Purification, characterization and crystallization of a group of earthworm fibrinolytic enzymes from Eisenia fetida.
Seven fibrinolytic enzymes were purified from the earthworm Eisenia fetida. The molecular weights of the enzymes were 24663, 29516, 29690, 24201, 24170, 23028 and 29595, and the respective isoelectric points were 3.46, 3.5, 3.5, 3.68, 3.62, 3.94 and 3.46. All the proteases showed different fibrinolytic activity on fibrin plates. Studies on substrate specificity and inhibition indicated that they belonged to different types of serine proteases. N-Terminal sequencing indicated their high homology to those from the earthworm Lumbricus rubellus. All the enzymes have been crystallized.


Features of Myocardial Remodeling and Changes in the Blood Lipid Spectrum in Experimental Doxorubicin-Induced Cardiomyopathy and Atorvastatin Administration.
Structural myocardial reorganization and changes in the blood lipid spectrum in rats were studied after administration of a single sublethal dose of doxorubicin (15 mg/kg) alone and in combination with atorvastatin (20 mg/kg/day over 7 days). It was established that doxorubicin induced the development of dyslipidemia in experimental animals (the concentrations of total cholesterol, triglycerides, and VLDL increased by 2.2, 2.0, and 1.96 times, respectively; the atherogenic coefficient increased by 3.4 times by day 7 of the experiment). In animals with experimental anthracycline cardiomyopathy treated with atorvastatin, the concentrations of the main components of the blood lipid spectrum increased less markedly. Atorvastatin alone induces moderate myocardial remodeling in comparison with more pronounced changes in the structural organization of the myocardium in rats treated with doxorubicin alone. Course treatment with atorvastatin under conditions of doxorubicin-induced cardiomyopathy reduced the severity of myocardial remodeling: the decrease in the volume density of cardiomyocytes and the increase in the volume density of the connective tissue were less pronounced in the dynamics of the experiment.


In vivo analysis of voltage-dependent calcium channels.
The molecular cloning of calcium channel subunits has identified an unexpectedly large number of genes and splicing variants, many of whichhave complex expression patterns: a central problem of calcium channel biology is to understand the functional significance of this genetic complexity. The genetic analysis of voltage-dependent calcium channels (VDCCs) provides an approach to defining channel function that is complimentary to pharmacological, electrophysiological, and other molecular methods. By discovering or creating alleles of VDCC genes, one can gain an understanding of the VDCC function at the whole animal level. Of particular interest are mutations in the alpha1 genes that encode the pore forming subunits, as they define the specific channel subtypes. In fact, a variety of calcium channelopathies and targeted mutations have been described for these genes in the last 6 years. The mutant alleles described below illustrate how phenotype analysis of these alleles has uncovered very specific functional roles that can be localized to specific synapses or cells.


IL-1beta and IL-6 modulate apolipoprotein E gene expression in rat hepatocyte primary culture.
Incubation of rat hepatocytes in primary culture with IL-1beta at a concentration of 2.5 units/ml resulted in an increase (+80%) in the amount of apoE mRNA without any effect upon apoE synthesis. IL-6 at a low concentration (10 units/ml) induced a decrease (-35%) in the amount of apoE mRNA, but increased apoE synthesis (+28%). No effect was observed with higher concentrations of IL-1beta (10 units/ml) or IL-6 (100 units/ml). These results suggest that inflammatory cytokines IL-1beta and IL-6 modulate the expression of apoE gene in cultured rat hepatocytes, at a concentration that does not induce the acute phase response.


A history of smoking does not reduce long-term benefits of cardiac rehabilitation on cardiorespiratory fitness in men and women with cardiovascular disease.
Smoking is an important risk factor for cardiovascular disease and all-cause mortality. Cardiac rehabilitation (CR) is effective for reducing the risk of recurrent cardiac events through improving cardiorespiratory fitness (CRF). Little is known about the influence of smoking on CRF throughout long-term CR. The purpose of this analysis was to compare CRF trajectories among individuals with positive and negative smoking history enrolled in long-term CR. Participants had a positive smoking history if they currently or formerly smoked (Smoke+, n = 55, mean age = 64.9 ± 9.0 years) and had a negative history if they never smoked (Smoke-, n = 34, mean age = 61.4 ± 9.0 years). CRF (peak oxygen uptake) was measured at baseline and annually thereafter for 6 years. The Smoke+ group had lower CRF compared with the Smoke- group over enrollment (β = -3.29 (SE = 1.40), 95% confidence interval (CI) -6.04 to -0.54, p = 0.02), but there was no interaction of smoking history and enrollment (β = 0.35 (SE = 0.21), 95% CI: -0.06 to 0.77, p = 0.10). Moreover, trajectories were not influenced by pack-years (β = 0.01 (SE = 0.01), 95% CI: -0.01 to 0.04, p = 0.23) or time smoke-free (β = -0.002 (SE = 0.01), 95% CI: -0.02 to 0.02, p = 0.80). Although the trajectories of CRF do not appear to be affected by smoking behaviour, individuals without a history of smoking maintained higher CRF throughout enrollment. Novelty: The benefits of long-term exercise-based cardiac rehabilitation on cardiorespiratory fitness are similar between those who have smoked and those who have never smoked. Neither the number of pack-years nor the length of time spent smoke-free influence cardiorespiratory fitness trajectories following long-term cardiac rehabilitation.


The Cockayne-Touraine type of dominant dystrophic epidermolysis bullosa--ultrastructural similarities to the Pasini variant.
Biopsies from the uninvolved skin of 5 patients with the Cockayne-Touraine (CT) type of dystrophic epidermolysis bullosa (DEB) were studied with the electron microscope. Dermal fibrillar bodies were noted in 2 patients and 3 showed basal lamina (BL) duplication or splitting. Discontinuity of the BL with herniation of keratinocyte cytoplasm was present in one patient. These changes, thought previously to be characteristic of the Pasini variant, indicate that abnormalities of the BL may be involved in blister formation in both disease subgroups.


Intracranial lesion features in moderate-to-severe traumatic brain injury: relation to neurointensive care variables and clinical outcome.
The primary aim was to determine the association of intracranial hemorrhage lesion type, size, mass effect, and evolution with the clinical course during neurointensive care and long-term outcome after traumatic brain injury (TBI). In this observational, retrospective study, 385 TBI patients treated at the neurointensive care unit at Uppsala University Hospital, Sweden, were included. The lesion type, size, mass effect, and evolution (progression on the follow-up CT) were assessed and analyzed in relation to the percentage of secondary insults with intracranial pressure > 20 mmHg, cerebral perfusion pressure < 60 mmHg, and cerebral pressure autoregulatory status (PRx) and in relation to Glasgow Outcome Scale-Extended. A larger epidural hematoma (p < 0.05) and acute subdural hematoma (p < 0.001) volume, greater midline shift (p < 0.001), and compressed basal cisterns (p < 0.001) correlated with craniotomy surgery. In multiple regressions, presence of traumatic subarachnoid hemorrhage (p < 0.001) and intracranial hemorrhage progression on the follow-up CT (p < 0.01) were associated with more intracranial pressure-insults above 20 mmHg. In similar regressions, obliterated basal cisterns (p < 0.001) were independently associated with higher PRx. In a multiple regression, greater acute subdural hematoma (p < 0.05) and contusion (p < 0.05) volume, presence of traumatic subarachnoid hemorrhage (p < 0.01), and obliterated basal cisterns (p < 0.01) were independently associated with a lower rate of favorable outcome. The intracranial lesion type, size, mass effect, and evolution were associated with the clinical course, cerebral pathophysiology, and outcome following TBI. Future efforts should integrate such granular data into more sophisticated machine learning models to aid the clinician to better anticipate emerging secondary insults and to predict clinical outcome.


Identification of a novel alternative splicing variant of hemocyanin from shrimp Litopenaeus vannamei.
Recent evidences suggest that invertebrates express families of immune molecules with high levels of sequence diversity. Hemocyanin is an important non-specific immune molecule present in the hemolymph of both mollusks and arthropods. In the present study, we characterized a novel alternative splicing variant of hemocyanin (cHE1) from Litopenaeus vannamei that produced mRNA transcript of 2579 bp in length. The isoform contained two additional sequences of 296 and 267 bp in the 5'- and 3'-terminus respectively, in comparison to that of wild type hemocyanin (cHE). Sequence of cHE1 shows 100% identity to that of hemocyanin genomic DNA (HE, which does not form an open reading frame), suggesting that cHE1 might be an alternative splicing variant due to intron retention. Moreover, cHE1 could be detected by RT-PCR from five tissues (heart, gill, stomach, intestine and brain), and from shrimps at stages from nauplius to mysis larva. Further, cHE1 mRNA transcripts were significantly increased in hearts after 12h of infection with Vibrio parahemolyticus or poly I: C, while no significant difference in the transcript levels of hepatopancreas cHE was detected in the pathogen-treated shrimps during the period. In summary, these studies suggested a novel splicing variant of hemocyanin in shrimp, which might be involved in shrimp resistance to pathogenic infection.


Yeast haspin kinase regulates polarity cues necessary for mitotic spindle positioning and is required to tolerate mitotic arrest.
Haspin is an atypical protein kinase that in several organisms phosphorylates histone H3Thr3 and is involved in chromosome segregation. In Saccharomyces cerevisiae, H3Thr3 phosphorylation has never been observed and the function of haspin is unknown. We show that deletion of ALK1 and ALK2 haspin paralogs causes the mislocalization of polarisome components. Following a transient mitotic arrest, this leads to an overly polarized actin distribution in the bud where the mitotic spindle is pulled. Here it elongates, generating anucleated mothers and binucleated daughters. Reducing the intensity of the bud-directed pulling forces partially restores proper cell division. We propose that haspin controls the localization of polarity cues to preserve the coordination between polarization and the cell cycle and to tolerate transient mitotic arrests. The evolutionary conservation of haspin and of the polarization mechanisms suggests that this function of haspin is likely shared with other eukaryotes, in which haspin may regulate asymmetric cell division.


Development and Innovation of Ganoderma Industry and Products in China.
Ganoderma (Lingzhi) has been used as a medicinal mushroom to promote health in China for more than 2000 years. The modern research and development of Ganoderma industry started from the 1950s, in which the successful cultivation of Ganoderma fruiting body and submerged fermentation of Ganoderma mycelium lay the critical foundation for the industry development. Recent decades have witnessed the rapid development of Ganoderma industry, which is boosted through various efforts made by the government, the academia, and the industry. In this chapter, the development of Ganoderma industry in China is reviewed in terms of gross output, standards, scientific articles, patents, and associations. In addition, development of Ganoderma products and manufacturing technologies are also overviewed and summarized. In the last section, several innovation trends are suggested for the further development of Ganoderma industry.


Increased stromal extracellular matrix synthesis and assembly by insulin activated bovine keratocytes cultured under agarose.
Previously, pharmacological levels of insulin have been shown to stimulate the synthesis of normal corneal stromal collagen and proteoglycans by bovine keratocytes in culture. Here we compared insulin to physiological levels of IGF-I and found that IGF-I also stimulated the synthesis of these extracellular matrix components, but less than that of insulin. Keratocytes in monolayer culture secreted most of the collagen synthesized into the media in the form of procollagen, a precursor of collagen. We found that an overlay of 3% agarose on the keratocytes in culture enhanced the conversion of procollagen to collagen and increased the deposition of collagen and proteoglycans into the cell layer. The extracellular matrix associated with the keratocytes cultured under agarose exhibited a corneal stromal-like architecture. These results suggest that enhancing the conversion of procollagen to collagen is a key step in the formation of extracellular matrix by keratocytes in vitro. Agarose overlay of insulin activated keratocytes in culture is a useful model for studying corneal stromal extracellular matrix assembly in vitro.


Transfusion therapy for autoimmune hemolytic anemia patients: a laboratory perspective.
Patients presenting with autoimmune hemolytic anemias create inherent challenges to those tasked with providing compatible blood for transfusion therapy. These patients have developed autoantibodies against their own red cell surface antigens. Because these antigens are usually high-incidence, these patients will typically demonstrate panagglutination when their serum is exposed to most commercially procured screening red blood cells. This makes the identification of clinically significant alloantibodies difficult for laboratory personnel. Transfusion history, patient phenotype availability, and previous antibody records all impact the testing methods. The end goal is to identify clinically significant alloantibodies in order to provide antigen negative, compatible red blood cells, which reduces the risk of transfusion related reactions. It is imperative to understand the laboratory results and the techniques available that guide the investigative process.


Taxonomic status of purported primate frontal bones from the Eocene Pondaung Formation of Myanmar.
Two isolated cranial fragments from the late middle Eocene Pondaung Formation of central Myanmar have previously been interpreted as frontal bones of the amphipithecid primate Amphipithecus mogaungensis. Aside from a few maxillary fragments, these specimens provide the only potential source of information currently available regarding the cranial anatomy of Amphipithecidae. Were this taxonomic attribution correct, these specimens would indicate that amphipithecids retained numerous primitive skull features, including the absence of a postorbital septum, the retention of a voluminous olfactory chamber, and strong separation between the forebrain and the orbital fossa. However, several anatomical details observable on these specimens are incompatible with their attribution to any primate and strongly suggest that they cannot be ascribed to Mammalia. Particularly problematic in this regard are the extreme thickness of the dermal bone, the odd structure of the alleged "frontal trigon," and the mediolateral orientation and uniquely robust construction of the descending process of the frontal bone (which partially segregates the orbital and temporal fossae). Because these isolated elements can no longer be attributed to Amphipithecus, the anatomical, phylogenetic, and behavioral inferences regarding amphipithecid paleobiology that have been drawn from these specimens can no longer be sustained.


A new correction method for determination on carbohydrates in lignocellulosic biomass.
The accurate determination on the key components in lignocellulosic biomass is the premise of pretreatment and bioconversion. Currently, the widely used 72% H2SO4 two-step hydrolysis quantitative saccharification (QS) procedure uses loss coefficient of monosaccharide standards to correct monosaccharide loss in the secondary hydrolysis (SH) of QS and may result in excessive correction. By studying the quantitative relationships of glucose and xylose losses during special hydrolysis conditions and the HMF and furfural productions, a simple correction on the monosaccharide loss from both PH and SH was established by using HMF and furfural as the calibrators. This method was used to the component determination on corn stover, Miscanthus and cotton stalk (raw materials and pretreated) and compared to the NREL method. It has been proved that this method can avoid excessive correction on the samples with high-carbohydrate contents.


In vitro selection of phosphorothiolated aptamers.
A pool of RNA molecules that contained exclusively phosphorothioate internucleoside linkages was used as a starting point for the selection of aptamers that bind to basic fibroblast growth factor (bFGF), and appear to act as heparin mimics.


Neurophysiological signatures of sensory-processing sensitivity.
Sensory processing sensitivity is mainly captured based on questionnaires and it's neurophysiological basis is largely unknown. As hitherto no electroencephalography (EEG) study has been carried out, the aim of this work was to determine whether the self-reported level of SPS correlates with the EEG activity in different frequency bands. One hundred fifteen participants were measured with 64-channel EEG during a task-free resting state. After artifact correction, a power spectrum time series was calculated using the Fast Fourier Transform (FFT) for the following frequency bands: Delta: 1-3.5 Hz, theta: 4-7.5 Hz, alpha1: 8-10 Hz, alpha2: 10.5-12 Hz, beta1: 12.5-15 Hz, beta2: 15.5-25 Hz, gamma: 25.5-45 Hz, global: 1-45 Hz. Correlations with the 'Highly Sensitive Person Scale' (HSPS-G) scores were determined. Then, the lowest and the highest 30% of the cohort were contrasted as polar opposites. EEG features were compared between the two groups applying a paired two-tailed t-test. The HSPS-G scores correlated statistically significantly positive with beta 1 and 2, and global EEG power during resting with eyes open, but not during resting with eyes closed. The highly sensitive group revealed higher beta power (4.38 ± 0.32 vs. 4.21 ± 0.17, p = 0.014), higher gamma power (4.21 ± 0.37 vs. 4.00 ± 0.25, p = 0.010), and increased global EEG power (4.38 ± 0.29 vs. 4.25 ± 0.17, p = 0.041). The higher EEG activity in the HSP group was most pronounced in the central, parietal, and temporal region, whereas lower EEG activity was most present in occipital areas. For the first time, neurophysiological signatures associated with SPS during a task free resting state were demonstrated. Evidence is provided that neural processes differ between HSP and non-HSP. During resting with eyes open HSP exhibit higher EEG activity suggesting increased information processing. The findings could be of importance for the development of biomarkers for clinical diagnostics and intervention efficacy evaluation.


Impact of chemical exposure on the fish Pomatoschistus microps Krøyer (1838) in estuaries of the Portuguese Northwest coast.
Juveniles of the estuarine fish Pomatoschistus microps were collected from autumn 2001 to summer 2002 in five stations along the Portuguese Northwest coast with different types and/or levels of environmental contamination: two reference sites with low levels of contamination (R1 and R2) and three differently impacted areas with higher levels of contamination. UI is located in an estuary under the influence of urban and industrial effluents, AA in a channel that receives intensive agriculture run-off and IE in a highly impacted industrial area. The activity of the enzymes acetylcholinesterase (AChE), lactate dehydrogenase (LDH), 7-ethoxyresorufin O-deethylase (EROD) and glutathione S-transferases (GST) were used as environmental biomarkers on P. microps. A significant seasonality effect on all the enzymatic activities was found, lower levels being registered in winter and spring on AChE, in autumn on LDH, and in winter on GST and EROD. The battery of biomarkers used was capable of discriminating sites with different types and/or levels of contamination, R1 and UI being the highest discriminated (91.7% and 66.7%, respectively). At R1 significantly lower levels of AChE and LDH were found, and EROD was significantly inducted at UI. Furthermore, IE presented higher levels of GST, and R2 and AA an inhibition of AChE in winter and spring. The results indicated that the battery of biomarkers used in this study seems to be a useful tool to distinguish between different types of environmental contamination in estuarine systems, and that P. microps is a suitable species to be used as bioindicator.


Fresh water resource, scarcity, water salinity challenges and possible remedies: A review.
Water is one of the natural resource due balance if our planet and the life on it have to sustain and economic development to be expected in the future. The increase in population of the world and level of wealth of humans is expected to withdraw more freshwater. However, since water is already one of the limited resources, global per capital water available surely drops and water shortage happens. Pollution of ground and surface water by dissolved salts are increasing and exacerbating this water shortage situation. The sources of these dissolved salts (such as primary and secondary salinity-causing agents) are known to change the chemical constituent of water. Once contributing factors for water scarcity are identified, future man should work on it to overcome the challenge. This paper therefore began with global water resource information and indicated different levels of scarcity to give overall clues on the situation. Salinity description, its global status, causative factors and challenges were revised before possible recommendations were indicated as indispensable solution.


Effectiveness of amiodarone on ventricular arrhythmias during and after acute myocardial infarction.
A randomized, placebo-controlled study examined the effect of amiodarone on the incidence of ventricular arrhythmias after acute myocardial infarction (AMI). Two hundred patients with AMI were randomized to receive amiodarone, 200 mg every 8 hours for 1 month, followed by 200 mg/day, or placebo, starting 48 hours after the onset of chest pain. Twenty-four-hour Holter monitoring was performed on day 1, repeated after 8 days and subsequently at 3-month intervals. One hundred seventy-two patients were followed for 6 to 42 months and monitor data were available at 6 to 9 months in 129 patients. Amiodarone was well tolerated, with no serious side effects; 12 patients were withdrawn from therapy. At 6 to 9 months more than 1 ventricular premature complex per hour was present in 3 of the 59 amiodarone-treated patients (5%) and 24 of the 70 placebo-treated patients (34%) (p less than 0.02). Complex arrhythmias (multifocal, early cycle, repetitive, bigeminy, trigeminy or ventricular tachycardia) were present in 5 of 59 of the amiodarone-treated patients (8%) and 20 of 70 placebo-treated patients (28%) (p less than 0.005). Sixteen patients in the amiodarone group and 11 in the placebo group died (difference not significant). Thus, amiodarone can reduce the frequency and complexity of ventricular arrhythmias after AMI, but it is unlikely that this will result in a substantial beneficial effect on post-AMI mortality risk.


Budd-Chiari syndrome mimicking autoimmune hepatitis.
Budd-Chiari syndrome is a rare disorder characterized by obstruction of hepatic venous outflow involving either the hepatic veins or the inferior vena cava. Risk factors include thrombophilia and myeloproliferative disorders, and symptoms are often unspecific. We present a case of a 60-year-old woman admitted to our medical unit for ascites and abdominal pain. Although her medical history significant for a mixed connective tissue disease and a mild increase of the transaminases had raised the suspicion of autoimmune hepatitis, the demonstration on computed tomography of the absence of outflow in the terminal tract of the supra-hepatic veins allowed a correct diagnosis. The contribution of radiological imaging is fundamental in the diagnosis of this rare and elusive pathology.


Relative importance of different human aPL derived heavy and light chains in the binding of aPL to cardiolipin.
Previous studies have suggested the importance of somatic mutations and arginine, asparagine and lysine residues in the complementarity determining regions (CDRs) of antiphospholipid antibodies (aPL) implicated in the pathogenesis of the antiphospholipid antibody syndrome. The relative contributions of the heavy and light chains of aPL in binding to cardiolipin (CL) were assessed by pairing the heavy and light chains of two IgG, beta(2)GPI dependent aPL (IS4 and CL24) with different partner chains from other IgG, beta(2)GPI independent aPL (UK4) and anti-DNA antibodies (B3 and 33H11). Four heavy (V(H)) and five light (V(L)) chain variable sequences from three aPL and two anti-DNA antibodies were cloned into expression vectors containing appropriate gamma(1), lambda or kappa constant region cDNA. Paired combinations of heavy and light chain expression plasmids were transfected into COS-7 cells allowing transient expression of whole IgG molecules, which were harvested and tested for the ability to bind CL and DNA by enzyme-linked immunosorbant assay (ELISA). Whole IgG was produced from 19 heavy/light chain combinations. IS4V(H) was dominant in conferring the ability to bind CL with four of the five V(L) tested. The identity of the V(L) region paired with IS4V(H) was important in determining the strength of binding to CL. IS4V(H) contains multiple arginine residues in CDR3, which may have accumulated due to antigen driven selection. It is likely that these arginine residues may interact with CL. The combination B3V(H)/B3V(L) also bound CL, but none of the other 14 combinations showed any binding in this assay. Whole IgG molecules capable of binding CL were produced by in vitro expression in COS-7 cells. Arginine residues play important roles in binding to CL and double-stranded DNA. However, different patterns of mutation to arginine are associated with binding to each of these antigens.


Optical Coherence Tomography-Guided or Intravascular Ultrasound-Guided Percutaneous Coronary Intervention: The OCTIVUS Randomized Clinical Trial.
Intravascular imaging-guided percutaneous coronary intervention (PCI) with intravascular ultrasound (IVUS) or optical coherence tomography (OCT) showed superior clinical outcomes compared with angiography-guided PCI. However, the comparative effectiveness of OCT-guided and IVUS-guided PCI regarding clinical outcomes is unknown. In this prospective, multicenter, open-label, pragmatic trial, we randomly assigned 2008 patients with significant coronary artery lesions undergoing PCI in a 1:1 ratio to undergo either an OCT-guided or IVUS-guided PCI. The primary end point was a composite of death from cardiac causes, target vessel-related myocardial infarction, or ischemia-driven target-vessel revascularization at 1 year, which was powered for noninferiority of the OCT group compared with the IVUS group. Safety outcomes were also assessed. At 1 year, primary end point events occurred in 25 of 1005 patients (Kaplan-Meier estimate, 2.5%) in the OCT group and in 31 of 1003 patients (Kaplan-Meier estimate, 3.1%) in the IVUS group (absolute difference, -0.6 percentage points; upper boundary of one-sided 97.5% CI, 0.97 percentage points; P<0.001 for noninferiority). The incidence of contrast-induced nephropathy was similar (14 patients [1.4%] in the OCT group versus 15 patients [1.5%] in the IVUS group; P=0.85). The incidence of major procedural complications was lower in the OCT group than in the IVUS group (22 [2.2%] versus 37 [3.7%]; P=0.047), although imaging procedure-related complications were not observed. In patients with significant coronary artery lesions, OCT-guided PCI was noninferior to IVUS-guided PCI with respect to the incidence of a composite of death from cardiac causes, target vessel-related myocardial infarction, or ischemia-driven target-vessel revascularization at 1 year. The selected study population and lower-than-expected event rates should be considered in interpreting the trial. URL: https://www. gov; Unique number: NCT03394079.


Do arginine metabolites have a role in periodontitis due to smoking? A new perspective.
Cigarette consumption is common around the world and besides its negative effects on health, and its effects on periodontitis draw attention. Arginine metabolites are involved in the pathogenesis of several systemic inflammatory diseases' including cardiovascular diseases. Our aim was to determine periodontitis and healthy individuals' arginine metabolites and IL-6 levels in saliva and serum and to evaluate those according to smoking status. The study consisted of four groups: healthy individuals (control [C]; n = 20), smokers with healthy periodontium (S-C; n = 20), nonsmokers with Stage-III Grade-B generalized periodontitis (P; n = 20) and smokers with Stage-III Grade-C generalized periodontitis (S-P; n = 18). Periodontal parameters were measured. Analysis of methylated arginine metabolites was performed by LC-MS/MS, and IL-6 levels were determined by ELISA kits. In nonsmokers, salivary concentrations of asymmetric dimethylarginine (ADMA) and symmetrical dimethylarginine (SDMA) were higher in the periodontitis than control (p < 0.001, p = 0.010). Smokers with periodontitis exhibited higher ADMA (p = 0.033, p < 0.001) and arginine (p = 0.030, p = 0.001) saliva concentrations than smoking and nonsmoking controls. Our results demonstrated that salivary concentrations of ADMA and SDMA were associated with periodontitis. Smoking increased ADMA, SDMA and NG -monomethyl L-arginine (L-NMMA) levels in serum only in periodontitis patients.


Revisiting Competing Paths in Electrochemical CO2 Reduction on Copper via Embedded Correlated Wavefunction Theory.
We re-evaluate two key steps in the mechanism of CO2 reduction on copper at a higher level of theory capable of correcting inherent errors in density functional theory (DFT) approximations, namely, embedded correlated wavefunction (ECW) theory. Here, we consider the CO reduction step on Cu(111), which is critical to understanding reaction selectivity. We optimize embedding potentials at the periodic plane-wave DFT level using density functional embedding theory (DFET). All possible adsorption sites (adsites) for each adsorbate then are screened with ECW theory at the catalytically active site to refine the local electronic structure. Unsurprisingly, DFT and ECW theory predict different adsite preferences, largely because of DFT's inability to properly situate the CO 2π* level. Differing preferred adsites suggest that different reaction pathways could emerge from DFT versus ECW theory. Starting from these preferred ECW theory adsites, we then obtain reaction pathways at the plane-wave DFT level using the climbing-image nudged elastic band method to determine minimum energy paths. Thereafter, we perform ECW calculations at the catalytically active site to correct the energetics at each interpolated structure (image) along the reaction pathways. Via this approach, we confirm that the first step in CO reduction via hydrogen transfer on Cu(111) is to form hydroxymethylidyne (*COH) instead of formyl (*CHO). Although the prediction to preferentially form *COH is consistent with that of DFT, the two theories predict quite different structural and mechanistic behaviors, suggesting that verification is needed for other parts of the mechanism of CO2 reduction, which is the subject of ongoing work.


Reciprocal regulation of the Ca2+ and H+ sensitivity in the SLO1 BK channel conferred by the RCK1 domain.
Increasing evidence suggests that intracellular H+ directly stimulates large-conductance Ca2+- and voltage-activated K+ (SLO1 BK) channels, thus providing a crucial link between membrane excitability and cell metabolism. Here we report that two histidine residues, His365 and His394, located in the intracellular regulator of conductance for K+ (RCK) 1 domain, serve as the H+ sensors of the SLO1 BK channel. Activation of the channel by H+ requires electrostatic interactions between the histidine residues and a nearby negatively charged residue involved in the channel's high-affinity Ca2+ sensitivity. Reciprocally, His365 and His394 also participate in the Ca2+-dependent activation of the channel, functioning as Ca2+ mimetics once they are protonated. Therefore, a common motif in the RCK1 domain mediates the stimulatory effects of both H+ and Ca2+, and provides a basis for the bidirectional coupling of cell metabolism and membrane electrical excitability.


Effects of cromakalim on acetylcholine release and smooth muscle contraction in guinea-pig small intestine.
The effects of the potassium channel opener cromakalim on smooth muscle contraction and 3H-acetyl-choline release were studied simultaneously in guinea-pig longitudinal muscle myenteric plexus preparations which had been preincubated with 3H-choline. Cromakalim (10 mumol/l) inhibited more markedly the smooth muscle contractions caused by the release of endogenous acetylcholine (via electrical stimulation or via activation of nicotine- and 5-HT3-receptors) than contractions induced by pilocarpine. Cromakalim (10 mumol/l) did not affect the release of 3H-acetylcholine evoked by electrical stimulation or by stimulation of nicotine- and 5-HT3-receptors. In contrast, the release of 3H-acetylcholine caused by stimulation of M1-receptors was concentration-dependently reduced by cromakalim (1-100 mumol/l). The results suggest that the relaxant effect of cromakalim on smooth muscle contraction is not caused by a reduction of acetylcholine release from myenteric neurones. An opening of cromakalim-sensitive potassium channels may be involved in the inhibition of the M1-receptor mediated acetylcholine release.


Safety and efficacy of liraglutide in patients with type 2 diabetes and end-stage renal disease: protocol for an investigator-initiated prospective, randomised, placebo-controlled, double-blinded, parallel intervention study.
Diabetes is the leading cause of end-stage renal disease (ESRD). Owing to renal clearance, several antidiabetic agents cannot be used in patients with ESRD. The present protocol describes an investigator-initiated trial aiming to test safety and efficacy of treatment with the glucagon-like peptide-1 receptor agonist liraglutide in patients with type 2 diabetes and dialysis-dependent ESRD. Twenty patients with type 2 diabetes and ESRD will be compared with 20 matched patients with type 2 diabetes and normal kidney function in a randomised, parallel, placebo-controlled (1 : 1), double-blinded setting. All participants will receive 12 weeks of daily treatment with liraglutide/placebo in an individually titrated dose of 0.6, 1.2 or 1.8 mg. Over nine visits, plasma liraglutide, glycaemic control, β-cell response, cardiovascular parameters, various biomarkers and adverse events will be assessed. The primary endpoint will be evaluated from dose-corrected plasma trough liraglutide concentration at the final trial visit to determine potential accumulation in the ESRD group. The study has been approved by the Danish Medicines Agency, the Scientific-Ethical Committee of the Capital Region of Denmark and the Danish Data Protection Agency. An external monitoring committee (The Good Clinical Practice Unit at Copenhagen University Hospitals) will oversee the study. The results of the study will be presented at national and international scientific meetings, and publications will be submitted to peer-reviewed journals. ClinicalTrials.gov Identifier: NCT01394341.


Diagnosis and differential diagnosis flow diagram of Chinese post-stroke aphasia types and treatment of post-stroke aphasia.
This review aimed to explore the concept, etiology, classification, classical cortical mapping, assessment, diagnosis and differential diagnosis, treatment, rehabilitation, mechanism, recovery, prognosis, and influencing factors for Chinese post-stroke aphasia (PSA). The review emphasized the necessity and significance of neuroimaging assessment of brain and blood vessels and neuropsychological assessment in diagnosis and differential diagnosis of Chinese PSA. In addition, it suggested and recommended to use "dichotomies of internal and external, and anterior and posterior" as a starting point, based on the anatomic location of brain and blood vessels and their relationship with language area and language disorder. As a result, the formulated Chinese PSA classification was more suitable to guide the clinical treatment of cerebral stroke. Diagnosis, classification, and differential diagnosis of Chinese PSA types were performed according to the "dichotomy" and "four elements." The formulated "flow diagram" enabled to determine the classification of Chinese PSA types. It was beneficial for patients to establish targeted and individualized rehabilitation training plans. This review introduced the use of memantine, piracetam, donepezil, etc. in PSA treatment, evaluated clinical studies conducted in China and abroad, investigated the mechanism of action related to the use of drugs in PSA treatment, and explored the therapeutic mechanism of rehabilitation training. It suggested the use drugs of memantine, piracetam, donepezil, etc. combine with non-pharmacotherapy and rehabilitation training in clinical studies on PSA treatment and also in practical settings.


Evidence-based practice: an idea whose time has come.
This paper defines evidence-based practice, gives a brief history of EBP, addresses nurses' current usage of research, briefly examines the need for evidence-based practice, then addresses what evidence-based practice will mean to the profession. It is concluded that at present Australian nursing is in a stage of pre-evidence-based practice in which most Australian nurses neither read research nor apply research findings to practice. However, a demand for evidence-based practice will likely encourage nurses to use research in practice. Finally, professional strategies will be developed to encourage the introduction and evaluation of evidence-based nursing practice.


Robotic-assisted colectomy in children: a comparative study with laparoscopic surgery.
The aim of this study was to compare outcomes of laparoscopic and robotic-assisted colectomy in children. All children who underwent a colectomy with a laparoscopic (LapC) or robotic-assisted (RobC) approach in our institution (January 2010-March 2023) were included. Demographics, surgical data, and post-operative outcomes within 30 days were collected. Additional cost related to the robotic approach was calculated. Comparisons were performed using Fisher tests for categorical variables and Mann-Whitney tests for continuous variables. A total of 55 colectomies were performed: 31 LapC and 24 RobC (median age: 14.9 years). Main indications included: inflammatory bowel disease (n = 36, 65%), familial adenomatous polyposis (n = 6, 11%), sigmoid volvulus (n = 5, 9%), chronic intestinal pseudo-obstruction (n = 3, 5%). LapC included 22 right, 4 left, and 5 total colectomies. RobC included 15 right, 4 left, and 5 total colectomies. Robotic-assisted surgery was associated with increased operative time (3 h vs 2.5 h, p = 0.02), with a median increase in operative time of 36 min. There were no conversions. Post-operative complications occurred in 35% of LapC and 38% of RobC (p = 0.99). Complications requiring treatment under general anesthesia (Clavien-Dindo 3) occurred in similar rates (23% in LapC vs 13% in RobC, p = 0.49). Length of hospitalization was 10 days in LapC and 8.5 days in RobC (p = 0.39). The robotic approach was associated with a median additional cost of 2156€ per surgery. Robotic-assisted colectomy is as safe and feasible as laparoscopic colectomy in children, with similar complication rates but increased operative times and cost.


A comparison between the effects of nystatin, clotrimazole and miconazole on vaginal candidiasis.
Nystatin (100 000 units/day for 14 days) and clotrimazole (100 mg/day for 6 days) were equivalent to miconazole (100 mg/day for 14 days or 100 mg twice daily for 7 days) in curing vaginal candidiasis. However significantly more patients relapsed in the nystatin and clotrimazole groups than in either of the miconazole groups.


Chunks of charge: effects at play in the assembly of macromolecules at fluid surfaces.
Large macromolecules with hydrophobic backbones are known to assemble at the interface between immiscible liquids. This assembly is often unpredictable because of the subtle interplay among hydrophobic interactions, hydrophilic solvation, structural constraints, and the thermodynamics of adsorption. In these studies, we employ vibrational sum frequency spectroscopy and interfacial tension measurements to study the assembly of a simple polyelectrolyte, poly(methacrylic acid), as it assembles at the interface between two immiscible liquids, specifically, carbon tetrachloride and water. By adjusting the polyelectrolyte charge through pH studies and the polymer size through molecular weight studies, we demonstrate that charge accumulation in segments of the polymer chains is a critical factor in macromolecular interfacial adsorption and desorption. The results have implications for related charged macromolecules whose ability to assemble between two immiscible fluid media is essential for many biological processes, water remediation efforts, and enhanced oil recovery.


Effect of culture conditions and method of conception on mouse live birth rate.
To understand in a mouse model whether there are differences in the decidua and live birth rate after transfer of blastocysts generated by in vitro fertilization (IVF) or by superovulation with spontaneous mating into unstimulated recipients. Animal experiment. University-affiliated tertiary hospital. Mice. IVF embryos were generated and cultured in either Whitten medium (WM, suboptimal conditions) and 20% O2 or KSOM medium with amino acids (KAA, optimal conditions) and 5% O2. The control blastocysts from superovulated mice were flushed out of the uterus 3.5 days (E3.5) after mating (FB group). The resulting blastocysts were transferred to nonsuperovulated CF1 recipients mated to vasectomized males. To understand whether anomalies of decidua were present, the expression of genes involved in decidual development and inflammation was analyzed at E7.5 and E18.5. Similarly, immunostaining was used to evaluate whether the pathways involved in activation of mTORC1 (p-S6) and Cox2 signaling (Cox 2 staining) were altered. Live birth rate, gene expression, and immunostaining of decidua. Implantation rates at E7.5 were similar, but in vivo embryos (FB groups) were predicted to result in live births 3.3 times higher (2.2-5.1) and 6.6 times higher (4.7-9.3) compared with optimal and suboptimal cultures, respectively. Expression of genes involved in decidual development and inflammation or localization and intensity of staining for p-S6 (mTOR pathway), or inflammation (Cox 2 pathway) were not different among the groups. The predicted live birth rate was decreased in mouse embryos generated by IVF compared with embryos generated by mating, whereas the implantation rate was not different. Suboptimal culture conditions resulted in lower birth rate. We did not find evidence of abnormalities in decidualization that could explain these findings. These data indicate that blastocysts cultured in stressful conditions are less competent, suggesting that decreasing the number of embryonic manipulations may result in higher live birth rates.


Stability evaluation of gardenia yellow pigment in presence of different phenolic compounds.
Gardenia yellow pigment (GYP) may undergo chemical degradation under different conditions resulting in color fading. This study investigated the effects of different phenolic compounds (caffeic acid, rosmarinic acid, tannic acid, epicatechin, chlorogenic acid, epigallocatechin, and epigallocatechin gallate) on the physical and chemical stability of GYP under light and different temperatures. Furthermore, food models with GYP/phenolic compounds were simulated to evaluate the GYP stability under different cooking methods. The addition of phenolic compounds, especially tannic acid, epigallocatechin gallate, epigallocatechin, and rosmarinic acid, significantly improved the GYP stability during light and thermal treatments. Fourier transform infrared (FTIR) and nuclear magnetic resonance (NMR) spectroscopy confirmed that the formation of hydrogen bonds between GYP and selected phenolic compounds (tannic acid, epigallocatechin gallate, epigallocatechin, and rosmarinic acid), which may lead to the enhancement of GYP stability. Moreover, these selected phenolic compounds provided potent protective effects on GYP under different cooking methods.


Spontaneous tension hemothorax in a severe COVID-19 patient receiving ECMO therapy: The other side of COVID-19-associated coagulopathy.
As opposed to widely recognized Coronavirus Disease 2019 (COVID-19)-associated thrombotic events, the unusual but serious bleeding complications in COVID-19 patients are worth-mentioned. Here, we describe a 44-year-old man afflicted by COVID-19 pneumonia with acute respiratory distress syndrome (ARDS) and submassive pulmonary embolism. The patient's condition initially improved with the prescription of ECMO, tocilizumab, and hemoadsorption, however, he later developed spontaneous tension hemothorax, which is considered rare but devastating in the setting of COVID-19. While the exact pathogenesis of COVID-19-associated bleeding events remains poorly understood, we aim to highlight the other aspect of coagulation dysfunction potentially caused by COVID-19.


Anger vs guilt: inference of responsibility attribution and interpersonal reactions.
This study examined reactions toward an expression of anger or guilt. Participants' reactions were assessed after their partner for a collaborative task expressed anger or guilt due to failure on the task. Analysis indicated that participants inferred the attribution of responsibility made by the partner based on the expressed emotions; anger elicited an inference that the partner attributed the failure to the participant, while guilt led to an inference that the partner attributed the failure to the self. Further, expressers of anger were perceived as having more negative traits and were avoided more than those who expressed guilt. Discussion considered negative interpersonal effect of an expression of anger.


Implementation Strategies Used by States to Support Physical Activity Licensing Standards for Toddlers in Early Care and Education Settings: An Exploratory Qualitative Study.
Child care settings have been a target for childhood obesity prevention efforts; changing licensing standards to require physical activity (PA)-related best practices is one strategy being used by states. The mechanisms used to support implementation by providers remain understudied, despite the potential for these efforts to significantly impact child-level outcomes. This qualitative study explored the mechanisms used and any outstanding challenges to implementation experienced by US states with one specific best practice PA licensing standard for toddlers (60-90 min of PA/day). All states with the selected PA licensing standard were invited to participate (N = 9). Interviews were conducted with the licensing administrator and others in the state involved with implementation and referred by the Administrator (e.g., Licensing Inspectors). A total of 20 individuals from seven states were interviewed. All interviews were recorded, professionally transcribed, and analyzed to identify themes. A range of strategies was reported, however the majority of interviewees reported that providers experienced few challenges with implementation. Steps taken during the rulemaking process and technical assistance provided by licensing staff, government and nonprofit partners were frequently mentioned as contributing to implementation success. Challenges faced by licensing staff included lack of resources and a need for specific, age-appropriate PA-promoting activities that providers could easily implement. Based on the experience of participating states, other jurisdictions may want to explore adding PA-related requirements to their licensing standards. The lessons learned from these seven states may be helpful during the rulemaking and implementation processes.


Radioimmunological determination of thromboxane release in cardiac anaphylaxis.
A sensitive and specific radioimmunoassay for thromboxane B2 were detected in perfusates of anaphylactic guinea pig hearts. Indomethacin decreased thromboxane B2 levels in the perfusates to below the detection limit of the radioimmunoassay and concomitantly delayed the onset of coronary vasoconstriction after antigenic challenge.


[Intra-articular administration of galactosaminoglucuronoglycan sulfate in moderate primary gonarthritis. A pilot study].
The present study has been carried out in order to assess the clinical and functional efficacy and tolerability of galactosa-minoglucuronoglycan sulphate (GGGS). The molecule has been intra-articularly administered in patients affected by moderate primary gonarthritis, according to clinical and functional parameters. At the beginning and at the end of each period of treatment the following parameters have been evaluated: spontaneous pain, provoked pain, pain during active movement, pain during passive movement, pain at rising (7-8:00 a.m.) pain at bed time (10:00 p.m.), execution time (expressed in seconds) of up-down from a chair (five times), execution time (expressed in seconds) to walk 15 meters. Fifty patients affected by osteoarthritis of the knee were treated weekly, during one year, for two periods of six weeks each with a total amount of 12 injections. Treatment with Galactosaminglucuronoglycan sulphate (GGGS) seems to be effective in the therapy of osteoarthritis of the knee. The pain scores decreased and the functional levels significantly (p < 0.01) improved in a large proportion of patients and GGGS was very well tolerated. This study suggests that intra-articular treatment is a substantially new way for GGGS administration in the therapy of osteoarthritis of the knee.


Source derivation: application to topographic mapping of visual evoked potentials.
The application of a source derivation technique to topographic mapping of the pattern-reversal visual evoked potential is described. Comparisons were made with non-cephalic and common average reference recordings. Source derivation yielded wave forms which were less influenced by activity of reference origin. Source derivation wave forms were similar to those obtained with common average referencing but contour plots showed less spread of regions of maximal activity in the case of source derivation. It is suggested that source derivation may be useful in further studies of the topography and sources of cerebral evoked potentials.


Right ventricular recovery after cervical extra corporeal membrane oxygenation in a four-month-old male leading to left Berlin Heart Excor assistance.
A 4-month-old male with cardiogenic shock was implanted with cervical extracorporeal membrane oxygenation (ECMO). He had a biventricular heart failure associated to multiple organ failure (MOF). Right ventricular (RV) function and MOF recovered during five days ECMO leading to left ventricular assistance device (LVAD) implantation. This case shows the advantages of the double bridge strategy providing intervention time for complications refractory to VAD implantation and evaluation time of right and left ventricular function for potential mono- or bi-VAD implantation. Furthermore, there are no reports describing this strategy in young children leading to RV function recovery and its advantage.


Ultrastructure of botryoid sarcoma of the common bile duct.
A case of botryoid sarcoma of the common bile duct in a 4-year-old girl was reported. Electron microscopic examination disclosed that the neoplasm consisted of three types of cells: polygonal, elongated, and small cells. The former two contained moderate to large amounts of poorly developed myofibriles in the cytoplasm with occasional A, I and Z-bands. The small cells contained mitochondria and dilated rough endoplasmic reticulum but few myofilaments. Deposits of glycogen granules were constant components of the neoplastic cells. Mitosis was striking in the small cells. Abnormal multilaminar endoplasmic reticulum was observed in the small cells in the mitotic stage.


A comparison of the development of body components in Sudan desert sheep and goats.
Sixty-six male Sudan desert sheep and a similar number of Sudan desert goats representing different weight classes of sheep and goats commonly slaughtered in the local market were killed and carcass data collected. The patterns of the relative increase in weight of the components of the body were similar in both species with the exception of the skin, tail, liver and loin cut. At 30 kg empty bodyweight the goat yields a heavier carcass, omentum and head but a lighter skin, tail and feet than sheep. At 15 kg carcass weight goats possess less meat in the leg and plate cuts, but more meat in the loin and shoulder cuts.


Chronic administration of valproic acid reduces brain NMDA signaling via arachidonic acid in unanesthetized rats.
Evidence that brain glutamatergic activity is pathologically elevated in bipolar disorder suggests that mood stabilizers are therapeutic in the disease in part by downregulating glutamatergic activity. Such activity can involve the second messenger, arachidonic acid (AA, 20:4n - 6). We tested this hypothesis with regard to valproic acid (VPA), when stimulating glutamatergic N-methyl-D: -aspartate (NMDA) receptors in rat brain and measuring AA and related responses. An acute subconvulsant dose of NMDA (25 mg/kg i.p.) or saline was administered to unanesthetized rats that had been treated i.p. daily with VPA (200 mg/kg) or vehicle for 30 days. Quantitative autoradiography following intravenous [1-(14)C]AA infusion was used to image regional brain AA incorporation coefficients k*, markers of AA signaling. In chronic vehicle-pretreated rats, NMDA compared with saline significantly increased k* in 41 of 82 examined brain regions, many of which have high NMDA receptor densities, and also increased brain concentrations of the AA metabolites, prostaglandin E(2) (PGE(2)) and thromboxane B(2) (TXB(2)). VPA pretreatment reduced baseline concentrations of PGE(2) and TXB(2), and blocked the NMDA induced increases in k* and in eicosanoid concentrations. These results, taken with evidence that carbamazepine and lithium also block k* responses to NMDA in rat brain, suggest that mood stabilizers act in bipolar disorder in part by downregulating glutamatergic signaling involving AA.


Structural basis for diamide modulation of ryanodine receptor.
The diamide insecticide class is one of the top-selling insecticides globally. They are used to control a wide range of pests by targeting their ryanodine receptors (RyRs). Here, we report the highest-resolution cryo-electron microscopy (cryo-EM) structure of RyR1 in the open state, in complex with the anthranilic diamide chlorantraniliprole (CHL). The 3.2-Å local resolution map facilitates unambiguous assignment of the CHL binding site. The molecule induces a conformational change by affecting the S4-S5 linker, triggering channel opening. The binding site is further corroborated by mutagenesis data, which reveal how diamide insecticides are selective to the Lepidoptera group of insects over honeybee or mammalian RyRs. Our data reveal that several pests have developed resistance via two mechanisms, steric hindrance and loss of contact. Our results provide a foundation for the development of highly selective pesticides aimed at overcoming resistance and therapeutic molecules to treat human myopathies.


SH3-domain mutations selectively disrupt Csk homodimerization or PTPN22 binding.
The kinase Csk is the primary negative regulator of the Src-family kinases (SFKs, e.g., Lck, Fyn, Lyn, Hck, Fgr, Blk, Yes), phosphorylating a tyrosine on the SFK C-terminal tail that mediates autoinhibition. Csk also binds phosphatases, including PTPN12 (PTP-PEST) and immune-cell PTPN22 (LYP/Pep), which dephosphorylate the SFK activation loop to promote autoinhibition. Csk-binding proteins (e.g., CBP/PAG1) oligomerize within membrane microdomains, and high local concentration promotes Csk function. Purified Csk homodimerizes in solution through an interface that overlaps the phosphatase binding footprint. Here we demonstrate that Csk can homodimerize in Jurkat T cells, in competition with PTPN22 binding. We designed SH3-domain mutations in Csk that selectively impair homodimerization (H21I) or PTPN22 binding (K43D) and verified their kinase activity in solution. Disruption of either interaction in cells, however, decreased the negative-regulatory function of Csk. Csk W47A, a substitution previously reported to block PTPN22 binding, had a secondary effect of impairing homodimerization. Csk H21I and K43D will be useful tools for dissecting the protein-specific drivers of autoimmunity mediated by the human polymorphism PTPN22 R620W, which impairs interaction with Csk and with the E3 ubiquitin ligase TRAF3. Future investigations of Csk homodimer activity and phosphatase interactions may reveal new facets of SFK regulation in hematopoietic and non-hematopoietic cells.


Effects of polysaccharide intercellular adhesin (PIA) in an ex vivo model of whole blood killing and in prosthetic joint infection (PJI): A role for C5a.
A major complication of using medical devices is the development of biofilm-associated infection caused by Staphylococcus epidermidis where polysaccharide intercellular adhesin (PIA) is a major mechanism of biofilm accumulation. PIA affects innate and humoral immunity in isolated cells and animal models. Few studies have examined these effects in prosthetic joint infection (PJI). This study used ex vivo whole blood modelling in controls together with matched-serum and staphylococcal isolates from patients with PJI. Whole blood killing of PIA positive S. epidermidis and its isogenic negative mutant was identical. Differences were unmasked in immunosuppressed whole blood pre-treated with dexamethasone where PIA positive bacteria showed a more resistant phenotype. PIA expression was identified in three unique patterns associated with bacteria and leukocytes, implicating a soluble form of PIA. Purified PIA reduced whole blood killing while increasing C5a levels. In clinically relevant staphylococcal isolates and serum samples from PJI patients; firstly complement C5a was increased 3-fold compared to controls; secondly, the C5a levels were significantly higher in serum from PJI patients whose isolates preferentially formed PIA-associated biofilms. These data demonstrate for the first time that the biological effects of PIA are mediated through C5a in patients with PJI.


Dynamics of Ostertagia spp. and Cooperia oncophora in field-grazed cattle from weaning to 2 years old in New Zealand, with particular reference to arrested development.
Gastrointestinal nematode parasite burdens were monitored in a herd of field-grazed cattle from weaning to 2 years old to allow observations to be made on the dynamics of burdens of arrested Ostertagia spp. and Cooperia oncophora. Arrested Ostertagia spp. accumulated in the herd over their first autumn and winter to reach a peak in late winter/spring (August-October). From October until late January they declined at a rate comparable to that at which they had accumulated. Numbers continued to decline at a slower rate until the following autumn (May) when they began to increase again. Observations on the decline of worm burdens in a group of animals which had been moved from pasture to worm-free conditions in October indicated that intake of infective larvae over late spring/summer contributed little to the number of arrested Ostertagia spp. in the herd. No clinical Type II ostertagiosis was observed in the field-grazed animals over the time of decline of the arrested worm burden, nor was there an increase in the number of adult Ostertagia spp. present. However, two of the animals removed from pasture to worm-free conditions developed Type II ostertagiosis approximately a month after the move. Worm burdens of successive groups of "tracers" grazed with the herd every 2 months indicated that there was a small seasonal increase in the propensity of Ostertagia spp. for arrested development. However, estimated accumulation rates of arrested larvae calculated from tracer worm burdens, suggest that this seasonal increase in propensity was not adequate in itself to account for the accumulation rate observed in "resident" animals of the herd. No comparable accumulation of arrested C. oncophora took place in resident animals over the first autumn and winter of the trial despite the fact that tracer worm burdens over this period indicated a marked seasonal increase in the propensity for arrested development by this species.


Diet and genetics: Trp-ing over food sensitivity.
Laboratory-reared Caenorhabditis elegans eat Escherichia coli. A new study demonstrates a strong diet-gene interaction: worms with reduced nhr-114 activity are fertile when fed E. coli K-12 strains but are sterile on E. coli B. Surprisingly, tryptophan supplementation of E. coli B restores worm fertility.


Osteoporotic vertebral compression fractures: a rare complication of radiotherapy in a patient with lung cancer.
The development of bone fractures after radiotherapy is a rare event which mainly concerns the pelvis or the long bones. This complication is unusual in the vertebrae. We describe the case of a 66-year-old male patient with lung cancer who was treated with combined radio-chemotherapy and developed dorsal pain secondary to vertebral compression 4 months after the end of radiotherapy. Investigations led to a diagnosis of post-radiotherapy vertebral osteonecrosis. It is important to differentiate metastatic lesions from radiological complications. It is not possible to differentiate a metastasis from a recent osteoporotic compression fracture by imaging. A bone biopsy may therefore be necessary. Metastatic bone involvement is common in patients with lung cancer. When images are not typical of secondary progression, however, and there is no change in the general state of the patient, evidence of thoracic progression of the tumour or distal progression other than bone, vertebral osteoporotic complications should be considered. It is important that a wrong diagnosis is not made without histological proof of metastasis which has a poor prognosis.


US132 Cyclodextrin Glucanotransferase Engineering by Random Mutagenesis for an Anti-Staling Purpose.
The use of the cyclodextrin glucanotransferase (CGTase) of the US132 strain, which is an effective anti-staling agent, has been hampered by its high cyclization activity. Since that random mutagenesis using error-prone PCR is nowadays a method of choice for enzymes engineering, we have optimized this method by adjusting manganese concentration in order to obtain a high percentage of active CGTase mutants. Therefore, the amplification of the gene encoding the US132 CGTase was performed using a MnCl2 concentration ranging between 0 and 0.5 mM. The finding showed that a manganese concentration of 0.04 mM allowed for 90 % of active mutants. A simple method to rapidly screen the obtained mutants was also developed. After the examination of a small library (of less than 1000 clones), the active mutant named MJ13 was selected for a significant decrease in the cyclization activity, thereby showing a remarkable change in the enzyme specificity towards starch dextrinizing. Sequence analysis showed that MJ13 is a triple mutant with two mutations in the catalytic domain (K47E and S382P) and one substitution in the starch binding domain (N655S).


Selectin-deficiency reduces the number of spontaneous metastases in a xenograft model of human breast cancer.
Metastasis formation is a complex process still poorly understood. Previous work in a colon cancer xenograft model showed that E(ndothelial) and P(latelet) selectins mediate spontaneous metastasis to the lungs. To investigate the functional role of selectins in breast cancer, human DU4475 breast cancer cells were injected subcutaneously into pfp-/-rag2-/- mice and in all their selectin-deficient variants (EP-/-, E-/- and P-/-). Pfp-/-rag2-/- mice as well as all their selectin-deficient variants developed primary tumours and spontaneous metastases. Compared with the wild-type mice, disseminated tumours cells were significantly lower (74% reduction, P=0.046) in the bone marrow of selectin-deficient mice. Pfp-/-rag2-/- mice developed significantly higher numbers of lung metastases (6644.83±741.77) than the E-/- (4053.33±112.58; P=0.002) and the EP-/- pfp-/-rag2-/- mice (4665.65±754.50; P<0.001). The results indicate that E- and P-selectins play a role in spontaneous metastasis formation both into bone marrow and lungs. However, spontaneous metastasis was not completely abrogated, hence additional cell adhesion molecules must be involved in the metastatic spread.


Protective mechanisms of 10-gingerol against myocardial ischemia may involve activation of JAK2/STAT3 pathway and regulation of Ca2+ homeostasis.
10-Gingerol (10-Gin), an active ingredient extracted from ginger, has been reported to have beneficial effects on the cardiovascular system. However, its protective effects on myocardial ischemia (MI) and the underlying cellular mechanisms are still unclear. To investigate the protection conferred by 10-Gin against MI injury and its potential mechanisms in cardiomyocytes via patch-clamp and molecular biology techniques. A rat MI model was established using the subcutaneous injection of isoproterenol (85 mg/kg) administered on two consecutive days. 10-Gin was pre-administered to rats for seven days to assess its cardio-protection. The patch-clamp and IonOptix Myocam detection techniques were used to investigated 10-Gin's effects on L-type Ca2+ channels (LTCCs), Ca2+ transients and cell contractility in isolated rat cardiomyocytes. 10-Gin administration alleviated MI injury, improved cardiac function and myocardial histopathology, reduced myocardial infarct area, downregulated oxidative stress and Ca2+ levels, and decreased the expression of apoptotic factors. Importantly, 10-Gin led to an increase in phosphorylated Janus kinase 2 and signal transducer and activator of transcription 3 (JAK2 and STAT3, respectively) expressions. Furthermore, 10-Gin inhibited LTCCs in a concentration-dependent manner with a half-maximal inhibitory concentration of 75.96 μM. Moreover, 10-Gin administration inhibited Ca2+ transients and cell contractility. Our results suggest that 10-Gin exerts cardioprotective effects on MI in vivo and in vitro in connection with the inhibition of oxidative stress and apoptosis via activation of the JAK2/STAT3 signalling pathway, and regulation of Ca2+ homeostasis by LTCCs.


The correlation between PM2.5 exposure and hypertensive disorders in pregnancy: A Meta-analysis.
To find the correlation between exposure to PM2.5 (fine particulate matter) and hypertensive disorders in pregnancy (HDP), and provide medical evidence for decreasing the incidence of hypertensive disorders in pregnancy. A combination of computer and manual retrieval was used to search for keywords in PubMed (385 records), Cochrane Library (20 records), Web of Science (419 records) and Embase (325 records). Finally, ten epidemiological articles were considered in this meta-analysis. Stata 13.0 was used to examine the heterogeneity among the studies and to calculate the combined effect value (OR, odds ratio) by selecting the corresponding models. Sensitivity analysis and publication bias test were also performed. Meta-analysis indicated that there was an association between PM2.5 exposure (per 10 µg/m3 increase) and hypertensive disorders in pregnancy (OR = 1.52, 95% CI: 1.24-1.87). Exposure to PM2.5 (per 10 µg/m3 increase) enhanced the risk of pre-eclampsia (OR = 1.31, 95% CI: 1.07-1.61), but there was no evidence relating exposure to PM2.5 to gestational hypertension (OR = 1.35, 95% CI: 0.98-1.87). There is a significant link between exposure to PM2.5 and hypertensive disorders in pregnancy. The first and the third trimester were more susceptible to PM2.5 exposure. It is recommended to further strengthen protective measures against PM2.5 during pregnancy.


Fixed-time periodic stabilization of discontinuous reaction-diffusion Cohen-Grossberg neural networks.
This paper aims to study the fixed-time stabilization of a class of delayed discontinuous reaction-diffusion Cohen-Grossberg neural networks. Firstly, by providing some relaxed conditions containing indefinite functions and based on inequality techniques, a new fixed-time stability lemma is given, which can improve the traditional ones. Secondly, based on state-dependent switching laws, the periodic wave solution of the formulated networks is transformed into the periodic solution of ordinary differential system. By utilizing differential inclusions theory and coincidence theorem, the existence of periodic solutions is obtained. Thirdly, based on the new fixed-time stability lemma, the periodic solutions are stabilized at zero in a fixed-time, which is a new topic on reaction-diffusion networks. Moreover, the established criteria are all delay-dependent, which are less conservative than the previous delay-independent ones for ensuring the stabilization of delayed reaction-diffusion networks. Finally, two examples give numerical explanations of the proposed results and highlight the influence of delays.


In private practice, informed consent is interpreted as providing explanations rather than offering choices: a qualitative study.
How do physiotherapists working in private practice understand and interpret the meaning and significance of informed consent in everyday clinical practice? Qualitative study using semi-structured interviews. Seventeen physiotherapists purposefully recruited from metropolitan private practices where treatment was on a one-on-one basis. Therapists defined informed consent as an implicit component of their routine clinical explanations, rather than a process of providing explicit patient choices. Therapists' primary concern was to provide information that led to a (therapist-determined) beneficial therapeutic outcome, rather than to enhance autonomous patient choice. Explicit patient choice and explicit informed consent were defined as important only if patients requested information or therapists recognised risks associated with the treatment. Physiotherapists defined informed consent within a context of achieving therapeutic outcomes rather than a context of respect for patient autonomy and autonomous choice. Physiotherapy practice guidelines developed to ensure compliance with ethical and legal obligations may therefore be followed only if they fit with therapists' understanding and interpretation of a desired therapeutic outcome.


Hepatitis E virus genotype 4, Nanjing, China, 2001-2011.
During 2001-2011, hepatitis E virus (HEV) was found in the blood of patients in Nanjing, China. All HEV-positive patients had virus genotype 4; subgenotype 4a was predominant. The effective population of HEV in Nanjing increased in ≈1980 and continued until ≈2003 when it plateaued.


A Computational Pipeline for Predicting Cancer Neoepitopes.
Mutant Peptide eXtractor and Informer (MuPeXI), by Bjerregaard et al. (Cancer Immunol Immunother CII 66:1123-1130, 2017), is a program which identifies tumor-specific peptides and assesses their potential to be neoepitopes. MuPeXI takes as input a VCF file and a list of human leukocyte antigen (HLA) types and optionally a gene expression profile to assess a peptide's potential to be a neoepitope. MuPeXI can be downloaded and run both locally and on a web server. Here, we describe a pipeline for processing the input data so that it can be used for MuPeXI and how to run MuPeXI both locally and as a web server.


[Celiac disease associated with insulin-dependent diabetes mellitus: report of a case].
We report the association of diabetes mellitus and celiac disease in one patient, which was confirmed by intestinal biopsy, the withdraw of gluten from the diet, and by a positive challenge test. We emphasize the importance to recognize this association to improve the clinical management of patients with diabetes mellitus.


Sustained antimicrobial activity and reduced toxicity of oxidative biocides through biodegradable microparticles.
The spread of antibiotic-resistant pathogens requires new treatments. Small molecule precursor compounds that produce oxidative biocides with well-established antimicrobial properties could provide a range of new therapeutic products to combat resistant infections. The aim of this study was to investigate a novel biomaterials-based approach for the manufacture, targeted delivery and controlled release of a peroxygen donor (sodium percarbonate) combined with an acetyl donor (tetraacetylethylenediamine) to deliver local antimicrobial activity via a dynamic equilibrium mixture of hydrogen peroxide and peracetic acid. Entrapment of the pre-cursor compounds into hierarchically structured degradable microparticles was achieved using an innovative dry manufacturing process involving thermally induced phase separation (TIPS) that circumvented compound decomposition associated with conventional microparticle manufacture. The microparticles provided controlled release of hydrogen peroxide and peracetic acid that led to rapid and sustained killing of multiple drug-resistant organisms (methicillin-resistant Staphylococcus aureus and carbapenem-resistant Escherichia coli) without associated cytotoxicity in vitro nor intracutaneous reactivity in vivo. The results from this study demonstrate for the first time that microparticles loaded with acetyl and peroxygen donors retain their antimicrobial activity whilst eliciting no host toxicity. In doing so, it overcomes the detrimental effects that have prevented oxidative biocides from being used as alternatives to conventional antibiotics. The manuscript explores a novel approach to utilize the antimicrobial activity of oxidative species for sustained killing of multiple drug-resistant organisms without causing collateral tissue damage. The results demonstrate, for the first time, the ability to load pre-cursor compounds into porous polymeric structures that results in their release and conversion into oxidative species in a controlled manner. Until now, the use of oxidative species has not been considered as a candidate therapeutic replacement for conventional antibiotics due to difficulties associated with handling during manufacture and controlling sustained release without causing undesirable tissue damage. The ultimate impact of the research could be the creation of new materials-based anti-infective chemotherapeutic agents that have minimal potential for giving rise to antimicrobial resistance.


[Biliary tract surgery in childhood].
In contrast to gallstones, which are increasingly diagnosed in children and adolescents, congenital or perinatally acquired malformations of the biliary tree are rare entities. The surgical treatment of these diseases frequently represents a technical challenge. For children with biliary atresia Kasai portoenterostomy is a therapeutic option in the first weeks of life. Most affected children, however, require a liver transplantation before they enter adulthood. Segmental dilatations of the bile duct carry the risk of ascending infections and malignant transformation. This requires early resection of the biliary tree and reconstruction with a bilioenteric anastomosis. While laparoscopic cholecystectomy has become the standard approach also for children, the value of minimally invasive procedures for more complex surgical procedures of the biliary tract has not yet been defined.


The role of peripheral blood eosinophil counts in COVID-19 patients.
Coronavirus disease 2019 (COVID-19) emerged in Wuhan city and rapidly spread globally outside China. We aimed to investigate the role of peripheral blood eosinophil (EOS) as a marker in the course of the virus infection to improve the efficiency of diagnosis and evaluation of COVID-19 patients. 227 pneumonia patients who visited the fever clinics in Shanghai General Hospital and 97 hospitalized COVID-19 patients admitted to Shanghai Public Health Clinical Center were involved in a retrospective research study. Clinical, laboratory, and radiologic data were collected. The trend of EOS level in COVID-19 patients and comparison among patients with different severity were summarized. The majority of COVID-19 patients (71.7%) had a decrease in circulating EOS counts, which was significantly more frequent than other types of pneumonia patients. EOS counts had good value for COVID-19 prediction, even higher when combined with neutrophil-to-lymphocyte ratio. Patients with low EOS counts at admission were more likely to have fever, fatigue, and shortness of breath, with more lesions in chest CT and radiographic aggravation, and longer length of hospital stay and course of disease than those with normal EOS counts. Circulating EOS level gradually increased over the time, and was synchronous with the improvement in chest CT (12 days vs 13 days, P = .07), later than that of body temperature (12 days vs 10 days, P = .014), but earlier than that of the negative conversion of nucleic acid assays (12 days vs 17 days, P = .001). Peripheral blood EOS counts may be an effective and efficient indicator in diagnosis, Evaluation, and prognosis monitoring of COVID-19 patients.


Confirmation of diosmetin 3-O-glucuronide as major metabolite of diosmin in humans, using micro-liquid-chromatography-mass spectrometry and ion mobility mass spectrometry.
Diosmin is a flavonoid often administered in the treatment of chronic venous insufficiency, hemorrhoids, and related affections. Diosmin is rapidly hydrolized in the intestine to its aglicone, diosmetin, which is further metabolized to conjugates. In this study, the development and validations of three new methods for the determination of diosmetin, free and after enzymatic deconjugation, and of its potential glucuronide metabolites, diosmetin-3-O-glucuronide, diosmetin-7-O-glucuronide, and diosmetin-3,7-O-glucuronide from human plasma and urine are presented. First, the quantification of diosmetin, free and after deconjugation, was carried out by high-performance liquid chromatography coupled with tandem mass spectrometry, on an Ascentis RP-Amide column (150 × 2.1 mm, 5 μm), in reversed-phase conditions, after enzymatic digestion. Then glucuronide metabolites from plasma were separated by micro-liquid chromatography coupled with tandem mass spectrometry on a HALO C18 (50 × 0.3 mm, 2.7 μm, 90 Å) column, after solid-phase extraction. Finally, glucuronides from urine were measured using a Discovery HSF5 (100 × 2.1 mm, 5 μm) column, after simple dilution with mobile phase. The methods were validated by assessing linearity, accuracy, precision, low limit of quantification, selectivity, extraction recovery, stability, and matrix effects; results in agreement with regulatory (Food and Drug Administration and European Medicines Agency) guidelines acceptance criteria were obtained in all cases. The methods were applied to a pharmacokinetic study with diosmin (450 mg orally administered tablets). The mean C max of diosmetin in plasma was 6,049.3 ± 5,548.6 pg/mL. A very good correlation between measured diosmetin and glucuronide metabolites concentrations was obtained. Diosmetin-3-O-glucuronide was identified as a major circulating metabolite of diosmetin in plasma and in urine, and this finding was confirmed by supplementary experiments with differential ion-mobility mass spectrometry.


[Coinfections in contact lens-associated mycotic keratitis with Pseudomonas or Acanthamoeba].
Contact lens-associated keratitis is becoming increasingly more frequent. Fungal keratitis is a relatively rare clinical picture but must be taken very seriously. Especially in the early stages of the disease, it may be clinically misdiagnosed and adequate treatment is therefore delayed. In treatment-resistant contact lens-associated fungal keratitis, coinfections or superinfections can occur. We present two patients with an initially unclear keratitis, in whom a fungal keratitis with coinfection of Pseudomonas aeruginosa and Acanthamoeba, respectively, could be confirmed. In both cases an urgent excimer laser penetrating keratoplasty with interrupted sutures and adequate local topical treatment for 8 weeks was successful.


Survival and growth of Enterobacter sakazakii in infant rice cereal reconstituted with water, milk, liquid infant formula, or apple juice.
To determine survival and growth characteristics of Enterobacter sakazakii in infant rice cereal as affected by type of liquid used for reconstitution and storage temperature after reconstitution. A commercially manufactured dry infant rice cereal was reconstituted with water, apple juice, milk, or liquid infant formula, inoculated with a 10-strain mixture of E. sakazakii at populations of 0.27, 0.93, and 9.3 CFU ml(-1), and incubated at 4, 12, 21 or 30 degrees C for up to 72 h. Growth did not occur in cereal reconstituted with apple juice, regardless of storage temperature, or in cereal reconstituted with water, milk, or formula and stored at 4 degrees C. The lag time for growth in cereal reconstituted with water, milk, or formula was decreased as the incubation temperature (12, 21 and 30 degrees C) was increased. Upon reaching maximum populations of 7-8 log10 CFU ml(-1), in some instances populations decreased to nondetectable levels during subsequent storage which was concurrent with decreases in pH. Enterobacter sakazakii initially at very low populations can rapidly grow in infant rice cereal reconstituted with water, milk, or infant formula. Reconstituted infant rice cereal can support luxuriant growth of E. sakazakii. Reconstituted cereal that is not immediately consumed should be discarded or stored at a temperature at which E. sakazakii and other food-borne pathogens cannot grow.


Calcium signaling in closely related protozoan groups (Alveolata): non-parasitic ciliates (Paramecium, Tetrahymena) vs. parasitic Apicomplexa (Plasmodium, Toxoplasma).
The importance of Ca2+-signaling for many subcellular processes is well established in higher eukaryotes, whereas information about protozoa is restricted. Recent genome analyses have stimulated such work also with Alveolates, such as ciliates (Paramecium, Tetrahymena) and their pathogenic close relatives, the Apicomplexa (Plasmodium, Toxoplasma). Here we compare Ca2+ signaling in the two closely related groups. Acidic Ca2+ stores have been characterized in detail in Apicomplexa, but hardly in ciliates. Two-pore channels engaged in Ca2+-release from acidic stores in higher eukaryotes have not been stingently characterized in either group. Both groups are endowed with plasma membrane- and endoplasmic reticulum-type Ca2+-ATPases (PMCA, SERCA), respectively. Only recently was it possible to identify in Paramecium a number of homologs of ryanodine and inositol 1,3,4-trisphosphate receptors (RyR, IP3R) and to localize them to widely different organelles participating in vesicle trafficking. For Apicomplexa, physiological experiments suggest the presence of related channels although their identity remains elusive. In Paramecium, IP3Rs are constitutively active in the contractile vacuole complex; RyR-related channels in alveolar sacs are activated during exocytosis stimulation, whereas in the parasites the homologous structure (inner membrane complex) may no longer function as a Ca2+ store. Scrutinized comparison of the two closely related protozoan phyla may stimulate further work and elucidate adaptation to parasitic life. See also "Conclusions" section.


Analysis of cumulative live birth rate outcomes of three ovarian stimulation protocols in patients after laparoscopic cystectomy of ovarial endometrioma: a retrospective cohort study.
Previous studies have reported that after laparoscopic cystectomy of ovarial endometrioma, the ovarian response to gonadotropin (Gn) significantly decreased. However, for patients with diminished ovarian reserve (DOR) after ovarian surgery, how to choose the most appropriate controlled ovarian hyperstimulation protocol has not been concluded. Compared with the traditional agonist regimen, the gonadotropin (Gn)-releasing hormone (GnRH) antagonist, microstimulation, and progestin-primed ovarian stimulation (PPOS) protocols are simple to operate and have a shorter cycle, which are often used in patients with DOR. So the purpose of our study is to compare the assisted reproductive outcomes of these three controlled ovarian hyperstimulation protocols in patients with DOR following laparoscopic cystectomy of ovarial endometrioma. In this retrospective cohort study, 89 patients with DOR who had undergone in vitro fertilisation/intracytoplasmic sperm injection at the Department of Reproductive Medicine at the Third Affiliated Hospital of Zhengzhou University from 1 to 2018 to 31 December 2020 were included. According to the controlled ovarian hyperstimulation protocols employed, the patients were divided into GnRH antagonist (38 patients), PPOS (27 patients), and microstimulation (24 patients) groups. The basic data and clinical outcomes of the three groups were compared. The main outcome measure was the cumulative live birth rate. No significant differences in the age of the female patients and their spouses and female patients' body mass index and basal endocrine levels (follicle-stimulating hormone and oestradiol) were noted among the three groups (P > 0.05). The GnRH antagonist group had higher antral follicle counts, greater endometrial thickness on the human chorionic Gn injection day, greater number of oocytes retrieved, and higher two pronuclear embryo counts than did the other two groups. However, the starting dosage of Gn was lower in the GnRH antagonist group than in the other two groups. The microstimulation group had a significantly higher oocyte output rate and high-quality embryo rate than did the other two groups (P < 0.05). No significant differences in the total dosage of Gn, cumulative pregnancy rate, cumulative live birth rate, viable embryo rate, and blastocyst formation rate were observed among the three groups (P > 0.05). In conclusion, for patients aged under 40 years who experienced DOR after laparoscopic cystectomy of ovarial endometrioma, GnRH antagonist protocol and PPOS protocol can obtain better ovulation induction outcomes and cumulative live birth rate than microstimulation protocol, and are more suitable ovulation induction protocols.


Production and applications of biobased packaging materials for the food industry.
Materials based on renewable resources are being developed at an increasing rate. Today, the only biobased food-packaging materials used commercially on a major scale are based on cellulose. However, materials based on proteins, starch, polylactate and other renewable resources may be the food-packaging materials of tomorrow. The paper presents some of the different biobased materials and their potential as food-packaging materials.


Uptake of a novel anticonvulsant compound, 2-amino-7-phosphono-[4,5-3H]heptanoic acid, into mouse brain.
The time course of radioactivity in plasma, liver and brain is described following the intraperitoneal injection of 2-amino-7-phosphono-[4,5-3H]heptanoic acid in mice. Using high performance liquid chromatography of dansylated extracts of mouse brain the radioactivity attributable to 2-amino-7-phosphono-[4,5-3H]heptanoic acid has been determined at 0.25-180 min after injection. The time course, with a peak concentration at 30 min, declining markedly between 90 and 180 min, corresponds to the anticonvulsant action.


Circulating tumor DNA guided adjuvant chemotherapy in stage II colon cancer (MEDOCC-CrEATE): study protocol for a trial within a cohort study.
Accurate detection of patients with minimal residual disease (MRD) after surgery for stage II colon cancer (CC) remains an urgent unmet clinical need to improve selection of patients who might benefit form adjuvant chemotherapy (ACT). Presence of circulating tumor DNA (ctDNA) is indicative for MRD and has high predictive value for recurrent disease. The MEDOCC-CrEATE trial investigates how many stage II CC patients with detectable ctDNA after surgery will accept ACT and whether ACT reduces the risk of recurrence in these patients. MEDOCC-CrEATE follows the 'trial within cohorts' (TwiCs) design. Patients with colorectal cancer (CRC) are included in the Prospective Dutch ColoRectal Cancer cohort (PLCRC) and give informed consent for collection of clinical data, tissue and blood samples, and consent for future randomization. MEDOCC-CrEATE is a subcohort within PLCRC consisting of 1320 stage II CC patients without indication for ACT according to current guidelines, who are randomized 1:1 into an experimental and a control arm. In the experimental arm, post-surgery blood samples and tissue are analyzed for tissue-informed detection of plasma ctDNA, using the PGDx elio™ platform. Patients with detectable ctDNA will be offered ACT consisting of 8 cycles of capecitabine plus oxaliplatin while patients without detectable ctDNA and patients in the control group will standard follow-up according to guideline. The primary endpoint is the proportion of patients receiving ACT when ctDNA is detectable after resection. The main secondary outcome is 2-year recurrence rate (RR), but also includes 5-year RR, disease free survival, overall survival, time to recurrence, quality of life and cost-effectiveness. Data will be analyzed by intention to treat. The MEDOCC-CrEATE trial will provide insight into the willingness of stage II CC patients to be treated with ACT guided by ctDNA biomarker testing and whether ACT will prevent recurrences in a high-risk population. Use of the TwiCs design provides the opportunity to randomize patients before ctDNA measurement, avoiding ethical dilemmas of ctDNA status disclosure in the control group. Netherlands Trial Register: NL6281/NTR6455 . Registered 18 May 2017, https://www.trialregister.nl/trial/6281.


Beyond NIMBYs and NOOMBYs: what can wind farm controversies teach us about public involvement in hospital closures?
Many policymakers, researchers and commentators argue that hospital closures are necessary as health systems adapt to new technological and financial contexts, and as population health needs in developed countries shift. However closures are often unpopular with local communities. Previous research has characterised public opposition as an obstacle to change. Public opposition to the siting of wind farms, often described as NIMBYism (Not In My Back Yard), is a useful comparator issue to the perceived NOOMBYism (Not Out Of My Back Yard) of hospital closure protestors. The analysis of public attitudes to wind farms has moved from a fairly crude characterisation of the 'attitude-behaviour gap' between publics who support the idea of wind energy, but oppose local wind farms, to empirical, often qualitative, studies of public perspectives. These have emphasised the complexity of public attitudes, and revealed some of the 'rational' concerns which lie beneath protests. Research has also explored processes of community engagement within the wind farm decision-making process, and the crucial role of trust between communities, authorities, and developers. Drawing on what has been learnt from studies of opposition to wind farms, we suggest a range of questions and approaches to explore public perspectives on hospital closure more thoroughly. Understanding the range of public responses to service change is an important first step in resolving the practical dilemma of effecting health system transformation in a democratic fashion.


Dose dependency of earthworm powder on antithrombotic and fibrinolytic effects.
The freeze-dried powder of Lumbricus rubellus earthworm was administered orally to rats and its fibrinolytic and antithrombotic effects were investigated. The fibrinolytic activity of plasma was determined by measuring the plasmin activity of the euglobulin fraction and was increased to two-folds of the control at a dose of 0.5 g/kg/day and five times with 1 g/kg/day after 4-day administration. The antithrombotic effect was studied in an arterio-venous shunt model of rats. The thrombus weight decreased significantly from 43.2 mg to 32.4 mg at a dose of 0.5 g/kg/day after 8-day treatment. The level of fibrinogen/fibrin degradation product (FDP) in serum was elevated in a dose-dependent manner during the treatment period. On the 8th day after administration, the FDP value was increased to 7.7 micrograms/ml compared with the control value of 3.3 micrograms/ml. These results support that earthworm powder is valuable for the prevention and/or treatment of thrombotic conditions.


Developing a framework for a western Canadian companion animal surveillance initiative: Case definitions and the role of the veterinarian.
Surveillance data on companion animals in western Canada are extremely limited. Previous research by the principal investigators established a list of potential canine pathogens of relevance to public health for inclusion in the Western Canadian Companion Animal Surveillance Initiative (CASI). Our objective was to assess veterinary interest in contributing to companion animal surveillance, and to gather baseline data on specific canine pathogens of interest to create surveillance-specific case definitions. An invitation to participate in an online survey was disseminated to all clinical veterinarians across the provinces of Alberta, Saskatchewan, and Manitoba. There was a moderate level of interest (median: 7.5/10) from veterinarians to participate in the surveillance of companion animals. The majority (85%, 51/60) of veterinarians participating in the survey recorded diagnosing at least 1 of the pathogens of interest over a 5-year interval. Based on survey responses, several surveillance case definitions were formulated for pathogen groups of interest, most of which require laboratory testing for confirmation. This study identified the willingness, practicality, and importance of veterinarians or veterinary clinics participating in companion animal surveillance.


Prediction of serious complications in patients with pulmonary thromboembolism and solid cancer: Validation of the EPIPHANY Index in a prospective cohort of patients from the PERSEO study.
There is currently no validated score capable of classifying cancer-associated pulmonary embolism (PE) in its full spectrum of severity. This study has validated the EPIPHANY Index, a new tool to predict serious complications in cancer patients with suspected or unsuspected PE. The PERSEO Study prospectively recruited individuals with PE and active cancer or receiving antineoplastic therapy from 22 Spanish hospitals. The estimation of the relative frequency θ of complications based on the EPIPHANY Index categories was made using the Bayesian alternative for the binomial test. A total of 900 patients, who were diagnosed with PE between October 2017 and January 2020, were enrolled. The rate of serious complications at 15 days was 11.8%, 95% highest density interval [HDI], 9.8-14.1%. Of the EPIPHANY low-risk patients, 2.4% (95% HDI, 0.8-4.6%) had serious complications, as did 5.5% (95% HDI, 2.9-8.7%) of the moderate-risk participants and 21.0% (95% HDI, 17.0-24.0%) of those with high-risk episodes. The EPIPHANY Index was associated with overall survival (OS) in patients with different risk levels: median OS was 16.5, 14.4, and 4.4 months for those at low, intermediate, and high risk, respectively. Both the EPIPHANY Index and the Hestia criteria exhibited greater negative predictive value and a lower negative likelihood ratio than the remaining models. The incidence of bleeding at 6 months was 6.2% (95% HDI, 2.9-9.5%) in low/moderate-risk vs 12.7% (95% HDI, 10.1-15.4%) in high-risk (p-value = 0.037) episodes. Of the outpatients, serious complications at 15 days were recorded in 2.1% (95% HDI, 0.7-4.0%) of the cases with EPIPHANY low/intermediate-risk vs 5.3% (95% HDI, 1.7-11.8%) in high-risk cases. We have validated the EPIPHANY Index in patients with incidental or symptomatic cancer-related PE. This model can contribute to standardize decision-making in a scenario lacking quality evidence.


Evaluation of sternal bone marrow aspiration for detection of tumor cells in patients with bronchogenic carcinoma.
We evaluated sternal bone marrow aspiration as a routine pretreatment procedure in patients with bronchogenic carcinoma. The overall rate of identification of tumor cells by this technique was found to be low. When oat cell bronchogenic carcinoma was considered as a separate entity, the positive rate was higher. However, in no instance did the bone marrow aspiration result in a change in clinical staging.


[Cardiac contractility modulation. A new form of therapy for patients with heart failure and narrow QRS complex?].
Cardiac contractility modulation (CCM) is a stimulation therapy by an implantable impulse generator, which enhances ventricular contractile performance by delivering CCM impulses to the right ventricle during the absolute refractory period. The CCM signals mediate increased inotropy by prolonging the duration of the action potential, which leads to an enhanced influx of calcium into cardiomyocytes and a greater release of calcium by the sarcoplasmic reticulum. The increase of cardiac contractility is not associated with increased oxygen consumption. Several small studies have shown that CCM therapy can safely improve symptoms of heart failure and peak oxygen consumption in patients with moderate to severe heart failure who are not eligible for resynchronization therapy. Therefore, CCM is a novel potential therapy for patients with heart failure, an ejection fraction ≤ 35 % and a normal QRS duration < 130 ms. However, apart from selecting appropriate patients for CCM therapy there are still unanswered questions, such as the impact of CCM therapy on established clinical endpoints. At present no data are available which have shown that CCM therapy leads to reduction of hospitalization for heart failure or mortality.