Antivirulence activity of the human gut metabolome.
The mammalian gut contains a complex assembly of commensal microbes termed microbiota. Although much has been learned about the role of these microbes in health, the mechanisms underlying these functions are ill defined. We have recently shown that the mammalian gut contains thousands of small molecules, most of which are currently unidentified. Therefore, we hypothesized that these molecules function as chemical cues used by hosts and microbes during their interactions in health and disease. Thus, a search was initiated to identify molecules produced by the microbiota that are sensed by pathogens. We found that a secreted molecule produced by clostridia acts as a strong repressor of Salmonella virulence, obliterating expression of the Salmonella pathogenicity island 1 as well as host cell invasion. It has been known for decades that the microbiota protects its hosts from invading pathogens, and these data suggest that chemical sensing may be involved in this phenomenon. Further investigations should reveal the exact biological role of this molecule as well as its therapeutic potential. Importance: Microbes can communicate through the production and sensing of small molecules. Within the complex ecosystem formed by commensal microbes living in and on the human body, it is likely that these molecular messages are used extensively during the interactions between different microbial species as well as with host cells. Deciphering such a molecular dialect will be fundamental to our understanding of host-microbe interactions in health and disease and may prove useful for the design of new therapeutic strategies that target these mechanisms of communication.


Preparation of cultured smooth muscle cells from human myometrium for X-ray microanalysis.
Methodological aspects of the use of X-ray microanalysis in physiological and pharmacological experiments on cultured myometrial cells were investigated. Cultured human myometrial cells were grown from biopsies after detaching the fibroblasts. Of the cultured cells, 95-98% showed desmin-like immunoreactivity. Transmission electron microscopy showed that subcultured cells were different from myometrial cells in situ. The effects of washing the cells to remove external salt-rich medium were investigated. All solutions removed the external medium, resulting in lower concentrations of Na and Cl. In the cells washed with 0.3 M mannitol, most of the elemental concentrations were significantly lower than in their unwashed counterparts and those washed in the other solutions. In cells washed in either 0.15 M ammonium acetate or distilled water, no significant differences in P and K compared with their unwashed counterparts were found. There were also no significant differences between cells washed in ammonium acetate and in distilled water. In subsequent experiments ammonium acetate was used. Incubation of cells in standard Ringer's solution resulted in an increase in Na and Cl, and a decrease in K, concomitantly with an increase in Ca. Although Ringer's solution per se can elicit changes in diffusible elements in the cells, physiological and pharmacological effects of oxytocin could still be detected in Ringer's solution. However, effects of oxytocin were different when the experiment was done in culture medium, instead of in Ringer's solution.


Observations on the improvements in carbohydrate metabolism in diabetic and other morbidly obese patients after jejunoileal bypass.
In most morbidly obese patients with diabetes, fasting plasma glucose levels decreased immediately after jejunoileal bypass operations, with patients often becoming normoglycemic before discharge from the hospital. All 12 patients who had required insulin or orally administered hypoglycemic agents preoperatively were able to discontinue the medication shortly after the operation. Oral glucose tolerance test curves in all morbidly obese patients had a flattened pattern, in the normal range, postoperatively. Serum insulin levels, which had been elevated preoperatively, decreased significantly, both in fasting and postglucose determinations. Results of intravenous glucose tolerance tests showed little change in the early period after operation. Improvement in carbohydrate metabolism may be due to several factors and does not appear to be dependent upon massive weight loss. Major factors may include decreased absorption of carbohydrates and amino acids, decreased oral caloric intake, increased insulin sensitivity and decreased output of gastric inhibitory polypeptide as well as the eventual weight loss.


Dealcoholized red wine containing known amounts of resveratrol suppresses atherosclerosis in hypercholesterolemic rabbits without affecting plasma lipid levels.
Moderate consumption of red wine is associated with a reduced risk of coronary heart disease (CHD). This phenomenon is based on data from epidemiological observations known as the French paradox, and has been attributed to CHD-protective phytochemicals, e.g. resveratrol in red wine. Since red wine also contains alcohol, it is conceivable that alcohol interacts with resveratrol to elicit the observed cardioprotective effects. To determine whether resveratrol has alcohol-independent affects, we compared cardioprotective properties of dealcoholized Chinese red wine with alcohol-containing Chinese red wine having comparable amounts of resveratrol, using a hypercholesterolemic rabbit model and resveratrol as a reference. Animals fed a high cholesterol (1.5%) diet were simultaneously given water containing resveratrol (3 mg/kg/day) or red wine (4 ml/kg/day) containing 3.98 mg/l and 3.23 mg/l resveratrol for regular and dealcoholized red wine, respectively, for a 12-week duration. Total, HDL- and LDL-cholesterol and triglyceride levels in the plasma were measured before and after the cholesterol challenge. Atherosclerotic plaques in the thoracic aorta were evaluated using histochemical methods. Vascular and endothelial functions in the femoral artery were also assessed by ultrasonographic image analysis. High cholesterol-fed animals showed a significant increase in plasma levels of total, HDL- and LDL-cholesterol, but not triglycerides, compared to those fed a regular diet. Dietary cholesterol-elicited lipid changes were similarly observed in animals concurrently fed dealcoholized red wine, red wine or resveratrol. In contrast, whereas atherosclerotic lesions were clearly evident in specimens prepared from the thoracic aorta of high cholesterol-fed animals, the size, density, and mean area of atherosclerotic plaques, and thickness of the intima layer were significantly reduced in rabbits given dealcoholized red wine, red wine, or resveratrol. These results were in agreement with data obtained by an ultrasound analysis of endothelial function, which showed a 25% reduction in flow-mediated dilation (FMD) in rabbits fed a high cholesterol diet compared to animals on control diet. This decrease was effectively prevented by the simultaneous exposure to dealcoholized red wine, red wine, or resveratrol. Our study shows that animals given dealcoholized red wine exhibited cardio-active effects comparable to those of animals orally administered resveratrol, and suggests that wine polyphenolics, rather than alcohol present in red wine, suffice in exerting cardioprotective properties. The results also provide support for the notion that resveratrol and phytochemicals in red wine can suppress atherosclerosis without affecting plasma lipid levels.


[Natural and synthetic inhibitors of thrombin. I. Natural inhibitors].
Structures and properties of main physiological (heparin, antithrombin III, heparin cofactor II) and nonphysiological (hirudin, thrombin-binding aptamers, cyclotheonamides) natural thrombin inhibitors and its fragments and synthetic analogs (hirugen, hirulogs, hirunorms, pentasaccharides, macrocyclic alpha-keto amides) were reviewed. The molecular mechanisms of interaction of these compounds with thrombin and their anticoagulant activity at preclinical and clinical trials are discussed. The examined of natural thrombin inhibitors and their synthetic fragments and analogs are perspective for prophylaxis and treatment of different thrombo-embolic diseases.


Choice of Endoscopic Procedure in Children With Clinically Suspected Gastrointestinal Graft-versus-host Disease.
Gastrointestinal graft-versus-host disease (GI-GVHD) is a potentially life-threatening complication after hematopoietic stem cell transplantation. Symptoms indicating GI-GVHD motivates endoscopy with biopsy sampling and histopathological confirmation. Optimal extent of endoscopy in children is, however, presently unknown. Therefore, we aimed to evaluate whether biopsies from the rectosigmoid area versus the rest of the colon/ileocolon with or without biopsies from simultaneous upper endoscopy, were equally reliable for detection of GI-GVHD and relevant differential diagnoses. Retrospective multicenter study based on histopathological re-evaluation of biopsies and hospital record data, collected from children with suspected GI-GVHD. Forty-four children with 51 endoscopic occasions (81 procedures) were included. Thirty-nine of 51 (76.5%) were diagnosed as GI-GVHD, 14 (27.4%) received a differential diagnosis and 7 (13.7%) had normal histology findings. Comorbidity, that is, simultaneous detection of a differential diagnosis and GI-GVHD, was observed in 9 (23.1%) cases. Cytomegalovirus infection was the most frequent differential diagnosis, 6 of 7 were detected in biopsies from rectosigmoid and esophagogastroduodenal areas. Sensitivity for detection of GI-GVHD in biopsies collected from rectosigmoid-ileocolonic-, rectosigmoid-, or esophagogastroduodenal areas were 97.4%, 84.6%, 83.3%, respectively, and 97.4% when the latter 2 were merged. The difference, nondetected GI-GVHD in the rectosigmoid area versus detected elsewhere in the GI tract, was statistically significant (P = 0.03). Biopsies collected from the rectosigmoid area solely were not optimal for detection of pediatric GI-GVHD. When biopsy sampling from rectosigmoid and upper GI tract areas was combined, the sensitivity for GI-GVHD was, however, equally high as for ileocolonoscopy or full upper and lower endoscopy.


Conceptualizing evolutionary novelty: moving beyond definitional debates.
According to many biologists, explaining the evolution of morphological novelty and behavioral innovation are central endeavors in contemporary evolutionary biology. These endeavors are inherently multidisciplinary but also have involved a high degree of controversy. One key source of controversy is the definitional diversity associated with the concept of evolutionary novelty, which can lead to contradictory claims (a novel trait according to one definition is not a novel trait according to another). We argue that this diversity should be interpreted in light of a different epistemic role played by the concept of evolutionary novelty-the structuring of a problem space or setting of an explanatory agenda-rather than the concept's capacity to categorize traits as novel. This distinctive role is consistent with the definitional diversity and shows that the concept of novelty benefits ongoing investigation by focusing attention on answering different questions related to comprehending the origins of novelty. A review of recent theoretical and empirical work on evolutionary novelty confirms this interpretation.


The current status of biomarkers for the diagnosis of nosocomial pneumonias.
Nosocomial pneumonia is a frequent and severe nosocomial infection divided in two distinct groups: hospital-acquired pneumonia and ventilator-associated pneumonia (VAP). In this context, the VAP is notoriously difficult to diagnose clinically, resulting from the lack of a 'gold standard' method of diagnosis. The use of biomarkers may potentially improve the early diagnosis of infections allowing earlier and better identification and treatment. An exhausting list of biomarkers has been studied and although far from perfect, procalcitonin (PCT) and C-reactive protein (CRP) are the most studied biomarkers used in clinical practice. Data coming from literature suggests the use of PCT for VAP prognosis and as a based algorithm tool for the reduction of duration of pneumonia therapy, as well as, the use of the CRP dynamics to the early prediction of VAP and the response to the antibiotics. The evidence for the use of biomarkers to diagnose nosocomial pneumonia as a stand-alone tool is low to moderate. Improved performance for both PCT and CRP can be obtained by using them in association with clinical features or scoring systems but prospective studies are still needed to validate this hypothesis.


Bio-inspired nano-engineering of an ultrahigh loading 3D hierarchical Ni@NiCo2S4/Ni3S2 electrode for high energy density supercapacitors.
Energy density has become a critical barrier in supercapacitor engineering and improvement of the electrode-loading is urgently demanded. However, there is conflict between the high loading and good electrochemical properties of supercapacitors. Herein, ultrahigh loading (10.33 mg·cm-2) 3D hierarchical NiCo2S4/Ni3S2 on Ni foam with outstanding performance is obtained via bio-inspired nano-engineering, which contains compact nanowire arrays catching urchin-like micro-particles. Using this high-loading material as a binder-free electrode achieves excellent areal capacitances with 16.90 F·cm-2 at 10.33 mA·cm-2 and 1.17 F·cm-2 at 5.17 mA·cm-2 in a three-electrode system and asymmetric supercapacitor device, respectively. The device also exhibits a high energy density of 4.69 W h m-2 (power density of 10.33 W·m-2) and an outstanding stability of 91.4% after 8000 cycles (20.66 mA·cm-2). Its excellent performance is attributed to the well-designed structure and composition: (i) a large contact area with the electrolyte raises the utilization efficiency of the active material, therefore guaranteeing the high capacitance of the active materials; (ii) the high electronic conductivity network constructed through NiCo2S4 and the short diffusion length boost its rate performance; (iii) the reserved space in the hierarchical structure could hold the volume change and enhance the cycling performance of the electrode in the charge/discharge cycles. Thus, this work not only provides a method for the construction of a high-loading and high-performance electrode for asymmetric supercapacitors, but could also shed light on the design of compact nano-materials for other energy storage systems.


An integrin-targeting nanosystem as a carrier of the selenadiazole derivative to induce ROS-mediated apoptosis in bladder cancer cells, from rational design to action mechanisms.
The development of novel therapeutics for patients with bladder cancer is an important area of research, particularly considering the rather limited treatment options currently available. In this study, we designed and synthesized a conjugate of cancer-targeting selenadiazole derivative BSeC (benzo[1,2,5]selenadiazole-5-carboxylic acid) and the RGD (arginine-glycine-aspartate) peptide, which was used as a targeting molecule, using a PEI polymer as a linker. The results showed that BSeC-PEI-RGD formed core-shell spherical nanoparticles with improved stability in physiological and low pH solutions. The cancer-targeting design significantly enhanced cellular uptake of BSeC-PEI-RGD and decreased its cytotoxicity to normal cells. The nanoparticles could inhibit the migration and invasion of EJ and T24 bladder cancer cell and reduce cancer cell proliferation through the induction of reactive oxygen species (ROS)-dependent apoptosis and mitochondrial dysfunction. Further mechanistic studies using western blotting showed that BSeC-PEI-RGD triggered bladder cancer cell apoptosis by activating p38, JNK and p53 and by inactivating AKT and ERK. In summary, this study demonstrates the rational design of a polymer-based cancer-targeting nanosystem as a carrier of the selenadiazole derivative to treat bladder cancer.


Dopaminergic and non-dopaminergic pharmacological hypotheses for gait disorders in Parkinson's disease.
Gait disorders form one component of the axial disorders observed in Parkinson's disease (PD). Indeed, short steps with a forward-leaning stance are diagnostic criteria for PD in the early stages of the condition. Gait disorders also represent a major source of therapeutic failure in the advanced stages of PD (with the appearance of freezing of gait and falls) because they do not respond optimally to the two hand late-stage therapeutics--levodopa and electrical subthalamic nucleus (STN) stimulation. The late onset of doparesistance in these disorders may be linked to propagation of neurodegeneration to structures directly involved in gait control and to non-dopaminergic neurotransmitter systems. The coeruleus locus (a source of noradrenaline) is rapidly and severely affected, leading to a major motor impact. The pedunculopontine nucleus (PPN) and lateral pontine tegmentum (rich in acetylcholine) are both involved in gait. Degenerative damage to the serotoninergic raphe nuclei appears to be less severe, although serotonin-dopamine interactions are numerous and complex. Lastly, dopaminergic depletion leads to glutamatergic hyperactivity of the efferent pathways from the the STN to the PPN. However, the relationships between the various parkinsonian symptoms (and particularly gait disorders) and these pharmacological targets have yet to be fully elucidated. The goal of this review is to develop the various pathophysiological hypotheses published to date, in order to underpin and justify ongoing fundamental research and clinical trials in this disease area.


Feasibility of minimally invasive sacropelvic fixation: percutaneous S2 alar iliac fixation.
A cadaveric study with postoperative computed tomography scan to evaluate instrumentation placement. To successfully place percutaneous sacropelvic instrumentation. S2 iliac fixation has been in use clinically at some centers. Recently, anatomic data have been presented on the technique. The purpose of this study is to determine the feasibility of percutaneous placement of S2 iliac sacropelvic fixation (1) without damage to vital structures and (2) with in-line placement with S1 pedicle screws. Eight cadaveric spines were used in this study. Percutaneous pedicle screws were placed from L3-S1 in 4 and from L2-S1 in the remainder. Percutaneous S2 iliac screws were placed using a modification of the open technique. Rods were placed using minimally invasive techniques. All specimens were CT scanned. Trajectory of the screws was measured from CT scans. Maximal length was judged by a k-wire left in the S2 iliac screw. CT scans were critically evaluated for risks to visceral and neurovascular structures as well as cortical breaches. Average length of the screws was 92.5 mm (range, 69-120 mm). No screw was intrapelvic or risked any visceral or neurovascular structure. No screws violated the cortex of the ilium. All S2 iliac screws were in-line with the S1 pedicle screws. The average cephalocaudad trajectory was 29 degrees caudal from direct lateral. The average anterior-posterior angulation was 42 degrees from a horizontal line connecting the PSIS. Use of the S2 iliac technique may be a viable option in minimally invasive thoracolumbar deformity surgery. The screws were all in-line and connected easily to the cephalad instrumentation. On average, a length of approximately 90 mm was attained. No visceral or neurovascular structure was injured. Visualization of the first dorsal foramen and a standard anteroposterior and inlet radiograph were used for placement.


Eastern equine encephalitis.
We describe a patient who died from a fulminant presentation of encephalitis. After an exhaustive search, we found no treatable cause. Postmortem PCR analysis of brain tissue led to a diagnosis of eastern equine encephalitis. We have identified several clinical pearls that may assist others in making the diagnosis earlier in the disease course.


Cognition commands change.
Problematic behavior can be viewed as the result of the interaction between parents and children as guided by operating assumptions. Preliminary research suggests that enduring and generalized adaptive changes in problematic behavior are engendered when behavior modification techniques like praise are augmented with cognitive techniques like self talk. The changes may be more enduring and generalized when these techniques are used not only with children but also with and by parents. However, research is needed to validate this working hypothesis as well as to determine the most effective behaviors and cognitions of parents necessary to bring about and maintain change. Until this research has been validated, clinicians may utilize these techniques both with and by parents and with children on the bases of face validity and of extending logically the findings in the areas of behavior modification and cognitive-behavioral techniques.


Evaluation of the role of proline residues flanking the RGD motif of dendroaspin, an inhibitior of platelet aggregation and cell adhesion.
The effect of a panel of proline mutants of dendroaspin, an inhibitor of platelet aggregation and cell adhesion, including A(42)-dendroaspin, A(47)-dendroaspin, A(49)-dendroaspin, A(42,47)-dendroaspin and A(47,49)-dendroaspin, was investigated using platelet-aggregation and cell-adhesion assays. Here we show that a single alanine-for-proline substitution did not affect potency when measured as the ability either to inhibit platelet aggregation induced by ADP (IC(50) approximately 170 nM) or to block transfected A375-SM cell adhesion to fibrinogen in the presence of Mn(2+) as compared with wild-type dendroaspin. By comparison, double proline substitution with alanines significantly reduced the potency in both assays by approx. 5-8-fold. These observations, therefore, suggest that proline residues flanking the RGD motif in dendroaspin and other RGD-containing venom proteins, e.g. disintegrins, may contribute to maintaining a favourable conformation for the solvent-exposed RGD site for its recognition by integrin receptors.


International exchanges in primary care-learning from thy neighbour.
This systematic review describes how international exchange programmes in primary care have been received and evaluated. Electronic databases (MEDLINE, Embase, PsycINFO, EBM reviews, CAB abstracts and PubMED) were searched to identify articles where the main focus of the study was exchanges undertaken in primary care/family medicine until March 2016. Articles were included if they (i) discussed participant exchanges in primary care; (ii) presented associated outcome data-this included (a) individual/group experience of exchange; (b) mechanism of exchange and (c) observations during the exchange. A narrative synthesis was performed of the heterogeneous data identified. Twenty-nine studies were included. Exchange locations varied across the world with the largest number in Europe. Participants came from a range of backgrounds including medical students, nurses, General Practitioners (GP), GP trainees (GPTs) and visiting scholars/professors. Exchange duration ranged from 3 days to 2 years. Key themes were identified from analysis of the studies with illustrative quotes from the included studies provided. Four key areas were discussed in relation to exchange experience: learning opportunities and new knowledge; comparative observation; knowledge gained and translational learning. Primary care international exchanges provide a rich source of cross-country learning. This review identified that exchange participants benefit both personally and professionally, equipping them with translatable skills to improve the care provided to their patients.


Comparing the performance-enhancing effects of squats on a vibration platform with conventional squats in recreationally resistance-trained men.
The purpose of this investigation was to compare the performance-enhancing effects of squats on a vibration platform with conventional squats in recreationally resistance-trained men. The subjects were 14 recreationally resistance-trained men (age, 21-40 years) and the intervention period consisted of 5 weeks. After the initial testing, subjects were randomly assigned to either the "squat whole body vibration" (SWBV) group (n = 7), which performed squats on a vibration platform on a Smith Machine, or the "squat"(S) group (n = 7), which performed conventional squats with no vibrations on a Smith Machine. Testing was performed at the beginning and the end of the study and consisted of 1 repetition maximum (1RM) in squat and maximum jump height in countermovement jump (CMJ). A modified daily undulating periodization program was used during the intervention period in both groups. Both groups trained at the same percentage of 1RM in squats (6-10RM). After the intervention, CMJ performance increased significantly only in the SWBV (p < 0.01), but there was no significant difference between groups in relative jump height increase (p = 0.088). Both groups showed significant increases in 1RM performance in squats (p < 0.01). Although there was a trend toward a greater relative strength increase in the SWBV group, it did not reach a significant level. In conclusion, the preliminary results of this study point toward a tendency of superiority of squats performed on a vibration platform compared with squats without vibrations regarding maximal strength and explosive power as long as the external load is similar in recreationally resistance-trained men.


Chiral discrimination of α-hydroxy acids and N-Ts-α-amino acids induced by tetraaza macrocyclic chiral solvating agents by using 1H NMR spectroscopy.
In the field of chiral recognition, reported chiral discrimination by 1H NMR spectroscopy has mainly focused on various chiral analytes with a single chiral center, regarded as standard chiral substrates to evaluate the chiral discriminating abilities of a chiral auxiliary. Among them, chiral α-hydroxy acids, α-amino acids and their derivatives are chiral organic molecules involved in a wide variety of biological processes, and also play an important role in the area of preparation of pharmaceuticals, as they are part of the synthetic process in the production of chiral drug intermediates and protein-based drugs. In this paper, several α-hydroxy acids and N-Ts-α-amino acids were used to evaluate the chiral discriminating abilities of tetraaza macrocyclic chiral solvating agents (TAMCSAs) 1a-1d by 1H NMR spectroscopy. The results indicate that α-hydroxy acids and N-Ts-α-amino acids were successfully discriminated in the presence of TAMCSAs 1a-1d by 1H NMR spectroscopy in most cases. The enantiomers of the α-hydroxy acids and N-Ts-α-amino acids were assigned based on the change of integration of the 1H NMR signals of the corresponding protons. The enantiomeric excesses (ee) of N-Ts-α-amino acids 11 with different optical compositions were calculated based on the integration of the 1H NMR signals of the CH3 protons (Ts group) of the enantiomers of (R)- and (S)-11 in the presence of TAMCSA 1b. At the same time, the possible chiral discriminating behaviors have been discussed by means of the Job plots of (±)-2 with TAMCSAs 1b and proposed theoretical models of the enantiomers of 2 and 6 with TAMCSA 1a, respectively.


Human umbilical cord blood cells can be induced to express markers for neurons and glia.
Rare cells are present in human umbilical cord blood that do not express the hematopoietic marker CD45 and in culture do not produce cells of hematopoietic lineage. These umbilical cord multipotent stem cells (UC-MC) behave as multilineage progenitor cells (stem cells) and can be expanded in tissue culture. Exposure to basic fibroblast growth factor (bFGF) and human epidermal growth factor (hEGF) for a minimum of 7 days in culture induces expression of neural and glial markers. Western immunoblots demonstrate expression of both beta-tubulin III and glial fibrillary acidic protein (GFAP). Immunocytochemistry of the cells showed intense labeling to both compounds on the intracellular cytoskeleton. The oligodendrocyte cell surface marker galactocerebroside (Gal-C) was present on most cells. Many cells show dual labeling, expressing both neuronal and glial markers.


Diversity of the lichenized fungi in King George Island, Antarctica, revealed by phylogenetic analysis of partial large subunit rDNA sequences.
Lichens are predominant and important components of flora in the terrestrial ecosystem of Antarctica. However, relatively few researches on the phylogenetic position of Antarctic lichen-forming fungi have been accomplished. In this study, partial sequences of nuclear large subunit rDNAs from 50 Antarctic specimens were obtained and the phylogeny was reconstructed. Antarctic lichen species were distributed among 4 orders, including the monophyletic order Agyrales, paraphyletic orders Pertusariales and Teloschistales, and polyphyletic order Lecanorales. Species diversity was highest in the order Lecanorales, followed by Teloschistales and Pertusariales. Based on the phylogeny and sequence similarity analyses, it is proposed that the taxonomy of Stereocaulon alpinum, Physcia caesia, Usnea aurantiacoatra, and Cladonia species should be revised by careful examination of their phenotypic and molecular characteristics. Six species known to be endemic to Antarctica, Catillaria corymbosa, Himantormia lugubris, Leptogium puberulum, Pertusaria pertusa, Rhizoplaca aspidophora, and Umbilicaria antarctica, formed unique lineages, implying independent origins in the Antarctic area.


Sustained release of matrix metalloproteinase-3 to trabecular meshwork cells using biodegradable PLGA microparticles.
Accumulation of extracellular matrix (ECM) materials in the trabecular meshwork (TM) is believed to be a contributing factor to intraocular pressure (IOP) elevation, a risk factor/cause of primary open angle glaucoma, a major blinding disease. Matrix metalloproteinase-3 (MMP-3) is one of the proteinases that can effectively degrade ECM elements such as fibronectin, and MMP-3 delivery to the TM represents a promising approach for IOP reduction and treatment of glaucoma. In this study, we tested the feasibility of using polymeric microparticles to achieve a slow and sustained release of active MMP-3 to cultured human TM cells. β-Casein, with molecular weight (24 kDa) and hydrophobicity similar to those of the active MMP-3 fragment (19.2 kDa), was first employed as a model for initial testing. β-casein was encapsulated into poly(lactic-co-glycolic acid) (PLGA) microparticles using a double emulsion procedure at an encapsulation efficiency of approximately 45%. The PLGA microparticles were chosen given their biocompatibility and the proven capacity of sustained release of encapsulated molecules. The release test conducted in the culture medium showed a slow and sustained release of the protein over 20 days without a significant initial burst release. Active MMP-3 was subsequently encapsulated into PLGA microparticles with an encapsulation efficiency of approximately 50%. A biofunctional assay utilizing human TM cells was set up in which the reduction of fibronectin was used as an indicator of enzyme activity. It was observed that fibronectin staining was markedly reduced by the medium collected from MMP-3-microparticle-treated cultures compared to that from blank- and β-casein-microparticle controls, which was validated using a direct MMP-3 activity assay. The controlled release of MMP-3 from the microparticles resulted in sustained degradation of fibronectin up to 10 days. This proof-of-concept undertaking represents the first study on the controlled and sustained release of active MMP-3 to TM cells via encapsulation into PLGA microparticles as a potential treatment of glaucoma.


Inhaling and lung cancer: an anomaly explained.
An objective index of inhaling cigarette smoke based on carboxyhaemoglobin concentrations and the carbon monoxide yields of cigarettes was used to investigate possible systematic differences in the extent of inhaling among light and heavy smokers when classified according to their self described inhaling habits. A total of 2108 men who smoked cigarettes were studied. Heavy smokers (20 or more cigarettes a day) had a higher average inhaling index than light smokers (fewer than 20 cigarettes a day) both among those who said that they inhaled and among those who said that they did not. This observation, together with indirect evidence that heavy smokers who inhale deeply may to some extent avoid depositing smoke condensate on their main bronchial epithelium, explains a hitherto unresolved anomaly--namely, that the risk of lung cancer is less among heavy cigarette smokers who say that they inhale than it is among those who say that they do not inhale.


A y(+)LAT-1 mutant protein interferes with y(+)LAT-2 activity: implications for the molecular pathogenesis of lysinuric protein intolerance.
Lysinuric protein intolerance (LPI) is an inherited aminoaciduria caused by defective cationic amino acid (CAA) transport at the basolateral membrane of epithelial cells in the intestine and kidney. The SLC7A7 gene, mutated in LPI, encodes the y(+)LAT-1 protein, which is the light subunit of the heterodimeric CAA transporter in which 4F2hc is the heavy chain subunit. Co-expression of 4F2hc and y(+)LAT-1 induces the y(+)L activity. This activity is also exerted by another complex composed of 4F2hc and y(+)LAT-2, the latter encoded by the SLC7A6 gene and more ubiquitously expressed than SLC7A7. On the basis of both the pattern of expression and the transport activity, y(+)LAT-2 might compensate for CAA transport when y(+)LAT-1 is defective. By expression in Xenopus laevis oocytes and mammalian cells, we functionally analysed two SLC7A7 mutants, E36del and F152L, respectively, the former displaying a partial dominant-negative effect. The results of the present study provide further insight into the molecular pathogenesis of LPI: a putative multiheteromeric structure of both [4F2hc/y(+)LAT-1] and [4F2hc/y(+)LAT-2], and the interference between y(+)LAT-1 and y(+)LAT-2 proteins. This interference can explain why the compensatory mechanism, that is, an increased expression of SLC7A6 as seen in lymphoblasts from LPI patients, may not be sufficient to restore the y(+)L system activity.


Adnexal masses in patients with colorectal cancer.
To evaluate the clinicopathological features and outcomes of patients undergone surgery for benign and metastatic adnexal masses during the surveillance of colorectal cancer (CRC). A single institute retrospective study involving 90 patients diagnosed with CRC that undergone surgery at Hacettepe University Hospital between 2000-2018. Clinicopathological and survival data was obtained from hospital records. Elevated blood levels of CEA (HR, 1.23; 95 % CI: 1.03-1.24; p = 0.01), adnexal masses larger than 5 cm (HR, 3.296; 95 % CI: 1.527-7.076; p = 0.002), bilaterality of adnexal mass (HR, 2.200; 95 % CI: 2.464-11.969; p = 0.001) and high PCI score (HR, 0.150; 95 % CI: 0.044-0.479; p = 0.01) were found to be significantly associated with ovarian metastasis. There was a significant difference in overall survival (OS), with respect to complete and incomplete resection in adnexal surgery of ovarian metastasis (46.6 vs. 29.6 months; p = 0.004). The median survival time was 32.8 months for patients with ≤ 24 months interval time to adnexal metastasis surgery, and 48.5 months for patients with >24 months interval time to adnexal metastasis surgery (p = 0.001). This study showed that numerous clinicopathological variables such as bilaterality and size of adnexal mass, serum blood levels of CEA and PCI score may have a significant impact on the prediction and management of ovarian masses diagnosed during CRC surveillance. Complete resection and interval time to adnexal surgery is significantly associated with OS.


Organizing organ donation.
Organ donation for transplantation involves intensive care staff in extra time and work. However, the results of renal, cardiac and hepatic transplantation are now so good (over 80% success) that every effort should be made to refer all suitable organ donors to the local transplant unit.


Climate warming and the decline of Taxus airborne pollen in urban pollen rain (Emilia Romagna, northern Italy).
Woody plant performance in a changing global environment has always been at the centre of palaeoenvironmental and long-term climate reconstructions carried out by means of pollen analysis. In Mediterranean regions, Taxus constitutes the highest percentage in past pollen diagrams from cold or cool periods, and therefore it is generally considered a good index to infer climate features from past records. However, a comparison of these inferences with the true current trends in pollen production has not been attemped until now. This study reports the decline of airborne pollen of Taxus observed in Emilia Romagna, a region of northern Italy, during the period 1990-2007. Phenological observations on four male specimens and microscopic examination of fresh pollen were made in order to check Taxus flowering time and pollen morphology. Airborne pollen was monitored through continuous sampling with a Hirst volumetric sampler. In the 18-year long period of investigation, Taxus pollen production has decreased, while total woody pollen abundance in air has increased. The trend of the Taxus pollen season shows a delay at the beginning, a shortening of the pollen period, and an advance of the end of the pollen season. This was interpreted as a response to climate warming. In particular, Taxus follows the behaviour of winter-flowering plants, and therefore earlier pollination is favoured at low autumn temperatures, while late pollination occurs more often, most likely after warm autumn temperatures.


Single and combined lesions of the cats thalamic mediodorsal nucleus and the mamillary bodies lead to severe deficits in the acquisition of an alternation task.
The acquisition of a spatial alternation task was tested in five groups of adult cats. Two groups were used for control; one of them consisted of nonlesioned cats, the other of cats with chemical lesions (ibotenic acid) of the ventral tegmental area. The other three groups of cats received chemical lesions of the mediodorsal nucleus of the thalamus, the mamillary bodies, or of both structures together. Cats with lesions of the ventral tegmental area were non-significantly impaired in comparison to non-lesioned cats in the acquisition of the task. Cats with lesions of the mediodorsal nucleus, the mamillary bodies, or with combined lesions of both structures were significantly impaired, compared to the two control groups; among each other, however, their performance rates were similar. As none of the cats with single or combined lesions of the mamillary and the mediodorsal nuclei manifested observable abnormalities with respect to motor, motivational, or emotional behavior during their testing period, it is concluded that their--compared to the other two groups--inferior performance was due to a memory defect. As all three kinds of lesions resulted in a similar defect, it is suggested that the mamillary and the mediodorsal nuclei transmit information to one or more common target area(s), which is (are) dependent on information from either nucleus to operate effectively.


Primary liver cancer in the Transvaal.
The clinical picture of 60 cases of primary liver cancer is reported. Distended abdominal veins found in a substantial number of cases, often point to collateral circulation due to a membrane in the inferior vena cava (budd-Chiari syndrome). Alpha-foetoprotein, alkaline phosphatase and gamma-glutamyltranspeptidase were the only reliable laboratory pointers to the disease. Ultrasound study proved to be very helpful. Thrombocytopenia was found in 5 out of 12 of our patients.


Cigarette smoke extract promotes proliferation of airway smooth muscle cells in asthmatic rats via regulating cyclin D1 expression.
Increased proliferation of airway smooth muscle cells (ASMCs) are observed in asthmatic patients and smoking can accelerate proliferation of ASMCs in asthma. To elucidate the molecular mechanisms leading to these changes, we studied in vitro the effect of cigarette smoke extract (CSE) on the proliferation of ASMCs and the expression of cyclin D1, an important regulatory protein implicated in cell cycle. ASMCs cultured from 8 asthmatic Brown Norway rats were studied. Cells between passage 3 and 6 were used in the study and were divided into control group, pcDNA3.1 group, pcDNA3.1-antisense cyclin D1 (ascyclin D1) group, CSE group, CSE + pcDNA3.1 group and CSE + pcDNA3.1-ascyclin D1 group based on the conditions for intervention. The proliferation of ASMCs was examined with cell cycle analysis, MTT colorimetric assay and proliferating cell nuclear antigen (PCNA) immunocytochemical staining. The expression of cyclin D1 was detected by reverse transcriptase-PCR (RT-PCR) and Western blotting. (1) The percentage of S + G2M phase, absorbance value at 490 nm wavelength (A(490)) and the expression rate of PCNA protein in CSE group were (31.22 +/- 1.17)%, 0.782 +/- 0.221, (90.2 +/- 7.0)% respectively, which were significantly increased compared with those of control group ((18.36 +/- 1.02)%, 0.521 +/- 0.109, and (54.1 +/- 3.5)%, respectively) (P < 0.01). After the transfection with antisense cyclin D1 plasmid for 30 hours, the percentage of S + G2M phase, A(490) and the expression rate of PCNA protein in ASMCs were much lower than in untreated cells (P < 0.01). (2) The ratios of A(490) of cyclin D1 mRNA in CSE group was 0.288 +/- 0.034, which was significantly increased compared with that of control group (0.158 +/- 0.006) (P < 0.01). After the transfection with antisense cyclin D1 plasmid for 30 hours, the ratios of A(490) of cyclin D1 mRNA in ASMCs was much lower than in untreated cells (P < 0.01). (3) The ratios of A(490) of cyclin D1 protein expression in CSE group was 0.375 +/- 0.008, which was significantly increased compared with that of control group (0.268 +/- 0.004) (P < 0.01). After the transfection with antisense cyclin D1 plasmid for 30 hours, the ratios of A(490) of cyclin D1 protein expression in ASMCs was much lower than in untreated cells (P < 0.01). CSE may increase the proliferation of ASMCs in asthmatic rats via regulating cyclin D1 expression.


Preserved insulin sensitivity and {beta}-cell activity, but decreased glucose effectiveness in normal-weight women with the polycystic ovary syndrome.
Insulin resistance and hyperinsulinemia are often considered intrinsic features of the polycystic ovary syndrome (PCOS). Nevertheless, conflicting results of insulin sensitivity and secretion have been obtained in the subgroup of normal-weight women with PCOS. Differences in body composition, ethnicity, and diet composition and a family history of metabolic diseases may act as confounding variables in women with PCOS. In the present study, insulin sensitivity and secretion were estimated by an iv glucose tolerance test (IVGTT), analyzed by minimal models, in 20 normal-weight healthy women with PCOS and no family history of type 2 diabetes mellitus and in 20 normally ovulating women, matched for age and body mass index. Insulin sensitivity [mean (95% confidence intervals); PCOS 4.0 (2.8-5.1) vs. controls 4.5 (3.5-5.4) 10(-4) min(-1)/microU.ml], and insulin secretion, expressed as the acute insulin response to glucose [PCOS 3.7 (3.3-4.2) vs. controls 3.7 (3.4-4.0) microU/ml] were similar in the two groups. The women with PCOS showed an increased proportion of total body fat (PCOS 29% vs. controls 27.2%; P < 0.01). They also showed decreased glucose effectiveness, i.e. the proportion of glucose uptake independent from insulin activity [PCOS 2.6 (2.1-3.0) vs. controls 3.8 (3.0-4.6) mg x 100 min(-1); P = 0.01]. The levels of insulin sensitivity and of glucose effectiveness did not correlate in either group. Whether the isolated finding of decreased glucose effectiveness could reflect an early stage in the development of the metabolic aberrations often associated with the syndrome remains to be clarified.


The missing indicator approach for censored covariates subject to limit of detection in logistic regression models.
In several biomedical studies, one or more exposures of interest may be subject to nonrandom missingness because of the failure of the measurement assay at levels below its limit of detection. This issue is commonly encountered in studies of the metabolome using tandem mass spectrometry-based technologies. Owing to a large number of metabolites measured in these studies, preserving statistical power is of utmost interest. In this article, we evaluate the small sample properties of the missing indicator approach in logistic and conditional logistic regression models. For nested case-control or matched case control study designs, we evaluate the bias, power, and type I error associated with the missing indicator method using simulation. We compare the missing indicator approach to complete case analysis and several imputation approaches. We show that under a variety of settings, the missing indicator approach outperforms complete case analysis and other imputation approaches with regard to bias, mean squared error, and power. For nested case-control and matched study designs of modest sample sizes, the missing indicator model minimizes loss of information and thus provides an attractive alternative to the oft-used complete case analysis and other imputation approaches.


A proteomic view at the biochemistry of syntrophic butyrate oxidation in Syntrophomonas wolfei.
In syntrophic conversion of butyrate to methane and CO2, butyrate is oxidized to acetate by secondary fermenting bacteria such as Syntrophomonas wolfei in close cooperation with methanogenic partner organisms, e.g., Methanospirillum hungatei. This process involves an energetically unfavourable shift of electrons from the level of butyryl-CoA oxidation to the substantially lower redox potential of proton and/or CO2 reduction, in order to transfer these electrons to the methanogenic partner via hydrogen and/or formate. In the present study, all prominent membrane-bound and soluble proteins expressed in S. wolfei specifically during syntrophic growth with butyrate, in comparison to pure-culture growth with crotonate, were examined by one- and two-dimensional gel electrophoresis, and identified by peptide fingerprinting-mass spectrometry. A membrane-bound, externally oriented, quinone-linked formate dehydrogenase complex was expressed at high level specifically during syntrophic butyrate oxidation, comprising a selenocystein-linked catalytic subunit with a membrane-translocation pathway signal (TAT), a membrane-bound iron-sulfur subunit, and a membrane-bound cytochrome. Soluble hydrogenases were expressed at high levels specifically during growth with crotonate. The results were confirmed by native protein gel electrophoresis, by formate dehydrogenase and hydrogenase-activity staining, and by analysis of formate dehydrogenase and hydrogenase activities in intact cells and cell extracts. Furthermore, constitutive expression of a membrane-bound, internally oriented iron-sulfur oxidoreductase (DUF224) was confirmed, together with expression of soluble electron-transfer flavoproteins (EtfAB) and two previously identified butyryl-CoA dehydrogenases. Our findings allow to depict an electron flow scheme for syntrophic butyrate oxidation in S. wolfei. Electrons derived from butyryl-CoA are transferred through a membrane-bound EtfAB:quinone oxidoreductase (DUF224) to a menaquinone cycle and further via a b-type cytochrome to an externally oriented formate dehydrogenase. Hence, an ATP hydrolysis-driven proton-motive force across the cytoplasmatic membrane would provide the energy input for the electron potential shift necessary for formate formation.


Basolateral localisation of KCNQ1 potassium channels in MDCK cells: molecular identification of an N-terminal targeting motif.
KCNQ1 potassium channels are expressed in many epithelial tissues as well as in the heart. In epithelia KCNQ1 channels play an important role in salt and water transport and the channel has been reported to be located apically in some cell types and basolaterally in others. Here we show that KCNQ1 channels are located basolaterally when expressed in polarised MDCK cells. The basolateral localisation of KCNQ1 is not affected by co-expression of any of the five KCNE beta-subunits. We characterise two independent basolateral sorting signals present in the N-terminal tail of KCNQ1. Mutation of the tyrosine residue at position 51 resulted in a non-polarized steady-state distribution of the channel. The importance of tyrosine 51 in basolateral localisation was emphasized by the fact that a short peptide comprising this tyrosine was able to redirect the p75 neurotrophin receptor, an otherwise apically located protein, to the basolateral plasma membrane. Furthermore, a di-leucine-like motif at residues 38-40 (LEL) was found to affect the basolateral localisation of KCNQ1. Mutation of these two leucines resulted in a primarily intracellular localisation of the channel.


Role of lipids in the translocation of proteins across membranes.
The architecture of cells, with various membrane-bound compartments and with the protein synthesizing machinery confined to one location, dictates that many proteins have to be transported through one or more membranes during their biogenesis. A lot of progress has been made on the identification of protein translocation machineries and their sorting signals in various organelles and organisms. Biochemical characterization has revealed the functions of several individual protein components. Interestingly, lipid components were also found to be essential for the correct functioning of these translocases. This led to the idea that there is a very intimate relationship between the lipid and protein components that enables them to fulfil their intriguing task of transporting large biopolymers through a lipid bilayer without leaking their contents. In this review we focus on the Sec translocases in the endoplasmic reticulum and the bacterial inner membrane. We also highlight the interactions of lipids and proteins during the process of translocation and integrate this into a model that enables us to understand the role of membrane lipid composition in translocase function.


Inter-laboratory comparison study on measuring semi-volatile organic chemicals in standards and air samples.
Measurements of semi-volatile organic chemicals (SVOCs) were compared among 21 laboratories from 7 countries through the analysis of standards, a blind sample, an air extract, and an atmospheric dust sample. Measurement accuracy strongly depended on analytes, laboratories, and types of standards and samples. Intra-laboratory precision was generally good with relative standard deviations (RSDs) of triplicate injections <10% and with median differences of duplicate samples between 2.1 and 22%. Inter-laboratory variability, measured by RSDs of all measurements, was in the range of 2.8-58% in analyzing standards, and 6.9-190% in analyzing blind sample and air extract. Inter-laboratory precision was poorer when samples were subject to cleanup processes, or when SVOCs were quantified at low concentrations. In general, inter-laboratory differences up to a factor of 2 can be expected to analyze atmospheric SVOCs. When comparing air measurements from different laboratories, caution should be exercised if the data variability is less than the inter-laboratory differences.


Fungal asthma among Ugandan adult asthmatics.
Fungal sensitization is associated with poor asthma control. We aimed to determine the prevalence and factors associated with fungal asthma among Ugandan adults. Individuals aged ≥18 years with a new diagnosis of asthma in the last 12 months participating in the African Severe Asthma Program constituted the study population. Skin prick test results, clinical and demographic data were retrieved from the database, and serum Aspergillus fumigatus specific antibodies and total IgE were measured in stored blood. We enrolled 374 patients, median (IQR) age 34 (25-45) years, 286 (76.5%) females and 286 (76.5%) with severe asthma. Prevalence of Aspergillus fumigatus sensitization was 42.0% (95% CI: 37.1-47.0%), allergic bronchopulmonary aspergillosis (ABPA) 3.2% (1.8-5.5%), severe asthma with fungal sensitization (SAFS) 16% (12.7-20.1%) and allergic bronchopulmonary mycosis (ABPM) 2.9% (1.7-5.2%). Older age (55-64 years) (crude odds ratio (cOR) = 2.6), sensitization to at least one allergen (cOR = 9.38) and hypertension (cOR = 1.99) were significantly associated with Aspergillus sensitization, whereas tertiary education level (cOR = 0.29), severe depression (cOR = 0.15) and strong emotions (cOR = 0.47) were not. High occupational exposure to Aspergillus (cOR = 4.26) and contact with moulds (cOR = 14.28) were significantly associated with ABPA. Palpitations (cOR = 5.54), uncontrolled asthma (cOR = 3.54), eczema/dermatitis (cOR = 3.07), poor lung function (cOR = 2.11) and frequent exacerbations (cOR = 1.01) were significantly associated with SAFS. Eczema/dermatitis (cOR = 1.55) was significantly associated with ABPM, but cold weather trigger (cOR = 0.24) was not. Fungal asthma is a significant problem among Ugandans with asthma and should be particularly considered in individuals who remain uncontrolled despite optimal standard of care for asthma, as it is responsive to available and affordable oral antifungal therapy. This study showed that fungal asthma is a significant problem among Ugandans with asthma with a high prevalence. Fungal asthma should be considered in patients with uncontrolled asthma despite receiving optimal standard of care. This is the first modern attempt to define these endotypes of asthma in Africa.


Factors predicting cardiovascular events in chronic kidney disease patients. Role of subclinical atheromatosis extent assessed by vascular ultrasound.
Patients with chronic kidney disease (CKD) have an increased incidence of cardiovascular events (CVE). The contribution of subclinical atheromatosis extent, including femoral arteries, to CVE in CKD patients has not been investigated. In this paper, we examine the prognostic value of subclinical atheromatosis extent, assessed as the number of arterial territories with plaque, in predicting the incidence of major and minor CVE. The NEFRONA is a multicenter, prospective cohorts study that recruited 2445 CKD subjects and 559 controls, free from previous cardiovascular disease, in 81 medical centers across Spain. The presence of atheroma plaque was assessed by arterial ultrasound in ten arterial territories (carotid and femoral). The predictive power of the presence or absence of atheroma plaque in any territory was compared with the quantification of atheroma extent as the number of territories with plaque. During the median follow up of 48 months, 216 CVE were reported. Factors predicting the incidence of CVE in the whole cohort were being male, CKD patient, lower levels of 25(OH) vitamin D, higher levels of cholesterol and the extent of subclinical atheromatosis, yielding a higher concordance (C) index than the presence or absence of plaque. In stratified analysis including specific factors of CKD patients not on dialysis, the variables predicting CVE were the same as in the whole cohort, plus higher levels of potassium. Again, the inclusion of the information about atheromatosis as number of territories with plaque, presented a higher C index than the presence or absence of plaque. In the dialysis population, significant variables were older age, diabetes, dialysis vintage and higher levels of cholesterol and phosphate. In this case the higher C index was obtained with the information about plaque presence. Subclinical atheromatosis extent, including femoral arteries, influences CVE in CKD and its detection could improve the prediction of cardiovascular events.


Ten-Month-Old Infants' Reaching Choices for "more": The Relationship between Inter-Stimulus Distance and Number.
Animals and human infants discriminate numerosities in visual sets. Experiments on visual numerical judgments generally contrast sets in which number varies (e.g., the discrimination between 2 and 3). What is less investigated, however, is set density, or rather, the inter-stimulus distance between the entities being enumerated in a set. In this study, we investigated the role of set density in visual sets by 10-month-old infants. In Experiment 1, infants were offered a choice between two sets each containing four items of the exact same size varying in the distance in between the items (ratio 1:4). Infants selected the set in which the items are close together (higher density). Experiment 2 addressed the possibility that this choice was driven by a strategy to "select all in one go" by reducing the size and distance of items. Ten-month-olds selected the sets with higher density (less inter-stimulus distance) in both experiments. These results, although bearing replication because of their originality, seem consistent with principles in Optimal Foraging in animals. They provide evidence that a comparable rudimentary capacity for density assessment (of food items) exists in infants, and may work in concert with their numerical representations.


Dynamic remodeling of subcellular chemical gradients using a multi-directional flow device.
Elucidation of the mechanisms by which external chemical cues regulate polarized cellular behaviors requires tools that can rapidly recast chemical landscapes with subcellular resolution. Here, we describe an approach for creating steep microscopic gradients of cellular effectors at any desired position in culture that can be reoriented rapidly to evaluate dynamic responses. In this approach, micrometre pores are ablated in a membrane that supports cell adherence, allowing dosing reagent from an underlying reservoir to enter the cell-culture flow chamber as sharp streams that are directed at subcellular targets by using a system of paired sources and drains to specify flow direction. This tool substantially extends capabilities for chemical interaction with cultured cells, enabling investigations of chemotaxis via precise placement and reorientation of peptide gradients formed at the boundaries of dosing streams. These studies demonstrate that neutrophil precursor cells can repolarize and redirect their migration paths using morphological responses that depend on the subcellular localization of chemoattractant gradients.


Genetics of Alzheimer's Disease in the African American Population.
Black/African American (AA) individuals have a higher risk of Alzheimer's disease (AD) than White non-Hispanic persons of European ancestry (EUR) for reasons that may include economic disparities, cardiovascular health, quality of education, and biases in the methods used to diagnose AD. AD is also heritable, and some of the differences in risk may be due to genetics. Many AD-associated variants have been identified by candidate gene studies, genome-wide association studies (GWAS), and genome-sequencing studies. However, most of these studies have been performed using EUR cohorts. In this paper, we review the genetics of AD and AD-related traits in AA individuals. Importantly, studies of genetic risk factors in AA cohorts can elucidate the molecular mechanisms underlying AD risk in AA and other populations. In fact, such studies are essential to enable reliable precision medicine approaches in persons with considerable African ancestry. Furthermore, genetic studies of AA cohorts allow exploration of the ways the impact of genes can vary by ancestry, culture, and economic and environmental disparities. They have yielded important gains in our knowledge of AD genetics, and increasing AA individual representation within genetic studies should remain a priority for inclusive genetic study design.


Induced defensive response of myrtle oak to foliar insect herbivory in ambient and elevated CO2.
The rising level of atmospheric CO2 has stimulated several recent studies attempting to predict the effects of increased CO2 on ecological communities. However, most of these studies have been conducted in the benign conditions of the laboratory and in the absence of herbivores. In the current study, we utilized large octagonal chambers, which enclosed portions of an intact scrub-oak community to investigate the interactive effects of CO2 and insect herbivory on myrtle oak, Quercus myrtifolia. Specifically, we assessed the effects of ambient and elevated CO2 (2x current concentrations) on percent foliar nitrogen, C:N ratio, total relative foliar tannin content, and the presence of leaf damage caused by leaf mining and leaf chewing insects that feed on myrtle oak. Total foliar N declined and C:N ratios increased significantly in oaks in elevated CO2 chambers. The percentages of leaves damaged by either leafminers or leaf chewers tended to be lower in elevated compared to ambient chambers, but they co-occurred on leaves less than expected, regardless of CO2 treatment. Leaves that had been either mined or chewed exhibited a similar wounding or defensive response; they had an average of 25 and 21% higher protein binding ability, which is correlated with tannin concentration, compared to nondamaged control leaves, respectively. While the protein-binding ability (expressed as total percent tannin) of leaves from elevated CO2 was slightly higher than from leaves grown in ambient chambers, this difference was not significant.


Identification of a domain in the V0 subunit d that is critical for coupling of the yeast vacuolar proton-translocating ATPase.
Vacuolar proton-translocating ATPase pumps consist of two domains, V(1) and V(o). Subunit d is a component of V(o) located in a central stalk that rotates during catalysis. By generating mutations, we showed that subunit d couples ATP hydrolysis and proton transport. The mutation F94A strongly uncoupled the enzyme, preventing proton transport but not ATPase activity. C-terminal mutations changed coupling as well; ATPase activity was decreased by 59-72%, whereas proton transport was not measurable (E328A) or was moderately reduced (E317A and C329A). Except for W325A, which had low levels of V(1)V(o), mutations allowed wild-type assembly regardless of the fact that subunits E and d were reduced at the membrane. N- and C-terminal deletions of various lengths were inhibitory and gradually destabilized subunit d, limiting V(1)V(o) formation. Both N and C terminus were required for V(o) assembly. The N-terminal truncation 2-19Delta prevented V(1)V(o) formation, although subunit d was available. The C terminus was required for retention of subunits E and d at the membrane. In addition, the C terminus of its bacterial homolog (subunit C from T. thermophilus) stabilized the yeast subunit d mutant 310-345Delta and allowed assembly of the rotor structure with subunits A and B. Structural features conserved between bacterial and eukaryotic subunit d and the significance of domain 3 for vacuolar proton-translocating ATPase function are discussed.


Active ras triggers death in glioblastoma cells through hyperstimulation of macropinocytosis.
Expression of activated Ras in glioblastoma cells induces accumulation of large phase-lucent cytoplasmic vacuoles, followed by cell death. This was previously described as autophagic cell death. However, unlike autophagosomes, the Ras-induced vacuoles are not bounded by a double membrane and do not sequester organelles or cytoplasm. Moreover, they are not acidic and do not contain the autophagosomal membrane protein LC3-II. Here we show that the vacuoles are enlarged macropinosomes. They rapidly incorporate extracellular fluid-phase tracers but do not sequester transferrin or the endosomal protein EEA1. Ultimately, the cells expressing activated Ras detach from the substratum and rupture, coincident with the displacement of cytoplasm with huge macropinosome-derived vacuoles. These changes are accompanied by caspase activation, but the broad-spectrum caspase inhibitor carbobenzoxy-Val-Ala-Asp-fluoromethylketone does not prevent cell death. Moreover, the majority of degenerating cells do not exhibit chromatin condensation typical of apoptosis. These observations provide evidence for a necrosis-like form of cell death initiated by dysregulation of macropinocytosis, which we have dubbed "methuosis." An activated form of the Rac1 GTPase induces a similar form of cell death, suggesting that Ras acts through Rac-dependent signaling pathways to hyperstimulate macropinocytosis in glioblastoma. Further study of these signaling pathways may lead to the identification of other chemical and physiologic triggers for this unusual form of cell death.


Differences between younger and older individuals in their use of care and rehabilitation but not in self-perceived global recovery 1year after stroke.
Although priorities in Swedish stroke care should be based on the ethical principles of equal care and greatest support to those in greatest need, being of working age (younger) or retired (older) might influence expectations on recovery and the provision of care and rehabilitation. Information regarding the use of care and rehabilitation during the 1st year after stroke was retrieved from the Stockholm County Council database and the medical data was taken from the medical records. The Barthel Index was used for self-ratings of dependence pre-stroke, and the Stroke Impact Scale was used to assess self-perceived disability and a global rating of recovery at 12 months. One hundred and ninety-two individuals were included. Results showed that the younger group received more care and rehabilitation than the older group. In the older group, comorbid conditions and pre-stroke dependence, estimated in accordance with the Barthel Index, were more common. The older group reported larger impact on self-perceived disability regarding strength, mobility, self-care and domestic life, while self-perceived global recovery did not differ between the groups. Younger individuals received more care and rehabilitation, which indicates structural inequality in the provision of health care resources. However, as no difference in self-perceived global recovery was found between the groups; the disparity in the provision of health care may also be a consequence of greatest support being given to those in greatest need. By demonstrating the necessity of involving self-rated assessments of stroke impact at different points in time and in different age groups, the results of this study add to previous knowledge of age-related differences in the provision of health care and stroke outcome. Furthermore, this suggests a way to approach evaluation of equality in the provision of health care.


Botulinum Neurotoxins: Biology, Pharmacology, and Toxicology.
The study of botulinum neurotoxins (BoNT) is rapidly progressing in many aspects. Novel BoNTs are being discovered owing to next generation sequencing, but their biologic and pharmacological properties remain largely unknown. The molecular structure of the large protein complexes that the toxin forms with accessory proteins, which are included in some BoNT type A1 and B1 pharmacological preparations, have been determined. By far the largest effort has been dedicated to the testing and validation of BoNTs as therapeutic agents in an ever increasing number of applications, including pain therapy. BoNT type A1 has been also exploited in a variety of cosmetic treatments, alone or in combination with other agents, and this specific market has reached the size of the one dedicated to the treatment of medical syndromes. The pharmacological properties and mode of action of BoNTs have shed light on general principles of neuronal transport and protein-protein interactions and are stimulating basic science studies. Moreover, the wide array of BoNTs discovered and to be discovered and the production of recombinant BoNTs endowed with specific properties suggest novel uses in therapeutics with increasing disease/symptom specifity. These recent developments are reviewed here to provide an updated picture of the biologic mechanism of action of BoNTs, of their increasing use in pharmacology and in cosmetics, and of their toxicology.


Hepatoprotective effects of aqueous extract from Lingzhi or Reishi medicinal mushroom Ganoderma lucidum (higher basidiomycetes) on α-amanitin-induced liver injury in mice.
The Lingzhi or Reishi mushroom Ganoderma lucidum is a well-known traditional medicinal mushroom that has been shown to have obvious hepatoprotective effects. The aim of this study was to evaluate the hepatoprotective effects of G. lucidum aqueous extracts (GLEs) on liver injury induced by α-amanitin (α-AMA) in mice and to analyze the possible hepatoprotective mechanisms related to radical scavenging activity. Mice were treated with α-AMA prepared from Amanita exitialis and then administrated with GLE after the α-AMA injection. The hepatoprotective activity of the GLE was compared with the reference drug silibinin (SIL). α-AMA induced a significant elevation in serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities and provoked a significant reduction of superoxide dismutase (SOD) and catalase (CAT) activities and a significant increment of malondialdehyde (MDA) content in liver homogenate. Treatment with GLE or SIL significantly decreased serum ALT and AST levels, significantly increased SOD and CAT activities, and decreased MDA content in liver compared with the α-AMA control group. The histopathological examination of liver sections was consistent with that of biochemical parameters. The results demonstrated that GLE induces hepatoprotective effects on acute liver injury induced by α-AMA; these protective effects may be related in part to the antioxidant properties of GLE.


Prenatal and postnatal psychological symptoms of parents and family functioning: the impact on child emotional and behavioural problems.
Although relations of various parental psychological problems and family functioning with child development are well documented, it remains unclear whether specific prenatal or specific postnatal risk factors are independently associated with child emotional and behavioural problems, or whether observed associations can be explained by general parental psychopathology. Using a stepwise approach, we examined the effects of prenatal and postnatal parental depressive symptoms, prenatal and postnatal hostility of the parents, as well as prenatal family functioning on the risk of child emotional and behavioural problems. This study was embedded in Generation R: a population-based cohort from foetal life onwards. Mothers and fathers of 2,698 children provided information about depressive symptoms, symptoms of hostility and family functioning during pregnancy and 3 years after birth. Mother and father each reported on child behaviour when the child was 3 years old. Parental depressive symptoms increased the risk of child emotional and behavioural problems, but this increase was explained by postnatal parental hostile behaviour. Postnatal symptoms of hostility of mothers (OR = 1.34, p value <0.001) and postnatal symptoms of hostility of fathers (OR = 1.30, p value <0.001) each contributed independently to the risk of child emotional and behavioural problems. Postnatal parental hostility is associated with an increased risk of child emotional and behavioural problems, independent of parental depressive symptoms. These findings suggest that prevention and intervention strategies should focus on psychological symptoms of both mothers and fathers, in particular on hostile behaviour, in families with young children.


Replacement of the failing heart.
Heart transplantation is an effective means of treating patients with severe congestive heart failure. Following heart transplantation, the 1-year survival rate is now greater than 80%, and the 5-year survival rate is more than 60% at major medical centers. More than 1,200 heart transplants were performed in more than nine countries worldwide in 1985. The failure of medicare to pay for this procedure is no longer defensible on medical grounds. The argument in favor of medicare funding for heart transplantation is at least as compelling as that for kidney dialysis, the treatment of cancer, or AIDS. The limited availability of donor organs (at most, 1300-2000/year) is likely to place a finite constraint on the number of heart transplants that can and will be performed. Although combined heart-lung transplantation is feasible therapy for certain patients with severe pulmonary hypertension, the availability of suitable donors poses an even greater restriction on this procedure. Totally implantable ventricular assist devices are on the horizon. These devices have the potential for helping 17,000 to 35,000 patients annually at an estimated cost to society of $2.5 to $5 billion per annum. The development and use of such extremely expensive technology poses major socioeconomic and ethical questions for society.


What do eye movements tell us about the visual perception of individuals with congenital prosopagnosia?
The lack of inversion effect for face recognition in congenital prosopagnosia (CP) is consistent with the hypothesis of a failure in holistic processing. However, although CPs' abnormal gaze behavior for upright faces has already been demonstrated, neither their scanning strategy for inverted faces, nor the possibility that their abnormal gaze behavior with upright faces is because of reasons other than the holistic deficit have been investigated yet. We recorded the eye movements of a congenital prosopagnosic and a control group during the encoding of unknown faces, objects, and flowers. Two types of stimuli (faces and objects) were presented upright and inverted. CPs explored upright and inverted faces in the same way (i.e., similar number of fixations of the same duration and similarly distributed), whereas controls increased the number of fixations and their duration during the presentation of inverted faces. By contrast, the 2 groups showed a similar inversion effect during the encoding of objects. Finally, CPs showed anomalous exploration of within-class objects (i.e., flowers) and impairment in subordinate-level object discrimination. Our results demonstrate that: (a) CPs use the same part-based strategy in encoding both upright and inverted faces, suggesting a possible interpretation of the lack of inversion effect in this population; (b) CPs' lack of inversion effect is face-specific and does not affect objects; (c) however, CPs' deficit seems not to be limited to faces, and to extend to individual-item recognition within a class. (PsycINFO Database Record


Reappraisal of intergender differences in the urethral striated sphincter explains why a completely circular arrangement is difficult in females: a histological study using human fetuses.
To investigate why the development of a completely circular striated sphincter is so rare, we examined histological sections of 11 female and 11 male mid-term human fetuses. In male fetuses, the striated muscle initially extended in the frontal, rather than in the horizontal plane. However, a knee-like portion was absent in the female fetal urethra because, on the inferior side of the vaginal end, a wide groove for the future vestibule opened inferiorly. Accordingly, it was difficult for the developing striated muscle to surround the groove, even though there was not a great difference in width or thickness between the female vestibule and the male urethra. The development of a completely circular striated sphincter seems to be impossible in females because of interruption of the frontal plane by the groove-like vestibule. However, we cannot rule out the possibility that before descent of the vagina, the urethral striated muscle extends posteriorly.


Medico-legal claims in colorectal surgery: Analysis of 231 files.
The patient undergoing surgery is at risk of complications, some of which can be serious; these can expose the surgeon or institution to claims for compensation of damages. The aim of this study is to analyze the surgical management of these situations in order to draw lessons, to improve the management of patients, and to prevent both complications and resulting claims. Two visceral surgeons analyzed two hundred and thirty-one claims files. The patient's characteristics and data, the intervention, the source event of the claim, re-interventions, transfers, deaths and their causes were identified. Any error, whether it was identified by the expert or by the arbitration panel or court and vice versa was analyzed, including any amicable out-of-court agreement proposed by the insurance, even, if in certain cases, no real fault had been found. The mean age of the colon surgery patients was 62 years. The pathologies for which surgery was performed fell into three equal parts: colon cancer, sigmoid diverticulitis, and other pathologies. The event leading to the claim occurred during hospitalization in 69.2% of cases. The most common events prompting claims were anastomotic leak (34.1%) and injury to neighboring organs (16.4%). In 36.7% of cases, patients required transfer to another facility and 31.1% died. At least one fault or error was found in 46.8% of cases. Anastomotic leak has always been considered an inherent risk of colonic surgery. The main fault alleged was delay in management in nearly two out of three cases, including delays in communication, in physical and laboratory examination, medical treatment, re-operation, and transfer. Specific information was provided to the patient before surgery and an effective checklist to prevent complaints and complications. In the post-operative period, an active approach to management initiated without delay by the entire team when faced with any unexpected event favored a quick recovery and could avoid the complaints; exams, sometimes lacking, remain complementary as their names indicate.


Increased Early Systemic Inflammation in ICU-Acquired Weakness; A Prospective Observational Cohort Study.
To investigate whether patients who develop ICU-acquired weakness have a different pattern of systemic inflammatory markers compared with critically ill patients who do not develop ICU-acquired weakness. Prospective observational cohort study. Mixed medical-surgical ICU of a tertiary care hospital in the Netherlands. Newly admitted critically ill patients, greater than or equal to 48 hours on mechanical ventilation with a nonneurologic ICU admission diagnosis, were included. A panel of systemic inflammatory markers and soluble vascular adhesion molecules were measured in plasma samples of day 0, 2, and 4 after ICU admission. ICU-acquired weakness was diagnosed by manual muscle strength testing as soon as patients were awake and attentive. Ninety-nine of 204 included patients developed ICU-acquired weakness. Principal component regression analysis, adjusted for confounders, showed that principal component 1, mainly loaded with interleukin-6, interleukin-8, interleukin-10, and fractalkine, was significantly higher in patients who developed ICU-acquired weakness (odds ratio, 1.35 [95% CI, 1.18-1.55]). Partial least squares-discriminant analysis also showed that these markers were the most important discriminative markers. Mixed-effects models of these markers showed that ICU-acquired weakness was associated with an independent 1.5- to two-fold increase in these markers. Systemic inflammation is increased in patients who develop ICU-acquired weakness compared with patients who do not develop ICU-acquired weakness in the first 4 days after ICU admission. This finding is consistent when adjusted for confounders, like disease severity. A group consisting of interleukin-6, interleukin-8, interleukin-10, and fractalkine was identified to be the most important.


Validity and Reliability of the Turkish Version of Mandibular Function Impairment Questionnaire.
To assess the validity and reliability of the Turkish version of the Mandibular Function Impairment Questionnaire (MFIQ-TR). Included in this study were 248 patients diagnosed with temporomandibular disorder (TMD) according to Diagnostic Criteria for TMD Axis I protocol. Construct-related validity was evaluated through internal and external construct validity; convergent and divergent validities were evaluated by the average variance extracted (AVE), composite reliability (CR), and bivariate correlations between factors. Reliability was assessed by internal consistency and test-retest reliability. All items of the MFIQ-TR had content validity and factor loadings above 0.5. The model statistics indicated a good fit. The convergent validity was very good (AVE > 0.5 and CR > 0.7), and the discriminant validity was satisfied. Internal consistency and test-retest reliability of the MFIQ-TR were excellent. The MFIQ-TR is a valid and reliable instrument for evaluating the impairment of mandibular function in Turkish patients with TMD.


Impact of A1C screening criterion on the diagnosis of pre-diabetes among U.S. adults.
New clinical practice recommendations include A1C as an alternative to fasting glucose as a diagnostic test for identifying pre-diabetes. The impact of these new recommendations on the diagnosis of pre-diabetes is unknown. Data from the National Health and Nutrition Examination Survey 1999-2006 (n = 7,029) were analyzed to determine the percentage and number of U.S. adults without diabetes classified as having pre-diabetes by the elevated A1C (5.7-6.4%) and by the impaired fasting glucose (IFG) (fasting glucose 100-125 mg/dl) criterion separately. Test characteristics (sensitivity, specificity, and positive and negative predictive values) using IFG as the reference standard were calculated. The prevalence of pre-diabetes among U.S. adults was 12.6% by the A1C criterion and 28.2% by the fasting glucose criterion. Only 7.7% of U.S. adults, reflecting 61 and 27% of those with pre-diabetes by A1C and fasting glucose, respectively, had pre-diabetes according to both definitions. A1C used alone would reclassify 37.6 million Americans with IFG to not having pre-diabetes and 8.9 million without IFG to having pre-diabetes (46.5 million reclassified). Using IFG as the reference standard, pre-diabetes by the A1C criterion has 27% sensitivity, 93% specificity, 61% positive predictive value, and 77% negative predictive value. Using A1C as the pre-diabetes criterion would reclassify the pre-diabetes diagnosis of nearly 50 million Americans. It is imperative that clinicians and health systems understand the differences and similarities in using A1C or IFG in diagnosis of pre-diabetes.


Quality-of-Life in Achilles Tendinopathy: An Exploratory Study.
This research aimed to explore the quality-of-life and experiences of people with Achilles tendinopathy. This mixed-methods research used the 8-dimension Assessment of Quality-of-Life (AQoL-8D), focus groups and grounded theory analysis. AQoL-8D scores were compared with population normative scores. In focus groups, participants discussed their experiences with Achilles tendinopathy. An online survey was completed, followed by focus groups and interviews held at the University of Canberra. Adults with Achilles tendon pain were eligible to participate in the online survey, which was distributed through email and social media. Complete survey responses were obtained from 92 individuals, and 11 individuals participated in focus groups and interviews. AQoL-8D scores were significantly lower in those with Achilles tendinopathy (79 ± 11 vs 81 ± 13). AQoL-8Ds of mental health, pain, senses, and the physical "super dimension" were also significantly lower. The difference exceeded the AQoL-8D minimum clinically important difference of 6% only for the pain dimension. Themes identified included adapting lifestyles, living with the condition, changes in mental and social well-being, conflict with identity, frustration, and individual experiences. Achilles tendinopathy is associated with a lower quality-of-life score, but on average, the difference does not exceed the minimum clinically important difference. In focus groups, some individuals described profound impacts on their life. This discrepancy likely reflects the variability of the impact across individuals. For some people, the effect is minimal, yet for those who tie their identity and social activities to fitness and physical activity, the effect can be profound.


Comparison of staff and family perceptions of causes of noise pollution in the Pediatric Intensive Care Unit and suggested intervention strategies.
Noise and excessive, unwanted sound in the Pediatric Intensive Care Unit (PICU) is common and has a major impact on patients' sleep and recovery. Previous research has focused mostly on absolute noise levels or included only staff as respondents to acknowledge the causes of noise and to plan for its reduction. Thus far, the suggested interventions have not ameliorated noise, and it continues to serve as a barrier to recovery. In addition to surveying PICU providers through internet-based software, patients' families were evaluated through in-person interviews utilizing a pretested instrument over 3 months. Families of patients admitted for more than 24 h were considered eligible for evaluation. Participants were asked to rank causes of noise from 1 to 8, with eight being highest, and identified potential interventions as effective or ineffective. In total, 50 families from 251 admissions and 65 staff completed the survey. Medical alarms were rated highest (mean ± standard deviation [SD], 4.9 ± 2.1 [2.8-7.0]), followed by noise from medical equipment (mean ± SD, 4.7 ± 2.1 [2.5-6.8]). This response was consistent among PICU providers and families. Suggested interventions to reduce noise included keeping a patient's room door closed, considered effective by 93% of respondents (98% of staff; 88% of families), and designated quiet times, considered effective by 82% (80% of staff; 84% of families). Keeping the patient's door closed was the most effective strategy among survey respondents. Most families and staff considered medical alarms an important contributor to noise level. Because decreasing the volume of alarms such that it cannot be heard is inappropriate, alternative strategies to alert staff of changes in vital signs should be explored.


Objective and subjective analysis of left ventricular angiograms.
In order to determine the reproducibility of analyses of left ventriculograms, 35 cineangiograms were evaluated by four observers, two using standard quantitative techniques to determine ventricular volumes and a newly devised quantitative system to evaluate wall motion and two others using only visual inspection of the angiograms. Objective analysis repeated by the same observer correlated well for end-diastolic and end-systolic volumes and ejection fraction (r = .98, .99, respectively) and only one of 105 (1%) wall segments were identified differently. Variability in assessments increased when comparisons were made with a second objective observer. Correlation coefficients for the three volumetric parameters were .93, .98, and .95 and there was disagreement in the assessment of 8% of wall segments. Wide variability was present between an objective and two subjective observers in analyses of end-diastolic volumes (r = .63, .64). Regional wall motion was assessed differently in 19% and 27% of segments, respectively. Though the correlation of objectively and subjectively determined ejection fractions was much better than the correlation for volume (r = .92, .84), it was not as good as the correlation between two objective observers. Occasional errors of clinical significance occurred. We conclude that subjective analysis has a significant error rate and that reproducibility and accuracy of analysis of left ventriculograms require objective analysis.


A Population Pharmacokinetic-Pharmacodynamic Model of Pegfilgrastim.
Neutropenia and febrile neutropenia (FN) are serious side effects of cytotoxic chemotherapy which may be alleviated with the administration of recombinant granulocyte colony-stimulating factor (GCSF) derivatives, such as pegfilgrastim (PG) which increases absolute neutrophil count (ANC). In this work, a population pharmacokinetic-pharmacodynamic (PKPD) model was developed based on data obtained from healthy volunteers receiving multiple administrations of PG. The developed model was a bidirectional PKPD model, where PG stimulated the proliferation, maturation, and margination of neutrophils and where circulating neutrophils in turn increased the elimination of PG. Simulations from the developed model show disproportionate changes in response with changes in dose. A dose increase of 10% from the 6 mg therapeutic dose taken as a reference leads to area under the curve (AUC) increases of ~50 and ~5% for PK and PD, respectively. A full random effects covariate model showed that little of the parameter variability could be explained by sex, age, body size, and race. As a consequence, little of the secondary parameter variability (Cmax and AUC of PG and ANC) could be explained by these covariates.


Should intentional endovascular stent-graft coverage of the left subclavian artery be preceded by prophylactic revascularisation?
Thoracic endovascular aortic repair (TEVAR) has emerged as a promising therapeutic alternative to conventional open aortic replacement but it requires suitable proximal and distal landing zones for stent-graft anchoring. Many aortic pathologies affect in the immediate proximity of the left subclavian artery (LSA) limiting the proximal landing zone site without proximal vessel coverage. In patients in whom the distance between the LSA and aortic lesion is too short, extension of the landing zone can be obtained by covering the LSA's origin with the endovascular stent graft (ESG). This manoeuvre has the potential for immediate and delayed neurological and vascular symptoms. Some authors, therefore, propose prophylactic revascularisation of the LSA by transposition or bypass, while others suggest prophylactic revascularisation only under certain conditions, and still others see no requirement for prophylactic revascularisation in anticipation of LSA ostium coverage. In this review about LSA revascularisation in TEVAR patients with coverage of the LSA, we searched the electronic databases MEDLINE and EMBASE historically until the end date of May 2010 with the search terms left subclavian artery, covering, endovascular, revascularisation and thoracic aorta. We have gathered the most complete scientific evidence available used to support the various concepts to deal with this issue. After a review of the current available literature, 23 relevant articles were found, where we have identified and analysed three basic treatment concepts for LSA revascularisation in TEVAR patients (prophylactic, conditional prophylactic and no prophylactic LSA revascularisation). The available evidence supports prophylactic revascularisation of the LSA before ESG LSA coverage when preoperative imaging reveals abnormal supra-aortic vascular anatomy or pathology. We further conclude that elective patients undergoing planned coverage of the LSA during TEVAR should receive prophylactic LSA transposition or LSA-to-left-common-carotid-artery (LCCA) bypass surgery to prevent severe neurological complications, such as paraplegia or brain stem infarction.


Selection and transformation of non-beta-lactamase-mediated insusceptibility to beta-lactams in Haemophilus influenzae: lack of cross-resistance between carbapenems and other agents.
Non-beta-lactamase-mediated insusceptibility to beta-lactams was studied in Haemophilus influenzae. Mutants resistant to ampicillin were selected in a susceptible isolate, while phenotypically similar resistance was transformed from four wild-type isolates into a susceptible recipient. Ampicillin-selected mutants generally had reduced susceptibility to penicillins, cephalosporins and aztreonam. Reduced susceptibility to meropenem was less common, and susceptibility to imipenem was unaffected. MICs of three cephalosporins and aztreonam were much higher for the ampicillin-resistant transformants than for the recipient strain, and were generally equal to, or within one doubling dilution of, the concentrations that inhibited the donor parents. In contrast, reduced susceptibility to imipenem observed with one parent (MIC of 4 mg/l) was not transferred to the recipient (MIC of 0.5 mg/l). Meropenem inhibited 22 of 24 transformants tested at less than or equal to 0.12 mg/l, compared with MICs of greater than or equal to 0.5 mg/l for the donor parents and 0.03 mg/l for the recipient. These results indicated that the mechanisms that confer non-beta-lactamase-mediated insusceptibility to penicillins, cephalosporins and aztreonam in H. influenzae have little or no effect on susceptibility to carbapenems.


[Analysis of rat enterocyte ultrastructure during glucose absorption].
Electronmicroscopic study of rat enterocytes under glucose load (10-40 mM) has shown some changes of their structure: aggregations of intramembrane particles of the apical membrane in the microvilli region, the dilitation of lateral intercellular spaces below tight junction, the condensation of actin near tight and intermediate junctions. The presence of these changes and almost absolute absence of destructions in tight junctions organization indicate that the main pathway of the isotonic fluid containing glucose across leaky epithelium of rat small intestine is a transcellular one.


Characterization of erythroferrone structural domains relevant to its iron-regulatory function.
Iron delivery to the plasma is closely coupled to erythropoiesis, the production of red blood cells, as this process consumes most of the circulating plasma iron. In response to hemorrhage and other erythropoietic stresses, increased erythropoietin stimulates the production of the hormone erythroferrone (ERFE) by erythrocyte precursors (erythroblasts) developing in erythropoietic tissues. ERFE acts on the liver to inhibit bone morphogenetic protein (BMP) signaling and thereby decrease hepcidin production. Decreased circulating hepcidin concentrations then allow the release of iron from stores and increase iron absorption from the diet. Guided by evolutionary analysis and Alphafold2 protein complex modeling, we used targeted ERFE mutations, deletions, and synthetic ERFE segments together with cell-based bioassays and surface plasmon resonance to probe the structural features required for bioactivity and BMP binding. We define the ERFE active domain and multiple structural features that act together to entrap BMP ligands. In particular, the hydrophobic helical segment 81 to 86 and specifically the highly conserved tryptophan W82 in the N-terminal region are essential for ERFE bioactivity and Alphafold2 modeling places W82 between two tryptophans in its ligands BMP2, BMP6, and the BMP2/6 heterodimer, an interaction similar to those that bind BMPs to their cognate receptors. Finally, we identify the cationic region 96-107 and the globular TNFα-like domain 186-354 as structural determinants of ERFE multimerization that increase the avidity of ERFE for BMP ligands. Collectively, our results provide further insight into the ERFE-mediated inhibition of BMP signaling in response to erythropoietic stress.


Role of membrane fusion in CO2 stimulation of proton secretion by turtle bladder.
The changes in cell structure produced during stimulation of proton secretion by CO2 in turtle bladder were examined using ultrastructural morphometric methods. One hour after CO2 addition, the area of the luminal membrane of the carbonic anhydrase-containing (CA) cell population was increased 2.5-fold and the volume percent of electron-lucent cytoplasmic vesicles in these CA cells was decreased by 61%. No changes were observed in the epithelial granular cells. These results suggest that during CO2 stimulation the vesicles fuse with the luminal membrane. CO2 stimulation of proton secretion is inhibited by the cytoskeleton-disrupting drugs colchicine and cytochalasin B and by 99% deuterium oxide as the Ringer solvent. Deuterium oxide also inhibits the decrease in cytoplasmic vesicles. Thus stimulation of proton secretion by turtle bladder CA cells depends to a large extent on vesicle fusion and the resultant increase in luminal surface area.


Differential expression of allograft inflammatory factor 1 and of glutathione peroxidase during auto- and allograft response in marine sponges.
Very recently, Porifera (sponges) have been proven to be suitable model systems to study auto- and allograft recognition at the molecular level. Several potential immune molecules have been isolated from the marine sponges Suberites domuncula and Geodia cydonium, among them those which comprise Ig-like domains in their extracellular part. Here we report on the isolation of two cDNAs from S. domuncula that code for molecules involved in mammals in cytokine-mediated graft response; a putative allograft inflammatory factor 1 (AIF-1) and a non-selenium glutathione peroxidase (GPX). Both polypeptides share high similarity with the corresponding mammalian proteins. The expression of the two genes during auto- and allograft recognition in S. domuncula and G. cydonium was determined. It is shown that the expression of the AIF-1-related gene is upregulated only in allografts, while the GPX-related gene is expressed in the fusion zones formed between auto- as well as allografts. Taken together, these findings suggest that besides cell-mediated defense reactions a cytokine-dependent immune response is also elicited during graft recognition in sponges.