is in agreement with their similar peak reaction rates. The finding that Co-salen-X with these counterions leads to rather different overall reaction profiles suggests that they have quite different rates of reaction with epoxide to form the activated Co-salen-OH required for the bimetallic mechanism. Co-salen-OH is inactive as the sole catalyst for HKR, and this inactivity is ascribed to its larger barrier height for the ring-opening step, rather than to any inability to activate epoxide. Barrier heights were also computed using propylene oxide, 1-hexene oxide, and epichlorohydrin; propylene oxide and 1-hexene oxide have similar barrier heights, 35.5 and 33.2 kJ mol(-1), respectively, and epichlorohydrin has a significantly lower barrier height of 18.8 kJ mol(-1), which is qualitatively consistent with experiments showing faster reactions for epicholorohydrin than propylene oxide when catalyzed by Co-salen-OAc.


[Recommendations for sports participation in patients with arrhythmia].
Nowadays, sports are a wonderful mean for social success, and the high-level athlete is the symbol of a perfect hygiene of life. Despite this, the occurrence of unexplained sudden death (SD) is not exceptional, especially during training and competition. In this context, it is important to intensify medical controls for these athletes, especially in a very early phase, in order to detect subjects at risk. In case of detection of a cardiac disease prone to cardiovascular or arrhythmic event, the practice of any high-level sportive activity or even any sustained sportive activity must be forbidden without hesitation, with the aim of protecting these subjects. Even though a total interdiction of sports practice can be a tough decision to be accepted, it should prevail on the dramatic consequences of sudden death. Physicians' responsibility issues in the screening and management of competition or leisure-time sportsmen are of high importance since in case of sudden death, the physician and the medical community liabilities can be considered. As a consequence, the medical community set up recommendations on the screening, treatment and even interdiction of sportive activity for athletes, which should also be applied to leisure-time sportsmen. In the first part of this article, the different causes (especially the arrhythmia-related) of sudden death occurring in sportsmen are reviewed. In the second part, the recommendations on practice of high-level sports in case of arrhythmia or genetic arrhythmic cardiac disease are summarized.


Tough times at the top: Occupational status predicts changes in job satisfaction in times of crisis.
How do individuals with a higher versus lower occupational status experience major, unexpected changes to their work life? The COVID-19 pandemic has disrupted most areas of work life and, thus, provides a unique opportunity to examine changes in work attitudes in response to a worldwide crisis. We predict that individuals with higher, but not with lower occupational status showed a decline in job satisfaction during the early stages of the COVID-19 pandemic in Germany (1st lockdown; March to May 2020), with subsequent recovery to initial job satisfaction levels. Based on role theory and social-psychological theories of hierarchical differentiation, we argue that, due to the profound work-related changes, individuals with higher (vs. lower) occupational status are more negatively affected in realizing their work goals and, thus, experience decreasing levels of job satisfaction. To test these predictions, we investigated trajectories of job satisfaction between December 2019 and August 2020 (7 measurement waves; N = 1583). Results of piece-wise growth curve models showed that individuals with higher occupational status showed a steeper decline in job satisfaction (followed by recovery) over time, whereas individuals with medium and lower occupational status did not experience a significant change in job satisfaction. In addition, we show that the decline in job satisfaction is moderated by perceived constraints at work associated with the pandemic among individuals with higher occupational status. Overall, these findings contribute to our understanding of the link between occupational status and job satisfaction in times of crisis.


Organic acids pretreatment effect on Rosa bourbonia phyto-biomass for removal of Pb(II) and Cu(II) from aqueous media.
The sorption of Pb(II) and Cu(II) form aqueous media by Rosa bourbonia waste phyto-biomass (RBWPB) pretreated with organic acids was investigated as a function of biosorbent dosage, initial metal ions concentration and contact time. The equilibrium biosorption data was analyzed using two kinetic models (pseudo-first order and pseudo-second order) and two isotherm models (Langmuir and Freundlich). The RBWPB was successfully applied for sequestration of both heavy metal ions; however, organic acids pretreatments decreased the metal adsorption capacity of RBWPB. The Langmuir model fitted well to the data, and the pseudo-second order kinetic equation could best describe the biosorption kinetics of Pb(II) and Cu(II) metal ions. Gibbs free energy indicated the spontaneous adsorption of Pb(II) and Cu(II) onto RBWPB.


Cobalt chloride decreases EC-SOD expression through intracellular ROS generation and p38-MAPK pathways in COS7 cells.
It is known that cells suffer a chronic hypoxic condition during the development of proximal tubulointerstitial disease. However, it is accepted that extracellular-superoxide dismutase (EC-SOD) protects the cells from oxidative stress. The purpose of this study was to elucidate the regulation of EC-SOD expression in cells under hypoxia. The results show that the expressions of EC-SOD mRNA and protein in cobalt chloride (CoCl(2))-treated COS7 cells decreased in a dose- and time-dependent manner, whereas the expressions of other SOD isoforms (Cu/Zn-SOD and Mn-SOD) were not changed. The down-regulation of EC-SOD mRNA was suppressed by pre-treatment with the antioxidant trolox and the p38 mitogen-activated protein kinase (p38-MAPK) inhibitor SB203580. It is concluded that the expression of EC-SOD is decreased through ROS and p38-MAPK signalling cascades and that the down-regulation of EC-SOD leads to a decrease in the resistance to oxidative stress of COS7 cells under hypoxia induced by CoCl(2).


Caring for the foot mobile: holistic foot and nail management.
Proper foot and nail management is essential to maintaining a good quality of life and remaining independent as we age. Basic and ongoing care of the feet and proper shoe selection are critical not only for preventing diabetic foot problems, but also for helping to lead productive, balanced lives. When our feet hurt, the rest of our body most likely hurts too. Because proper foot and nail care and education are critical for preventing ulceration and amputation, especially in persons with diabetes, this outcome case study presents a holistic approach to the management of foot and nail problems.


Comparison of transit-time and Doppler ultrasound methods for measurement of flow in aortocoronary bypass grafts during cardiac surgery.
To evaluate the accuracy of flow measurements in aortocoronary bypass grafts with the ultrasound transit-time method, an in vitro and in vivo comparison was carried out. The in vitro comparison with evaluation against both true flow and the ultrasound Doppler method, was carried out with a fresh saphenous vein mounted in a pulsatile flow rig. The two flow probes were placed on the graft 4-5 cm apart to avoid acoustic interference, and blood was pumped through the system at different flow rates. The comparison between the methods showed excellent agreement with a linear correlation coefficient of 0.996, and a mean error of -2.9 ml/min with limits of agreement +/- 13.1 ml/min (+/- 2 SD = 95% of measured differences between these limits). However, against true flow, both methods overestimated flow slightly with mean error 4.4 and 7.3 ml/min for the transit-time and Doppler, respectively. Both methods showed excellent correlation with true flow (correlation coefficient 0.998 for the transit-time and 0.997 for the Doppler method). The in vivo accuracy was evaluated by comparison of the two methods in 9 patients. The two probes were placed on the same saphenous vein grafts 4-5 cm apart, and a total of 34 measurements in 17 grafts were carried out including measurements at baseline and after papaverine injection. The correlation coefficient was 0.990 and linear regression analysis gave the equation: Transit-time flow = 1.00 x Doppler flow + 1.3. In terms of flow, the mean error was 1.5 ml with limits of agreement +/- 17.2 ml.(ABSTRACT TRUNCATED AT 250 WORDS)


RepAM of the Amycolatopsis methanolica integrative element pMEA300 belongs to a novel class of replication initiator proteins.
Accessory genetic elements, such as plasmids and integrative elements, are widespread amongst actinomycetes, but little is known about their functions and mode of replication. The conjugative element pMEA300 from Amycolatopsis methanolica is present mostly in an integrated state at a single specific site in the chromosome, but it can also replicate autonomously. Complete nucleotide sequencing, in combination with deletion studies, has revealed that orfB of pMEA300 is essential for autonomous replication in its host. In this study, it was shown that purified OrfB protein binds specifically to the 3' end of its own coding sequence. Within this short sequence, a putative hairpin structure is located, which contains several direct and inverted repeats, and a nucleotide stretch that resembles the nicking site of the pC194 family of rolling circle replicating plasmids. Additional binding studies revealed that OrfB binds to an 8 bp inverted repeat that occurs three times within the hairpin structure. The data presented show that OrfB is the replication initiator (Rep) protein of pMEA300, and is therefore termed RepAM. Surprisingly, RepAM lacks significant sequence similarity with known prokaryotic Rep proteins, but it is highly similar to a number of yet uncharacterized ORFs that are located on integrative and conjugative elements of other actinomycetes. It is concluded that RepAM and its homologues are members of a novel class of Rep proteins.


A twelve-gene signature for survival prediction in malignant melanoma patients.
Melanoma is defined as a highly mutational heterogeneous disease containing numerous alternations of the molecule. However, due to the phenotypically and genetically heterogeneity of malignant melanoma, conventional clinical characteristics remain restricted or limited in the ability to accurately predict individual outcomes and survival. This study aimed to establish an accurate gene expression signature to predict melanoma prognosis. In this study, we established an RNA sequencing-based 12-gene signature data of melanoma patients obtained from 2 independent databases: the Cancer Genome Atlas (TCGA) database and the Gene Expression Omnibus (GEO) database. We evaluated the quality of each gene to predict survival conditions in each database by employing univariate and multivariate regression models. A prognostic risk score based on a prognostic signature was determined. This prognostic gene signature further classified patients into low-risk and high-risk groups. Based on a prognostic signature, a prognostic risk score was determined. This prognostic gene signature further divided the patients into low-risk and high-risk groups. In the chemotherapy and radiotherapy groups of the TCGA cohort and V-raf murine sarcoma viral oncogene homolog B1 (BRAF) expression group in the GEO cohort, patients in the low-risk group had a longer survival duration compared to patients in the high-risk group. Nevertheless, the immunotherapy group in the TCGA database and neuroblastoma RAS viral oncogene homolog (NRAS) expression group in the GEO database had no significant differences in statistics. Moreover, this gene signature was associated with patient prognosis regardless of whether the Breslow depth was greater than or less than 3.75 mm. Stratified gene set enrichment analysis (GSEA) revealed that certain immune-related pathways, such as the T-cell signaling pathway, chemokine signaling pathway, and primary immunodeficiency, were significantly enriched in the low-risk group of both TCGA and GEO cohorts. This information implied the immune-related properties of the 12-gene signature. Our study emphasizes the significance of the gene expression signature in that it may be an indispensable prognostic predictor in melanoma and has great potential for application in personalized treatment.


Computer-Aided Detection for Pancreatic Cancer Diagnosis: Radiological Challenges and Future Directions.
Radiological imaging plays a crucial role in the detection and treatment of pancreatic ductal adenocarcinoma (PDAC). However, there are several challenges associated with the use of these techniques in daily clinical practice. Determination of the presence or absence of cancer using radiological imaging is difficult and requires specific expertise, especially after neoadjuvant therapy. Early detection and characterization of tumors would potentially increase the number of patients who are eligible for curative treatment. Over the last decades, artificial intelligence (AI)-based computer-aided detection (CAD) has rapidly evolved as a means for improving the radiological detection of cancer and the assessment of the extent of disease. Although the results of AI applications seem promising, widespread adoption in clinical practice has not taken place. This narrative review provides an overview of current radiological CAD systems in pancreatic cancer, highlights challenges that are pertinent to clinical practice, and discusses potential solutions for these challenges.


Isocitrate dehydrogenase mutations in diffuse gliomas: clinical and aetiological implications.
The discovery of isocitrate dehydrogenase (IDH) mutations in gliomas is one example of the large impact that next-generation sequencing is having on the understanding of tumour biology and human disease in general. IDH mutations are early and common events in the development of astrocytomas, oligodendrogliomas and oligoastrocytomas. IDH mutations are also found in some myeloid malignancies and soft tissue tumours, but are rare in other malignancies. IDH mutation detection can be incorporated into routine pathology practice via immunohistochemistry and/or standard sequencing techniques and has great diagnostic value. An emerging theme is that IDH mutation status in gliomas is of great prognostic relevance, and there are proposals to include IDH mutation status in the next iteration of the WHO classification of gliomas. The mechanisms of action(s) of mutant IDH are not fully understood, but the understanding is progressing rapidly, and may provide a mechanism to link diverse proneoplastic processes such as oxidative damage and epigenetic dysregulation. There are exciting prospects of novel therapies for glioma patients emerging from the elucidation of these mechanisms. Given the diagnostic and prognostic implications of IDH mutation, and the potential for new therapies, all gliomas should be assessed for IDH mutation status in the future.


Parents of adults with mental retardation living in-home and out-of-home: caregiving burdens and gratifications.
Interviews with parents of adult children with mental retardation were conducted to identify differences in caregiver burdens and gratifications, based on in-home and out-of-home placement. Parental responses indicate that caretakers of adult children with mental retardation are worried and feel responsibility for their care regardless of residential placement of the adult child.


Three cases of localized cutaneous nodular amyloidosis in patients with limited systemic sclerosis and a brief literature review.
Localized cutaneous nodular amyloidosis (LCNA) is the rarest form of localized cutaneous amyloidosis. In patients with LCNA, local plasma cells secrete immunoglobulin light chains called amyloid L via an unknown mechanism. LCNA has been associated with autoimmune connective tissue diseases such as most commonly Sjögren syndrome. A few reported cases of LCNA are associated with limited systemic sclerosis (LSSc). We report three cases of LCNA in patients with LSSc to add to the existing literature, discuss the disease association and proposed pathophysiology, and briefly review the existing information in current literature. It is important to closely follow patients with LCNA to monitor progression to systemic amyloidosis.


Effect of platelet-activating factor-receptor antagonism on endotoxin-induced lung dysfunction in sheep.
To further define the role of platelet-activating factor (PAF) in endotoxin-induced lung dysfunction, we examined the effect of ABT-299, a specific and potent PAF-receptor antagonist, on the response to endotoxemia in six chronically instrumented awake sheep. We administered Escherichia coli endotoxin (0.5 microg/kg) intravenously with or without pretreatment with ABT-299 while monitoring mean pulmonary arterial pressure (Ppa), mean systemic arterial pressure (Psa), dynamic compliance of the lungs (Cdyn), and functional residual capacity (FRC). Endotoxin administration caused pulmonary hypertension, reduced Cdyn, leukopenia, and hypoxemia while having no significant effect on Psa or FRC. Administration of ABT-299 did not affect any of the measured variables at baseline. Pretreatment with ABT-299 attenuated the peak Ppa seen after endotoxin administration but had minimal effects on endotoxin-induced changes in Cdyn, white blood cell count, or alveolar-to-arterial oxygen difference. ABT-299 was shown to completely block the pulmonary hypertension and reduction in Cdyn seen after intravenous administration of exogenous PAF. We conclude that PAF does not play an essential role in the sheep's response to endotoxin.


Manufacture of Clinical-Grade Human Clonal Mesenchymal Stem Cell Products from Single Colony Forming Unit-Derived Colonies Based on the Subfractionation Culturing Method.
Stem cell products derived from mesenchymal stem cells (MSCs) have been widely used in clinical trials, and a few products have been already commercialized. However, the therapeutic effects of clinical-grade MSCs are still controversial owing to mixed results from recent clinical trials. A potential solution to overcome this hurdle may be to use clonal stem cells as the starting cell material to increase the homogeneity of the final stem cell products. We have previously developed an alternative isolation and culture protocol for establishing a population of clonal MSCs (cMSCs) from single colony forming unit (CFU)-derived colonies. In this study, we established a good manufacturing practice (GMP)-compatible procedure for the clinical-grade production of human bone marrow-derived cMSCs based on the subfractionation culturing method. We optimized the culture procedures to expand and obtain a clonal population of final MSC products from single CFU-derived colonies in a GMP facility. The characterization results of the final cMSC products met our preset criteria. Animal toxicity tests were performed in a good laboratory practice facility, and showed no toxicity or tumor formation in vivo. These tests include single injection toxicity, multiple injection toxicity, biodistribution analysis, and tumorigenicity tests in vivo. No chromosomal abnormalities were detected by in situ karyotyping using oligo-fluorescence in situ hydridization (oligo-FISH), providing evidence of genetic stability of the clinical-grade cMSC products. The manufacture and quality control results indicated that our GMP methodology could produce sufficient clonal population of MSC products from a small amount of bone marrow aspirate to treat a number of patients.


Individualizing endpoints in randomized clinical trials to better inform individual patient care: the TARGET proposal.
In practice, critical care practitioners individualize treatments and goals of care for each patient in light of that patient's acute and chronic pathophysiology, as well as their beliefs and values. Yet critical care researchers routinely measure one endpoint for all patients during randomized clinical trials (RCTs), eschewing any such individualization. More recent methodology work has explored the possibility that enrollment criteria in RCTs can be individualized, as can data analysis plans. Here we propose that the specific endpoints of a RCT can be individualized-that is, different patients within a single RCT might have different secondary endpoints measured. If done rigorously and objectively, based on pre-randomization data, such individualization of endpoints may improve the bedside usefulness of information obtained during a RCT, while perhaps also improving the power and efficiency of any RCT. We discuss the theoretical underpinnings of this proposal in light of related innovations in RCT design such as sliding dichotomies. We discuss what a full elaboration of such individualization would require, and outline a pragmatic initial step towards the use of "individualized secondary endpoints" in a large RCT evaluating optimal enteral nutrition targets in the critically ill.


The effect of intrauterine growth retardation on renal function in the first two months of life.
Children born with growth retardation (GR) have a smaller nephron number and are at increased risk for the development of renal disease and hypertension in adult life. Data on the immediate post-natal development of renal function in neonates born with GR are limited and data on the effects of aminoglycosides (AGs) on renal function in these infants are lacking. This was a prospective study of 81 preterm neonates with a mean gestational age of 32.5 weeks, 40 born with GR (small for gestational age, SGA) and 41 without GR (appropriate for gestational age, AGA). The infants were classified into 4 groups. Groups A (n = 21) and B (n = 20) consisted of AGA and SGA neonates, respectively, who received AGs, and groups C (n = 20) and D (n = 20) of AGA and SGA neonates, respectively, who did not receive AG treatment. Indices of renal function were: serum creatinine (SeCr), the fractional excretion of sodium (FENa), potassium (FEK), phosphorus (FEP), magnesium and uric acid (FEUA), the urinary calcium/creatinine ratio and the transtubular potassium gradient (TTKG). No differences were observed in the parameters examined between SGA and AGA neonates who did not receive AGs. Conversely, SGA infants who received AGs after birth (group B) exhibited higher values of SeCr 2 months later. Specifically, their mean +/- SD value of SeCr (micromol/l) was 42 +/- 05 compared with 33 +/- 08 in group D, 35 +/- 04 in group A and 33 +/- 04 in group C (P < 0.01). These infants also had significantly higher values of TTKG than SGA infants without AG treatment (22 +/- 9 vs 13 +/- 3 in group D) and FEUA (60 +/- 23 vs 35 +/- 14 in group D). Their FENa and FEP were also inappropriately high despite having lower serum levels of Na and P. Preterm SGA infants who had no need of AG treatment after birth have similar renal functional maturation than AGA preterm infants at 2 months of life, but preterm SGA infants who received AGs had indications of impaired glomerular and tubular function at this age.


The ability of seeds to float with water currents contributes to the invasion success of Impatiens balfourii and I. glandulifera.
Two alien species in Europe, Impatiens glandulifera and I. balfourii, are closely related, have similar growth rates and reproductive capacities, and are very attractive to pollinators. Nevertheless, only I. glandulifera is a highly invasive alien species in Europe, while I. balfourii is non-invasive. We assumed that the varying levels of invasiveness are driven by differences in the floating ability of their seeds, which may determine the invasion success of riparian alien plants, such as the Impatiens species. By mimicking two types of aquatic conditions, we determined seed floating ability for each species from younger and older populations. We also analyzed four seed traits: seed viability, surface, shape and coat structure. Seeds of the non-invasive I. balfourii float less well than seeds of the invasive I. glandulifera. We also found that the seeds of I. balfourii from the younger population have a higher floating ability in comparison with that of the seeds from the older population. The results for I. glandulifera were the opposite, with decreased floating ability in the younger population. These differences were associated with seed surface, shape and coat structure. These results indicate that the floating ability of I. balfourii seeds may increase over time following its introduction into a given area, while in the case of I. glandulifera, this ability may gradually decrease. Therefore, the former species, currently regarded as a poor disperser, has the potential to become invasive in the future, whereas the latter does not seem to benefit from further investments in the floating ability of its seeds.


[The Dutch College of General Practitioners' 'Irritable bowel syndrome' standard; reaction from the field of gastroenterology].
The definition of 'irritable bowel syndrome' (IBS) as given by the Dutch College of General Practitioners' standard may be too broad to define IBS. Coeliac disease should be added to the differential diagnosis of IBS. Attention should be given to subjects with an increased familial risk of colorectal cancer. Based on the results of thorough investigations, the role of disturbed motility, visceral hyperalgesia, modified cerebrovisceral perception, induction by infectious diseases and contributing psychological factors should be recognised as pathophysiologically relevant for IBS. When treating patients, making a positive diagnosis of IBS, explaining possible pathophysiological factors and exploring contributing factors is much more important than simply giving reassurance. Medical therapy is of limited value, but may relieve symptoms in selected patients.


HiPS-Endothelial Cells Acquire Cardiac Endothelial Phenotype in Co-culture With hiPS-Cardiomyocytes.
Cell-cell interactions are crucial for organ development and function. In the heart, endothelial cells engage in bidirectional communication with cardiomyocytes regulating cardiac development and growth. We aimed to elucidate the organotypic development of cardiac endothelial cells and cardiomyocyte and endothelial cell crosstalk using human induced pluripotent stem cells (hiPSC). Single-cell RNA sequencing was performed with hiPSC-derived cardiomyocytes (hiPS-CMs) and endothelial cells (hiPS-ECs) in mono- and co-culture. The presence of hiPS-CMs led to increased expression of transcripts related to vascular development and maturation, cardiac development, as well as cardiac endothelial cell and endocardium-specific genes in hiPS-ECs. Interestingly, co-culture induced the expression of cardiomyocyte myofibrillar genes and MYL7 and MYL4 protein expression was detected in hiPS-ECs. Major regulators of BMP- and Notch-signaling pathways were induced in both cell types in co-culture. These results reflect the findings from animal studies and extend them to human endothelial cells, demonstrating the importance of EC-CM interactions during development.


[Validation of the general guidelines of communication between the nurse and the blind].
This quantitative study, conducted between 2008 April and 2009 March in Fortaleza-CE, Brazil, aimed to validate the general guidelines of the communication of the nurse with the blind. Thirty nurses and 30 blinds, divided into control and experimental groups, participated in the study. Thirty nursing consultations were videotaped and analyzed by experts. In contrast to the control group, the experimental group showed good and excellent performance in all guidelines for verbal and non-verbal communication with the blinds. The study results point to the urgency of adopting the teaching of these general guidelines for communicating with the blind in nursing courses, in addition to training nurses in caring for the blind people.


[Vaccinations in patients with autoimmune diseases].
The number of individuals with autoimmune diseases treated with immunosuppressive drugs is increasing steadily. The variety of immunosuppressive drugs and in particular biological therapies is also rising. The autoimmune disease itself as well as the immunosuppressive therapy increases the risk of infection in this population. Particularly the risk of vaccine-preventable infections is elevated. Thus, preventing infections by the means of vaccination is of utmost importance. The Division of Infectious Diseases of the Epidemiology, Biostatistics and Prevention Institute, University of Zurich, performed a literature search on the topic of vaccinations in patients with autoimmune diseases upon request by the Swiss Federal Commission for Vaccination Issues. Overall, data are scarce. The following main points were retrieved from the literature: Inactivated vaccines are safe, but their immunogenicity may be reduced under immunosuppressive therapy. In addition to the generally recommended basic vaccinations, specific vaccinations, such as influenza and pneumococcal vaccination are indicated in these patient groups. Live vaccines are generally contraindicated under immunosuppressive therapy due to safety concerns. However, specific exceptions apply. Furthermore, certain time intervals for the administration of live vaccines after pausing or ceasing an immunosuppressive therapy should be respected.


Bone defect repair with tissue-engineered cartilage.
We tested the efficacy of a new approach for the tissue-engineered growth of cartilage developed in our laboratory in repairing surgically created bone defects in the craniums of rats. Large cranial defects were created bilaterally in the frontoparietotemporal bones of athymic nude rats (n = 10). There was gross evidence of new cartilage in 8 of 10 experimental defects that had been filled with a synthetic biocompatible, biodegradable polymer template that had been seeded in vitro with freshly isolated chondrocytes. The control defects, filled with either nothing at all or a polymer template without chondrocytes, showed no evidence of cartilaginous repair (0 of 10). Statistical analysis using McNemar's test with pooled samples showed significant differences between the two groups (p < 0.05). Prior reports concerning the biologic repair of bony defects involved stimulation of adjacent mesenchymal tissue and resulted in ingrowth of new bone. To our knowledge, this is the first report of structural cartilaginous repair of a bony defect with matrix secreted by implanted chondrocytes.


Protein extraction and activity in reverse micelles of a nonionic detergent.
We describe, for the first time, the ability of a polyoxyethylene sorbitan trioleate-isopropanol microemulsion in hexane to solubilize pure proteins. The dependences of cytochrome c extraction and buffer solubilization by the reverse micellar system on ionic strength of the aqueous phase, detergent concentration, and cosurfactant concentration result in increased extraction. In addition, subtilisin (a serine protease) is shown to be active in this microemulsion. Further the activity of the enzyme can be regulated by the water content of the micelles, enabling control of enzyme activity by "solvent engineering."


Vascular Endothelial Growth Factor as Potential Biomarker for COVID-19 Severity.
COVID-19 is characterized by immunological responses to viral replication and coherent with endothelitis, microvascular disturbance of lung vasculature and coagulopathy. Vascular Endothelial Growth Factor (VEGF) is a proangiogenic mediator regulating endothelial changes. It is induced by proinflammatory signaling and hypoxia. We sought to determine whether VEGF levels differ between SARS-CoV-2-positive patients of different disease severity and whether VEGF might be useful in risk stratification. After retrospective screening of all SARS-CoV-2-positive patients treated in Unfallkrankenhaus Berlin in 2020, we included those with documented VEGF measurement. We extracted laboratory values and clinical parameters. An exploratory data analysis was performed to detect possible relations between VEGF level and clinical disease features. We included 167 SARS-CoV-2-positive patients of which 139 suffered from COVID-19. Seventy-one of the COVID-19 patients had to be treated in the intensive care unit (ICU), those patients exhibited higher VEGF levels than those being admitted to normal wards (535 vs 279 pg/L, P < .001). APACHE-2 (Acute Physiology And Chronic Health Evaluation Score) correlated with mortality and patients with high values showed higher VEGF concentrations on admission (456 vs 875 pg/L, p = 0.006). Receiver operating characteristic analytic revealed that the occurrence of organ dysfunctions like acute respiratory distress syndrome (ARDS), shock, or acute kidney injury could be predicted by VEGF. It was significantly higher in patients who later died compared to survivors (637 vs 389 pg/mL, P = 0.041) and predicted mortality with same accuracy as established markers. In our cohort, association of VEGF above 277 pg/L on admission with risk of ARDS could be confirmed in logistic regression adjusting for possible confounding factors (odds ratio 3.1, 95% confidence interval: 1.34-7.7). Even though there are several limitations to this retrospective study it revealed that in COVID-19 patients VEGF can contribute to the prediction of necessity of ICU, mortality and the prediction of ARDS, kidney injury or shock. Its use in risk stratification and potential pathogenetic involvement should be further investigated.


P4HA1 Down-Regulation Inhibits Glioma Invasiveness by Promoting M1 Microglia Polarization.
Polarization of microglia cells in the glioma microenvironment is closely related to the malignant progression and invasion of gliomas. Prolyl 4-hydroxylase subunit α1 (P4HA1) is the rate-limiting subunit of prolyl 4-hydroxylase (P4H). In previous studies, we showed that P4HA1 could promote the proliferation, migration, and invasion of glioma cells, but the specific mechanisms through which this occurs have not been fully elucidated. Interactions between glioma and microglia cells were analyzed using bioinformatics. Then, co-culture models were used to obtain conditioned media. To characterize microglial cell polarization, we used PCR and immunofluorescence. Proliferation and invasion assays were used to explore the biological behavior of glioma cells affected by microglia. Finally, marker expression was detected using immunohistochemistry in glioblastoma multiform (GBM) specimens. Knockdown of P4HA1 resulted in reduced chemotaxis of microglia toward GBM cells and increased polarization of microglia toward the M1 phenotype. The changed microglial polarization state, in turn, inhibited the proliferation and invasion of GBM cells. Moreover, in GBM tissue specimens, the P4HA1 expression level is negatively correlated with that of the CD86 microglia M1-specific marker. Our results show that P4HA1 promotes immunosuppressive microenvironment formation by cross-talk between GBM and microglia cells and indirectly increases the aggressiveness of GBM.


Sterol methylation in Saccharomyces cerevisiae.
Various nystatin-resistant mutants defective in S-adenosylmethionine: delta 24-sterol-C-methyltransferase (EC 220.127.116.11) were shown to possess alleles of the same gene, erg6. The genetic map location of erg6 was shown to be close to trp1 on chromosome 4. Despite the single locus for erg6, S-adenosylmethionine: delta 24-sterol-C-methyltransferase enzyme activity was found in three separate fractions: mitochondria, microsomes, and the "floating lipid layer." The amount of activity in each fraction could be manipulated by assay conditions. The lipids and lipid synthesis of mutants of Saccharomyces cerevisiae defective in the delta 24-sterol-C-methyltransferase were compared with a C5(6) desaturase mutant and parental wild types. No ergosterol (C28 sterol) could be detected in whole-cell sterol extracts of the erg6 mutants, the limits of detection being less than 10(-11) mol of ergosterol per 10(8) cells. The distribution of accumulated sterols by these mutants varied with growth phase and between free and esterified fractions. The steryl ester concentrations of the mutants were eight times higher than those of the wild type from exponential growth samples. However, the concentration of the ester accumulated by the mutants was not as great in stationary-phase cells. Whereas the head group phospholipid composition was the same between parental and mutant strains, strain-dependent changes in fatty acids were observed, most notably a 40% increase in the oleic acid content of phosphatidylethanolamine of one erg6 mutant, JR5.


Action of antischistosomal drugs, hycanthone and its analog 1A-4 N-oxide, on viral interferon induction.
Two antischistosomal drugs, hycanthone and its indazole analog 1A-4 N-soxide, of which the former is a potent mutagen, differed markedly in their ability to depress interferon induction by influenza virus in rhesus monkey kidney (LLC-MK2) cell cultures. At the concentration of 10 micron/ml, hycanthone reduced interferon yields as much as 73%; the same quantity of analog had no adverse effect on interferon induction. Pretreatment of cell cultures with the analog negated the inhibitory activity of hycanthone on viral interferon induction; however, this did not occur when the sequence was reversed. Interferon-mediated antiviral cellular resistance was not affected when cell cultures were pretreated with either hycanthone or 1A-4 N-oxide. A possible association may exist between the mutagenicity of antischistosomal drugs and their ability to affect interferon synthesis.


Decrease of total antioxidant capacity during coronary artery bypass surgery.
Cardiac surgery induces an oxidative stress, which may lead to impairment of cardiac function. In this study, we aimed to measure the changes of oxidative and antioxidative status of patients undergoing coronary artery bypass surgery (CABG). We studied 79 patients who underwent CABG with and without cardiopulmonary bypass (CPB). Of the 79 patients, 39 had CPB and 40 did not. Blood samples were drawn before, during, and after the surgery. Antioxidant status was evaluated by measuring total antioxidant capacity (TAC), and oxidative status was evaluated by measuring total peroxide (TP) levels and oxidative stress index (OSI). TP and OSI levels increased, while TAC decreased progressively after the beginning of surgery, for all patients. There were negative correlations between TAC levels and aortic cross-clamping period and anastomosis time ( r = -0.553, p < 0.001 and r = -0.500, p < 0.001, respectively). In addition, there was a positive correlation between TAC and ejection fraction (r = 0.647, p < 0.001). During CABG, oxidant and OSI levels significantly increase and TAC significantly decreases. This situation is influenced by long CPB and anastomosis time, and also by low ventricular ejection fraction. We concluded that the patients who undergo CABG are exposed to potent oxidative stress that impairs their TAC. We speculate that supplementation with antioxidant vitamins such as vitamins C and E may be beneficial for patients undergoing CABG.


Pyruvate kinase M1 interacts with A-Raf and inhibits endoplasmic reticulum stress-induced apoptosis by activating MEK1/ERK pathway in mouse insulinoma cells.
Apoptotic death of pancreatic β cells is a major cause of type 2 diabetes mellitus (T2D) progression. Two isoforms of pyruvate kinase, PKM1 and PKM2, have been reported to participate in cell death in several cell types; however, little is known about their causal pathways in pancreatic β-cell death. We examined whether the suppression of PKM1 or PKM2 affects endoplasmic reticulum (ER) stress-induced apoptosis in a pancreatic β-cell line, MIN6, and Beta-TC-6 and found that knockdown of PKM1, but not of PKM2, leads to the induction of ER stress-induced apoptosis in these cells. We also investigated the mechanism by which PKM1 inhibits ER stress-induced apoptosis. We confirmed that PKM1 interacts with A-Raf, an upstream regulator of the MEK/ERK pathway, and that this interaction contributes to MEK1 phosphorylation by A-Raf. PKM1 knockdown suppresses the phosphorylation of MEK, ERK, and caspase-9 (Thr125), which is phosphorylated by the MEK/ERK pathway, thereby inhibiting the cleavage and activation of caspase-9. Thus, PKM1 knockdown activates the caspase-9/caspase-3 pathway under ER stress conditions and leads to apoptosis.


A positional isotope exchange study of the argininosuccinate lyase reaction.
15N nuclear magnetic resonance spectroscopy was used to follow the positional isotope exchange reaction of bovine liver argininosuccinate lyase. The enzyme was shown to catalyze the N-3-N-2 positional nitrogen exchange in [3-15N]argininosuccinate in the presence of excess arginase. The ratio of the positional isotope exchange rate and the rate for net substrate turnover is less than 0.15 at low levels of fumarate but increases to a limiting value of 1.8 at high fumarate. These data have been interpreted to mean that the dissociation of fumarate and arginine from the ternary enzyme complex is random although fumarate is released at least an order of magnitude faster than is arginine from this complex. The rate constant for the release of fumarate from enzyme-arginine-fumarate is at least 6 times faster than the turnover number of the reverse reaction of argininosuccinate lyase. The lower limit for the release of arginine from this same complex is 0.5.


Resistance Training With Partial Blood Flow Restriction in a 99-Year-Old Individual: A Case Report.
In aging populations for which the use of high loads is contraindicated, low load resistance training associated with blood flow restriction (RT-BFR) is an alternative strategy to induce muscle mass gains. This study investigates the effects of RT-BFR on muscle mass, muscle function, and quality of life of a 99-year-old patient with knee osteoarthritis and advanced muscle mass deterioration. Training protocol consisted of 24 sessions of a unilateral free-weight knee extension exercise associated with partial blood flow restriction through a manometer cuff set at 50% of complete vascular occlusion pressure. We evaluated: cross-sectional area (CSA) and thickness (MT) of the vastus lateralis muscle by ultrasound; function through the Timed Up and Go (TUG) test; and quality of life (QoL) by the WHOQOL-bref, WHOQOL-OLD and WOMAC questionnaires. All tests were performed prior to the training period (Pre) and after the 12th (Mid) and 24th (Post) sessions. Changes were considered significant if higher than 2 times the measurement's coefficient of variation (CV). After 24 sessions, there was an increase of 12% in CSA and 8% in MT. Questionnaires scores and TUG values worsened from Pre to Mid and returned in Post. We consider RT-BFR a viable and effective strategy to promote muscle mass gains in nonagenarians and delay the decline in functionality and QoL associated with aging.


Structure of a DNA-bisdaunomycin complex.
The application of detailed structural data bases has now culminated in the successful design of a new generation of bisanthracyclines that form ultratight DNA complexes [Chaires, J. B., Leng, F., Przewloka, T., Fokt, I., Ling, Y. H., Perez-Soler, R., & Priebe, W. (1997) J. Med. Chem. 40, 261-266]. Daunomycin dimers were designed to bind to DNA in complexes resembling those of monomers intercalated at adjacent sites. The goal of the work described here was to determine, with X-ray crystallography, if a potent member of this newly designed and synthesized class of bisanthracyclines (WP631) binds as intended. WP631 is composed of two daunomycin molecules, linked N3' to N3' by a xylyl group. We have solved the 2.2 A X-ray crystal structure of a complex of WP631 bound to [d(CGATCG)]2. We demonstrate, on a detailed molecular level, that the WP631 design strategy is a success. The structures of WP631 and two daunomycin molecules bound to [d(CGATCG)]2 provide the unprecedented opportunity for detailed comparison of mono- and bis-intercalated complexes of the same chromophore, allowing us to distinguish effects of mono-intercalation from those of bis-intercalation. Differences are focused primarily in the centers of the complexes. DNA unwinding and other helical distortions propagate more efficiently to the center of the WP631 complex than to the center of the daunomycin complex.


Targeted Drugs as Maintenance Therapy after Autologous Stem Cell Transplantation in Patients with Mantle Cell Lymphoma.
The treatment landscape for mantle cell lymphoma (MCL) is rapidly evolving toward the incorporation of novel and biologically targeted pharmaceuticals with improved disease activity and gentler toxicity profiles compared with conventional chemotherapeutics. Upfront intensive treatment of MCL includes autologous stem cell transplantation (SCT) consolidation aimed at deepening and lengthening disease remission, but subsequent relapse occurs. Maintenance therapy after autologous SCT in patients with MCL in remission features lower-intensity treatments given over extended periods to improve disease outcomes. Targeted drugs are a natural fit for this space, and are the focus of considerable clinical investigation. This review summarizes recent advances in the field and their potential impact on treatment practices for MCL.


Selective activation of 5'-deoxy-5-fluorouridine by tumor cells as a basis for an improved therapeutic index.
The intracellular metabolism of a new fluoropyrimidine, 5'-deoxy-5-fluorouridine (5'-dFUrd), was compared with the metabolism of 5-fluorouracil (FUra), 5-fluorouridine (FUrd), and 5-fluoro-2'-deoxyuridine (FdUrd) in freshly isolated bone marrow cells and Ehrlich ascites tumor cells. Following exposure to tumor cells, all four fluoropyrimidines were metabolized to identical products (i.e., FUra, 5-fluorouridine 5'-monophosphate, 5-fluorouridine 5'-diphosphate, 5-fluorouridine 5'-triphosphate, and 5-fluoro-2'-deoxyuridine 5'-monophosphate), all produced an incorporation of FUra into RNA (FUd greater than FUra greater than FdUrd greater than 5'-dFUrd), and all completely inhibited thymidylate synthetase activity by 1 hr. However, in bone marrow cells, very different patterns were observed. 5'-dFUrd accumulated in the cells, but there were no measurable metabolism, no incorporation of FUra into RNA, and no inhibition of thymidylate synthetase activity. In contrast, both FUra and FUrd were metabolized and produced an incorporation of FUra into RNA (2.7 pmol FUra per micrograms RNA and 4.8 pmol FUra per micrograms RNA at 2 hr, respectively) in bone marrow. Only a minor inhibition of thymidylate synthetase activity was detected. FdUrd also was metabolized by bone marrow cells, produced a low level of FUra incorporation into RNA (0.23 pmol FUra per micrograms RNA at 2 hr), and produced a complete inhibition of thymidylate synthetase activity. Since 5'-dFUrd is not directly cytotoxic itself, its superior therapeutic index compared to other fluoropyrimidines may largely reflect the selective activation of 5'-dFUrd by sensitive tumor cells as opposed to bone marrow cells, which can activate FUra, FUrd, and FdUrd.


Global analysis of hematopoietic and vascular endothelial gene expression by tissue specific microarray profiling in zebrafish.
In this study, we utilize fluorescent activated cell sorting (FACS) of cells from transgenic zebrafish coupled with microarray analysis to globally analyze expression of cell type specific genes. We find that it is possible to isolate cell populations from Tg(fli1:egfp)(y1) zebrafish embryos that are enriched in vascular, hematopoietic and pharyngeal arch cell types. Microarray analysis of GFP+ versus GFP- cells isolated from Tg(fli1:egfp)(y1) embryos identifies genes expressed in hematopoietic, vascular and pharyngeal arch tissue, consistent with the expression of the fli1:egfp transgene in these cell types. Comparison of expression profiles from GFP+ cells isolated from embryos at two different time points reveals that genes expressed in different fli1+ cell types display distinct temporal expression profiles. We also demonstrate the utility of this approach for gene discovery by identifying numerous previously uncharacterized genes that we find are expressed in fli1:egfp-positive cells, including new markers of blood, endothelial and pharyngeal arch cell types. In parallel, we have developed a database to allow easy access to both our microarray and in situ results. Our results demonstrate that this is a robust approach for identification of cell type specific genes as well as for global analysis of cell type specific gene expression in zebrafish embryos.


Feeding ecology of two lanternfishes Diaphus garmani and Diaphus chrysorhynchus.
The feeding ecology of two dominant lanternfishes Diaphus garmani and Diaphus chrysorhynchus was studied in the continental slope region of the East China Sea, off western Kyushu (31-33° N; 128-130° E). Stomach contents of D. garmani were composed mainly of crustacean zooplankton, such as copepods, euphausiids, decapod larvae and amphipods, and also of appendicularians. Stomach contents of D. chrysorhynchus were composed mainly of crustacean zooplankton, cephalopods and fishes. Diel changes in stomach fullness indicated that D. garmani fed more actively at night than in the day. On the other hand, although feeding activity of D. chrysorhynchus did not change drastically between day and night, it tended to feed on large prey items in the benthopelagic zone during the day and on zooplankton in the epipelagic zone at night. Daily rations of food were estimated to be 2·54% of body dry mass for D. garmani, and 2·38% of body dry mass for D. chrysorhynchus.


Hsp90 inhibitor induces KG-1a cell differentiation and apoptosis via Akt/NF-κB signaling.
Heat-shock protein 90 (Hsp 90) acts as a molecular chaperone that maintains protein stability and regulates cell proliferation, survival, differentiation and apoptosis. The present study investigated the effect of Hsp90 inhibition on human acute myeloid leukemia (AML) cells using the novel small-molecule inhibitor SNX-2112. We found that SNX-2112 more potently inhibited KG-1a cell growth than the classical Hsp90 inhibitor 17-(2-dimethylaminoethyl)amino‑17-demethoxygeldanamycin as determined by CCK-8 assay. Flow cytometry was used to examine the cell cycle, differentiation, and apoptosis, and western blotting and qRT-PCR were used to analyze the underlying mechanism. The results revealed that low concentrations of SNX-2112 arrested the cells in the G2/M phase and induced their differentiation and apoptosis, possibly by suppressing Akt and inhibitor of κB kinase, a component of the nuclear factor (NF)-κB signaling pathway. We also found that SNX-2112 increased the expression of the differentiation transcription factors PU.1 and CCAAT‑enhancer-binding protein-α. Thus, SNX-2112 induced KG-1a cell differentiation, cell cycle arrest and apoptosis via modulation of Akt and NF-κB signaling, suggesting that it is a promising therapeutic agent for the treatment of AML.


WNT2 and human gastrointestinal cancer (review).
WNT2 gene on human chromosome 7q31 is a paralog of the WNT2B gene on human chromosome 1p13. Rat Wnt2 gene was identified within rat genome draft sequence AC095247.4. Human WNT2 showed 96.4% total-amino-acid identity with rat Wnt2, 96.1% with mouse Wnt2, 68.6% with zebrafish wnt2, and 67.8% with fugu wnt2. WNT2 is an evolutionarily conserved secreted-type glycoprotein belonging to the WNT family. WNT2 mRNA is expressed in human fetal lung and placenta, but almost undetectable in normal gastrointestinal tract. WNT2 mRNA is frequently up-regulated in human gastric cancer due to tumor-stromal interaction, and WNT2 gene is rarely amplified in human gastric cancer. WNT2 mRNA is also frequently up-regulated in colorectal polyps, primary colorectal cancer of stage A-C, and also in liver metastasis from colorectal cancer. Putative biding sites for estrogen receptor, GATA-1, AP-2, TCF-1, BHLH, MBF-I, p53, and HNF-5 are located within the 5'-flanking region of human WNT2 gene. WNT2 is up-regulated by beta-estradiol in human MCF-7 cells; however, the mechanism of WNT2 up-regulation in most cases of gastrointestinal cancer remains to be elucidated. WNT2 is a tumor marker of gastric and colorectal cancer. Detection of theWNT2 protein in feces by immunohistochemistry or ELISA and WNT2 mRNA in feces by cDNA-PCR or custom-made microarray could be applied for screening of colorectal cancer. Because WNT2 up-regulation leads to carcinogenesis through activation of the WNT-beta-catenin pathway, WNT2 specific antagonist might be applied for chemoprevention or treatment of gastrointestinal cancer. WNT2 gene is one of the targets for pharmacogenomics in the field of oncology.


Obstructive sleep apnea syndrome: An important piece in the puzzle of cardiovascular risk factors.
The obstructive sleep apnea syndrome (OSA) is a clinical entity characterized by recurring episodes of apnea and/or hypopnea during sleep, due to a total or partial collapse, respectively, of the upper airway. This collapse originates a set of pathophysiological changes that determine the appearance of several cardiovascular complications. OSA contributes for the development of hypertension, heart failure, arrhythmias and coronary heart disease. Nowadays it is recognized to be an important public health problem, taking into account not just its repercussions but also its prevalence, since the main risk factor for the disease is obesity, a growing problem worldwide, both in developed and developing countries. The present review summarizes the current knowledge about OSA, as regards its definition, pathophysiology, clinical manifestations, diagnosis, cardiovascular effects and treatment.


Micellar liquid chromatography for lipophilicity determination of new biologically active 1,3-purinodiones.
A series of newly synthesized 1,3-purinodiones with potential anticonvulsant activity, exhibiting affinity to adenosine A(1) and/or A(2A) receptors, were subjected to micellar LC (MLC) with SDS as micelle-forming agent and n-propanol as organic modifier. Two C18 silica-based columns were employed in MLC: a particle one and a monolithic. In parallel, those derivatives were also analyzed in RP-LC on four silica-based columns and on an immobilized artificial membrane column. The correlations between the relevant logarithms of the retention factors of analytes obtained in MLC, immobilized artificial membrane and RP-LC systems on the one hand, and the calculated log P (clog P) and log D values (clog D) on the other, were examined. The level of the correlations of retention data from MLC and RP-LC systems with clog P and clog D obtained is similar but it could be stressed that MLC allows increasing the speed of analysis and using only one mobile phase. Moreover, there is no need of applying an extrapolation procedure in lipophilicity determination. Therefore, the MLC systems, providing chromatographic data in a fast and efficient manner, were demonstrated as promising alternatives to the classical RP-LC systems to estimate the lipophilicity of drugs and drug candidates.


Aortic valve perforation in the setting of Cogan's syndrome.
Cogan's syndrome is a rare disorder characterized by the coexistence of ocular and audio-vestibular manifestations. Systemic manifestations are quite unusual with pan-vasculitis and cardiac involvement reported in the form of aortitis with aortic aneurysm, dissection, or extremely rare aortic valve perforation. Hereby, we report a case of a 56-year-old woman presented with ocular, audio-vestibular, and systemic manifestations with medium-sized vasculitis in the form of multiple splenic artery aneurysms, superior mesenteric artery thrombosis, and cardiovascular involvement in the form of aortic regurgitation due to noncoronary cusp perforation. To the best of our knowledge, this is the second case to report aortic perforation in the setting of Cogan's syndrome.


Response of a chigger, Eutrombicula hirsti (Acari: Trombiculidae) to repellent and toxicant compounds in the laboratory.
Laboratory tests were conducted to compare the response of Eutrombicula hirsti (Sambon) larvae to the compounds dibutylphthalate (DBP), dimethylphthalate (DMP), diethyl methylbenzamide (Deet), benzyl benzoate (BB), and permethrin. The ED50 for E. hirsti of these compounds when applied to filter paper were 0.17 mg/cm2 for DMP, 0.24 mg/cm2 for permethrin, 0.26 mg/cm2 for Deet, 1.42 mg/cm2 for BB, and > 7.21 mg/cm2 for DBP. When impregnated in cloth, DMP, Deet, and BB were highly toxic to larvae at low concentrations, whereas permethrin and DBP were not as rapidly toxic, but were effective.


hmSEEKER: Identification of hmSILAC Doublets in MaxQuant Output Data.
Heavy methyl Stable Isotope Labeling with Amino acids in Cell culture (hmSILAC) is a metabolic labeling strategy employed in proteomics to increase the confidence of global identification of methylated peptides by MS. However, to this day, the automatic and robust identification of heavy and light peak doublets from MS-raw data of hmSILAC experiments is a challenging task, for which the choice of computational methods is very limited. Here, hmSEEKER, a software designed to work downstream of a MaxQuant analysis for in-depth search of MS peak pairs that correspond to light and heavy methyl-peptide within MaxQuant-generated tables is described with good sensitivity and specificity. The software is written in Perl, and its code and user manual are freely available at Bitbucket (https://bit.ly/2scCT9u).


Homoisoflavonoids from Caesalpinia sappan displaying viral neuraminidases inhibition.
In this study, twelve neuraminidase (NA) inhibitory compounds 1-12 were isolated from heartwood of Caesalpinia sappan on the basis of their biological activities against three types of viral NAs. Of isolated homoisoflavonoids, sappanone A (2) showed the most potent NAs inhibitory activities with IC(50) values of 0.7 µM [H1N1], 1.1 µM [H3N2], and 1.0 µM [H9N2], respectively, whereas saturated homoisoflavonoid (3) did not show significantly inhibition. This result revealed that α,β-unsaturated carbonyl group in A-ring was the key requirements for viral NAs inhibitory activity. In our enzyme kinetic study, all NA inhibitors screened were found to be reversible noncompetitive types.


Effect of intermedin1-53 on angiotensin II-induced hypertrophy in neonatal rat ventricular myocytes.
Intermedin (IMD) is coexpressed in the heart with its receptor, which suggests that it may have localized actions as a modulator of cardiac function. The present study was designed to observe the interaction between IMD and cardiac hypertrophy and the possible mechanism involved in the antihypertrophic effects of IMD1-53 in cultured neonatal ventricular myocytes. Myocyte hypertrophy was induced by treating the cells with angiotensin II, and the hypertrophic response was characterized by a significant increase in cell surface area, protein synthesis, and BNP mRNA expression. Our results showed that angiotensin II led to an obvious decrease in the production, secretion, and mRNA expression of IMD and increase receptor activity modifying proteins 1, 3 mRNA expression. Moreover, IMD1-53 inhibited the angiotensin II-induced hypertrophic response and the effects of IMD1-53 were similar to those of equivalent-dose adrenomedullin and could been blocked by H89. Otherwise, in our study, IMD1-53 resulted in dose-dependent increases of cAMP production in cardiomyocytes. Thus, IMD and its receptor system are involved in cardiac hypertrophy, and like adrenomedullin, IMD1-53 exerts an antihypertrophic effect on neonatal cardiomyocytes and the effect can be mediated by the cAMP/PKA pathway.


Pharyngeal symptoms caused by cervical osteophytes.
A case of cervical osteophytes producing the sensation of a 'lump in the throat' is described. This symptom was relieved by excision of the offending osteophytes. Emphasis is placed on the need for surgical correction before progression of degenerative disease makes the procedure technically impossible. Several points concerning the operative and postoperative management are made.


75 years of the Division of Analytical Chemistry of the American Chemical Society.
The Division of Analytical Chemistry is celebrating the 75th anniversary of its founding in 1938. We celebrate the continuing high importance of our discipline for all aspects of chemical science and for its applications in so many aspects of everyday life. We especially celebrate the accomplishments of our fellow analytical chemists through the years, and the impact we have had on the profession. This article is a short history of the Division within the context of the parallel development of our profession and our science.


Right ventricular and tricuspid valve remodeling after bidirectional cavopulmonary anastomosis.
There are few investigations of the changes in tricuspid valve (TV) and right ventricular (RV) morphology following bidirectional cavopulmonary anastomosis (BCPA). The 2-D echocardiograms of 35 children (male, n=23; female, n=12; median age, 6 months; range, 3-10 months) with hypoplastic left heart syndrome, 1 month before and after BCPA performed between 2005 and 2011, were retrospectively reviewed. Patients who underwent TV repair at BCPA were excluded. From the 4-chamber view, the coaptation length, vena contracta width and RV end-diastolic area before and after BCPA were measured and indexed to surface area. The severity of tricuspid regurgitation was graded qualitatively. After BCPA, RV end-diastolic area decreased from 2,951 ± 584 to 2,580 ± 591 mm(2)/m(2) (P<0.001). The coaptation length of the anterior leaflet (8.8 ± 5.8 vs. 11.0 ± 6.2 mm/m(2), P=0.0014) and of the septal leaflet (13.5 ± 5.3 vs. 15.8 ± 5.4mm/m(2), P=0.0072) increased after BCPA. The vena contracta width decreased (5.8 ± 4.9 vs. 4.3 ± 4.2 mm/m(2), P=0.035), although there was no change in tricuspid regurgitation grade after BCPC (1.4 ± 0.7 vs. 1.4 ± 0.9, P=0.234). In children with hypoplastic left heart syndrome after BCPA, the coaptation length of the anterior and septal leaflets of the TV improved concomitantly with vena contracta width and RV end-diastolic area despite unchanged tricuspid regurgitation grade. This suggests that favorable RV and TV remodeling accompanies the reduction in RV volume load following BCPA.


Disruption of Hepatic Mitochondrial Pyruvate and Amino Acid Metabolism Impairs Gluconeogenesis and Endurance Exercise Capacity in Mice.
Exercise robustly increases the glucose demands of skeletal muscle. This demand is met not only by muscle glycogenolysis, but also by accelerated liver glucose production from hepatic glycogenolysis and gluconeogenesis to fuel mechanical work and prevent hypoglycemia during exercise. Hepatic gluconeogenesis during exercise is dependent on highly coordinated responses within and between muscle and liver. Specifically, exercise increases the rate at which gluconeogenic precursors such as pyruvate/lactate or amino acids are delivered from muscle to the liver, extracted by the liver, and channeled into glucose. Herein, we examined the effects of interrupting gluconeogenic efficiency and capacity on exercise performance by deleting hepatic mitochondrial pyruvate carrier 2 (MPC2) and/or alanine transaminase 2 (ALT2) in mice. We found that deletion of MPC2 or ALT2 alone did not significantly affect time to exhaustion or post-exercise glucose concentrations in treadmill exercise tests, but mice lacking both MPC2 and ALT2 in liver (DKO) reached exhaustion faster and exhibited lower circulating glucose during and after exercise. Use of ²H/¹³C metabolic flux analyses demonstrated that DKO mice exhibited lower endogenous glucose production owing to decreased glycogenolysis and gluconeogenesis at rest and during exercise. The decreased gluconeogenesis was accompanied by lower anaplerotic, cataplerotic, and TCA cycle fluxes. Collectively, these findings demonstrate that the transition of the liver to the gluconeogenic mode is critical for preventing hypoglycemia and sustaining performance during exercise. The results also illustrate the need for interorgan crosstalk during exercise as described by the Cahill and Cori cycles.


Acoustic mode coupling induced by shallow water nonlinear internal waves: sensitivity to environmental conditions and space-time scales of internal waves.
While many results have been intuited from numerical simulation studies, the precise connections between shallow-water acoustic variability and the space-time scales of nonlinear internal waves (NLIWs) as well as the background environmental conditions have not been clearly established analytically. Two-dimensional coupled mode propagation through NLIWs is examined using a perturbation series solution in which each order n is associated with nth-order multiple scattering. Importantly, the perturbation solution gives resonance conditions that pick out specific NLIW scales that cause coupling, and seabed attenuation is demonstrated to broaden these resonances, fundamentally changing the coupling behavior at low frequency. Sound-speed inhomogeneities caused by internal solitary waves (ISWs) are primarily considered and the dependence of mode coupling on ISW amplitude, range width, depth structure, location relative to the source, and packet characteristics are delineated as a function of acoustic frequency. In addition, it is seen that significant energy transfer to modes with initially low or zero energy involves at least a second order scattering process. Under moderate scattering conditions, comparisons of first order, single scattering theoretical predictions to direct numerical simulation demonstrate the accuracy of the approach for acoustic frequencies upto 400 Hz and for single as well as multiple ISW wave packets.


[Morphological and functional studies in a case of primary (idiopathic) thrombocythaemia in childhood (author's transl)].
Primary (idiopathic) thrombocythaemia is a rare disease in childhood. Haemorrhagic diathesis and splenomegaly are principal findings besides thrombocytosis. The cases of a nine year old girl with thrombocyte levels between 2.8 and 4.5 X 10(6) microliter is described. The examination in light and electron microscopy revealed various morphological abnormalities in thrombocytes and megakaryocytes. Platelet function was disturbed in various degrees. The impact of functional abnormalities on the haemorrhagic diathesis is discussed. The findings in this child were compared to those of the better characterized picture of primary and secondary thrombozytosis in adulthood. Possible pathogenic relations to myeloproliferative diseases are also discussed.


Comparative clinical trial of bemetizide/triamterene and cyclopenthiazide/potassium chloride combinations in patients with mild to moderate hypertension.
A double-blind trial was carried out to compare the combination of 25 mg bemetizide plus 50 mg triamterene ('Hypertane') and 0.25 mg cyclopenthiazide plus 600 mg potassium chloride ('Navidrex' K) in the treatment of mild to moderate essential hypertension. Two well matched groups of patients were treated for periods of 6 weeks with one or other of the drugs under test. There were 2-week placebo run-in and run-out periods. Blood pressure and laboratory investigations were performed every 2 weeks during the trial period. Both treatments resulted in similar overall statistically significant reductions in blood pressure during the trial. With bemetizide/triamterene, mean lying blood pressure decreased by 11.1/11.2 mmHg and mean standing blood pressure by 15.9/10.3 mmHg; with cyclopenthiazide/potassium chloride the corresponding reductions were 14.9/12.1 mmHg and 9.1/11.7 mmHg. The fact that some of the observed overall reduction seen with both drugs was due to 'placebo effect' is discussed but the clinical importance of overall changes is stressed. There were no significant differences between changes in blood pressure with the two treatments. Biochemical changes were those expected with thiazide diuretics. However, the decrease in potassium and increases in urea and uric acid levels were less with bemetizide/triamterene than with cyclopenthiazide/potassium chloride. Clinical tolerance of both treatments was good.


Chlamydial antibodies in farmers in north-west England.
Because of recent reports of abortion in farmers' wives following infection with ovine strains of Chlamydia psittaci during pregnancy, the distribution of chlamydial antibodies was studied in rural populations in north-west England, where endemic chlamydial infection with abortion is common in sheep. Immunoperoxidase assays with C. trachomatis and ovine C. psittaci showed no significant differences in either the frequency or titres of antibodies between sheep farmers and other types of farmer or non-farming adults living in the same areas. The frequency and titres of antibodies in farmers' wives were no greater than in farmers, and were unrelated to their previous obstetric history or type of farming. Overall, 62/255 (24%) of this rural population had antibody detected by C. trachomatis antigen and only 30/255 (13%) detected by C. psittaci antigen. The possible significance of these findings is discussed. This survey does not suggest that the risk of infection with C. psittaci is especially high in people working with sheep, but the complications following infection during pregnancy deserve the specific instructions that have been given to pregnant women to avoid exposure, especially during lambing, in farming and veterinary work.


Does a patient-held quality-of-life diary benefit patients with inoperable lung cancer?
To examine the effect of weekly completion of a patient-held quality-of-life (QOL) diary in routine oncology practice for palliative care patients. In a pragmatic randomized controlled trial, 115 patients with inoperable lung cancer were randomly assigned to receive either standard care or a structured QOL diary (European Organisation for Research and Treatment of Cancer Quality of Life Questionnaire C30 and the related lung cancer module LC13) that they completed at home each week for 16 weeks. Patients were encouraged to share the QOL information with health professionals involved in their care. Changes in QOL over time (measured by the Functional Assessment of Cancer Therapy-Lung questionnaire and the Palliative Care Quality of Life Index), discussion of patient problems, and satisfaction with communication and general care were assessed at baseline and at 2 and 4 months after baseline. Analysis of QOL indicated a small but consistent difference between patients in the diary group and the standard care group. The diary group had a poorer QOL in many domains. Two different QOL summary scores (total and overall QOL) indicated a statistically significant between-group difference. No effects were found in relation to satisfaction with care, communication, or the discussion of patient problems. The regular completion of a QOL questionnaire without appropriate feedback to health care professionals and without the provision of appropriate support may have a negative impact on inoperable lung cancer patients. Further research should focus on identifying features such as feedback loops that are required for the successful and meaningful use of QOL questionnaires in routine patient care.


Computed tomography-guided biopsy of small lung nodules: diagnostic accuracy and analysis for true negatives.
We evaluated the diagnostic accuracy of computed tomography (CT)-guided transthoracic core needle biopsy (TCNB) for small (≤20-mm) lung nodules and identified predictive factors for true negatives among benign biopsy results. From March 2010 to June 2015, 222 patients with small lung nodules underwent CT-guided TCNB. We retrospectively analysed data regarding technical success, diagnostic accuracy, and predictors of true negatives. The technical success rate was 100%. The TCNB results of the 222 lung nodules included malignancy (n = 136), suspected malignancy (n = 8), specific benign lesion (n = 17), and nonspecific benign lesion (n = 61). The final diagnosis of 222 lung nodules included malignant (n = 160), benign (n = 60), and nondiagnostic lesions (n = 2). The sensitivity, specificity, and overall diagnostic accuracy of CT-guided TCNB for small lung nodules were 90.0%, 100%, and 92.7%, respectively. Pneumothorax and haemoptysis occurred in 23 and 41 patients, respectively. Based on the Cox regression analysis, the significant independent predictive factor for true negatives was a biopsy result of chronic inflammation with fibroplasia. CT-guided TCNB offers high diagnostic accuracy for small lung nodules, and a biopsy result of chronic inflammation with fibroplasia can predict a true-negative result.


Fulminant descending mediastinitis secondary to infectious mononucleosis.
Descending mediastinitis is a rare, life-threatening condition caused by contiguous spread of oropharyngeal or cervical infection into the mediastinum. Infectious mononucleosis generally results in a self-limited illness characterized by fever, pharyngitis and lymphadenopathy. We present an exceptional case of an 18-year-old with infectious mononucleosis complicated by progressive bacterial superinfection and fulminant descending mediastinitis. After resuscitation, broad spectrum antibiotics, critical care support and definitive surgical management, they made a full recovery.


Evidence for sex-specific genetic architectures across a spectrum of human complex traits.
Sex differences are a common feature of human traits; however, the role sex determination plays in human genetic variation remains unclear. The presence of gene-by-sex (GxS) interactions implies that trait genetic architecture differs between men and women. Here, we show that GxS interactions and genetic heterogeneity among sexes are small but common features of a range of high-level complex traits. We analyzed 19 complex traits measured in 54,040 unrelated men and 59,820 unrelated women from the UK Biobank cohort to estimate autosomal genetic correlations and heritability differences between men and women. For 13 of the 19 traits examined, there is evidence that the trait measured is genetically different between males and females. We find that estimates of genetic correlations, based on ~114,000 unrelated individuals and ~19,000 related individuals from the same cohort, are largely consistent. Genetic predictors using a sex-specific model that incorporated GxS interactions led to a relative improvement of up to 4 % (mean 1.4 % across all relevant phenotypes) over those provided by a sex-agnostic model. This further supports the hypothesis of the presence of sexual genetic heterogeneity across high-level phenotypes. The sex-specific environment seems to play a role in changing genotype expression across a range of human complex traits. Further studies of GxS interactions for high-level human traits may shed light on the molecular mechanisms that lead to biological differences between men and women. However, this may be a challenging endeavour due to the likely small effects of the interactions at individual loci.


Advance directives use in acute care hospitals.
It is a federal requirement for health care organizations to provide and obtain information from admitted patients about their rights to self-determination and executing an advance directive. But how compliant are acute care organizations with this law? This article explores the extent to which hospitalized patients in one Southeastern United States organization had advance directive documentation; its correlation with socioeconomic factors, and the success of educating patients and providing care that is consistent with patient's wishes.


Induction of innate immunity by lipid A mimetics increases survival from pneumonic plague.
This study analysed the effect of priming the innate immune system using synthetic lipid A mimetics in a Yersinia pestis murine pulmonary infection model. Two aminoalkyl glucosaminide 4-phosphate (AGP) Toll-like receptor 4 (TLR4) ligands, delivered intranasally, extended time to death or protected against a lethal Y. pestis CO92 challenge. The level of protection was dependent upon the challenge dose of Y. pestis and the timing of AGP therapy. Protection correlated with cytokine induction and a decreased bacterial burden in lung tissue. AGP protection was TLR4-dependent and was not evidenced in transgenic TLR4-deficient mice. AGP therapy augmented with subtherapeutic doses of gentamicin produced dramatically enhanced survival. Combined, these results indicated that AGPs may be useful in protection of immunologically naive individuals against plague and potentially other infectious agents, and that AGP therapy may be used synergistically with other therapies.


Focal lymphocytic infiltration in the adrenal cortex of the elderly: immunohistological analysis of infiltrating lymphocytes.
The incidence of mononuclear cell infiltration in the adrenal cortex was examined in autopsy cases of young and old subjects, and the infiltrating mononuclear cells were immunohistologically characterized by monoclonal antibodies. Histologically, 110 of 174 autopsy cases of persons greater than 60 years (63.2%) were shown to have mononuclear cell infiltration of varying degree within the adrenal cortex, whereas such a lesion was observed in lesser incidence (7.4%) in the 54 younger, control subjects aged less than 49 years. In addition, severely infiltrating lesions in the adrenal cortex were found frequently in the elderly greater than 70 years. Immunohistochemical study revealed that the infiltrating mononuclear cells were mainly composed of CD3+ T cells. The major proportion of CD3+ T cells expressed CD4, whereas CD8+ T cells were less in number. Moreover, a considerable proportion of CD4+ T cells was activated as judged by interleukin 2 receptor expression. These findings indicate that T lymphocytes infiltration in aged human adrenal cortex may represent a pre-clinical manifestation of organ-specific autoimmune adrenalitis which is based on autoimmunity associated with ageing process.


Increasing resistance to penicillin in Streptococcus pneumoniae in southern Sweden.
The susceptibility to penicillin of 6 prevalent pneumococcal types isolated from nasopharynx in 1992 was compared with that of corresponding types from 1980-82. The 6 types or groups, 6, 9, 14, 15, 19 and 23, constituted 78% of consecutive isolates. 19/204 isolates in 1992 were intermediately resistant (MIC 0.12-1.0 mg/l) in comparison with 1/194 from 1980-82 (p < 0.001). Resistant strains (MIC > or = 2 mg/l) were not found. Of group 15, no fewer than 10/31 isolates were intermediately resistant, which may support the clonal origin and spread of penicillin-resistant pneumococci. At least 5.0% of nasopharyngeal isolates are now intermediately resistant to penicillin. This figure is substantially higher than the 2% earlier reported in Sweden.


Linear approximations for swing leg motion during gait.
The applicability of a linear systems analysis of two-dimensional swing leg motion was investigated. Two different linear systems were developed. A linear time-varying system was developed by linearizing the nonlinear equations describing swing leg motion about a set of nominal system and control trajectories. Linear time invariant systems were developed by linearizing about three different fixed limb positions. Simulations of swing leg motion were performed with each of these linear systems. These simulations were compared to previously performed nonlinear simulations of two-dimensional swing leg motion and the actual subject motion. Additionally, a linear system analysis was used to gain some insight into the interdependency of the state variables and controls. It was shown that the linear time varying approximation yielded an accurate representation of limb motion for the thigh and shank but with diminished accuracy for the foot. In contrast, all the linear time invariant systems, if used to simulate more than a quarter of the swing phase, yielded generally inaccurate results for thigh shank and foot motion.


The nonsurgical nature of patients with subarachnoid or intraparenchymal hemorrhage associated with mild traumatic brain injury.
Mild traumatic brain injury (mTBI), as defined by Glasgow Coma Scale (GCS) score of 13 or higher, is a common problem in the United States and worldwide, estimated to affect more than 1 million patients yearly. When associated with intracranial hemorrhage, it is a common reason for neurosurgical consultation and transfer to tertiary care centers. The authors set out to investigate the clinical implications of subarachnoid hemorrhage (SAH) and/or intraparenchymal hemorrhage (IPH) associated with mTBI in hopes of standardization of mTBI clinical care and optimization of resource allocation. The authors performed a retrospective review of 500 consecutively treated patients with mTBI and SAH and/or IPH admitted to a Level I trauma center in Alabama between May 2003 and May 2013. They performed a review of medical records to confirm the diagnosis, determine neurological condition at admission, and assess for episodes of neurological decline or brain injury-related complications including altered mental status, seizures, and hyponatremia. Of the 500 patients for whom data were reviewed, 304 (60.8%) were male and 196 (39.2%) were female. Average age was 46.3 years. Overall, 63 patients (12.6%) had isolated IPH, 411 (82.2%) had isolated SAH, and 26 (5.2%) had radiographic evidence of both IPH and SAH. One hundred forty-five patients (29%) were transferred an average distance of 64.5 miles. The authors identified no patients who experienced neurological worsening during their hospital course. Two patients experienced hyponatremia that required treatment with sodium supplementation. Patients with the constellation of SAH and/or IPH and mTBI do not require neurosurgical consultation, and these findings should not be used as the sole criteria to justify transfer to tertiary referral centers.


[Zinc finger nucleases and their application].
Zinc finger nuclease (ZFN), which is a chimeric fusion structure between a Cys2-His2 zinc-finger protein (ZFP) and the cleavage domain of Fok I endonuclease, can be used to introduce targeted double-stranded breaks (DSBs). ZFN-mediated cleavage leads to mutations when double-stranded breaks are repaired by homologous recombination (HR) or nonhomologous end joining (NHEJ). In recent years, ZFNs are widely used in the fields of genetic research. In this review, the methodology and technical advantages of ZFNs were briefly discussed.


Retinoic acids change gene expression profiles of bovine intramuscular adipocyte differentiation, based on microarray analysis.
Beef marbling is caused by intramuscular deposition and it is an economically important trait in the beef industry. Vitamin A (VA) is an important feed supplement for cattle, but it can hinder marbling if provided in excess. In cattle, VA forms various derivatives such as all-trans retinoic acid (ATRA) and 9-cis retinoic acid (9cRA). Therefore, we investigated the genes involved in bovine intramuscular adipogenesis after VA treatment with ATRA and 9cRA. Differential gene expression levels were validated by microarray analysis in a clonal bovine intramuscular preadipocyte (BIP) cell line derived from the intramuscular adipose tissue of Japanese Black cattle. BIP cells were harvested 6 days after adipogenic stimulation with either 1 μmol/L ATRA, 1 μmol/L 9cRA or non-retinoic acid control. The ATRA- and 9cRA-treated cells exhibited reduced transcription of genes involved in the circulatory system and muscle development compared with the no retinoic acid (RA) treatment. In addition, the ATRA- and 9cRA-treated cells exhibited increased transcription of genes involved in the immune system, protein kinase B signaling and responses to various stimuli. These results demonstrate the lower expression of muscle development in ATRA- and 9cRA-treated BIP cells during adipogenesis.


Transcriptional regulation of the mouse gene encoding the alpha-4 subunit of the GABAA receptor.
The gamma-aminobutyric acid type A receptors (GABAA-Rs) mediate fast inhibitory synaptic transmission in the brain. The alpha4 subunit of the GABAA-R confers distinct pharmacological properties on the receptor and its expression pattern exhibits plasticity in response to physiological and pharmacological stimuli, including withdrawal from progesterone and alcohol. We have analyzed the promoter region of the mouse GABRA4 gene that encodes the alpha4 subunit and found that the promoter has multiple transcriptional initiation sites and lacks a TATA box. The minimal promoter for GABRA4 spans the region between -444 to -19 bp relative to the coding ATG and shows high activity in cultured mouse cortical neurons. Both Sp3 and Sp4 transcription factors can interact with the two Sp1 binding sites within the minimal promoter and are critical for maximal activity of the promoter in neurons.


Current status and recent advantages in derivatization procedures in human doping control.
Derivatization is one of the most important steps during sample preparation in doping control analysis. Its main purpose is the enhancement of chromatographic separation and mass spectrometric detection of analytes in the full range of laboratory doping control activities. Its application is shown to broaden the detectable range of compounds, even in LC-MS analysis, where derivatization is not a prerequisite. The impact of derivatization initiates from the stage of the metabolic studies of doping agents up to the discovery of doping markers, by inclusion of the screening and confirmation procedures of prohibited substances in athlete's urine samples. Derivatization renders an unlimited number of opportunities to advanced analyte detection.


A Two-Step Synthesis of 2-Spiropiperidines.
A general two-step synthesis of 2-spiropiperidines has been developed. δ-Amino-β-ketoesters can be reacted with cyclic ketones to generate 2-spiropiperidines in good to excellent yields. The 2-spiropiperidines formed occupy an under-explored region of 3D-chemical space and are novel scaffolds for use in drug discovery programs. These 2-spiropiperidines can be further functionalised to generate small highly sp3 -rich structures, which exhibit an excellent likeness to lead-molecules in drug discovery.


Optical amplification from high vibrational states of ionized nitrogen molecules generated by 800-nm femtosecond laser pulses.
We experimentally investigated the interaction between nitrogen molecules and intense femtosecond laser pulses. When irradiated by an 800-nm pump laser and a delayed 355-nm seed laser, the spectral lines around 353.3 nm and 353.8 nm are observed to be greatly amplified, no matter whether the pump laser is circularly or linearly polarized. The two spectral lines correspond to the transition of N2+ (B, ν' = 5 → X, ν = 4) and N2+ (B, ν' = 4 → X, ν = 3), respectively. In comparison with the spectral lines related with ground vibrational states of nitrogen molecular ion, the observed amplification exhibits different polarization dependence of the pump laser. This distinctive change can be explained by the population variation of high vibrational states caused by the pump laser with different polarizations.


Preclinical pharmacokinetics, interspecies scaling, and pharmacokinetics of a Phase I clinical trial of TTAC-0001, a fully human monoclonal antibody against vascular endothelial growth factor 2.
VEGF is a highly selective mitogen that serves as the central regulator of tumor angiogenesis by mediating endothelial proliferation, permeability, and survival. Tanibirumab (TTAC-0001) is a fully human IgG1 monoclonal antibody derived from a fully human naïve single-chain variable fragment (ScFv) phage library that was developed to inhibit the effects of VEGF in the treatment of solid tumors, especially those of the brain. In the present study, we conducted intravenous pharmacokinetic studies of TTAC-0001 in mice, rats, and cynomolgus monkeys. At the doses studied (3 mg/kg, 10 mg/kg, 30 mg/kg), TTAC-0001 exhibited dose proportionality in mice and monkeys. At a dose of ~10 mg/kg, the clearance of TTAC-0001 from serum was 0.017 mL/h in mice, 0.35 mL/h in rats, and 2.19 mL/h in cynomolgus monkeys, and the terminal half-life ranged from 20-30 h among the three species. Pharmacokinetic data in mice, rats, and cynomolgus monkeys were used to predict the pharmacokinetics of TTAC-0001 in humans using allometric scaling. The predicted serum clearance of TTAC-0001 in humans was 102.45 mL/h and the terminal half-life was 27.52 h. The maximum life span-corrected clearance value was 72.92 mL/h. The observed clearance in humans was more similar to the predicted scaled clearance. We investigated the pharmacokinetics of TTAC-0001 in mice, rats, and cynomolgus monkeys after intravenous administration. At the doses studied, TTAC-0001 exhibited dose proportionality in mice and monkeys. The scaled pharmacokinetics of TTAC-0001 reported here was useful for designing first-in-human studies. Allometric scaling in the therapeutic antibody is feasible.


UK survey demonstrates a wide range of impacts attributable to clinical librarian services.
To understand the impact of the UK Clinical Librarian (CL) workforce and benchmark the results against a study undertaken in the North West region of the English National Health Service (NHS). An online survey was distributed by CLs to their service users regarding literature searches that had been carried out on their behalf in the 6 months from April to October 2017. Interviews were later carried out in person with selected respondents to the questionnaires. CLs across the UK contribute to a wide range of outcomes, with 41% of search requests contributing to the choice of intervention, and 41% also to the advice offered by the clinician requester to a patient or their carer. These results are in line with the previous work undertaken in the North West. CLs provide diverse services to clinical teams. They support the continuing professional development and personal research needs of team members, service development needs of organisations, and the information provided contributes to improved quality and safety of patient care. The survey confirms the findings of the earlier NW study. It demonstrates the impact of services based around literature searching on patient care.


[Reexpansion Pulmonary Edema during Emergency Surgery in a Patient with Spontaneous Hemopneumothorax].
A 43-year-old male patient with spontaneous hemopneumothorax of the right lung underwent emergency video-assisted thoracoscopic surgery for drainage, hemostasis and bullae resection. Fifteen minutes after reexpansion of the right lung, we found bubbly sputum coming out from the right tracheal tube and cloudy shadow in the right field of his chest X-ray. The occurrence of reexpansion pulmonary edema (RPE) was considered. Subsequent mechanical ventilation with PEEP and administration of steroid and diuretic was done as his treatment. His respiratory state was stabiized in the next two days. As the lung collapse following spontaneous hemopneumothorax often becomes more severe, we should pay attention to the occurrence of RPE after expansion of affected side lung. And, if it occurred, appropriate and prompt treatment as above should be done because of its high mortality.


VEGF is involved in the increase of dermal microvascular permeability induced by tryptase.
Tryptases are predominantly mast cell-specific serine proteases with pleiotropic biological activities and play a critical role in skin allergic reactions, which are manifested with rapid edema and increases of vascular permeability. The exact mechanisms of mast cell tryptase promoting vascular permeability, however, are unclear and, therefore, we investigated the effect and mechanism of tryptase or human mast cells (HMC-1) supernatant on the permeability of human dermal microvascular endothelial cells (HDMECs). Both tryptase and HMC-1 supernatant increased permeability of HDMECs significantly, which was resisted by tryptase inhibitor APC366 and partially reversed by anti-VEGF antibody and SU5614 (catalytic inhibitor of VEGFR). Furthermore, addition of tryptase to HDMECs caused a significant increase of mRNA and protein levels of VEGF and its receptors (Flt-1 and Flk-1) by Real-time RT-PCR and Western blot, respectively. These results strongly suggest an important role of VEGF on the permeability enhancement induced by tryptase, which may lead to novel means of controlling allergic reaction in skin.


Testosterone replacement in men with andropause: an overview.
Serum testosterone levels decline progressively as men age, with resulting pathophysiological changes. Because the onset of andropause is gradual and many of its symptoms mirror those associated with medications or disease states common in the elderly, a clinical diagnosis can be difficult to make. Additionally, because of a lack of established normal testosterone levels for different age groups, as well as confusion regarding what subset of testosterone to measure, simply testing testosterone levels is inadequate. Although clinical studies have shown testosterone supplementation to be safe, no long-term placebo-controlled trials have been performed. Among the possible side effects of testosterone replacement therapy is an increased risk of prostate cancer. While there is no evidence that supplemental testosterone will initiate prostate cancer or cause clinically significant progression of an established occult malignancy, the initial evaluation of patients prior to administration of testosterone requires screening for prostatic carcinoma. A number of formulations of testosterone are currently available; transdermal gels, although expensive, are the preferred modality because of their ease of administration.


Nursing interventions and patient outcomes: a meta-analysis of studies.
The 84 subject-studies and 4,146 individual subjects in this meta-analysis were obtained from nurse-conducted experimental research over an 8-year period. The entire universe of accessible subject-studies that met criteria was included. Although both published and unpublished research were included to protect the study from publication bias, there was no statistically significant difference in findings. The mean effect size for the sample of comparisons from the 84 studies was .59. The associated U3 value of 72.2 and r of .28 indicate that patients who receive research-based nursing interventions can expect 28% better outcomes than 72% of the patients who receive standard nursing care.


Antibody to myelin basic protein in extracts of multiple sclerosis brain.
Autoimmunity to a neural antigen is a suspected cause of multiple sclerosis (MS), and a candidate autoantigen is myelin basic protein (MBP). Accordingly, saline extracts of brain from patients with MS and other diseases were prepared and the content of immunoglobulin (Ig) determined. Antibody to MBP was measured with a highly-sensitive solid-phase radioimmunoassay using 125I-staphylococcal Protein A. Anti-MBP activity was detected in brain extracts of all eleven MS patients, and in seven out of the eight brain extracts from the patients with other diseases; however the level of anti-MBP activity was significantly higher in the MS extracts (P less than 0.01). Analysis of the MS brain extracts after purification by affinity chromatography columns revealed that the anti-MBP activity was specifically mediated by IgG and resided in the IgG1, IgG2, and/or IgG4 subclasses.


Intracellular location of thymidylate synthase and its state of phosphorylation.
Thymidylate synthase (TS), an enzyme that is essential for DNA synthesis, was found to be associated mainly with the nucleolar region of H35 rat hepatoma cells, as determined both by immunogold electron microscopy and by autoradiography. In the latter case, the location of TS was established through the use of [6-3H]5-fluorodeoxyuridine, which forms a tight ternary complex of TS with 5-fluorodeoxyuridylate (FdUMP) and 5, 10-methylenetetrahydrofolylpolyglutamate within the cell. However, with H35 cells containing 50-100-fold greater amounts of TS than unmodified H35 cells, the enzyme, although still in the nucleus, was located primarily in the cytoplasm as shown by autoradiography and immunohistochemistry. In addition, TS was also present in mitochondrial extracts of both cell lines, as determined by enzyme activity measurements and by ternary complex formation with [32P]FdUMP and 5,10-methylenetetrahydrofolate. Another unique observation is that the enzyme appears to be a phosphoprotein, similar to that found for other proteins associated with cell division and signal transduction. The significance of these findings relative to the role of TS in cell division remains to be determined, but suggest that this enzyme's contribution to the cell cycle may be more complex than believed previously.


TRPA1 contributed to the neuropathic pain induced by docetaxel treatment.
Peripheral mechanical neuropathic pain is a serious side effect of docetaxel chemotherapy for cancer. However, the underlying mechanism for this side effect is unknown. In the present study, we found that docetaxel treatment induced mechanical allodynia in rats. We further revealed that the transient receptor potential ankyrin subtype 1 protein (TRPA1) protein level is upregulated and the TRPA1 activator allyl isothiocyanate induced larger ion currents in the dorsal root ganglion neurons from the docetaxel treated rats. In addition, application the TRPA1 blocker Ap18 reversed the docetaxel-induced mechanical hypersensitivity. We suggest that the docetaxel-induced mechanical allodynia is mediated by upregulation of TRPA1 in dorsal root ganglion neurons.