Improved multiplex PCR using conserved and species-specific 16S rRNA gene primers for simultaneous detection of Actinobacillus actinomycetemcomitans, Bacteroides forsythus, and Porphyromonas gingivalis.
Among putative periodontal pathogens, Actinobacillus actinomycetemcomitans, Bacteroides forsythus, and Porphyromonas gingivalis are most convincingly implicated as etiological agents in periodontitis. Therefore, techniques for detection of those three species would be of value. We previously published a description of a multiplex PCR that detects A. actinomycetemcomitans and P. gingivalis. The present paper presents an improvement on that technique, which now allows more sensitive detection of all three periodontal pathogens. Sensitivity was determined by testing serial dilutions of A. actinomycetemcomitans, B. forsythus, and P. gingivalis cells. Primer specificity was tested against (i) all gene sequences from the GenBank-EMBL database, (ii) six A. actinomycetemcomitans, one B. forsythus, and four P. gingivalis strains, (iii) eight different species of oral bacteria, and (iv) supra- and subgingival plaque samples from 20 healthy subjects and subgingival plaque samples from 10 patients with periodontitis. The multiplex PCR had a detection limit of 10 A. actinomycetemcomitans, 10 P. gingivalis, and 100 B. forsythus cells. Specificity was confirmed by the fact that (i) none of our forward primers were homologous to the 16S rRNA genes of other oral species, (ii) amplicons of predicted size were detected for all A. actinomycetemcomitans, B. forsythus, and P. gingivalis strains tested, and (iii) no amplicons were detected for the eight other bacterial species. A. actinomycetemcomitans, B. forsythus, and P. gingivalis were detected in 6 of 20, 1 of 20, and 11 of 20 of supragingival plaque samples, respectively, and 4 of 20, 7 of 20, and 13 of 20 of subgingival plaque samples, respectively, from periodontally healthy subjects. Among patients with periodontitis, the organisms were detected in 7 of 10, 10 of 10, and 7 of 10 samples, respectively. The simultaneous detection of three periodontal pathogens is an advantage of this technique over conventional PCR assays.


Hepatic and renal effects of low concentrations of methoxyflurane in exposed delivery ward personnel.
During five alternating three-week periods either methoxyflurane-nitrous oxide or nitrous oxide alone was used for obstetrical analgesia. Delivery ward personnel were followed by venous blood samples once a week. Analyses of blood urea nitrogen, serum uric acid, SGOT and SGPT showed significantly elevated levels three days after exposure to methoxyflurane. This study demonstrates the importance of the scavenging of anesthetic gases to reduce the exposure of personnel to inhalational agents used in delivery suites. Since definite alterations in the indices of both hepatic and renal functions were recognized in obstetrical personnel following exposure, a re-evaluation of the use of methoxyflurane for obstetrical analgesia is suggested.


Supporting the old but neglecting the young? The two faces of ageism.
Ageism is the most prevalent form of prejudice and is experienced by both older and younger people. Little is known about whether these experiences are interdependent or have common origins. We analyze data from 8,117 older (aged 70 and over) and 11,647 younger respondents (15-29 years) in representative samples from 29 countries in the European Social Survey. Using multilevel structural equation modeling, we test the hypothesis that older people are less likely, and younger people more likely, to suffer age discrimination if they live in a country with stronger structural support for older people. We also test the hypothesis that although stronger social norm against age discrimination reduce age discrimination suffered by older people it does not inhibit discrimination against younger people. These hypotheses are supported, and the results underline the neglected problem of ageism toward youth. Findings highlight that strategies for reducing age prejudice must address ageism as a multigenerational challenge, requiring attention to intergenerational cohesion and resource distribution between ages. (PsycInfo Database Record (c) 2020 APA, all rights reserved).


Putative Intravascular Myofibroma Mimicking a Vascular Malformation With Phleboliths.
Myofibroma is a rare, benign myofibroblastic tumor that commonly presents at birth or in early infancy, usually as a painless, slow-growing, solitary, nodular mass. We present a case of a 40-year-old woman with a painful, solitary, myofibroma on the right elbow. The unique features of this case include age and gender of the patient, site, pain on presentation, tumor morphology, and putative intravascular nature of the tumor.


Development of the Chitra tilting disc heart valve prosthesis.
The high prevalence of rheumatic valvular disease in the young population and the high cost of imports necessitated the development of an Indian valve. The development of a tilting disc prosthesis was successfully concluded in February 1995, when the third model completed its clinical trial. The tilting disc valve has an integrally machined cobalt alloy cage, an ultra high molecular weight polyethylene disc and a polyester suture ring. The choice of design was based on its superior hydrodynamics and the age distribution of patients, the majority of whom were below 30 years. The polymer-metal combination was selected for its extremely low wear rate and proven durability in the human body. The hydrodynamic performance was tested under steady and pulsatile flow conditions. The accelerated durability of nine test valves was evaluated at 800-840 cycles/min for over 350 million cycles each. Size 23 mm valves mere implanted in the mitral position of five sheep. In a clinical trial, 306 patients with isolated mitral or aortic valve replacements were followed up for a total of 371 patient years (mean 1.37 years and range 0-4 years). The hydrodynamic performance was comparable to that of proven clinical models. The accelerated testing indicated lifetimes in excess of 50 years and the animal trials showed the valve to be safe. In the clinical trial, there was no incidence of structural failure or paravalvular leak. The linearized rate of late thromboembolism was 6.2%/patient-year (pty), anticoagulant related hemorrhage 0.54%/pty and infective endocarditis 0.54%/pty. At two years, the total actuarial survival was 89.5%. The higher incidence of thromboembolism and the very low incidence of anticoagulant related hemorrhage illustrate the difficulty in the management of anticoagulant therapy in a developing country, while the low incidence of endocarditis reflects their greater resistance to infection. These data clearly showed the valve to be safe and comparable to other similar valves in clinical use.


AIDS and haemophilia.
The prevalence of overt AIDS in haemophilia A in the USA is about 3% treated patients and about 2% in UK. Nevertheless up to 90% of patients with haemophilia A treated with commercial concentrates of factor VIII prepared from the blood of paid donors are seropositive to the AIDS-associated virus LAV/HTLV III. Infection seems to have commenced in 1978 in USA and in 1980 in Europe. The accession rate to the symptomatic group is not known. Seropositivity is much lower in patients treated with blood products derived from voluntarily donated blood but such patients, even when seronegative, often show cellular immune defects. An additional immunosuppressive agent in factor VIII concentrates may be responsible for this and could predispose to LAV infection. AIDS, immune defects and seropositivity are less common in patients with haemophilia B even when treated with factor IX concentrate derived from paid donors. The reasons for this anomaly are not known. The diagnosis, social, family and counselling problems of infected patients are surveyed and the prospects for effective anti-viral or immunotherapy assessed. Treatment policies using heat-treated factor concentrates and the impact of donor screening are reviewed. It is concluded that the outlook for uninfected haemophiliacs with regard to AIDS is now good.


Antibody covalent immobilization on carbon nanotubes and assessment of antigen binding.
Controlling the covalent bonding of antibodies onto functionalized carbon nanotubes is a key step in the design and preparation of nanotube-based conjugates for targeting cancer cells. For this purpose, an anti-MUC1 antibody (Ab) is linked to both multi-walled (MWCNTs) and double-walled carbon nanotubes (DWCNTs) using different synthetic strategies. The presence of the Ab attached to the nanotubes is confirmed by gel electrophoresis and thermogravimetric analysis. Most importantly, molecular recognition of the antigen by surface plasmon resonance is able to determine similar Ab binding capacities for both Ab-DWCNTs and Ab-MWCNTs. These results are very relevant for the design of future receptor-targeting strategies using chemically functionalized carbon nanotubes.


Molecular biology of hematopoietic stem cells.
Human CD34+ hematopoietic stem and progenitor cells are capable of maintaining a life-long supply of the entire spectrum of blood cells dependent on systemic needs. Recent studies suggest that hematopoietic stem cells are, beyond their hematopoietic potential, able to differentiate into nonhematopoietic cell types, which could open novel avenues in the field of cellular therapy. Here, we concentrate on the molecular biology underlying basic features of hematopoietic stem cells. Immunofluorescence analyses, culture assays, and transplantation models permit an extensive immunological as well as functional characterization of human hematopoietic stem and progenitor cells. New methods such as cDNA array technology have demonstrated that distinct gene expression patterns of transcription factors and cell cycle genes molecularly control self-renewal, differentiation, and proliferation. Furthermore, several adhesion molecules have been shown to play an important role in the regulation of hematopoiesis and stem cell trafficking. Progress has also been made in elucidating molecular mechanisms of stem cell aging that limit replicative potential. Finally, more recent data provide the first molecular basis for a better understanding of transdifferentiation and developmental plasticity of hematopoietic stem cells. These findings could be helpful for non-hematopoietic cell therapeutic approaches.


Moth declines are most severe in broadleaf woodlands despite a net gain in habitat availability.
While agricultural intensification and habitat loss are cited as key drivers of moth decline, these alone cannot explain declines observed in UK woodlands - a habitat that has expanded in area since 1968.We quantified how moth communities changed across habitats and regions and determined how species traits interacted with habitat in predicting moth abundance change. We hypothesised that, in woodlands, species more vulnerable to shading and browsing by deer (species specialising on forbs, shrubs and shade-intolerant plants) had declined more severely than other species, and that moth decline in woodlands was more severe at sites more susceptible to deer damage.We modelled abundance, biomass, species richness and diversity from 1968 to 2016 and explored how these interacted with habitat and region. We also modelled the interaction between habitat and two moth species traits: larval feeding guild and shade-tolerance of hostplant.Moth declines were consistently highest in broadleaf woodland. Abundance, biomass, species richness and diversity declined significantly by -51%, -52%, -14% and -15% in woodlands, respectively, compared to national trends of -34%, -39%, -1% (non-significant) and +10%. Declines were no greater in woodlands more susceptible to deer browsing damage. Traits based analysis found no evidence that shading and intensive browsing by deer explained moth declines in woodland.Moth decline was more severe in broadleaf woodlands than in intensively managed farmlands. We found no evidence that deer browsing or increased shading has driven these trends: the primary cause of the decline of moths in woodlands remains unclear.


Socio-demographic characteristics of street children in rural communities undergoing urbanization.
The concept of the street child in rural communities has received little attention. This study describes the sociodemographic characteristics of the street children found in a group of rural communities. This descriptive study is nested in a cross sectional analytical study of street children in a group of rural communities undergoing urbanization. A cluster sample of street children as defined by the United Nations was taken in the seven chosen political wards. Majority of the street children (88.9%) were within the age group 15-17 years with more males (58.3%) than females (41.7%). Up to 64.7% had attained secondary level education while only 3.9% had no formal education. A high percentage, (61.4%) were still attending school and 15.8% had no work. Of those who work on the streets, being an apprentice (16.4%), petty trading (15.0%), part-time driving (9.5%) and car washing (5.0%) were the commonest types of work. Of those still schooling, 41.6% had no form of part-time work on the streets. None of the street children lived on the street with 65% still living with parents. However, 75% had been involved in the street life for 1-2 years with a median time of 2 years. More than 30% of their parents work outside town. The street child in rural communities differs from the urban perception which often has to do with those living rough and existing outside the family framework. More studies would be needed on the driving factors for street life in rural communities undergoing urbanization.


Multiplexing of injury codes for the parallel operation of enzyme logic gates.
The development of a highly parallel enzyme logic sensing concept employing a novel encoding scheme for the determination of multiple pathophysiological conditions is reported. The new concept multiplexes a contingent of enzyme-based logic gates to yield a distinct 'injury code' corresponding to a unique pathophysiological state as prescribed by a truth table. The new concept is illustrated using an array of NAND and AND gates to assess the biomedical significance of numerous biomarker inputs including creatine kinase, lactate dehydrogenase, norepinephrine, glutamate, alanine transaminase, lactate, glucose, glutathione disulfide, and glutathione reductase to assess soft-tissue injury, traumatic brain injury, liver injury, abdominal trauma, hemorrhagic shock, and oxidative stress. Under the optimal conditions, physiological and pathological levels of these biomarkers were detected through either optical or electrochemical techniques by monitoring the level of the outputs generated by each of the six logic gates. By establishing a pathologically meaningful threshold for each logic gate, the absorbance and amperometric assays tendered the diagnosis in a digitally encoded 6-bit word, defined as an 'injury code'. This binary 'injury code' enabled the effective discrimination of 64 unique pathological conditions to offer a comprehensive high-fidelity diagnosis of multiple injury conditions. Such processing of relevant biomarker inputs and the subsequent multiplexing of the logic gate outputs to yield a comprehensive 'injury code' offer significant potential for the rapid and reliable assessment of varied and complex forms of injury in circumstances where access to a clinical laboratory is not viable. While the new concept of parallel and multiplexed enzyme logic gates is illustrated here in connection to multi-injury diagnosis, it could be readily extended to a wide range of practical medical, industrial, security and environmental applications.


New plastoquinones isolated from the brown alga, Sargassum micracanthum.
Four new plastoquinones were isolated from the methanolic extract of the brown alga, Sargassum micracanthum. Their structures were elucidated based on spectroscopic analysis and chemical conversions from 2-geranylgeranyl-6-methyl-1,4-benzohydroquinone. These plastoquinones exhibited significant antioxidant activities such as an inhibitory effect on lipid peroxidation and a radical scavenging effect on 1,1-diphenyl-2-picrylhydrazyl (DPPH). Some of the new plastoquiones showed cytotoxic activity against cancer cell line.


Diverticular disease: threee studies. Part II - Treatment with bran.
Forty patients with diverticular disease were treated with wheat bran 24g/day for at least six months. Thirty-three patients showed a very satisfactory clinical response. Sixty per cent of all symptoms were abolished, and a further 28% were relieved. After treatment the transit times accelerated in patients whose initial times were slower than 60 hours and slowed down in those whose initial transit times were faster than 36 hours. Stool weight increased significantly. The number of intracolonic high pressure waves decreased, especially during and after eating. Barium enema studies showed less spasm in eight patients and no diverticula in three patients after taking bran. As well as relieving the symptoms of diverticular disease a high-fibre diet may also prevent the condition from developing.


The association between church attendance and psychological health in Northern Ireland: a national representative survey among adults allowing for sex differences and denominational difference.
This study extends previous research concerning the association between religion and psychological health in six ways: (1) by focusing clearly on religious attendance (church attendance); (2) by employing a robust measure of psychological distress (GHQ-12); (3) by studying a highly religious culture (Northern Ireland); (4) by taking sex differences into account (male or female); (5) by taking denominational differences into account (Catholic or Protestant); (6) and by obtaining a national representative sample (N = 4,281 adults aged 16 and above). Results from a 2 (sex) by 2 (denomination) ANCOVA demonstrated that Catholics recorded significantly lower levels of psychological health compared to Protestants, and that females showed significantly lower levels of psychological health compared to males. In addition, females reported higher frequency of religious service attendance than males, and Catholics reported higher attendance rates than Protestants. A significant positive association was found between frequency of religious attendance and GHQ-12 scores, and this association was moderated by sex and denomination. In conclusion, the results suggest that there may be sex and denominational differences in further understanding the relationship between frequency of religious attendance and psychological health.


Panhandle and reverse-panhandle PCR enable cloning of der(11) and der(other) genomic breakpoint junctions of MLL translocations and identify complex translocation of MLL, AF-4, and CDK6.
We used panhandle PCR to clone the der(11) genomic breakpoint junction in three leukemias with t(4;11) and devised reverse-panhandle PCR to clone the breakpoint junction of the other derivative chromosome. This work contributes two elements to knowledge on MLL translocations. First is reverse-panhandle PCR for cloning breakpoint junctions of the other derivative chromosomes, sequences of which are germane to understanding the MLL translocation process. The technique revealed duplicated sequences in one case of infant acute lymphoblastic leukemia (ALL) and small deletions in a case of treatment-related ALL. The second element is discovery of a three-way rearrangement of MLL, AF-4, and CDK6 in another case of infant ALL. Cytogenetic analysis was unsuccessful at diagnosis, but suggested t(4;11) and del(7)(q21q31) at relapse. Panhandle PCR analysis of the diagnostic marrow identified a breakpoint junction of MLL intron 8 and AF-4 intron 3. Reverse-panhandle PCR identified a breakpoint junction of CDK6 from band 7q21-q22 and MLL intron 9. CDK6 encodes a critical cell cycle regulator and is the first gene of this type disrupted by MLL translocation. Cdk6 is overexpressed or disrupted by translocation in many cancers. The in-frame CDK6-MLL transcript is provocative with respect to a potential contribution of the predicted Cdk6-MLL fusion protein in the genesis of the ALL, which also contains an in-frame MLL-AF4 transcript. The sequences in these three cases show additional MLL genomic breakpoint heterogeneity. Each breakpoint junction suggests nonhomologous end joining and is consistent with DNA damage and repair. CDK6-MLL is a new fusion of both genes.


Calgary, Edmonton and the University of Alberta: the extraordinary medical mobilization by Canada's newest province.
The Canadian contribution of medical services to the British Empire during the First World War was a national endeavour. Physicians from across the country enlisted in local regiments to join. No other region provided more physicians per capita than the newly formed province of Alberta. Largely organized through the Medical School of the University of Alberta, the No. 11 Canadian Field Ambulance out of Edmonton and the No. 8 Canadian Field Ambulance out of Calgary ultimately enlisted between one-third and half of the province's doctors to the war campaign. Many individuals from this region distinguished themselves, including LCol J.N. Gunn from Calgary, who commanded the No. 8 Canadian Field Ambulance; Maj Heber Moshier, one of the founders of the School of Pharmacy at the University of Alberta; and Dr. A.C. Rankin, who would go on to be the first Dean of Medicine at the University of Alberta. These Canadian heroes, and the many others like them who served with the No. 8 and 11 Field Ambulances, personify the sacrifice, strength and resilience of the medical community in Alberta and should not be forgotten.


Dammarane glycosides from the root of Machilus yaoshansis.
Nine new dammarane triterpene glycosides (1-3 and 8-13) and 12 known analogues have been isolated from an ethanol extract of the roots of Machilus yaoshansis. Compounds 1-7 have an uncommon 20,23-dihydroxydammar-24-en-21-oic acid-21,23-lactone moiety that was previously reported in compounds isolated from Gynostemma pentaphyllum. The configurations of the lactone moieties in 1-3 were determined by comparison of the experimental ECD spectra of 1-3 and the hydrolysates, 1a and 1b, with the corresponding calculated ECD spectra. On the basis of NMR and ECD data analysis of 1-7, the previously reported C-20 and C-23 configurations of 4-7 and related derivatives from Gynostemma pentaphyllum were revised. In addition, the application of NMR data and Cotton effects to the determination of the relative and absolute configurations of the γ-lactone moiety in 3β,20,23-trihydroxydammar-24-en-21-oic acid-21,23-lactone derivatives is discussed.


Subunits of the heterotrimeric transcription factor NF-Y are imported into the nucleus by distinct pathways involving importin beta and importin 13.
The transcriptional activator NF-Y is a heterotrimeric complex composed of NF-YA, NF-YB, and NF-YC, which specifically binds the CCAAT consensus present in about 30% of eukaryotic promoters. All three subunits contain evolutionarily conserved core regions, which comprise a histone fold motif (HFM) in the case of NF-YB and NF-YC. Our results of in vitro binding studies and nuclear import assays reveal two different transport mechanisms for NF-Y subunits. While NF-YA is imported by an importin beta-mediated pathway, the NF-YB/NF-YC heterodimer is translocated into the nucleus in an importin 13-dependent manner. We define a nonclassical nuclear localization signal (ncNLS) in NF-YA, and mutational analysis indicates that positively charged amino acid residues in the ncNLS are required for nuclear targeting of NF-YA. Importin beta binding is restricted to the monomeric, uncomplexed NF-YA subunit. In contrast, the nuclear import of NF-YB and NF-YC requires dimer formation. Only the NF-YB/NF-YC dimer, but not the monomeric components, are recognized by importin 13 and are imported into the nucleus. Importin 13 competes with NF-YA for binding to the NF-YB/NF-YC dimer. Our data suggest that a distinct binding platform derived from the HFM of both subunits, NF-YB/NF-YC, mediates those interactions.


Synthesis and cell adhesive properties of linear and cyclic RGD functionalized polynorbornene thin films.
Described herein is the efficient synthesis and evaluation of bioactive arginine-glycine-aspartic acid (RGD) functionalized polynorbornene-based materials for cell adhesion and spreading. Polynorbornenes containing either linear or cyclic RGD peptides were synthesized by ring-opening metathesis polymerization (ROMP) using the well-defined ruthenium initiator [(H(2)IMes)(pyr)(2)(Cl)(2)Ru═CHPh]. The random copolymerization of three separate norbornene monomers allowed for the incorporation of water-soluble polyethylene glycol (PEG) moieties, RGD cell recognition motifs, and primary amines for postpolymerization cross-linking. Following polymer synthesis, thin-film hydrogels were formed by cross-linking with bis(sulfosuccinimidyl) suberate (BS(3)), and the ability of these materials to support human umbilical vein endothelial cell (HUVEC) adhesion and spreading was evaluated and quantified. When compared to control polymers containing either no peptide or a scrambled RDG peptide, polymers with linear or cyclic RGD at varying concentrations displayed excellent cell adhesive properties in both serum-supplemented and serum-free media. Polymers with cyclic RGD side chains maintained cell adhesion and exhibited comparable integrin binding at a 100-fold lower concentration than those carrying linear RGD peptides. The precise control of monomer incorporation enabled by ROMP allows for quantification of the impact of RGD structure and concentration on cell adhesion and spreading. The results presented here will serve to guide future efforts for the design of RGD functionalized materials with applications in surgery, tissue engineering, and regenerative medicine.


Dose assessment for daily cone-beam CT in lung radiotherapy patients and its combination with treatment planning.
With the increased use of X-ray imaging for patient alignment in external beam radiation therapy, particularly with cone-beam computed tomography (CBCT), the additional dose received by patients has become of greater consideration. In this study, we analysed the radiation dose from CBCT for clinical lung radiotherapy and assessed its relative contribution when combined with radiation treatment planning for a variety of lung radiotherapy techniques. The Monte Carlo simulation program ImpactMC was used to calculate the 3D dose delivered by a Varian TrueBeam linear accelerator to patients undergoing thorax CBCT imaging. The concomitant dose was calculated by simulating the daily CBCT irradiation of ten lung cancer patients. Each case was planned with a total dose of 50-60 Gy to the target lesion in 25-30 fractions using the 3DCRT or IMRT plan and retrospectively planned using VMAT. For each clinical case, the calculated CBCT dose was summed with the planned dose, and the dose to lungs, heart, and spinal cord were analysed according to conventional dose conformity metrics. Our results indicate greater variations in dose to the heart, lungs, and spinal cord based on planning technique, (3DCRT, IMRT, VMAT) than from the inclusion of daily cone-beam imaging doses over 25-30 fractions. The average doses from CBCT imaging per fraction to the lungs, heart and spinal cord were 0.52 ± 0.10, 0.49 ± 0.15 and 0.39 ± 0.08 cGy, respectively. Lung dose variations were related to the patient's size and body composition. Over a treatment course, this may result in an additional mean absorbed dose of 0.15-0.2 Gy. For lung V5, the imaging dose resulted in an average increase of ~ 0.6% of the total volume receiving 5 Gy. The increase in V20 was more dependent on the planning technique, with 3DCRT increasing by 0.11 ± 0.09% with imaging and IMRT and VMAT increasing by 0.17 ± 0.05% and 0.2 ± 0.06%, respectively. In this study, we assessed the concomitant dose for daily CBCT lung cancer patients undergoing radiotherapy. The additional radiation dose to the normal lungs from daily CBCT was found to range from 0.15 to 0.2 Gy when the patient was treated with 25-30 fractions. Consideration of potential variation in relative biological effectiveness between kilovoltage imaging and megavoltage treatment dose was outside the scope of this study. Regardless of this, our results show that the assessment of imaging dose can be incorporated into the treatment planning process and the relative effect on overall dose distribution was small compared to the difference among planning techniques.


Targeting the interleukin-4 and interleukin-13 pathways in severe asthma: current knowledge and future needs.
Severe asthma is a heterogeneous disease that can be classified into phenotypes and endotypes based upon clinical or biological characteristics. Interleukin (IL)-4 and IL-13 play a key role in type 2 (T2) asthma. This article reviews the signaling pathway of IL-4 and IL-13 and highlights its targeted therapy in severe asthma. Several clinical trials of biologics targeting the IL-4/IL-13 pathway have recently been completed. In patients with severe, uncontrolled asthma, targeting IL-13 alone with biologics including lebrikizumab and tralokinumab has not shown consistent reduction in asthma exacerbations. Simultaneous targeting of both IL-4 and IL-13 by blocking IL-4 receptor α using dupilumab has yielded more consistent results in reducing asthma exacerbations and improving lung function, especially in patients with increased blood eosinophils. Other biomarkers of T2 inflammation such as exhaled nitric oxide and serum periostin may also predict response to biologics targeting the IL-4/IL-13 pathway. No biologic targeting the IL-4/IL-13 pathway is currently available for treatment of asthma, but emerging data suggest that biologics targeting IL-4 and IL-13 together may benefit patients with T2 high asthma. Additional data are needed about long-term efficacy and safety prior to incorporating these drugs into routine clinical practice.


Effect of topical timolol on the human retinal circulation.
The effect of topical timolol maleate 0.5% on the retinal circulation was investigated in 14 normal subjects using laser Doppler velocimetry and monochromatic fundus photography. In a double masked, randomized design, one eye received two drops of timolol maleate 0.5% and the fellow eye received two drops of placebo. Vessel diameter, maximum velocity of red blood cells, and volumetric blood flow rate were determined in a major temporal vein of each eye just prior to the instillation of drops, and then 90 min later. In comparison to the baseline value, there was a significantly larger average percentage increase in maximum velocity of red blood cells and volumetric blood flow rate in the timolol-treated eyes (11.0% and 13.2%, respectively) than in the control eyes (2.2% and 1.5%, respectively, paired t-test, P less than 0.05). No significant change in venous diameter was detected. No significant linear correlations were found between the percentage change in maximum velocity of red blood cells and mean brachial artery blood pressure, and between the percentage change in volumetric blood flow rate and mean brachial artery pressure in the placebo-treated eyes, whereas significant correlations were present between these quantities in the timolol-treated eyes (P less than 0.01 and P less than 0.05, respectively). The lack of such correlations in the control eyes is probably due to autoregulation of the retinal circulation. The presence of correlations in the timolol-treated eyes suggests that the drug may affect the capacity of the retina to autoregulate.


Experimental and Theoretical Investigation on Phase Formation and Mechanical Properties in Cr-Co-Ni Alloys Processed Using a Novel Thin-Film Quenching Technique.
The Cr-Co-Ni system was studied by combining experimental and computational methods to investigate phase stability and mechanical properties. Thin-film materials libraries were prepared and quenched from high temperatures up to 700 °C using a novel quenching technique. It could be shown that a wide A1 solid solution region exists in the Cr-Co-Ni system. To validate the results obtained using thin-film materials libraries, bulk samples of selected compositions were prepared by arc melting, and the experimental data were additionally compared to results from DFT calculations. The computational results are in good agreement with the measured lattice parameters and elastic moduli. The lattice parameters increase with the addition of Co and Cr, with a more pronounced effect for the latter. The addition of ∼20 atom % Cr results in a similar hardening effect to that of the addition of ∼40 atom % Co.


Renal trapping of In-111 troponolate labeled platelets in thrombotic thrombocytopenic purpura.
A patient with active thrombotic thrombocytopenic purpura underwent examination with In-111 troponolate labeled autologous platelets. A mean platelet survival of 1.2 days was found, comparable to formerly described studies using other methods. Furthermore, remarkable renal uptake of radioactivity consistent with abnormal platelet trapping was demonstrated. In the absence of any other signs of renal involvement, the conclusion was reached that the platelet scan is a sensitive and direct method for showing renal involvement in thrombotic thrombocytopenic purpura.


Secular trends towards delayed onsets of pathologies and prolonged longevities in Japanese patients with Werner syndrome.
Recent cases of increasingly elderly Werner Syndrome (WS) patients have paralleled increased lifespan in the general population, however, historical temporal lifespan variations in WS have not yet been ascertained. To assess temporal changes in life-span and progeroid comorbidity in WS, all Japanese WS patients documented from 1966-2004 were analyzed for age at onset of diabetes mellitus (DM), malignancy, and death. Of 1,019 WS analyzed, average age significantly increased for all variables studied over the study period. Average age of onset of malignancy and DM in all WS increased from 35.8 to 48.8 years and from 34.9 to 39.7 years, respectively (p < 0.001), while age at death increased from 38.2 to 52.8 years (p < 0.001), vs. 71.7 to 82.7 years (p < 0.001) in the general population. Lifespan increases in WS and the general population suggest a common environmental influence. Unlike the general population, no gender-specific difference in life-span occurred in WS, suggesting a gender-specific differential environmental effect on mutated WRN. Identification of factors responsible for age differences could facilitate improvement in survival and ageing phenotypes of WS patients and the general population.


[Ten myths about informed consent].
The author reviews ten myths that, in his opinion, make up the perspective of many professionals on informed consent (IC). A viewpoint, on the other hand, that is imprecise and of low ethical and juridical quality. The myths are: 1. IC is something foreign to the moral duties of doctors. 2. IC consists in getting the patients to sign the form authorising an intervention. 3. Good clinical practice consists in carrying out clinical actions well, from a scientific-technical point of view; IC has nothing to do with this. 4. The ethical foundation of IC is the principle of autonomy. 5. Obtaining IC is an obligation of the professionals who carry out interventions, but not of those who prescribe them. 6. IC is meaningless because the majority of patients do not want to be informed. 7. Patients have the right to refuse a course of treatment, but only before it is applied to them. Once it is started, the professionals cannot withdraw it. 8. The evaluation of the capacity of the patients to make decisions is the responsibility of the psychiatrists. 9. Respect for the requirements of IC can be satisfied without the health organisations investing additional resources in this. 10. Obtaining IC is a responsibility of the doctors but not of the nursing staff. The conclusion insists on the importance of exposing the hidden discourse of these myths in order to be able to change the practices concerning IC in our health organisations.


Long-term clinical outcomes of catheter ablation in patients with atrial fibrillation predisposing to tachycardia-bradycardia syndrome: a long pause predicts implantation of a permanent pacemaker.
There is a controversy as to whether catheter ablation should be the first-line therapy for tachycardia-bradycardia syndrome (TBS) in patients with atrial fibrillation (AF). We aimed to investigate long-term clinical outcomes of catheter ablation in patients with TBS and AF. Among 145 consecutive patients who underwent catheter ablation of AF with TBS, 121 patients were studied. Among 121 patients, 11 (9.1%) received implantation of a permanent pacemaker during a mean 21 months after ablation. Length of pause on termination of AF was significantly greater in patients who received pacemaker implantation after ablation than those who underwent ablation only (7.9 ± 3.5 vs. 5.1 ± 2.1 s, p < 0.001). Using a multivariate model, a long pause of 6.3 s or longer after termination of AF was associated with the requirement to implant a permanent pacemaker after ablation (HR 1.332, 95% CI 1.115-1.591, p = 0.002). This study suggests that, in patients with AF predisposing to TBS, long pause on termination of AF predicts the need to implant a permanent pacemaker after catheter ablation.


Dietary patterns of antioxidant vitamin and carotenoid intake associated with bone mineral density: findings from post-menopausal Japanese female subjects.
Recent studies show that antioxidants may reduce the risk of osteoporosis. This study showed the associations of bone mineral density with dietary patterns of antioxidant vitamins and carotenoids. The findings suggest the combination of vitamin C and β-cryptoxanthin intakes might provide benefit to bone health in post-menopausal Japanese female subjects. Recent epidemiological studies show antioxidants may reduce the risk of osteoporosis, but little is known about the dietary patterns of antioxidant vitamin and carotenoid intakes and their relation with bone mineral density (BMD). A total of 293 post-menopausal female subjects who had received health examinations in the town of Mikkabi, Shizuoka Prefecture, Japan, participated in the study. Radial BMD was measured using dual-energy X-ray absorptiometry. Dietary intakes of antioxidant vitamins and carotenoids were assessed by using a validated food-frequency questionnaire. Dietary patterns were identified on a selected set of antioxidants through principal component factor analysis. Three dietary patterns were identified. The "retinol" pattern, characterized by notably high intakes of preformed retinol, zeaxanthin, and vitamin E, was positively associated with the risk for low BMD. In contrast, the "β-cryptoxanthin" pattern, characterized by notably high intakes of β-cryptoxanthin and vitamin C, was negatively associated with low BMD. The odds ratios for low BMD in the highest tertiles of dietary intakes of preformed retinol, vitamin C, and β-cryptoxanthin against the lowest tertiles were 3.22 [95% confidence interval (CI), 1.38-7.51], 0.25 (CI, 0.10-0.66), and 0.40 (CI, 0.17-0.92), respectively, after adjustments for confounders. However, negative associations of vitamin C and β-cryptoxanthin with low BMD were not significant after further adjustment for intake of β-cryptoxanthin or vitamin C, respectively. Higher intakes of both vitamin C and β-cryptoxanthin were significantly associated with low BMD (P < 0.05). The combination of vitamin C and β-cryptoxanthin may be associated with radial BMD in post-menopausal Japanese female subjects.


Latent profiles of children's autonomic nervous system reactivity early in life predict later externalizing problems.
Prior researchers have observed relations between children's autonomic nervous system reactivity and externalizing behavior problems, but rarely considers the role of developmentally regulated changes in children's stress response systems. Using growth mixture modeling, the present study derived profiles of parasympathetic nervous system reactivity (as indicated by respiratory sinus arrhythmia (RSA)) and sympathetic nervous system reactivity (as indicated by pre-ejection period (PEP)) from low income, primarily Mexican American children measured repeatedly from infancy through age 5 (N = 383) and investigated whether profiles were associated with externalizing problems at age 7. Analyses identified two profiles of RSA reactivity (reactive decreasing and U-shaped reactivity) and three profiles of PEP reactivity (blunted/anticipatory reactivity, reactive decreasing, non-reactive increasing). Compared to children with an RSA profile of reactive decreasing, those with an RSA profile of U-shaped reactivity had marginally higher externalizing problems, however, this difference was not statistically significant. Children who demonstrated a profile of blunted/anticipatory PEP reactivity had significantly higher externalizing problems compared to those with a profile of non-reactive increasing, likely related to the predominantly male composition of the former profile and predominantly female composition of the latter profile. Findings contribute to our understanding of developmental trajectories of ANS reactivity and highlight the utility of a longitudinal framework for understanding the effects of physiological risk factors on later behavior problems.


Does the level of prostate cancer risk affect cancer prevention with finasteride?
Finasteride reduced the risk of prostate cancer by 24.8% in the Prostate Cancer Prevention Trial (PCPT). Whether this represents treatment or prevention and who is most likely to benefit are unknown. We sought to clarify these issues by this investigation. We fit a logistic regression model to men in the placebo group of the PCPT using risk factors for prostate cancer at entry to predict prostate cancer during the subsequent 7 years of study. Men in the two treatment groups were categorized into quintiles of risk of prostate cancer based on the predictive logistic model. A second model was fit evaluating finasteride's effect on prostate cancer for each subgroup defined by quartiles of baseline prostate-specific antigen (PSA) . The magnitude of the prevention effect of finasteride on prostate cancer was then evaluated across risk and PSA strata. Finasteride significantly reduced prostate cancer risk for all risk quintiles. For quintiles 1 through 5, odds ratios were 0.72, 0.52, 0.64, 0.66, and 0.71, respectively (all P < or = 0.05). For quartiles of risk of entry PSA (less than 0.7 ng/mL, 0.7 to 1.1 ng/mL, 1.1 to 1.7 ng/mL, and 1.8 to 3.0 ng/mL), odds ratios increased (smaller treatment effect) as PSA increased: 0.60, 0.62, 0.66, and 0.69, respectively, but remained significant for all strata (each P < 0.001). Finasteride significantly reduced prostate cancer risk regardless of the level of this risk, estimated either by multivariable risk or by PSA stratum; this suggests that finasteride exerts both treatment and preventive effects. All men undergoing PSA screening should be informed of the potential for finasteride to reduce their risk of prostate cancer.


Decomposing socioeconomic inequality in dental caries in Iran: cross-sectional results from the PERSIAN cohort study.
The current study aimed to measure and decompose socioeconomic-related inequalities in DMFT (decayed, missing, and filled teeth) index among adults in Iran. The study data were extracted from the adult component of Prospective Epidemiological Research Studies in IrAN (PERSIAN) from 17 centers in 14 different provinces of Iran. DMFT score was used as a measure of dental caries among adults in Iran. The concentration curve and relative concentration index (RC) was used to quantify and decompose socioeconomic-related inequalities in DMFT. A total of 128,813 adults aged 35 and older were included in the study. The mean (Standard Deviation [SD]) score of D, M, F and DMFT of the adults was 3.3 (4.6), 12.6 (10.5), 2.1 (3.4) and 18.0 (9.5), respectively. The findings suggested that DMFT was mainly concentrated among the socioeconomically disadvantaged adults (RC = - 0.064; 95% confidence interval [CI), - 0.066 to - 0.063). Socioeconomic status, being male, older age and being a widow or divorced were identified as the main factors contributing to the concentration of DMFT among the worse-off adults. It is recommended to focus on the dental caries status of socioeconomically disadvantaged groups in order to reduce socioeconomic-related inequality in oral health among Iranian adults. Reducing socioeconomic-related inequalities in dental caries should be accompanied by appropriate health promotion policies that focus actions on the fundamental socioeconomic causes of dental disease.


[Influence of estrogen replacement therapy on atherogenic profile in postmenopausal women].
Postmenopausal women have higher risk of cardiovascular events compared with premenopausal. The aim of this clinical study was to evaluate the effect of hormone replacement therapy (HRT) on the atherogenic profile in apparently healthy postmenopausal women. The subjects were 76 healthy postmenopausal women, aged 45 to 59 years, and 15 premenopausal women with regular cycles, aged 40 to 45 years. 63 postmenopausal women completed the study. None of the participating women had a history of hypertension, diabetes mellitus or medications known to affect the cardiovascular system. Twenty seven postmenopausal women received daily 50 micrograms of transdermal estradiol and 100 milligrams of oral progesterone. Thirty six dit not receive HRT. Checkups were preformed at baseline and after six months of treatment, except the group of premenopausal women (only at baseline). Examinations consisted in measurement of body weight, length, waist/hip ratio and plasma levels of biochemical parameters. Estradiol levels were higher among premenopausal women than among treated and non-treated postmenopausal women (83 +/- 78.47 versus 10 +/- 3.99; 12 +/- 4.56 mg/l, p < 0.0001). Levels of serum cholesterol (198 +/- 30.2 versus 236 +/- 33.7; 228 +/- 32.8 mg/dl; p < 0.002), LDL-cholesterol (120 +/- 25 versus 151 +/- 34.2; 144 +/- 31.5 mg/dl, p < 0.02), uric acid (3.98 +/- 0.5 versus 4.6 +/- 1; 4.8 +/- 1 mg/dl, p < 0.004) and homocysteine (9.8 +/- 3.3 versus 13 +/- 3.1; 11 +/- 1.7 mmol/l, p < 0.02) were higher among postmenopausal women than among premenopausal women. The treated women showed higher levels of serum estradiol (49 +/- 34.43 versus 10 +/- 3.99, p < 0.0001) and CRP (0.11 +/- 0.06 vs. 0.24 +/- 0.12 mg/l, p < 0.05), lower waist/hip ratio (0.82 +/- 0.06 versus 0.83 +/- 0.05, p < 0.03), glucose (78 +/- 15.4 versus 85 +/- 14.3 mg/dl, p < 0.03), cholesterol (215 +/- 33.2 versus 236 +/- 33.7 mg/dl, p < 0.03), triglycerides (90 +/- 30.1 versus 106 +/- 47 mg/dl, p < 0.003) and calcium (9.4 +/- 0.4 versus 9.5 +/- 0.4 mg/dl, p < 0.0005) than postmenopausal women without hormone replacement. That HRT may have a favourable effect on atherogenic profile in apparently healthy postmenopausal women.


The long wait for medical excellence.
In October 1997 the book Demanding Medical Excellence: Doctors and Accountability in the Information Age provided a pathbreaking examination of the quality of American health care. It documented rampant medical error and the absence of evidence-based practice, highlighted the potential of electronic health records (EHRs), endorsed what is now known as value purchasing, and showed how patients could exert more control over their care. Although the book suggested that transformational change was imminent, sixteen years later little has changed in some areas (medical error), while in others (evidence-based medicine and population health) change is only now gaining momentum. The exception is technology, where incentives boosted EHR use and the Internet has made a vast array of information available to patients. Paradigm shifts are traumatic, and only recently has intense financial pressure made greater clinical accountability seem less painful than retaining the tradition of untrammeled autonomy. In hearing rooms and hospital hallways, the policy conversation is changing. This shift, though an unavoidable source of anxiety, nonetheless promises a genuine renewal of American medicine.


Emotional outcome after intensive care: literature review.
This paper is a report of a literature review to identify (a) the prevalence of emotional and psychological problems after intensive care, (b) associated factors and (c) interventions that might improve this aspect of recovery. Being a patient in intensive care has been linked to both short- and long-term emotional and psychological consequences. The literature search was conducted during 2006. Relevant journals and databases were searched, i.e. Medline and CINAHL, between the years 1995 and 2006. The search terms were 'anxiety', 'depression', posttraumatic stress', 'posttraumatic stress disorder' and 'intensive care'. Fifteen papers were reviewed representing research studies of anxiety, depression and posttraumatic stress, and seven that represented intensive care follow-up clinics and patient diaries. Being in intensive care can result in significant emotional and psychological problems for a number of patients. For the majority of patients, symptoms of distress will decrease over time but for a number these will endure for some years. Current evidence indicates that emotional problems after intensive care are related to both subjective and objective indicators of a patient's intensive care experience. Evidence suggests some benefit in an early rehabilitation programme, daily sedation withdrawal and the use of patient diaries. However, additional research is required to support such findings. Our understanding of the consequences of intensive care is improving. Psychological care for intensive care patients has lagged behind care for physical problems. We now need to focus on developing and evaluating appropriate interventions to improve psychological outcome in this patient group.


Regional distribution of calcium- and cyclic adenosine 3':5'-monophosphate-regulated protein phosphorylation systems in mammalian brain. I. Particulate systems.
The regional distribution of phosphoproteins whose phosphorylation is regulated either by cyclic AMP or by calcium in combination with calmodulin or phospholipid has been investigated in particulate preparations from rat CNS. About 30 distinct phosphoproteins were observed. These phosphoproteins exhibited widely different patterns of regional distribution. Based upon distribution patterns, we have divided these phosphoproteins into three categories: category A, phosphoproteins found in all parts of the CNS in approximately equal amounts; category B, phosphoproteins which are widely distributed within the CNS but show large regional variations; and category C, phosphoproteins which show a highly restricted regional distribution. We have tentatively interpreted the results on particulate phosphoproteins in the following way: some are present in all or nearly all brain cells, others are present only in certain classes of brain cells, and still others have an even more limited distribution, being present in only a single type of brain cell. The regional distribution of particulate protein kinase activity was also examined. Calcium/calmodulin-dependent protein kinase activity had a marked regional distribution, whereas cyclic AMP-dependent protein kinase activity was more evenly distributed. Calcium/phospholipid-dependent protein kinase activity was barely detectable under the experimental conditions used. This investigation thus demonstrates striking differences in the regional distribution of particulate protein phosphorylation systems in mammalian brain. These regional differences may reflect highly specific functional roles for certain of these protein phosphorylation systems. Similar conclusions concerning cytosolic protein phosphorylation systems are described in the accompanying paper.


Multiplex PCR assays for the detection of clinically relevant antibiotic resistance genes in staphylococci isolated from patients infected after cardiac surgery. The ESPRIT Trial.
Multiresistant staphylococci (82 Staphylococcus aureus and 114 coagulase-negative staphylococci) were characterized by testing with rapid multiplex polymerase chain reaction (PCR) assays for species identification and detection of associated antibiotic resistance genes. These 196 staphylococci were isolated from 149 adult patients who developed wound infection after elective coronary artery bypass grafts and/or valve surgery. The multiplex PCR assays allowed identification of the most common staphylococcal species with S. aureus- and Staphylococcus epidermidis-specific primers as well as the detection of the erythromycin resistance genes ermA, ermB, ermC and msrA, the aminoglycoside resistance gene aac(6')-aph(2"), the oxacillin resistance gene mecA and the penicillin resistance gene blaZ. There was a very good correlation between the genotypic analysis by PCR and the phenotype determined by standard methods of susceptibility testing and identification of staphylococcal species: 100% for erythromycin resistance, 98.0% for gentamicin resistance, 99.0% for oxacillin resistance, 100% for penicillin resistance and 100% for S. aureus and S. epidermidis species identification. This study suggests that the incidence and distribution of the tested clinically relevant antibiotic resistance genes in staphylococci associated with infections after cardiac surgery do not differ from those in strains from other infections. These multiplex PCR assays may be used as diagnostic tools to replace or complement standard methods of susceptibility testing and identification of staphylococci.


Photoacoustic temperature imaging based on multi-wavelength excitation.
Building further upon the high spatial resolution offered by ultrasonic imaging and the high optical contrast yielded by laser excitation of photoacoustic imaging, and exploiting the temperature dependence of photoacoustic signal amplitudes, this paper addresses the question whether the rich information given by multispectral optoacoustic tomography (MSOT) allows to obtain 3D temperature images. Numerical simulations and experimental results are reported on agarose phantoms containing gold nanoparticles and the effects of shadowing, reconstruction flaws, etc. on the accuracy are determined.


Transitions in drinking in adolescent females: evidence from the Missouri adolescent female twin study.
Data from adult community samples indicate that alcohol dependence originates in late adolescence, thus marking that period as a fruitful one for the study of transitions in drinking. Using retrospective data about the timing of alcohol symptom onset from 1799 female twins who participated in the Missouri Adolescent Female Twin Study (MOAFTS), we integrated person-centered (PC) and variable-centered (VC) analyses to study 1-year transitions in alcohol symptomatology. A person-year data set reflecting the onset of 15 alcohol dependence symptoms was constructed, with each year in the young woman's life from the year of her first drink to her current age counting as one observation. Latent class analysis--a PC technique--identified classes of alcohol symptomatology for each person-year, and a lag function was used to identify class membership 1 year earlier. One-year transitions in drinking--a VC analysis--were then studied. Smoking, marijuana use, and conduct problems were consistent promoters of transitions to more severe drinking classes, but depressed affect was not a significant predictor. Parental history of alcohol problems or excessive drinking, particularly maternal history, was a significant predictor of more serious transitions to moderate and dependent problem drinking. These data confirm that even over a 1-year time frame there was considerable change in drinking patterns in this adolescent sample. Smoking was as potent a risk factor as conduct problems in promoting the progression of alcohol symptomatology, and the role of familial factors was found to be influential in transitions at the more severe end of the drinking spectrum.


Azaspirane (N-N-diethyl-8,8-dipropyl-2-azaspiro [4.5] decane-2-propanamine) inhibits human multiple myeloma cell growth in the bone marrow milieu in vitro and in vivo.
Azaspirane (N-N-diethyl-8,8-dipropyl-2-azaspiro [4.5] decane-2-propanamine; trade name, Atiprimod) is an orally bioavailable cationic amphiphilic compound that significantly inhibits production of interleukin 6 (IL-6) and inflammation in rat arthritis and autoimmune animal models. We here characterize the effect of atiprimod on human multiple myeloma (MM) cells. Azaspirane significantly inhibited growth and induced caspase-mediated apoptosis in drug-sensitive and drug-resistant MM cell lines, as well as patient MM cells. IL-6, insulin-like growth factor 1 (IGF-1), or adherence of MM cells to bone marrow stromal cells (BMSCs) did not protect against atiprimod-induced apoptosis. Both conventional (dexamethasone, doxorubicin, melphalan) and novel (arsenic trioxide) agents augment apoptosis induced by atiprimod. Azaspirane inhibits signal transducer activator of transcription 3 (STAT3) and a PI3-K (phosphatidylinositol 3-kinase) target (Akt), but not extracellular signal-regulated kinase 1 and 2 (ERK1/2), inhibits phosphorylation triggered by IL-6, and also inhibits inhibitorkappaBalpha (IkappaBalpha) and nuclear factor kappaB (NFkappaB) p65 phosphorylation triggered by tumor necrosis factor alpha (TNF-alpha). Of importance, azaspirane inhibits both IL-6 and vascular endothelial growth factor (VEGF) secretion in BMSCs triggered by MM cell binding and also inhibits angiogenesis on human umbilical vein cells (HUVECs). Finally, azaspirane demonstrates in vivo antitumor activity against human MM cell growth in severe combined immunodeficient (SCID) mice. These results, therefore, show that azaspirane both induces MM cell apoptosis and inhibits cytokine secretion in the BM milieu, providing the framework for clinical trials to improve patient outcome in MM.


Multidisciplinary management and emerging therapeutic strategies in aneurysmal subarachnoid haemorrhage.
The management of patients with aneurysmal subarachnoid haemorrhage demands expertise to anticipate, recognise, and promptly treat the many neurological and systemic complications. For this reason, these patients are best cared for in high-volume medical centres with multidisciplinary teams and should preferably be treated in a specialised intensive care unit. Endovascular occlusion and surgical clipping provide complementary alternatives for the treatment of aneurysms. Perfusion scans are redefining the way we detect delayed ischaemia as a growing body of evidence indicates that monitoring vessel diameter is insufficient to prevent cerebral infarctions. Statins, endothelin antagonists, and magnesium sulfate infusion are among the novel strategies being tested for neuroprotection and attenuation of vasospasm. The effectiveness of these treatments is supported by strong experimental data and they represent a new generation of therapeutic options developed from the understanding that vasospasm is primarily caused by endothelial dysfunction.


Apoptosis and pro-inflammatory cytokine response of mast cells induced by influenza A viruses.
The pathogenesis of the influenza A virus has been investigated heavily, and both the inflammatory response and apoptosis have been found to have a definitive role in this process. The results of studies performed by the present and other groups have indicated that mast cells may play a role in the severity of the disease. To further investigate cellular responses to influenza A virus infection, apoptosis and inflammatory response were studied in mouse mastocytoma cell line P815. This is the first study to demonstrate that H1N1 (A/WSN/33), H5N1 (A/Chicken/Henan/1/04), and H7N2 (A/Chicken/Hebei/2/02) influenza viruses can induce mast cell apoptosis. They were found to do this mainly through the mitochondria/cytochrome c-mediated intrinsic pathway, and the activation of caspase 8-mediated extrinsic pathway was here found to be weak. Two pro-apoptotic Bcl-2 homology domain 3 (BH3) -only molecules Bim and Puma appeared to be involved in the apoptotic pathways. When virus-induced apoptosis was inhibited in P815 cells using pan-caspase (Z-VAD-fmk) and caspase-9 (Z-LEHD-fmk) inhibitors, the replication of these three subtypes of viruses was suppressed and the secretions of pro-inflammatory cytokines and chemokines, including IL-6, IL-18, TNF-α, and MCP-1, decreased. The results of this study may further understanding of the role of mast cells in host defense and pathogenesis of influenza virus. They may also facilitate the development of novel therapeutic aids against influenza virus infection.


Coach your physicians to care, listen and connect with patients.
Developing strong physician/patient relationships isn't easy when doctors are pressed for time, forced to see more patients and overburdened with paperwork. Yet, successful organizations realize that the physician/patient relationship is the most valuable asset for the hospital or practice. Learn ways to cultivate the relationship and make it grow even stronger.


Inhibition of MKK7-JNK by the TOR signaling pathway regulator-like protein contributes to resistance of HCC cells to TRAIL-induced apoptosis.
The TOR signaling pathway regulator-like (TIPRL) protein, the mammalian ortholog of yeast TIP41, was identified in an expression profiling screen for factors that regulate human liver carcinogenesis. We investigated the role of human TIPRL protein in hepatocellular carcinoma (HCC). We measured the level of TIPRL in HCC and adjacent nontumor tissues from patients. We used small interfering RNAs and zebrafish to study the function of TIPRL. We used annexin V propidium iodide staining and immunoblot analyses to measure apoptosis and activation of apoptotic signaling pathways. We used confocal microscopy, coimmunoprecipitation, and glutathione-S transferase pull-down analyses to determine interactions among mitogen-activated protein kinase kinase 7 (MKK7 or MAP2K7), TIPRL, and the protein phosphatase type 2A (PP2Ac). We studied the effects of TIPRL in tumor xenografts in mice. Levels of TIPRL were higher in HCC tissues and cell lines than nontumor tissues and primary hepatocytes. Knockdown of tiprl expression in zebrafish led to large amounts of apoptosis throughout the embryos. Incubation of HCC cells, but not primary human hepatocytes, with small interfering RNA against TIPRL (siTIPRL) and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) caused prolonged activation (phosphorylation) of MKK7 and c-Jun N-terminal kinase (JNK) and led to apoptosis, indicated by cleavage of procaspase-8,-3 and of poly-(adenosine diphosphate-ribose) polymerase. TIPRL bound to MKK7 and PP2Ac and promoted the interaction between MKK7 and PP2Ac. In mice, injection of HCC xenograft tumors with siTIPRL and TRAIL led to tumor apoptosis and regression. TIPRL is highly up-regulated in human HCC samples and cell lines, compared with noncancerous liver tissues. TIPRL prevents prolonged activation of MKK7 and JNK and TRAIL-induced apoptosis by mediating the interaction between MKK7 and PP2Ac.


Direct anterior total hip arthroplasty in supine position using regular OR table : case series and review of complication and reoperation rate.
Direct anterior approach total hip arthroplasty (DAA- THA) has gained popularity in the last decades due to multiple advantages : reduced blood loss, muscle sparing, reduced pain, reduced dislocation rate, shorter hospital stay and faster recovery. However, initial studies have reported an unacceptable high intra-operative complication rate, especially during the learning curve. The complications and reoperations in a consecutive series of 356 DAA- THA's using a supine positioning on a regular OR table, without femoral hyperextension, were analysed retrospectively. Conclusion : This study could not confirm the previously reported high complication rate in DAA-THA. The supine positioning without femoral hyperextension is a safe technique, little susceptible by the surgeon's learning curve.


Assessment of compliance with prescribed activity by hemiplegic stroke patients after an exercise programme and physical activity education.
Although post-stroke exercise training programmes improve aerobic capacity and functional capacities in the short-term, the impact on exercise performance at home has not been established. To assess compliance with prescribed physical activity by hemiplegic stroke patients. To compare the patients' stated activity with their actual activity. This was a prospective pilot study of nine hemiplegic patients following an exercise training programme. Each patient's activity was measured using an activity monitor (the ActivPAL) before, during and immediately after the programme and then 3months after the end of the programme. The activity actually performed was compared with the levels recommended after a stroke (at least 30minutes of non-stop activity three times a week). Three months after the end of the programme, the patient's stated and actual activities (in terms of frequency and duration) were compared. Activity patterns changed immediately after the end of the programme. Short-term compliance was good for four patients. However, 3months later, only one patient was performing regular activity in accordance with the guidelines. Hence, medium-term compliance was poor. Most patients overestimated both the duration and frequency of their activity sessions. Three months after the end of the supervised training programme, compliance with physical activity guidelines was low. The ActivPAL activity monitor is easy to use and performs satisfactorily. It can be used as a tool for activity assessment and education intervention. The use of an actimeter may increase the efficacy of health education interventions, which promote physical activity.


The nucleotide sequence of the dnaA gene and the first part of the dnaN gene of Escherichia coli K-12.
The nucleotide sequence of the dnaA gene and the first 10% of the dnaN gene was determined. From the nucleotide sequence the amino acid sequence of the dnaA gene product was derived. It is a basic protein of 467 amino acid residues with a molecular weight of 52.5 kD. The expression of the dnaA gene is in the counterclockwise direction like the one of the dnaN gene, for which potential startsites were found.


Reactive benign follicular mucinosis: a report of 2 cases.
We report 2 cases of adolescents who developed follicular mucinosis following cutaneous infections. A 17-year-old adolescent boy was evaluated for a 2-week history of erythematous papules and plaques on his face and neck. One month prior to presentation a culture was taken that was positive for methicillin-sensitive Staphylococcus aureus-associated impetigo. Biopsies from 2 representative lesions demonstrated follicular mucinosis without evidence of folliculotropism or T cell gene rearrangements. A separate case involved a 17-year-old adolescent girl who presented with an edematous plaque on her right preauricular region and scattered erythematous papules and small annular plaques over her face 2 weeks following a herpes simplex virus type 2 (HHV-2) infection. on her face. Biopsy showed follicular mucinosis without evidence of epidermotropism or lymphocyte atypia. There was no herpesvirus cytopathic effect. The first case rapidly responded to an oral prednisone taper and the second case resolved over several weeks without further treatment.


Gonadotropin-releasing hormone: one polypeptide regulates secretion of luteinizing and follicle-stimulating hormones.
A polypeptide isolated from porcine hypothalami stimulates the release of both luteinizing hormone and follicle-stimulating hormone from the pituitaries of several species. This polypeptide has been structurally identified as (pyro)Glu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH(2) and synthesized. The natural and synthetic materials share biological properties. It appears that this peptide represents the hypothalamic hormone regulating the secretion of both luteinizing hormone and follicle-stimulating hormone.


Coupled membrane transporters reduce noise.
Molecular systems are inherently probabilistic and operate in a noisy environment, yet, despite all these uncertainties, molecular functions are surprisingly reliable and robust. The principles used by natural systems to deal with noise are still not well understood, especially in a nonhomogeneous environment where molecules can diffuse across different compartments. In this paper we show that membrane transport mechanisms have very effective properties of noise reduction. In particular, we show that active transport mechanisms (those that can transport against a gradient of concentration by using energy or by means of the concentration gradient of other substances), such as symporters and antiporters, have surprising efficiency in noise reduction, which outperforms passive diffusion mechanisms and are well below Poisson levels. We link our results to the coupled transport of potassium, sodium, and glucose to show that the noise in internal glucose level can be greatly reduced. Our results show that compartmentalization can be a highly effective mechanism of noise reduction and suggests that membrane transport could give this extra benefit, contributing to the emergence of complex compartmentalization in eukaryotes.


High-dose therapy and autologous stem cell transplantation in first relapse for diffuse large B cell lymphoma in the rituximab era: an analysis based on data from the European Blood and Marrow Transplantation Registry.
Autologous stem cell transplantation (ASCT) consolidation remains the treatment of choice for patients with relapsed diffuse large B cell lymphoma. The impact of rituximab combined with chemotherapy in either first- or second-line therapy on the ultimate results of ASCT remains to be determined, however. This study was designed to evaluate the benefit of ASCT in patients achieving a second complete remission after salvage chemotherapy by retrospectively comparing the disease-free survival (DFS) after ASCT for each patient with the duration of the first complete remission (CR1). Between 1990 and 2005, a total of 470 patients who had undergone ASCT and reported to the European Blood and Bone Transplantation Registry with Medical Essential Data Form B information were evaluated. Of these 470 patients, 351 (74%) had not received rituximab before ASCT, and 119 (25%) had received rituximab before ASCT. The median duration of CR1 was 11 months. The median time from diagnosis to ASCT was 24 months. The BEAM protocol was the most frequently used conditioning regimen (67%). After ASCT, the 5-year overall survival was 63% (95% confidence interval, 58%-67%) and 5-year DFS was 48% (95% confidence interval, 43%-53%) for the entire patient population. Statistical analysis showed a significant increase in DFS after ASCT compared with duration of CR1 (median, 51 months versus 11 months; P < .001). This difference was also highly significant for patients with previous exposure to rituximab (median, 10 months versus not reached; P < .001) and for patients who had experienced relapse before 1 year (median, 6 months versus 47 months; P < .001). Our data indicate that ASCT can significantly increase DFS compared with the duration of CR1 in relapsed diffuse large B cell lymphoma and can alter the disease course even in patients with high-risk disease previously treated with rituximab.


SNP-based breeding for broiler resistance to ascites and evaluation of correlated production traits.
The goal of this study was to evaluate marker-assisted selection (MAS) in broiler chickens using previously mapped gene regions associated with ascites syndrome incidence. The second-generation MAS products were assessed for impact on ascites phenotype and whether there were associated changes in important production traits. Previously, we used whole genome resequencing (WGR) to fine-map 28 chromosomal regions as associated with ascites phenotype in our experimental ascites broiler line (Relaxed, REL) based on a hypobaric chamber challenge. Genotypes for single nucleotide polymorphisms (SNPs) in mapped regions on chromosomes 2 and 22, were used for MAS in our REL line. After two generations, birds homozygous for the genotypes associated with resistance for both chromosomal regions were established. The MAS F2 generation was then compared to the REL line for ascites susceptibility and 25 production traits. Selection based on SNPs in the carboxypeptidase Q (CPQ, Gga2) and leucine rich repeat transmembrane neuronal 4 (LRRTM4, Gga22) gene regions resulted in a sex- and simulated altitude- dependent reduction of ascites incidence in two F2 cohorts of the MAS line. Comparisons of the F2 MAS and REL lines for production traits when reared at ambient pressure found no significant negative impacts for feed intake (FI), feed conversion ratio (FCR), or deboned part yields for either sex for two F2 cohorts. There were, however, improvements in the MAS for full-trial body weight gain (BWG), FCR, absolute and relative tender weights, and relative drumstick weight. These results validate the mapping of the 28 chromosomal regions and demonstrate that fine mapping by WGR is an effective strategy for addressing a complex trait; it also stands as the first successful SNP-based selection program against a complex disease trait, such as ascites. The MAS line is comparable and, in some instances, superior, in growth performance to the REL control while being more resistant to ascites. This study indicates that MAS based on WGR can provide significant breeding potential in agricultural systems.


New forceps free injection technique for delivering descemet membrane endothelial keratoplasty preloaded endothelium-in grafts.
To describe a new method for delivering DMEK grafts into the recipient's eye with endothelium inward configuration using a no-forceps injection technique. We retrospectively review 11 patients that underwent DMEK surgery at our institution using a no-forceps injection technique. The graft was preloaded into an intraocular lens (IOL) cartridge and connected to an anterior chamber maintainer (ACM). A 5 ml non luer lock syringe was inserted into the other end of the ACM to create a one-flow system. The cartridge was inserted into the posterior end of an injector, and the graft was successfully delivered into the recipient's eye. Twelve eyes of 11 patients were included. Mean follow-up was 9.16 ± 1.3 months. At baseline, mean best corrected visual acuity (BCVA) was 0.76 ± 0.13 logMAr and mean endothelial cell density (ECD) was 2619.00 ± 115.89 cells/mm2. At follow-up, BCVA significantly improved to 0.22 ± 0.05 logMAR (p = 0.003). Although we observed a significant reduction in ECD at follow-up (1688 ± 182.20, p = 0.002), our patients lost only 35.69 ± 6.36% of endothelial cells. Our technique can help surgeons safely deliver an endothelium-in graft into the recipient's eye. The method doesn't require the use of a forceps, minimizing the risk of endothelial cell loss or graft damage.


Prostaglandin F2(alpha) stimulates growth of skeletal muscle cells via an NFATC2-dependent pathway.
Skeletal muscle growth requires multiple steps to form large multinucleated muscle cells. Molecules that stimulate muscle growth may be therapeutic for muscle loss associated with aging, injury, or disease. However, few factors are known to increase muscle cell size. We demonstrate that prostaglandin F2alpha (PGF2alpha) as well as two analogues augment muscle cell size in vitro. This increased myotube size is not due to PGF2alpha-enhancing cell fusion that initially forms myotubes, but rather to PGF2alpha recruiting the fusion of cells with preexisting multinucleated cells. This growth is mediated through the PGF2alpha receptor (FP receptor). As the FP receptor can increase levels of intracellular calcium, the involvement of the calcium-regulated transcription factor nuclear factor of activated T cells (NFAT) in mediating PGF2alpha-enhanced cell growth was examined. We show that NFAT is activated by PGF2alpha, and the isoform NFATC2 is required for PGF2alpha-induced muscle cell growth and nuclear accretion, demonstrating the first intersection between prostaglandin receptor activation and NFAT signaling. Given this novel role for PGF2alpha in skeletal muscle cell growth, these studies raise caution that extended use of drugs that inhibit PG production, such as nonsteroidal antiinflammatory drugs, may be deleterious for muscle growth.


Fulvalene as a platform for the synthesis of a dimetallic dysprosocenium single-molecule magnet.
The dinucleating fulvalenyl ligand [1,1',3,3'-(C5 t Bu2H2)2]2- (Fvtttt) was used to synthesize the dimetallic dysprosocenium cation [{Dy(η5-Cp*)}2(μ-BH4)(η5:η5-Fvtttt)]+ (3) as the salt of [B(C6F5)4]- (Cp* = C5Me5). Compound [3][B(C6F5)4] was obtained using a method in which the double half-sandwich complex [{Dy(BH4)2(THF)}2(Fvtttt)] (1) was reacted with KCp* to give the double metallocene [{Dy(Cp*)(μ-BH4)}2(Fvtttt)] (2), followed by removal of a bridging borohydride ligand upon addition of [(Et3Si)2(μ-H)][B(C6F5)4]. The dimetallic fulvalenyl complexes 1-3 give rise to single-molecule magnet (SMM) behaviour in zero applied field, with the effective energy barriers of 154(15) cm-1, 252(4) cm-1 and 384(18) cm-1, respectively, revealing a significant improvement in performance across the series. The magnetic properties are interpreted with the aid of ab initio calculations, which show substantial increases in the axiality of the crystal field from 1 to 2 to 3 as a consequence of the increasingly dominant role of the Fvtttt and Cp* ligands, with the barrier height and hysteresis properties being attenuated by the equatorial borohydride ligands. The experimental and theoretical results described in this study furnish a blueprint for the design and synthesis of poly-cationic dysprosocenium SMMs with properties that may surpass those of benchmark systems.


Sexual conflict over parental investment in repeated bouts: negotiation reduces overall care.
Understanding the evolution of parental care is complicated by the occurrence of evolutionary conflicts of interest within the family, variation in the quality and state of family members, and repeated bouts of investment in a family of offspring. As a result, family members are expected to negotiate over care. We present a model for the resolution of sexual conflict in which parents negotiate over repeated bouts of care. Negotiation is mediated by parents deciding at the start of each bout how much care to give on the basis of the state (mass) of offspring, which reflects the amount of care previously received. The evolutionarily stable pattern of care depends on whether the parents care together for the whole family, or each cares alone for part of the divided family. When they care together, they provide less care in the first bout, more in the last bout, and less care overall, resulting in lower parental and offspring fitness. Our results emphasize that negotiation over parental care may occur as a means of avoiding exploitation owing to sexual conflict, even in the absence of variation in the quality of either sex of parent, and lead to a reduction in fitness.


Homer proteins regulate coupling of group I metabotropic glutamate receptors to N-type calcium and M-type potassium channels.
Group I metabotropic glutamate receptors (mGluR1 and 5) couple to intracellular calcium pools by a family of proteins, termed Homer, that cross-link the receptor to inositol trisphosphate receptors. mGluRs also couple to membrane ion channels via G-proteins. The role of Homer proteins in channel modulation was investigated by expressing mGluRs and various forms of Homer in rat superior cervical ganglion (SCG) sympathetic neurons by intranuclear cDNA injection. Expression of cross-linking-capable forms of Homer (Homer 1b, 1c, 2, and 3, termed long forms) occluded group I mGluR-mediated N-type calcium and M-type potassium current modulation. This effect was specific for group I mGluRs. mGluR2 (group II)-mediated inhibition of N-channels was unaltered. Long forms of Homer decreased modulation of N- and M-type currents but did not selectively block distinct G-protein pathways. Short forms of Homer, which cannot self-multimerize (Homer 1a and a Homer 2 C-terminal deletion), did not alter mGluR-ion channel coupling. When coexpressed with long forms of Homer, short forms restored the mGluR1a-mediated calcium current modulation in an apparent dose-dependent manner. Homer 2b induced cell surface clusters of mGluR5 in SCG neurons. Conversely, a uniform distribution was observed when mGluR5 was expressed alone or with Homer short forms. These studies indicate that long and short forms of Homer compete for binding to mGluRs and regulate their coupling to ion channels. In vivo, the immediate early Homer 1a is anticipated to enhance ion channel modulation and to disrupt coupling to releasable intracellular calcium pools. Thus, Homer may regulate the magnitude and predominate signaling output of group I mGluRs.


Cancer Stem Cells in Thyroid Tumors: From the Origin to Metastasis.
Thyroid tumors are extremely heterogeneous varying from almost benign tumors with good prognosis as papillary or follicular tumors, to the undifferentiated ones with severe prognosis. Recently, several models of thyroid carcinogenesis have been described, mostly hypothesizing a major role of the thyroid cancer stem cell (TCSC) population in both cancer initiation and metastasis formation. However, the cellular origin of TCSC is still incompletely understood. Here, we review the principal epigenetic mechanisms relevant to TCSC origin and maintenance in both well-differentiated and anaplastic thyroid tumors. Specifically, we describe the alterations in DNA methylation, histone modifiers, and microRNAs (miRNAs) involved in TCSC survival, focusing on the potential of targeting aberrant epigenetic modifications for developing novel therapeutic approaches. Moreover, we discuss the bidirectional relationship between TCSCs and immune cells. The cells of innate and adaptive response can promote the TCSC-driven tumorigenesis, and conversely, TCSCs may favor the expansion of immune cells with protumorigenic functions. Finally, we evaluate the role of the tumor microenvironment and the complex cross-talk of chemokines, hormones, and cytokines in regulating thyroid tumor initiation, progression, and therapy refractoriness. The re-education of the stromal cells can be an effective strategy to fight thyroid cancer. Dissecting the genetic and epigenetic landscape of TCSCs and their interactions with tumor microenvironment cells is urgently needed to select more appropriate treatment and improve the outcome of patients affected by advanced differentiated and undifferentiated thyroid cancers.


Association of alcohol dehydrogenase genes with alcohol dependence: a comprehensive analysis.
Linkage evidence indicated that gene(s) located on chromosome 4q, in the region of the alcohol dehydrogenase (ADH) genes, affected risk for alcoholism. We genotyped 110 single nucleotide polymorphisms (SNPs) across the seven ADH genes and analyzed their association with alcoholism in a set of families with multiple alcoholic members, using the pedigree disequilibrium test. There was strong evidence that variations in ADH4 are associated with alcoholism: 12 SNPs were significantly associated. The region of strongest association ran from intron 1 to 19.5 kb beyond the 3' end of the gene. Haplotype tag SNPs were selected for the block in the ADH4 gene that provided evidence of association and subsequently used in association analysis; the haplotype was significantly associated with alcoholism (P=0.01) There was weaker evidence that variations in ADH1A and ADH1B might also play a role in modifying risk. Among African-Americans, there was evidence that the ADH1B*3 allele was protective.


Carbapenem therapy for bacteremia due to extended-spectrum-β-lactamase-producing Escherichia coli or Klebsiella pneumoniae: implications of ertapenem susceptibility.
A retrospective study was conducted at two medical centers in Taiwan to evaluate the clinical characteristics, outcomes, and risk factors for mortality among patients treated with a carbapenem for bacteremia caused by extended-spectrum-beta-lactamase (ESBL)-producing organisms. A total of 251 patients with bacteremia caused by ESBL-producing Escherichia coli and Klebsiella pneumoniae isolates treated by a carbapenem were identified. Among these ESBL-producing isolates, rates of susceptibility to ertapenem (MICs ≤ 0.25 μg/ml) were 83.8% and 76.4%, respectively; those to meropenem were 100% and 99.3%, respectively; and those to imipenem were 100% and 97.9%, respectively. There were no significant differences in the critical illness rate (P = 0.1) or sepsis-related mortality rate (P = 0.2) for patients with bacteremia caused by ESBL-producing K. pneumoniae (140 isolates, 55.8%) and E. coli (111 isolates, 44.2%). Multivariate analysis of variables related to sepsis-related mortality revealed that the presence of severe sepsis (odds ratio [OR], 15.9; 95% confidence interval [CI], 5.84 to 43.34; P < 0.001), hospital-onset bacteremia (OR, 4.65; 95% CI, 1.42 to 15.24; P = 0.01), and ertapenem-nonsusceptible isolates (OR, 5.12; 95% CI, 2.04 to 12.88; P = 0.001) were independent risk factors. The patients receiving inappropriate therapy had a higher sepsis-related mortality than those with appropriate therapy (P = 0.002), irrespective of ertapenem, imipenem, or meropenem therapy. Infections due to the ertapenem-susceptible isolates (MICs ≤ 0.25 μg/ml) were associated with a more favorable outcome than those due to ertapenem-nonsusceptible isolates (MICs > 0.25 μg/ml), if treated by a carbapenem. However, the mortality for patients with bacteremic episodes due to isolates with MICs of ≤ 0.5 μg/ml was similar to the mortality for those whose isolates had MICs of >0.5 μg/ml (P = 0.8). Such a finding supports the rationale of the current CLSI 2011 criteria for carbapenems for Enterobacteriaceae.


p53 alpha-Helix mimetics antagonize p53/MDM2 interaction and activate p53.
Overexpression or hyperactivation of MDM2 contributes to functional inactivation of wild-type p53 in nearly 50% of tumors. Inhibition of p53 by MDM2 depends on binding between an NH(2)-terminal (residues 16-28) p53 alpha-helical peptide and a hydrophobic pocket on MDM2, presenting an attractive target for development of inhibitors against tumors expressing wild-type p53. Here we report that novel p53 alpha-helical peptide mimics based on a terphenyl scaffold can inhibit MDM2-p53 binding in vitro and activate p53 in vivo. Several active compounds have been identified that inhibit MDM2-p53 binding in an ELISA assay with IC(50) of 10 to 20 micromol/L and induce p53 accumulation and activation in cell culture at 15 to 40 micromol/L. These results suggest that p53 alpha-helical mimetics based on the terphenyl scaffold may be developed into potent p53 activators.


Assembly of proteins into membranes.
Two pathways for protein assembly into biological membranes have been proposed. The "signal hypothesis" emphasizes the role of specific membrane proteins in binding the growing polypeptide and conducting it into the bilayer during its synthesis. The "membrane-triggered folding" hypothesis emphasizes self-assembly and the role of changing protein conformation during transfer from an aqueous compartment into a membrane. These ideas provide a framework for reviewing recent data on the biogenesis of membrane proteins.


The effect of alpha lipoic acid supplementation on sperm functions in idiopathic asthenozoospermic patients: a case-controlled study.
Alpha-lipoic acid (ALA) is a natural short chain fatty acid containing sulfhydryl groups generated from octanoic acid and cysteine in the mitochondria, and is found in both the aqueous and lipid phases. The present study aimed to assess the efficacy of ALA supplementation in primary infertile males complaining of idiopathic asthenozoospermia. Eighty patients were randomly allocated to treatment group A (n = 40) and control group B (n = 40) groups, receiving daily doses of 600 mg (divided into two daily doses of 300 mg each) of alpha lipoic acid (ALA) or an identical placebo for 90 days. Semen analysis, anthropometric and total antioxidant capacity were analysed and compared before and after treatment. Daily supplementation with ALA improved total motility and progressive motility of the spermatozoa. In the ALA-treated group, sperm motility and progressive motility increased significantly, similarly, the mean percentage of sperm vitality demonstrated a significant increase among the ALA treated group (p < 0.001). Analysis revealed a statistically significant increase in semen volume and sperm concentration in the ALA supplemented group, while abnormal morphology decreased significantly (p < 0.001). ALA supplementation significantly improved sperm parameters and functional tests in group A patients. ALA supplementation in patients with idiopathic asthenozoospermic thus enhanced sperm quality and viability, which could therefore be considered as an adjunct therapy pending further verification of its association and mechanisms involved.


Parathyroid exploration. A review of 125 cases.
Cervical exploration was performed in 106 patients with primary hyperparathyroidism and in 19 patients with chronic renal failure. Initial exploration for primary hyperparathyroidism was successful in 97% of the cases. Single adenomas were found in 84, double adenomas in six, and multiple-gland hyperplasia in 12 patients. Two of the four patients in whom cervical exploration failed were ultimately given the diagnosis of benign familial hypocalciuric hypercalcemia. Thirteen adenomas were ectopic. Preoperative thallium-technetium scans and ultrasound correctly localized only 63% of the adenomas. Only 45% of the ectopic adenomas were correctly localized. All four glands should be examined at initial exploration. Because of the occurrence of double adenomas, contralateral exploration is always indicated regardless of whether an enlarged gland and a normal one are found on the first side. All enlarged parathyroids should be removed when dealing with single or multiple adenomas; subtotal parathyroidectomy (3 1/2 glands) should be performed in multiple-gland hyperplasia. Frozen section confirmation of excised parathyroid tissue is essential. If exploration is unsuccessful, ectopic locations such as the retroesophageal areas, thymus, anterior and posterior mediastinum, carotid sheath, and thyroid lobe must be searched carefully. Preoperative localization studies are not as reliable as an experience parathyroid surgeon.


Genome editing in the fall armyworm, Spodoptera frugiperda: Multiple sgRNA/Cas9 method for identification of knockouts in one generation.
The CRISPR/Cas9 system is an efficient genome editing method that can be used in functional genomics research. The fall armyworm, Spodoptera frugiperda, is a serious agricultural pest that has spread over most of the world. However, very little information is available on functional genomics for this insect. We performed CRISPR/Cas9-mediated site-specific mutagenesis of three target genes: two marker genes [Biogenesis of lysosome-related organelles complex 1 subunit 2 (BLOS2) and tryptophan 2, 3-dioxygenase (TO)], and a developmental gene, E93 (a key ecdysone-induced transcription factor that promotes adult development). The knockouts (KO) of BLOS2, TO and E93 induced translucent mosaic integument, olive eye color, and larval-pupal intermediate phenotypes, respectively. Sequencing RNA isolated from wild-type and E93 KO insects showed that E93 promotes adult development by influencing the expression of the genes coding for transcription factor, Krüppel homolog 1, the pupal specifier, Broad-Complex, serine proteases, and heat shock proteins. Often, gene-edited insects display mosaicism in which only a fraction of the cells are edited as intended, and establishing a homozygous line is both costly and time-consuming. To overcome these limitations, a method to completely KO the target gene in S. frugiperda by injecting the Cas9 protein and multiple sgRNAs targeting one exon of the E93 gene into embryos was developed. Ten percent of the G0 larvae exhibited larval-pupal intermediates. The mutations were confirmed by T7E1 assay, and the mutation frequency was determined as >80%. Complete KO of the E93 gene was achieved in one generation using the multiple sgRNA method, demonstrating a powerful approach to improve genome editing in lepidopteran and other non-model insects.


The effect of a diabetes education, coping skills training, and care intervention on physiological and psychosocial outcomes in black women with type 2 diabetes.
An 11-week culturally relevant group diabetes self-management training (DSMT), coping skills training (CST), and diabetes care intervention was compared to a 10-week usual diabetes education and diabetes care intervention on physiological and psychosocial outcomes in 109 Black women (aged 48 +/- 10 years) with type 2 diabetes in primary care (PC). Strong time effects for hemoglobin A1c improvement were seen in both groups from baseline to 3 months and remained similar at 12 and 24 months (p < .0001). Systolic blood pressure (p =.01) and low-density lipoprotein cholesterol levels (p = .05) improved in both groups from baseline to 24 months. Baseline quality of life ([QOL]; Medical Outcome Study Short Form-36) was low. Social function, role-emotional, and mental health domains increased initially in both groups then declined slightly, with less decline for the experimental group at 12 months. At 24 months, experimental group scores increased. General health (p = .002), vitality (p = .01), role-physical, and bodily pain (p = .02) domains increased in both groups over time. Perceived provider support for diet (p = .0001) and exercise (p = .0001) increased in both groups over time. Diabetes-related emotional distress decreased in the experimental compared to the control group (group x time, p = .01). Findings suggest that both methods of diabetes education combined with care can improve metabolic control, QOL, and perceptions of provider care. CST may further assist in long-term improvements in health outcomes. Behavioral interventions are needed in addition to routine diabetes care, particularly in PC.


FEAR OF FALLING AND ITS RELATIONSHIP WITH QUALITY OF LIFE IN THE ELDERLY ATTENDING A FAMILY MEDICINE CLINIC IN SOUTH-SOUTH NIGERIA.
Aging is linked to increased morbidity, higher fear of falling (FOF), and decreased activity, with attendant hampered of quality of life (QOL) in the elderly. Therefore, this study determined the relationship between Fear of falling and Quality of life among the elderly attending a tertiary hospital in Port Harcourt, Rivers State. This was a hospital-based cross-sectional study where 292 participants were recruited by a systematic sampling method and the data was collected using an interviewer-administered questionnaire (which includes age and gender, the Modified Fall Efficacy scale to assess FOL and the 12-item short health survey form [SF-12] to asses QOL). The SPSS version 26.0 was used to perform the data analysis. The mean age of the respondents was 68.5 years ±6.6 years. More of the participants were females and they were more likely than males to be very fearful of falling (37.7% vs 11.1%, respectively). Females aged 60-69 (36.3%) were most likely to express a fear of falling. The PCS-12 correlated with fear of falling as the scores were very low indicating that respondents had some level of loss of physical function. MCS did not correlate with Fear of falling as the scores were fair in both those with a FOF (MFES< 7.99) and those without a FOF (MFES>8). The observed correlation between FOF and QOL domain scores suggests that Fear of falling should be routinely investigated among the elderly irrespective of reason(s) for encounter.


Diverging metabolic programmes and behaviours during states of starvation, protein malnutrition, and cachexia.
Our evolutionary history is defined, in part, by our ability to survive times of nutrient scarcity. The outcomes of the metabolic and behavioural adaptations during starvation are highly efficient macronutrient allocation, minimization of energy expenditure, and maximized odds of finding food. However, in different contexts, caloric deprivation is met with vastly different physiologic and behavioural responses, which challenge the primacy of energy homeostasis. We conducted a literature review of scientific studies in humans, laboratory animals, and non-laboratory animals that evaluated the physiologic, metabolic, and behavioural responses to fasting, starvation, protein-deficient or essential amino acid-deficient diets, and cachexia. Studies that investigated the changes in ingestive behaviour, locomotor activity, resting metabolic rate, and tissue catabolism were selected as the focus of discussion. Whereas starvation responses prioritize energy balance, both protein malnutrition and cachexia present existential threats that induce unique adaptive programmes, which can exacerbate the caloric insufficiency of undernutrition. We compare and contrast the behavioural and metabolic responses and elucidate the mechanistic pathways that drive state-dependent alterations in energy seeking and partitioning. The evolution of energetically inefficient metabolic and behavioural responses to protein malnutrition and cachexia reveal a hierarchy of metabolic priorities governed by discrete regulatory networks.