Ca2+ dependence of alpha-adrenergic effects on the contractile properties and Ca2+ homeostasis of cardiac myocytes.
alpha-Adrenergic stimulation is known to enhance myocardial contractility. Adult rat left ventricular myocytes bathed in 1 mM [Ca2+] (Ca0) and electrically stimulated at 0.2 Hz responded to alpha-adrenergic stimulation with 50 microM phenylephrine and 1 microM propranolol with an increase in twitch amplitude to 177.1 +/- 25.6% of control (mean +/- SEM). In contrast, when cell Ca2+ loading was increased by bathing cells in 5 mM Ca0, alpha-adrenergic stimulation decreased twitch amplitude to 68.6 +/- 8.2% of control. Time-averaged cytosolic [Ca2+] of cells in 1.0 mM Ca0 is enhanced via an increase in the frequency of electrical stimulation. When myocytes were stimulated at 2 Hz in 1 mM Ca0, alpha-adrenergic stimulation did not increase twitch amplitude (103.8 +/- 12.4% of control). In myocytes loaded with the Ca2+ probe into-1, alpha-adrenergic effects during stimulation at 0.2 Hz (an increase in twitch amplitude in 1 mM Ca0 and a decrease in twitch amplitude in 5 mM Ca0) were associated with similar changes in the indo-1 transient. In 5 mM Ca0, spontaneous Ca2+ releases from the sarcoplasmic reticulum (SR) occurred in the diastolic interval between twitches (2.9 +/- 1.4 spontaneous SR Ca2+ oscillations/min; n = 7); alpha-adrenergic stimulation abolished these oscillations in six of seven cells. Thus, an increase in the frequency of spontaneous diastolic SR Ca2+ release (i.e., Ca2+ overload) is not the mechanism for the negative inotropic effect of alpha-adrenergic stimulation in 5 mM Ca0. In experiments with unstimulated myocytes, we determined whether the effect of alpha-adrenergic stimulation on cell Ca2+ homeostasis and oscillatory SR Ca2+ release observed in 5 mM Ca0 occurs only during electrical stimulation, when voltage-dependent currents are operative, or also at rest. Unstimulated rat ventricular myocytes in 5 mM Cao exhibit oscillatory SR Ca2+ release; alpha-adrenergic stimulation decreased the frequency of these oscillations to 53.9 +/- 8.9% of control, and this effect was blocked by 1 microM prazosin. In unstimulated indo-1-loaded myocytes alpha-adrenergic stimulation decreased the resting indo-1 fluorescence ratio in 5 mM Ca0, whereas it had no effect in 1 mM Ca0. Additional experiments were aimed at defining a role for Ca(2+)-activated, phospholipid-dependent protein kinase C (PKC) for the negative inotropic effect of alpha-adrenergic stimulation in 5 mM Ca0. Short-term preexposure to 0.1 microM 4 beta-phrobol 12-myristate 13-acetate (PMA) has been shown to maximally activate PKC.(ABSTRACT TRUNCATED AT 400 WORDS)


Complement fixing and lymphocytotoxic antibodies in serum of pregnant women at delivery. III. Clinical observations.
The investigation comprised 2,008 pregnancies. Sera fnd lymphocytotoxic antibodies (LCTAb). 14 women had only CFPAb, 44 had CFPAb and LCTAb, 224 had only LCTAb. In 3 of 481 cord blood serum samples LCTAb were demonstrated. No cases of neutropenia or thrombocytopenia was observed. No association was found between CFPAb or LCTAb and stillbirth, birth defects, degree of bilirubinaemia, diseases of the newborn, or placental weight. It was suggested that the applied lymphocytotoxicity test detects both in vivo cytolytic antibodies and enhancing antibodies, which may explain the results.


Disability, impairment, and physical therapy utilization after arthroscopic partial meniscectomy in patients receiving workers' compensation.
The treatment of meniscal tears in injured workers is associated with less favorable outcomes and higher utilization of clinical services. It was hypothesized that patients receiving Workers' Compensation who undergo arthroscopic meniscectomy can have excellent outcomes with physical therapy utilization below the national best-practices benchmarks. The records of 155 injured workers who had undergone 164 primary arthroscopic meniscectomies were reviewed at least one year following claim closure. The time to release to unrestricted full work duty and the number of postoperative physical therapy visits were compared between the study group (managed with a protocol-driven, independent exercise program) and the control group (managed with traditional outpatient physical therapy). The traditional therapy regimen was implemented by means of a written referral stating general goals (knee range of motion, strength, and function) and recommending a range or a maximum number of visits to be attended. Patients in the study group received a written referral specifying the exact number of visits that were approved, a kit containing exercise equipment with a booklet illustrating twenty-five exercises, and a prescribed rehabilitation protocol outlining the philosophy, expected subjective and functional outcomes, and specific objective weekly goals. The median number of physical therapy visits per patient was 40% lower in the study group than in the control group (six compared with ten; p < 0.001). There was no difference between the study group and the control group with regard to the permanent partial disability rate (0% compared with 4.3%; p = 0.076). Following arthroscopic meniscectomy, there was no significant difference between the study group and the control group in terms of the time to release to light duty, the time to release to full duty, the time to claim closure, or the rate of impairment. The implementation of a structured, independent exercise protocol appears promising as a method to reduce physical therapy utilization to levels below the national best-practices benchmarks without negatively impacting impairment and disability rates for patients receiving Workers' Compensation who undergo arthroscopic meniscectomy.


[Space-time sequence and mosaicism of neurogenesis in the CA1 field of the mouse hippocampus].
Pregnant CBA mice were given a single injection of 3H-thymidine (3H-T) (10 microCi/g) on days 11-19 of pregnancy. The mouse progeny was sacrificed on the first day of postnatal life. Brain was embedded into paraffin and durcupan. Radioautographs of paraffin and semithin sections were prepared and employed for mapping the site of intensely labeled neurons (ILN) in the CAI area of the hippocampus (H). ILN appeared in the CAI area after 3H-T injection, namely on embryonic day 12 (E 12). The ILN number reached a maximum after isotope injection on 2 14-15 and then dramatically fell down. The neurogenesis in the suprapyramidal layer of the CAI area slightly outstripped the neurogenesis in the pyramidal and infrapyramidal layers. At early times of the experiment the CAI area exhibited the predominance of single ILN, whereas at late times paired ILN prevailed. That fact might be linked with the replacement during the neurogenesis of asymmetric critical mitoses of the germinative neuronal precursors by symmetric critical mitoses. Analysis of the ILN distribution in the CAL area revealed mosaic clusters of ILN alternating with unlabeled or mildly labeled neurons. Those groups were most remarkable in mice injected with 3H-T on E 14 and E 15. The mosaicism of the neurogenesis in the H is regarded as the result of heterochronous neuronal production by local parts of the germinative zone, each of which builds up a separate radial segment of the H.


Resonance offset tailored composite pulses.
We describe novel composite pulse sequences which act as general rotors and thus are particularly suitable for nuclear magnetic resonance quantum computation. The resonance offset tailoring to enhance nutations approach permits perfect compensation of off-resonance errors at two selected frequencies placed symmetrically around the frequency of the radiofrequency source.


De novo developed microsatellite markers in gill parasites of the genus Dactylogyrus (Monogenea): Revealing the phylogeographic pattern of population structure in the generalist parasite Dactylogyrus vistulae.
Approaches using microsatellite markers are considered the gold standard for modern population genetic studies. However, although they have found application in research into various platyhelminth taxa, they remained substantially underutilized in the study of monogeneans. In the present study, a newly developed set of 24 microsatellite markers was used to investigate the genetic diversity of the generalist monogenean species Dactylogyrus vistulae. The analyzed parasite specimens were collected from 13 cyprinoid species from 11 sites in the Apennine and Balkan peninsulas. A total of 159 specimens were genotyped at each of the loci and the number of alleles per locus ranged from 2 to 16, with a mean number of 6.958 alleles per locus. Exceptionally high genetic diversity was observed among D. vistulae individuals in the southern Balkans (mean N A per locus = 3.917), suggesting that generalist D. vistulae expanded from the south to the north in the Balkans and later into central Europe. The initial clustering analysis divided all investigated specimens into three major clusters; however, the results of the subsequent analyses revealed the existence of various subpopulations, suggesting that the population structure of D. vistulae is associated with the diversification of their cyprinoid hosts. In addition, the partition of the parasite population was observed in regions of the sympatric occurrence of two host species, indicating that these hosts may represent a barrier for gene flow, even for generalist parasite species.


Compact multi-functional frequency-selective absorber based on customizable impedance films.
Multi-functional metamaterial absorbers have attracted considerable attention for applications in the microwave frequency regime. In this paper, we report the design, fabrication, and characterization of frequency-selective absorbers, which exhibit substantial absorption property within a pre-defined frequency band, while at the same time behaving as a highly transparent screen in another targeted frequency band. The proposed designs consist of a symmetrically patterned indium tin oxide film acting as an absorbing layer, two dielectric substrates, and a cross-slot metal sheet frequency selective surface playing the role of a transmitting layer. In order to validate the functionalities of the designed absorbers, equivalent circuit models, full-wave numerical simulations and measurements are presented. The measured results, in good agreement with the numerical ones, show that the proposed designs realize 80% broadband absorption over the desired frequency range and possess a transparent window in a higher or lower frequency band for a wide range of incidence angles up to 60°. These performances suggest that the proposed designs are promising candidates for multi-functional scattering control and communication applications.


Muscle fibre direction of longissimus, iliocostalis and multifidus: landmark-derived reference lines.
Considerable inter-individual variations in the fibre direction angles of the iliocostalis lumborum, longissimus and multifidus were observed, thus bringing the applicability of a two dimensional fixed angle grid system for fibre direction determination into question. However, the angulation of the fibres of the multifidus and iliocostalis lumborum were found to be easily identifiable by the use of three surface anatomical landmarks: the caudal tip of the superior iliac spine, the lateral border of the iliocostalis at the twelfth rib and the L1-L2 interspinous space. No reliable index was found for the longissimus. Suggested electrode placement sites for the electromyographic study of the iliocostalis lumborum and the multifidus are at the levels of the L2-L3 and the L4-L5 interspinous spaces respectively.


Tissue regeneration and the epididymal stem cell.
In most pseudostratified epithelia, basal cells represent a multipotent adult stem cell population. These cells generally remain in a quiescent state, until they are stimulated to respond to tissue damage by initiating epithelial regeneration. In the epididymis, cell proliferation occurs at a relatively slow rate under normal physiological conditions. Epididymal basal cells have been shown to share common properties with multipotent adult stem cells. The development of organoids from stem cells represents a novel approach for understanding cellular differentiation and characterization of stem cells. To review the literature on tissue regeneration in the epididymis and demonstrate the presence of an epididymal stem cell population. PubMed database was searched for studies reporting on cell proliferation, regeneration, and stem cells in the epididymis. Three-dimensional cell culture of epididymal cells was used to determine whether these can develop into organoids in a similar fashion to stem cells from other tissues. The epididymal epithelium can rapidly regenerate following orchidectomy or efferent duct ligation, in order to maintain epithelial integrity. Studies have isolated a highly purified fraction of rat epididymal basal cells and reported that these cells displayed properties similar to those of multipotent adult stem cells. In two-dimensional cell culture conditions, these cells differentiated into cells which expressed connexin 26, a marker of columnar cells, and cytokeratin 8. Furthermore, three-dimensional cell culture of epididymal cells resulted in the formation of organoids, a phenomenon associated with the proliferation and differentiation of stem cells in vitro. The rapid proliferation and tissue regeneration of the epididymal epithelium to preserve its integrity following tissue damage as well as the ability of cells to differentiate into organoids in vitro support the notion of a resident progenitor/stem cell population in the adult epididymis.


Metabolic basis of hyperapobetalipoproteinemia. Turnover of apolipoprotein B in low density lipoprotein and its precursors and subfractions compared with normal and familial hypercholesterolemia.
The turnover of apolipoprotein B (apo B) in very low density, intermediate density, and low density lipoproteins (VLDL, IDL, and LDL) and in the light and heavy fractions of LDL was determined in seven patients with hyperapobetalipoproteinemia (hyperapo B), six normolipidemic subjects, and five patients with heterozygous familial hypercholesterolemia (FH). After receiving an injection of 125I-VLDL, hyperapo B patients were found to have a higher rate of synthesis of VLDL-apo B than controls (40.1 vs. 21.5 mg/kg per d, P less than 0.05) but a reduced fractional catabolic rate (FCR) (0.230 vs. 0.366/h, P less than 0.01). After receiving an injection of 131I-LDL, hyperapo B patients had higher rates of LDL-apo B synthesis than controls (23.1 vs. 13.0 mg/kg per d, P less than 0.001), as did FH patients (22.7 mg/kg per d). The FCR of LDL was similar in hyperapo B patients and controls (0.386 vs. 0.366/d) but was markedly decreased in FH patients (0.192/d). Most subjects exhibited precursor-product relationships between VLDL and IDL, and all did between IDL and light LDL; an analogous relationship between light and heavy LDL was evident in most hyperapo B patients and controls but not in FH patients. Simultaneous injection of differentially labeled LDL fractions and deconvolution analysis showed increased light LDL synthesis with normal conversion into heavy LDL in hyperapo B, whereas in FH conversion of light LDL was reduced and there was independent synthesis of heavy LDL. These data show that the increased concentration of LDL-apo B in hyperapo B is solely due to increased LDL synthesis, which is secondary to increased VLDL synthesis; in contrast, in FH there is both an increase in synthesis of LDL (which is partly VLDL-independent) and reduced catabolism.


Cytokines as a therapeutic target for allergic diseases: a complex picture.
Production of IgE antibodies promotes the development of allergic disorders such as asthma, rhinitis and atopic eczema. Though Th2 cytokines play a pivotal role in the allergic inflammatory cascade, therapeutic strategies which target these factors have not been curative in clinical trials. In humans, the allergic phenotype encompasses a broad spectrum of diverse clinical entities, which are dictated by genetics as well as the nature of the allergen and the time in life at which allergen is encountered. The disparate findings in animal and human systems highlight the complexity of cytokine-mediated events in allergic responses in man. Different allergic phenotypes cannot be distinguished strictly on the basis of Th1 and Th2 cytokines. These observations coupled with an emerging role for regulatory T cells in modulation of the allergic response warrant re-examination of the cytokine network in allergic disease. This article highlights the challenges of dissecting the role of individual cytokines in the development of allergic responses and manifestation of allergic symptoms. Regulatory T cells have been implicated in modulation of the activity of allergen-specific Th1 and Th2 effector cells and immune outcome; however, the characteristics of these regulatory T cells remain largely undefined. The effects of existing and emerging therapies which target Th1, Th2 and regulatory cytokines on established allergic responses are examined in the context of this new paradigm. Taken together, existing data suggest that a multi-faceted approach, which is tailored to each patient, will be required in order to attain clinical benefit.


Evaluation of Hybrid Vesicles in an Intestinal Cell Model Based on Structured Paper Chips.
Cell culture-based intestinal models are important to evaluate nanoformulations intended for oral drug delivery. We report the use of a floating structured paper chip as a scaffold for Caco-2 cells and HT29-MTX-E12 cells that are two established cell types used in intestinal cell models. The formation of cell monolayers for both mono- and cocultures in the paper chip are confirmed and the level of formed cell-cell junctions is evaluated. Further, cocultures show first mucus formation between 6-10 days with the mucus becoming more pronounced after 19 days. Hybrid vesicles (HVs) made from phospholipids and the amphiphilic block copolymer poly(cholesteryl methacrylate)-block-poly(2-carboxyethyl acrylate) in different ratios are used as a representative soft nanoparticle to assess their mucopenetration ability in paper chip-based cell cultures. The HV assembly is characterized, and it is illustrated that these HVs cross the mucus layer and are found intracellularly within 3 h when the cells are grown in the paper chips. Taken together, the moist three-dimensional cellulose environment of structured paper chips offers an interesting cell culture-based intestinal model that can be further integrated with fluidic systems or online read-out opportunities.


Intravenous MgSO4 alone and in combination with glucose, insulin and potassium (GIK) prolong the atrial cycle length in chronic atrial fibrillation.
To investigate the effects of parenteral administration of MgSO4, and glucose, insulin, and potassium (GIK), on the dominant atrial cycle length during chronic atrial fibrillation (CAF). The length of the dominant atrial cycle (DACL) in the power-frequency spectrum of the QRST-suppressed lead V1 ECG was identified before and after intravenous administration of MgSO4 alone and after 5 and 10 h of MgSO4 and GIK infusion, in 13 patients with CAF. The changes in DACL were compared with changes in heart rate (HR), blood pressure and blood parameters. MgSO4 alone increased the DACL from 146(13) (mean(SD)) (control) to 153(14) ms (P < 0.01) and decreased the HR from 102(22) to 95(18) beats x min(-1) (P < 0.05). After 5 h of MgSO4 and GIK infusion the DACL was increased compared with control, from 146(13) to 152(11) ms (P < 0.01), but unchanged compared with that after the bolus infusion of MgSO4. HR was decreased compared with control (102(22)) and the bolus infusion of MgSO4 (95(18)) to 87(15) beats x min(-1) after 5 h of intervention. The DACL was further increased after 10 h of MgSO4 and GIK infusion compared with both control (from 146(13) to 157(11) ms), (P < 0.01) and the 5h infusion (152(11) to 157(11) ms), (P < 0.05). No further changes were seen in HR after 10 h (87(17)) of intervention. There were indications of an inverse relationship between total changes in HR (deltaHR) and DACL (deltaDACL) during the interventions (P < 0.05). Bolus infusion of MgSO4 prolongs the DACL and decreases HR in CAF. A further prolongation of DACL was seen after 10 h of MgSO4 and GIK infusion compared with control and with 5 h of intervention. Changes in DACL and HR during the entire intervention period showed an inverse relationship. The antiarrhythmic properties of MgSO4 and the GIK solution in CAF clearly require further attention.


HMOX1-overexpressing mesenchymal stem cell-derived exosomes facilitate diabetic wound healing by promoting angiogenesis and fibroblast function.
Many scholars have suggested that exosomes (Exos) can carry active molecules to induce angiogenesis and thus accelerate diabetic wound healing. Heme oxygenase-1 (HO-1) encoded by the gene HMOX1 promotes wound healing in DM by enhancing angiogenesis. Nevertheless, whether HMOX1 regulates wound healing in DM through mesenchymal stem cell-derived exosomes (MSC-Exos) remains to be further explored. The primary isolated- and cultured-cells expressed MSC-specific marker proteins, and had low immunogenicity and multi-differentiation potential, which means that MSCs were successfully isolated in this study. Notably, HO-1 protein expression was significantly higher in Exo-HMOX1 than in Exos, indicating that HMOX1 could be delivered to Exos as an MSCs-secreted protein. After verifying the -Exo structure, fibroblasts, keratinocytes, and human umbilical vein endothelial cells (HUVECs) were incubated with Exo-HMOX1 or Exo, and the findings displayed that Exo-HMOX1 introduction promoted the proliferation and migration of fibroblasts, keratinocytes and the angiogenic ability of HUVECs in vitro study. After establishing diabetic wound model mice, PBS, Exo, and Exo-HMOX1 were subcutaneously injected into multiple sites on the 1st, 3rd, 7th, and 14th day, DM injected with Exo-HMOX1 showed faster wound healing, re-epithelialization, collagen deposition, and angiogenesis than those in PBS and Exo groups in vitro study. In summary, Exo-HMOX1 could enhance the activity of fibroblasts, keratinocytes, and HUVEC, and accelerate wound healing by promoting angiogenesis in DM.


Quality Control of Herbal Medicines: From Traditional Techniques to State-of-the-art Approaches.
Herbal medicines are important options for the treatment of several illnesses. Although their therapeutic applicability has been demonstrated throughout history, several concerns about their safety and efficacy are raised regularly. Quality control of articles of botanical origin, including plant materials, plant extracts, and herbal medicines, remains a challenge. Traditionally, qualitative (e.g., identification and chromatographic profile) and quantitative (e.g., content analyses) markers are applied for this purpose. The compound-oriented approach may stand alone in some cases (e.g., atropine in Atropa belladonna). However, for most plant materials, plant extracts, and herbal medicines, it is not possible to assure quality based only on the content or presence/absence of one (sometimes randomly selected) compound. In this sense, pattern-oriented approaches have been extensively studied, introducing the use of multivariate data analysis on chromatographic/spectroscopic fingerprints. The use of genetic methods for plant material/plant extract authentication has also been proposed. In this study, traditional approaches are reviewed, although the focus is on the applicability of fingerprints for quality control, highlighting the most used approaches, as well as demonstrating their usefulness. The literature review shows that a pattern-oriented approach may be successfully applied to the quality assessment of articles of botanical origin, while also providing directions for a compound-oriented approach and a rational marker selection. These observations indicate that it may be worth considering to include fingerprints and their data analysis in the regulatory framework for herbal medicines concerning quality control since this is the foundation of the holistic view that these complex products demand.


Stereotactic Navigation Improves Outcomes of Orbital Decompression Surgery for Thyroid Associated Orbitopathy.
To test whether intraoperative stereotactic navigation during orbital decompression surgery resulted in quantifiable surgical benefit. This retrospective cohort study examined all consecutive patients who underwent primary orbital decompression surgery for thyroid associated orbitopathy performed by a single surgeon (A.K.) during the periods of 2012-2014 (non-navigated), and 2017-2018 (navigated). The study was HIPAA-compliant, was approved by the Institutional Review Board, and adhered to the tenets of the Helsinki declaration. Recorded parameters included patient age, sex, race, decompression technique (side of operation and walls decompressed), estimated blood loss (EBL), intraoperative complications, times that patient entered and exited the operating room (OR), times of surgical incision and dressing completion, pre- and postoperative best corrected visual acuity (BCVA), proptosis, diplopia, postoperative change in strabismus deviation, and need for subsequent strabismus surgery. Recorded times were used to calculate operating time (initial incision to dressing) and maintenance time (time between OR entry and initial incision and time between dressings and OR exit). The total maintenance time was averaged over total number of operations. Student t test was used to compare surgical times, maintenance times, EBL, and proptosis reduction. Fisher exact test was used to compare BCVA change, strabismus deviation change, resolution or onset of diplopia, and need for corrective strabismus surgery. Twenty-two patients underwent primary orbital decompression surgery without navigation, and 23 patients underwent navigation-guided primary orbital decompression surgery. There were no intraoperative complications in either group. The average operative time was shorter in the navigated group for a unilateral balanced decompression (n = 10 vs. 19; 125.8 ± 13.6 vs. 141.3 ± 19.4 min; p-value = 0.019), and a unilateral lateral wall only decompression (n = 13 vs. 3; 80.5 ± 12.8 vs. 93.0 ± 6.1 min; p-value = 0.041). The average maintenance time per surgery was not significantly different between the non-navigated group (51.3 ± 12.7 min) and the navigated group (50.5 ± 6.4 min). There was no significant difference between the navigated and non-navigated groups in average EBL per surgery. There was no significant difference in BCVA change. Average proptosis reduction was larger in the navigated group, but this was not significant. There was a significantly lower proportion of patients who required corrective strabismus surgery following decompression in the navigated group than in the non-navigated group (39.1% vs. 77.3%, p-value = 0.012). Intraoperative stereotactic navigation during orbital decompression surgery has the potential to provide the surgeon with superior spatial awareness to improve patient outcomes. This study found that use of intraoperative navigation reduced operative time (even without factoring in a resident teaching component) while also reducing the need for subsequent strabismus surgery. This study is limited by its size but illustrates that use of intraoperative navigation guidance has substantive benefits in orbital decompression surgery.


The response of red cell hexose monophosphate shunt after sulfhydryl inhibition.
In this investigation, we studied the importance of cellular glutathione (GSH) in the hexose monophosphate shunt (HMPS) activity of unstimulated human erythrocytes and the mechanism by which pyruvate stimulates the HMPS. The rate of HMPS activity was measured by the production of radioactive CO2 from 14C-1-glucose or 14C-1-ribose using a vibrating reed electrometer and ionization chamber. HMPS activity was not significantly impaired by N-ethylmaleimide (NEM) in concentrations which bound all red cell GSH. Red cells incubated under carbon monoxide (CO), an experimental condition which eliminates peroxide production, still had HMPS activity which was 44% of the value under air. Pyruvate stimulation of the HMPS was unaffected by doses of NEM which bound all cellular GSH or by incubation under CO. These data indicated that pyruvate stimulation of the HMPS occurs by pathways which do not involve peroxide formation, GSH, or oxygen. This study indicates that sulfhydryl blockade of GSH does not necessarily inhibit HMPS activity and that HMPS activity in red cells may respond to reactions not linked directly to glutathione reduction.


Viral activation of macrophages through TLR-dependent and -independent pathways.
Induction of cytokine production is important for activation of an efficient host defense response. Macrophages constitute an important source of cytokines. In this study we have investigated the virus-cell interactions triggering induction of cytokine expression in macrophages during viral infections. We found that viral entry and viral gene products produced inside the cell are responsible for activation of induction pathways leading to IFN-alphabeta expression, indicating that virus-cell interactions on the cell surface are not enough. Moreover, by the use of cell lines expressing dominant negative versions of TLR-associated adaptor proteins we demonstrate that Toll/IL-1 receptor domain-containing adaptor inducing IFN-beta is dispensable for all virus-induced cytokine expression examined. However, a cell line expressing dominant negative MyD88 revealed the existence of distinct induction pathways because virus-induced expression of RANTES and TNF-alpha was totally blocked in this cell line whereas IFN-alphabeta expression was much less affected in the absence of signaling via MyD88. In support of this, we also found that inhibitory CpG motifs, which block TLR9 signaling inhibited early HSV-2-induced TNF-alpha and RANTES expression dramatically whereas IFN-alphabeta induction was only slightly affected. This suggests that virus activates macrophages through distinct pathways, of which some are dependent on TLRs signaling through MyD88, whereas others seem to be independent of TLR signaling. Finally we demonstrate that IFN-alphabeta induction in HSV-2-infected macrophages requires a functional dsRNA-activated protein kinase molecule because cells expressing a dsRNA-dependent protein kinase version unable to bind dsRNA do not express IFN-alphabeta on infection.


Predictors of Anastomotic Leak in Elderly Patients After Colectomy: Nomogram-Based Assessment From the American College of Surgeons National Surgical Quality Program Procedure-Targeted Cohort.
Elderly patients undergoing colorectal surgery have increasingly become under scrutiny by accounting for the largest fraction of geriatric postoperative deaths and a significant proportion of all postoperative complications, including anastomotic leak. This study aimed to determine predictors of anastomotic leak in elderly patients undergoing colectomy by creating a novel nomogram for simplistic prediction of anastomotic leak risk in a given patient. This study was a retrospective review. The database review of the American College of Surgeons National Surgical Quality Improvement Program was conducted at a single institution. Patients aged ≥65 years who underwent elective segmental colectomy with an anastomosis at different levels (abdominal or low pelvic) in 2012-2013 were identified from the multi-institutional procedure-targeted database. We constructed a stepwise multiple logistic regression model for anastomotic leak as an outcome; predictors were selected in a stepwise fashion using the Akaike information criterion. The validity of the nomogram was externally tested on elderly patients (≥65 years of age) from the 2014 American College of Surgeons National Surgical Quality Improvement Program colectomy-targeted database. A total of 10,392 patients were analyzed, and anastomotic leak occurred in 332 (3.2%). Of the patients who developed anastomotic leak, 192 (57.8%) were men (p < 0.001). Based on unadjusted analysis, factors associated with an increased risk of anastomotic leak were ASA score III and IV (p < 0.001), chronic obstructive pulmonary disease (p = 0.004), diabetes mellitus (p = 0.003), smoking history (p = 0.014), weight loss (p = 0.013), previously infected wound (p = 0.005), omitting mechanical bowel preparation (p = 0.005) and/or preoperative oral antibiotic use (p < 0.001), and wounds classified as contaminated or dirty/infected (p = 0.008). Patients who developed anastomotic leak had a longer length of hospital stay (17 vs 7 d; p < 0.001) and operative time (191 vs 162 min; p < 0.001). A multivariate model and nomogram were created. This study was limited by its retrospective nature and short-term follow-up (30 d). An accurate prediction of anastomotic leak affecting morbidity and mortality after colorectal surgery using the proposed nomogram may facilitate decision making in elderly patients for healthcare providers.


Identification and analysis of cabut orthologs in invertebrates and vertebrates.
Cabut (cbt) is a Drosophila melanogaster gene involved in epidermal dorsal closure (DC). Its expression is dependent on the Jun N-terminal kinase (JNK) cascade, and it functions downstream of Jun regulating dpp expression in the leading edge cells. The Cbt protein contains three C(2)H(2)-type zinc fingers and a serine-rich domain, suggesting that it functions as a transcription factor. We have identified single cbt orthologs in other Drosophila species, as well as in other insects and invertebrate organisms like ascidians and echinoderms, but not in nematodes. Gene structure and protein sequence are highly conserved among Drosophilidae, but are more diverged in the other species of invertebrates analyzed. According to this, we demonstrate that cbt expression is detected in the embryonic lateral epidermis in several Drosophila species, as it occurs in D. melanogaster, thus suggesting that the cbt orthologs may have a conserved role in these species during DC. We have also analyzed the genomes of several vertebrate species, finding that the cbt orthologous genes in these organisms encode proteins that belong to the TIEG family of Sp1-like/Krüppel-like transcription factors. Phylogenetic analysis of the invertebrate and vertebrate proteins identified indicates that they mainly follow the expected phylogeny of the species, and that the cbt gene was duplicated during vertebrate evolution. Because we were not able to identify cbt orthologous genes neither in yeast nor in plants, our results suggest that this gene has been probably conserved throughout metazoans and that it may play a fundamental role in animal biology.


Synergistic effect of up-flow constructed wetland and microbial fuel cell for simultaneous wastewater treatment and energy recovery.
This study demonstrated a successful operation of up-flow constructed wetland-microbial fuel cell (UFCW-MFC) in wastewater treatment and energy recovery. The goals of this study were to investigate the effect of circuit connection, organic loading rates, and electrode spacing on the performance of wastewater treatment and bioelectricity generation. The average influent of COD, NO3(-) and NH4(+) were 624 mg/L, 142 mg/L, 40 mg/L, respectively and their removal efficiencies (1 day HRT) were 99%, 46%, and 96%, respectively. NO3(-) removal was relatively higher in the closed circuit system due to lower dissolved oxygen in the system. Despite larger electrode spacing, the voltage outputs from Anode 2 (A2) (30 cm) and Anode 3 (A3) (45 cm) were higher than from Anode 1 (A1) (15 cm) as a result of insufficient fuel supply to A1. The maximum power density and Coulombic efficiency were obtained at A2, which were 93 mW/m(3) and 1.42%, respectively.


Radiologic evaluation of the solitary pulmonary nodule.
The solitary pulmonary nodule (SPN) is a common radiologic finding. There are different management approaches and the work-up often requires evaluation over a long period of time to establish a benign or malignant diagnosis. Comparison with old examinations and morphologic evaluation of the size, margins, and internal characteristics of an (SPN) should be the first step in the evaluation of these lesions. It is often necessary to proceed to additional imaging techniques and in some situations invasive tests or surgical biopsy may be required.


The UNOS Scientific Renal Transplant Registry.
1. One-year graft survival rates for recipients of cadaver kidney transplants improved from 75% in 1988 to 83% in 1991 (p < 0.001). The one-, 5-, and projected 10-year graft survival rates for cadaver donor transplants performed in 1991-1995 were 84%, 60%, and 43% respectively. 2. One-year graft survival rates for recipients of living donor transplants also improved from 89% in 1988 to 93% in 1991 (p < 0.001). The one-, 5-, and projected 10-year graft survival rates for living donor kidney transplants performed in 1991-1995 were 92%, 75%, and 62% respectively. 3. Diabetic patients received one-quarter of the cadaver kidneys transplanted from 1991-1995 and one third of diabetic patients received a simultaneous pancreas (SPK) transplant. One- and 5-year graft survival rates were 81% and 54% for diabetics receiving a kidney transplant and 85% and 67% for SPK recipients, respectively. Patient survival was 10% lower for recipients of a kidney only transplant. 4. Sensitization to alloantigens, whether by pregnancy, transfusion, or graft failure resulted in about a 5% increased risk of early graft failure. Patients who developed broadly reactive anti-HLA antibodies before their first transplant had the same 5-year graft survival rate (60%) as unsensitized patients. Retransplanted patients who had not developed broadly reactive antibodies also had a 60% 5-year graft survival rate, compared with 50% for those with > 50% PRA. 5. Blacks received one-quarter of cadaver kidneys transplanted in 1991-1995. The one-year graft survival rate for Black first transplant recipients was 83% compared with 84% for Whites. After the first year, the graft loss rate among Blacks was almost double that for other racial groups (5.8 year half-life vs 11.3 years for Whites, p < 0.01). The 5-year graft survival rate was 49% among Blacks and 63% for Whites. Asian recipients had the highest one- and 5-year graft survival rates (89% and 70%, respectively). 6. Shared kidneys had a longer average cold ischemia time (30 hr) than kidneys transplanted locally (21 hr). Fewer than half of shared kidneys were transplanted to HLA-matched recipients. The 5-year graft survival rate for shared kidneys with zero or one HLA antigen mismatched was 68% compared with 59% for shared kidneys with more than 3 antigens mismatched and for locally transplanted kidneys (p < 0.001). 7. The distribution of living donor relationships has changed substantially. When comparing transplants performed in 1988-1989 with those performed in 1994-1995, the number of living donor transplants increased by 80%, the fraction of offspring-to-parent grafts increased from 9-15%, the fraction of genetically unrelated donors increased from 4-10%, and the fraction of distant relatives increased from 2-6% of the living donor transplants. 8. The results of living donor transplants generally followed the degree of HLA compatibility. The one-year survival rate for HLA-identical sibling grafts was 96%, followed by 92% for one-haplotype matched sibling, parent and offspring donor transplants, 90% for unmatched sibling donors and 88% for spousal donors. Other unrelated donor transplants had a slightly higher one-year graft survival rate of 92%, which was more similar to the one-haplotype matched grafts.


Evaluation of blind thoracoscopic-assisted placement of three double-lumen endobronchial tube designs for one-lung ventilation in dogs.
To describe a technique for blind thoracoscopic-assisted double-lumen endobronchial tube (DLT) placement for achieving one-lung ventilation (OLV) using 3 different DLT designs and to evaluate whether thoracic auscultation could reliably confirm OLV in dogs. Prospective randomized study. Mature female hound dogs (n = 6). Physical examination, baseline blood work and thoracic radiography confirmed the absence of preexisting disease in all dogs. Thoracoscopic observation was established through a subxiphoid portal and used for evaluation of differential lung lobe ventilation. Each dog was sequentially intubated using 1 of 3 DLT designs in random order; Robertshaw left-sided tube (RS-L), Carlens left-sided tube (C-L), Dr. White right-sided tube (DW-R). Incidence of initial and overall (after a maximum of 3 DLT manipulations) correct and complete OLV (CC-OLV) was recorded. After each blind thoracoscopic-assisted DLT placement, bronchoscopic evaluation was performed to document correct DLT position. Blind DLT placement achieved overall CC-OLV in all dogs using RS-L, 66% using C-L, and all using DW-R. Successful initial left-sided OLV (L-OLV) was statistically more likely when RS-L DLTs were used compared to C-L or DW-R. Of cases where overall CC-OLV was achieved, correct DLT position was present in only 44% of intubations. Blind DLT placement produced successful CC-OLV in all dogs using at least one DLT design evaluated. Use of thoracoscopic assistance may obviate the need for bronchoscopic observation during DLT placement in dogs. However, bronchoscopic observation may still refine DLT positioning in dogs after blind thoracoscopic-assisted DLT placement.


Telephone Coaching to Enhance a Home-Based Physical Activity Program for Knee Osteoarthritis: A Randomized Clinical Trial.
To investigate whether simultaneous telephone coaching improves the clinical effectiveness of a physiotherapist-prescribed home-based physical activity program for knee osteoarthritis (OA). A total of 168 inactive adults ages ≥50 years with knee pain on a numeric rating scale ≥4 (NRS; range 0-10) and knee OA were recruited from the community and randomly assigned to a physiotherapy (PT) and coaching group (n = 84) or PT-only (n = 84) group. All participants received five 30-minute consultations with a physiotherapist over 6 months for education, home exercise, and physical activity advice. PT+coaching participants also received 6-12 telephone coaching sessions by clinicians trained in behavioral-change support for exercise and physical activity. Primary outcomes were pain (NRS) and physical function (Western Ontario and McMaster Universities Osteoarthritis Index [WOMAC; score range 0-68]) at 6 months. Secondary outcomes were these same measures at 12 and 18 months, as well as physical activity, exercise adherence, other pain and function measures, and quality of life. Analyses were intent-to-treat with multiple imputation for missing data. A total of 142 (85%), 136 (81%), and 128 (76%) participants completed 6-, 12-, and 18-month measurements, respectively. The change in NRS pain (mean difference 0.4 unit [95% confidence interval (95% CI) -0.4, 1.3]) and in WOMAC function (1.8 [95% CI -1.9, 5.5]) did not differ between groups at 6 months, with both groups showing clinically relevant improvements. Some secondary outcomes related to physical activity and exercise behavior favored PT+coaching at 6 months but generally not at 12 or 18 months. There were no between-group differences in most other outcomes. The addition of simultaneous telephone coaching did not augment the pain and function benefits of a physiotherapist-prescribed home-based physical activity program.


Dental mesenchymal stem cells encapsulated in an alginate hydrogel co-delivery microencapsulation system for cartilage regeneration.
Dental-derived mesenchymal stem cells (MSCs) are promising candidates for cartilage regeneration, with a high capacity for chondrogenic differentiation. This property helps make dental MSCs an advantageous therapeutic option compared to current treatment modalities. The MSC delivery vehicle is the principal determinant for the success of MSC-mediated cartilage regeneration therapies. The objectives of this study were to: (1) develop a novel co-delivery system based on TGF-β1 loaded RGD-coupled alginate microspheres encapsulating periodontal ligament stem cells (PDLSCs) or gingival mesenchymal stem cells (GMSCs); and (2) investigate dental MSC viability and chondrogenic differentiation in alginate microspheres. The results revealed the sustained release of TGF-β1 from the alginate microspheres. After 4 weeks of chondrogenic differentiation in vitro, PDLSCs and GMSCs as well as human bone marrow mesenchymal stem cells (hBMMSCs) (as positive control) revealed chondrogenic gene expression markers (Col II and Sox-9) via qPCR, as well as matrix positively stained by Toluidine Blue and Safranin-O. In animal studies, ectopic cartilage tissue regeneration was observed inside and around the transplanted microspheres, confirmed by histochemical and immunofluorescent staining. Interestingly, PDLSCs showed more chondrogenesis than GMSCs and hBMMSCs (p<0.05). Taken together, these results suggest that RGD-modified alginate microencapsulating dental MSCs make a promising candidate for cartilage regeneration. Our results highlight the vital role played by the microenvironment, as well as value of presenting inductive signals for viability and differentiation of MSCs.


D-Amphetamine Exposure Differentially Disrupts Signaling Across Ontogeny in the Zebrafish.
Background: Prescriptive and illicit amphetamine (AMPH) use continues to increase along with the likelihood that during an individual's lifetime, the drug deleteriously influences the growth and connectivity of behavior circuits necessary for survival. Throughout ontogeny, neural circuits underlying these behaviors grow in complexity, gradually integrating many sensory inputs that trigger higher order coordinated motor responses. In the present study, we examine how AMPH disrupts the establishment of these circuits at critical neurodevelopmental periods, as well as the communication among established survival circuits. Materials and Methods: Zebrafish embryos (from 1 hpf) were raised in AMPH solutions, growth parameters and escape behavior were assessed at 24 and 48 hpf, and spinal cord tissues analyzed for differences in excitatory-inhibitory signaling balance among treatments. Adult fish were fed an acute dosage of AMPH over an 11-day conditioned place preference (PP) paradigm during which behaviors were recorded and brain tissues analyzed for alterations in dopaminergic signaling. Results: AMPH negatively affects embryonic growth and slows the execution of escape behavior, suggesting an imbalance in locomotor signaling. Although local spinal circuits provide primary escape modulation, no differences in inhibitory glycinergic, and excitatory glutamatergic signaling were measured among spinal neurons. AMPH also influenced place preference in adult zebrafish and resulted in the increased expression of dopamine signaling proteins (DRD1) in brain areas governing survival behaviors.


Peri-Operative Intracameral Antibiotics: The Perfect Storm?
In our University journal club we discussed a large, retrospective study of cataract surgery endophthalmitis rates before and after instituting the use of an intracameral fluoroquinolone antibiotic. We identified several factors involved in the use of off-label, compounded moxifloxacin in intraocular surgery. The introduction of phacoemulsification for cataract surgery led to the potential for smaller incisions. Intraocular lens technology improved to allow for foldable lenses, obviating the requirement to enlarge the incision. This allowed for clear corneal incisions, which unfortunately allow bidirectional passage of liquid. Preservatives were introduced into multi-dose ophthalmic products in the mid 20th century to retard microbial growth. However, more recently, chronic use of benzalkonium chloride has led to concerns about concerns about conjunctival toxicity, especially in patients with ocular surface disease. In the formulation of ocular moxifloxacin, developers were able to develop a "self-preserved", multi-dose product. Other concerns with eyedrops include varying levels of adherence and performance, and the expansion of compounding pharmacies from a named-patient basis to widespread national delivery, with concerns for lower quality. Integrating these factors, use of intracameral moxifloxacin as a prophylactic during cataract and other anterior segment surgery has become a standard of care in much of the U.S. We are concerned that the current position is on a narrow ledge-the standard of care for millions of surgeries each year based upon off-label, compounting use of a single product. We discuss possible ramifications and solutions to this public health issue.


Promethazine inhibits the formation of aldehydic products of lipid peroxidation but not covalent binding resulting from the exposure of rat liver fractions to CCl4.
Promethazine is known to have protective activity in relation to CCl4-induced liver necrosis. This hepatoprotective property has been investigated with regard to the free radical scavenging and antioxidant properties of promethazine using isolated hepatocytes and microsomal suspensions. CCl4 is activated in both systems to free radical metabolites that bind covalently to lipid and protein, and initiate lipid peroxidation. A large number of carbonyl products is produced during CCl4-induced lipid peroxidation; promethazine strongly inhibits the production of all classes of carbonyl compounds in both microsomal suspensions and isolated hepatocytes. In contrast, promethazine is a very weak inhibitor of the covalent binding of metabolites of CCl4. We conclude that promethazine acts by scavenging the trichloromethylperoxyl radical and lipid peroxyl radicals, and is a weak scavenger of the trichloromethyl radical. These data, when considered together with the hepatoprotective effects of promethazine, suggest that lipid peroxidation is of relatively more importance than covalent binding in the pathogenesis of CCl4-induced liver necrosis.


Executive functions in trauma-exposed youth: a meta-analysis.
An earlier meta-analysis and review indicated that trauma exposure may be related to lower levels of executive functioning in youth. Since different developmental trajectories were found for three core executive functions, the present study focused on working memory, inhibition, and cognitive flexibility specifically. We conducted a multi-level meta-analysis on 55 studies and 322 effect sizes published between 2001 and 2017 that were retrieved from MEDLINE, Embase, and PsycINFO. The 8070 participants in selected studies were aged 2-25 years. We investigated whether the association between constructs would be moderated by trauma-specific moderators (onset, duration, and type), and study (age, gender, ethnicity, and socio-economic status) and measurement (quality) characteristics. We found small to medium effect sizes for working memory (d = -0.49), inhibition (d = -0.46), and cognitive flexibility (d = -0.44). Moderator analyses showed that, for working memory, when studies used low-quality measurements the effect size was significantly stronger than when studies used high-quality measurements.Compared to single trauma-exposed youth, violence-exposed/abused and foster care/adopted youth showed more problems in inhibition, and foster care/adopted youth showed more problems in cognitive flexibility. Our findings imply that trauma-exposed youth have lower levels of executive functions. Clinical practice should incorporate problems in executive functioning, especially working memory, inhibition, and cognitive flexibility, in assessment and treatment guidelines.


Digital technologies to support adolescents with depression and anxiety: review.
Depression and anxiety are common in adolescents, but most affected will not get any formal help. Digital mental health technologies (i.e. resources and interventions to support and improve mental health) are a potential way to extend the reach and increase adolescents' access to therapies, at a relatively low cost. Many young people can access the internet and mobile technologies, including in low- and middle-income countries. There has been increased interest in integrating technologies in a range of settings, especially because of the effect of the COVID-19 pandemic on adolescent mental health, at a time when services are under pressure. This clinical review gives an overview of digital technologies to support the prevention and management of depression and anxiety in adolescence. The technologies are presented in relation to their technological approaches, underlying psychological or other theories, setting, development, evaluations to date and how they might be accessed. There is also a discussion of the potential benefits, challenges and future developments in this field.


Active Brownian equation of state: metastability and phase coexistence.
As a result of the competition between self-propulsion and excluded volume interactions, purely repulsive self-propelled spherical particles undergo a motility-induced phase separation (MIPS). We carry out a systematic computational study, considering several interaction potentials, systems confined by hard walls or with periodic boundary conditions, and different initial conditions. This approach allows us to identify that, despite its non-equilibrium nature, the equations of state of Active Brownian Particles (ABP) across MIPS verify the characteristic properties of first-order liquid-gas phase transitions, meaning, equality of pressure of the coexisting phases once a nucleation barrier has been overcome and, in the opposite case, hysteresis around the transition as long as the system remains in the metastable region. Our results show that the equations of state of ABPs account for their phase behaviour, providing a firm basis to describe MIPS as an equilibrium-like phase transition.


Characterization of an orthovoltage biological irradiator used for radiobiological research.
Orthovoltage irradiators are routinely used to irradiate specimens and small animals in biological research. There are several reports on the characteristics of these units for small field irradiations. However, there is limited knowledge about use of these units for large fields, which are essential for emerging large-field irregular shape irradiations, namely total marrow irradiation used as a conditioning regimen for hematological malignancies. This work describes characterization of a self-contained Orthovoltage biological irradiator for large fields using measurements and Monte Carlo simulations that could be used to compute the dose for in vivo or in vitro studies for large-field irradiation using this or a similar unit. Percentage depth dose, profiles, scatter factors, and half-value layers were measured and analyzed. A Monte Carlo model of the unit was created and used to generate depth dose and profiles, as well as scatter factors. An ion chamber array was also used for profile measurements of flatness and symmetry. The output was determined according to AAPM Task Group 61 guidelines. The depth dose measurements compare well with published data for similar beams. The Monte Carlo-generated depth dose and profiles match our measured doses to within 2%. Scatter factor measurements indicate gradual variation of these factors with field size. Dose rate measured by placing the ion chamber atop the unit's steel plate or solid water indicate enhanced readings of 5 to 28% compared with those measured in air. The stability of output over a 5-year period is within 2% of the 5-year average.


Content of low-molecular-weight thiols during the imbibition of Pea seeds.
The metabolism of low-molecular-weight thiols was investigated in seeds of Pisum sativum L. cv. Kleine Rheinländerin during imbibition in water for 14 h. The amount of oxidized glutathione (GSSG) decreased from 319 nmol (g dry weight)-1 in dry seeds to 38 nmol (g dry weight)-1 within the first 14 h of imbibition. The decrease may have been due to the reduction of GSSG to reduced glutathione (GSH), catalyzed by the enzyme glutathione reductase (GR; EC 220.127.116.11). The enzyme activity was high in dry seeds [25 nkat (g dry weight)-1 ] and decreased to 20 nkat (g dry weight)-1 within 14 h of imbibition. The activity of glucose-6-phosphate dehydrogenase (EC 220.127.116.11) decreased from 100 nkat (g dry weight)-1 in dry seeds to 67 nkat (g dry weight)-1 after 14 h of imbibition. Within 14 h the amount of γ-glutamyl-cysteine (γ-GC) decreased from 135 to 38 nmol (g dry weight)-1 , whereas the cysteine content rose from 81 nmol (g dry weight)-1 in dry seeds to a maximum of 170 nmol (g dry weight)-1 after 12 h of imbibition, which may be due to the degradation of γ-GC into cysteine.


The activities of lipases and carnitine palmitoyltransferase in muscles from vertebrates and invertebrates.
1. The activities of tri-, di- and mono-glyceride lipase and carnitine palmitoyltransferase were measured in homogenates of a variety of muscles. These activities were used to estimate the rate of utilization of glycerides and fatty acids by muscle. In muscles whose estimated rates of fat utilization can be compared with rates calculated for the intact muscle from such information as O(2) uptake, there is reasonable agreement between the estimated and calculated rates. 2. In all muscles investigated the maximum rates of hydrolysis of glycerides increase in the order triglyceride, diglyceride, monoglyceride. The activity of diglyceride lipase is highest in the flight muscles of insects such as the locust, waterbug and some moths and is lowest in the flight muscles of flies, bees and the wasp. These results are consistent with the utilization of diglyceride as a fuel for some insect flight muscles. 3. In many muscles from both vertebrates and invertebrates the activity of glycerol kinase is similar to that of lipase. It is concluded that in these muscles the metabolic role of glycerol kinase is the removal of glycerol produced during lipolysis. However, in some insect flight muscles the activity of glycerol kinase is much greater than that of lipase, which suggests a different role for glycerol kinase in these muscles.


Cardiac myosin activation part 1: from concept to clinic.
Decreased cardiac contractility is a central feature of systolic heart failure and yet we have no effective drugs to improve cardiac contractility. Existing drugs that increase cardiac contractility do so indirectly through signaling cascades and their use is limited by their mechanism-related adverse effects. Direct activation of the cardiac sarcomere to increase cardiac contractility may provide a means to avoid these limitations. Using a reconstituted version of the cardiac sarcomere, we screened a small molecule library and identified several chemical classes that directly activate cardiac myosin. One compound class has been optimized extensively using an iterative process; omecamtiv mecarbil, a small-molecule, selective, cardiac myosin activator is the most advanced exemplar of this novel mechanistic class. It accelerates the transition of myosin into the force-generating state without affecting cardiac myocyte calcium homeostasis. In animal models, omecamtiv mecarbil increases cardiac function by increasing the duration of ejection without changing the rates of contraction. Initial clinical studies have demonstrated the translation of this mechanism into humans, and further clinical studies of its use in acute and chronic heart failure are planned. Cardiac myosin activation may provide a new therapeutic approach for systolic heart failure. This article is part of a special issue entitled "Key Signaling Molecules in Hypertrophy and Heart Failure."


A single-centre chart review exploring the adjusted association between breast cancer phenotype and prognosis.
Using a retrospective chart review, we investigated the differences in survival and prognostic factors between patients with triple-negative breast cancer (tnbc) and those with non-tnbc. The review included 1018 breast cancer patients who were diagnosed between 2000 and 2005 in Essex, Kent, and Lambton counties in Ontario, Canada. Our findings indicate that, although the unadjusted results suggested that patients with tnbc were more likely than patients with non-tnbc to die [hazard ratio (hr): 2.29; 95% confidence interval (ci): 1.33 to 2.93], an adjusted survival analysis revealed no significant difference in overall survival between the groups (hr: 1.22; 95% ci: 0.63 to 2.39). The significant predictors of survival in the adjusted analysis were age, stage of cancer, and size of cancer. Our findings support those of earlier reports, which suggest that presenting tumour size is the most important prognostic factor in tnbc. Investigations into unique screening methods to identify these tumours at an earlier stage and to prevent advanced-stage cancer in this patient subpopulation are necessary.


Single-nucleotide polymorphisms of methylenetetrahydrofolate reductase gene in a South Indian cohort with nonsyndromic cleft lip with or without palate.
Clefts of the lip, with or without cleft palate and cleft palate only, collectively called as orofacial clefts (OFCs) are one of the most common congenital malformations with varying degrees of penetrance and phenotype expressions. The aim of this study was to investigate the association between methylenetetrahydrofolate reductase (MTHFR) cytosine-to-thymine (c. 677 C>T), adenine-to-cytosine (c.1298 A>C) single- nucleotide polymorphisms (SNPs) and South Indian patients with the nonsyndromic cleft lip with or without palate (NSCL ± P). A cohort consisting of 25 cases of NSCL ± P and 18 controls from a South Indian cohort were included in this case-control study. Genetic analysis of c.677C>T and c.1298A C polymorphisms in the MTHFR gene was carried out using Sanger sequencing and analyzed from chromatogram profiles. Data interpretation was done using statistical software MedCalc Statistical Software version 16.2 and the Statistical Package for the Social Sciences (SPSS version 22.0). DNA sequence analysis of the MTHFR gene revealed c. 677C>T and c. 1298A>C polymorphisms in 16% and 76% of NSCL ± P cases, respectively. Heterozygous variant in MTHFR c. 1298A>C polymorphism was found to be a significant risk factor (P = 0.0164) for NSCL ± P in South Indian ethnic population. c.677C>T polymorphism, in particular, was apparently dormant overall in the study population. These results offer certain novelty in terms of the distinctive pattern in SNPs of genotypes observed in the study. NSCL ± P is one of the most common and challenging congenital malformations with complex etiological basis. Common risk factors such as MTHFR SNPs, namely c.677C>T and c.1298A>C, are subjected to variations in terms of ethnic group, geographic region and micro/macro-environmental factors. Overall, our study has explored part of South Indian ethnic population and revealed a different and unique distribution of mutations in this sample population.


Apportioning riverine DIN load to export coefficients of land uses in an urbanized watershed.
The apportionment of riverine dissolved inorganic nitrogen (DIN) load to individual land use on a watershed scale demands the support of accurate DIN load estimation and differentiation of point and non-point sources, but both of them are rarely quantitatively determined in small montane watersheds. We introduced the Danshui River watershed of Taiwan, a mountainous urbanized watershed, to determine the export coefficients via a reverse Monte Carlo approach from riverine DIN load. The results showed that the dynamics of N fluctuation determines the load estimation method and sampling frequency. On a monthly sampling frequency basis, the average load estimation of the methods (GM, FW, and LI) outperformed that of individual method. Export coefficient analysis showed that the forest DIN yield of 521.5kg-Nkm(-2)yr(-1) was ~2.7-fold higher than the global riverine DIN yield (mainly from temperate large rivers with various land use compositions). Such a high yield was attributable to high rainfall and atmospheric N deposition. The export coefficient of agriculture was disproportionately larger than forest suggesting that a small replacement of forest to agriculture could lead to considerable change of DIN load. The analysis of differentiation between point and non-point sources showed that the untreated wastewater (non-point source), accounting for ~93% of the total human-associated wastewater, resulted in a high export coefficient of urban. The inclusion of the treated and untreated wastewater completes the N budget of wastewater. The export coefficient approach serves well to assess the riverine DIN load and to improve the understanding of N cascade.


Castor bean allergens: evidence for distinct heat-labile and stable entities.
Castor beans contain one of the most potent allergens known to man. Previous studies suggested that these allergens are heat-stable; but our current data suggest that both heat-stable and labile allergens are present in castor bean extracts. The heat-labile allergen is of large molecular weight and is immunochemically distinct from the smaller molecular weight heat-stable allergen. Both allergens have different isoelectric points. These antigens may be clinically relevant since coffee workers sensitized to castor have IgE antibodies to both allergens.


Development of machine learning models for diagnosis of glaucoma.
The study aimed to develop machine learning models that have strong prediction power and interpretability for diagnosis of glaucoma based on retinal nerve fiber layer (RNFL) thickness and visual field (VF). We collected various candidate features from the examination of retinal nerve fiber layer (RNFL) thickness and visual field (VF). We also developed synthesized features from original features. We then selected the best features proper for classification (diagnosis) through feature evaluation. We used 100 cases of data as a test dataset and 399 cases of data as a training and validation dataset. To develop the glaucoma prediction model, we considered four machine learning algorithms: C5.0, random forest (RF), support vector machine (SVM), and k-nearest neighbor (KNN). We repeatedly composed a learning model using the training dataset and evaluated it by using the validation dataset. Finally, we got the best learning model that produces the highest validation accuracy. We analyzed quality of the models using several measures. The random forest model shows best performance and C5.0, SVM, and KNN models show similar accuracy. In the random forest model, the classification accuracy is 0.98, sensitivity is 0.983, specificity is 0.975, and AUC is 0.979. The developed prediction models show high accuracy, sensitivity, specificity, and AUC in classifying among glaucoma and healthy eyes. It will be used for predicting glaucoma against unknown examination records. Clinicians may reference the prediction results and be able to make better decisions. We may combine multiple learning models to increase prediction accuracy. The C5.0 model includes decision rules for prediction. It can be used to explain the reasons for specific predictions.


No evidence of geographical structure of salicinoid chemotypes within Populus tremula.
Salicinoids are well-known defense compounds in salicaceous trees and careful screening at the population level is warranted to fully understand their diversity and function. European aspen, Populus tremula, is a foundation species in Eurasia and highly polymorphic in Sweden. We exhaustively surveyed 102 replicated genotypes from the Swedish Aspen collection (SwAsp) for foliar salicinoids using UHPLC-ESI-TOF/MS and identified nine novel compounds, bringing the total to 19 for this species. Salicinoid structure followed a modular architecture of a salicin skeleton with added side groups, alone or in combination. Two main moieties, 2'-cinnamoyl and 2'-acetyl, grouped the SwAsp population into four distinct chemotypes, and the relative allocation of salicinoids was remarkably constant between different environments, implying a highly channeled biosynthesis of these compounds. Slightly more than half of the SwAsp genotypes belonged to the cinnamoyl chemotype. A fraction synthesized the acetyl moiety alone (∼7%) or in combination with cinnamoyl (∼2%), and close to forty percent lacked either of the two characteristic moieties, and thus resemble P. tremuloides in their salicinoid profile. The two most abundant chemotypes were evenly distributed throughout Sweden, unlike geographical patterns reported for SwAsp phenology traits, plant defense genes, and herbivore community associations. Here we present the salicinoid characterization of the SwAsp collection as a resource for future studies of aspen chemical ecology, salicinoid biosynthesis, and genetics.


Prevalence of cardiac arrhythmias in cannabis use disorder related hospitalizations in teenagers from 2003 to 2016 in the United States.
Cannabis is an increasingly common recreational substance used by teenagers. However, there is limited data probing association of cardiac arrhythmias with marijuana use in this population. We provide prevalence trends, disease burden and healthcare utilization of cardiac arrhythmias associated with cannabis use disorder (CUD) in hospitalized teenagers (13-20 years) using a large national administrative database in the United States from 2003-2016. We used partial least square regression analysis for assessing trends in prevalence of cardiac arrhythmias and multiple logistic regression to elucidate independent predictors of arrhythmias associated with CUD. Among all CUD related hospitalizations (n = 876, 431), 0.5% had arrhythmias. Prevalence trends of arrhythmias among CUD increased six-fold during the study period (P < 0.001). CUD was more prevalent in males and older teens (both P < 0.001). There was a significant risk for mortality when CUD was associated with arrhythmia (7.4% vs. 0.1%, P < 0.001). While mean length-of-stay (LOS) was shorter (4.4 vs. 5.4 days, P < 0.001) for patients with CUD, they incurred three times higher mean hospitalization charges when compared to CUD patients without arrhythmia ($45 959 vs. $18 986, P < 0.001). Both LOS and hospitalization charges showed an uptrend during the study period (P < 0.001). Congenital heart disease, congestive heart failure, hypertension, and obesity independently predicted arrhythmias in CUD while other substance abuse did not change the risk of arrhythmia in CUD. Arrhythmia burden is increasing among teenagers with CUD, and co-occurrence of arrhythmia and CUD worsens hospital outcomes.


Solvation and cavity occupation in biomolecules.
Solvation density locations are important for protein dynamics and structure. Knowledge of the preferred hydration sites at biomolecular interfaces and those in the interior of cavities can enhance understanding of structure and function. While advanced X-ray diffraction methods can provide accurate atomic structures for proteins, that technique is challenged when it comes to providing accurate hydration structures, especially for interfacial and cavity bound solvent molecules. Advances in integral equation theories which include more accurate methods for calculating the long-ranged Coulomb interaction contributions to the three-dimensional distribution functions make it possible to calculate angle dependent average solvent structure, accurately, around and inside irregular molecular conformations. The proximal radial distribution method provides another approximate method to determine average solvent structures for biomolecular systems based on a proximal or near neighbor solvent distribution that can be constructed from previously collected solvent distributions. These two approximate methods, along with all-atom molecular dynamics simulations are used to determine the solvent density inside the myoglobin heme cavity. Myoglobin is a good test system for these methods because the cavities are many and one is large, tens of Å(3), but is shown to have only four hydration sites. These sites are not near neighbors which implies that the large cavity must have more than one way in and out. Our results show that main solvation sites are well reproduced by all three methods. The techniques also produce a clearly identifiable solvent pathway into the interior of the protein. The agreement between molecular dynamics and less computationally demanding approximate methods is encouraging. This article is part of a Special Issue entitled Recent developments of molecular dynamics.


[Biosynthesis of 14C- and 35S-lincomycin using different sources for the label].
The sources of radioactive labels were chosen for biosynthesis of labeled lincomycin. The levels of the label incorporation into lincomycin were high with all the sources used when the lincomycin-producing organism was cultivated on the synthetic medium as compared to the complex medium. Incorporation of the label into lincomycin was most effective when I14C- or 214C-thyrosine was used as the precursors. These precursors were effective in both the complex and the synthetic media. In production of significant amounts of the labeled lincomycin I14C- or 214C-sodium propionate, I14C- or 142C-sodium acetate and 14C-protein hydrolysate may be used as the sources of the radioactive carbon. In production of 35S-lincomycin K235SO4 may be used as the source of the label. The optimal conditions for biosynthesis of 14C- and 35S-lincomycin were developed (concentration of some components of the medium, time of the label addition and others).


Effect of dried fruits of Solanum nigrum LINN against CCl4-induced hepatic damage in rats.
Ethanol extract of Solanum nigrum LINN was investigated for its hepatoprotective activity against CCl4-induced hepatic damage in rats. The ethanol extract showed remarkable hepatoprotective activity. The activity was evaluated using biochemical parameters such as serum aspartate amino transferase (AST), alanine amino transferase (ALT), alkaline phosphatase (ALP) and total bilirubin. The histopathological changes of liver sample in treated animals were compared with respect to control.


Analysis and consensus of currently available intrinsic protein disorder annotation sources in the MobiDB database.
Intrinsic protein disorder is becoming an increasingly important topic in protein science. During the last few years, intrinsically disordered proteins (IDPs) have been shown to play a role in many important biological processes, e.g. protein signalling and regulation. This has sparked a need to better understand and characterize different types of IDPs, their functions and roles. Our recently published database, MobiDB, provides a centralized resource for accessing and analysing intrinsic protein disorder annotations. Here, we present a thorough description and analysis of the data made available by MobiDB, providing descriptive statistics on the various available annotation sources. Version 1.2.1 of the database contains annotations for ca. 4,500,000 UniProt sequences, covering all eukaryotic proteomes. In addition, we describe a novel consensus annotation calculation and its related weighting scheme. The comparison between disorder information sources highlights how the MobiDB consensus captures the main features of intrinsic disorder and correlates well with manually curated datasets. Finally, we demonstrate the annotation of 13 eukaryotic model organisms through MobiDB's datasets, and of an example protein through the interactive user interface. MobiDB is a central resource for intrinsic disorder research, containing both experimental data and predictions. In the future it will be expanded to include additional information for all known proteins.


Neurons within the same network independently achieve conserved output by differentially balancing variable conductance magnitudes.
Biological and theoretical evidence suggest that individual neurons may achieve similar outputs by differentially balancing variable underlying ionic conductances. Despite the substantial amount of data consistent with this idea, a direct biological demonstration that cells with conserved output, particularly within the same network, achieve these outputs via different solutions has been difficult to achieve. Here we demonstrate definitively that neurons from native neural networks with highly similar output achieve this conserved output by differentially tuning underlying conductance magnitudes. Multiple motor neurons of the crab (Cancer borealis) cardiac ganglion have highly conserved output within a preparation, despite showing a 2-4-fold range of conductance magnitudes. By blocking subsets of these currents, we demonstrate that the remaining conductances become unbalanced, causing disparate output as a result. Therefore, as strategies to understand neuronal excitability become increasingly sophisticated, it is important that such variability in excitability of neurons, even among those within the same individual, is taken into account.


[Effect of vasopressin and enkephalin analogs on the chemoreactive properties of the sensorimotor cortex neurons in rats].
On immobile rats it has been shown that subcuteneous injection of two different neuropeptides causes different distribution of activating, inhibiting and areactive cells resulting from microionoforetically administrated acetylcholine and noradrenaline without altering the mean frequency of the background unit activity sensorimotor cortical neurons. At the same time the alteration in the pattern of neurons impulse activity has been revealed, which acquires the character of packet activity with a specific packet length of 2-3, 4-6, 7-10 impulses. Depending on the injected neuropeptide reliable but almost contrary effects of the packet activity alterations have been observed. These effects have been suggested to result from the alteration of the chemoreactive properties of the neuron membranes and the corresponding reorganisations of interneuron attitudes.


Electrocoagulation and the risk of cervical neoplasia.
The relationship between electrocoagulation diathermy and the risk of cervical neoplasia was evaluated in a case-control study of 145 women with cervical intraepithelial neoplasia compared with 145 age-matched outpatient control subjects, and 191 cases of invasive cervical cancer compared with 191 control subjects in the hospital for acute conditions unrelated to any of the identified or suspected risk factors for cervical cancer. History of electrocoagulation was associated with an apparently reduced risk of cervical intraepithelial neoplasia (relative risk = 0.50, with 95% confidence interval = 0.29-0.87), and of invasive cancer (relative risk = 0.42, 95% confidence interval = 0.22-0.82). However, this apparent protection could be largely explained in terms of a different frequency of previous Papanicolaou smears in patients and control subjects. When adjustment was made for that variable, the risk estimates of CIN and invasive cancer among women who had undergone electrocoagulation increased to 0.62 and 0.83 and became statistically nonsignificant. Further allowance for other identified potential confounding factors by means of multiple logistic regression raised these estimates to 0.73 and 0.94, respectively. Thus, these data provide evidence against the hypothesis that electrocoagulation may have an important and independent role in the prevention of cervical neoplasia.


Histochemical analysis of the development of estradiol-induced kidney tumors in male Syrian hamsters.
Chronic administration of the estrogen 17 beta-estradiol induces kidney tumors in male Syrian hamsters within 6 months of initial exposure. Although these tumors have previously been studied histologically and histochemically and have been postulated to be derived from proximal tubular and/or interstitial cells, there exists no unambiguous evidence for an epithelial or mesenchymal origin. To elucidate the histogenesis of these neoplasms, kidney sections of hamsters treated with estradiol for 4, 5, and 6 months and age-matched untreated controls were investigated histologically and histochemically. Proliferating foci were observed in kidneys exposed to estradiol for 5 and 6 months. They consisted of clusters of spindle-shaped cells forming solid blocks, cords, or branches located between tubules. These foci were judged to be precursors of larger tumors identified in the latter treatment group. The histological and histochemical profile of foci and tumors matched closely. These lesions were marked by very high activities of alkaline phosphatase, adenyl cyclase, and glucose 6-phosphate dehydrogenase. In contrast, glycogen content and activities of glucose 6-phosphatase, succinate dehydrogenase, and gamma-glutamyl transpeptidase were low or absent. Immunofluorescence of the intermediate filaments revealed that foci and tumors solely expressed vimentin and desmin but not cytokeratin. The morphology, enzyme histochemical pattern, and immunofluorescence strongly support a mesenchymal origin of the estradiol-induced hamster kidney tumors studied. The neoplasms were probably derived from vascular smooth muscle cells of a cell subtype particularly sensitive to hormonal stimulation and transformation.


Is suspicion of genital herpes infection associated with avoiding sex? A clinic-based study.
The present study tested the research hypothesis that sexually transmissible disease (STD) clinic patients suspecting genital herpes infection would be more likely than their 'non-suspecting' counterparts to abstain from sex to avoid transmission of a perceived STD. Recruitment (n = 351) occurred in a publicly-funded STD clinic located in a metropolitan area of the southern USA. Participants were tested for herpes simplex virus type 2 (HSV-2) using a rapid test manufactured by Biokit (Lexington, MA, USA) and they completed a self-administered questionnaire (using a 3-month recall period). Well over one-third (38.7%) of those indicating suspicion also indicated avoiding sex with steady partners because of concerns about STDs as compared with 28.0% among those not indicating suspicion (prevalence ratio = 1.38; 95% CI = 1.02-1.87, P = 0.036). The relationship between suspicion and avoiding sex with non-steady partners was not significant (P = 0.720). The relationship with steady partners only applied to people who were female (P = 0.013), single (P = 0.017), reported symptoms of genital herpes (P = 0.003), perceived that genital herpes would have a strong negative influence on their sex life (P = 0.0001), and who subsequently tested positive for HSV-2 (P = 0.012). Among STD clinic attendees, suspicion of genital herpes infection may translate into partner protective behaviour, but only for a minority of people and only with respect to sex with steady partners. Clinic-based and community-based education programs may benefit public health by teaching people (especially single women) how to effectively recognise symptoms of primary genital herpes infections. Reversing the often prevailing ethic of genital herpes as a 'community secret' will clearly be a challenge to these education programs.


Oxytocin-stimulated phosphoinositide hydrolysis and prostaglandin F2alpha secretion by luminal epithelial, glandular epithelial and stromal cells from pig endometrium. II. Responses of cyclic, pregnant and pseudopregnant pigs on days 12 and 16 post oestrus.
In pigs, the exact mechanism for the shift in endometrial PGF2alpha secretion from an endocrine to an exocrine mode during pregnancy recognition is not known. The objective of this study was to examine whether this shift involved a change in the responsiveness of luminal epithelial, glandular epithelial and stromal cells to 0 or 100 nM oxytocin. Luminal epithelial cells, glandular epithelial cells and stromal cells were isolated from cyclic, pregnant or oestrogen-induced pseudopregnant gilts on Day 12 (Experiment 1) or Day 16 (Experiment 2) post oestrus (oestrus = Day 0). For cells obtained on Day 12, oxytocin stimulated PGF2alpha secretion by stromal cells (P<0.01) similarly for each reproductive status, whereas oxytocin stimulated PGF2alpha secretion from luminal and glandular epithelial cells (P<0.05) from pregnant and pseudopregnant gilts but not from cyclic gilts. For both concentrations of oxytocin, mean PGF2alpha secretion was less (P<0.05) from stromal cells of pregnant than cyclic gilts. For cells obtained on Day 16, oxytocin stimulated PGF2alpha release from stromal cells of cyclic gilts but not from stromal cells of pregnant gilts. Mean PGF2alpha secretion also was less (P<0.05) from stromal cells of pregnant gilts than cyclic gilts. Oxytocin tended to stimulate PGF2alpha release (P<0.07) from glandular epithelial cells of cyclic but not pregnant or pseudopregnant gilts. Luminal epithelial cells from all reproductive statuses were similarly unresponsive to oxytocin. In conclusion, the increased PGF2alpha secretory response to oxytocin of luminal and glandular epithelial cells from pregnant gilts on Day 12, combined with the decreased response of stromal cells from pregnant gilts on Days 12 and 16, may contribute, in part, to the shift in endometrial PGF2alpha secretion from an endocrine to an exocrine direction during early pregnancy in pigs.


Moderate exercise triggers both priming and activation of neutrophil subpopulations.
We investigated how moderate exercise affects neutrophil microbicidal activity and whether exercise-induced responses are associated with changes in growth hormone (GH) secretion. Biological fluctuations were controlled for and GH secretion was manipulated by glucose ingestion. In eight men, 1 h of moderate exercise increased intracellular H2O2 generation in response to phorbol 12-myristate 13-acetate stimulation by threefold (P = 0.025) and complement receptor expression by 20% (P = 0.045). These responses were accompanied by a twofold increase in the plasma concentration of elastase, a marker of neutrophil activation in vivo. The plasma concentration of GH increased 10-fold after exercise, but this was reduced to 3-fold by glucose ingestion (P < 0.001), which also blunted elastase release (P < 0.001). Although the magnitude of H2O2 generation increased in proportion to the increase in plasma GH concentration, it declined progressively once this exceeded 20 ng/ml. The net response of neutrophils to exercise may represent a balance between the individual responses of subpopulations that are unaffected, primed, or fully activated by circulating mediators that respond to exercise and to dietary glucose intake.


Combination of electrospray ionization, atmospheric pressure photoionization and laser desorption ionization Fourier transform ion cyclotronic resonance mass spectrometry for the investigation of complex mixtures - Application to the petroleomic analysis of bio-oils.
The comprehensive description of complex mixtures such as bio-oils is required to understand and improve the different processes involved during biological, environmental or industrial operation. In this context, we have to consider how different ionization sources can improve a non-targeted approach. Thus, the Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) has been coupled to electrospray ionization (ESI), laser desorption ionization (LDI) and atmospheric pressure photoionization (APPI) to characterize an oak pyrolysis bio-oil. Close to 90% of the all 4500 compound formulae has been attributed to CxHyOz with similar oxygen class compound distribution. Nevertheless, their relative abundance in respect with their double bound equivalent (DBE) value has evidenced significant differences depending on the ion source used. ESI has allowed compounds with low DBE but more oxygen atoms to be ionized. APPI has demonstrated the efficient ionization of less polar compounds (high DBE values and less oxygen atoms). The LDI behavior of bio-oils has been considered intermediate in terms of DBE and oxygen amounts but it has also been demonstrated that a significant part of the features are specifically detected by this ionization method. Thus, the complementarity of three different ionization sources has been successfully demonstrated for the exhaustive characterization by petroleomic approach of a complex mixture.


Piloting an integrated SARS-CoV-2 testing and data system for outbreak containment among college students: A prospective cohort study.
Colleges and universities across the country are struggling to develop strategies for effective control of COVID-19 transmission as students return to campus. We conducted a prospective cohort study with students living on or near the UC Berkeley campus from June 1st through August 18th, 2020 with the goal of providing guidance for campus reopening in the safest possible manner. In this cohort, we piloted an alternative testing model to provide access to low-barrier, high-touch testing and augment student-driven testing with data-driven adaptive surveillance that targets higher-risk students and triggers testing notifications based on reported symptoms, exposures, or other relevant information. A total of 2,180 students enrolled in the study, 51% of them undergraduates. Overall, 6,247 PCR tests were administered to 2,178 students over the two-month period. Overall test positivity rate was 0.9%; 2.6% of students tested positive. Uptake and acceptability of regular symptom and exposure surveys was high; 98% of students completed at least one survey, and average completion rate was 67% (Median: 74%, IQR: 39%) for daily reporting of symptoms and 68% (Median: 75%, IQR: 40%) for weekly reporting of exposures. Of symptom-triggered tests, 5% were PCR-positive; of exposure-triggered tests, 10% were PCR-positive. The integrated study database augmented traditional contact tracing during an outbreak; 17 potentially exposed students were contacted the following day and sent testing notifications. At study end, 81% of students selected their desire "to contribute to UC Berkeley's response to COVID-19" as a reason for their participation in the Safe Campus study. Our results illustrate the synergy created by bringing together a student-friendly, harm reduction approach to COVID-19 testing with an integrated data system and analytics. We recommend the use of a confidential, consequence-free, incentive-based daily symptom and exposure reporting system, coupled with low-barrier, easy access, no stigma testing. Testing should be implemented alongside a system that integrates multiple data sources to effectively trigger testing notifications to those at higher risk of infection and encourages students to come in for low-barrier testing when needed.


Effects of Eight Interval Training Sessions in Hypoxia on Anaerobic, Aerobic, and High Intensity Work Capacity in Endurance Cyclists.
Aim: This study aimed to determine if eight sessions of supramaximal but steady-state, set duration interval training in hypoxia enhanced measured anaerobic capacity and work performed during high intensity exercise. High Alt Med Biol. 21:370-377, 2020. Materials and Methods: Eighteen cyclists (V̇O2peak: 57 ± 7 ml·kg-1·min-1) were pair-matched for anaerobic capacity determined by maximal accumulated oxygen deficit (MAOD) and allocated to a 4-week interval training in hypoxia (IHT; FiO2 = 14.7% ± 0.5%, n = 9) or interval training in normoxia (NORM; FiO2 = 20.6% ± 0.3%, n = 9). Cyclists completed twice weekly interval training (8 × 1 minutes: ∼120% V̇O2peak, 5 minutes recovery: ∼50% V̇O2peak) in addition to their habitual training. Before and after the intervention, a constant work rate supramaximal time to fatigue and a graded exercise test were used to determine changes in anaerobic capacity/supramaximal work performed and aerobic capacity/peak aerobic power output, respectively. Results: No interaction or main effects were observed. Using indirect calorimetry, anaerobic capacity was not significantly different in either group pre- to postintervention using MAOD (IHT: 4% ± 15%; NORM: -5% ± 12%) or gross efficiency methods (IHT: 7% ± 14%; NORM: -2% ± 9%), and VO2peak was unchanged (IHT: 1% ± 6%; NORM: 1% ± 4%). However, within-group analysis shows that supramaximal work performed improved with IHT (14% ± 13%; p = 0.02; d = 0.42) but not NORM (1% ± 22%), and peak aerobic power output increased with IHT (5% ± 7%; p = 0.04; d = 0.32) but not NORM (2% ± 4%). Conclusion: Steady-state, set duration supramaximal interval training in hypoxia appears to provide a small beneficial effect on work capacity during supramaximal and high intensity exercise.


B-DNA structure and stability: the role of hydrogen bonding, π-π stacking interactions, twist-angle, and solvation.
We have computationally investigated the structure and stability of B-DNA. To this end, we have analyzed the bonding in a series of 47 stacks consisting of two base pairs, in which the base pairs cover the full range of natural Watson-Crick pairs, mismatched pairs, and artificial DNA base pairs. Our analyses provide detailed insight into the role and relative importance of the various types of interactions, such as, hydrogen bonding, π-π stacking interactions, and solvation/desolvation. Furthermore, we have analyzed the functionality of the twist-angle on the stability of the structure. Interestingly, we can show that all stacked base pairs benefit from a stabilization by 6 to 12 kcal mol(-1) if stacked base pairs are twisted from 0° to 36°, that is, if they are mutually rotated from a congruent superposition to the mutually twisted stacking configuration that occurs in B-DNA. This holds especially for stacked AT pairs but also for other stacked base pairs, including GC. The electronic mechanism behind this preference for a twisted arrangement depends on the base pairs involved. We also show that so-called "diagonal interactions" (or cross terms) in the stacked base pairs are crucial for understanding the stability of B-DNA, in particular, in GC-rich sequences.


Distal forelimb representations in primary motor cortex are redistributed after forelimb restriction: a longitudinal study in adult squirrel monkeys.
Primary motor cortex (M1) movement representations reflect acquired motor skills. Representations of muscles and joints used in a skilled task expand. However, it is unknown whether motor restriction in healthy individuals results in complementary reductions in M1 representations. With the use of intracortical microstimulation techniques in squirrel monkeys, detailed maps of movement representations in M1 were derived before and up to 35 wk after restriction of the preferred distal forelimb (DFL) by use of a soft cast. Although total DFL area and movement threshold remained constant, casting resulted in a redistribution of digit and wrist/forearm representations. Digit representations progressively decreased, whereas wrist/forearm representations progressively increased in areal extent. In three of four monkeys, hand preference returned to normal by the end of the postcast recovery period, and postrecovery maps demonstrated reversal of restriction-induced changes. However, in one monkey, a chronic motor impairment occurred in the casted limb. Rehabilitation via a forced-use paradigm resulted in recovery in use and skill of the impaired limb, as well as restoration of normal motor maps. These results demonstrate that plasticity in motor representations can be induced by training or restricting movements of the limb. Physiological changes induced by restriction appear to be reversible, even in the case of adverse motor outcomes. The respective contributions of both disuse and lost motor skills are discussed. These results have relevance for clinical conditions requiring forelimb casting as well as interpreting the differential effects of injury and disuse that are necessarily intertwined after cortical injury, as occurs in stroke.


Hybrid Doping of Few-Layer Graphene via a Combination of Intercalation and Surface Doping.
Surface molecular doping of graphene has been shown to modify its work function and increase its conductivity. However, the associated shifts in work function and increases in carrier concentration are highly coupled and limited by the surface coverage of dopant molecules on graphene. Here we show that few-layer graphene (FLG) can be doped using a hybrid approach, effectively combining surface doping by larger (metal-)organic molecules and intercalation of smaller molecules, such as Br2 and FeCl3, into the bulk. Intercalation tunes the carrier concentration more effectively, whereas surface doping of intercalated FLG can be used to tune its work function without reducing the carrier mobility. This multimodal doping approach yields a very high carrier density and tunable increase in the work function for FLG, demonstrating a new versatile platform for fabricating graphene-based contacts for electronic, optoelectronic, and photovoltaic applications.