Stabilization of model membranes during drying by compatible solutes involved in the stress tolerance of plants and microorganisms.
Many organisms accumulate compatible solutes under environmental stress conditions. Cyanobacteria accumulate compatible solutes in response to increased external salinity, with tolerance increasing from Suc (sucrose) or trehalose to 2-O-(alpha-D-glucopyranosyl)-glycerol and glycinebetaine accumulating species. It is not clear how these different solutes influence salt tolerance. One possible explanation may be a differential ability of these solutes to stabilize membranes under stress conditions. We therefore performed drying experiments with liposomes in the presence of compatible solutes. Suc, trehalose and sorbitol protected liposomes from leakage of a soluble marker and from membrane fusion during drying and rehydration. 2-O-(alpha-D-glucopyranosyl)-glycerol was less effective and glycinebetaine showed hardly any effect. In combination with Suc, the latter two solutes showed improved protection. Lipid-phase transitions are known to contribute to solute leakage from liposomes. We determined phase transitions in dry membranes in the absence or presence of the solutes, using Fourier-transform infrared spectroscopy. The ability of the solutes to decrease the phase transition temperature corresponded closely to their ability to protect the liposomes against solute leakage. All solutes interacted with the phosphate in the lipid headgroups. The magnitude of the shift in the asymmetric P=O stretching vibration correlated closely with the lipid-phase transition temperature. This indicates that the degree of membrane protection afforded by the solutes is mainly determined by their ability to interact with the membrane lipids. However, this is not a determinant of cellular protection against salt stress, as the solutes show a reverse order when ranked with regard to protection against these stresses.


Tropical eosinophilia. A human model of parasitic immunopathology, with observations on serum IgE levels before and after treatment.
The diverse clinical syndromes characterized by asthmatic symptoms, transient pulmonary infiltrates, and eosinophilia have tended to obscure the specific association of one such entity with filarial infections. Serum IgE levels were determined before and after therapy in a group of well-characterized patients with tropical eosinophilia (TE), studied earlier in Singapore. The mean serum IgE level in 14 cases before treatment with diethylcarbamazine was 2,355 ng. per milliliter, with a trend but statistically nonsignificant decrease in levels to 600-1,000 ng. occurring 8 to 12 weeks after therapy. Leukocyte and eosinophil counts showed a rapid reduction after treatment, and although mean complement-fixing (cf) titers to Dirofilarial antigen tended to decrease, they were not significantly reduced until 5 to 6 weeks. The historical development of evidence supporting the filarial etiology of TE was reviewed. Many basic questions engendered by the clinical syndrome of tropical eosinophilia make it an excellent model for study of the immunopathology of parasitic infections.


Hoof Disorders and Farriery in the Donkey.
This article provides a review of hoof anatomy and care in donkeys and mules. Hoof disease is a major cause of poor welfare and mortality globally. Problems associated with hoof disease are discussed in the context of behavior, diet, treatment, and prevention. The most common conditions encountered are discussed, including laminitis, the overgrown unbalanced hoof, white line disease, flexural deformities, and other significant issues. Differences between donkey and horse hoof anatomy are described.


High spatial-resolution imaging of label-free in vivo protein aggregates by VISTA.
Amyloid aggregation, formed by aberrant proteins, is a pathological hallmark for neurodegenerative diseases, including Alzheimer's disease and Huntington's disease. High-resolution holistic mapping of the fine structures from these aggregates should facilitate our understanding of their pathological roles. Here, we achieved label-free high-resolution imaging of the polyQ and the amyloid-beta (Aβ) aggregates in cells and tissues utilizing a sample-expansion stimulated Raman strategy. We further focused on characterizing the Aβ plaques in 5XFAD mouse brain tissues. 3D volumetric imaging enabled visualization of the whole plaques, resolving both the fine protein filaments and the surrounding components. Coupling our expanded label-free Raman imaging with machine learning, we obtained specific segmentation of aggregate cores, peripheral filaments together with cell nuclei and blood vessels by pre-trained convolutional neural network models. Combining with 2-channel fluorescence imaging, we achieved a 6-color holistic view of the same sample. This ability for precise and multiplex high-resolution imaging of the protein aggregates and their micro-environment without the requirement of labeling would open new biomedical applications.


Coral kin aggregations exhibit mixed allogeneic reactions and enhanced fitness during early ontogeny.
Aggregated settlement of kin larvae in sessile marine invertebrates may result in a complex array of compatible and incompatible allogeneic responses within each assemblage. Each such aggregate can, therefore, be considered as a distinct self-organizing biological entity representing adaptations that have evolved to maximize the potential benefits of gregarious settlement. However, only sparse information exists on the selective forces and ecological consequences of allogeneic coalescence. We studied the consequences of aggregated settlement of kin larvae of Stylophora pistillata (a Red Sea stony coral), under controlled laboratory settings. When spat came into contact, they either fused, establishing a chimera, or rejected one another. A one-year study on growth and survivorship of 544 settled S. pistillata genotypes revealed six types of biological entities: (1) Single genotypes (SG); (2) Bi-chimeras (BC); (3) Bi-rejecting genotypes (BR); (4) Tri-chimera entities (TC); (5) Three-rejecting genotypes (TR); and (6) Multi-partner entities (MP; consisting of 7.5 +/- 2.6 partners). Analysis of allorecognition responses revealed an array of effector mechanisms: real tissue fusions, transitory fusions and six other histoincompatible reactions (borderline formation, sutures, overgrowth, bleaching, rejection, and partner death), disclosing unalike onsets of ontogeny and complex modes of appearance within each aggregate. Evaluations at the entity level revealed that MP entities were the largest, especially in the first two months (compared with SG: 571% in the first month and 162% in the seventh month). However, at the genotype level, the SG entities were the largest and the colonies with the highest-cost-per-genotype were the TR and the MP colonies. The cost was calculated as reduced average genotype size, from 27% and 12% in the first month to 67% and 64% in the seventh month, respectively. In general, MP exhibited the highest survivorship rate (85%, after one year) and SG the lowest (54%). In view of the above, we suggest that the driving force behind gregarious kin settlements in Stylophora pistillata stems from gained benefits associated with the immediate and long-term increase in total size of the MP entity, whereas survivorship rates did not draw a parallel link. Furthermore, the biological organization of MP entity exhibits, simultaneously, an intricate network of rejecting and fusible interactions in a single allogeneic intimate arena, where proposed benefits surpass costs incurred by discord among founders. Above results and documentations on gregarious settlement in other marine taxa bring us to suggest that the 'group level' of kin aggregates may serve as a ubiquitous legitimate selection entity in the evolution of a sessile mode of life in marine organisms.


A Bottom-Up Approach for Developing Aptasensors for Abused Drugs: Biosensors in Forensics.
Aptamer-based point-of-care (POC) diagnostics platforms may be of substantial benefit in forensic analysis as they provide rapid, sensitive, user-friendly, and selective analysis tools for detection. Aptasensors have not yet been adapted commercially. However, the significance of the applications of aptasensors in the literature exceeded their potential. Herein, in this review, a bottom-up approach is followed to describe the aptasensor development and application procedure, starting from the synthesis of the corresponding aptamer sequence for the selected analyte to creating a smart surface for the sensitive detection of the molecule of interest. Optical and electrochemical biosensing platforms, which are designed with aptamers as recognition molecules, detecting abused drugs are critically reviewed, and existing and possible applications of different designs are discussed. Several potential disciplines in which aptamer-based biosensing technology can be of greatest value, including forensic drug analysis and biological evidence, are then highlighted to encourage researchers to focus on developing aptasensors in these specific areas.


Are 90% of deaths from cancer caused by metastases?
Numerous publications have stated that metastases are responsible for 90% of cancer deaths, but data underlying this assertion has been lacking. Our objective was to determine what proportions of cancer deaths are caused by metastases. Population-based data from the Cancer Registry of Norway for the years 2005-2015 was analyzed. We compared all deaths in the Norwegian population where a cancer diagnosis was registered as cause of death. Deaths caused by cancer, with and without metastases, were analyzed, by sex and tumor group. For solid tumors, 66.7% of cancer deaths were registered with metastases as a contributing cause. Proportions varied substantially between tumor groups. Our data support the idea that the majority of deaths from solid tumors are caused by metastases. Thus, a better understanding of the biology of metastases and identification of druggable targets involved in growth at the metastatic site is a promising strategy to reduce cancer mortality.


An integrated approach in a municipal WWTP: anaerobic codigestion of sludge with organic waste and nutrient removal from supernatant.
Co-digestion appears to be an interesting solution to increase the biogas production of poorly performing under-loaded digesters of waste activated sludge. In the Florence WWTP anaerobic codigestion could increase nitrogen and phosphorus loading rates and thus lower the nutrient removal efficiency. In order to develop an integrated solution to upgrade the Florence WWTP, the different process units were tested in experimental plants. Anaerobic codigestion with source-collected organic solid waste in a pilot-scale bioreactor showed an increase of GPR from 0.15 to 0.45 Nl biogas/l/d with 23% of organic waste loaded. Autotrophic nitrogen removal was carried out in two lab-scale pilot plants which were fed with a real anaerobic supernatant after phosphate removal via struvite formation. The nitritation MBBR has been working for one year at steady-state conditions with a perfect nitrite/ammonium ratio equal to 1:1. Anammox biomass enrichment was performed in a suspended biomass SBR and the specific nitrogen removal rate increased from 1.7 to 58 gN/kgVSS/d in 375 days.


Comparison of effects of bleach and cleansing tablet on reflectance and surface changes of a dental alloy used for removable partial dentures.
Proper hygienic care of removable partial dentures is important for maintaining a healthy oral mucosa. A daily home care routine is the responsibility of the patient to maintain oral hygiene. A dentist must inform the patient about denture cleansers used for plaque control. Common denture cleansers may negatively affect the surface of alloys used to make partial dentures. The purpose of this study was to evaluate whether bleach and cleansing tablet have deleterious effects on the surface of a dental alloy used to fabricate removable partial dentures. Nineteen test specimens were prepared from CoCr dental alloy. The specimens were cast into 10 mm long x 10 mm wide x 1 mm thick. Fresh solutions made from a commercial bleach and a commercial cleansing tablet were used for each treating. Three groups (commercial denture cleansing tablet, bleach and tap water) were tested. Six specimens were immersed in 200 mL of tap water containing a single denture cleansing tablet. Two spoons of the commercial bleach was added to 200 mL of tap water. Seven specimens were treated. The control specimens were immersed in 200 mL of tap water. Six specimens were the control group. Each specimen was immersed for 30 min per day in the solution for a period of 30 days. The specimens were analyzed for reflectance with a spectrophotometer, for surface changes with a light microscope and weight which was measured with sensitive balance before and after treatment. The SPSS program was used for the statistical analysis. The Kruskal-Wallis Test and Wilcoxon Signed Ranks Test was used. No significant changes of reflectance was shown in the Kruskal-Wallis Test (p > 0.05). The Wilcoxon Signed Ranks Test was performed for comparison into groups. The difference between before and after treatment was statistically significant for the commercial denture cleansing tablet and the commercial bleach (p = 0.028) (p < 0.05). Reflectance was decreased after treatment. Within the limitations of this study, the commercial cleaning solutions had influenced the decrease of reflectance on the surface of the dental alloy. This solutions should not be limitlessly used on dentures with metal components, because they caused clinically significant reflectance changes on the surface of the alloy after 30 days.


Navigating Food and Drug Administration Mandated Breast Implant Patient Literature: A Call to Increase Equitable Healthcare Access Within Plastic Surgery.
In October 2021, the US Food and Drug Administration mandated patient decision checklists and new labeling for breast implants with the goal of improving the informed decision-making process for patients considering breast implantation. Given growing concerns over breast implant-associated anaplastic large cell lymphoma, breast implant-associated squamous cell carcinoma, and breast implant illness, patients should be able to easily review these resources to make a fully informed decision when considering surgery. This study seeks to elucidate the accessibility, and therefore the utility of the newly mandated literature for the average breast implant patient. Patient decision checklists and breast implant boxed warnings were obtained from the most used breast implant manufacturers in the United States-Allergan, Mentor, and Sientra. Readability analysis of all Food and Drug Administration required documents was performed using the Flesch Reading Ease Score, Flesch Kincaid Grade Level, Gunning-Fog Index, Coleman-Liau Index, Simplified Measure of Gobbledygook, and Automated Readability Index. The overall readability of all Allergan, Mentor, and Sientra patient materials correlates with a college reading level. Documents from all 3 implant manufacturers were of a statistically significantly higher reading level than that recommended by the American Medical Association and US Department of Health and Human Services. No materials were found to be at or below the recommended sixth-grade level. The newly mandated breast implant patient decision guides are written at a college reading level. These materials should be simplified to improve health literacy shared decision making.


Bilateral pneumothorax after orthognatic surgery.
Among complications in orthognathic surgery, the insurgence of pneumothorax is very rare. Pneumothorax is the presence of air or gas in the pleural cavity and it is rare complications in the postoperative oral and maxillofacial surgery patient. The clinical results are dependent on the degree of collapse of the lung on the affected side. Pneumothorax can impair oxygenation and/or ventilation. If the pneumothorax is significant, it can cause a shift of the mediastinum and compromise haemodynamic stability. While 10% of pneumothoraces are asymptomatic, patients often complain of acute chest pain and difficulty breathing. There is a reduction in vital capacity, tachycardia, tachypnoea and a decrease in partial pressure of oxygen with an inability to maintain oxygen saturations. We observed this unusual surgical consequence in a 28-year-old female with negative clinical history and instrumental evaluation after Le Fort I osteotomy and bilateral sagittal split osteotomy (BSSO). No further consequences, no neurological sequelae, no infections and no other osteotomies sequelae were seen. Sudden post-surgical dispnea associated to sub-cutaneous emphysema of the neck and of the thorax must be adequately observed with the aim of monitoring further severe sequelae. The anaesthetic management of the emergency difficult airway in any post-surgical orthognatic treatment can be extremely difficult requiring a multi-disciplinary approach.


In vitro evaluation of the synergistic antiviral effects of acemannan in combination with azidothymidine and acyclovir.
The antiviral effects of selected combinations between acemannan (ACE-M), a long-chained, polydispersed, beta-(1,4)-acetylated mannan, were tested in combination with azidothymidine (AZT) and acyclovir (ACY) in vitro. The rationale for such combinations was based on the antiviral and immunomodulatory properties exhibited by ACE-M. In addition, the observed antiviral effects of ACE-M against human immunodeficiency virus type 1 (HIV-1) and other enveloped viruses appear to be related to modification of the glycosylation of viral glycoproteins. Therefore, the inhibitory effect of ACE-M does not overlap with that of AZT or ACY. The studies presented herein show that ACE-M combined with suboptimal noncytotoxic concentrations of AZT or ACY act synergistically to inhibit the replication of HIV-1 and herpes simplex virus type 1 (HSV-1), respectively. The median effect method was not applicable for analysis because the test compounds show mutually nonexclusive drug effects. For a meaningful evaluation and interpretation of the effects of drug combinations, the biological significance of combinations must be considered, that is, the protective effect of the combination, the noncytotoxicity of the combination, the mechanism(s) of action of the individual compounds comprising the combination, and so forth. With respect to effects on U1 cells latently infected with HIV-1, treatment with combinations of AZT and ACE-M does not potentiate virus replication.


Beta3-containing gamma-aminobutyric acidA receptors are not major targets for the amnesic and immobilizing actions of isoflurane.
Mice bearing an N265M point mutation in the gamma-aminobutyric acid (GABA)(A) receptor beta3 subunit resist various anesthetic effects of propofol and etomidate. They also require a 16% larger concentration of enflurane and a 21% larger concentration of halothane to abolish the withdrawal reflex than do wild-type mice. Using a Pavlovian test, we measured whether this mutation increased the concentration of isoflurane required to impair learning and memory relative to wild-type mice. We found that the concentration was not significantly increased. We also measured MAC (the minimum alveolar concentration required to eliminate movement in response to noxious stimulation in 50% of subjects). Isoflurane MAC for mutant mice (1.93% +/- 0.0.03%; mean +/- se; n = 14) was 17.0% larger than MAC for wild-type mice (1.65 +/- 0.04; n = 14; P < 0.001). Similarly, the cyclopropane MAC for mutant mice (27.6% +/- 0.55%; n = 16) was 13.6% larger than MAC for wild-type mice (24.3 +/- 0.46; n = 8; P < 0.01). The increase in MAC for cyclopropane was unexpected, because published reports find only minimal actions at alpha1beta2gamma2 GABA(A) receptors whereas isoflurane provides a large enhancement. Consistent with previous work on alpha1beta2gamma2 GABA(A) receptors, we found in Xenopus oocytes that 5 MAC cyclopropane enhanced the effect of GABA on alpha1beta2gamma2 GABA(A) receptors by only 76%, and by a nearly identical enhancement in alpha1beta3gamma2, and alpha6beta3gamma2 receptors. In contrast, a much smaller concentration of isoflurane (1 MAC) produced a 160% to 310% enhancement in these receptors. If, relative to isoflurane, cyclopropane minimally increases GABA-induced chloride currents at any GABA(A) receptor subtype, the present data for MAC are consistent with the notion that GABA(A) receptors do not mediate the immobility produced by inhaled anesthetics. The results of the present study indicate that beta3-containing gamma-aminobutyric acidA receptors do not mediate the amnesia produced by isoflurane and do not mediate, or only partially mediate, the immobility produced by inhaled anesthetics.


The Streptococcos suis sortases SrtB and SrtF are essential for disease in pigs.
The porcine pathogen Streptococcus suis colonizes the upper respiratory tracts of pigs, potentially causing septicaemia, meningitis and death, thus placing a severe burden on the agricultural industry worldwide. It is also a zoonotic pathogen that is known to cause systemic infections and meningitis in humans. Understanding how S. suis colonizes and interacts with its hosts is relevant for future strategies of drug and vaccine development. As with other Gram-positive bacteria, S. suis utilizes enzymes known as sortases to attach specific proteins bearing cell wall sorting signals to its surface, where they can play a role in host-pathogen interactions. The surface proteins of bacteria are often important in adhesion to and invasion of host cells. In this study, markerless in-frame deletion mutants of the housekeeping sortase srtA and the two pilus-associated sortases, srtB and srtF, were generated and their importance in S. suis infections was investigated. We found that all three of these sortases are essential to disease in pigs, concluding that their cognate-sorted proteins may also be useful in protecting pigs against infection.


Bilateral thalamic stroke due to occlusion of the artery of Percheron in a patient with a patent foramen ovale.
The artery of Percheron (AOP) is a rare vascular variant in which a single dominant thalamoperforating artery arises from the P1 segment and bifurcates to supply both paramedian thalami. Occlusion of this uncommon vessel results in a characteristic pattern of bilateral paramedian thalamic infarcts with or without mesencephalic infarctions. We report a case of a 37-year-old man with acute bilateral thalamic infarcts. The scans revealed symmetric bilateral hyperintense paramedian thalamic lesions consistent with an acute ischemic event. The posterior circulation was patent including the tip of the basilar artery and both posterior cerebral arteries, making the case compatible with occlusion of the AOP. This type of infarct is associated with embolic phenomena, and further evaluation revealed a patent foramen ovale as the source of emboli in the cerebrovascular circulation. The occlusion of the AOP is a rare cause of coma in young patients, and early recognition of this rare disease entity may lead to more favorable outcomes.


Hospitalization in patients with systemic lupus erythematosus at Tawam Hospital, United Arab Emirates (UAE): Rates, causes, and factors associated with length of stay.
Systemic lupus erythematosus (SLE) is a relapsing and remitting multiorgan disease associated with significant morbidity and mortality. The survival rate of patients with SLE has recently improved, which was associated with increased morbidity and hospitalization rates. Therefore, this study aimed to examine the rate and causes of hospitalization in patients with SLE and explore factors associated with increased length of stay (LOS). Patients who visited rheumatology clinics (Tawam hospital, United Arab Emirates (UAE)) and fulfilled the American College of Rheumatology (ACR) SLE criteria were identified. Retrospective charts were reviewed to determine previous admissions. Demographic data, reason for hospitalization, duration of hospitalization, intensive care unit (ICU) admission, number of specialist consultations, medications used, and SLE characteristics at time of admission were collected. The hospitalization rate was calculated as the number of hospitalized patients divided by the total number of patients with the disease. We performed multivariable regression analysis for factors associated with increased LOS. A total of 91 patients with SLE (88 women and 3 men) met the inclusion criteria with a mean disease duration of 10.2 years (SD 5.5). A total of 222 admissions were identified, and 66 of 91 patients were admitted at least once. The mean crude hospitalization rate calculated was 29.8%. The primary reason for admission was pregnancy (29%), SLE activity (24%), and infection (20%). When combining primary and secondary reasons, the proportion of admissions due to SLE activity increased to 32%. The mean LOS was 5.9 (SD 6.0) days. About 7% of admitted patients required ICU admission. In multivariable analysis, patients with lupus nephritis, complications during hospitalization, and increased number of specialists consultations and who were admitted to ICU and started new medication were all associated with increased LOS. A significant proportion of patients with SLE were hospitalized during their disease course. The hospitalization rate in this study appears to be higher than those reported elsewhere. Disease flare is the leading cause of admission in patients with SLE in this relatively young cohort. Lupus nephritis has been found to be significantly related to longer LOS. Measurements taken to reduce the incidence and severity of flares would likely decrease hospitalization rate and LOS in patients with SLE.


Ketogenic diet combined with antioxidant N-acetylcysteine inhibits tumor growth in a mouse model of anaplastic thyroid cancer.
Anaplastic thyroid cancer is an aggressive and fatal malignancy. Many advanced cancers are characterized by glucose dependency, leading to oxidative stress and cellular proliferation. Therefore, we sought to determine if a low glucose environment (in vitro) or a ketogenic diet (in vivo) could inhibit anaplastic thyroid cancer tumor growth when combined with the antioxidant N-acetylcysteine. In vivo, nude mice were injected with the anaplastic thyroid cancer cell line 8505C (n = 6/group). Group 1 was fed a standard diet; Group 2 was fed a ketogenic diet; Group 3 was given standard diet with N-acetylcysteine (40 mM in the drinking water); and Group 4 was fed ketogenic diet with N-acetylcysteine. Tumor volumes, ketones, and glucose were measured. H&E stains and immunohistochemistry for Ki-67 and Caspase 3 were performed on the tumors. In vitro, 8505C cells were cultured in high glucose (25 mM), low glucose (3 mM), high glucose plus N-acetylcysteine (200 uM), or low glucose plus N-acetylcysteine for 96 hours. We performed CyQUANT proliferation (Thermo Fisher Scientific, Waltham, MA), Seahorse glycolytic stress (Agilent, Santa Clara, CA), and reactive oxidative stress assays. Ketogenic diet plus N-acetylcysteine decreased in vivo tumor volume compared to standard diet (22.5 ± 12.4 mm3 vs 147 ± 54.4 mm3, P < .05) and standard diet plus N-acetylcysteine (P < .05). Blood ketone levels were significantly higher for the mice in the ketogenic diet group compared to standard diet (1.74 mmol/L vs 0.38 mmol/L at week 5, P < .001). However, blood glucose levels were not significantly different between ketogenic diet and standard diet groups. Cells cultured in low glucose plus N-acetylcysteine had significantly reduced proliferation compared to high glucose (98.1 ± 5.0 relative fluorescence units vs 157.8 ± 2.1 relative fluorescence units, P < .001). Addition of N-acetylcysteine to low glucose lowered glycolysis function compared to high glucose (39.0 ± 2.2 mpH/min/cell vs 89.1 ± 13.2 mpH/min/cell, P < .001) and high glucose plus N-acetylcysteine (37.4 ± 2.5 mpH/min/cell vs 70.3 ± 3.3 mpH/min/cell, P < .001). Low glucose plus N-acetylcysteine decreased reactive oxidative stress compared to high glucose (119 ± 34.7 relative fluorescence units vs 277 ± 16.0 relative fluorescence units, P = .014). The combination of a ketogenic diet or glucose restriction with the antioxidant- N-acetylcysteine significantly reduced tumor growth in vivo and in vitro. Further studies are warranted to explore these metabolic therapies in anaplastic thyroid cancer.


Angioplasty does not affect subsequent operative renal artery revascularization.
Although increased application of percutaneous renal artery angioplasty and stenting has facilitated nonoperative renal revascularization, patient outcomes after failed angioplasty are not established. Renal artery revascularization was performed in 31 patients (38 arteries) from 1993 to 1999. Twenty patients underwent primary surgical repair, and 11 patients underwent secondary reconstruction after angioplasty (n = 7) or angioplasty and stenting (n = 4). Before operation, all patients had severe hypertension (blood pressure 166+/-5.2/92 +/- 2.7 mm Hg) that required an average of 3.0 +/- 0.2 medications for control. In addition, 12 patients (primary 45% vs secondary 27%; P = NS) had evidence of renal insufficiency (creatinine > or =1.7 mg/dL). There was no difference between primary and secondary procedures in the length of hospital stay (12+/- 1.4 vs. 12+/-3.2 days; P = NS), major morbidity (10% vs. 18%; P = NS) or perioperative mortality (overall mortality 2 of 31; primary 5% vs secondary 9%; P = NS). The majority of patients demonstrated improvement or cure of hypertension (primary 94% vs secondary 90%; P = NS) and stable or decreased creatinine (primary 74% vs secondary 82%; P = not significant). Overall survival (mean follow-up 22+/-3.5 months) was 89%+/-5.7%. Although this surgical series does not address the true outcomes of renal artery angioplasty, the results suggest that renal artery angioplasty does not prejudice subsequent surgical outcomes in patients who are carefully followed after angioplasty.


Mitogen activated protein kinase activation and oxidant signaling in astrocytoma cells.
Presence of increased reactive oxygen species (ROS) has been observed in most high risk factors for brain tumor development. Our past study demonstrated that ROS could induce increased brain tumor cell proliferation. Growth effects of ROS may involve modifications of cellular proteins such as mitogen-activated protein kinases (MAPKs), which regulate cell proliferation. Here, we report effects of a ROS (hydrogen peroxide, H2O2) and an antioxidant (N-acetylcysteine, NAC) on MAPK activation in astrocytoma (U373-MG) cells. MAPKs are activated by phosphorylation that can be detected by Western blot analysis. The unphosphorylated/inactivated form of MAPK exhibits slower mobility on SDS-PAGE compared to the phosphorylated/activated form. Densitometric analysis was used to measure MAPK activation. Results indicate that H2O2 caused a dose and time-dependent increase in MAPK activation in astrocytoma cells. Furthermore, ROS-induced activation was almost completely suppressed by NAC. NAC also inhibited serum-induced MAPK activation indicating there may be an oxidant-sensitive component to serum-induced growth signaling. Modifications of MAPKs by H2O2 demonstrate that ROS-induced proliferation is via biochemical pathways similar to other known growth stimuli. Understanding of processes that link a proliferation signal (ROS) to cell proliferation can aid in the selection of therapy used to suppress brain tumor growth.


Characterization of herpes simplex virus 2 temperature-sensitive mutants whose lesions map in or near the coding sequences for the major DNA-binding protein.
By marker rescue with cloned herpes simplex virus 2 DNA fragments, we have mapped the temperature-sensitive mutations of a series of herpes simplex virus 2 mutants to a region of the herpes simplex virus 2 genome that lies within or near the coding sequences for the major DNA-binding protein, ICP8. In cells infected with certain of these mutants at the nonpermissive temperature, the association of the major DNA-binding protein with the cell nucleus was defective. In these cells, the DNA-binding protein accumulated in the cytoplasmic and the crude nuclear detergent wash fractions. At the permissive temperature, the maturation of the mutant ICP8 was similar to that of the wild-type viral protein. With the remainder of the mutants, the nuclear maturation of ICP8 was similar to that encoded by the wild-type virus at the nonpermissive and permissive temperatures as assayed by cell fractionation.


Ion-pair amphiphile: a neoteric substitute that modulates the physicochemical properties of biomimetic membranes.
Ion-pair amphiphiles (IPAs) are neoteric pseudo-double-tailed compounds with potential as a novel substitute of phospholipid. IPA, synthesized by stoichiometric/equimolar mixing of aqueous solution of hexadecyltrimethylammonium bromide (HTMAB) and sodium dodecyl sulfate (SDS), was used as a potential substituent of naturally occurring phospholipid, soylecithin (SLC). Vesicles were prepared using SLC and IPA in different ratios along with cholesterol. The impact of IPA on SLC was examined by way of surface pressure (π)-area (A) measurements. Associated thermodynamic parameters were evaluated; interfacial miscibility between the components was found to depend on SLC/IPA ratio. Solution behavior of the bilayers, in the form of vesicles, was investigated by monitoring the hydrodynamic diameter, zeta potential, and polydispersity index over a period of 100 days. Size and morphology of the vesicles were also investigated by electron microscopic studies. Systems comprising 20 and 40 mol % IPA exhibited anomalous behavior. Thermal behavior of the vesicles, as scrutinized by differential scanning calorimetry, was correlated with the hydrocarbon chain as well as the headgroup packing. Entrapment efficiency (EE) of the vesicles toward the cationic dye methylene blue (MB) was also evaluated. Vesicles were smart enough to entrap the dye, and the efficiency was found to vary with IPA concentration. EE was found to be well above 80% for some stable dispersions. Such formulations thus could be considered to have potential as novel drug delivery systems.


Efficient Delay Correction for Total-Body PET Kinetic Modeling Using Pulse Timing Methods.
Quantitative kinetic modeling requires an input function. A noninvasive image-derived input function (IDIF) can be obtained from dynamic PET images. However, a robust IDIF location (e.g., aorta) may be far from a tissue of interest, particularly in total-body PET, introducing a time delay between the IDIF and the tissue. The standard practice of joint estimation (JE) of delay, along with model fitting, is computationally expensive. To improve the efficiency of delay correction for total-body PET parametric imaging, this study investigated the use of pulse timing methods to estimate and correct for delay. Methods: Simulation studies were performed with a range of delay values, frame lengths, and noise levels to test the tolerance of 2 pulse timing methods-leading edge (LE) and constant fraction discrimination and their thresholds. The methods were then applied to data from 21 subjects (14 healthy volunteers, 7 cancer patients) who underwent a 60-min dynamic total-body 18F-FDG PET acquisition. Region-of-interest kinetic analysis was performed and parametric images were generated to compare LE and JE methods of delay correction, as well as no delay correction. Results: Simulations demonstrated that a 10% LE threshold resulted in biases and SDs at tolerable levels for all noise levels tested, with 2-s frames. Pooled region-of-interest-based results (n = 154) showed strong agreement between LE (10% threshold) and JE methods in estimating delay (Pearson r = 0.96, P < 0.001) and the kinetic parameters vb (r = 0.96, P < 0.001), Ki (r = 1.00, P < 0.001), and K1 (r = 0.97, P < 0.001). When tissues with minimal delay were excluded from pooled analyses, there were reductions in vb (69.4%) and K1 (4.8%) when delay correction was not performed. Similar results were obtained for parametric images; additionally, lesion Ki contrast was improved overall with LE and JE delay correction compared with no delay correction and Patlak analysis. Conclusion: This study demonstrated the importance of delay correction in total-body PET. LE delay correction can be an efficient surrogate for JE, requiring a fraction of the computational time and allowing for rapid delay correction across more than 106 voxels in total-body PET datasets.


Role of a holo-insertase complex in the biogenesis of biophysically diverse ER membrane proteins.
Mammalian membrane proteins perform essential physiologic functions that rely on their accurate insertion and folding at the endoplasmic reticulum (ER). Using forward and arrayed genetic screens, we systematically studied the biogenesis of a panel of membrane proteins, including several G-protein coupled receptors (GPCRs). We observed a central role for the insertase, the ER membrane protein complex (EMC), and developed a dual-guide approach to identify genetic modifiers of the EMC. We found that the back of sec61 (BOS) complex, a component of the 'multipass translocon', was a physical and genetic interactor of the EMC. Functional and structural analysis of the EMC•BOS holocomplex showed that characteristics of a GPCR's soluble domain determine its biogenesis pathway. In contrast to prevailing models, no single insertase handles all substrates. We instead propose a unifying model for coordination between the EMC, multipass translocon, and Sec61 for biogenesis of diverse membrane proteins in human cells.


Immunotoxins up to the present day.
Immunotoxins consist of monoclonal or polyclonal antibodies conjugated to bacterial or plant toxins. The toxins used are typically of the A-B type in which a toxic A chain is coupled to a B chain responsible for cell binding and facilitation of A chain entry into the cytosol. Two broad strategies have been followed: coupling intact toxins, or A chains alone, to antibodies. This review examines current progress in in vitro and in vivo research, including recent clinical studies, concentrating principally on ricin or ricin A chain conjugates. The future role of conjugates using membrane-acting toxins, immunolysins, is also discussed.


Opioid sparing effect of low dose ketamine in patients with intravenous patient-controlled analgesia using fentanyl after lumbar spinal fusion surgery.
The opioid sparing effect of low dose ketamine is influenced by bolus dose, infusion rate, duration of infusion, and differences in the intensity of postoperative pain. In this study, we investigated the opioid sparing effect of low dose ketamine in patients with intravenous patient-controlled analgesia (PCA) using fentanyl after lumbar spinal fusion surgery, which can cause severe postoperative pain. Sixty patients scheduled for elective lumbar spinal fusion surgery were randomly assigned to receive one of three study medications (K1 group: ketamine infusion of 1 µg/kg/min following bolus 0.5 mg/kg, K2 group: ketamine infusion of 2 µg/kg/min following bolus 0.5 mg/kg, saline infusion following bolus of saline). Continuous infusion of ketamine began before skin incision intraoperatively, and continued until 48 h postoperatively. For postoperative pain control, patients were administered fentanyl using IV-PCA (bolus dose 15 µg of fentanyl, lockout interval of 5 min, no basal infusion). For 48 h postoperatively, the total amount of fentanyl consumption, postoperative pain score, adverse effects and patients' satisfaction were evaluated. The total amount of fentanyl consumption was significantly lower in the K2 group (474 µg) compared to the control group (826 µg) and the K1 group (756 µg) during the 48 h after surgery. Pain scores at rest or with movement, the incidence of adverse events and patient satisfaction were not significantly different among the groups. Low-dose ketamine at 2 µg/kg/min following bolus 0.5 mg/kg significantly reduced the total amount of fentanyl consumption during the 48 h after lumbar spinal fusion surgery without increasing adverse effects.


Generation and Evolution of High-Mach-Number Laser-Driven Magnetized Collisionless Shocks in the Laboratory.
We present the first laboratory generation of high-Mach-number magnetized collisionless shocks created through the interaction of an expanding laser-driven plasma with a magnetized ambient plasma. Time-resolved, two-dimensional imaging of plasma density and magnetic fields shows the formation and evolution of a supercritical shock propagating at magnetosonic Mach number M_{ms}≈12. Particle-in-cell simulations constrained by experimental data further detail the shock formation and separate dynamics of the multi-ion-species ambient plasma. The results show that the shocks form on time scales as fast as one gyroperiod, aided by the efficient coupling of energy, and the generation of a magnetic barrier between the piston and ambient ions. The development of this experimental platform complements present remote sensing and spacecraft observations, and opens the way for controlled laboratory investigations of high-Mach number collisionless shocks, including the mechanisms and efficiency of particle acceleration.


Acute myeloid leukemia and myelodysplastic syndromes following essential thrombocythemia treated with hydroxyurea: high proportion of cases with 17p deletion.
Treatment with alkylating agents or radiophosphorous (32P) has been shown to carry a certain leukemogenic risk in myeloproliferative disorders (MPDs), including essential thrombocytemia (ET). The leukemogenic risk associated to treatment with hydroxyurea in ET, on the other hand, is generally considered to be relatively low. Between 1970 and 1991, we diagnosed ET in 357 patients, who were monitored until 1996. One or several therapeutic agents had been administered to 326 patients, including hydroxyurea (HU) in 251 (as only treatment in 201), pipobroman in 43, busulfan in 41, and 32P in 40. With a median follow-up duration of 98 months, 17 patients (4.5%) had progressed to acute myeloid leukemia (AML; six cases) or myelodysplastic syndrome (MDS; 11 cases). Fourteen of these patients had received HU, as sole treatment in seven cases, and preceded or followed by other treatment in seven cases, mainly pipobroman (five cases). The remaining three leukemic progressions occurred in patients treated with 32P (two cases) and busulfan (one case). The incidence of AML and MDS after treatment, using 32P alone and 32P with other agents, busulfan alone and with other agents, HU alone and with others agents, and pipobroman alone and with other agents was 7% and 9%, 3% and 17%, 3.5% and 14%, and 0% and 16%, respectively. Thirteen of 17 patients who progressed to AML or MDS had successful cytogenetic analysis. Seven of them had rearrangements of chromosome 17 (unbalanced translocation, partial or complete deletion, isochromosome 17q) that resulted in 17p deletion. They also had a typical form of dysgranulopoiesis combining pseudo Pelger Hüet hypolobulation and vacuoles in neutrophils, and p53 mutation, as previously described in AML and MDS with 17p deletion. Those seven patients had all received HU, as the only therapeutic agent in three, and followed by pipobroman in three. The three patients who had received no HU and progressed to AML or MDS had no 17p deletion. A review of the literature found cytogenetic analysis in 35 cases of AML and MDS occurring after ET, 11 of whom had been treated with HU alone. Five of 35 patients had rearrangements that resulted in 17p deletion. Four of them had been treated with HU alone. These results show that treatment with HU alone is associated with a leukemic risk of approximately 3.5%. A high proportion of AML and MDS occurring in ET treated with HU (alone or possibly followed by pipobroman) have morphologic, cytogenetic, and molecular characteristics of the 17p- syndrome. These findings suggest that widespread and prolonged use of HU in ET may have to be reconsidered in some situations, such as asymptomatic ET.


Antimetabolites produced by microorganisms. XII (S)-alanyl-3-[alpha-(S)-chloro-3-(S)-hydroxy 2-oxo-3-azetidinylmethyl]-(S)-alanine, a new beta-lactam containing natural product.
(S)-Alanyl-3-[alpha-(S)-chloro-3-(S)-hydroxy-2-oxo-3-azetidinylmethyl]-(S)-alanine was isolated from a fermentation broth of an unidentified Streptomyces species 372 A. The structure was determined by single crystal X-ray diffraction analysis. The substance inhibits the growth of several strains of gram-positive and gram-negative bacteria in a chemically defined medium but growth inhibition is relieved by addition of L-glutamine to the medium.


When side matters: hemispheric processing and the visual specificity of emotional memories.
Previous studies have shown that the right hemisphere processes the visual details of objects and the emotionality of information. These two roles of the right hemisphere have not been examined concurrently. In the present study, the authors examined whether right hemisphere processing would lead to particularly good memory for the visual details of emotional stimuli. Participants viewed positive, negative, and neutral objects, displayed to the left or right of a fixation cross. Later, participants performed a recognition task in which they evaluated whether items were "same" (same visual details), "similar" (same verbal label, different visual details), or "new" (unrelated) in comparison with the studied objects. Participants remembered the visual details of negative items well, and this advantage in memory specificity was particularly pronounced when the items had been presented directly to the right hemisphere (i.e., to the left of the fixation cross). These results suggest that there is an episodic memory benefit conveyed when negative items are presented directly to the right hemisphere, likely because of the specialization of the right hemisphere for processing both visual detail and negatively valenced emotional information.


Small interfering RNA in drug metabolism and transport.
RNA interference (RNAi) is a powerful technique that utilizes RNA molecules to specifically knock down the expression of targeted gene at posttranscriptional level. These small interfering RNAs (siRNAs) not only have broad application to basic biomedical research but may be developed as therapeutic agents. Drug-metabolizing enzymes (DMEs) and drug transporters (DTs) are molecular determinants of pharmacokinetic property of a drug. Transcriptional gene expression of DMEs and DTs is controlled by xenobiotic-sensing nuclear receptors (NRs). Because of complexity in studying the function of individual DMEs, DTs and NRs, siRNAs can be an excellent addition to chemical inhibitors and inhibitory antibodies in delineating their specific roles in drug metabolism and transport, gene regulation, and drug-drug interactions. RNAi may be employed to modulate DT expression to overcome multidrug resistance. Recent studies using RNAi to silence gene expression of specific DME, DT and NR, and the impact on drug metabolism and transport are discussed in this review. Concerns remain about the efficiency, specificity, and off-target effects when interpreting data obtained from RNAi studies. Furthermore, potential role for endogenous siRNAs, microRNA (miRNA) molecules, in controlling the posttranscriptional gene regulation of DMEs, DTs and NRs is discussed.


Extending half-life by indirect targeting of the neonatal Fc receptor (FcRn) using a minimal albumin binding domain.
The therapeutic and diagnostic efficiency of engineered small proteins, peptides, and chemical drug candidates is hampered by short in vivo serum half-life. Thus, strategies to tailor their biodistribution and serum persistence are highly needed. An attractive approach is to take advantage of the exceptionally long circulation half-life of serum albumin or IgG, which is attributed to a pH-dependent interaction with the neonatal Fc receptor (FcRn) rescuing these proteins from intracellular degradation. Here, we present molecular evidence that a minimal albumin binding domain (ABD) derived from streptococcal protein G can be used for efficient half-life extension by indirect targeting of FcRn. We show that ABD, and ABD recombinantly fused to an Affibody molecule, in complex with albumin does not interfere with the strictly pH-dependent FcRn-albumin binding kinetics. The same result was obtained in the presence of IgG. An in vivo study performed in rat confirmed that the clinically relevant human epidermal growth factor 2 (HER2)-targeting Affibody molecule fused to ABD has a similar half-life and biodistribution profile as serum albumin. The proof-of-concept described may be broadly applicable to extend the in vivo half-life of short lived biological or chemical drugs ultimately resulting in enhanced therapeutic or diagnostic efficiency, a more favorable dosing regimen, and improved patient compliance.


Patient-reported Outcomes, Functional Assessment, and Utility Values in Glaucoma.
In clinical glaucoma research, the measurement of patient reported outcomes, functional assessment of disability, and health economic impact is critical. However, valid, time-efficient and comprehensive tools are not available and several current instruments lack in the appropriate precision for measuring the various dimensions of glaucoma-related quality of life (QoL), including functioning and mobility. Furthermore, statistical methods are inconsistently and sometimes incorrectly used in otherwise sound clinical studies. Standardizing and improving methods of patient-centered data collection and analysis in glaucoma studies are imperative. This paper outlines recommendations and provides a discussion of some of the pertinent issues relating to the optimization of patient-reported outcomes research in glaucoma.


Management of Scaphotrapeziotrapezoid Osteoarthritis: A Critical Analysis Review.
» The scaphotrapeziotrapezoid (STT) joint is a common location of degenerative osteoarthritis (OA) in the wrist.» STT OA may occur in isolation or with concomitant thumb carpometacarpal joint (CMCJ) OA or scapholunate advanced collapse (SLAC) and other wrist OA patterns.» Nonoperative management strategies include activity modification, bracing, anti-inflammatory medications, occupational therapy, and injections.» Surgical treatment may be determined by whether the STT OA occurs in isolation or with concomitant thumb CMCJ or SLAC OA and includes STT arthrodesis, distal scaphoid excision with or without interposition or implant arthroplasty, trapeziectomy with proximal trapezoid excision, four-corner fusion in combination with scaphoid/radial column excision, and proximal row carpectomy.


Etiology and Treatment of Intraoperative Hyperkalemia During Posterior Spinal Fusion in an Adolescent.
Hyperkalemia, defined as a serum or plasma potassium greater than 5.5 mEq/L, while an uncommon occurrence in children, is a serious medical problem that warrants immediate attention as it can result in serious cardiac arrhythmias and death. Although hyperkalemia may occur in the critically ill patient or in the setting of renal failure and insufficiency, there are limited reports of its occurrence during intraoperative care. The authors report a previously healthy, 18-year-old patient, who developed hyperkalemia intraoperatively during posterior spinal fusion to treat adolescent idiopathic scoliosis. The potential etiologies of hyperkalemia are reviewed, a differential diagnosis for the possible etiologies presented, and treatment modalities discussed.


Diagnostic and prognostic value of Ischemia-modified albumin in patients with Crimean-Congo hemorrhagic fever.
The aim of this study was to determine the diagnostic and prognostic significance of ischemia modified albumin (IMA) levels in patients with Crimean-Congo hemorrhagic fever (CCHF). This retrospective study was conducted with patients with CCHF. IMA levels in patients with CCHF were determined using the rapid colorimetric method. IMA levels of CCHF patients were significantly higher compared with the control group (P = 0.0001). At an IMA cut-off point of 0.555 ABSU (absorbance units), sensitivity was 65.1%, specificity 82.5%, positive predictive values (PPV) 82.5%, and negative predictive values (NPV) 65.1%. IMA levels of patients with hemorrhage were significantly higher compared with patients without hemorrhage (P = 0.005). IMA has been validated as both a new and sensitive ischemia and oxidative stress biomarker. In addition to its diagnostic significance, IMA investigated in CCHF patients at time of arrival may be an important marker with its prognostic role in determining in the early stage whether the disease will follow a hemorrhagic course.


Doppler ultrasonographical findings in the differential diagnosis of post-partum thyroiditis.
To investigate the effects of haemodynamic indices on colour Doppler ultrasound in differential diagnosis in patients with postpartum thyroiditis and with Graves' disease. The cross-sectional study was conducted at the Endocrinology Polyclinic of Medical Park Hospital, Ordu, Turkey, from March 2017 to May 2018 and comprised patients referred from the Gynaecology Department for routine check-up after parturition within the first 6 months. The patients were divided into two groups. Group 1 had postpartum thyroiditis patients and Group 2 had those of Graves' disease. In both groups, parameters measured were peak systolic velocity, end-diastolic velocity and resistive index of the inferior thyroid artery with proper angle (45-60⁰) on colour Doppler ultrasound. Data was analysed using SPSS 20. Of the 42 subjects,18(42.85%) were in Group 1 and 24(57.14%) were in Group 2. Peak systolic velocity and end-diastolic velocity values of the inferior thyroid artery were higher in Group 2 compared to Group 1 (p<005). (p<0.05), while the resistive index value was significantly higher in Group 1 compared to Group 2. Due to its wide availability, the use of colour Doppler ultrasound parameters indicating parenchymal perfusion were found to be broadly useful in distinguishing between postpartum thyroiditis and Graves' disease.


A closed loop wireless power transmission system using a commercial RFID transceiver for biomedical applications.
This paper presents a standalone closed loop wireless power transmission system that is built around a commercial off-the-shelf (COTS) radio frequency identification (RFID) transceiver (MLX90121) operating at 13.56 MHz. It can be used for inductively powering implantable biomedical devices in a closed loop fashion. Any changes in the distance and misalignment between transmitter and receiver coils in near-field wireless power transmission can cause a significant change in the received power, which can cause either malfunction or excessive heat dissipation. RFID transceivers are often used open loop. However, their back telemetry capability can be utilized to stabilize the received voltage on the implant. Our measurements showed that the delivered power to the transponder was maintained at 1.48 mW over a range of 6 to 12 cm, while the transmitter power consumption changed from 0.3 W to 1.21 W. The closed loop system can also oppose voltage variations as a result of sudden changes in load current.


Excess prenatal corticosterone exposure results in albuminuria, sex-specific hypotension, and altered heart rate responses to restraint stress in aged adult mice.
Exposure to excess glucocorticoids programs susceptibility to cardiovascular and renal dysfunction in later life although the mechanisms have not been clearly elucidated. We administered corticosterone (CORT; 33 μg·kg(-1)·h(-1)) to pregnant mice for 60 h from embryonic day (E) 12.5. Prenatal CORT resulted in postnatal growth restriction and reduced nephron endowment at postnatal day 30 in both male and female offspring. The reduction in nephron number was associated with increased expression of apoptotic markers in the kidney at E14.5. In offspring of both sexes at 12 mo of age, there were no differences in kidney weights, urine output, or urinary sodium excretion; however, prenatal CORT exposure increased the urinary albumin/creatinine ratio and 24-h urinary albumin excretion. Surprisingly, at 12 mo male but not female offspring exposed to prenatal CORT were hypotensive, with mean arterial blood pressures ∼10 mmHg lower than untreated controls (P < 0.001). Finally, we examined how offspring responded to a renal or cardiovascular challenge (saline load or restraint stress). When given 0.9% NaCl as drinking water for 7 days, there were no differences in blood pressures or urinary parameters between groups. Restraint stress (15 min) caused a tachycardic response in all animals; however the increase in heart rate was not sustained in male offspring exposed to CORT (P < 0.01), suggesting that autonomic control of cardiovascular function may be altered. These data demonstrate that excess prenatal CORT impairs kidney development and increases the risk of cardiovascular dysfunction especially in males.


Proteomic screen in the simple metazoan Hydra identifies 14-3-3 binding proteins implicated in cellular metabolism, cytoskeletal organisation and Ca2+ signalling.
14-3-3 proteins have been implicated in many signalling mechanisms due to their interaction with Ser/Thr phosphorylated target proteins. They are evolutionarily well conserved in eukaryotic organisms from single celled protozoans and unicellular algae to plants and humans. A diverse array of target proteins has been found in higher plants and in human cell lines including proteins involved in cellular metabolism, apoptosis, cytoskeletal organisation, secretion and Ca2+ signalling. We found that the simple metazoan Hydra has four 14-3-3 isoforms. In order to investigate whether the diversity of 14-3-3 target proteins is also conserved over the whole animal kingdom we isolated 14-3-3 binding proteins from Hydra vulgaris using a 14-3-3-affinity column. We identified 23 proteins that covered most of the above-mentioned groups. We also isolated several novel 14-3-3 binding proteins and the Hydra specific secreted fascin-domain-containing protein PPOD. In addition, we demonstrated that one of the 14-3-3 isoforms, 14-3-3 HyA, interacts with one Hydra-Bcl-2 like protein in vitro. Our results indicate that 14-3-3 proteins have been ubiquitous signalling components since the start of metazoan evolution. We also discuss the possibility that they are involved in the regulation of cell numbers in response to food supply in Hydra.


The ubiquitin C-terminal hydrolase UCH-L1 promotes bacterial invasion by altering the dynamics of the actin cytoskeleton.
Invasion of eukaryotic target cells by pathogenic bacteria requires extensive remodelling of the membrane and actin cytoskeleton. Here we show that the remodelling process is regulated by the ubiquitin C-terminal hydrolase UCH-L1 that promotes the invasion of epithelial cells by Listeria monocytogenes and Salmonella enterica. Knockdown of UCH-L1 reduced the uptake of both bacteria, while expression of the catalytically active enzyme promoted efficient internalization in the UCH-L1-negative HeLa cell line. The entry of L. monocytogenes involves binding to the receptor tyrosine kinase Met, which leads to receptor phosphorylation and ubiquitination. UCH-L1 controls the early membrane-associated events of this triggering cascade since knockdown was associated with altered phosphorylation of the c-cbl docking site on Tyr1003, reduced ubiquitination of the receptor and altered activation of downstream ERK1/2- and AKT-dependent signalling in response to the natural ligand Hepatocyte Growth Factor (HGF). The regulation of cytoskeleton dynamics was further confirmed by the induction of actin stress fibres in HeLa expressing the active enzyme but not the catalytic mutant UCH-L1(C90S) . These findings highlight a previously unrecognized involvement of the ubiquitin cycle in bacterial entry. UCH-L1 is highly expressed in malignant cells that may therefore be particularly susceptible to invasion by bacteria-based drug delivery systems.


Diagnostic Utility of PD-L1 Expression in Lung Adenocarcinoma: Immunohistochemistry and RNA In Situ Hybridization.
Programmed death receptor and programmed death ligand (PD-L1) are immunoregulatory proteins. Nonsmall cell lung cancer bypasses the immune system through the induction of protumorigenic immunosuppressive changes. The better understanding of immunology and antitumor immune responses has brought the promising development of novel immunotherapy agents like programmed death receptor checkpoint inhibitors. The aim of this study was to investigate the expression of PD-L1 in lung adenocarcinoma (ADC), comparing 2 different technologies: immunohistochemistry (IHC) by 2 methods versus RNA in situ hybridization (RISH). In total, 20 cases of ADC of the lung and 4 samples of metastatic colon ADC were selected. Evaluation of PD-L1 expression was performed by IHC and RISH. RISH was performed using RNAscope. Both methods were scored in tumor cells and quantified using combined intensity and proportion scores. Eight of 20 (40%) lung ADC and 2 of 4 (50%) colon ADC were positive for PD-L1 with Cell Signaling IHC, and 65% lung ADC were positive by Dako IHC (13/20). All 4 cases of colon ADC were negative. When evaluated by RISH, 12 lung ADC (60%) and 1 colon ADC (25%) were PD-L1 positive. RNAscope probes provide sensitive and specific detection of PD-L1 in lung ADC. Both IHC methods (Cell Signaling and Dako) show PD-L1 expression, with the Dako method more sensitive (40% vs. 65%). This study illustrates the utility of RISH and Cell Signaling IHC as complementary diagnostic tests, and Food and Drug Administration approved Dako IHC as a companion diagnostic test.


Assessment of dermal exposure to bitumen condensate among road paving and mastic crews with an observational method.
To assess dermal exposure to bitumen condensate among road pavers and indoor mastic workers in multiple crews using a semi-quantitative observational method [DeRmal Exposure Assessment Method (DREAM)]. Two skilled observers assessed dermal exposure to bitumen condensate among 85 asphalt workers from 12 crews from nine companies active within four European countries using the DREAM methodology, which produces an estimate of exposure expressed in dimensionless DREAM units. Both observers independently evaluated each crew member's job (N = 14 jobs) for road paving and mastic applications. Potential and actual dermal exposures were estimated for hands and for the rest of the body separately, taking into account the effect of protective clothing. To evaluate the reproducibility of the observational method intra-class correlation coefficients (ICCs) were estimated. The exposures in DREAM units were modelled using linear mixed models to estimate average relative scores for each job. Correlations between dermal exposure parameters were evaluated by estimating Pearson correlation coefficients. A total of 170 observations were completed by two observers independently (n = 118 and n = 52 for 59 road pavers and 26 mastic workers, respectively) in 11 days. The mean ICCs (for potential and actual exposure in DREAM units) varied between 0.74 and 0.80 with values for actual units being slightly higher. Geometric mean potential dermal exposure units of mastic workers were higher than for road pavers (factor 3 for hands and factor 4 for rest of the body). Differences for actual dermal exposure units were smaller for hands (factor 2) and larger for actual exposure units of rest of the body (factor 5). Differences in dermal exposure at the hands between jobs within a paving crew were much larger than between jobs within a mastic crew. Within paving crews, a consistent pattern for all exposure units emerged with 'screed man' and 'raker' as the two highest exposed jobs. Within mastic crews, 'driver dumper truck' and 'spreader of mastic' were scored as the two jobs with the highest exposure units. Potential and actual exposure units were highly correlated. Hands were more profoundly exposed than the rest of the body, with transfer from contaminated surface to the hands as the most important route. DREAM observations were reproducible and showed a consistent dermal exposure pattern among the observed crews. The study provided a clear picture of dermal exposure among road pavers and indoor mastic workers, with the mastic workers being considerably more highly exposed. The most important route of exposure appeared to be transferred from contaminated surfaces to the hands.


Magnetic and Optical Field Multi-Assisted Li-O2 Batteries with Ultrahigh Energy Efficiency and Cycle Stability.
The photoassisted lithium-oxygen (Li-O2 ) system has emerged as an important direction for future development by effectively reducing the large overpotential in Li-O2 batteries. However, the advancement is greatly hindered by the rapidly recombined photoexcited electrons and holes upon the discharging and charging processes. Herein, a breakthrough is made in overcoming these challenges by developing a new magnetic and optical field multi-assisted Li-O2 battery with 3D porous NiO nanosheets on the Ni foam (NiO/FNi) as a photoelectrode. Under illumination, the photogenerated electrons and holes of the NiO/FNi photoelectrode play a key role in reducing the overpotential during discharging and charging, respectively. By introducing the external magnetic field, the Lorentz force acts oppositely on the photogenerated electrons and holes, thereby suppressing the recombination of charge carriers. The magnetic and optical field multi-assisted Li-O2 battery achieves an ultralow charge potential of 2.73 V, a high energy efficiency of 96.7%, and good cycling stability. This external magnetic and optical field multi-assisted technology paves a new way of developing high-performance Li-O2 batteries and other energy storage systems.


Safety Assessment of Acer tegmentosum Maxim. Water Extract: General Toxicity Studies in Sprague-Dawley Rats and Beagle Dogs With Re-evaluation of Genotoxic Potentials.
Acer t egmentosum Maxim., commonly known as Manchurian stripe maple, is a deciduous tree belonging to the family of Aceraceae and has been traditionally used in folk medicine for its remedial effects in liver diseases and traumatic bleedings. With a growing body of experimental evidence for its pharmacological efficacies, such as neuroprotective, hepatoprotective, antioxidant, and anti-inflammatory activities, A. tegmentosum has gradually gained popularity as a health supplement and functional food. However, the large part of essential toxicity information still remained lacking despite the possibility of mutagenic potentials as previously suggested, posing safety concerns for human consumption. In this study, we evaluated 90-day repeated oral toxicity of A. tegmentosum Maxim. water extract (ATWE) in SD rats with acute toxicity assessment in beagle dogs, and reevaluated genotoxicity using a combination of in vitro and in vivo assays. During the oral study period, ATWE did not cause toxicity-related clinical signs and mortality in rodents without adverse effects observed in the analysis of hematology, serum biochemistry, and histopathology, establishing >5,000 mg/kg BW as the NOAEL. In addition, doses up to 5,000 mg/kg BW did not cause acute toxicity in beagle dogs. When assessed for genotoxicity using bacterial reverse mutation, chromosome aberration, and micronucleus formation, ATWE showed lack of mutagenicity and clastogenicity. These results demonstrated that AWTE was safe in the present preclinical study for systemic toxicity and genotoxicity at the tested doses, providing a guideline for safe use in humans.


Hippocampal expression of murine IL-4 results in exacerbation of amyloid deposition.
Pro-inflammatory stimuli, including cytokines like Interleukin-1β, Interleukin-6 and Interferon-γ, in the brain have been proposed to exacerbate existing Alzheimer's disease (AD) neuropathology by increasing amyloidogenic processing of APP and promoting further Aβ accumulation in AD. On the other hand, anti-inflammatory cytokines have been suggested to be neuroprotective by reducing neuroinflammation and clearing Aβ. To test this hypothesis, we used adeno-associated virus serotype 1 (AAV2/1) to express an anti-inflammatory cytokine, murine Interleukin-4 (mIL-4), in the hippocampus of APP transgenic TgCRND8 mice with pre-existing plaques. mIL-4 expression resulted in establishment of an "M2-like" phenotype in the brain and was accompanied by exacerbated Aβ deposition in TgCRND8 mice brains. No change in holo APP or APP C terminal fragment or phosphorylated tau levels were detected in mIL-4 expressing CRND8 cohorts. Biochemical analysis shows increases in both SDS soluble and insoluble Aβ. mIL-4 treatment attenuates soluble Aβ40 uptake by microglia but does not affect aggregated Aβ42 internalization by microglia or soluble Aβ40 internalization by astrocytes. Short term focal mIL-4 expression in the hippocampus leads to exacerbation of amyloid deposition in vivo, possibly mediated by acute suppression of glial clearance mechanisms. Given that recent preclinical data from independent groups indicate engagement of the innate immune system early on during disease pathogenesis may be beneficial, our present study strongly argues for a cautious re-examination of unwarranted side-effects of anti-inflammatory therapies for neurodegenerative diseases, including AD.


Vitamin E.
The term vitamin E is used to describe eight lipophilic, naturally occurring compounds that include four tocopherols and four tocotrienols designated as alpha-, beta-, gamma-, and delta-. The most well-known function of vitamin E is that of a chain-breaking antioxidant that prevents the cyclic propagation of lipid peroxidation. Despite its antioxidant function, dietary vitamin E requirements in humans are limited only to alpha-tocopherol because the other forms of vitamin E are poorly recognized by the hepatic alpha-tocopherol transfer protein (TTP), and they are not converted to alpha-tocopherol by humans. In attempts to gain a better understanding of vitamin E's health benefits, the molecular regulatory mechanisms of vitamin E have received increased attention. Examples of these mechanisms include: (1) the role of the hepatic alpha-TTP in preferentially secreting alpha-tocopherol into the plasma, (2) phase I and phase II metabolism of vitamin E and the potential impact for drug-vitamin E interactions, and (3) the regulation of biliary excretion of vitamin E by ATP-binding cassette protein(s). It is expected that the continued studies of these regulatory pathways will provide new insights into vitamin E function from which additional human health benefits will evolve.


The future of plasma derivatives. 17-18 April, 2000, Brussels, Belgium.
This two-day meeting was organised by Life Sciences Division of IBC Global Conferences Limited. It was attended by an international, multidisciplinary group of experts with differing expertise and perspectives on the provision of therapeutic products derived from human plasma. The delegates ranged from those involved with the collection of plasma donations, the scientists dedicated to improving the screening tests for virus markers, quality assurance personnel, clinicians, marketing and business managers, members of regulatory agencies in Europe and the Food and Drug Administration (FDA), providers of services to the industry and last, but not least, patient groups were represented. Fifteen to twenty countries were represented by the delegates and speakers. Although it might seem that the background and expertise of the audience was so wide that the lectures would have to be superficial, this was not the case. The whole process from plasma (or blood) collection to injection of a product into a patient was already well-known to senior members of the various organisations present and there is a tremendous amount of inter-dependence which enhances the necessary level of understanding. Nevertheless, the speakers were able to expand the knowledge of the delegates by their lucid and detailed presentations. The issues associated with development and implementation of new technologies, such as new NAT tests and different virus inactivation processes, were fully described. The spotlight also fell on the effects that changing patterns of demand for individual therapeutic proteins are having and will continue to have on the economics of the industry and on patient groups. Unlike other manufactured products, the raw material is the same for all plasma-derived products. Demand for all these products is limited by the availability of the donated plasma. Ideal demand for economic reasons is a balance of the extractable proteins in the donation. The implementation of European regulations was reviewed from the industry and from the regulators. Flexibility shown by the FDA in responding to the shortages of iv. immunoglobulins (Ig) was described, particularly with regard to clinical evaluation. Finally, the rationale for the need to revise the CPMP guidelines for clinical evaluation of certain plasma products was described as were the difficulties to be expected with implementation.


[Influence of different types of ionizing radiation on fatty acid profiles of cell lipids in microscopic fungi with radioadaptive properties].
The fatty acid profiles of strains of dark-pigmented (melanin-containing) species Cladosporium cladosporioides and Hormoconis resinae, and light-pigmented Aspergillus versicolor and Paecilomyces lilacinus, showing radioadaptive properties in comparison with control strains of the same species, which did not have such properties and the influence on them of two (121Sn; 137Cs) types of ionizing radiation have been studied. It was established that the most important fatty acids were hexadecanoic acid (C16:0), octadecanoic acid (C18:0), octadecenoic acid (C18:1) and octadecadienoic acid (C18:2). The strains, showing radioadaptive properties in comparison with control differed in the relative concentrations of saturated and unsaturated fatty acids and some minor components. The two types of radiation had different influence on the fatty acid profiles of the investigated strains. At dark-pigmented species C. cladosportoides, H. resinae the fatty acid unsaturation rate was higher, at strains showing radioadaptive properties (0.98) as against control ones (0.73; 0.9), and at light-pigmented species A. versicolor and P lilacinus--it was lower at strains with radioadaptive properties (1.00; 0.83) as against control ones (1.08; 0.92). The paper is presented in Ukrainian.


The effect of hemolysis on plasma antithrombin activity as determined by a chromogenic method.
The concentration of free hemoglobin (Hgb) in plasma can markedly affect the outcome of laboratory tests by interfering with the spectrophotometric absorbance of biochemical tests read at wavelengths within the absorbance range of Hgb (400-440 nm). Little is known about the effects of hemoglobinemia on antithrombin (AT) activity in human plasma samples, and we are unaware of data for canine plasma samples. The objective of this study was to determine the degree of interference by Hgb on plasma AT activity and to determine if the interference is proportional to the concentration of plasma Hgb. Two pools of test plasma, designated AT100 and AT70, were prepared. Hemolysate was prepared by washing and freeze-thawing packed red cells in a small volume of saline, followed by collection of the filtrate. Solutions of decreasing Hgb concentration were prepared and added to the test plasma pools to create a series of samples with final calculated and measured Hgb concentrations of 0, 0.5, 1.5, 2.5, 3.5, 4.5, and 5.5 g/L. AT activity, expressed as a percentage of normal human plasma, was determined using a functional chromogenic assay. Increasing concentrations of Hgb resulted in a linear decrease in AT activity. Using linear regression analysis on the 70% and 100% plasma pools, the slopes for samples containing <1.5 g Hgb/L were not significantly different from zero. Slopes for samples containing Hgb concentrations >or=1.5 g/L were significantly (P > .0001) different from zero for both plasma pools, indicating interference with the assay. The results of this study suggest it may be possible, using a conversion equation, to accurately determine AT activity in hemolyzed samples, facilitating evaluation of coagulation status in patients with intravascular hemolysis.


Variants in CDA and ABCB1 are predictors of capecitabine-related adverse reactions in colorectal cancer.
Adverse reactions to capecitabine-based chemotherapy limit full administration of cytotoxic agents. Likewise, genetic variations associated with capecitabine-related adverse reactions are associated with controversial results and a low predictive value. Thus, more evidence on the role of these variations is needed. We evaluated the association between nine polymorphisms in MTHFR, CDA, TYMS, ABCB1, and ENOSF1 and adverse reactions, dose reductions, treatment delays, and overall toxicity in 239 colorectal cancer patients treated with capecitabine-based regimens. The ABCB1*1 haplotype was associated with a high risk of delay in administration or reduction in the dose of capecitabine, diarrhea, and overall toxicity. CDA rs2072671 A was associated with a high risk of overall toxicity. TYMS rs45445694 was associated with a high risk of delay in administration or reduction in the dose of capecitabine, HFS >1 and HFS >2. Finally, ENOSF1 rs2612091 was associated with HFS >1, but was a poorer predictor than TYMS rs45445694. A score based on ABCB1-CDA polymorphisms efficiently predicts patients at high risk of severe overall toxicity (PPV, 54%; sensitivity, 43%) in colorectal cancer patients treated with regimens containing capecitabine. Polymorphisms in ABCB1, CDA, ENOSF1,and TYMS could help to predict specific and overall severe adverse reactions to capecitabine.


Different role of oxidative stress in paraventricular nucleus and rostral ventrolateral medulla in cardiovascular regulation in awake spontaneously hypertensive rats.
The rostral ventrolateral medulla (RVLM) of the brainstem and the paraventricular nucleus of the hypothalamus (PVN) are involved in the neural mechanisms of hypertension. Oxidative stress in the RVLM contributes to the enhanced central sympathetic outflow that leads to hypertension in experimental models of hypertension, such as spontaneously hypertensive rats (SHRs). We investigated the relative contribution of oxidative stress in the PVN and RVLM of SHR in blood pressure (BP) regulation. We transfected adenovirus vectors encoding the manganese superoxide dismutase gene (AdMnSOD) or β-galactosidase gene (AdLacZ) bilaterally into the RVLM or PVN. Mean arterial pressure (MAP) and heart rate (HR) were monitored using a radiotelemetry system. Oxidative stress levels in the PVN of SHR evaluated by thiobarbituric acid-reactive substances were enhanced compared with those of Wistar-Kyoto rats and reduced by MnSOD transfection compared with nontransfected SHR. MAP and HR of AdMnSOD-RVLM-transfected SHR were decreased compared with AdLacZ-RVLM-transfected SHR. In contrast, MAP of AdMnSOD-PVN-transfected SHR was not decreased compared with AdLacZ-PVN-transfected SHR, but HR was decreased compared with AdLacZ-PVN-transfected SHR. MnSOD transfection into both the RVLM and PVN of SHR decreased MAP and elicited a profound decrease in HR. These findings indicate that inhibition of oxidative stress in the PVN decreases HR, but not BP in SHR, and elicits a further decrease in HR, but not BP, by interacting with the RVLM. Taken together, the oxidative stress in the PVN and RVLM plays a different role for cardiovascular regulation in SHR.


Galectin-3 interactions with glycosphingolipids.
Galectins have essential roles in pathological states including cancer, inflammation, angiogenesis and microbial infections. Endogenous receptors include members of the lacto- and neolacto-series glycosphingolipids present on mammalian cells and contain the tetrasaccharides lacto-N-tetraose (LNT) and lacto-N-neotetraose (LNnT) that form their core structural components and also ganglio-series glycosphingolipids. We present crystallographic structures of the carbohydrate recognition domain of human galectin-3, both wild type and a mutant (K176L) that influenced ligand affinity, in complex with LNT, LNnT and acetamido ganglioside a-GM3 (α2,3-sialyllactose). Key structural features revealed include galectin-3's demonstration of a binding mode towards gangliosides distinct from that to the lacto/neolacto-glycosphingolipids, with its capacity for recognising the core β-galactoside region being challenged when the core oligosaccharide epitope of ganglio-series glycosphingolipids (GM3) is embedded within particular higher-molecular-weight glycans. The lacto- and neolacto- glycosphingolipids revealed different orientations of their terminal galactose in the galectin-3-bound LNT and LNnT structures that has significant ramifications for the capacity of galectin-3 to interact with higher-order lacto/neolacto-series glycosphingolipids such as ABH blood group antigens and the HNK-1 antigen that is common on leukocytes. LNnT also presents an important model for poly-N-acetyllactosamine-containing glycans and provides insight into galectin-3's accommodation of extended oligosaccharides such as the poly-N-acetyllactosamine-modified N- and O-glycans that, via galectin-3 interaction, facilitate progression of lung and bladder cancers, respectively. These findings provide the first atomic detail of galectin-3's interactions with the core structures of mammalian glycosphingolipids, providing information important in understanding the capacity of galectin-3 to engage with receptors identified as facilitators of major disease.


The Effects of Trivialization of Workplace Violence on Its Victims: Profession and Sex Differences in a Cross-Sectional Study among Healthcare and Law Enforcement Workers.
Workers from the law enforcement and healthcare sectors tend to normalize or mute their victimization from workplace violence (WPV). This study aims to assess the impact of the trivialization of WPV on psychological consequences for workers who have been affected by a WPV incident. The second aim is to assess the moderating effect of sex on the trivialization of WPV. The third and overarching aim is to assess the moderating effect of professional identity on the relations between individual and organizational factors and psychological consequences following a WPV incident. The findings are based on a convenience sample of 377 (204 female and 173 male) workers from the law enforcement and healthcare sectors. Individual factors (sex, age, professional identity, prior victimization, witnessing WPV, injuries, and trivialization of violence) and perceived support factors (colleagues' support and employer's support) were used as predictor variables of psychological consequences in hierarchical linear regression models. Sex was used as a moderator of trivialization while professional identity was used as a moderator of all predictors. When individual and social support factors were controlled for, normalizing violence was negatively associated with psychological consequences while perceiving a taboo associated with complaining about WPV was positively associated for all participants. When these relations were moderated by the sex of the participants and then by their professional identity, normalization was found to decrease psychological consequences only for male healthcare workers. To help employees cope with WPV, organizations should promote strategies adapted to profession and sex differences. For male healthcare workers, normalization as a cognitive coping strategy should be formally recognized. For both professions and sexes, organizational strategies that counter the perceived taboo of complaining about violence should be reinforced.


Epidural analgesia.
The most frequently used postoperative analgesia techniques are intramuscular injection (IM) and patient controlled analgesia (PCA). Recently, the use of epidural catheter injection (EPI) has been done with success. This study was done to prospectively compare these three techniques for postoperative analgesia after extensive operations upon the colon and rectum. Patients were randomized to one of three analgesia groups--IM, intramuscular morphine sulfate; PCA, patient controlled morphine sulfate, and EPI, epidural morphine sulfate. Data collected included age, time to first bowel movement, amount of narcotic, number achieving 75 per cent of preoperative forced vital capacity, postoperative pruritus, headache, nausea and vomiting, respiratory depression, atelectasis or pneumonitis. A visual analog pain scale was used to evaluate postoperative pain severity (0, no; 1, partial; 2, marked, and 3, total relief). Sixty-eight patients were eligible for study (IM, 19; PCA, 22; EPI, 23, and excluded, four). The EPI group required significantly less daily narcotic compared with either the IM or PCA groups (17.0 +/- 6.12 milligrams; 67.8 +/- 26.8 milligrams; 40.5 +/- 20.6 milligrams, respectively, less than 0.05 ANOVA) and total narcotic (81.3 +/- 31.3 milligrams; 355.4 +/- 147.7 milligrams; 215.3 +/- 105.4 milligrams, respectively, p less than 0.05 ANOVA). EPI achieves excellent pain control in more patients with a significantly lower dose of narcotics and significantly fewer pulmonary complications. Therefore, epidural analgesia is the optimal method of postoperative analgesia after extensive abdominal operations.


The aftereffects of ventriloquism: patterns of spatial generalization.
We examined how visual recalibration of apparent sound location obtained at a particular location generalizes to untrained locations. Participants pointed toward the origin of tone bursts scattered along the azimuth, before and after repeated exposure to bursts in one particular location, synchronized with point flashes of light a constant distance to their left/right. Adapter tones were presented straight ahead in Experiment 1, and in the left or right periphery in Experiment 2. With both arrangements, different generalization patterns were obtained on the visual distractor's side of the auditory adapter and onthe opposite side. On the distractor side, recalibration generalized following a descending gradient; practically no generalization was observed on the other side. This dependence of generalization patterns on the direction of the discordance imposed during adaptation has not been reported before, perhaps because the experimental designs in use did not allow its observation.


Does Weight Matter? Outcomes in Adult Patients on Venovenous Extracorporeal Membrane Oxygenation When Stratified by Obesity Class.
Many believe obesity is associated with higher rates of mortality in the critically ill. The purpose of this retrospective observational study is to evaluate the association between body mass index (BMI) and survival in patients receiving venovenous (VV) extracorporeal membrane oxygenation (ECMO) for acute hypoxic or hypercarbic respiratory failure. All of the patients admitted to a dedicated VV ECMO unit were included. Patients <18 years of age, listed for lung transplant, or underweight were excluded. ECMO outcomes, including hospital length of stay and survival to discharge, were analyzed after stratification according to BMI. Multivariate logistic and linear regression techniques were used to assess variables associated with the outcomes of death and length of stay, respectively. One hundred ninety-four patients with a median BMI of 35.7 kg/m (33-42 kg/m) were included. Obese patients were older, had higher creatinine levels, and required higher levels of positive end-expiratory pressure and mean airway pressure at time of cannulation. Survival to discharge in any group did not differ when stratified by BMI classification (P = .36). Multivariable regression did not reveal any association with greater odds of death or longer length of stay when controlling for BMI and other variables. We did not detect an association between obesity and increased mortality in patients requiring VV ECMO for acute hypoxic or hypercarbic respiratory failure. These data suggest that obesity alone should not exclude candidacy for VV ECMO. Evidence for the "obesity paradox" in this population of VV ECMO patients may be supported by these data.


Assessment of prognosis of cirrhosis.
Once patients with cirrhosis experience decompensation, early mortality risk increases sharply. Liver transplantation has transformed the prognosis of decompensated cirrhosis. Child-Pugh score has been the reference for many years for assessing the prognosis of cirrhosis. However, Child-Pugh score has important limitations among which is subjective interpretation of some of its variables, making it difficult to categorize patients according to their own disease severity. The model for end-stage liver disease (MELD) score, which was originally designed for assessing the prognosis of cirrhotic patients undergoing transjugular intrahepatic portosystemic shunt (TIPS), is a continuous score relying on three objective variables. Along with TIPS, MELD score proved to be a robust marker of early mortality across a wide spectrum of causes of cirrhosis, even though 10 to 20% of patients are still misclassified. MELD is especially useful for prioritizing candidates for transplantation according to a "sickest first" policy. However, MELD is not a universal prognostic marker of cirrhosis and several MELD exceptions require more specific approaches.


Virulence of enteropathogenic Escherichia coli, a global pathogen.
Enteropathogenic Escherichia coli (EPEC) remains an important cause of diarrheal disease worldwide. Research into EPEC is intense and provides a good virulence model of other E. coli infections as well as other pathogenic bacteria. Although the virulence mechanisms are now better understood, they are extremely complex and much remains to be learnt. The pathogenesis of EPEC depends on the formation of an ultrastructural lesion in which the bacteria make intimate contact with the host apical enterocyte membrane. The formation of this lesion is a consequence of the ability of EPEC to adhere in a localized manner to the host cell, aided by bundle-forming pili. Tyrosine phosphorylation and signal transduction events occur within the host cell at the lesion site, leading to a disruption of the host cell mechanisms and, consequently, to diarrhea. These result from the action of highly regulated EPEC secreted proteins which are released via a type III secretion system, many genes of which are located within a pathogenicity island known as the locus of enterocyte effacement. Over the last few years, dramatic increases in our knowledge of EPEC virulence have taken place. This review therefore aims to provide a broad overview of and update to the virulence aspects of EPEC.


Trends in oesophageal cancer incidence and mortality in Europe.
To monitor recent trends in mortality from oesophageal cancer in 33 European countries, we analyzed the data provided by the World Health Organization over the last 2 decades, using also joinpoint regression. For selected European cancer registration areas, we also analyzed incidence rates for different histological types. For men in the European Union (EU), age-standardized (world population) mortality rates were stable around 6/100,000 between the early 1980s and the early 1990 s, and slightly declined in the last decade (5.4/100,000 in the early 2000s, annual percent change, APC = -1.1%). In several western European countries, male rates have started to level off or decline during the last decade (APC = -3.4% in France, and -3.0% in Italy). Also in Spain and the UK, which showed upward trends in the 1990 s, the rates tended to level off in most recent years. A levelling of rates was observed only more recently in countries of central and eastern Europe, which had had substantial rises up to the late 1990 s. Oesophageal cancer mortality rates remained comparatively low in European women, and overall EU female rates were stable around 1.1-1.2/100,000 over the last 2 decades (APC = -0.1%). In northern Europe a clear upward trend was observed in the incidence of oesophageal adenocarcinoma, and in Denmark and Scotland incidence of adenocarcinoma in men is now higher than that of squamous-cell carcinoma. Squamous-cell carcinoma remained the prevalent histological type in southern Europe. Changes in smoking habits and alcohol drinking for men, and perhaps nutrition, diet and physical activity for both sexes, can partly or largely explain these trends.


The structural basis of accelerated host cell entry by SARS-CoV-2†.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of the pandemic coronavirus disease 2019 (COVID-19) that exhibits an overwhelming contagious capacity over other human coronaviruses (HCoVs). This structural snapshot describes the structural bases underlying the pandemic capacity of SARS-CoV-2 and explains its fast motion over respiratory epithelia that allow its rapid cellular entry. Based on notable viral spike (S) protein features, we propose that the flat sialic acid-binding domain at the N-terminal domain (NTD) of the S1 subunit leads to more effective first contact and interaction with the sialic acid layer over the epithelium, and this, in turn, allows faster viral 'surfing' of the epithelium and receptor scanning by SARS-CoV-2. Angiotensin-converting enzyme 2 (ACE-2) protein on the epithelial surface is the primary entry receptor for SARS-CoV-2, and protein-protein interaction assays demonstrate high-affinity binding of the spike protein (S protein) to ACE-2. To date, no high-frequency mutations were detected at the C-terminal domain of the S1 subunit in the S protein, where the receptor-binding domain (RBD) is located. Tight binding to ACE-2 by a conserved viral RBD suggests the ACE2-RBD interaction is likely optimal. Moreover, the viral S subunit contains a cleavage site for furin and other proteases, which accelerates cell entry by SARS-CoV-2. The model proposed here describes a structural basis for the accelerated host cell entry by SARS-CoV-2 relative to other HCoVs and also discusses emerging hypotheses that are likely to contribute to the development of antiviral strategies to combat the pandemic capacity of SARS-CoV-2.


Proline-rich tyrosine kinase 2 mediates gonadotropin-releasing hormone signaling to a specific extracellularly regulated kinase-sensitive transcriptional locus in the luteinizing hormone beta-subunit gene.
G protein-coupled receptor regulation of gene transcription primarily occurs through the phosphorylation of transcription factors by MAPKs. This requires transduction of an activating signal via scaffold proteins that can ultimately determine the outcome by binding signaling kinases and adapter proteins with effects on the target transcription factor and locus of activation. By investigating these mechanisms, we have elucidated how pituitary gonadotrope cells decode an input GnRH signal into coherent transcriptional output from the LH beta-subunit gene promoter. We show that GnRH activates c-Src and multiple members of the MAPK family, c-Jun NH2-terminal kinase 1/2, p38MAPK, and ERK1/2. Using dominant-negative point mutations and chemical inhibitors, we identified that calcium-dependent proline-rich tyrosine kinase 2 specifically acts as a scaffold for a focal adhesion/cytoskeleton-dependent complex comprised of c-Src, Grb2, and mSos that translocates an ERK-activating signal to the nucleus. The locus of action of ERK was specifically mapped to early growth response-1 (Egr-1) DNA binding sites within the LH beta-subunit gene proximal promoter, which was also activated by p38MAPK, but not c-Jun NH2-terminal kinase 1/2. Egr-1 was confirmed as the transcription factor target of ERK and p38MAPK by blockade of protein expression, transcriptional activity, and DNA binding. We have identified a novel GnRH-activated proline-rich tyrosine kinase 2-dependent ERK-mediated signal transduction pathway that specifically regulates Egr-1 activation of the LH beta-subunit proximal gene promoter, and thus provide insight into the molecular mechanisms required for differential regulation of gonadotropin gene expression.


[Morphology of a neonatal umbilical vein graft under prolonged experimental conditions].
The authors carried out a comparative histological and electron microscopic study of the bioprosthesis of the umbilical vein (Biograft manufactured by Meadox) and bioprosthesis obtained according to the method by M. I. Kuzin and coworkers. A far greater preservation of the new bioprosthesis was observed at long-term periods both at the histological and ultrastructural levels. Some morphological phenomena reflecting the time course of the vessel graft rearrangement were delineated 18 to 24 months after transplantation, particularly the development of "collateral" vessels in the perivenous space. The suggestion was made about an important role of Wharton's substance in the formation of the vessels.


Impact of natural light exposure on delirium burden in adult patients receiving invasive mechanical ventilation in the ICU: a prospective study.
To determine whether potential exposure to natural light via windows is associated with reduced delirium burden in critically ill patients admitted to the ICU in a single room. Prospective single-center study. Medical ICU of a university hospital, Paris, France. Adult patients receiving invasive mechanical ventilation. Consecutive patients admitted to a single room with (LIGHT group) or without (DARK group) exposure to natural light via windows were evaluated for delirium. The primary endpoint was the incidence of delirium. Main secondary endpoints included incidence of severe agitation intervened with antipsychotics and incidence of hallucinations. A total of 195 patients were included (LIGHT group: n = 110; DARK group: n = 85). The incidence of delirium was similar in the LIGHT group and the DARK group (64% vs. 71%; relative risk (RR) 0.89, 95% CI 0.73-1.09). Compared with the DARK group, patients from the LIGHT group were less likely to be intervened with antipsychotics for agitation episodes (13% vs. 25%; RR 0.52, 95% CI 0.27-0.98) and had less frequent hallucinations (11% vs. 22%; RR 0.49, 95% CI 0.24-0.98). In multivariate logistic regression analysis, natural light exposure was independently associated with a reduced risk of agitation episodes intervened with antipsychotics (adjusted odds ratio = 0.39; 95% CI 0.17-0.88). Admission to a single room with potential exposure to natural light via windows was not associated with reduced delirium burden, as compared to admission to a single room without windows. However, natural light exposure was associated with a reduced risk of agitation episodes and hallucinations.