Hydration of porphyrin and Mg-porphyrin: ab initio quantum mechanical charge field molecular dynamics simulations.
Ab initio QMCF-MD simulations were performed for porphyrin (POR) and magnesium-porphyrin (Mg-POR) immersed in water to study their structural and dynamical properties. The observed hydration behaviour of these solutes representing biomimetic models is in fair agreement with structural and dynamical features of their biological analogues, protoporphyrin IX (PPIX) and chlorophyll (CHl). Structural data obtained from the radial, angular and spatial distribution functions as well as the angular-radial distributions have a consensus on possessing a contrasting hydration behaviour of POR and Mg-POR. Flexibility of the ring in both solutes described by the improper torsional distribution and root mean square fluctuation showed an influence on H-bond interactions between the nitrogen atoms and water molecules that are also reflected in the respective dynamics. An axial water molecule coordinated to the Mg(ii) ion indicates the penta-coordinated Mg-POR to be stable along the simulation. It was also shown that complexation of the Mg(ii) ion to the porphyrin influences the hydration patterns significantly compared to the porphyrin itself, which is further supported by the vibrational power spectra evaluated for both solutes. Free energy of binding and solvent accessible surface area calculations also confirmed that these two solutes have distinct hydration behaviour. Detailed knowledge of the individual hydration patterns is expected to be of particular benefit.


Macrophage migration inhibitory factor induces autophagy via reactive oxygen species generation.
Autophagy is an evolutionarily conserved catabolic process that maintains cellular homeostasis under stress conditions such as starvation and pathogen infection. Macrophage migration inhibitory factor (MIF) is a multifunctional cytokine that plays important roles in inflammation and tumorigenesis. Cytokines such as IL-1β and TNF-α that are induced by MIF have been shown to be involved in the induction of autophagy. However, the actual role of MIF in autophagy remains unclear. Here, we have demonstrated that incubation of human hepatoma cell line HuH-7 cells with recombinant MIF (rMIF) induced reactive oxygen species (ROS) production and autophagy formation, including LC3-II expression, LC3 punctae formation, autophagic flux, and mitochondria membrane potential loss. The autophagy induced by rMIF was inhibited in the presence of MIF inhibitor, ISO-1 as well as ROS scavenger N-acetyl-L-cysteine (NAC). In addition, serum starvation-induced MIF release and autophagy of HuH-7 cells were partly blocked in the presence of NAC. Moreover, diminished MIF expression by shRNA transfection or inhibition of MIF by ISO-1 decreased serum starvation-induced autophagy of HuH-7 cells. Taken together, these data suggest that cell autophagy was induced by MIF under stress conditions such as inflammation and starvation through ROS generation.


Transferrin binding of Al3+ and Fe3+.
An understanding of Al3+-induced diseases requires identification of the blood carrier of Al3+ to the tissues where Al3+ exerts a toxic action. Quantitative studies demonstrate that the protein transferrin (iron-free) is the strongest Al3+ binder in blood plasma. Under plasma conditions of pH 7.4 and [HCO3-]27 mmol/L, the successive stability constant values for Al3+ binding to transferrin are log K1 = 12.9 and log K2 = 12.3. When the concentration of total Al3+ in plasma is 1 mumol/L, the free Al3+ concentration permitted by transferrin is 10(-14.6) mol/L, less than that allowed by insoluble Al(OH)3, by Al(OH)2H2PO4, or by complexing with citrate. Thus transferrin is the ultimate carrier of Al3+ in the blood. We also used intensity changes produced by metal ion binding to determine the stability constants for Fe3+ binding to transferrin: log K1 = 22.7 and log K2 = 22.1. These constants agree closely with a revision of the reported values obtained by equilibrium dialysis. By comparison with Fe3+ binding, the Al3+ stability constants are weaker than expected; this suggests that the significantly smaller Al3+ ions cannot coordinate to all the transferrin donor atoms available to Fe3+.


An extrafollicular pathway for the generation of effector CD8(+) T cells driven by the proinflammatory cytokine, IL-12.
The proinflammatory cytokine IL-12 drives the generation of terminally differentiated KLRG1(+) effector CD8(+) T cells. Using a Toxoplasma vaccination model, we delineate the sequence of events that naïve CD8(+) T cells undergo to become terminal effectors and the differentiation steps controlled by IL-12. We demonstrate that direct IL-12 signaling on CD8(+) T cells is essential for the induction of KLRG1 and IFN-γ, but the subsequent downregulation of CXCR3 is controlled by IL-12 indirectly through the actions of IFN-γ and IFN-γ-inducible chemokines. Differentiation of nascent effectors occurs in an extrafollicular splenic compartment and is driven by late IL-12 production by DCs distinct from the classical CD8α(+) DC. Unexpectedly, we also found extensive proliferation of both KLRG1(-) and KLRG1(+) CD8(+) T cells in the marginal zone and red pulp, which ceases prior to the final KLRG1(Hi) CXCR3(Lo) stage. Our findings highlight the notion of an extrafollicular pathway for effector T cell generation.


Interaction of three second-generation anthracyclines with polynucleotides, RNA, DNA, and nucleosomes.
The interaction of three second-generation anthracycline derivatives with polynucleotides, supercoiled DNA, and calf thymus nucleosomes has been studied by terbium fluorescence measurements and agarose gel electrophoresis. It was shown that, as expected, N-trifluoroacetyladriamycin-14-valerate had little detectable effect on the fluorescence of this guanine-specific probe except for polydisperse calf thymus linear DNA fragments. The soluble analogue, N-trifluoroacetyladriamycin-14-O-hemiadipate, did show marked effects on terbium fluorescence with all nucleic acids and nucleosomes, but the effects were generally not as striking as were those observed with the epimer, 4'-epi-Adriamycin, which tended to produce a similar effect to its parent drug, Adriamycin, showing that a marked change in the hexose ring did not appreciably affect the interaction of the drug with DNA. Changes in the electrophoretic mobility of supercoiled pBR322 DNA were observed only at very high drug concentrations, much higher than those required with Adriamycin or actinomycin D. The effect was a smearing of the form I DNA and the production of some circular relaxed form II DNA. The drugs produced the effect in the following order: Adriamycin greater than N-trifluoroacetyladriamycin-14-O-hemiadipate greater than or equal to epi-Adriamycin. N-Trifluoroacetyladriamycin-14-valerate had little effect, even at very high drug concentrations (1:2, drug:DNA ratios).


T lymphocytes infiltrating sites of tumor rejection and progression display identical V beta usage but different cytotoxic activities.
Most tumors grow progressively and overwhelm the host. The rare but documented cases of spontaneous regression of primary tumors are indicative of the potential of tumor-bearing hosts to develop a significant antitumor response. Because most tumors grow progressively in the host, it is not surprising that the majority of studies have focused on T lymphocytes that infiltrate these tumors. Although these studies have generated significant and useful information during the period of tumor growth, they can only speculate on the mechanisms that are involved in tumor rejection. We have used a well developed sponge model of concomitant tumor immunity that allows us to compare the immunologic events that occur during tumor progression vs rejection. In this model, an animal harboring a primary EMT6 mammary tumor is challenged with a secondary tumor implant through a pre-implanted gelatin sponge. During the manifestation of concomitant tumor immunity, the secondary tumor is rejected and the effector cells mediating the response are retained within the sponge matrix. Using this model we analyzed the TCR usage, cytotoxic activity of lymphocytes, and cytokine production at both tumor sites. The data revealed that tumor-rejecting lymphocytes isolated from the site of secondary tumor implant were cytotoxic toward EMT6 cells, whereas tumor-infiltrating lymphocytes isolated from the progressing primary tumor were not. Interestingly, the TCR-V beta repertoire of the tumor-infiltrating lymphocytes and tumor-rejecting lymphocytes were identical with V beta 1 and V beta 8 being predominant at both sites. Furthermore, the rejection site showed higher gene expression of IFN-gamma, TNF-alpha, and IL-10 whereas TGF-beta expression was slightly higher in the progressing tumors. These findings suggest that the disparate effector functions observed during tumor progression vs rejection are not caused by different T cell phenotypes but may be due instead to influences exerted by cytokines produced at the tumor sites.


Clinical outcomes and survival analysis for petroclival meningioma patients receiving surgical resection: an analysis of 176 cases.
Purpose: The relationship of clinical results and survival analysis of operative patients with petroclival meningioma (PCM) was studied. Patients and methods: Data from a total of 176 PCM patients receiving surgical resection were retrospectively collected. Follow-up was conducted through outpatient review by reexamination telephone calls and letters. Clinical outcomes, survival, and Karnofsky Performance Scale (KPS) data were analyzed. Results: Seventy-two percent of patients (127/176) received only surgery, 8.5% (15/176) received surgery and adjuvant radiation therapy (RT) (surgery + RT), and 19.3% (34/176) received surgery and adjuvant gamma knife surgery (GKS) (surgery + GKS). Gross total resection (GTR) was performed in 34.7% of patients (61/176), subtotal resection (STR) in 58.0% (102/176), and partial resection (PR) in 7.4% (13/176). Recent follow-up KPS was higher than preoperative (80 vs 70, P<0.05) and postoperative KPS (80 vs 70, P<0.05). Cumulative survival was the same for 3, 5, 7, and 9 years, ie, 95%, and mean survival time (MST) was (110.83±2.55) months (95% CI: 105.83-115.83). Recurrence/progression (R/P)-free survival was 88.9%, 86.9%, 71.1% and 71.1%, respectively for 3, 5, 7, and 9 years, and MST was (100.58±3.82) months (95% CI: 93.11-108.06). R/P (HR: 5.486, 95% CI: 1.655-18.180), surgery + RT (HR: 0.125, 95% CI: 0.016-0.990) and WHO grade III (HR: 2.766, 95% CI: 1.146-6.676) were independently associated with cumulative survival. Lack of adhesion to and encasement of neurovascular structures was independently associated with R/P-free survival (HR: 2.002, 95% CI: 1.023-3.919). Conclusion: Surgical treatment was safe and effective for PCM. R/P, surgery + RT, and WHO grade III were independently associated with cumulative survival. Lack of adhesion to and encasement of neurovascular structures was independently associated with R/P-free survival. These factors should be paid attention to in surgical treatment of PCM.


Evaluation of radiographic predictors of left heart enlargement in dogs with known or suspected cardiovascular disease.
Radiographic assessment of heart size is important for clinical management of dogs with cardiovascular disease (CVDz). We sought to compare the ability of vertebral heart size (VHS), vertebral left atrial size (VLAS), and radiologists' assessment of left atrial size (RadLAE) to predict echocardiographic left atrial size (EchoLAE), an important marker of left heart disease severity. We also compared the ability of VHS and VLAS to predict echocardiographic criteria for ACVIM stage B2 (EchoB2) in dogs with myxomatous mitral valve disease (MMVD). This prospective observational study enrolled 183 dogs with known or suspected CVDz that had an echocardiographic examination and thoracic radiographs obtained within 24 h. Compared to increased VHS, VLAS >2.3 was a more accurate predictor of EchoLAE (P = .002). VLAS >2.3 and RadLAE (both P <.0001) were independently associated with EchoLAE but VHS was not (P = .45). Optimal cutoffs for VLAS and VHS to predict EchoLAE were >2.3 vertebrae (sensitivity [Sn] = 90.3%, specificity [Sp] = 73.6%) and >11.1 vertebrae (Sn = 75.8%, Sp = 76.0%), respectively. Diagnostic accuracy of VLAS (AUC 0.84, 95% CI 0.73-0.92) and VHS (AUC 0.78, 95% CI 0.66-0.88) to predict EchoB2 in dogs with subclinical MMVD (n = 64) were not significantly different (P = .17). Results demonstrate that VLAS and RadLAE were superior indicators of EchoLAE compared to VHS in dogs with known or suspected CVDz. Both VLAS and VHS are useful predictors of EchoB2 in dogs with subclinical MMVD. When echocardiography is unavailable, VLAS represents a useful radiographic measurement to aid clinical management of dogs with known or suspected CVDz.


Ketamine normalizes brain activity during emotionally valenced attentional processing in depression.
An urgent need exists for faster-acting pharmacological treatments in major depressive disorder (MDD). The glutamatergic modulator ketamine has been shown to have rapid antidepressant effects, but much remains unknown about its mechanism of action. Functional MRI (fMRI) can be used to investigate how ketamine impacts brain activity during cognitive and emotional processing. This double-blind, placebo-controlled, crossover study of 33 unmedicated participants with MDD and 26 healthy controls (HCs) examined how ketamine affected fMRI activation during an attentional bias dot probe task with emotional face stimuli across multiple time points. A whole brain analysis was conducted to find regions with differential activation associated with group, drug session, or dot probe task-specific factors (emotional valence and congruency of stimuli). A drug session by group interaction was observed in several brain regions, such that ketamine had opposite effects on brain activation in MDD versus HC participants. Additionally, there was a similar finding related to emotional valence (a drug session by group by emotion interaction) in a large cluster in the anterior cingulate and medial frontal cortex. The findings show a pattern of brain activity in MDD participants following ketamine infusion that is similar to activity observed in HCs after placebo. This suggests that ketamine may act as an antidepressant by normalizing brain function during emotionally valenced attentional processing. NCT#00088699: https://www.clinicaltrials.gov/ct2/show/NCT00088699.


Safety, efficacy and physiological actions of a lysine-free, arginine-rich formula to treat glutaryl-CoA dehydrogenase deficiency: focus on cerebral amino acid influx.
Striatal degeneration from glutaryl-CoA dehydrogenase deficiency (glutaric aciduria type 1, GA1) is associated with cerebral formation and entrapment of glutaryl-CoA and its derivatives that depend on cerebral lysine influx. In 2006 we designed a lysine-free study formula enriched with arginine to selectively block lysine transport across cerebral endothelia and thereby limit glutaryl-CoA production by brain. Between 2006 and present, we treated twelve consecutive children with study formula (LYSx group) while holding all other treatment practices constant. Clinical and biochemical outcomes were compared to 25 GA1 patients (PROx group) treated between 1995 and 2005 with natural protein restriction (dietary lysine/arginine ratio of 1.7±0.3 mg:mg). We used published kinetic parameters of the y+and LAT1 blood-brain barrier transporters to model the influx of amino acids into the brain. Arginine fortification to achieve a mean dietary lysine/arginine ratio of 0.7±0.2 mg:mg was neuroprotective. All 12 LYSx patients are physically and neurologically healthy after 28 aggregate patient-years of follow up (current ages 28±21 months) and there were no adverse events related to formula use. This represents a 36% reduction of neurological risk (95% confidence interval 14-52%, p=0.018) that we can directly attribute to altered amino acid intake. During the first year of life, 20% lower lysine intake and two-fold higher arginine intake by LYSx patients were associated with 50% lower plasma lysine, 3-fold lower plasma lysine/arginine concentration ratio, 42% lower mean calculated cerebral lysine influx, 54% higher calculated cerebral arginine influx, 15-26% higher calculated cerebral influx of several anaplerotic precursors (isoleucine, threonine, methionine, and leucine), 50% less 3-hydroxyglutarate excretion, and a 3-fold lower hospitalization rate (0.8 versus 2.3 hospitalizations per patient per year). The relationship between arginine fortification and plasma lysine indicates that transport competition exists at both cerebrovascular and gastrointestinal barriers, suggesting their co-administration is key to efficacy. Monitoring the ratio between lysine and arginine in diet and plasma may prove a useful strategy for treating children with GA1.


A Literature Review of the Pharmacokinetics and Pharmacodynamics of Dobutamine in Neonates.
Since its discovery in 1975 dobutamine has been used off-label for treating hemodynamic insufficiency in newborns and children. We present a structured literature review of pharmacokinetic and pharmacodynamic data for dobutamine in the pediatric population. Structured searches were conducted to identify relevant articles according to pre-defined inclusion criteria. Where possible, results for the pharmacodynamic and pharmacokinetic effect of dobutamine were reported as pooled data. Forty-six papers met the inclusion criteria. With regard to pharmacodynamic data a number of studies reported significant increases in a number of clinical parameters such as heart rate, blood pressure, cardiac output across a wide range of pediatric populations. With regard to pharmacokinetic data studies reported that the infusion rate was positively correlated to plasma dobutamine concentration. There was great variability with regard to dobutamine clearance between individuals and as to whether it followed first- or zero-order elimination kinetics. While the pharmacodynamic effects of dobutamine appear to reflect the pharmacological profile of the drug, the pharmacokinetic data are difficult to interpret due to inhomogeneity between study populations ages, comorbidities, dobutamine dosages and methodologies. High-quality prospective pharmacokinetic and pharmacodynamic data especially in newborns are urgently required prior to a large randomized study.


Ability of two natural products, nootkatone and carvacrol, to suppress Ixodes scapularis and Amblyomma americanum (Acari: Ixodidae) in a Lyme disease endemic area of New Jersey.
We evaluated the ability of the natural, plant-derived acaricides nootkatone and carvacrol to suppress Ixodes scapularis Say and Amblyomma americanum (L.) (Acari: Ixodidae). Aqueous formulations of 1 and 5% nootkatone applied by backpack sprayer to the forest litter layer completely suppressed I. scapularis nymphs through 2 d. Thereafter, the level of reduction gradually declined to < or =50% at 28 d postapplication. Against A. americanum nymphs, 1% nootkatone was less effective, but at a 5% concentration, the level of control was similar or greater to that observed with I. scapularis through 21 d postapplication. Initial applications of 0.05% carvacrol were ineffective, but a 5% carvacrol formulation completely suppressed nymphs of both species through 2 d and resulted in significant reduction in I. scapularis and A. americanum nymphs through 28 and 14 d postapplication, respectively. Backpack sprayer applications of 5% nootkatone to the shrub and litter layers resulted in 100% control of I. scapularis adults through 6 d, but the level of reduction declined to 71.5% at 28 d postapplication. By contrast, high-pressure applications of 2% nootkatone to the litter layer resulted in 96.2-100% suppression of both I. scapularis and A. americanum nymphs through 42 d, whereas much lower control was obtained from the same formulation applied by backpack sprayer. Backpack sprayer application of a 3.1% nootkatone nanoemulsion resulted in 97.5-98.9 and 99.3-100% reduction in I. scapularis and A. americanum nymphs, respectively, at 1 d postapplication. Between 7 d and 35 d postapplication, the level of control varied between 57.1% and 92.5% for I. scapularis and between 78.5 and 97.1% for A. americanum nymphs. The ability of natural products to quickly suppress and maintain significant control of populations of these medically important ticks at relatively low concentrations may represent a future alternative to the use of conventional synthetic acaricides.


[Perinatal result of an institutional assisted reproduction program].
Assisted reproductive technology manipulates masculine gametes, embryos and implantation. It also aids the known or unknown factor of sterility without having the base problem correction as a target. In vitro fertilization and embryo transfer are among these techniques. To describe the overall outcome and the final perinatal offspring after in vitro fertilization cycle in an institutional third level hospital. IVF cycles were retrospectively analyzed from October 1999 to May 2004. Several variables were described like: age, fertilization rate, implantation and pregnancy rate, fetal status, time of gestation during labor, miscarriage follicle-stimulating hormone rate and take-home baby rate. Patients underwent hypophyseal supression with GnRH analog, using a long luteal phase protocol and stimulated with recombinant FSH. Overall data is expressed as average +/- standard deviation and percentage. 365 cycles were analyzed in 314 patients, average age was of 34 +/- 3.7 years, tubal factor was diagnosed in 63.3%, fertilization rate was of 60.4%, implantation rate of 37.1%, per transfer pregnancy rate of 25.1%, per transfer live born rate of 21.7%, multiple pregnancy rate of 29.3%, miscarriage rate of 28% and ectopic pregnancy rate of 4.8%. In 87.8% of the cases caesarean operation was made; multiple pregnancy offspring weighted more than 1250 g in 70% of them; 70.5% was born after 32 weeks of pregnancy; 90% was born live and with good prognosis. Glucose metabolism alterations were the most frequent maternal complication. Although the results obtained were similar to those of assisted reproduction centers, it suggests improving multiple pregnancy rate and abortion rate.


The forced swim test as a model of depressive-like behavior.
The goal of the present protocol is to describe the forced swim test (FST), which is one of the most commonly used assays for the study of depressive-like behavior in rodents. The FST is based on the assumption that when placing an animal in a container filled with water, it will first make efforts to escape but eventually will exhibit immobility that may be considered to reflect a measure of behavioral despair. This test has been extensively used because it involves the exposure of the animals to stress, which was shown to have a role in the tendency for major depression. Additionally, the FST has been shown to share some of the factors that are influenced or altered by depression in humans, including changes in food consumption, sleep abnormalities and drug-withdrawal-induced anhedonia. The main advantages of this procedure are that it is relatively easy to perform and that its results are easily and quickly analyzed. Moreover, its sensitivity to a broad range of antidepressant drugs that makes it a suitable screening test is one of the most important features leading to its high predictive validity. Despite its appeal, this model has a number of disadvantages. First, the issue of chronic augmentation is problematic in this test because in real life patients need to be treated for at least several weeks before they experience any relief from their symptoms. Last, due to the aversiveness of the FST, it is important to take into account possible influences it might have on brain structure/function if brain analyses are to be carried out following this procedure.


Antibodies against cyclic citrullinated peptides of IgG, IgA and IgM isotype and rheumatoid factor of IgM and IgA isotype are increased in unaffected members of multicase rheumatoid arthritis families from northern Sweden.
Rheumatoid factors (RFs) and antibodies against cyclic citrullinated peptides (CCPs) of IgG, IgA and IgM isotype have been shown to precede disease onset by years. To evaluate serological risk markers in first-degree relatives from multicase families in relation to genetic and environmental risk factors. 51 multicase families consisting of 163 individuals with rheumatoid arthritis (RA) (mean±SD age, 60±14 years; disease duration 21 years; 71.8% female) and with 157 first-degree relatives unaffected by RA (54±17 years; 59.9% female) were recruited. Isotypes of antibodies against CCPs (IgG, IgA and IgM) and RFs (IgM and IgA) were determined using automated enzyme immunoassays. Cut-off levels were established using receiver operating characteristic curves based on values for 100 unrelated healthy controls. The concentrations and frequencies of all anti-CCP and RF isotypes were significantly increased in first-degree relatives and patients with RA compared with unrelated healthy controls. The relative distribution of IgA and IgM isotypes was higher than IgG in the relatives, whereas the IgG isotype dominated in patients with RA. The patients carried human leucocyte antigen-shared epitope (HLA-SE) significantly more often than the relatives (71.4% vs 53.9%, p=0.01), while the frequency of the PTPN22 T variant was similar. HLA-SE, combined with smoking, was significantly related to all combinations of anti-CCP and RF isotypes in patients with RA. No such relationships were found for the first-degree relatives. All anti-CCP and RF isotypes analysed occurred more commonly in unaffected first-degree relatives from multicase families than in controls, but with different isotype distribution from patients with RA.


Factors driving bumble bee (Hymenoptera: Apidae: Bombus) and butterfly (Lepidoptera: Rhopalocera) use of sheared shrubland and young forest communities of the western Great Lakes.
In the northern Great Lakes region, the creation and maintenance of early-successional woody communities as wildlife habitat have increasingly become a conservation priority. The extent to which insect pollinators use these systems remains largely anecdotal. In summer (June-August) of 2021, we surveyed 49 early-successional sites in the western Great Lakes region treated with either shrub-shearing or silviculture (young forest) for bumble bees, butterflies, and habitat components (i.e., structural vegetation and floral resources). Hierarchical distance models predicted pollinator densities (λ^) to be, on average, λ^ = 84 bumble bees/ha and λ^ = 102 butterflies/ha. Although sheared shrubland and young forest communities supported comparable densities of bumble bees and butterflies, density was not equal across all sites. At the microhabitat scale, butterfly density and morphospecies richness were negatively associated with tall shrub cover and butterfly morphospecies richness (but not density) was driven by floral richness. Similarly, bumble bee density was positively associated with metrics of floral resources, underscoring the importance of blooming plants within these woody systems. Landscape covariates explained variation in butterfly density/richness but not bumble bee density. Ultimately, our results demonstrate that blooming plant abundance is an important driver of bumble bee and butterfly densities within these managed early-successional communities. Because early-successional woody communities are dynamic and their herbaceous openings are ephemeral, routine management would ensure that a variety of successional conditions exist on the landscape to meet the needs of bumble bees, butterflies, and potentially other insect pollinators.


[Protein metabolism in vegans].
To elucidate the mechanisms of adaptation to a low-energy and low-protein vegan diet, we carried out dietary surveys and nitrogen balance studies five times during one year on two women and a man who ate raw brown rice, raw green vegetables, three kinds of raw roots, fruit and salt daily. Individual subjects modified this vegan diet slightly. The mean daily energy intake of the subjects was 18, 14, and 32 kcal/kg, of body weight. The loss of body weight was about 10% of the initial level. The daily nitrogen balance was -32, -33, and -11 mg N/kg of body weight. In spite of the negative nitrogen balance, the results of routine clinical tests, initially normal, did not change with the vegan diet. Ten months after the start of the vegan diet, the subjects were given 15N urea orally. The incorporation of 15N into serum proteins suggested that these subjects could utilize urea nitrogen for body protein synthesis. The level of 15N in serum proteins was close to the level in other normal adult men on a low-protein diet with adequate energy for 2 weeks.


Computer-aided drug design course for pharmacy major students in Shenyang Pharmaceutical University following the COVID-19 pandemic: Challenges and opportunities.
The computer-aided drug design (CADD) course that spans biochemistry, computational chemistry, medicinal chemistry, and other cutting-edge sciences is considered an important course by pharmaceutical universities in China. The course teaches students how drugs bind to protein targets and exert their biological activities using computer tools, and covers the basic principles of drug development and optimization. Due to the lockdown and social distancing measures adopted during the coronavirus disease 2019 (COVID-19) pandemic, the CADD course in Shenyang Pharmaceutical University was briefly suspended. Thereafter, it was taught in the online mode by adopting a novel blended teaching method. Through a questionnaire survey and final report assessment, we found that blended teaching might provide an opportunity to stimulate greater motivation and interest in students as well as improve teaching effectiveness and learning outcomes of the course. This study describes how we conducted the CADD course during the COVID-19 period with the intention of providing a reference for other teachers to conduct similar courses.


Extracorporeal Membrane Oxygenation Cannulation Timing in the Pediatric Myocarditis Population: An Exploratory Analysis From the Extracorporeal Life Support Organization Registry.
Children presenting with acute myocarditis may experience rapid clinical deterioration requiring extracorporeal membrane oxygenation (ECMO); however, our understanding of best practices and timing of ECMO initiation are lacking. We explored the relationships between pre-cannulation factors and survival in this high-acuity patient population. Retrospective review of a large international registry. Primary outcome was survival to hospital discharge, stratified by incident cardiac arrest (CA) prior to ECMO and time to cannulation after intubation. The Extracorporeal Life Support Organization registry was queried for patients less than or equal to 18 years old receiving ECMO support for myocarditis between 2007 and 2018. Exclusion criteria included being nonindex runs, non-venoarterial ECMO or missing data points for main variables studied. None. Population characteristics and survival were compared using t test, Wilcoxon rank-sum test, or Fisher exact test. Multivariable logistic regression was used for significant factors in the unadjusted logistic regression. Among 506 index ECMO runs in pediatric patients with myocarditis, survival for the cohort was 72%, with no difference between early and late eras (2007-2012 vs 2013-2018; p = 0.69). Survivors demonstrated higher pre-ECMO pH levels as well as shorter intubation-to-cannulation (ITC) times (3 hr [interquartile range (IQR)], 1-14 hr vs 6 hr [IQR, 2-20 hr]; p = 0.021). CA occurred within 24 hours prior to ECMO cannulation, including extracorporeal cardiopulmonary resuscitation, in 54% of ECMO runs (n = 273). Accounting for the interaction between pre-ECMO CA occurrence and ITC time, longer ITC time remained associated with lower survival for patients who did not experience a CA prior to ECMO, with adjusted odds ratio of 0.09 (IQR, 0.02-0.40; p = 0.002) for ITC time greater than or equal to 18 hours. The results of this multicenter analysis of ECMO utilization and outcomes for pediatric myocarditis suggest that patients approaching ECMO cannulation who have not experienced CA may have better survival outcomes if cannulated onto ECMO early after intubation.


Evolution of Streptococcus pneumoniae and its close commensal relatives.
Streptococcus pneumoniae is a member of the Mitis group of streptococci which, according to 16S rRNA-sequence based phylogenetic reconstruction, includes 12 species. While other species of this group are considered prototypes of commensal bacteria, S. pneumoniae is among the most frequent microbial killers worldwide. Population genetic analysis of 118 strains, supported by demonstration of a distinct cell wall carbohydrate structure and competence pheromone sequence signature, shows that S. pneumoniae is one of several hundred evolutionary lineages forming a cluster separate from Streptococcus oralis and Streptococcus infantis. The remaining lineages of this distinct cluster are commensals previously collectively referred to as Streptococcus mitis and each represent separate species by traditional taxonomic standard. Virulence genes including the operon for capsule polysaccharide synthesis and genes encoding IgA1 protease, pneumolysin, and autolysin were randomly distributed among S. mitis lineages. Estimates of the evolutionary age of the lineages, the identical location of remnants of virulence genes in the genomes of commensal strains, the pattern of genome reductions, and the proportion of unique genes and their origin support the model that the entire cluster of S. pneumoniae, S. pseudopneumoniae, and S. mitis lineages evolved from pneumococcus-like bacteria presumably pathogenic to the common immediate ancestor of hominoids. During their adaptation to a commensal life style, most of the lineages gradually lost the majority of genes determining virulence and became genetically distinct due to sexual isolation in their respective hosts.


First Report of Dieback Disease on Cedars Caused by Diplodia seriata in China.
Cedars (Cedrus deodara (Roxb.) G. Don) are well known as evergreen ornamental trees widely used in horticulture in temperate climates. In March 2013, dieback symptoms were found on cedar trees in different locations (including the campus of Nanyang Normal University) in Nanyang (33°01' N, 112°29' E), a southwestern city of Henan Province, China. Characteristic symptoms included needle discoloration and defoliation, canker formation and gummosis on trunks and branches, browning and tissue necrosis under the bark as well as dieback of branches/trunks. Of 873 cedar trees investigated, 139 (16%) were symptomatic. A total of 21 Diplodia sp. isolates were obtained from 102 tissue pieces randomly sampled from the lesion margins of 31 affected trees with a conventional method for isolation of culturable fungal species from plant tissues. Monohyphal cultures were isolated from actively growing edges of colonies to purify the isolates. The purified isolates were grown on 2% water agar with sterilized stems of Foeniculum vulgare to induce their colonies to form pycnidia (4). Unmatured conidia were hyaline, aseptate, and turned light to dark brown with maturity. Mature conidia were aseptate (rarely uniseptate), ovoid with truncated or rounded base and obtuse apex, externally smooth, roughened on the inner surface, and 8 to 11 × 23 to 26 μm (n = 50). These morphological characteristics of the isolates agreed with those of Diplodia seriata (the anamorph of Botryosphaeria obtusa) (5). The rDNA-ITS sequences of two representative isolates (xs-01 and xs-06) were amplified with primers ITS1 and ITS4. PCR products were purified and ligated with PMD-19T vector for sequencing. The rDNA-ITS sequences were submitted to GenBank with accession nos. KJ463386 and KJ549774 for isolates xs-01 and xs-06, respectively, showing 100% identity with multiple isolates of D. seriata (HQ660463, KC461297, and KF535906). Koch's postulates were fulfilled in greenhouse tests on 2-year-old cedar plantlets inoculated by the two isolates. Ten plantlets were used for the inoculation tests for each of the isolates, and their trunks were wounded to a 2 mm depth with a sterilized cork borer (3 mm diameter). The wounds were inoculated by mycelial plugs cut from 7-day-old colonies grown on potato dextrose agar (PDA) plates and wrapped with Parafilm, and those inoculated with pure PDA plugs served as control. Inoculated plantlets were incubated in a greenhouse with alternating cycles of 14 h fluorescent light/10 h darkness under moist conditions for 30 to 60 days at 28°C. Nine of 20 inoculated plantlets developed needle discoloration and shoot blight symptoms similar to those observed on naturally infected cedar trees. The control plantlets remained symptomless during the incubation period. D. seriata cultures were constantly recovered from each diseased plantlet, indicating that the isolated D. seriata isolates were responsible for the disease. D. seriata has been reported as a phytopathogen causing dieback diseases worldwide on multiple woody plant species such as olive (4), mulberry (1), Pinus spp., and Picea glauca (2,3). To our knowledge, this is the first report of D. seriata causing dieback disease on cedars in China. References: (1) M. Arzanlou et al. Arch. Phytopathol. Plant Protect. 46:682, 2013. (2) T. Burgess et al. Appl. Environ. Microbiol. 67:354, 2001. (3) G. Hausner et al. Can. J. Plant Pathol. 21:256, 1999. (4) J. Kaliterna et al. Plant Dis. 96:290, 2012. (5) A. J. L. Phillips et al. Fungal Divers. 25:141, 2007.


Copy number variation of two separate regulatory regions upstream of SOX9 causes isolated 46,XY or 46,XX disorder of sex development.
SOX9 mutations cause the skeletal malformation syndrome campomelic dysplasia in combination with XY sex reversal. Studies in mice indicate that SOX9 acts as a testis-inducing transcription factor downstream of SRY, triggering Sertoli cell and testis differentiation. An SRY-dependent testis-specific enhancer for Sox9 has been identified only in mice. A previous study has implicated copy number variations (CNVs) of a 78 kb region 517-595 kb upstream of SOX9 in the aetiology of both 46,XY and 46,XX disorders of sex development (DSD). We wanted to better define this region for both disorders. By CNV analysis, we identified SOX9 upstream duplications in three cases of SRY-negative 46,XX DSD, which together with previously reported duplications define a 68 kb region, 516-584 kb upstream of SOX9, designated XXSR (XX sex reversal region). More importantly, we identified heterozygous deletions in four families with SRY-positive 46,XY DSD without skeletal phenotype, which define a 32.5 kb interval 607.1-639.6 kb upstream of SOX9, designated XY sex reversal region (XYSR). To localise the suspected testis-specific enhancer, XYSR subfragments were tested in cell transfection and transgenic experiments. While transgenic experiments remained inconclusive, a 1.9 kb SRY-responsive subfragment drove expression specifically in Sertoli-like cells. Our results indicate that isolated 46,XY and 46,XX DSD can be assigned to two separate regulatory regions, XYSR and XXSR, far upstream of SOX9. The 1.9 kb SRY-responsive subfragment from the XYSR might constitute the core of the Sertoli-cell enhancer of human SOX9, representing the so far missing link in the genetic cascade of male sex determination.


Is It Possible to Establish a Tumor-Suppressive Microenvironment With Glycine and Valine Supplement?
A tumorigenic microenvironment can give rise to neoplasm. A shift from this condition to a tumor-suppressive microenvironment is of significant benefit to susceptible individuals. The carbonyl groups of glycine and valine have long bond lengths, consequently generating potent affinities to divalent cations such as calcium. We hypothesize that the formation of insoluble and rigid calcium oxalate augmented by glycine and valine counteracts strong acids such as HCl chemically, thus reducing cancer risks. The anticancer effects of the 2 amino acids can be explained from a chemical and biochemical perspective. A tumor-suppressive microenvironment could be established via the modification of the proteome without genome editing at the DNA level.


Allosteric activation of vinculin by talin.
The talin-vinculin axis is a key mechanosensing component of cellular focal adhesions. How talin and vinculin respond to forces and regulate one another remains unclear. By combining single-molecule magnetic tweezers experiments, Molecular Dynamics simulations, actin-bundling assays, and adhesion assembly experiments in live cells, we here describe a two-ways allosteric network within vinculin as a regulator of the talin-vinculin interaction. We directly observe a maturation process of vinculin upon talin binding, which reinforces the binding to talin at a rate of 0.03 s-1. This allosteric transition can compete with force-induced dissociation of vinculin from talin only at forces up to 10 pN. Mimicking the allosteric activation by mutation yields a vinculin molecule that bundles actin and localizes to focal adhesions in a force-independent manner. Hence, the allosteric switch confines talin-vinculin interactions and focal adhesion build-up to intermediate force levels. The 'allosteric vinculin mutant' is a valuable molecular tool to further dissect the mechanical and biochemical signalling circuits at focal adhesions and elsewhere.


Dose-dependent effects of aluminum on osteocalcin synthesis in osteoblast-like ROS 17/2 cells in culture.
This in vitro study evaluates the effect of aluminum (Al3+) on osteocalcin, a small protein that is produced by the osteoblast. After stimulation with various doses of 1,25-dihydroxyvitamin D3 [1,25(OH)2D3; 10(-11) to 10(-9) M], osteocalcin was consistently lower in the culture medium of ROS 17/2 osteoblastic cells conditioned with 5 microM Al(3+)-saturated transferrin (AlTR) than in apotransferrin (ApoTR)-treated controls. In a second experiment, cultures were conditioned with various doses of AlTR or ApoTR (1.6-8.0 microM) and stimulated with 10(-9) M 1,25(OH)2D3. High doses of AlTR (4.8-8.0 microM) resulted in lower medium and unchanged intracellular content of osteocalcin than treatment with equal amounts of ApoTR. However, in the same experiment, lower doses of AlTR or ApoTR (1.6 and 3.2 microM) yielded different results, i.e., increased medium and intracellular contents of osteocalcin in the Al(3+)-treated cells. Expression of osteocalcin mRNA was not altered in cultures conditioned with low (1.6 microM) or high (8.0 microM) concentrations of AlTR or ApoTR. Similarly, no effect of Al3+ was observed on total protein content, the rate of total protein synthesis, and the degradation of secreted osteocalcin in cultures conditioned with various doses of AlTR or ApoTR. These findings suggest that AlTR affects osteocalcin synthesis in a specific manner, without concomitant effects on the rate of total protein synthesis or on the rate of degradation of osteocalcin. This effect is dose dependent, i.e., low doses of AlTR stimulate and high doses suppress osteocalcin synthesis and/or secretion, and it appears to be posttranscriptional, since the expression of osteocalcin mRNA is not affected.


Increased osteopontin plasma levels in multiple sclerosis patients correlate with bone-specific markers.
The pro-inflammatory cytokine osteopontin has been found to be highly expressed in multiple sclerosis lesions and plasma levels are increased during relapses in relapse-onset multiple sclerosis patients. The objective was to determine the relationship between osteopontin plasma and cerebrospinal fluid levels in relation to the immunoglobulin G index. In addition, osteopontin plasma levels were compared with osteopontin mRNA levels in peripheral blood mononuclear cells and bone-specific markers to analyse whether osteopontin may be peripherally produced. Osteopontin and bone-specific markers were determined in paired plasma-cerebrospinal fluid samples and serum samples of relapse-onset multiple sclerosis patients (n = 36), respectively. Osteopontin mRNA levels were determined by quantitative polymerase chain reaction analysis. Compared to healthy controls (n = 20), plasma osteopontin levels were significantly increased in relapsing-remitting multiple sclerosis patients and correlated (r = 0.43, p = 0.013) with the immunoglobulin G index. In contrast, cerebrospinal fluid osteopontin levels correlated neither with plasma osteopontin in paired samples nor with the immunoglobulin G index. Since osteopontin mRNA levels in peripheral blood mononuclear cells of relapsing-remitting multiple sclerosis patients did not correlate with osteopontin plasma levels, peripheral blood mononuclear cells might not be the major source for the increased osteopontin plasma levels. Osteopontin plasma levels correlated (r = 0.42, p = 0.035) with the bone-specific degradation product C-telopeptide of type-1 collagen. In addition, another immunomodulatory molecule involved in bone metabolism, 25-OH vitamin D, correlated negatively (r = -0.359, p = 0.048) with the immunoglobulin G index. This study suggests that bone-related molecules like osteopontin and vitamin D with important immunomodulatory functions are related to the immunoglobulin G index in relapsing-remitting multiple sclerosis patients.


A model clinic for the initial evaluation and establishment of primary care for persons infected with human immunodeficiency virus.
The human immunodeficiency virus (HIV)-infected patients on initial presentation to primary medical care may have extensive problems relating to medical, substance abuse, psychiatric, and social issues. We examined a model for the initiation of primary medical care for patients infected with HIV at a public hospital and presented the clinical, educational, and research benefits associated with such a clinic. Eight hundred forty-five consecutive HIV-infected outpatients without primary care, who presented to a municipal hospital HIV intake clinic, the Diagnostic Evaluation Unit, between February 1, 1990 and August 31, 1993, received a multidisciplinary assessment and a facilitated access to medical services. The performance of standardized initial clinical evaluation and adherence to primary care referral were examined. During the most recent study year, more than 90% of patients presenting to the clinic completed the intake process and 95% were seen at the site to which they were referred for primary medical care. Tests for CD4 lymphocytes, syphilis, hepatitis B, and tuberculosis were obtained, pneumococcal vaccinations were administered, and social service assessments were performed in 92% to 98% of patients completing the intake. The clinical setting was a site for involvement in research protocols and provided a focused educational experience concerning outpatient HIV management for medical students and residents. Establishment of a clinic dedicated to the initial evaluation of HIV-infected persons in a municipal hospital successfully evaluated and linked patients to primary care providers. The clinic structure enabled the standardized performance of appropriate laboratory tests and vaccinations and provided unique educational and research opportunities. We encourage other health care settings to consider the development of similar models for the initiation of medical care for persons infected with HIV.


Synthesis and endothelin receptors binding affinity of new 1,3,5- substituted pyrrole-2-carboxylic acid derivatives.
The interest of researchers for ligands of the endothelin receptors ETA and ETB is due to their extensive therapeutic potential. In particular, receptor antagonists are useful in a number of diseases such as pulmonary hypertension, acute myocardial infarction, congestive heart failure, renal failure, and atherosclerosis. In the context of our research program aimed to the development of new endothelin receptor ligands, in this paper we describe the synthesis and structure- activity relationships of a new series of 1,3,5-substituted pyrrole-2-carboxylic acid derivatives 27-40 possessing the structural features for ET receptors binding. New synthesized compounds were tested on ETA and ETB receptors stably expressed in CHO cells and some of them showed interesting affinity and selectivity towards ETA receptors.


Characteristics of Dutch and Swiss primary care COPD patients - baseline data of the ICE COLD ERIC study.
INTERNATIONAL COLLABORATIVE EFFORT ON CHRONIC OBSTRUCTIVE LUNG DISEASE: Exacerbation Risk Index Cohorts (ICE COLD ERIC) is a prospective cohort study with chronic obstructive pulmonary disease (COPD) patients from Switzerland and The Netherlands designed to develop and validate practical COPD risk indices that predict the clinical course of COPD patients in primary care. This paper describes the characteristics of the cohorts at baseline. Standardized assessments included lung function, patient history, self-administered questionnaires, exercise capacity, and a venous blood sample for analysis of biomarkers and genetics. A total of 260 Dutch and 151 Swiss patients were included. Median age was 66 years, 57% were male, 38% were current smokers, 55% were former smokers, and 76% had at least one and 40% had two or more comorbidities with cardiovascular disease being the most prevalent one. The use of any pulmonary and cardiovascular drugs was 84% and 66%, respectively. Although lung function results (median forced expiratory volume in 1 second [FEV(1)] was 59% of predicted) were similar across the two cohorts, Swiss patients reported better COPD-specific health-related quality of life (Chronic Respiratory Questionnaire) and had higher exercise capacity. COPD patients in the ICE COLD ERIC study represent a wide range of disease severities and the prevalence of multimorbidity is high. The rich variation in these primary care cohorts offers good opportunities to learn more about the clinical course of COPD.


Glucose Phosphotransferase System Modulates Pyruvate Metabolism, Bacterial Fitness, and Microbial Ecology in Oral Streptococci.
Spontaneous mutants with defects in the primary glucose phosphotransferase permease (manLMNO) of Streptococcus sanguinis SK36 showed enhanced fitness at low pH. Transcriptomics and metabolomics with a manL deletion mutant (SK36/manL) revealed redirection of pyruvate to production of acetate and formate, rather than lactate. These observations were consistent with measurements of decreased lactic acid accumulation and increased excretion of acetate, formate, pyruvate, and H2O2. Genes showing increased expression in SK36/manL included those encoding carbohydrate transporters, extracellular glycosidases, intracellular polysaccharide metabolism, and arginine deiminase and pathways for metabolism of acetoin, ethanolamine, ascorbate, and formate, along with genes required for membrane biosynthesis and adhesion. Streptococcus mutans UA159 persisted much better in biofilm cocultures with SK36/manL than with SK36, an effect that was further enhanced by culturing the biofilms anaerobically but dampened by adding arginine to the medium. We posited that the enhanced persistence of S. mutans with SK36/manL was in part due to excess excretion of pyruvate by the latter, as addition of pyruvate to S. mutans-S. sanguinis cocultures increased the proportions of UA159 in the biofilms. Reducing the buffer capacity or increasing the concentration of glucose benefited UA159 when cocultured with SK36, but not with SK36/manL, likely due to the altered metabolism and enhanced acid tolerance of the mutant. When manL was deleted in S. mutans or Streptococcus gordonii, the mutants presented altered fitness characteristics. Our study demonstrated that phosphotransferase system (PTS)-dependent modulation of central metabolism can profoundly affect streptococcal fitness and metabolic interactions, revealing another dimension in commensal-pathogen relationships influencing dental caries development. IMPORTANCE Dental caries is underpinned by a dysbiotic microbiome and increased acid production. As beneficial bacteria that can antagonize oral pathobionts, oral streptococci such as S. sanguinis and S. gordonii can ferment many carbohydrates, despite their relative sensitivity to low pH. We characterized the molecular basis for why mutants of glucose transporter ManLMNO of S. sanguinis showed enhanced production of hydrogen peroxide and ammonia and improved persistence under acidic conditions. A metabolic shift involving more than 300 genes required for carbohydrate transport, energy production, and envelope biogenesis was observed. Significantly, manL mutants engineered in three different oral streptococci displayed altered capacities for acid production and interspecies antagonism, highlighting the potential for targeting the glucose-PTS to modulate the pathogenicity of oral biofilms.


The use of the finasteride-adjusted Prostate Cancer Prevention Trial Prostate Cancer Risk Calculator in a Mexican referral population: a validation study.
To perform the first validation study of the finasteride-adjusted Prostate Cancer Prevention Trial Prostate Cancer Risk Calculator (finPCPTRC) in a contemporary referral population in Mexico. 837 patients referred to the Instituto Nacional de Cancerología, Mexico City, Mexico, between 2005 and 2009 were used to validate the finPCPTRC by examining various measures of discrimination and calibration. Net benefit curve analysis was used to gain insight into the use of the finPCPTRC for clinical decisions. Prostate cancer (PCa) incidence (72.8%) was high in this Mexican referral cohort and 45.7% of men who were diagnosed with PCa had high-grade lesions (HGPCa, Gleason score >6). 1.3% of the patients were taking finasteride. The finPCPTRC was a superior diagnostic tool compared to prostate-specific antigen alone when discriminating patients with PCa from those without PCa (AUC = 0.784 vs. AUC = 0.687, p < 0.001) and when discriminating patients with HGPCa from those without HGPCa (AUC = 0.768 vs. AUC = 0.739, p < 0.001). The finPCPTRC underestimated the risk of PCa but overestimated the risk of HGPCa (both p < 0.001). Compared with other strategies to opt for biopsy, the net benefit would be larger with utilization of the finPCPTRC for patients accepting higher risks of HGPCa. Rates of biopsy-detectable PCa and HGPCa were high and 1.3% of this referral cohort in Mexico was taking finasteride. The risks of PCa or HGPCa calculated by the finPCPTRC were not well calibrated for this referral Mexican population and new clinical diagnostic tools are needed.


The accuracy of digital templating in uncemented total hip arthroplasty.
We analyzed the reproducibility and reliability of computer templating in primary uncemented total hip arthroplasties as compared to standard onlay templating techniques with hardcopy radiographs from a digital source. Digital templating showed good intraobserver and interoberserver reliability with intraclass correlation coefficient values greater than 0.7. Using computer templating, prediction of sizing to within 1 size was 85% accurate for femoral stem sizing and 80% accurate for acetabular sizing. Using onlay templating, prediction of sizing to within 1 size was 85% accurate for femoral sizing and 60% accurate for acetabular sizing. We conclude that the introduction of digital templating has significant benefits in preoperative planning for total hip arthroplasty over onlay templating.


Changes in genome content generated via segregation of non-allelic homologs.
A careful analysis of two maize recombinant inbred lines (RILs) relative to their inbred parents revealed the presence of several hundred apparently de novo copy number variants (CNVs). These changes in genome content were validated via both PCR and whole exome-array capture-and-sequencing experiments. One hundred and eighty-five genomic regions, which overlap with 38 high-confidence genes, exhibited apparently de novo copy number variation (CNV) in these two RILs and in many instances the same apparently de novo CNV events were observed in multiple RILs. Further analyses revealed that these recurrent apparently de novo CNVs were caused by segregation of single-copy homologous sequences that are located in non-allelic positions in the two parental inbred lines. F(1) individuals derived from these inbred lines will be hemizygous for each of these non-allelic homologs but RIL genotypes will contain these sequences at zero, one or two genomic loci. Hence, the segregation of non-allelic homologs may contribute to transgressive segregation. Indeed, statistical associations between phenotypic quantitative trait loci and genomic losses were observed for two of 14 tested pairs of non-allelic homologs.


Panagrolaimus superbus tolerates hypoxia within Gallium metal cage: implications for the understanding of the phenomenon of anhydrobiosis.
Panagrolaimus superbus nematodes are able to tolerate desiccation by entering into a peculiar state of suspended animation known as anhydrobiosis. When desiccated, anhydrobiotic organisms are also able to tolerate other physical stresses, as high and low levels of temperature and pressure. Here, we decided to investigate the tolerance of desiccated P. superbus to an unprecedented double stress - hypoxia within 99.99% Gallium (Ga) metal cage. The authors observed that regardless of the external relative humidity, desiccated P. superbus tolerated 7 d confined within the metal cage, displaying no negative effects on its survival and population growth rates over 40 d. The results evidence that anhydrobiosis also renders nematodes tolerant to otherwise lethal concentrations of Ga, in an oxygen-poor environment; thus, expanding its polyextremotolerance profile. Panagrolaimus superbus nematodes are able to tolerate desiccation by entering into a peculiar state of suspended animation known as anhydrobiosis. When desiccated, anhydrobiotic organisms are also able to tolerate other physical stresses, as high and low levels of temperature and pressure. Here, we decided to investigate the tolerance of desiccated P. superbus to an unprecedented double stress – hypoxia within 99.99% Gallium (Ga) metal cage. The authors observed that regardless of the external relative humidity, desiccated P. superbus tolerated 7 d confined within the metal cage, displaying no negative effects on its survival and population growth rates over 40 d. The results evidence that anhydrobiosis also renders nematodes tolerant to otherwise lethal concentrations of Ga, in an oxygen-poor environment; thus, expanding its polyextremotolerance profile.


The significance of fluorinated compound chain length, treatment technology, and influent composition on per- and polyfluoroalkyl substances removal in worldwide wastewater treatment plants.
Wastewater treatment plants (WWTPs) are deemed major conveyors and point sources of per- and polyfluoroalkyl substances (PFAS) to the environment. This statistical meta-analysis of existing literature from the past 15 years focused on the significance of treatment type for PFAS removal efficiencies and the influence of PFAS sources (domestic vs. industrial) on their removal. Different sampling events, WWTPs across the world, different treatment technologies, configurations, and processes, as well as different PFAS classes and compounds were considered. This study evaluated 13 PFAS analyzed predominantly in 161 WWTPs across the world. The statistical test results revealed that these 13 frequently detected and reported PFAS can be divided into four groups based on their behavior during wastewater treatment, namely (1) C6-10 perfluorocarboxylic acids (PFCAs), (2) C4,5,11,12 PFCAs, (3) C4,6,8 perfluoroalkane sulfonic acids (PFSAs), and (4) C10 PFSA. In this study, biological treatments such as (1) membrane bioreactors, (2) combination of two or more biological treatments, and (3) biofilm processes revealed the highest PFAS removals, although the addition of a tertiary treatment actually had a nonbeneficial effect on PFAS removal. Moreover, a strong statistical correlation was observed between industrial wastewater sources and the presence of high influent PFAS concentrations in the receiving WWTPs. This indicates that industrial sources were the main contributors of the PFAS load in the analyzed WWTPs. Integr Environ Assess Manag 2023;00:1-11. © 2023 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).


Feasibility and safety of planned early discharge following laparotomy in gynecologic oncology with enhanced recovery protocol including opioid-sparing anesthesia.
This study aims to evaluate the feasibility and safety of planned postoperative day 1 discharge (PPOD1) among patients who undergo laparotomy (XL) in the department of gynecology oncology utilizing a modified enhanced recovery after surgery (ERAS) protocol including opioid-sparing anesthesia (OSA) and defined discharge criteria. Patients undergoing XL and minimally invasive surgery (MIS) were enrolled in this prospective, observational cohort study after the departmental implementation of a modified ERAS protocol. The primary outcome was quality of life (QoL) using SF36, PROMIS GI, and ICIQ-FLUTS at baseline and 2- and 6-week postoperative visits. Statistical significance was assessed using the two-tailed Student's t-test and non-parametric Mann-Whitney two-sample test. Of the 141 subjects, no significant demographic differences were observed between the XL group and the MIS group. The majority of subjects, 84.7% (61), in the XL group had gynecologic malignancy [vs. MIS group; 21 (29.2%), p < 0.001]. All patients tolerated OSA. The XL group required higher intraoperative opioids [7.1 ± 9.2 morphine milligram equivalents (MME) vs. 3.9 ± 6.9 MME, p = 0.02] and longer surgical time (114.2 ± 41 min vs. 96.8 ± 32.1 min, p = 0.006). No significant difference was noted in the opioid requirements at the immediate postoperative phase and the rest of the postoperative day (POD) 0 or POD 1. In the XL group, 69 patients (73.6%) were successfully discharged home on POD1. There was no increase in the PROMIS score at 2 and 6 weeks compared to the preoperative phase. The readmission rates within 30 days after surgery (XL 4.2% vs. MIS 1.4%, p = 0.62), rates of surgical site infection (XL 0% vs. MIS 2.8%, p = 0.24), and mean number of post-discharge phone calls (0 vs. 0, p = 0.41) were comparable between the two groups. Although QoL scores were significantly lower than baseline in four of the nine QoL domains at 2 weeks post-laparotomy, all except physical health recovered by the 6-week time point. PPOD1 is a safe and feasible strategy for XL performed in the gynecologic oncology department. PPOD1 did not increase opioid requirements, readmission rates compared to MIS, and patient-reported constipation and nausea/vomiting compared to the preoperative phase.


Investigations on Piper betle grown in Sri Lanka.
Piper betle is an economically important plant cultivated in Sri Lanka. Although more than 12 cultivars of betel are reported in Sri Lanka, very few scientific investigations have been carried out on them. Studies on the chemical constituents indicated that safrole is the major constituent, followed by chavibitol acetate, in the essential oil of common betel leaves of Sri Lanka. Investigations on the bioactivities of P. betle revealed the presence of antimicrobial, insecticidal, antioxidant, antinociceptive, antidiabetic and gastroprotective activities. In addition, P. betle was found to be safe in terms of hepatotoxicity, renotoxicity, hematotoxicity, gross morphology, weights of organs, stress or aversive behaviors in rats. The above findings indicate the vast potential of P. betle yet to be harnessed for the benefit of mankind and the betel industry of Sri Lanka.


Effect of cimetidine on 24-hour intragastric acidity in normal subjects.
The effect of H2-receptor blockade on intragastric acidity was studied in nine normal males. The pH of their gastric contents was measured at hourly daytime and two hourly nighttime intervals for 48 hours. The subjects ate identical meals, drank identical volumes of fluid, and smoked the same number of cigarettes during the two study days. Their physical activity was unrestricted in a ward environment. Blood cimetidine and plasma gastrin were measured in serial blood samples. The nine subjects were treated in random sequence with cimetidine 0-8-1-0 g on one day and placebo capsules on the other. The drug was given in four divided doses: four subjects received it before, and five after, the three main meals. All took the fourth dose at bedtime. Replicate studies in an additional subject given placebo on both study days showed good reproducibility (r=0-80, P less than 0-01). Cimetidine therapy decreased intragastric acidity in all nine subjects. The decrease was similar in the two groups taking the drug before or after meals, mean 24 h intragastric hydrogen ion activity being lowered by 70 and 72% respectively. Nocturnal anacidity was recorded in only two of 45 samples. Administration of cimetidine before meals produced earlier and higher drug blood levels than post-prandial medication, but when it was taken after food the blood levels were highest at the time when the buffer capacity of the food was waning. Blood concentrations of cimetidine exceeded the secretory IC50 level for most of the time between doses. The results show that cimetidine 0-8-1-0 g/day in four divided doses produces a striking and consistent decrease of intragastric acidity. Although variation in the timing of the dose in relation to meals did not affect the decrease of acidity, the absorption data suggest that patients should take the drug after meals.


Non-lethal exposure to H2O2 boosts bacterial survival and evolvability against oxidative stress.
Unicellular organisms have the prevalent challenge to survive under oxidative stress of reactive oxygen species (ROS) such as hydrogen peroxide (H2O2). ROS are present as by-products of photosynthesis and aerobic respiration. These reactive species are even employed by multicellular organisms as potent weapons against microbes. Although bacterial defences against lethal and sub-lethal oxidative stress have been studied in model bacteria, the role of fluctuating H2O2 concentrations remains unexplored. It is known that sub-lethal exposure of Escherichia coli to H2O2 results in enhanced survival upon subsequent exposure. Here we investigate the priming response to H2O2 at physiological concentrations. The basis and the duration of the response (memory) were also determined by time-lapse quantitative proteomics. We found that a low level of H2O2 induced several scavenging enzymes showing a long half-life, subsequently protecting cells from future exposure. We then asked if the phenotypic resistance against H2O2 alters the evolution of resistance against oxygen stress. Experimental evolution of H2O2 resistance revealed faster evolution and higher levels of resistance in primed cells. Several mutations were found to be associated with resistance in evolved populations affecting different loci but, counterintuitively, none of them was directly associated with scavenging systems. Our results have important implications for host colonisation and infections where microbes often encounter reactive oxygen species in gradients.


Synergistic cytotoxic activity of recombinant TRAIL plus the non-genotoxic activator of the p53 pathway nutlin-3 in acute myeloid leukemia cells.
To potentiate the response of acute myeloid leukemia (AML) to TRAIL cytotoxicity, we have adopted a strategy of combining nutlin-3, a potent non-genotoxic activator of the p53 pathway, with recombinant TRAIL. The rationale for using such a combination was that deletions and/or mutations of the p53 gene occur in only 5-10% of AML and that TRAIL and nutlin-3 activate the extrinsic and intrinsic pathways of apoptosis, respectively. TRAIL induced a rapid increase of apoptosis when added to OCI M4-type and MOLM M5-type AML cells, carrying a wild-type p53, as well as to NB4 M3-type AML, carrying a mutated p53. On the other hand, the small molecule activator of the p53 pathway nutlin-3 induced p53 accumulation, cell cycle arrest and a slow progressive increase of apoptosis in OCI and MOLM but not in NB4. Of note, nutlin-3 up-regulated the surface expression of TRAIL-R2 and synergized with TRAIL in inducing apoptosis in OCI and MOLM as well as in primary M4-type and M5-type AML blasts, but not in NB4 cells. Moreover, while nutlin-3 up-regulated the expression of cyclin dependent kinase inhibitor p21, a p53-target gene mediating cell cycle block and showing anti-apoptotic activity, the simultaneous addition of TRAIL plus nutlin-3 induced the caspase-dependent cleavage of p21. The relevance of p21 down-regulation for sensitizing AML cells to apoptosis was underscored in knocking-down experiments with small interfering RNAs. Our data suggest that the combined treatment of nutlin-3 plus TRAIL might offer a novel therapeutic strategy for AML.


The inhibitory action of quercetin on lipopolysaccharide-induced nitric oxide production in RAW 264.7 macrophage cells.
The effect of quercetin on lipopolysaccharide (LPS)-induced nitric oxide (NO) production was studied. Quercetin pretreatment significantly inhibited NO production in an LPS-stimulated RAW 264.7 murine macrophage cell line. Post-treatment with quercetin partially inhibited NO production. The inhibitory action of quercetin was due to neither the cytotoxic action nor altered LPS binding. The expression of inducible-type NO synthase (iNOS) was markedly down-regulated by quercetin. Quercetin suppressed the release of free nuclear factor (NF)-kappaB by preventing degradation of IkappaB-alpha and IkappaB-beta. Moreover, quercetin blocked the phosphorylation of extracellular signal regulated kinase 1/2 (Erk 1/2), p38, and c-Jun NH2-terminal kinase/stress-activated protein kinase (JNK/SAPK) and, further, the activity of tyrosine kinases in LPS-stimulated RAW cells. Quercetin also inhibited interferon (IFN)-gamma-induced NO production. Taken together, these results indicate that the inhibitory action of quercetin on NO production in LPS- and/or IFN-gamma-stimulated macrophages might be due to abrogation of iNOS protein induction by impairment of a series of intracellular signal pathways.


Corynebacterium kroppenstedtii: a challenging culprit in breast abscesses and granulomatous mastitis.
Corynebacterium kroppenstedtii is a difficult pathogen associated with granulomatous mastitis and recurrent breast abscesses. Despite over a dozen studies, there is no guidance on surgical interventions, steroid use, or dosing or duration of antibiotic treatment. Initially seen in predominantly Maori and Pacific Islander multiparous, postlactating women, C. kroppenstedtii breast infection has since been reported throughout the world, including in nulliparous women. Additionally, emerging data suggest that hyperprolactinemia is a modifiable risk factor for these infections. This article reviews a patient case and data from 87 other cases to compile current best practices for diagnosis, treatment, and monitoring, and provides areas for future study. In cases of granulomatous mastitis and breast abscess, especially if recurrent, infection with C. kroppenstedtii should be considered. Microbiologists should be alerted to the specialized growth conditions and tools needed for appropriate culturing, identification, and antibiotic susceptibility testing. Clinicians should utilize a multimodal approach with surgical and antibiotic treatment to maximize clinical cure and reduce recurrence.


The use of Lean and Six Sigma methodologies in surgery: a systematic review.
Lean and Six Sigma are improvement methodologies developed in the manufacturing industry and have been applied to healthcare settings since the 1990 s. They use a systematic and reproducible approach to provide Quality Improvement (QI), with a flexible process that can be applied to a range of outcomes across different patient groups. This review assesses the literature with regard to the use and utility of Lean and Six Sigma methodologies in surgery. MEDLINE, Embase, PsycINFO, Allied and Complementary Medicine Database, British Nursing Index, Cumulative Index to Nursing and Allied Health Literature, Health Business Elite and the Health Management Information Consortium were searched in January 2014. Experimental studies were included if they assessed the use of Lean or Six Sigma on the ability to improve specified outcomes in surgical patients. Of the 124 studies returned, 23 were suitable for inclusion with 11 assessing Lean, 6 Six Sigma and 6 Lean Six Sigma. The broad range of outcomes can be collated into six common aims: to optimise outpatient efficiency, to improve operating theatre efficiency, to decrease operative complications, to reduce ward-based harms, to reduce mortality and to limit unnecessary cost and length of stay. The majority of studies (88%) demonstrate improvement; however high levels of systematic bias and imprecision were evident. Lean and Six Sigma QI methodologies have the potential to produce clinically significant improvement for surgical patients. However there is a need to conduct high-quality studies with low risk of systematic bias in order to further understand their role.


Hypoxia enhances IPF mesenchymal progenitor cell fibrogenicity via the lactate/GPR81/HIF1α pathway.
Hypoxia is a sentinel feature of idiopathic pulmonary fibrosis (IPF). The IPF microenvironment contains high lactate levels, and hypoxia enhances cellular lactate production. Lactate, acting through the GPR81 lactate receptor, serves as a signal molecule regulating cellular processes. We previously identified intrinsically fibrogenic mesenchymal progenitor cells (MPCs) that drive fibrosis in the lungs of patients with IPF. However, whether hypoxia enhances IPF MPC fibrogenicity is unclear. We hypothesized that hypoxia increases IPF MPC fibrogenicity via lactate and its cognate receptor GPR81. Here we show that hypoxia promotes IPF MPC self-renewal. The mechanism involves hypoxia-mediated enhancement of LDHA function and lactate production and release. Hypoxia also increases HIF1α levels, and this increase in turn augments the expression of GPR81. Exogenous lactate operating through GPR81 promotes IPF MPC self-renewal. IHC analysis of IPF lung tissue demonstrates IPF MPCs expressing GPR81 and hypoxic markers on the periphery of the fibroblastic focus. We show that hypoxia enhances IPF MPC fibrogenicity in vivo. We demonstrate that knockdown of GPR81 inhibits hypoxia-induced IPF MPC self-renewal in vitro and attenuates hypoxia-induced IPF MPC fibrogenicity in vivo. Our data demonstrate that hypoxia creates a feed-forward loop that augments IPF MPC fibrogenicity via the lactate/GPR81/HIF1α pathway.


[Typology of defunctioning colostomy and state of art in the treatment of bowel emergencies].
A trend toward avoidance of a defunctioning colostomy at emergency large-bowel surgery has been placed in recent years. The surgical management of patients with acute colonic disease has been evolving from multiple to single operations with a reduced use of colostomy. One hundred four consecutive non-selected patients underwent surgery for left-sided large bowel emergencies between 1980-2003. Defunctioning colostomy was performed in 10 out of 58 resection-anastomosis procedures. Thirty-seven patients underwent Hartmann procedure, 9 received only diverting colostomy. Postoperative morbidity was 28.8%. Postoperative mortality 8.2%. Anastomotic leak occurred in 1 and 6 patients with and without defunctioning colostomy respectively. Four out of the 6 patients without colostomy needed reintervention, while patient with covering colostomy underwent conservative treatment. Six (10.5%) out 56 patients with colostomy experienced major stoma related complications and underwent reintervention. Although there is general acceptance of one-stage surgery for right-sided colon emergencies, the surgical management of left-side large bowel obstruction and peritonitis remains controversal. Risk of anastomotic dehiscence associated with large-bowel anastomosis in unfavourable circumstance must be balanced against the high complications and low closure rates of a temporary colostomy. Primary resection and anastomosis without diverting colostomy for left-sided acute obstruction and peritonitis may be performed in selected patients. Diffuse purulent and faecal peritonitis are contraindications to one-stage surgery being necessary a two- stage procedure with loop or end colostomy. Colostomy remain a valid surgical option when high risk of dehiscence is suspected.


Levo-tetrahydropalmatine decreases ethanol drinking and antagonizes dopamine D2 receptor-mediated signaling in the mouse dorsal striatum.
An herb derived compound, levo-tetrahydropalmatine (L-THP), attenuates self-administration of cocaine and opiates in rodents. Since L-THP mainly antagonizes dopamine D2 receptors (D2R) in the brain, it is likely to regulate other addictive behaviors as well. Here, we examined whether L-THP regulates ethanol drinking in C57BL/6J mice using a two-bottle choice drinking experiment. L-THP treated mice consumed less ethanol compared to vehicle-treated mice during the 15% ethanol drinking session while water consumption remained similar between each group. We then examined the molecular basis underlying the pharmacological effect of L-THP in mice. Our results indicated that a single injection of L-THP increased active phosphorylated forms of PKA, AKT and ERK in the caudate-putamen (CPu), but not in the nucleus accumbens (NAc), of alcohol naïve mice. Interestingly, we found that systematic treatment with L-THP for 4 consecutive days while mice were drinking 15% ethanol increased pPKA levels in the CPu, but not in the NAc. In contrast to the effect of acute L-THP treatment, no differences were detected for pAKT or pERK in either striatal regions. Together, our findings suggest that reduction of ethanol drinking by L-THP treatment is possibly correlated with D2R-mediated PKA signaling in the CPu.


Clinical experience with combination BRAF/MEK inhibitors for melanoma with brain metastases: a real-life multicenter study.
BRAF and MEK kinase inhibitors can be highly effective in treating BRAF-mutant melanomas, but their safety and activity in patients with active/symptomatic brain metastases are unclear. We sought to shed light on this open clinical question. We conducted a multicenter retrospective study on real-life patients with melanoma and active brain metastases treated with combination BRAF/MEK inhibitors. A total of 65 patients were included (38 men and 27 women; median age: 49 years). Of them, 53 patients received dabrafenib/trametinib, 10 received vemurafenib/cobimetinib, one received encorafenib/binimetinib, and one received vemurafenib/trametinib. We did not observe any unexpected treatment-related safety signals in our cohort. Overall, 17 patients continued on therapy through the cutoff date. After initiation of therapy, steroid dose could be decreased in 22 of 33 patients (11 tapered off entirely), anticonvulsants were stopped in four of 21, and narcotics were stopped in four of 12. Median progression-free survival from the start of therapy was 5.3 months (95% confidence interval: 3.6-6.1), and median overall survival was 9.5 months (95% confidence interval: 7.7-13.5). A total of 20 patients were surviving at the cutoff date. Univariate analysis of age, sex, ulceration status, thickness, stage, location, or lactate dehydrogenase did not reveal significant predictors of progression-free survival or overall survival within our cohort, but multivariate analysis suggested that older age, lower risk location of original lesion, and nodular melanoma are poor prognostic indicators. Combination therapy with BRAF/MEK inhibitors is a viable treatment option for patients with BRAF-mutant melanoma and brain metastases, but further studies should help to define the optimal treatment approach in this population.


Chemical stability of extra-virgin olive oil added with oregano essential oil.
Extra virgin olive oil is highly consumed and well known for its nutritional and health benefits. However, it is fatty food highly susceptible to lipid oxidation. The objective of this study was to evaluate the preserving effect of oregano (Origanum vulgare L. spp vulgare called "oregano compacto") essential oil on physical and chemical properties in extra virgin olive oil during storage. Oregano essential oil composition was analyzed by GC-MS. This essential oil was added into extra virgin olive oil at 0.05%. The samples were stored in 3 different conditions: darkness, light exposure, and temperature (60 °C). Chemical indicators of lipid oxidation (peroxide value, p-anisidine value, conjugated dienes, free fatty acidity, and carotenoid and chlorophyll contents) were measured. High content in carvomenthol (22.52%), terpinolene (19.77%), thymol (13.51%), and γ-terpinene (10.30%) were detected in oregano essential oil. Olive oil samples without oregano essential oil stored at 60 °C and exposure at artificial light had the highest peroxide values during storage. Higher p-anisidine and K232 values after day 7 of storage were detected in temperature, darkness, and light exposure treatments. Light treatment was the main factor that degraded chlorophyll causing loss of color. The highest chlorophyll content (3.87 mg/kg) was observed in olive oil with essential oil at the end of storage. In general, olive oil samples added with oregano essential oil had lower peroxide, conjugated dienes, and p-anisidine values and higher chlorophyll and carotenoid contents during storage. Oregano essential oil retards lipid oxidation process in olive oil prolonging its shelf life. Oregano essential oil was and is used with the purpose of flavoring and aromatizing food. This essential oil due to its composition has shown antioxidant activity. Synthetic antioxidants such as butylated hydroxytoluene (BHT) and butylated hydroxyanisole (BHA) are thought to be promoters of carcinogenesis. Extra virgin olive oil is widely consumed because of its nutritional benefits and sensory properties which are very important to be preserved in the product. In this study, the oregano essential oil showed remarkable antioxidant activity in olive oil. Therefore, this essential oil could be considered for the industry as natural antioxidant not only to be used in olive oil but also in other fatty food products to substitute synthetic ones.


Mass spectrometry-based quantitative proteomic profiling.
Quantitative proteomics involves the identification and quantitation of protein components in various biological systems. Stable isotope labelling technology, by both metabolic and chemical methods, has been the most commonly used approach for global proteome-wide profiling. Recently, its capability has been extended from labelled pairs to multiple labels, allowing for the simultaneous quantification of multiplex samples. The ion intensity-based quantitative approach has progressively gained more popularity as mass spectrometry performance has improved significantly. Although some success has been reported, it remains difficult comprehensively to characterise the global proteome, due to its enormous complexity and dynamic range. The use of sub-proteome fractionation techniques permits a simplification of the proteome and provides a practical step towards the ultimate dissection of the entire proteome. Further development of the technology for targeting sub-proteomes on a functional basis - such as selecting proteins with differential expression profiles from mass spectrometric analyses, for further mass spectrometric sequencing in an intelligent manner--is expected in the near future.


Human immunodeficiency virus infection in a Dublin general practice.
A group general practice in Dublin's inner city has had extensive experience of intravenous drug users since the late 1970s. Since 1985 a total of 54 human immunodeficiency virus (HIV) seropositive patients have attended the practice, of whom 48 are intravenous drug users, four are the children of drug users and two have been infected through sexual contacts. Three patients have developed the acquired immune deficiency syndrome and at least eight have symptomatic HIV disease. Sixty per cent of Ireland's seropositive population have been infected through intravenous drug abuse but nationally only 16% of all intravenous drug users tested are seropositive; in the study practice, however, at least 35% (48/137) of known intravenous drug users are seropositive.


Measuring induction times and crystal nucleation rates.
A large variation is observed in induction times measured under equal conditions in 1 ml solutions. Ruling out experimental errors, this variation originates from the nucleation process. The induction time distribution is explained by the stochastic nature of nucleation if the number of nuclei formed is approaching 1 per vial. Accurate heterogeneous crystal nucleation rates were determined from the induction time distributions on a 1 ml scale for racemic diprophylline in two solvents. The difference in nucleation behaviour in the two solvents originates from the energy barrier for nucleation, which is much higher in the solvent in which induction times are much longer. In addition the pre-exponential factor for the crystal nucleation rate in both solvents is rather low compared to predictions using Classical Nucleation Theory. Unfortunately, concentration and surface characteristics of the effective heterogeneous particles are not known which clouds a further molecular interpretation.


Rapid Response and Slow Recovery of the H3K4me3 Epigenomic Marker in the Liver after Light-mediated Phase Advances of the Circadian Clock.
Mammalian tissues display circadian rhythms in transcription, translation, and histone modifications. Here we asked how an advance of the light-dark cycle alters daily rhythms in the liver epigenome at the H3K4me3 (trimethylation of lysine 4 on histone 3) modification, which is found at active and poised gene promoters. H3K4me3 levels were first measured at 4 time points (zeitgeber time [ZT] 3, 8, 15, and 20) during a normal 12L:12D light-dark cycle. Peak levels were observed during the early dark phase at ZT15 and dropped to low levels around lights-on (ZT0) between ZT20 and ZT3. A 6-h phase advance at ZT18 (new lights-on after only 6 h of darkness) led to a transient extension of peak H3K4me3 levels. Although locomotor activity reentrained within a week after the phase advance, H3K4me3 rhythms failed to do so, with peak levels remaining in the light phase at the 1-week recovery time point. Eight weekly phase advances, with 1-week recovery times between each phase advance, further disrupted the H3K4me3 rhythms. Finally, we used the mPer2Luc knockin mouse to determine whether the phase advance also disrupted Per2 protein expression. Similar to the results from the histone work, we found both a rapid response to the phase advance and a delayed recovery, the latter in sync with H3K4me3 levels. A model to explain these results is offered.


Prevalence of anaphylaxis among adults admitted to critical care for severe asthma exacerbation.
In asthmatics, making a diagnosis of anaphylaxis could be challenging as respiratory symptoms are a common feature of both conditions. Identifying anaphylaxis is important to providing appropriate care. A prior study showed that anaphylaxis is sometimes misdiagnosed as acute asthma in children. We sought to identify the percentage of adults admitted to the intensive care unit (ICU) with asthma exacerbations who met criteria for anaphylaxis. Retrospective chart review of adults admitted with acute asthma to the ICU at Jacobi Medical Center, a Level 1 trauma centre in Bronx, New York. Study period was January 2012 to December 2014. Using the criteria outlined in the World Allergy Organization's Anaphylaxis Guidelines, we identified patients who met criteria for anaphylaxis. 105 patients were identified: 17 were excluded because their main reason(s) for admission was not asthma. 7 (8%) of the 88 cases eligible for the study met diagnostic criteria for anaphylaxis while 3 (3.4%) were highly likely to have had anaphylaxis. The baseline characteristics of the seven patients were similar to that of the overall population studied. They however had shorter onset of symptoms (median (IQR): 3.5 (2-6) vs 24 hours (12-72), p<0.001), were more likely to have been intubated (71%vs31%, p=0.04), received intramuscular epinephrine by the emergency medical services or in the emergency room (86%vs42%, p=0.04) and received antihistamines (28.6%vs1.2%, p=0.02). None of these seven patients died. Only one of the seven patients was prescribed an epinephrine pen injector and referred to an allergist at discharge. In this single-centre retrospective review, 3.4% of adults admitted to the ICU for acute severe asthma also met criteria for anaphylaxis.


Preventing catheter-associated urinary tract infections in acute care: the bundle approach.
Catheter-associated urinary tract infections account for 40% of all nosocomial infections. A multidisciplinary team implemented evidence-based guidelines and a urinary catheter bundle, focusing on optimizing the use of urinary catheters through continual assessment and prompt catheter removal. Data were obtained on catheter device days, compliance with urinary catheter orders, and computer documentation of continued catheter indications. Results included an overall reduction of 71% in catheter device days and a 56% reduction in catheter use.


Using workers' compensation claims to investigate occupational injuries among residential day laborers and domestic workers.
While many states have legal provisions to extend workers' compensation eligibility to day laborers or domestic workers hired directly by homeowners or families, little is known about the nature or frequency of injury claims submitted on behalf of these "residential employees." We examined California workers' compensation claims records for injuries that appear to have occurred in residential settings between 2008 and 2018 and where the employer of record was an individual or family. We examined the demographic, occupational, and injury characteristics of our sample and analyzed factors that contribute to the likelihood of a claim resulting in some sort of payment. We identified 5,463 workers' compensation claims that were likely submitted on behalf of residential employees. Claims most commonly reflected injuries to workers performing housekeeping tasks, followed by construction/maintenance and caregiving. Workers performing construction/maintenance or gardening/landscaping tasks were more likely to be injured on the same day of hire and were more likely to require hospitalization or emergency treatment for their injuries; however, these workers also had a smaller proportion of claims that resulted in payment. We found that employment tenure had the biggest impact on the likelihood of payment, with the odds increasing sharply after the first day of work. Although the claims in our data set likely represent only a small fraction of all injuries to residential day laborers and domestic workers during this period, we suggest that workers' compensation claims can provide valuable clues for better understanding occupational injuries among workers in this largely informal sector.


Thin aggregative fimbriae and cellulose enhance long-term survival and persistence of Salmonella.
Salmonella spp. are environmentally persistent pathogens that have served as one of the important models for understanding how bacteria adapt to stressful conditions. However, it remains poorly understood how they survive extreme conditions encountered outside their hosts. Here we show that the rdar morphotype, a multicellular phenotype characterized by fimbria- and cellulose-mediated colony pattern formation, enhances the resistance of Salmonella to desiccation. When colonies were stored on plastic for several months in the absence of exogenous nutrients, survival of wild-type cells was increased compared to mutants deficient in fimbriae and/or cellulose production. Differences between strains were further highlighted upon exposure to sodium hypochlorite, as cellulose-deficient strains were 1,000-fold more susceptible. Measurements of gene expression using luciferase reporters indicated that production of thin aggregative fimbriae (Tafi) may initiate formation of colony surface patterns characteristic of the rdar morphotype. We hypothesize that Tafi play a role in the organization of different components of the extracellular matrix. Conservation of the rdar morphotype among pathogenic S. enterica isolates and the survival advantages that it provides collectively suggest that this phenotype could play a role in the transmission of Salmonella between hosts.


A space-time analysis of the WikiLeaks Afghan War Diary: a resource for analyzing the conflict-health nexus.
Although it is widely acknowledged that areas of conflict are associated with a high health burden, from a geospatial perspective it is difficult to establish these patterns at fine scales because of a lack of data. The release of the "WikiLeaks" Afghan War Diary (AWD) provides an interesting opportunity to advance analysis and theory into this interrelationship. This paper will apply two different space time analyses to identify patterns of improvised explosive devices (IED) detonations for the period of 2004 to 2009 in Afghanistan. There is considerable spatial and temporal heterogeneity in IED explosions, with concentrations often following transportation links. The results are framed in terms of a resource for subsequent analyses to other existing health research in Afghanistan. To facilitate this, in our discussion we present a Google Earth file of overlapping rates that can be distributed to any researcher interested in combining his/her fine scale health data with a similarly granular layer of violence. The release of the AWD presents a previously unavailable opportunity to consider how spatially detailed data about violence can be incorporated into understanding, and predicting, health related spillover effects. The AWD can enrich previous research conducted on Afghanistan, and provide a justification for future "official" data sharing at appropriately fine scales.


Diagnosis of GH deficiency in the transition period: accuracy of insulin tolerance test and insulin-like growth factor-I measurement.
A consensus exists that severe growth hormone deficiency (GHD) in adults is defined by a peak GH response to insulin-induced hypoglycemia (insulin tolerance test, ITT) of less than 3 microg/l based on a cohort of subjects with a mean age of 45 years. By considering one of the following two criteria for the diagnosis of probable permanent GHD, i.e. the severity of GHD (suggested by the presence of multiple pituitary hormone deficiencies (MPHD)) or the magnetic resonance (MR) imaging identification of structural hypothalamic-pituitary abnormalities, 26 patients (17 males, 9 females, mean age 20.8 +/- 2.3 years, range 17-25 years) were selected for re-evaluation of the GH response to ITT and their IGF-I concentration. Eight subjects had isolated GHD (IGHD) and 18 had MPHD. Normative data for peak GH were obtained after ITT in 39 healthy subjects (mean age 21.2 +/- 4.4 years, range 15.1-30.0 years) and the reference range for IGF-I was calculated using normative data from 117 healthy individuals. Mean peak GH response to ITT was significantly lower in the 26 patients (1.8+/-2.0 microg/l, range 0.1-6.1 microg/l) compared with the 39 controls (18.5 +/- 15.5 microg/l, range 6.1-84.0 microg/l; P < 0.0001). One subject with septo-optic dysplasia had a peak GH response of 6.1 microg/l that overlapped the lowest peak GH response obtained in normal subjects. There was an overlap for IGF-I SDS between subjects with IGHD and MPHD, as well as with normal controls. The diagnostic accuracy of a peak GH response of 6.1 microg/l showed a 96% sensitivity with 100% specificity. The maximum diagnostic accuracy with IGF-I SDS was obtained with a cut-off of -1.7 SDS (sensitivity 77%, specificity 100%) while an IGF-I < or = - 2.0 SDS showed a sensitivity of 62%. Our data show that the cut-off value of the peak GH response to ITT of less than 3 microg/l or 5 microg/l and of IGF-I of less than -2.0 SDS are too restrictive for the diagnosis of permanent GH deficiency in the transition period. We suggest that permanent GHD could be investigated more accurately by means of an integrated analysis of clinical history, the presence of MPHD, IGF-I concentration and the MR imaging findings of structural hypothalamic-pituitary abnormalities.


[Acute liver failure in a hospital with a liver transplantation program].
Acute hepatic failure (AHF) is an uncommon entity but with high mortality. Liver transplantation has improved prognosis but is an aggressive treatment with high risk. Currently, there are no accurate criteria to differentiate between irreversible AHF and the possibility of hepatic regeneration. The aim of this study is to review the clinical characteristics indicating transplantation and the outcome of AHF in patients at our institution. Descriptive and retrospective study of cases of AHF at our institution in the last 3 years. Eleven patients were studied. Mortality was lower in transplant recipients (23%; 2 out of 8) than in non-transplanted patients (67%; 2 out 3). Hepatic regeneration occurred in one patient. Overall mortality was 36% with a perioperative mortality of 13%. The indication of hepatic transplantation in patients with a poor prognosis, early stage acute liver failure, and a low grade of encephalopathy, and prior to the development of multiorgan failure could improve the results of transplantation and reduce perioperative mortality.