Secondary-care costs associated with lung cancer diagnosed at emergency hospitalisation in the United Kingdom.
Lung cancer diagnosis during emergency hospital admission has been associated with higher early secondary-care costs and lower longer-term costs than outpatient diagnoses. This retrospective cohort study analyses the secondary-care costs of 3274 consecutive patients with lung cancer. Patients diagnosed during emergency admissions incurred greater costs during the first month and had a worse prognosis compared with outpatient diagnoses. In patients who remained alive, costs after the first month were comparable between diagnostic routes. In addition to improving patient experience and outcome, strategies to increase earlier diagnosis may reduce the additional healthcare costs associated with this route to diagnosis.


Comparison of SARS-CoV-2 PCR-Based Detection Using Saliva or Nasopharyngeal Swab Specimens in Asymptomatic Populations.
The coronavirus disease 2019 (COVID-19) pandemic has challenged clinical diagnostic operations due to supply shortages and high staffing needs to collect nasopharyngeal (NP) swab samples. Saliva is an easily accessible alternative specimen type to overcome some of these challenges. In this study, we first used paired saliva and NP swab specimens (n = 128) to compare test performance characteristics with three RNA extraction platforms, i.e., Maxwell RSC (Promega), MagNA Pure 96 (Roche), and KingFisher Flex (Thermo Fisher Scientific), together with two PCR chemistries, i.e., severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) (2019-nCoV) Centers for Disease Control and Prevention (CDC) quantitative PCR (qPCR) probe assay (Integrated DNA Technologies) and TagPath COVID-19 combination kit (Thermo Fisher Scientific). This study demonstrated that both saliva and NP swab specimens performed well, with 97% agreement when tested by the CDC qPCR chemistry using Maxwell and MagNA Pure RNA extraction platforms. We then compared 12 weeks of saliva and NP swab testing results using two independent asymptomatic populations, including a community surveillance program using saliva samples only (n = 466) and preoperative screening using NP swab samples only (n = 8,461). The positive detection rates among participants with either saliva or NP swab samples were 1.07% and 1.12%, respectively, which confirms the low pretest probabilities for COVID-19 infections in asymptomatic populations. Notably, there was no increased proportion of low-titer cases (inconclusive results) reported in the asymptomatic groups, compared with the all-comers groups (0.21% and 0.66%, respectively, in the community population and 0.25% and 0.49%, respectively, in the preoperative population); this suggests that low-viral-titer carriers can be found similarly in both groups with saliva or NP swab specimens. In summary, saliva can be considered a good alternative for noninvasive but well-instructed self-collection. IMPORTANCE Our study shows that saliva is a noninvasive respiratory secretion sample type that contains equal or more host materials (RNase P), compared with those contained in the corresponding NP swab specimens, in 103 paired samples. SARS-CoV-2 detection with two RNA extraction platforms, Maxwell and MagNA Pure, with CDC qPCR chemistry showed similar test sensitivities for paired specimens. We then analyzed SARS-CoV-2 detections rates in two independent groups of asymptomatic participants, i.e., a group at a community screening station with supervised saliva collection only (n = 466) and a preoperative screening group (n = 8,461) with NP swabbing only. Similar detection rates of 1.07% for the community group and 1.12% for the preoperative group supported the similar test performances in these groups predicted to have low pretest probabilities of infection. With mindful preparation, saliva can be considered for schools and clinical participants when adequate collection education can be provided and compliance can be established.


Suicidality and Its Risk Factors in Korean People with Epilepsy: A MEPSY Study.
People with epilepsy (PWE) are more likely to experience suicidality, with suicidal ideation and attempts, than people without epilepsy (PWoE). The aims of the present study were to determine 1) the characteristics of suicidality in Korean PWE, 2) whether PWE with suicidality receive psychiatric intervention, and 3) the risk factors for suicidality. Patients who consecutively visited epilepsy clinics at secondary- and tertiary-care hospitals were recruited (n=684), along with age- and sex-matched PWoE (n=229). The presence of current major depressive disorder (MDD), generalized anxiety disorder (GAD), and/or suicidality was established using the Mini International Neuropsychiatric Interview-Plus Version 5.0.0. The Korean version of the Liverpool Adverse Events Profile (K-LAEP) was applied to detect adverse effects of antiepileptic drugs (AEDs). Suicidality was present in 208 (30.4%) of the 684 PWE. The rate of suicidality was 4.6 times higher among PWE than PWoE, and 108 (15.7%) PWE had suicidal ideation and had attempted suicide. Among those who had attempted suicide, 40.7% had made at least two attempts. The most common method of suicide attempt was drug overdose (34.9%). Unfortunately, of the 208 PWE with suicidality, 136 (65.4%) did not receive psychiatric intervention. The risk factors for suicidality were MDD [odds ratio (OR)=6.448, 95% confidence interval (CI)=3.739-11.120, p<0.001], GAD (OR=3.561, 95% CI=1.966-6.452, p<0.001), item scores of 3 or 4 on the K-LAEP (OR=2.688, 95% CI=1.647-4.387, p<0.001), and a history of febrile convulsion (OR= 2.188, 95% CI=1.318-3.632, p=0.002). Suicidality is more prevalent in PWE than in PWoE. Clinicians should monitor psychiatric disorders and the adverse effects of AEDs in PWE in an attempt to reduce the incidence of suicidal ideation or suicide attempts in this patient population.


Foliar response of black cherry (Prunus serotina) clones to ambient ozone exposure in central Pennsylvania.
During late summer of 1996 and 1997 we examined ozone-induced foliar injury in a plantation of 111 black cherry trees (ramets) comprising 15 clones originating from wild ortets growing in the Allegheny National Forest, Pennsylvania, and the Monongahela National Forest, West Virginia. The experimental plantation was a clonal seed orchard in Centre County, Pennsylvania, started in 1971 using ortet buds grafted onto seedling rootstocks of mixed origin. Clones differed significantly in severity of foliar injury symptoms (F=31.83, p<0.001). One clone (R-12) had significantly more foliar injury with >50% leaf area affected than other clones during both years. In contrast, clone R-14, which is from the same area in northcentral Pennsylvania as R-12, exhibited significantly less injury (LAA<6%). Although ambient O(3) concentrations were similar in both years, foliar injury was significantly greater (15.7%) in 1996 than in 1997 (9.9%). This is probably explained by lower stomatal conductance in 1997 caused by drier and hotter weather patterns in June and July of that year. Despite very different weather patterns and overall levels of injury in 1996 and 1997, mean clonal injury was significantly correlated between both years of assessment (r=0.92, p<0.001). Within tree crowns, foliage in lower and inner crown positions was significantly more injured than foliage in upper and exterior crown positions. There was no evidence of geographically based population differences in sensitivity to foliar O(3) injury. On the contrary, results demonstrate that wild genotypes of proximal geographic origin may differ greatly in sensitivity.


Albuminuria as a risk factor for peripheral arterial disease in a general population: the Hisayama study.
To elucidate the relationship between albuminuria and the prevalence of peripheral arterial disease (PAD), and to examine the effect of albuminuria on the ability to assess the likelihood of PAD in a general Japanese population. In 3,061 community-dwelling subjects aged. 40 years, we investigated the association of urinary albumin-creatinine ratio (UACR) levels with the prevalence of PAD, defined as an ankle-brachial index < 0.9. The odds ratio for the presence of PAD was estimated using the logistic regression model. To compare the accuracy of the assessment for the likelihood of prevalent PAD between models adjusted for potential risk factors with and without UACR levels, the receiver operating characteristic (ROC) curves were plotted. Overall, 1.47% of the study participants had PAD. The age- and sex-adjusted prevalence of PAD increased linearly for UACR levels of < 5.6, 5.6-10.8, 10.9-29.9, 30.0-300.0, and >300.0 mg/g, being 0.34, 0.80, 2.02, 2.50, and 2.53%, respectively (p for trend <0.001). The multivariate-adjusted odds ratio for the presence of PAD was 1.85 (95% confidence interval 1.12-3.06) for every 10-fold increment in UACR. The area under the ROC curve significantly increased when UACR levels were incorporated into a model with potential risk factors for PAD (0.80 vs. 0.77, p= 0.02). Greater UACR levels are associated linearly with a higher prevalence of PAD, even within the normoalbuminuric range, in the general Japanese population, and combining UACR levels with potential risk factors substantially improves the performance to assess the likelihood of PAD.


Addition of a carboxy-terminal tail to the normally tailless gonadotropin-releasing hormone receptor impairs fertility in female mice.
Gonadotropin-releasing hormone (GnRH) is the primary neuropeptide controlling reproduction in vertebrates. GnRH stimulates follicle-stimulating hormone (FSH) and luteinizing hormone (LH) synthesis via a G-protein-coupled receptor, GnRHR, in the pituitary gland. In mammals, GnRHR lacks a C-terminal cytosolic tail (Ctail) and does not exhibit homologous desensitization. This might be an evolutionary adaptation that enables LH surge generation and ovulation. To test this idea, we fused the chicken GnRHR Ctail to the endogenous murine GnRHR in a transgenic model. The LH surge was blunted, but not blocked in these mice. In contrast, they showed reductions in FSH production, ovarian follicle development, and fertility. Addition of the Ctail altered the nature of agonist-induced calcium signaling required for normal FSH production. The loss of the GnRHR Ctail during mammalian evolution is unlikely to have conferred a selective advantage by enabling the LH surge. The adaptive significance of this specialization remains to be determined.


Refractive Outcomes After Trabecular Micro-Bypass Stents (iStent Inject) with Cataract Extraction in Open-Angle Glaucoma.
Simultaneous cataract and glaucoma surgery has traditionally been challenging for the anterior segment surgeon. The introduction of minimally invasive glaucoma surgery (MIGS) in conjunction with cataract surgery appears safe and effective in lowering intraocular pressure. Although a significant visual impact leading from the combined procedure is unexpected, we aim to describe the refractive outcomes in a cohort of patients undergoing simultaneous cataract removal and iStent inject and discuss the potential implications of combined surgery in patients with co-existent glaucoma. This is a retrospective consecutive case series inclusive of patients undergoing combined femtosecond laser-assisted cataract surgery and the insertion of two trabecular micro-bypass stents (iStent inject). Visual acuity, refraction and astigmatic vector analysis were collated and analysed from the preoperative and 4 weeks postoperative visits. One hundred and six eyes of 89 patients from 2 surgeons were included in the original cohort. The mean absolute difference from target refraction was 0.36 ± 0.25D. 73.9% of eyes were within ± 0.5D of the refractive target and 98.9% of eyes were within ± 1.00D. 73.8% of eyes had 0.5D or less residual refractive astigmatism following the procedure. We present a novel cohort of glaucoma patients undergoing combined trabecular micro-bypass stents (iStent inject) and cataract surgery achieving excellent refractive outcomes. The results of this study indicate that this second-generation device is refractively neutral.


Protocol for preparation of LiCl-based ultra-hygroscopic curdlan heat exchanger for dehumidification.
Desiccant-coated heat exchangers provide a practical solution for the efficient removal of moisture from the air. Here, we present a protocol to synthesize an ultra-hygroscopic polymer to develop a LiCl loaded in curdlan hydrogel (LiCl@Cur)-coated heat exchanger for deep dehumidification. We describe steps for preparing the curdlan gel solution, hydrogel, LiCl solution, and LiCl@Cur. We then detail procedures for preparing curdlan-coated and LiCl@Cur-coated heat exchangers. The coated heat exchanger described in this protocol has a maximum dehumidification capacity of 12 g/kg. For complete details on the use and execution of this protocol, please refer to Pan et al. (2023).1.


Marangoni-induced reversal of meniscus-climbing microdroplets.
Small water droplets or particles located at an oil meniscus typically climb the meniscus due to unbalanced capillary forces. Here, we introduce a size-dependent reversal of this meniscus-climbing behavior, where upon cooling of the underlying substrate, droplets of different sizes concurrently ascend and descend the meniscus. We show that microscopic Marangoni convection cells within the oil meniscus are responsible for this phenomenon. While dynamics of relatively larger water microdroplets are still dominated by unbalanced capillary forces and hence ascend the meniscus, smaller droplets are carried by the surface flow and consequently descend the meniscus. We further demonstrate that the magnitude and direction of the convection cells depend on the meniscus geometry and the substrate temperature and introduce a modified Marangoni number that well predicts their strength. Our findings provide a new approach to manipulating droplets on a liquid meniscus that could have applications in material self-assembly, biological sensing and testing, or phase change heat transfer.


Addition of [18 F]Fluorodeoxyglucose Positron Emission Tomography With Computed Tomography to Cross-Sectional Imaging Improves Staging and Alters Management in Hepatocellular Carcinoma.
In this work, we characterize the value of positron emission tomography (PET) with computed tomography (CT) in combination with cross-sectional imaging for staging and prognostication of hepatocellular carcinoma (HCC) patients. In this retrospective cohort study, HCC patients underwent PET-CT after initial staging with contrast-enhanced CT or magnetic resonance imaging (MRI). The benefit of PET-CT was measured by the identification of new HCC lesions, and potential harm was quantified by the number of false positives and subsequent diagnostic evaluation. We used multivariate Cox regression analysis to evaluate the association between the highest grade on PET-CT with the risk of extrahepatic metastasis, progression-free, and overall survival. Among 148 patients, PET-CT detected additional extrahepatic metastasis in 11.9% of treatment-naïve and 13.8% of treatment-experienced patients. PET-CT changed the Barcelona Clinic Liver Cancer (BCLC) staging in 5.9% of treatment-naïve and 18.8% of treatment-experienced patients compared with CT/MRI alone, changing HCC management in 9.9% and 21.3% of patients, respectively. Of the patients, 5% (n = 8) experienced severe physical harm requiring additional procedures to evaluate extrahepatic findings. High tumor grade on PET-CT was independently associated with a higher likelihood of extrahepatic metastasis (hazard ratio [HR], 17.1; 95% confidence interval [CI], 3.6-81.5) and worse overall survival (HR, 2.4; 95% CI, 1.4-4.3). Treatment-experienced patients (versus treatment-naïve patients; HR, 9.7; 95% CI, 1.9-49.4) and BCLC stage A (HR, 8.2; 95% CI, 1.5-45.9; P < 0.01) and BCLC stage B (HR, 20.6; 95% CI, 1.5-282.2; P < 0.05) were more likely to have an upstaging with PET-CT compared with BCLC stage C (reference). PET-CT provides prognostic information and improves tumor staging beyond CT/MRI alone, with subsequent changes in management for patients with HCC.


Proteome profiling of immortalization-to-senescence transition of human breast epithelial cells identified MAP2K3 as a senescence-promoting protein which is downregulated in human breast cancer.
immortalization is one of the first changes in cells undergoing carcinogenic transformation. Proteome profiling of the immortalization-senescence transition is expected to provide insights into the molecular mechanisms of early tumorigenesis. 2-DE and MALDI-MS were used to identify proteins in primary human breast epithelial cells, relevant to the immortalization-senescence transition. Cell and molecular biology and immunohistochemistry were used to validate involvement of mitogen-activated protein kinase kinase 3 (MAP2K3) in the immortalization-senescence transition. we identified 71 proteins whose expression changed upon induction of senescence. The identified proteins include regulators of cell growth, death, cell assembly and organization. Analysis of the network formed by the identified proteins suggested that the immortalization-to-senescence transition could affect regulators of the cell cycle, protein synthesis, transport, post-translational modifications, DNA recombination and repair, and lipid and amino acid metabolism. We observed that MAP2K3 was downregulated in immortal human breast epithelial cells and that upregulation of MAP2K3 expression promoted cell senescence. Decreased expression of MAP2K3 was observed in human breast infiltrating ductal carcinomas, as compared to non-cancerous human breast tissues. we described a proteome profile of the immortalization-to-senescence transition for human breast epithelial cells, and identified MAP2K3 as a protein that promotes senescence in these cells.


The direct arylation of allylic sp(3) C-H bonds via organic and photoredox catalysis.
The direct functionalization of unactivated sp(3) C-H bonds is still one of the most challenging problems facing synthetic organic chemists. The appeal of such transformations derives from their capacity to facilitate the construction of complex organic molecules via the coupling of simple and otherwise inert building blocks, without introducing extraneous functional groups. Despite notable recent efforts, the establishment of general and mild strategies for the engagement of sp(3) C-H bonds in C-C bond forming reactions has proved difficult. Within this context, the discovery of chemical transformations that are able to directly functionalize allylic methyl, methylene and methine carbons in a catalytic manner is a priority. Although protocols for direct oxidation and amination of allylic C-H bonds (that is, C-H bonds where an adjacent carbon is involved in a C = C bond) have become widely established, the engagement of allylic substrates in C-C bond forming reactions has thus far required the use of pre-functionalized coupling partners. In particular, the direct arylation of non-functionalized allylic systems would enable access to a series of known pharmacophores (molecular features responsible for a drug's action), though a general solution to this long-standing challenge remains elusive. Here we report the use of both photoredox and organic catalysis to accomplish a mild, broadly effective direct allylic C-H arylation. This C-C bond forming reaction readily accommodates a broad range of alkene and electron-deficient arene reactants, and has been used in the direct arylation of benzylic C-H bonds.


The experience of a good day: a phenomenological study to explain a good day as experienced by a newly qualified RN.
The main aim of this study was to provide an explanation of the newly qualified nurse's experience and description of a good day. Secondly it sought to provide an explanation of how a good day made them feel about nursing. By identifying the main components of a good day and what positively effects the experience of a working day for a newly qualified nurse, it may be possible to move towards an increase in the occurrence of the components and emulation of a good day. While there is evidence to suggest that positive experiences within nursing increase job satisfaction and aid retention to the profession, the experience of a good day and what constitutes a good day for a newly qualified nurse has not been explored. The main components of a good day have not been identified and no work has been carried out to ascertain how these days make nurses feel about their chosen career. The aims of the study lent themselves to a phenomenological descriptive approach to research, the objective of which is identification of the essence of behaviour. Eight newly qualified nurses agreed to take part in the research. Each participant was interviewed twice, and in addition one group interview was arranged to clarify themes. The interviews, which were audio taped, were informal allowing the participants to answer in an open and unstructured manner. Once completed, all the tapes were transcribed and immersion and analysis of the data led to 5 themes naturally emerging as the components of a good day. The identified themes were, doing something well, good relationships with patients, feeling that you've achieved something, getting the work done and you need team work. In addition, although not a theme of a good day but of great importance was the description of 'that wonderful feeling at the end of a good day'. These themes contributed to feelings of job satisfaction and the pleasure of nursing. More specifically the concept of knowing the patient both from a personal level and knowing about their care and condition has been identified as a main component of a good day. This knowing the patient had a knock on effect in that when this was part of the day it was easier to prioritise and get the work done. These nurses were striving to develop a level of care and competency and, unlike expert nurses, did not have vast experience and knowledge to underpin their actions. They need to ensure that all levels of knowledge are addressed before they can carry out care. Additionally the team dynamics and the way the team work together had a significant effect on the day.


Using Omaha System Documentation to Understand Physical Activity among Rural Women.
Examine factors related to physical activity among rural women using Omaha System data. Mixed methods: quantitative analysis of de-identified clinical data and thematic analysis of public health nurse (PHN) focus group data. Rural women who received PHN services (N = 852) and purposively selected PHNs (N = 12). Omaha System problems; signs/symptoms; knowledge, behavior, and status ratings; demographics. Physical activity behavior among rural women was inconsistently appropriate (M = 3.27). Age, body mass index, physiological, psychosocial, and environmental domain problems, and physical activity knowledge explained 33.2% of physical activity behavior variance. Almost all women for whom specific problems were documented had insufficient physical activity. This included physiological problems of cognition and pregnancy; psychosocial problems of abuse, caretaking/parenting, mental health, and interpersonal relationship; and environmental problems of neighborhood/workplace safety and income. Having problems in two or more domains was significantly associated with insufficient physical activity behavior (p < .001). PHNs validated findings and expressed concerns that client complexity, priorities, and time constraints may affect client assessment and documentation. PHNs should assess rural women who are not physically active for potentially serious physiological, psychosocial, and environmental problems. Departmental policies requiring assessment of Omaha System data across programs enabled population health measurement and research.


Electromechanical coupling between skeletal and cardiac muscle. Implications for infarct repair.
Skeletal myoblasts form grafts of mature muscle in injured hearts, and these grafts contract when exogenously stimulated. It is not known, however, whether cardiac muscle can form electromechanical junctions with skeletal muscle and induce its synchronous contraction. Here, we report that undifferentiated rat skeletal myoblasts expressed N-cadherin and connexin43, major adhesion and gap junction proteins of the intercalated disk, yet both proteins were markedly downregulated after differentiation into myo-tubes. Similarly, differentiated skeletal muscle grafts in injured hearts had no detectable N-cadherin or connexin43; hence, electromechanical coupling did not occur after in vivo grafting. In contrast, when neonatal or adult cardiomyocytes were cocultured with skeletal muscle, approximately 10% of the skeletal myotubes contracted in synchrony with adjacent cardiomyocytes. Isoproterenol increased myotube contraction rates by 25% in coculture without affecting myotubes in monoculture, indicating the cardiomyocytes were the pacemakers. The gap junction inhibitor heptanol aborted myotube contractions but left spontaneous contractions of individual cardiomyocytes intact, suggesting myotubes were activated via gap junctions. Confocal microscopy revealed the expression of cadherin and connexin43 at junctions between myotubes and neonatal or adult cardiomyocytes in vitro. After microinjection, myotubes transferred dye to neonatal cardiomyocytes via gap junctions. Calcium imaging revealed synchronous calcium transients in cardiomyocytes and myotubes. Thus, cardiomyocytes can form electromechanical junctions with some skeletal myotubes in coculture and induce their synchronous contraction via gap junctions. Although the mechanism remains to be determined, if similar junctions could be induced in vivo, they might be sufficient to make skeletal muscle grafts beat synchronously with host myocardium.


Efficacy and safety of human adipose tissue-derived mesenchymal stem cells for supporting hematopoiesis.
We have demonstrated that adipose tissue-derived mesenchymal stem cells (ADSCs) from mice are capable of reconstituting the hematopoietic microenvironment, and facilitate hematopoiesis more effectively than bone marrow-derived mesenchymal stem cells (BMSCs) in mouse. The ready accessibility of fat tissue rich in MSCs and the higher hematopoiesis-supporting capacities of ADSCs suggest that ADSCs might represent a new therapeutic modality for the regeneration of impaired hematopoiesis. As a further step towards their use in clinical practice, we established human BMSCs and ADSCs from healthy volunteers of similar age, and compared their proliferation capacities, hematopoiesis-supporting properties, and safety. In vitro cell proliferation studies revealed that ADSCs have a higher population doubling number than BMSCs. In vitro co-culture assays showed that ADSCs not only support human CD34(+) peripheral blood stem cells (PBSCs), but also yield significantly more non-adherent hematic cells than BMSCs. In vitro progenitor assays revealed that ADSCs promote a higher frequency of early progenitors than do BMSCs. Interestingly, BM cellularity in irradiated mice that had received ADSCs tended to be higher than that of mice treated with BMSCs. When MSCs were injected into the BM cavity of tibiae, we observed no evidence of MSC-induced toxicity either during or after treatment. In addition, no microscopic abnormalities were observed in the bone marrow and major organs.


Ultrasound-based renal sympathetic denervation for the treatment of therapy-resistant hypertension: a single-center experience.
The effectiveness of renal sympathetic denervation (RDN) as a treatment for therapy-resistant hypertension has been doubted as the Simplicity-HTN-3 trial was unable to show any treatment benefit over sham procedure. This might partly be explained by a high procedural variability in treatment with radiofrequency-based catheters. Recently, newer systems for RDN, like ultrasound-based devices, have been introduced into practice. To date however, data on their effectiveness for the treatment of resistant hypertension are scarce. We sought to evaluate the safety and effectiveness of an ultrasound-based, balloon-irrigated RDN catheter in a larger single-center cohort. Patients with therapy-resistant hypertension [average blood pressure (BP) >135 mmHg SBP or >90 mmHg DBP in ambulatory BP measurement despite at least three antihypertensive drugs, including at least one diuretic] underwent ultrasound-based RDN. Treatment effect was assessed by comparing BP values at baseline and 3 months after the procedure. Patients underwent renal artery duplex sonography or MRI before and after RDN to exclude renal artery stenosis. Fifty consecutive patients underwent ultrasound-based RDN, of which 25 had undergone an unsuccessful radiofrequency RDN before. Mean SBP change at 3 months was -9.7 ± 12.6/-10.6 ± 13.7/-8.2 ± 15.2 mmHg (ambulatory 24-h mean/daytime/night, P < 0.001 for all) and DBP changed by -5.1 ± 7.4/-5.8 ± 7.8/-3.9 ± 10.3 mmHg (P ≤ 0.001/<0.001/0.01). No new renal artery stenosis could be detected after RDN. Ultrasound-based RDN seems to be well tolerated and effective for the treatment of patients with therapy-resistant hypertension.


Plotting the course to remission: the search for better outcomes in the treatment of depression.
Depression includes a wide range of symptoms that can impair a person's psychosocial and physical functioning. This impairment can lead to decreased productivity, increased health care utilization, alcohol and substance abuse, and an increased risk of suicide. While the treatment of depression has significantly advanced over the past 30 years, there is still room for improvement. Full remission of depressive symptoms is often elusive, and many patients never achieve full relief from their depression despite being regarded as responders to antidepressant treatment. Current treatments for depression tend to focus on emotional symptoms, not the physical and anxious symptoms also associated with depression. However, the physical and anxious symptoms of depression can be serious and sometimes more prominent than the emotional symptoms of depression, especially among special populations such as women. New treatment strategies, such as dual-acting agents and the combination of pharmacotherapy and psychotherapy, target the emotional and anxious symptoms of depression as well as symptoms associated with pain. In order to increase response and remission, depression should be seen as an illness comprising not only emotional symptoms but physical and anxious symptoms as well.


Stagonospora nodorum: cause of stagonospora nodorum blotch of wheat.
SUMMARY Stagonospora nodorum is an important pathogen of wheat and related cereals, causing both a leaf and glume blotch. This review summarizes recent advances in our understanding of taxonomy, control and pathogenicity of this species. Stagonospora (syn. Septoria) nodorum (Berk.) Castell. and Germano [teleomorph: Phaeosphaeria (syn. Leptosphaeria) nodorum (Müll.) Hedjar.], kingdom Fungi, phylum Ascomycota, subphylum Euascomycota, class Dothideomycetes, order Pleosporales, family Phaeosphaeriaceae, genus Phaeosphaeria, species nodorum. Wheat, Triticum aestivum, T. durum, Triticale, are the main hosts but other cereals and wild grasses have been reported to harbour S. nodorum. Disease symptoms are lens-shaped necrotic lesions on leaves, girdling necrosis on stems (especially the nodes, hence 'nodorum') and lesions on glumes. Mature lesions produce pycnidia scattered throughout the lesions, especially as tissue senesces. http://ocid.nacse.org/research/deephyphae/htmls/asco_taxlist_spat.html (taxonomic information), http://ohioline.osu.edu/ac-fact/0002.html (disease information), http://wwwacnfp.murdoch.edu.au/ (ACNFP homepage), http://www.broad.mit.edu/annotation/fungi/stagonospora_nodorum/index.html (genome sequence homepage), http://cogeme.ex.ac.uk/efungi/ (genome sequence annotation and analysis).


Integrative monitoring of marine and freshwater harmful algae in Washington State for public health protection.
The more frequent occurrence of both marine and freshwater toxic algal blooms and recent problems with new toxic events have increased the risk for illness and negatively impacted sustainable public access to safe shellfish and recreational waters in Washington State. Marine toxins that affect safe shellfish harvest in the state are the saxitoxins that cause paralytic shellfish poisoning (PSP), domoic acid that causes amnesic shellfish poisoning (ASP) and the first ever US closure in 2011 due to diarrhetic shellfish toxins that cause diarrhetic shellfish poisoning (DSP). Likewise, the freshwater toxins microcystins, anatoxin-a, cylindrospermopsins, and saxitoxins have been measured in state lakes, although cylindrospermopsins have not yet been measured above state regulatory guidance levels. This increased incidence of harmful algal blooms (HABs) has necessitated the partnering of state regulatory programs with citizen and user-fee sponsored monitoring efforts such as SoundToxins, the Olympic Region Harmful Algal Bloom (ORHAB) partnership and the state's freshwater harmful algal bloom passive (opportunistic) surveillance program that allow citizens to share their observations with scientists. Through such integrated programs that provide an effective interface between formalized state and federal programs and observations by the general public, county staff and trained citizen volunteers, the best possible early warning systems can be instituted for surveillance of known HABs, as well as for the reporting and diagnosis of unusual events that may impact the future health of oceans, lakes, wildlife, and humans.


Postnatal expression of thalamic GABAA receptor subunits in the stargazer mouse model of absence epilepsy.
Absence seizures are known to originate from disruptions within the corticothalamocortical network; however, the precise underlying cellular and molecular mechanisms that induce hypersynchronicity and hyperexcitability are debated and likely to be complex and multifactorial. Recent studies implicate impaired thalamic GABAergic inhibition as a common feature in multiple animal models of absence epilepsy, including the well-established stargazer mouse model. Recently, we demonstrated region-specific increases in the whole tissue and synaptic levels of GABAA receptor (GABAAR) subunits α1 and β2, within the ventral posterior region of the thalamus in adult epileptic stargazer mice compared with nonepileptic control littermates. The objective of this study was to investigate whether such changes in GABAAR subunits α1 and β2 can be observed before the initiation of seizures, which occur around postnatal (PN) days 16-18 in stargazers. Semiquantitative western blotting was used to analyze the relative tissue level expression of GABAAR α1 and β2 subunits in the thalamus of juvenile stargazer mice compared with their nonepileptic control littermates at three different time points before the initiation of seizures. We show that there is a statistically significant increase in the expression of α1 and β2 subunits in the thalamus of stargazer mice, at the PN7-9 stage, compared with the control littermates, but not at PN10-12 and PN13-15 stages. These results suggest that an aberrant expression of GABAAR subunits α1 and β2 in the stargazers does not occur immediately before seizure onset and therefore is unlikely to directly contribute to the initiation of absence seizures.


Targeted Enrichment of rRNA Gene Tandem Arrays for Ultra-Long Sequencing by Selective Restriction Endonuclease Digestion.
Large regions of nearly identical repeats, such as the 45S ribosomal RNA (rRNA) genes of Nucleolus Organizer Regions (NORs), can account for major gaps in sequenced genomes. To assemble these regions, ultra-long sequencing reads that span multiple repeats have the potential to reveal sets of repeats that collectively have sufficient sequence variation to unambiguously define that interval and recognize overlapping reads. Because individual repetitive loci typically represent a small proportion of the genome, methods to enrich for the regions of interest are desirable. Here we describe a simple method that achieves greater than tenfold enrichment of Arabidopsis thaliana 45S rRNA gene sequences among ultra-long Oxford Nanopore Technology sequencing reads. This method employs agarose-embedded genomic DNA that is subjected to restriction endonucleases digestion using a cocktail of enzymes predicted to be non-cutters of rRNA genes. Most of the genome is digested into small fragments that diffuse out of the agar plugs, whereas rRNA gene arrays are retained. In principle, the approach can also be adapted for sequencing other repetitive loci for which gaps exist in a reference genome.


Delivered relative dose intensity in adolescent and young adult germ cell tumours in England: Assessment of data quality and consistency from clinical trials compared to national cancer registration data.
Adolescent and young adults (AYA) with germ cell tumours (GCT) have poorer survival rates than children and many older adults with the same cancers. There are several likely contributing factors to this, including the treatment received. The prognostic benefit of intended dose intensity is well documented in GCT from trials comparing regimens. However, evidence specific to AYA is limited by poor recruitment of AYA to trials and dose delivery outside trials not being well examined. We examined the utility of cancer registration data and a clinical trials dataset to investigate the delivery of relative dose intensity (RDI) in routine National Health Service practice in England, compared to within international clinical trials. Linked data from the Cancer Outcomes and Services Dataset (COSD) and the Systemic Anti-Cancer Therapy (SACT) dataset, and data from four international clinical trials were analysed. Survival over time was described using Kaplan-Meier estimation; overall, by age category, International Germ-Cell Cancer Collaborative Group (IGCCCG) classification, stage, tumour subtype, primary site, ethnicity and deprivation. Cox regression models were used to determine the fully adjusted effect of RDI on mortality risk. The quality of both datasets was critically evaluated and clinically enhanced. RDI was found to be well maintained in all datasets with higher RDIs associated with improved survival outcomes. Real-world data demonstrated several strengths, including population coverage and inclusion of sociodemographic variables and comorbidity. It is limited in GCT however, by the poor completion of data items enabling risk classification of patients and a higher proportion of missing data.


[Evaluation of the 2.5 mg fentanyl patch, applied using the half-side application procedure in patients with cancer pain].
Occasionally, pain control with the fentanyl patch may lead to overdose at an initial dose of 2.5 mg, as well as during dose increase from 2.5 to 5.0 mg. Respiratory depression and other adverse drug reactions associated with fentanyl overdose have been observed in several of our patients. We developed a procedure for applying one-half of the fentanyl patch formulations and evaluated the new mode of application by examining the fentanyl concentration in 32 patients with cancer-related pain who had been using the fentanyl patch for pain control. While some patients were treated with the full-sized 2.5-, 5.0-, or 7.5-mg formulations, others were treated with the half-sized 2.5-mg formulation. The fentanyl patch was equally divided by drawing a line on the side on which the product name and dose were written. Tegaderm was applied to the patient's skin, and after detaching from the protective liner, half of the patch was applied to overlap Tegaderm along the line and the other half applied directly to the skin. Blood samples were collected 48-72 h after patch application. The mean serum concentration of fentanyl given in the half-sized 2.5-mg formulation was 0.286 ng/ml, which was approximately one-half of the concentration of the full-sized 2.5-mg formulation, 0.544 ng/ml. Therefore the 2.5-mg fentanyl patch, applied using the one-half procedure we developed, is clinically useful.


Reliability and validity of different methods of estimating the one-repetition maximum during the free-weight prone bench pull exercise.
This study examined the reliability and validity of three methods of estimating the one-repetition maximum (1RM) during the free-weight prone bench pull exercise. Twenty-six men (22 rowers and four weightlifters) performed an incremental loading test until reaching their 1RM, followed by a set of repetitions-to-failure. Eighteen participants were re-tested to conduct the reliability analysis. The 1RM was estimated through the lifts-to-failure equations proposed by Lombardi and O'Connor, general load-velocity (L-V) relationships proposed by Sánchez-Medina and Loturco and the individual L-V relationships modelled using four (multiple-point method) or only two loads (two-point method). The direct method provided the highest reliability (coefficient of variation [CV] = 2.45% and intraclass correlation coefficient [ICC] = 0.97), followed by the Lombardi's equation (CV = 3.44% and ICC = 0.94), and no meaningful differences were observed between the remaining methods (CV range = 4.95-6.89% and ICC range = 0.81-0.91). The lifts-to-failure equations overestimated the 1RM (3.43-4.08%), the general L-V relationship proposed by Sánchez-Medina underestimated the 1RM (-3.77%), and no significant differences were observed for the remaining prediction methods (-0.40-0.86%). The individual L-V relationship could be recommended as the most accurate method for predicting the 1RM during the free-weight prone bench pull exercise.


Hormone receptors and other prognostic factors in breast cancer in Cuba.
Clinical management of breast cancer, making a prognosis and deciding on treatment, currently depend on defining prognostic factors, especially hormone receptors (HR). In addition to confirming the heterogeneity of the disease, these biological parameters are indispensable tools for designing personalized treatment. In this study, 1509 tumors from Cuban women diagnosed with breast cancer were examined. Hormone receptor (HR) expression was determined and correlated with a group of prognostic factors, such as age, tumor size, histological type, nuclear grade, histological grade, number of metastatic axillary lymph nodes, and clinical stage. Estrogen receptor (ER) expression was associated with low nuclear grade and histological grade, and with smaller tumor size (p < 0.05). Analysis of age at the time of diagnosis showed that ER expression was greater in patients in the group aged >50 years (p < 0.05). In general, ER expression was greater in patients in earlier clinical stages (p < 0.05). With regard to HR expression, 53% of tumors in this sample were ER+ and 49% were PR+. In 38% of cases, both receptors were positive and in 28% both receptors proved negative. The ER+/PR- combination was observed in 23% of cases while only 11% exhibited the ER-/PR+ combination. These findings indicate that approximately 72% of the tumors studied expressed some level of hormone dependency. This is the first report of HR expression in Cuba using immunohistochemical techniques and a representative sample of breast tumors diagnosed in different provinces around the country.


Structural equation modeling for hypertension and type 2 diabetes based on multiple SNPs and multiple phenotypes.
Genome-wide association studies (GWAS) have been successful in identifying genetic variants associated with complex diseases. However, association analyses between genotypes and phenotypes are not straightforward due to the complex relationships between genetic and environmental factors. Moreover, multiple correlated phenotypes further complicate such analyses. To resolve this complexity, we present an analysis using structural equation modeling (SEM). Unlike current methods that focus only on identifying direct associations between diseases and genetic variants such as single-nucleotide polymorphisms (SNPs), our method introduces the effects of intermediate phenotypes, which are related phenotypes distinct from the target, into the systematic genetic study of diseases. Moreover, we consider multiple diseases simultaneously in a single model. The procedure can be summarized in four steps: 1) selection of informative SNPs, 2) extraction of latent variables from the selected SNPs, 3) investigation of the relationships among intermediate phenotypes and diseases, and 4) construction of an SEM. As a result, a quantitative map can be drawn that simultaneously shows the relationship among multiple SNPs, phenotypes, and diseases. In this study, we considered two correlated diseases, hypertension and type 2 diabetes (T2D), which are known to have a substantial overlap in their disease mechanism and have significant public health implications. As intermediate phenotypes for these diseases, we considered three obesity-related phenotypes-subscapular skin fold thickness, body mass index, and waist circumference-as traits representing subcutaneous adiposity, overall adiposity, and abdominal adiposity, respectively. Using GWAS data collected from the Korea Association Resource (KARE) project, we applied the proposed SEM process. Among 327,872 SNPs, 24 informative SNPs were selected in the first step (p<1.0E-05). Ten latent variables were generated in step 2. After an exploratory analysis, we established a path diagram among phenotypes and diseases in step 3. Finally, in step 4, we produced a quantitative map with paths moving from specific SNPs to hypertension through intermediate phenotypes and T2D. The resulting model had high goodness-of-fit measures (χ2 = 536.52, NFI = 0.997, CFI = 0.998, GFI = 0.995, AGFI = 0.993, RMSEA = 0.012).


A measure of quality of life after abdominal surgery.
To develop a reliable and valid measure of short-term quality of life after abdominal surgery. A major limitation of clinical trials evaluating laparoscopic surgical procedures has been the lack of a measure of short-term quality of life after abdominal surgery. We used existing health status measures, focus groups, and semi-structured patient interviews to generate a prototype questionnaire of 51 items, which was administered to patients within 2 weeks after an abdominal surgical procedure. We used structural equations modeling to reduce the number of items, retaining the three items with the highest factor loadings on each of the factors that accounted for one or more eigenvalue. We administered the prototype questionnaire to 500 patients (mean age [SD] 53.4 [16.0], 51.4% male, 73.0% inpatient) at a mean 4.1 days after an abdominal surgical procedure. Item reduction yielded an 18-item measure with 6 sub-scales. The final instrument demonstrated good model fit in relation to our hypothesized factors (root mean square error of approximation 0.085, goodness-of-fit index 0.89). We developed a reliable and valid 18-item, 6-subscale measure of health-related quality of life after abdominal surgery, for use as an outcome measure in studies comparing laparoscopic and conventional abdominal surgery.


Incidence of Type 1 Diabetes in Relation to Exposure to Rotavirus Infections in Pre- and Postvaccine Birth Cohorts in Finland.
To explore the incidence of type 1 diabetes in children in relation to exposure to rotavirus infections. A nationwide register-based ecological study on the 1995-2015 birth cohorts in Finland compared those born before and after the national implementation of the rotavirus vaccine in 2009. When the prevaccine 2001-2005 birth cohorts were compared with the postvaccine birth cohorts, the number of children exposed to rotavirus infection by the age of 5 years decreased from 2,522 per 100,000 children (2.5%) to 171 per 100,000 children (0.2%), while the incidence of type 1 diabetes in those aged <5 years decreased from 71.5 to 54.4 per 100,000 person-years (incidence rate ratio 0.79, 95% CI 0.71-0.86). At the population level, a decrease in exposure to rotavirus infections was associated with a decrease in the incidence of type 1 diabetes in young children.


Elucidating the coordination chemistry of the radium ion for targeted alpha therapy.
The coordination chemistry of Ra2+ is poorly defined, hampering efforts to design effective chelators for 223Ra-based targeted alpha therapy. Here, we report the complexation thermodynamics of Ra2+ with the biomedically-relevant chelators DOTA and macropa. Our work reveals the highest affinity chelator to date for Ra2+ and advances our understanding of key factors underlying complex stability and selectivity for this underexplored ion.


Cooperative motion of spheres arranged in periodic grids between two parallel walls.
In this article, we analyze the collective motion of a two-dimensional periodic array of spheres in a slit-pore confined by two parallel planar walls. We determine the friction coefficient of the spheres when all particles move with the same velocity along a particular direction and cooperate with each other in their motion. In order to solve this many-body problem, we use Stokesian dynamics algorithm and resolve multiparticle hydrodynamic interactions in wall-bounded geometry. Apart from particle-particle interactions, we also recognize that the aforementioned collective motion of all particles creates a cumulative effect on the fluid medium. This effect is manifested as either a net induced flow for a periodic pressure field or an additional pressure gradient for quiescent fluid. In our analysis, we focus on both periodic pressure and no-flow conditions. For both cases, the hydrodynamic friction on the translating particles is calculated using our multiparticle Stokesian dynamics simulation. The simulation for the no-flow condition is relatively straightforward-we only need to compute the multiparticle hydrodynamic interactions in quiescent fluid. However, for the periodic pressure condition, the net induced flow dragged by the particles has to be evaluated also. We express this net induced flow in terms of an additional pressure-driven velocity field. We present the hydrodynamic friction as a function of the dimensions of the two-dimensional periodic lattice. For closely packed arrays, the results show a considerable reduction in friction coefficients that usually increase with interparticle distance. Hence, our work renders the theoretical justification for other recent findings that indicate the importance of interparticle mutual cooperation.


Failure of cement-augmented pedicle screws in the osteoporotic spine: a case report.
The treatment of patients with osteoporosis and spinal abnormalities that require surgical intervention is difficult because of the challenge of achieving fixation in osteoporotic bone. As the population ages, this challenge is becoming a common problem in the field of spinal surgery. Although numerous publications exist about the biomechanical benefits of various fixation devices and techniques, no standard of care has emerged that offers a clear method for accomplishing spinal stabilization in such patients. This case presents the failure mode of cement-augmented pedicle screws in a patient with severe osteoporosis, a description of the methods used to attain fixation and spinal stability during the revision surgery, and the outcome achieved for the patient 1 year after surgery. An 82-year-old female with a T9 burst fracture and a history of osteoporosis underwent minimally invasive instrumentation from T5 to T12, fusion from T7 to T11, and decompression from T8 to T10. Four weeks after surgery, the patient returned to the hospital because of back pain. Imaging studies showed that the pedicle screws at T11 and T12, which were augmented with polymethylmethacrylate (PMMA), had pulled out of the vertebral bodies. The pedicle screws failed by disengaging from the PMMA and displacing posteriorly and inferiorly. The PMMA did not appear to move during this process. A revision surgery was performed, in which the posterior construct was extended caudally and cephalad, the pedicle screws were augmented with PMMA, and a titanium hook and woven polyester band were used to increase the points of fixation. At 1-year follow-up after revision, our patient showed radiographic evidence of fusion, and the construct continued to maintain stability in the osteoporotic spine.


Generation of Nanog reporter mice that distinguish pluripotent stem cells from unipotent primordial germ cells.
Nanog is a core transcription factor specifically expressed not only in the pluripotent stem cells (PSCs), such as embryonic stem cells (ESCs), embryonic germ cells (EGCs), and induced PSCs (iPSCs), but also in the unipotent primordial germ cells (PGCs). Although Nanog promoter/enhancer regions are well characterized by in vitro analyses, direct correlations between the regulatory elements for Nanog expression and in vivo expression patterns of Nanog have not been fully clarified. In this study, we generated Nanog-RFP transgenic (Tg) mice in which expression of red fluorescent protein (RFP) is driven by a 5.2 kb Nanog promoter/enhancer region. As expected, RFP was expressed in the inner cell mass of blastocysts, ESCs, and iPSCs. However, RFP fluorescence was not observed in PGCs, although Nanog was expressed in PGCs. Because RFP fluorescence was visible in the PGC-derived pluripotent EGCs in culture, it was suggested that the reporter gene expression was specifically activated in PSCs. In conclusion, we have generated a novel Nanog-RFP Tg mouse line that can selectively tag PSCs over unipotent PGCs.


Chronic Hiccups: An Underestimated Problem.
Persistent singultus, hiccupping that lasts for longer than 48 hours, can have a tremendous impact on a patient's quality of life. Although involved neurologic structures have been identified, the function of hiccups remains unclear-they have been controversially interpreted as a primitive reflex preventing extent swallowing of amniotic fluid in utero, an archaic gill ventilation pattern, or a fetus' preparation for independent breathing. Persistent singultus often presents as a symptom for various diseases, most commonly illnesses of the central nervous system or gastrointestinal tract; they can also be evoked by a variety of pharmacological agents. It is often impossible to define a singular cause. A wide range of treatment attempts, pharmacological and nonpharmacological, have been concerted to this date; however, chlorpromazine remains the only Food and Drug Administration-approved drug in this context. Large-scale studies on efficacy and tolerance of other therapeutic strategies are lacking. Gabapentin, baclofen, and metoclopramide have been reported to accomplish promising results in reports on the therapy of persistent singultus; they may also be effective when given in combination with other drugs, eg, proton pump inhibitors, or as conjoined therapy. As another approach of note, acupuncture treatment was able to abolish hiccups in a number of studies. When managing hiccup patients within the clinical routine, it is of importance to conduct a comprehensive and effective diagnostic workup; a well-functioning interdisciplinary team is needed to address possible causes for the symptom. Persistent singultus is a medical problem not to be underestimated; more research on options for effective treatment would be greatly needed.


Sodium MRI of the human heart at 7.0 T: preliminary results.
The objective of this work was to examine the feasibility of three-dimensional (3D) and whole heart coverage (23)Na cardiac MRI at 7.0 T including single-cardiac-phase and cinematic (cine) regimes. A four-channel transceiver RF coil array tailored for (23)Na MRI of the heart at 7.0 T (f = 78.5 MHz) is proposed. An integrated bow-tie antenna building block is used for (1)H MR to support shimming, localization and planning in a clinical workflow. Signal absorption rate simulations and assessment of RF power deposition were performed to meet the RF safety requirements. (23) Na cardiac MR was conducted in an in vivo feasibility study. 3D gradient echo (GRE) imaging in conjunction with Cartesian phase encoding (total acquisition time T(AQ) = 6 min 16 s) and whole heart coverage imaging employing a density-adapted 3D radial acquisition technique (T(AQ) = 18 min 20 s) were used. For 3D GRE-based (23)Na MRI, acquisition of standard views of the heart using a nominal in-plane resolution of (5.0 × 5.0) mm(2) and a slice thickness of 15 mm were feasible. For whole heart coverage 3D density-adapted radial (23)Na acquisitions a nominal isotropic spatial resolution of 6 mm was accomplished. This improvement versus 3D conventional GRE acquisitions reduced partial volume effects along the slice direction and enabled retrospective image reconstruction of standard or arbitrary views of the heart. Sodium cine imaging capabilities were achieved with the proposed RF coil configuration in conjunction with 3D radial acquisitions and cardiac gating. Cardiac-gated reconstruction provided an enhancement in blood-myocardium contrast of 20% versus the same data reconstructed without cardiac gating. The proposed transceiver array enables (23)Na MR of the human heart at 7.0 T within clinical acceptable scan times. This capability is in positive alignment with the needs of explorations that are designed to examine the potential of (23)Na MRI for the assessment of cardiovascular and metabolic diseases.


Drug Repurposing Strategy (DRS): Emerging Approach to Identify Potential Therapeutics for Treatment of Novel Coronavirus Infection.
Drug repurposing is also termed as drug repositioning or therapeutic switching. This method is applied to identify the novel therapeutic agents from the existing FDA approved clinically used drug molecules. It is considered as an efficient approach to develop drug candidates with new pharmacological activities or therapeutic properties. As the drug discovery is a costly, time-consuming, laborious, and highly risk process, the novel approach of drug repositioning is employed to increases the success rate of drug development. This strategy is more advantageous over traditional drug discovery process in terms of reducing duration of drug development, low-cost, highly efficient and minimum risk of failure. In addition to this, World health organization declared Coronavirus disease (COVID-19) as pandemic globally on February 11, 2020. Currently, there is an urgent need to develop suitable therapeutic agents for the prevention of the outbreak of COVID-19. So, various investigations were carried out to design novel drug molecules by utilizing different approaches of drug repurposing to identify drug substances for treatment of COVID-19, which can act as significant inhibitors against viral proteins. It has been reported that COVID-19 can infect human respiratory system by entering into the alveoli of lung via respiratory tract. So, the infection occurs due to specific interaction or binding of spike protein with angiotensin converting enzyme-2 (ACE-2) receptor. Hence, drug repurposing strategy is utilized to identify suitable drugs by virtual screening of drug libraries. This approach helps to determine the binding interaction of drug candidates with target protein of coronavirus by using computational tools such as molecular similarity and homology modeling etc. For predicting the drug-receptor interactions and binding affinity, molecular docking study and binding free energy calculations are also performed. The methodologies involved in drug repurposing can be categorized into three groups such as drug-oriented, target-oriented and disease or therapy-oriented depending on the information available related to quality and quantity of the physico-chemical, biological, pharmacological, toxicological and pharmacokinetic property of drug molecules. This review focuses on drug repurposing strategy applied for existing drugs including Remdesivir, Favipiravir, Ribavirin, Baraticinib, Tocilizumab, Chloroquine, Hydroxychloroquine, Prulifloxacin, Carfilzomib, Bictegravir, Nelfinavir, Tegobuvir and Glucocorticoids etc to determine their effectiveness toward the treatment of COVID-19.


Airway responses to methacholine and exercise at high altitude in healthy lowlanders.
Peribronchial edema has been proposed as a mechanism enhancing airway responses to constrictor stimuli. Acute exposure to altitude in nonacclimatized lowlanders leads to subclinical interstitial pulmonary edema that lasts for several days after ascent, as suggested by changes in lung mechanics. We, therefore, investigated whether changes in lung mechanics consistent with fluid accumulation at high altitude within the lungs are associated with changes in airway responses to methacholine or exercise. Fourteen healthy subjects were studied at 4,559 and at 120 m above sea level. At high altitude, both static and dynamic lung compliances and respiratory reactance at 5 Hz significantly decreased, suggestive of interstitial pulmonary edema. Resting minute ventilation significantly increased by approximately 30%. Compared with sea level, inhalation of methacholine at high altitude caused a similar reduction of partial forced expiratory flow but less reduction of maximal forced expiratory flow, less increments of pulmonary resistance and respiratory resistance at 5 Hz, and similar effects of deep breath on pulmonary and respiratory resistance. During maximal incremental exercise at high altitude, partial forced expiratory flow gradually increased with the increase in minute ventilation similarly to sea level but both achieved higher values at peak exercise. In conclusion, airway responsiveness to methacholine at high altitude is well preserved despite the occurrence of interstitial pulmonary edema. We suggest that this may be the result of the increase in resting minute ventilation opposing the effects and/or the development of airway smooth muscle force, reduced gas density, and well preserved airway-to-parenchyma interdependence.


Novel equation to determine the hepatic triglyceride concentration in humans by MRI: diagnosis and monitoring of NAFLD in obese patients before and after bariatric surgery.
Non-alcoholic fatty liver disease (NAFLD) is caused by abnormal accumulation of lipids within liver cells. Its prevalence is increasing in developed countries in association with obesity, and it represents a risk factor for non-alcoholic steatohepatitis (NASH), cirrhosis and hepatocellular carcinoma. Since NAFLD is usually asymptomatic at diagnosis, new non-invasive approaches are needed to determine the hepatic lipid content in terms of diagnosis, treatment and control of disease progression. Here, we investigated the potential of magnetic resonance imaging (MRI) to quantitate and monitor the hepatic triglyceride concentration in humans. A prospective study of diagnostic accuracy was conducted among 129 consecutive adult patients (97 obesity and 32 non-obese) to compare multi-echo MRI fat fraction, grade of steatosis estimated by histopathology, and biochemical measurement of hepatic triglyceride concentration (that is, Folch value). MRI fat fraction positively correlates with the grade of steatosis estimated on a 0 to 3 scale by histopathology. However, this correlation value was stronger when MRI fat fraction was linked to the Folch value, resulting in a novel equation to predict the hepatic triglyceride concentration (mg of triglycerides/g of liver tissue = 5.082 + (432.104 * multi-echo MRI fat fraction)). Validation of this formula in 31 additional patients (24 obese and 7 controls) resulted in robust correlation between the measured and estimated Folch values. Multivariate analysis showed that none of the variables investigated improves the Folch prediction capacity of the equation. Obese patients show increased steatosis compared to controls using MRI fat fraction and Folch value. Bariatric surgery improved MRI fat fraction values and the Folch value estimated in obese patients one year after surgery. Multi-echo MRI is an accurate approach to determine the hepatic lipid concentration by using our novel equation, representing an economic non-invasive method to diagnose and monitor steatosis in humans.


Tracing pathway activities with kinase inhibitors and reverse phase protein arrays.
Controlling aberrant protein kinase activity is a promising strategy for a variety of diseases, particularly cancer. Hence, the development of kinase inhibitors is currently a focal point for pharmaceutical research. In this study we utilize a chip-based reverse phase protein array (RPA) platform for profiling of kinase inhibitors in cell-based assays. In combination with the planar wave-guide technology the assay system has an absolute LOD down to the low zeptomole range. A431 cell lysates were analyzed for the activation state of key effectors in the epidermal growth factor (EGF) and insulin signaling pathways to validate this model for compound screening. A microtiter-plate format for growing, treating, and lysing cells was shown to be suitable for this approach, establishing the value of the technology as a screening tool for characterization of large numbers of kinase inhibitors against a wide variety of cellular signaling pathways. Moreover, the reverse array format allows rapid development of site-specific phosphorylation assays, since in contrast to ELISA type systems only a single antigen-specific antibody is required.


Evaluation of microbial reduction of Fe(III)EDTA in a chemical absorption-biological reduction integrated NOx removal system.
A chemical absorption-biological reduction integrated process can be used to remove nitrogen oxides (NOx) from flue gas. In such a process, nitric oxide (NO) can be effectively absorbed by the ferrous chelate of ethylenediaminetetraacetate (Fe(II)EDTA) to form Fe(II)EDTA-NO, which can be biologically regenerated by denitrifying bacteria. However, in the course of these processes, part of the Fe(II)EDTA is also oxidized to Fe(III)EDTA. The reduction of Fe(III)EDTA to Fe(II)EDTA depends on the activity of iron-reducing bacteria in the system. Therefore, the effectiveness of the system relies on how to effectively bioreduce Fe(III)EDTA and Fe(II)EDTA-NO in the system. In this paper, a strain identified as Escherichia coli FR-2 (iron-reducing bacterium) was used to investigate the reduction rate of Fe(III)EDTA. The experimental results indicate that Fe(III)EDTA-NO and Fe(II)EDTA in the system can inhibit both the FR-2 cell growth and thus affect the Fe(III)EDTA reduction. The FR-2 cell growth rate and Fe(III)EDTA reduction rate decreased with increasing Fe(II)EDTA-NO and Fe(II)EDTA concentration in the solution. When the concentration of Fe(II)EDTA-NO reached 3.7 mM, the FR-2 cell growth almost stopped. A mathematical model was developed to explain the cell growth and inhibition kinetics. The predicted results are close to the experimental data and provide a preliminary evaluation of the kinetics of the biologically mediated reactions necessary to regenerate the spent scrubber solution.


Perception of Medical Assistance in Dying Among Asian Buddhists Living in Montreal, Canada.
In the Western world including Canada, grievous and irredeemable health conditions, which cause unbearable suffering, has given support to the legalization of medical aid in dying (MAiD). It is unknown how Asian Buddhists who are in contact with the Western culture perceive MAiD. In this qualitative study, 16 Asian Buddhists living in Montreal took part in a semi-structured interview. Contrary to general findings in the literature, religious affiliation do not always determine moral stances and practical decisions when it comes to MAiD. Some participants were willing to take some freedom with the doctrine and based their approval of MAiD on the right to self-determination. Those who disapproved the use of MAiD perceived it as causing unnatural death, creating bad karma, and interfering with a conscious death. End-of-life (EoL) care providers have to remain sensitive to each patient's spiritual principles and beliefs to understand their needs and choices for EoL care.


Preoperative Predictors of Level of Difficulty of Laparoscopic Cholecystectomy.
Laparoscopic cholecystectomy (LC) is the gold standard treatment for symptomatic gallbladder stone disease. This is due to its safety, reliability, cost-effectiveness, negligible mortality, shorter duration of hospitalization (early return to work), better cosmesis, minimal wound complications, and temporary paralytic ileus. In spite of these, conversion to open cholecystectomy which is sometimes required in difficult cases could be challenging. The aim of the present study is to aid the prediction of difficult cases undergoing LC, thereby better selection of patients with the least conversion rates. This prospective study was conducted on 100 consecutive patients with cholecystitis, over a 2-year period from January 1, 2017, to December 31, 2018, having undergone LC. Various preoperative parameters, including age, sex, previous attacks of cholecystitis, deranged liver functions, and ultrasonographic findings, were analyzed for their effects for predicting the level of difficulty during LC. Twenty-five percent of the cases were correctly predicted as difficult in the age group of >65 years. Cholecystitis was more common (79%) in females, but difficulties were encountered more frequently while performing LC in males. Abnormal serum hepatic and pancreatic enzyme profiles were associated with difficulties during surgery as about 83.3% of the patients predicted as moderately difficult peroperatively had deranged liver functions. The preoperative ultrasonography findings were helpful for predicting the degree of difficulty involved in the procedure. About 33.3% of the patients that had pericholecystic fluid on ultrasound preoperatively were correctly predicted to have moderately difficult surgeries. The Chi-square test and P value were used to determine statistical significance. Females, the absence of previous repeated attacks of cholecystitis and hospitalizations, no upper abdominal surgery in the past, normal liver function tests, normal amylase levels, nondistended and uncontracted gallbladder, absence of pericholecystic collection, afebrile, and single stone are positive preoperative predictors of safe LC in symptomatic gallbladder stone disease.


Light guiding in a slot waveguide that includes an additional confining core region.
We present a design of a slot waveguide in which the core layer is orthogonally slotted to form a rectangular sub-core. While the overall guiding and coupling efficiency remains the same as a conventional slot waveguide, the field confinement is enhanced and appears two-dimensional. The waveguiding is controllable by selecting the intermediate index as well as various geometrical parameters. In addition, by changing different variables, the linear/nonlinear dispersion and birefringence can be tailored with extended ranges. Constant-dispersion points, where the dispersion is insensitive to size changes, are also demonstrated.


Cochlear Dysfunction Is a Frequent Feature of Facioscapulohumeral Muscular Dystrophy Type 1 (FSHD1).
Facioscapulohumeral muscular dystrophy type 1 (FSHD) represents one of the most common forms of muscular hereditary diseases and it is characterized by a great clinical variability with the typical muscular symptoms and other clinical features, including hearing impairment. However, etiopathogenetic mechanisms of auditory dysfunction are still not completely understood and it has been suggested that it could be assigned to a cochlear alteration that is present even in those subjects with a normal pure tonal audiometry (PTA) examination. We found out the cochlear function in 26 patients with molecular diagnosis of FSHD1 and in healthy controls. All patients underwent complete neurological and audiological examinations, including FSHD clinical score, pure-tone audiometry (PTA), and otoacoustic emissions (OAEs), in particular transient evoked otoacoustic emissions (TEOAEs) and distortion product evoked otoacoustic emissions (DPOAEs). All FSHD1 patients showed significantly reduced DPOAEs and TEOAEs, bilaterally and at all frequencies, even when considering only subjects with a normal PTA or a mild muscular involvement (FSHD score ≤ 2). No correlation between OAEs and FSHD clinical score was found. Cochlear echoes represent a sensitive tool in detecting subclinical cochlear dysfunction in FSHD1 even in subjects with normal hearing and/or subtle muscle involvement. Our study is focused on the importance of evaluating the cochlear alteration through OAEs and, in particular, by performing TEOAEs and DPOAEs sequentially, to evaluate more frequent specificities of cochlear dysfunction with a wider spectrum of analysis.


The Beta Lactam Antibiotics as an Empirical Therapy in a Developing Country: An Update on Their Current Status and Recommendations to Counter the Resistance against Them.
In a developing country like India, where the patients have to bear the cost of their healthcare, the microbiological culture and the sensitivity testing of each and every infection is not feasible. Moreover, there are lacunae in the data storage, management and the sharing of knowledge with respect to the microorganisms which are prevalent in the local geographical area and with respect to the antibiotics which are effective against them. Thus, an empirical therapy for treating infections is imperative in such a setting. The beta lactam antibiotics have been widely used for the empirical treatment of infections since the the discovery of penicillin. Many generations of beta lactams have been launched with, the claims of a higher sensitivity and less resistance, but their sensitivity has drastically decreased over time. Thus, the preference for beta lactams, especially the cephalosporins, as an empirical therapy, among the prescribers was justified initially, but the current sensitivity patterns do not support their empirical use in hospital and community acquired infections. There is a need for increasing the awareness and the attitudinal change among the prescribers, screening of the antibiotic prescriptions, the strict implementation of antibiotic policies in hospital settings, restricting the hospital supplies and avoiding the prescriptions of beta lactams, a regular census of the local sensitivity patterns to formulate and update the antibiotic policies, upgradation of the laboratory facilities for a better and faster detection of the isolates, proper collection, analyses and sharing of the data and the encouragement of the research and development of newer antibiotics with novel mechanisms of action.


Properties of triiodothyronine-binding proteins in liver cytosol of rat.
The electrophoretic mobility and the sedimentation coefficient were determined in partially purified preparations of both rat liver cytosol and serum triiodothyronine (T3)-binding proteins. Crude cytosol and serum, each labeled with [125I]T3, were filtered through a Sephadex G-100 column. The cytosol yielded a single T3-binding peak, whereas three binding components were recognized in the serum. Protamine sulfate precipitated the cytosol T3-binding protein, but had no effect on the serum T3-binders. The cytosol protein and the three binding proteins from serum were analyzed by polyacrylamide gel electrophoresis and sucrose density gradient centrifugation. The cytosol binder migrated as a single peak on gel electrophoresis with an Rf of 0.53, whereas the serum proteins had RfS between 0.27-0.33. The sedimentation coefficient of the cytosol protein was 6.3 S, whereas it was 4.1 S for the major binding protein of the serum. These data indicate that: (i) preliminary purification by gel chromatography is a useful step for better characterization of the T3-binding proteins of the cytosol and serum; (ii) the cytosol binder is an acidic protein with completely different properties from those of the serum T3-binding proteins.


A loss-of-function mutant allele of a glycosyl hydrolase gene has been co-opted for seed weight control during soybean domestication.
The resultant DNA from loss-of-function mutation can be recruited in biological evolution and development. Here, we present such a rare and potential case of "to gain by loss" as a neomorphic mutation during soybean domestication for increasing seed weight. Using a population derived from a chromosome segment substitution line of Glycine max (SN14) and Glycine soja (ZYD06), a quantitative trait locus (QTL) of 100-seed weight (qHSW) was mapped on chromosome 11, corresponding to a truncated β-1, 3-glucosidase (βGlu) gene. The novel gene hsw results from a 14-bp deletion, causing a frameshift mutation and a premature stop codon in the βGlu. In contrast to HSW, the hsw completely lost βGlu activity and function but acquired a novel function to promote cell expansion, thus increasing seed weight. Overexpressing hsw instead of HSW produced large soybean seeds, and surprisingly, truncating hsw via gene editing further increased the seed size. We further found that the core 21-aa peptide of hsw and its variants acted as a promoter of seed size. Transcriptomic variation in these transgenic soybean lines substantiated the integration hsw into cell and seed size control. Moreover, the hsw allele underwent selection and expansion during soybean domestication and improvement. Our work cloned a likely domesticated QTL controlling soybean seed weight, revealed a novel genetic variation and mechanism in soybean domestication, and provided new insight into crop domestication and breeding, and plant evolution.


Systematic literature review and network meta-analysis comparing bone-targeted agents for the prevention of skeletal-related events in cancer patients with bone metastasis.
Complications from skeletal-related events (SREs) constitute a challenge in the care of cancer patients with bone metastasis (BM). This study evaluated the comparative effectiveness of pamidronate, ibandronate, zoledronate, and denosumab in reducing the morbidity of SREs in cancer patients with BM. Medline (1948 to January 2014), Embase (1980 to January 2014), the Cochrane Library (2014 issue 1), and Web of Science with Conference Proceedings (1970 to January 2014) were searched. Only randomized controlled trials assessing denosumab, bisphosphonates, or placebo in cancer patients with BM were included. The primary outcomes were SREs and SREs by type. The network meta-analysis (NMA) was performed with a random-effects Bayesian model. The NMA included 14 trials with 10,192 patients. Denosumab was superior to placebo in reducing the risk of SREs (odds ratio [OR]: 0.49; 95% confidence interval [CI]: 0.31-0.75), followed by zoledronate (OR: 0.57; 95% CI: 0.41-0.77) and pamidronate (OR: 0.55; 95% CI: 0.41-0.72). Ibandronate compared with placebo could not reduce the risk of SREs. Denosumab was superior to placebo in reducing the risk of pathologic fractures (OR: 0.50; 95% CI: 0.32-0.79), followed by zoledronate (OR: 0.61; 95% CI: 0.43-0.86). Denosumab was superior to placebo in reducing the risk of radiation (OR: 0.51; 95% CI: 0.35-0.75), followed by pamidronate (OR: 0.67; 95% CI: 0.52-0.86) and zoledronate (OR: 0.70; 95% CI: 0.52-0.96). This NMA showed that denosumab, zoledronate, and pamidronate were generally effective in preventing SREs in cancer patients with BM. Denosumab and zoledronate were also associated with reductions in the risk of pathologic fractures and radiation compared with placebo. Denosumab was shown to be the most effective of the bone-targeted agents.


CD109-GP130 interaction drives glioblastoma stem cell plasticity and chemoresistance through STAT3 activity.
Glioma stem cells (GSCs) drive propagation and therapeutic resistance of glioblastomas, the most aggressive diffuse brain tumors. However, the molecular mechanisms that maintain the stemness and promote therapy resistance remain poorly understood. Here we report CD109/STAT3 axis as crucial for the maintenance of stemness and tumorigenicity of GSCs and as a mediator of chemoresistance. Mechanistically, CD109 physically interacts with glycoprotein 130 to promote activation of the IL-6/STAT3 pathway in GSCs. Genetic depletion of CD109 abolished the stemness and self-renewal of GSCs and impaired tumorigenicity. Loss of stemness was accompanied with a phenotypic shift of GSCs to more differentiated astrocytic-like cells. Importantly, genetic or pharmacologic targeting of CD109/STAT3 axis sensitized the GSCs to chemotherapy, suggesting that targeting CD109/STAT3 axis has potential to overcome therapy resistance in glioblastoma.


Quantitative studies of single-cell properties in monkey striate cortex. II. Orientation specificity and ocular dominance.
1. Quantitative analyses of orientation specificity and ocular dominance were carried out in striate cortex of the rhesus monkey. 2. Sharpness of orientation selectivity was greater for simple (S type) than for complex (CX type) cells. CX-type cells became more broadly tuned in the deeper cortical layers: S-type cells were equally well tuned throughout the cortex. 3. Sharpness of orientation selectivity for S-type cells was similar at all retinal eccentricities studied (0 degrees - 20 degrees from the fovea):in CX-type cells orientation selectivity decreased slightly with increasing eccentricity. 4. The orientation tuning of binocular cells was similar when mapped separately through each eye. 5. Orientation selectivity and direction selectivity are independent of each other, suggesting that separate neural mechanisms give rise to them. 6. More CX-type cells can be binocularly activated than S-type cells (88% versus 49%). The ocular dominance of S-type cells is similar in all cortical layers: for CX-type cells there is an increase in the number of cells in ocular-dominance category 4 in layers 5 and 6.


Nephrology in Veterinary Medicine.
Veterinary nephrology is a specialized field of veterinary medicine providing a high level of care for animals with all types of kidney disease. Veterinarians complete extensive training to become board-certified in veterinary nephrology-urology. Companion animal nephrology is the most advanced field; however, all species are afflicted by a variety of renal disorders. Most naturally occurring animal kidney diseases have similar disorders found in people; where veterinary research is lacking, clinical management is often modified from standard of care in people. Veterinarians have become adept at scaling down procedures to safely perform them on dogs and cats weighing only a few kilograms. Advanced diagnostics (renal biopsy, cystoscopy, fluoroscopic studies, etc. ) and therapeutics (renal replacement therapy, interventional endourology, etc. ) are commonly performed within the practice of veterinary nephrology-urology. Collaboration between veterinary and human nephrologists may advance both disciplines and improve care for people and animals alike.


Evidence of coat color variation sheds new light on ancient canids.
We have used a paleogenetics approach to investigate the genetic landscape of coat color variation in ancient Eurasian dog and wolf populations. We amplified DNA fragments of two genes controlling coat color, Mc1r (Melanocortin 1 Receptor) and CBD103 (canine-β-defensin), in respectively 15 and 19 ancient canids (dogs and wolf morphotypes) from 14 different archeological sites, throughout Asia and Europe spanning from ca. 12 000 B.P. (end of Upper Palaeolithic) to ca. 4000 B.P. (Bronze Age). We provide evidence of a new variant (R301C) of the Melanocortin 1 receptor (Mc1r) and highlight the presence of the beta-defensin melanistic mutation (CDB103-K locus) on ancient DNA from dog-and wolf-morphotype specimens. We show that the dominant K(B) allele (CBD103), which causes melanism, and R301C (Mc1r), the variant that may cause light hair color, are present as early as the beginning of the Holocene, over 10,000 years ago. These results underline the genetic diversity of prehistoric dogs. This diversity may have partly stemmed not only from the wolf gene pool captured by domestication but also from mutations very likely linked to the relaxation of natural selection pressure occurring in-line with this process.


Recruitment of African Americans with chronic renal insufficiency into a multicenter clinical trial: the african american study of kidney disease and hypertension.
In patients with hypertensive nephrosclerosis, the African American Study of Kidney Disease and Hypertension (AASK) demonstrated the superiority of angiotensin-converting enzyme inhibitor therapy in blunting progression of renal disease compared with a b blocker and a dihydropyridine calcium channel blocker. In addition, the study found that a blood pressure treatment strategy that resulted in an achieved blood pressure of 128/78 mm Hg (low blood pressure goal) was no more effective in slowing the progression of renal disease than a strategy that resulted in a blood pressure of 141/85 mm Hg (usual blood pressure goal). AASK, which enrolled only African Americans with mild to moderate chronic renal insufficiency, also provided an opportunity to evaluate recruitment methods in minority populations. Eighty-three percent of patients were recruited through screening in clinical practice. To randomize 635 patients, 558,295 charts were reviewed (approximately 879 charts per randomized patient). More than half of the randomized patients (n=635 or 58%) were found by chart review. Sixty percent of women with creatinine levels considered within the normal range had at least mild chronic renal insufficiency. Screening in clinical practice was the most effective strategy to recruit participants with mild to moderate chronic renal insufficiency and hypertension into the clinical trial. This technique may also be an effective approach in trials of other essentially asymptomatic conditions.


Cell association increases RPE outgrowth from primary explant.
Most studies of cell growth of the RPE employ cultures which have been previously passaged, but here we investigated freshly-explanted RPE cells, which may have been growth-quiescent for years, within the eye to determine whether co-culture affects initial outgrowth in primary culture. Bovine or human RPE were co-cultured in primary culture with several cell types to test the effects of homologous or heterologous cell association. For bovine RPE, cell number was measured over 14 days in cultures of RPE alone, or RPE in co-culture with irradiated living cells, with fixed-killed cells, with cells separated from the RPE by a semipermeable membrane, or in medium conditioned by the cell types used for co-culture. For human RPE, isolates from all donors were randomized, over a 13-month period, to co-culture or to culture alone. The number of cultures attaining a cell number at one month that was sufficient for further propagation was compared. Co-culture with irradiated living cells increased the growth of primary cultures of both bovine and human RPE. Living cells were required; fixed-killed cells were ineffective. The outgrowth-promoting activity was not tissue or species specific, and it appeared to require close cell association between the RPE and the co-culture cell population. Conditioned media were ineffective and rather were slightly growth inhibitory. Primary RPE cells showed an earlier expression of vimentin (a marker of Gzero-G1 transition), more rapid cell spreading, and a greater increase in cell number between 7 and 14 days after explant when grown in co-culture than when cultured alone. The results indicate that cell association between non-mitotic RPE cells and previously cultured cells of many types increases the outgrowth of the RPE by accelerating the early stages of growth activation in vitro. Co-culture methods offer a practical means for increasing the likelihood of producing cultures from small RPE isolates. Further, should cells involved in proliferative pathologies in situ associate with non-mitotic RPE within the monolayer, the latter cells may also be activated, leading to an augmentation of the pathology.


Development of a column switching for direct online enrichment and separation of polar and nonpolar analytes from aqueous matrices.
Trace substances in surface waters may threaten health and pose a risk for the aquatic environment. Moreover, separation and detection by instrumental analysis is challenging due to the low concentration and the wide range of polarities. Separation of polar and nonpolar analytes can be achieved by using stationary phases with different selectivity. Lower limits of detection of trace substances can be obtained by offline enrichment on solid phase materials. However, these practices require substantial effort and are time consuming and costly. Therefore, in this study, a column switching was developed to enrich and separate both polar and nonpolar analytes by an on-column large volume injection of aqueous samples. The column switching can significantly reduce the effort and time for analyzing trace substances without compromising on separation and detection. A reversed phase (RP) column is used to trap the nonpolar analytes. The polar analytes are enriched on a porous graphitized carbon column (PGC) coupled serially behind the RP column. A novel valve switching system is implemented to enable elution of the nonpolar analytes from the RP column and, subsequently, elution of polar analytes from the PGC column and separation on a hydrophilic interaction liquid chromatography (HILIC) column. To enable separation of polar analytes dissolved in an aqueous matrix by HILIC, the water plug that is flushed from the PGC column is diluted by dosing organic solvent directly upstream of the HILIC column. The developed method was tested by applying target analysis and non-target screening, highlighting the advantage to effectively separate and detect both polar and nonpolar compounds in a single chromatographic run. In the target analysis, the analytes, with a logD at pH 3 ranging from -2.8 to + 4.5, could be enriched and separated. Besides the 965 features in the RP phase, 572 features from real wastewater were observed in the HILIC phase which would otherwise elute in the void time in conventional one-dimensional RP methods.


The (not completely irreversible) population of a misfolded state of cytochrome c under folding conditions.
This paper reports the discovery of a (meta)stable partially unfolded state of horse heart ferricytochrome c that was obtained after exposing the protein to a solution with an alkaline pH of 11.5 for 1 week. Thereafter, the protein did not undergo any detectable change in its secondary and tertiary structure upon adjusting the solution to folding promoting conditions at neutral pH. Spectroscopic data suggest that the misfolded protein exhibits a hexacoordinated low-spin state with a hydroxyl ion as the likely ligand. Below pH 6, a new ligation state emerges with the spectroscopic characteristics of a pentacoordinated quantum mixed state of the heme iron. Gel electrophoresis revealed substantial formation of soluble dimers and trimers at submillimolar concentrations, whereas monomers were dominant at lower, micromolar concentrations. Ultraviolet circular dichroism spectra indicate that oxidized monomers are pre-molten globule to globule-like with a substantial fraction of secondary (helical) structure reminiscent of alkaline state V. The oligomers contain even more helical structure, which suggests domain swapping as the underlying mechanism of their formation. A substantial fraction of the submillimolar mixture of monomers and oligomers underwent a reduction of the heme iron. Its dependence on pH suggests the coupling to a proton transfer process. Altogether, our data indicate a partially unfolded ferricytochrome c conformation with spectroscopic characteristics reminiscent of the recently discovered alkaline isomer V(b), which is stabilized under folding conditions by exposing the protein to a very alkaline pH for an extended period of time.


Complexes of Gilman reagents with C-S and C-N double bonds: sigma or pi bonding?
Upon rapid injection, a variety of thiocarbonyl compounds react with the Gilman reagent Me(2)CuLi at -100 degrees C inside the probe of an NMR spectrometer to give high yields of complexes. Typical examples of substrates include carbon disulfide, methyl dithioacetate, methyl dithiobenzoate, thiobenzophenone, ethylene trithiocarbonate, and phenyl isothiocyanate. Evidence suggesting the formal oxidation state of copper in these complexes to be Cu(III) is presented. The last example was particularly interesting, since it involved a transient intermediate that was identified as a complex with a C-N double bond. Methyl isothiocyanate gave a stable C-N double-bond complex.


[Front roads of Professor M. K. Kuzmin (the 70th anniversary of the Battle for Moscow].
The article is a biographical sketch about the veteran and invalid of Great Patriotic War, the holder of many military decorations, honorary citizen of several cities in the battles for which he was involved, a prominent historian of medicine, doctor of medical sciences, professor Mikhail Kuzmin (1920-2010). Over the years of scientific and teaching life professor M. K. Kuzmin published over 300 scientific publications, including 9 monographs, textbook of the history of medicine, published lectures, and manuals.


Structural insights into the catalytic mechanism of Escherichia coli selenophosphate synthetase.
Selenophosphate synthetase (SPS) catalyzes the synthesis of selenophosphate, the selenium donor for the biosynthesis of selenocysteine and 2-selenouridine residues in seleno-tRNA. Selenocysteine, known as the 21st amino acid, is then incorporated into proteins during translation to form selenoproteins which serve a variety of cellular processes. SPS activity is dependent on both Mg(2+) and K(+) and uses ATP, selenide, and water to catalyze the formation of AMP, orthophosphate, and selenophosphate. In this reaction, the gamma phosphate of ATP is transferred to the selenide to form selenophosphate, while ADP is hydrolyzed to form orthophosphate and AMP. Most of what is known about the function of SPS has derived from studies investigating Escherichia coli SPS (EcSPS) as a model system. Here we report the crystal structure of the C17S mutant of SPS from E. coli (EcSPS(C17S)) in apo form (without ATP bound). EcSPS(C17S) crystallizes as a homodimer, which was further characterized by analytical ultracentrifugation experiments. The glycine-rich N-terminal region (residues 1 through 47) was found in the open conformation and was mostly ordered in both structures, with a magnesium cofactor bound at the active site of each monomer involving conserved aspartate residues. Mutating these conserved residues (D51, D68, D91, and D227) along with N87, also found at the active site, to alanine completely abolished AMP production in our activity assays, highlighting their essential role for catalysis in EcSPS. Based on the structural and biochemical analysis of EcSPS reported here and using information obtained from similar studies done with SPS orthologs from Aquifex aeolicus and humans, we propose a catalytic mechanism for EcSPS-mediated selenophosphate synthesis.


The impact of diuretic therapy on reported sexual function.
--Thiazide diuretics are the most commonly prescribed antihypertensive agent in the treatment of hypertension. No studies to date have isolated conclusively the effect of diuretic therapy on patient quality of life, and particularly on sexual dysfunction. --We carried out a randomized placebo-controlled trial of the effect of thiazide diuretics on quality of life in mildly hypertensive men between the ages of 35 and 70 years. After a 1-month washout period off diuretic therapy, the men were randomized to a 2-month trial of a thiazide treatment or placebo. At the end of the trial, 176 men completed a confidential self-administered questionnaire assessing aspects of social performance, general well-being, and emotional and physiologic states. --Patients taking diuretics reported significantly greater sexual dysfunction than control subjects, including decreased libido, difficulty in gaining and maintaining an erection, and difficulty with ejaculation. Multivariate analysis suggested that the findings were not mediated by low serum potassium levels or by low blood pressure. Sexual dysfunction was noted more often in the confidential self-report questionnaire than in response to direct questions from clinicians. We found no differences between diuretic and placebo groups in measures of mood, general sense of well-being, ability to work, alertness, and social/recreational activities. --Thiazide diuretic use is associated with male sexual dysfunction but does not appear to adversely affect other aspects of quality of life.


Cumulative Exposure to Ideal Cardiovascular Health and Incident Diabetes in a Chinese Population: The Kailuan Study.
It is unclear whether ideal cardiovascular health (CVH), and particularly cumulative exposure to ideal CVH (cumCVH), is associated with incident diabetes. We aimed to fill this research gap. The Kailuan Study is a prospective cohort of 101 510 adults aged 18 to 98 years recruited in 2006-2007 and who were subsequently followed up at 2- (Exam 2), 4- (Exam 3), and 6 (Exam 4)-year intervals after baseline. The main analysis is restricted to those individuals with complete follow-up at all 4 examinations and who had no history of diabetes until Exam 3. Cumulative exposure to ideal CVH (cumCVH) was calculated as the summed CVH score for each examination multiplied by the time between the 2 examinations (score×year). Logistic regression models were used to assess the association between cumCVH and incident diabetes. In fully adjusted models, compared with the lowest quintile of cumCVH, individuals in the highest quintile had ~68% (95% confidence interval [CI] 60-75) lower risk for incident diabetes (compared with 61% [95% CI 52-69] lower risk when using baseline CVH). Every additional year lived with a 1-unit increase in ideal CVH was associated with a 24% (95% CI 21-28) reduction in incident diabetes. Ideal CVH is associated with a reduced incidence of diabetes, but the association is likely to be underestimated if baseline measures of CVH exposure are used. Measures of cumulative exposure to ideal CVH are more likely to reflect lifetime risk of diabetes and possibly other health outcomes. URL: https://www.chictr.org. Unique identifier: ChiCTRTNC-11001489.