GM biofortified crops: potential effects on targeting the micronutrient intake gap in human populations.
Genetic engineering has been successfully applied to increase micronutrient content in staple crops. Nutrition evidence is key to ensure scale-up and successful implementation. Unlike conventional plant breeding efforts, research on the efficacy or effectiveness of GM biofortified crops on nutritional status in human populations is lacking. This review reports on the potential role of GM biofortified crops in closing the micronutrient gap - increasing the dietary intake of micronutrients in human populations. To date, one clinical trial in the United States reported a high bio-conversion rate of β-carotene in Golden Rice, and potential effects of GM biofortified crop consumption on dietary intake and nutritional outcomes are promising. However, further research needs to confirm the ex ante assessments in target regions.


Determination of gene copy number and genotype of transgenic Arabidopsis thaliana by competitive PCR.
A simple and rapid method is described for determining the integrated T-DNA copy number and the genotype in transgenic Arabidopsis thaliana by two-step competitive PCR. First, the amount of genomic DNA in the extracts, obtained from an individual A. thaliana transformant, was accurately determined by the 1st competitive PCR using a known single copy gene, 4HPPD (4-hydroxyphenylpyruvate dioxygenase), as a target. Second, the number of T-DNA copies per genome was estimated by quantifying the NPTII gene, which was involved in the T-DNA, by the 2nd competitive PCR using exactly the same amount of genomic DNA for each sample. The estimated copy number and genotype obtained by this procedure were identical to those determined by Southern blot analysis and segregation analysis.


Growth during puberty in cystic fibrosis: a retrospective evaluation of a French cohort.
To assess the longitudinal growth pattern in a large French cohort of patients with cystic fibrosis (CF), to determine to what extent puberty contributed to final height and to explore a potential relationship between growth, nutritional status and respiratory function. Retrospective data were drawn from the French CF registry from 1999 to 2004. Height, weight and forced expiratory volume in 1 s (FEV(1)) were recorded annually. Growth and velocity curves were compared with reference curves. 729 children with CF were included. In girls, height was similar to the reference population until age 11. Age at onset of puberty was the same as in reference girls. The pubertal spurt was lower than reference values and contributed less to the final adult height. In boys, the mean height was close to the reference mean until age 14 and was thereafter lower. Age at growth acceleration was similar to that in reference boys, but with an impaired peak height velocity (PHV). The final height was lower than in the general population (z score -0.73). No correlations were found between body mass index (BMI) and PHV. In girls, there was a weak but significant positive relationship between PHV and FEV(1) (r=0.17, p=0.02) In this cohort, children with CF had a normal age of onset of puberty and pubertal spurt, with a low PHV. Puberty contributed less to final adult height, and neither BMI nor FEV(1) had a significant effect on pubertal height gain.


Apolipoprotein A5 gene promoter region-1131T/C polymorphism is associated with risk of ischemic stroke and elevated triglyceride levels: a meta-analysis.
The association between polymorphism -1131T/C in the promoter region of apolipoprotein A5 (APOA5) and ischemic stroke and plasma triglyceride (TG) levels remains controversial. To better clarify the association between APOA5-1131T/C and risk of ischemic stroke and plasma TG levels, we performed a meta-analysis to examine the allele and genotype of APOA5-1131T/C polymorphism in ischemic stroke cases and controls. Based on the search of PubMed, Embase, MEDLINE, CNKI (National Knowledge Infrastructure) and CBM (Chinese BioMedical Literature Database) databases, we identified and abstracted outcome data from all articles to evaluate the association between APOA5 and ischemic stroke/plasma TG levels. The pooled odds ratios (ORs) with 95% confidence intervals (CIs) were performed in dominant model (CC + TC vs. TT), recessive model (CC vs. TC + TT), homozygote comparison (CC vs. TT) and heterozygote comparison (TC vs. TT). The association between dominant model (CC + TC vs. TT) and plasma TG/total cholesterol/high-density lipoprotein cholesterol levels was measured by a weighted mean difference (WMD) with its corresponding 95% CI. To evaluate the ethnicity-specific effects, subgroup analyses were performed by ethnic group. A meta-analysis containing 2,294 ischemic stroke cases and 1,858 controls from 8 case-control studies was performed. The results showed that APOA5-1131T/C polymorphism was significantly associated with ischemic stroke in all comparison models (CC + TC vs. TT, OR = 1.70, 95% CI = 1.24-2.32; CC vs. TC + TT, OR = 1.36, 95% CI = 0.98-1.90; CC vs. TT, OR = 1.73, 95% CI = 1.34-2.23; TC vs. TT, OR = 1.67, 95% CI = 1.19-2.36). On subgroup analysis by ethnicity, similarly significant associations were found in both Asians and Europeans, and the Europeans possessed a higher risk of ischemic stroke, especially in CC versus TT model (OR = 4.47, 95% CI = 1.33-15.06). Significant association between the C allele and elevated TG levels was detected in both ischemic stroke cases and controls; the TG levels were higher in the ischemic stroke cases and controls carrying the APOA5-1131C allele than in the noncarriers (CC + TC vs. TT, cases WMD = 0.43, 95% CI = 0.27-0.59; controls WMD = 0.51, 95% CI = 0.35-0.66). Similar within-group comparison of the total cholesterol and high-density lipoprotein cholesterol levels did not show any difference. Our meta-analysis revealed that the APOA5-1131T/C polymorphism is associated with a significant risk of ischemic stroke and elevated TG levels. The CC genotype and C allele might be a genetic risk factor that increases susceptibility of ischemic stroke and elevates plasma TG levels, and might be a useful target for clinical therapeutic intervention.


Supramolecular Assembly of Ag(I) Centers: Diverse Topologies Directed by Anionic Interactions.
Ag(I)-Ag(I) interactions in supramolecular structures have been achieved through the use of structural support from the ligand frames. In structures involving simple ligands like pyridine, strong π-π interaction leads to spatial ordering of the individual [Ag(L)2]+ units. In such structures anions also play a crucial role in dictating the final arrangement of the [Ag(L)2]+ synthons. In order to determine whether the anions can solely dictate the arrangement of the [Ag(L)2]+ synthons in the supramolecular structure, four Ag(I) complexes of 4-pyridylcarbinol (PyOH), namely, [Ag(PyOH)2]X (X = NO3- (1), BF4- (2), CF3SO3- (3), and ClO4- (4)) have been synthesized and structurally characterized. Gradual transformation of the extended structures observed in 1-3 eventually merges into a unique linear alignment of the [Ag(PyOH)2]+ units in 4 along the c axis, a feature that results in strong argentophilic interactions. Complex 4 is sensitive to light and is inherently less stable than the other three analogues. The structural variations in this set of extended assemblies are solely dictated by the anions, since π-π interaction between the substituted pyridine ligands is significantly diminished due to disposition of the -CH2OH substituent at the 4 position and H-bonding throughout the structure.


Functional implications of a psychiatric risk variant within CACNA1C in induced human neurons.
Psychiatric disorders have clear heritable risk. Several large-scale genome-wide association studies have revealed a strong association between susceptibility for psychiatric disorders, including bipolar disease, schizophrenia and major depression, and a haplotype located in an intronic region of the L-type voltage-gated calcium channel (VGCC) subunit gene CACNA1C (peak associated SNP rs1006737), making it one of the most replicable and consistent associations in psychiatric genetics. In the current study, we used induced human neurons to reveal a functional phenotype associated with this psychiatric risk variant. We generated induced human neurons, or iN cells, from more than 20 individuals harboring homozygous risk genotypes, heterozygous or homozygous non-risk genotypes at the rs1006737 locus. Using these iNs, we performed electrophysiology and quantitative PCR experiments that demonstrated increased L-type VGCC current density as well as increased mRNA expression of CACNA1C in iNs homozygous for the risk genotype, compared with non-risk genotypes. These studies demonstrate that the risk genotype at rs1006737 is associated with significant functional alterations in human iNs, and may direct future efforts at developing novel therapeutics for the treatment of psychiatric disease.


Modulating Ocular Surface Pain Through Neurokinin-1 Receptor Blockade.
The purpose of this study was to test the role of substance P (SP) and its receptor neurokinin 1 (NK1R) on ocular surface pain. Eight-week-old C57BL6/N (wild type [WT]) and B6.Cg-Tac1tm1Bbm/J (TAC1-KO) male mice were used. 5 M NaCl was topically applied on the cornea, followed by topical fosaprepitant 2, 10, and 50 mg/mL; 4 mg/mL oxybuprocaine chloride, or 0.1% diclofenac. Th eye wiping test was used to quantify ocular surface pain. SP content was quantified in the tear fluid and trigeminal ganglia (TG), and TAC1 mRNA was assessed in the cornea. Corneas were immunostained for β3-tubulin and NK1R, or CD45, to quantify leukocyte infiltration. TAC1-KO mice displayed a significant reduction of ocular pain (P < 0.001). Similarly, a single dose of 10 or 50 mg/mL fosaprepitant applied topically to WT mice reduced ocular pain as compared to vehicle (P < 0.001). Fosaprepitant 2 mg/mL, instead, induced corneal analgesia only when it was administered for 10 days, 6 times/day (P < 0.05). Diclofenac or oxybuprocaine reduced corneal nociception when compared to vehicle or fosaprepitant (P < 0.05). Fosaprepitant or oxybuprocaine groups showed lower SP content in tear secretions and TG (P < 0.05), and reduction in TAC1 mRNA (P < 0.05), and leukocyte infiltration (P < 0.05) in the cornea. Colocalization of NK1R and β3-tubulin was detected in mouse corneas. Topical administration of the NK1R antagonist fosaprepitant effectively reduces ocular surface nociception by decreasing SP release in the tear fluid and TG, and corneal leukocyte infiltration. Fosaprepitant repurposing shows promise for the treatment of ocular pain.


HOW MUCH HERITABLE VARIATION CAN BE MAINTAINED IN FINITE POPULATIONS BY MUTATION-SELECTION BALANCE?
The joint effects of stabilizing selection, mutation, recombination, and random drift on the genetic variability of a polygenic character in a finite population are investigated. A simulation study is performed to test the validity of various analytical predictions on the equilibrium genetic variance. A new formula for the expected equilibrium variance is derived that approximates the observed equilibrium variance very closely for all parameter combinations we have tested. The computer model simulates the continuum-of-alleles model of Crow and Kimura. However, it is completely stochastic in the sense that it models evolution as a Markov process and does not use any deterministic evolution equations. The theoretical results are compared with heritability estimates from laboratory and natural populations. Heritabilities ranging from 20% to 50%, as observed even in lab populations under a constant environment, can only be explained by a mutation-selection balance if the phenotypic character is neutral or the number of genes contributing to the trait is sufficiently high, typically several hundred, or if there are a few highly variable loci that influence quantitative traits.


Raw deal for workers: why have U.S. workers experienced a long-term decline in pay, benefits, and working conditions?
Pay, opportunities, and job quality have worsened for most U.S. workers over the past 30 years, across most sectors of the economy. This decline is related to fundamental changes in the economy and society, including sluggish productivity growth and employer assaults on workers' rights and protections. Productivity growth has slowed as companies no longer invest as much in equipment and training. Businesses have attacked workers' protections; unionization is down, and the minimum wage is worth about two-thirds as much as at its high point in the late 1960s. The National Labor Relations Board, other federal agencies, and the courts are stacked with anti-labor appointees. And businesses have pushed more and more risk onto workers, with a growth in temporary work and much reduced work-related benefits.


Stereoselective Access to Alkylidenecyclobutanes through γ-Selective Cross-Coupling Strategies.
Alkylidenecyclobutanes (ACBs) containing all-carbon quaternary stereocenters were simply and efficiently synthesized by combining boron-homologation and γ-selective cross-coupling strategies. This unique sequence led to excellent regio- and diastereoselectivities in the generation of targeted four-membered rings with up to 99% enantiomeric excess using chiral substrates. In addition to the original synthesis of ACBs, the first asymmetric catalytic formation of quaternary stereocenters based on γ-selective cross-coupling reactions is finally shown.


Cardiotrophin-like cytokine (CLCF1) modulates mesenchymal stem cell osteoblastic differentiation.
Mesenchymal stem cells (MSCs) are multipotent cells capable of differentiating into adipocytes, chondrocytes, or osteocytes. MSCs secrete an array of cytokines and express the LIFRβ (leukemia inhibitory factor receptor) chain on their surface. Mutations in the gene coding for LIFRβ lead to a syndrome with altered bone metabolism. LIFRβ is one of the signaling receptor chains for cardiotrophin-like cytokine (CLCF1), a neurotrophic factor known to modulate B and myeloid cell functions. We investigated its effect on MSCs induced to differentiate into osteocytes in vitro Our results indicate that CLCF1 binds mouse MSCs, triggers STAT1 and -3 phosphorylation, inhibits the up-regulation of master genes involved in the control of osteogenesis, and markedly prevents osteoblast generation and mineralization. This suggests that CLCF1 could be a target for therapeutic intervention with agents such as cytokine traps or blocking mAbs in bone diseases such as osteoporosis.


Heterogeneous uptake of ozone on reactive components of mineral dust aerosol: an environmental aerosol reaction chamber study.
We have undertaken a kinetic study of heterogeneous ozone decomposition on alpha-Fe2O3 (hematite) and alpha-Al2O3 (corundum) aerosols under ambient conditions of temperature, pressure, and relative humidity in order to better understand the role of mineral dust aerosol in ozone loss mechanisms in the atmosphere. The kinetic measurements are made in an environmental aerosol reaction chamber by use of infrared and ultraviolet spectroscopic probes. The apparent heterogeneous uptake coefficient, gamma, for ozone reaction with alpha-Fe2O3 and alpha-Al2O3 surfaces is determined as a function of relative humidity (RH). The uptake of ozone by the iron oxide surface is approximately an order of magnitude larger than that by the aluminum oxide sample, under dry conditions. At the pressures used, alpha-Fe2O3 shows clear evidence for catalytic decomposition of ozone while alpha-Al2O3 appears to saturate at a finite ozone coverage. The measured uptake for both minerals decreases markedly as the RH is increased. Comparison with other literature reports and the atmospheric implications of these results are discussed.


Primary CNS germ cell tumors: current epidemiology and update on treatment.
Primary central nervous system (CNS) germ cell tumors (GCTs) are a heterogeneous group of lesions that account for 0.5 % of all primary brain and CNS tumors, occurring at an incidence rate of 0.10 per 100,000 person-years in the United States with approximately 90 % of the cases before the age of 20 years. Primary CNS GCTs demonstrate a remarkable difference in incidence based on gender and location within the brain with males having a 15:1 incidence in the pineal region while the gender incidence is nearly 1:1 in the rest of the brain. Also, historically the incidence was noted to be significantly higher in Japan and East Asia, but recent studies in Japan demonstrate similar incidence as in the United States. They are broadly classified as germinomas and non-germinomatous germ cell tumors (NGGCTs) based on clinicopathologic features. Germinomas are sensitive to treatment with radiotherapy and chemotherapy with high cure rates and carry an excellent prognosis, while NGGCTs display various forms of differentiation, have a poorer prognosis and are refractory to therapy. Standard management of CNS GTCs remains unsettled and ongoing research aims to achieve best possible survival rates and post-treatment quality of life by reduction in treatment intensity.


Incidence of factor FVIIIC deficiency in live male infants undergoing circumcision in South West, Nigeria.
A prominent evidence of inherited bleeding disorder in newborn males is excessive post-circumcision bleeding. Male circumcision in Nigeria is the rule rather than the exception. Male siblings of some of the Nigerian haemophiliacs consequently died from severe post-circumcision bleeding. The aim was to determine the incidence of inherited factor VIIIc (FVIIIc) deficiencies in live male infants undergoing circumcision in South West, Nigeria. The study population was 244 male infants drawn from University College Hospital and Our Lady of Apostles Catholic Hospital, Oluyoro, Ibadan. Pre-circumcision prothrombin time, activated partial thromboplastin time and FVIIIc levels were determined. Clinical features of inherited bleeding disorder particularly family history of bleeding diathesis, history of cephalhaematoma and bleeding from the umbilical stump in neonatal life were determined with the aid of a questionnaire. Only one of the mothers (0.4%) gave a family history of bleeding disorder. A history of excessive bleeding from the umbilical stump post delivery was obtained in three (2%) of the patients. Five (2%) other subjects had cephalhaematoma post delivery. Two of the subjects (0.8%) had prolonged activated partial thromboplastin time. The factor VIIIc level was between 31% and 49% in 16.1%, while 1.6% of the neonates had levels between 20% and 26%. This study detected four of the 244 (1.64%) neonates with FVIIIc deficiency, suggestive of either mild haemophilia or von Willebrand's disease. A larger study (including family studies) will be required, so as to arrive at the exact incidence of both haemophilia A and vWD in live male infants in Nigeria.


Extracellular Vesicle Delivery of TRAIL Eradicates Resistant Tumor Growth in Combination with CDK Inhibition by Dinaciclib.
Tumour necrosis factor (TNF)-related apoptosis inducing ligand (TRAIL) is a promising anti-cancer agent that rapidly induces apoptosis in cancer cells. Unfortunately, the clinical application of recombinant TRAIL (rTRAIL) has been hampered by its common cancer resistance. Naturally TRAIL is delivered as a membrane-bound form by extracellular vesicles (EV-T) and is highly efficient for apoptosis induction. SCH727965 (dinaciclib), a potent cyclin-dependent kinase (CDK) inhibitor, was shown to synergize with other drugs to get better efficacy. However, it has never been investigated if dinaciclib coordinates with EV-T to enhance therapeutic results. This study explores the potential of combination therapy with EV-T and dinaciclib for cancer treatment. EV-T was successfully derived from human TRAIL transduced cells and shown to partially overcome resistance of A549 cells. Dinaciclib was shown to drastically enhance EV-T killing effects on cancer lines that express good levels of death receptor (DR) 5, which are associated with suppression of CDK1, CDK9 and anti-apoptotic proteins. Combination therapy with low doses of EV-T and dinaciclib induced strikingly enhanced apoptosis and led to complete regression in A549 tumors without any adverse side effects observed in a subcutaneous xenograft model. Tumor infiltration of mass NK cells and macrophages was also observed. These observations thus indicate that the combination of EV-T with dinaciclib is a potential novel therapy for highly effective and safe cancer treatment.


Regional contributions of six preventable risk factors to achieving the 25 × 25 non-communicable disease mortality reduction target: a modelling study.
Countries have agreed to reduce premature mortality from the four main non-communicable diseases (NCDs) by 25% from 2010 levels by 2025 (referred to as the 25 × 25 target). Countries also agreed on a set of global voluntary targets for selected NCD risk factors. Previous analyses have shown that achieving the risk factor targets can contribute substantially towards meeting the 25 × 25 mortality target at the global level. We estimated the contribution of achieving six of the globally agreed risk factor targets towards meeting the 25 × 25 mortality target by region. We estimated the effect of achieving the targets for six risk factors (tobacco and alcohol use, salt intake, obesity, and raised blood pressure and glucose) on NCD mortality between 2010 and 2025. Our methods accounted for multicausality of NCDs and for the fact that, when risk factor exposure increases or decreases, the harmful or beneficial effects on NCDs accumulate gradually. We used data for risk factor and mortality trends from systematic analyses of available country data. Relative risks for the effects of individual and multiple risks, and for change in risk after decreases or increases in exposure, were from reanalyses and meta-analyses of epidemiological studies. The probability of dying between the ages 30 years and 70 years from the four main NCDs in 2010 ranged from 19% in the region of the Americas to 29% in southeast Asia for men, and from 13% in Europe to 21% in southeast Asia for women. If current trends continue, the probability of dying prematurely from the four main NCDs is projected to increase in the African region but decrease in the other five regions. If the risk factor targets are achieved, the 25 × 25 target will be surpassed in Europe in both men and women, and will be achieved in women (and almost achieved in men) in the western Pacific; the regions of the Americas, the eastern Mediterranean, and southeast Asia will approach the target; and the rising trend in Africa will be reversed. In most regions, a more ambitious approach to tobacco control (50% reduction relative to 2010 instead of the agreed 30%) will contribute the most to reducing premature NCD mortality among men, followed by addressing raised blood pressure and the agreed tobacco target. For women, the highest contributing risk factor towards the premature NCD mortality target will be raised blood pressure in every region except Europe and the Americas, where the ambitious (but not agreed) tobacco reduction would have the largest benefit. No WHO region will meet the 25 × 25 premature mortality target if current mortality trends continue. Achieving the agreed targets for the six risk factors will allow some regions to meet the 25 × 25 target and others to approach it. Meeting the 25 × 25 target in Africa needs other interventions, including those addressing infection-related cancers and cardiovascular disease. UK Medical Research Council.


New insights into functions of the sphingosine-1-phosphate transporter SPNS2.
Sphingosine-1-phosphate (S1P) is a potent bioactive signaling molecule that regulates many physiological processes important for development, epithelial and endothelial barrier integrity, and the immune system, as well as for pathologies, such as autoimmune diseases, cancer, and metastasis. Most of the well-known actions of S1P are mediated by five specific G protein-coupled receptors located on the plasma membrane. Because S1P is synthesized intracellularly by two sphingosine kinase isoenzymes, we have proposed the paradigm of inside-out signaling by S1P, suggesting that S1P must be exported out of cells to interact with its receptors. While several transporters of S1P have previously been identified, spinster homologue 2 (SPNS2), a member of the large family of non-ATP-dependent organic ion transporters, has recently attracted much attention as an S1P transporter. Here, we discuss recent advances in understanding the physiological actions of SPNS2 in regulating levels of S1P and the S1P gradient that exists between the high circulating concentrations of S1P and low tissue levels that control lymphocyte trafficking. Special emphasis is on the functions of SPNS2 in inflammatory and autoimmune diseases and its recently discovered unexpected importance in metastasis.


Microbubble-mediated sonothrombolysis with BR38 of a venous full blood thrombus in a rat embolic stroke model.
Early recanalization of an occluded vessel is associated with a better clinical outcome in acute ischemic stroke. Intravenous thrombolysis using recombinant tissue plasminogen activator (rt-PA) is only available in a minority of patients and often fails to reopen the occluded vessel. Mechanical recanalization is more effective in this matter but only available for selected patients when a thrombectomy centre can be reached. Therefore, sonothrombolysis might represent an alternative or complementary approach. Here, we tested microbubble-mediated sonothrombolysis (mmSTL) in a thromboembolic stroke model for middle cerebral artery occlusion (MCAO) in rats. Sixty-seven male Wistar rats underwent MCAO using an autologous full blood thrombus and were randomly assigned to four groups receiving rt-PA, mmSTL, a combination of both, or a placebo. Diagnostic workup included neurological examination, assessment of infarct size, and presence of intracerebral haemorrhage by magnetic resonance imaging (MRI) and presence of microbleedings in histological staining. Neurological examination revealed no differences between the treatment groups. In all treatment groups, there was a reduction in infarct size 24 hours after MCAO as compared to the placebo (P≤0.05), but there were no differences between the active treatment groups (P>0.05) (placebo 0.75±0.10 cm3; mmSTL 0.43±0.07 cm3; rt-PA 0.4±0.07 cm3; mmSTL + rt-PA 0.27±0.08 cm3). Histological staining displayed intracerebral microbleedings in all animals. The frequency of gross bleeding detected by MRI did not differ between the groups (placebo 3; mmSTL 4; rt-PA 2; mmSTL + rt-PA 2; P>0.05) and was not associated with worse performance in clinical testing (P>0.05). There were no statistical differences in the mortality between the groups (P>0.05). Our study showed the efficacy and safety of mmSTL with or without rt-PA in an embolic rat stroke model using a continuous full blood thrombus. Sonothrombolysis might be useful for patients who need to be transported to a thrombectomy centre or for those with distal vessel occlusion.


Comparative analysis of E2F family member oncogenic activity.
The E2F family of transcription factors consists of nine members with both distinct and overlapping functions. These factors are situated downstream of growth factor signaling cascades, where they play a central role in cell growth and proliferation through their ability to regulate genes involved in cell cycle progression. For this reason, it is likely that the members of the E2F family play a critical role during oncogenesis. Consistent with this idea is the observation that some tumors exhibit deregulated expression of E2F proteins. In order to systematically compare the oncogenic capacity of these family members, we stably over-expressed E2F1 through 6 in non-transformed 3T3 fibroblasts and assessed the ability of these transgenic cell lines to grow under conditions of low serum, as well as to form colonies in soft agar. Our results show that these six E2F family members can be divided into three groups that exhibit differential oncogenic capacity. The first group consists of E2F2 and E2F3a, both of which have strong oncogenic capacity. The second group consists of E2F1 and E2F6, which were neutral in our assays when compared to control cells transduced with vector alone. The third group consists of E2F4 and E2F5, which generally act to repress E2F-responsive genes, and in our assays demonstrated a strong capacity to inhibit transformation. Our results imply that the pattern of expression of these six E2F family members in a cell could exert a strong influence over its susceptibility to oncogenic transformation.


[Physical activity programs that include self-efficacy in school children with obesity: a systematic review].
Introduction: this research aims to carry out a systematic review of physical activity programs that improve self-efficacy in children and adolescents with obesity, to realize their characteristics and thus design a more comprehensive and effective program for this population. Objective: to analyze the characteristics of physical activity programs that improve self-efficacy in children and adolescents with obesity, through a systematic review. Methodology: systematic review of observational-descriptive design; the search was realized in databases of the health area: PubMed, ERIC, Cochrane, Redalyc, and Google academic. Free access articles in English and Spanish from the last 10 years were selected. The temporality was from March 2012 to May 2022. The descriptors used were; "physical activity program", "training program", "self-efficacy", "children", "adolescents", and "obesity". Results: six studies were included (n = 10). Different types of interventions that improved self-efficacy in the participant were identified, such as recreational activities, traditional games, sports, and structured activities for health care. The methodology implemented and the timing and duration were key variables to show positive changes in childhood overweight and obesity (6 a 19 years). Conclusions: it is crucial to identify the principal health care characteristics included in programs focused on increasing self-efficacy in childhood. These features lay the foundation for designing comprehensive programs.


Environmental enrichment attenuates hippocampal neuroinflammation and improves cognitive function during influenza infection.
Recent findings from our lab have shown that peripheral infection of adult mice with influenza A/PR/8/34 (H1N1) virus induces a neuroinflammatory response that is paralleled by loss of neurotrophic and glial regulatory factors in the hippocampus, and deficits in cognitive function. Environmental enrichment has been shown to exert beneficial effects on the brain and behavior in many central nervous system (CNS) disorders, but its therapeutic potential during peripheral viral infection remains unknown. Therefore, the objective of the present study was to determine if long-term continuous exposure to environmental enrichment could prevent and/or attenuate the negative effects of influenza infection on the hippocampus and spatial cognition. Mice were housed in enriched or standard conditions for 4 months, and continued to live in their respective environments throughout influenza infection. Cognitive function was assessed in a reversal learning version of the Morris water maze, and changes in hippocampal expression of proinflammatory cytokines (IL-1β, IL-6, TNF-α, IFN-α), neurotrophic (BDNF, NGF), and immunomodulatory (CD200, CX3CL1) factors were determined. We found that environmental enrichment reduced neuroinflammation and helped prevent the influenza-induced reduction in hippocampal CD200. These changes were paralleled by improved cognitive performance of enriched mice infected with influenza when compared to infected mice in standard housing conditions. Collectively, these data are the first to demonstrate the positive impact of environmental enrichment on the brain and cognition during peripheral viral infection, and suggest that enhanced modulation of the neuroimmune response may underlie these beneficial effects.


Combined treatment of dysfunctional dialysis access with cutting balloon and paclitaxel-coated balloon in real world.
Background: Dysfunction in arteriovenous (AV) access is a common reason for subsequent intervention. To evaluate the results of endovascular treatment of AV access lesions using cutting balloon (CB) and drug-coated balloon (DCB) angioplasty compared to standard treatment using plain-old balloon angioplasty (POBA). Patients and methods: Patients who retrospectively were selected from a prospectively maintained database. Primary endpoint was primary patency rate at the target lesion up to 12 months after index procedure. Secondary endpoints were the acute treatment success, the "bail out" stent rate, primary patency at 6 months, freedom from target lesion revascularization (TLR), AV access replacement and surgical revision during a follow-up period up to 12 months, and all-cause mortality rate stratified to patients treated with and without DCB. Results: One hundred and eighty-four patients met the inclusion criteria. POBA as stand alone or combined with DCB angioplasty was performed in 71 patients (38.6%), CB in 54 patients (29.3%), and in 59 patients (32.1%), both CB and DCB were used. Primary patency rate at 12 months was 31.6% for the POBA/DCB-group, 52.3% for the CB-group, and 64.8% for the CB/DCB-group, respectively. In total, 80 patients (51.6%) had a TLR including endovascular or surgical revision, or a shunt replacement. All-cause mortality at 12 months was 7.2% in the DCB group and 9.1% in the group of patients treated without a DCB (p=0.747). Conclusions: The use of CB seems to be crucial for a better outcome. The combination of CB and DCB achieves the best patency results at mid-term.


Snack bar compositions and their acute glycaemic and satiety effects.
Maintaining blood glucose within homeostatic limits and eating foods that sup-press hunger and promote satiety have beneficial impacts for health. This study investigated the glycaemic re-sponse and satiety effects of a serving size of a healthier snack bar, branded Nothing Else, that met the required nutrient profiling score criteria for a health claim, in comparison to two top-selling commercial snack bars. In an experimental study, 24 participants aged >=50 years were recruited. On three different days blood glucose concentration was measured twice at baseline and 15, 30, 45, 60, 90 and 120 minutes after consumption of a serving size of each bar. Satiety effects were self-reported hunger, fullness, desire to eat, and amount could eat ratings on visual analogue scales. The incremental area under the blood glucose response curve (iAUC) over two hours for the Nothing Else bar was 30% lower than commercial Bar 2 (p<0.001). At 45 minutes after eating, the Nothing Else bar induced the highest fullness rating and lowest hunger rating among the three snack bars. At two hours, fullness induced by the Nothing Else bar was twice that of Bar 2 (p=0.019), but not different to Bar 1 (p=0.212). The Nothing Else snack bar developed using the nutrient profiling scheme as a guideline, with its high protein and dietary fibre contents, had a lower glycaemic impact and induced a higher subjective satiety than the two commercial snack bars of equal weight.


Changes in microvascular density differentiate metabolic health outcomes in monkeys with prior radiation exposure and subsequent skeletal muscle ECM remodeling.
Radiation exposure accelerates the onset of age-related diseases such as diabetes, cardiovascular disease, and neoplasia and, thus, lends insight into in vivo mechanisms common to these disorders. Fibrosis and extracellular matrix (ECM) remodeling, which occur with aging and overnutrition and following irradiation, are risk factors for development of type 2 diabetes mellitus. We previously demonstrated an increased incidence of skeletal muscle insulin resistance and type 2 diabetes mellitus in monkeys that had been exposed to whole body irradiation 5-9 yr prior. We hypothesized that irradiation-induced fibrosis alters muscle architecture, predisposing irradiated animals to insulin resistance and overt diabetes. Rhesus macaques (Macaca mulatta, n = 7-8/group) grouped as nonirradiated age-matched controls (Non-Rad-CTL), irradiated nondiabetic monkeys (Rad-CTL), and irradiated monkeys that subsequently developed diabetes (Rad-DM) were compared. Prior radiation exposure resulted in persistent skeletal muscle ECM changes, including a relative overabundance of collagen IV and a trend toward increased transforming growth factor-β1. Preservation of microvascular markers differentiated the irradiated diabetic and nondiabetic groups. Microvascular density and plasma nitrate and heat shock protein 90 levels were lower in Rad-DM than Rad-CTL. These results are consistent with a protective effect of abundant microvasculature in maintaining glycemic control within radiation-induced fibrotic muscle.


Physician specialty and variations in adoption of electronic health records.
Efforts to promote adoption of electronic health records (EHRs) have focused on primary care physicians, who are now expected to exchange data electronically with other providers, including specialists. However, the variation of EHR adoption among specialists is underexplored. We conducted a retrospective cross-sectional study to determine the association between physician specialty and the prevalence of EHR adoption, and a retrospective serial cross-sectional study to determine the association of physician specialty and the rate of EHR adoption over time. We used the 2005-2009 National Ambulatory Medical Care Survey. We considered fourteen specialties, and four definitions of EHR adoption (any EHR, basic EHR, full EHR, and a novel definition of EHR sophistication). We used multivariable logistic regression, and adjusted for several covariates (geography, practice characteristics, revenue characteristics, physician degree). Physician specialty was significantly associated with EHR adoption, regardless of the EHR definition, after adjusting for covariates. Psychiatrists, dermatologists, pediatricians, ophthalmologists, and general surgeons were significantly less likely to adopt EHRs, compared to the reference group of family medicine/general practitioners. After adjustment for covariates, these specialties were 44-94% less likely to adopt EHRs than the reference group. EHR adoption increased in all specialties, by approximately 40% per year. The rate of EHR adoption over time did not significantly vary by specialty. Although EHR adoption is increasing in all specialties, adoption varies widely by specialty. In order to insure each individual's network of providers can electronically share data, widespread adoption of EHRs is needed across all specialties.


Inferring diet, feeding behaviour and causes of mortality from prey-induced injuries in a New Zealand fur seal.
New Zealand fur seals Arctocephalus forsteri are the most abundant of the 4 otariid (eared seal) species distributed across Australasia. Analyses of stomach contents, scats and regurgitates suggest a diet dominated by bony fish and squid, with cartilaginous species (e.g. sharks and rays) either absent or underrepresented because of a lack of preservable hard parts. Here we report on a subadult specimen from south-eastern Australia, which was found ashore emaciated and with numerous puncture wounds across its lips, cheeks, throat and the inside of its oral cavity. Fish spines embedded in the carcass revealed that these injuries were inflicted by chimaeras and myliobatiform rays (stingrays and relatives), which matches reports on the diet of A. forsteri from New Zealand, but not South Australia. Shaking and tearing of prey at the surface may help to avoid ingestion of the venomous spines, perhaps contributing to their absence from scats and regurgitates. Nevertheless, the number and severity of the facial stab wounds, some of which led to local necrosis, likely affected the animal's ability to feed, and may account for its death. Despite their detrimental effects, fish spine-related injuries are difficult to spot, and may be a common, albeit cryptic, type of trauma. We therefore recommend that stranded seals be systematically examined for this potentially life-threatening pathology.


Reconstruction of the Exenterated Orbit with an Island Pericranial Flap: A New Surgical Approach.
Reconstruction of the bony socket after orbital exenteration is a matter of much debate. Prompt defect closure with a microvascular flap is desirable but involves a major surgical procedure and hence, places considerable burden on the patient. The new surgical technique presented here permits a technically simpler wound closure with fewer complications after orbital exenteration. Between May 2014 and June 2022 in the ENT department of Regensburg University, nine patients underwent exenteration and reconstruction with a pericranial flap. The flap was raised via a broken line incision in the forehead or endoscopically, incised in a roughly croissant-like shape, then introduced into the orbit through a tunnel in the eyebrow. A retrospective analysis of the patients and considerations about determining the size, shape, and vascular supply of the flap are presented. Flap healing was uncomplicated in all cases. Only 6 weeks after surgery, the flap was stable, making it possible to start adjuvant therapy and prosthetic rehabilitation swiftly. The flap is adapted to the near cone-shape of the orbit. The mean (± standard deviation) surface area of the measured orbits is (39.58 ± 3.32) cm2. The territory of the angular artery provides the periosteal flap arterial blood supply. Venous drainage is via venous networks surrounding the artery. Use of the pericranial flap makes it possible to close the orbital cavity promptly with minimal donor site defect and a short operating time, thereby minimizing the surgical risk and speeding up physical and psychological recovery.


DERIVATION OF LOCAL DIAGNOSTIC REFERENCE LEVELS FOR COMMON ADULT COMPUTED TOMOGRAPHY EXAMINATIONS IN MOROCCAN HOSPITAL.
The purpose was to derive local diagnostic reference levels (LDRLs) for adults common computed tomography (CT) examinations in Moroccan hospital. The data of patients were collected retrospectively from one facility for 2 months period. The proposed LDRLs were defined as 75th percentile of spreads for CT dose index-volume (CTDIvol) and dose length product (DLP). Findings were compared with diagnostic reference levels (DRLs) of France, Australia and Egypt. A total of 259 cases were enrolled including 53% of male and 47% of female. The corresponding LDRLs in terms of CTDIvol (mGy) were 58, 15, 15 and 14 for head, chest, abdomen-pelvis and chest-abdomen-pelvis CT examinations, respectively. For DLP (mGy.cm) were 1298, 944, 1874 and 2026, respectively. The proposed LDRLs for CTDIvol were almost similar to DRLs of Australia, higher than those of France and lower than Egyptian DRLs. In terms of DLP, LDRLs were higher than compared studies with exception of head CT, which were lower and higher for CTDIvol than Egyptian DRLs. The results highlight higher radiation doses during adult CT scans, indicating the need to spread the same investigation regarding other imaging procedures including different radiological examinations across Moroccan hospitals.


Maximal Voluntary Ventilation Should Not Be Estimated From the Forced Expiratory Volume in the First Second in Healthy People and COPD Patients.
To evaluate the concordance between the value of the actual maximum voluntary ventilation (MVV) and the estimated value by multiplying the forced expiratory volume in the first second (FEV1) and a different value established in the literature. A retrospective study was conducted with healthy subjects and patients with stable chronic obstructive pulmonary disease (COPD). Five prediction formulas MVV were used for the comparison with the MVV values. Agreement between MVV measured and MVV obtained from five prediction equations were studied. FEV1 values were used to estimate MVV. Correlation and agreement analysis of the values was performed in two groups using the Pearson test and the Bland-Altman method; these groups were one group with 207 healthy subjects and the second group with 83 patients diagnosed with COPD, respectively. We recruited 207 healthy subjects (105 women, age 47 ± 17 years) and 83 COPD patients (age 66 ± 6 years; 29 GOLD II, 30 GOLD III, and 24 GOLD IV) for the study. All prediction equations presented a significant correlation with the MVV value (from 0.38 to 0.86, p < 0.05) except for the GOLD II subgroup, which had a poor agreement with measured MVV. In healthy subjects, the mean difference of the value of bias (and limits of agreement) varied between -3.9% (-32.8 to 24.9%), and 27% (-1.4 to 55.3%). In COPD patients, the mean difference of value of bias (and limits of agreement) varied between -4.4% (-49.4 to 40.6%), and 26.3% (-18.3 to 70.9%). The results were similar in the subgroup analysis. The equations to estimate the value of MVV present a good degree of correlation with the real value of MVV, but they also show a poor concordance. For this reason, we should not use the estimated results as a replacement for the real value of MVV.


Exercise and sleep: a systematic review of previous meta-analyses.
Conduct a systematic review of previous meta-analyses on exercise and sleep outcomes in adults and a meta-analysis of studies nested within these meta-analyses. Meta-analyses of randomized controlled exercise interventions were included by searching nine electronic databases and cross-referencing. Dual-selection and data abstraction were conducted. Methodological quality of meta-analyses was assessed using AMSTAR and quality of evidence using GRADE. Random-effects models were used to pool results from the individual studies included in each meta-analysis. Three meta-analyses representing 950 adults were included. Methodological quality ranged from 36% to 64% while quality of evidence was very low to low. Statistically significant improvements (P ≤ 0.05) were observed for the apnea-hypopnea index (AHI), overall sleep quality, global score, subjective sleep, and sleep latency. The number-needed-to-treat (NNT) and percentile improvements ranged from 4 to 7 and from 18.1 to 26.5, respectively. When overall sleep quality results from individual studies nested within different meta-analyses were pooled, statistically significant standardized mean difference (SMD) improvements were observed (-0.50, 95% CI -0.72 to -0.28). The NNT and percentile improvement were 7 and 19, respectively. Exercise improves selected sleep outcomes in adults. To increase public health reach, a large, well-designed, and more inclusive meta-analysis is needed.


Antibody incubation at 37°C improves fluorescent immunolabeling in free-floating thick tissue sections.
Fluorescent immunolabeling and imaging in free-floating thick (50-60 μm) tissue sections is relatively simple in practice and enables design-based non-biased stereology, or 3-D reconstruction and analysis. This method is widely used for 3-D in situ quantitative biology in many areas of biological research. However, the labeling quality and efficiency of standard protocols for fluorescent immunolabeling of these tissue sections are not always satisfactory. Here, we systematically evaluate the effects of raising the conventional antibody incubation temperatures (4°C or 21°C) to mammalian body temperature (37°C) in these protocols. Our modification significantly enhances the quality (labeling sensitivity, specificity, and homogeneity) and efficiency (antibody concentration and antibody incubation duration) of fluorescent immunolabeling of free-floating thick tissue sections.


Obliteration of the gallbladder without formal cholecystectomy. A feasibility study.
Obliteration of the gallbladder without formal cholecystectomy was attempted in 36 rabbits, using an open technique for cannulation of the gallbladder during laparotomy. The gallbladder was cannulated, the proximal cystic duct was occluded with cyanoacrylate-nitrocellulose 2% wt/wt, and the gallbladder mucosa was exposed to various sclerosing agents to induce fibrosis. Heated 60% diatrizoate meglumine and absolute alcohol induced a controllable chronic cholecystitis with transmural fibrosis within two weeks, a result not seen with morrhuate sodium or in the control groups. All animals survived without complications, and no instances of common bile duct damage or occlusion was noted. The results reported are promising and indicate the need for further studies of this technique for its efficacy and safety.


Large-scale metabolic interaction network of the mouse and human gut microbiota.
The role of our gut microbiota in health and disease is largely attributed to the collective metabolic activities of the inhabitant microbes. A system-level framework of the microbial community structure, mediated through metabolite transport, would provide important insights into the complex microbe-microbe and host-microbe chemical interactions. This framework, if adaptable to both mouse and human systems, would be useful for mechanistic interpretations of the vast amounts of experimental data from gut microbiomes in murine animal models, whether humanized or not. Here, we constructed a literature-curated, interspecies network of the mammalian gut microbiota for mouse and human hosts, called NJC19. This network is an extensive data resource, encompassing 838 microbial species (766 bacteria, 53 archaea, and 19 eukaryotes) and 6 host cell types, interacting through 8,224 small-molecule transport and macromolecule degradation events. Moreover, we compiled 912 negative associations between organisms and metabolic compounds that are not transportable or degradable by those organisms. Our network may facilitate experimental and computational endeavors for the mechanistic investigations of host-associated microbial communities.


Increased incidence of emergency airway management after combined anterior-posterior cervical spine surgery.
Among some kinds of cervical spine surgeries, combined anterior-posterior cervical spine surgery (CAP-CS surgery) requires prolonged operative time and highly invasive procedure. This study was performed to determine whether CAP-CS surgery was associated with increased risk of emergency airway management compared with other cervical spine surgeries (O-CS surgeries). The records of the patients who underwent cervical spine surgery between July 2001 and March 2003 at our institution were reviewed retrospectively, and we determined whether the CAP-CS surgery was associated with an increased risk of emergency airway management in comparison with O-CS surgeries, using the logistic regression analysis. A total of 165 were eligible for inclusion in the study. A total of 127, 20, 11, 5, and 2 patients suffered from cervical myelopathy, traumatic cervical spinal cord injury, atlantoaxial dislocation, cervical spinal tumors, and cervical pyogenic spondylitis, respectively. The operative approaches were CAP-CS surgery, anterior surgery, posterior surgery, and atlantoaxial surgery in 10, 56, 88, and 11 patients, respectively. Thus, the operative approaches were CAP-CS surgery in 10 patients and O-CS surgeries in 155 patients. Postoperative emergency airway management was required in 7 of the 10 patients (70%) who underwent CAP-CS surgery, and 2 of the 155 patients (1%) who underwent O-CS surgeries. The increased risk of postoperative emergency airway management imposed by CAP-CS surgery was 178.5 by an odds ratio, with a 95% confidence interval of 25.6 to 1246. The results show that CAP-CS surgery provides a major risk factor for postoperative emergency airway management.


T cell receptor junctional regions and the MHC molecule affect the recognition of antigenic peptides by T cell clones.
Nine independent pigeon cytochrome c-specific T cell clones were analyzed by using a panel of antigenic peptide analogs presented in association with three allelic IE-encoded MHC glycoproteins. Eight of the T cell clones expressed a TCR composed of a unique alpha- and beta-chain amino acid sequence, and concordantly, each of these T cell clones exhibited a unique Ag specificity. This was true for several clones which differed only in TCR V-J junctional regions. Interestingly, for a given clone, the response to some of the peptide analogs depended to a large extent on the allelic form of the presenting MHC molecule. A simple interpretation of these data would suggest that certain positions of the peptide Ag are most important for Ag-MHC molecule interactions, and that these specific interactions can influence the antigenic epitope recognized by the TCR. We suggest that an antigenic peptide binds to an MHC glycoprotein in a distinct way, but may retain a measure of flexibility.


Halogen-Adjusted Chemoselective Synthesis of Fluorene Derivatives with Position-Controlled Substituents.
Fluorenes have been synthesized through an efficient novel Pd-catalyzed tandem cross-coupling reaction; these substrates are fascinating building blocks found in organic photoelectric materials. The position of the substituent on fluorenes could be conveniently tuned by changing the halogen in the ortho-halobenzyl bromide substrates when coupled with various arylboronic acids. This newly developed synthetic approach could achieve the potential diversity in fluorene-based molecular architectures.


Potential role of QT interval prolongation in sudden infant death syndrome.
To investigate the possibility that a genetically transmitted cardiac abnormality is involved in the genesis of the sudden infant death syndrome (SIDS), 42 sets of parents who had at least one infant with SIDS were studied by electrocardiography. Prolongation of the QT interval was present in at least one member of 11 (26%) sets of parents. In families in which QT interval prolongation was found in a parent, prolonged QT interval was also present in 39% of the siblings of infants with SIDS, suggesting an autosomal dominant pattern of inheritance. In addition, an infant with "near-miss" SIDS showed marked prolongation of the QT interval. Thus, our data suggest that prolonged QT interval may play a role in a considerable proportion of sudden and unexpected infant deaths. However, definitive confirmation of the relation between QT interval prolongation and SIDS will require large prospective investigations.


Using Electronic Health Records to Build an Ophthalmologic Data Warehouse and Visualize Patients' Data.
To develop a near-real-time data warehouse (DW) in an academic ophthalmologic center to gain scientific use of increasing digital data from electronic medical records (EMR) and diagnostic devices. Database development. Specific macular clinic user interfaces within the institutional hospital information system were created. Orders for imaging modalities were sent by an EMR-linked picture-archiving and communications system to the respective devices. All data of 325 767 patients since 2002 were gathered in a DW running on an SQL database. A data discovery tool was developed. An exemplary search for patients with age-related macular degeneration, performed cataract surgery, and at least 10 intravitreal (excluding bevacizumab) injections was conducted. Data related to those patients (3 142 204 diagnoses [including diagnoses from other fields of medicine], 720 721 procedures [eg, surgery], and 45 416 intravitreal injections) were stored, including 81 274 optical coherence tomography measurements. A web-based browsing tool was successfully developed for data visualization and filtering data by several linked criteria, for example, minimum number of intravitreal injections of a specific drug and visual acuity interval. The exemplary search identified 450 patients with 516 eyes meeting all criteria. A DW was successfully implemented in an ophthalmologic academic environment to support and facilitate research by using increasing EMR and measurement data. The identification of eligible patients for studies was simplified. In future, software for decision support can be developed based on the DW and its structured data. The improved classification of diseases and semiautomatic validation of data via machine learning are warranted.


Posterior Cortical Atrophy Presenting with Superior Arcuate Field Defect.
An 80-year-old female with reading difficulty presented with progressive arcuate field defect despite low intraocular pressure. Over a 5-year period, the field defect evolved into an incongruous homonymous hemianopia and the repeated neuroimaging revealed progressive posterior cortical atrophy. Further neuropsychiatric assessment demonstrated symptoms and signs consistent with Benson's syndrome.


Leukemia in severe congenital neutropenia: defective proteolysis suggests new pathways to malignancy and opportunities for therapy.
Severe congenital neutropenia (SCN), a heterogeneous disorder that includes Kostmann syndrome, predisposes to myelodysplasia and acute myelogenous leukemia. Recently identified heterozygous mutations in the gene ELA2, encoding neutrophil elastase on human chromosome 19pter, account for the majority of autosomal dominant cases of SCN, including those demonstrating neoplastic progression. The involvement of the serine protease neutrophil elastase, localized to the granules of neutrophils and monocytes, implies an unexpected role for proteolytic regulation of hematopoiesis. Continued elucidation of the clinical features, molecular genetics, and biochemistry is likely to provide insight into novel pathways of leukemia induction with attendant prospects for new avenues of therapy.


Effects of testosterone therapy on BMI, blood pressure, and laboratory profile of transgender men: a systematic review.
Testosterone is the main hormonal agent used for cross-sex hormone therapy in female-to-male transgender persons. Our aim was to systematically review the literature concerning the effects of testosterone on body mass index (BMI), blood pressure, hematocrit, hemoglobin, lipid profile, and liver enzymes in transgender men. PUBMED and EMBASE were searched for studies published until March 2017. Studies were included if they reported interventions with any dose of testosterone and comparison of variables before and during treatment. Of 455 potentially eligible articles, 13 were reviewed. Study duration ranged from 6 to 60 months, sample size ranged from 12 to 97 patients, and the most common treatment was parenteral testosterone undecanoate 1000 mg/12 weeks. Slight but significant increases in BMI were reported (from 1.3 to 11.4%). Three out of seven studies assessing the impact of different testosterone formulations on blood pressure detected modest increases or clinically irrelevant changes in this variable. In another study, however, two patients developed hypertension, which was resolved after cessation of testosterone therapy. Decreases in HDL-cholesterol and increases in LDL-cholesterol were consistently observed. Eight studies observed a relationship between testosterone and increased hemoglobin (range: 4.9-12.5%) and hematocrit (range: 4.4-17.6%), but discontinuation of androgen therapy was not necessary. In one study, two patients developed erythrocytosis (hematocrit >52%) after 9 and 12 months of treatment. One study analyzing testosterone formulations observed smaller increases in hemoglobin and hematocrit with testosterone gel. Six studies assessing liver function showed slight or no changes. Overall, the quality of evidence was low, given the lack of randomized clinical/controlled trials and the small sample sizes. In conclusion, exogenous testosterone administration to transgender men was associated with modest increases in BMI, hemoglobin/hematocrit, and LDL-cholesterol, and with decreases in HDL-cholesterol. Long-term studies are needed to assess the long-term risks of testosterone therapy, particularly as they relate to cardiometabolic risks such as diabetes, dyslipidemia and the metabolic syndrome.


Small-molecule TrkB receptor agonists improve motor function and extend survival in a mouse model of Huntington's disease.
Huntington's disease (HD) is a fatal neurodegenerative disease characterized by abnormal motor coordination, cognitive decline and psychiatric disorders. This disease is caused by an expanded CAG trinucleotide repeat in the gene encoding the protein huntingtin. Reduced levels of brain-derived neurotrophic factor (BDNF) in the brain, which results from transcriptional inhibition and axonal transport deficits mediated by mutant huntingtin, have been suggested as critical factors underlying selective neurodegeneration in both HD patients and HD mouse models. BDNF activates its high-affinity receptor TrkB and promotes neuronal survival; restoring BDNF signaling is thus of particular therapeutic interest. In the present study, we evaluated the ability of a small-molecule TrkB agonist 7,8-dihydroxyflavone (7,8-DHF) and its synthetic derivative 4'-dimethylamino-7,8- dihydroxyflavone (4'-DMA-7,8-DHF) to protect neurons in the well-characterized N171-82Q HD mouse model. We found that chronic administration of 7, 8-DHF (5 mg/kg) or 4'-DMA-7,8-DHF (1 mg/kg) significantly improved motor deficits, ameliorated brain atrophy and extended survival in these N171-82Q HD mice. Moreover, 4'-DMA-7,8-DHF preserved DARPP32 levels in the striatum and rescued mutant huntingtin-induced impairment of neurogenesis in the N171-82Q HD mice. These data highlight consideration of TrkB as a therapeutic target in HD and suggest that small-molecule TrkB agonists that penetrate the brain have high potential to be further tested in clinical trials of HD.


Ethical Issues in Commercialisation of Stem Cell Therapies: Mapping the Terrain.
Bioethical debates on stem cell research have focused primarily on the moral status of human embryos. This article highlights seven distinct policy and ethical issues associated with the commercialisation of stem cell therapies, describes some of the underlying moral questions on which they turn, and argues that there is an urgent need to refocus the debate on stem cell research beyond the controversy over embryo destruction.


Calmodulin expression during rat liver regeneration.
We have investigated the messenger RNAs expressed from the three calmodulin genes during rat liver regeneration. The results revealed that all the calmodulin transcripts increased from 8 hr after a partial hepatectomy, although differences in the timing and the level of expression from the three genes were observed. Calmodulin I transcripts peaked at 16 hr, whereas calmodulin II and calmodulin III progressively increased from 8 to 24 hr. At 24 hr after surgery, calmodulin I, calmodulin II and the 2.3 kb calmodulin III transcripts reached values of a 6-fold increase, whereas the 0.8 kb product of calmodulin III increased 25-fold. At 30 hr the levels of all the calmodulin transcripts were similar to those observed at 24 hr. The transcription rates of the three calmodulin genes augmented after hepatectomy (calmodulin I and calmodulin II twofold and calmodulin III fourfold), indicating that the elevation of the calmodulin transcripts could be, at least partially, the result of this increase in the transcription rates. The total calmodulin concentration also increased twofold at 24 hr after hepatectomy. We also report that the administration of the beta-adrenergic blocker, D,L-propranolol inhibited the accumulation of calmodulin protein without significantly affecting the increase of the messenger RNAs. These results indicate that the expression of calmodulin observed during liver regeneration could be regulated by cyclic AMP at the translational or posttranslational level.


Hydrogen Sulfide Inhibits Autophagic Neuronal Cell Death by Reducing Oxidative Stress in Spinal Cord Ischemia Reperfusion Injury.
Autophagy is upregulated in spinal cord ischemia reperfusion (SCIR) injury; however, its expression mechanism is largely unknown; moreover, whether autophagy plays a neuroprotective or neurodegenerative role in SCIR injury remains controversial. To explore these issues, we created an SCIR injury rat model via aortic arch occlusion. Compared with normal controls, autophagic cell death was upregulated in neurons after SCIR injury. We found that autophagy promoted neuronal cell death during SCIR, shown by a significant number of terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling- (TUNEL-) positive cells colabeled with the autophagy marker microtubule-associated protein 1 light chain 3, while the autophagy inhibitor 3-methyladenine reduced the number of TUNEL-positive cells and restored neurological and motor function. Additionally, we showed that oxidative stress was the main trigger of autophagic neuronal cell death after SCIR injury and N-acetylcysteine inhibited autophagic cell death and restored neurological and motor function in SCIR injury. Finally, we found that hydrogen sulfide (H2S) inhibited autophagic cell death significantly by reducing oxidative stress in SCIR injury via the AKT-the mammalian target of rapamycin (mTOR) pathway. These findings reveal that oxidative stress induces autophagic cell death and that H2S plays a neuroprotective role by reducing oxidative stress in SCIR.


Tails of the crossing probability.
The scaling of the tails of the probability of a system to percolate only in the horizontal direction pi(hs) was investigated numerically for the correlated site-bond percolation model (q -state Potts model) for q=1 , 2, 3, 4 (where q is the number of spin states). We have to demonstrate that the crossing probability pi(hs) (p) far from the critical point p(c) has the shape pi(hs) (p) similar to D exp [cL (p- p(c) )(nu) ] where nu is the correlation length index, and p=1-exp (-beta) is the probability of a bond to be closed. For the tail region the correlation length is smaller than the lattice size. At criticality the correlation length reaches the sample size and we observe crossover to another scaling pi(hs) (p) similar to A exp {-b [L (p- p(c) )(nu)](x)}. Here x is a scaling index describing the central part of the crossing probability.


Hampton, Nutting, and Rival gospels at The Johns Hopkins Hospital and Training School for Nurses, 1889-1906.
To examine the attempts by Isabel Hampton Robb and Mary Adelaide Nutting to reshape nurse training into an academic system of nursing education. As health care delivery undergoes transformation in the 1990s, it is useful to consider the strategies used as well as the successes and failures, of past nurse leaders. Historical research. Archival data were analyzed in 1996 pertaining to the creation in 1889 and administration of the nursing school and hospital founded by Johns Hopkins in Baltimore, Maryland, USA. Events were studied using published and unpublished writings by Hampton and Nutting, as well as relevant secondary sources. Separated by class from those with power, and limited by 19th century notions of women's place, Isabel Hampton Robb and Adelaide Nutting helped inaugurate nursing's quest for professionalization. Undaunted by adversity, they achieved novel accomplishments for women and nurses. Hampton and Nutting possessed vision, intelligence, the capacity to motivate, political skills, good judgement, and managerial competence. Hampton's and Nutting's inability to strengthen the link between the school of nursing and the Johns Hopkins University and the lack of unity within nursing limited their success. This research illuminates some of the controversies pertaining to nursing during an earlier period of sweeping health care change.


Nurse-led pre-operative assessment for elective surgical patients.
Effective pre-operative assessment improves patient outcomes by ensuring that individuals are adequately prepared for anaesthesia, surgery and the post-operative period. It can also improve hospital efficiency by reducing patient waiting times and enabling early discharge. Nurse-led pre-operative assessment for elective surgery is becoming increasingly recognised as a valuable method of ensuring that patients have a safe and well-planned hospital stay. This article examines the role of the pre-operative assessment nurse within a nurse-led service for elective surgery at a district general hospital.


Increase of basic fibroblast growth factor (FGF) and FGF receptor messenger RNA during rat thyroid hyperplasia: temporal changes and cellular distribution.
Goitre was induced in adult rats by acute (1 or 2 weeks) or chronic (4 or 10 weeks) administration of methimazole together with a low iodine diet. Involution of thyroid growth was then observed at 16 weeks, 4 weeks after withdrawal of goitrogens and reversion to a normal diet. Experimental animals quickly became hypothyroid compared with controls and exhibited thyroid hyperplasia (control (n = 10): total serum thyroxine (T4) 66 +/- 4 nmol/l, thyroid weight 5 +/- 1 mg/100 g body weight, means +/- S.D.; experimental (n = 10): T4 undetectable, thyroid weight 27 +/- 4 mg/100 g body weight after 2 weeks of treatment). Thyroid growth rate subsequently slowed between 2 and 10 weeks. Messenger RNA for basic fibroblast growth factor (basic FGF) and for the high-affinity FGF receptor, was compared in the thyroids and livers of control and goitrous rats by ribonuclease protection assay. Low levels of mRNA for basic FGF and its receptor were detectable in thyroids from control rats at all times, while none was detected in the livers from any animal. Basic FGF and receptor mRNAs increased, and were detected at greatest abundance in hyperplastic thyroids at 1 and 2 weeks respectively, during goitre formation, but subsequently declined in parallel with thyroid growth rate at 4 and 10 weeks. When quantified by radioimmunoassay, basic FGF extracted from thyroids was fivefold greater than in controls after 1 week of goitrogen treatment (control (n = 4): 24 +/- 9 pmol/micrograms DNA; goitre (n = 4): 100 +/- 16 pmol/micrograms DNA; P < 0.05). Basic FGF and FGF receptor mRNAs localized by in situ hybridization predominantly to the epithelial cell population within follicles. Localization by immunohistochemistry demonstrated that basic FGF was present in the thyroids of control rats, and was largely associated with the basement membrane of follicles. During thyroid hyperplasia, increased basic FGF immunoreactivity appeared over the cytoplasm of follicular epithelial cells and was lost from the extracellular matrix. Thyroid involution following removal of goitrogen/low iodine treatment was associated with a decrease in mRNA for basic FGF or its receptor, and a loss of immunoreactive basic FGF from the cytoplasm of follicular cells. These results suggest that autocrine expression of basic FGF and FGF receptor could contribute to thyroid hyperplasia in rats.


Temperature effects on ion association and hydration in MgSO4 by dielectric spectroscopy.
A detailed investigation of aqueous solutions of magnesium sulfate has been made by dielectric relaxation spectroscopy (DRS) over a wide range of frequencies (0.2<or=nu/GHz<or=89), concentrations (0.02<or=m/mol kg(-1)<or=2.3) and temperatures (5<or=theta/ degrees C<or=65). Consistent with literature data from other techniques, the spectra show clear evidence at all temperatures for the simultaneous existence of double solvent-separated (2SIP), solvent-shared (SIP) and contact (CIP) ion pairs and a triple ion (TI), Mg(2)SO(4) (2+)(aq). Increased formation of CIPs and especially TIs occurs with increasing temperature. The overall standard ion-association constant, K(A) degrees , corresponding to the equilibrium: Mg(2+)(aq)+SO(4) (2) (-)(aq) <==> MgSO(4) (0)(aq) is in good agreement with literature data at lower temperatures but is overestimated at higher temperatures due to processing difficulties. Despite the limited precision of the spectra, analysis of the individual steps in the ion-association process is possible for the first time. The 2SIPs are formed with little disturbance to their hydration shells, the (partial) destruction of which appears to occur mostly during the formation of SIPs. Effective hydration numbers derived from the DRS spectra indicate that both Mg(2+) and SO(4) (2-) influence solvent water molecules beyond their first hydration spheres but that MgSO(4)(aq) is less strongly hydrated than the previously studied CuSO(4)(aq).


The results of treatment of epithelial ovarian cancer after centralisation of primary surgery. Results from North Jutland, Denmark.
The study was performed to evaluate the results of treatment of ovarian carcinoma after the introduction of centralised primary surgery in the County of North Jutland, Denmark. Prospective study of consecutive cases of ovarian cancer undergoing primary surgical treatment at the Gynecologic Oncologic Center after the introduction of centralised primary surgery. Results of treatment recorded up to the date of last examination or death. From 1999 to 2002, 107 patients with primary epithelial ovarian cancer underwent primary surgery at the Gynecologic Oncologic Center, Aalborg. This corresponds to 95.5% of patients with invasive carcinoma in the County of North Jutland. All patients with Stage I to Stage IIIB disease had a complete, macroscopically radical cytoreduction performed. In patients with Stage III and IV invasive tumors, the optimal debulking rate was 79.5%, and, in Stage IIIC and IV, the optimal debulking rate was 78.2%. Intra-operative and post-operative complications were generally few. Post-operative death, defined as death within 30 days after surgery, was observed in 4 cases (3.7%). After primary surgery, platinum-based chemotherapy was given in most cases. For Stage I to IV invasive cancer, the median survival was 46 months. In patients with Stage IIIC and IV disease, the median survival was 32 months. In optimally debulked Stage IIIC and IV disease, the median survival was 41 months. The results indicate a survival benefit after introduction of centralised primary surgery. Compared to existing national and regional data on survival in ovarian cancer, the results indicate an increase in median survival for all stages of approximately 15 months. Centralisation of primary surgery to centres with the necessary expertise may be the most significant way to increase survival in ovarian cancer in Denmark.


Rotator cuff-related pain: Patients' understanding and experiences.
Persistent musculoskeletal pain is a multi-factorial entity, influenced by biological, genetic and psychosocial factors. Psychosocial factors, such as individuals' beliefs and experiences, need to be considered in the management of such pain. While extensive research has explored beliefs of individuals with spinal pain, less is known about individuals' beliefs regarding shoulder pain. To explore beliefs about the cause of pain in individuals with persistent rotator cuff-related pain, as well as the experiences of the effect of pain on their daily lives. A mixed methods design, using semi-structured interviews and validated outcome questionnaires. Five men and five women, aged 47-68 years, with shoulder pain for at least three months were recruited. Individual semi-structured interviews were audio-recorded, transcribed verbatim and analysed using the general inductive approach. Four key themes emerged. The cause of pain, 'Understanding the pain', was described in terms of anatomical factors within the context of the participants' lives. The pain impacted all areas of life, creating another theme, 'It affects everything'. Participants responded to their pain by adopting certain, 'Pain-associated behaviours' and sought information for diagnosis, general management and exercise prescription, 'Emotional responses and the future'. The participants with rotator cuff-related pain believed the cause of their pain to be local to the shoulder region. However, they also described various stressors in their work-, sports- and family-related lives. Rehabilitation may need to include educating the individual, expanding their understanding regarding pain mechanisms and appropriate interventions, based on individual goal-setting.


Biomarker for Spinal Muscular Atrophy: Expression of SMN in Peripheral Blood of SMA Patients and Healthy Controls.
Spinal muscular atrophy is caused by a functional deletion of SMN1 on Chromosome 5, which leads to a progressive loss of motor function in affected patients. SMA patients have at least one copy of a similar gene, SMN2, which produces functional SMN protein, although in reduced quantities. The severity of SMA is variable, partially due to differences in SMN2 copy numbers. Here, we report the results of a biomarker study characterizing SMA patients of varying disease severity. SMN copy number, mRNA and Protein levels in whole blood of patients were measured and compared against a cohort of healthy controls. The results show differential regulation of expression of SMN2 in peripheral blood between patients and healthy subjects.


Randomized clinical trial comparing hospital to ambulatory rehabilitation of malnourished children in Niger.
This study compared hospital to ambulatory nutritional rehabilitation outcomes and costs. Following a hospital stay to resolve initial acute medical conditions, 100 malnourished children (54 per cent male, ages 5 to 28 months) in Niger were randomly assigned to either hospital or ambulatory nutritional rehabilitation. Anthropometric measures were assessed at 15, 30, 60, 90 and 180 days post-randomization. Following randomization, the hospital group received a mean of 12.9 days of hospital rehabilitation and 5.6 days of ambulatory rehabilitation, while the ambulatory group received 2.2 days of hospital rehabilitation and 11.9 days of ambulatory rehabilitation. No significant differences between the two study groups in mortality rates or weight gain were found. The mean cost for hospital rehabilitation was 120 per cent higher (P < 0.001) than ambulatory rehabilitation. This study was the first randomized clinical trial directly comparing hospital to ambulatory nutritional rehabilitation and suggests that ambulatory rehabilitation is more cost-effective.


[New aspects of liver scintigraphy (author's transl)].
The development of the scintillation camera computer scintigraphy has created new chances for nuclear medicine diagnosis of the liver. With the development of new 99mTc-labeled radiopharmacenticals the effectiveness of functional scintigraphy of the liver in the differential diagnosis of parenchymal and mechanical icterus could be considerably increased. Computer perfusion scintigraphy permits proof of circulatory changes in the liver without invasive techniques. Its importance lies in early diagnosis of portal hypertension and in pre- and post-operative assessment of liver circulation after shunt surgery.


Upregulated miR-206 Aggravates Deep Vein Thrombosis by Regulating GJA1-Mediated Autophagy of Endothelial Progenitor Cells.
Deep vein thrombosis (DVT) is the third most prevalent vascular disease worldwide. MicroRNAs (miRNAs) play regulatory roles in functions of endothelial progenitor cells (EPCs), which is becoming a promising therapeutic choice for thrombus resolution. Nevertheless, the role of miR-206 in EPCs is unclear. EPCs were isolated from the peripheral blood of patients with DVT. In DVT mouse models, DVT was induced by stenosis of the inferior vena cava (IVC). The levels of miR-206 and gap junction protein alpha 1 (GJA1) in EPCs and vascular tissues of DVT mice were detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The proliferation, migration, apoptosis, and angiogenesis were tested by cell counting kit-8 (CCK-8) assay, Transwell assay, flow cytometry analysis, and in vitro tube formation assay. The levels of autophagy-related proteins as well as the level of GJA1 in EPCs and vascular tissues were evaluated by western blotting. DVT formation in vivo was observed through hematoxylin-eosin (HE) staining. The expression of thrombus resolution markers, CD34 molecule (CD34) and matrix metallopeptidase 2 (MMP2), in the thrombi was measured by immunofluorescence staining. miR-206 overexpression inhibited proliferation, migration, and angiogenesis and promoted apoptosis of EPCs, while miR-206 knockdown exerted an opposite effect on EPC phenotypes. Downregulation of GJA1, the target of miR-206, abolished the influence of miR-206 on EPC phenotypes. Furthermore, silencing of miR-206 suppressed the autophagy of EPCs via upregulating GJA1. miR-206 knockdown repressed thrombus formation, enhanced the homing ability of EPCs to the thrombosis site, and facilitated thrombus resolution in DVT mouse models. Additionally, miR-206 was upregulated while GJA1 was downregulated in vascular tissues of DVT mice. miR-206 knockdown elevated GJA1 expression in vascular tissues of DVT mice. The expression of miR-206 was negatively correlated with that of GJA1 in DVT mice. miR-206 knockdown upregulates GJA1 to inhibit autophagy of EPCs and then promote EPC proliferation, migration, and angiogenesis, thereby enhancing EPC homing to thrombi and facilitating thrombus resolution.


Collagen synthesis is upregulated in mitral valves subjected to altered stress.
Mitral regurgitation (MR) and abnormal ventricular wall motion (AVWM) are two cardiac conditions that may increase mitral valve (MV) stresses. Theoretically, increased stress could induce damaging MV tissue alterations. These alterations may impair the preferred option of repair, and mandate replacement. It is hypothesized that MV collagen synthesis is upregulated in response to MR and AVWM. To test this hypothesis in a pilot study, an ischemic sheep model (n = 8) was employed. Four sheep underwent selective coronary artery ligation to infarct a papillary muscle, which resulted in MR. Two other sheep underwent similar coronary ligation to create AVWM. As controls, two sheep underwent sham surgery (no ligation). Sheep were killed 4 and 8 weeks post operatively and their MVs were sectioned. Sections were stained with an antibody (SP1.D8, University of Iowa) to procollagen I (precursor to collagen I). The percent area of procollagen stain present present was measured by image analysis (Optimas Corporation) and used as an indicator of collagen synthesis. Procollagen results indicated that MV collagen synthesis was upregulated by factor of 1.8 in both the MR and AVWM groups versus controls. In addition, results showed greater upregulation in anterior leaflets compared with posterior leaflets in both infarct groups. These results indicate that MV collagen synthesis is upregulated in response to MR and AVWM.


A retrotransposon insertion in the 5' regulatory domain of Ptf1a results in ectopic gene expression and multiple congenital defects in Danforth's short tail mouse.
Danforth's short tail mutant (Sd) mouse, first described in 1930, is a classic spontaneous mutant exhibiting defects of the axial skeleton, hindgut, and urogenital system. We used meiotic mapping in 1,497 segregants to localize the mutation to a 42.8-kb intergenic segment on chromosome 2. Resequencing of this region identified an 8.5-kb early retrotransposon (ETn) insertion within the highly conserved regulatory sequences upstream of Pancreas Specific Transcription Factor, 1a (Ptf1a). This mutation resulted in up to tenfold increased expression of Ptf1a as compared to wild-type embryos at E9.5 but no detectable changes in the expression levels of other neighboring genes. At E9.5, Sd mutants exhibit ectopic Ptf1a expression in embryonic progenitors of every organ that will manifest a developmental defect: the notochord, the hindgut, and the mesonephric ducts. Moreover, at E 8.5, Sd mutant mice exhibit ectopic Ptf1a expression in the lateral plate mesoderm, tail bud mesenchyme, and in the notochord, preceding the onset of visible defects such as notochord degeneration. The Sd heterozygote phenotype was not ameliorated by Ptf1a haploinsufficiency, further suggesting that the developmental defects result from ectopic expression of Ptf1a. These data identify disruption of the spatio-temporal pattern of Ptf1a expression as the unifying mechanism underlying the multiple congenital defects in Danforth's short tail mouse. This striking example of an enhancer mutation resulting in profound developmental defects suggests that disruption of conserved regulatory elements may also contribute to human malformation syndromes.


Disability caused by work-related musculoskeletal disorders.
This article describes the magnitude, extent, and economic consequences of some of the more common, work-related musculoskeletal disorders. In addition, it provides a brief historic overview of the state-federal vocational rehabilitation program in the United States. It identifies and considers a constellation of risk factors for work-related disability because of musculoskeletal disorders, and it discusses phases of physical rehabilitation as that process relates to injured workers. The shifting disability paradigm is examined, and attention is given to terminology that has become fashionable since passage of the landmark Americans with Disabilities Act of 1990. Finally, various factors and conditions that often become barriers to an injured person's successful return to the workforce are briefly discussed.


[Biochemical study on anti-inflammatory action of anti-allergic drugs--with regard to NADPH oxidase].
The effects of anti-allergic drugs with or without H1-receptor antagonism on the NADPH oxidase from human neutrophils in both whole-cell and fully soluble (cell-free) systems were investigated. Three anti-allergic drugs with H1-receptor antagonism, azelastine, ketotifen and oxatomide, were found to inhibit the superoxide generation of human neutrophils exposed to phorbol myristate acetate in a whole-cell system and the activation of superoxide-generating NADPH oxidase by sodium dodecyl sulfate in a cell-free system. The concentrations of three drugs required for 50% inhibition of the oxidase (IC50) were as follows: azelastin--0.7 microM in the whole-cell system and 0.5 microM in the cell-free system, ketotifen--60 microM in the whole-cell system and 6.8 microM in the cell-free system, and oxatomide--25 microM in the whole-cell system and 9.7 microM in the cell-free system. In addition, in the cell-free system, these drugs did not change the Km values for the NADPH of the oxidase. However, micromoles of tranilast, an anti-allergic drug without H1-receptor antagonism, did not inhibit neutrophil NADPH oxidase in the whole-cell and cell-free systems. The IC50 of hydrocortisone in the cell-free system was 60 microM. These results suggest that anti-allergic drugs with H1-receptor antagonism inhibit reconstitution of the solubilized membrane-bound enzyme by sodium dodecyl sulfate in cell-free systems and that they have a strong anti-inflammatory action. Anti-allergic drugs are not the drugs of first choice for asthma, but these drugs, especially basic anti-allergic drugs, should be used more frequently for the treatment of chronic asthma, infectious-typed asthma and mixed-typed asthma closely associated with acute and chronic inflammation of the airways as well as atopic asthma.


Sporulation and time course expression of sigma-factor homologous genes in Clostridium acetobutylicum.
The gene for the vegetative sigma factor A of Clostridium acetobutylicum was constitutively transcribed during growth and formed an operon together with dnaE. Sporulation-specific sigma factors E, G, and K were sequentially induced shortly before mature endospores could be detected. Maximal transcription in the course of spore formation was found to be in the order sigE-sigG-sigK, thus matching the pattern described for Bacillus subtilis. From primer extension experiments promoter structures could be deduced with high homology to the Bacillus consensus. Upstream of the spoIIGA-sigE operon a gene with significant similarity to ftsZ could be detected.


Upregulation of Sortilin, a Lysosomal Sorting Receptor, Corresponds with Reduced Bioavailability of Latent TGFβ in Mucolipidosis II Cells.
Mucolipidosis II (ML-II) is a lysosomal disease caused by defects in the carbohydrate-dependent sorting of soluble hydrolases to lysosomes. Altered growth factor signaling has been identified as a contributor to the phenotypes associated with ML-II and other lysosomal disorders but an understanding of how these signaling pathways are affected is still emerging. Here, we investigated transforming growth factor beta 1 (TGFβ1) signaling in the context of ML-II patient fibroblasts, observing decreased TGFβ1 signaling that was accompanied by impaired TGFβ1-dependent wound closure. We found increased intracellular latent TGFβ1 complexes, caused by reduced secretion and stable localization in detergent-resistant lysosomes. Sortilin, a sorting receptor for hydrolases and TGFβ-related cytokines, was upregulated in ML-II fibroblasts as well as GNPTAB-null HeLa cells, suggesting a mechanism for inappropriate lysosomal targeting of TGFβ. Co-expression of sortilin and TGFβ in HeLa cells resulted in reduced TGFβ1 secretion. Elevated sortilin levels correlated with normal levels of cathepsin D in ML-II cells, consistent with a compensatory role for this receptor in lysosomal hydrolase targeting. Collectively, these data support a model whereby sortilin upregulation in cells with lysosomal storage maintains hydrolase sorting but suppresses TGFβ1 secretion through increased lysosomal delivery. These findings highlight an unexpected link between impaired lysosomal sorting and altered growth factor bioavailability.


Unraveling the electron transfer processes of a nanowire protein from Geobacter sulfurreducens.
The extracellular electron transfer metabolism of Geobacter sulfurreducens is sustained by several multiheme c-type cytochromes. One of these is the dodecaheme cytochrome GSU1996 that belongs to a new sub-class of c-type cytochromes. GSU1996 is composed by four similar triheme domains (A–D). The C-terminal half of the molecule encompasses the domains C and D, which are connected by a small linker and the N-terminal half of the protein contains two domains (A and B) that form one structural unit. It was proposed that this protein works as an electrically conductive device in G. sulfurreducens, transferring electrons within the periplasm or to outer-membrane cytochromes. In this work, a novel strategy was applied to characterize in detail the thermodynamic and kinetic properties of the hexaheme fragment CD of GSU1996. This characterization revealed the electron transfer process of GSU1996 for the first time, showing that a heme at the edge of the C-terminal of the protein is thermodynamic and kinetically competent to receive electrons from physiological redox partners. This information contributes towards understanding how this new sub-class of cytochromes functions as nanowires, and also increases the current knowledge of the extracellular electron transfer mechanisms in G. sulfurreducens.


Stapedius reflex thresholds obtained in a free sound field as an indicator for over- and understimulation in cochlear implant listeners.
The goal of this study was to determine open field stapedius reflex thresholds (oSRTs) in CI patients with fittings based on subjective loudness ratings. A further objective was to compare these oSRTs and those of eSRT-based fittings that are similar to the oSRTs of normal hearing. Impedance measurements of the ear drum were taken while subjects were wearing their audio processors. The stapedius reflex was elicited by electrical stimulation transmitted through the activated CI system in response to an acoustic stimulus presented in the free sound field. Subjects were 50 experienced CI users (n = 57 ears) with CI fittings based on subjective loudness scaling. A reference range for the oSRTs was defined that was identified in CI patients with eSRT-based fittings. Sound levels for stapedius reflex detection were inside the reference target range in 70% of the cases, below the reference range (i.e. down to 40 dB HL) in 20% of the cases, and above the reference range in 10% of the cases. Stapedius reflex detection in a free sound field may help detect fittings with too high or too low stimulation levels that might reduce audiological performance.


Effects of PEMF and glucocorticoids on proliferation and differentiation of osteoblasts.
Pulsed electromagnetic fields (PEMF) can promote bone healing, while use of dexamethasone induces bone loss and osteoporosis. There is no report available on the combined effects of PEMF and dexamethasone on the activity of osteoblasts. Here, we investigated the effects of PEMF and dexamethasone on the proliferation and differentiation of MC3T3-E1 osteoblasts. Our results showed that PEMF and dexamethasone respectively increased and decreased the proliferation of MC3T3-E1 osteoblasts, meanwhile PEMF eliminated the effect of dexamethasone on MC3T3-E1 osteoblasts. Moreover, we also found that dexamethasone combined with PEMF upregulated the mRNA expression of IGF-1 at the early stage after the stimulation of PEMF and improved the decrease of COX-2 mRNA expression induced by dexamethasone at the late stage after the stimulation of PEMF. PEMF may be beneficial to improve dexamethasone-induced bone loss and osteoporosis.


Photoaffinity labeling of the ribosomal A site with S-(p-azidophenacyl)valyl-tRNA.
S-(p-Azidophenacyl)valyl-tRNA, an analog of valyl-tRNA which has a photoaffinity label attached to its 4-thiouridine residue, was bound to the ribosomal A site at 10 mM Mg2+. Binding was stimulated 25-fold by the presence of elongation factor EFTu. Photoactivation of the p-azidophenacyl group by irradiation resulted in covalent linking of 6% of the noncovalently bound tRNA to the ribosomes. Covalent linking was dependent on the simultaneous presence of ribosomes, poly(U2,G),EFTu.GTP, required irradiation, and did not occur when S-(phenacyl)valyl-tRNA, a nonphotolyzable analog, replaced S-(p-azidophenacyl)valyl-tRNA. The attached tRNA was distributed approximately equally between both the 30S and 50S subunits. At the 30S subunit, 30% of the tRNA was bound to protein while 70% was linked to 16S RNA. At the 50S subunit, however, negligible binding to the 23S RNA was observed. More than 90% of the tRNA was attached to low molecular weight material according to sodium dodecyl sulfate-sucrose gradient analysis, and more than 87% of this fraction consisted of tRNA-protein complexes as assayed by phenol solubility and electrophoretic mobility before and after protease treatment. These results, in conjunction with our previous report (I. Schwartz and J Ofengand (1974), Proc. Natl. Acad. Sci. U.S.A. 71, 3951) which showed that covalent linking of this same tRNA derivative at the ribosomal P site resulted in attachment solely to the 16S RNA, demonstrate that 16S, but not 23S or 5S rRNA, is an important component of the tRNA binding site in the region of the 4-thiouridine residue and furthermore show that ribosomal A and P sites are topologically distinct.


[Emerging Therapeutic Targets Revealed by Genome Analysis in Alzheimer's Disease].
Cutting-edge genomics technologies have substantially improved our understanding of the pathogenesis of Alzheimer's disease (AD). The identification of mutations in APP, PSEN1, and PSEN2 causative for autosomal dominant AD (ADAD) has provided the basis of the "amyloid cascade" hypothesis of the pathogenetic mechanism of AD. While a number of therapeutic candidates targeting amyloid-β were developed using genetically engineered mouse models of ADAD, none have proven successful in mitigating the symptoms of AD in phase III clinical trials. Thus, we must reconsider the modification of AD pathogenesis by targets within the amyloid cascade, and explore new targets for AD therapy. This review provides a comprehensive summary of the recent genetic studies of AD from this perspective, and discusses prospective and emerging therapeutic interventions for patients with AD.


Nucleophile-Assisted Alkene Activation: Olefins Alone Are Often Incompetent.
Emerging work on organocatalytic enantioselective halocyclizations naturally draws on conditions where both new bonds must be formed under delicate control, the reaction regime where the concerted nature of the AdE3 mechanism is of greatest importance. Without assistance, many simple alkene substrates react slowly or not at all with conventional halenium donors under synthetically relevant reaction conditions. As demonstrated earlier by Shilov, Cambie, Williams, Fahey, and others, alkenes can undergo a concerted AdE3-type reaction via nucleophile participation, which sets the configuration of the newly created stereocenters at both ends in one step. Herein, we explore the modulation of alkene reactivity and halocyclization rates by nucleophile proximity and basicity, through detailed analyses of starting material spectroscopy, addition stereopreferences, isotope effects, and nucleophile-alkene interactions, all obtained in a context directly relevant to synthesis reaction conditions. The findings build on the prior work by highlighting the reactivity spectrum of halocyclizations from stepwise to concerted, and suggest strategies for design of new reactions. Alkene reactivity is seen to span the range from the often overgeneralized "sophomore textbook" image of stepwise electrophilic attack on the alkene and subsequent nucleophilic bond formation, to the nucleophile-assisted alkene activation (NAAA) cases where electron donation from the nucleophilic addition partner activates the alkene for electrophilic attack. By highlighting the factors that control reactivity across this range, this study suggests opportunities to explain and control stereo-, regio-, and organocatalytic chemistry in this important class of alkene additions.


Calbindin-D28K mediates 25(OH)D3/VDR-regulated bone formation through MMP13 and DMP1.
Calcium binding protein calbindin-D28K (CaBP28K) mediates the relationship between vitamin D and calcium, but its mechanism remains unclear during bone formation. The present study reports that maternal CaBP28K levels were positively correlated with paired umbilical cord CaBP28K levels. In addition, CaBP28K levels were positively correlated with the body length, and head and chest circumferences of neonates, but negatively correlated with maternal 25(OH)D3 levels. CaBP28K was also downregulated in MC3T3-E1 osteoblasts when treated with 1,25(OH)2D or VDR overexpression, but was upregulated in the femur of 1α(OH)ase(-/-) mice. Furthermore, it was found CaBP28K may influence cell differentiation and matrix formation through the regulation of DMP1 and the interaction with MMP13 in osteoblasts. This suggests that CaBP28K could be a candidate for the negative role of 1,25(OH)2D/VDR in regulating bone mass.