[Culture-confirmed whooping cough in a tertiary center over a twelve-year period].
To study the characteristics of patients diagnosed with whooping cough at a tertiary center in Barcelona, Spain. We performed a retrospective study of patients aged less than 18 years treated for pertussis-like cough or clinically-suspected whooping cough over a 12-year period (1989-2000). Only patients with isolated Bordetella spp. were included. The variables of age, sex, vaccination status, hospitalization, clinical manifestations, severity, and lethality were analyzed. One hundred sixty-one patients with positive Bordetella spp. culture were identified. Of these, complete information was available in 149 (79 boys and 70 girls) with a median age of 3 months (range: 13 days-17 years); 77.2 % were aged 6 months or less. All the isolated strains corresponded to B. pertussis except three that corresponded to B. parapertussis. Three epidemic cycles (in 1989, 1992 and 2000) were observed during the study period. A total of 72.5 % of cases occurred between May and September. Bordetella spp. was associated with other bacteria in 28.2 % of the patients, viruses in 13.4 % and a bacterium and a virus in 4.7 %. One hundred twenty-one patients required hospitalization, of which 14.9 % were admitted to the intensive care unit. Age was the only factor associated with risk for hospitalization, which was more frequent in younger infants (p < 0.0001). Paroxysmal cough with cyanosis was present in 53.4 % of the patients, leucocytosis with lymphocytosis occurred in 67.5 % and apneas were present in 21.5 %. Chest X-ray revealed atelectasis in 34.1 %. The mean length of hospital stay was 11 days (range: 1-70 days). Three boys aged less than 3 months with malignant pertussis syndrome died (lethality: 2 %). More than half the patients (59.7 %) were not vaccinated (55.4 % for being under the age of 3 months) and only 16 % had received three or more vaccination doses. Whooping cough continues to be a severe disease in infants, with a high admission rate during the first 6 months of life. New preventive strategies are required to protect infants who have not yet developed full immunity to this infection.


Structure and function of factor XI.
Factor XI (FXI) is the zymogen of an enzyme (FXIa) that contributes to hemostasis by activating factor IX. Although bleeding associated with FXI deficiency is relatively mild, there has been resurgence of interest in FXI because of studies indicating it makes contributions to thrombosis and other processes associated with dysregulated coagulation. FXI is an unusual dimeric protease, with structural features that distinguish it from vitamin K-dependent coagulation proteases. The recent availability of crystal structures for zymogen FXI and the FXIa catalytic domain have enhanced our understanding of structure-function relationships for this molecule. FXI contains 4 "apple domains" that form a disk structure with extensive interfaces at the base of the catalytic domain. The characterization of the apple disk structure, and its relationship to the catalytic domain, have provided new insight into the mechanism of FXI activation, the interaction of FXIa with the substrate factor IX, and the binding of FXI to platelets. Analyses of missense mutations associated with FXI deficiency have provided additional clues to localization of ligand-binding sites on the protein surface. Together, these data will facilitate efforts to understand the physiology and pathology of this unusual protease, and development of therapeutics to treat thrombotic disorders.


Novel Insights on Pyoverdine: From Biosynthesis to Biotechnological Application.
Pyoverdines (PVDs) are a class of siderophores produced mostly by members of the genus Pseudomonas. Their primary function is to accumulate, mobilize, and transport iron necessary for cell metabolism. Moreover, PVDs also play a crucial role in microbes' survival by mediating biofilm formation and virulence. In this review, we reorganize the information produced in recent years regarding PVDs biosynthesis and pathogenic mechanisms, since PVDs are extremely valuable compounds. Additionally, we summarize the therapeutic applications deriving from the PVDs' use and focus on their role as therapeutic target themselves. We assess the current biotechnological applications of different sectors and evaluate the state-of-the-art technology relating to the use of synthetic biology tools for pathway engineering. Finally, we review the most recent methods and techniques capable of identifying such molecules in complex matrices for drug-discovery purposes.


The phytochemical composition and in vitro antiviral activity of decoctions from galls of Guiera senegalensis J.F. Gmel. (Combretaceae) and their relative non-toxicity for chickens.
Aqueous decoctions obtained from the galls of Guiera senegalensis were screened to determine their phytochemical composition and in vitro antiviral activity against fowlpox virus. In addition, we wanted to investigate the toxic effects, if any, of crude extracts in chickens. Steroids as well as cardiac glycosides not previously reported, an alkaloid, polyphenols and saponins were detected in the various fractions of organic solvents used for extracting the decoctions. Antiviral activity was determined by cytopathic effect inhibition assay in primary chicken embryo skin cells. The 50% inhibitory concentration (EC50) was shown to be 15.6 microg/ml. Toxicity for cells was established by determining the 50% cytotoxic concentration (CCy50). A value of 90 microg/ml and a selectivity index (CCy50/EC50) of 5.8 were obtained. In vivo studies of toxicity were performed in chickens that were dosed orally with decoctions of several concentrations for 2 weeks and then monitored for 3 months. No significant changes in several blood chemical parameters were obtained, except for a significant decline in SGOT levels in birds dosed with 100 mg/kg. These levels were nevertheless within the accepted normal range. The findings suggest that aqueous decoctions of galls from G. senegalensis are non-toxic for chickens when administered orally, even at a daily dose of 100 mg/kg for 14 days.


Do the seasonal forests in northeastern Brazil represent a single floristic unit?
Floristic analyses (Principal Component Analysis and Analysis of Group Indicators) at the genus level were employed to characterize and compare seasonal forest formations in northeastern Brazil. The presence--absence of 248 genera of woody plants occurring in 24 floristic surveys was correlated with geomorphology and climatic variables. The analyses were consistent and point to the existence of two floristic groups of seasonal forests in the region, one more closely related to the Atlantic Coastal Forest (mata atlântica) and the other to the xerophytic formations (caatinga) of the region. The driest seasonal forest group experiences more than 8 dry months per year and/or a total annual rainfall of <1000 mm, and is found on the ancient eroded peaks in the semi-arid core and on the western slopes of the Borborema Plateau.


Cellulose nanofibrils-graphene hybrids: recent advances in fabrication, properties, and applications.
With the fast-developing social economy and the acceleration of industrialization, seeking effective renewable, sustainable, and environmentally friendly resources that show promising properties is an urgent task and a crucial means to achieve sustainable progress in the face of the growing depletion of non-renewable resources and the deterioration of environmental issues. Cellulose nanofibrils (CNFs) are natural polymeric nanomaterials with excellent biocompatibility, biodegradability, good mechanical features, high strength, low density, high specific surface area, and tunable chemistry. Their combination with other nanomaterials, such as graphene derivatives (GNMs), has been demonstrated to be effective since they produce hybrids with outstanding physicochemical properties, tailorable functionality, and high performance. In this review, recent advances in the preparation, modification, and emerging application of CNFs/GNMs hybrids are described and discussed using the latest studies. First, the concise background of nanocellulose and graphene derivatives is provided, followed by the interfacial interactions between CNFs and GNMs. The different hybrids exhibit great promise in separation, adsorption, optics, flexible electronics, energy storage, thermal management, barrier and packaging, and electromagnetic shielding. The main challenges that inhibit the applicability of these hybrids are finally highlighted, and some perspectives for future research directions are provided.


Post-mortem decapitation by domestic dogs: three case reports and review of the literature.
Post-mortem animal depredation is not an uncommon phenomenon in routine forensic autopsies. We present three cases of complete post-mortem decapitation by domestic German shepherd dogs. In two cases, the head had been bitten off, defleshed and left lying near the body, while in one case it had been completely devoured by two dogs; only small skull fragments and crowned teeth could be found. Two of the three bodies were putrefied; all dog bite injuries had been inflicted after death. The cause of death was drug toxicity in two cases and fatal hemorrhage from ruptured esophageal varices in one case. These rare injuries due to post-mortem animal depredation are discussed in the light of earlier studies and case reports.


Ethical decisions in dental treatment planning using the Ozar model: a descriptive study of fifty-two consecutive patients.
Chairside dental ethics necessitates special consideration by the faculty member-dentist, the dental student, and the patient. The patient must always be aware of treatment options, in addition to learning the health status of the oral hard and soft tissues, costs, and time to be involved to accomplish proposed treatment. What may seem like a burden to an individual practitioner is actually only the necessities incurred by any self governing profession. Fifty-two consecutive dental patients had their cases reviewed for the ethical behavior of the dentist-practitioner, student, and patient. Categories reviewed were informed consent (100%), agreement, compromise, economic issues, conflict, and institutional issues. The results were: 1. Agreement: 17 cases, 2. Compromise: 21 cases, 3. Economic Issues: 4 cases, 4. Conflict: 6 cases, and 5. Institutional Issues: 4 cases. Fourteen percent of all patients were minors. Ninety percent of the patients were treated. Of those 10% not treated, five percent were for medical reasons, and five percent of the patients refused treatment. These results were placed in the Ozar Model 9 and were categorized as follows: 1. Appropriate pain free oral function: 22 cases, 2. Patient Autonomy: 14 cases, 3. Life and health: 9 cases, 4. Preferred practice values: 6 cases, and 5. Other external considerations: 1 case. This template demonstrated appropriate ethical behavior on the part of dentists, students, and patients, especially with a 90% treatment group, and only 5% refusing treatment. The 52 consecutive patients were characteristic of the University of Tennessee's overall patient pool. No identifiable differences were found between patients regardless of gender, age, or race.


Veterinary professionals' weight-related communication when discussing an overweight or obese pet with a client.
Pet weight may be difficult for veterinary professionals to address with clients, particularly when pets are overweight or obese. The objective of this study was to characterize the communication processes and content of weight-related conversations occurring between veterinary professionals and clients. Audio-video recordings of 917 veterinarian-client-patient interactions involving a random sample of 60 veterinarians and a convenience sample of clients. Companion animal veterinarians in southern Ontario, Canada, were randomly recruited, and interactions with their clients were audio-video recorded. Interactions were reviewed for mentions of weight, then further analyzed by means of a researcher-generated coding framework to provide a comprehensive assessment of communication specific to weight-related interactions. 463 of 917 (50.5%) veterinary-client-patient interactions contained an exchange involving the mention of a single patient's (dog or cat) weight and were included in final analysis. Of the 463 interactions, 150 (32.4%) involved a discussion of obesity for a single patient. Of these, 43.3% (65/150) included a weight management recommendation from the veterinary team, and 28% (42/150) provided clients with a reason for pursuing weight management. Findings illustrate opportunities to optimize obesity communication to improve the health and wellbeing of veterinary patients.


Transapical aortic valve implantation with a self-expanding anatomically oriented valve.
The Medtronic Engager™ aortic valve bioprosthesis is a self-expanding valve with support arms facilitating anatomically correct positioning and axial fixation. Valve leaflets, made of bovine pericardium, are mounted on a Nitinol frame. Here, we report the first in man study with this new implant (Trial Identifier NCT00677638). Thirty patients (mean age 83.4 ± 3.8 years; 83% female) with tricuspid aortic valve stenosis were included in the study. Mean logistic EuroSCORE was 23.4 ± 11.9. Mean aortic annulus diameter was 21.8 ± 1.4 mm. For this study, the Engager was available in only one size (23 mm), to fit aortic annuli of 19-23 mm. Standard transapical valve implantation was performed using pre-dilation of the aortic valve and rapid ventricular pacing during balloon valvuloplasty and most valve deployments. Accurate valve placement was achieved in 29/30 cases (97%). Post-implant peak-to-peak gradient was 13.3 ± 9.3 mmHg. In 80% of the patients, no more than grade I paravalvular leakage was observed, in 13% grades I-II and in 3% grade II. Three patients (10%) required permanent pacemaker implantation for higher-degree or complete atrioventricular block. Four dissections (13%) occurred during positioning of the valve and were treated surgically in three cases. Thirty-day and in-hospital mortality were 20% and 23%, respectively, and 6-month survival was 56.7%. No structural failure occurred for up to 1 year. This series established the feasibility of implanting a novel self-expanding transapical aortic valve prosthesis predictably into an anatomically correct position. Observed complications led to complete redesign of the delivery system for upcoming clinical studies with the goal of establishing safety and performance.


Binding of Nicotiana nuclear proteins to the subterminal regions of the Ac transposable element.
Specific binding of Nicotiana nuclear protein(s) to subterminal regions of the Ac transposable element was detected using gel mobility shift assays. A sequence motif (GGTAAA) repeated in both terminal regions of Ac, was identified as the protein binding site. Mutation of two nucleotides in this motif was sufficient to abolish binding. Based on a series of competition assays, it is deduced that there is cooperative binding between two repeats, each similar to the GGTAAA motif. The binding protein is probably similar to a previously characterized maize protein which binds to a GGTAAA-containing motif located in the ends of Mutator. Moreover, we show that DNA from Ds1 competes for protein binding to Ac termini, and we show, by sequence analysis, that GGTAAA binding sites are present in the terminal region of Tgm1, Tpn1, En/Spm, Tam3, and Ds1-like elements. This suggests that the binding protein(s) might be involved in the transposition process.


Physical activity, physical frailty and depressive symptoms among Chinese male and female older adults: do different indicators make a difference?
Older adults become more inactive and frailer with aging. Physical status is closely linked to mental health, but it is unclear which physical indicator is more strongly associated with depressive symptoms in older adults. The present study aimed to compare relationships between self-reported physical activity, physical frailty (muscle mass, muscle strength, and gait ability) and depressive symptoms in community male and female older adults. A total of 1,180 adults aged 60 years and older were recruited to participate in this study from a Chinese community receiving annual check-up service from September 2018 to May 2019. Physical activity was assessed by the International Physical Activity Questionnaire (IPAQ). The Bio-electrical Impedance Analyzer was used to determine the muscle mass. As the indicators of muscle function, grip strength and gait ability were assessed by the dynamometer and Timed Up and Go Test (TUGT), respectively. The 15-item version of Geriatric Depression Scale (GDS-15) was used to examine depressive symptoms. Demographic variables, health status and sleep quality were collected using questionnaire. 11.8% men and 11.9% women reported depressive symptoms. Logistic regression showed that depressive symptoms was associated with low grip strength (OR = 2.42, 95% CI: 1.04-5.63), slow gait ability (OR = 3.60, 95% CI: 1.28-10.13) in older males, and associated with low level of self-reported physical activity (OR = 3.85, 95% CI: 2.00-7.42) in older females. No significant association was found between muscle mass and depressive symptoms. There were gender differences in the relationship between physical activity, physical frailty, and depressive symptoms. Grip strength and gait ability may be a better indicator of frailty for predicting depressive symptoms in older men while physical activity may be useful in predicting depressive symptoms in older women.


Validating the Riverside Acculturation Stress Inventory with Asian Americans.
An emerging body of empirical research highlights the impact of acculturative stress in the lives of culturally diverse populations. Therefore, to facilitate future research in this area, we conducted 3 studies to examine the psychometric properties of the Riverside Acculturation Stress Inventory (RASI; Benet-Martínez & Haritatos, 2005) and its 5 subscales in a total sample of 793 self-identified Asian American participants. The reliability and validity of RASI scores and the hypothesized 1-factor higher order model (with 1st-order factors Language Skills, Work Challenges, Intercultural Relations, Discrimination, and Cultural Isolation) of the RASI were examined in Study 1. The RASI higher order structure and score validity and reliability were examined across different generational groups in Study 2. The stability of RASI scores over a 3-week period was examined in Study 3. Overall, findings from these studies support the hypothesized structure of the RASI and indicate that this brief instrument provides reliable and valid acculturative stress scores. In addition, results suggest that RASI items are interpreted in an equivalent manner across different generations of Asian American individuals. Implications for research and assessment are discussed.


Radiofrequency radiation injures trees around mobile phone base stations.
In the last two decades, the deployment of phone masts around the world has taken place and, for many years, there has been a discussion in the scientific community about the possible environmental impact from mobile phone base stations. Trees have several advantages over animals as experimental subjects and the aim of this study was to verify whether there is a connection between unusual (generally unilateral) tree damage and radiofrequency exposure. To achieve this, a detailed long-term (2006-2015) field monitoring study was performed in the cities of Bamberg and Hallstadt (Germany). During monitoring, observations and photographic recordings of unusual or unexplainable tree damage were taken, alongside the measurement of electromagnetic radiation. In 2015 measurements of RF-EMF (Radiofrequency Electromagnetic Fields) were carried out. A polygon spanning both cities was chosen as the study site, where 144 measurements of the radiofrequency of electromagnetic fields were taken at a height of 1.5m in streets and parks at different locations. By interpolation of the 144 measurement points, we were able to compile an electromagnetic map of the power flux density in Bamberg and Hallstadt. We selected 60 damaged trees, in addition to 30 randomly selected trees and 30 trees in low radiation areas (n=120) in this polygon. The measurements of all trees revealed significant differences between the damaged side facing a phone mast and the opposite side, as well as differences between the exposed side of damaged trees and all other groups of trees in both sides. Thus, we found that side differences in measured values of power flux density corresponded to side differences in damage. The 30 selected trees in low radiation areas (no visual contact to any phone mast and power flux density under 50μW/m2) showed no damage. Statistical analysis demonstrated that electromagnetic radiation from mobile phone masts is harmful for trees. These results are consistent with the fact that damage afflicted on trees by mobile phone towers usually start on one side, extending to the whole tree over time.


Immunohistochemical Expression of Angiotensin-Converting Enzyme 2 in the Skin of Patients Affected by COVID-19.
After many recorded cases of acute pneumonia of unknown cause, the World Health Organization announced COVID-19 as the start of a new coronavirus disease pandemic in 2019. Angiotensin-converting enzyme-2 (ACE2) is reduced by a protease known as transmembrane serine type 2 in the host cell, which then activates the S protein of SARS-CoV-2 regulating coronavirus entry into the host cells. The aim of this study was to assess the immunohistochemical expression of ACE 2 in the skin of patients affected by COVID-19 with and without cutaneous manifestations and to correlate ACE2 expression with clinical and pathologic parameters. Skin biopsies were obtained from skin lesions of 25 patients presenting with cutaneous manifestations and from the left forearm of 22 patients without cutaneous manifestations. The specimens were processed for evaluation of histopathologic changes and ACE2 immunohistochemical evaluation. Positive ACE2 expression was significantly higher in patients without cutaneous manifestations (96%) than those with cutaneous manifestations (72.7%). Positive ACE2 expression in the skin of affected patients was significantly associated with the presence of comorbidities, positive family history, high ABCD score, elevated lactate dehydrogenase, high D-dimer, rapid respiratory rate, and low oxygen saturation. The skin could be involved in COVID-19 infection in the form of inflammatory changes, such as pityriasis rosea-like lesions. Patients with COVID-19 who presented with cutaneous manifestations are usually less severe. The presence of ACE2 in the skin of patients with COVID-19 is an indicator of worse status. Patients with COVID-19 without skin manifestations showed higher positivity for ACE2, which may explain the severity of the cases.


Diet Quality Index associated with Digital Food Guide: update and validation.
Monitoring and assessing dietary intake may favor the promotion of healthy choices and the indexes that assess the quality of the diet contribute to this purpose. The Diet Quality Index associated with the Digital Food Guide (DQI-DFG) was developed from guidelines of the School of Public Health at Harvard University, adjusted for the Brazilian food habits. Based on new studies on prevention of chronic non-communicable diseases, the objective of this study was to improve and validate the DQI-DFG for a second version. The following psychometric properties were used: (a) content validation: consensus on healthy eating and DQI among experts; (b) construct validation: enhancement of a reference diet to determine recommended serving sizes and consumption ranges; correlation between each group score and energy value; comparative analyses between menu assessment by experts and DQI-DGF results to confirm the score criterion; (c) reliability: agreement between the result shown by the evaluators and the result presented by DQI-DFG; correlation between each food group score and total DQI score to understand the relationship between these variables. The energetic values do not influence the DQI score. There is a high correlation between the score attributed by experts and the result of DQI-DFG (r = 0.78 until r = 0.97). The Index components have a balanced score of influence in its final result (r = 0.49 until r = 0.10). Consensus result performed among experts legitimates the concepts that justify DQI-DFG. There was an agreement between menu quality evaluation by experts and the results shown by DQI-DFG.


Effects of immunization with LDH-X on fertility.
Immunological regulation of fertility can be best achieved if a defined macromolecule is used to provide the antigenic stimulus. Proteins with known enzymatic properties are attractive molecules in this area if it can be demonstrated unequivocally that they are unique constituents of spermatozoa, and are completely foreign to the female. This paper describes immunosuppression of fertility by the sperm-specific isozyme of lactate dehydrogenase (LDH-X). In addition, evidence is reviewed which suggests that acrosomal proteinase and hyaluronidase may be unique to sperm. Data are presented which confirm the sperm specificity of LDH-X and the immunochemical homogeneity of purified preparations of this isozyme. Immunization of female rabbits with LDH-X significantly reduces the fertility of these animals. Experimental results indicate that the primary effect of immunization involves blockage of fertilization. While those ova which are fertilized in an immune environment develop and implant normally, there is a high rate of post-implantation embryo mortality. Rabbit blastocysts transferred to immune recipients implant at the same rate as in non-immune controls. These findings are consistent with the conclusion that the developing embryo must interact with antibody during the pre-implantation stages of pregnancy when antigenic sites conferred by the sperm LDH-X are available. Such interaction can be demonstrated directly by immunofluorescence techniques.


Characterization of the peroxidase mechanism upon reaction of prostacyclin synthase with peracetic acid. Identification of a tyrosyl radical intermediate.
Prostacyclin synthase (PGIS) is a membrane-bound class III cytochrome P450 that catalyzes an isomerization of prostaglandin H(2), an endoperoxide, to prostacyclin. We report here the characterization of the PGIS intermediates in reactions with other peroxides, peracetic acid (PA), and iodosylbenzene. Rapid-scan stopped-flow experiments revealed an intermediate with an absorption spectrum similar to that of compound ES (Cpd ES), which is an oxo-ferryl (Fe(IV)O) plus a protein-derived radical. Cpd ES, formed upon reaction with PA, has an X-band (9 GHz) EPR signal of g = 2.0047 and a half-saturation power, P(1/2), of 0.73 mW. High-field (130 GHz) EPR reveals the presence of two species of tyrosyl radicals in Cpd ES with their g-tensor components (g(x), g(y), g(z)) of 2.00970, 2.00433, 2.00211 and 2.00700, 2.00433, 2.00211 at a 1:2 ratio, indicating that one is involved in hydrogen bonding and the other is not. The line width of the g = 2 signal becomes narrower, while its P(1/2) value becomes smaller as the reaction proceeds, indicating migration of the unpaired electron to an alternative site. The rate of electron migration ( approximately 0.2 s(-1)) is similar to that of heme bleaching, suggesting the migration is associated with the enzymatic inactivation. Moreover, a g = 6 signal that is presumably a high-spin ferric species emerges after the appearance of the amino acid radical and subsequently decays at a rate comparable to that of enzymatic inactivation. This loss of the g = 6 species thus likely indicates another pathway leading to enzymatic inactivation. The inactivation, however, was prevented by the exogenous reductant guaiacol. The studies of PGIS with PA described herein provide a mechanistic model of a peroxidase reaction catalyzed by the class III cytochromes P450.


Body weight supported treadmill training in acute spinal cord injury: impact on muscle and bone.
Longitudinal prospective case series. To evaluate the impact of early introduction post-spinal cord injury (SCI) of twice-weekly body-weight supported treadmill training (BWSTT) on muscle and bone. Centre for Health Promotion and Rehabilitation, McMaster University, Canada. Five individuals who had sustained traumatic SCI within 2-6 months participated in the study. Bone mineral densities (BMD) of proximal femur, distal femur, proximal tibia and lumbar spine were measured before and after training, as well as muscle cross-sectional area (CSA), BMD and bone geometry at mid-femur and proximal tibia. Serum osteocalcin and urinary deoxypyridinoline were measured at baseline, and after 24 and 48 sessions of training. All participants experienced increased muscle CSAs, ranging from 3.8 to 56.9%. Reductions in BMD were evident in all participants at almost all lower limb sites after training, ranging in magnitude from -1.2 to -26.7%. Lumbar spine BMD changes ranged from 0.2 to -7.4%. No consistent changes were observed in bone geometry. BWSTT did not alter the expected pattern of change in bone biochemical markers over time. The individual with the greatest improvement in ambulatory ability demonstrated the smallest reduction in lower limb BMD. Conversely, the individual who completed the fewest BWSTT sessions demonstrated the greatest reductions in BMD. Twice-weekly BWSTT appeared to partially reverse muscle atrophy after SCI, but did not prevent bone loss. Larger, controlled trials should evaluate whether relative preservation of bone loss occurs with regular BWSTT following acute SCI. Ontario Neurotrauma Foundation.


Effects of germinated barley foodstuff on dextran sulfate sodium-induced colitis in rats.
Germinated barley foodstuff (GBF), derived from the aleurone and scutellum fractions of germinated barley, is rich in glutamine and low-lignified hemicellulose, and increases mucosal protein, RNA, and DNA content in the intestine when fed to normal rats. The aim of this study was to evaluate the effects of feeding GBF or germinated gramineous seeds on experimental ulcerative colitis. Sprague-Dawley rats that received 3% dextran sulfate sodium in their diets were used as an experimental colitis model. The effects of sulfasalazine, a drug used to treat inflammatory bowel disease, were compared with those of GBF. After rats had consumed diets containing GBF or various aleurone and scutellum fractions, mucosal damage; the content of mucosal protein, RNA, and DNA in the colo-rectum; and serum interleukin-8 and alpha1-acid glycoprotein levels were assessed. GBF and germinated seeds more effectively prevented bloody diarrhea and mucosal damage in colitis compared with controls and rats receiving sulfasalazine, but non-germinated samples did not have a protective effect. GBF increased mucosal protein and RNA content in the colitis model. The consumption of GBF appears to prevent inflammation in a colitis model, and its effect seems to be related to the germination process. GBF and germinated seeds have the potential to serve as nutritional therapy for ulcerative colitis.


L-serine binds to arginine-148 of the beta 2 subunit of Escherichia coli tryptophan synthase.
Inactivation of the beta 2 subunit and of the alpha 2 beta 2 complex of tryptophan synthase of Escherichia coli by the arginine-specific dicarbonyl reagent phenylglyoxal results from modification of one arginyl residue per beta monomer. The substrate L-serine protects the holo beta 2 subunit and the holo alpha 2 beta 2 complex from both inactivation and arginine modification but has no effect on the inactivation or modification of the apo forms of the enzyme. This result and the finding that phenylglyoxal competes with L-serine in reactions catalyzed by both the holo beta 2 subunit and the holo alpha 2 beta 2 complex indicate that L-serine and phenylglyoxal both bind to the same essential arginyl residue in the holo beta 2 subunit. The apo beta 2 subunit is protected from phenylglyoxal inactivation much more effectively by phosphopyridoxyl-L-serine than by either pyridoxal phosphate or pyridoxine phosphate, both of which lack the L-serine moiety. The phenylglyoxal-modified apo beta 2 subunit binds pyridoxal phosphate and the alpha subunit but cannot bind L-serine or L-tryptophan. We conclude that the alpha-carboxyl group of L-serine and not the phosphate of pyridoxal phosphate binds to the essential arginyl residue in the beta 2 subunit. The specific arginyl residue in the beta 2 subunit which is protected by L-serine from modification by phenyl[2-14C]glyoxal has been identified as arginine-148 by isolating a labeled cyanogen bromide fragment (residues 135-149) and by digesting this fragment with pepsin to yield the labeled dipeptide arginine-methionine (residues 148-149). The primary sequence near arginine-148 contains three other basic residues (lysine-137, arginine-141, and arginine-150) which may facilitate anion binding and increase the reactivity of arginine-148. The conservation of the arginine residues 141, 148, and 150 in the sequences of tryptophan synthase from E. coli, Salmonella typhimurium, and yeast supports a functional role for these three residues in anion binding. The location and role of the active-site arginyl residues in the beta 2 subunit and in two other enzymes which contain pyridoxal phosphate, aspartate aminotransferase and glycogen phosphorylase, are compared.


Fractures of the calcaneum. A review of 70 patients.
We have assessed the long-term results after operative and non-operative treatment of undisplaced and displaced calcaneal fractures. At a mean of 6.5 years, we reviewed 70 patients with a calcaneal fracture who were divided into four groups: group 1, 18 patients with undisplaced fractures and a normal Böhler's angle (BA) who had been treated non-operatively; group 2, 23 with intra-articular fractures and a BA < 10 degrees who had been treated non-operatively; group 3, 13 with intra-articular fractures and a BA > 10 degrees who had been treated surgically; and group 4, 16 with intra-articular fractures and a BA < 10 degrees who had been treated surgically. The results were assessed by a clinical score considering pain, return to work, return to physical activity, change in shoe-wear and the requirement for subtalar arthrodesis. Patients with undisplaced calcaneal fractures had a good outcome. Those with displaced fractures treated surgically who presented at follow-up with a BA > 10 degrees had a satisfactory functional outcome and those with displaced fractures who had non-operative treatment had a poor outcome. The poorest outcome was consistently seen in patients who were treated operatively without restoration of BA. Open reduction and internal fixation of intra-articular calcaneal fractures can only be expected to benefit those patients in whom nearly anatomical reconstruction is obtained.


Infantile granular parakeratosis: cytologic examination of superficial scrapings as an aid to diagnosis.
Granular parakeratosis (GP) is a benign disorder of keratinization presenting with unilateral or bilateral red to brown hyperkeratotic plaques and papules at the intertriginous areas. The first pediatric case of GP was reported in 2002, and only a few cases have been reported since. Although the exact etiology of GP is unknown, it is thought that certain environmental factors compromise the epidermal barrier and lead to proliferation and altered maturation of the epidermis in predisposed individuals. The histopathology is diagnostic and reveals parakeratosis together with retention of keratohyalin granules within a disproportionately thickened stratum corneum and usually the preservation of stratum granulosum. Herein we report seven cases of infantile GP, six of which also had atopic dermatitis. Cytologic examination confirmed our clinical diagnosis by demonstrating the retention of keratohyalin granules and preservation of the nuclei from the superficial scrapings of the lesions, which we propose as a novel diagnostic technique. In all seven cases the lesions developed after the overuse of topical products and resolved with avoidance of their excessive use. We propose that atopic skin may be more prone to develop GP because the epidermal barrier is disrupted, resulting in the enhanced transepidermal penetration of topical products. In conclusion, GP should be included in the differential diagnosis of diaper area eruptions, especially in atopic children. Cytologic examination of superficial scrapings of the lesions can easily confirm the diagnosis of GP.


Being Thankful: Examining the Relationship Between Young Adult Religiosity and Gratitude.
Studies show that religious people tend to be more grateful, yet existing research tends to rely on small, non-representative samples and limited measures of religiosity. Therefore, we use a national sample (the National Study of Youth and Religion) and multiple measures of religiosity to examine the extent to which religion is associated with feelings of gratitude. We find that religious efficacy and having religious friends positively predict the extent to which young adults feel grateful. In contrast, religious affiliation, participation in organized religion, private devotion, religious salience, otherworldly beliefs, and being spiritual but not religious are unrelated to experiencing feelings of gratitude.


The sarcoplasmic reticulum: Actin and tropomodulin hit the links.
Skeletal muscle exhibits strikingly regular intracellular sorting of actin and tropomodulin (Tmod) isoforms, which are essential for efficient muscle contraction. A recent study from our laboratory demonstrates that the skeletal muscle sarcoplasmic reticulum (SR) is associated with cytoplasmic γ-actin (γ(cyto)-actin) filaments, which are predominantly capped by Tmod3. When Tmod3 is experimentally induced to vacate its SR compartment, the cytoskeletal organization of SR-associated γ(cyto)-actin is perturbed, leading to SR swelling, depressed SR Ca(2+) release and myofibril misalignment. Based on these findings, Tmod3-capped γ(cyto)-actin filaments mechanically stabilize SR structure and regulate SR function via a novel lateral linkage. Furthermore, by placing these findings in the context of studies in nonmuscle cells, we conclude that Tmodcapped actin filaments are emerging as critical regulators of membrane stability and physiology in a broad assortment of cell types.


Assessing Impressions of Community Health Worker Use of Tablet-Based Medication Documentation Software.
Community health workers, an important and emerging role in the evolution of the care delivery system, are uniquely positioned to play a vital role in information gathering and exchange. NewYork-Presbyterian Hospital carried out a feasibility study to assess community health worker use and impressions of a medication documentation application that runs on a tablet. The nine community health workers successfully collected data on 16 patients in the home setting. On average, 10 medications were collected per patient, and the average time to collect the medication data was 1 hour. Key findings from the focus groups included the need for additional training and the desire to use the device to streamline other documentation activities. In general, the software was judged to be suitable for the proposed task and represents a starting point for further use of tools that leverage the community health worker in a team-based workflow.


HYDROXYUREA: INHIBITORY EFFECT ON DNA METABOLISM.
Hydroxyurea, hydroxyurethane, and dihydroxyurea inhibit incorporation of thymidine into the DNA of monolayers of HeLa cells. They do not affect incorporation of uridine into RNA or of leucine into protein. In contrast, hydroxylamine inhibits cellular incorporation of all three precursors: thymidine, uridine, and leucine. Hydroxyurea does not affect thymidine kinase, thymidylate kinase, or DNA polymerase reactions, but it does inhibit incorporation of cytidylic and guanylic acids into DNA in cell-free supernatants.


Bulk ionic screening lengths from extremely large-scale molecular dynamics simulations.
Recent experiments have reported anomalously large screening lengths of interactions between charged surfaces confining concentrated electrolytes and ionic liquids. Termed underscreening, this effect was ascribed to bulk properties of dense ionic systems. Herein, we study bulk ionic screening with extremely large-scale molecular dynamics simulations, allowing us to assess the range of distances relevant to the experiments. Our results yield two screening lengths satisfying distinct scaling relations. However, with an accuracy of 10-5kBT in interionic potentials of mean force, we find no signs of underscreening, suggesting that other than bulk effects might be at play in the experiments.


[Heat Shock Proteins in Plant Protection from Oxidative Stress].
This review considers the recent progress on the role of heat shock proteins (HSPs), as well as transcription factors of heat shock proteins genes (HSFs) in protecting plants from oxidative stress induced by various types of abiotic and biotic stresses. HSPs are pleiotropic proteins involved in various intracellular processes and performing many important functions. In particular, HSPs increase plant resistance to stress by protecting the structure and activity of proteins of the antioxidant system. Overexpression of Hsp genes under stressful conditions, leading to an increased content of HSPs, can be used as a marker of oxidative stress. Plant HSFs are encoded by large gene families with variable sequences, expression and function. Plant HSFs regulate transcription of a wide range of stress-induced genes, including HSPs and other chaperones, reactive oxygen species scavengers, enzymes involved in protective metabolic reactions and osmolytic biosynthesis, or other transcriptional factors. Genome-wide analysis of Arabidopsis, rice, poplar, lettuce, and wheat revealed a complex network of interaction between the Hsps and Hsfs gene families that form plant protection against oxidative stress. Plant protection systems are discussed, with special emphasis on the role of HSPs and HSFs in plant responses to stress, which will be useful for the development of technologies to increase productivity and stress resistance of plant crops.


Cis-trans isomers of vitamin A and retinene in the rhodopsin system.
Vitamin A and retinene, the carotenoid precursors of rhodopsin, occur in a variety of molecular shapes, cis-trans isomers of one another. For the synthesis of rhodopsin a specific cis isomer of vitamin A is needed. Ordinary crystalline vitamin A, as also the commercial synthetic product, both primarily all-trans, are ineffective. The main site of isomer specificity is the coupling of retinene with opsin. It is this reaction that requires a specific cis isomer of retinene. The oxidation of vitamin A to retinene by the alcohol dehydrogenase-cozymase system displays only a low degree of isomer specificity. Five isomers of retinene have been isolated in crystalline condition: all-trans; three apparently mono-cis forms, neoretinenes a and b and isoretinene a; and one apparently di-cis isomer, isoretinene b. Neoretinenes a and b were first isolated in our laboratory, and isoretinenes a and b in the Organic Research Laboratory of Distillation Products Industries. Each of these substances is converted to an equilibrium mixture of stereoisomers on simple exposure to light. For this reaction, light is required which retinene can absorb; i.e., blue, violet, or ultraviolet light. Yellow, orange, or red light has little effect. The single geometrical isomers of retinene must therefore be protected from low wave length radiation if their isomerization is to be avoided. By incubation with opsin in the dark, the capacity of each of the retinene isomers to synthesize rhodopsin was examined. All-trans retinene and neoretinene a are inactive. Neoretinene b yields rhodopsin indistinguishable from that extracted from the dark-adapted retina (lambda(max.) 500 mmicro). Isoretinene a yields a similar light-sensitive pigment, isorhodopsin, the absorption spectrum of which is displaced toward shorter wave lengths (lambda(max.) 487 mmicro). Isoretinene b appears to be inactive, but isomerizes preferentially to isoretinene a, which in the presence of opsin is removed to form isorhodopsin before the isomerization can go further. The synthesis of rhodopsin in solution follows the course of a bimolecular reaction, as though one molecule of neoretinene b combines with one of opsin. The synthesis of isorhodopsin displays similar kinetics. The bleaching of rhodopsin, whether by chemical means or by exposure to yellow or orange (i.e., non-isomerizing) light, yields primarily or exclusively all-trans retinene. The same appears to be true of isorhodopsin. The process of bleaching is therefore intrinsically irreversible. The all-trans retinene which results must be isomerized to active configurations before rhodopsin or isorhodopsin can be regenerated. A cycle of isomerization is therefore an integral part of the rhodopsin system. The all-trans retinene which emerges from the bleaching of rhodopsin must be isomerized to neoretinene b before it can go back; or if first reduced to all-trans vitamin A, this must be isomerized to neovitamin Ab before it can regenerate rhodopsin. The retina obtains new supplies of the neo-b isomer: (a) by the isomerization of all-trans retinene in the eye by blue or violet light; (b) by exchanging all-trans vitamin A for new neovitamin Ab from the blood circulation; and (c) the eye tissues may contain enzymes which catalyze the isomerization of retinene and vitamin A in situ. When the all-trans retinene which results from bleaching rhodopsin in orange or yellow light is exposed to blue or violet light, its isomerization is accompanied by a fall in extinction and a shift of absorption spectrum about 5 mmicro toward shorter wave lengths. This is a second photochemical step in the bleaching of rhodopsin. It converts the inactive, all-trans isomer of retinene into a mixture of isomers, from which mixtures of rhodopsin and isorhodopsin can be regenerated. Isorhodopsin, however, is an artefact. There is no evidence that it occurs in the retina; nor has isovitamin Aa or b yet been identified in vivo. In rhodopsin and isorhodopsin, the prosthetic groups appear to retain the cis configurations characteristic of their retinene precursors. In accord with this view, the beta-bands in the absorption spectra of both pigments appear to be cis peaks. The conversion to the all-trans configuration occurs during the process of bleaching. The possibility is discussed that rhodopsin may represent a halochromic complex of a retinyl ion with opsin. The increased resonance associated with the ionic state of retinene might then be responsible both for the color of rhodopsin and for the tendency of retinene to assume the all-trans configuration on its release from the complex. A distinction must be made between the immediate precursor of rhodopsin, neovitamin Ab, and the vitamin A which must be fed in order that rhodopsin be synthesized in vivo. Since vitamin A isomerizes in the body, it is probable that any geometrical isomer can fulfill all the nutritional needs for this vitamin.


Effect of trace iron levels and iron withdrawal by chelation on the growth of Candida albicans and Candida vini.
The iron requirements of the opportunistic pathogenic yeast, Candida albicans, and the related nonpathogenic spoilage yeast Candida vini were investigated along with their responses to various exogenous iron chelators. The influence of iron as well as the exogenous chelating agents lactoferrin, EDTA, deferiprone, desferrioxamine, bathophenanthroline sulphonate and a novel carried chelator with a hydroxypyridinone-like Fe-ligand functionality, DIBI, on fungal growth was studied in a chemically defined medium deferrated to trace iron levels (<1.2 microg L(-1) or 0.02 microM of Fe). Candida albicans competed better at low iron levels compared with C. vini, which was also more susceptible to most added chelators. Candida albicans was resistant to lactoferrin at physiologically relevant concentrations, but was inhibited by low concentrations of DIBI. Candida vini was sensitive to lactoferrin as well as to DIBI, whose inhibitory activity was shown to be Fe reversible. The pathogenic potential of C. albicans and the nonpathogenic nature of C. vini were consistent with their differing abilities to grow under iron-limiting conditions and in the presence of exogenous iron chelators. Both yeasts could be controlled by appropriately strong chelators. This work provides the first evidence of the iron requirements of the spoilage organism C. vini and its response to exogenous chelators. Efficient iron withdrawal has the potential to provide the basis for new fungal growth control strategies.


Failure to keep clinic appointments: implications for residency education and productivity.
Missed clinic appointments significantly impact continuity of care, resident education, and clinical productivity. This national survey analyzed the awareness of family practice residency clinics regarding their missed appointment rate and the use of reminder systems. We mailed a questionnaire to all 468 family practice residency programs in the United States and mailed two follow-up surveys to nonrespondents. The overall survey response rate was 60%. More than one third of the clinics reported a no-show rate of > 21%. When asked how the clinic arrived at their no-show rate, 64 (17.7%) respondents stated that the reported rate was an estimate. More than one third of all clinics did not use a patient reminder system for clinic appointments. When comparing clinics with high (> 20%) and low (< 20%) no-show rates, there was no statistically significant difference between the use or nonuse of mailed, telephone, or combined reminder systems. The clinic appointment no-show rate varies greatly among family practice residency programs. Even with the widely reported use of reminder systems, one third of programs continue to have no-show rates above 20%. This study is unusual in that there was no statistically significant difference in no-show rates between programs, based on the use of reminder systems. In an era of cost containment and increased accountability, appointment failures are a factor that will require additional study and intervention by family practice residency programs nationwide.


Molecular diagnosis of viral disease in hematology patients.
Viruses, through their direct and indirect effects on hematopoietic cells, are not only etiological agents of hematologic disease, but they both complicate transfusion therapy and are major causes of morbidity and mortality in immunosuppressed patients. The development of molecular techniques, especially DNA hybridization and amplification, has allowed new viruses to be discovered, has aided in the early diagnosis of infection before the development of an antibody response, and has revolutionized the detection of viruses in blood transfusions.


What do expressions with north and northern people tell us?
The aim of this article is to study linguistic expressions which include the words North and Northern people. This is done by philological and linguistic analysis of the material, which consists of different types of written documents dating from different historical periods. The oldest historical documents were produced by the Ancient Roman writers, who wrote about wild tribes living in the North. The expression "the evil from the North" in the Bible is also used by the Fathers of the Church, when describing the Great Migrations. The most recent material consists of academic texts-dealing with the Northern dimension--and everyday texts--in this case, mostly of bread packages--referring among other things to people's nutrition and health. Expressions referring to the North are also present in modern material, but the negative association of the older documents is not so present. All the texts, both historical and modern, are discussed as a whole from Finnish and Scandinavian points of view. Semiotic aspects are highlighted by the use of some visual illustrations from the different texts in point.


Therapeutic drug monitoring of the β-lactam antibiotics: what is the evidence and which patients should we be using it for?
Traditional antibiotic dosing was not designed for today's escalating antibiotic resistance, lack of novel antibiotics and growing complexity in patient populations. Dosing that ensures optimal antibiotic exposures should be considered essential to increase the likelihood of effective patient treatment. Given the variability in these exposures across different patients, a 'one-dose-fits-all' approach is increasingly problematic. Therapeutic drug monitoring (TDM) of the β-lactams, the most widely used antibiotic class, is underutilized in certain populations. Clinical experience with β-lactam TDM remains relatively scarce. Patients most likely to benefit from such an intervention include the critically ill, the obese, the elderly and those with cystic fibrosis. Most centres actively performing β-lactam TDM target a minimum 100% of the time during the dosing interval that the free (unbound) concentration of antibiotic exceeds the MIC of the pathogen (100% fT>MIC), which is higher than a traditional target supported by in vitro data. Ideally, isolated pathogens should undergo MIC testing along with TDM on a regular basis, allowing clinicians to address the triad of bug, drug and patient ('mug') in equal measure.


Isolation and characterization of ZZ1, a novel lytic phage that infects Acinetobacter baumannii clinical isolates.
Acinetobacter baumannii, a significant nosocomial pathogen, has evolved resistance to almost all conventional antimicrobial drugs. Bacteriophage therapy is a potential alternative treatment for multidrug-resistant bacterial infections. In this study, one lytic bacteriophage, ZZ1, which infects A. baumannii and has a broad host range, was selected for characterization. Phage ZZ1 and 3 of its natural hosts, A. baumanni clinical isolates AB09V, AB0902, and AB0901, are described in this study. The 3 strains have different sensitivities to ZZ1, but they have the same sensitivity to antibiotics. They are resistant to almost all of the antibiotics tested, except for polymyxin. Several aspects of the life cycle of ZZ1 were investigated using the sensitive strain AB09V under optimal growth conditions. ZZ1 is highly infectious with a short latent period (9 min) and a large burst size (200 PFU/cell). It exhibited the most powerful antibacterial activity at temperatures ranging from 35°C to 39°C. Moreover, when ZZ1 alone was incubated at different pHs and different temperatures, the phage was stable over a wide pH range (4 to 9) and at extreme temperatures (between 50°C and 60°C). ZZ1 possesses a 100-nm icosahedral head containing double-stranded DNA with a total length of 166,682 bp and a 120-nm long contractile tail. Morphologically, it could be classified as a member of the Myoviridae family and the Caudovirales order. Bioinformatic analysis of the phage whole genome sequence further suggested that ZZ1 was more likely to be a new member of the Myoviridae phages. Most of the predicted ORFs of the phage were similar to the predicted ORFs from other Acinetobacter phages. The phage ZZ1 has a relatively broad lytic spectrum, high pH stability, strong heat resistance, and efficient antibacterial potential at body temperature. These characteristics greatly increase the utility of this phage as an antibacterial agent; thus, it should be further investigated.


Effects of silver nanoparticles on biological nitrogen removal processes.
The effects of silver (Ag) nanoparticles (NPs) on activated sludge in a biological nitrogen removal (BNR) process were investigated under aerobic and anoxic conditions. We show that nitrification was more vulnerable to Ag NPs exposure than denitrification at the same Ag NPs concentration. In continuous operation of the BNR process, a higher inhibitory effect on nitrification was attributed to a smaller size of Ag NPs. About 70-90% of the Ag NPs supplied were embedded in the sludge matrix but 10-30% of the Ag NPs remained in the supernatant. This indicates that significant amounts of Ag NPs could be discharged from wastewater treatment plants and potentially impact on aquatic ecosystems.


Estimating the causal effect of compliance on binary outcome in randomized controlled trials.
We examine likelihood based methods aimed at analysing the causal effect of actual exposure to drug treatment on a (repeated) binary outcome in two randomized trials with partial compliance. Starting with the univariate compliance summary 'total treatment dose history', we apply a method for ordinal compliance and monotone dose response, proposed by Goetghebeur and Molenberghs. In a short duration trial of blood pressure reduction, this summary leads to meaningful effect estimators. However, in the analysis of a vitamin A trial, this method reaches a boundary solution; the estimated possible benefit from vitamin A for children who did not receive any pills on the treatment arm is zero. In our formulation the number of pills that were taken captures part of the outcome, and the corresponding effect parameters suffer from this confounding. To gain additional insight, we account explicitly for the temporal structure of compliance. We extend the likelihood based methodology for univariate ordered compliance to more dimensional compliance with only a partial order structure on exposure. The randomization assumptions in the causal formulation of Rubin are translated to this setting. We motivate a set of parametric assumptions on the joint distribution of potential outcomes and observed compliance levels and reanalyse the vitamin A trial. Our findings suggest that one capsule of vitamin A had a large impact on mortality during the first 4 months. The greatest reduction in risk was estimated amongst children who received two doses. This supports findings from a vitamin A trial in Ghana and in Nepal. Finally, we discuss extensions of this method, covering uncensored and censored grouped survival data.


EEG Source Imaging of Infarct Core and Penumbra for Ischemic Stroke Patients.
A quantitative method of analyzing EEG signals after stroke onset can help monitor disease progression and tailor treatments. In this work, we present an EEG-based imaging algorithm to estimate the location and size of the stroke infarct core and penumbra tissues. Building on recent advancements in localizing neural silences, we develop an algorithm that utilizes known spectral properties of the infarct core and penumbra to separately localize them. Our algorithm uses these properties to estimate source contributions to the scalp EEG recordings in different frequency bands. Subsequently, it utilizes optimization techniques to search for the affected brain sources iteratively. We test our algorithm on simulated datasets using a realistic MRI head model, achieving center-of-mass error of 12.80mm and 17.24mm, and size estimation error of 21.78% and 36.62% for infarct core and penumbra respectively.


Cost-utility analysis of salvage cardiac extracorporeal membrane oxygenation in children.
Indications for extracorporeal membrane oxygenation therapy have expanded to include cardiopulmonary arrest and support after congenital heart surgery. Data from a national registry have reported that cardiac patients have the poorest survival of all extracorporeal membrane oxygenation recipients. Concerns have been raised about the appropriateness of such an aggressive strategy, especially in light of the high costs and potential for long-term neurologic disability. We reviewed our experience with salvage cardiac extracorporeal membrane oxygenation to determine the cost-utility, which accounts for both costs and quality of life. Medical records of patients with congenital heart disease receiving salvage cardiac extracorporeal membrane oxygenation between January 2000 and May 2004 were reviewed. Charges for all medical care after the institution of extracorporeal membrane oxygenation were determined and converted to costs by published standards. The quality-of-life status of survivors was determined with the Health Utilities Index Mark II. Salvage cardiac extracorporeal membrane oxygenation was instituted in 32 patients (18 for cardiopulmonary arrest and 14 for cardiac failure after heart surgery) at a median age of 2.0 months (range, 4 days to 5.1 years). Congenital heart disease was present in 27 (84%). The mean duration of extracorporeal membrane oxygenation support was 5.1 +/- 4.1 days. Survival to hospital discharge was 50%, including 1 patient bridged to heart transplantation. Survival to 1 year was 47%. The mean score of the Health Utilities Index for the survivors was 0.75 +/- 0.19 (range, 0.41-1.0). The median cost for hospital stay after the institution of extracorporeal membrane oxygenation was USD 156,324 per patient. The calculated cost-utility for salvage extracorporeal membrane oxygenation in this population was USD 24,386 per quality-adjusted life-year saved, which would be considered within the range of accepted cost-efficacy (< USD 50,000 per quality-adjusted life-year saved). Salvage cardiac extracorporeal membrane oxygenation results in reasonable survival and is justified on a cost-utility basis.


Dynamics of extracellular polymeric substances in UASB and EGSB reactors treating medium and low concentrated wastewaters.
In this work the dynamic study of EPS (Extracellular Polymeric Substances) concentration and distribution during the operation of two different reactor configurations (UASB and EGSB) is presented, treating medium (5 g COD/l) and low-concentrated (0.5 g COD/l) wastewater. Medium-concentrated wastewater was supplied for granules maturation as well as for stabilisation of the process. The effect of substrate change on granule characteristics was followed in both reactors. Total concentration of EPS associated to steady operation, was higher in the UASB reactor. The change to a low-concentrated substrate led to an increased difference, promoting a sharp destabilisation of the EGSB reactor, observing an increment in filamentous structures, causing biomass flotation and wash out. Although total concentration of EPS remained almost constant in the UASB reactor, their composition and distribution presented significant differences. The ratio of protein/polysaccharides as well as acidic-polysaccharides/total (neutral + acidic) polysaccharides decreased drastically in the EGSB reactor, while in the UASB reactor, the decrease was not so important and not enough for destabilisation of granule structure. Moreover, polysaccharides distribution seemed to have an important role in granule stability being enough to maintain granule cohesion only in the case of the UASB reactor. These observations point to composition and distribution of EPS rather than their total concentration as key parameters for granule stability and settleability.


Stereoscopic digital subtraction angiography in neuroradiologic assessment.
Digital subtraction angiography (DSA) with stereoscopic imaging was performed in 40 patients for evaluation of a variety of cerebrospinal disorders. It was facilitated by a C-arm mounted x-ray tube and imaging chain with 7 degrees angulation between image pairs. Stereoscopic digital imaging proved particularly useful in the preoperative assessment of aneurysms, arteriovenous malformations, and primary and metastatic tumors. The technique was also found to be useful as a real-time adjunct to therapeutic radiographic procedures, as an aid in stereotaxic procedures, and in follow-up of postsurgical patients. Although the intravenous route was occasionally used, especially in postoperative follow-up of aneurysms, the procedure was most often carried out via an intraarterial approach. Stereoscopy was useful in supplying depth information regarding the relations between lesions and surrounding normal and abnormal vasculature. This technique combines the demonstrated advantages of intraarterial DSA with the unique advantage of stereoscopic imaging to demonstrate three-dimensional detail, thus contributing significantly to diagnostic confidence. Disadvantages are discussed. Further refinements in the equipment are expected: generation of stereo images with one injection, thus increasing procedure efficiency and patient safety; a video stereoscopic viewing unit; and the ability to obtain precise measurements via computer of depth, position, distance between, and true size of objects.


Evidence that the central region of glycoprotein IIIa participates in integrin receptor function.
We have obtained evidence that the ligand-recognition region of the integrin beta-subunit, platelet glycoprotein IIIa (GPIIIa), is discontinuous. Receptor function can be localized to residues near the N-terminus and to the central region of the polypeptide chain. The epitope recognized by our monoclonal antibody, CS-1, which substantially inhibits fibrin(ogen) binding to ADP- and thrombin-stimulated platelets [Ramsamooj, Doellgast & Hantgan (1990) Thromb. Res. 58, 577-592], is contained within residues 349-422 of GPIIIa. This sequence is adjacent to a proteinase-resistant domain of GPIIIa which is linked by disulphide bond(s) to an N-terminal segment near to the putative Arg-Gly-Asp recognition site [D'Souza, Ginsberg, Burke, Lam & Plow (1988) Science 242, 91-93]. Limited trypsin digestion of purified platelet GPIIIa yielded a mixture of two-chain molecules comprised of an N-terminal fragment disulphide-bonded to one of four fragments, which began at residues 299, 303, 353 or 423. Tryptic cleavage of the 300-422 segment correlated with loss of immunoreactivity with anti-GPIIIa monoclonal antibody, CS-1. Chymotrypsin cleavage of GPIIIa resulted in an N-terminal 19 kDa fragment joined by at least one intrachain cystine residue to a 46 kDa polypeptide beginning at residue 349. Partial reduction with dithiothreitol released the larger chymotryptic fragment with its epitope for CS-1 intact. These results have enabled us to localize the epitope recognized by our inhibitory monoclonal antibody, CS-1, to residues 349-422 of GPIIIa. Our data are consistent with a structure in which both the N-terminal and central regions of GPIIIa, which may be in close proximity in the functional GPIIb-IIIa complex, participate in ligand binding.


Crystallographic Characterization of the Carbonylated A-Cluster in Carbon Monoxide Dehydrogenase/Acetyl-CoA Synthase.
The Wood-Ljungdahl pathway allows for autotrophic bacterial growth on carbon dioxide, with the last step in acetyl-CoA synthesis catalyzed by the bifunctional enzyme carbon monoxide dehydrogenase/acetyl-CoA synthase (CODH/ACS). ACS uses a complex Ni-Fe-S metallocluster termed the A-cluster to assemble acetyl-CoA from carbon monoxide, a methyl moiety and coenzyme A. Here, we report the crystal structure of CODH/ACS from Moorella thermoacetica with substrate carbon monoxide bound at the A-cluster, a state previously uncharacterized by crystallography. Direct structural characterization of this state highlights the role of second sphere residues and conformational dynamics in acetyl-CoA assembly, the biological equivalent of the Monsanto process.


Recurrent lipoma-like hibernoma.
This is the fifth report of the rare lipoma-like variant of hibernoma, the 19th case reported, and the first documented as recurring after excision. Our patient was a 56-year-old man whose painful lipoma-like hibernoma (LLH) in the pelvis/buttock was initially treated with marginal excision 15 years earlier. Nine years after treatment, the LLH recurred locally. The recurrence was treated with partial excision and embolization, which alleviated symptoms. The disease was stable 26 months after repeat excision and 202 months after initial treatment. LLH is 1 of the 4 histologic variants of hibernomas, which are rare benign lipomatous tumors distinguished from other lipomas by their brown-fat component. Only minimal information is available regarding the clinicopathologic characteristics of the individual variants. Reviewing the literature, we found that LLH predominantly develops in the fifth and sixth decades, and mean age at diagnosis is 41 years (age range, 2-66 years). LLH has a slight male predilection of 10:9. It most commonly develops in the thigh, with other occurrences reported in the hip, trunk, knee, and calf. The magnetic resonance imaging signal of LLH mirrors fat in all sequences. Presence of internal septations and enhancement with contrast are variable. Histologically, LLH is defined as a hibernoma composed predominantly of univacuolated white-fat cells and only scattered granular or pale hibernoma cells. The literature provides only a few treatment details regarding this variant.


Relative Importance Assigned to Health Care Rationing Principles at the Bedside: Evidence From a Portuguese and Bulgarian Survey.
Activity was undertaken to develop a Prioritization Scoring Index for Portugal and Bulgaria that weights the importance given to ethical rationing principles that should guide decisions at the bedside. Data from two random samples of 355 Portuguese and 298 Bulgarian members of the public were collected from an online questionnaire. Questions asked about the level of importance given to specific issues related to patient's prioritization criteria. Responses were analyzed quantitatively with the SPSS. In the process of selecting the patient to treat, Portuguese and Bulgarian respondents seem unanimous in giving greater importance to (i) the treatment outcomes, (ii) the severity of illness, (iii) children, and (iv) patients' fragility. In general, Portuguese and Bulgarian respondents allocate more than 50% of the prioritization weight to equity considerations, approximately 35% to efficiency considerations, and 5% to lottery selection. Even so, Bulgarian respondents rate highly the equity and less the efficiency consideration than Portuguese respondents. Although the pursuit of efficiency seems to be valued by respondents, their major concern seems to be with the reduction of inequalities in health.


The coexpression of EphB4 and EphrinB2 is associated with poor prognosis in HER2-positive breast cancer.
HER2 overexpression is associated with aggressive phenotypes in breast cancer, including increased tumor proliferation, greater invasiveness, and reduced overall survival. The overall response rate to HER2-targeted therapies remains <30%. There is an urgent need for the identification of efficient markers to predict patients with a poor prognosis. This study was designed to investigate the correlation between EphB4 and EphrinB2 expression and the clinicopathological characteristics of HER2-positive breast cancer. A total of 111 primary HER2-positive breast cancer patients were enrolled in this study (diagnosed since December 2005 to November 2010 from the Second Hospital of Dalian Medical University). The protein expression of EphB4 and EphrinB2 was examined by immunohistochemistry using paraffin-embedded tumor tissues. There was a significant correlation between EphB4 and EphrinB2 expression (P=0.013, r=0.255). Kaplan-Meier analysis showed that the prognosis of patients with a high expression of both EphB4 and EphrinB2 was significantly worse than the prognosis of patients with either EphB4 or EphrinB2 expression and patients with negative expression (hazard ratio [HR] =1.935, P=0.0224). However, high expression of EphB4 or EphrinB2 alone was not an independent prognostic factor to predict worse overall survival. To summarize, HER2-positive breast cancer patients with overexpression of both EphB4 and EphrinB2 were associated with the worst prognosis. High expression of EphB4 and EphrinB2 correlated with poor overall survival, which can serve as an independent prognostic indicator in primary HER2-positive breast cancer patients.


Nonmalignant mortality among workers in six Norwegian aluminum plants.
This study investigated the associations between exposure to fluorides and polycyclic aromatic hydrocarbons (PAH) and mortality from nonmalignant diseases among workers in the Norwegian primary aluminum industry. Mortality among 10,857 men, employed for more than 3 years in 1 of 6 aluminum plants, was investigated from 1962 to 1996, giving 239,246 person-years during follow-up. Ajob-exposure matrix covering all 6 plants was used to estimate the individual exposure to total fluorides and particulate PAH. The observed cause-specific deaths were compared with expected figures calculated from national rates. Dose-response relations were investigated by internal comparisons using Poisson regression and by stratified analyses for standardized mortality ratio. Potential confounding by smoking was investigated in subanalyses restricted to 3 of the plants for which information on smoking habits was accessible. Mortality from circulatory disease was slightly lower than expected [SMR 0.95, 95% confidence interval (95%CI) 0.9-1.0], while there was an increased mortality from asthma, emphysema, and chronic bronchitis combined, SMR 1.2 (95% CI 1.0-1.5). Mortality from these diseases was associated with cumulative exposure to fluorides. The rate ratio in the internal analysis rose steadily to 2.5 (95% CI 1.5-4.3) for the upper exposure category. No association was observed between cumulative fluoride exposure or PAH exposure and circulatory mortality. The study showed an association between exposure to potroom emissions measured by fluorides and mortality from asthma, emphysema, and chronic bronchitis combined.


Genomic alterations and allelic imbalances are strong prognostic predictors in osteosarcoma.
Osteosarcoma, the most common primary malignant tumor of the bone, is characterized by complex karyotypes with numerous structural and numerical alterations. Despite attempts to establish molecular prognostic markers at the time of diagnosis, the most accepted predictive factor remains the histologic evaluation of necrosis after neoadjuvant chemotherapy. The present approach was carried out to search for genome-wide recurrent loss of heterozygosity and copy number variations that could have prognostic and therapeutic impact for osteosarcoma patients. Pretherapeutic biopsy samples of 45 osteosarcoma patients were analyzed using Affymetrix 10K2 high-density single nucleotide polymorphism arrays. Numerical aberrations and allelic imbalances were correlated with the histologically assessed response to therapy and clinical follow-up. The most frequent genomic alterations included amplifications of chromosome 6p21 (15.6%), 8q24 (15.6%, harboring MYC), and 12q14 (11.1%, harboring CDK4), as well as loss of heterozygosity of 10q21.1 (44.4%). All these aberrations and the total degree of heterozygosity of each tumor were significantly associated with an adverse outcome of patients and were used to define a chromosomal alteration staging system with a superior predictive potential compared with the histologic regression grading. Structural chromosomal alterations detected by single nucleotide polymorphism analysis provide a simple but robust parameter to anticipate response to chemotherapy. The proposed chromosomal alteration staging system might therefore help to better predict the clinical course of osteosarcoma patients at the time of initial diagnosis and to adapt neoadjuvant treatment in patients resistant to the current protocols.


Hydroxytyrosol Induces Dyslipidemia in an ApoB100 Humanized Mouse Model.
Extra virgin olive oil has numerous cardiopreventive effects, largely due to its high content of (poly)phenols such as hydroxytyrosol (HT). However, some animal studies suggest that its excessive consumption may alter systemic lipoprotein metabolism. Because human lipoprotein metabolism differs from that of rodents, this study examines the effects of HT in a humanized mouse model that approximates human lipoprotein metabolism. Mice are treated as follows: control diet or diet enriched with HT. Serum lipids and lipoproteins are determined after 4 and 8 weeks. We also analyzed the regulation of various genes and miRNA by HT, using microarrays and bioinformatic analysis. An increase in body weight is found after supplementation with HT, although food intake was similar in both groups. In addition, HT induced the accumulation of triacylglycerols but not cholesterol in different tissues. Systemic dyslipidemia after HT supplementation and impaired glucose metabolism are observed. Finally, HT modulates the expression of genes related to lipid metabolism, such as Pltp or Lpl. HT supplementation induces systemic dyslipidemia and impaired glucose metabolism in humanized mice. Although the numerous health-promoting effects of HT far outweigh these potential adverse effects, further carefully conducted studies are needed.


The minimal promoter of the human alpha 3 nicotinic receptor subunit gene. Molecular and functional characterization.
The minimal promoter of the human alpha(3) nicotinic receptor subunit gene has been mapped to a region of 60 base pairs and found to contain two Sp1 sites, one of which is essential for promoter activity. DNase footprinting has revealed the presence of another region of interaction with nuclear factors (named F2) immediately downstream of the Sp1 sites. This region has been found to be functional since it is capable of stimulating the minimal promoter. The F2 protection is completely and specifically competed by an AP2 consensus oligonucleotide that has been proved to bind AP2alpha exclusively. However, the AP2alpha recombinant protein was unable to bind the F2 region directly, thus suggesting that AP2alpha may participate in F2 protection by protein-protein interactions with other nuclear factors. The minimal promoter has been shown to be stimulated by two additional regions, one located downstream of F2 and the other upstream of the minimal promoter itself. In neuronal cells, the combined stimulatory activities of these three regions have synergistic effects, whereas in non-neuronal cells, there is a negative interference between the upstream and downstream regions. These opposite transcriptional effects may account for at least part of the neuro-specific expression profile of the alpha(3) gene.


Vortex wakes generated by robins Erithacus rubecula during free flight in a wind tunnel.
The wakes of two individual robins were measured in digital particle image velocimetry (DPIV) experiments conducted in the Lund wind tunnel. Wake measurements were compared with each other, and with previous studies in the same facility. There was no significant individual variation in any of the measured quantities. Qualitatively, the wake structure and its gradual variation with flight speed were exactly as previously measured for the thrush nightingale. A procedure that accounts for the disparate sources of circulation spread over the complex wake structure nevertheless can account for the vertical momentum flux required to support the weight, and an example calculation is given for estimating drag from the components of horizontal momentum flux (whose net value is zero). The measured circulations of the largest structures in the wake can be predicted quite well by simple models, and expressions are given to predict these and other measurable quantities in future bird flight experiments.


Tumor necrosis factor enhances SN38-mediated apoptosis in mesothelioma cells: the role of nuclear factor-kappaB pathway activation.
Despite the best and most aggressive, often integrated, standard therapeutic approaches for mesothelioma, overall survival remains very poor. The actual failure points out clearly the need for the development of novel therapy. One of the promising paths of experimentation is artificial induction of apoptosis. A therapeutic strategy that relies on the down-regulation of BCL-XL inhibition nuclear factor kappaB (NF-kappaB) with a combination of SN38 and tumor necrosis factor (TNF) was studied in human mesothelioma cell lines (MSTO-221H, IST-MES1, IST-MES2, MPP89, H28, H513, H2052, and H290). Cell proliferation (clonogenic assay) was inhibited strongly by the combination of TNF and SN38. Examining the persistence of the NF-kappaB complexes using an electrophoretic mobility-shift assay, it appeared that they still were present at 24 hours in TNF-treated cells. In SN38-treated cells, NF-kappaB complexes persisted for 6 hours. In cells that were treated with combined SN38 and TNF, NF-kappaB complexes disappeared quickly and became undetectable at 6 hours. In flow cytometry analysis, only cells that were treated with combined SN38 and TNF demonstrated significant cellular accumulation in the sub-G0-G1 phase, suggesting a specific induction of apoptosis. Morphologic examination (4,6-diamidino-2-phenylindole staining and electron microscopy) and internucleosomal DNA fragmentation (gel ladder) confirmed rigorously the induction of apoptosis. Because of NF-kappaB inhibition with the combination of SN38 and TNF, the expression of BCL-XL (both the protein [Western blot analysis] and the mRNA [reverse transcriptase-polymerase chain reaction analysis]) was down-regulated, cytochrome c was released into the cytoplasm, caspase 3 was activated (Western blot analysis), and, consequently, apoptosis was triggered. The authors hope that the results of the current study may contribute to the design and implementation of a novel therapeutic approach that improves patients' responses to treatment for mesothelioma.


Topiramate and vitamin e modulate antioxidant enzyme activities, nitric oxide and lipid peroxidation levels in pentylenetetrazol-induced nephrotoxicity in rats.
Previous studies have shown that generation of free radicals is increased following pentylenetetrazol kindling, due to increased cytosolic Ca2+ concentrations. Topiramate, a voltage-gated calcium channel inhibitor, has an evident effect in the treatment of childhood epilepsy; however, topiramate may cause nephrotoxicity. We investigated the effects of topiramate and vitamin E administration on pentylenetetrazol-induced nephrotoxicity in rats by evaluation of lipid peroxidation, nitric oxide, glutathione peroxidase, catalase and superoxide dismutase values. Forty male Wistar rats were randomly divided into five equal groups. Group 1 was used as control and group II received a single dose of pentylenetetrazol. Fifty and 100 mg/kg topiramate daily were intragastrically administered to rats in groups III and IV for 7 days, respectively. Intragastric 100 mg topiramate (daily for 7 days) and intraperitoneal vitamin E (150 mg/kg, daily for 3 days) combination were given to animals in group V before a single-dose pentylenetetrazol administration. Serum and kidney samples were taken after 3 hr of pentylenetetrazol administration. Pentylenetetrazol resulted in a significant increase in nitric oxide levels of serum and kidney, and lipid peroxidation levels of kidney although superoxide dismutase and catalase activities in the kidney was reduced by pentylenetetrazol administration. The lipid peroxidation levels in serum and kidneys and the nitric oxide levels in kidneys of groups III, IV and V were decreased by topiramate although the superoxide dismutase and catalase activities in the kidneys were increased. Lipid peroxidation and nitric oxide levels were reduced by the topiramate and vitamin E combination compared to only topiramate. Glutathione peroxidase activity was not affect by pentylenetetrazol, topiramate and vitamin E administrations. In conclusion, topiramate and vitamin E have protective effects on pentylenetetrazol-induced nephrotoxicity by inhibition of free radicals and by support of the antioxidant redox system.


Antibiotic prescriptions among dentists across Norway and the impact of COVID-19 pandemic.
The prescription of antibiotics in dental practice contributes significantly to the total use of antibiotics in primary healthcare. This study aimed to evaluate antibiotic prescription in dental practice during the years 2016-2021 in Norway and their relative contribution to national outpatient consumption and to investigate the influence of age, gender, geographic region, and COVID-19. A further aim was to review differences in prescribing patterns to verify effect of governmental strategies to reduce over-prescribing of antibiotics. This register study investigated the national antibiotic prescription between 2016 and 2021. Data was obtained from the Norwegian prescription register, the Norwegian Institute of Public Health and Statistics Norway. The consumption of 12 common antibiotics was measured using WHO defined daily doses (DDDs), DDD per 1000 inhabitants per day (DIDs 1000). A total of 6,049,445 antibiotic prescriptions of the 12 investigated compounds were issued in primary care during the study period. Dentists accounted for 942,350 prescriptions corresponding to 15.6% of the total. An overall decrease in the number of prescriptions by health professions other than dentists during the 5 years (IRR = 0.92, 95% CI:0.92-0.93, p < 0.001) was observed. For dentists a slight increase in the number of prescriptions (IRR = 1.01, 95% CI: 1.01-1.01, p < 0.001) was seen over the study period. The increase of antibiotic prescriptions in dentistry was more pronounced during the COVID-19 pandemic. The 4 most prescribed type of antibiotics based on average number of DDDs of the total period 2016-2021 were in descending order; phenoxymethylpenicillin (1,109,150) followed by amoxicillin (126,244), clindamycin (72,565), and metronidazole (64,599). An unexpected finding was that the prescription of the combination compound amoxicillin/clavulanic acid had significantly increased in dentistry during the last 5 years. Geographic, gender, and age differences in the rates of prescriptions were also seen. The data revealed that there are seasonal variations in dental prescriptions. Noticeable differences exist in prescribing patterns of antibiotics in the last 5 years. Restricted access to dental care due to COVID-19 may have resulted in increased antibiotic prescribing in dentistry as opposed to an otherwise downward trend. Despite national guidelines there is still a need for improvement of antibiotic stewardship in dentistry and to define effective methods to disseminate information.


Clinical, ultrasound and biochemical features of polycystic ovary syndrome in adolescents: implications for diagnosis.
Diagnosing polycystic ovary syndrome (PCOS) in adolescence is clinically challenging. The prevalence of clinical, ultrasound and biochemical features of PCOS in a community-based adolescent population using current diagnostic criteria has not previously been described. This was a prospective cohort study with 244 unselected post-menarchal girls, mean age 15.2 years, of whom 91% were Caucasian. Subjects were recruited from a large population-based birth cohort (the Raine cohort). Clinical hyperandrogenism (HA) was quantified using Ferriman-Gallwey scores. In the early follicular phase (Day 2-6), we measured circulating androgens and sex hormone-binding globulin by immunoassay, and ovarian morphology was assessed by transabdominal ultrasound examination. BMI and waist-hip ratio were measured. Normal ranges for early follicular phase androgens in adolescence were derived for this population. The top 5 and 10% of circulating free testosterone levels were 45.6 and 34.5 pmol/l, respectively. Fifty-one percent of girls reported menstrual irregularity. Clinical HA was uncommon, being observed in only 3.5% of girls. Mean ovarian volume was greater than that reported by others in adult women and 35% of girls had polycystic ovary morphology on transabdominal ultrasound. Taking the upper 5% of free testosterone as HA, 42 girls (18.5%) would have met the Rotterdam criteria for PCOS, 11 girls (5%) the Androgen Excess Society criteria and 7 girls (3.1%) the National Institutes of Health criteria. Menstrual irregularity is common in adolescence and does not relate to clinical or biochemical HA. Diagnostic criteria for PCOS which include ovarian volume and morphology may be of limited use in adolescence.


Increased Arm Swing and Rocky Surfaces Reduces Postural Control in Healthy Young Adults.
Fall-induced injuries can stem from a disruption in the postural control system and place a financial burden on the healthcare system. Most gait research focused on lower extremities and neglected the contribution of arm swing, which have been shown to affect the movement of the center of mass when walking. This study evaluated the effect of arm swing on postural control and stability during regular and rocky surface walking. Fifteen healthy young adults (age = 23.4 ± 2.8) walked on these two surfaces with three arm motions (normal, held, and active) using the CAREN Extended-System (Motek Medical, Amsterdam, NL). Mean, standard deviation and maximal values of trunk linear and angular velocity were calculated in all three axes. Moreover, step length, time and width mean and coefficient of variation as well as margin of stability mean and standard deviation were calculated. Active arm swing increased trunk linear and angular velocity variability and peak values compared to normal and held arm conditions. Active arm swing also increased participants' step length and step time, as well as the variability of margin of stability. Similarly, rocky surface walking increased trunk kinematics variability and peak values compared to regular surface walking. Furthermore, rocky surface increased the average step width while reducing the average step time. Though this surface type increased the coefficient of variation of all spatiotemporal parameters, rocky surface also led to increased margin of stability mean and variation. The spatiotemporal adaptations showed the use of "cautious" gait to mitigate the destabilizing effects of both the active arm swing and rocky surface walking and, ultimately, maintain dynamic stability.


Active acoustic telemetry tracking and tri-axial accelerometers reveal fine-scale movement strategies of a non-obligate ram ventilator.
California horn sharks (Heterodontus francisci) are nocturnally active, non-obligate ram ventilating sharks in rocky reef habitats that play an important ecological role in regulating invertebrate communities. We predicted horn sharks would use an area restricted search (ARS) movement strategy to locate dense resource patches while minimizing energetic costs of travel and nighttime activity. As ectotherms, we predicted environmental temperature would play a significant role in driving movement and activity patterns. Continuous active acoustic tracking methods and acceleration data loggers were used to quantify the diel fine-scale spatial movements and activity patterns of horn sharks. First passage time was used to identify the scale and locations of patches indicative of ARS. Activity was assessed using overall dynamic body acceleration (ODBA) as a proxy for energy expenditure. Behavior within a patch was characterized into three activity patterns: resting, episodic burst activity, and moderate, consistent activity. After resting in daytime shelters, individuals travelled to multiple reefs throughout the night, traversing through depths of 2-112 m and temperatures of 10.0-23.8 °C. All sharks exhibited area restricted search patch use and arrived at their first patch approximately 3.4 ± 2.2 h (mean ± SD) after sunset. Sharks exhibited moderate, consistent activity in 54% of the patches used, episodic burst activity in 33%, and few (13%) were identified as resting at night. ODBA peaked while sharks were swimming through relatively deeper (~ 30 m), colder channels when traversing from one patch to the next. There was no consistent pattern between ODBA and temperature. We provide one of the largest fine-scale, high-resolution paired data sets for an elasmobranch movement ecology study. Horn sharks exhibited ARS movement patterns for various activity patterns. Individuals likely travel to reefs known to have profitable and predictable patches, potentially tolerating less suitable environmental temperatures. We demonstrate how gathering high-resolution information on the movement decisions of a community resident enhances knowledge of community structure and overall ecosystem function.


Cytoplasmic components of acetylcholine receptor clusters of cultured rat myotubes: the 58-kD protein.
A 58-kD protein, identified in extracts of postsynaptic membrane from Torpedo electric organ, is enriched at sites where acetylcholine receptors (AChR) are concentrated in vertebrate muscle (Froehner, S. C., A. A. Murnane, M. Tobler, H. B. Peng, and R. Sealock. 1987. J. Cell Biol. 104:1633-1646). We have studied the 58-kD protein in AChR clusters isolated from cultured rat myotubes. Using immunofluorescence microscopy we show that the 58-kD protein is highly enriched at AChR clusters, but is also present in regions of the myotube membrane lacking AChR. Within clusters, the 58-kD protein codistributes with AChR, and is absent from adjacent membrane domains involved in myotube-substrate contact. Semiquantitative fluorescence measurements suggest that molecules of the 58-kD protein and AChR are present in approximately equal numbers. Differential extraction of peripheral membrane proteins from isolated AChR clusters suggests that the 58-kD protein is more tightly bound to cluster membrane than is actin or spectrin, but less tightly bound than the receptor-associated 43-kD protein. When AChR clusters are disrupted either in intact cells or after isolation, the 58-kD protein still codistributes with AChR. Clusters visualized by electron microscopy after immunogold labeling and quick-freeze, deep-etch replication show that, within AChR clusters, the 58-kD protein is sharply confined to AChR-rich domains, where it is present in a network of filaments lying on the cytoplasmic surface of the membrane. Additional actin filaments overlie, and are attached to, this network. Our results suggest that within AChR domains of clusters, the 58-kD protein lies between AChR and the receptor-associated 43-kD protein, and the membrane-skeletal proteins, beta-spectrin, and actin.


Characterisation of detergent-insoluble membranes in pollen tubes of Nicotiana tabacum (L.).
Pollen tubes are the vehicle for sperm cell delivery to the embryo sac during fertilisation of Angiosperms. They provide an intriguing model for unravelling mechanisms of growing to extremes. The asymmetric distribution of lipids and proteins in the pollen tube plasma membrane modulates ion fluxes and actin dynamics and is maintained by a delicate equilibrium between exocytosis and endocytosis. The structural constraints regulating polarised secretion and asymmetric protein distribution on the plasma membrane are mostly unknown. To address this problem, we investigated whether ordered membrane microdomains, namely membrane rafts, might contribute to sperm cell delivery. Detergent insoluble membranes, rich in sterols and sphingolipids, were isolated from tobacco pollen tubes. MALDI TOF/MS analysis revealed that actin, prohibitins and proteins involved in methylation reactions and in phosphoinositide pattern regulation are specifically present in pollen tube detergent insoluble membranes. Tubulins, voltage-dependent anion channels and proteins involved in membrane trafficking and signalling were also present. This paper reports the first evidence of membrane rafts in Angiosperm pollen tubes, opening new perspectives on the coordination of signal transduction, cytoskeleton dynamics and polarised secretion.


Biogenic amines in wines from three Spanish regions.
One hundred and sixty-three wines from La Rioja, Utiel-Requena, and Tarragona were analyzed to determine if there were any differences in the concentrations of six biogenic amines that are found in these three regions. The influence of grape variety, type of vinification, wine pH, malolactic fermentation, and storage in bottle on biogenic amine concentrations was studied. Results show important differences in putrescine and histamine concentrations among regions, varieties of grape, and type of wine; differences were less appreciable for the remaining biogenic amines studied. Low pH prevented biogenic amine formation. Malolactic fermentation and short storage periods in bottle (3-6 months) showed increases in histamine concentration, whereas longer periods of storage led to a general decrease in histamine. Several strains of lactic acid bacteria were isolated in this work, and their ability to form biogenic amines was assayed in synthetic media, grape must, and wine. Grape varieties, different types of winemaking, pH, and lactic acid bacteria may be responsible for the differences observed in the biogenic amine concentrations of the wines analyzed.


Establishment of a standardized system to perform population structure analyses with limited sample size or with different sets of SNP genotypes.
Recent studies have demonstrated that principal component analysis (PCA) can detect the presence of population mixture and admixture in a sample and thus can be used to correct population stratification in genome-wide association studies (GWAS). We propose a complementary approach to PCA that compensates for potential weaknesses associated with PCA, so that one can perform population structure analyses using limited numbers of subjects and single-nucleotide polymorphisms (SNPs). Our method first requires a PCA of the largest reference sample from a population to standardize the system. Once the system is established, it can perform PCA for each individual with a much smaller number of SNPs drawn from the same population. This is because of the introduction of the probabilistic PCA, so that the prediction of the principal components (PCs) is performed under a rigorous probabilistic framework. The subsequent linear discriminant analysis also helps to understand from which ancestries or subpopulations a given individual is more likely to derive, in terms of posterior probabilities given the predicted PCs. A real-world prototype of the system for the Japanese population is developed based on 19 260 subjects, which illustrates the potential usefulness of the system as an aid in the detection of population structures in validation samples, or to help with the correction of population stratification in GWAS.