Risk factors for treatment-related death in elderly patients with aggressive non-Hodgkin's lymphoma: results of a multivariate analysis.
It has been suggested that age is associated with chemotherapy-related death in patients with non-Hodgkin's lymphoma (NHL) treated with doxorubicin-containing chemotherapy. The purpose of this study was to evaluate the relative influence of increasing age and other clinical parameters on the occurrence of treatment-related death in elderly patients with intermediate- or high-grade NHL treated with cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP) chemotherapy. A retrospective study of patients 60 years of age or older with intermediate- or high-grade NHL treated with CHOP chemotherapy in a single cancer center. The following variables were recorded: age (60 to 69, 70 to 79, and 80 to 94 years), histology (Working Formulation [WF] D, E, F, G, and H), Ann Arbor stage, B symptoms, extranodal involvement, bulky disease (> 7 cm), performance status (Eastern Cooperative Oncology Group [ECOG] scale), International Prognostic Index (IPI score), serum lactate dehydrogenase (LDH) level and doxorubicin relative dose-intensity (RDI). The relationship between these features and treatment-related death was assessed in univariate and multivariate logistic regression analysis. From 1982 to 1991, 267 consecutive patients were treated. Median age was 70 years (range, 60 to 94 years). There were 35 toxic deaths. Sixty-three percent of the deaths occurred after the first cycle. Infection accounted for 82% of the toxic deaths. In the univariate analysis, the features associated with an increased risk of toxic death were ECOG performance status 2 to 4 (relative risk [RR], 7.82), B symptoms (RR, 3.38), diffuse large-cell histology (RR, 3.06), bulky disease (RR, 2.58), serum levels of LDH (RR, 2.53), and IPI score (RR, 2.46). The age groups did not show significance. In the regression model, performance status 2 to 4 was the only independent predictor of treatment-related death (RR, 3.52; 95% confidence interval [CI], 2.98 to 4.06). Our results show that in elderly patients with NHL treated with doxorubicin-based chemotherapy the risk for treatment-related death is associated with poor performance status rather than with increasing chronologic age.


Heterohelicenes with embedded P-chiral 1H-phosphindole or dibenzophosphole units: diastereoselective photochemical synthesis and structural characterization.
The oxidative photocyclization reactions of olefins that contain 1H-phosphindole or dibenzophosphole substituents have been applied to the synthesis of P/N-bi-heterosubstituted dimeric helicenes, as well as of new [6]- and [8]phosphahelicenes. In these photocyclization processes, the configuration of the stereogenic phosphorus center dictates the sense of helical chirality. Thus, by starting from enantiomerically pure P-menthylphosphole-oxide units, this method affords enantiopure helical compounds. The helical phosphine oxides were characterized by X-ray diffraction. After reduction of the phosphine-oxides, the corresponding helical phosphines have been used as ligands in transition-metal complexes. The X-ray crystal structure of a gold chloride complex of a [6]helicene is reported.


Slow and stored light by photo-isomerization induced transparency in dye doped chiral nematics.
Decelerating and stopping light is fundamental for optical processing, high performance sensor technologies and digital signal treatment, many of these applications relying on the ability of controlling the amplitude and phase of coherent light pulses. In this context, slow-light has been achieved by various methods, as coupling light into resonant media, Brillouin scattering in optical fibers, beam coupling in photorefractive and liquid crystal media or engineered dispersion in photonic crystals. Here, we present a different mechanism for slowing and storing light, which is based on photo-isomerization induced transparency of azo-dye molecules hosted in a chiral liquid crystal structure. Sharp spectral features of the medium absorption/dispersion, and the long population lifetime of the dye metastable state, enable the storage of light pulses with a significant retrieval after times much longer than the medium response time.


Even more extreme fertility insurance and the sex ratios of protozoan blood parasites.
Theory developed for malaria and other protozoan parasites predicts that the evolutionarily stable gametocyte sex ratio (z*; proportion of gametocytes that are male) should be related to the inbreeding rate (f) by the equation z*=(1-f)/2. Although this equation has been applied with some success, it has been suggested that in some cases a less female biased sex ratio can be favoured to ensure female gametes are fertilized. Such fertility insurance can arise in response to two factors: (i) low numbers of gametes produced per gametocyte and (ii) the gametes of only a limited number of gametocytes being able to interact. However, previous theoretical studies have considered the influence of these two forms of fertility insurance separately. We use a stochastic analytical model to address this problem, and examine the consequences of when these two types of fertility insurance are allowed to occur simultaneously. Our results show that interactions between the two types of fertility insurance reduce the extent of female bias predicted in the sex ratio, suggesting that fertility insurance may be more important than has previously been assumed.


The alpha-ketoisocaproate catabolism in human and rat livers.
Catabolism of alpha-ketoisocaproate in liver is mediated by cytosolic alpha-ketoisocaproate dioxygenase (KICD) and mitochondrial branched-chain alpha-keto acid dehydrogenase complex (BCKDC). The latter is believed to be involved in the main pathway of the KIC catabolism. In the present study, we measured the activities of KICD and BCKDC in human and rat livers. The KICD activity in human liver was 0.9 mU/g tissue, which was 14.2% of the total activity of BCKDC, and that in rat liver was 4.2 mU/g tissue, which was only 1.0% of the total activity, suggesting that KICD in human liver plays a relatively important role in the alpha-ketoisocaproate catabolism. The KICD activity in human liver was significantly increased by cirrhosis. In rat liver, the enzyme activity was markedly increased by physical training and streptozotocin-induced diabetes, but not by feeding of a diet rich in branched-chain amino acids, although BCKDC activity was increased by feeding of the diet.


Automated ligand- and structure-based protocol for in silico prediction of human serum albumin binding.
Plasma protein binding has a profound impact on the pharmacokinetic and pharmacodynamic properties of many drug candidates and is thus an integral component of drug discovery. Nevertheless, extant methods to examine small-molecule interactions with plasma protein have various limitations, thus creating a need for alternative methods. Herein we present a comprehensive and cross-validated in silico workflow for the prediction of small-molecule binding to Human Serum Albumin (HSA), the most ubiquitous plasma protein. This protocol reliably predicts small-molecule interactions with HSA, including a binding affinity calculation using multiple linear regression methods, binding site prediction using a naive-Bayes classifier, and a three-dimensional binding pose using induced fit docking. Furthermore, this workflow is implemented in a portable and automated format that can be downloaded and used by other end users, either as is or with customization.


UPF1 ATPase autoinhibition and activation modulate RNA binding kinetics and NMD efficiency.
The RNA helicase UPF1 interacts with mRNAs, mRNA decay machinery, and the terminating ribosome to promote nonsense-mediated mRNA decay (NMD). Structural and biochemical data have revealed that UPF1 exists in an enzymatically autoinhibited "closed" state. Upon binding the NMD protein UPF2, UPF1 undergoes an extensive conformational change into a more enzymatically active "open" state, which exhibits enhanced ATPase and helicase activity. However, mechanically deficient UPF1 mutants can support efficient NMD, bringing into question the roles of UPF1 enzymatic autoinhibition and activation in NMD. Here, we identify two additional important features of the activated open state: slower nucleic acid binding kinetics and enhanced ATP-stimulated nucleic acid dissociation kinetics. Computational modeling based on empirical measurements of UPF1, UPF2, and RNA interaction kinetics predicts that the majority of UPF1-RNA binding and dissociation events in cells occur independently of UPF2 binding. We find that UPF1 mutants with either reduced or accelerated dissociation from RNA have NMD defects, whereas UPF1 mutants that are more dependent on UPF2 for catalytic activity remain active on well-established NMD targets. These findings support a model in which the kinetics of UPF1-mRNA interactions are important determinants of cellular NMD efficiency.


Practice profile time survey and government policy.
A practice profile time survey developed by FDI/WHO JWG 9 was employed by Hong Kong Dental Association in collaboration with the Faculty of Dentistry, University of Hong Kong, as part of an exercise to assess oral health manpower requirements in Hong Kong. The results highlighted specific features of private dental practice reflecting needs and demands of the indigenous population. The protocol was found to be reliable and could be used by national associations to collect information required in the monitoring of oral health services, as well as formulation of government policies.


The immunoglobulin D Fc receptor expressed on fibroblast-like synoviocytes from patients with rheumatoid arthritis contributes to the cell activation.
Immunoglobulin IgD might play an important role in autoimmune diseases, but the function of IgD has remained elusive, despite multiple attempts to define its biological function. Fibroblast-like synoviocytes (FLSs) are specialized cells of the synovium that play a key role in the pathogenesis of rheumatoid arthritis (RA). In this study we explored the possible roles of excessive IgD expression on the function of FLSs from RA patients (RA-FLSs). We showed that IgD Fc receptor (IgDR) was constitutively expressed on FLSs, and was significantly elevated in RA-FLSs compared with FLSs prepared from synovial tissues of healthy controls (HC-FLSs). Furthermore, IgDR was mainly detected on the cell surface and in the cytoplasm. We further detected the intrinsic binding affinity of IgD to IgDR on HC-FLSs with an equilibrium dissociation constant (KD) of 0.067 nmol/L. Incubation of RA-FLSs with IgD (1-10 μg/mL) for 48 h dose-dependently promoted the expression of IgDR, and stimulated the production of inflammatory cytokines and chemokines, such as IL-1β, IL-6, monocyte chemotactic protein (MCP)-1, TNF-α and receptor activator of nuclear factor-κB ligand (RANKL), thus potentially contributing to IgD-IgDR crosslinking. Moreover, incubation with IgD (0.1-10 μg/mL) for 48 h dose-dependently enhanced viability for both HC-FLSs and RA-FLSs. Our results demonstrate that IgDR is expressed on RA-FLSs and contributes to the activation of FLSs, and suggest that IgD-IgDR is a potential novel immunotherapeutic target for the management of RA.


Engaging medical students in occupational and environmental medicine--a new approach.
For a number of reasons, engaging the interest of medical students in the discipline of occupational and environmental medicine (OEM) can be challenging. To renew a curriculum in OEM within a graduate medical programme with an emphasis on student involvement to maximize their interest in the topic. A second year student cohort of a 4 year graduate medical programme was surveyed as to their preferences for the content of a short course of OEM embedded in their medical course. The course was extensively rewritten as a result of the student survey, with a number of topics deleted from the old course and new topics added. In order to validate the content of the new course, local occupational physicians (OPs) were also surveyed as to their opinion of an appropriate curriculum in OEM for medical students. The new course was taught to the subsequent cohort of second year medical students. The students' ratings of the course pre- and post-revision were compared. The student satisfaction rates of the course significantly improved as a result of the changes. The content of the student-led curriculum was strikingly similar to the course proposed by the local OP with a few key exceptions. Student involvement in curriculum design in OEM is entirely feasible. It can result in a curriculum similar to that designed by expert opinion but has the advantage of strongly engaging student interest.


BioShuttle mobility in living cells studied with high-resolution FCS & CLSM methodologies.
With the increase in molecular diagnostics and patient-specific therapeutic approaches, the delivery and targeting of imaging molecules and pharmacologically active agents gain increasing importance. The ideal delivery system does not exist yet. The realization of two features is indispensable: first, a locally high concentration of target-specific diagnostic and therapeutic molecules; second, the broad development of effective and safe carrier systems. Here we characterize the transport properties of the peptide-based BioShuttle transporter using FFM and CLSM methods. The modular design of BioShuttle-based formulations results in a multi-faceted field of applications, also as a theranostic tool.


Knowledge infrastructure and safe operating spaces in social-ecological systems.
Maintaining safe operating spaces for exploited natural systems in the face of uncertainty is a key sustainability challenge. This challenge can be viewed as a problem in which human society must navigate in a limited space of acceptable futures in which humans enjoy sufficient well-being and avoid crossing planetary boundaries. A critical obstacle is the nature of society as a controller with endogenous dynamics affected by knowledge, values, and decision-making fallacies. We outline an approach for analyzing the role of knowledge infrastructure in maintaining safe operating spaces. Using a classic natural resource problem as an illustration, we find that a small safe operating space exists that is insensitive to the type of policy implementation, while in general, a larger safe operating space exists which is dependent on the implementation of the "right" policy. Our analysis suggests the importance of considering societal response dynamics to varying policy instruments in defining the shape of safe operating spaces.


Socioeconomic status and smoking: analysing inequalities with multiple indicators.
Socioeconomic differences in smoking have been well established. While previous studies have mostly relied on one socioeconomic indicator at a time, this study examined socioeconomic differences in smoking by using several indicators that reflect different dimensions of socioeconomic position. Data derive from Helsinki Health Study baseline surveys conducted among the employees of the City of Helsinki in 2000 and 2001. The data include 6243 respondents aged 40-60 years (response rate 68%). Six socioeconomic indicators were used: education, occupational status, household income per consumption unit, housing tenure, economic difficulties and economic satisfaction. Their associations with current smoking were examined by fitting sequential logistic regression models. All socioeconomic indicators were strongly associated with smoking among both men and women. When the indicators were examined simultaneously their associations with smoking attenuated, especially when education and occupational status were considered together, and when income and housing tenure were introduced into the models already containing education and occupational status. After mutual adjustment for all socioeconomic indicators, housing tenure and economic satisfaction remained associated with smoking in men. In women, all indicators except income and economic difficulties were inversely associated with smoking after adjustments. Smoking was associated with structural, material as well as perceived dimensions of socioeconomic disadvantage. Attempts to reduce smoking among the socioeconomically disadvantaged need to target several dimensions of socioeconomic position.


cryptochrome genes form an oscillatory loop independent of the per/tim loop in the circadian clockwork of the cricket Gryllus bimaculatus.
Animals exhibit circadian rhythms with a period of approximately 24 h in various physiological functions, including locomotor activity. This rhythm is controlled by an endogenous oscillatory mechanism, or circadian clock, which consists of cyclically expressed clock genes and their product proteins. cryptochrome (cry) genes are thought to be involved in the clock mechanism, and their functions have been examined extensively in holometabolous insects, but in hemimetabolous insects their role is less well understood. In the present study, the role of cry genes was investigated using RNAi technology in a hemimetabolous insect, the cricket Gryllus bimaculatus. Using a molecular cloning approach, we obtained cDNAs for two cry genes: Drosophila-type cry1 (Gb'cry1) and mammalian-type cry2 (Gb'cry2). Gb'cry2 has six splicing variants, most of which showed rhythmic mRNA expression. Gb'cry1RNAi treatment had only a limited effect at the behavioral and molecular levels, while Gb'cry2RNAi had a significant effect on behavioral rhythms and molecular oscillatory machinery, alone or in combination with Gb'cry1RNAi. In Gb'cry1/Gb'cry2 double-RNAi crickets, most clock genes showed arrhythmic expression, except for timeless, which retained clear rhythmic expression. Molecular analysis revealed that some combination of Gb'cry1 and Gb'cry2 variants suppressed CLK/CYC transcriptional activity in cultured cells. Based on these results, we propose a new model of the cricket's circadian clock, including a molecular oscillatory loop for Gb'cry2, which can operate independent of the Gb'per/Gb'tim loop.


Managing military penetrating brain injuries in the war zone: lessons learned.
OBJECTIVEManaging penetrating military brain injuries in a war zone setting is different than managing common civilian penetrating brain injuries. Triage, i.e., deciding on which patients to treat or not treat, and which to be flown back home, is essential to avoid wasting valuable limited resources. In this study the authors aim to identify reliable predictors of mortality and poor outcome to help develop a protocol for treating their patients in the battlefield. They also demonstrate all the lessons learned from their collective experience regarding some of the controversial management issues.METHODSThis study was a retrospective review of 102 patients with penetrating military missile head injuries treated by the authors in various facilities in northern Sinai between 2011 and 2018. Patient demographics, clinical characteristics, imaging characteristics, postoperative complications, and Glasgow Outcome Scale (GOS) scores were recorded for each patient. Several variables associated with mortality and poor outcome that were derived from the literature were analyzed, in addition to variables obtained by direct observation by the authors over time.RESULTSThere were 50 patients (49%) with GOS score of 1 (death), 12 patients (11.8%) with GOS score of 2 (survivors in persistent vegetative state), and 40 survivors (39.2%) with varying degrees of disability on the last follow-up evaluation. The authors identified an anatomical danger zone that was found to predict mortality when traversed. Bilateral dilated fixed pupils and low Glasgow Coma Scale score on admission were also found to be independent predictors of mortality and poor outcome. Based on these findings, a protocol was developed for managing these patients in the war zone.CONCLUSIONSManaging military penetrating head injuries in the war zone is different than civilian gunshot head injuries encountered by most neurosurgeons in urban cities. The authors developed a simple protocol for managing military penetrating injuries in the war zone. They also describe important lessons learned from this experience.


Tetraploid cells from cytokinesis failure induce aneuploidy and spontaneous transformation of mouse ovarian surface epithelial cells.
Most ovarian cancers originate from the ovarian surface epithelium and are characterized by aneuploid karyotypes. Aneuploidy, a consequence of chromosome instability, is an early event during the development of ovarian cancers. However, how aneuploid cells are evolved from normal diploid cells in ovarian cancers remains unknown. In the present study, cytogenetic analyses of a mouse syngeneic ovarian cancer model revealed that diploid mouse ovarian surface epithelial cells (MOSECs) experienced an intermediate tetraploid cell stage, before evolving to aneuploid (mainly near-tetraploid) cells. Using long-term live-cell imaging followed by fluorescence in situ hybridization (FISH), we demonstrated that tetraploid cells originally arose from cytokinesis failure of bipolar mitosis in diploid cells, and gave rise to aneuploid cells through chromosome mis-segregation during both bipolar and multipolar mitoses. Injection of the late passage aneuploid MOSECs resulted in tumor formation in C57BL/6 mice. Therefore, we reveal a pathway for the evolution of diploid to aneuploid MOSECs and elucidate a mechanism for the development of near-tetraploid ovarian cancer cells.


Dendritic Structures Functionalized with Boron Clusters, in Particular Carboranes, and Their Biological Properties.
The presence of a large number of boron atoms in boron clusters make them attractive tools for the treatment of cancer using boron neutron capture therapy (BNCT). Since the quantity of boron atoms present in the target cell directly affects the effectiveness of BNCT, the idea of gathering a high number of boron atoms in a single entity has emerged many years ago. In this perspective, using hyper-branched macromolecules such as dendrimers appears as an interesting solution. In this review, we will first present the synthesis of diverse dendritic entities (dendrimers, dendrons, and Janus dendrimers) that incorporate boron clusters, in particular carboranes, anywhere in their structure. Four parts of this review present the synthesis of dendrimers having boron clusters on the surface, or inside their structure, of dendrons and of Janus dendrimers, bearing boron clusters. Practically all these boronated dendritic structures were synthesized with the objective to study their biological properties, but in fact only a few of them have been tested against cancerous cells, and even a smaller number was tested in BNCT experiments. The biological experiments are discussed in the fifth part of this review. A good efficiency is generally observed with the boronated dendrimers, even in animal models, with an increase in their mean survival time (MST).


[Possibilities and results of organ-saving surgery in injuries of the spleen].
An analysis of 218 patients with spleen traumas treated surgically has been made. The organ-saving operations were made in 70 patients with a laser scalpel. The absolute and relative indications for such operations were determined. Complications and lethal outcomes in the early postoperative period in patients with the preserved spleen and after splenectomy were 21.1% and 12.9%, 30% and 13% correspondingly.


Evidence base and practice variation in acute care processes for knee and hip arthroplasty surgeries.
Lack of evidence contributes to unnecessary variation in treatment costs and outcomes. This study aimed to identify from interventions historically used for total knee or hip arthroplasty (TKA, THA): i) if routine use is supported by high-level evidence; ii) whether surgeon use aligns with the evidence. Part 1: Systematic search of electronic library databases for systematic reviews and practice guidelines concerning seven acute-care interventions. Intervention-specific recommendations concerning routine use were extracted by assessors. Part 2: Prospective medical record audit of the acute-care received by 1900 patients involving 120 orthopaedic surgeons. Surgeon use per intervention was summarized using caterpillar plots. Surgeon-specific routine and non-routine use was defined as use in ≥ 90% and ≤ 10% of patients, respectively. Primary analysis included only surgeons contributing ≥ 10 patients. Continuous passive motion (TKA): Routine use not recommended; 85.7% of surgeons did not use it routinely. Tranexamic Acid: Routine use recommended; 26.9% of surgeons used it routinely. Cryotherapy: Routine use not recommended; 45.7% of surgeons used it routinely for TKA; 31.8% used it routinely for THA. Intra-articular drainage: Routine use not recommended for TKA, but possible benefits for THA; 5.7% of surgeons used it routinely for TKA, 0.0% used it routinely for THA. Antibiotic loaded bone cement: Routine use for TKA not supported, recommendations for use for THA are inconsistent; 90.0% of surgeons used it routinely for TKA, 100.0% used it routinely for THA. Patella resurfacing (TKA): No recommendation could be made; 57.1% of surgeons routinely resurfaced the patella. Indwelling urinary catheterisation: Routine use recommended; 59.6% of surgeons used it routinely. Recommendations for routine use or not exist for some of the acute-care interventions examined. Surgeon practices vary widely even in the presence of high-level recommendations. It is unclear whether further evidence alone would lessen unwarranted practice variation.


The cost-effectiveness of controlling cervical cancer using a new 9-valent human papillomavirus vaccine among school-aged girls in Australia.
Cervical cancer imposes a substantial health burden worldwide including in Australia and is caused by persistent infection with one of 13 sexually transmitted high-risk human papillomavirus (HPV) types. The objective of this study was to assess the cost-effectiveness of adding a nonavalent new Gardasil-9® (9vHPV) vaccine to the national immunisation schedule in Australia across three different delivery strategies. The Papillomavirus Rapid Interface for Modelling and Economics (PRIME) model was used to examine the cost-effectiveness of 9vHPV vaccine introduction to prevent HPV infection. Academic literature and anecdotal evidence were included on the demographic variables, cervical cancer incidence and mortality, treatment costs, and vaccine delivery costs. The incremental cost-effectiveness ratios (ICERs) were measured per disability-adjusted life years (DALYs) averted, using the heuristic cost-effectiveness threshold defined by the World Health Organisation (WHO). Analyses and data from international agencies were used in scenario analysis from the health system and societal perspectives. The 9vHPV vaccination was estimated to prevent 113 new cases of cervical cancer (discounted) during a 20-year period. From the health system and societal perspectives, the 9vHPV vaccination was very cost-effective in comparison with the status quo, with an ICER of A$47,008 and A$44,678 per DALY averted, respectively, using the heuristic cost-effectiveness threshold level. Considering delivery strategies, the ICERs per DALY averted were A$47,605, A$46,682, and A$46,738 for school, health facilities, and outreach-based vaccination programs from the health system perspective, wherein, from the societal perspective, the ICERs per DALY averted were A$46,378, A$43,729, A$43,930, respectively. All estimates of ICERs fell below the threshold level (A$73,267). This cost-effectiveness evaluation suggests that the routine two-dose 9vHPV vaccination strategy of preadolescent girls against HPV is very cost-effective in Australia from both the health system and societal perspectives. If equally priced, the 9vHPV option is the most economically viable vaccine. Overall, this analysis seeks to contribute to an evidence-based recommendation about the new 9vHPV vaccination in the national immunisation program in Australia.


A novel role of the aryl hydrocarbon receptor (AhR) in centrosome amplification - implications for chemoprevention.
Centrosome aberrations can cause genomic instability and correlate with malignant progression in common human malignancies such as breast and prostate cancer. Deregulation of cyclin/cyclin-dependent kinase 2 (CDK2) activity has previously been shown to be critically involved in centrosome overduplication. We therefore test here whether small molecule CDK inhibitors derived from the bis-indole indirubin can be used to suppress centrosome aberrations as a novel approach to chemoprevention of malignant progression. As expected, we found that the CDK inhibitor indirubin-3'-oxime (IO) suppresses centrosome amplification in breast cancer cells. However, we made the unexpected discovery that indirubin-derived compounds that have been chemically modified to be inactive as kinase inhibitors such as 1-methyl-indirubin-3'-oxime (MeIO) still significantly reduced centrosome amplification. All indirubins used in the present study are potent agonists of the aryl hydrocarbon receptor (AhR), which is known for its important role in the cellular metabolism of xenobiotics. To corroborate our results, we first show that the coincidence of nuclear AhR overexpression, reflecting a constitutive activation, and numerical centrosome aberrations correlates significantly with malignancy in mammary tissue specimens. Remarkably, a considerable proportion (72.7%) of benign mammary tissue samples scored also positive for nuclear AhR overexpression. We furthermore provide evidence that continued expression of endogenous AhR is critical to promote centriole overduplication induced by cyclin E and that AhR and cyclin E may function in the same pathway. Overexpression of the AhR in the absence of exogenous ligands was found to rapidly disrupt centriole duplication control. Nonetheless, the AhR agonists IO and MeIO were still found to significantly reduce centriole overduplication stimulated by ectopic AhR expression. Our results indicate that continued expression of endogenous AhR promotes centrosome amplification in breast cancer cells in a pathway that involves cyclin E. AhR agonists such as indirubins inhibit centrosome amplification even when stimulated by ectopic expression of the AhR suggesting that these compounds are potentially useful for the chemoprevention of centrosome-mediated cell division errors and malignant progression in neoplasms in which the AhR is overexpressed. Future studies are warranted to determine whether individuals in which nuclear AhR overexpression is detected in benign mammary tissue are at a higher risk for developing pre-cancerous or cancerous breast lesions.


Parental contexts of adolescent self-esteem: A developmental perspective.
Relationships between parental behaviors and adolescent self-esteem were analyzed in a group of 95 early adolescents from multiple settings. The study was designed to investigate hypotheses regarding associations between observed parental interactions (e.g., accepting and devaluing) and adolescent self-esteem. Parents' verbal interactions with their adolescents were assessed through application of the constraining and enabling coding system to transcribed family discussions, generated through a revealed differences procedure. Adolescent self-esteem was measured with the Coopersmith Self-Esteem Inventory. Parent interaction-self-esteem associations were examined in the pooled sample, as well as in specific sub-groups based on gender, health, and ego development (measured by the Washington University Sentence Completion Test). Boys had more numerous associations between their self-esteem and parental interactions than girls, and psychiatrically ill boys had particularly high associations. Parental interactions were found to be most strongly related to adolescent self-esteem for adolescents at the lowest levels of ego development. Our findings are consistent with the view that increasing individuation in self-esteem regulation occurs during adolescent development, such that adolescents at higher levels of ego development evaluate themselves more independently of parental feedback than do their less mature peers.


Outer retinal degeneration: an electronic retinal prosthesis as a treatment strategy.
To review progress toward an electronic retinal prosthesis for outer retinal degeneration. Literature review. Retinal degenerations such as retinitis pigmentosa result in a loss of photoreceptors. There is a secondary loss of inner retinal cells, but significant numbers of bipolar and ganglion cells remain for many years. Electrical stimulation can produce phosphenes in the eyes of individuals who are blind as a result of retinitis pigmentosa. Several research groups are trying to exploit this phenomenon to produce artificial vision with electronic retinal prostheses. Two groups, with private company sponsorship, have recently implanted first-generation devices in subjects with advanced retinitis pigmentosa. They have reported limited preliminary results. This article seeks to put these results in a broader context and review potential obstacles to successful prosthesis development. These include inner retinal cell viability, high thresholds, signal encoding, power requirements, biocompatibility, and device encapsulation. There has been substantial progress toward an electronic retinal prosthesis, but fully functional, long-lasting devices are not on the immediate horizon.


Effects of the PCSK9 antibody alirocumab on coronary atherosclerosis in patients with acute myocardial infarction: a serial, multivessel, intravascular ultrasound, near-infrared spectroscopy and optical coherence tomography imaging study-Rationale and design of the PACMAN-AMI trial.
The risk for cardiovascular adverse events after acute myocardial infarction (AMI) remains high despite potent medical treatment including low-density lipoprotein cholesterol (LDL-C) lowering with statins. Proprotein convertase subtilisin/kexin type 9 (PCSK9) antibodies substantially reduce LDL-C when added to statin. Alirocumab, a monoclonal antibody to PCSK9, reduces major adverse cardiovascular events after AMI. The effects of alirocumab on coronary atherosclerosis including plaque burden, plaque composition and fibrous cap thickness in patients presenting with AMI remains unknown. To determine the effect of LDL-C lowering with alirocumab on top of high-intensity statin therapy on intravascular ultrasound (IVUS)-derived percent atheroma volume (PAV), near-infrared spectroscopy (NIRS)-derived maximum lipid core burden index within 4 mm (maxLCBI4 mm) and optical coherence tomography (OCT)-derived fibrous cap thickness (FCT) in patients with AMI. In this multicenter, double-blind, placebo-controlled trial, 300 patients with AMI (ST-elevation or non-ST-elevation myocardial infarction) were randomly assigned to receive either biweekly subcutaneous alirocumab (150 mg) or placebo beginning <24 hours after the acute event as add-on therapy to rosuvastatin 20 mg. Patients undergo serial IVUS, NIRS and OCT in the two non-infarct related arteries at baseline (at the time of treatment of the culprit lesion) and at 52 weeks. The primary endpoint, change in IVUS-derived PAV, and the powered secondary endpoints, change in NIRS-derived maxLCBI4 mm, and OCT-derived minimal FCT, will be assessed 52 weeks post randomization. The PACMAN-AMI trial will determine the effect of alirocumab on top of high-intensity statin therapy on high-risk coronary plaque characteristics as assessed by serial, multimodality intracoronary imaging in patients presenting with AMI. NCT03067844.


Comparing the Effects of Sleep and Rest on Memory Consolidation.
There is ample evidence that overnight sleep and daytime naps benefit memory retention, compared to comparable amounts of active wakefulness. Yet recent evidence also suggests that a period of post-training rest (eg, quiet wakefulness with eyes closed) provides a similar memory benefit compared to wake. However, the relative benefits of sleep vs quiet waking rest on memory remain poorly understood. Here, we assessed the extent to which sleep provides a unique memory benefit, above and beyond that conferred by quiet waking rest. In a sample of healthy undergraduate students (N=83), we tested the effect of 30 mins of post-learning sleep, rest, or active wake on concept learning (dot pattern classification) and declarative memory (word pair associates) across a 4-hr daytime training-retest interval. Contrary to our hypotheses, we found no differences in performance between the three conditions for either task. The findings are interpreted with reference to methodological considerations including the length of the experimental interval, the nature of the tasks used, and challenges inherent in creating experimental conditions that can be executed by participants.


[Pertussis hospitalisations: Northern Region of Portugal, 2000-2006].
Pertussis is an acute bacterial disease involving the respiratory tract. The causal agent is Bordetella pertussis. Among all age groups, infants (aged < 12 months) have had the highest incidence of pertussis in the vaccine era. The majority of hospitalisations, complications and deaths occurred in that age group. The aim of this study was to describe and characterize the epidemiological pattern of hospitalized pertussis cases during the period of 2000-2006 in the hospitals of Northern Portugal. We conducted a retrospective observational study based on hospitalisations in the Northern region of Portugal between 2000 and 2006. Descriptive summary statistics for all variables were calculated at region, district and municipality levels. The average rates of hospitalisation were calculated per 100,000 population for the region, district and municipality, according to sex and age group. The severity of disease was evaluated on the basis of the average length of hospitalisation, the proportion of patients treated in intensive care units, and the case fatality rate. Three hundred and twenty two cases were identified between 2000 and 2006 in the Northern region of Portugal. Viana do Castelo and Braga districts were those who had the highest hospitalisations rate. In the district of Bragança there were no cases of hospitalisation with the diagnosis of pertussis. Fifty six per cent of the cases were male. The proportion of infants hospitalized under one year was 91.0 %. Cases among infants younger than two months accounted for 43.5 % of all cases. The mean annual hospitalisation incidence rate for infants under one year was 114 per 100,000. The mean length of hospital stay was eight days. The proportion of patients treated in intensive care units was 11.0 %, and case fatality rate was 0.3 %. This hospital-based study indicates that pertussis continues to be an important cause of morbidity and mortality in infants. Pertussis incidence remains high among infants, most of whom are under one year of age. Immunization strategies in the adult population must be reviewed and updated in order to attain higher protection of the more vulnerable paediatric population.


Shifts in Lachnospira and Clostridium sp. in the 3-month stool microbiome are associated with preschool age asthma.
Asthma is a chronic disease of the airways affecting one in ten children in Westernized countries. Recently, our group showed that specific bacterial genera in early life are associated with atopy and wheezing in 1-year-old children. However, little is known about the link between the early life gut microbiome and the diagnosis of asthma in preschool age children. To determine the role of the gut microbiota in preschool age asthma, children up to 4 years of age enrolled in the Canadian Healthy Infant Longitudinal Development (CHILD) study were classified as asthmatic (n=39) or matched healthy controls (n=37). 16S rRNA sequencing and quantitative PCR (qPCR) were used to analyse the composition of the 3-month and 1-year gut microbiome of these children. At 3 months the abundance of the genus, Lachnospira (L), was decreased (P=0.008), whereas the abundance of the species, Clostridium neonatale (C), was increased (P=0.07) in asthmatics. Quartile analysis of stool composition at 3-months revealed a negative association between the ratio of these two bacteria (L/C) and asthma risk by 4 years of age [quartile 1: odds ratio (OR)=15, P=0.02, CI (confidence interval)= 1.8-124.7; quartile 2: OR=1.0, ns; quartile 3: OR=0.37, ns]. We conclude that opposing shifts in the relative abundances of Lachnospira and C. neonatale in the first 3 months of life are associated with preschool age asthma, and that the L/C ratio may serve as a potential early life biomarker to predict asthma development.


Innate Immune Sensing of Fusarium culmorum by Mouse Dendritic Cells.
Chronic inhalation of grain dust is associated with asthma and chronic bronchitis in grain worker populations. Exposure to fungal particles was postulated to be an important etiologic agent of these pathologies. Fusarium species frequently colonize grain and straw and produce a wide array of mycotoxins that impact human health, necessitating an evaluation of risk exposure by inhalation of Fusarium and its consequences on immune responses. Data showed that Fusarium culmorum is a frequent constituent of aerosols sampled during wheat harvesting in the Vaud region of Switzerland. The aim of this study was to examine cytokine/chemokine responses and innate immune sensing of F. culmorum in bone-marrow-derived dendritic cells and macrophages. Overall, dendritic cells and macrophages responded to F. culmorum spores but not to its secreted components (i.e., mycotoxins) by releasing large amounts of macrophage inflammatory protein (MIP)-1α, MIP-1β, MIP-2, monocyte chemoattractant protein (MCP)-1, RANTES, and interleukin (IL)-12p40, intermediate amounts of tumor necrosis factor (TNF), IL-6, IL-12p70, IL-33, granulocyte colony-stimulating factor (G-CSF), and interferon gamma-induced protein (IP-10), but no detectable amounts of IL-4 and IL-10, a pattern of mediators compatible with generation of Th1 or Th17 antifungal protective immune responses rather than with Th2-dependent allergic responses. The sensing of F. culmorum spores by dendritic cells required dectin-1, the main pattern recognition receptor involved in β-glucans detection, but likely not MyD88 and TRIF-dependent Toll-like receptors. Taken together, our results indicate that F. culmorum stimulates potently innate immune cells in a dectin-1-dependent manner, suggesting that inhalation of F. culmorum from grain dust may promote immune-related airway diseases in exposed worker populations.


Detection and residence time of bisphosphonates in bone of horses.
Bisphosphonates are potent anti-resorptive agents that have the potential to adversely affect bone healing in equine athletes, and normal bone adaption in young racehorses. A concern exists that bisphosphonate inhibition of normal bone metabolism could lead to increased bone fractures during high-intensity exercise. We found only a single report describing concentrations of tiludronate in the bone of horses, and no studies describing clodronate. Knowledge of the residence time in bone could allow for a better understanding of the long-term effects of these compounds. Our objectives were to develop a method for detection of bisphosphonates in bone and add to the limited information available regarding the disposition of these drugs in the bone of horses. Two horses received clodronate and 2 tiludronate disodium. Postmortem collection of bones and teeth occurred either 4 or 30 d post drug administration. Additionally, postmortem blood, synovial fluid, aqueous humor, and bone samples from racehorses with various histories of bisphosphonate administration were collected, and concentrations determined using the developed LC-MS/MS method. Bisphosphonates were detected in bones and teeth tested at 4 and 30 d. In a postmortem sample, clodronate was detected in bone from a horse with reported administration 18 mo prior; clodronate was not detected in other sample types collected from this horse. Bisphosphonates reside in bone for extended periods of time, which could lead to potential long-term effects, increasing the potential for bone fractures in young and/or athletic horses.


[Proteomic analysis of differentially expressed proteins in human hepatoma SMMC-7721 cells induced by Fufang Banmao capsule serum].
To investigate the effects of Fufang Banmao capusle on the proteome of SMMC-7721 cells and discover potential molecular mechanism of anti-cancer at molecular level. SMMC-7721 cells were treated by Fufang Banmao capusle serum prepared with serum pharmacological method; proteomic protocol involving 2-DE, image analysis and mass spectrometry were used to detect the proteins in cells influenced by Fufang Banmao capusle. Approximately 450 protein spots in SMMC-7721 cells were resolved and detected in 2-D gel maps from pH3-10L IEF. 47 protein plots varied over 2-fold quantitively between treated sample and control sample were uncovered. 13 differentially expressed proteins spots were further identified by MALDI-TOF-MS analysis and four of them were successfully identified. Annexin A5, heatshock 70 x 10(3) protein 8 was significantly up-regulated in treated sample compared with control sample, while Eukaryotic translation initiation factor 5A and Peroxiredoxin-2 was significantly down-regulated in treated sample. 4 differently expressed proteins associated with the proliferation, apoptosis, immunity of tumor were detected and they might provide clues for the coming research. The protocol of proteomics combined with serum pharmacological method is an effective platform to research complicated formulas in that it is capable of laying out many proteins associated with Fufang Banmao capusle.


Reconsidering the roles of gratitude and indebtedness in social exchange.
Receiving favors is often a mixed blessing and commonly triggers two emotions: the positive emotion gratitude and negative emotion indebtedness. In three studies, we examined the hypothesis that gratitude and indebtedness have distinct functions in social exchange. Contrary to current views, we believe that the function of gratitude does not primarily reside in facilitating social exchange. Instead, we propose that indebtedness motivates people to repay favours received, and thus accounts for most of the prosocial effects commonly attributed to gratitude. On the other hand consistent with current views, we believe that gratitude signals the potential for developing a relationship and fosters proximity seeking. Supporting these assumptions, in Study 1 we found that gratitude and indebtedness were associated with aspects of the favour that reflect the concern for relationship and the level of inequity. Studies 2 and 3 provided causal support for these relations, and revealed the unique associations between gratitude and the motivation of proximity seeking, and between indebtedness and the motivation to reciprocate. We argue that this functional distinction has escaped research attention as gratitude and indebtedness are naturally correlated because they stem from the same eliciting event. To appreciate this functional distinction, both emotions should be studied simultaneously in the context of social exchange.


Qualitative Exploration of Engaging Patients as Advisors in a Program of Evidence Synthesis: Cobuilding the Science to Enhance Impact.
There is an increasing expectation for research to involve patient stakeholders. Yet little guidance exists regarding patient-engaged research in evidence synthesis. Embedded in a learning health care system, the Veteran Affairs Evidence Synthesis Program (ESP) provides an ideal environment for exploring patient-engaged research in a program of evidence synthesis. The objective of this study was to explore views, barriers, resources, and perceived values of engaging patient advisors in a national program of evidence synthesis research. We conducted 10 qualitative interviews with ESP researchers and 2 focus groups with patient stakeholder informants. We queried for challenges to patient involvement, resources needed to overcome barriers, and perceived values of patient engagement. We analyzed qualitative data using applied thematic and matrix techniques. Patient stakeholders and researchers expressed positive views on the potential role for patient engagement in the Veteran Affairs ESP. Possible contributions included topic prioritization, translating findings for lay audiences, and identifying clinically important outcomes relevant to patients. There were numerous barriers to patient involvement, which were more commonly noted by ESP researchers than by patient stakeholders. Although informants were able to articulate multiple values, we found a lack of clarity around measurable outcomes of patient involvement in systematic reviews. The research community increasingly seeks patient input. There are many perceived and actual barriers to seeking robust patient engagement in systematic reviews. This study outlines emerging practices that other evidence synthesis programs should consider, such as the careful selection of stakeholders; codeveloped expectations and goals; and adequate training and appropriate resources to ensure meaningful engagement.


Deletion of ghrelin impairs neither growth nor appetite.
Pharmacological studies show that ghrelin stimulates growth hormone release, appetite, and fat deposition, but ghrelin's physiological role in energy homeostasis has not been established. Ghrelin was also proposed to regulate leptin and insulin release and to be important for the normal function of stomach, heart, kidney, lung, testis, and placenta. To help determine a definable physiological role for ghrelin, we generated ghrelin-null mice. In contrast to predictions made from the pharmacology of ghrelin, ghrelin-null mice are not anorexic dwarfs; their size, growth rate, food intake, body composition, reproduction, gross behavior, and tissue pathology are indistinguishable from wild-type littermates. Fasting produces identical decreases in serum leptin and insulin in null and wild-type mice. Ghrelin-null mice display normal responses to starvation and diet-induced obesity. As in wild-type mice, the administration of exogenous ghrelin stimulates appetite in null mice. Our data show that ghrelin is not critically required for viability, fertility, growth, appetite, bone density, and fat deposition and not likely to be a direct regulator of leptin and insulin. Therefore, antagonists of ghrelin are unlikely to have broad utility as antiobesity agents.


Alternative futures for world cereal and meat consumption.
Fundamental changes in the global structure of food demand will lead to an extraordinary increase in the importance of developing countries in global food markets. Economic growth in developing countries is changing consumption patterns, with slower growth (and in many countries actual declines) in per capita food consumption of grains and rapidly growing per capita and total meat consumption, combined with induced growth in cereal feed consumption. The present paper examines the hypothesis, suggested by some researchers, that high-meat diets in developed countries limit improvement in food security in developing countries. These analysts argue that reduced meat consumption in developed countries would release cereals from livestock feed to food for poorer populations, thus improving food security in developing countries. Using the International Food Policy Research Institute (Washington, DC, USA) global food projections model, the international model for policy analysis of agricultural commodities and trade (see Rosegrant et al. 1995), we first analyse the implications for future global cereal and meat supply and demand resulting from changes in global income, population growth and other structural changes, then simulate alternative scenarios to examine the effect of large reductions in meat consumption in developed countries on food consumption and food security in developing countries. The paper shows that while the long-term prospects for food supply, demand and trade indicate a strengthening of world cereal and livestock markets, the improvement in food security in the developing world will be slow, and changes in the dietary patterns in developed countries are not an effective route to improvement in food security in developing countries.


Cloning, sequencing and characterisation of a Listeria monocytogenes gene encoding a fibronectin-binding protein.
Listeria monocytogenes is a gram-positive, non-sporulating food-borne pathogen of man and animals that is able to invade many eukaryotic cells. L. monocytogenes possesses several proteins that bind fibronectin. In this study, an L. monocytogenes DNA library in pUC19 was screened with fibronectin and a gene encoding a 24.6-kDa fibronectin-binding protein (Fbp) was isolated and sequenced. Transcripts of the fbp gene were found in wild-type, in deltaprfA, and PrfA-S183A strains, despite the presence of a 'PrfA-like' box around its ribosome-binding site. The fbp gene was found to be present in all tested isolates of the species L. monocytogenes and a homologous DNA fragment was amplified in L. welshimeri. No homologies between the fbp gene and its translation product with any other DNA or proteins deposited in databanks were found. Restriction endonuclease-PCR (RE-PCR) showed that the fbp gene displays a degree of allelic variation among isolates of L. monocytogenes, whereas the corresponding amplified fragment of L. welshimeri seems to be monomorphic among isolates of this species. RE-PCR with Hha I, Dde I or Taq I produced DNA banding profiles specific for each of these two species, allowing their identification.


In vitro measurement of glucose uptake after radiation and cetuximab treatment in head and neck cancer cell lines using 18F-FDG, gamma spectrometry and PET/CT.
The standard treatment for head and neck squamous cell carcinoma (HNSCC) is radiotherapy, often in combination with chemotherapy or surgery. However, a novel monoclonal antibody, cetuximab (Erbitux®), has also been approved for patient therapy. The aim of present study was to develop an in vitro method for the measurement of 18F-fluoro-2deoxy-D-glucose (FDG) to determine if cellular 18F-FDG uptake is associated with response to radiotherapy or cetuximab treatment. In the current study, HNSCC cell lines were treated with radiation or with cetuximab. Next, the uptake of 18F-FDG was measured using a gamma spectrometer (GS). Thereafter, uptake after radiation was measured first with GS and then compared with positron emission tomography (PET)/computed tomography (CT) imaging. Furthermore, the mRNA expression of glucose transporter 1 (GLUT1) was measured following cetuximab treatment via reverse transcription-quantitative PCR. A study protocol was developed to measure the cellular uptake of 18F-FDG via gamma-ray spectrometry and comparable results were obtained with those of clinical PET/CT. The results revealed a decrease in 18F-FDG after radiation and cetuximab treatment. The uptake of 18F-FDG following cetuximab treatment was significantly lower in the cetuximab-sensitive cell line UT-SCC-14 compared with the cetuximab-resistant cell lines UT-SCC-2 and UT-SCC-45. Furthermore, after treatment with cetuximab for 24 and 48 h, a significant increase in GLUT1 expression was detected in the sensitive cell line compared with the two resistant cell lines. In conclusion, a novel yet reliable method for the measurement of intracellular 18F-FDG via GS has been developed, and our results indicate that 18F-FDG uptake is associated with radiation and cetuximab response in HNSCC.


Increased mortality from infectious pneumonia after occupational exposure to inorganic dust, metal fumes and chemicals.
There are epidemiological studies indicating that exposure to metal fumes is a risk factor for infectious pneumonia. Whether occupational exposure to other agents, such as inorganic dust or chemicals, also increases the risk for infectious pneumonia is not clear. The aim of the present study was to elucidate whether occupational exposure to respiratory pollutants and irritants increases the risk for infectious pneumonia. Prospective cohort study. Setting Swedish male construction workers. Participants 320,143 male construction workers exposed to inorganic dust (asbestos, man-made mineral fibres, dust from cement, concrete and quartz), wood dust, metal fumes and chemicals (organic solvents, diisocyanates and epoxi resins) or unexposed. Main outcome measures The cohort was followed from 1971 to 2003 and the main outcome measures were mortality to infectious pneumonia, lobar pneumonia or pneumococcal pneumonia. RRs were obtained by the person-years method and from Poisson regression models, adjusting for baseline values of age and smoking habits. Among men aged 20-64 years there was increased mortality from infectious pneumonias among construction workers exposed to metal fumes (RR 2.31, 95% CI 1.35 to 3.95), inorganic dust (RR 1.87, 95% CI 1.22 to 2.87) and chemicals (RR 1.91, 95% CI 1.37 to 3.22). The mortality was also increased from both lobar pneumonia and pneumococcal pneumonia. Among men aged 65-84 years the occupational exposure to inorganic dust and chemicals was associated with slightly increased mortality from infectious pneumonia. Among groups with mutually exclusive exposures there was increased mortality from infectious pneumonias among construction workers exposed to inorganic dust, but not among those exposed to wood dust or chemicals. There were no cases among workers exposed only to metal fumes. Our findings indicate that exposure to inorganic dust increases the mortality from infectious pneumonias, especially lobar pneumonia and pneumococcal pneumonia. The mechanism is unclear, but the effect may be mediated through induced airways inflammation.


The role of the macrophage in genetic control of the immune response.
The ability of an animal to mount an immune response is controlled by a number of autosomal dominant immune response (Ir) genes that are linked to the major histocompatibility complex of the species. In the guinea pig, alloantiserums raised by cross-immunization of inbred strain 2 and strain 13 animals specifically inhibited the in vitro proliferative responses of (2 X 13) F1 lymphocytes to those antigens the response to which is controlled by Ir genes linked to the alloantigens against which the serums are directed. A genetic analysis indicated that the inhibitory activity of the alloantiserums was directed against the alloantigens rather than the products of specific Ir genes. The interaction of antigen-pulsed macrophages with immune T lymphocytes is also mediated by the 2/13 alloantigens and alloantiserums are capable of inhibiting macrophage-T lymphocyte interaction. Studies involving combinations of macrophages and lymphocytes that differed at alloantigens or Ir gene products or both raised the possibility of the expression of the Ir gene product in the macrophage.


Cellulolytic, fermentative, and methanogenic guilds in benthic periphyton mats from the Florida Everglades.
Phosphorus enrichment caused by runoff from agricultural areas has resulted in ecosystem-level changes in the northern Florida Everglades, including a loss of periphyton mats from nutrient-impacted areas. The potential for methanogenesis resulting from the anaerobic decomposition of cellulose and fermentation products, and the microorganisms responsible for these processes, were studied in mats from a region not impacted by nutrient enrichment. Methane was produced from periphyton incubated with cellulose, propionate, butyrate, and formate, with an accumulation of fatty acids in incubations. The accumulation of fatty acids may have been caused by the inhibition of syntrophic oxidation, a potentially significant route for methane production in soils. Sequence analysis of 16S rRNA genes characteristic of Clostridium, the primary genus responsible for anaerobic decomposition and fermentation in soils of the area, indicated that Clostridium Cluster I assemblages present in the mat differed from those in the soils of the area. Significantly, sequences characteristic of the Clostridium group that dominates the soils of the area, group XIV, were not detected in the mat. These results indicate that benthic periphyton is probably a significant source of methane in the Everglades, and the responsible microorganisms differ significantly from those in the soils of the area.


Correction of glycogenosis type 2 by muscle-specific lentiviral vector.
Glycogen storage disease type II (GSDII) or Pompe disease is an inherited disease of glycogen metabolism caused by a lack of functional lysosomal acid alpha-glucosidase (GAA). Affected individuals store glycogen in lysosomes resulting in fatal hypertrophic cardiomyopathy and respiratory failure in the most severe form. We investigated for the first time the use of lentiviral vectors to correct the GSDII phenotype in human and murine GAA-deficient cells. Fibroblasts from infantile and adult GSDII patients were efficiently transduced by a GAA-expressing lentiviral vector placed under the control of the strong MND promoter, leading to a complete restoration of enzymatic activity. We also developed a muscle-specific lentiviral vector based on the synthetic C5-12 promoter and tested it on deficient myogenic satellite cells derived from a GSDII mouse model. GAA was expressed as a correctly processed protein allowing a complete enzymatic and metabolic correction in myoblasts and differentiated myotubes, as well as a significant mannose-6-phosphate (M6P)-dependent secretion reuptake by naive cells. Transduced cells showed lysosomal glycogen clearance, as demonstrated by electron microscopy. These results form the basis for a therapeutic approach of GSDII using lentiviral vector-mediated gene transfer into muscle stem cells.


[Are the results of intragastric pH-metry and measurements of the pH of extracted gastric juice comparable?].
Fundal pH and pH of withdrawn gastric juice were measured under basal condition (with no stimulants used) in normal subjects and patients with chronic moderately severe and marked fundal gastritis and duodenal ulcer (altogether 96 examinees). Aspiration pH-metry with a tube designed by Ye. Yu. Linar was employed. Comparison of the results revealed statistically significant differences (the 'agreement criterion') in the pH values obtained by the compared methods. Low pH values were more frequent in intragastric fundal pH-metry, possibly at the expense of the presence of not only acid but of the antral-alkaline component in the withdrawn gastric juice. These differences should be taken into consideration in the clinical laboratory diagnosis.


Simultaneous Visualization of Multiple Gene Expression in Single Cells Using an Engineered Multicolor Reporter Toolbox and Approach of Spectral Crosstalk Correction.
Synthetic biology aims to make biology easier to engineer and focuses on the design and construction of core components that can be modeled, understood, and tuned to meet specific performance criteria, and the assembly of these smaller parts and devices into larger integrated systems to solve specific problems. Here, we designed and engineered a multicolor fluorescent reporter toolbox to simultaneously monitor the activities of multiple genes in single cells. The toolbox contained standardized and well-characterized genetic building blocks for the convenient and reproducible assembly of multiple promoter-reporter fusions (ranging from 1 to 4) into a single plasmid. Given the common problem of spectral crosstalk among multiple fluorescent proteins, we deciphered multiple spectral signatures within cells through a deduced linear unmixing algorithm. Our approach enabled the quantification of gene expression with direct FP concentrations, instead of mix-contributed fluorescence intensities, thus enabling true signal separation with high confidence. This approach performed well in the imaging of mixing cells with single FP labels. Additionally, combining with the multicolor toolbox, we succeeded in simultaneously monitoring the genetic dynamics of four selected quorum-sensing genes in response to the induction of two exogenously added autoinducers and were able to examine gene regulatory connections within the QS signaling network in Pseudomonas aeruginosa. Overall, this synthetic framework (i.e., the genetic toolbox and the well-evaluated approach of spectral correction) will be useful for applied synthetic biology projects, multicolor imaging, and analyzing interactions of multiple genes of natural genetic networks or assembling synthetic ones.


Can physical activity improve the mental health of older adults?
The world population is aging rapidly. Whilst this dramatic demographic change is a desirable and welcome phenomenon, particularly in view of people's increasing longevity, it's social, financial and health consequences can not be ignored. In addition to an increase of many age related physical illnesses, this demographic change will also lead to an increase of a number of mental health problems in older adults and in particular of dementia and depression. Therefore, any health promotion approach that could facilitate introduction of effective primary, secondary and even tertiary prevention strategies in old age psychiatry would be of significant importance. This paper explores physical activity as one of possible health promotion strategies and evaluates the existing evidence that supports its positive effect on cognitive impairment and depression in later life.


Out-of-pocket payments for public healthcare services by selected exempted groups in Serbia during the period of post-war healthcare reforms.
This paper focuses on the exemption mechanism that accompanies patient co-payments for outpatient and inpatient hospital care in Serbia. The objective was to investigate the level and dynamics of out-of-pocket payments for this type of services by exempted groups (older than 65 years, younger than 15 years, unemployed, disabled and individuals with low family income) compared with that by other groups. For this purpose, we use empirical household data collected in the Serbian Living Standards Measurement Study carried out in 2002, 2003 and 2007. These years correspond to the start of the recent reforms in the Serbian healthcare sector and 1 and 5 years after the start of the reform. Our results show that people who belong to exempted groups were paying for healthcare in 2002, 2003 and 2007. They report different types of out-of-pocket payments for outpatient and inpatient hospital care. Thus, despite the ambition of the Ministry of Health in Serbia to promote equity in healthcare as a leading aim of the reforms, the implementation of the exemption mechanism fails to protect the targeted groups. Future exemption mechanism should be pro-poor oriented but should also exempt those whose health status requires a frequent healthcare use.


Enhanced low-rank representation via sparse manifold adaption for semi-supervised learning.
Constructing an informative and discriminative graph plays an important role in various pattern recognition tasks such as clustering and classification. Among the existing graph-based learning models, low-rank representation (LRR) is a very competitive one, which has been extensively employed in spectral clustering and semi-supervised learning (SSL). In SSL, the graph is composed of both labeled and unlabeled samples, where the edge weights are calculated based on the LRR coefficients. However, most of existing LRR related approaches fail to consider the geometrical structure of data, which has been shown beneficial for discriminative tasks. In this paper, we propose an enhanced LRR via sparse manifold adaption, termed manifold low-rank representation (MLRR), to learn low-rank data representation. MLRR can explicitly take the data local manifold structure into consideration, which can be identified by the geometric sparsity idea; specifically, the local tangent space of each data point was sought by solving a sparse representation objective. Therefore, the graph to depict the relationship of data points can be built once the manifold information is obtained. We incorporate a regularizer into LRR to make the learned coefficients preserve the geometric constraints revealed in the data space. As a result, MLRR combines both the global information emphasized by low-rank property and the local information emphasized by the identified manifold structure. Extensive experimental results on semi-supervised classification tasks demonstrate that MLRR is an excellent method in comparison with several state-of-the-art graph construction approaches.


New insights into the vancomycin-induced nephrotoxicity using in vitro metabolomics combined with physiologically based pharmacokinetic modeling.
Vancomycin is a first-line treatment for invasive infections caused by multidrug-resistant gram-positive bacteria. However, vancomycin-induced nephrotoxicity is an increasing burden, particularly in patients with complex life-threatening conditions. Vancomycin-induced nephrotoxicity associated with clinically relevant exposure on the target site has not been well defined. This study aimed to acquire the concentration of vancomycin in the renal tubules and kidneys in humans using physiologically based pharmacokinetic (PBPK) modeling and simulation. Based upon the exposure of vancomycin in the renal tubule, the toxicity of vancomycin in human renal proximal tubular epithelial cells was examined with the XTT assay and in vitro metabolomics analysis. A rat PBPK model predicting plasma and kidney concentration-time profiles of vancomycin matched the observed behavior after a single administration of 10 mg/kg. The concentration of vancomycin in renal tubules was about 40-50 times higher than that in plasma. The human PBPK model transferred from the rat model predicted renal tubule concentrations of vancomycin as 316.1-2136.6 μg/mL at 500 mg every 6 hours, and 199.0-3932.5 μg/mL at 1000 mg every 12 hours. Vancomycin showed significant nephrotoxicity at 4 mg/mL in XTT assessment. In total, 11 lysophosphatidylcholines and one lysophosphatidylethanolamine were identified by metabolomics analysis. The concentration-dependent increase was evident in the release of lysophospholipids after vancomycin treatment (0.125-4 mg/mL) for 24 hours. Our study revealed the relationship between the exposure of vancomycin in the kidney and toxicity of vancomycin at clinically relevant concentrations achieved from a mechanical PBPK model. A series of lysophospholipids as potential metabolic markers of renal toxicity were identified.


Age of Canada basin deep waters: a way to estimate primary production for the arctic ocean.
An empirical model of carbon flux and (14)C-derived ages of the water in the Canada Basin of the Arctic Ocean as a function of depth was used to estimate the long-term rate of primary production within this region. An estimate can be made because the deep waters of the Canadian Basin are isolated from the world oceans by the Lomonosov Ridge (sill depth about 1500 meters). Below the sill, the age of the water correlates with increased nutrients and oxygen utilization and thus provides a way to model the average flux of organic material into the deep basin over a long time period. The (14)C ages of the deep water in the Canada Basin were about 1000 years, the carbon flux across the 1500-meter isobath was 0.3 gram of carbon per square meter per year, and the total production was 9 to 14 grams of carbon per square meter per year. Such estimates provide a baseline for understanding the role of the Arctic Ocean in global carbon cycling.


Retroperitoneoscopic hand-assisted live-donor nephrectomy according to the basic principle of transplantation in donor kidney selection.
We assessed the feasibility of retroperitoneoscopic hand-assisted live-donor nephrectomy according to the basic principle of transplantation in kidney selection, namely, leaving the better-functioning kidney in the donor. Thirty consecutive live-donor nephrectomies, including 10 right-sided and 20 left-sided procedures, were evaluated. The surgery was started endoscopically using three ports, followed by hand assistance for dissecting the renal pedicles through the extended inner-port incision. A vascular Endostapler and polymer clips were used to transect the renal vessels. Two right-sided cases required open conversion because of multiple renal vessels and uncontrollable bleeding. The median operative time, warm ischemia time (WIT), blood loss, and renal vein length were 244 minutes (upper and lower quartile 215 and 274 minutes), 186 seconds (134, 239 seconds), 175 mL (45, 305 mL), and 22 mm (19, 26 mm), respectively. The operative time and WIT were longer, and the renal vein was shorter, in the right-sided than in the left-sided procedures (P < 0.05), but no difference was found in the other perioperative data for the two sides. No delayed graft function was observed, and the kidney function 1 month postoperatively was acceptable in all donors and all recipients. Our technical devices, such as the site and timing of hand assistance and control of the renal vessels, seem feasible. Although we could not draw a conclusion about the safety of the right-sided procedure, this alternative procedure should be applicable for laparoscopic donor nephrectomy uninfluenced by the side of the donor kidney provided the surgical team has sufficient expertise.


Separation of lipid classes by solid phase extraction.
A rapid and reliable method for the separation of lipid classes is described using aminopropyl disposable columns. This method is a modification to an existing procedure that allows the separation of both neutral and acidic phospholipid fractions and a high recovery of the latter. Acidic phospholipids were eluted with a mixture of hexane-2-propanol-ethanol-0.1 M ammonium acetate-formic acid 420:350:100:50:0.5 containing 5% phosphoric acid after neutral phospholipids had been eluted with methanol. It was verified that extremely high recoveries of cholesterol (CH), triglycerides (TG), free fatty acids (FFA), phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), phosphatidic acid (PA), sphingomyelin (SM), and cerebrosides were obtained with this method. In addition, there appeared to be no preferential losses or degradation of any particular molecular species as the fatty acid distribution of bovine brain PS and the molecular species profile of plant PI were unaltered by the procedure. Depending on the tissue, this method may yield fractions containing pure lipid classes and/or simple mixtures of lipid classes of similar polarity. These fractions may then be more easily separated by thin-layer chromatography or high performance liquid chromatography for a complete lipid class analysis.


Factors affecting the Affordable Care Act Marketplace stand-alone pediatric dental plan premiums.
Children from lower income families have inadequate dental insurance coverage and poorer dental health in the United States. The Affordable Care Act (ACA) created Health Insurance Exchange Marketplaces to increase competition among health insurers and to provide low-income families with less costly health plans. The study examined Marketplace pediatric stand-alone dental plans (SADPs) and factors that affect their premiums. The data used were 2016 Federal-Facilitated and State-Partnership Marketplace pediatric SADP data. Ordinary least squares regressions were applied to estimate contributing factors' effects on SADP premiums. Great premium variation was found among low and high coverage level SADPs, respectively. Premiums of Health Maintenance Organization (HMO) SADPs were significantly less expensive than Preferred Provider Organization (PPO) SADPs. SADPs charged significantly higher premiums for more types of services covered. SADPs also charged higher premiums in states where there are larger proportions of low-income people who report poor dental health, more dentists per capita, or higher dentists' wages. The number of insurance companies offering pediatric SADPs in a Marketplace was negatively associated with premiums. The current Marketplace pediatric SADPs may have limited effects on increasing economically disadvantaged children's access to quality dental care. Marketplaces can promote competition among its pediatric dental insurers on providing lower-cost pediatric SADPs.


Capsaicin-induced corneal lesions in mice and the effects of chemical sympathectomy.
Effects of chemical sympathectomy on corneal changes induced in mice by a s.c. injection of capsaicin were investigated. Pretreatment with a s.c. injection of 6-hydroxydopamine (6-OHDA) on the 1st and 2nd postnatal day or on the 14th and 15th postnatal day led to a marked suppression of the capsaicin-induced corneal lesions. This suppressive effect also was evident in case of administration after capsaicin treatment. Intraventricular injection of 6-OHDA had a slight, transient effect. DSP4 [N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine], another potent substance used for sympathetic denervation, had a suppressive effect similar to that of 6-OHDA. The concentration of capsaicin in tissues of the head was unaltered with 6-OHDA. The content of substance P (SP) in the ocular anterior segments was decreased, dose-dependently, with capsaicin administration. Neonatal administration of 6-OHDA decreased the rate of capsaicin-induced reduction of SP. However, this effect of 6-OHDA was too slight to explain the suppression of the corneal lesions, as the intensity score of lesions with a large dose of capsaicin after 6-OHDA was lower than that with a small dose of capsaicin without 6-OHDA, whereas SP content in the former was still much lower than that in the latter. On the other hand, the content of norepinephrine in the ocular tissues was decreased in the presence of 6-OHDA but not capsaicin. These results suggest that the corneal changes induced by capsaicin are largely inhibited by a decreased activity in the peripheral sympathetic system.


Isolation of a candidate gene, CAB1, for cholesterol transport to mitochondria from the c-ERBB-2 amplicon by a modified cDNA selection method.
An improved cDNA selection method was established to isolate expressed genes efficiently from an amplified chromosome region in human cancer. Biotinylated yeast artificial chromosome DNA containing c-ERBB-2 was hybridized in solution with PCR-amplifiable cDNAs of an esophageal cancer cell line bearing the c-ERBB-2 amplification. After capturing the hybrids on avidin-coated magnetic beads, the cDNAs were amplified by PCR. Four new genes (A39, C51, CAB1, and GRB-7) coamplified with c-ERBB-2 were isolated from the enriched cDNA library. CAB1, GRB-7, and c-ERBB-2 were overexpressed in gastric and esophageal cancer cells in correspondence with the amplification. The deduced amino acid sequence of the CAB1 gene had significant homology to the recently discovered steroidogenic acute regulatory protein, StAR, which plays an essential role in cholesterol transport to mitochondria. It was established that multiple overexpressed genes are frequently present in a single amplicon.


Endovascular recanalization for symptomatic intracranial internal carotid and middle cerebral artery occlusion lasting longer than 72 h: Experience in a single center.
The objective of this study was to assess the safety, feasibility, and outcomes of endovascular recanalization for symptomatic intracranial internal carotid and middle cerebral artery occlusion lasting longer than 72 h. Thirty-nine consecutive patients with symptomatic occlusion of the anterior circulation and failure of medical therapy underwent endovascular recanalization and were included in this retrospective study. Patient characteristics, atherosclerotic risk factors, successful recanalization rates, and angiographic data were collected. Recanalization was successful in 37 cases (94.9%). The average residual stenosis immediately after intervention was 11.6 ± 4.3%. The patients who underwent balloon angioplasty alone had similar residual stenosis to those who also underwent stent placement (15.6 ± 7.3% vs. 9.0 ± 6.4%, P = 0.184). Intra- and perioperative complications occurred in three cases (7.69%). One patient (2.7%) developed severe in-stent restenosis with transient ischemic attack symptoms at 1-year follow-up. Endovascular recanalization is feasible for symptomatic occlusion of the anterior circulation lasting longer than 72 h. Recanalization provides a higher success rate when performed within 6 months of the qualifying event.


Test-retest and intrarater reliability of 2-dimensional ultrasound measurements of distance between rectus abdominis in women.
Single-group test-retest reliability study. To evaluate the test-retest intraobserver reliability of 2-dimensional ultrasound measurement of the distance between the rectus abdominis muscles, the interrectus distance (IRD). Diastasis recti is defined as the separation of the 2 rectus abdominis muscles, with a reported prevalence of between 30% and 70% in women during pregnancy and in the postpartum period. The condition is difficult to measure, and ultrasound imaging has been suggested as a useful method to quantify the diastasis. However, to date, no studies have investigated intratester or intertester reliability of ultrasound to measure the distance between the rectus abdominis muscles during rest and contraction. Ultrasound images from the rectus abdominis were recorded in 24 healthy female volunteers at rest and under 2 conditions of abdominal contraction: abdominal crunch and drawing-in exercises. The probe was positioned at 2 locations: below and above the umbilicus. A blinded investigator measured the IRD offline from 2 different ultrasound images collected on 2 different days (test-retest). Additionally, reanalyses of the same ultrasound images were done on 2 separate occasions (intra-image). Test-retest measurements of IRD demonstrated good to very good reliability, with intraclass correlation coefficient values between 0.74 and 0.90. The only exception was for IRD measured 2 cm below the umbilicus during the abdominal crunch exercise, which had an intraclass correlation coefficient of 0.50. For intratester reliability of the same images, the intraclass correlation coefficient values were all above 0.90. Ultrasound imaging is a reliable method for measuring the IRD at rest and during abdominal crunch and drawing-in exercises.


[Capacity analysis of health food choice by reference to consumers in two models of nutritional labeling; crossover study].
The aim of this study was to compare two models of nutrition labeling front-of-pack, in reference to the ability of consumers to choose a diet closer to nutritional recommendations. Randomized crossover design in 32 adults (18-65 years) of both sexes. Participants were randomly exposed to two experimental conditions using nutritional traffic light system (S-SN) or monochrome system (SM). Participants had to choose options from a closed menu for five days on the basis of the experimental front-of-pack labelling. For each meal, three food options with different nutritional compositions were given to the participants. The total energy and fat, saturated fat, sugar and salt of the chosen options were calculated. No significant differences at baseline sociodemographic and anthropometric characteristics were shown between individuals regardless of the experimental condition in which they started. The subjects tended to choose a diet with a lower, but not significant energy content of 23.0 ± 67.5 (P = 0.063) and a significantly lower sugar content of 3.5 ± 9.2 g, P < 0.001 and 0.6 ± 1 g, P < 0.003 for salt. Compared to the to the monochrome system, the multiple traffic-light system probably can help make food choices with less sugar and salt in a situation similar to the usual purchase in which there is a time limitation.


Isolation and characterization of NIH 3T3 cells expressing polyomavirus small T antigen.
The polyomavirus small T-antigen gene, together with the polyomavirus promoter, was inserted into a retrovirus vector pGV16 which contains the Moloney sarcoma virus long terminal repeat and neomycin resistance gene driven by the simian virus 40 promoter. This expression vector, pGVST, was packaged into retrovirus particles by transfection of psi 2 cells which harbor packaging-defective murine retrovirus genome. NIH 3T3 cells were infected by this replication-defective retrovirus containing pGVST. Of the 15 G418-resistant cell clones, 8 express small T antigen at various levels as revealed by immunoprecipitation. A cellular protein with an apparent molecular weight of about 32,000 coprecipitates with small T antigen. Immunofluorescent staining shows that small T antigen is mainly present in the nuclei. Morphologically, cells expressing small T antigen are indistinguishable from parental NIH 3T3 cells and have a microfilament pattern similar to that in parental NIH 3T3 cells. Cells expressing small T antigen form a flat monolayer but continue to grow beyond the saturation density observed for parental NIH 3T3 cells and eventually come off the culture plate as a result of overconfluency. There is some correlation between the level of expression of small T antigen and the growth rate of the cells. Small T-antigen-expressing cells form small colonies in soft agar. However, the proportion of cells which form these small colonies is rather small. A clone of these cells tested did not form tumors in nude mice within 3 months after inoculation of 10(6) cells per animal. Thus, present studies establish that the small T antigen of polyomavirus is a second nucleus-localized transforming gene product of the virus (the first one being large T antigen) and by itself has a function which is to stimulate the growth of NIH 3T3 cells beyond their saturation density in monolayer culture.


Genetic analysis of slipper/mixed lineage kinase reveals requirements in multiple Jun-N-terminal kinase-dependent morphogenetic events during Drosophila development.
Mixed lineage kinases (MLKs) function as Jun-N-terminal kinase (JNK) kinase kinases to transduce extracellular signals during development and homeostasis in adults. slipper (slpr), which encodes the Drosophila homolog of mammalian MLKs, has previously been implicated in activation of the JNK pathway during embryonic dorsal epidermal closure. To further define the specific functions of SLPR, we analyzed the phenotypic consequences of slpr loss and gain of function throughout development, using a semiviable maternal-effect allele and wild-type or dominant-negative transgenes. From these analyses we confirm that failure of dorsal closure is the null phenotype in slpr germline clones. In addition, there is a functional maternal contribution, which can suffice for embryogenesis in the zygotic null mutant, but rarely suffices for pupal metamorphosis, revealing later functions for slpr as the maternal contribution is depleted. Zygotic null mutants that eclose as adults display an array of morphological defects, many of which are shared by hep mutant animals, deficient in the JNK kinase (JNKK/MKK7) substrate for SLPR, suggesting that the defects observed in slpr mutants primarily reflect loss of hep-dependent JNK activation. Consistent with this, the maternal slpr contribution is sensitive to the dosage of positive and negative JNK pathway regulators, which attenuate or potentiate SLPR-dependent signaling in development. Although SLPR and TAK1, another JNKKK family member, are differentially used in dorsal closure and TNF/Eiger-stimulated apoptosis, respectively, a Tak1 mutant shows dominant genetic interactions with slpr, suggesting potential redundant or combinatorial functions. Finally, we demonstrate that SLPR overexpression can induce ectopic JNK signaling and that the SLPR protein is enriched at the epithelial cell cortex.


Dependence of peptidoglycan metabolism on phospholipid synthesis during growth of Escherichia coli.
The role of phospholipid synthesis in peptidoglycan metabolism during growth of Escherichia coli was determined. The inhibition of phospholipid synthesis, achieved by inhibiting fatty acid synthesis with cerulenin or by glycerol deprivation of gpsA mutant strains, resulted in the concomitant inhibition of peptidoglycan synthesis. These effects on peptidoglycan synthesis were relatively specific in that the treatments did not cause a general inhibition of macromolecular synthesis. Furthermore, the inhibition of phospholipid synthesis also resulted in the rapid development of penicillin tolerance. It was unlikely that penicillin tolerance in these cases were simply due to the inhibition of growth caused by cerulenin treatment or glycerol deprivation because treatments with more effective growth inhibitors, e.g. chloramphenicol or norfloxacin, did not confer penicillin tolerance. Penicillin tolerance was shown to be a direct consequence of the inhibition of phospholipid synthesis and not due to the possible accumulation of guanosine-3',5'-bispyrophosphate (ppGpp), the starvation stress signal molecule known to be responsible for the development of penicillin tolerance in amino-acid-deprived bacteria. Therefore, peptidoglycan metabolism is coupled to phospholipid synthesis during growth of E. coli, and this may represent an important means to ensure the coordination of cell envelope synthesis in growing bacteria.


The evolution of birdsong on islands.
Islands are simplified, isolated ecosystems, providing an ideal set-up to study evolution. Among several traits that are expected to change on islands, an interesting but poorly understood example concerns signals used in animal communication. Islands are typified by reduced species diversity, increased population density, and reduced mate competition, all of which could affect communication signals. We used birdsong to investigate whether there are systematic changes in communication signals on islands, by undertaking a broad comparison based on pairs of closely related island-mainland species across the globe. We studied song traits related to complexity (number of different syllables, frequency bandwidth), to vocal performance (syllable delivery rate, song duration), and also three particular song elements (rattles, buzzes, and trills) generally implicated in aggressive communication. We also investigated whether song complexity was related to the number of similar sympatric species. We found that island species were less likely to produce broadband and likely aggressive song elements (rattles and buzzes). By contrast, various aspects of song complexity and performance did not differ between island and mainland species. Species with fewer same-family sympatric species used wider frequency bandwidths, as predicted by the character release hypothesis, both on continents and on islands. Our study supports the hypothesis of a reduction in aggressive behavior on islands and suggests that discrimination against closely related species is an important factor influencing birdsong evolution.


Sequences of herpes simplex virus type 1 that inhibit formation of stable TK+ transformants.
We have identified two regions of the herpes simplex virus type 1 (HSV-1) genome that inhibit DNA-mediated transformation of thymidine kinase-less L (Ltk-) cells by the cloned HSV-1 tk gene. When plasmids containing the EcoRI fragments EK or JK were mixed at 30 fmol/ml with the tk gene and transfected into Ltk- cells, the frequency of transformation was inhibited 80 to more than 90% relative to the control. Of the remaining 10 EcoRI fragments of the HSV-1 genome, 8 were inactive and 2 were weakly active. A 6.1-kilobase PstI subclone between 0.743 and 0.782 map units was isolated from pEK. This clone, pEK-P3P4, exhibited antitransformation activity toward HSV-1 tk and also the bacterial genes gpt and neo. pEK-P3P4 contains the alpha 27 gene, and restriction endonuclease inactivation and subcloning studies established that alpha 27 alone did not inhibit transformation. However, alpha 27 plus sequences both upstream and downstream of alpha 27 did inhibit transformation. In addition, alpha 0 or alpha 4 could substitute for alpha 27 in effecting antitransformation with these sequences. Therefore, an alpha gene and two additional loci in pEK-P3P4 are required for antitransformation. A second antitransforming locus in the reiterated sequences common to EK and JK and distinct from those in pEK-P3P4 was also identified but not characterized in detail. How antitransformation may be an expression of regulation of viral and host cell gene expression is discussed.


PNA microprobe for label-free detection of expanded trinucleotide repeats.
We present a PNA microprobe sensing platform to detect trinucleotide repeat mutation by electrochemical impedance spectroscopy. The microprobe platform discriminated Huntington's disease-associated CAG repeats in cell-derived total RNA with S/N 1 : 3. This sensitive, label-free, and PCR-free detection strategy may be employed in the future to develop biosensing platforms for the detection of a plethora of repeat expansion disorders.


A systematic review of ankle fracture treatment modalities in diabetic patients.
This systematic review evaluated the surgical outcomes of various ankle fracture treatment modalities in patients with Diabetes Mellitus as well as the methodological quality of the studies. For our review, four online databases were searched: PubMed, MEDLINE (Clarivate Analytics), CINAHL (Cumulative Index to Nursing and Allied Health) and Web of Science (Clarivate Analytics). The overall methodological quality of the studies was assessed with the Coleman Methodology Score. Data regarding diabetic ankle fractures were pooled into three outcomes groups for comparison: (1) the standard fixation cohort with management of diabetic ankle fractures using ORIF with small or mini fragment internal fixation techniques following AO principles, (2) the minimally invasive cohort with diabetic ankle fracture management utilizing percutaneous cannulated screws or intramedullary fixation, and (3) the combined construct cohort treated with a combination of ORIF and another construct (transarticular or external fixation). The search strategy identified 2228 potential studies from the four databases and 11 were included in the final review. Compared to the standard fixation cohort, the minimally invasive cohort had increased risk of hardware breakage or migration and the combined constructs cohort had increased risk of hardware breakage or migration, surgical site infection and nonunion. Limb salvage rates were similar for the standard fixation and minimally invasive cohorts; however, the combined constructs cohort had a significantly lower limb salvage rate compared to that of the standard fixation cohort. The mean Coleman Methodology Score indicated the quality of the studies in the review was poor and consistent with its limitations. The overall quality of published studies on operative treatment of diabetic ankle fractures is low. Treating diabetic ankle fractures operatively results in a high number of complications regardless of fixation method. However, limb salvage rates remain high overall at 97.9% at a mean follow-up of 21.7 months. To achieve improved limb salvage rates and decrease complications, it is critical is to follow basic AO principles, respect the soft tissue envelope or utilize minimally invasive techniques, and be wary that certain combined constructs may be associated with higher complication rates. 2.


Visceral larva migrans. A review and reassessment indicating two forms of clinical expression: visceral and ocular.
Visceral larva migrans is a disease in which the larvae of animal parasites invade human tissues but do not complete their life cycles. The most frequent cause of this illness in children is the dog roundworm, Toxocara canis. A review of the literature, as well as our clinical experience, indicates that there are two forms of clinical expression: one, visceral, and the other, ocular. In general the clinical and laboratory abnormalities (hepatomegaly, recurrent pneumonia, eosinophilia, and hyperglobulinemia) usually associated with visceral disease are absent in children with ocular abnormalities. Conversely, there is a general lack of eye complications in patients with systemic disease. Reasons for these variations in clinical expression are unknown, but immune responses of the host and the antigenic composition of the parasite may be contributing factors.


[Retroperitoneal haematoma following groin injury].
Retroperitoneal haematoma, a rare complication in patients with traumatism or surgery of the inguinal region, may be clinically silent and difficult diagnosis. A 14-year-old man with retroperitoneal haematoma following groin traumatism is reported. Authors analyse the anatomy of the inguinal region and their relationship with retroperitoneal space concluding that clinical suspicion and computed tomography are the elements of choice in the diagnosis of this complication.


Extraction of correlated gene clusters from multiple genomic data by generalized kernel canonical correlation analysis.
A major issue in computational biology is the reconstruction of pathways from several genomic datasets, such as expression data, protein interaction data and phylogenetic profiles. As a first step toward this goal, it is important to investigate the amount of correlation which exists between these data. These methods are successfully tested on their ability to recognize operons in the Escherichia coli genome, from the comparison of three datasets corresponding to functional relationships between genes in metabolic pathways, geometrical relationships along the chromosome, and co-expression relationships as observed by gene expression data.


AMC Datasets-a resource for analytical scientists.
Have you noticed, when downloading a Technical Brief from the AMC's web pages (www.rsc.org/amc), that there is a section called AMC Datasets listed in the leftmost column? This content was inaugurated some years ago to provide a permanent collection of interesting datasets related to analytical chemistry and its applications. The basic idea was to provide analytical chemists with material that could be used to support teaching, learning and research in statistics and chemometrics. New ideas in these fields could be tested on real and well-characterised datasets, and compared with results of other workers.