Adaptive Evolution of CENP-A in Percid Fishes.
Centromeric protein A (CENP-A) is the epigenetic determinant of centromeres. This protein has been shown to be adaptively evolving in a number of animal and plant species. In a previous communication we were able to demonstrate that signs of adaptive evolution were detected in the comparison of CENP-A sequences from three percid fish species. In this study we isolated the CENP-A gene from eight additional species from the Percidae family. With these sequences and those previously obtained, we carried out a more robust statistical analysis of codon specific positive selection in CENP-A coding sequences of eleven percid species. We were able to demonstrate that at least two amino acid positions within the N-terminal tail are under strong positive selection and that one of these positions is potentially a substrate for phosphorylation. While nonsynonymous substitutions were detected in the histone fold domain, these were not statistically supported as resulting from positive selection.


Soybean cultivar resistant to Mungbean Yellow Mosaic India Virus infection induces viral RNA degradation earlier than the susceptible cultivar.
Yellow mosaic disease caused by whitefly-transmitted bipartite Geminiviruses is one of the major constraints on productivity of a number of pulse crops. We have cloned the bipartite genome of Mungbean Yellow Mosaic India Virus isolated from infected Soybean. We report here that agroinfection of Soybean seedlings with a single uncut recombinant binary plasmid containing tandem dimers of both DNA A and DNA B resulted in 100% infectivity in susceptible varieties. To understand the mechanism of natural resistance in a Soybean variety, we compared the abundance of the viral RNAs in a resistant and a susceptible variety at the early time points after agroinfection. Whilst the resistant variety displayed synthesis but rapid degradation of the early viral RNAs; the degradation in the susceptible variety was delayed resulting in accumulation of those transcripts later in infection. Accumulation of the late viral transcripts and DNA replication were detectable only in the susceptible variety. This indicates that rapid degradation of the early viral transcripts, possibly through siRNA mechanism, is one of the probable mechanisms of natural resistance against geminivirus.


Investigations into genotypic variations of peanut carbohydrates.
Carbohydrates are known to be important precursors in the development of roasted peanut quality. However, little is known about their genotypic variation. A better understanding of the role of carbohydrates in roasted peanut quality requires first an understanding of the genotypic variation in the soluble carbohydrate components. Ion exchange chromatography was used to isolate 20 different carbohydrate components in 52 genotypes grown in replicated trials at two locations. Inositol, glucose, fructose, sucrose, raffinose, and stachyose were quantitated, and 12 unknown peaks were evaluated on the basis of the peak height of the unknown relative to the cellobiose internal standard peak height. Peaks tentatively identified as verbascose and ajugose could not be properly integrated because of tailing. Of the 18 carbohydrates that were estimated, 9 exhibited significant variation between test environments, 5 among market types, 14 among genotypes within market types, and 11 exhibited some significant form of genotype x environment interaction. Genotypes accounted for 38-78% of the total variation for the known components, suggesting that broad-sense heritability for these components is high. The observed high genotypic variation in carbohydrate components is similar to the high genotypic variation observed for the sweetness attribute in roasted peanuts, which raises the question regarding possible interrelationships. The establishment of such interrelationships could be most beneficial to peanut breeding programs to ensure the maintenance of flavor quality in future peanut varieties.


Mechanism of inhibition of duck hepatitis B virus polymerase by (-)-beta-L-2',3'-dideoxy-3'-thiacytidine.
We have used the endogenous reverse transcriptase reaction of viral core particles from duck liver to elucidate the mechanism of inhibition of duck hepatitis B virus (DHBV) replication by the nucleoside analog (-)-beta-L-2',3'-dideoxy-3'-thiacytidine (3TC). As is the case in human immunodeficiency virus replication, 3TC-5'-triphosphate (3TC-TP) acts as a chain terminator for the DNA polymerase activities. The results of several different experiments support this conclusion, which explains the potent activity of 3TC against the hepadnaviruses. In isolated DHBV core particles, 3TC-TP inhibited the reverse transcriptase in a manner that resembled competitive inhibition with respect to dCTP. However, the kinetics of inhibition was not linear on a double-reciprocal plot for the highest concentrations of 3TC-TP and the lowest concentration of dCTP. This anomaly would be expected if binding to the nucleotide site was followed by DNA chain termination. Calculations that used only the linear part of the curve yielded a Ki of 0.78 +/- 0.10 microM 3TC-TP. The inhibition of core particles incubated in vitro with 3TC-TP was not reversed by removal of the free inhibitor. 3TC-TP inactivated the reverse transcriptase activity in a concentration-dependent manner. The Km of the chain termination reaction was calculated at 0.71 +/- 0.05 microM. Similar competitive kinetics and irreversible inhibition were also obtained on the endogenous DNA polymerase from viral particles from serum, suggesting that 3TC-TP also acts as a chain terminator of the DNA-directed DNA polymerase of DHBV replication.(ABSTRACT TRUNCATED AT 250 WORDS)


[Relationship between sepsis and 4G/5G polymorphism within the promoter region of plasminogen activator inhibitor-1 gene].
To study the relationship between sepsis and a 4G/5G polymorphism within the promoter region of plasminogen activator inhibitor-1 (PAI-1) gene in the Han nationality. Peripheral blood sample were collected from 89 patients with severe sepsis, 58 males and 31 females, aged 56, with a APHCHII score of 22.5 +/- 2.1 and a mortality of 51%, and 100 sex and age-matched postoperative patients without severe sepsis, all of the Han nationality. 4G/5G polymorphism of PAI-1 gene was determined using polymerase chain reaction (PCR) followed by restriction enzyme analysis. The carrier rate of 4G/4G genotype among the sepsis patients was 0.44, significantly higher than that among the controls (0.25, P < 0.01). The 4G allele frequency of the sepsis patients was 0.65, significantly higher than that of the controls (0.54, P < 0.05). The carrier rate of 4G/4G genotype among the non-survivors in the sepsis group was 0.54; significantly higher than that among the survivors (0.34, P < 0.05); and the 4G allele rate in the non-survivors was 70%, not significantly different from that of the survivors (59%, P > 0.05). PAI-1 4G/5G polymorphism is associated with the susceptibility and outcome of severe sepsis, 4G/4G genotype and 4G allele may be genetic risk factors of severe sepsis.


Thermal damage to chloroplast envelope membranes.
Nuclear magnetic resonance was used to detect thermal injury to chloroplasts in vivo. A lesion occurs in the chloroplast envelope membrane at temperatures between 53 degrees C and 57 degrees C, depending on species, leaf condition, and heating rate. The injury is associated with a sudden loss of water from the chloroplast.


Staphylococcus-induced proliferative glomerulonephritis and cerebral hemorrhage - fatal complications in a young female with postpartum cardiomyopathy and an implanted left ventricular assist device: a case report and review of the literature.
Background: The continuous-flow left ventricular assist device (CF-LVAD) is used to save the lives of patients in the final stage of congestive heart failure, replacing the pump function of the left ventricle. Although quality of life increases significantly, CF-LVAD-related complications might prove fatal, as in the case presented in this paper.Methods: A 20-year-old female, during her second pregnancy, presented with signs of heart failure. Emergency caesarean section was necessary to save the baby, but peripartum cardiomyopathy developed in the mother. The use of an implantable cardioverter-defibrillator (ICD) was necessary 5 years later. As the clinical progression was unfavorable under medical treatment, with the patient reaching INTERMACS (Interagency Registry for Mechanically Assisted Circulatory Support) Profile 1 (refractory cardiogenic shock), the treatment of choice was the implantation of a CF-LVAD.Results: After 3 years of follow-up (at the age of 28), the patient presented with a positive hemoculture for Staphylococcus aureus. Prolonged antibiotic therapy and attentive follow-up was prescribed. Although an effective antiplatelet and anticoagulant treatment was applied, and despite therapeutic values of prothrombin time and international normalized ratio (INR), the patient died as result of a fatal cerebral hemorrhage. The autopsy also revealed septic emboli, disseminated intravascular coagulation, and focal proliferative glomerulonephritis.Conclusions: Although the benefits of CF-LVAD are significant, bleeding episodes can be severe and LVAD-associated infection can trigger glomerular injury and increase mortality.


Application of Deep Learning Methods for Pedestrian Collision Detection Using Dashcam Videos.
The goal of this study is to clarify the usefulness of deep learning methods for pedestrian collision detection using dashcam videos for advanced automatic collision notification, focusing on pedestrians, as they make up the highest number of traffic fatalities in Japan. First, we created a dataset for deep learning from dashcam videos. A total of 78 dashcam videos of pedestrian-to-automobile accidents were collected from a video hosting website and from the Japan Automobile Research Institute (JARI). Individual frames were selected from the video data amounting to a total of 1,212 still images, which were added to our dataset with class and location information. This dataset was then divided to create training, validation, and test datasets. Next, deep learning was performed based on the training dataset to learn the features of pedestrian collision images, which are images that capture pedestrian behavior at the time of the collision. Pedestrian collision detection performance of the trained model was evaluated as the percentage of correct predictions of pedestrian collisions in image data according to varied test sets with different combinations of characteristics. Our results for the proposed method show high-precision collision detection for daytime, clear pedestrian wrap trajectory accident data, including accurate detection of pedestrian collision location information. However, nighttime, unclear accident data resulted in false detection or no detection. Reduction of exposure value and resolution was confirmed to reduce detection rate. The results of the present study suggest the possibility of pedestrian collision detection by deep learning using dashcam videos.


Human umbilical artery for microvascular grafting. Experimental study in the rat.
As microvascular surgery has developed, the necessity for smaller arterial prostheses of less than 2 mm in internal diameter (ID) has increased markedly. Glutaraldehyde-stabilized human umbilical arteries with 1.5 to 2.0 mm ID are smaller than any other graft material currently available. This study was designed to determine whether this graft material has potential clinical application for microanastomosis. Twenty male albino rats, each weighing 400 to 500 gm, were used in this study. The abdominal aorta was exposed and a 3- to 4-mm segment was resected. A 10-mm interposition graft of glutaraldehyde-stabilized human umbilical artery was implanted by end-to-end anastomosis. The patency of the grafts was determined by repeated operative exploration at intervals of 1 week, and 1, 3, 9, and 12 months. Two grafts were found to be occluded each time at the 1-, 3-, and 9-month explorations. At each exploration time, five rats were sacrificed and histopathological studies conducted. All five remaining grafts were patent at 12 months. There was no evidence of endothelial proliferation on the luminal surface of the patent grafts as determined by scanning electron microscopy. The luminal surface and underlying region consisted of an amorphous proteinaceous-like material. Significant degeneration of the vessel wall was noted in all grafts surviving over 9 months. Central necrosis surrounded by a chronic inflammatory process that extended to and included the adventitia was observed in the occluded grafts. While the patency demonstrated was good, the tissue changes noted in the walls of the grafts indicate that further study is necessary before this material can be used as a graft in humans.


Developing decision support systems for integrated coastal management in the tropics: is the ICM decision-making environment too complex for the development of a useable and useful DSS?
Integrated coastal management in the tropics requires the conservation of vulnerable and diverse ecosystems such as coral reefs and mangroves as well as the management of land and marine-based human activities. Decision-making for integrated coastal management involves multiple decision-makers and multiple stakeholders often with conflicting needs and interests. Decision support systems can be developed to improve our understanding of the inter-relationships between the natural and socio-economic variables and hence result in improved decision-making. The question is whether this decision making environment is actually too complex for the development of useful and useable decision support systems. This paper describes the components of the decision making environment and the components of a decision support system. It also explores the various techniques available to deal with different modelling needs, the constraints of inadequate data and the multi-objective decision making environment. In addition, different techniques of developing decision support systems can play important roles within integrated coastal management. Three coastal decision support systems are evaluated in terms of their design and role in integrated coastal management and are used to evaluate the potential to develop decision support systems for integrated coastal management.


Orientation and oligomerization specificity of the Bcr coiled-coil oligomerization domain.
The Bcr oligomerization domain, from the Bcr-Abl oncoprotein, is an attractive therapeutic target for treating leukemias because it is required for cellular transformation. The domain homodimerizes via an antiparallel coiled coil with an adjacent short, helical swap domain. Inspection of the coiled-coil sequence does not reveal obvious determinants of helix-orientation specificity, raising the possibility that the antiparallel orientation preference and/or the dimeric oligomerization state are due to interactions of the swap domains. To better understand how structural specificity is encoded in Bcr, coiled-coil constructs containing either an N- or C-terminal cysteine were synthesized without the swap domain. When cross-linked to adopt exclusively parallel or antiparallel orientations, these showed similar circular dichroism spectra. Both constructs formed coiled-coil dimers, but the antiparallel construct was approximately 16 degrees C more stable than the parallel to thermal denaturation. Equilibrium disulfide-exchange studies confirmed that the isolated coiled-coil homodimer shows a very strong preference for the antiparallel orientation. We conclude that the orientation and oligomerization preferences of Bcr are not caused by the presence of the swap domains, but rather are directly encoded in the coiled-coil sequence. We further explored possible determinants of structural specificity by mutating residues in the d position of the coiled-coil core. Some of the mutations caused a change in orientation specificity, and all of the mutations led to the formation of higher-order oligomers. In the absence of the swap domain, these residues play an important role in disfavoring alternate states and are especially important for encoding dimeric oligomerization specificity.


[Anaesthesia during COVID-19 epidemic].
Surgical pathway has gained in complexity thanks to the recent COVID-19 pandemic. The anesthetic management of the SARScoV-2 (+) patient imposes several modifications, and remains a work in progress. As a consequence there has been a shift to distant anaesthetic consultations while in-hospital consultations are more difficult to organize. Patients scheduled for surgery may benefit from a preoperative diagnostic testing for SARS coV2 infection if they have been in close contact with a COVID-19 patient, or if they are symptomatic or in case of major surgery. According to the PCR results, patients could have their surgery postponed, or may alternatively follow a specific clinical pathway in the operating and recovery rooms with an adapted anaesthetic plan.


Cholinesterases from plant tissues: I. Purification and characterization of a cholinesterase from mung bean roots.
A cholinesterase was purified 36-fold from mung bean (Phaseolus aureus) roots by a combination of differential extraction media and gel filtration. The enzyme could be effectively extracted only by high salt concentration, indicating that it is probably membrane-bound. Methods used for assaying animal cholinesterases were tested, two of which were adapted for use with the bean cholinesterase. The bean enzyme hydrolyzed choline and noncholine esters but showed its highest affinity for acetylcholine and acetylthiocholine. The pH optimum was 8.5 for acetylthiocholine and 8.7 for acetylcholine. The Michaelis constants were 72 and 84 mum for acetylcholine and acetylthiocholine, respectively. The cholinesterase was relatively insensitive to eserine (half-maximum inhibition at 0.42 mm) but showed high sensitivity to neostigmine (half-maximum inhibition at 0.6 mum). Other animal cholinesterase inhibitors were also found to inhibit the bean enzyme but most of them at higher concentrations than are generally encountered. Choline stimulated enzymatic activity. The molecular weight of the cholinesterase was estimated to be greater than 200,000, but at least one smaller form was observed. It is suggested that the large form of cholinesterase is converted to the smaller form by proteolysis.


Reprogramming in inter-species embryonal carcinoma-somatic cell hybrids induces expression of pluripotency and differentiation markers.
Somatic cell reprogramming holds great promise for the development of novel cellular therapeutics. A number of sources of reprogramming potential have been identified, including oocytes, embryonic germ (EG) cells and embryonic stem (ES) cells. However, each of these sources of reprogramming factors is problematic, since they are either not freely available or have special growth requirements. Embryonal carcinoma (EC) cells are another source of pluripotent cells that, unlike ES and EG cells, do not usually require special growth conditions. Since they share many of the key characteristics of ES cells, such as pluripotency, EC cells may provide a readily amenable alternative source of reprogramming factors and could serve as a model for ES cells in this respect. Here we show that mouse EC cells can also function as donors of reprogramming factors. PEG-mediated fusion between murine EC cells (P19) and the cells of a human T-lymphoma cell line (CEM-GFP) resulted in inter-species hybrid colony formation. Colonies of hybrid cells displayed heterogeneity in cellular morphology as well as in their pattern of human gene expression. Expression of two human transcription factors characteristic of undifferentiated pluripotent stem cells, Oct-4 and Sox-2, was detected in the hybrid cells, demonstrating activation of endogenous human markers of pluripotency. Simultaneously, down-regulation of CD45, a marker present in lymphocytic cells, was observed in some hybrids. The detection of human specific markers of differentiation, such as nestin, lamininbeta1, and collagen IValpha1, indicates that fusion resulted in reprogramming of the human cells to reflect the differentiation potential of the murine EC partner.


Evolution of seed dormancy due to sib competition: effect of dispersal and inbreeding.
The effect of dispersal and inbreeding on the evolution of seed dormancy to avoid sib competition is theoretically investigated, using a model which assumes a plant population with patchy spatial structure in a constant environment. Applying the inclusive fitness method, the evolutionarily stable dormancy rates are analytically derived for three cases: (a) an asexual haploid population, (b) a diploid-hermaphrodite population in which the dormancy rate is controlled by seeds, and (c) a diploid-hermaphrodite population in which the dormancy rate is controlled by mother plants. The evolutionarily stable dormancy rates decrease in the order of case (c), case (a), and case (b). In all the cases, the evolutionarily stable dormancy rates increase with decreasing the dispersal rate. Although inbreeding generally increases the evolutionarily stable dormancy rates, inbreeding due to selfing reduces the rate exceptionally in case (c).


Conservation of RET proto-oncogene splicing variants and implications for RET isoform function.
The RET proto-oncogene encodes a receptor tyrosine kinase required for development of the kidney and neural crest-derived cell types. Alternative splicing of the 3' exons of human RET results in three protein isoforms with distinct C-termini: RET9, RET51, and RET43. These RET isoforms show differential binding to downstream adapter molecules, suggesting they may have distinct signaling functions. We have characterized Ret 3' sequences in mouse and investigated alternative splicing of this region. We found that the organization of Ret 3' sequences is very similar to human RET. The mouse locus also has alternatively spliced C-terminal coding regions, and the sequences corresponding to RET9 and RET51 are highly conserved in both position and sequence with the human locus. Further, we compared the predicted C-terminal amino acids of RET9 and RET51 in seven vertebrate species, and found that they are well conserved. We have identified sequence encoding a putative ret43 isoform in mouse, however the predicted amino acid sequence showed low homology to human RET43. Our data suggest that RET isoforms are evolutionarily highly conserved over a broad range of species, which may indicate that each isoform has a distinct role in normal RET function.


Polyamine depletion attenuates isoproterenol-induced hypertrophy and endoplasmic reticulum stress in cardiomyocytes.
Polyamines (putrescine, spermidine and spermine) play an essential role in cell growth, differentiation and apoptosis. Hypertrophy is accompanied by an increase in polyamine synthesis and endoplasmic reticulum stress (ERS) in cardiomyocytes. The present study was undertaken to elucidate the molecular interactions between polyamines, ERS and cardiac hypertrophy. Myocardial hypertrophy was simulated by incubating cultured neonatal rat cardiomyocytes in 100 nM isoproterenol (ISO). Polyamine deletion was achieved using 0.5 mM difluoromethylornithine (DFMO). Hypertrophy was estimated using cell surface area measurements, total protein concentrations and atrial natriuretic peptide (ANP) gene expression. Apoptosis was measured using flow cytometry and transmission electron microscopy. Expression of ornithine decarboxylase (ODC) and spermidine/spermine N1-acetyltransferase (SSAT) were analyzed via real-time PCR and Western blotting. Protein expression of ERS and apoptosis factors were analyzed using Western blotting. DFMO (0.5 mM and 2 mM) treatments significantly attenuated hypertrophy and apoptosis induced by ISO in cardiomyocytes. DFMO also decreased lactate dehydrogenase (LDH) and malondialdehyde (MDA) level in the culture medium. In addition, DFMO (0.5 mM) down regulated the expression of ODC, glucose-regulated protein 78 (GRP78), C/EBP homologous protein (CHOP), cleaved caspase-12, and Bax and up regulated the expression of SSAT and Bcl-2. Finally, these changes were partly reversed by the addition of exogenous putrescine (0.5 mM). The data presented here suggest that polyamine depletion could inhibit cardiac hypertrophy and apoptosis, which is closely related to the ERS pathway.


Measurement of the adhesion force between carbon nanotubes and a silicon dioxide substrate.
Carbon nanotube adhesion force measurements were performed on single-walled nanotubes grown over lithographically defined trenches. An applied vertical force from an atomic force microscope (AFM), in force distance mode, caused the tubes to slip across the 250-nm-wide silicon dioxide trench tops at an axial tension of 8 nN. The nanotubes slipped at an axial tension of 10 nN after being selectively coated with a silicon dioxide layer.


Prednisone withdrawal in pediatric kidney transplant recipients on tacrolimus-based immunosuppression: four-year data.
Corticosteroids have been used in renal transplant immunosuppression for over 40 yr. Despite their adverse effects, steroid therapy continues to be part of early as well as maintenance immunosuppression in most pediatric renal transplant centers. The association of steroids with growth retardation, weight gain, and acne may be particularly distressing during the critical years of adolescence and young adulthood, increasing the risk of medication non-adherence. This study reviews the outcomes of pediatric renal transplant patients treated with low-dose tacrolimus, mycophenolate mofetil, or azathioprine, and planned prednisone withdrawal. Thirty-seven pediatric renal transplant recipients were withdrawn from steroids. The mean follow-up after steroid withdrawal was 42+/-19 months. Graft and patient survival were 100%. The mean serum creatinine levels and calculated creatinine clearances remained stable throughout the period of observation. The mean creatinine clearance was 96+/-24 mL/min/1.73 m2 at steroid withdrawal and 93+/-20 mL/min/1.73 m2 at the latest follow-up. Five patients restarted prednisone; in four (11%) it was for suspected or confirmed acute rejection. Improvements were observed in serum lipid profiles, blood pressure, and body mass index. Most patients experienced catchup or stable growth after prednisone withdrawal. Four patients developed viral infections; all were successfully treated. The potential benefits of steroid withdrawal in pediatric renal transplantation are supported by our results.


Rivaroxaban compared to no treatment in ER-negative stage I-III early breast cancer patients (the TIP Trial): study protocol for a phase II preoperative window-of-opportunity study design randomised controlled trial.
Breast cancer patients are at a four-fold increased risk of developing a venous thromboembolism (VTE), a major cause of death in this group. Conversely, coagulation factors promote tumour growth and metastasis. This has been evidenced in preclinical models, with an inhibitory effect of anticoagulants on cancer growth through proliferative, angiogenic, apoptotic, cancer stem cell and metastatic processes. The extrinsic clotting pathway is also more upregulated in patients in the relatively poorer prognosis oestrogen receptor (ER)-negative breast cancer subgroup, with increased tumour stromal expression of the coagulation factors Tissue Factor and thrombin. Rivaroxaban (Xarelto®, Bayer AG, Leverkusen, Germany) is a direct oral anticoagulant (DOAC). It is a Factor Xa inhibitor that is routinely prescribed for the prevention of stroke in non-valvular atrial fibrillation and for both VTE prophylaxis and treatment. This trial will assess the anti-proliferative and other anti-cancer progression mechanisms of Rivaroxaban in ER-negative early breast cancer patients. This UK-based preoperative window-of-opportunity phase II randomised control trial will randomise 88 treatment-naïve early breast cancer patients to receive 20 mg OD Rivaroxaban treatment for 11 to 17 days or no treatment. Treatment will be stopped 24 h (range 18-36 h) prior to surgery or repeat core biopsy. All patients will be followed up for 2 weeks following surgery or repeat core biopsy. The primary endpoint is change in tumour Ki67. Secondary outcome measures include tumour markers of apoptosis and angiogenesis, extrinsic clotting pathway activation and systemic markers of metastasis, tumour load and coagulation. Laboratory evidence supports an anti-cancer role for anticoagulants; however, this has failed to translate into survival benefit when trialled in patients with metastatic disease or poor prognosis cancers, such as lung cancer. Subgroup analysis supported a potential survival benefit in better prognosis advanced disease patients. This is the first study to investigate the anti-cancer effects of anticoagulants in early breast cancer. UK National Research Ethics Service (NRES) approval 15/NW/0406, MHRA Clinical Trials Authorisation 48380/0003/001-0001. The sponsor is Manchester University NHS Foundation Trust, and the trial is co-ordinated by Cancer Research UK Liverpool Cancer Trials Unit (LCTU). EudraCT 2014-004909-33 , registered 27 July 2015. ISRCTN14785273 .


Antibiotic susceptibility profile of Aeromonas spp. isolates from food in Abu Dhabi, United Arab Emirates.
A total of 57 Aeromonas isolates from food samples such as fresh and frozen chicken, game birds, pasteurized milk, baby food, bakery products, fruit and vegetables, fish, and water from Abu Dahbi, UAE were investigated for antibiotic susceptibility profile. Most strains were resistant to penicillins (ticarcillin, mezlocillin, oxacillin, piperacillin), sulfamethoxazole, trimethoprim and macrolides (erythromycin, vancomycin, clindamycin) but sensitive to tetracycline, chloramphenicol, nitrofurantoin, aminoglycosides (amikacin, gentamicin, tobramycin), cephalosporins (cefuroxime, ceftrioxone, cefazolin, cephalexin, cephalothin, cefoxitin, cefotaxime), quinolone (ciprofloxacin), colistin sulphate and SXT (trimethoprim-sulfamethoxazole). On the other hand, many antibiotics showed excellent inhibitory activity (>75% strains were sensitive to them) against all the strains tested. These include cefuroxime, ceftrioxone, ciprofloxacin, colistin, amikacin, gentamicin, tetracycline, chloramphenicol, nitrofurantoin, cefotaxime and tobramycin. In conclusion, the results show a detailed pattern of sensitivity of the various Aeromonas spp. isolates to a variety of antibiotics and provide useful information in the context of selective isolation and phenotypic identification of the aeromonads from food.


Acetaminophen-induced immunosuppression associated with hepatotoxicity in mice.
We investigated the influence of acetaminophen (APAP), an analgesic and hepatotoxic agent, on the immune system in mice. The activity of serum glutamic-pyruvic transaminase was markedly increased by about 200 fold compared to that of the vehicle control following intraperitoneal injection of 400 mg/kg of APAP. In vivo antibody-producing responses to SRBC was significantly inhibited by APAP in a dose-dependent manner, while in vivo T cell-independent antibody-producing responses to TNP-Ficoll was not inhibited. The addition of thymocytes from APAP-treated mice suppressed the response to SRBC in vitro. Thymocyte blastogenesis following mitogenic stimulation with concanavalin A was also inhibited by injection of APAP. The delayed-type hypersensitivity response and mixed lymphocyte reaction, which are used to evaluate cell-mediated immunity, were also significantly reduced after treatment with APAP. These results indicate that APAP suppresses the humoral and cell-mediated immune responses at a dose that causes liver injury.


Polyphenols and red wine as peroxynitrite scavengers: a chemiluminescent assay.
A novel chemiluminescent assay for evaluating peroxynitrite (ONOO(-))-scavenging capacity was developed. The experimental protocol ensures sensitivity and reproducibility of measurements. The addition of 0-500 microM ONOO(-) to rat liver homogenate generated a luminous signal that was analyzed by chemiluminescence in a LKB Wallac liquid scintillation counter. The obtained optimal conditions were: 1-2 mg/mL of homogenate protein in 120 mM KCl, 30 mM phosphate buffer (pH 7.4), and 220 microM ONOO(-) at 30 degrees C. As polyphenols we used (+)-catechin, (-)-epicatechin, and myricetin. The most efficient of the compounds tested was myricetin with an IC(50) of 20 microM. The effectiveness of this method was verified by evaluating the antioxidant ability of three red wine samples to decrease peroxynitrite-initiated chemiluminescence. The ONOO(-)-scavenging activity of wines measured by this assay was related to the phenolic level of the samples. The quickness and reliability of this assay makes it particularly suitable for a large-scale screening of watery food extracts.


Comparison of changes in pulse wave velocity in patients on continuous ambulatory peritoneal dialysis and hemodialysis one year after introduction of dialysis therapy.
Damage to large arteries is a major contributory factor to the high cardiovascular morbidity and mortality of patients with end-stage renal disease. Among the methods for assessment of arterial damage, measurements of pulse wave velocity (PWV) are considered to predict cardiovascular risk. In the present study, we compared changes in the PWV of patients on hemodialysis (HD) and on continuous ambulatory peritoneal dialysis (CAPD). Patients were eligible for entry into the study if they had been on HD or CAPD for at least 3 months, but for fewer than 5 months, and if they had had no clinical cardiovascular disease during the 6 months preceding study entry. We followed a total of 22 patients (average age: 57.9 +/- 9.8 years) on HD and 22 patients (average age: 59.7 +/- 11.4 years) on CAPD for 1 year. Baseline PWV was determined, and a second measurement was performed 1 year after the baseline PWV had been obtained. Systolic and diastolic blood pressures in the two groups were similar [144.8 +/- 18.4 mmHg over 83.2 +/- 8.7 mmHg (CAPD) vs. 147.4 +/- 18.8 mmHg over 87 +/- 12.3 mmHg (HD)]. In the patients on CAPD, urine volume was fairly constant at about 546.1 +/- 365 mL daily; however, urine volume for most of the patients on HD was less than 100 mL daily. Although no significant difference was observed in baseline PWV [1726.2 +/- 374 m/s (CAPD) vs. 1774.2 +/- 531.36 m/s (HD)], a significant difference in PWV was seen between the two groups at 1 year [1631.9 +/- 380.8 m/s (CAPD) vs. 1853.2 +/- 434.2 m/s (HD), p < 0.05]. With the exception of level of serum cholesterol [256 + 24 mg/dL (CAPD) vs. 187 + 25 mg/dL (HD), p < 0.05], other laboratory data were not significantly different between the two groups. Despite higher serum cholesterol in the patients on CAPD, PWV was faster in the patients on HD. There may be less arterial damage in patients on CAPD than in patients on HD.


A role for Old Yellow Enzyme in ergot alkaloid biosynthesis.
Ergot alkaloids, secondary metabolites produced by filamentous fungi, elicit a diverse array of pharmacological effects. The biosynthesis of this class of natural products has not been fully elucidated. Here we demonstrate that a homologue of Old Yellow Enzyme encoded in the Aspergillus fumigatus ergot gene cluster catalyzes reduction of the alpha,beta unsaturated alkene of chanoclavine-I aldehyde 3. This reduction, which yields dihydrochanoclavine aldehyde, facilitates an intramolecular reaction between a secondary amine and aldehyde to form the D ring of the ergot alkaloid structural framework.


Cultural, gender, and socioeconomic contexts in therapeutic and social policy work.
The contention of this paper is that the context of social and therapeutic problems is critical to their resolution, and that many of them stem from historical and structural injustice. It focuses on the contextual issues of cultural, gender, and socioeconomic equity as providing important insights into authentic notions of social inclusion and well-being, and encourages therapists, service providers, researchers, and policy makers to take responsibility to ensure that these injustices are addressed, and become part of the public discourse about the sources and solutions of endemic social problems. Critique and deconstruction of institutional power in our public, private, and voluntary services is encouraged in a manner that honors diversity and enables sensitive therapy, other forms of service delivery and policy making that genuinely reflect the range of cultural, gender, and socioeconomic experiences of citizens.


Low-Power All-Organic Electrophoretic Display Using Self-Assembled Charged Poly( t-butyl methacrylate) Microspheres in Isoparaffinic Fluid.
The increasing demands for display devices with low power consumption and outdoor readability have stimulated comprehensive research into full-color reflective displays that employ color-tunable photonic crystal technologies. Although the recently developed crystalline colloidal arrays (CCAs) of the charged microspheres have shown the outstanding color tunability, the practical application is limited because the use of highly polar liquid medium such as water is required to maintain the charges on the surface of microsphere, whereas it is not suitable for long-term use in an electric field. Herein, a self-assembled CCA from charged poly( t-butyl methacrylate) microspheres was successfully fabricated, which was stabilized by the charged inverse micelles of sodium di-2-ethylhexyl-sulfosuccinate in a nonpolar isoparaffinic fluid. A charged all-organic CCA was found to exhibit full-color tunability with a 1000-fold reduction in the power consumption (∼6 μW cm-2) under a direct current voltage bias of 4 V in comparison to that in an aqueous system, which is a promising feature for a low-power-consumption display device.


Reduced Reference Perceptual Quality Model With Application to Rate Control for Video-Based Point Cloud Compression.
In rate-distortion optimization, the encoder settings are determined by maximizing a reconstruction quality measure subject to a constraint on the bitrate. One of the main challenges of this approach is to define a quality measure that can be computed with low computational cost and which correlates well with the perceptual quality. While several quality measures that fulfil these two criteria have been developed for images and videos, no such one exists for point clouds. We address this limitation for the video-based point cloud compression (V-PCC) standard by proposing a linear perceptual quality model whose variables are the V-PCC geometry and color quantization step sizes and whose coefficients can easily be computed from two features extracted from the original point cloud. Subjective quality tests with 400 compressed point clouds show that the proposed model correlates well with the mean opinion score, outperforming state-of-the-art full reference objective measures in terms of Spearman rank-order and Pearson linear correlation coefficient. Moreover, we show that for the same target bitrate, rate-distortion optimization based on the proposed model offers higher perceptual quality than rate-distortion optimization based on exhaustive search with a point-to-point objective quality metric. Our datasets are publicly available at https://github.com/qdushl/Waterloo-Point-Cloud-Database-2.0.


Characterization of peripheral T-cell lymphomas in a single North American institution by the WHO classification.
All peripheral T-cell lymphomas (PTCLs) diagnosed at a single institution were evaluated to determine the unique clinical features and outcome of specific entities and test the predictive validity of the International Prognostic Index (IPI). Cases of PTCL seen at the British Columbia Cancer Agency between 1981 and 2000 were identified. Pathologic material was re-assessed and classified according to the WHO classification, and patients were staged and treated uniformly according to era-specific guidelines. In total, there were 199 patients with PTCL and the most common subtypes were peripheral T-cell lymphoma unspecified (PTCL-US) (59%), anaplastic large-cell lymphoma, systemic type (ALCL) (17%) and extranodal NK/T-cell lymphoma, nasal and nasal-type (NASAL) (9%). Most patients were treated with CHOP-type chemotherapy. Three distinct prognostic subgroups were notable on survival analysis: favorable (cutaneous ALCL), 5-year overall survival (OS) 78%; intermediate [PTCL, ALCL and angioimmunoblastic lymphoma (AILT)], 5-year OS 35-43%; unfavorable [NASAL and enteropathy-type T-cell lymphoma (ETTL)], 5-year OS 22-24%. Furthermore, in PTCL-US and ALCL clinical separation of patients into good risk (IPI 0,1) and poor risk (IPI > or =2) subsets was demonstrated. A large proportion of PTCL patients have poor risk disease and/or a histologically aggressive subtype with frequent relapse and unfavorable outcome. For these patients, treatment with CHOP chemotherapy is only minimally effective and new strategies need to be developed, an effort that will require a multi-institution international collaboration due to the rarity of most subtypes.


TMS intensity and focality correlation with coil orientation at three non-motor regions.
Objective. The aim of this study is to define the best coil orientations for transcranial magnetic stimulation (TMS) for three clinically relevant brain areas: pre-supplementary motor area (pre-SMA), inferior frontal gyrus (IFG), and posterior parietal cortex (PPC), by means of simulations in 12 realistic head models of the electric field (E-field).Methods. We computed the E-field generated by TMS in our three volumes of interest (VOI) that were delineated based on published atlases. We then analysed the maximum intensity and spatial focality for the normal and absolute components of the E-field considering different percentile thresholds. Lastly, we correlated these results with the different anatomical properties of our VOIs.Results. Overall, the spatial focality of the E-field for the three VOIs varied depending on the orientation of the coil. Further analysis showed that differences in individual brain anatomy were related to the amount of focality achieved. In general, a larger percentage of sulcus resulted in better spatial focality. Additionally, a higher normal E-field intensity was achieved when the coil axis was placed perpendicular to the predominant orientations of the gyri of each VOI. A positive correlation between spatial focality and E-field intensity was found for PPC and IFG but not for pre-SMA.Conclusions. For a rough approximation, better coil orientations can be based on the individual's specific brain morphology at the VOI. Moreover, TMS computational models should be employed to obtain better coil orientations in non-motor regions of interest.Significance. Finding better coil orientations in non-motor regions is a challenge in TMS and seeks to reduce interindividual variability. Our individualized TMS simulation pipeline leads to fewer inter-individual variability in the focality, likely enhancing the efficacy of the stimulation and reducing the risk of stimulating adjacent, non-targeted areas.


Tests of the simple model of Lin and Brandts for the folding kinetics of ribonuclease A.
L.-N. Lin and J.F. Brandts recently proposed a simple model for the folding kinetics of ribonuclease A in which folding intermediates are not detectable. We have tested the basic assumption of the simple model for the major unfolded species, which is produced by a slow isomerization (the "X in equilibrium Y reaction" according to Lin and Brandts) after unfolding. The simple model assumes that in refolding the slow Y----X reaction must occur before any folding can take place. We have measured the Y----X reaction during folding. Tyrosine-detected folding occurs before the Y----X reaction; the difference in rate between the Y----X reaction and folding monitored by tyrosine absorbance becomes large when the stabilizing salt 0.56 M (NH4)2SO4 is added. The simple model predicts that the kinetic properties of the X in equilibrium Y reaction in unfolded ribonuclease are the same as those of tyrosine-detected folding. We find, however, that the kinetics of the X in equilibrium Y reaction in unfolded ribonuclease are independent of urea concentration, whereas the rate of tyrosine-detected folding decreases almost 100-fold between 0.3 and 5 M urea, as reported by Lin and Brandts. We point out that the kinetic properties of the X in equilibrium Y reaction in unfolded ribonuclease are characteristic of proline isomerization.


The prevalence and incidence of systemic lupus erythematosus in Nottingham, UK, 1989-1990.
The incidence and prevalence of systemic lupus erythematosus (SLE) in a well defined area in the Midlands was determined, by case ascertainment using multiple sources, during the period 1.5.89 to 30.4.90. This first such study of SLE in the UK showed incidence rates of 1.5/100,000/year for males and 6.5/100,000/year for females. The highest incidence was seen in age groups 40-49 and 50-59 years, with rates for females of 10.5 and 18.4/100,000/year respectively. Prevalence rates were 3.7/100,000/year for males and 45.4/100,000/year for females: SLE was found to be more prevalent amongst Afro-Caribbean groups. The socioeconomic status of the SLE patients was similar to the local study population, using social class by occupation and disadvantage by geographical area as indicators. Marked overlap between different sources of retrieval suggests that ascertainment of cases was high.


Using an advanced radiation therapy technique for T4 squamous cell carcinoma of the face.
Patients with locally advanced skin cancer often present an uncommon and unique treatment challenge. Surgical resection and reconstruction with an acceptable cosmetic outcome is difficult for larger lesions with deep infiltration into subcutaneous tissues. Radiation therapy has been shown to be an effective treatment modality for advanced non-melanoma skin cancers, with cure rates ranging 50-100%. In this case report, we discuss the efficacy and outcome of treatment using an advanced radiation therapy technique to a large T4 squamous cell carcinoma of the face. The patient responded favorably to the advanced radiation treatment course, and achieved a clinical complete response to therapy. No further intervention was required. Advanced radiation therapy techniques offered treatment advantages that resulted in greater tumor dose escalation and minimizing of patient morbidity. Patients with advanced skin cancers of the head and neck should be considered for definitive radiation therapy using advanced treatment techniques. The use of definitive RT only for tumors deemed unresectable, or for inoperable patients at presentation deserves reconsideration. Further investigation is warranted.


Viral Vectors Applied for RNAi-Based Antiviral Therapy.
RNA interference (RNAi) provides the means for alternative antiviral therapy. Delivery of RNAi in the form of short interfering RNA (siRNA), short hairpin RNA (shRNA) and micro-RNA (miRNA) have demonstrated efficacy in gene silencing for therapeutic applications against viral diseases. Bioinformatics has played an important role in the design of efficient RNAi sequences targeting various pathogenic viruses. However, stability and delivery of RNAi molecules have presented serious obstacles for reaching therapeutic efficacy. For this reason, RNA modifications and formulation of nanoparticles have proven useful for non-viral delivery of RNAi molecules. On the other hand, utilization of viral vectors and particularly self-replicating RNA virus vectors can be considered as an attractive alternative. In this review, examples of antiviral therapy applying RNAi-based approaches in various animal models will be described. Due to the current coronavirus pandemic, a special emphasis will be dedicated to targeting Coronavirus Disease-19 (COVID-19).


Second twin: quality of survival if born by breech extraction following internal podalic version.
The intrapartum management of the vertex-breech and vertex-transverse twin gestation is controversial. The fall in perinatal mortality rate to a low level has resulted in this parameter failing to be an adequate gauge of the safety of breech extraction and the answer lies in the quality of survival of the infants. Fifty-one twin pairs, collected over 12 years at the Mercy Hospital for Women, Melbourne, occurred where twin 2 was born by breech extraction following internal inversion and the control (twin 1) did not have this procedure performed. In 8 pairs either a stillbirth or neonatal death occurred; in one pair childhood death due to an accident (fire) occurred; in 4 pairs the parents refused entrance to the study as they perceived both twins to be similar; in 2 sets the assessment was incomplete; 11 sets were untraceable leaving 25 sets fully assessed as children ranging in age from 2 to 12 years. Growth, and psychological scores were not significantly different between twins 1 and 2 but 2 children had cerebral palsy and both were born by breech extraction following internal version at 29.2 and 30.1 weeks' gestation, respectively. Because of small numbers the results failed to achieve statistical significance and this study was unable to answer the question regarding the safety of breech extraction following internal version but did show that the majority of infants so born do well.


CDKN2A Determines Mesothelioma Cell Fate to EZH2 Inhibition.
Malignant pleural mesothelioma is an aggressive cancer, heterogeneous in its presentation and behaviour. Despite an increasing knowledge about molecular markers and their diagnostic and prognostic value, they are not used as much as they might be for treatment allocation. It has been recently reported that mesothelioma cells that lack BAP1 (BRCA1 Associated Protein) are sensitive to inhibition of the EZH2 (Enhancer of Zeste Homolog 2) histone methyltransferase. Since we observed strong H3K27me3 (histone H3 lysine 27 trimetylation) immunoreactivity in BAP1 wild-type mesothelioma biopsies, we decided to characterize in vitro the response/resistance of BAP1 wild-type mesothelioma cells to the EZH2 selective inhibitor, EPZ-6438. Here we demonstrate that BAP1 wild-type mesothelioma cells were rendered sensitive to EPZ-6438 upon SIRT1 (Sirtuin 1) silencing/inhibition or when cultured as multicellular spheroids, in which SIRT1 expression was lower compared to cells grown in monolayers. Notably, treatment of spheroids with EPZ-6438 abolished H3K27me3 and induced the expression of CDKN2A (Cyclin-Dependent Kinase Inhibitor 2A), causing cell growth arrest. EPZ-6438 treatment also resulted in a rapid and sustained induction of the genes encoding HIF2α (Hypoxia Inducible Factor 2α), TG2 (Transglutaminase 2) and IL-6 (Interleukin 6). Loss of CDKN2 is a common event in mesothelioma. CDKN2A silencing in combination with EPZ-6438 treatment induced apoptotic death in mesothelioma spheroids. In a CDKN2A wild-type setting apoptosis was induced by combining EPZ-6438 with 1-155, a TG2 selective and irreversible inhibitor. In conclusion, our data suggests that the expression of CDKN2A predicts cell fate in response to EZH2 inhibition and could potentially stratify tumors likely to undergo apoptosis.


Low-molecular-weight contact allergens in p-tert-butylphenol-formaldehyde resin.
p-tert-Butylphenol-formaldehyde resin (PTBPFR) is a contact allergen that is included in most standard patch test series. This resin consists of a large number of substances most of which are unknown. The aim of this study was to investigate if allergens in PTBPFR are molecules mainly of low (MW < 250), medium, or high molecular weight (MW > 1,000); to isolate and identify some of the low molecular weight components of the resin; and for new substances to determine if these were allergens. Gel permeation chromatography, patch testing, high-pressure liquid chromatography, mass spectrometry, and nuclear magnetic resonance spectrometry were used. Patch test reactions indicated allergens in low-, medium-, and high-molecular-weights fractions. The strongest patch test reactivity was seen to a medium molecular weight fraction constituting only 6% wt/wt of the resin for 4 of the patients. Two patients reacted positively to low molecular weight fractions, and one of these patients reacted only to these fractions. The following substances were isolated: 4-tert-butyl-[1,2]benzoquinone, 4-tert-butyl-2,6-bis-hydroxymethyl-phenol, 4-tert-butylbenzene-1,2-diol, 4-tert-butyl-2-hydroxymethyl-phenol, 5-tert-butyl-2-hydroxy-3-hydroxymethyl-benzaldehyde, 4-tert-butyl-2-hydroxymethyl-6-methoxymethyl-phenol, and p-tert-butylphenol. Patients reacted positively to 5-tert-butyl-2-hydroxy-3-hydroxymethyl-benzaldehyde and 4-tert-butyl-2-hydroxymethyl-6-methoxymethyl-phenol but not to 4-tert-butyl-[1,2]benzoquinone. Two new allergens in PTBPFR were found: 5-tert-butyl-2-hydroxy-3-hydroxymethyl-benzaldehyde and 4-tert-butyl-2-hydroxymethyl-6-methoxymethyl-phenol.


Human dissection: an approach to interweaving the traditional and humanistic goals of medical education.
Anatomy remains one of the core courses of medical school, but the time devoted to it is decreasing. To accommodate the explosion of medical knowledge, educators search to streamline the curriculum. Because it is time-consuming, dissection comes under increased scrutiny. Even in the face of these pressures to reduce course hours, I would like to propose broadening, not reducing, the responsibilities of the anatomy instructor. Anatomy instructors can play a crucial role in helping medical schools meet the critical need to cultivate humanistic values, especially in the arena of end-of-life care. Anatomy can--and should--play an important role in a curriculum-wide effort to address this issue. Just as dissection remains an essential technique to teach three-dimensional concepts, the cadaver dissection lab is an ideal place to introduce concepts of humanistic care. The lab evokes the students' memories, speculations, and fears about serious illness in themselves, their families, and loved ones. Some programs address these reactions with supplemental activities, such as journaling, essay writing, and small group discussion. Valuable as these activities may be, anatomy instructors can achieve more by recognizing their role as a mentor, who can integrate humanistic values into traditional course objectives in a way that adds little time to the curriculum. The attitude of the instructor in ministering to the students' needs as they undertake the emotionally charged task of dissection can provide a model for how the students will respond, in turn, to the hopes and fears of their patients-and to their own reactions to dying. This approach will allow students to implement and practice humanistic values immediately, laying a foundation for their clinical training.


Erb's palsy contrasted with Klumpke's and total palsy: different mechanisms are involved.
The purpose of this study was to examine the available evidence regarding the nature of the forces that were involved in the production of a lower plexus palsy or a total (whole arm) palsy, as contrasted with the nature of the forces that resulted in an upper plexus palsy. This was a review of studies that dealt with specific mechanisms that were supplemented by reports of total palsy that were gleaned from the literature and case reports from the clinical and medical legal cases of the authors. Studies of the forces involved in brachial plexus injury in adults and from cadaver studies in infants who were stillborn or who died in the newborn period attest to the nature of the forces needed to damage the lower plexus. This evidence is reinforced by the case reports. Forces other than simple widening of the head-shoulder angle are necessary to disrupt the roots or cords of the lower brachial plexus. The position of the arm and direction of the forces that are applied determine the nature of the lesion.


JAK2 V617F mutation is uncommon in non-Hodgkin lymphomas.
Recently, the JAK2 V617F mutation has been reported in high proportions of chronic myeloproliferative disorders, including polycythemia vera. To see whether the JAK2 V617F is important in the pathogenesis of lymphoid malignancies, this study analysed the occurrence of the JAK2 V617F mutation in 117 non-Hodgkin lymphomas (NHLs) by a single strand conformation polymorphism assay. However, there was no JAK2 V617F mutation in the NHLs and the data suggest that the JAK2 V617F mutation may not play a role in the development of NHL.


An ADAM9 mutation in canine cone-rod dystrophy 3 establishes homology with human cone-rod dystrophy 9.
To identify the causative mutation in a canine cone-rod dystrophy (crd3) that segregates as an adult onset disorder in the Glen of Imaal Terrier breed of dog. Glen of Imaal Terriers were ascertained for crd3 phenotype by clinical ophthalmoscopic examination, and in selected cases by electroretinography. Blood samples from affected cases and non-affected controls were collected and used, after DNA extraction, to undertake a genome-wide association study using Affymetrix Version 2 Canine single nucleotide polymorphism chips and 250K Sty Assay protocol. Positional candidate gene analysis was undertaken for genes identified within the peak-association signal region. Retinal morphology of selected crd3-affected dogs was evaluated by light and electron microscopy. A peak association signal exceeding genome-wide significance was identified on canine chromosome 16. Evaluation of genes in this region suggested A Disintegrin And Metalloprotease domain, family member 9 (ADAM9), identified concurrently elsewhere as the cause of human cone-rod dystrophy 9 (CORD9), as a strong positional candidate for canine crd3. Sequence analysis identified a large genomic deletion (over 20 kb) that removed exons 15 and 16 from the ADAM9 transcript, introduced a premature stop, and would remove critical domains from the encoded protein. Light and electron microscopy established that, as in ADAM9 knockout mice, the primary lesion in crd3 appears to be a failure of the apical microvilli of the retinal pigment epithelium to appropriately invest photoreceptor outer segments. By electroretinography, retinal function appears normal in very young crd3-affected dogs, but by 15 months of age, cone dysfunction is present. Subsequently, both rod and cone function degenerate. Identification of this ADAM9 deletion in crd3-affected dogs establishes this canine disease as orthologous to CORD9 in humans, and offers opportunities for further characterization of the disease process, and potential for genetic therapeutic intervention.


A scoping review of inequities in access to organ transplant in the United States.
Organ transplant is the preferred treatment for end-stage organ disease, yet the majority of patients with end-stage organ disease are never placed on the transplant waiting list. Limited access to the transplant waiting list combined with the scarcity of the organ pool result in over 100,000 deaths annually in the United States. Patients face unique barriers to referral and acceptance for organ transplant based on social determinants of health, and patients from disenfranchised groups suffer from disproportionately lower rates of transplantation. Our objective was to review the literature describing disparities in access to organ transplantation based on social determinants of health to integrate the existing knowledge and guide future research. We conducted a scoping review of the literature reporting disparities in access to heart, lung, liver, pancreas and kidney transplantation based on social determinants of health (race, income, education, geography, insurance status, health literacy and engagement). Included studies were categorized based on steps along the transplant care continuum: referral for transplant, transplant evaluation and selection, living donor identification/evaluation, and waitlist outcomes. Our search generated 16,643 studies, of which 227 were included in our final review. Of these, 34 focused on disparities in referral for transplantation among patients with chronic organ disease, 82 on transplant selection processes, 50 on living donors, and 61 on waitlist management. In total, 15 studies involved the thoracic organs (heart, lung), 209 involved the abdominal organs (kidney, liver, pancreas), and three involved multiple organs. Racial and ethnic minorities, women, and patients in lower socioeconomic status groups were less likely to be referred, evaluated, and added to the waiting list for organ transplant. The quality of the data describing these disparities across the transplant literature was variable and overwhelmingly focused on kidney transplant. This review contextualizes the quality of the data, identifies seminal work by organ, and reports gaps in the literature where future research on disparities in organ transplantation should focus. Future work should investigate the association of social determinants of health with access to the organ transplant waiting list, with a focus on prospective analyses that assess interventions to improve health equity.


Dominant social status and contraceptive hormone treatment inhibit atherogenesis in premenopausal monkeys.
The stress of social subordination is associated with exacerbation of coronary artery atherosclerosis in premenopausal cynomolgus monkeys, possibly as a result of the ovarian dysfunction that reliably accompanies subordinate social status. The primary objective of the current study was to determine whether treatment with an oral contraceptive (OC) provides relative protection from development of atherosclerotic plaques, especially among animals made vulnerable to atherosclerosis by social subordination. In the present study, 193 adult female monkeys (Macaca fascicularis) were placed in social groups of 5 or 6 animals each. Half of the animals were then fed an atherogenic diet to which had been added a triphasic OC, while the remainder received only the atherogenic diet. At the end of 26 months, atherosclerosis was measured in an iliac artery biopsy taken from each monkey. The results demonstrated that among untreated animals subordinate individuals developed significantly more atherosclerosis than did their dominant counterparts (P < .01); however, OC treatment inhibited atherosclerosis in subordinate animals (P < .05) and eliminated the difference between dominant and subordinate animals that was observed in the untreated condition. Subordinate social status and OC treatment were both associated with reduced plasma concentrations of HDL cholesterol (P < .01 for both), and subordinate monkeys also had elevations in LDL cholesterol plus VLDL cholesterol (P < .01). Nonetheless, the interaction between social status and OC treatment remained significant even after covariance adjustment for variation in plasma lipids. Taken together, these results suggest that social subordination worsens, whereas OC treatment inhibits, atherosclerosis, and that these effects are independent of concomitant variability in plasma lipids.


Simultaneous Determination of Antibodies to Pertussis Toxin and Adenylate Cyclase Toxin Improves Serological Diagnosis of Pertussis.
Serological diagnosis of pertussis is mainly based on anti-pertussis toxin (PT) IgG antibodies. Since PT is included in all acellular vaccines (ACV), serological assays do not differentiate antibodies induced by ACVs and infection. Adenylate cyclase toxin (ACT) is not included in the ACVs, which makes it a promising candidate for pertussis serology with the specific aim of separating infection- and ACV-induced antibodies. A multiplex lateral flow test with PT and ACT antigens was developed to measure serum antibodies from pertussis-seropositive patients (n = 46), healthy controls (n = 102), and subjects who received a booster dose of ACV containing PT, filamentous hemagglutinin, and pertactin (n = 67) with paired sera collected before and one month after the vaccination. If the diagnosis was solely based on anti-PT antibodies, 98.5-44.8% specificity (before and after vaccination, respectively) and 78.2% sensitivity were achieved, whereas if ACT was used in combination with PT, the sensitivity of the assay increased to 91.3% without compromising specificity. No increase in the level of anti-ACT antibodies was found after vaccination. This exploratory study indicates that the use of ACT for serology would be beneficial in combination with a lower quantitative cutoff for anti-PT antibodies, and particularly in children and adolescents who frequently receive booster vaccinations.


Synthesis of tetrahydroisoquinoline alkaloids via anodic cyanation as the key step.
We report a new route to tetrahydroisoquinoline (THIQ) alkaloids involving the alkylation of alpha-aminonitrile 2 as a key step. The latter compound was prepared by anodic cyanation of the corresponding tertiary amine 1. Reductive decyanation of alpha-aminonitriles 6a-c proceeded diastereoselectively (up to 95% de) to deliver the C1-substituted alkaloids precursors 9a-c. The syntheses of (+/-)-carnegine, (+/-)-norlaudanosine, and (+/-)-O,O-dimethylcoclaurine have been achieved.


Acute aquatic toxicity of sulfur mustard and its degradation products to Daphnia magna.
Sulphur mustard (HD) was the most widely produced chemical warfare agent (CWA) in the history of chemical warfare (CW). Simultaneously, the loads of HD account as by far the largest fraction of the sea-dumped CW. Nowadays its presence in the marine ecosystems recognized as a serious threat for marine users and maritime industries. Although, during over a decade of research much has been done to assess the environmental threats linked with underwater chemical munitions. There are, however, essential gaps in scientific knowledge including scarce information about the aquatic toxicity thresholds of HD and its degradation products. Standardized biotests were performed according to the Organisation for Economic Co-operation and Development (OECD) Test No. 202: Daphnia sp. Acute Immobilisation Test guidelines. Obtained results provide a solid foundation for comparison and categorisation of threats of HD and its degradation products. With the D. magna LC50 aquatic acute toxicity threshold at as low as 224 ± 12 μg × L-1, 1,2,5-trithiepane is very toxic, being one of the most toxic CWA degradation products that have been investigated up to date. It exhibits stronger effects than 1,4,5-oxadithiepane and diluted HD that turn out to be toxic. In total, the toxicity of 7 compounds has been estimated. Whenever possible, toxicity thresholds were compared with previously existing data originating from different biotests and mathematical modelling.


Antioxidant enzymes in red blood cells and lymphocytes of ataxia-telangiectasia patients.
Toxic oxygen metabolites may contribute to the development of tissue damage, and play a role in the pathogenesis of malignancies, some acute and chronic pulmonary diseases, and in cell damage by radiomimetic agents, which can be seen in patients with ataxia-telangiectasia (A-T). Oxidative stress resulting from increased free radical production and/or defects in antioxidant defences is also involved in neurodegenerative disorders. Thus, oxidative stress could account for several aspects of the pleiotropic phenotype of A-T patients. The aim of this study was to determine the activities of the enzymes involved in cellular antioxidant metabolism in A-T patients to see if there is any defect which may result in constant oxidative stress. Superoxide dismutase (SOD) and catalase activities of erythrocytes, in contrast to lymphocytes, were found to be significantly higher in patients than in healthy controls. Our results may be another indication for the presence of constant oxidative stress in A-T patients as suggested previously.


Early developing embryos affect the gene expression patterns in the mouse oviduct.
Fertilization and development of mouse embryos occur in the ampullae of oviduct. We hypothesize that fetal-maternal communication exists in the preimplantation period, allowing optimal development of embryos. It is known that embryotrophic factors from oviduct affect the development of embryos. Although embryos affect their own transport in the oviduct, the mechanism of action is unknown. As a step toward understanding the action of embryos on oviductal physiology, we adopted suppression subtractive hybridization (SSH) to compare the gene expression in the mouse oviduct containing early embryos with that of oviduct containing oocytes. Ten to twelve 1-cell mouse embryos were transferred to one oviduct of a foster mother and similar number of oocytes were transferred to the contralateral oviduct. The animals were sacrificed after 48 h and their oviducts were excised for mRNA study. Using SSH, we screened out 250 putative positive clones from the subtracted embryo-containing oviduct library and 97 of them were screened positive by reverse dot-blot analysis. DNA sequence analysis identified genes that shared high homology with sequences in GenBank/EMBL database with unknown functions. Overall, 13 of the 90 high-quality sequences (14%) were homologous to 6 different genes previously described. Reverse Northern analysis confirmed that the expression of these genes were higher in the embryo-containing oviduct than in the oocyte-containing oviduct. About 12% of these clones (11/90) were novel. This article is the first to report identification of genes in the oviduct that are upregulated in the presence of embryos during the preimplantation period.


Factors causing acute delirium in critically ill adult patients: a systematic review.
Delirium, also known as acute confusional state, is a clinical syndrome that is typified by a disturbance of consciousness, attention, cognition and perception. It is estimated that the incidence of delirium varies between 28.3% to 80% in intensive care patients. Delirium is most likely to affect older patients above the age of 65 years. The primary focus of this systematic review was to synthesise the best available evidence to identify the factors which are associated with acute delirium in critically ill adult patients. A three-step search strategy was utilised in this review. An initial limited search of MEDLINE and CINAHL was undertaken. A search strategy was then developed using identified keywords and MeSH headings. Lastly, the reference lists of all identified studies were examined. All searches were limited to English language studies published between 1990 and 2010. The participants of this review were adult patients in a critical care setting who present with acute delirium in the intensive care unit.The review considered studies which examine the factors causing delirium in critically ill adult patients, such as fever, hypoxia and sepsis.This review considered studies that included the incidence of acute delirium as related to the individual factors as an outcome measure.This review considered any high quality quantitative papers that addressed factors causing acute delirium in intensive care unit patients. This review included data from cohort and case control studies. The Joanna Briggs Institute Critical Appraisal Checklist for Cohort / Case Control Studies was used to assess methodological quality. Data were extracted using a standardised Data Extraction Tool from the Joanna Briggs Institute. Due to the heterogeneous nature of the study methods, the findings of this systematic review are presented in a narrative summary. Twenty-four papers were included in this review. Some factors were common; however certain factors were exclusive to the type of setting. With regards to pharmacological factors, benzodiazepines were the most likely medications to be associated with delirium as compared to other drugs used in the intensive care unit. With regards to biomarkers, a variety were implicated in causing delirium, such as apolipoprotein 4 genotype, plasma tryptophan, interleukin-6, cortisol and C-reactive protein. Older age was also found to be a common risk factor. With regards to the medical and surgical intensive care units, relatively similar risk factors of older age and co-morbidity are significant. In the cardiac intensive care unit, a variety of risk factors were significant. Benzodiazepines have been singled out as the most likely drug to cause delirium. The biomarkers cortisol and hypoalbuminemia were also shown to be significant risk factors of delirium. Future research should use increased sample sizes, recruited from more than one hospital, assessing diverse factors. Future studies could examine the effects of specific biomarkers on delirium. By creating a regression model for delirium prediction, clinicians may be able to create a checklist to identify patients at risk of developing delirium. This checklist could assist in monitoring patients at higher risk for developing delirium, allowing measures to be implemented for preventing the incidence or reducing the severity of delirium. The use of benzodiazepines should also be avoided if possible, as they are seen to have a significant association with the incidence of delirium.


Unprecedented double nucleophilic addition of a hydride at a central carbon of an eta3-allyl ligand.
Treatment of eta(3)-allyl compound [Cp(2)Mo(eta(3)-C(3)H(5))](+)(1; Cp =eta(5)-C(5)H(5)) with MH (M = Li, Na) resulted in reduction of the allyl ligand to give propane. Deuterium-labeling studies were used to trace the origins and fates of the hydrogen atoms. The mechanism is discussed in light of the HSAB principle. The studies showed that the formation of propane can be explained by 1,2-hydrogen migration from the central to the terminal carbon of the allyl ligand, and the subsequent double nucleophilic addition of the hydride at the central carbon.


Trien conjugates, schiff bases, and ceruloplasmin concentrations as markers of the onset of the risk of autodermoplasty complications.
Background. Surgical interference which includes debridement and further renewal of skin cover by autodermoplasty (ADP) is the most effective method of burn wound treatment. Skin graft failure is a serious complication of ADP procedures leading to the opening of previously closed wounds, graft loss, and an increased wound surface owing to the donor zone. Numerous factors are involved that may affect the results of the ADP procedure. When planning the terms of surgical interference, specialists have to take into consideration criteria regarding the visual readiness of the wound and the patient's objective state. Aim. We studied the possibility of using concentrations of trien conjugates, Schiff bases, and ceruloplasmin in the plasma of patients with deep local wounds as markers of the risk of autoskin graft failure. Methods. Ninety patients with local wounds of different origin and age were studied. Results and conclusions. It was found out that the amounts of trien conjugates, Schiff bases, and ceruloplasmin in plasma before the operation helped to predict the risk of autoskin graft failure with a high degree of probability (up to 92%). We established optimum cut-off points for trien conjugates, Schiff bases, and ceruloplasmin as indicators of the results of autodermoplasty operations, i.e. 350 mg/l, 0.35 units of oxidation indices (UOI), and 0.05 UOI, respectively.


Blood pressure and cationic transport systems during combined calcium channel blocker and calcium administration in males.
A double-blind, placebo-controlled parallel study was conducted on the effects of a high daily oral supplementation of 1 g elemental calcium, given twice daily for 16 weeks, followed by additional administration of 2 mg b.i.d. lacidipine or placebo for 8 weeks, on blood pressure, intracellular cationic concentrations and transport systems, plasma total calcium, ionized calcium and calciotropic hormones in normal male subjects. No significant difference in the effect of lacidipine vs. placebo on standing or recumbent systolic or diastolic blood pressure or heart rate was found between calcium- and placebo-treated subjects. The difference in lacidipine effect on erythrocyte Mg2+ concentration and platelet membrane cholesterol content between the placebo and calcium groups was statistically significant. There was no additional effect between groups of lacidipine on erythrocyte and platelet intracellular Ca2+, Na+ and K+ concentrations or transport systems and on plasma Ca2+ levels and calciotropic hormones. Combined calcium and lacidipine administrations in normal male subjects did not induce antihypertensive effects different from those induced by either agent alone.


[The prevention of infections in child daycare centers: potential influential factors].
The Greater Montreal Public health departments have produced and distributed to daycare centres a manual on the prevention of infections in daycare personnel and trainees. In order to identify which factors can influence daycare management in applying the prevention measures suggested in the manual, a study was carried out based on the Precede/Proceed theoretical framework. A total of 134 (31.6%) daycare directors completed the questionnaire. Many were aware of the different recommendations to prevent infections. However, two of the recommendations were not followed by many: wearing gloves and verifying the vaccination status of their personnel, especially in private daycare centres. Participants had a positive attitude toward the various recommendations but mentioned several problems, specifically difficulty in obtaining the information about the vaccination status, lack of time and money, as well as having to deal with a poor physical organization of the work place. Among others things, the participants proposed that there be a greater involvement from schools, provincial daycare regulators, community clinics and public health departments.


[Numerical impairments in genes in breast cancer: a multiplex ligation-dependent probe amplification study].
To analyze breast cancer samples using the new technique multiplex ligation-dependent probe amplification (MLPA) assay. MATERIAL AND METHODS; Formalin-fixed paraffin-embedded breast carcinoma samples from 65 patients were examined. After manual microdissection, DNA was isolated using a commercial kit ("QIAGEN") and analyzed with SALSA MLPA KIT P078-B1 Breast Tumour ("MRC-Holland"). Capillary electrophoresis provided results. MLPA assay was successful in all examined samples. The amplification and deletion frequencies of the analyzed genes were in line with the literature data. The technique requires conventional work-related skills in a molecular genetic laboratory and, as a whole, presents no problems with its usage. The interpretation of results is devoid of subjective meaning due to exclusively their mathematical analysis. MLPA assay provides an insight into numerical impairments in the following genes: ERBB2, MYC, TRAF4, C11orf30 (EMSY), ADAM9, IKBKB, CCNE1, TOP2A, CDH1, CDC6, ESR1, CPD, EGFR, MTDH, CCND1, BIRC5, MED1, FGFR1, MAPT, PRDM14, and AURKA. MLPA is an easy-to-use and promising method for multiplex genetic analysis of tumor cells in breast cancer.


Early life modulators and predictors of adult synaptic plasticity.
Early life experience can induce long-lasting changes in brain and behaviour that are opposite in direction, such as enhancement or impairment in regulation of stress response, structural and functional integrity of the hippocampus, and learning and memory. To explore how multiple early life events jointly determine developmental outcome, we investigated the combined effects of neonatal trauma (anoxia on postnatal day 1, P1) and neonatal novelty exposure (P2-21) on adult social recognition memory (3 months of age) and synaptic plasticity in the CA1 of the rat hippocampus (4.5-8 months of age). While neonatal anoxia selectively reduced post-tetanic potentiation (PTP), neonatal novel exposure selectively increased long-term potentiation (LTP). No interaction between anoxia and novelty exposure was found on either PTP or LTP. These findings suggest that the two contrasting neonatal events have selective and distinct effects on two different forms of synaptic plasticity. At the level of behaviour, the effect of novelty exposure on LTP was associated with increased social memory, and the effect of anoxia on PTP was not accompanied by changes in social memory. Such a finding suggests a bias toward the involvement of LTP over PTP in social memory. Finally, we report a surprising finding that an early behavioural measure of emotional response to a novel environment obtained at 25 days of age can predict adult LTP measured several months later. Therefore, individual differences in emotional responses present during the juvenile stage may contribute to adult individual differences in cellular mechanisms that underlie learning and memory.


Finite-size scaling analysis of pseudocritical region in two-dimensional continuous-spin systems.
At low temperatures, the two-dimensional continuous-spin systems exhibit large correlation lengths. Some of them show the Berezinskii-Kosterlitz-Thouless-like transitions, and some others show pseudocritical behaviors for which correlation lengths are extremely large but finite. To distinguish pseudo and genuine critical behaviors, it is important to understand the nature of spin-spin correlations and topological defects at low temperatures in continuous-spin systems. In this paper, I develop a finite-size scaling analysis which is suitable for distinguishing the critical behavior and its applications to the two-dimensional XY, Heisenberg, and RP(2) models.


New approximations, and policy implications, from a delayed dynamic model of a fast pandemic.
We study an SEIQR (Susceptible-Exposed-Infectious-Quarantined-Recovered) model due to Young et al. (2019) for an infectious disease, with time delays for latency and an asymptomatic phase. For fast pandemics where nobody has prior immunity and everyone has immunity after recovery, the SEIQR model decouples into two nonlinear delay differential equations (DDEs) with five parameters. One parameter is set to unity by scaling time. The simple subcase of perfect quarantining and zero self-recovery before quarantine, with two free parameters, is examined first. The method of multiple scales yields a hyperbolic tangent solution; and a long-wave (short delay) approximation yields a first order ordinary differential equation (ODE). With imperfect quarantining and nonzero self-recovery, the long-wave approximation is a second order ODE. These three approximations each capture the full outbreak, from infinitesimal initiation to final saturation. Low-dimensional dynamics in the DDEs is demonstrated using a six state non-delayed reduced order model obtained by Galerkin projection. Numerical solutions from the reduced order model match the DDE over a range of parameter choices and initial conditions. Finally, stability analysis and numerics show how a well executed temporary phase of social distancing can reduce the total number of people affected. The reduction can be by as much as half for a weak pandemic, and is smaller but still substantial for stronger pandemics. An explicit formula for the greatest possible reduction is given.


A New Preoperative Upright Design for Dermatochalasis Correction.
Conventional assessment of the height of upper eyelid skin excision in dermatochalasis correction is performed with patient's eyes closed in supine position. It is not able to consider the effects of gravity on the upper eyelid, thus may lead to asymmetric postoperative appearance. The authors herein report a novel preoperative upright design (PUD) that can accurately determine the amount of skin excision with patients' eyes open in dermatochalasis correction. Patients with dermatochalasis underwent PUD during blepharoplasty were enrolled and were followed-up for 9 to 15 months. A total of 116 patients (mean age 55.1 ± 6.1 years, range 46-78 years) successfully underwent the surgery. Using the PUD, the vertical height of skin excision was 8.2 ± 2.4 mm (6-19 mm), the preoperative margin fold distance was -0.5 ± 1.0 mm (-4 - 1 mm), which improved to 2.1 ± 0.6 mm (1-3 mm, P < 0.05) at the last follow-up visit. A total of 107 of 116 patients (92.2%) were judged as "good" (natural double eyelid folds with symmetric margin fold distance), 9 patients (7.8%) were judged as "fair" (natural double eyelid folds with the differences of margin fold distance between fellow eyelids within 2 mm), and no one was judged as "poor" (unsmooth double eyelid folds or the differences of margin fold distance between fellow eyelids is more than 2 mm). Preoperative upright design is a simple and effective method to accurately determine the amount of skin excision in blepharoplasty, and help to achieve symmetric double eyelids.


Laparoscopic small bowel resection of metastatic pulmonary carcinosarcoma.
Carcinosarcoma of the lung is a rare malignancy consisting of both epithelial and mesenchymal components. Although metastatic disease is common, metastatic disease to the small intestine is rare and has been reported only once in the literature. An unusual case of pulmonary carcinosarcoma with metastasis to the small bowel is presented. At the time of laparoscopic exploration, a 4 x 4 cm mass was seen in the jejunum involving the adjacent mesentery. The affected small bowel and mesentery were resected laparoscopically and bowel continuity was restored through an entirely intracorporeal technique. Laparoscopic small bowel resection can be performed safely in the setting of malignant disease and imparts many of the benefits of minimally invasive surgery.


Correlation between antibutyrylcholinesterasic and antioxidant activities of three aqueous extracts from Tunisian Rhus pentaphyllum.
For centuries, plants have been used in traditional medicines and there has been recent interest in the chemopreventive properties of compounds derived from plants. In the present study, we investigated the antibutyrylcholinestrasic (anti-BuChE) and antioxidant (against some free radicals) activities of extracts from Rhus pentaphyllum. Aqueous extracts were prepared from powdered R. pentaphyllum roots, leaves and seeds and characterized for the presence of tannins, flavonoids and coumarins. Seeds aqueous extract contained the highest quantities of both flavonoids and tannins (21.12% and 17.45% respectively). In the same way, seeds extracts displayed remarkable inhibition against BuChE over 95%, at 100 μg/ml and with IC50 0.74 μg/ml. In addition, compared to leaves and roots extracts, seeds aqueous extract revealed relatively strong antiradical activity towards the ABTS.+ (IC50 = 0.25 μg/ml) and DPPH (IC50 = 2.71 μg/ml) free radicals and decreased significantly the reactive oxygen species such O2.- (IC50 = 2.9 μg/ml) formation evaluated by the non-enzymatic generating O2.- system (Nitroblue tetrazolium/riboflavine). These data suggest that the anti-BuChE activities mechanism of these extracts occurs through a free radical scavenging capacities.The present study indicates that extracts of Rhus pentaphyllum leaves, seeds and roots are a significant source of compounds, such as tannins, flavonoids and coumarins, with anti-BuChE and antioxidant activities, and thus may be useful for chemoprevention.


Structure of Plant Cell Walls : XII. Identification of Seven Differently Linked Glycosyl Residues Attached to O-4 of the 2,4-Linked l-Rhamnosyl Residues of Rhamnogalacturonan I.
Seven differently linked glycosyl residues have been found to be glycosidically linked to O-4 of the branched 2,4-linked l-rhamnosyl residues contained in the rhamnosyl and galacturonosyl backbone of the cell wall pectic polysaccharide rhamnogalacturonan I. These seven glycosyl residues are, therefore, the first residues of at least seven different side chains attached to the rhamnogalacturonan backbone. These first side chain glycosyl residues are 5-linked l-arabinofuranosyl and terminal 3-, 4-, 6-, 2,6-, and 3,6-linked d-galactopyranosyl residues. The existence of at least seven different side chains in rhamnogalacturonan I indicates that rhamnogalacturonan I is either an exceedingly complex polysaccharide or that rhamnogalacturonan I is a family of polysaccharides with similar or identical rhamnogalacturonan backbones substituted with different side chains.


Receptor-independent modulation of reconstituted Galpha(i) protein mediated by liposomes.
A cationic amphiphile, BC5 (N-pentadecylpiperidin-4-amine), was recently designed and tested for its ability to directly stimulate the activity of recombinant Galpha inhibitory subunits. However, amphiphilic drugs can self-associate and bind to plasma membranes, causing undesired side effects. In this contribution, we report on the incorporation of BC5 in 1,2-dipalmytoyl-sn-glycerophosphocoline (DPPC) liposomes and on the characterization of the mixed DPPC/BC5 systems at various lipid/drug mole ratios by means of dynamic light scattering, differential scanning calorimetry and fluorescence spectroscopy. The myristoylated Galpha(i) subunit (Galpha-mir) was reconstituted in 1,2-dimiristoyl-sn-glycerophosphocoline (DMPC) bilayers, as a mimic of the drug target. We compare several reconstitution procedures in liposomes and present for the first time a complete characterization of a Galpha subunit reconstitution in model membranes in terms of protein activity as a function of the reconstitution protocol. The incorporation of the drug in DPPC bilayers resulted in enhanced Gi-modulating efficiency (evaluated in terms of binding to GTPgammaS (guanosine-5'-(gamma-thio)-triphosphate)). A correlation of the physico-chemical features and binding activity of protein-containing membrane model is proposed.


Constitutive endocytosis and degradation of the pre-T cell receptor.
The pre-T cell receptor (TCR) signals constitutively in the absence of putative ligands on thymic stroma and signal transduction correlates with translocation of the pre-TCR into glycolipid-enriched microdomains (rafts) in the plasma membrane. Here, we show that the pre-TCR is constitutively routed to lysosomes after reaching the cell surface. The cell-autonomous down-regulation of the pre-TCR requires activation of the src-like kinase p56(lck), actin polymerization, and dynamin. Constitutive signaling and degradation represents a feature of the pre-TCR because the gammadeltaTCR expressed in the same cell line does not exhibit these features. This is also evident by the observation that the protein adaptor/ubiquitin ligase c-Cbl is phosphorylated and selectively translocated into rafts in pre-TCR- but not gammadeltaTCR-expressing cells. A role of c-Cbl-mediated ubiquitination in pre-TCR degradation is supported by the reduction of degradation through pharmacological inhibition of the proteasome and through a dominant-negative c-Cbl ubiquitin ligase as well as by increased pre-TCR surface expression on immature thymocytes in c-Cbl-deficient mice. The pre-TCR internalization contributes significantly to the low surface level of the receptor on developing T cells, and may in fact be a requirement for optimal pre-TCR function.


The influence of macromolecular crowding on HIV-1 protease internal dynamics.
High macromolecular concentrations, or crowded conditions, have been shown to affect a wide variety of molecular processes, including diffusion, association and dissociation, and protein folding and stability. Here, we model the effect of macromolecular crowding on the internal dynamics of a protein, HIV-1 protease, using Brownian dynamics simulations. HIV-1 protease possesses a pair of flaps which are postulated to open in the early stages of its catalytic mechanism. Compared to low concentrations, close-packed concentrations of repulsive crowding agents are found to significantly reduce the fraction of time that the protease flaps are open. Macromolecular crowding is likely to have a major effect on in vivo enzyme activity, and may play an important regulatory role in the viral life cycle.


Harnessing the power of social media to support medical training during COVID-19.
Twitter offers a powerful means to share information, suggest ways to help and highlight useful initiatives during the global COVID-19 pandemic. We describe one successful Twitter campaign focusing on the role of medical students (#MedStudentCovid), led by the volunteer organisation Becoming A Doctor with support from leaders at the General Medical Council, Health Education England, NHS England and the World Health Organization.


Possible errors in pin insertion positions using the C-arm.
In order to measure errors in pin positions, a tibia with referenced pin insertions was radiographed in various locations using a mini C-arm. Changes in pin position locations up to seven millimeters and twelve degrees were found. To minimize these errors, limbs should be centered and parallel to the plane of the imaging screen of the C-arm. If possible, pin insertions should be made in this plane. Magnification due to the distance from the screen should be determined for precise pin positioning.


Functional Outcomes of Antegrade Homodigital Neurovascular Island Flaps for Fingertip Amputation.
Fingertip amputations can be debilitating and challenging to treat, due to the need for both function and cosmesis. We studied the functional outcomes of patients receiving an antegrade homodigital neurovascular island flap for fingertip amputations. From 2004 to 2013, we performed 13 cases of antegrade homodigital neurovascular island flaps. Functional outcomes were studied. Four had Allen Zone II and nine had Zone III injuries. Seven were performed as secondary surgery. The mean followup period was 36 weeks. All the flaps survived. Ten of 11 had less than or equal to 6 mm two-point discrimination. Five patients were noted to have mild flexion contracture at distal interphalangeal joints ranging from 5 to 10 degrees. There were three cases with nail deformity, three with fingertip hypersensitivity and four with cold intolerance. For patients injured on duty, the mean duration of sick leave taken was 18.5 weeks. With careful patient selection, the antegrade homodigital neurovascular island flap is a reliable means of restoring the function of the fingertip after amputation injury with significant tissue loss. Flap survival, sensibility and mobile adjacent joint can be expected, even when performed as a secondary surgery and for patients with work-related injuries.


Quantitative trait locus mapping of yield-related components and oligogenic control of the cap color of the button mushroom, Agaricus bisporus.
As in other crops, yield is an important trait to be selected for in edible mushrooms, but its inheritance is poorly understood. Therefore, we have investigated the complex genetic architecture of yield-related traits in Agaricus bisporus through the mapping of quantitative trait loci (QTL), using second-generation hybrid progeny derived from a cross between a wild strain and a commercial cultivar. Yield, average weight per mushroom, number of fruiting bodies per m(2), earliness, and cap color were evaluated in two independent experiments. A total of 23 QTL were detected for 7 yield-related traits. These QTL together explained between 21% (two-flushes yield) and 59% (earliness) of the phenotypic variation. Fifteen QTL (65%) were consistent between the two experiments. Four regions underlying significant QTL controlling yield, average weight, and number were detected on linkage groups II, III, IV, and X, suggesting a pleiotropic effect or tight linkage. Up to six QTL were identified for earliness. The PPC1 locus, together with two additional genomic regions, explained up to 90% of the phenotypic variation of the cap color. Alleles from the wild parent showed beneficial effects for some yield traits, suggesting that the wild germ plasm is a valuable source of variation for several agronomic traits. Our results constitute a key step toward marker-assisted selection and provide a solid foundation to go further into the biological mechanisms controlling productive traits in the button mushroom.


A synthetic peptide corresponding to the Rab4 hypervariable carboxyl-terminal domain inhibits insulin action on glucose transport in rat adipocytes.
The present study was conducted to examine the involvement of Rab4, a low molecular weight GTP-binding protein, in the action of insulin on glucose transport. A synthetic peptide corresponding to the Rab4 hypervariable carboxyl-terminal domain, Rab4-(191-210), was successfully transferred into rat adipocytes by electroporation and inhibited insulin-stimulated glucose transport by about 50% without affecting the basal transport activity. In contrast, synthetic peptides corresponding to the Rab3C and Rab3D carboxyl-terminal hypervariable domain had little effect on insulin action on glucose transport. The Rab4-(191-210) peptide also reduced insulin-induced GLUT4 translocation from the intracellular pool to the plasma membrane. Furthermore, the Rab4-(191-210) peptide reduced both insulin-induced glucose transport and GLUT4 translocation in the presence of a major histocompatibility complex class I antigen-derived peptide, D(k)-(62-85), which is a potent inhibitor of GLUT4 internalization, suggesting that the peptide inhibited exocytotic recruitment of GLUT4-containing vesicles. The Rab4-(191-210) peptide also inhibited GTP gamma S-stimulated glucose transport. In addition, insulin-stimulated glucose transport was inhibited by the addition of anti-Rab4 antibody. These results suggest that Rab4 protein plays a crucial role in insulin action on GLUT4 translocation, especially in exocytotic recruitment by the hormone of the glucose transporter to the plasma membrane from the intracellular retention pool.


Phase tipping: how cyclic ecosystems respond to contemporary climate.
We identify the phase of a cycle as a new critical factor for tipping points (critical transitions) in cyclic systems subject to time-varying external conditions. As an example, we consider how contemporary climate variability induces tipping from a predator-prey cycle to extinction in two paradigmatic predator-prey models with an Allee effect. Our analysis of these examples uncovers a counterintuitive behaviour, which we call phase tipping or P-tipping, where tipping to extinction occurs only from certain phases of the cycle. To explain this behaviour, we combine global dynamics with set theory and introduce the concept of partial basin instability for attracting limit cycles. This concept provides a general framework to analyse and identify easily testable criteria for the occurrence of phase tipping in externally forced systems, and can be extended to more complicated attractors.