Dose, Content, and Context of Usual Care in Stroke Upper Limb Motor Interventions: A Systematic Review.
The objectives of this systematic review were to describe the current dose and content of usual care upper limb motor intervention for inpatients following stroke and examine if context factors alter dose and content. A systematic search (EMBASE, MEDLINE) was completed from January 2015 to February 2023 (PROSPERO CRD42021281986). Studies were eligible if they reported non-protocolised usual care upper limb motor intervention dose data for stroke inpatients. Studies were rated using the Johanna Briggs Institute critical appraisal tool. Data were descriptively reported for dose dimensions of time (on task or, in therapy) and intensity (repetitions, repetition/minute), content (intervention type/mode), and context (e.g., severity strata). Eight studies were included from four countries, largely reflecting inpatient rehabilitation. Time in therapy ranged from 23 to 121 min/day. Time on task ranged from 8 to 44 min/day. Repetitions ranged from 36 to 57/session, and 15 to 282/day. Time on task was lowest in the stratum of people with severe upper limb impairment (8 min/day), the upper limit for this stratum was 41.5 min/day. There was minimal reporting of usual care content across all studies. Upper limb motor intervention dose appears to be increasing in usual care compared to prior reports (e.g., average 21 min/day and 23 to 32 repetitions/session). Context variability suggests that doses are lowest in the stratum of patients with a severely impaired upper limb. Consistent reporting of the multiple dimensions of dose and content is necessary to better understand usual care offered during inpatient rehabilitation.


What is the significance of a positive Propionibacterium acnes culture around a joint replacement?
The purpose of this study was to show the significance of a positive Propionibacterium acnes sample around a joint replacement. Records from the microbiology laboratory data over a 3-year period were reviewed to identify patients with prosthetic joints from whom Propionibacterium acnes was isolated at least once. The medical records of all those patients were retrieved and the demographic, clinical, microbiological and haematological data were collected and examined. The preoperative values of erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) were recorded. Fifty patients underwent a routine revision of a joint arthroplasty; six patients had a joint aspiration. Only one patient had further revision surgery for infection. The preoperative values of ESR and CRP were very variable. The presence of a positive sample around a joint arthroplasty is of uncertain significance. Further studies are needed in order to establish uniform criteria for the diagnosis of infection caused by Propionibacterium acnes.


Encepalomyocarditis virus-induced apoptosis and ultrastructural changes in the lacrimal and parotid glands of mice.
Development of acinar cell apoptosis and ultrastructural changes in the exorbital lacrimal and parotid glands was examined in DBA/2 mice infected with 10(2) PFU/mouse of EMC-D virus. Pyknotic acinar cells, most of which were positive for TUNEL and cleaved caspase-3 and had ultrastructural characteristics of apoptotic cells, developed earlier and were more frequently observed in the parotid gland than in the exorbital lacrimal gland, while the total damage of acinar cells and interstitial infiltration of macrophages were more prominent in the latter than in the former. These findings indicate that EMC-D virus induces acinar cell apoptosis in these glands. In addition, corresponding to the results of the detection of viral RNA signals by in situ hybridization, small aggregates of virus-like particles having typical size and structure of EMC virus were frequently observed in both the cytoplasm and the nucleus of acinar cells in the exorbital lacrimal gland, while they were found only in the cytoplasm of a few acinar cells in the parotid gland. In conclusion, between the exorbital lacrimal and parotid glands, there was a reverse relationship observed between the development of acinar cell apoptosis and that of total damage of acinar cells.


Jun activation domain-binding protein 1 is required for mitotic checkpoint activation via its involvement in hyperphosphorylation of 53BP1.
p53-binding protein 1 (53BP1), a participant in the DNA damage response pathway, has also been implicated in the cellular response to mitotic stress conditions. Here, we sought to broaden our understanding of the protein network surrounding 53BP1 by identifying and characterizing a 53BP1-interacting protein. Yeast two-hybrid screening was performed to identify possible binding partners of 53BP1. To investigate the functional meaning of the interaction, knock-down cells were established by introduction of antisense construct or siRNA into HeLa cells. The hyperphosphorylation of 53BP1 after treatment with nocodazole, a microtubule-interfering agent, was monitored by immunoblotting. And the cell cycle arrest at mitotic phase was measured by flow cytometry after staining with phospho-(Ser10)-histone H3 antibody. Jun activation domain-binding protein 1 (Jab1) was identified as a 53BP1-binding protein, and the interaction between them was confirmed to occur in mammalian cells. We found that nocodazole-induced 53BP1 hyperphosphorylation was abolished in Jab1 knock-down cells. In addition, ectopic overexpression of Jab1 augmented 53BP1 hyperphosphorylation. On the cellular level, Jab1 knock-down cells exhibited reduced mitotic arrest upon exposure to nocodazole, resulting in cellular resistance to the drug. Taken together, these results suggest that Jab1 is required for the hyperphosphorylation of 53BP1 upon mitotic stress conditions and is involved in proper activation of mitotic checkpoint mechanism. Our study also suggests the possibility that modulation of Jab1 activity may be an intriguing approach for enhancing the efficacy of microtubule-interfering anticancer drugs.


Reactions of oxobis(acetylacetonato)-vanadium (IV) with some purine and pyrimidine bases and nucleosides.
A study of the reactions of purine and pyrimidine bases has been initiated with the V(IV) complex [V(O)(acac)2]. Two distinct modes of reaction have been recognized depending on the nature of the base. Simple adduct formation occurs when the base contains a sufficiently basic lone pair, but when bases contain acidic protons bound to nitrogen (inosine, thymine), cleavage of one acetylacetonate ligand occurs to give potentially polymeric species. This behavior is in contrast to that of the inert [Co(NO2)2(acac)2]-ion.


Differential chemokine activation of CC chemokine receptor 1-regulated pathways: ligand selective activation of Galpha 14-coupled pathways.
Chemokines regulate the chemotaxis, development, and differentiation of many cell types enabling the regulation of routine immunosurveillance and immunological adaptation. CC chemokine receptor 1 (CCR1) is the target of 11 chemokines. This promiscuity of receptor-ligand interactions and the potential for functional redundancy has led us to investigate the selective activation of CCR1-coupled pathways by known CCR1 agonists. Chemokines leukotactin-1, macrophage inflammatory protein (MIP)-1alpha, monocyte chemotactic peptide (MCP)-3, RANTES, and MIP-1delta all inhibited adenylyl cyclase activity in cells transiently transfected with CCR1. In contrast, only MIP-1delta was unable to signal via G14-, G16- or chimeric 16z44-coupled pathways. In a stable cell line expressing CCR1 and Galpha14, all of these five chemokines along with hemofiltrate CC chemokine (HCC)-1 and myeloid progenitor inhibitory factor (MPIF)-1 were able to stimulate G(i/o)-coupled pathways, but MIP-1delta, HCC-1 and MPIF-1 were unable to activate G14-mediated stimulation of phospholipase Cbeta activity. In addition, MIP-1delta was unable to promote the phosphorylation of extracellular signal-regulated kinase and c-Jun N-terminal kinase. This suggests that different chemokines are able to selectively activate CCR1-coupled pathways, probably because of different intrinsic ligand efficacies. CCR1 and Galpha14 or Galpha16 are co-expressed in several cell types and we hypothesize that selective activation of chemokine receptors provides a mechanism by which chemokines are able to fine-tune intracellular signaling pathways.


RNA trans-splicing for genodermatoses.
Spliceosome-mediated RNA trans-splicing (SMaRT) is a tool that facilitates the recombination of two distinct pre-mRNA molecules. Its application for gene therapeutic purposes has been hindered by laborious procedures to identify gene-specific molecules. We have established a screening method for the identification of highly functional RNA trans-splicing molecules based on fluorescence reporters, facilitating the generation of most potent therapeutic molecules for the correction of any gene of interest.


Cardiovascular abnormalities and hypertension in the elderly dialysis patient.
Elderly individuals comprise an ever-increasing proportion of patients on renal replacement therapy (RRT). As in younger patients, cardiovascular events are the major cause of death in the elderly on RRT, although the relative proportion succumbing to cardiovascular events is similar to that in young patients. An important observation is that the elderly are particularly likely to experience higher mortality if dialysis technique is suboptimal; they also experience particular benefit from successful renal transplantation. The latter observation has led to an "old for old" program in Europe.


The effect of middle turbinate resection on topical drug distribution into the paranasal sinuses.
During sinus surgery, partial or complete resection of the middle turbinate (MT) is sometimes necessary because of polypoid changes or demineralization. Topical drug delivery to the paranasal sinuses is an integral component in managing chronic rhinosinusitis (CRS) with and without nasal polyposis. The purpose of this study was to examine the role of the MT and MT resection in topical drug distribution into the sinuses via nebulization. We report on a novel technique of quantitatively evaluating the delivery of nebulized dye in cadavers. Endoscopic sinus surgery was performed on 5 fresh cadavers. Complementary colored dyes mixed with saline were successively nebulized using the following protocol: (1) fluorescein prior to MT resection; (2) brilliant green following partial MT resection; and (3) methylene blue following total MT resection. Photodocumentation of the sinuses was performed following each nebulization and standardized photoanalysis was performed. Successive nebulizations with fluorescein, brilliant green, and methylene blue produced a complementary staining pattern that provided an easy side-by-side analysis of the extent of mucosal staining. Dye delivery to the frontal and sphenoid sinuses significantly increased following partial resection of the MT (p = 0.013 and p = 0.0027, respectively) and complete resection of the MT (p = 0.027 and p = 0.027, respectively). Following complete MTR, dye delivery to the maxillary sinus significantly increased compared to baseline (MT intact) (p = 0.0027). Resection of the MT appears to have a significant effect on nebulized drug delivery into the frontal, maxillary, and sphenoid sinuses. Future prospective studies will help delineate the clinical relevance of this effect.


A case-control study of the joint effect of reproductive factors and radiation treatment for first breast cancer and risk of contralateral breast cancer in the WECARE study.
To examined the impact of reproductive factors on the relationship between radiation treatment (RT) for a first breast cancer and risk of contralateral breast cancer (CBC). The Women's Environmental Cancer and Radiation Epidemiology (WECARE) Study is a multi-center, population-based case-control study where cases are women with asynchronous CBC (N = 1521) and controls are women with unilateral breast cancer (N = 2211). Rate ratios (RR) and 95% confidence intervals (CI) were estimated using conditional logistic regression to assess the independent and joint effects of RT (ever/never and location-specific stray radiation dose to the contralateral breast [0, >0-<1Gy, ≥1Gy]) and reproductive factors (e.g., parity). Nulliparous women treated with RT (≥1Gy dose) were at increased risk of CBC compared with nulliparous women not treated with RT, although this relationship did not reach statistical significance (RR = 1.34, 95% CI 0.87, 2.07). Women treated with RT who had an interval pregnancy (i.e., pregnancy after first diagnosis and before second diagnosis [in cases]/reference date [in controls]) had an increased risk of CBC compared with those who had an interval pregnancy with no RT (RR = 4.60, 95% CI 1.16, 18.28). This was most apparent for women with higher radiation doses to the contralateral breast. Among young female survivors of breast cancer, we found some evidence suggesting that having an interval pregnancy could increase a woman's risk of CBC following RT for a first breast cancer. While sampling variability precludes strong interpretations, these findings suggest a role for pregnancy and hormonal factors in radiation-associated CBC.


The Moral Significance of Abortion Inconsistency Arguments.
Most opponents of abortion (OA) believe fetuses matter. Critics argue that OA act inconsistently with regards to fetal life, seeking to restrict access to induced abortion, but largely ignoring spontaneous abortion and the creation of surplus embryos by IVF. Nicholas Colgrove, Bruce Blackshaw, and Daniel Rodger call such arguments inconsistency arguments and contend they do not matter. They present three objections to these arguments - the other beliefs, other actions, and hypocrisy objection. Previously, I argued these objections fail and threaten to undermine ethical inquiry. Colgrove et al. have recently replied, but here, I argue their reply fails as well and raises a new criticism of the other actions' objection. This essay sets out to show, as well as any philosophical argument can, that inconsistency arguments are morally significant.


Experimental studies on effects of temperature on oil and water relative permeability in heavy-oil reservoirs.
A heavy-oil sample derived from a block of Venezuelan oil was used to investigate effects of temperature on relative permeability to oil and water. Measurements of relative permeability were based on one-dimensional core-flow simulated systems using an unsteady-state technique at different temperatures, and then impact rules of temperature dependency were discussed. Both water and heavy oil in cores were reconfigured under the consideration of actual reservoir conditions. Study results suggest that relative permeability is high to oil phase and is very low to water phase, and fluid flow capability is extremely imbalanced between oil and water. As temperature increases, irreducible water saturation linearly increases, residual oil saturation performs a nonlinear decrease, and water saturation exhibits a nonlinear increase at equal-permeability points. The water-wettability of rocks is heightened and overall relative permeability curves shift to the right with increasing temperature; furthermore, two-phase flow area becomes wider and both oil and water relative permeability increases apparently, but the increase ratio of water is less than that of oil.


Association of body mass index and weight change with death in patients with advanced cancer.
Conventional diagnostic criteria for cachexia are based on Western studies. The aim of this study was to investigate trends in body mass index (BMI) and the rate of weight change associated with adverse outcomes in Asian patients with advanced cancer. This retrospective cohort study was conducted using the administrative claims database compiled by the Japan Medical Data Center Inc. The study was conducted between April 2014 and September 2022 on patients with advanced cancer. A Cox regression model was used to perform a restricted cubic spline analysis with four knots for BMI and weight change. Additionally, thin-plate smoothed splines were used to generate contour plots of the odds ratios of BMI and weight change for mortality. The study analyzed 48 600 patients. The mean age was 71.9 ± 10 y. There were 33 051 men (68%) and 17 853 deaths (37%). The smoothed splines for BMI showed that low BMI was associated with high hazard ratio (HR, [95% confidence interval]; BMI = 18.5 kg/m2, HR, 1.2; [1.1-1.2]; BMI = 25 kg/m2, HR, 0.9; [0.9-0.9]). The smoothed spline of weight change showed a higher HR with an increasing rate of weight change (weight change rate -10%, HR, 1.1; [1.1-1.2]; weight change rate +10%, HR, 1; [1-1]). In the thin-plate smoothed spline analysis, patients with BMI <17 to 18.5 kg/m2 were at a higher risk for death regardless of weight change. Low BMI, a large degree of weight change, or a combination of both, predicted death in Asian patients with advanced cancer.


A new species and three new records of Chloroperlidae (Plecoptera) from northeastern China.
A new species of the genus Suwallia Ricker, 1943, S. kuandian sp. nov., from Liaoning Province of northeastern China is described, illustrated, and compared with similar species. Suwallia asiatica Zhiltzova Levanidova, 1978 is reported for the first time from China. Additionally, Sweltsa illiesi Zhiltzova Levanidova, 1978 and the chloroperlid genus Alaskaperla Stewart DeWalt, 1991 are reported for the first time from China. The habitus and diagnostic features of these species are illustrated with color images.


Putting people at heart of nursing.
A master's degree to introduce nurses to the concept of person-centred practice has been launched by Queen Margaret University.


Extended application of flow microfluorometry by means of dual laser excitation.
A dual laser beam excitation device for flow analysis of biological particles has been developed. The aid of this arrangement is to increase the range of fluorescent agents employed so far in quantitative and qualitative cytochemistry. Combining an argon ion and a helium-cadmium laser two color fluorescence measurements were performed employing propidium iodide as a DNA stain and fluorescamine which stains total protein in fixed cells. Energy transfer processes between the antibiotic and DNA specific dye mithramycin and propidium iodide both being bound to nuclear chromatin were analyzed. Utilization of energy transfer processes is generally discussed as a mean to extract information about the structure and conformation of nuclear chromatin in situ. The application of a crypton ion laser with three lines near 400 nm and a single line at 350 nm having a light output in each range of nearly one Watt gives the opportunity of utilizing DNA fluorochromes which have an excitation maximum in the deep blue region, DNA spectra are shown employing mithramycin, the benzimidazol derivative 33258 (Hoechst) and the indol compound DAPI which has a high DNA specificity combined with a great stability under UV illumination. By separating two focussed laser beams at their intersecting points with the liquid sample stream the trajectory of each flowing cell crosses the beams sequentially, which causes a solitary dual excitation of each cell. The advantages of a solitary excitation device compared with a simultaneous one is discussed.


Ischemic preconditioning or p38 MAP kinase inhibition attenuates myocardial TNF alpha production and mitochondria damage in brief myocardial ischemia.
Coronary artery occlusion increased the TNF alpha level in the membrane fraction of the rat heart, almost maximally at 30 min. TNF alpha immunofluorescence labeled streak-like reticular structures inside of the cardiomyocyte but not vascular or interstitial cells in myocardial ischemia. Immuno-electron microscopy confirmed the localization of TNF alpha between myofibrils, mitochondria, or other membrane structures in the ischemic cardiomyocyte. Ischemic preconditioning (IP) is the protection of myocardium conferred by cycles of brief ischemia-reperfusion. The increases in TNF alpha production, as well as phosphorylation of p38 MAP kinase and S6 kinase after ischemia were inhibited by IP or p38 MAP kinase inhibitors (SB203580, FR167653). TNF alpha production appeared to be regulated possibly at the post-transcriptional step by ribosomal S6 phosphorylation given that IP did not suppress TNF alpha mRNA up-regulation and was independent of NFkappaB activation. Electron microscopy (EM) showed mitochondria damage in ischemic cardiomyocyte, which was inhibited either by IP or SB203580. This is the first demonstration of the TNF alpha up-regulation in membrane structures of ischemic cardiomyocyte through p38 MAP kinase-mediated post-transcriptional mechanism, in association with mitochondrial damage.


Monitoring Photochemical Reactions Using Marangoni Flows.
We evaluated the sensitivity and time resolution of a technique for photochemical reaction monitoring based on the interferometric detection of the deformation of liquid films. The reaction products change the local surface tension and induce Marangoni flow in the liquid film. As a model system, we consider the irradiation of the aliphatic hydrocarbon squalane with broadband deep-UV light. We developed a numerical model that quantitatively reproduces the flow patterns observed in the experiments. Moreover, we present self-similarity solutions that elucidate the mechanisms governing different stages of the dynamics and their parametric dependence. Surface tension changes as small as Δγ = 10-6 N/m can be detected, and time resolutions of <1 s can be achieved.


Supraphysiological hCG day estradiol levels can predict pregnancy-associated plasma protein A levels in maternal serum in the first trimester.
To investigate the effect of hCG day estradiol (hCG-E2) used in Down Syndrome screening on maternal serum levels of PAPP-A in fresh in vitro fertilization (IVF) cycles. This study was a retrospective analysis of a cohort that resulted in a single pregnancy after a total of 92 fresh IVF cycles. The primary outcome of this study was to determine the effect of fresh IVF cycle parameters on the PAPP-A level and the cutoff value for hCG-E2 predicting a low PAPP-A level, while the secondary outcome was to determine whether the effect of IVF parameters on the PAPP-A level was significant. There was a negative correlation between PAPP-A levels and the number of hCG-E2 and grade 1 embryos (respectively, p = .049; .047), while a positive correlation was observed between baby weight at birth and the PAPP-A (p < .05). At a PAPP-A value of 0.82, the difference between the two groups, in terms of hCG-E2, the number of grade 1 embryos, and pregnancy-related complications was significant (p = .050; .029; .033, respectively). The threshold value of hCG-E2 affecting PAPP-A levels was statistically significant (AUC = 0.618; p = .050; hCG-E2 = 4869.5 pg/ml). In the model, an increase in the number of grade 1 embryos resulted in higher PAPP-A levels (OR = 2.26; p = .044). The fact that the hCG-E2 cutoff value, which lowers PAPP-A, reflects excessive ovarian stimulation argues for the correction of the dual screening test in a subset of patients with high response to the first-trimester screening test.


Tissue-plasminogen activator stimulates endothelial cell migration in wound assays.
The ability of tissue plasminogen activator (tPA) to induce human umbilical vein endothelial (HUVE) cell migration was studied using an in vitro, serum-free wound assay system. At pharmacological doses, tPA stimulated HUVE cell migration dose-dependently. Treatment of cells with epsilon amino caproic acid (EACA) to detach cell-surface and extracellular matrix bound plasminogen, which could lead to plasmin generation, resulted in increased HUVEcell migration on stimulation with tPA.Plasminogen activator inhibitor-1 (PAI-1), a natural plasminogen activator inhibitor, abolished tPA-induced HUVEcell migration. These results demonstrate for the first time that tPA is capable of stimulating endothelial cell migration in wound assays and this effect is susceptible to PAI-1 inhibition.


UDP-glucose 4-epimerase from Saccharomyces fragilis. Involvement of sulfhydryl group(s) at the active site.
UDPglucose-4-epimerase (EC 184.108.40.206) from Saccharomyces fragilis is inactivated by 0.1 mM 5,5'-dithiobis-(2-nitrobenzoate) in 6 min. Unlike p-chloromercuribenzoate-inactivated or heat-inactivated enzymes, the dithiobisnitrobenzoate-inactivated enzyme retains the dimeric structure and NAD is not dissociated from the protein moiety. Inactivation of the enzyme by dithiobisnitrobenzoate can not therefore be attributed to any subsequent loss of structural integrity or to the detachment of the cofactor from the apoenzyme. The inactivated enzyme can be almost fully reactivated in the presence of mercaptoethanol and characteristic properties of native enzyme are regained. The inactivation by dithiobisnitrobenzoate can be substantially protected by UDPglucose or UDPgalactose indicating a possible critical involvement of one or more sulfhydryl groups at the active site.


Two types of high-threshold calcium currents inhibited by omega-conotoxin in nerve terminals of rat neurohypophysis.
1. The neurohypophysis comprises the nerve terminals of hypothalamic neurosecretory cells, which contain arginine vasopressin (AVP) and oxytocin. The secretory terminals of rat neurohypophyses were acutely dissociated. The macroscopic calcium currents (ICa) of these isolated peptidergic terminals were studied using 'whole-cell' patch-clamp recording techniques. 2. There are two types ('Nt' (where the subscript 't' denotes terminal) and 'L') of high-threshold voltage-activated ICa in the terminals, which can be distinguished by holding at different potentials i.e. -90 and -50 mV. Replacement of Ca2+ in the bathing solution by Ba2+ increased the amplitude of ICa, primarily due to an increase in the L-type component. Both inward currents were eliminated by adding 50 microM-Cd2+ or when in a Ca(2+)-free bathing solution. 3. omega-Conotoxin GVIA (omega-CgTx) has been widely used as a Ca2+ channel blocker. However, whether this toxin can discriminate between different types of Ca2+ channels is still a subject of controversy. We applied omega-CgTx over a wide range of concentrations (0.01-2 microM) to examine its effects on both Nt- and L-type ICa in these terminals. At a concentration of 30 nM, omega-CgTx selectively reduced, by 48%, the amplitude of Nt-type ICa. In contrast, a higher concentration (300 nM) of omega-CgTx was necessary to inhibit the L-type ICa. 4. omega-CgTx inhibited both Nt- and L-type ICa in a dose-dependent manner, and the half-maximum inhibition (IC50) of the ICa by the toxin was 50 and 513 nM, respectively, which was approximately a tenfold difference. The reduction in both types of currents did not result from any shift in their current-voltage or steady-state inactivation relationships. 5. In contrast, omega-CgTx, at a concentration of 300 nM, had no effect on the tetrodotoxin-sensitive sodium current (INa) of the isolated peptidergic nerve terminals. Furthermore, omega-CgTx did not reduce the long-lasting, non-inactivating ICa in the isolated non-neuronal secretory cells of the pars intermedia (PI) (intermediate lobe of the pituitary). 6. Our studies suggest that omega-CgTx might exert specific blocking effects on both Nt- and L-type Ca2+ channels, but that in the isolated peptidergic nerve terminals, the Nt-type component is more susceptible to this toxin.


'Straight lateral traction' in selected supracondylar fractures of the humerus in children.
A method of treating completely displaced supracondylar fractures of the humerus in children by 'straight lateral traction', as originally used by John Dunlop, is recommended. The results in 26 children, selected because manipulation had failed to achieve reduction, is examined, and indicates that this treatment is satisfactory for some problem fractures, leading usually to good results in terms of resolution of complications and return of function. It also avoids the ugly deformities of the elbow which follow reliance on a single form of treatment for all supracondylar fractures. Failure of anatomical reduction by the appliance is shown to be in the plane of elbow movement, where normal physiological mechanisms correct the bone by remodelling. Where there is epiphyseal damage, deformities may occur with any form of treatment.


Swab-Seq: A high-throughput platform for massively scaled up SARS-CoV-2 testing.
The rapid spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is due to the high rates of transmission by individuals who are asymptomatic at the time of transmission1,2. Frequent, widespread testing of the asymptomatic population for SARS-CoV-2 is essential to suppress viral transmission. Despite increases in testing capacity, multiple challenges remain in deploying traditional reverse transcription and quantitative PCR (RT-qPCR) tests at the scale required for population screening of asymptomatic individuals. We have developed SwabSeq, a high-throughput testing platform for SARS-CoV-2 that uses next-generation sequencing as a readout. SwabSeq employs sample-specific molecular barcodes to enable thousands of samples to be combined and simultaneously analyzed for the presence or absence of SARS-CoV-2 in a single run. Importantly, SwabSeq incorporates an in vitro RNA standard that mimics the viral amplicon, but can be distinguished by sequencing. This standard allows for end-point rather than quantitative PCR, improves quantitation, reduces requirements for automation and sample-to-sample normalization, enables purification-free detection, and gives better ability to call true negatives. After setting up SwabSeq in a high-complexity CLIA laboratory, we performed more than 80,000 tests for COVID-19 in less than two months, confirming in a real world setting that SwabSeq inexpensively delivers highly sensitive and specific results at scale, with a turn-around of less than 24 hours. Our clinical laboratory uses SwabSeq to test both nasal and saliva samples without RNA extraction, while maintaining analytical sensitivity comparable to or better than traditional RT-qPCR tests. Moving forward, SwabSeq can rapidly scale up testing to mitigate devastating spread of novel pathogens.


Contaminant-induced oxidative stress in fish: a mechanistic approach.
The presence of reactive oxygen species (ROS) in living organisms was described more than 60 years ago and virtually immediately it was suggested that ROS were involved in various pathological processes and aging. The state when ROS generation exceeds elimination leading to an increased steady-state ROS level has been called "oxidative stress." Although ROS association with many pathological states in animals is well established, the question of ROS responsibility for the development of these states is still open. Fish represent the largest group of vertebrates and they inhabit a broad range of ecosystems where they are subjected to many different aquatic contaminants. In many cases, the deleterious effects of contaminants have been connected to induction of oxidative stress. Therefore, deciphering of molecular mechanisms leading to such contaminant effects and organisms' response may let prevent or minimize deleterious impacts of oxidative stress. This review describes general aspects of ROS homeostasis, in particular highlighting its basic aspects, modification of cellular constituents, operation of defense systems and ROS-based signaling with an emphasis on fish systems. A brief introduction to oxidative stress theory is accompanied by the description of a recently developed classification system for oxidative stress based on its intensity and time course. Specific information on contaminant-induced oxidative stress in fish is covered in sections devoted to such pollutants as metal ions (particularly iron, copper, chromium, mercury, arsenic, nickel, etc.), pesticides (insecticides, herbicides, and fungicides) and oil with accompanying pollutants. In the last section, certain problems and perspectives in studies of oxidative stress in fish are described.


Whipple's disease presenting as weight gain and constipation in a Chinese woman.
Whipple's disease is a chronic infection due to Tropheryma whipplei, commonly reported in the Caucasian but not in the Chinese population. A 52-year-old female with good past health, was diagnosed with Whipple's disease, presenting with constipation, unintentional weight gain, and fleeting polyarthralgia. Investigations prior to admission showed raised CA125 and computed tomography of the abdomen showed multiple retroperitoneal mesenteric lymphadenopathies. Extensive investigations performed on secondary causes of weight gain were unrevealing. Subsequent PET-CT scan revealed generalized lymphadenopathy involving the left deep cervical, supraclavicular, and retroperitoneal mesenteric area. Excisional biopsy of the left supraclavicular lymph node was performed, with histology showing infiltrations of Periodic acid-Schiff positive foamy macrophages. T. whipplei DNA was detected in her serum, saliva, stool, and lymph node by PCR targeting the 16S ribosomal RNA gene. She was started on intravenous ceftriaxone, and then stepped down to oral antibiotics for a total of 44 months. The recurrence of fever after 12 days of ceftriaxone raised the suspicion of Immune Reconstitution Inflammatory Syndrome (IRIS). Serial imaging showed a gradual reduction in the size of retroperitoneal lymphadenopathies. Literature review on Whipple's disease in the Chinese population identified 13 reports of detectable T. whipplei DNA in clinical specimens. The majority of the cases were pneumonia, followed by culture-negative endocarditis, encephalitis, and skin and soft tissue infection. However, most patients with pneumonia were diagnosed based on next generation sequencing alone, with the resolution of pulmonary infiltrates without adequate duration of antibiotics, suggesting the possibility of colonization instead of infection. The recommendation of long-term doxycycline suppression after treatment may be supported by the slow response of retroperitoneal lymphadenopathies to antibiotics in our patient. Unintentional weight gain and constipation could be atypical presentations of Whipple's disease. It is a rare disease in the Chinese population despite the advancement of molecular techniques in the diagnosis of infections. A prolonged course of antibiotics may be required due to slow clinical response as documented by serial imaging in our case. The possibility of IRIS should be considered in patients with breakthrough fever during treatment of Whipple's disease.


In vitro exploration of potential mechanisms of toxicity of the human hepatotoxic drug fenclozic acid.
The carboxylic acid NSAID fenclozic acid exhibited an excellent preclinical safety profile and promising clinical efficacy, yet was withdrawn from clinical development in 1971 due to hepatotoxicity observed in clinical trials. A variety of modern in vitro approaches have been used to explore potential underlying mechanisms. Covalent binding studies were undertaken with [(14)C]-fenclozic acid to investigate the possible role of reactive metabolites. Time-dependent covalent binding to protein was observed in NADPH-supplemented liver microsomes, although no metabolites were detected in these incubations or in reactive metabolite trapping experiments. In human hepatocytes, covalent binding was observed at lower levels than in microsomes and a minor uncharacterizable metabolite was also observed. In addition, covalent binding was observed in incubations undertaken with dog and rat hepatocytes, where a taurine conjugate of the drug was detected. Although an acyl glucuronide metabolite was detected when liver microsomes from human, rat and dog were supplemented with UDPGA, there was no detectable UDPGA-dependent covalent binding. No effects were observed when fenclozic acid was assessed for P450-dependent and P450-independent cytotoxicity to THLE cell lines, time-dependent inhibition of five major human cytochrome P450 enzymes, inhibition of the biliary efflux transporters BSEP and MRP2 or mitochondrial toxicity to THLE or HepG2 cells. These data suggest that Phase 1 bioactivation plays a role in the hepatotoxicity of fenclozic acid and highlight the unique insight into mechanisms of human drug toxicity that can be provided by investigations of biotransformation and covalent binding to proteins.


Hybrid nanofibrous yarns based on N-carboxyethylchitosan and silver nanoparticles with antibacterial activity prepared by self-bundling electrospinning.
Hybrid nanofibrous materials with antibacterial activity consisting of yarns from N-carboxyethylchitosan (CECh) and poly(ethylene oxide) (PEO) that contain 5 wt% or 10 wt% silver nanoparticles (AgNPs) were prepared. This was achieved by electrospinning using formic acid as a solvent and as a reducing agent for silver ions. AgNO₃ was used as an Ag(+)-containing salt. Its concentration was selected to be 0.02 mol/L or 0.04 mol/L in order the content of the AgNPs in the electrospun nanofibers to be 5 wt% or 10 wt%, respectively. The self-bundling of the fibers into yarns with a mean diameter of ca. 35 μm was enabled only by using a grounded needle electrode. The reduction of the silver ions to an elemental silver was evidenced by UV-vis spectroscopy, X-ray diffraction (XRD), and X-ray photoelectron spectroscopy (XPS). The transmission electron microscopy (TEM) analyses revealed that AgNPs formed at AgNO₃ concentration of 0.02 mol/L were with a mean diameter of 4±0.5 nm and were distributed uniformly within the fiber. The increase of AgNO₃ concentration to 0.04 mol/L led to the preparation of AgNPs with a higher mean diameter and a broader diameter distribution as well as to aggregate formation. The performed studies on the antibacterial activity of CECh/PEO/AgNPs fibrous materials against Staphylococcus aureus showed that at AgNPs content of 5 wt% the mats had bacteriostatic, and at AgNPs content of 10 wt%-bactericidal activity.


Calcium-sensitive ROS-GC1 signaling outside of photoreceptors: a common theme.
Until now, ROS-GC1 signal transduction system was thought to be exclusive to photoreceptors in the retina. Two recent reports, however, now show that this is not the case. In one, the ROS-GC1 signal transduction system has been identified and characterized in pinealocyte neurons. This signaling is modulated by norepinephrine. However, the response of the individual pinealocyte neuron to the norepinephrine signal depends on whether the GCAP1-linked (results in hyperpolarization) or S100beta-linked (results in depolarization) pathway is operational in the pinealocyte. The GCAP1-linked pathway results in hyperpolarization, while the S100beta-linked pathway, in depolarization. The two pathways are mutually exclusive. In the other report, the calcium-modulated ROS-GC1:GCAP1 signaling system has been discovered in mitral cells of the olfactory bulb. These findings raise the possibility that a common theme of calcium-modulated ROS-GC signaling may be utilized in a wide variety of neurosensory cells. This idea is also supported from evolutionary and functional perspectives.


Mapping the spatial variability of subsurface resistivity by using vertical electrical sounding data and geostatistical analysis at Borena Area, Ethiopia.
Vertical electrical sounding survey has been done to map and visualize resistivity distribution at Borena basin. The area is situated in Southern Main Ethiopian Rift, where Rirriba rift, Mega rift and associated fractures define the structural setting of the area. It is covered by Quaternary deposits, Quaternary and Tertiary basaltic rocks and Precambrian metamorphic rocks. Bullal basaltic formation outstretch in the Rirriba rift and it is thought to be potential groundwater aquifer. About 288 Vertical Electrical Sounding data were collected. Inflection and extreme points were used to identify characteristic points. Variograms are modeled and kriging interpolation is used to map distribution of resistivity, determined from characteristic points, in the area. Very low to low resistivity variations are mapped in northern end of the study area, whereas medium to moderately resistive ground are mapped in the middle and southern part of the area. The low resistivity horizon at the shallow subsurface could be due to salinity since the area occupy numerous saline craters and maars. Approximate mapping of large sets of Vertical Electrical Sounding data with geostatistical treatment has facilitated the interpretation and provided a sound picture of the subsurface. • Inflection and extreme points were extracted from smothed VES curves to identify characteristic points. • Variograms are modeled and kriging interpolation is used to map 3D distribution of resistivity data.


The first case of actinide triple helices: pH-dependent structural evolution and kinetically-controlled transformation of two supramolecular conformational isomers.
The first actinide triple helices, including two supramolecular conformational isomers of uranium(VI), have been synthesized with the aid of a flexible V-shaped ligand and a rigid aromatic base. The isomers exhibit an intriguing pH-dependent structural evolution and a kinetically-controlled transformation via a novel conformational rearrangement of the organic base.


A chronopharmacological diagnostic test and treatment for bipolar disorder and depression: nitric oxide release during sleep causes it to become depressogenic in a subset of patients.
In some patients sleep can have a depressogenic effect and sleep reduction can result in mania. The hypothesis on which this model is based postulates that it is nitric oxide release during sleep that causes it to become depressogenic in a subset of patients. That subset is comprised of patients with genetic polymorphisms in nitric oxide genes and/or in serotonin and melatonin genes. The nature of the dynamic equilibrium between nitric oxide, on the one hand, and serotonin and melatonin, on the other, determines whether or not sleep is likely to have a depressogenic effect. Changes throughout the sleep cycle thereby impart a chronological factor to the dynamic equilibrium. Other writers have postulated, but not defined, a certain so-called "critical period" during the sleep cycle when sleep can become particularly depressogenic. This model assumes that in fact there is such a critical period and that it is stage IV sleep. In addition to the chronological influences on the dynamic equilibrium, the aging process also exerts an influence. The increased sleep fragmentation associated with the aging process causes stage IV sleep to become increasingly more depressogenic because of a concept called delta rebound. Delta rebound results in deeper sleep intensity. It is associated with increased nitric oxide production and the concomitant serotonin suppression that is associated with it. The chronopharmacological test that is proposed challenges the dynamic equilibrium between nitric oxide, on the one hand, and serotonin and melatonin on the other. It consists of melatonin taken at bedtime and the antidepressant moclobemide, taken as close to the onset of stage IV sleep as possible. If there is a noticeable lessening of depressive symptoms the following morning, then drug treatments that increase serotonin and/or melatonin are appropriate and drugs that suppress or block serotonin or melatonin, or that increase nitric oxide, are not.


Can deceased donor with recurrent primary brain tumor donate kidneys for transplantation?
Kidney transplantation from deceased donors is in its infancy in India. Cadaver organ donation was accepted legally in 1994 by the "Human Organs Transplantation Act." Marginal donors are now accepted by many centers for kidney transplantation. We report a case of procurement of both kidneys from a young deceased donor having recurrent primary brain tumor, transplanted into two adult recipients with successful outcome.


Sharing roads with automated vehicles: A questionnaire investigation from drivers', cyclists' and pedestrians' perspectives.
Despite the promised benefits, the introduction of Automated Vehicles (AVs) on roads will be confronted by many challenges, including public readiness to use those vehicles and share the roads with them. The risk profile of road users is a key determinant of their safety on roads. However, the relation of such risk profiles to road users' perception of AVs is less known. This study aims to address the above research gap by conducting a cross-sectional survey to investigate the acceptance of Fully Automated Vehicles (FAVs) among different non-AV-user groups (i.e., pedestrians, cyclists, and conventional vehicle drivers). A total of 1205 road users in Queensland (Australia) took part in the study, comprising 456 pedestrians, 339 cyclists, and 410 drivers. The Theory of Planned Behaviour (TPB) is used as the theoretical model to examine road users' intention towards sharing roads with FAVs. The risk profile of the participants derives from established behavioural scales and individual characteristics are also included in the acceptance model. The study results show that pedestrians reported lowest intention in terms of sharing roads with FAVs among the three groups. Drivers and cyclists in a lower risk profile group were more likely to report higher intention to share roads with FAVs than those in a higher risk profile group. As age increased, pedestrians were less likely to accept sharing roads with FAVs. Drivers who had more exposure time on roads were more likely to accept sharing roads with FAVs. Male drivers reported higher intention towards sharing roads than female drivers. Overall, the study provides new insights into public perceptions of FAVs, specifically from the non-AV-user perspective. It sheds light on the obstacles that future AVs may encounter and the types of road users that AV manufacturers and policymakers should consider closely. Specifically, groups such as older pedestrians and road users who engage in more risky behaviours might resist or delay the integration of AVs.


Transcutaneous PO2, PCO2 and heart rate patterns during normal postnatal adaptation and respiratory distress.
Three groups of neonates were observed during their first 5 days of postnatal life. Healthy term babies (N = 22) did not differ in their heart rate variability (HRV) nor heart rate (HR) from the healthy preterm babies (N = 21) during the first 3 days. On day 5 beat-to-beat variability of term babies was greater. In healthy term and preterm infants HRV increased between days 1 and 3. HRV of the babies with respiratory distress syndrome (RDS) (N = 1) remained low throughout the study period. HR of the babies with RDS was significantly higher compared to the other groups on the days 1 and 3. Transcutaneous PO2 was high in healthy preterm infants compared to the other groups. The correlation between PaCO2 and PtcO2 in RDS was insufficient to make clinical judgement. PtcCO2 of all neonates tended to rise on the day 5 but the rise was most prominent in the infants with RDS. The correlation of PaCO2 and PtcCO2 was good. In RDS high HR, low HRV and elevated PtcCO2 were the typical findings and present throughout the study period.


Urinary excretion of 5-hydroxyindolacetic acid in occupational exposure to polyvinyl chloride and polyurethane foams.
The effect of plastics as polyurethane foams and polyvinyl chloride on the metabolism of tryptophan was investigated by determining the concentration of 5-hydroxyindoleacetic acid (5-HIAA) as its metabolic end product excreted in the urine. Investigation of persons occupationally exposed to polyurethane and polyvinyl chloride showed that these plastics inhibit the metabolism of tryptophan, causing increased excretion of its easily measurable metabolite 5-HIAA. This fact points to the possibility of practical use of this laboratory test which, together with other biological parameters, could improve methods for early detection of occupational exposure to polyurethanes and polyvinyl chloride.


Cystic fibrosis detection in high-risk Egyptian children and CFTR mutation analysis.
Knowledge about Cystic Fibrosis (CF) in Egypt is very limited. The objective of this study was to screen for CF in Egyptian children with suggestive clinical features and to identify causative genetic mutations. Sixty-one patients from the Chest Unit, Cairo University Children's Hospital, Egypt, were included. Subjects presented with persistent or recurrent respiratory symptoms, failure to thrive, diarrhea and/or steatorrhea and unexplained persistent jaundice. Patients were screened using the CF Indicatortrade mark sweat test system (PolyChrome Medical, Inc., Brooklyn Center, MN). A quantitative sweat testing was conducted on 10 of the 12 positive patients. Seven probands and one sibling underwent molecular analysis by direct DNA sequencing of the coding region and of the intronic sequences adjacent to the 27 exons of the CFTR gene. Of 61 patients, 12 (20%) had positive sweat chloride screening. Ten of the 12 patients underwent quantitative sweat testing and were positive. Eight CFTR sequence changes were identified in seven affected probands and two were confirmed in one sibling by direct DNA sequencing. The study results suggest that CF is more common in Egypt than previously anticipated. Larger studies are warranted to identify the incidence, molecular basis and clinical pattern of CF in the Egyptian population.


Positive surgical margins and their locations in specimens are adverse prognosis features after radical cystectomy in non-metastatic carcinoma invading bladder muscle: results from a nationwide case-control study.
To compare the prognoses associated with positive surgical margins (PSMs) according to their urethral, ureteric and/or soft tissue locations in patients with pN0 M0 bladder cancer who have not undergone neoadjuvant chemotherapy. A retrospective, case-control study was conducted between 1991 and 2011 using data from 17 academic centres in France. A total of 154 patients (cases) with PSMs met the eligibility criteria and were matched according to centre, pT stage, gender, age and urinary diversion method with a population-based sample of 154 patients (controls) from 3651 patients who had undergone cystectomies. The median follow-up period was 23.9 months. Multivariable Cox regression analysis was used to test the effects of PSMs on local recurrence (LR)-free survival, metastatic recurrence (MR)-free survival and cancer-specific survival (CSS). The 5-year LR-free survival and CSS rates of patients with urethral and soft tissue PSMs were lower than those in the control group. A significant decrease in CSS was associated with soft tissue PSMs (P = 0.003, odds ratio = 0.425, 95% confidence interval 0.283-0.647). The prognosis was not affected in cases of ureteric PSMs. Soft tissue PSMs were associated with poor CSS rates in patients with pN0 M0 bladder cancer. A correlation between urethrectomy and a reduction of the risk of LR in a urethral PSM setting was observed.


Machining efficiency of Flex-R, K-Flex, Trio-Cut, and S Files.
Many root canal instruments with several different designs have been introduced during recent years. This article evaluates the machining properties of four such instruments: the Flex-R, K-Flex, Trio-Cut, and S file. There are large variations in the machining properties of these instruments. The results are comparable to findings when evaluating conventional root canal files which show that the Hedstrom file is much more efficient when considering removal of material than the K file. The Flex-R file, K-Flex file, and Trio-Cut file are similar in design. The two former brands perform in a similar fashion, while the Trio-Cut file is less efficient than any of the evaluated K files. The S file performs as a Hedstrom file which can be expected from its similarity in design to that instrument.


TIMP-2 mediates the anti-invasive effects of the nitric oxide-releasing prodrug JS-K in breast cancer cells.
Tumor invasion and metastasis remain a major cause of mortality in breast cancer patients. High concentrations of nitric oxide (NO) suppress tumor invasion and metastasis in vivo. NO prodrugs generate large amounts of NO upon metabolism by appropriate intracellular enzymes, and therefore could have potential in the prevention and therapy of metastatic breast cancer. The present study was designed to determine the effects of the NO-releasing prodrug O2-(2,4-dinitrophenyl) 1- [(4-ethoxycarbonyl)piperazin-1-yl]diazen-1-ium-1,2-diolate (JS-K) on breast cancer invasion and the mechanisms involved. MDA-MB-231, MDA-MB-231/F10, and MCF-7/COX-2 were the three breast cancer cell lines tested. NO levels were determined spectrophotometrically using a NO assay kit. Invasion and the expression of matrix metalloproteinases (MMPs) and tissue inhibitor of MMPs were determined using Matrigel invasion assays, an MMP array kit and ELISAs. The activity and expression of extracellular signal-regulated kinase 1/2, p38, and c-Jun N-terminal kinase mitogen-activated protein kinases were determined using western blot analyses. Under conditions by which JS-K was not cytotoxic, JS-K significantly decreased (P < 0.05) the invasiveness of breast cancer cells across the Matrigel basement membrane, which was directly correlated with NO production. JS-43-126, a non-NO-releasing analog of JS-K, had no effect on NO levels or invasion. JS-K increased (P < 0.05) TIMP-2 production, and blocking TIMP-2 activity with a neutralizing antibody significantly increased (P < 0.05) the invasive activity of JS-K-treated cells across Matrigel. JS-K decreased p38 activity, whereas the activity and the expression of extracellular signal-regulated kinase 1/2 and c-Jun N-terminal kinase were unaffected. We report the novel findings that JS-K inhibits breast cancer invasion across the Matrigel basement membrane, and NO production is vital for this activity. Upregulation of TIMP-2 production is one mechanism by which JS-K mediates its anti-invasive effects. JS-K and other NO prodrugs may represent an innovative biological approach in the prevention and treatment of metastatic breast cancer.


Activation of transcription factors by drugs inducing oxidative stress in rat liver.
Chemically induced oxidative stress of the liver associates with gene reprogramming and activation of some transcription factors (TFs), in particular nuclear factor-kappaB (NF-kappaB). We have now investigated other TFs, such as activator protein-1 (AP-1) and hypoxia inducible factor-1 (HIF-1) that we had shown to be activated in rat liver during heat shock, ischemia or post-ischemic reperfusion, and signal transducer and activator of transcription (STAT), CCAAT/enhancer binding protein (C/EBP) and hepatocyte nuclear factor-1 (HNF-1), which may be involved in the response of the liver to injury. The expression of target genes, containing consensus sequences for these TFs was assessed by Northern and Western blot analysis. The rats were treated with buthionine-sulfoximine, nitrofurantoin (NF) or phorone (Ph), which cause liver oxidative stress with different mechanisms. All these agents activated AP-1, known to depend on redox state, HIF-1, initially described as an hypoxia-responsive TF, and STAT3, generally connected to the response to cytokines. HNF-1 a constitutive TF associated with liver-specific gene expression was not affected. The composition of AP-1 was slightly different according to the drug used for treatment. The levels of the mRNAs for heme oxygenase-1 (HO-1), Aldolase A and alpha(1)-acid glycoprotein as well as the corresponding proteins increased after the treatments, thus, indicating that the activation of the TFs was functional. These observations suggest that the treatment of rats with drugs inducing oxidative stress causes a broad spectrum of changes in gene expression with features common to stresses generally considered as separate entities.


The ribosomal A site-bound sense and stop codons are similarly positioned towards the A1823-A1824 dinucleotide of the 18S ribosomal RNA.
Positioning of the mRNA codon towards the 18S ribosomal RNA in the A site of human 80S ribosomes has been studied applying short mRNA analogs containing either the stop codon UAA or the sense codon UCA with a perfluoroaryl azide group at the uridine residue. Bound to the ribosomal A site, a modified codon crosslinks exclusively to the 40S subunits under mild UV irradiation. This result is inconsistent with the hypothesis [Ivanov et al. (2001) RNA 7, 1683-1692] which requires direct contact between the large rRNA and the stop codon of the mRNA as recognition step at translation termination. Both sense and stop codons crosslink to the same A1823/A1824 invariant dinucleotide in helix 44 of 18S rRNA. The data point to the resemblance between the ternary complexes formed at elongation (sense codon.aminoacyl-tRNA.AA dinucleotide of 18S rRNA) and termination (stop codon.eRF1.AA dinucleotide of 18S rRNA) steps of protein synthesis and support the view that eRF1 may be considered as a functional mimic of aminoacyl-tRNA.


Reactive hyperemic responses in systemic sclerosis patients and healthy controls.
Hyperemic responses after arterial occlusion were investigated in patients with Raynaud's phenomenon due to systemic sclerosis (SSc) and in healthy controls. The hyperemia due to arterial occlusion for 2 min was measured by laser-Doppler flowmetry. If the hyperemic response was absent, the measurements were repeated after vasodilatation was induced by hand warming in warm water. Reactive hyperemia was absent in 12 patients when investigations were performed on the unwarmed hand, and these patients had very low resting blood flows. After vasodilatation was induced in these patients, and also in those patients whose resting blood flow was normal, hyperemic responses were comparable in magnitude with those in the controls. The slope of the hyperemic response was significantly less in the patients (1.11 +/- 0.02 V/s) than in the controls (1.28 +/- 0.03 V/s), and therefore, the time course of the hyperemia was lengthened in the patients with SSc, with a delay to achieving maximum blood flow of 2 min. Peak blood flow was directly related to the level of the initial blood flow. These findings support the view that reactive hyperemia is principally a mechanical phenomenon, and also that vessel wall reactivity is abnormal in SSc, producing delayed hyperemic responses. The magnitude of the hyperemia depends on initial flow rates, and the apparent lack of these responses in SSc is a result of their low, but reversible, resting blood flow.


Migraine genetics: a fascinating journey towards improved migraine therapy.
The study of migraine genetics promises to deliver significant changes to migraine therapy. Dr. Ferrari explores the emergence of the field and its importance to headache medicine.


Dysfunction of endothelial progenitor cells under diabetic conditions and its underlying mechanisms.
Cardiovascular complications have been major concerns in the treatment of diabetes, and up to 80% of all deaths in diabetic patients are linked to cardiovascular problems. Impaired angiogenesis is one of the most serious symptoms associated with diabetes, resulting in delayed wound healing and lower limb amputation. Endothelial progenitor cells (EPCs), a subpopulation of adult stem cells, are recruited from bone marrow to the injured vessel to promote endothelial regeneration and neovascularization, playing an important role in angiogenesis. Interestingly, several clinical studies have showed that the number of recruited EPCs is reduced and their function is decreased under diabetic conditions, implying that diabetic EPC dysfunction may contribute to defective angiogenesis and resultant cardiovascular complications in diabetes. To recover the functional abilities of diabetic EPCs and to address possible application of EPC cell therapy to diabetic patients, some studies provided explanations for diabetic EPC dysfunction including increased oxidative stress, involvement of the inflammatory response, alteration in the nitric oxide pathway and reduced signals for EPC recruitment. This review discusses clinical evidence of impairment of EPC functions under diabetic conditions and the suggested mechanisms for diabetic EPC dysfunction.


The use of an overlay autogenous tissue (OAT) patch to control intra-operative haemorrhage of the spleen.
During major abdomino-pelvic surgery, there is a risk of severe haemorrhage. When routine measures at haemostasis fail, the overlay autogenous tissue (OAT) patch may be useful to control bleeding. We present a case report of the use of the use of an OAT patch to control haemorrhage of the spleen incurred during tertiary cytoreduction. The OAT patch should be considered in situations where bleeding is unable to be controlled after routine measures have failed and in instances other than for vascular injury alone. This technique has implications for practice in a wide range of gynaecologic oncology surgery.


[Comparative evaluation of the marginal accuracy of single crowns fabricated computer using aided design/computer aided manufacturing methods, self-curing resin and Luxatemp].
This study aims to compare the marginal accuracy of single crowns fabricated using self-curing resin, Luxatemp, and computer aided design/computer aided manufacturing (CAD/CAM) methods in clinical application. A total of 30 working dies, which were obtained from 30 clinical teeth prepared with full crown as standard, were created and made into 30 self-curing resin, Luxatemp, and CAD/CAM single crowns. The restorations were seated on the working dies, and stereomicroscope was used to observe and measure the thickness of reference points. One-way analysis of variance, which was performed using SPSS 19.0 software package, compared the marginal gap widths of self-curing resin, Luxatemp, and CAD/CAM provisional crowns. The mean marginal gap widths of the fabricated self-curing resin, Luxatemp, and CAD/CAM were (179.06±33.24), (88.83±9.56), and (43.61±7.27) μm, respectively. A significant difference was observed among the three provisional crowns (P<0.05). The marginal gap width of CAD/CAM provisional crown was lower than that of the self-curing resin and Luxatemp. Thus, the CAD/CAM provisional crown offers a better remediation effect in clinical application.


Inhibition of tissue transglutaminase attenuates lipopolysaccharide-induced inflammation in glial cells through AKT/mTOR signal pathway.
In view of the facts that tTG protein expression level and its enzyme activity increase in AD brains of both individuals and transgenic animals and compelling evidence of the involvement of inflammation in AD pathogenesis, tTG could be involved in the inflammation responses in the brain. In the present study, we examined the effects of the irreversible and the competitive inhibitor of tTG on the condition of lipopolysaccharide-induced mimic inflammation models in glial cells. Western blot and tTG enzyme activity assay were applied to detect tTG and isopeptide protein levels and tTG enzyme activity. The production of nitric oxide and the expression levels of inducible nitric oxide synthase and cyclooxygenase-2 were determined by Griess Reagents and Western blot respectively to assess anti-inflammatory effects. Moreover, the activation of AKT/mTOR signaling pathway was determined to evaluate the underlying mechanism of anti-inflammatory response. Irreversible and competitive inhibitor of tTG could ameliorate LPS-induced neuroinflammation in glial cells without cytotoxicity. Moreover, AKT/mTOR pathway may be involved in the anti-inflammatory response of tTG inhibitors. Therefore, NTU283 and Cystamine may alleviate inflammatory response in glial cells, probably through, at least partially, inhibiting the activation of AKT/mTOR signaling pathway. Our study provided some clues that tTG inhibitors NTU283 and Cystamine might be potential candidates for the treatments of neuroinflammation-related diseases, although more studies needed for further exploration.


Na,K-ATPase α2 activity in mammalian skeletal muscle T-tubules is acutely stimulated by extracellular K+.
The Na,K-ATPase α2 isoform is the predominant Na,K-ATPase in adult skeletal muscle and the sole Na,K-ATPase in the transverse tubules (T-tubules). In quiescent muscles, the α2 isozyme operates substantially below its maximal transport capacity. Unlike the α1 isoform, the α2 isoform is not required for maintaining resting ion gradients or the resting membrane potential, canonical roles of the Na,K-ATPase in most other cells. However, α2 activity is stimulated immediately upon the start of contraction and, in working muscles, its contribution is crucial to maintaining excitation and resisting fatigue. Here, we show that α2 activity is determined in part by the K+ concentration in the T-tubules, through its K+ substrate affinity. Apparent K+ affinity was determined from measurements of the K1/2 for K+ activation of pump current in intact, voltage-clamped mouse flexor digitorum brevis muscle fibers. Pump current generated by the α2 Na,K-ATPase, Ip, was identified as the outward current activated by K+ and inhibited by micromolar ouabain. Ip was outward at all potentials studied (-90 to -30 mV) and increased with depolarization in the subthreshold range, -90 to -50 mV. The Q10 was 2.1 over the range of 22-37°C. The K1/2,K of Ip was 4.3±0.3 mM at -90 mV and was relatively voltage independent. This K+ affinity is lower than that reported for other cell types but closely matches the dynamic range of extracellular K+ concentrations in the T-tubules. During muscle contraction, T-tubule luminal K+ increases in proportion to the frequency and duration of action potential firing. This K1/2,K predicts a low fractional occupancy of K+ substrate sites at the resting extracellular K+ concentration, with occupancy increasing in proportion to the frequency of membrane excitation. The stimulation of preexisting pumps by greater K+ site occupancy thus provides a rapid mechanism for increasing α2 activity in working muscles.


Patterns of cervical and masticatory impairment in subgroups of people with temporomandibular disorders-an explorative approach based on factor analysis.
To identify clinical patterns of impairment affecting the cervical spine and masticatory systems in different subcategories of temporomandibular disorder (TMD) by an explorative data-driven approach. For this observational study, 144 subjects were subdivided according to Research Diagnostic Criteria for TMDs into: Healthy controls, temporomandibular joint (TMJ) signs without symptoms, TMJ affected, temporomandibular muscles affected, or TMJ and muscles affected. Factor analysis and linear regression were applied to cervical spine and masticatory data to identify and characterize clinical patterns in subgroups. Factor analysis identified five clinical dimensions, which explained 59% of all variance: Mechanosensitivity, cervical movement, cervical and masticatory dysfunction, jaw movement, and upper cervical movement. Regression analysis identified different clinical dimensions in each TMD subgroup. Distinct clinical patterns of cervical spine and masticatory function were found among subgroups of TMD, which has clinical implications for therapeutic management.


Synthetically modified mRNA for efficient and fast human iPS cell generation and direct transdifferentiation to myoblasts.
Synthetic mRNA transfection enables efficient and controlled gene expression in human cells, without genome integrations. Further, modifications to the mRNA and transfection protocol now allow for repeated transfection and long-term gene expression of an otherwise short-lived mRNA expression. This is mainly achieved through introducing modified nucleosides and interferon suppression. In this study we provide an overview and details of the advanced synthesis and modifications of mRNA originally developed for reprogramming. This mRNA allows for very efficient transfection of fibroblasts enabling the generation of high quality human iPS cells with a six-factor mRNA cocktail in 9 days. Furthermore, we synthesised and transfected modified MYOD1 mRNA to transdifferentiate human fibroblasts into myoblast-like cells without a transgene footprint. This efficient and integration-free mRNA technology opens the door for safe and controlled gene expression to reverse or redirect cell fate.


Programmed chest-wall stimulation to evaluate the progress of A-V block after pacemaker insertion in patients with trifascicular disease.
Twenty six patients (aged 46-80, mean age 64) with bifascicular block in the presence of prolonged H-V interval (trifascicular block), were followed for an average of 31 months after inserting an R-wave inhibited pacemaker (PM) because of syncope and/or dizzy attacks. The underlying rhythm was evaluated at 4-6-month intervals by three different techniques: 1) 12-lead ECG when intrinsic patient rate was faster than PM rate; 2) abrupt PM inhibition (APMI) by the rapid chest-wall stimulation technique, and 3) progressive PM inhibition (PPMI) using a programmed chest-wall stimulation technique capable of decreasing the PM rate gradually to 30 beats/min before complete PM inhibition. In addition, the PPMI allowed the underlying rhythm to be induced and sustained and properly evaluated without any discomfort to the patient. Following PM insertion, 4 patients (15%) developed complete heart block after a mean follow-up of 43 months, and one patient (4%) developed 2nd degree 2:1 A-V block (VX) after 83 months. The P-R interval increased in 5 patients (19%) and decreased in 2 (8%). No change of A-V conduction was found in 9 patients (34%). Three patients developed low atrial rhythm, atrial flutter and atrial fibrillation, respectively (12%). After PM insertion 2 patients still complained of dizziness. None reported syncope. Two patients died during follow-up, both of congestive heart failure (8%). By detection of intrinsic rhythm it was recognized that a long symptomatic paroxysmal phase may precede the development of chronic complete A-V block. Therefore, the insertion of a permanent PM is recommended in patients with unexplained neurologic symptoms and trifascicular disease, without waiting for documented episodes of complete A-V block.


Intracellular distribution of molecular forms of acetylcholinesterase in rat brain and changes after diisopropylfluorophosphate treatment.
The subcellular distribution of acetylcholinesterase activities was studied in the striatum and cerebellum of rat brain. The highest percentage of the enzyme activity was found in the crude synaptosomal (P2) fraction, with striatum much higher than cerebellum. On sucrose density gradient centrifugation analyses all the particulate fractions (P1, P2, and P3) showed a major peak of the 10 S form of acetylcholinesterase activity with very little activity of the 4 S form of the enzyme. The 10 S/4 ratio was much higher in striatum than in cerebellum. In the soluble fraction (100,000 g supernatant) the 10 S form was less than the 4 S form in the adult rat brain, but this was reversed in the 6-day-old rat brain. After diisopropylfluorophosphate administration the recovery of acetylcholinesterase molecular forms in various subcellular fractions differed at different recovery periods. These results indicate that the distribution of molecular forms of acetylcholinesterase in rat brain differs in various subcellular fractions, and also the pattern of distribution differs in different regions of the brain as well as in adult and developing brains.


Otitis media, tympanostomy tube placement, and use of antibiotics. Cross-sectional community study repeated after five years.
To investigate potential links between antimicrobial drug use for acute otitis media (AOM) and tympanostomy tube placements, and the relationship between parental views and physician antimicrobial prescribing habits. Cross-sectional community study repeated after five years. Representative samples of children aged 1-6 years in four well-defined communities in Iceland, examined in 2003 (n = 889) and 1998 (n = 804). Prevalence of antimicrobial treatments for AOM, tympanostomy tube placements, and parental expectations of antimicrobial treatment. Results. Tympanostomy tubes had been placed at some time in 34% of children in 2003, as compared with 30% in 1998. A statistically significant association was found between tympanostomy tube placement rate and antimicrobial use for AOM in 2003. In the area where antimicrobial use for AOM was lowest in 1998, drug use had further diminished significantly. At the same time, the prevalence of tympanostomy tube placements had diminished from 26% to 17%. Tube placements had increased significantly, from 35% to 44%, in the area where antimicrobial use for AOM was highest. Parents in the area where antimicrobial consumption was lowest and narrow spectrum antimicrobials were most often used were less likely to be in favour of antimicrobial treatment. Comparison between communities showed a positive correlation between antimicrobial use for AOM and tympanostomy tube placements. The study supports a restrictive policy in relation to prescriptions of antibiotics for AOM. It also indicates that well-informed parents predict a restrictive prescription policy.


Development of chicken epidermis cultured with embryo extract.
The epidermis from 11-day-old chick embryo shank skin was cultured with 11-day-old chick embryo extract. The growth and the differentiation of the epidermis in culture were studied histologically, electron microscopically and with polyacrylamide gel electrophoresis of keratin proteins. The epidermis cultured with the chick embryo extract proliferated and stratum structures developed simultaneously with the increase in epidermal cell layers. Finally, a keratinized layer was observed after 10 days in culture. Electron microscopic observations revealed that tonofilaments were produced after 2 days in culture and increased thereafter with culture time, becoming condensed with desmosomes. Keratohyaline granules were observed in 7-day cultures. These keratinization characteristics occurring during culture showed the general characteristics of the alpha stratum observed in the skin of in ovo embryos during the early stages of development. However, the development of peridermal and subperidermal granules was poor and the stratum granulosum, which develops at the late stages between the stratum intermedium and the stratum corneum, was not observed. Polyacrylamide gel electrophoresis of S-carboxymethylated keratin proteins showed that the keratin protein band patterns of the culture differed from those of in ovo skin epidermis.


Insight into broad substrate specificity and synergistic contribution of a fungal α-glucosidase in Chinese Nong-flavor daqu.
Chinese Nong-favor daqu, the presentative liquor starter of Baijiu, has been enriched with huge amounts of enzymes in degrading various biological macromolecules by openly man-made process for thousand years. According to previous metatranscriptomics analysis, plenty of α-glucosidases were identified to be active in NF daqu and played the key role in degrading starch under solid-state fermentation. However, none of α-glucosidases was characterized from NF daqu, and their actual functions in NF daqu were still unknown. An α-glucosidase (NFAg31A, GH31-1 subfamily), the second highest expressed α-glucosidases in starch degradation of NF daqu, was directly obtained by heterologous expression in Escherichia coli BL21 (DE3). NFAg31A exhibited the highest sequence identities of 65.8% with α-glucosidase II from Chaetomium thermophilum, indicating its origin of fungal species, and it showed some similar features with homologous α-glucosidase IIs, i.e., optimal activity at pH ~ 7.0 and litter higher temperature of 45 ℃, well stability at 41.3 ℃ and a broad pH range of pH 6.0 to pH 10.0, and preference on hydrolyzing Glc-α1,3-Glc. Besides this preference, NFAg31A showed comparable activities on Glc-α1,2-Glc and Glc-α1,4-Glc, and low activity on Glc-α1,6-Glc, indicating its broad specificities on α-glycosidic substrates. Additionally, its activity was not stimulated by any of those detected metal ions and chemicals, and could be largely inhibited by glucose under solid-state fermentation. Most importantly, it exhibited competent and synergistic effects with two characterized α-amylases of NF daqu on hydrolyzing starch, i.e., all of them could efficiently degrade starch and malto-saccharides, two α-amylases showed advantage in degrading starch and long-chain malto-saccharides, and NFAg31A played the competent role with α-amylases in degrading short-chain malto-saccharides and the irreplaceable contribution in hydrolyzing maltose into glucose, thus alleviating the product inhibitions of α-amylases. This study provides not only a suitable α-glucosidase in strengthening the quality of daqu, but also an efficient way to reveal roles of the complicated enzyme system in traditional solid-state fermentation. This study would further stimulate more enzyme mining from NF daqu, and promote their actual applications in solid-state fermentation of NF liquor brewing, as well as in other solid-state fermentation of starchy industry in the future.


Transcriptional Fluctuations Govern the Serum-Dependent Cell Cycle Duration Heterogeneities in Mammalian Cells.
Mammalian cells exhibit a high degree of intercellular variability in cell cycle period and phase durations. However, the factors orchestrating the cell cycle duration heterogeneities remain unclear. Herein, by combining cell cycle network-based mathematical models with live single-cell imaging studies under varied serum conditions, we demonstrate that fluctuating transcription rates of cell cycle regulatory genes across cell lineages and during cell cycle progression in mammalian cells majorly govern the robust correlation patterns of cell cycle period and phase durations among sister, cousin, and mother-daughter lineage pairs. However, for the overall cellular population, alteration in the serum level modulates the fluctuation and correlation patterns of cell cycle period and phase durations in a correlated manner. These heterogeneities at the population level can be fine-tuned under limited serum conditions by perturbing the cell cycle network using a p38-signaling inhibitor without affecting the robust lineage-level correlations. Overall, our approach identifies transcriptional fluctuations as the key controlling factor for the cell cycle duration heterogeneities and predicts ways to reduce cell-to-cell variabilities by perturbing the cell cycle network regulations.


Isolation of endophytic fungi and screening of Huperzine A-producing fungus from Huperzia serrata in Vietnam.
Huperzine A (HupA), a natural Lycopodium alkaloid derived from Huperzia serrata (Thunb. ex Murray) Trev. plants, is a highly active acetylcholinesterase inhibitor and a key compound used for treating Alzheimer's disease (AD). Recently, HupA has been reported in various endophytic fungi isolated from H. serrata. In the present study, 153 endophytic fungi were isolated from healthy tissues of H. serrata collected from natural populations in Lam Dong province of Central Vietnam. The endophytic fungi were identified based on morphological characteristics and Internal Transcribed Spacer sequences. Among them, 34 strains were classified into seven genera belonging to Ascomycota, including Alternaria, Fusarium, Trichoderma, Penicillium, Paecilomyces, and Phoma, and eight strains belonging to the genus Mucor (Zygomycota). The other strains remained unidentified. According to the results of thin-layer chromatography and high-performance liquid chromatography, only one of the 153 strains, Penicillium sp. LDL4.4, could produce HupA, with a yield 1.38 mg l-1 (168.9 µg g-1 dried mycelium) when cultured in potato dextrose broth, which was considerably higher than that of other reported endophytic fungi. Such a fungus is a promising candidate and alternative to presently available HupA production techniques for treating AD and preventing further memory decline.


[Liposomal curcumin inhibits tumor growth and angiogenesis in Lewis lung cancer].
To prepare water soluble curcumin liposome and investigate its anti-tumour and antiangiogenic effects. Liposomal curcumin was prepared by alcohol injection method. Proliferation inhibition to murin Lewis lung cancer cell line LL/2 of curcumin liposome was evaluated by MTT assay. Apoptosis and cell cycle arrest induced by liposomal curcumin were analysed by flow cytometry. Anti-tumour effects were investigated in a murine lung cancer model, and the anti-angiogenic effect was determined by aginate encapsulation assay. In vitro, liposomal curcumin inhibits the proliferation of LL/2 cells and induces apoptosis and cell cycle arrest. In vivo, the systemic administration of liposomal curcumin resulted in the inhibition of tumour. Aginate encapsulation assay revealed angiogenesis was decreased by curcumin liposome. The curcumin liposome treatment can significantly inhibit tumour growth in vivo.


BMI and Allostatic Load Are Directly Associated with Longitudinal Increase in Plasma Neurofilament Light among Urban Middle-Aged Adults.
Plasma neurofilament light chain (NfL) is a novel biomarker for age-related neurodegenerative disease. We tested whether NfL may be linked to cardiometabolic risk factors, including BMI, the allostatic load (AL) total score (ALtotal), and related AL continuous components (ALcomp). We also tested whether these relations may differ by sex or by race. We used data from the HANDLS (Healthy Aging in Neighborhoods of Diversity across the Life Span) study [n = 608, age at visit 1 (v1: 2004-2009): 30-66 y, 42% male, 58% African American] to investigate associations of initial cardiometabolic risk factors and time-dependent plasma NfL concentrations over 3 visits (2004-2017; mean ± SD follow-up time: 7.72 ± 1.28 y), with outcomes being NfLv1 and annualized change in NfL (δNfL). We used mixed-effects linear regression and structural equations modeling (SM). BMI was associated with lower initial (γ01 = -0.014 ± 0.002, P < 0.001) but faster increase in plasma NfL over time (γ11 = +0.0012 ± 0.0003, P < 0.001), a pattern replicated for ALtotal. High-sensitivity C-reactive protein (hsCRP), serum total cholesterol, and resting heart rate at v1 were linked with faster plasma NfL increase over time, overall, while being uncorrelated with NfLv1 (e.g., hsCRP × Time, full model: γ11 = +0.004 ± 0.002, P = 0.015). In SM analyses, BMI's association with δNfL was significantly mediated through ALtotal among women [total effect (TE) = +0.0014 ± 0.00038, P < 0.001; indirect effect = +0.00042 ± 0.00019, P = 0.025; mediation proportion = 30%], with only a direct effect (DE) detected among African American adults (TE = +0.0011 ± 0.0004, P = 0.015; DE = +0.0010 ± 0.00048, P = 0.034). The positive associations between ALtotal/BMI and δNfL were mediated through increased glycated hemoglobin (HbA1c) concentrations, overall. Cardiometabolic risk factors, particularly elevated HbA1c, should be screened and targeted for neurodegenerative disease, pending comparable longitudinal studies. Other studies examining the clinical utility of plasma NfL as a neurodegeneration marker should account for confounding effects of BMI and AL.


Polymorphisms in DLGH1 and LAMC1 in Mayer-Rokitansky-Kuster-Hauser syndrome.
Müllerian agenesis, also termed the Mayer-Rokitansky-Kuster-Hauser syndrome (MRKHS) is a disorder with an incidence of approximately 1 in 4500 newborn girls. This study screened 12 patients with MRKHS for mutations in two genes, LAMC1 and DLGH1, involved in the development of Müllerian structures and found 10 previously described variants and no novel variants in the coding sequence. It is highly unlikely that these variants are pathological since these are common in the general population. It is the first time that an extensive study of LAMC1 and DLGH1 has been undertaken in patients with MRKHS. The data support the notion that mutations in the coding sequence of LAMC1 and DLGH1 may not be associated with MRKHS.


Effect of vaginal microbiota on pregnancy outcomes of women from Northern China who conceived after IVF.
This study aimed to investigate the correlation between vaginal microbiota and pregnancy outcomes of women who achieved pregnancy via in vitro fertilization (IVF) in Northern China, and to determine a biomarker for evaluation of the risk of preterm births in these women. In total, 19 women from Northern China women who conceived after IVF and 6 women who conceived naturally were recruited in this study. The vaginal samples of the healthy participants were collected throughout pregnancy, that is, during the first, second, and third trimesters. The V3-V4 region of 16S rRNA was used to analyze the vaginal microbiome, and the bioinformatic analysis was performed using QIIME Alpha and Beta diversity analysis. Either IVF group or Natural conception group, bacterial community diversities and total species number of vagnal samples from who delivered at term were significantly higher than those who delivered before term. Low abundance of vaginal bacteria indicates an increased risk of preterm delivery. Further, more abundant vaginal bacteria was found in first trimesters instead of the next two trimesters. Vignal samples collected during first trimester showed richer differences and more predictive value for pregnancy outcoes. In addition, the diversity of the vaginal bacterial community decreased as the gestational age increased, in all samples. Alloscardovia was only found in participants who conceived after IVF, and the percentage of Alloscardovia in viginal samples of normal delivery group is much higher than the samples from preterm delivery group.Vobrio specifically colonized in vagina of pregnant woman in AFT group (those who conceived after IVF (A), first trimester (F), and delivered at term (T)) and Sporosarcina was detected only in women with AFT and AST (those who conceived after IVF (A), second trimester (S), and delivered at term (T)). These data indicates that Alloscardovia, Vobrio and Sporosarcina have great potential in predicting pregnancy outcomes who pregnanted by vitro fertilization. Vaginal microbiota were more stable in women who conceived naturally and those who carried pregnancy to term. Oceanobacillus might act as a positive biomarker, whereas Sulfurospirillum and Propionispira may act as negative biomarkers for the risk of preterm birth.


Evaluation of the predictivity of Acute Oral Toxicity (AOT) structure-activity relationship models.
Several public efforts are aimed at discovering patterns or classifiers in the high-dimensional bioactivity space that predict tissue, organ or whole animal toxicological endpoints. The current study sought to assess and compare the predictions of the Globally Harmonized System (GHS) categories and Dangerous Goods (DG) classifications based on Lethal Dose (LD50) from several available tools (ACD/Labs, Leadscope, T.E.S.T., CATMoS, CaseUltra). External validation was done using dataset of 375 substances to demonstrate their predictive capacity. All models showed very good performance for identifying non-toxic compounds, which would be useful for DG classification, developing or triaging new chemicals, prioritizing existing chemicals for more detailed and rigorous toxicity assessments, and assessing non-active pharmaceutical intermediates. This would ultimately reduce animal use and improve risk assessments. Category-to-category prediction was not optimal, mainly due to the tendency to overpredict the outcome and the general limitations of acute oral toxicity (AOT) in vivo studies. Overprediction does not specifically pose a risk to human health, it can impact transport and material packaging requirements. Performance for compounds with LD50 ≤ 300 mg/kg (approx. 5% of the dataset) was the poorest among all groups and could be potentially improved by including expert review and read-across to similar substances.


Body temperature change during anesthesia for electroconvulsive therapy: implications for quality incentives in anesthesiology.
The American Society of Anesthesiologists has announced that perioperative normothermia is a "Quality Incentive in Anesthesiology." We examined whether we could meet this quality incentive in a simple population: patients undergoing anesthesia for electroconvulsive therapy (ECT). We compared infrared-measured ear temperature before anesthesia to temperature upon delivery of patients to the postanesthesia care unit (PACU) after 101 consecutive brief anesthetics to facilitate ECT. For 35 procedures, the patients had an infrared ear thermometer temperature of <36 degrees C before anesthesia was administered, and 18 had a temperature of <36 degrees C after anesthesia when transferred to the PACU. For 30 anesthetics, the patients' temperature decreased during anesthesia, for 64 anesthetics it increased during anesthesia, and for 7 it did not change. Overall examination of the data demonstrated no correlation between preprocedure and postprocedure temperature. We conclude that there was no consistent change in temperature during anesthesia between our study patients when anesthesia was administered to facilitate ECT. If patients' tympanic temperatures were below 36 degrees C upon admission to the PACU, it would be incorrect to conclude that intraprocedural temperature management measures were substandard. Also, current methods of measuring temperature may be inadequate to ascertain if patients are hypothermic after surgery. As the avoidance of hypothermia is a meritorious goal, anesthesia departments need to ensure that their temperature monitoring equipment is adequate to ensure accurate measurement of postanesthetic temperature if this variable is to be used as a quality incentive.


Future Offspring Costs in Economic Evaluation.
Economic evaluation guidelines increasingly prescribe inclusion of all future costs. We point at an important dimension of future costs that is systematically neglected. Healthcare can affect future offspring, either through affecting the patient's fertility or through determining future offspring's health. As we show, the future costs associated with these changes can be substantial and will vary across interventions and demographic groups. However, systematic inclusion of these future offspring costs would raise many problems on its own. Based on the population ethics concept of necessitarianism, we suggest that only those future costs that spring from 'necessary' future lives should be included in future cost calculations, while all costs associated with 'potential' future lives can be ignored. This approach allows excluding most future offspring costs and avoids skewed cost-effectiveness outcomes of interventions with fertility effects, while taking into account the economic implications of preventing disease in future generations that will exist by necessity. Overall, future generations expose a substantial gap in today's Health Technology Assessment (HTA) methodology and further discussion of the issues they raise is needed.


Impact of the COVID-19 Pandemic on Early Career Investigators in Rheumatology: Recommendations to Address Challenges to Early Research Careers.
The COVID-19 pandemic has impacted the careers of trainees and early career investigators (ECIs). We sought to assess how the American College of Rheumatology (ACR) and the Rheumatology Research Foundation (RRF) can address the needs of those pursuing research careers. The Committee on Research created a survey to assess the impact of COVID-19 and identify topics for the ACR and the RRF to address. In fall of 2020, we surveyed postdoctoral trainees and ECIs within 9 years of terminal training. Responses were analyzed using descriptive statistics and qualitative content analysis. Twenty-one percent of invitees responded to the survey (n = 365); of these, 60% were pursuing careers in academic research. Seventy-five percent of respondents in academic research career paths placed their primary projects on hold during the pandemic. The number of individuals pursuing a research career from 2020 to 2021 decreased by 5%. Respondents reported funding, caregiving, and lack of preliminary data as significant challenges. Suggested impactful interventions included increased funding, funding process reform, and expanding mentoring and networking resources. Major stressors identified during the pandemic included increased caregiving responsibilities and difficulty obtaining data and funding, for which respondents suggested increases and changes in funding programs as well as more mentoring and networking opportunities. Based on these, the Committee on Research proposes 3 priorities: 1) flexible funding mechanisms for ECIs and additional support for those impacted by caregiving; 2) virtual and in-person programs for career development and networking; and 3) curated content relevant to building a research career available on demand.


Prevention of Radiation-Induced Bladder Injury: A Murine Study Using Captopril.
Pelvic radiation therapy (RT) can cause debilitating bladder toxicities but few clinical interventions exist to prevent injury or alleviate symptoms. From a large genome-wide association study in patients with prostate cancer it was previously reported that SNPs tagging AGT, part of the renin-angiotensin system (RAS), correlated with patient-reported late hematuria, identifying a potential targetable pathway to prevent RT-induced bladder injury. To investigate this association, we performed a preclinical study to determine whether RAS modulation protected the bladder against RT injury. C57BL/6 male mice were treated with an oral angiotensin converting enzyme inhibitor (ACEi: 0.3g/L captopril) 5 days before focal bladder X-irradiation with either single dose (SD) 30 Gy or 3 fractions of 8 Gy (8 Gy × 3 in 5 days). RT was delivered using XStrahl SARRP Muriplan CT-image guidance with parallel-opposed lateral beams. ACEi was maintained for 20 weeks post RT. Bladder toxicity was assessed using assays to identify local injury that included urinalysis, functional micturition, bladder-released exosomes, and histopathology, as well as an assessment of systemic changes in inflammatory-mediated circulating immune cells. SD and fractionated RT increased urinary frequency and reduced the volume of individual voids at >14 weeks, but not at 4 weeks, compared with nonirradiated animals. Urothelial layer width was positively correlated with mean volume of individual voids (P = .0428) and negatively correlated with number of voids (P = .028), relating urothelial thinning to changes in RT-mediated bladder dysfunction. These chronic RT-induced changes in micturition patterns were prevented by captopril treatment. Focal bladder irradiation significantly increased the mean particle count of urine extracellular vesicles and the monocyte and neutrophil chemokines CCL2 and MIP-2, and the proportions of circulating inflammatory-mediated neutrophils and monocytes, which was also prevented by captopril. Exploratory transcriptomic analysis of bladder tissue implicated inflammatory and erythropoietic pathways. This study demonstrated that systemic modulation of the RAS protected against and alleviated RT-induced late bladder injury but larger confirmatory studies are needed.


[RELATIONSHIP BETWEEN BONE MINERAL DENSITY AND LUMBAR INTERVERTEBRAL DISC DEGENERATION IN RHESUS MACAQUES].
To analyze the relationship between the bone mineral density (BMD) and lumbar intervertebral disc degeneration in rhesus macaques by using T1ρ-MRI. Twenty female rhesus macaques at the age of 10.9 years on average (rang, 4-20 years) were selected. The lumbar intervertebral discs were classified by Pfirrmann grading system and the T1ρ relaxation time (T1ρ value) was examined by using MRI (Philips 1.5 Tesla), and then BMD values of the L4,5 vertebrae and femoral ward's triangle were detected by using Osteocore dual energy X-ray absorptiometry. Finally, the relationship of T1ρ value of the lumbar intervertebral discs and Pfirrmann grading with age, weight, BMD of lumbar vertebrae and femoral ward's triangle was analyzed. The BMD values of lumbar vertebrae and femoral ward's triangle were (0.64 ± 0.17) g/cm2 and (0.67 ± 0.19) g/cm2 respectively, showing no significant difference (t = 2.893, P = 0.128). According to Pfirrmann grading system, there were 7 cases of grade I, 8 cases of grade II, and 5 cases of grade III at L4,5 intervertebral discs. The T1ρ value of the lumbar intervertebral disc was (104.08 ± 18.65) ms; the T1ρ values of grades I, II, and III were (121.31 ± 13.44), (104.73 ± 15.01), and (77.41 ± 11.87) ms, respectively. There was a negative correlation between T1ρ value and the age and the BMD of lumbar vertebrae and femoral ward's triangle. There was a positive correlation between Pfirrmann grading and the variables as listed above. Significant negative linear correlation was also observed between T1ρ value and Pfirrmann grading. The T1ρ value is a reliable index when quantifying lumbar intervertebral disc degeneration, and there is a significant positive correlation between BMD and lumbar intervertebral disc degeneration in rhesus macaques.


Alcohol use in television programming: effects on children's behavior.
The impact of televised use of alcoholic beverages on children's behavior was evaluated in this study. Children were randomly assigned to one of three experimental conditions: (a) television with drinking depicted, (b) television without drinking, or (c) a no television control condition. Segments of the syndicated television show M.A.S.H. were used as the viewing stimulus. The impact of the various conditions was evaluated in a choice task requiring subjects to choose either martini or regular glasses of either "whiskey" or "water" to serve pictured adults and children. Results indicated that subjects who viewed television drinking were significantly more likely to choose alcoholic beverages for pictured adults but not children. No effect was found for glass preference. Results and implications of these findings and directions for future research are discussed.


Cerebellar infarction secondary to subclavian aortoplasty repair for coarctation of the aorta.
A five-year-old boy developed a left cerebellar infarction following repair of coarctation of the aorta by subclavian aortoplasty. At operation a large left vertebral artery had been ligated. If a large vertebral artery is encountered at repair of coarctation of the aorta then consideration should be given to a method of repair which does not sacrifice this vessel.