The Phosphodiesterase Type 5 Inhibitor Sildenafil Improves DNA Stability and Redox Homeostasis in Systemic Sclerosis Fibroblasts Exposed to Reactive Oxygen Species.
Systemic sclerosis (SSc) is a multi-system connective tissue disease characterized by the increased deposition of extracellular matrix proteins such as collagen and fibronectin. Although the pathogenesis is not completely understood, a number of studies suggest that free radicals could be the major contributors to the disease. Indeed, different studies demonstrated how oxidative stress could contribute to the fibrotic process activation at the level of the skin and visceral organs. Emerging evidences highlight the beneficial effects of sildenafil, a phosphodiesterase type 5 inhibitor (PDE5i), which protects different cell lines from the cell damage induced by reactive oxygen species (ROS). These data make sildenafil a good candidate for therapeutic treatment aimed to protect biological macromolecules against oxidative damage, thus preserving cell viability. The purpose of this study was to evaluate the sensitivity of SSc dermal fibroblasts to an oxidative insult and the ability for sildenafil to prevent/reduce the DNA damage due to ROS action. Additionally, we evaluated the capacity for sildenafil to influence redox homeostasis and cytotoxicity, as well as cell proliferation and cell cycle progression. We demonstrated that SSc fibroblasts have an increased sensitivity to a pro-oxidant environment in comparison to healthy controls. The sildenafil treatment reduced ROS-induced DNA damage, counteracted the negative effects of ROS on cell viability and proliferation, and promoted the activity of specific enzymes involved in redox homeostasis maintenance. To our knowledge, in this report, we demonstrate, for the first time, that sildenafil administration prevents ROS-induced instability in human dermal fibroblasts isolated by SSc patients. These results expand the use of PDE5i as therapeutic agents in SSc by indicating a protective role in tissue damage induced by oxidative insult.


Natural course of Rathke's cleft cysts and risk factors for progression.
Rathke's cleft cysts (RCCs) are relatively common and often detected incidentally. They are usually asymptomatic and managed conservatively. However, little is known about their natural history. Thus, the authors aimed to examine the natural course of RCCs and identify the risk factors for their progression. This retrospective study examined 229 patients (median age 43.0 years) diagnosed with RCCs by MRI and followed up without surgery (median period 36.6 months). The median cyst height on the initial MRI was 10 mm. Progression or regression of RCC was defined as cyst height changes of ≥ 1 mm. In total, 23 (10.0%) RCCs progressed, whereas 73 (31.9%) RCCs spontaneously regressed. The remaining 133 were noted to be stable throughout the follow-up period. Patients with progressed RCCs were significantly older than those with stable RCCs. In patients with acute headache as an initial symptom, RCCs were significantly more likely to spontaneously regress. New symptoms occurred in 6 patients, 5 of whom underwent surgery for RCC progression. Of these 6 patients, 1 patient had persistent adrenocorticotropic hormone deficiency and 1 patient developed diabetes insipidus. Kaplan-Meier analysis results showed RCC progression and new symptom development rates to be 12.0% and 4.1% at 5 years and 13.7% and 5.7% at 10 years, respectively. RCCs rarely progress or cause new symptoms in the long term. Patients with asymptomatic RCC should be followed up for at least 5 years to ensure RCC inactivity. RCCs in older adults may require greater surveillance.


Synthesis of a Bent, Twisted, and Chiral Phenanthrene via an Iodine Monochloride-Mediated, Strain-Inducing π-Extension Reaction.
A strategy for the synthesis of substituted and strained p-phenylene units is reported. An oxidative allylic alcohol rearrangement, followed by organometallic addition to the resulting α-ketol and subsequent dehydrative aromatization, affords p-terphenyl-containing macrocycles in which the central p-phenylene has been selectively substituted. Ten 18-membered macrocycles have been synthesized, eight of which contain substituents that could enable π-extension. Only alkynylated derivatives were amenable to π-extension via an ICl-mediated reaction, affording a highly bent, twisted, and chiral phenanthrene.


Imino acids and collagen triple helix stability: characterization of collagen-like polypeptides containing Hyp-Hyp-Gly sequence repeats.
The analysis of factors contributing to the stability of proteins is a subject of intense debate. Particularly challenging is the study of structural proteins, since their function is their structure. Among these is collagen, the key structural component of bones, skin, cartilage, tendons, and other connecting tissues. It is well established that the collagen triple helix is characterized by the presence of hydroxyproline, whose content modulates triple helix thermal stability according to the requirement of the host organism. Because of the complexity and the fibrous nature of collagen, data on the stability and structure of this protein have been mainly obtained by the use of collagen-like polypeptides. On the basis of CD characterization of collagen-like polypeptides we here show that the presence of Hyp at the X position of repeating triplets Hyp-Hyp-Gly stabilizes the triple helix significantly. This extra-stabilization has been ascribed, by using molecular modeling, to the formation of a hydrogen bond between Hyp residues belonging to the X and the Y positions of adjacent chains. This communication also provides a comprehensive interpretation of the ensemble of available data on polypeptides containing proline derivatives.


Ex Vivo Chemosensitivity Profiling of Acute Myeloid Leukemia and Its Correlation With Clinical Response and Outcome to Chemotherapy.
We evaluated the predictive value of the ex-vivo PharmaFlow PM platform in measuring the pharmacological activity of drug combinations consisting of 20 different chemotherapy regimens (20 Tx) administered in 104 acute myeloid leukemia (AML) patients. The predicted sensitivities of alternative treatments for each patient were ranked in five 20% categories, from resistant to sensitive (Groups 1-5). The complete remission (CR) rates of the five groups were 0%, 12.5%, 38.5%, 50.0%, and 81.3%, respectively. The heat map showed a good relationship between drug sensitivity with CR (Group 4 + 5 vs. Group 1 + 2+3: 77.5% vs. 27.3%, p = 0.002) and the European Leukemia Net risk group (22.6% vs. 63.6%, p = 0.015). The predicted coincidence rate was 90.9% in Group 1 + 2 and 81.3% in Group 5. According to the recommendations of the PharmaFlow PM platform, the CR rate would have increased by about 16.3% in one cycle. The overall survival (OS) was shorter in patients predicted to be resistant (Group 1 + 2 vs. Group 3 + 4+5, p = 0.086). In multivariable analysis, CR after one cycle was an independent prognostic factor for OS [p = 0.001; 95% CI 0.202 (0.080-0.511)], and ex-vivo chemosensitivity was a potential predictive factor for OS [p = 0.078; 95% CI 0.696 (0.465-1.041)]. To conclude, the PharmaFlow PM platform is a rapid and valuable tool for predicting clinical response and outcomes in AML patients.


Demonstration of endogenous lectins in synovial tissue.
We have recently shown that synoviocytes and extracellular matrices exhibit distinct patterns of carbohydrate expression. Their biological relevance is however not known. The purpose of the present study was to find out whether human synovial tissue would also show a specific receptor pattern for complex sugar molecules. Endogenous lectins were displayed by means of biotinylated neoglycoproteins and sulfated polysaccharides in paraffin-embedded material or cryosections. In addition to certain carbohydrate components that are known to be constituents of the carbohydrate part of cellular glycoconjugates, our panel included heparin and fucoidan, a sulfated fucose. Binding sites were shown using the avidin-peroxidase technique for light microscopy. The results were compared with immunohistochemical methods and enzyme histochemistry. Our study demonstrates that human synovial tissue contains a complex pattern of endogenous lectins depending on the different types of synovitis. The staining method we used in the investigation allows for precise localization of saccharide binding receptors and is therefore believed to be a reliable technique for further phenotypic characterization of synovial cells.


Case report: passively acquired anti-D in a D+ pregnant patient.
A sample was submitted for serologic evaluation from a pregnant patient with immune thrombocytopenic purpura (ITP) for possible transfusion in the future because of a decreased platelet count. Anti-D and -E were identified in the patient's serum using several antibody identification techniques, and anti-D was recovered in an acid eluate prepared from the patient's red cells. It was discovered that WinRho had been administered to treat the ITP. This product has been licensed for treatment of nonsplenectomized D+ children and adults with ITP to increase the platelet count. Administration of anti-D to D+ individuals for treatment of ITP can cause a red cell anemia.


A Pan-Cancer Analysis of Clinical Prognosis and Immune Infiltration of CKS1B in Human Tumors.
Although more and more evidence supports CDC28 protein kinase subunit 1B (CKS1B) is involved significantly in the development of human cancers, most of the researches have focused on a single disease, and pan-cancer studies conducted from a holistic perspective of different tumor sources have not been reported yet. Here, for the first time, we investigated the potential oncogenic and prognostic role of CKS1B across 33 tumors based on public databases and further verified it in a small scale by RNA sequencing or quantitative real-time PCR. CKS1B was generally highly expressed in a majority of tumors and had a notable correlation with the prognosis of patients, but its prognostic significance in different tumors was not exactly the same. In addition, CKS1B expression was also closely related to the infiltration of cancer-associated fibroblasts in tumors such as breast invasive carcinoma, kidney chromophobe, lung adenocarcinoma, and tumor-infiltrating lymphocytes in tumors such as glioblastoma multiforme, bladder urothelial carcinoma, and brain lower grade glioma. Moreover, reduced CKS1B methylation was observed in certain tumors, for example, adrenocortical carcinoma. Cell cycle and kinase activity regulation and PI3K-Akt signaling pathway were found to be involved in the functional mechanism of CKS1B. In conclusion, our first pan-cancer analysis of CKS1B contributes to a better overall understanding of CKS1B and may provide a new target for future cancer therapy.


Species, biotype and serogroup of Campylobacter spp. isolated from children with diarrhoea over a ten-year period.
From 1981 to 1990, stool samples from 6403 gastroenteritis cases were examined for the presence of campylobacters as well as Salmonella, Shigella, Aeromonas species and Yersinia enterocolitica. The percentages of isolation were the following: campylobacters 10.8 (86.1% of isolates were C. jejuni and 13.9% were C. coli), Salmonella spp. 8.4, Aeromonas spp 1.4, Yersinia enterocolitica 0.3. Shigella spp. were isolated only occasionally. Predominant biotypes of campylobacters were C. jejuni I (69.5%), C. jejuni II (29.5%) and C. coli I (92.7%). The six most common LIO serogroups-36; 4; 1; 28.53; 11; 2-accounted for 50% ca. of typable strains. Campylobacters are the most common etiological agent of bacterial enteritis in children living in this area of Tuscany. The species and serogroup determination can be useful from an epidemiological point of view.


Suppression of antibody responses in vitro by bone marrow cells in rats with hereditary liver injury.
The primary antibody responses of spleen cell cultures were investigated for LEC rats, a mutant strain of LEA rats, which are known to develop spontaneous hepatic injuries. Compared to LEA rats, LEC rats were significantly low in responding not only to a T-dependent antigen, SRBC, but also to a B-cell stimulant, LPS. The antibody responses of LEC rats having fed a diet deprived of Cu metal (LEC/Cu-) showed values intermediate between LEA and LEC rats, thus suggesting a recovery from an immuno-deficient state as well as liver damage in LEC/Cu- rats. The addition of LEA bone marrow cells prior to culture to LEA or LEC spleen cells, strikingly enhanced the antibody response, whereas LEC bone marrow cells were totally ineffective, or sometimes depressed the response. These results have suggested a newer concept for a possible correlation between liver-dysfunction and bone marrow cell-functions in LEC rats, both encoded by autosomal genes.


The phytochrome gene family in tomato and the rapid differential evolution of this family in angiosperms.
A reexamination of the genome of the tomato (renamed Solanum lycopersicum L.) indicates that it contains five, or at most perhaps six, phytochrome genes (PHY), each encoding a different apoprotein (PHY). Five previously identified tomato PHY genes have been designated PHYA, PHYB1, PHYB2, PHYE, and PHYF. A molecular phylogenetic analysis is consistent with the hypothesis that the angiosperm PHY family is composed of four subfamilies (A, B, C/F, and E). Southern analyses indicate that the tomato genome does not contain both a PHYC and a PHYF. Molecular phylogenetic analyses presented here, which utilize for the first time full-length PHY sequences from two completely characterized angiosperm gene families, indicate that tomato PHYF is probably an ortholog of Arabidopsis PHYC. They also confirm that the angiosperm PHY family is undergoing relatively rapid differential evolution. Assuming PHYF is an ortholog of PHYC, PHY genes in eudicots are evolving (Ka/site) at 1.52-2.79 times the rate calculated as average for other plant nuclear genes. Again assuming PHYF is an ortholog of PHYC, the rate of evolution of the C and E subfamilies is at least 1.33 times the rate of the A and B subfamilies. PHYA and PHYB in eudicots are evolving at least 1.45 times as fast as their counterparts in the Poaceae. PHY functional domains also exhibit different evolutionary rates. The C-terminal region of angiosperm PHY (codons 800-1105) is evolving at least 2.11 times as fast as the photosensory domain (codons 200-500). The central region of a domain essential for phytochrome signal transduction (codons 652-712) is also evolving rapidly. Nonsynonymous substitutions occur in this region at 2.03-3.75 times the average rate for plant nuclear genes. It is not known if this rapid evolution results from selective pressure or from the absence of evolutionary constraint.


Determination of ameliorable health impairment influencing health-related quality of life among patients with silicosis in China: a cross-sectional study.
The association between health-related quality of life (HRQOL) and ameliorable health impairment was examined in 208 Chinese in-patients with silicosis, in April 2010. Median St George's Respiratory Questionnaire scores for symptom and activity impairment, social support rating scale and geriatric depression scale scores for patients were 57.24, 59.45, 35 and 5, respectively. Median 36-item short-form health survey physical component (PCS) and mental component (MCS) scores were 47.17 and 51.05, respectively. Lower than median PCS scores (< 47) were significantly associated with high levels of symptom and activity impairment. Lower than median MCS scores (< 51) were significantly associated with high levels of depression and activity impairment. Encouraging patients to participate in daily activities may alleviate activity impairment. Reasonable financial compensation may help to maintain and improve social support levels.


Interdependence of Anion and Cation Exchanges in Calcium Hydroxyapatite: Pb(2+) and Cl(-).
The ion exchange of Pb(2+) and Cl(-) into calcium hydroxyapatite from aqueous solutions is studied at 293 K. Although the exchange of Cl(-) into CaHAp and PbHAp was not observed, in the presence of both Pb(2+) and Cl(-) in aqueous solution the exchange of both Pb(2+) and Cl(-) into CaHAp occurred, and, at least at intermediate concentrations of HCl, a proportionality between the sorbed quantities of these two ions was observed. The dissolution of CaHAp is shown to be dependent upon the pH, while the quantity of Pb(2+) exchanged into the solid is not a function of the proton content; consequently, the exchange is not primarily a dissolution-precipitation process. Copyright 1999 Academic Press.


Odorant receptors of Drosophila are sensitive to the molecular volume of odorants.
Which properties of a molecule define its odor? This is a basic yet unanswered question regarding the olfactory system. The olfactory system of Drosophila has a repertoire of approximately 60 odorant receptors. Molecules bind to odorant receptors with different affinities and activate them with different efficacies, thus providing a combinatorial code that identifies odorants. We hypothesized that the binding affinity of an odorant-receptor pair is affected by their relative sizes. The maximum affinity can be attained when the molecular volume of an odorant matches the volume of the binding pocket. The affinity drops to zero when the sizes are too different, thus obscuring the effects of other molecular properties. We developed a mathematical formulation of this hypothesis and verified it using Drosophila data. We also predicted the volume and structural flexibility of the binding site of each odorant receptor; these features significantly differ between odorant receptors. The differences in the volumes and structural flexibilities of different odorant receptor binding sites may explain the difference in the scents of similar molecules with different sizes.


Early fetal hematopoietic development from in vitro differentiated embryonic stem cells.
In this report we describe the efficient hematopoietic differentiation of embryonic stem (ES) cells in vitro. When cultured in semisolid medium two of five ES cell lines efficiently generated embryoid bodies (EBs) containing blood islands in which hematopoietic cells from all six myeloid lineages could be detected. Among a variety of growth factors tested, only erythropoietin significantly increased blood island formation. We directly demonstrate the presence of hematopoietic progenitors in the EBs by employing an in vitro precursor assay. Colony-forming cells (CFC) of all myeloid lineages as well as bi- and multipotent (CFC-MIX) progenitors were readily identified, and a detailed time-course analysis of their appearance was performed. Despite a high frequency of CFC-MIX in vitro, we did not observe any spleen colony-forming cells (CFU-S) in vivo. We conclude that hematopoietic differentiation of ES cells under these conditions reflects formation of the complete range of blood cells found in the yolk sac of the early fetus. Therefore this system provides a unique model in which to study the earliest events of hematopoietic development in vitro.


Unraveling the mystery of music: music as an evolved group process.
As prominently highlighted by Charles Darwin, music is one of the most mysterious aspects of human nature. Despite its ubiquitous presence across cultures and throughout recorded history, the reason humans respond emotionally to music remains unknown. Although many scientists and philosophers have offered hypotheses, there is little direct empirical evidence for any perspective. Here we address this issue, providing data which support the idea that music evolved in service of group living. Using 7 studies, we demonstrate that people's emotional responses to music are intricately tied to the other core social phenomena that bind us together into groups. In sum, this work establishes human musicality as a special form of social cognition and provides the first direct support for the hypothesis that music evolved as a tool of social living. In addition, the findings provide a reason for the intense psychological pull of music in modern life, suggesting that the pleasure we derive from listening to music results from its innate connection to the basic social drives that create our interconnected world.


Inhibition of Matrix Metalloproteinase-8 Protects Against Sepsis Serum Mediated Leukocyte Adhesion.
Leukocyte adhesion to vascular and matrix Metalloproteinase-8 (MMP8) expression is increased in sepsis and associated with poor prognosis in sepsis patients. This study aimed to investigate the role of MMP8 in sepsis serum mediated leukocyte adhesion. Bioinformatics analysis of GSE64457 and GSE65682 was performed to evaluate the role of MMP8 in the progression of sepsis. Expression of MMP8 in blood samples from patients with sepsis was detected by qRT-PCR and ELISA. Human umbilical vein endothelial cells (HUVECs) were treated with sepsis serum, control serum, and MMP8 inhibitor. Expression of vascular cell adhesion molecule-1 (VCAM-1) and intercellular cell adhesion molecule-1 (ICAM-1) were detected by qRT-PCR and ELISA, respectively. The protein expression of total p38, phosphorylated-p38, ERK1/2, and p-ERK1/2 was detected by Western blotting. Peripheral blood mononuclear cells (PBMCs) and polymorphonuclear neutrophils (PMNs) were incubated with the treated HUVECs to calculate leukocyte adhesion. Four hundred and twenty-nine differentially expressed genes (DEGs) and seven hub genes between sepsis patients and healthy controls were identified. GO function analysis of DEGs and hub genes indicated that the DEGs and hub genes were mainly enriched in neutrophil activation. MMP8 was selected as a key gene with an unfavorable prognosis in sepsis patients. The mRNA and protein expression of MMP8 in blood from sepsis patients were significantly higher than controls. Leukocyte adhesion and mRNA and protein expression of VCAM-1 and ICAM-1 were significantly increased in the sepsis serum group compared to that in the control group, as was the protein expression of p-p38 and p-ERK1/2. However, the MMP8 inhibitor suppressed the leukocyte adhesion promoted by sepsis serum by decreasing the expression of VCAM-1, ICAM-1, p-p38, and p-ERK1/2. Our study indicated that MMP8 acts as a key gene in the development of sepsis, and sepsis serum promotes leukocyte adhesion to HUVECs via MMP8, which suggest that MMP8 might be a potential therapeutic target for sepsis.


Masculinity, femininity, and leadership: Taking a closer look at the alpha female.
An extensive review and textual analysis of the academic and popular literature of the human alpha female was conducted to examine the social construction and expression of the alpha female identity in a small non-random sample of North American women (N = 398). This review revealed 2 predominant alpha female representations in the literature-one more masculine versus one more feminine-and 21 alpha female variables. In this sample of women, the "alpha female" was found to be a recognized socially constructed female identity. Univariate analysis revealed positive and highly significant differences in self-reported mean scores between alpha (N = 94) and non-alpha (N = 304) females for 10 variables including, masculine traits, leadership, strength, low introversion, self-esteem, life satisfaction, sexual experience, initiates sex, enjoys sex and playing a dominant role in sexual encounters, with alpha females scoring higher than non-alphas. The measure of masculine traits was identified as the only predictor of alpha female status as per the multiple regression model. Interestingly, both alpha and non-alpha women scored the same for the measure of feminine traits. Further, both groups scored higher for feminine traits than masculine traits. The results also revealed that neither social dominance nor sexual dominance were predictors of alpha female status which challenge academic and popularized representations of this identity. The results suggest that although the alpha female is often regarded as an exceptional and, at times, an exoticized form of femininity, like other femininities, her identity is marked by contradictions and tensions.


Characterization of Notch receptor expression in the developing mammalian heart and liver.
Alagille syndrome (AGS) is an autosomal dominant disorder characterized by bile duct paucity along with cardiovascular, skeletal, and ophthalmologic defects. The identification of JAG1 as the AGS disease gene revealed the crucial role of the Notch signaling pathway in the development of multiple organ systems in humans. Patients with identical mutations in JAG1 demonstrate extreme clinical variability, suggesting that other factors may influence the severity of the developmental defects in this disorder. We have defined the temporal and spatial expression patterns of the Notch receptor genes in the developing mammalian heart and liver in order to identify potential ligand/receptor interactions during embryogenesis. In the developing heart, both Notch1 and Notch2 are expressed in the outflow tracts and the epicardium, in specific cell populations previously shown to express JAG1. These cells are destined to undergo epithelial-to-mesenchymal transformation. In the newborn mouse liver, Notch2 and Notch3 are expressed in opposing cell populations, suggesting they play different roles in cell fate determination during bile duct development. JAG1 is also expressed in cells adjacent to those expressing Notch2, suggesting a possible ligand receptor relationship. The Notch receptors have distinct roles in cell fate determination in different organ systems.


Case report: tumor-treating fields prolongs IDH-mutant anaplastic astrocytoma progression-free survival and pathological evolution to glioblastoma.
Gliomas are one of the most common tumors of the central nervous system, which cause significant morbidity and mortality. Glioblastoma (GBM) is an aggressive and debilitating disease, which is the most common primary brain tumor in adults. Despite optimal surgical resection followed by radiotherapy and adjuvant chemotherapy, prognosis of patients with GBM remain poor. Tumor-treating fields (TTFields), as a novel treatment approach, is being explored through increased clinical application. Herein, we report the case of a 46-year-old man who was diagnosed as anaplastic astrocytoma [World Health Organization (WHO) III grade], IDH1 R132 mutation, IDH2 R172 mutation, and a methylated MGMT promoter, without 1p36 and 19q13 heterozygosity loss. About 2.5 months after surgery, the patient presented with seizures, aphasia, and memory loss. Following the first-line treatment, TTFields was shown to be a valuable treatment option to control clinical symptom burden. The use of TTFields was shown to prolong progression-free survival and delay the pathological upgradation to glioblastoma. Notably, combined TTFields and chemotherapy might be beneficial in terms of risk reduction for pathological upgradation. To the our current knowledge, this is the first case report that attempts to offer possible strategies based on TTFields toward delaying pathological upgradation from anaplastic astrocytoma (WHO III grade) to glioblastoma.


Role of roots in regulating the growth rate and cytokinin content in leaves.
The role of roots in the enhancement of cytokinin content and leaf growth of Phaseolus vulgaris plants after decapitation and partial defoliation was investigated. Partial excision of the roots of plants which were decapitated above the primary leaf node resulted in a reduction of leaf growth and soluble proteins accumulation in the primary leaves. Roots excision was done at time of decapitation and repeated 8 days later. Endogenous cytokinins, known to be involved in enhancing shoot growth, accumulated in the leaves and stems of decapitated and partially defoliated plants. Lower levels of cytokinins were detected in the leaves of decapitated plants with only a partial root system. The level of cytokinins in the roots of decapitated plants was reduced by partial root excision. The growth and accumulation of cytokinins in leaves were, however, not totally suppressed by removing a large proportion of the roots. At the commencement of the experiment the stem had a higher cytokinin content than both the leaves and roots. This suggests that the stem could be an alternative source of cytokinins to the leaves. The cytokinin complement in the leaves of decapitated plants is not identical to that in the roots. It appears that cytokinins supplied by the roots are metabolized in the leaves, or that alternatively certain cytokinins are synthesized in the leaves themselves.


What are the barriers and enablers that people with a lower limb amputation experience when walking in the community?
Purpose: To explore barriers and enablers to community walking perceived by people with lower limb amputation.Materials and methods: Three focus groups (n = 14 participants), using purposive sampling, were conducted with people with lower limb amputation who used a prosthesis to walk in the community. Three investigators analyzed transcripts using thematic content analysis.Results: Themes aligned with the International Classification of Functioning, Disability and Health domains. Barriers related to body function and structure included prosthetic function, residual limb integrity, phantom limb pain and other medical issues, with optimal prosthetic function and adequate fitness identified as enablers. Personal barriers included challenges adjusting to change, whereas personal enablers included being able to adjust to change, having a positive attitude, goal setting and a purpose for community walking. Environmental barriers included physical (e.g., terrain, crowds, climate) and social (e.g., unwanted attention and finances). Environmental enablers included aids, transport, preparation, social support and finances. Community walking was essential to participation in work, leisure, social activities and family roles.Conclusion: Walking in the community for people with a lower limb amputation presents many barriers, but enablers have also been identified. Targeted assessments and interventions, and appropriate environmental planning may assist in minimizing barriers to community walking.Implications for rehabilitationAlthough rehabilitation for people with lower limb amputation should continue to optimise body function and structure (such as strength, fitness, pain and prosthetic function) to help prepare individuals for walking in the community, personal and environmental factors should also be addressed.Optimising an individual's personal enablers may facilitate community walking; this may include strategies to assist with adjusting to change, setting goals, instilling a positive attitude towards walking, and seeing a purpose for walking in the community.Enhancing environmental (physical and social) enablers and minimising environmental barriers to walking in the community may be possible through environmental planning (e.g. infrastructure such as lifts), social awareness and education.


PRGdb 4.0: an updated database dedicated to genes involved in plant disease resistance process.
The Plant Resistance Genes database (PRGdb; http://prgdb.org/prgdb4/) has been greatly expanded, keeping pace with the increasing amount of available knowledge and data (sequenced proteomes, cloned genes, public analysis data, etc.). The easy-to-use style of the database website has been maintained, while an updated prediction tool, more data and a new section have been added. This new section will contain plant resistance transcriptomic experiments, providing additional easy-to-access experimental information. DRAGO3, the tool for automatic annotation and prediction of plant resistance genes behind PRGdb, has been improved in both accuracy and sensitivity, leading to more reliable predictions. PRGdb offers 199 reference resistance genes and 586.652 putative resistance genes from 182 sequenced proteomes. Compared to the previous release, PRGdb 4.0 has increased the number of reference resistance genes from 153 to 199, the number of putative resistance genes from 177K from 76 proteomes to 586K from 182 sequenced proteomes. A new section has been created that collects plant-pathogen transcriptomic data for five species of agricultural interest. Thereby, with these improvements and data expansions, PRGdb 4.0 aims to serve as a reference to the plant scientific community and breeders worldwide, helping to further study plant resistance mechanisms that contribute to fighting pathogens.


Mutational analysis of the bacteriophage P4 capsid-size-determining gene.
Satellite phage P4 (11,624 bp) depends on the morphopoietic genes (capsid, tail) and lysis genes of its helper phage P2 (33.5 kb) for its lytic development. In the morphopoietic process, P4 redirects the assembly pathway of large, P2 size, capsids (diameter = 60 nm) to yield smaller, P4 size, capsids (diameter = 45 nm), 1/3 in volume of that of its helper. The P4-specified capsid size determination is dependent on the function of the 27-kDa gpSid. To study the capsid size-determining function, we carried out a mutational analysis of the P4 sid gene. Use of a P4-derived genome of 29.1 kb (P461), which can be packaged only into large, P2 size, capsids allowed us to select P4 Sid- mutants. By DNA sequencing we characterized 25 P4 Sid- mutants, of which 10 contain base pair substitutions and 15 contain deletions. Both types of mutations are clustered in separate locations within the sid gene. Our results suggest that the Sid polypeptide contains three distinct functional domains.


Knockdown of long non-coding RNA SNHG8 suppresses the progression of esophageal cancer by regulating miR-1270/BACH1 axis.
The emerging evidence showed that lncRNAs (long non-coding RNAs) could regulate the progression and affect the malignant behaviors of cancers. LncRNA SNHG8 (small nucleolar RNA host gene 8) has been reported to participate in most cancers development. Here in this study, the role of lncRNA SNHG8 in esophageal cancer was uncovered by a series of functional experiments. The expression pattern of SNHG8 in tumor tissues or cells was first detected by qRT-PCR. Using a lentivirus knockdown shRNA is to repress the expression of SNHG8. Subsequently, the in vitro and in vivo experiments were utilized to evaluate whether the malignant behaviors of esophageal cancer were influenced by knockdown SNHG8. The results indicated that lncRNA SNHG8 should be a cancer-promoting factor with a relatively high expression level in esophageal cancer. Moreover, knockdown SNHG8 inhibited the cell viability and induced cell apoptosis in KYSE30 and TE-1 cells. In addition, based on the results of the binding site analysis and the luciferase reporter system, SNHG8 functions by the miR-1270/BACH1 axis. The follow-up experiments verified that lncRNA SNHG8 could down-regulate the expression of miR-1270 to increase the BACH1 expression. Finally, we confirmed that knockdown SNHG8 retarded the progression of esophageal cancer with a xenograft model. To sum up, our findings suggested that lncRNA SNHG8 is a cancer-promoting factor in esophageal cancer. Knockdown SNHG8 could suppress the progression of esophageal cancer, which implies SNHG8 could be used as a therapeutic target in esophageal cancer.


How annual course of photoperiod shapes seasonal behavior of diploid and triploid oysters, Crassostrea gigas.
In this work, we study if ploidy (i.e. number of copies of chromosomes) in the oyster Crassostrea gigas may introduce differences in behavior and in its synchronization by the annual photoperiod. To answer to the question about the effect of the seasonal course of the photoperiod on the behavior of C. gigas according to its ploidy, we quantified valve activity by HFNI valvometry in situ for 1 year in both diploid and triploid oysters. Chronobiological analyses of daily, tidal and lunar rhythms were performed according the annual change of the photoperiod. In parallel, growth and gametogenesis status were measured and spawning events were detected by valvometry. The results showed that triploids had reduced gametogenesis, without spawning events, and approximately three times more growth than diploids. These differences in physiological efforts could explain the result that photoperiod (daylength and/or direction of daylength) differentially drives and modulates seasonal behavior of diploid and triploid oysters. Most differences were observed during long days (spring and summer), where triploids showed longer valve opening duration but lower opening amplitude, stronger daily rhythm and weaker tidal rhythm. During this period, diploids did major gametogenesis and spawning whereas triploids did maximal growth. Differences were also observed in terms of moonlight rhythmicity and neap-spring tidal cycle rhythmicity. We suggest that the seasonal change of photoperiod differentially synchronizes oyster behavior and biological rhythms according to physiological needs based on ploidy.


N-triazinylammonium tetrafluoroborates. A new generation of efficient coupling reagents useful for peptide synthesis.
A new generation of triazine-based coupling reagents (TBCRs), designed according to the concept of "superactive esters", was obtained by treatment of 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium (DMTMM) chloride with lithium or silver tetrafluoroborate. The structure of 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium tetrafluoroborate was confirmed by X-ray diffraction. Activation of carboxylic acids by using this reagent proceeds via triazine "superactive ester". The coupling reagent was successfully used for the synthesis of Z-, Boc-, and Fmoc-protected dipeptides derived from natural and unnatural sterically hindered amino acids and for fragment condensation, in 80-100% yield and with high enantiomeric purity. The manual SPPS of the ACP(65-74) peptide fragment (H-Val-Gln-Ala-Ala-Ile-Asp-Tyr-Ile-Asn-Gly-OH) proceeded significantly faster than with TBTU or HATU, as well as the automated SPPS of the same fragment gave a purer product than by using TBTU or PyBOP. The reagent was also demonstrated to be efficient in on-resin head-to-tail cyclization of constrained cyclopeptides, in SPPS synthesis of Aib peptides, and in the synthesis of esters from appropriate acids, alcohols, and phenols. The high efficiency and versatility of this new generation of TBCRs confirm, for the first time, the usefulness of the concept of "superactive esters" in rational design of the structure of coupling reagents.


[Posterior uveitis in a 47-year old patient infected with HIV--case report].
To report the case of posterior uveitis associated with HIV infection and to discuss contemporary references on the topic. A 47-years old woman with asymptomatic HIV infection developed acute posterior uveitis. The examinations for infections with CMV, Toxoplasma gondii, Treponema pallidum and Mycobacteria were negative. After failure of empirical treatment with antibiotics, ganciclovir and steroids antiretroviral treatment with three drugs was initiated on the patient. The ophthalmologic examination disclosed significant improvement after two weeks of the treatment. There have been no evidences of inflammatory disease in the eyes for 12 weeks from the starting of antiretroviral therapy. The immunologic and virologic parameters showed good control of HIV infection. Infection with HIV appears to be a cause of uveitis.


Macrophages of the subcutaneous and omental fatty tissue in obese patients: Immunohistochemical phenotyping of M2 subtypes in relation to type 2 diabetes.
Macrophages are linked to the initiation of the chronic inflammation believed to underlie the changes taking place in the white fatty tissue of obese people. Both the number of macrophages, but their functional status, play an important role in the development of inflammation. Classically, macrophages are divided into two types: pro-inflammatory (M1) and anti-inflammatory (M2) types, and based on current immunological studies, further views on the functional distribution of macrophages are suggested. In this study, we evaluated the M1 and M2 macrophages ratio in obese subjects with, or without diabetes. To identify all macrophages, we used CD68 expression, while CD204 expression is typically used for the M2 macrophage. During bariatric surgery, carried out in obese people with and without type 2 diabetes (T2D), we obtained subcutaneous adipose tissue from the navel and omental adipose tissue. We also obtained the same tissue from people with a physiological range of BMI from a judicial autopsy. Applying immunohistochemical staining anti-CD68 and anti-CD204, we carried out a quantitative evaluation of the number of macrophages. We found CD68+ and CD204+ positive macrophages in perivascular spaces and between fat cells, both isolated and in larger infiltrates. They were also present in so-called "crown-like structures" (CLS) around dying adipocytes. Quantitative analysis showed an increased number of macrophages in all obese patients compared to the control group of non-obese, individuals without T2D. The most striking observation was the macrophage increase in the visceral fatty tissue of diabetics. The number of CD68 and CD204 positive macrophages was statistically significantly smaller in patients without T2D. We demonstrated a significantly greater number of macrophages in visceral adipose tissue, especially in patients with T2D. Our results also show a positive correlation between the presence of T2D and the total number of macrophages; a significantly greater number of macrophages were found in visceral adipose tissue, especially in patients with T2D.


Accuracy for diagnosis of periapical cystic lesions.
Clinical differentiation between cystic lesions of endodontic and non-endodontic origin is of importance because correct diagnosis may affect treatment decision making. Most radicular cysts are treated with conservative approaches and, therefore, are not surgically removed. The objective of this study was to determine the accuracy of clinical diagnosis of periapical lesions as compared to the histological findings, and to evaluate various associated factors. All biopsy specimens submitted for histological evaluation from 2002 to 2009 were assessed. Only cases of periapical lesions with complete patient data and clinical diagnosis were included. Sensitivity, specificity and accuracy of the clinical diagnosis were calculated and various patient-related factors were evaluated. Of the 4,908 cases, 183 met inclusion criteria. Histologically, there were 171 lesions of radicular cysts and 12 cases of non-endodontic cysts, including OKC and Incisive Canal Cyst. The diagnostic accuracy for clinical diagnosis for radicular cysts was 91.84% and 91.84% for non-endodontic cysts. There was a high accuracy of clinical differentiation between cystic lesions of endodontic and non-endodontic origin. However, some non-endodontic lesions may be incorrectly diagnosed clinically as lesions of endodontic origin. Histological evaluation may be necessary for the correct diagnosis. Further clinical studies are needed to evaluate clinical examination and histological diagnosis of periapical lesions.


[The presence of large acidic vesicles is correlated with in vitro invasive potential of breast cancer cells].
Human breast cancer cells include large acidic vesicles (LAVs) containing endocytosed extracellular matrix (P. Montcourrier et al., Cancer Res., 50, 1990, p. 6045-6054) and mature cathepsin D, a lysosomal protease associated with metastatic risk. We show that metastatic breast cancer MDA-MB231 cells migrating in vitro through a Matrigel gel in a modified Boyden chamber were enriched in LAVs (28% compared to 6% before plating) as visualized by acridine orange fluorescence. This is the first evidence that LAVs are associated with invasiveness of cancer rather than with an apoptotic process.


Oral frostbite injury from intentional abuse of a fluorinated hydrocarbon.
A serious but rarely reported complication of halogenated hydrocarbon inhalation abuse is severe mucosal frostbite. A 16-year-old male attempted to "get high" by inhaling airbrush propellant which contained 1,1-difluoroethane (CAS #75-376). The patient lost consciousness and upon awakening his lips and tongue were frozen. He suffered first- and second-degree burns of the larynx with vocal cord involvement and first-degree burns of the trachea, main stem bronchi, and esophagus. The oral cavity had second- and third-degree burns which required debridement. This case demonstrates the unusual but severe damage that can occur with the abuse of fluorinated hydrocarbons.


[cDNA library construction from panicle meristem of finger millet].
The protocol for production of full-size cDNA using SuperScript Full-Length cDNA Library Construction Kit II (Invitrogen) was tested and high quality cDNA library from meristematic tissue of finger millet panicle (Eleusine coracana (L.) Gaertn) was created. The titer of obtained cDNA library comprised 3.01 x 10(5) CFU/ml in avarage. In average the length of cDNA insertion consisted about 1070 base pairs, the effectivity of cDNA fragment insertions--99.5%. The selective sequencing of cDNA clones from created library was performed. The sequences of cDNA clones were identified with usage of BLAST-search. The results of cDNA library analysis and selective sequencing represents prove good functionality and full length character of inserted cDNA clones. Obtained cDNA library from meristematic tissue of finger millet panicle represents good and valuable source for isolation and identification of key genes regulating metabolism and meristematic development and for mining of new molecular markers to conduct out high quality genetic investigations and molecular breeding as well.


Foodborne proportion of gastrointestinal illness: estimates from a Canadian expert elicitation survey.
The study used a structured expert elicitation survey to derive estimates of the foodborne attributable proportion for nine illnesses caused by enteric pathogens in Canada. It was based on a similar study conducted in the United States and focused on Campylobacter, Escherichia coli O157:H7, Listeria monocytogenes, nontyphoidal Salmonella enterica, Shigella spp., Vibrio spp., Yersinia enterocolitica, Cryptosporidium parvum, and Norwalk-like virus. For each pathogen, experts were asked to provide their best estimate and low and high limits for the proportion of foodborne illness relative to total cases. In addition, they provided background information with regard to food safety experience, including self-evaluated expertise for each pathogen on a 5-point scale. A snowball approach was used to identify 152 experts within Canada. The experts' background details were summarized using descriptive statistics. Factor analysis was used to determine whether the variability in best estimates was related to self-assessed level of expertise or other background information. Cluster analysis followed by beta function fitting was undertaken on best estimates from experts who self-evaluated their expertise 3 or higher. In parallel, Monte Carlo resampling was run using triangular distributions based on each expert's best estimate and its limits. Sixty-six experts encompassing various academic backgrounds, fields of expertise, and experiences relevant to food safety provided usable data. Considerable variation between experts in their estimated foodborne attributable proportions was observed over all diseases, without any relationship to the expert's background. Uncertainty about their estimate (measured by the low and high limits) varied between experts and between pathogens as well. Both cluster analysis and Monte Carlo resampling clearly indicated disagreement between experts for Campylobacter, E. coli O157, L. monocytogenes, Salmonella, Vibrio, and Y. enterocolitica. In the absence of more reliable estimates, the observed discrepancy between experts must be explored and understood before one can judge which opinion is the best.


Macrophage polarization and allergic asthma.
Allergic asthma is associated with airway inflammation and airway hyperresponsiveness. Macrophage polarization has been shown to have a profound impact on asthma pathogenesis. On exposure to local microenvironments, recruited macrophages can be polarized into either classically activated (or M1) or alternatively activated (or M2) phenotypes. Macrophage polarization has been heavily associated with development of asthma. The process of regulation of macrophage polarization involves an intricate interplay between various cytokines, chemokines, transcriptional factors, and immune-regulatory cells. Different signals from the microenvironment are controlled by different receptors on the macrophages to initiate various macrophage polarization pathways. Most importantly, there is an increased attention on the epigenetic changes (eg, microRNAs, DNA methylation, and histone modification) that impact macrophage functional responses and M1/M2 polarization through modulating cellular signaling and signature gene expression. Thus, modulation of macrophage phenotypes through molecular intervention by targeting some of those potential macrophage regulators may have therapeutic potential in the treatment of allergic asthma and other allergic diseases. In this review, we will discuss the origin of macrophages, characterization of macrophages, macrophage polarization in asthma, and the underlying mechanisms regarding allergen-induced macrophage polarization with emphasis on the regulation of epigenetics, which will provide new insights into the therapeutic strategy for asthma.


Cost-effectiveness of high-dose intravenous esomeprazole in patients with peptic ulcer bleeding in the USA and Europe.
Peptic ulcer bleeding (PUB) is life-threatening and associated with high healthcare costs. Clinical outcomes in PUB depend largely on the risk of rebleeding. Recent data indicate that intravenous proton pump inhibitors (PPIs) reduce rebleeds, the need for surgery and repeat endoscopic treatment. From a policy perspective, it is important to assess the cost-effectiveness of this treatment. Accordingly, a decision-tree model published by Barkun et al. evaluated the costs and benefits of high-dose intravenous esomeprazole in preventing rebleeds in patients with PUB based on data from a multinational, randomized clinical trial comparing this therapeutic approach to intravenous placebo. The results indicate that esomeprazole is cost effective in the USA and Sweden, and cost saving in Spain. These findings agree with most other analyses of intravenous PPIs used in PUB patients at high risk for bleeds. The therapeutic approach provides increased benefits to patients at a relatively small additional cost.


Own-Group Face Recognition Bias: The Effects of Location and Reputation.
In the present study, we examined whether social categorization based on university affiliation can induce an advantage in recognizing faces. Moreover, we investigated how the reputation or location of the university affected face recognition performance using an old/new paradigm. We assigned five different university labels to the faces: participants' own university and four other universities. Among the four other university labels, we manipulated the academic reputation and geographical location of these universities relative to the participants' own university. The results showed that an own-group face recognition bias emerged for faces with own-university labels comparing to those with other-university labels. Furthermore, we found a robust own-group face recognition bias only when the other university was located in a different city far away from participants' own university. Interestingly, we failed to find the influence of university reputation on own-group face recognition bias. These results suggest that categorizing a face as a member of one's own university is sufficient to enhance recognition accuracy and the location will play a more important role in the effect of social categorization on face recognition than reputation. The results provide insight into the role of motivational factors underlying the university membership in face perception.


Analgesic effects induced by TENS and electroacupuncture with different types of stimulating electrodes on deep tissues in human subjects.
Effects of conditioning peripheral nerve stimulation with different types of stimulating electrodes on pain thresholds in various deep tissues were measured in human subjects. Cone-shaped metal (phi 13 mm), rubber (phi 13 mm), and large soft surface electrodes (50 x 150 mm) were used for transcutaneous electrical nerve stimulation (TENS), and insulated and non-insulated acupuncture needles (diameter: 240 microns) were used for electroacupuncture (EA). Two pairs of electrodes were placed around the point of deep pain measurement. Symmetrical positive and negative square pulses (0.1 msec at 100 Hz) of just below the pain tolerance intensity were used for both TENS and EA. Deep pain thresholds were measured at the center of the thigh with a pulse algometer and insulated needle electrodes. Pain thresholds of deep tissues were in the order periosteum < fascia < skin (including subcutaneous tissues) < muscle. TENS with surface electrodes significantly increased pain thresholds of skin and fascia but not those of muscle or periosteum. The shape, material and size of the surface electrodes hardly affected the degree of analgesic effect, except in the fascia by large soft electrodes. In contrast, EA with non-insulated needles induced a greater increase in pain threshold in skin, fascia and muscle, although statistically significant results were obtained in only the first two tissues. EA with insulated needle electrodes was the only technique with which we obtained a significant increase in pain threshold in muscle and periosteum. These results suggest that the choice of electrode and stimulus parameters is important for the production of sufficient analgesic effects in different somatic tissues and that insulated needle electrodes are useful for pain relief in deeper tissues such as muscle and periosteum.


Enhancing collaborative filtering by user interest expansion via personalized ranking.
Recommender systems suggest a few items from many possible choices to the users by understanding their past behaviors. In these systems, the user behaviors are influenced by the hidden interests of the users. Learning to leverage the information about user interests is often critical for making better recommendations. However, existing collaborative-filtering-based recommender systems are usually focused on exploiting the information about the user's interaction with the systems; the information about latent user interests is largely underexplored. To that end, inspired by the topic models, in this paper, we propose a novel collaborative-filtering-based recommender system by user interest expansion via personalized ranking, named iExpand. The goal is to build an item-oriented model-based collaborative-filtering framework. The iExpand method introduces a three-layer, user-interests-item, representation scheme, which leads to more accurate ranking recommendation results with less computation cost and helps the understanding of the interactions among users, items, and user interests. Moreover, iExpand strategically deals with many issues that exist in traditional collaborative-filtering approaches, such as the overspecialization problem and the cold-start problem. Finally, we evaluate iExpand on three benchmark data sets, and experimental results show that iExpand can lead to better ranking performance than state-of-the-art methods with a significant margin.


Fiber Bragg gratings inscribed using 800nm femtosecond laser and a phase mask in single- and multi-core mid-IR glass fibers.
For the first time, Fiber Bragg grating (FBG) structures have been inscribed in single-core passive germanate and three-core passive and active tellurite glass fibers using 800 nm femtosecond (fs) laser and phase mask technique. With fs peak power intensity in the order of 10(11)W/cm(2), the FBG spectra with 2nd and 3rd order resonances at 1540 and 1033 nm in the germanate glass fiber and 2nd order resonances at approximately 1694 and approximately 1677 nm with strengths up to 14 dB in all three cores in the tellurite fiber were observed. Thermal responsivities of the FBGs made in these mid-IR glass fibers were characterized, showing average temperature responsivity approximately 20 pm/ degrees C. Strain responsivities of the FBGs in germanate glass fiber were measured to be 1.219 pm/microepsilon.


Concentration of Indole-3-acetic Acid and Its Derivatives in Plants.
Seeds of oat, coconut, soybean, sunflower, rice, millet, kidney bean, buckwheat, wheat, and corn and vegetative tissue of oat, pea, and corn were assayed for free indole-3-acetic acid (IAA), esterified IAA, and peptidyl IAA. Three conclusions were drawn: (a) all plant tissues examined contained most of their IAA as derivatives, either esterified or as a peptide; (b) the cereal grains examined contained mainly ester IAA; (c) the legume seeds examined contained mainly peptidyl IAA. Errors in analysis of free and bound IAA are discussed.


ETV6-PDGFRB and FIP1L1-PDGFRA stimulate human hematopoietic progenitor cell proliferation and differentiation into eosinophils: the role of nuclear factor-κB.
ETV6-PDGFRB (also called TEL-PDGFRB) and FIP1L1-PDGFRA are receptor-tyrosine kinase fusion genes that cause chronic myeloid malignancies associated with hypereosinophilia. The aim of this work was to gain insight into the mechanisms whereby fusion genes affect human hematopoietic cells and in particular the eosinophil lineage. We introduced ETV6-PDGFRB and FIP1L1-PDGFRA into human CD34(+) hematopoietic progenitor and stem cells isolated from umbilical cord blood. Cells transduced with these oncogenes formed hematopoietic colonies even in the absence of cytokines. Both oncogenes also stimulated the proliferation of cells in liquid culture and their differentiation into eosinophils. This model thus recapitulated key features of the myeloid neoplasms induced by ETV6-PDGFRB and FIP1L1-PDGFRA. We next showed that both fusion genes activated the transcription factors STAT1, STAT3, STAT5 and nuclear factor-κB. Phosphatidylinositol-3 kinase inhibition blocked nuclear factor-κB activation in transduced progenitor cells and patients' cells. Nuclear factor-κB was also activated in the human FIP1L1-PDGFRA-positive leukemia cell line EOL1, the proliferation of which was blocked by bortezomib and the IκB kinase inhibitor BMS-345541. A mutant IκB that prevents nuclear translocation of nuclear factor-κB inhibited cell growth and the expression of eosinophil markers, such as the interleukin-5 receptor and eosinophil peroxidase, in progenitors transduced with ETV6-PDGFRB. In addition, several potential regulators of this process, including HES6, MYC and FOXO3 were identified using expression microarrays. We show that human CD34(+) cells expressing PDGFR fusion oncogenes proliferate autonomously and differentiate towards the eosinophil lineage in a process that requires nuclear factor-κB. These results suggest new treatment possibilities for imatinib-resistant myeloid neoplasms associated with PDGFR mutations.


The mundane realities of the everyday lay use of the internet for health, and their consequences for media convergence.
The internet is now a major source of health information for lay people. Within the medical, sociological and popular literatures there have been three main responses to this development. We classify these as "celebratory", "concerned" and "contingent". This paper falls into the third category and, drawing on techniques of discourse analysis, examines people's accounts of their use of online health resources. It identifies six implicit rules--which we call "rhetorics of reliability"--that people readily draw upon when articulating why they trust some online sources and not others. In addition participants locate their accounts within broader discursive frameworks in order to present themselves as "sensible" users. The article concludes by suggesting that there is an emerging concordance between the lay use of the internet for health and illness and dominant (generally) biomedical conceptions of what constitutes "good quality" health information.


Imaging the spine.
This article on neuroradiology concerns the imaging of the spine. The imaging modalities are discussed, and some basic imaging strategies are outlined.


Conservation of extrusion as an exit mechanism for Chlamydia.
Chlamydiae exit via membrane-encased extrusion or through lysis of the host cell. Extrusions are novel, pathogen-containing structures that confer infectious advantages to Chlamydia, and are hypothesized to promote cell-to-cell spread, dissemination to distant tissues and facilitate immune evasion. The extrusion phenomenon has been characterized for several Chlamydia trachomatis serovars, but a thorough investigation of extrusion for additional clinically relevant C. trachomatis strains and Chlamydia species has yet to be performed. The key parameters investigated in this study were: (i) the conservation of extrusion across the Chlamydia genus, (ii) the functional requirement for candidate Chlamydia genes in extrusion formation i.e. IncA and CT228 and (iii) extrusion-mediated uptake, and consequent survival of Chlamydia inside macrophages. Inclusion morphology was characterized by live fluorescence microscopy, using an inverted GFP strategy, at early and mid-stages of infection. Enriched extrusions were used to infect bone marrow-derived macrophages, and bacterial viability was measured following macrophage engulfment. Our results demonstrate that extrusion is highly conserved across chlamydiae, including ocular, STD and LGV biovars and divergent Chlamydia species. Consequently, this exit mechanism for Chlamydia may fulfill common advantages important for pathogenesis.


Myocarditis masquerading as ischemic heart disease: the diagnostic utility of antimyosin imaging.
The diagnosis of myocarditis presents a diagnostic challenge due to its varied clinical presentation. In addition, criteria for myocarditis are varied. At present, the confirmation of myocarditis depends on an endomyocardial biopsy demonstrating myocardial inflammation and necrosis. Unfortunately, this invasive procedure is associated with some degree of risk and has significant limitations. This report discusses the case presentations of two patients with chest pain, electrocardiographic changes and elevated creatine kinase levels suggestive of myocardial infarction, who were subsequently found to have findings compatible with myocarditis based on indium-111 antimyosin antibody scanning. This noninvasive test therefore appears to have value in the differentiation of myocardial ischemia from myocarditis.


Ocular surface microbiota: Ophthalmic infectious disease and probiotics.
Recently, increasing studies have emphasized the importance of commensal bacteria in humans, including microbiota in the oral cavity, gut, vagina, or skin. Ocular surface microbiota (OSM) is gaining great importance as new methodologies for bacteria DNA sequencing have been published. This review outlines the current understanding and investigation of OSM and introduces the new concept of the gut-eye axis. Moreover, we have collected current studies that focus on the relationship between ophthalmic infectious disease and alterations in the OSM or human gut microbiota. Finally, we discuss the current application of probiotics in ophthalmic infectious disease, its limitations to date, and futural directions.


Upper eyelid extension of a filtering bleb following glaucoma shunt surgery.
The authors describe a case of a filtering bleb that extended into the eyelid of a 71-year-old man following glaucoma surgery with a shunt implant. The patient presented with a left upper eyelid mass and had ocular surface and mechanical sequelae. Computed tomography scan demonstrated a fistula between the original filtering tract and the eyelid, creating an inadvertent filtering bleb. Cytology revealed fluid consistent with aqueous humor. Intraocular pressure remained normal and symptoms improved with conservative management. The patient deferred any surgical revision. Glaucoma tube shunts may lead to ocular and rare orbital complications from inadvertent bleb extension, as seen in this case.


The rich-get-richer effect: Prior knowledge predicts new learning of domain-relevant information.
When learning new information, students' prior knowledge related to that information will often vary. Prior research has not systematically explored how prior knowledge relates to learning of new, previously unknown information. Accordingly, the goal of the present research was to explore this relationship. In three experiments, students first completed a prior knowledge test over two domains (football and cooking) and then learned new information from these domains by answering questions and receiving feedback. Students also made a judgment of learning for each. To ensure that the learning was new (i.e., previously unknown) for all students, the to-be-learned information was false. Last, students completed a final test over the same questions from the learning phase. Prior knowledge in each domain was positively related to new learning for items from that domain but not from the other domain. Thus, the relationship between prior knowledge and new learning was domain specific, which we refer to as the rich-get-richer effect. Prior knowledge was also positively related to the magnitude of judgments of learning. In Experiment 3, to explore a potential reason why prior knowledge is related to new learning, students rated their curiosity in learning each item prior to receiving feedback. Critically, students' curiosity judgments mediated the relationship between prior knowledge and new learning. These outcomes suggest that for high-knowledge learners, curiosity may be related to attention-based mechanisms that increase the effectiveness of encoding during feedback. (PsycInfo Database Record (c) 2022 APA, all rights reserved).


A qualitative study of perceived needs and factors associated with the quality of care for common mental disorders in patients with chronic diseases: the perspective of primary care clinicians and patients.
The prevalence of comorbid anxiety and depressive disorders is high among patients with chronic diseases in primary care, and is associated with increased morbidity and mortality rates. The detection and treatment of common mental disorders in patients with chronic diseases can be challenging in the primary care setting. This study aims to explore the perceived needs, barriers and facilitators for the delivery of mental health care for patients with coexisting common mental disorders and chronic diseases in primary care from the clinician and patient perspectives. In this qualitative descriptive study, we conducted semi-structured interviews with clinicians (family physician, nurse, psychologist, social worker; n = 18) and patients (n = 10) from three primary care clinics in Quebec, Canada. The themes explored included clinician factors (e.g., attitudes, perception of roles, collaboration, management of clinical priorities) and patient factors (e.g., needs, preferences, access to care, communication with health professionals) associated with the delivery of care. Qualitative data analysis was conducted based on an interactive cyclical process of data reduction, data display and conclusion drawing and verification. Clinician interviews highlighted a number of needs, barriers and enablers in the provision of patient services, which related to inter-professional collaboration, access to psychotherapy, polypharmacy as well as communication and coordination of services within the primary care clinic and the local network. Two specific facilitators associated with optimal mental health care were the broadening of nurses' functions in mental health care and the active integration of consulting psychiatrists. Patients corroborated the issues raised by the clinicians, particularly in the domains of whole-person care, service accessibility and care management. The results of this project will contribute to the development of quality improvement interventions to increase the uptake of organizational and clinical evidence-based practices for patients with chronic diseases and concurrent common mental disorders, in priority areas including collaborative care, access to psychotherapy and linkages with specialized mental health care.


Role of SraP in adherence of Staphylococcus aureus to the bovine mammary epithelia.
SraP, a platelet-binding surface protein of Staphylococcus aureus, is involved in the pathogenesis of infective endocarditis. In this study, we investigated the importance of SraP in the pathogenesis of bovine mastitis. By means of PCR, sraP was detected in all the isolates tested from bovine bulk milk and humans. However, SraP was not expressed on the cell surface in half of the bovine isolates. Moreover, disruption of sraP did not affect the ability of S. aureus to adhere to cultured bovine mammary epithelial cells. These results suggest that SraP does not seem to be an important factor for S. aureus to adhere to the bovine mammary epithelia.


Diffuse idiopathic skeletal hyperostosis in a young woman treated with isotretinoin.
We present the case of a 36-year-old woman with diffuse idiopathic skeletal hyperostosis especially at cervical spine since she was 29 years old. The only relevant feature was the use of isotretinoin at regular doses in the past for severe acne.


Pathologic prognostic factors in the recurrence of rectal cancer.
For rectal cancer, local recurrence following surgical treatment is a grave complication that occurs in as many as 25% of cases. Pathological examination of the surgical resection specimen plays a primary role in assessing both the surgery- and tumor-related factors that contribute to the risk of recurrence. Among the tumor-related factors, stage has long been considered the single most accurate indicator of survival. However, recent evidence strongly suggests that the most powerful predictor of both local recurrence and overall outcome in the absence of distant metastatic disease is the macroscopic quality of the mesorectum in the resection specimen and the proximity of the tumor to the circumferential (radial) resection margin. Additional pathologic features have been shown to have stage-independent prognostic significance in colorectal cancer and may help to further define risk of adverse outcome. Such features include: tumor grade; histologic type; extent of extramural penetration by tumor; neural, venous, and/or lymphatic invasion; tumor border configuration; tumor budding; and host lymphoid response. The predictive value of tumor-specific molecular features is currently under investigation and may help to further improve prognostication and refine individual patient management in rectal cancer.


Distribution and retention of cadmium in metallothionein I and II null mice.
Cadmium (Cd) is known to have a long biological half-life in the body, possibly due to its binding to metallothionein (MT). This study was designed to determine the role of MT in the tissue distribution and retention of Cd using MT-I and -II null (MT-null) mice. Mice were given 109CdCl(2) (15 mumol/kg, 25 microCi/kg, i.p.), and radioactivity was quantified in 14 major organs at 2 hr, 1, 2, 3, 7 and 15 days thereafter. The lack of MT in MT-null mice 2 hr after Cd administration (74% vs 72% of the dose, respectively). However, the elimination of Cd was much faster in MT-null mice than in control mice. In control mice, approximately 40% of Cd administered was found in liver 24 hr after administration, and the majority was bound to MT. In contrast, only 20% of Cd was found in liver of MT-null mice, which was not bound to MT. Cd concentrations in kidney, pancreas, and spleen were also lower in MT-null than in control mice 1 week after administration. No apparent difference in Cd retention in other organs was noted between control and MT-null mice over the 15-day period. Cd concentration in kidney continued to increase with time in control but not in MT-null mice, indicating that an important source of Cd in the kidney is the uptake of CdMT. In conclusion, the present data indicate that MT does not play a role in the initial distribution of Cd to tissues, but does play a major role in the elimination of Cd, especially from liver, kidney, and pancreas. These data support the conclusion that the persistence of Cd in the body is at least partially due to Cd binding to MT in tissues.


Evaluation of hybridization characteristics of a cloned pRF106 probe for Listeria monocytogenes detection and development of a nonisotopic colony hybridization assay.
An internal fragment (pRF106 fragment, ca. 500 bp) of a gene (msp) coding for a 60-kDa protein of Listeria monocytogenes serotype 1/2a was used to develop a screening method to discriminate between L. monocytogenes and avirulent Listeria spp. on primary isolation plates. The L. monocytogenes-derived probe fragment of pRF106 hybridized to a 13-kb fragment of L. monocytogenes and a 3-kb fragment of one cheese isolate strain of Listeria seeligeri under stringent hybridization conditions (mean thermal denaturation temperature [Tm]-5 degrees C). The probe also hybridized to a 6-kb fragment of Listeria innocua, Listeria ivanovii, and L. seeligeri under less stringent hybridization conditions (Tm-17 degrees C). The pRF106 fragment was labeled with digoxigenin-11-dUTP and used to develop a colony hybridization assay. Colonies from lithium chloride-phenylethanol-moxalactam agar were blotted onto nylon membranes. The cells were pretreated with microwaves before lysis with sodium hydroxide. DNA-DNA hybridization and posthybridization washing were done at high stringency (Tm-7 degrees C). The nonisotopic colony hybridization procedure was specific for L. monocytogenes when evaluated against pure cultures of L. monocytogenes and other Listeria species, excluding the cheese isolate of L. seeligeri. Also, it was specific for L. monocytogenes when evaluated with Listeria-negative food enrichment cultures that were inoculated in the laboratory with Listeria species.


Effect of weather on Ips typographus (Coleoptera Curculionidae) phenology, voltinism, and associated spruce mortality in the southeastern Alps.
Summer drought associated with high temperatures recorded in the last few years has given rise to outbreaks of bark beetles developing in weakened host trees. The aim of this study was to investigate the possible weather effect on the biology of and damage caused by Ips typographus L. in the southeastern Alps. The study was carried out recording temperature (1962-2007), precipitation (1922-2007), and the damage caused by I. typographus (1993-2007). In addition, data from pheromone-baited traps (1996-2005) provided information on the main periods of flight activity of I. typographus. From 1922 to 2007, precipitation during March-July has decreased approximately 200 mm (-22%), whereas since 1962-2007, mean temperatures during March-July increased approximately 2 degrees C (+13%). Damage caused by I. typographus was inversely correlated with March-July precipitation from the previous year but not correlated with temperature. Increases in spring temperature did not affect the development timing of the first generation, but only changed its onset. Earlier swarming of both overwintering beetles and first-generation offspring ( approximately 20 d sooner over 10 yr), and the early start of the second generation permitted more complete development of the second brood. Voltinism in this species is discussed in relation to thermal and photoperiodic thresholds, indicating that the occurrence of a third generation is limited by the summer photoperiod rather than by temperature. In conclusion, results suggest that spring drought increases damage caused by I. typographus in the following year, whereas warmer spring affects insect phenology.


Use of structure-based drug design approaches to obtain novel anthranilic acid acyl carrier protein synthase inhibitors.
Acyl carrier protein synthase (AcpS) catalyzes the transfer of the 4'-phosphopantetheinyl group from the coenzyme A to a serine residue in acyl carrier protein (ACP), thereby activating ACP, an important step in cell wall biosynthesis. The structure-based design of novel anthranilic acid inhibitors of AcpS, a potential antibacterial target, is presented. An initial high-throughput screening lead and numerous analogues were modeled into the available AcpS X-ray structure, opportunities for synthetic modification were identified, and an iterative process of synthetic modification, X-ray complex structure determination with AcpS, biological testing, and further modeling ultimately led to potent inhibitors of the enzyme. Four X-ray complex structures of representative anthranilic acid ligands bound to AcpS are described in detail.


Expression and characterization of recombinant alpha-galactosidase in baculovirus-infected insect cells.
A cDNA encoding coffee bean alpha-galactosidase was subcloned into baculovirus expression vectors, pVL-1393 and pAc-GP67B, for intracellular and extracellular expression in Spodoptera frugiperda (Sf9) insect cells, respectively. The expressed protein (recombinant alpha-galactosidase) was immunologically reactive with antisera raised against its native counterpart isolated from coffee beans and was biologically active towards the substrate p-nitrophenyl alpha-galactopyranoside. The subcellular distribution of recombinant alpha-galactosidase expressed from different vectors was analyzed by Western blotting, immunofluorescent labeling, and electron microscopy. In addition, recombinant alpha-galactosidase was compared to the native enzyme with respect to glycosylation, thermostability, and pH profile. Furthermore, a recombinant alpha-galactosidase molecule with a His6 tag at its C-terminus was constructed by an overlap PCR method so that the enzyme expressed in Sf9 cells can be purified by a simple affinity chromatography procedure.


[Localization of glutathione-S-transferase in transparent and cataractous human lenses].
The localization of glutathione-S-transferase (GST) activity was investigated in 15 cataractous lenses obtained by intracapsular extraction of senile cataract. Additionally, 8 clear lenses, obtained from donor eyes in cases of corneal transplantation or of traumatic lens luxation, were used as controls. The lenses were divided into the central and peripheral portions with a trephine. The activity of GST was quantitated in each portion according to the method described by Habig. GST activity in clear lenses was significantly higher in the peripheral portion than in the central portion and in the former it was significantly higher in clear lenses than in cataractous ones. The ratio of GST activity between central and peripheral portions (C/P) was significantly higher in cataractous lenses than in clear ones. These findings suggested that the scavenging system of the human clear lens is more active in the equatorial portion than in the central portion. Nevertheless the activity of GST in human cataractous lenses decreased more significantly in the peripheral portion than in the central portion. These findings indicated that the human cataract usually develop from equatorial portion of lens.


Retinopathy in a Full-Term Infant with Cri-du-Chat Syndrome.
Cri-du-chat (CdC) is a 5p chromosomal deletion syndrome. CdC has numerous systemic associations but only a few ocular manifestations have been documented. In this report we present novel ocular findings of peripheral avascular retina and retinal hemorrhages in a full-term female infant, born to non-consanguineous parents, who had clinical features of cri-du-chat syndrome and genetic confirmation. The retinal hemorrhages resolved. However, the temporal avascular retina in our full-term patient remained. Further analysis of the 5p locus showed 3 genes: CTNND2, SEMA5A and SLC6A18 that not only fit our patient's external phenotype and ophthalmoscopic findings of retinal hemorrhages, but were also key in proper ocular development and neurogenesis, suggesting a genetic contribution by the short-arm of chromosome 5 to proper retinal maturation. Given these findings and their association with cri-du-chat, special attention on screening examinations should include a thorough evaluation of retinal vascularization in CdC patients, even in full-term neonates.


[Strategies for the diagnosis of dementia. Experiences of collaboration between general practitioners and district nurses].
The aim was to explore the context and experiences of collaboration between the general practitioner (GP) and the district nurse (DN) on diagnosing dementia, in order to identify possible procedures to improve care. Two group interviews respectively with four DNs and five GPs working in the municipality of Copenhagen. The group interviews revealed that the suboptimised collaboration could be caused by different inter-professional diagnostic strategies and a lack of understanding of the importance of early, shared, decision-making. This could create conflicts between the groups. This study indicates a possibility of improving the collaboration between the two professional groups on diagnosing dementia. Possible approaches to improved care should focus on an inter-professional understanding of the importance of early, shared, decision-making, emphasising early identification and care of diagnosed demented patients.


Direct visualization of cytokine-producing recall antigen-specific CD4 memory T cells in healthy individuals and HIV patients.
We have used computer-assisted cytokine ELISA spot analysis to measure the frequencies, the type of cytokine, and the amount of cytokine produced by individual recall Ag-specific CD4 memory cells in freshly isolated blood. We studied the memory cells specific for tetanus toxoid and purified protein derivative in 18 healthy individuals and in 22 HIV-infected patients on highly active antiretroviral therapy (HAART). In healthy individuals, the frequency, cytokine signature, and cytokine production per cell of these memory cells were stable over time. Although it is presently unclear whether the maintenance of the memory T cell pool depends upon Ag persistence, cross-reactive Ag stimulation, or cytokine-driven bystander stimulations and expansions, our data strongly argue for a stable memory cell pool in healthy individuals. In HIV patients, however, the frequency of these memory cells was a function of the viral load. The decreased numbers of functional memory cells in patients with high viral loads might provide one mechanism behind the immunodeficient state. Although the cytokine output per cell was unaffected in most patients (20 of 24), in some patients (4 of 24) it was >100-fold reduced, which might provide an additional mechanism to account for the reduced immunocompetence of these patients. The ability to visualize directly and quantify the cytokine produced by the low frequency memory cells in freshly isolated blood that have been physiologically stimulated by Ag should aid comprehensive studies of the Ag-specific memory cell pool in vivo, in health and disease.


Structure and function of the human mitochondrial MRS2 channel.
The human Mitochondrial RNA Splicing 2 protein (MRS2) has been implicated in Mg2+ transport across mitochondrial inner membranes, thus playing an important role in Mg2+ homeostasis critical for mitochondrial integrity and function. However, the molecular mechanisms underlying its fundamental channel properties such as ion selectivity and regulation remain unclear. Here, we present structural and functional investigation of MRS2. Cryo-electron microscopy structures in various ionic conditions reveal a pentameric channel architecture and the molecular basis of ion permeation and potential regulation mechanisms. Electrophysiological analyses demonstrate that MRS2 is a Ca2+-regulated, non-selective channel permeable to Mg2+, Ca2+, Na+ and K+, which contrasts with its prokaryotic ortholog, CorA, operating as a Mg2+-gated Mg2+ channel. Moreover, a conserved arginine ring within the pore of MRS2 functions to restrict cation movements, likely preventing the channel from collapsing the proton motive force that drives mitochondrial ATP synthesis. Together, our results provide a molecular framework for further understanding MRS2 in mitochondrial function and disease.


Surgical treatment of spinal tuberculosis complicated with extensive abscess.
Tuberculosis can be responsible for extensive spinal lesions. Despite the efficacy of medical treatment, surgery is indicated to avoid or correct significant deformity, treat spinal instability, prevent neurological compromise, and to eradicate an extensive tuberculous abscess. In this paper we present our experience in the surgical management of spinal tuberculosis complicated with large abscess. Fifteen patients with spinal tuberculosis complicated with extensive abscess were identified; and nine of those patients had extension of the infection into the epidural space. The average age at treatment was 34 years old. Seven patients had thoracic infection, seven patients had lumbar infection and one had thoracolumbar infection. Six patients had neurological deficit at presentation. All patients were surgically treated with abscess debridement, spinal stabilization and concurrent antituberculous chemotherapy. A single anterior surgical approach was used in three cases, a posterior approach was used in four others and a combined approach was performed in eight patients. Surgical management allowed for effective abscess debridement and sspinal stabilization in this cohort. In combination with antituberculous drugs, surgical treatment resulted in infection eradication and bone fusion in all patients at 24 month average follow-up. Satisfactory neurological outcomes with improved American Spinal Injury Association (ASIA) scores were observed in 100% of patients. Surgical treatment for spinal tuberculosis abscess can lead to satisfactory clinical outcomes.


The DEAD-box RNA helicase RhlB is required for efficient RNA processing at low temperature in Caulobacter.
One of the most important control points in gene regulation is RNA stability, which determines the half-life of a transcript from its transcription until its degradation. Bacteria have evolved a sophisticated multi-enzymatic complex, the RNA degradosome, which is dedicated mostly to RNA turnover. The combined activity of RNase E and the other RNA degradosome enzymes provides an efficient pipeline for the complete degradation of RNAs. The DEAD-box RNA helicases are very often found in RNA degradosomes from phylogenetically distant bacteria, confirming their importance in unwinding structured RNA for subsequent degradation. This work showed that the absence of the RNA helicase RhlB in the free-living Alphaproteobacterium Caulobacter crescentus causes important changes in gene expression and cell physiology. These are probably due, at least in part, to inefficient RNA processing by the RNA degradosome, particularly at low-temperature conditions.


On the Value of Estimating Human Arm Stiffness during Virtual Teleoperation with Robotic Manipulators.
Teleoperated robotic systems are widely spreading in multiple different fields, from hazardous environments exploration to surgery. In teleoperation, users directly manipulate a master device to achieve task execution at the slave robot side; this interaction is fundamental to guarantee both system stability and task execution performance. In this work, we propose a non-disruptive method to study the arm endpoint stiffness. We evaluate how users exploit the kinetic redundancy of the arm to achieve stability and precision during the execution of different tasks with different master devices. Four users were asked to perform two planar trajectories following virtual tasks using both a serial and a parallel link master device. Users' arm kinematics and muscular activation were acquired and combined with a user-specific musculoskeletal model to estimate the joint stiffness. Using the arm kinematic Jacobian, the arm end-point stiffness was derived. The proposed non-disruptive method is capable of estimating the arm endpoint stiffness during the execution of virtual teleoperated tasks. The obtained results are in accordance with the existing literature in human motor control and show, throughout the tested trajectory, a modulation of the arm endpoint stiffness that is affected by task characteristics and hand speed and acceleration.


Localized osteomyelitis due to Mycobacterium avium complex in patients with Human Immunodeficiency Virus receiving highly active antiretroviral therapy.
We describe 3 patients who developed atypical manifestations of Mycobacterium avium complex (MAC) infection >10 months (range, 3-16 months) after attaining sustained CD4(+) T cell counts of >100 cells/microL while receiving antiretroviral therapy and not receiving MAC prophylaxis. The common features of these cases include the degree of immune reconstitution, the unusual locations of the infections, and the absence of a systemic inflammatory response. The low rate of these unusual MAC infections does not warrant continuation of primary or secondary prophylaxis after presumed immune reconstitution.


Gamma-subunits of G proteins, but not their alpha- or beta-subunits, are polyisoprenylated. Studies on post-translational modifications using in vitro translation with rabbit reticulocyte lysates.
Lipid modifications that may be introduced into several subunits of G proteins were explored by in vitro translation of recombinant mRNAs in reticulocyte lysates. In agreement with studies by others, myristic acid was incorporated into alpha i's and alpha o, but not alpha s, beta, or gamma's. In contrast, mevalonate (Mev) was incorporated only into gamma-subunits. Both, the gamma-subunit of transducin (gamma T) and that of other G proteins (gamma G) were modified by the lysates but with different characteristics. Labeled gamma T was unstable and was rapidly proteolyzed. Labeled gamma G was stable. The Mev-derivative in gamma G was sensitive to methyliodide and, after cleavage and chromatographic analysis, comigrated with the C20 polyisoprenol geranylgeraniol. This indicated that gamma G had been geranylgeranylated and that this polyisoprenoid was attached to the protein through a thioether linkage. It is thought that polyisoprenylation is defined by the COOH-terminal sequence Cys-A-A-X, where A is an aliphatic acid and X is any amino acid. Replacement by mutation of the Cys of the COOH-terminal -Cys-Ala-Ile-Leu sequence of gamma G with Ser abolished Mev incorporation, suggesting this Cys as the site of attachment of the geranylgeranyl moiety. Yet, Mev incorporation was less than 10% as much into gamma G with the Cys-A-A-X sequence -Cys-Ala-Ile-Trp. Consistent with geranylgeranylation, the C15 farnesyl moiety of farnesyl pyrophosphate was not incorporated into gamma G unless the incubations were fortified with Mev. In contrast, the farnesyl moiety was incorporated in an Mev-independent manner into gamma T (COOH terminus: -Cys-Val-Ile-Ser) and c-Ha-ras (COOH terminus: -Cys-Val-Leu-Ser) which are both farnesylated rather than geranylgeranylated. Thus, 1) separate enzymes appear to be involved in transferring farnesyl and geranylgeranyl groups to proteins, 2) structural factors other than the CAAX box contribute to the activity of the polyisoprenylating enzymes, and 3) this type of lipidation may be part of a proteolytic signaling system. Polyisoprenylation, which increases hydrophobicity of the derivatized protein, may play a role in anchoring not only ras but also G proteins to membranes.


Benign-onset acute disseminated encephalomyelitis: a report on two cases.
The signs and symptoms of acute disseminated encephalomyelitis are heterogeneous and dependent on the location and severity of the inflammatory process. The meningoencephalitic presentation may include meningism, impaired consciousness (occasionally leading to coma), seizures and confusion, or behavioral disturbances. Multifocal neurological features include a combination of optic neuritis, visual field defects, cranial neuropathy, sensorimotor impairment, ataxia, aphasia, and involuntary movements. One definition of acute disseminated encephalomyelitis is "an initial clinical event with a presumed inflammatory and demyelinating cause, with acute or sub-acute onset affecting multifocal areas of the central nervous system". Patients with acute disseminated encephalomyelitis frequently suffer from seizures, disturbances of consciousness, fever, and headaches, and occasionally there are focal signs and symptoms. Here, we report on two cases who presented with different symptoms, but the clinical findings that the patients showed were benign.


Independent regulation of Fuc-TIV and Fuc-TVII genes leading to modulation of cell surface antigen expression in developing myeloid cells.
Fucosylated antigen expression, fucosyltransferase activities and expression of Fuc-TIV and Fuc-TVII genes have been measured in the human leukemic cell lines KG1a, arrested at the undifferentiated myeloblast stage of maturation and KG1, arrested at the myeloblast and early promyelocytic stage. The results are compared with those we earlier found for the later promyelocytic cell line HL-60 and the myelocyte form into which HL-60 cells can be induced to differentiate. These leukemic cell lines, and the differentiated HL-60 cells, are believed to correspond to four successive stages of myeloid maturation in the bone marrow. Fuc-TVII mRNA was strongly expressed in the myeloblastic KG1a cells but expression was less in KG1 and HL-60 cells. In contrast to the sharp fall in Fuc-TIV expression observed on differentiation of HL-60 cells, the expression of Fuc-TIV mRNA showed a progressive increase from KG1a to HL-60 cells; thus the peak of expression of this gene was at the HL-60 promyelocyte stage. This peak correlated with an increase in fucosyltransferase activity with nonsialylated acceptors and the transitory downregulation of cell surface sialyl-Le(x) expression and upregulation of Le(x), VIM-2 and Le(y) expression. The variations in levels of expression of the fucosylated antigens on the surface of the developing myeloid cells therefore correlate with variations in mRNA expression arising from the independent regulation of Fuc-TIV and Fuc-TVII genes.


Light Deficiency Inhibits Growth by Affecting Photosynthesis Efficiency as well as JA and Ethylene Signaling in Endangered Plant Magnolia sinostellata.
The endangered plant Magnolia sinostellata largely grows in the understory of forest and suffers light deficiency stress. It is generally recognized that the interaction between plant development and growth environment is intricate; however, the underlying molecular regulatory pathways by which light deficiency induced growth inhibition remain obscure. To understand the physiological and molecular mechanisms of plant response to shading caused light deficiency, we performed photosynthesis efficiency analysis and comparative transcriptome analysis in M. sinostellata leaves, which were subjected to shading treatments of different durations. Most of the parameters relevant to the photosynthesis systems were altered as the result of light deficiency treatment, which was also confirmed by the transcriptome analysis. Gene Ontology and KEGG pathway enrichment analyses illustrated that most of differential expression genes (DEGs) were enriched in photosynthesis-related pathways. Light deficiency may have accelerated leaf abscission by impacting the photosynthesis efficiency and hormone signaling. Further, shading could repress the expression of stress responsive transcription factors and R-genes, which confer disease resistance. This study provides valuable insight into light deficiency-induced molecular regulatory pathways in M. sinostellata and offers a theoretical basis for conservation and cultivation improvements of Magnolia and other endangered woody plants.


Xanthoquinodin B3, a new anticoccidial agent produced by Humicola sp. FO-888.
Xanthoquinodin B3, a new component of anticoccidial xanthoquinodins, which was detected in the culture broth of Humicola sp. FO-888, was isolated by heat treatment of the xanthoquinodins complex. Structural elucidation indicated that xanthoquinodin B3 has the same heterodimer of xanthone- and anthraquinone-derived monomers as other xanthoquinodins. Schizont formation of monensin-resistant Eimeria tenella in BHK-21 cells was inhibited by xanthoquinodin B3 at concentrations greater than 0.035 microM.


Probing Cooperativity of N-Terminal Domain Orientations in the p97 Molecular Machine: Synergy Between NMR Spectroscopy and Cryo-EM.
The hexameric p97 enzyme plays an integral role in cellular homeostasis. Large changes to the orientation of its N-terminal domains (NTDs), corresponding to NTD-down (p97-ADP) or NTD-up (p97-ATP), accompany ATP hydrolysis. The NTDs in a series of p97 disease mutants interconvert rapidly between up and down conformations when p97 is in the ADP-bound state. While the populations of up and down NTDs can be determined from bulk measurements, information about the cooperativity of the transition between conformations is lacking. Here we use cryo-EM to determine populations of the 14 unique up/down NTD states of the homo-hexameric R95G disease-causing p97 ring, showing that NTD orientations do not depend on those of neighboring subunits. In contrast, NMR studies establish that inter-protomer cooperativity is important for regulating the orientation of NTDs in p97 particles comprising mixtures of different subunits, such as wild-type and R95G, emphasizing the synergy between cryo-EM and NMR in establishing how the components of p97 function.