In Vitro Differentiation of Neural-Like Cells from Human Embryonic Stem Cells by A Combination of Dorsomorphin, XAV939, and A8301.
Motor neuron differentiation from human embryonic stem cells (hESCs) is a goal of regenerative medicine to provide cell therapy as treatments for diseases that damage motor neurons. Most protocols lack adequate efficiency in generating functional motor neurons. However, small molecules present a new approach to overcome this challenge. The aim of this research is to replace morphogen factors with a cocktail of efficient, affordable small molecules for effective, low cost motor neuron differentiation. In this experimental study, hESCs were differentiated into motor neuron by the application of a small molecule cocktail that consisted of dorsomorphin, A8301, and XAV939. During the differentiation protocol, we selected five stages and assessed expressions of neural markers by real-time polymerase chain reaction (PCR), immunofluorescence staining, and flow cytometry. Motor neuron ion currents were determined by whole cell patch clamp recording. Immunofluorescence staining and flow cytometry analysis of hESC-derived neural ectoderm (NE) indicated that they were positive for NESTIN (92.68%), PAX6 (64.40%), and SOX1 (82.11%) in a chemically defined adherent culture. The replated (hESC)-derived NE differentiated cells were positive for TUJ1, MAP2, HB9 and ISL1. We evaluated the gene expression levels with real-time reverse transcriptase-PCR at different stages of the differentiation protocol. Voltage gated channel currents of differentiated cells were examined by the whole-cell patch clamp technique. The hESC-derived motor neurons showed voltage gated delay rectifier K+, Na+ and Ca2+ inward currents. Our results indicated that hESC-derived neurons expressed the specific motor neuron markers specially HB9 and ISL1 but voltage clamp recording showed small ionic currents therefore it seems that voltage gated channel population were inadequate for firing action potentials.


Exercise-induced injury to skeletal muscle.
Strenuous or unaccustomed exercise can cause injury to skeletal muscle. This paper reviews our understanding of the mechanisms of exercise-induced injury. Measurements of exercise-induced injury have included muscle soreness, increased serum levels of intracellular enzymes, increased lysosomal enzyme activities, structural changes in muscle fibers, and prolonged decreases in force development that cannot be attributed to fatigue. Injury can be induced by exercise of small muscle groups, which suggests that it involves processes localized in skeletal muscles. Exercise of relatively short duration can result in injury, which indicates that long durations of exercise and associated metabolic changes are not necessary for injury to occur. Exercise that involves lengthening contractions results in greater evidence of muscle injury than exercise involving isometric or shortening contractions. Lengthening contractions are associated with higher levels of force and lower metabolic costs per muscle fiber than isometric or shortening contractions. These results suggest that changes in muscle metabolism are not responsible for exercise-induced injury to skeletal muscle. Exercise-induced injury is more likely the result of mechanical disruption of muscle fibers.


A Maize Male Gametophyte-Specific Gene Encodes ZmLARP6c1, a Potential RNA-Binding Protein Required for Competitive Pollen Tube Growth.
Members of the La-related protein family (LARPs) contain a conserved La module, which has been associated with RNA-binding activity. Expression of the maize gene GRMZM2G323499/Zm00001d018613, a member of the LARP family, is highly specific to pollen, based on both transcriptomic and proteomic assays. This suggests a pollen-specific RNA regulatory function for the protein, designated ZmLARP6c1 based on sequence similarity to the LARP6 subfamily in Arabidopsis. To test this hypothesis, a Ds-GFP transposable element insertion in the ZmLarp6c1 gene (tdsgR82C05) was obtained from the Dooner/Du mutant collection. Sequencing confirmed that the Ds-GFP insertion is in an exon, and thus likely interferes with ZmLARP6c1 function. Tracking inheritance of the insertion via its endosperm-expressed GFP indicated that the mutation was associated with reduced transmission from a heterozygous plant when crossed as a male (ranging from 0.5 to 26.5% transmission), but not as a female. Furthermore, this transmission defect was significantly alleviated when less pollen was applied to the silk, reducing competition between mutant and wild-type pollen. Pollen grain diameter measurements and nuclei counts showed no significant differences between wild-type and mutant pollen. However, in vitro, mutant pollen tubes were significantly shorter than those from sibling wild-type plants, and also displayed altered germination dynamics. These results are consistent with the idea that ZmLARP6c1 provides an important regulatory function during the highly competitive progamic phase of male gametophyte development following arrival of the pollen grain on the silk. The conditional, competitive nature of the Zmlarp6c1::Ds male sterility phenotype (i.e., reduced ability to produce progeny seed) points toward new possibilities for genetic control of parentage in crop production.


Detection of immunoperoxidase labelled mycoplasmas in cell culture by light microscopy and electronmicroscopy.
A McCoy cell line persistently infected with Mycoplasma orale was examined by light microscopy and electronmicroscopy after specific labelling with a direct immunoperoxidase conjugate. Mycoplasmas were readily detected in monolayer cultures in bright-field conditions and these were related to labelled organisms observed by electronmicroscopy in thin sections of similar cells. The specificity of the conjugate for M. orale was demonstrated by blocking with unconjugated antiserum, and by its inability to detect M. bovirhinis. Non-specific background labelling was consistently absent.


Adenovirus-mediated transfer of p33ING1 with p53 drastically augments apoptosis in gliomas.
The p53 tumor suppressor gene is an important target for the gene therapy of cancers, and clinical trials targeting this gene have been conducted. Some cancers, however, are refractory to p53 gene therapy. Therefore, it has been combined with other therapies, including chemotherapy and radiotherapy, to enhance the cytopathic effect of p53 induction. The p33ING1 gene cooperates with p53 to block cell proliferation. In this study, we investigated whether adenovirus (Adv)-mediated coinduction of p33ING1 and p53 enhances apoptosis in glioma cells (U251 and U-373 MG), which showed no genetic alterations but low expression levels of p33ING1. Although the single infection of Adv for p33ING1 (Adv-p33) at a multiplicity of infection (MOI) of 100, or Adv for p53 controlled by myelin basic protein (MBP) promoter (Adv-MBP-p53), a glioma-specific promoter, at a MOI of 50, did not induce apoptosis in U251 and U-373 MG glioma cells; coinfection of Adv-p33 and Adv-MBP-p53 at the same MOIs induced drastically enhanced apoptosis in both cell lines. Apoptosis was not induced in NGF-treated PC-12 cells infected with a high MOI (300) of Adv-p33 nor in those coinfected with Adv-p33 (100) and Adv-MBP-p53 (50). Coinfection of Adv-p33 and Adv-MBP-p53 demonstrated morphological mitochondrial damage during the initial stage of apoptosis, which likely led to apoptotic cell death. Our results indicate that this coinfection approach can be used as a modality for the gene therapy of gliomas, sparing damage to normal tissues.


Nucleolin mediates nucleosome disruption critical for DNA double-strand break repair.
Recruitment of DNA repair factors and modulation of chromatin structure at sites of DNA double-strand breaks (DSBs) is a complex and highly orchestrated process. We developed a system that can induce DSBs rapidly at defined endogenous sites in mammalian genomes and enables direct assessment of repair and monitoring of protein recruitment, egress, and modification at DSBs. The tight regulation of the system also permits assessments of relative kinetics and dependencies of events associated with cellular responses to DNA breakage. Distinct advantages of this system over focus formation/disappearance assays for assessing DSB repair are demonstrated. Using ChIP, we found that nucleosomes are partially disassembled around DSBs during nonhomologous end-joining repair in G1-arrested mammalian cells, characterized by a transient loss of the H2A/H2B histone dimer. Nucleolin, a protein with histone chaperone activity, interacts with RAD50 via its arginine-glycine rich domain and is recruited to DSBs rapidly in an MRE11-NBS1-RAD50 complex-dependent manner. Down-regulation of nucleolin abrogates the nucleosome disruption, the recruitment of repair factors, and the repair of the DSB, demonstrating the functional importance of nucleosome disruption in DSB repair and identifying a chromatin-remodeling protein required for the process. Interestingly, the nucleosome disruption that occurs during DSB repair in cycling cells differs in that both H2A/H2B and H3/H4 histone dimers are removed. This complete nucleosome disruption is also dependent on nucleolin and is required for recruitment of replication protein A to DSBs, a marker of DSB processing that is a requisite for homologous recombination repair.


Assessment of global reporting of adverse drug reactions for anti-malarials, including artemisinin-based combination therapy, to the WHO Programme for International Drug Monitoring.
In spite of enhanced control efforts, malaria remains a major public health problem causing close to a million deaths annually. With support from several donors, large amounts of artemisinin-based combination therapy (ACT) are being deployed in endemic countries raising safety concerns as little is known about the use of ACT in several of the settings where they are deployed. This project was undertaken to profile the provenance of the pharmacovigilance reporting of all anti-malarials, including ACT to the WHO adverse drug reaction (ADR) database (Vigibase™) over the past 40 years. The WHO Programme for International Drug Monitoring, the Uppsala Monitoring Centre (UMC) provided anonymized extracts of Vigibase™ covering the period 1968-2008. All countries in the programme were clustered according to their malaria control phase and income status. The number of individual case safety reports (ICSRs) of anti-malarials was analyzed according to those clusters. From 1968 to 2008, 21,312 ICSRs suspecting anti-malarials were received from 64 countries. Low-income countries, that are also malaria-endemic (categorized as priority 1 countries) submitted only 1.2% of the ICSRs. Only 60 out of 21,312 ICSRs were related to ACT, 51 of which were coming from four sub-Saharan African countries. Although very few ICSRs involved artemisinin-based compounds, many of the adverse events reported were potentially serious. This paper illustrates the low reporting of ADRs to anti-malarials in general and ACT in particular. Most reports were submitted by non-endemic and/or high-income countries. Given the current mix of large donor funding, the insufficient information on safety of these drugs, increasing availability of ACT and artemisinin-based monotherapies in public and private sector channels, associated potential for inappropriate use and finally a pipeline of more than 10 new novel anti-malarials in various stages of development, the presence of well functioning national pharmacovigilance systems is vital to ensure safe and responsible scale up of ACT deployment. Bringing together the competencies of national pharmacovigilance centres and various types of organizations in the NGO, academic and private sectors with global coordination to create short- and long-term solutions may help address the lag between rapidly growing ACT use and poor ADR reporting.


Antioxidant Effect of Lonicera caerulea L. in the Cardiovascular System of Obese Zucker Rats.
Lonicera caerulea L. (Loni) represents a promising source of beneficial polyphenols with therapeutical potential in cardiovascular diseases. We aimed to study the effects of Loni and coenzyme Q10 (CoQ10) on selected cardiometabolic parameters and NO/ROS balance in obese Zucker rats. Male Zucker rats were divided into the control group and groups treated with CoQ10 (30 mg/kg/day) or Loni (5 g/kg/day) for 6 weeks. Blood pressure, body weight, heart weight, and plasma lipid profile were determined. NOS activity and protein expressions of eNOS, SOD, NADPH oxidase, and NF-kappa B were measured in the heart and aorta. Neither body weight nor blood pressure were significantly changed after six weeks of Loni or CoQ10 treatment. Both Loni and CoQ10 decreased the plasma LDL level. Moreover, Loni decreased the total cholesterol level. The total NOS activity did not change in the heart after the treatments. However, in the aorta, Loni treatment increased NOS activity and protein expression of SOD and decreased expressions of NADPH oxidase and NF-kappa B compared to both the control and CoQ10 groups. There were no changes in the eNOS protein expression within the groups. In conclusion, it seems that the antioxidant effect of Loni was responsible for both the decrease of plasma LDL and the total cholesterol levels and the increase of vascular NOS activity.


Biochemical Evaluation of the Effects of Concentrated Growth Factor Liquid on Osseointegration: A Split-Mouth Design Study.
To investigate cytokine levels in peri-implant crevicular fluid and thus evaluate the effects of concentrated growth factor (CGF) on osseointegration. A total of 40 mandibular implants were symmetrically placed in a group of 20 systemically healthy patients enrolled in the study. In each patient, one implant wetted with liquid infiltrated from fibrin matrix was placed in the test side (Group L), and the other implant was placed in the control side without the application of any material (Group C). Peri-implant crevicular fluid was collected at 2, 4, and 12 weeks later. Marginal bone loss was measured with panoramic radiographs taken immediately after implant placement and at 12 weeks. Resonance frequency analysis (RFA) of the implants was performed intraoperatively and at 4 and 12 weeks. Stability values of the implants in the CGF liquid-treated sites were higher than those of the control group at week 12 (P = .005). There was no statistically significant difference between the two groups in terms of marginal bone loss (MBL). Group L showed increased levels of tumor necrosis factor alpha (TNF-α) and receptor activator of nuclear factor kappa-B ligand (RANKL) at 2 and 4 weeks. Also, levels of osteoprotegerin (OPG) were higher in Group L at week 4 compared to Group C (P = .033). The increased TNF-α, RANKL, and OPG levels in this study demonstrate that CGF liquid can be used to accelerate peri-implant bone remodeling in the early phase of osseointegration.


The Division of Behavioral and Social Research (BSR) at the National Institute on Aging (NIA), National Institutes of Health (NIH), is the leading U.S. funder of social, behavioral, psychological, and economic research on the processes of aging at the individual and population level. A cornerstone of our investments is the Health and Retirement Study (HRS), a longitudinal study launched by NIA in 1992 with support from the Social Security Administration.


PROMs data: can it be used to make decisions for individual patients? A narrative review.
Patient-reported outcome measures (PROMs) are increasingly used in clinical practice providing health care professionals with patients' perceptions and views of their health. They have traditionally been utilized in health research and health service evaluation and are now starting to be used in routine clinical practice with individual patients. The repeated administration of PROMs over the course of care with individual patients has a role in patient assessment, assisting clinical decision-making, and tracking patient progress. This approach can influence the patient-clinician encounter impacting the therapeutic alliance and increasing patient engagement with care. It is also theorized to improve patient outcomes and satisfaction with care. Advances in technologies and innovations in methodology have led to the use of electronic systems to simplify the collection and reporting of PROMs. Challenges of using PROMs with individual patients include clinician knowledge and skills, and access to appropriate technology. This paper reviews the use of PROMs with individual patients, illustrating how they may affect the patient-clinician encounter impact satisfaction and health outcomes. The routine use of PROMs during a course of care rather than just at the start and end provides additional opportunity to inform clinician and patient with benefits to both. The adoption of PROMs in clinical practice can help health care professionals to make decisions for individual patients. Further work is needed to examine the implementation of PROMs and benefits of PROMs in different clinical contexts.


Apical F-actin-regulated exocytic targeting of NtPPME1 is essential for construction and rigidity of the pollen tube cell wall.
In tip-confined growing pollen tubes, delivery of newly synthesized cell wall materials to the rapidly expanding apical surface requires spatial organization and temporal regulation of the apical F-actin filament and exocytosis. In this study, we demonstrate that apical F-actin is essential for the rigidity and construction of the pollen tube cell wall by regulating exocytosis of Nicotiana tabacum pectin methylesterase (NtPPME1). Wortmannin disrupts the spatial organization of apical F-actin in the pollen tube tip and inhibits polar targeting of NtPPME1, which subsequently alters the rigidity and pectic composition of the pollen tube cell wall, finally causing growth arrest of the pollen tube. In addition to mechanistically linking cell wall construction and apical F-actin, wortmannin can be used as a useful tool for studying endomembrane trafficking and cytoskeletal organization in pollen tubes.


A new approach towards biomarker selection in estimation of human exposure to chiral chemicals: a case study of mephedrone.
Wastewater-based epidemiology is an innovative approach to estimate public health status using biomarker analysis in wastewater. A new compound detected in wastewater can be a potential biomarker of an emerging trend in public health. However, it is currently difficult to select new biomarkers mainly due to limited human metabolism data. This manuscript presents a new framework, which enables the identification and selection of new biomarkers of human exposure to drugs with scarce or unknown human metabolism data. Mephedrone was targeted to elucidate the assessment of biomarkers for emerging drugs of abuse using a four-step analytical procedure. This framework consists of: (i) identification of possible metabolic biomarkers present in wastewater using an in-vivo study; (ii) verification of chiral signature of the target compound; (iii) confirmation of human metabolic residues in in-vivo/vitro studies and (iv) verification of stability of biomarkers in wastewater. Mephedrone was selected as a suitable biomarker due to its high stability profile in wastewater. Its enantiomeric profiling was studied for the first time in biological and environmental matrices, showing stereoselective metabolism of mephedrone in humans. Further biomarker candidates were also proposed for future investigation: 4'-carboxy-mephedrone, 4'-carboxy-normephedrone, 1-dihydro-mephedrone, 1-dihydro-normephedrone and 4'-hydroxy-normephedrone.


Computer assisted photo-anthropometric analyses of full-face and profile facial images.
Expert witnesses using facial comparison techniques are regularly required to disambiguate cases of disputed identification in CCTV images and other photographic evidence in court. This paper describes a novel software-assisted photo-anthropometric facial landmark identification system, DigitalFace tested against a database of 70 full-face and profile images of young males meeting a similar description. The system produces 37 linear and 25 angular measurements across the two viewpoints. A series of 64 analyses were conducted to examine whether separate novel probe facial images of target individuals whose face dimensions were already stored within the database would be correctly identified as the same person. Identification verification was found to be unreliable unless multiple distance and angular measurements from both profile and full-face images were included in an analysis.


Reduction and expansion in microsporidian genome evolution: new insights from comparative genomics.
Microsporidia are an abundant group of obligate intracellular parasites of other eukaryotes, including immunocompromised humans, but the molecular basis of their intracellular lifestyle and pathobiology are poorly understood. New genomes from a taxonomically broad range of microsporidians, complemented by published expression data, provide an opportunity for comparative analyses to identify conserved and lineage-specific patterns of microsporidian genome evolution that have underpinned this success. In this study, we infer that a dramatic bottleneck in the last common microsporidian ancestor (LCMA) left a small conserved core of genes that was subsequently embellished by gene family expansion driven by gene acquisition in different lineages. Novel expressed protein families represent a substantial fraction of sequenced microsporidian genomes and are significantly enriched for signals consistent with secretion or membrane location. Further evidence of selection is inferred from the gain and reciprocal loss of functional domains between paralogous genes, for example, affecting transport proteins. Gene expansions among transporter families preferentially affect those that are located on the plasma membrane of model organisms, consistent with recruitment to plug conserved gaps in microsporidian biosynthesis and metabolism. Core microsporidian genes shared with other eukaryotes are enriched in orthologs that, in yeast, are highly expressed, highly connected, and often essential, consistent with strong negative selection against further reduction of the conserved gene set since the LCMA. Our study reveals that microsporidian genome evolution is a highly dynamic process that has balanced constraint, reductive evolution, and genome expansion during adaptation to an extraordinarily successful obligate intracellular lifestyle.


Degradation of T-Tubular Microdomains and Altered cAMP Compartmentation Lead to Emergence of Arrhythmogenic Triggers in Heart Failure Myocytes: An in silico Study.
Heart failure (HF) is one of the most common causes of morbidity and mortality worldwide. Although many patients suffering from HF die from sudden cardiac death caused by arrhythmias, the mechanism linking HF remodeling to an increased arrhythmogenic propensity remains incomplete. HF is typically characterized by a progressive loss of transverse tubule (T-tubule) domains, which leads to an altered distribution of L-type calcium channels (LTCCs). Microdomain degradation also causes the disruption of the β2 adrenergic receptor (β2AR) and phosphodiesterase (PDE) signaling localization, normally confined to the dyadic space. The goal of this study was to analyze how these subcellular changes affect the function of LTCCs and lead to the emergence of ventricular cell-level triggers of arrhythmias. To accomplish this, we developed a novel computational model of a human ventricular HF myocyte in which LTCCs were divided into six different populations, based on their location and signaling environment they experience. To do so, we included T-tubular microdomain remodeling which led to a subset of LTCCs to be redistributed from the T-tubular to the surface membrane and allowed for different levels of phosphorylation of LTCCs by PKA, based on the presence of β2ARs and PDEs. The model was used to study the behavior of the LTCC current (ICaL) under basal and sympathetic stimulation and its effect on cellular action potential. Our results showed that channels redistributed from the T-tubular membrane to the bulk of the sarcolemma displayed an altered function in their new, non-native signaling domain. Incomplete calcium dependent inactivation, which resulted in a longer-lasting and larger-in-magnitude LTCC current, was observed when we decoupled LTCCs from ryanodine receptors and removed them from the dyadic space. The magnitude of the LTCC current, especially in the surface sarcolemma, was also increased via phosphorylation by the redistributed β2ARs and PDEs. These changes in LTCC current led to the development of early afterdepolarizations. Thus, our study shows that altered LTCC function is a potential cause for the emergence of cell-level triggers of arrhythmia, and that β2ARs and PDEs present useful therapeutic targets for treatment of HF and prevention of sudden cardiac death.


Worldwide Assessment of the Root and Root Canal Characteristics of Maxillary Premolars - A Multi-center Cone-beam Computed Tomography Cross-sectional Study With Meta-analysis.
This worldwide study examined the root and root canal characteristics of maxillary premolars and explored how demographic factors impact the outcomes. Observers from 44 countries assessed 26,400 maxillary premolars using cone-beam computed tomography and employed a standardized screening method to gather data on multiple canal morphology and 3-rooted configuration (primary outcomes), as well as secondary outcomes related to root and root canal anatomies. Demographic factors such as ethnicity, sex, and age were collected for each participant. The intra- and inter-observer tests ensured observer reliability. Primary outcomes were represented as odds ratios and untransformed proportions accompanied by 95% confidence interval (CI) forest plots. Meta-analysis compared sub-groups and identified sources of heterogeneity (α = 5%). The overall prevalence of multiple canal morphology in the first premolar was 93.5% (95% CI, 93.3%-94.7%), while in the second premolar, it was 49.7% (95% CI, 44.9%-54.6%). The proportion of 3-rooted configuration was 1.8% (95% CI, 1.4%-2.1%) in the first premolar and 0.4% (95% CI, 0.3%-0.5%) in the second. Asian countries generally displayed fewer roots and root canals, while European nations showed higher counts (P < .05). Males exhibited higher percentages and odds ratios for both outcomes in both premolars. Younger patients demonstrated lower percentages of multiple root canal morphologies (P < .05). Factors such as tooth side, voxel size and field-of-view did not influence the outcomes (P > .05). The worldwide assessment of root and root canal characteristics of maxillary premolars has revealed a discernible influence of various factors such as tooth type, geographical region, ethnicity, sex, and age.


Fenofibrate improves endothelial function and decreases thrombin-activatable fibrinolysis inhibitor concentration in metabolic syndrome.
Procoagulant state, inflammation, and endothelial dysfunction have been documented in metabolic syndrome. Endothelial dysfunction is a strong predictor of cardiovascular events. Studies on the association of thrombin-activatable fibrinolysis inhibitor and thrombosis are still controversial, but substantial evidence suggests that increased thrombin-activatable fibrinolysis inhibitor or thrombin-activatable fibrinolysis inhibits or protects against arterial thrombosis. This study aimed to assess concomitantly the effects of fenofibrate therapy on thrombin-activatable fibrinolysis inhibitor concentrations and endothelial functions in patients with metabolic syndrome. Twenty-five patients (16 women; mean age 50.4 +/- 7.0) were enrolled in the study. Plasma thrombin-activatable fibrinolysis inhibitor, C-reactive protein, and fibrinogen levels were measured before fenofibrate administration and after 8 weeks of fenofibrate treatment. Endothelial function was assessed by endothelial-dependent flow-mediated dilatation from brachial artery. Pretreatment (baseline) thrombin-activatable fibrinolysis inhibitor level was 52.3 (1.2-119.7) decreasing to 7.7 (0.9-51.2; P < 0.001) after 8 weeks of fibrate treatment. Endothelial functions, which were measured with flow-mediated dilatation, were significantly improved after treatment (mean flow-mediated dilatation was 6.76 +/- 2.21 at baseline and 10.66 +/- 1.17% after 8 week of fenofibrate treatment, P < 0.001). Fenofibrate decreases thrombin-activatable fibrinolysis inhibitor levels and improves endothelial function in metabolic syndrome and, thus, suggests a potential for protection against cardiovascular effects. Further studies are warranted to confirm the effects of fibrates on thrombin-activatable fibrinolysis inhibitor and for conclusive evidence on the association between thrombin-activatable fibrinolysis inhibitor and thrombosis.


Arguments used by proponents and opponents in Brazil's regulatory discussions of e-cigarettes and heated tobacco products.
To identify proponents and opponents of the commercialisation and marketing of e-cigarettes and heated tobacco products (HTPs), identify the arguments used on both sides and compare how the arguments have changed over time, we analysed three policy discussions occurring in 2009, 2018 and 2019. We conducted a content analysis of one document and six videos from these discussions, provided on the Brazilian Health Regulatory Agency website, or upon request. The arguments most used by tobacco companies were related to claims that the use of e-cigarettes and HTPs is less harmful than conventional tobacco. Unions that support its commercialisation also argued that lifting the ban would prevent smuggling and guarantee their quality. On the other side, universities, medical and anti-tobacco institutions argued that such devices may have health risks, including the risk of inducing cigarette smoking. In 2009, most arguments belonged to the 'health' theme, while in 2018 and 2019 economic arguments and those related to morals and ethics were frequently used. Those that supported the commercialisation and marketing of e-cigarettes and HTPs first focused on arguments of harm reduction, while 10 years later the right to access and potential economic consequences also became common. Public health agents and academics must gather evidence to effectively respond to these arguments and discuss these policies, and must prepare themselves to use and respond to arguments related to moral and economic themes.


Privacy-preserving clinical decision support system using Gaussian kernel-based classification.
A clinical decision support system forms a critical capability to link health observations with health knowledge to influence choices by clinicians for improved healthcare. Recent trends toward remote outsourcing can be exploited to provide efficient and accurate clinical decision support in healthcare. In this scenario, clinicians can use the health knowledge located in remote servers via the Internet to diagnose their patients. However, the fact that these servers are third party and therefore potentially not fully trusted raises possible privacy concerns. In this paper, we propose a novel privacy-preserving protocol for a clinical decision support system where the patients' data always remain in an encrypted form during the diagnosis process. Hence, the server involved in the diagnosis process is not able to learn any extra knowledge about the patient's data and results. Our experimental results on popular medical datasets from UCI-database demonstrate that the accuracy of the proposed protocol is up to 97.21% and the privacy of patient data is not compromised.


Vascular function and multiple sclerosis.
Multiple sclerosis (MS) is a debilitating disease with an assumed autoimmune etiology which may lead to elevated oxidative stress, vascular dysfunction, and subsequent predisposition to cardiovascular disease. Therefore, the primary aim of this study was to evaluate vascular function and the potential role of oxidative stress in patients diagnosed with MS compared to healthy controls (C). Fourteen patients with relapsing-remitting MS (47 ± 3 years) and 13 age- and activity-matched controls (44 ± 5 years) underwent brachial artery flow-mediated dilation (FMD) and reactive hyperemia testing using ultrasound Doppler. Venous blood was analyzed for C-reactive protein (CRP), lipid hydroperoxides (LH), the ferric reducing ability of plasma (FRAP), superoxide dismutase (SOD), and catalase activity. CRP [1.8 ± 0.5 mg/L (MS), 1.0 ± 0.5 mg/L (C)] and LH [1.2 ± 0.2 μmol/L (MS), 1.1 ± 0.1 μmol/L (C)] were not different between MS patients and controls. FMD [8.0 ± 1.2% (MS) and 9.2 ± 1.6% (C)] and reactive hyperemia [380 ± 61 mL (MS) and 402 ± 69 mL (C)] were also not different between groups. Vascular function, as assessed by both FMD and reactive hyperemia, was not impaired in patients with MS compared to controls. Further, there was no evidence of elevated systemic inflammation or oxidative stress in these patients, who were currently all in remission. These findings suggest that impaired vascular function, elevated inflammation and oxidative stress are not an obligatory accompaniment to MS.


Inducible gene manipulations in serotonergic neurons.
An impairment of the serotonergic (5-HT) system has been implicated in the etiology of many neuropsychiatric disorders. Despite the considerable genetic evidence, the exact molecular and pathophysiological mechanisms underlying this dysfunction remain largely unknown. To address the lack of instruments for the molecular dissection of gene function in serotonergic neurons we have developed a new mouse transgenic tool that allows inducible Cre-mediated recombination of genes selectively in 5-HT neurons of all raphe nuclei. In this transgenic mouse line, the tamoxifen-inducible CreERT2 recombinase is expressed under the regulatory control of the mouse tryptophan hydroxylase 2 (Tph2) gene locus (177 kb). Tamoxifen treatment efficiently induced recombination selectively in serotonergic neurons with minimal background activity in vehicle-treated mice. These genetic manipulations can be initiated at any desired time during embryonic development, neonatal stage or adulthood. To illustrate the versatility of this new tool, we show that Brainbow-1.0L(TPH2-CreERT2) mice display highly efficient recombination in serotonergic neurons with individual 5-HT neurons labeling with multiple distinct fluorescent colors. This labeling is well suited for visualization and tracing of serotonergic neurons and their network architecture. Finally, the applicability of TPH2-CreERT2 for loxP-flanked candidate gene manipulation is evidenced by our successful knockout induction of the ubiquitously expressed glucocorticoid-receptor exclusively in 5-HT neurons of adult mice. The TPH2-CreERT2 line will allow detailed analysis of gene function in both developing and adult serotonergic neurons.


Low-Voltage Organic Transistor Memory Fiber with a Nanograined Organic Ferroelectric Film.
Wearable technology offers new ways to be more proactive about our health and surroundings in real time. For next-generation wearable systems, robust storage and recording media are required to monitor and process the essential electrical signals generated under various unpredictable strain conditions. Here, we report the first fibriform organic transistor memory integrated on a thin and flexible metal wire. A capillary tube coating system allows the formation of a thin and nanograined organic ferroelectric film on the wire. The uniform morphology imparts excellent switching stability (∼100 cycles), quasi-permanent retention (over 5 × 104 s), and low-voltage operation (below 5 V) to the fiber-shaped memory devices. When sewn in a stretchable textile fabric, the memory fiber achieves long retention time of more than 104 s with negligible degradation of memory window even under a constant diagonal strain of 100% that exhibits reliable data storage under tough environments. These results illustrate the possibility of the practical, wearable fiber memory for recording electronic signals in smart garment applications.


The Pseudomonas aeruginosa dnaK gene is involved in bacterial translocation across the intestinal epithelial cell barrier.
Pseudomonas aeruginosa can penetrate through polarized epithelial cell monolayers produced by the human adenocarcinoma cell line Caco-2. We previously identified genes associated with bacterial translocation through Caco-2 cell monolayers by analysing transposon insertion mutants with dramatically reduced penetration activity relative to that of the wild-type P. aeruginosa PAO1 strain. In this study, we focused on the dnaK mutant because the association between this gene and penetration activity is unknown. Inactivation of dnaK caused significant repression of bacterial penetration through Caco-2 cell monolayers, with decreased swimming, swarming and twitching motilities; bacterial adherence; and fly mortality rate; as well as dramatic repression of type III effector secretion and production of elastase and exotoxin A. However, type IV pilus protein PilA expression was not affected. These results suggest that dnaK is associated with bacterial motility and adherence, which are mediated by flagella and pili, and with toxin secretion, which plays a key role in the penetration of P. aeruginosa through Caco-2 cell monolayers. Inactivation of P. aeruginosa dnaK function may interfere with bacterial translocation and prevent septicaemia caused by P. aeruginosa.


Further characterization of the quinine taste discrimination stimulus in rhesus monkeys: effects of inter-trial interval, number and distribution of test stimuli.
The purpose of the current investigation was to characterize further the taste discriminative stimulus properties of quinine. Using a food-maintained two-choice (quinine and water) discrimination paradigm, the effects of varying inter-trial intervals, and the number and distribution of test stimuli were examined in rhesus monkeys (Macaca mulatta). Monkeys were trained to respond under a chain schedule of reinforcement (FR5B-fluid, FR30B-food) with a 60-s inter-trial interval. Interestingly, the quinine discrimination remained invariant across all the conditions tested: alterations in inter-trial interval (0, 15, 30, 45, 60 s), alterations in the number of test stimuli (six at quarter-log unit increments [0.1-1 mg/ml] versus 10 at quarter-log unit increments [0.01-1 mg/ml]), and alterations in the distribution of the test stimuli (quarter-log unit increments [0.1-0.1 mg/ml] versus half-log unit increments [0.01-1 mg/ml]) failed to alter the quinine discriminative stimulus (ED50s ranged from 0.18-0.29 mg/ml). Furthermore, incorrect lever selections were very rare (less than 12% of all test sessions), independent of test stimuli order (i.e. quinine followed by water trials, water followed by quinine trials), and independent of the progress through the test session (i.e. incorrect lever selections were consistent--approximately one error for every 10 trials--within early, middle and late portions of the session). These results demonstrate the reliability and robustness of the quinine taste discriminative stimulus in rhesus monkeys and provide further validation for the experimental paradigm.


Cajal body proteins SMN and Coilin show differential dynamic behaviour in vivo.
Analysis of stable cell lines expressing fluorescently tagged survival of motor neurons protein (SMN) and coilin shows striking differences in their dynamic behaviour, both in the nucleus and during mitosis. Cajal bodies labelled with either FP-SMN or FP-coilin show similar behaviour and frequency of movements. However, fluorescence recovery after photobleaching (FRAP) studies show that SMN returns approximately 50-fold more slowly to Cajal bodies than does coilin. Time-lapse studies on cells progressing from prophase through to G1 show further differences between SMN and coilin, both in their localisation in telophase and in the timing of their re-entry into daughter nuclei. The data reveal similarities between Cajal bodies and nucleoli in their behaviour during mitosis. This in vivo study indicates that SMN and coilin interact differentially with Cajal bodies and reveals parallels in the pathway for reassembly of nucleoli and Cajal bodies following mitosis.


Effect of locally produced complementary foods on fat-free mass, linear growth, and iron status among Kenyan infants: A randomized controlled trial.
The impact of quality complementary food products on infant growth and body composition has not been adequately investigated. This study evaluated the effect on fat-free mass (FFM) accrual, linear growth, and iron status of locally produced complementary food products comparing to a standard product. In a randomized, double-blind trial, 499 infants at 6 months received nine monthly rations of (a) WinFood Classic (WFC) comprising germinated amaranth (71%), maize (10.4%), small fish (3%), and edible termites (10%); (b) WinFood Lite (WFL) comprising germinated amaranth (82.5%), maize (10.2%), and multimicronutrient premix; or (c) fortified corn-soy blend plus (CSB+). Primary outcomes were changes in FFM, length, and plasma ferritin and transferrin receptors (TfR). FFM was determined using deuterium dilution. Analysis was by intention to treat, based on available cases. Compared with CSB+, there were no differences in change from 6 to 15 months in FFM for WFC 0.0 kg (95% CI [-0.30, 0.29]) and WFL 0.03 kg (95% CI [-0.25, 0.32]) and length change for WFC -0.3 cm (95% CI [-0.9, 0.4]) and WFL -0.3 cm (95% CI [-0.9, 0.3]). TfR increased in WFC group 3.3 mg L-1 (95% CI [1.7, 4.9]) and WFL group 1.7 mg L-1 (95% CI [0.1, 3.4]) compared with CSB+. Compared with the increase in Hb in CSB+ group, there was a reduction in Hb in WFC of -0.9 g dl-1 (95% CI [-1.3, -0.5]) and a lower increase in WFL -0.4 g dl-1 (95% CI [-0.8, 0.0]). In conclusion, the tested WinFoods had the same effect on FFM and length as CSB+, whereas Hb and iron status decreased, suggesting inhibited iron bioavailability from the amaranth-based WinFoods.


Spinal cord injury-induced lesional expression of the repulsive guidance molecule (RGM).
The repulsive guidance molecule (RGM) is involved in the formation of the central nervous system (CNS) during development by modulating guidance of growing axons. However, a role of RGM in CNS injury remains to be established. We studied the expression of RGM in the spinal cord of rats with spinal cord injury (SCI). After SCI, RGM+ cells accumulated in lesions and peri-lesional areas. During the first days after SCI, RGM expression was confined to neurons, ballooned neurite fibers/retraction bulbs, smooth muscle/endothelial cells, and to leucocytes infiltrating the lesion. Lesional RGM expression was frequently confined to hypertrophic beta-APP+ and RhoA+ neurites/retraction bulbs. With maturation of the lesion, we observed RGM expression by components of the developing scar tissue (cicatrix), such as fibroblastoid cells, reactive astrocytes and in addition a pronounced extracellular RGM deposition resembling neo-laminae. Frequent RGM+, RhoA+ coexpression by lesional retraction bulbs represent first preliminary evidence of RGM to exert growth inhibitory effects by the second messenger system RhoA. To date, RGM is one of the most potent axonal growth inhibitors identified and present in axonal growth impediments (i) oligodendrocytes; (ii) the plexus choroideus and (iii) components of the developing scar.


The role of microRNAs in solving COVID-19 puzzle from infection to therapeutics: A mini-review.
Nowadays, one of the major global health concerns is coronavirus disease 2019 (COVID-19), which is caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Even though numerous treatments and vaccines to combat this virus are currently under development, the detailed molecular mechanisms underlying the pathogenesis of this disease are yet to be elucidated to design future therapeutic tools against SARS-CoV-2 variants. MicroRNAs (miRNAs) are small (20-24 nucleotides), non-coding RNA molecules that regulate post-transcriptional gene expression. Recently, it has been demonstrated that both host and viral-encoded miRNAs are crucial for the successful infection of SARS-CoV-2. For instance, dysregulation of miRNAs that modulate multiple genes expressed in COVID-19 patients with comorbidities (e.g., type 2 diabetes, lung adenocarcinoma, and cerebrovascular disorders) could affect the severity of the disease. Thus, altered expression levels of circulating miRNAs might be helpful to diagnose this illness and forecast whether a COVID-19 patient could develop a severe state of the disease. Besides, researchers have found a number of miRNAs could inhibit the expression of proteins, such as ACE2, TMPRSS2, spike, and Nsp12, involved in the life cycle of SARS-CoV-2. Accordingly, miRNAs represent potential biomarkers and therapeutic targets for this devastating viral disease. Therefore, in this current review, we present the recent discoveries regarding the clinical relevance and biological roles of miRNAs in COVID-19.


A highly effective and stable bispecific diabody for cancer immunotherapy: cure of xenografted tumors by bispecific diabody and T-LAK cells.
In the field of cancer immunotherapy research, the targeting of effector cells with specific antibodies is a very promising approach. Recent advances in genetic engineering have made it possible to prepare immunoglobulin fragments consisting of variable domains using bacterial expression systems. We have produced an anti-epidermal growth-factor receptor (EGFR) x anti-CD3 bispecific diabody (Ex3 diabody) in an Escherichia coli (E. coli) expression system with refolding method. The Ex3 diabody targets lymphokine-activated killer cells with a T-cell phenotype (T-LAK cells) to EGFR positive bile duct carcinoma cells with dramatic enhancement of cytotoxicity in vitro. This specific killing of EGFR-positive cells was completely inhibited by parental mAb IgGs directed to EGFR and the CD3 antigen. When T-LAK cells were cultured with EGFR-positive tumor cells in the presence of Ex3 diabody, they produced much higher levels of IFN-gamma, GM-CSF, and TNF-alpha than in its absence, this being a possible mechanism underlying specific antitumor activity. The Ex3 diabody showed good stability when tested at 37 degrees C for 48 h, and also markedly inhibited tumor growth of bile duct carcinoma xenografts in severe combined immunodeficient (SCID) mice. When Ex3 diabody (20 microg/mouse) was administrated intravenously, together with T-LAK cells and interleukin-2 (IL-2), complete cure of tumors were observed in three of six mice, and the other three showed marked retardation of tumor growth. The Ex3 diabody can be considered a highly promising reagent for study of specific targeting immunotherapy against bile duct and other EGFR-positive carcinomas.


Postoperative thrombocytopenia and subsequent consequences in acute type A aortic dissection.
To ascertain if postoperative thrombocytopenia following open aortic surgery with a median sternotomy can predict early- and intermediate-term morbidity and mortality. From January 2018 to December 2022, a comparison was made between patients who had and didn't have postoperative thrombocytopenia (defined as a nadir < 75 × 103/μL after 72 h of open aortic surgery with median sternotomy). Intermediate-term mortality during follow-up was the main result, with cerebrovascular accident and acute renal injury requiring dialysis as secondary events. Inverse probability treatment weighting (IPTW) was used to account for selection bias between groups. The Kaplan-Meier method with the log-rank test was used to assess intermediate-term survivals following IPTW modification. To identify the nonlinear link between platelet nadir and mortality probability, a generalized additive mix model was applied. To help increase power in testing for the overall effect of platelet nadir on outcomes in the generalized additive mix model, the hazard ratios and 95% CIs for each subgroup and their interactions were examined. The study included 457 patients, 347 male (75.9%), with mean age of 54 ± 12 years. The last follow-up was done on April 14th, 2023 and the median follow-up time was 16 (6-31) months. Following IPTW, patient characteristics were balanced among cohorts. Platelet nadir was found to be significantly inversely related to early-term mortality (IPTW-adjusted hazard ratio = 0.968 (0.960, 0.977), p < 0.001), and AKI requiring dialysis (IPTW-adjusted hazard ratio = 0.979 (0.971, 0.986), p < 0.001). A nonlinear relationship between platelet nadir and mortality risk probability during follow-up visually showed that the likelihood of mortality decreased with platelet nadir increased. In confounder-adjusted survival ('postoperative thrombocytopenia not acquired' vs 'postoperative thrombocytopenia'; HR: 0.086 [95% CI: 0.045-0.163]; p < 0.01) analysis, non-acquired postoperative thrombocytopenia was associated with a lower risk of mortality, and the treatment benefit was validated in IPTW-adjusted analysis, which showed an HR of 0.067. Early postoperative thrombocytopenia following type A aortic dissection surgery is a risk factor for morbidity and mortality. Because postoperative thrombocytopenia can indicate a poor prognosis, monitoring early postoperative platelets helps identify individuals who may develop late postoperative problems, which is performed by this affordable biomarker.


Assessing public awareness of clubfoot and knowledge about the importance of early childhood treatment: a cross-sectional survey.
Clubfoot is a treatable abnormality that can be managed with early intervention. However, there is a lack of public knowledge regarding clubfoot, which can delay treatment. This study aimed to assess the public awareness of clubfoot and knowledge regarding the importance of treatment in early childhood. This cross-sectional survey spanned 6 months, from June through November 2018, and involved persons living in Saudi Arabia. To collect data on public awareness of clubfoot risk factors, treatment, and prognosis, a questionnaire was developed by orthopedic experts and disseminated online. The target population included people of both genders and all age groups from the general population, regardless of their knowledge of someone with clubfoot. By the end of the study period, 746 participants completed the online survey. In total, 520 of the respondents (69.7%) had never heard about clubfoot syndrome. Among the participants, 5.4% had a child with clubfoot syndrome and 4.6% were aware of clubfoot because they had an affected child. The top resource accessed by respondents for obtaining knowledge about clubfoot was social media channels (38.4%), followed by obtaining knowledge from relatives and friends (19.9%). The most reported perceived cause of clubfoot was hereditary and genetic disorders (58.4%), followed by neurological disorders (39.9%). Results show that there is low public knowledge of clubfoot which may be attributed to a lack of awareness campaigns. We recommend increasing awareness regarding clubfoot through social media platforms and public campaigns in key locations, such as malls, as this may encourage people to seek early treatment. This is important because early management of clubfoot is less invasive and with regular follow-up, leads to better patient outcomes.


Patterns of genetic variation in populations of infectious agents.
The analysis of genetic variation in populations of infectious agents may help us understand their epidemiology and evolution. Here we study a model for assessing the levels and patterns of genetic diversity in populations of infectious agents. The population is structured into many small subpopulations, which correspond to their hosts, that are connected according to a specific type of contact network. We considered different types of networks, including fully connected networks and scale free networks, which have been considered as a model that captures some properties of real contact networks. Infectious agents transmit between hosts, through migration, where they grow and mutate until elimination by the host immune system. We show how our model is closely related to the classical SIS model in epidemiology and find that: depending on the relation between the rate at which infectious agents are eliminated by the immune system and the within host effective population size, genetic diversity increases with R0 or peaks at intermediate R0 levels; patterns of genetic diversity in this model are in general similar to those expected under the standard neutral model, but in a scale free network and for low values of R0 a distortion in the neutral mutation frequency spectrum can be observed; highly connected hosts (hubs in the network) show patterns of diversity different from poorly connected individuals, namely higher levels of genetic variation, lower levels of genetic differentiation and larger values of Tajima's D. We have found that levels of genetic variability in the population of infectious agents can be predicted by simple analytical approximations, and exhibit two distinct scenarios which are met according to the relation between the rate of drift and the rate at which infectious agents are eliminated. In one scenario the diversity is an increasing function of the level of transmission and in a second scenario it is peaked around intermediate levels of transmission. This is independent of the type of host contact structure. Furthermore for low values of R0, very heterogeneous host contact structures lead to lower levels of diversity.


Immunostimulatory CpG-DNA activates murine microglia.
Bacterial DNA containing motifs of unmethylated CpG dinucleotides (CpG-DNA) triggers innate immune cells through the pattern recognition receptor Toll-like receptor 9 (TLR-9). CpG-DNA possesses potent immunostimulatory effects on macrophages, dendritic cells, and B lymphocytes. Therefore, CpG-DNA contributes to inflammation during the course of bacterial infections. In contrast to other TLR-dependent microbial patterns, CpG-DNA is a strong inductor of IL-12. Thus, it acts as a Th1-polarizing agent that can be utilized as potent vaccine adjuvant. To assess the role of CpG-DNA in immune reactions in the CNS, we analyzed the effects of CpG-DNA on microglial cells in vitro and in vivo. Primary microglial cells as well as microglial cell lines express TLR-9 mRNA. Consequently, CpG-DNA activated microglial cells in vitro and induced TNF-alpha, IL-12p40, IL-12p70, and NO. Furthermore, MHC class II, B7-1, B7-2, and CD40 molecules were up-regulated. In addition, phagocytic activity of microglia was enhanced. After intracerebroventricular injection of CpG-DNA, microglial cells were activated and produced TNF-alpha and IL-12p40 transcripts, as shown by in situ hybridization. These results indicate that microglia is sensitive to CpG-DNA. Thus, bacterial DNA containing CpG motifs could not only play an important role during infections of the CNS, but also might trigger and sustain Th1-dominated immunopathogenic reactions.


Acute uvulitis in combination with acute epiglottitis: a case presentation.
Acute uvulitis in adults is a rare condition, especially when associated with acute epiglottitis. Watchful monitoring of the airway, usually in the intensive care unit, is recommended. Along with appropriate antibiotics and steroids, we recommend all patients with acute uvulitis have a lateral radiograph of the neck to rule out acute epiglottitis. To avoid a possible fatality, a high index of suspicion should be maintained at all times in order to appropriately diagnose and treat these patients.


Ruthenium-catalyzed α-(hetero)arylation of saturated cyclic amines: reaction scope and mechanism.
Transition-metal-catalyzed sp(3) C-H activation has emerged as a powerful approach to functionalize saturated cyclic amines. Our group recently disclosed a direct catalytic arylation reaction of piperidines at the α position to the nitrogen atom. 1-(Pyridin-2-yl)piperidine could be smoothly α-arylated if treated with an arylboronic ester in the presence of a catalytic amount of [Ru3(CO)12] and one equivalent of 3-ethyl-3-pentanol. A systematic study on the substrate and reagent scope of this transformation is disclosed in this paper. The effect of substitution on both the piperidine ring and the arylboronic ester has been investigated. Smaller (pyrrolidine) and larger (azepane) saturated ring systems, as well as benzoannulated derivatives, were found to be compatible substrates with the α-arylation protocol. The successful use of a variety of heteroarylboronic esters as coupling partners further proved the power of this direct functionalization method. Mechanistic studies have allowed for a better understanding of the catalytic cycle of this remarkable transformation featuring an unprecedented direct transmetalation on a Ru(II)-H species.


Risk of therapy-related myelodysplastic syndrome/acute leukemia following high-dose therapy and autologous bone marrow transplantation for non-Hodgkin's lymphoma.
Several recent reports have suggested that patients with non-Hodgkin's lymphomas (NHL) who undergo autologous stem cell transplantation (ASCT) are at increased risk of developing therapy-related myelodysplastic syndrome (tMDS) and acute myelogenous leukemia (tAML). We analyzed 493 patients with NHL who underwent ASCT at The University of Texas M.D. Anderson Cancer Center between January 1990 and August 1999. With a median follow-up time of 21 months after HDT, 22 patients developed persistent cytopenia in at least one cell line with morphologic or cytogenetic evidence of tMDS or tAML. Univariate analysis identified prior fludarabine therapy, bone marrow involvement with lymphoma, and total body irradiation (TBI) as significant risk factors for the development of tMDS/tAML (P <0.05). Multiple logistic regression analysis showed that TBI was independently associated with an increased risk of developing tMDS/tAML (P <0.01). Further analysis of the patients who received TBI revealed that patients receiving TBI in combination with cyclophosphamide and etoposide were more likely to develop tMDS/tAML than those who received TBI with cyclophosphamide or thiotepa (P <0.01). The median survival of patients developing tMDS/tAML was 7.5 months (range 0-32 months). TBI, especially when used in combination with cyclophosphamide and etoposide as the pretransplant conditioning regimen, is a significant risk factor for the development of tMDS/tAML.


Inter- and intra-observer agreement of femoroacetabular impingement (FAI) parameters comparing plain radiographs and advanced, 3D computed tomographic (CT)-generated hip models in a surgical patient cohort.
The purpose of our study was to investigate whether advanced, 3D computed tomographic (CT)-generated hip models improves inter-and intra-observer agreement when compared to plain radiographs in identifying femoroacetabular impingement (FAI) morphology. Eight consecutive patients who underwent surgery for FAI pathology were selected for this study. Preoperative CT scan image data were used to create high resolution, 3D hip reconstruction models. Four observers (two attending hip surgeons and radiologists) performed a blinded review of preselected radiographs and 3D CT hip models. Alpha and lateral center-edge angle measurements, location of cam lesion and the presence of a "crossover sign" were assessed. Inter- and intra-observer agreement was determined by calculating the intra-class correlation coefficients (ICC) or kappa coefficients to evaluate agreement for categorical variables. The parameter that demonstrated the highest and poorest inter-observer agreement was the presence of a "crossover sign" using 3D CT-generated high resolution hip models (ICC = 0.76, p = 0.00) and anteroposterior pelvis radiography, respectively (ICC = 0.20, p = 0.02). Alpha angle values were significantly higher using plain radiographs when compared to 3D hip reconstruction models (61.1° ± 10.4° versus 55.4° ± 14.4°, p = 0.003). Furthermore, when compared to radiographs, 3D hip reconstruction models demonstrated significantly higher intra-observer agreement (ICC = 0.856 versus 0.405, p = 0.005) when determining the presence of a "crossover sign". Our findings were suggestive that for most commonly used FAI morphology parameters, CT-generated hip models demonstrated little benefit over plain radiographs in improving inter-observer agreement among providers. III.


Mechanism-Driven Metabolic Engineering for Bio-Based Production of Free R-Lipoic Acid in Saccharomyces cerevisiae Mitochondria.
Lipoic acid is a valuable organosulfur compound used as an antioxidant for dietary supplementation, and potentially anti-diabetic and anti-cancer. Currently, lipoic acid is obtained mainly through chemical synthesis, which requires toxic reagents and organic solvents, thus causing environmental issues. Moreover, chemically synthesized lipoic acid is conventionally a racemic mixture. To obtain enantiomerically pure R-lipoic acid, which has superior bioactivity than the S form, chiral resolution and asymmetric synthesis methods require additional reagents and solvents, and often lead to wastage of S-lipoic acid or precursors with undesired chirality. Toward sustainable production of R-lipoic acid, we aim to develop a synthetic biology-based method using engineered yeast. Here, we deepened mechanistic understanding of lipoic acid biosynthesis and protein lipoylation in the model yeast Saccharomyces cerevisiae to facilitate metabolic engineering of the microbe for producing free R-lipoic acid. In brief, we studied the biosynthesis and confirmed the availability of protein-bound lipoate in yeast cells through LC-MS/MS. We then characterized in vitro the activity of a lipoamidase from Enterococcus faecalis for releasing free R-lipoic acid from lipoate-modified yeast proteins. Overexpression of the lipoamidase in yeast mitochondria enabled de novo free R-lipoic acid production in vivo. By overexpressing pathway enzymes and regenerating the cofactor, the production titer was increased ∼2.9-fold. This study represents the first report of free R-lipoic acid biosynthesis in S. cerevisiae. We envision that these results could provide insights into lipoic acid biosynthesis in eukaryotic cells and drive development of sustainable R-lipoic acid production.


Amino acid uptake and regulation in multicellular hepatoma spheroids.
Cancer cells maintained in monolayer tissue culture are frequently used to study tumor biology and nutrient uptake, but there is a concern that this system may not fully reflect clinical tumor physiology. Because cells grown in a 3-D configuration more closely resemble an in vivo environment, a model was developed and characterized for the growth of SK-Hep human hepatoma cells in suspension as multicellular tumor spheroids (MTS). The measurement of nutrient uptake in such a system has never been established. SK-Hep cultures were initiated as single cell suspensions and grown as MTS in siliconized spinner flasks. The transport of several individual amino acids (arginine, glutamate, leucine, alpha-(N-methylamino)isobutyric acid (MeAIB), and glutamine (GLN)) was measured in SK-Hep single cell suspensions and MTS (0. 50-0.60 mm diameter) by a radiotracer/rapid filtration technique, as was the regulation of glutamine uptake by phorbol esters. l-[(3)H]GLN uptake was also measured in larger spheroids (0.85-1.5 mm diameter). MTS cellularity was evaluated by histological examination, and single cell integrity after the transport assay was confirmed by scanning electron microscopy (SEM). SK-Hep MTS displayed gradients of cellular morphology and staining, with central necrosis visible at diameters >0.8 mm. Single cell suspensions endured the rapid filtration technique based on functional Na(+)-dependent uptake rates and SEM analysis. Of all amino acids tested, only GLN transport rates were visibly affected by growth format. In small MTS, Na(+)-dependent GLN uptake was diminished by 40%, but was 40-53% higher in MTS >1 mm displaying central necrosis, when compared to single cell suspensions. Likewise, slight parallel changes in glutamine transporter ATB(0) mRNA levels were observed in Northern blot analysis. Finally, phorbol ester-dependent GLN transport down-regulation (by 40-50%), previously established in SK-Hep monolayers, remained operative in all cell formats tested. The data suggest that the tumor microenvironment differentially impacts the uptake of specific nutrients despite the conservation of key regulatory pathways. This MTS technique may prove useful for further studies on the role of nutrient transport in nascent tumor growth.


Exergy-based efficiency and renewability assessment of biofuel production.
This study presents an efficiency and renewability analysis of the production of three biofuels: rapeseed methyl ester (RME), soybean methyl ester (SME) and corn-based ethanol (EtOH). The overall production chains have been taken into account: not only the agricultural crop production and the industrial conversion into biofuel, but also production of the supply of agricultural resources (pesticides, fertilizers, fuel, seeding material) and industrial resources (energy and chemicals) to transform the crops into biofuel. Simultaneously, byproducts of the agricultural and industrial processes have been taken into account when resources have to be allocated to the biofuels. The technical analysis via the second law of thermodynamics revealed that corn-based EtOH results in the highest production rate with an exergetic fuel content of 68.8 GJ ha(-1) yr(-1), whereas the RME and SME results were limited to 47.5 and 16.4 GJ ha(-1) yr(-1). The allocated nonrenewable resource input to deliver these biofuels is significant: 16.5, 15.4, and 5.6 MJ ha(-1) yr(-1). This means that these biofuels, generally considered as renewable resources, embed a nonrenewable fraction of one-quarter for EtOH and even one-third for RME and SME. This type of analysis provides scientifically sound quantitative information that is necessarywith respect to the sustainability analysis of so-called renewable energy.


Isometric knee extension force measured using a handheld dynamometer with and without belt-stabilization.
Although evidence suggests that tester strength limits the magnitude of isometric force that can be measured using a handheld dynamometer (HHD), previous studies have not investigated the actual limits of force magnitude that can be measured by trained testers when a belt is or is not used to stabilize the dynamometer. Therefore, the primary aims of this study were to determine: 1) the magnitude of knee extension forces that could be measured with a HHD with and without belt-stabilization and 2) the relationship between tester characteristics and knee extension strength measured with and without belt-stabilization. The characteristics of 20 trained testers (10 men, 10 women) were determined. Thereafter, they measured isometric knee extension strength using the MicroFET HHD with and without belt-stabilization. Paired t-tests were used to compare maximal knee extension forces under two conditions. Pearson product-moment correlations were calculated to determine the relationship between tester characteristics and knee extension forces measured under the two conditions. Knee extension forces (Newtons) measured using the HHD without belt-stabilization (470.6 ± 179.8) were significantly lower (t= -7.968, p<0.001) than those measured with belt-stabilization (866.9 ± 131.7). Pearson correlations between tester characteristics and knee extension forces measured with no belt-stabilization were all statistically significant (p ≤ 0.002); however, the correlations were not statistically significant under the belt-stabilization condition. The forces that can be measured with a HHD are higher than those suggested by previous researchers. By rectifying limitations imposed by tester strength, use of a belt allows very high knee extension forces to be measured.


Identification of Burkholderia species and genomovars from cystic fibrosis patients by AFLP fingerprinting.
AFLP is a genomic fingerprinting technique based on the selective amplification of restriction fragments from a total double-digest of genomic DNA. The applicability of this method to differentiate between species and genomovars of the genus Burkholderia was tested, with particular emphasis on taxa occurring in cystic fibrosis patients. In this study, 78 well-characterized strains and field isolates were investigated by two methods of AFLP fingerprinting. In the manual procedure, a radioactively labelled primer was used, amplified fragments were separated by conventional PAGE and the patterns were revealed by autoradiography. In the automated procedure, a fluorescently labelled primer was used in combination with electrophoresis and on-line data collection by means of an automated DNA sequencer. Overall, there was good agreement between the two AFLP procedures and the data were mostly consistent with results obtained from SDS-PAGE of whole-cell proteins and DNA-DNA hybridization experiments. The automated AFLP procedure has considerable technical advantages compared with the manual AFLP procedure, but a thorough visual analysis of the DNA profiles was required to avoid misidentification of some Burkholderia cepacia genomovar III strains.


Why do Danish adolescents take up smoking?
For the first time a comprehensive international research-based smoking prevention programme addressing adolescents has been launched in Denmark. The ESFA cohort study started in 1997, and this article presents baseline findings. Factors that relate to the initiation of adolescent smoking are found among the concepts of attitude, social norms, social pressure and self-efficacy beliefs. Baseline findings from Danish adolescents (n = 1770) in secondary school (mean age 13.8 years). The cross-sectional baseline data show that the attitude-social influence-efficacy-model proves to be of value in highlighting the associations of adolescent smoking behaviour. Social self-efficacy, peer smoking behaviour and the intention to smoke proved to have the strongest associations with smoking. It is recommended that self-efficacy beliefs and the individual intentions to smoke should be included in the future in smoking prevention programmes in Denmark. Furthermore, a distinction between the roles of parents and peers should be emphasized when addressing the social environment of adolescents in health promotion.


Pravastatin for preeclampsia: From animal to human.
Preeclampsia is characterized by the emergence of hypertension and proteinuria after 20 weeks of pregnancy, and it threatens both maternal and fetal lives if it proceeds unabated. Despite numerous studies, thus far the only fundamental therapy for preeclampsia is termination of pregnancy, leading to preterm birth. Furthermore, preeclamptic women are reported to be at risk for cardiovascular diseases for 10 years after delivery. Therefore, preventative and therapeutic strategies for preeclampsia are required. Recently, statins have been reported to improve the regeneration of vascular endothelium, and pravastatin has attracted attention as a potential preventative or therapeutic candidate for preeclampsia. Pravastatin has been demonstrated to have preventative effects in preeclampsia model mice, and a large volume of human data from pregnant women taking statins supports the safety of these drugs. Moreover, small clinical trials have reported that pravastatin has strong preventative or therapeutic effects on preeclampsia and it has the potential to improve the prognosis of pregnant women, fetuses and neonates affected by this condition.


White matter hyperintensities mediate the association of nocturnal blood pressure with cognition.
To test the hypotheses that hypertension and nocturnal blood pressure are related to white matter hyperintensity (WMH) volume, an MRI marker of small vessel cerebrovascular disease, and that WMH burden statistically mediates the association of hypertension and dipping status with memory functioning, we examined the relationship of hypertension and dipping status on WMH volume and neuropsychological test scores in middle-aged and older adults. Participants from the community-based Maracaibo Aging Study received ambulatory 24-hour blood pressure monitoring, structural MRI, and neuropsychological assessment. Four hundred thirty-five participants (mean ± SD age 59 ± 13 years, 71% women) with available ambulatory blood pressure, MRI, and neuropsychological data were included in the analyses. Ambulatory blood pressure was used to define hypertension and dipping status (i.e., dipper, nondipper, and reverse dipper based on night/day blood pressure ratio <0.9, 0.9-1, and >1, respectively). Outcome measures included regional WMH and memory functioning derived from a neuropsychological test battery. The majority of the participants (59%) were hypertensive. Ten percent were reverse dippers, and 40% were nondippers. Reverse dipping in the presence of hypertension was associated with particularly elevated periventricular WMH volume (F 2,423 = 3.78, p = 0.024) and with lowered memory scores (F 2,423 = 3.911, p = 0.021). Periventricular WMH volume mediated the effect of dipping status and hypertension on memory (β = -4.1, 95% confidence interval -8.7 to -0.2, p < 0.05). Reverse dipping in the presence of hypertension is associated with small vessel cerebrovascular disease, which, in turn, mediates memory functioning. These results point toward reverse dipping as a marker of poor nocturnal blood pressure control, particularly among hypertensive individuals, with potentially pernicious effects on cerebrovascular health and associated cognitive function.


[Mechanism and implication of regulation of the pricing of essential medicines in the private pharmaceutical sector in Mali].
The healthcare and pharmaceutical professions in Mali were privatized in 1985. Privatization led to swift expansion of the private sector and upset the balance that had existed between the public and private sectors. A national pharmaceutical policy did not emerge until a decade later. Its purpose was to promote a system ensuring fair access to essential generic medicines for all. It was hoped that synergy between the two sectors would promote that objective. However, the policy calling for distribution of essential generic medicine through the private sector was not accompanied by an adequate system for pricing. This problem led the government to adopt a price regulation policy to realign market dynamics with public health goals. This experience shows that a sustained effort from public policy makers is necessary to prevail against the professional and business interests that can conflict with the public interest. Analysis of this experience also demonstrates the need to improve, restructure, and control the pharmaceutical industry. The government must continue to play its crucial role in the context of limited resources and inequality between consumers and pharmaceutical companies.


Growth hormone treatment in Aarskog syndrome: analysis of the KIGS (Pharmacia International Growth Database) data.
Aarskog syndrome is an X-linked disorder characterized by faciogenital dysplasia and short stature. The present study set out to determine the effect of growth hormone (GH) therapy in patients with Aarskog syndrome enrolled in KIGS--the Pharmacia International Growth Database. Twenty-one patients (20 males) were evaluated. Median age at start of treatment was 8.3 years (10-90th percentiles, 5.1-14.1 years) and median height SDS was -2.8 (10-90th percentiles, -2.1 to -3.7). The median dose of GH was 0.22 mg/kg/week (10-90th percentiles, 0.15-0.30 mg/kg/week) given at a median frequency of six (4-7) times per week. Prepubertal patients were followed longitudinally for 1 year (n = 13) or 3 years (n = 7). After 1 year, the median height SDS had improved from -2.8 to -2.3 in 13 patients. After 3 years, height SDS had improved significantly (p <0.05) to -1.8 (10-90th percentiles, -2.1 to -1.1) in the seven patients. No adverse events were noted. Although final height data for these patients are still awaited, the present results support the use of GH to promote growth in children with Aarskog syndrome.


Self-starvation through the ages: reflections on the pre-history of anorexia nervosa.
Recent publications have indicated that voluntary self-starvation is not a recently developed syndrome and that it has been reported throughout history. These prior forms of inedia are summarized and related to their historical and cultural contexts. On the basis of these data, some hypotheses are proposed regarding social influences on the vulnerability to eating disorders. To document and describe forms of eating disorders occurring prior to the formal medical description of anorexia nervosa in the late 19th century. Review of historical references to self-starvation, of recent publications on the history of eating disorders, and of articles describing cases of eating disorders occurring in the past. Forms of eating disorders have existed since ancient times varying in frequency, manifestations, and possible motivation. Certain sociocultural factors appear to foster or inhibit the frequency and type of eating disorders.


Multisensory guidance of orienting behavior.
We use both vision and audition when localizing objects and events in our environment. However, these sensory systems receive spatial information in different coordinate systems: sounds are localized using inter-aural and spectral cues, yielding a head-centered representation of space, whereas the visual system uses an eye-centered representation of space, based on the site of activation on the retina. In addition, the visual system employs a place-coded, retinotopic map of space, whereas the auditory system's representational format is characterized by broad spatial tuning and a lack of topographical organization. A common view is that the brain needs to reconcile these differences in order to control behavior, such as orienting gaze to the location of a sound source. To accomplish this, it seems that either auditory spatial information must be transformed from a head-centered rate code to an eye-centered map to match the frame of reference used by the visual system, or vice versa. Here, we review a number of studies that have focused on the neural basis underlying such transformations in the primate auditory system. Although, these studies have found some evidence for such transformations, many differences in the way the auditory and visual system encode space exist throughout the auditory pathway. We will review these differences at the neural level, and will discuss them in relation to differences in the way auditory and visual information is used in guiding orienting movements.


Comparative evaluation of the indigenous microbial diversity vs. drilling fluid contaminants in the NEEM Greenland ice core.
Demonstrating that the detected microbial diversity in nonaseptically drilled deep ice cores is truly indigenous is challenging because of potential contamination with exogenous microbial cells. The NEEM Greenland ice core project provided a first-time opportunity to determine the origin and extent of contamination throughout drilling. We performed multiple parallel cultivation and culture-independent analyses of five decontaminated ice core samples from different depths (100-2051 m), the drilling fluid and its components Estisol and Coasol, and the drilling chips collected during drilling. We created a collection of diverse bacterial and fungal isolates (84 from the drilling fluid and its components, 45 from decontaminated ice, and 66 from drilling chips). Their categorization as contaminants or intrinsic glacial ice microorganisms was based on several criteria, including phylogenetic analyses, genomic fingerprinting, phenotypic characteristics, and presence in drilling fluid, chips, and/or ice. Firmicutes and fungi comprised the dominant group of contaminants among isolates and cloned rRNA genes. Conversely, most Proteobacteria and Actinobacteria originating from the ice were identified as intrinsic. This study provides a database of potential contaminants useful for future studies of NEEM cores and can contribute toward developing standardized protocols for contamination detection and ensuring the authenticity of the microbial diversity in deep glacial ice.


Aging and the skin immune system.
To give a brief summary of age-related alterations that occur in the immune system (immunosenescence), with special regard to the skin immune system. MEDLINE and institutional libraries were searched for relevant articles on immunosenescence and corresponding key words. The immune system of aged animals and humans undergoes alterations that may account for an increased susceptibility to certain infections, autoimmune diseases, or malignancies. However, some data on the parameters of immunosenescence are controversial. This appears to be caused by variations in study designs or by the many external influences that superimpose on intrinsic alterations of the immune system. Well characterized are age-related changes of T cells and cell-mediated immunity. With advancing age, human and murine T cells undergo a shift from the naive to the memory phenotype, associated with a change in cytokine profile. The cells also reveal reductions in the proliferative response to activation, in diversity of the T-cell receptor antigen repertoire, and in cytolytic activity. B cells of aging individuals show a restricted diversity of their antibody repertoire due to a decline in somatic mutations, resulting in a reduced response to certain viral infections or vaccinations. The number of Langerhans cells appears to decline with age, contributing to a reduced rate of sensitizations. Macrophages and keratinocytes also undergo age-related changes, although these are less well characterized. They entail alterations in cytokine production, which could play a role in increased susceptibility to endotoxins in elderly individuals. With aging, the skin immune system shows a decline in its adaptive capability, one of its outstanding qualities. Manipulations to revert age-related dysfunctions are being tested and may help to extend a healthy life.


The impact of child symptom severity on depressed mood among parents of children with ASD: the mediating role of stress proliferation.
Stress proliferation (the tendency of stressors to engender additional stressors in other life domains) is explored in a sample of 68 parents of children identified with ASD. Regression analyses showed that parent depression was predicted by both child symptom severity and by stress proliferation and that stress proliferation partially mediated the effect of child symptom severity on parent depression. In addition, informal social support was found to reduce levels of parent stress proliferation and parent depression; however, contrary to the stress buffering hypothesis, the ameliorative effect of support on stress proliferation was shown to be greatest when reported child symptomatology was less (rather than more) severe. Study implications for future research and practice are discussed.


The predictive value of RS time for short term mortality in patients with acute pulmonary embolism.
Many studies have examined the capability of electrocardiography (ECG) changes to predict the severity and prognosis of patients with acute pulmonary embolism (APE). RS time in ECG is potentially valuable in evaluating the prognosis of APE. In our study, we aimed to assess the predictive value of RS time, which is a novel electrocardiographic parameter of one-month mortality of APE. This retrospective study included 216 patients who were diagnosed with APE by pulmonary computed tomography angiography. RS time was measured from the ECG (inferolateral leads) at the time of hospital admission using a computer program (imagej.nih.gov/ij/). The patients were divided into two groups according to the median values of RS time: the group with RS time ≤ 60 msec (n:108) and the group with RS time > 60 msec (n:108). The groups were compared in terms of mortality. In our study, the one-month mortality was 15.3% (33) in the patients hospitalized with APE. In the multivariate analysis, RS time prolongation (HR: 1.037; 95%CI: 1.005-1.065; p = .02) was independently correlated with mortality. The ROC curve analysis revealed that RS time > 64.8 msec predicted the one-month mortality in APE with a sensitivity of 68.6% and a specificity of 73.9% (AUC: 0.708; 95% CI: 0.643-0.768; p < .001). As a novel ECG parameter, RS time could be measured for each patient with APE. Prolongation of RS time could be a useful index for predicting the one-month mortality of patients diagnosed with APE.


HIV-1 Nef targets MHC-I and CD4 for degradation via a final common beta-COP-dependent pathway in T cells.
To facilitate viral infection and spread, HIV-1 Nef disrupts the surface expression of the viral receptor (CD4) and molecules capable of presenting HIV antigens to the immune system (MHC-I). To accomplish this, Nef binds to the cytoplasmic tails of both molecules and then, by mechanisms that are not well understood, disrupts the trafficking of each molecule in different ways. Specifically, Nef promotes CD4 internalization after it has been transported to the cell surface, whereas Nef uses the clathrin adaptor, AP-1, to disrupt normal transport of MHC-I from the TGN to the cell surface. Despite these differences in initial intracellular trafficking, we demonstrate that MHC-I and CD4 are ultimately found in the same Rab7(+) vesicles and are both targeted for degradation via the activity of the Nef-interacting protein, beta-COP. Moreover, we demonstrate that Nef contains two separable beta-COP binding sites. One site, an arginine (RXR) motif in the N-terminal alpha helical domain of Nef, is necessary for maximal MHC-I degradation. The second site, composed of a di-acidic motif located in the C-terminal loop domain of Nef, is needed for efficient CD4 degradation. The requirement for redundant motifs with distinct roles supports a model in which Nef exists in multiple conformational states that allow access to different motifs, depending upon which cellular target is bound by Nef.


Anti-Fn14-Conjugated Prussian Blue Nanoparticles as a Targeted Photothermal Therapy Agent for Glioblastoma.
Prussian blue nanoparticles (PBNPs) are effective photothermal therapy (PTT) agents: they absorb near-infrared radiation and reemit it as heat via phonon-phonon relaxations that, in the presence of tumors, can induce thermal and immunogenic cell death. However, in the context of central nervous system (CNS) tumors, the off-target effects of PTT have the potential to result in injury to healthy CNS tissue. Motivated by this need for targeted PTT agents for CNS tumors, we present a PBNP formulation that targets fibroblast growth factor-inducible 14 (Fn14)-expressing glioblastoma cell lines. We conjugated an antibody targeting Fn14, a receptor abundantly expressed on many glioblastomas but near absent on healthy CNS tissue, to PBNPs (aFn14-PBNPs). We measured the attachment efficiency of aFn14 onto PBNPs, the size and stability of aFn14-PBNPs, and the ability of aFn14-PBNPs to induce thermal and immunogenic cell death and target and treat glioblastoma tumor cells in vitro. aFn14 remained stably conjugated to the PBNPs for at least 21 days. Further, PTT with aFn14-PBNPs induced thermal and immunogenic cell death in glioblastoma tumor cells. However, in a targeted treatment assay, PTT was only effective in killing glioblastoma tumor cells when using aFn14-PBNPs, not when using PBNPs alone. Our methodology is novel in its targeting moiety, tumor application, and combination with PTT. To the best of our knowledge, PBNPs have not been investigated as a targeted PTT agent in glioblastoma via conjugation to aFn14. Our results demonstrate a novel and effective method for delivering targeted PTT to aFn14-expressing tumor cells via aFn14 conjugation to PBNPs.


Differential Retinoic Acid Signaling in the Hippocampus of Aged Rats with and without Memory Impairment.
Retinoic acid (RA), a metabolite of vitamin A, has many physiological functions, and mounting evidence points to important roles in cognition. In vitro experiments indicate that RA is involved in homeostatic synaptic scaling in the hippocampus, which supports overall network stability during learning. It has been previously determined that disrupted RA signaling in the hippocampus causes deterioration of memory, that RA signaling declines with age in brain, and that application of RA reverses this decline. Here, we explore whether RA signaling is altered in an animal model of neurocognitive aging. We used a Morris water maze protocol to study cognitive decline in aged rats, which assesses hippocampus-dependent spatial memory and reveals substantial interindividual differences in aged animals. Aged unimpaired (AU) rats perform on par with young (Y), while aged impaired (AI) animals exhibit spatial memory deficits. We show that the major substrate for RA, retinol binding protein 4 (RBP4), is decreased in AU rats, and retinol cell surface receptor declines with chronological age. Other affected components of RA signaling include selective increases in AI animals in hippocampal synthesis (RALDH1) and catabolism of RA (CYP26B1), RA receptor α, the RA regulated ionotropic glutamate receptor (GluR1), as well as fragile X mental retardation protein (FMRP). The results support the conclusion that, surprisingly, increased RA signaling in the aged hippocampus is associated with poor cognitive outcome.


How normal is 'normal'? Mild tremor in a multiethnic cohort of normal subjects.
While many "normal" subjects exhibit mild clinically detectable tremor, the extent to which this tremor is present has received little attention. To characterize the prevalence and clinical characteristics of mild, clinically detectable tremor in a multiethnic cohort of normal subjects. Normal control subjects (n=36) and their relatives (n=67) were enrolled in a community-based case-control study of the familial aggregation of essential tremor. Subjects underwent a tremor interview and videotaped tremor examination. Two neurologists independently rated the severity of tremor during different postures and tasks, and a total tremor score (maximum score, 36) was calculated for each subject. Eight subjects were randomly selected to undergo quantitative computerized tremor analysis. In 103 normal subjects (36 control subjects and 67 relatives of control subjects) the mean total tremor score was 4.8 (range, 0-12.5). Ninety-nine (96%) of 103 subjects had tremor, as defined by a total tremor score of 0.5 or higher. For most tasks, the mean tremor score was greater in the nondominant than in the dominant arm. The total tremor score correlated highly with age (r=0.28; P=.004). There were no sex or ethnic differences in the mean total tremor scores. On tremor analysis, the amplitude and frequency of the tremor differed from that in a group of similarly studied subjects with essential tremor. Normal subjects almost uniformly have a clinically detectable tremor that is mild and age dependent. Characterization of this tremor helps to establish standards for normal tremor. These standards are crucial for accurate diagnostic classification in population-based studies of essential tremor.


Tailoring treatment of haemophilia B: accounting for the distribution and clearance of standard and extended half-life FIX concentrates.
The prophylactic administration of factor IX (FIX) is considered the most effective treatment for haemophilia B. The inter-individual variability and complexity of the pharmacokinetics (PK) of FIX, and the rarity of the disease have hampered identification of an optimal treatment regimens. The recent introduction of extended half-life recombinant FIX molecules (EHL-rFIX), has prompted a thorough reassessment of the clinical efficacy, PK and pharmacodynamics of plasma-derived and recombinant FIX. First, using longer sampling times and multi-compartmental PK models has led to more precise (and favourable) PK for FIX than was appreciated in the past. Second, investigating the distribution of FIX in the body beyond the vascular space (which is implied by its complex kinetics) has opened a new research field on the role for extravascular FIX. Third, measuring plasma levels of EHL-rFIX has shown that different aPTT reagents have different accuracy in measuring different FIX molecules. How will this new knowledge reflect on clinical practice? Clinical decision making in haemophilia B requires some caution and expertise. First, comparisons between different FIX molecules must be assessed taking into consideration the comparability of the populations studied and the PK models used. Second, individual PK estimates must rely on multi-compartmental models, and would benefit from adopting a population PK approach. Optimal sampling times need to be adapted to the prolonged half-life of the new EHL FIX products. Finally, costs considerations may apply, which is beyond the scope of this manuscript but might be deeply connected with the PK considerations discussed in this communication.


The incidence of congenital malformations in children with cancer.
We evaluated the incidence of congenital malformations in 566 children (median age: 8, M:F 1.3) with lymphomas and solid tumors using patient records. In this study, 12.7% of children either had a congenital malformation (7.8%) or a birthmark (4.9%). The incidence of patients with a childhood cancer syndrome was 3% and these cases developed typical tumors. The rate of consanguineous marriages was 12.6%, and family history of cancer was positive in 31.2%. Median age at cancer diagnosis, gender, maternal age, history of stillbirth and missed abortion, consanguinity of parents, and family history of cancer were not significantly different in cases with and without a congenital malformation. The most frequent cancers were central nervous system tumors and lymphomas. No remarkable association between a particular anomaly and a specific cancer type could be shown. The high incidence of congenital anomalies in this study may stimulate future large cohort studies in our country.


Laboratory and Neuroimaging Biomarkers in Neuropsychiatric Systemic Lupus Erythematosus: Where Do We Stand, Where To Go?
Systemic lupus erythematosus (SLE) is a chronic autoimmune disease characterized by multi-systemic involvement. Nervous system involvement in SLE leads to a series of uncommon and heterogeneous neuropsychiatric (NP) manifestations. Current knowledge on the underlying pathogenic processes and their subsequent pathophysiological changes leading to NP-SLE manifestations is incomplete. Several putative laboratory biomarkers have been proposed as contributors to the genesis of SLE-related nervous system damage. Alongside the laboratory biomarkers, several neuroimaging tools have shown to reflect the nature of tissue microstructural damage associated with SLE, and thus were suggested to contribute to the understanding of the pathophysiological changes and subsequently help in clinical decision making. However, the number of useful biomarkers in NP-SLE in clinical practice is disconcertingly modest. In some cases it is not clear whether the biomarker is truly involved in pathogenesis, or the result of non-specific pathophysiological changes in the nervous system (e.g., neuroinflammation) or whether it is the consequence of a concomitant underlying abnormality related to SLE activity. In order to improve the diagnosis of NP-SLE and provide a better targeted care to these patients, there is still a need to develop and validate a range of biomarkers that reliably capture the different aspects of disease heterogeneity. This article critically reviews the current state of knowledge on laboratory and neuroimaging biomarkers in NP-SLE, discusses the factors that need to be addressed to make these biomarkers suitable for clinical application, and suggests potential future research paths to address important unmet needs in the NP-SLE field.


Identification of α-tocopherol and α-tocopheryl acetate from the cuticle of soybean pods armyworm (Spodoptera cosmioides).
The chemical composition of the soybean pods armyworm Spodoptera cosmioides (Walker, 1858) (Lepidoptera: Noctuidae) and Anticarsia gemmatalis Hübner, 1818 (Lepidoptera: Erebidae) larval cuticles was evaluated using gas chromatography coupled to a mass detector (GC-MS). Among the usual lipids found in the insect cuticle, α-tocopherol and α-tocopheryl acetate were also isolated from S. cosmioides. On the other hand, no vitamin E derivative was found in A. gemmatalis exuvia. This is the first report of vitamin E occurrence in the insect's cuticle.


Unique Chemokine Profiles of Lung Tissues Distinguish Post-chemotherapeutic Persistent and Chronic Tuberculosis in a Mouse Model.
There is a substantial need for biomarkers to distinguish latent stage from active Mycobacterium tuberculosis infections, for predicting disease progression. To induce the reactivation of tuberculosis, we present a new experimental animal model modified based on the previous model established by our group. In the new model, the reactivation of tuberculosis is induced without administration of immunosuppressive agents, which might disturb immune responses. To identify the immunological status of the persistent and chronic stages, we analyzed immunological genes in lung tissues from mice infected with M. tuberculosis. Gene expression was screened using cDNA microarray analysis and confirmed by quantitative RT-PCR. Based on the cDNA microarray results, 11 candidate cytokines genes, which were obviously up-regulated during the chronic stage compared with those during the persistent stage, were selected and clustered into three groups: (1) chemokine genes, except those of monocyte chemoattractant proteins (MCPs; CXCL9, CXCL10, CXCL11, CCL5, CCL19); (2) MCP genes (CCL2, CCL7, CCL8, CCL12); and (3) TNF and IFN-γ genes. Results from the cDNA microarray and quantitative RT-PCR analyses revealed that the mRNA expression of the selected cytokine genes was significantly higher in lung tissues of the chronic stage than of the persistent stage. Three chemokines (CCL5, CCL19, and CXCL9) and three MCPs (CCL7, CCL2, and CCL12) were noticeably increased in the chronic stage compared with the persistent stage by cDNA microarray (p < 0.01, except CCL12) or RT-PCR (p < 0.01). Therefore, these six significantly increased cytokines in lung tissue from the mouse tuberculosis model might be candidates for biomarkers to distinguish the two disease stages. This information can be combined with already reported potential biomarkers to construct a network of more efficient tuberculosis markers.