Functional integration of non-native carotenoids into chloroplasts by viral-derived expression of capsanthin-capsorubin synthase in Nicotiana benthamiana.
The biosynthesis of leaf carotenoids in Nicotiana benthamiana was altered by forced re-routing of the pathway to the synthesis of capsanthin, a non-native chromoplast-specific xanthophyll, using an RNA viral vector containing capsanthin-capsorubin synthase (Ccs) cDNA. The cDNA encoding Ccs was placed under the transcriptional control of a tobamovirus subgenomic promoter. Leaves from transfected plants expressing Ccs developed an orange phenotype and accumulated high levels of capsanthin (up to 36% of total carotenoids). This phenomenon was associated with thylakoid membrane distortion and reduction of grana stacking. In contrast to the situation prevailing in chromoplasts, capsanthin was not esterified and its increased level was balanced by a concomitant decrease of the major leaf xanthophylls, suggesting an autoregulatory control of chloroplast carotenoid composition. Capsanthin was exclusively recruited into the trimeric and monomeric light-harvesting complexes of photosystem II (PSII) and shown to significantly contribute to the light-harvesting capacity. On a chlorophyll basis, the concentrations of PSI and PSII reaction centres were not modified. This demonstration that higher plant antenna complexes can accommodate non-native carotenoids provides compelling evidence for functional remodelling of photosynthetic membranes toward a better photoreactivity by rational design of the incorporated carotenoid structures.


Catenulispora subtropica sp. nov. and Catenulispora yoronensis sp. nov.
Three strains of actinomycetes that formed spore chains were isolated from a paddy field and forest soil and subjected to a polyphasic taxonomic characterization study. The cell wall peptidoglycan contained ll-diaminopimelic acid and glycine. The major isoprenoid quinones were MK-9(H(8)) and MK-9(H(6)), and the major cellular fatty acids were iso-C(16 : 0) and anteiso-C(17 : 0). Phylogenetic analysis based on 16S rRNA gene sequences showed that these organisms belonged in a monophyletic cluster with members of the genus Catenulispora. According to the physiological, biochemical and chemotaxonomic data, the novel organisms could be differentiated from recognized members of the genus Catenulispora. Based on the phenotypic and genotypic data, the three strains are considered to represent two novel species of the genus Catenulispora, for which the names Catenulispora subtropica sp. nov. for strains TT 99-48(T) (type strain; =NBRC 103395(T)=KCTC 19328(T)) and TT 01-19 (=NBRC 103396), and Catenulispora yoronensis sp. nov. (type strain TT N02-20(T)=NBRC 103397(T)=KCTC 19327(T)) are proposed. An emended description of the genus Catenulispora is given.


Free-energy surfaces for liquid-phase reactions and their use to study the border between concerted and nonconcerted alpha,beta-elimination reactions of esters and thioesters.
Distinguishing between the concerted second-order mechanism for beta-eliminations and nonconcerted mechanisms with discrete carbanion intermediates is very difficult experimentally, but the ability of quantum chemistry to find stationary points of the free-energy surface in liquid-phase solutions, even for complex reagents, provides a new tool for elucidating such mechanisms. Here we use liquid-phase density functional theory calculations to find transition states and intermediates on the free-energy surfaces of four base-initiated alpha,beta-eliminations of acetoxy and mesyloxy esters and their analogous thioesters. The geometries, free energies, and charge distributions of these structures support a stepwise irreversible first-order elimination from a conjugate base (E1cB(I)) mechanism with acetoxy ester 3, acetoxy thioester 4, and mesyloxy thioester 6. However, mesyloxy ester 5, which has an excellent nucleofuge and a less-acidic proton, follows a concerted but asynchronous E2 mechanism with an E1cB-like transition state. The anti transition state is more favorable than the syn one, even for the poorer nucleofuge and more-acidic thioesters. The article includes a general scheme for describing liquid-phase reactions in terms of free-energy surfaces.


Transmission dynamics of lumpy skin disease in Ethiopia.
Lumpy skin disease (LSD) is a severe disease of cattle caused by a Capripoxvirus and often caused epidemics in Ethiopia and many other countries. This study was undertaken to quantify the transmission between animals and to estimate the infection reproduction ratio in a predominantly mixed crop-livestock system and in intensive commercial herd types. The transmission parameters were based on a susceptible-infectious-recovered (SIR) epidemic model with environmental transmission and estimated using generalized linear models. The transmission parameters were estimated using a survival rate of infectious virus in the environment equal to 0·325 per day, a value based on the best-fitting statistical model. The transmission rate parameter between animals was 0·072 (95% CI 0·068-0·076) per day in the crop-livestock production system, whereas this transmission rate in intensive production system was 0·076 (95% CI 0·068-0·085) per day. The reproduction ratio (R) of LSD between animals in the crop-livestock production system was 1·07, whereas it was 1·09 between animals in the intensive production system. The calculated R provides a baseline against which various control options can be assessed for efficacy.


Image segmentation using a texture gradient based watershed transform.
The segmentation of images into meaningful and homogenous regions is a key method for image analysis within applications such as content based retrieval. The watershed transform is a well established tool for the segmentation of images. However, watershed segmentation is often not effective for textured image regions that are perceptually homogeneous. In order to segment such regions properly, the concept of the "texture gradient" is introduced. Texture information and its gradient are extracted using a novel nondecimated form of a complex wavelet transform. A novel marker location algorithm is subsequently used to locate significant homogeneous textured or non textured regions. A marker driven watershed transform is then used to segment the identified regions properly. The combined algorithm produces effective texture and intensity based segmentation for application to content based image retrieval.


A general strategy for the perfluoroalkylation of arenes and arylbromides by using arylboronate esters and [(phen)CuR(F)].
A versatile method for the synthesis of aryl perfluoroalkanes from arenes and aryl bromides is described. Substituted arenes or aryl bromides are converted in situ to an aryl boronate ester that readily undergoes perfluoroalkylation in air with [(phen)CuR(F)]. A broad range of aryl bromide substrates were perfluoroalkylated in good yield for the first time. [(phen)CuCF(3)] is now commercially available and has been prepared on 20 g scale.


Cordycepin Suppresses Expression of Diabetes Regulating Genes by Inhibition of Lipopolysaccharide-induced Inflammation in Macrophages.
It has been recently noticed that type 2 diabetes (T2D), one of the most common metabolic diseases, causes a chronic low-grade inflammation and activation of the innate immune system that are closely involved in the pathogenesis of T2D. Cordyceps militaris, a traditional medicinal mushroom, produces a component compound, cordycepin (3'-deoxyadenosine). Cordycepin has been known to have many pharmacological activities including immunological stimulating, anti-cancer, and anti-infection activities. The molecular mechanisms of cordycepin in T2D are not clear. In the present study, we tested the role of cordycepin on the anti-diabetic effect and anti-inflammatory cascades in LPS-stimulated RAW 264.7 cells. We confirmed the levels of diabetes regulating genes mRNA and protein of cytokines through RT-PCR and western blot analysis and followed by FACS analysis for the surface molecules. Cordycepin inhibited the production of NO and pro-inflammatory cytokines such as IL-1beta, IL-6, and TNF-alpha in LPS-activated macrophages via suppressing protein expression of pro-inflammatory mediators. T2D regulating genes such as 11beta-HSD1 and PPARgamma were decreased as well as expression of co-stimulatory molecules such as ICAM-1 and B7-1/-2 were also decreased with the increment of its concentration. In accordance with suppressed pro-inflammatory cytokine production lead to inhibition of diabetic regulating genes in activated macrophages. Cordycepin suppressed NF-kappaB activation in LPS-activated macrophages. Based on these observations, cordycepin suppressed T2D regulating genes through the inactivation of NF-kappaB dependent inflammatory responses and suggesting that cordycepin will provide potential use as an immunomodulatory agent for treating immunological diseases.


Genetic variability of the ABCC2 gene and clinical outcomes in pancreatic cancer patients.
Pancreatic ductal adenocarcinoma (PDAC) has an extremely poor prognosis, caused by various factors, such as the aggressiveness of the disease, the limited therapeutic options and the lack of early detection and risk markers. The ATP binding cassette subfamily C member 2 (ABCC2) protein plays a critical role in response to various drugs and is differentially expressed in gemcitabine sensitive and resistant cells. Moreover, single nucleotide polymorphisms (SNPs) in the gene have been associated with differential outcomes and prognosis in several tumour types. The aim of this study was to investigate the possible association between SNPs in the ABCC2 gene and overall survival (OS) in PDAC patients. We analysed 12 polymorphisms, including tagging-SNPs covering all the genetic variability of the ABCC2 gene and genotyped them in 1415 PDAC patients collected within the Pancreatic Disease ReseArch (PANDoRA) consortium. We tested the association between ABCC2 SNPs and PDAC OS using Cox proportional hazard models. We analysed PDAC patients dividing them by stage and observed that the minor alleles of three SNPs showed an association with worse OS [rs3740067: hazard ratio (HR) = 3.29, 95% confidence interval (CI) = 1.56-6.97, P = 0.002; rs3740073: HR = 3.11, 95% CI = 1.52-6.38, P = 0.002 and rs717620: HR = 2.90, 95% CI = 1.41-5.95, P = 0.004, respectively] in stage I patients. In patients with more advanced PDAC, we did not observe any statistically significant association. Our results suggest that rs3740067, rs3740073 and rs717620 could be promising prognostic markers in stage I PDAC patients.


Nanoengineered Colloidal Inks for 3D Bioprinting.
Nanoengineered hydrogels offer the potential to design shear-thinning bioinks for three-dimensional (3D) bioprinting. Here, we have synthesized colloidal bioinks composed of disk-shaped two-dimensional (2D) nanosilicates (Laponite) and poly(ethylene glycol) (PEG). The addition of Laponite reinforces the PEG network and increases viscosity, storage modulus, and network stability. PEG-Laponite hydrogels display shear-thinning and self-recovery characteristics due to rapid internal phase rearrangement. As a result, a range of complex patterns can be printed using PEG-Laponite bioinks. The 3D bioprinted structure has similar mechanical properties compared to the as-casted structure. In addition, encapsulated cells within the PEG-Laponite bioink show high viability after bioprinting. Overall, this study introduces a new class of PEG-Laponite colloidal inks for bioprinting and cell delivery.


[Factors influencing the pulse character of RNA elongation in vitro by E. coli RNA polymerase].
Pause location along primary structure of two RNA fragments each 200 nucleotide residues in the length synthesized from A1 promoters of T7 phage DNA and delta D111 T7 phage DNA was analyzed. No correlation between the location of pauses and GC-rich or self complementary regions of RNA were found. The location of pauses does not change upon the variation of the temperature or ionic strength. Concurrent variation of all four NTP concentrations also did not influence pausing pattern. However the distribution of pauses depends highly on the ratio of the individual substrate concentrations. Substitution of GTP by ITP changes the pausing pattern completely. Inorganic pyrophosphate (PPi) of inhibits RNA elongation preferentially in the regions: NAUN, CGUAG. The study of PPi action on RNA terminated with 3' OCH3-NMP suggest that the sequence-specific inhibition of RNA elongation may be a result of pyrophosphorolysis of terminal nucleotide residues of RNA. It was proposed that the pulse character of RNA elongation stems rather from differences in the kinetic constants of nucleotides attachment and pyrophosphorolysis from the 3'-termini of RNA than by termination signals encoded in the primary structure of DNA. The stable location of pauses in certain short oligonucleotides: AUG, AUU, AAU and some others is in favour of the hypothesis.


Interpersonal rivalries, gender and the intellectual and scientific making of psychoanalysis in 1940s Britain.
This article examines the 1940s debates regarding the status and professional orthodoxy of psychoanalysis following Sigmund Freud's death, by exploring the Anna Freud-Melanie Klein Controversial Discussions in the British Psychoanalytical Society. Focusing on the work of now-forgotten analysts Melitta Schmideberg and Edward Glover, and on their relationship with Klein and her supporters, the article reveals how these neglected, yet important, debates were complicated by interpersonal and professional ties, processes of the professionalization, and changing gender norms. Although historians of psychoanalysis have not ignored the jealousies, resentments, and complex relationships between psychoanalysts, these scholars often continue to view these as separate from the processes of creating science. Here, instead, I view the personal and the intellectual in tandem, thus challenging the divide between scientific reason and affect. Rather than imposing a separation between the scientific and the personal, I suggest that we should explore how historical actors negotiated the divide themselves. Indeed, I demonstrate that the study of interpersonal contexts is an invaluable tool for understanding the development of psychological disciplines. (PsycINFO Database Record


Protective immunity to Japanese encephalitis virus associated with anti-NS1 antibodies in a mouse model.
Japanese encephalitis virus (JEV) is a major mosquito-borne pathogen that causes viral encephalitis throughout Asia. Vaccination with an inactive JEV particle or attenuated virus is an efficient preventative measure for controlling infection. Flavivirus NS1 protein is a glycoprotein secreted during viral replication that plays multiple roles in the viral life cycle and pathogenesis. Utilizing JEV NS1 as an antigen in viral vectors induces a limited protective immune response against infection. Previous studies using E. coli-expressed JEV NS1 to immunize mice induced protection against lethal challenge; however, the protection mechanism through cellular and humoral immune responses was not described. JEV NS1 was expressed in and purified from Drosophila S2 cells in a native glycosylated multimeric form, which induced T-cell and antibody responses in immunized C3H/HeN mice. Mice vaccinated with 1 μg NS1 with or without water-in-oil adjuvant were partially protected against viral challenge and higher protection was observed in mice with higher antibody titers. IgG1 was preferentially elicited by an adjuvanted NS1 protein, whereas a larger load of IFN-γ was produced in splenocytes from mice immunized with aqueous NS1. Mice that passively received anti-NS1 mouse polyclonal immune sera were protected, and this phenomenon was dose-dependent, whereas protection was low or delayed after the passive transfer of anti-NS1 MAbs. The purified NS1 subunit induced protective immunity in relation with anti-NS1 IgG1 antibodies. NS1 protein efficiently stimulated Th1-cell proliferation and IFN-γ production. Protection against lethal challenge was elicited by passive transfer of anti-NS1 antisera, suggesting that anti-NS1 antibodies play a substantial role in anti-viral immunity.


Mechanism of inactivation of enzyme proteins by ultraviolet light.
Some quantum yields for the destruction of amino acids have been determined. The inactivation of the enzymes hymotrypsin, lysozyme, ribonuclease, and trypsin by ultraviolet light can be accounted for quantitatively by summing the products of (i) the probability that light is absorbed by a given amino acid residue, epsilon(4), and (ii) the probability that absorbed light induces a chemical change, with a quantum efficiency phi(4), in the residue. The principal residues involved are cystyl and tryptophanyl. Peptide bond rupture is not important. Analysis of inactivated enzymes verifies the assumption of the existence of several inactivation mechanisms.


High-yield isolation of functionally competent endosomes from mouse lymphocytes.
A discontinuous-sucrose-gradient procedure for isolating endosomes from mouse lymphoma cells has been developed. After centrifugation, most organelles (especially mitochondria and lysosomes) are recovered in the denser fractions of the gradient, whereas a mixture of plasma membrane and endosomes is present at lighter densities. The endosome recovery in this fraction can be increased (by 100%) by (a) a mild trypsin treatment of the postnuclear supernatant and (b) loading the cell endosomes with a saturating concentration of low-density lipoproteins. Removal of the plasma-membrane contamination was achieved by preincubating the cells with a gold-ricin complex at 4 degrees C. On centrifugation, the gold-loaded membranes sediment to the bottom of the gradient. The endosome preparation isolated by these procedures is less than 6% contaminated by other organelles and contains 42% of internalized 125I-transferrin. We show that these isolated endosomes are functional, as displayed by their ability to fuse and to acidify in a cell-free system. Endosome fusion was studied by a new assay based on the use of fluorescence resonance energy transfer. This fusion is dependent on ATP and on a cytosolic, thermoresistant but trypsin- and N-ethylmaleimide-sensitive, protein factor. Early endosomes fuse more actively among themselves than with late-endocytic vesicles, and they fuse only slowly with plasma-membrane vesicles.


Training and supporting pharmacists to supply progestogen-only emergency contraception.
To describe and evaluate the training and support provided to the first cohort of community pharmacists to supply progestogen-only emergency contraception (POEC) under a Patient Group Direction (PGD) in Lambeth, Southwark and Lewisham, London. The study comprised (a) a systematic analysis of written and oral data from pharmacists before and during training, and at 5 and 13-14 months after launch; (b) analysis of telephone calls to clinical support and (c) analysis of written pharmacy records. A total of 20/22 pharmacists in the first training cohort; 6/23 pharmacists who applied but were not accepted were also followed up. A formal course with role-play was a successful training method, and the course also served as a team-building exercise. Subsequent interviews demonstrated that pharmacists had understood the concept of client confidentiality and gained confidence over time in the use of the PGD. The on-call consultants received 152 calls in the first 12 months of the scheme. Over 80% of the calls concerned clinical criteria (notably including 22% that were queries about oral contraceptives). Frequency ranged from one to eight calls per week with 28% made at weekends. In over half (60%) of the calls the pharmacist was subsequently able to make a supply. Queries over client management resulted in several changes in the protocol. The primary expressed concern for all pharmacists at all time points was how clients might 'misuse' or 'abuse' the service, and this remained a concern despite the fact that it also applies to other routes of supply of POEC. However, the PGD cohort was more positive on local benefits than pharmacists who were not selected. Training and support have enabled this often-underused group of professionals to participate in an extended reproductive health service. Mobile phones are an essential support tool.


Suicidal behavior runs in families. A controlled family study of adolescent suicide victims.
While previous studies have shown an increased rate of suicidal behavior in the relatives of suicide victims, it is unclear if this is attributable merely to increased familial rates of psychiatric disorders. Therefore, we conducted a family study of adolescent suicide victims (suicide probands) and community control probands (controls) to determine if the rates of suicidal behavior were higher in the relatives of adolescent suicide probands even after adjusting for differences in the familial rates of psychiatric disorders. The relatives of 58 adolescent suicide probands and 55 demographically similar controls underwent assessment for Axis I and II psychiatric disorders, lifetime history of aggression, and history of suicidal behavior (attempts and completions) using a combination of family study and family history approaches. The rate of suicide attempts was increased in the first-degree relatives of suicide probands compared with the relatives of controls, even after adjusting for differences in rates of proband and familial Axis I and II disorders (odds ratio, 4.3; 95% confidence intervals, 1.1-16.6). On the other hand, the excess rate of suicidal ideation found in the relatives of suicide probands was explained by increased familial rates of psychiatric disorders. Among suicide probands, higher ratings of aggression were associated with higher familial loading for suicide attempts. Liability to suicidal behavior might be familially transmitted as a trait independent of Axis I and II disorders. The transmitted spectrum of suicidal behavior includes attempts and completions, but not ideation, and the transmission of suicidal behavior and aggression are related.


An in vitro model of Chlamydia trachomatis infection in the regenerative phase of the human endometrial cycle.
An in vitro model of the regenerative phase of the human endometrial cycle was developed in order to study the growth of Chlamydia trachomatis during the period following menses. Glandular epithelial fragments were prepared from curettings of endometria and explanted onto coated substrata. Epithelial cells migrated rapidly from the explant in a fashion which closely mimicked the regeneration of the surface epithelium after menses. The cultures were then experimentally infected with C. trachomatis serotype E at various times during formation of the outgrowth. Chlamydial inclusions developed both within the explants and in the outgrowing epithelial sheets. They were also found in isolated epithelial and non-epithelial cells. However, the most striking feature of chlamydial inclusion development within these cultures was the tendency for inclusions to be located in cells at the periphery of the epithelial sheets. This was partly due to the failure of the cells within the sheets to bind chlamydiae after centrifugation of the organisms onto the culture and partly due to a phenomenon similar to phagokinesis. During this process infectious chlamydial particles were cleared from the substratum by migrating cells with free motile edges, which occasionally led to internalization and inclusion development within these cells.


Microbial ecology and activities in the rumen: part 1.
This review describes the progress which has been made during the last 10 to 15 years in the field of rumen microbiology. It is basically an account of new discoveries in the bacteriology, protozoology, biochemistry, and ecology of the rumen microbial population. As such it covers a wide range of subjects including the isolation and properties of methanogenic bacteria, the role of rumen phycomycete fungi, anaerobic energy conservation, and general metabolic aspects of rumen microorganisms. It also attempts, however, to describe and develop new concepts in rumen microbiology. These consist principally of interactions of the microbemicrobe, microbe-food and microbe-host types, and represent the main areas of recent advance in our understanding of the rumen ecosystem. The development of experimental techniques such as chemostat culture and scanning electron microscopy are shown to have been instrumental in progress in these areas. The paper is concluded with an assessment of our present knowledge of the rumen fermentation, based on the degree of success of experiments with gnotobiotic ruminants inoculated with defined flora and in mathematical modeling of the fermentation. The efficacy of chemical manipulation of the fermentation in ruminant is also discussed in this light.


The Vast and Varied Global Burden of Norovirus: Prospects for Prevention and Control.
Globally, norovirus is associated with approximately one-fifth of all diarrhea cases, with similar prevalence in both children and adults, and is estimated to cause over 200,000 deaths annually in developing countries. Norovirus is an important pathogen in a number of high-priority domains: it is the most common cause of diarrheal episodes globally, the principal cause of foodborne disease outbreaks in the United States, a key health care-acquired infection, a common cause of travel-associated diarrhea, and a bane for deployed military troops. Partly as a result of this ubiquity and burden across a range of different populations, identifying target groups and strategies for intervention has been challenging. And, on top of the breadth of this public health problem, there remain important gaps in scientific knowledge regarding norovirus, especially with respect to disease in low-income settings. Many pathogens can cause acute gastroenteritis. Historically, rotavirus was the most common cause of severe disease in young children globally. Now, vaccines are available for rotavirus and are universally recommended by the World Health Organization. In countries with effective rotavirus vaccination programs, disease due to that pathogen has decreased markedly, but norovirus persists and is now the most common cause of pediatric gastroenteritis requiring medical attention. However, the data supporting the precise role of norovirus in low- and middle-income settings are sparse. With vaccines in the pipeline, addressing these and other important knowledge gaps is increasingly pressing. We assembled an expert group to assess the evidence for the global burden of norovirus and to consider the prospects for norovirus vaccine development. The group assessed the evidence in the areas of burden of disease, epidemiology, diagnostics, disease attribution, acquired immunity, and innate susceptibility, and the group considered how to bring norovirus vaccines from their current state of development to a viable product that will benefit global health.


Does Type of Social Support Influence Medication Adherence Among South African Adolescents Living With HIV? A Quantitative Analysis of Pilot Data.
Adolescents living with HIV often experience worse medication adherence than older and younger individuals living with HIV. One factor that may mitigate poor adherence is social support. However, few studies have examined the influence of specific social support constructs on HIV outcomes like medication adherence. In response to this gap, we performed bivariate analyses using data from 104 adolescents living with HIV in Cape Town, South Africa to understand the relationship between three types of social support (emotional, informational, and instrumental support) and four HIV-related outcomes: adherence self-efficacy, last recorded CD4+ count, last recorded viral load, and adherence to clinic appointments in the past year. We found small positive associations between two social support types (emotional and informational support) and adherence self-efficacy, with no other statistically significant associations. In light of our small sample size, future studies should seek to understand the differential effects of specific types of social support on HIV-related outcomes to better inform interventions for adolescents living with HIV.


Effect of Ringer infusion on ischemic acute renal failure: caution on interpreting the results of short-term studies.
The effect of a 4.5 mL/h Ringer infusion on the recovery from a unilateral 40-min renal artery occlusion was investigated in Sprague-Dawley rats. The inulin clearance measured in the experimental kidney 24 and 48 h after the insult in control animals that did not receive the Ringer infusion was 0.14 +/- 0.10 (mean +/- SE) and 0.11 +/- 0.05 mL/min, respectively. In animals that received 24 h of Ringer infusion begun at the time of the renal artery occlusion the inulin clearance was 0.81 +/- 0.07 mL/min, a value significantly higher than either of the control groups (p less than .05). If, however, the Ringer infusion was stopped at 24 h and the inulin clearance measured at 48 h, it had decreased significantly (0.27 +/- 0.09 mL/min) and was no longer greater than the control groups. Similarly, if the infusion was continued for 48 h there was no longer a significant difference between the inulin clearance (0.37 +/- 0.11), when compared with 48 h of no infusion (0.11 +/- 0.05). The histology of the different groups corresponded with the functional data. We conclude that 24 h of Ringer infusion leads to functional and histological protection when measured at 24 h; however, if measured at 48 h, protection is no longer evident. These studies suggest that caution should be exercised in extrapolating from the results of protective maneuvers in ischemic acute renal failure investigated by short-term studies.


Dental Extractions Before Radiation Therapy and the Risk of Osteoradionecrosis in Patients With Head and Neck Cancer.
Patients with head and neck cancer undergo extraction of teeth with poor prognoses to minimize post-radiation therapy (RT) extractions, which are known to cause osteoradionecrosis (ORN). However, many patients are required to start RT before the extraction sites are completely healed. The role of pre-RT extractions in the development of ORN has been disputed in literature. To determine whether the timing of pre-RT dental extractions is associated with ORN development in patients with head and neck cancer. This retrospective cohort study was conducted at a single institution (Princess Margaret Cancer Centre, Toronto, Canada) between January 1, 2011, and January 1, 2018, and included 879 patients with head and neck cancer who underwent pre-RT dental extractions before curative RT of 45 Gy or greater. Patient demographic information and clinical characteristics (eg, primary cancer site, nodal involvement, chemotherapy, smoking status, dental pathology) were considered. Data analyses were performed from July to December 2022. Timing (number of days) from dental extractions to RT start date and pre-RT extractions categorized as healed, minor bone spicules (MBS), or ORN. The study population consisted of 879 patients with a median (range) age of 62 (20-96) years, with 685 men (78%) and 194 women (22%). Of these, 847 (96.3%) healed from pre-RT dental extractions, 16 (1.8%) developed MBS, and 16 (1.8%) developed ORN. The median (range) time in number of days from pre-RT extraction(s) to start of RT was 9 (0-98) days in the healed cohort, 6 (3-23) days in the MBS cohort, and 6 (0-12) days in the ORN cohort. There was a large difference in the timing of pre-RT extractions between the healed and the MBS cohorts (mean 11.9 vs 7.4 days to radiation; difference 4.4; 95% CI, 1.5-7.3), and the healed and the ORN cohorts (mean 11.9 vs 7.1 days; difference 4.8 days; 95% CI, 2.6-7.1). The findings of this retrospective cohort study suggest that there was an important association between the timing of pre-RT dental extractions and ORN when extractions occurred within 7 days of the RT start date. Despite this, ORN after pre-RT extractions is relatively rare. These findings indicate that patients with head and neck cancer who are to undergo RT should not delay treatment for extractions when it might compromise oncologic control.


Magnetic nanoparticles as contrast agents in biomedical imaging: recent advances in iron- and manganese-based magnetic nanoparticles.
Improvements in diagnostic measures for biomedical applications have been investigated in various studies for better interpretations of biological abnormalities and several medical conditions. The use of imaging techniques, such as magnetic resonance imaging (MRI), is widespread and becoming a standard procedure for such specialized applications. A major avenue being studied in MRI is the use of magnetic nanoparticles (NPs) as contrast agents (CAs). Among various approaches, current research also incorporates use of superparamagnetic iron oxide NPs and manganese-based NPs with biocompatible coatings for improved stability and reduced biodegradation when exposed to a biological environment. In this review, recent advances with these types of magnetic NPs and their potential use as CAs in MRI are reported, as well as new insights into the selectivity and cellular transport mechanism that occurs following injection.


Sensitivity of intracellular bacteriophage lambda to colicin CA42-E2.
Treatment of Escherichia coli K-12 infected by lambda CIts857 with colicin CA42-E2 resulted in partial inhibition of the infectious process. Uninfected bacteria were killed by colicin with a probability of about five times that with which similarly treated lambda-infected bacteria lose plaque-forming ability. The lambda deoxyribonucleic acid (DNA), when present in a bacterial cell either as the replicating DNA of infectious phage or as the nonreplicating DNA of superinfecting phage, was degraded to acid-soluble material after colicin treatment. Analysis of the intermediates of DNA breakdown has revealed that degradation of the DNA to acid-soluble material is preceded by endonucleolytic fragmentation of the chromosome at a limited number of sites. This is the same mechanism of degradation previously observed for E. coli DNA after colicin treatment.


Pollutant sources investigation and remedial strategies development for the Kaoping River Basin, Taiwan.
The Kaoping River Basin, located in southern Taiwan, flows through approximately 171 km and drains towards the South Taiwan Strait. It is the largest and the most intensively used river basin in Taiwan. Based on the results from the pollutant sources investigation and water quality analysis, the main water pollution sources of the Kaoping River were livestock wastewater from hog farms, municipal wastewater, industrial wastewater, leachate from riverbank landfills, and non-point source (NPS) pollutants from agricultural areas in the upper catchment. Concern about the deteriorating condition of the river led the Government of Taiwan to amend the relevant legislation and strengthen the enforcement of the discharge regulations to effectively manage the river and control the pollution. The following remedial strategies have been taken to improve the river water quality since 2001: (1) hog ban in the upper catchment of the Kaoping River Basin, thus, 510 thousand hogs have been removed/relocated; (2) removal of riverbank landfills; (3) enforcement of the industrial wastewater discharge standards; (4) sewer system construction in five cities along the river corridor; (5) application of best management practices for NPS pollutant control; (6) application of natural wastewater treatment systems (e.g. land treatment, constructed wetland, overland flow, riverbank sedimentation/aeration pond) for domestic wastewater treatment in rural areas; and (7) construction of the watershed geographical information system (GIS) and real time water quality monitoring system to effectively monitor and manage the watershed. Recent water quality investigation results indicate that the biochemical oxygen demand (BOD) and nutrient loadings to the Kaoping River have been significantly reduced and the water quality has been improved after the implementation of the remedial strategies described above. Results and experience obtained from this study will be helpful in designing the watershed management strategies for other similar river basins.


Short acidic peptides isolated from wheat sprout chromatin and involved in the control of cell proliferation. Characterization by infrared spectroscopy and mass spectrometry.
Low molecular weight peptides were isolated from the chromatin of wheat sprouts. Following gel filtration the peptide fraction shows a sharp inhibiting activity on the growth of HeLa cancer cells. Infrared (IR) spectroscopy and mass spectrometry have been utilized to characterize the wheat sprout peptides in an attempt to recognize the peptide sequence involved in the control of cell growth. The quantitative presence of a peptide with MH+=572 appears proportional to the cell growth inhibition activity. This compound has been subjected to extensive mass spectrometry analysis. The automatic computational analysis of the ions of second, third and fourth generations indicate a peptide sequence, AcHis-Asp-Ser-Glu-, that binds at the C-terminal a molecule of ethanolamine. Moreover, the results show that some sequences of the wheat sprout peptide family are present in the peptide fractions isolated from several other tissues, thus supporting the hypothesis of ubiquitous regulatory peptides.


Deep peroneal nerve paresis in a runner caused by ganglion at capitulum peronei. Case report and review of the literature.
Although lateral popliteal sciatic nerve damage is not one of the commonest diseases in the general population, it is quite frequent among athletes. Several physiopathologic mechanisms have been thought to bring about this damage in athletes. Soft tissue ganglions with neurological involvement of the lateral popliteal sciatic nerve or its terminal rami are in differential diagnosis with several lesions of this area, as direct or indirect trauma, subcutaneous rupture of anterior tibialis muscle and long peroneal muscle, disc hernia, intraspinal tumor, anterior tarsal tunnel syndrome, cysts, neurofibroma, baker's cyst, vascular claudication, stenosing or inflammatory pathology of 2(nd) motoneuron, antimicrobial agents for urinary tract infection (nitrofurnantoin). The authors report the case of a 34-year-old amateur athlete with a recent paralysis of the hallux extensor, paresis of the toe extensor and hyposthenia of the tibialis anterior. The patient had been suffering from episodes of lumbalgia for a long time. He was sent to us because neurological damage due to disc herniation was suspected. Electromyography, sonography, and CT showed peripheral compression of the deep peroneal nerve caused by a mucous cyst at the capitulum peronei, a ''rare'' condition. The patient underwent surgery to excise the cyst, which led to the rapid resolution of the nerve deficit shown by clinical and electromyographical tests. A meticulous anamnesis and accurate objective examination, followed by specific tests (radiographs, sonography, and possibly CT scan) generally enable a correct diagnosis to be made. If diagnosis and therapy are carried out correctly, and without delay, symptoms quickly resolve and the nerve deficit progressively regresses.


Diagnostic value of MPO in patients admitted for suspected acute coronary syndrome--a study of adult in Mostar, Bosnia and Herzegovina.
We assessed the diagnostic efficacy of plasma Myeloperoxidasis (MPO) alone or in combination with cardiac troponin I (cTnI) for detecting ACS in patients presenting with chest pain initiating within 24 h before the hospital admission. In this prospective cohort study were included all respondents who have visited outpatient clinic of internal diseases, University Hospital Mostar because of chest pain and suspected acute coronary syndrome within 24 h of the onset of the period of 6 months and the total sample consisted of 114 patients. Troponin and myeloperoxidase were significantly positively correlated at the beginning of treatment, myeloperoxidase was significantly positively associated with adverse cardiovascular events during hospitalization and myocardial infarction (p < 0.05), with the regression analysis did not show a significant predictor in the development of myocardial infarction (p > 0.05). Sensitivity of myeloperoxidase as a valid test detection of myocardial infarction at baseline was 0.15 and specificity was 0.85, suggesting good diagnostic value usable in the clinical practice.


Mucin-type O-glycosylation is critical for vascular integrity.
Vascular endothelial cells, in addition to many other mammalian cell types, express proteins that are highly modified with mucin-type O-glycosylation, a specific type of glycosylation that begins with the addition of an N-acetylgalactosamine moiety to serine or threonine residues within the peptide backbone. Recently, it has become evident that O-glycosylation governs the separation of blood and lymphatic vessels throughout life and plays a critical role in maintaining vascular integrity in specific tissues such as the brain and lymph node. This mini-review seeks to highlight some of these recent advances regarding in vivo functions of mucin-type O-glycans.


Early weaning is beneficial to prevent atopic dermatitis occurrence in young children.
Epidemiological data on infant feeding practices and allergic diseases are controversial. The purpose of this study was to explore the association of early weaning with the occurrence of atopic dermatitis (AD). We conducted a matched case-control study on incident physician-diagnosed AD in early childhood including 451 cases and 451 controls. Data on several factors, including feeding practices, were collected through an interviewer-administered questionnaire. Odds ratios (OR) and the corresponding 95% confidence intervals (CIs) were estimated through logistic regression models, conditioned on study center, age, sex, and period of interview, and adjusted for potential confounders. Early weaning, defined as the introduction of solid foods at 4 or 5 months of age, was inversely related to the risk of AD, with children weaned at 4 months having lower AD risk (OR = 0.41, 95% CI, 0.20-0.87) compared to those exclusively breastfed. Similar results were observed for weaning started at 5 months of age (OR = 0.39, 95% CI, 0.18-0.83). This association persisted when children with and without family history of allergy were considered separately. Prolonged partial breastfeeding (breastmilk plus milk formulas) was not associated with AD. Consistently, the introduction of a high number of different solid foods reduced the risk of AD (P trend = 0.02 at 4 months of age and P trend = 0.04 at 5 months). Our data provide evidence against the preventing role of prolonged exclusive (but not partial) breastfeeding in AD occurrence and confirm recent results indicating a beneficial role of early weaning in AD.


Local Firework Restrictions and Ocular Trauma.
Fireworks can cause vision-threatening injuries, but the association of local legislation with the mitigation of these injuries is unclear. To evaluate the odds of firework-related ocular trauma among residents of areas where fireworks are permitted vs banned. This case-control study was conducted at a level 1 trauma center in Seattle, Washington, among 230 patients presenting with ocular trauma in the 2 weeks surrounding the Independence Day holiday, spanning June 28 to July 11, over an 8-year period (2016-2022). Firework ban status of patient residence. Odds of firework-related injuries among residents of areas where fireworks are legal vs where they are banned, calculated as odds ratios (ORs) and 95% CIs. Of 230 consultations for ocular trauma during the study period, 94 patients (mean [SD] age, 25 [14] years; 86 male patients [92%]) sustained firework-related injuries, and 136 (mean [SD] age, 43 [23] years; 104 male patients [77%]) sustained non-firework-related injuries. The odds of firework-related ocular trauma were higher among those living in an area where fireworks were legal compared with those living in an area where fireworks were banned (OR, 2.0 [95% CI, 1.2-3.5]; P = .01). In addition, the odds of firework injuries were higher for patients younger than 18 years (OR, 3.1 [95% CI, 1.7-5.8]; P < .001) and for male patients (OR, 3.3 [95% CI, 1.5-7.1]; P = .004). Firework injuries were more likely to be vision threatening (54 of 94 [57%]) compared with non-firework-related injuries (54 of 136 [40%]; OR, 2.1 [95% CI, 1.2-3.5]; P = .01). This case-control study suggests that the odds of firework-related ocular trauma were slightly higher among residents of areas where fireworks were legal compared with residents of areas where fireworks were banned. Although these results suggest that local firework bans may be associated with a small reduction in the odds of firework-related ocular trauma, additional studies are warranted to assess what actions might lead to greater reductions.


Case Report: Successful Lung Transplantation from a Donor Seropositive for Trypanosoma cruzi Infection (Chagas Disease) to a Seronegative Recipient.
The increasing shortage of organs for transplantation has prompted transplant programs to investigate the use of extended criteria donors, such as those with transmissible infectious diseases. Successful cases of organ transplantation (mostly kidney and liver) from Trypanosoma cruzi seropositive donors to seronegative recipients have been reported. We present a case of lung transplantation from a donor serologically positive for Chagas disease to a seronegative recipient, and provide a review of the literature. Left single lung transplantation was performed in a 44-year-old Spanish woman presenting with interstitial lung disease in February 2016. The deceased donor was a Colombian immigrant living in Spain who was serologically positive for Chagas disease. Oral administration of 5 mg/kg/day benznidazole divided in three doses for 60 days was given for specific Chagas disease prophylaxis after transplantation. Periodic follow-up with serological reverse transcription polymerase chain reaction to detect T. cruzi DNA were performed until 6 months after the end of treatment. All results were negative, indicating that transmission of T. cruzi had not occurred. In a review of the literature, two similar cases were identified in Argentina and the United States. In both cases T. cruzi infection was detected posttransplant in the recipients, after which they were treated with benznidazole. The course of the patient described herein confirms that lungs from donors with chronic T. cruzi infection can be used successfully as allografts, and that posttransplant prophylaxis with benznidazole may reduce the probability of transmission of T. cruzi to the recipient.


Antiinflammatory Activity of a Novel Folic Acid Targeted Conjugate of the mTOR Inhibitor Everolimus.
Folate receptor (FR)-β has been identified as a promising target for antimacrophage and antiinflammatory therapies. In the present study, we investigated EC0565, a folic acid-derivative of everolimus, as a FR-specific inhibitor of the mammalian target of rapamycin (mTOR). Because of its amphiphilic nature, EC0565 was first evaluated for water solubility, critical micelle formation, stability in culture and FR-binding specificity. Using FR-expressing macrophages, the effect of EC0565 on mTOR signaling and cellular proliferation was studied. The pharmacokinetics, metabolism and bioavailability of EC0565 were studied in normal rats. The in vivo activity of EC0565 was assessed in rats with adjuvant arthritis, a "macrophage-rich" model with close resemblance to rheumatoid arthritis. EC0565 forms micellar aggregates in physiological buffers and demonstrates good water solubility as well as strong multivalent FR-binding capacity. EC0565 inhibited mTOR signaling in rat macrophages at nanomolar concentrations and induced G0/G1 cell cycle arrest in serum-starved RAW264.7 cells. Subcutaneously administered EC0565 in rats displayed good bioavailability and a relatively long half-life (~12 h). When given at 250 nmol/kg, EC0565 selectively inhibited proliferating cell nuclear antigen expression in thioglycollate-stimulated rat peritoneal cells. With limited dosing regimens, the antiarthritic activity of EC0565 was found superior to that of etanercept, everolimus and a nontargeted everolimus analog. The in vivo activity of EC0565 was also comparable to that of a folate-targeted aminopterin. Folate-targeted mTOR inhibition may be an effective way of suppressing activated macrophages in sites of inflammation, especially in nutrient-deprived conditions, such as in the arthritic joints. Further investigation and improvement upon the physical and biochemical properties of EC0565 are warranted.


Alcoholism and the forensic-psychiatric patient: a comparative study.
A group of male forensic psychiatric patients was compared to groups of non-forensic psychiatric patients, hospitalized alcoholics and general hospital admissions on a modified version of the self-administered Michigan Alcoholism Screening Test. Results indicated more alcoholic related problems in both psychiatric groups than the general hospital group, but less than for the hospitalized alcoholics. The scores of the two psychiatric groups, however, did not differ significantly. It was concluded that alcoholism was non-additively associated with both criminality and psychiatric disturbance. It was suggested that the degree of alcohol related problems in psychiatric populations warrants careful screening and treatment even when alcoholism is not reflected in the psychiatric diagnosis.


Update on the molecular pathogenesis and clinical treatment of mantle cell lymphoma: report of the 10th annual conference of the European Mantle Cell Lymphoma Network.
Mantle cell lymphoma (MCL) is a distinct subtype of malignant lymphoma characterized by the chromosomal translocation t(11;14)(q13;q32), resulting in constitutional overexpression of cyclin D1 and cell cycle dysregulation in virtually all cases. Clinically, MCL shows an aggressive clinical course with a continuous relapse pattern and a median survival of only 3-5 years. However, recently a subset of up to 15% long-term survivors has been identified with a rather indolent clinical course. Advanced stage disease is usually apparent already at first clinical manifestation; in general, conventional chemotherapy is only palliative and median duration of remissions is only 1-2 years. In 2000, the European MCL Network ( http://www.european-mcl.net ) was founded, which consists of 15 national lymphoma study groups supplemented by experts in hematopathology, cytogenetics and molecular genetics. During the last decade, the European consortium has successfully initiated the largest phase III trials in MCL worldwide. In the current study generation, the addition of high-dose Ara-C to a rituximab, cyclophosphamide, doxorubicin, vincristine and prednisone (R-CHOP)-like regimen followed by myeloablative consolidation achieved a significant improvement of progression-free survival. Similarly, in elderly patients, rituximab maintenance until progression led to a marked prolongation of remission duration. Emerging strategies include proteasome inhibitors, immune modulatory drugs (IMiDs), mammalian target of rapamycin (mTOR) inhibitors and others, all based on the dysregulated control of cell cycle machinery and impairment of several apoptotic pathways. Combination strategies are currently being investigated in numerous trials, but their introduction into clinical practice and current treatment algorithms remains a challenge. Future strategies will apply individualized approaches according to the molecular risk profile of the patient. At the annual conference in Warsaw, recent results of molecular pathogenesis, analyses of current clinical trials and new study concepts were discussed.


Asialoagalacto-human chorionic gonadotropin, a carbohydrate-modified variant of human chorionic gonadotropin, antagonizes the stimulatory actions of bovine thyroid-stimulating hormone on thyroid function and HLA-DR expression in human thyroid in vitro and in vivo.
The concept of using thyroid-stimulating hormone (TSH) receptor antagonists in the management of Graves' disease is intriguing. Therefore, we investigated a TSH receptor antagonist derived from human chorionic gonadotropin (hCG) with respect to TSH receptor binding, adenylate cyclase activity, thyroid hormone release, and HLA class II antigen expression in vitro and in an in vivo model. A variant of hCG, asialoagalacto-hCG, like asialo-hCG and unlike hCG itself, inhibited both 125I-bTSH binding and cAMP response to bTSH in human thyroid membranes. However, like intact or deglycosylated hCG and unlike asialo-hCG, asialoagalacto-hCG displayed a limited affinity for hepatic asialoglycoprotein receptors, a likely marker for its in vivo turnover rate. It proved capable of inhibiting bTSH-stimulated thyroid hormone release in human thyroid slices as well as in the nude mouse bearing human thyroid transplants. It also prevented bTSH induced hypertrophy of transplanted thyrocytes. Further, HLA-DR expression induced by bTSH in the presence of gamma-interferon on human thyrocytes was inhibited. In conclusion, we present evidence that asialogalacto-hCG antagonizes bTSH actions on thyroid function and HLA-DR expression in human thyroid in vitro and, more importantly, in an in vivo model. Hence, the hCG variant described here or similar agents should warrant further exploration in the study and treatment of Graves' disease.


Evaluation of biogas production from seaweed in batch tests and in UASB reactors combined with the removal of heavy metals.
Seaweed can be anaerobically digested for the production of energy-rich methane. However, the use of seaweed digestate as a fertilizer may be restricted because of the high heavy metal content especially cadmium. Reducing the concentration of heavy metals in the digestate will enable its use as a fertilizer. In this laboratory-scale study, the potential of seaweed and its leachate in the production of methane were evaluated in batch tests. The effect of removing the heavy metals from seaweed leachate was evaluated in both batch test and treatment in an upflow anaerobic sludge blanket (UASB) reactor. The heavy metals were removed from seaweed leachate using an imminodiacetic acid (IDA) polyacrylamide cryogel carrier. The methane yield obtained in the anaerobic digestion of seaweed was 0.12 N l CH(4)/g VS(added). The same methane yield was obtained when the seaweed leachate was used for methane production. The IDA-cryogel carrier was efficient in removing Cd(2+), Cu(2+), Ni(2+) and Zn(2+) ions from seaweed leachate. The removal of heavy metals in the seaweed leachate led to a decrease in the methane yield. The maximum sustainable organic loading rate (OLR) attained in the UASB reactor was 20.6 g tCOD/l/day corresponding to a hydraulic retention time (HRT) of 12 h and with a total COD removal efficiency of about 81%. Hydrolysis and treatment with IDA cryogel reduced the heavy metals content in the seaweed leachate before methane production. This study also demonstrated the suitability of the treatment of seaweed leachate in a UASB reactor.


Chinese calligraphic handwriting practice promotes positive affect in adolescents: Converging evidence from correlational and experimental designs.
Chinese calligraphic handwriting (CCH) practice has proven to be beneficial to several aspects of psychological health and has been considered a complementary therapy for mental disorders. However, less is known about whether CCH practice benefits an individual's positive affect, a core component of affective well-being. Therefore, we examined this question in Chinese adolescents by combining correlational and experimental methods. In Study 1 (N = 1460), we found that high school students who practiced CCH had higher positive affect in their daily lives than those who did not practice CCH. In Study 2, a training experiment was conducted with college students, who were randomly assigned to receive either CCH training or no treatment. A six-day short-term CCH training led to a significant benefit in positive affect in the training group (N = 23) relative to the control group (N = 27). Together, our study provides converging evidence that CCH practice can promote positive affect in adolescents, further supporting the promotion of CCH in mental health practice.


Are integrins involved in the aggregatory and phagocytic behaviour of fish haemostatic cells?
The involvement of a putative integrin-like fibrinogen receptor in the aggregatory and phagocytic behaviour of thrombocytes (platelet equivalents of fish) from the rainbow trout Oncorhynchus mykiss was studied. Aggregation of trout thrombocytes was induced by the thromboxane mimetic U-46619 in the presence of trout fibrinogen. Thrombocyte aggregation was inhibited by the tetrapeptide RGDS, but not by RGES or fibrinogen binding inhibitor peptide (HHLGGAKQAGDV). A range of monoclonal antibodies against the human platelet integrin alphaIIbbeta3 (anti-CD41a, anti-beta3 and LK7r) showed no reactivity with trout thrombocytes. Subsequently, a panel of monoclonal antibodies was raised against thrombocyte membrane preparations in an attempt to obtain an antibody against the putative integrin fibrinogen receptor. Of these monoclonal antibodies, four were found to inhibit thrombocyte aggregation, namely 12G2, 30D8, 32F8 and 32H10. The antibody 32H10 was shown significantly to inhibit the attachment of thrombocytes to immobilised trout fibrinogen, suggesting that it and the other antibodies recognise the putative fibrinogen receptor on trout thrombocytes. FITC-labelled Bacillus cereus were employed as test particles to prove that thrombocytes internalise bacteria via an active process and not simply by passive sequestration into the open canalicular system. Preincubation of bacteria with trout fibrinogen resulted in a significant increase in the number of thrombocytes exhibiting phagocytosis. This enhancement of phagocytosis by preincubation of B. cereus with trout fibrinogen could be inhibited by the tetrapeptide RGDS, but not by RGES, hence implicating the putative fibrinogen receptor in the internalisation of microorganisms. The relevance of these findings to the possible existence of an integrin-like receptor on trout thrombocytes is discussed.


A case for a joint strategy of diversified bet hedging and plasticity in the pea aphid wing polyphenism.
Phenotypic plasticity and diversified bet hedging are strategies for coping with variable environments. Plasticity is favoured when an organism can predict future conditions using environmental cues, while bet hedging is favoured when predictive cues are not available. Theoretical analyses suggest that many organisms should use a mixture of both strategies, because environments often present both scenarios. Here, we examine if the pea aphid wing polyphenism, a well-known case of plasticity, is potentially a mixture of plasticity and bet hedging. In this polyphenism, asexual females produce more winged offspring in crowded conditions, and wingless offspring in uncrowded conditions. We find that pea aphids use plasticity to respond to crowding and we find considerable genetic variation for this response. We further show that individual aphids produce both winged and wingless offspring, consistent with the variability expected in a bet hedging trait. We conclude that the pea aphid wing polyphenism system is probably a mixture of plasticity and bet hedging. Our study adds to a limited list of empirical studies examining mixed strategy usage, and suggests that mixed strategies may be common in dispersal traits.


Psychiatric investigation of 18 bereaved pet owners.
This survey explores the grief associated with the loss of a pet, and was carried out using a self-administered questionnaire. The questionnaires were handed out to 50 bereaved pet owners attending a public animal cremation service, and we received 18 responses. Participants responded within 0 to 44 (median 4) days of the death of their pet. Although most mental health problems immediately following mourning are presumed to be normal grief reactions, on the basis of several psychiatric scales, 8 of the 16 valid responses indicated depression and/or neurosis. Statistical analyses showed that the following factors were significantly associated with grief reactions: age of owner, other stressful life events, family size, age of deceased animal, rearing place, and preliminary veterinary consultation.


Body image and self-esteem in disorders of sex development: A European multicenter study.
Disorders/differences of sex development (DSD) refer to congenital conditions with atypical sex development and are associated with psychosexual issues. The aim of this study was to assess body image and self-esteem across the DSD spectrum and to study the impact of diagnosis and mediating characteristics. Data collection was part of dsd-LIFE, a cross-sectional study conducted by 14 European expert clinics on wellbeing and health care evaluation of adults diagnosed with DSD. Main outcome measures in the present analyses were the Body Image Scale and Rosenberg Self-Esteem Scale. Additional data were obtained on treatments, openness, body embarrassment, sexual satisfaction, anxiety, and depression. The participating sample (n = 1,040) included 226 classified as Congenital Adrenal Hyperplasia, 225 as Klinefelter Syndrome, 322 as Turner Syndrome, and 267 as conditions with 46,XY karyotype. Many participants had received hormonal and surgical treatments. Participants scored lower on body image and self-esteem compared to control values, whereas each diagnosis showed different areas of concern. Limited openness, body embarrassment, and sexual issues were frequently reported. Overall body satisfaction was associated with BMI, hormone use, openness, body embarrassment, anxiety, and depression; genital satisfaction with age at diagnosis, openness, sexual satisfaction, and body embarrassment. Body embarrassment, anxiety, and depression predicted lower self-esteem. While each DSD showed specific issues related to body image and self-esteem, our findings indicate that the related factors were similar across the conditions. Clinical care on this subject could be improved by giving specific attention to factors like openness, body embarrassment, sexuality, anxiety, and depression. (PsycINFO Database Record


Ethanol increases the activity of Ca(++)-dependent K+ (mslo) channels: functional interaction with cytosolic Ca++.
Ethanol (EtOH) reversibly activates large conductance, Ca(++)-activated K+ (BK) channels in rat neurohypophysial terminals, an effect that probably contributes to the inhibition of vasopressin release by this drug. Heterogeneity in the terminal channel population makes it difficult to determine the mechanisms underlying this activation. Here, we report the effects of EtOH on the steady-state activity of BK channels cloned from mouse brain (mslo, alpha subunit) and expressed in Xenopus oocytes. EtOH reversibly increased mslo channel activity in excised patches, showing a potency (EC50 = 24 mM) similar to that reported using native channels. EtOH activation was observed under conditions that make it highly improbable that it is mediated by freely diffusible second messengers, or secondary to G-protein modulation. Rather, it probably results from a functional interaction between the drug and the channel alpha subunit. Activation occurred without increase in the number of functional channels present in the patch and resulted from actions that were a function of EtOH concentration: at < or = 10 mM, activation was due to a decrease in the channel mean closed time, whereas between 25 and 100 mM, activation was due to both a decrease in the mean closed time and an increase in the mean open time. The characteristic high unitary conductance and ionic selectivity of BK channels were unaltered by the drug. Whereas the voltage dependence of channel gating remained unchanged, channel activation mediated by the response of the Ca(++)-sensing site(s) to increases in the concentration of intracellular calcium, [Ca++]ic, was reduced by EtOH. This finding is consistent with EtOH and [Ca++]ic behaving functionally as partial and full agonists of mslo channels, respectively. Because the potentiation of mslo activity by the drug decreased as Ca++ levels were increased, EtOH-activation of BK channels would be most evident when [Ca++]ic is near resting levels, rather than during periods of high activity and Ca++ influx.


[Heart failure: critical patients].
Patients with heart failure in the "real world" are often elderly and with multiple comorbid diseases. These conditions create a dilemma for the physician responsible for the treatment of heart failure and are associated with a substantial underutilization of evidence-based treatments. Clarifying the prognostic impact of comorbidities in heart failure could provide more precise risk stratification and optimize the management of these patients. The negative prognostic impact of concomitant diseases has been shown in several studies: in the TEMISTOCLE study, carried out in Italy on patients hospitalized for heart failure in Internal Medicine and Cardiology wards, the presence of comorbidities was associated with higher in-hospital mortality and prolonged length of stay. In the IN-CHF registry, enrolling out patients with heart failure in a cardiological setting, the rate of coexisting diseases is not very high according to the epidemiological characteristics of this population. Renal impairment, particularly in patients >70 years old, and chronic obstructive pulmonary disease (COPD) are frequent comorbid diseases in heart failure. Renal impairment has been recognized as an independent risk factor for morbidity and mortality in heart failure while the role of COPD is controversial. Patients with renal dysfunction and COPD have largely been excluded from randomized controlled trials for safety reasons, so data are scarce. In the IN-CHF registry the prevalence of elderly patients with renal impairment (serum creatinine > or = 2 mg/dl and age > or = 70 years) is 5.1%; this subgroup of patients has an increased risk for both 1-year death (28.1 vs 11.2%) and hospital admission (34.9 vs 22.5%) compared with the remaining population. The prescription pattern has been evaluated in the last years (2003-2005) and shows that angiotensin system inhibitors (angiotensin-converting enzyme inhibitors and angiotensin receptor blockers) as well as beta-blockers are less prescribed in these patients (78.9 vs 86.1% and 42.2 vs 55.9%, respectively). The prevalence of patients with COPD in the registry was 13.2%: considerable differences in COPD prevalence estimates exist in the general population depending on many factors such as method for diagnosis or lack of agreement on diagnostic criteria. COPD patients were older and with more severe symptoms; with respect to the pharmacological treatment, beta-blockers are significantly less prescribed in COPD patients while a similar proportion of patients are receiving angiotensin system antagonists. The adjusted analysis shows that COPD in not an independent predictor of 1-year mortality in this population while it is independently associated with 1-year all-cause hospitalization. Non-cardiovascular hospital admissions seem to be more influenced by the presence of this comorbidity than cardiovascular admissions.


Cortical microstructural associations with CSF amyloid and pTau.
Diffusion MRI (dMRI) can be used to probe microstructural properties of brain tissue and holds great promise as a means to non-invasively map Alzheimer's disease (AD) pathology. Few studies have evaluated multi-shell dMRI models such as neurite orientation dispersion and density imaging (NODDI) and mean apparent propagator (MAP)-MRI in cortical gray matter where many of the earliest histopathological changes occur in AD. Here, we investigated the relationship between CSF pTau181 and Aβ1-42 burden and regional cortical NODDI and MAP-MRI indices in 46 cognitively unimpaired individuals, 18 with mild cognitive impairment, and two with dementia (mean age: 71.8 ± 6.2 years) from the Alzheimer's Disease Neuroimaging Initiative. We compared findings to more conventional cortical thickness measures. Lower CSF Aβ1-42 and higher pTau181 were associated with cortical dMRI measures reflecting less hindered or restricted diffusion and greater diffusivity. Cortical dMRI measures, but not cortical thickness measures, were more widely associated with Aβ1-42 than pTau181 and better distinguished Aβ+ from Aβ- participants than pTau+ from pTau- participants. dMRI associations mediated the relationship between CSF markers and delayed logical memory performance, commonly impaired in early AD. dMRI metrics sensitive to early AD pathogenesis and microstructural damage may be better measures of subtle neurodegeneration in comparison to standard cortical thickness and help to elucidate mechanisms underlying cognitive decline.


Movement Disorder-Childhood Rating Scale: A Sensitive Tool to Evaluate Movement Disorders.
The Movement Disorder-Childhood Rating Scale represents a new tool for assessment of movement disorders during developmental age. In this study, we evaluated a cohort of 68 patients affected by various types of movement disorders and treated with specific drugs over one year to verify the usefulness of the Movement Disorder-Childhood Rating Scale. The participants were divided into two groups according to their ages (0-3 years; 4-18 years) and were evaluated using Movement Disorder-Childhood Rating Scale 0-3 or 4-18 at baseline (i.e., before starting pharmacological treatment [T0], after 6 months [T1], and after 12 months [T2] of treatment. Univariate repeated measures analysis of variance with a Greenhouse-Geisser correction by SPSS 20 was performed to analyze the scale responsiveness for the three indices (e.g., Index I, Index II, Global Index) in each group with time (T0, T1, and T2). In addition, the Bonferroni test was performed to identify the source of significant differences among means. Significant differences were found between time points (T1 versus T0, T2 versus T0, and T2 versus T1) in both scales for all indexes with the exception for T2 versus T1 for Index II in both scales and for T2 versus T1 for the Global Index in the older age group. There was no significant correlation between observed changes in the scores and the age of the children, either for Movement Disorder-Childhood Rating Scale 0-3 or 4-18. Our results suggest that Movement Disorder-Childhood Rating Scale is a suitable tool to detect changes independently from age and could be used as outcome measure for clinical trials.


Severe infective keratitis leading to hospital admission in New Zealand.
To identify key risk factors and the management and outcome of severe infective keratitis leading to public hospital admission in New Zealand. Over a 2 year period, all admissions of presumed infective keratitis to Auckland Hospital were identified. The clinical records of all 103 cases were retrospectively reviewed with respect to clinical features, risk factors, management, and outcomes. The mean time from first symptoms or signs and presentation to hospital was 8.9 (SD 15.5) days. The majority of subjects, 88%, had at least one of the risk factors commonly associated with infective keratitis including previous ocular surgery (30%), contact lens wear (26%), topical corticosteroid use (25%), and ocular trauma (24%). Corneal scraping was performed in 92% and of a total of 105 scrapes, 71% were positive. Bacteria were isolated in all these cases, the majority being Gram positive organisms (72%). The most common isolates identified were coagulase negative Staphylococcus (16%), Propionibacterium acnes (14%), Staphylococcus epidermidis (11%), and Streptococcus pneumoniae (9%). In addition, yeasts were isolated in 5%, fungi in 4%, virus in 2%, and chlamydia in 1%. Importantly, polymicrobial infection accounted for 33% of culture positive cases. Antimicrobial treatment was changed on the basis of culture results in 17 cases (16.5%). Median initial visual and final best corrected visual acuity was 6/36-6/48 (logMAR 0.86) (IQR 0.39-2.00) and 6/12-6/15 (logMAR 0.360) (IQR 0.15-1.70), respectively. Previous ocular surgery and topical corticosteroid use were significantly associated with poorer visual acuity. The mean hospital stay was 5.8 days and the median 4.0 (IQR 2.0-8.0) days. Longer duration of stay was associated with the presence of hypopyon, larger ulcers, previous ocular surgery, and poor visual acuity. Infectious keratitis is an important cause of ocular morbidity. A significant proportion of cases have potentially modifiable risk factors. Previous ocular surgery and topical corticosteroid use, in particular, were associated with poorer visual outcomes. Many cases of severe keratitis might be avoided, or their severity reduced, by appropriate education of patients and ophthalmologists.


The boarding experience from the patient perspective: the wait.
We sought to better understand the experience of being a boarder patient. We conducted a qualitative study between March and August 2012 to examine the experience of boarding in an urban, teaching hospital emergency department (ED). We included boarder patients and selected patients based on a convenience sample. Interviews were semistructured, consisting of eight main open-ended questions. Interviews were transcribed; codes were generated and then organised into categorises and subsequently into one theme. We concluded analysis when we achieved thematic saturation. Our institutional review board approved this study. Our final sample included 18 patients. The average age was 62.3 years. Patients characterised waiting as central to their experience as a boarder patient. One patient stated, "Well if you have to wait for a bed you have to wait for a bed, it's terrible." Three categories exemplified this waiting experience: (1) there was often lack of communication; (2) patients experienced frustration during this waiting period; and yet (3) patients often differentiated the experience of waiting from the care they were receiving. Being a boarder patient was characterised as a waiting experience associated with poor communication and frustration. However, patients may still differentiate their feelings towards the wait from those towards the medical care they are receiving. Our data add more reason to eradicate the practice of ED boarding.


Quantitative evaluation of active thermography using contrast-to-noise ratio.
Active thermography is an infrared-based technique for nondestructive testing of materials. It often uses advanced evaluation techniques based on temperature spatial and temporal changes. Results of active thermography are contrast differences, which indicate possible defects in an inspected material. These differences cannot be quantified by temperature. This contribution is focused on the active thermography results' evaluation parameters and the contrast-to-noise ratio method, which can be used for quantitative evaluation of the results. Different result interpretation procedures are introduced. The influence of a selection method for indication and reference regions and the effect of image scaling on inspection results are discussed.


Radiological and survival comparison of four methods of fixation of a proximal femoral stem.
We compared the radiological appearances and survival of four methods of fixation of a femoral stem in 538 hips after follow-up for five or ten years. The fixation groups were: 1) press-fit shot-blasted smooth Ti-A1-V stem; 2) press-fit shot-blasted proximally ridged stem; 3) proximal hydroxyapatite (HA) coating; and 4) cementing. Survival analysis at five to ten years showed better results in the HA-coated (100% at five to six years) and cemented stems (100% at 5 to 6 years) than in the two press-fit groups. There was a higher mean rate of migration in the smooth and ridged Ti-A1-V shot-blasted press-fit groups (0.8 mm/year and 0.6 mm/year, respectively) when compared with the HA-coated and cemented prostheses (both 0.3 mm/year). More radiolucent lines and osteolytic lesions were seen in the press-fit groups than in either the HA-coated or cemented implants, with a trend for a lower incidence of both in the HA compared with the cemented group. Proximal osteopenia increased in the press-fit and cemented prostheses with time, but did not do so in the HA group. There was a higher incidence of resorption of the femoral neck with time in the cemented group than in the other three. We conclude that the HA and the cemented interfaces both provide secure fixation with a trend in favour of HA. The cemented prosthesis meets the suggested National Institutes of Health definition of 'efficacious' at ten years.


Central hyperexcitability as measured with nociceptive flexor reflex threshold in chronic musculoskeletal pain: a systematic review.
Chronic musculoskeletal conditions are increasingly conceived as involving altered central nervous system processing, and impaired nociceptive flexor reflex (NFR) appears to reflect altered central nervous system processing. The primary objective was to synthesize the evidence for impaired NFR in these conditions. The secondary objective was to evaluate the NFR stimuli parameters employed by reviewed studies. Electronic databases: MEDLINE, CINAHL, Embase, PEDro, Google Scholar, and Cochrane library were searched from the mid-1960s to June 2010. Experimental reports were systematically reviewed and meta-analysis (where possible) was performed. NFR thresholds and parameters of NFR stimuli were extracted. Sixteen trials were identified, 11 of which were suitable for inclusion in the meta-analysis. Compared to healthy controls, standardized mean differences in NFR threshold were significantly lower in subjects with primary headache (-0.45; 95% confidence interval [CI] -0.77 to -0.13, P=0.005), fibromyalgia (-0.63; 95% CI -0.93 to -0.34, P<0.0001), knee pain (-1.51; 95% CI -2.10 to -0.93, P<0.00001) and whiplash (-0.73; 95% CI -1.11 to -0.35, P=0.0002). Employed stimuli parameters vary between studies, with inter-pulse duration (P=0.044) being identified by multiple regression analysis as independent predictors of the variability in NFR threshold in healthy controls. The results indicate that there is evidence of central hyperexcitability in people with chronic musculoskeletal pain. Our review also suggests that shorter inter-pulse duration tends to yield smaller variability in NFR threshold. However, further research investigating optimal stimulation parameters is still warranted.


A simple, high-throughput method of protein and label removal from extracellular vesicle samples.
Evidence continues to increase of the clinical utility extracellular vesicles (EVs) as translational biomarkers. While a wide variety of EV isolation and purification methods have been implemented, few techniques are high-throughput and scalable for removing excess fluorescent reagents (e.g. dyes, antibodies). EVs are too small to be recovered from routine cell-processing procedures, such as filtration or centrifugation. The lack of suitable methods for removing unbound labels, especially in optical assays, is a major roadblock to accurate EV phenotyping and utilization of EV assays in a translational or clinical setting. Therefore, we developed a method for using a multi-modal resin, referred to as EV-Clean, to remove unbound labels from EV samples, and we demonstrate improvement in flow cytometric EV analysis with the use of this EV-Clean method.


YAP/TAZ for cancer therapy: opportunities and challenges (review).
YAP (Yes-associated protein) and its paralog TAZ (transcriptional co-activator with PDZ-binding motif) are the main downstream effectors of the Hippo signaling pathway. This pathway is an evolutionally conserved signal cascade, which plays pivotal roles in organ size control and tumorigenesis from Drosophila to mammals. Functionally, when the Hippo pathway is activated, YAP and TAZ will be sequestered in the cytoplasm and degraded. Conversely, when the Hippo pathway is deactivated, YAP and TAZ will translocate into nucleus and promote transcription of downstream genes by forming complexes with transcription factors, such as transcriptional enhancer factors (TEF; also referred to as TEAD), runt-domain transcription factors (Runx) and others. Most of these transcription factors belong to growth promoting or apoptosis-inhibition genes. It has been reported that the deactivation of the Hippo pathway, as well as up-regulation of YAP and TAZ was observed in many human cancers with a high frequency, which suggests that the Hippo pathway may be a potent target for developing anticancer drugs. In this review, we provide an overview of the Hippo pathway and summarize recent advances with respect to the role of YAP and TAZ in Hippo signaling pathway and cancer development. Furthermore, we describe the opportunities and challenges for exploit YAP and TAZ as potential therapeutic targets in cancer.


Tobacco, nicotine and harm reduction.
Tobacco smoking, sustained by nicotine dependence, is a chronic relapsing disorder, which in many cases results in lifelong cigarette use and consequent death of one out of two lifelong smokers from a disease caused by their smoking. Most toxicity due to cigarette smoking is related to the burning process. Models of harm reduction applied to tobacco suggest that use of non-combustible, less toxic, nicotine-containing products as a substitute for cigarette smoking would reduce the death toll arising from tobacco use. Available options include medicinal nicotine and smokeless tobacco products. The potential role of nicotine replacement therapy (NRT) products in a harm reduction strategy is currently severely restricted by strict regulations on dose, safety and potential addictiveness. As a result, NRT products are designed to provide much less nicotine, and deliver it to the brain more slowly, than cigarettes, which are widely accessible and poorly regulated. Smokeless tobacco (snus) has proved to be an acceptable reduced hazard alternative to smoking in Sweden, but supply of snus is illegal elsewhere in the European Union. To increase accessibility and reach more smokers, barriers to the use of NRT use need to be removed and more effective NRTs need urgently to be developed. Smokeless tobacco could also play an important role in harm reduction, but current European Union regulations and concerns over exploitation by tobacco companies currently preclude wider use. To improve public health there is an urgent need for an appropriate regulatory framework and regulatory authority at the European level, controlling both tobacco and nicotine products to ensure that the least harmful products are the most accessible.


Optimization of Critical Parameters for Carbodiimide Mediated Production of Highly Modified Chitosan.
An optimization of the 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and hydroxy benzotriazole mediated conjugation of the polysaccharide chitosan with functional carboxylic acids was shown. Optimal parameters that enable resource-efficient synthesis of highly functionalized chitosan were identified. In particular, use of only catalytic instead of stoichiometric amounts of hydroxy benzotriazole and tight control of pH in reaction mixture resulted in highly efficient incorporation of the desired moieties as side chains in chitosan. As a result, the model reactant 4-azidobenzoic acid was incorporated resulting in a degree of substitution of over 30% with very high coupling efficacy of up to 90%. Similar results were obtained with other carboxylic acids such as methacrylic acid, 3-(2-furyl) propionic acid and 3-maleimido propionic acid, highlighting the broad applicability of our findings for the functionalization of chitosan.


Technical and clinical description of a case of extensive anogenital Paget's disease associated with anal cancer treated by tomotherapy.
In this paper we describe a case of extramammary Paget's disease associated with anal cancer, which was successfully treated by intensity-modulated radiotherapy using tomotherapy with a simultaneous integrated boost and daily image guidance. The main pitfall in this report is the relatively short follow-up (1 year), which means that the evaluated data is promising but not conclusive. Considering the rarity and wide extension of our patient's Paget's disease in the anogenital region, and the lack of literature reports about curative radiotherapy in this particular setting, this case report may be considered the first related to extensive extramammary Paget's disease treated by tomotherapy.


[Plasma apolipoproteins B and AI in healthy individuals and patients with coronary atherosclerosis in a male population from Lithuania].
The paper discusses the results from a study into apolipoproteins B (apo-B) and AI (apo-AI) and apoB/apo-AI ratios in the blood of 188 healthy donors and 122 male patients with coronary atherosclerosis. Atherosclerotic lesions were coronary angiographically documented. The study was performed (in two age groups of patients: 1) those under 45 years and 2) those over 45 years. A relationship was examined between the levels of apolipoproteins and the lesion of one, two, three or more coronary arteries in the two age groups. The studies indicate that there are various mechanisms responsible for the pathogenesis of atherosclerosis in the age groups in question. It is concluded that the younger a patient is, the higher probability that elevated blood apo-B levels are essential in the pathogenesis of coronary atherosclerosis is. Determination of plasma apo-B and apo-AI levels and apo-B/apo-AI ratios enables a potential number of diseased coronary arteries to be predicted.


Impact of high glucose on metastasis of colon cancer cells.
To investigate the possible mechanism of how glucose promotes invasion and metastasis of colon cancer cells. CT-26 rat colorectal cancer cells were cultured in different concentrations of glucose environments (10, 20, and 30 mmol/L). Wound healing assay and transwell chamber invasion assay were utilized to test the migration and invasion, respectively. In order to understand the role of signal transducer and activator of transcription 3 (STAT3) in the process, STAT3 inhibitors, including Stattic (an STAT3 specific inhibitor) and small interfering RNA targeting STAT3, were used to block STAT3 function to evaluate their impact on CT-26 cell motion. To verify whether STAT3 and matrix metalloproteinase-9 (MMP-9) protein expression is associated with glucose-induced cell movement, Western blot was used to compare the differences in the expression of MMP-9 and STAT3 in cells incubated with and without STAT3 inhibitors in high glucose condition. In both wound healing and invasion assays, the migration and invasion of CT-26 cells increased gradually with the increase in glucose concentration. However, the glucose-induced migration and invasion were obviously inhibited by STAT3 inhibitors (P<0.05). Similarly, in Western blot assessment, both MMP-9 and STAT3 expression increased under a high glucose environment and the highest expression was achieved when 30 mmol/L glucose was used. However, in cells treated with 30 mmol/L mannitol, either MMP-9 or STAT3 expression did not increase (P>0.05). When STAT3 inhibitors were added in the 30 mM glucose group, not only STAT3 but also MMP-9 expression decreased significantly (P<0.05). Our study provides evidence that glucose can promote both migration and invasion of CT-26 cells, and that the STAT3-induced MMP-9 signal pathway is involved in this process.


[Adaptation experiences in South Korea of men defecting from North Korea].
The study was done to explore meanings and essence of the experience in South Korea of men defectors from North Korea. Data were collected from March, 2011 to May, 2012, through in-depth interviews with ten men defectors. Data analysis was conducted using the process of hermeneutic phenomenological reflection. Eight essential themes were extracted; 'buoyant expectation for a new life', 'feeling guilty about family left behind in North Korea', 'inability to become acclimatized due to communication difficulties', 'inability to socialize with South Koreans due to different lifestyles', 'finding strength through trustworthy acquaintances', 'continuing reconciliation with oneself while trying to assimilate into South Korean culture', 'self-realization of one's original self', and 'continuing to feel out the possibility of a future in South Korea'. The findings indicate that North Korean men who defect to South Korea shape their identity through three phases: forming self-image during escape from North Korea, trying to become accustomed to South Korean society, and finding their own identity by self-realization of their original self. Eventually, the whole process enables them to recover their identity, to feel a sense of belonging, and to discover possibilities for a better future.


Standardized inspections of food premises during the 2004 Athens Olympic Games: descriptive analysis and risk factors for unsatisfactory results.
Standardized inspections of food premises are part of environmental health systems implemented worldwide. The food safety strategy for the 2004 Olympic Games included standardized inspections to ensure uniformity and consistency of procedures and effective electronic management of data. Inspections were carried out by 196 inspectors in the five Olympic cities: Athens, Thessalonica, Volos, Iraklio, and Patra. From January 2003 to September 2004, a total of 1,249 food premises were inspected. An unsatisfactory inspection result (C grade) was received by 347 (27.8%) food premises, a relatively satisfactory result (B grade) was received by 332 (26.6%), and a satisfactory result (A grade) was received by 570 (45.6%). About 16% of inspected premises did not hold a valid permit. Unsatisfactory inspection results were more frequent for premises located in the two largest Greek cities in comparison with the other smaller cities (relative risk = 1.95, 95% confidence interval [CI] = 1.36 to 2.80). Based on logistic regression analysis, unsatisfactory inspection results were positively associated with food premises that were not located on a ground floor (odds ratio [OR] = 2.56, 95% CI = 1.39 to 4.73) and negatively associated with application of hazard analysis critical control point (HACCP) principles (OR = 0.27, 95% CI = 0.10 to 0.71). Food hygiene education through formal training programs should be encouraged to improve compliance of food premises. Food premises located on hotel floors and serving buffet meals are at higher risk for unsatisfactory conditions. Businesses that implemented a HACCP system within their operations to ensure food safety operated under more hygienic conditions. Future inspections by Public Health Authorities should involve elements of audit after the legislation for the application of HACCP principles.


Hepatitis C virus genotypes in chronic hepatitis C of children.
Several hepatitis C virus (HCV) genotypes have been recently identified and genotype 1b has been correlated with severe liver disease and a poor response to interferon therapy. HCV infection in children is an interesting model for evaluation of the relationship between HCV genotypes and liver disease, because of its relatively short duration and the infrequent association with confounding cofactors. We have investigated HCV genotypes, using a dot-blot hybridization assay with genotype-specific probes, in 36 Italian children with chronic hepatitis C who were otherwise well and had no other underlying disease. Only four patients were symptomatic; liver histology, obtained in 33 patients, showed minimal hepatitis in 17 and mild chronic hepatitis in 16. Infection with HCV genotype 1b was found in 55.5% of patients, with a peak prevalence of 83% in children from southern Italy (P < 0.05 vs other regions). The remaining children were infected with HCV genotype 1a (16.6%), genotype 2 (11.1%), and mixed (10.9%) or undetermined (2.7%) genotypes. In one patient, HCV viraemia was never detected. There was no statistically significant correlation between genotype and age, sex, source of infection, alanine aminotransferase pattern and histological activity index. These results indicate that genotype 1 b is widespread among Italian children with chronic hepatitis C, although with significant geographical variations. It is not associated with a more severe liver disease, therefore suggesting that the greater severity of liver disease recently reported in adults could reflect the cumulative effects of disease duration and of interfering cofactors.


Molecular phylogeny of sequenced saccharomycetes reveals polyphyly of the alternative yeast codon usage.
The universal genetic code defines the translation of nucleotide triplets, called codons, into amino acids. In many Saccharomycetes a unique alteration of this code affects the translation of the CUG codon, which is normally translated as leucine. Most of the species encoding CUG alternatively as serine belong to the Candida genus and were grouped into a so-called CTG clade. However, the "Candida genus" is not a monophyletic group and several Candida species are known to use the standard CUG translation. The codon identity could have been changed in a single branch, the ancestor of the Candida, or to several branches independently leading to a polyphyletic alternative yeast codon usage (AYCU). In order to resolve the monophyly or polyphyly of the AYCU, we performed a phylogenomics analysis of 26 motor and cytoskeletal proteins from 60 sequenced yeast species. By investigating the CUG codon positions with respect to sequence conservation at the respective alignment positions we were able to unambiguously assign the standard code or AYCU. Quantitative analysis of the highly conserved leucine and serine alignment positions showed, that 61.1% and 17% of the CUG codons coding for leucine and serine, respectively, are at highly conserved positions, while only 0.6% and 2.3% of the CUG codons, respectively, are at positions conserved in the respective other amino acid. Plotting the codon usage onto the phylogenetic tree revealed the polyphyly of the AYCU with Pachysolen tannophilus and the CTG clade branching independently within a time span of 30 to 100 Mya.


Map making in the 21st century: charting breast cancer susceptibility pathways in rodent models.
Genetic factors play an important role in determining risk and resistance to increased breast cancer. Recent technological advances have made it possible to analyze hundreds of thousands of single nucleotide polymorphisms in large-scale association studies in humans and have resulted in identification of alleles in over 20 genes that influence breast cancer risk. Despite these advances, the challenge remains in identifying what the functional polymorphisms are that confer the increased risk, and how these genetic variants interact with each other and with environmental factors. In rodents, the incidence of mammary tumors varies among strains, such that they can provide alternate ideas for candidate pathways involved in humans. Mapping studies in animals have unearthed numerous loci for breast cancer susceptibility that have been validated in human populations. In a reciprocal manner, knockin and knockout mice have been used to validate the tumorigenicity of risk alleles found in population studies. Rodent studies also underscore the complexity of interactions among alleles. The fact that genes affecting risk and resistance to mammary tumors in rodents depend greatly upon the carcinogenic challenge emphasizes the importance of gene x environment interactions. The challenge to rodent geneticists now is to capitalize on the ability to control the genetics and environment in rodent models of tumorigenesis to better understand the biology of breast cancer development, to identify those polymorphisms most relevant to human susceptibility and to identify compensatory pathways that can be targeted for improved prevention in women at highest risk of developing breast cancer.


EPIYA motif patterns among Cuban Helicobacter pylori CagA positive strains.
It is known that polymorphisms in C-terminal region of CagA influence gastric disease development on Helicobacter pylori infection. Additionally, the geographic distribution of these polymorphisms has been associated with the appearance of more severe gastroduodenal pathologies. Objective. To determine the CagA phosphorylation motifs pattern (EPIYA pattern) in Cuban H. pylori isolates, and to study its association with patient´s pathologies. DNAs from 95 H. pylori cagA-positive strains were used to amplify the 3´ variable region of cagA gene by PCR using two different strategies. Additionally, new primers were designed to identify either Western or Eastern CagAEPIYA motiftype by PCR. To confirm the PCR results, PCR products from 14 representative isolates were purified and sequenced. The distribution of the EPIYA motif found was: 2 AB (2.1 %), 1 AC (1.1 %), 1 BC (1.1 %), 70 ABC (73.6 %), 19 ABCC (20 %), and 2 ABCCC (2.1 %). Sequencing analysis confirmed the PCR classification in the 14 studied strains and showed three strains with unusual nucleotide sequences, not reported before. Distribution of the EPIYA-ABC pattern was equivalent in all pathologies (78.9 % in gastric ulcer, 72.5 % in duodenal ulcer and 72.2 % in non-ulcer dyspepsia). The PCR results using the new primers confirmed that all studied strains carried the Western CagA type. No specific EPIYA motif was associated with peptic ulcer. This is the first report that shows EPIYA motif distribution in H. pylori isolates from the Caribbean region.