Islet amyloid polypeptide: Another key molecule in Alzheimer's pathogenesis?
Recent epidemiological evidence reveals that patients suffering from type 2 diabetes mellitus (T2DM) often experience a significant decline in cognitive function, and approximately 70% of those cases eventually develop Alzheimer's disease (AD). Although several pathological processes are shared by AD and T2DM, the exact molecular mechanisms connecting these two diseases are poorly understood. Aggregation of human islet amyloid polypeptide (hIAPP), the pathological hallmark of T2DM, has also been detected in brain tissue and is associated with cognitive decline and AD development. In addition, hIAPP and amyloid β protein (Aβ) share many biophysical and physiological properties as well as exert similar cytotoxic mechanisms. Therefore, it is important to examine the possible role of hIAPP in the pathogenesis of AD. In this article, we introduce the basics on this amyloidogenic protein. More importantly, we discuss the potential mechanisms of hIAPP-induced AD development, which will be beneficial for proposing novel and feasible strategies to optimize AD prevention and/or treatment in diabetics.


The downmodulation of the foreign body reaction by cytomegalovirus encoded interleukin-10.
The foreign body reaction (FBR) is of great importance for the function and turnover of biomaterial scaffolds. The development of biological tools that modulate the FBR will augment scaffold functionality and benefit regenerative medicine. The human cytomegalovirus encodes a functional homolog of the potent anti-inflammatory human cytokine interleukin-10 (cmvIL-10). We hypothesized that cmvIL-10 downmodulates the FBR, impairing degradation of biomaterial. We studied the effect of cmvIL-10 on the FBR to subcutaneously implanted hexamethylenediisocyanate-crosslinked dermal sheep collagen (HDSC) discs in rats. CmvIL-10 impaired macrophage influx, vascularization and ingrowth into the discs up to 21 days. It also impaired the formation of giant cells and the degradation of HDSC. At day 10, deposited fibrin fibers were still present in cmvIL-10 discs. Impaired collagenase activity coincided with the impaired HDSC degradation. These results indicate that cmvIL-10 downmodulates the FBR, impairing the progression of the FBR. This study demonstrates the feasibility of interleukin-10 as a biomolecular tool in biomaterials for regenerative medicine.


Dickkopf-3 Upregulates VEGF in Cultured Human Endothelial Cells by Activating Activin Receptor-Like Kinase 1 (ALK1) Pathway.
Dkk-3 is a member of the dickkopf protein family of secreted inhibitors of the Wnt pathway, which has been shown to enhance angiogenesis. The mechanism underlying this effect is currently unknown. Here, we used cultured HUVECs to study the involvement of the TGF-β and VEGF on the angiogenic effect of Dkk-3. Addition of hrDkk-3 peptide (1 or 10 ng/ml) to HUVECs for 6 or 12 h enhanced the intracellular and extracellular VEGF protein levels, as assessed by RTPCR, immunoblotting, immunocytochemistry and ELISA. The increase in the extracellular VEGF levels was associated to the VEGFR2 activation. Pharmacological blockade of VEGFR2 abrogated Dkk-3-induced endothelial cell tubes formation, indicating that VEGF is a molecular player of the angiogenic effects of Dkk-3. Moreover, Dkk-3 enhanced Smad1/5/8 phosphorylation and recruited Smad4 to the VEGF gene promoter, suggesting that Dkk-3 activated ALK1 receptor leading to a transcriptional activation of VEGF. This mechanism was instrumental to the increased VEGF expression and endothelial cell tubes formation mediated by Dkk-3, because both effects were abolished by siRNA-mediated ALK1 knockdown. In summary, we have found that Dkk-3 activates ALK1 to stimulate VEGF production and induce angiogenesis in HUVECs.


Choice of vascular surgery as a specialty: survey of vascular surgery residents, general surgery chief residents, and medical students at hospitals with vascular surgery training programs.
Under the direction of the Association of Program Directors in Vascular Surgery, a survey was mailed to vascular surgery residents (VSRs), general surgery chief residents (GS-CRs), and fourth-year medical students (MSs) to better define reasons why trainees do and do not choose vascular surgery as a career. Questionnaires were mailed to all accredited VSR programs and their associated GS programs in the United States and Canada in 2001 (survey 1) and in 2003 (survey 2) and to 2 medical schools with VSR programs in 2001. A total of 197 VSRs, 169 GS-CRs, and 78 MSs responded (overall program response rate of 78% for VSRs, 46% for GSRs, 20% for MSs). A scoring system was assigned, with 1.0 the least important and 5.0 the most important reasons to choose or not choose vascular surgery. Technical aspects, role of mentors, and complex decision making involved in vascular surgery were the most important reasons that VSRs, GS-CRs, and MSs would choose vascular surgery as a specialty (average scores > or =4.0 for VSRs and GS-CRs; > or =3.5 for MSs). Responses of GS-CRs and VSRs did not vary significantly between surveys 1 and 2, except endovascular capabilities of vascular surgeons had a more important role in choosing vascular surgery, and future loss of patients to other interventionalists had a more important role in not choosing this specialty in the more recent survey of GS-CRs and VSRs. MSs identified lifestyle as a surgical resident (4.3) and as a surgeon (4.2) as the most important negative factors. A training paradigm consisting of 4 years general surgery + 2 years vascular surgery with a GS certificate was favored by 64% of GS-CRs and 48% of VSRs, compared with a paradigm of 5 years + 2 years with a general surgery certificate, which was favored by 29% of GS-CRs and 25% of VSRs, or 3 years + 3 years without a general surgery certificate, favored by 7% of GS-CRs and 27% of VSRs. Of note, 86% of MSs favored 3 years general surgery + 3 years vascular surgery or 2 years general surgery + 4 years vascular surgery compared with longer general surgery training periods. These findings may help vascular surgery program directors devise strategies to attract future trainees. The importance of mentorship to general surgery junior residents and medical students in choosing vascular surgery cannot be overestimated. Endovascular capabilities of vascular surgeons have an increasingly positive role in career choice by GS-CRs and VSRs, but these residents express increasing concerns about potential loss of patients to other specialists. Lifestyle concerns are the most important reasons why medical students do not choose vascular surgery as a career.


Fatal buckwheat dependent exercised-induced anaphylaxis.
Cases of food-dependent exercise-induced anaphylaxis (FEA) caused by buckwheat have been rare. Clinical, laboratory, and autopsy findings are present on an 8-year old girl with FEA caused by Japanese buckwheat. The patient consumed buckwheat noodles called "zaru soba" and immediately thereafter swam vigorously. Approximately 30 minutes later, she complained of abdominal pain, vomiting, coughing, and chest discomfort. Another ten minutes later her consciousness level deteriorated and she experienced cardiorespiratory arrest. The heart beat was restored and she was admitted to the hospital. She never regained consciousness and expired after another arrest 13 days later. Her IgE level was high (2,840 IU/ml) and the IgE-radioallergosorbent test (RAST) score was 2 for soybeans, 3 for buckwheat, 2 for rice, and 3 for wheat. An exaggerated hematemesis that occurred immediately after hospital admission indicated an inflammatory condition of the digestive tract that was caused by buckwheat. Marked ulceration accompanied with hemorrhage and necrosis was noted at the ileum. Extensive hemorrhage involving the endotracheal pulmonary field and lymphocyte infiltration of the alveolar space likely appeared after the inflammation. The analysis of buckwheat-specific IgE antibody by immunoblotting showed 7 bands that reacted with the IgE of the patient's serum, 4 bands: 16, 20, 24, and 58 kDa, were specific to the patient as compared to subjects not allergic to buckwheat. A first case of fatal FEA by buckwheat is reported with reference to specific IgE.


Manganism and idiopathic parkinsonism: similarities and differences.
From the comparison we have made between PD and manganism, we draw the following conclusions: 1. There are similarities between PD and manganism, notably the presence of (a) generalized bradykinesia and (b) widespread rigidity. 2. There are also dissimilarities between PD and manganism, notably the following in manganism: (a) less-frequent resting tremor, (b) more frequent dystonia, (c) a particular propensity to fall backward, (d) failure to achieve a sustained therapeutic response to levodopa, and (e) failure to detect a reduction in fluorodopa uptake by PET. Further studies are likely to yield more discriminants between PD and manganism. For example, PET with raclopride may be useful in early cases of manganism, and MRI may be helpful in patients with advanced manganism.


[Enhanced nisin production by overexpression of nisin immunity gene nisI in the nisin-producing strain].
Our aim was to enhance nisin production by overexpression of nisin immunity gene nisI in nisin-producing strains. Nisin immunity gene nisI with a strong promoter P59 was cloned into vector pHJ201 and introduced into Lacotococcus lactis NZ9800, resulting in a recombinant strain L. lactis NZ9800/pHMI. Then the differences between the recombinant strain and the control strain L. lactis NZ9800/pHJ201 were analyzed in several aspects, including their growth curves, nisin resistance level and antibacterial activity against indicator strain Microccus flavus NCIB 8166. The overexpression of nisI had no significant difference in growth rate between recombinant strain and contrast strain. However, it promoted recombinant strain tolerance 25% higer nisin resistance level and stronger antibacterial activity against M. flavus NCIB 8166, which was increased by 32% and 25% when fermented for 6 and 8 hours, respectively. These results indicated that overexpression of nisI gene in the nisin producing strain can effectively enhance nisin resistence level and thus improve nisin production.


Effect of Flurbiprofen and S-Flurbiprofen Patches on Multimodal Pain Management After Total Knee Arthroplasty: A Prospective Randomized Controlled Trial.
Total knee arthroplasty (TKA) is an established procedure for knee osteoarthritis. Multimodal analgesia is reportedly more effective for postoperative analgesia. We investigated the efficacy of 2 patches after TKA. Seventy-nine knees that underwent unilateral TKA for osteoarthritis were included. Oral administration, local periarticular analgesic injection, and patches were adopted for pain management. The knees were randomly assigned to the flurbiprofen patch (FPP), S-flurbiprofen patch (SFPP), and control (no patch) groups. Patch treatment was continued for 14 days. Pain according to the visual analog scale, knee flexion angle, renal dysfunction, gastrointestinal injury, duration of hospitalization, dermatitis, and the rate of using additional oral nonsteroidal anti-inflammatory drugs were compared (from preoperative to postoperative day 14). The FPP, SFPP, and control groups included 29, 27, and 23 knees, respectively. Visual analog scale was lower in the FPP and SFPP groups than in the control group on days 1 and 3 (day 1: 24.4, 25.0, and 39.4, respectively; day 3: 25.5, 23.3, and 39.3, respectively). Knee flexion angle was larger in the SFPP group than in the control group on days 7 and 14 (day 7: 89.8° and 76.6°, respectively; day 14: 98.3° and 84.2°, respectively). Neither renal dysfunction nor gastrointestinal injury was confirmed. The duration of hospitalization did not differ among the groups. Dermatitis occurred only in the SFPP group. The rate of using additional oral nonsteroidal anti-inflammatory drugs was higher in the control group. Both patches were effective and safe as part of multimodal analgesia for postoperative TKA.


Separation of a complex mixture of heparin-derived oligosaccharides using reversed-phase high-performance liquid chromatography.
A reversed-phase ion-pairing high-performance liquid chromatography (RP-HPIPC) method for the separation of a complex mixture of heparin-derived oligosacchrides has been developed by a stepwise optimization of the mobile phase, in which the concentration of ion-pairing reagent, mobile phase pH, and acetonitrile concentration were varied. The resolution of more than 30 oligosaccharide components was obtained, under optimized conditions, in an analysis time of less than 30 min. This represents the first RP-HPLC method that can separate a complex mixture of both small and large sulfated oligosaccharides in a single chromatographic step. The heparin-derived oligosaccharides, in this mixture, can also be separated under a second set of RP-HPIPC conditions using a volatile ion-pairing reagent, tributylammonium acetate, to aid in the recovery of individual sulfated oligosaccharides. Moreover, it was possible to replace sodium chloride gradient, required for eluting highly sulfated oligosaccharides, with a fixed, low concentration of a volatile salt, ammonium acetate, by utilizing an acetonitrile gradient. This solvent system might make it possible to directly interface this RP-HPIPC separation with mass spectral analysis.


Urinary steroids in young women with eating disorders.
Urinary steroid components were measured after enzyme hydrolysis and methoxym-silyl derivatization by capillary gas chromatography in young women with eating disorders. Using three internal standards, programmed temperature from 50 to 300 degrees C and flame ionization detection, on ULTRA-1 capillary column the separation of 28 steroid components is possible. Fifteen young women with different eating disorders, i.e. anorexia nervosa, bulimia nervosa and bulimarexia (aged 16-28 years) and 15 healthy women (aged 22-26 years) collected 24-h urine. A significant decrease of dehydroepiandrosterone (p<0.05) and a significant elevation of the stress marker allo-tetrahydrocorticosterone (p<0.05) were observed in the patients with compared to the controls. Among the patients two groups were defined due to a significant difference in the content of the cortisol metabolites compared to each other and to controls (p<0.01). The results confirm the role of dehydroepiandrosterone in eating disorders. The increased level of the stress marker allo-tetrahydrocorticosterone refers to the involvement of stress in these diseases, but the relevance of hormone alteration to the pathophysiology of eating disorders remains to be elucidated. The reason of the differences in cortisol metabolites levels in some patients needs further investigations.


Spinal Cord Compression By Metastatic Thoracic Spine Paraganglioma.
Paragangliomas are rare neuroendocrine tumours most commonly located in the adrenal glands. The overall incidence of paragangliomas is 0.8 per 100,000 persons, but the incidence of malignant paraganglioma was found to be 93 cases out of 400 million persons in United States. We present a case of 50 year old male who came to the hospital with back pain and progressive bilateral lower limb weakness for the past 6 months. Imaging studies revealed enhancing lesions on dorsal spines. Bone scintigraphy showed increased tracer uptake at multiple sites. Bone biopsy and immune-histochemical staining proved metastatic paraganglioma. After a thorough literature search only few cases of metastatic spine paraganglioma causing spinal cord compression have been reported to date.


The role of Zn-alpha2 glycoprotein in sperm motility is mediated by changes in cyclic AMP.
Sperm motility is essential for male reproduction or natural fertilization. The cyclic AMP (cAMP)/cAMP-dependent protein kinase A (PKA) signaling pathway is generally recognized as one of the significant signaling pathways in the regulation of mammalian spermatozoan motility. Since Zn-alpha2-glycoprotein (ZAG) activity in mammalian adipose tissue is mediated via the beta(3)-adrenoreceptor, with upregulation of the cAMP pathway, we hypothesize that ZAG may play the same role in sperm motility regulation, a new factor of regulation of sperm motility. Therefore, the gene encoding human ZAG was cloned and polyclonal antibodies were generated, and then laser scanning confocal microscopy and flow cytometry were employed to identify this protein in human spermatozoa. The results showed that ZAG protein was mostly localized on the pre-equatorial region covering the acrosome, neck, and middle piece of the flagellum of spermatozoa. Furthermore, using computer-assisted sperm analysis, we found that anti-human ZAG antibodies could significantly reduce the motility of human swim-up spermatozoa after 90- or 120-min incubation (P<0.05 and P<0.01 respectively), together with the decreasing of intracellular cAMP and PKA levels. In conclusion, these data suggest that ZAG is present in human spermatozoa and may be involved in the regulation of sperm motility via the cAMP/PKA signaling pathway.


Hospital-based diagnosis of hemorrhagic fever, encephalitis, and hepatitis in Cambodian children.
Surveillance was conducted for three clinical syndromes (hemorrhagic fever, encephalitis, and hepatitis) in Cambodian children admitted to the National Pediatric Hospital in Phnom Penh from July 1996 through September 1998. Acute- and convalescent-phase sera, and cerebrospinal fluid, when applicable, underwent diagnostic evaluation for infections with Dengue virus (DENV), Japanese encephalitis virus (JEV), and Hepatitis A, B, C, and E viruses. Of 621 children admitted with hemorrhagic fever, 499 (80%) were confirmed to have either primary or secondary DENV infection. DENV rates were as high as 10.6/100 hospital admissions in September 1998. Of 50 children with clinical encephalitis, 9 (18%) had serologic evidence of JEV infection. Forty-four children had clinical hepatitis, most (55%) due to Hepatitis A virus (HAV). One patient had Hepatitis B virus, and no patients had hepatitis C or E. This study identified a large number of children with vaccine-preventable diseases (JEV and HAV).


Predicting gas transport in formed zeolite adsorbents from NMR studies.
The self-diffusion of nitrogen, methane, and carbon monoxide within a 5A zeolitic adsorbent has been examined with use of pulsed field gradient (PFG) NMR. In all cases, the diffusion process is well-described by a refined version of the long-range diffusion model (LRDM), adapted here for use with pelletized adsorbents, which uses exclusively adsorbent porosity and isotherm data as inputs. Correlation of the experimental data with this model yields tortuosity factors that are characteristic of the adsorbate and reflect the longer diffusive path a molecule must take due to the winding nature of the pore structure. It is demonstrated that the diffusion model can be used to accurately predict the diffusion coefficients for a ternary gas mixture within a 5A zeolite. To fully characterize the diffusive process, the surface excess on the PFG NMR samples has been obtained by a novel gas-phase NMR technique that is well-suited for measuring pure and multicomponent isotherms.


Stress in residency: reality or myth?
This study was designed to determine (1) occupational stress in a group of residents in obstetrics and gynecology, (2) whether occupational stress differs between years of training, and (3) whether there is seasonal variation in occupational stress. The Occupational Stress Inventory was administered to the 44 residents in obstetrics and gynecology at our institution in September 1998. A subgroup of 24 residents were also asked to complete the occupational stress inventory in January and May 1999. T-scores of 40 to 59 are within the normal range; scores of > or =70 on the occupational role questionnaire or the psychologic strain questionnaire suggest significant maladaptive stress; scores of < or =30 on the personal resources questionnaire suggest a significant lack of coping skills. Kruskal-Wallis and repeated-measures analysis of variance were used when appropriate. Forty residents completed the occupational stress inventory in September. The median T-score for each subscale was between 40 and 60. Eleven residents (27.5%) scored > or =70 on at least 1 subscale of the occupational role questionnaire. Three residents (7.5%) scored > or =70 on at least 1 subscale of the psychologic strain questionnaire. Seven residents (17.5%) scored < or =30 on at least 1 personal resources questionnaire subscale. There was no significant difference in occupational adjustment between resident level. In the 24 residents who were followed throughout the academic year, there was no significant difference in stress at different times of the years. Occupational stress is relatively common in a group of residents in obstetrics and gynecology. There is no difference in occupational stress or personal coping skills between years of training, and there is no seasonal variation in the academic year.


Combination particles containing salmeterol xinafoate and fluticasone propionate: Formulation and aerodynamic assessment.
A precipitation process is used to produce combined particles consisting of two antiasthmatic drugs: salmeterol xinafoate and fluticasone propionate. To control the crystallisation of these particles, a micromixer is used to mix solvent and antisolvent flow. To produce particles in the respirable range, crystal growth inhibitors are added to the antisolvent flow. The obtained suspension is spray-dried afterwards to get a dry powder which can be further processed into inhalation drug products. It is found that a combination of polysorbate 80 and hydroxypropylmethyl cellulose (HPMC) represents the best excipient combination. It is supposed that the smaller molecule polysorbate rapidly stabilises the particle surface and with this inhibits crystal growth, whereas HPMC is observed to produce spherical particles during the spray-drying process acting as lubricant and dispersion-modifier. The processed particles show a needlelike habit and, therefore, tend to form aggregates. When dispersed from an inhaler device, they are only moderately disaggregated. The deposition of the single drugs salmeterol xinafoate and fluticasone propionate on the stages of the NGI shows a very uniform distribution, indicating that both drugs are evenly dispensed, with an FPF of about 22% for the combined particles which corresponds to the deposition of the marketed product.


[Investigation on the various species of bulb of fritillary and its application].
Through studies on ancient herbological works, it is claimed that the fritillary recorded in the Classic of Poet, Shennong's Herbology is not identical with the fritillary applied today. It belongs to Balbostemma paiculatum (Maxim). Frang of Cucurbitacea. Ming Yi Bie Lu is the earliest book that includes the fritillary of Lilliae as a remedy, with its species Fritillaria thunbergii mig. It can be sure that this should include the latter species. The fritillaria of Sichuan makes its appearance in Ben Cao Hui Yan of the Ming dynasty. It is derived from F. unibracteata et K. C. Hsia and F. cirrkosa D. Don.


Evidence for frozen-niche variation in a cosmopolitan parthenogenetic soil mite species (Acari, Oribatida).
Parthenogenetic lineages may colonize marginal areas of the range of related sexual species or coexist with sexual species in the same habitat. Frozen-Niche-Variation and General-Purpose-Genotype are two hypotheses suggesting that competition and interclonal selection result in parthenogenetic populations being either genetically diverse or rather homogeneous. The cosmopolitan parthenogenetic oribatid mite Oppiella nova has a broad ecological phenotype and is omnipresent in a variety of habitats. Morphological variation in body size is prominent in this species and suggests adaptation to distinct environmental conditions. We investigated genetic variance and body size of five independent forest - grassland ecotones. Forests and grasslands were inhabited by distinct genetic lineages with transitional habitats being colonized by both genetic lineages from forest and grassland. Notably, individuals of grasslands were significantly larger than individuals in forests. These differences indicate the presence of specialized genetic lineages specifically adapted to either forests or grasslands which coexist in transitional habitats. Molecular clock estimates suggest that forest and grassland lineages separated 16-6 million years ago, indicating long-term persistence of these lineages in their respective habitat. Long-term persistence, and morphological and genetic divergence imply that drift and environmental factors result in the evolution of distinct parthenogenetic lineages resembling evolution in sexual species. This suggests that parthenogenetic reproduction is not an evolutionary dead end.


First report of caprine abortions due to Chlamydia abortus in Argentina.
Infectious abortions of goats in Argentina are mainly associated with brucellosis and toxoplasmosis. In this paper, we describe an abortion outbreak in goats caused by Chlamydia abortus. Seventy out of 400 goats aborted. Placental smears stained with modified Ziehl-Neelsen stain showed many chlamydia-like bodies within trophoblasts. One stillborn fetus was necropsied and the placenta was examined. No gross lesions were seen in the fetus, but the inter-cotyledonary areas of the placenta were thickened and covered by fibrino-suppurative exudate. The most consistent microscopic finding was found in the placenta and consisted of fibrinoid necrotic vasculitis, with mixed inflammatory infiltration in the tunica media. Immunohistochemistry of the placenta was positive for Chlamydia spp. The results of polymerase chain reaction targeting 23S rRNA gene performed on placenta were positive for Chlamydia spp. An analysis of 417 amplified nucleotide sequences revealed 99% identity to those of C. abortus pm225 (GenBank AJ005617) and pm112 (GenBank AJ005613) isolates. To the best of our knowledge, this is the first report of abortion associated with C. abortus in Argentina.


Nurses' occupational physical activity levels: A systematic review.
Nurses' physical performance at work has implications both for nurses' occupational health and patient care. Although nurses are the largest healthcare workforce, are present 24-hours a day, and engage in many physically demanding tasks, nurses' occupational physical activity levels are poorly understood. The aim of this systematic review was to examine nurses' occupational physical activity levels, and explore how nurses accumulate their physical activity during a shift. This narrative systematic review was guided by the Preferred Reporting Items for Systematic Reviews and Meta-analysis (PRISMA) Statement. EBSCOHost (MEDLINE, CINAHL, Age Line, Academic Search Complete, Global Health, Health Business Centre, Health Policy Reference Centre, Health Source (Consumer and Nursing/Academic Edition) and SPORTDiscus), Embase, Informit, ProQuest Health and Medical, Science Direct, Scopus, and Web of Science databases. A systematic search of seven databases were completed to locate peer-reviewed journal articles documenting nurses' occupational physical activity levels from January 1990. Papers were included if they were original research papers; measured physical activity objectively and/or subjectively; reported nurses' occupational physical activity; and were published in English. Articles were excluded if nurses' data were not reported separately from other professional groups. Two researchers independently screened the articles, extracted data, and undertook the methodological quality assessments. Fifteen studies met the inclusion criteria. Nursing work predominantly comprised of light-intensity physical activity. In nine studies how nurses' accumulated occupational physical activity were documented and showed that the majority of a nurses' shift was spent standing or walking whilst completing direct patient care tasks. However, the definition of the nursing populations studied were often poorly reported, and few researchers reported the validity and the reliability of the measurement tools used. Nurses' occupational physical activity levels largely consist of light-intensity physical activity interspersed with moderate-intensity tasks. It is not known whether physical activity during one shift affects the activity levels in the following shift. This systematic review is the first step towards understanding the physical demands of nursing work, and how nurses' physical activity may impact workplace wellbeing and patient safety. A meta-analysis was not possible due to the variability in how physical activity outcomes were presented. Several studies had heart rate outcomes that were converted, where possible, by the authors into physical activity outcomes. This systematic review is registered with PROSPERO; Registration number: CRD42016045427.


Analysis of the ergosterol biosynthesis pathway cloning, molecular characterization and phylogeny of lanosterol 14 α-demethylase (ERG11) gene of Moniliophthora perniciosa.
The phytopathogenic fungus Moniliophthora perniciosa (Stahel) Aime & Philips-Mora, causal agent of witches' broom disease of cocoa, causes countless damage to cocoa production in Brazil. Molecular studies have attempted to identify genes that play important roles in fungal survival and virulence. In this study, sequences deposited in the M. perniciosa Genome Sequencing Project database were analyzed to identify potential biological targets. For the first time, the ergosterol biosynthetic pathway in M. perniciosa was studied and the lanosterol 14α-demethylase gene (ERG11) that encodes the main enzyme of this pathway and is a target for fungicides was cloned, characterized molecularly and its phylogeny analyzed. ERG11 genomic DNA and cDNA were characterized and sequence analysis of the ERG11 protein identified highly conserved domains typical of this enzyme, such as SRS1, SRS4, EXXR and the heme-binding region (HBR). Comparison of the protein sequences and phylogenetic analysis revealed that the M. perniciosa enzyme was most closely related to that of Coprinopsis cinerea.


Pharmacokinetic study of beta-lactam antibiotics in bronchial secretions.
The objective of the study was to evaluate the concentrations obtained in serum and bronchial secretions after administration of five beta-lactam antibiotics: cephradin (1 g per os) and cefoxitin (2 g i.v. infusion), amoxycillin (1.0 g per os), bacampicillin (0.4 g and 0.8 g per os) and ampicillin (1.0 g per os). 123 adult patients were included in the study and received a single dose of the tested drug. Serum and mucus samples were collected simultaneously 30 minutes, 1, 2 or 4 hours after administration of the drugs. Mucus samples were taken by fibroscopy but in some patients the samples were collected through a tracheostomy cannula which allowed sampling at different time intervals. The results show that the concentrations of penicillins in bronchial secretions increase progressively between one and four hours after administration of the drugs. Bronchial levels obtained after oral administration of ampicillin are low, not more than 5 to 10% of serum levels. The other antibiotics tested show worthwhile concentrations in bronchial secretions, especially with cephalosporins and bacampicillin which exhibits higher serum and bronchial concentrations than ampicillin.


Two unusual 3D honeycomb networks based on Wells-Dawson arsenomolybdates with d10 transition-metal-pyrazole connectors.
Herein, two Wells -Dawson-type arsenomolybdates, formulated as [Cu(pyr)2]6[As2Mo18O62] (1) and [Ag(pyr)2]6[As2Mo18O62] (2) (pyr = pyrazole), were hydrothermally synthesized and structurally characterized via single-crystal X-ray diffraction, elemental analysis, IR and UV-vis-NIR spectroscopies, XPS, XRD, and TG analysis. The structural analysis indicated that compounds 1 and 2 were isomorphic. They are the first reported 3D honeycomb structures of Wells-Dawson-type arsenomolybdates. The [M(pyr)2] (M = Cu and Ag) connected with [As2Mo18O62]6- polyoxoanions to form the {812·123}{8}3 topological structure. The contributions of organic ligands, pH value, reaction temperature, and transition-metal (TM) to the construction of 3D networks were elucidated. Furthermore, compounds 1 and 2 exhibited fluorescence properties in the solid state at room temperature, highly efficient catalytic ability for the degradation of five organic dyes (MB, RhB, MO, AP, and CR) under UV irradiation, and obvious electrocatalytic activities for the reduction of H2O2. The mechanisms of photocatalysis and electrocatalysis have also been discussed in detail.


A case-cohort study to investigate concomitant waterborne outbreaks of Campylobacter and gastroenteritis in Söderhamn, Sweden, 2002-3.
Increased domestic, laboratory confirmed, Campylobacter notifications were reported in Siderhamn municipality, December 2002 and January 2003. Concurrently, during preliminary investigations a large outbreak of acute gastroenteritis was detected. Simultaneously, two studies were completed to identify risk factors for infection with Campylobacter and acute gastrointestinal infection (AGI): (1) a case-cohort study using Campylobacter cases (N = 101) with a large random sample from the municipal population as referents (N = 1000) and (2) a retrospective cohort study for the outcome AGI using the same sample. A postal questionnaire was used to collect demographic, clinical, water and food consumption data. Measures of association (risk ratio (RR), odds ratio (OR)) and 95% confidence intervals (CI) were calculated. Stool, environmental and water samples were tested by standard methods at Gävle Hospital and SMI laboratories respectively. In the case-cohort study, Camplylobacter cases were more likely than referents to consume communal water (OR = 12.6 (95% CI 1.7-92.3)). In the cohort study, risk of gastroenteritis was 2.3 times higher in those who consumed water (AR = 27.3%) than others (AR = 12%). Risk of illness was associated with the amount of water consumed in both studies. Campylobacter was detected in stools and Escherichia coli (E. coli) from routine communal water (CW) samples. Results suggest both Söderhamn outbreaks of Campylobacter and AGI were associated with consumption of CW. The method used strengthened epidemiological evidence and was efficient in the use of time and resources.


Accumulation of thymol in halothane vaporizers.
Thymol may accumulate in halothane vaporizers and influence their accuracy. We determined the thymol concentration in residual liquid halothane of 28 vaporizers in regular use. Two halothane samples were brownish, probably due to a long exposure to light and irregular drainage. Irregularly drained vaporizers contained higher thymol concentrations than those drained weekly (p less than 0.05). The highest individual thymol concentration was 19 times that found in fresh halothane. Halothane vapour concentration deviated most from setting in Fluotec Mark II vaporizers, but this did not correlate with thymol concentrations in halothane liquid. The need for weekly drainage and regular service of halothane vaporizers is stressed.


Stakeholder views on the creation and use of genetically-engineered animals in research.
This interview-based study examined the diversity of views relating to the creation and use of genetically-engineered (GE) animals in biomedical science. Twenty Canadian participants (eight researchers, five research technicians and seven members of the public) took part in the interviews, in which four main themes were discussed: a) how participants felt about the genetic engineering of animals as a practice; b) governance of the creation and use of GE animals in research, and whether current guidelines are sufficient; c) the Three Rs (Replacement, Reduction, Refinement) and how they are applied during the creation and use of GE animals in research; and d) whether public opinion should play a greater role in the creation and use of GE animals. Most of the participants felt that the creation and use of GE animals for biomedical research purposes (as opposed to food purposes) is acceptable, provided that tangible human health benefits are gained. However, obstacles to Three Rs implementation were identified, and the participants agreed that more effort should be placed on engaging the public on the use of GE animals in research.


Side-Polished Fiber-Optic Line Sensor for High-Frequency Broadband Ultrasound Detection.
We demonstrate a side-polished fiber-optic ultrasound sensor (SPFS) with a broad frequency bandwidth (dc⁻46 MHz at 6-dB reduction) and a wide amplitude detection range from several kPa to 4.8 MPa. It also exhibits a high acoustic sensitivity of 426 mV/MPa with a signal-to-noise ratio of 35 dB and a noise-equivalent pressure of 6.6 kPa (over 1⁻50 MHz bandwidth) measured at 7-MHz frequency. The SPFS does not require multi-layer-coated structures that are used in other high-sensitivity optical detectors. Without any coating, this uses a microscale-roughened structure for evanescent-field interaction with an external medium acoustically modulated. Such unique structure allows significantly high sensitivity despite having a small detection area of only 0.016 mm² as a narrow line sensor with a width of 8 μm. The SPFS performance is characterized in terms of acoustic frequency, amplitude responses, and sensitivities that are compared with those of a 1-mm diameter piezoelectric hydrophone used as a reference.


Comprehensive dissection of transcript and metabolite shifts during seed germination and post-germination stages in poplar.
Seed germination, a complex, physiological-morphogenetic process, is a critical stage in the life cycle of plants. Biological changes in germinating seeds have not been investigated in poplar, a model woody plant. In this study, we exploited next-generation sequencing and metabolomics analysis and uncovered a series of significantly different genes and metabolites at various stages of seed germination and post germination. The K-means method was used to identify multiple transcription factors, including AP2/EREBP, DOF, and YABBY, involved in specific seed germination and post-germination stages. A weighted gene coexpression network analysis revealed that cell wall, amino acid metabolism, and transport-related pathways were significantly enriched during stages 3 and 5, with no significant enrichment observed in primary metabolic processes such as glycolysis and the tricarboxylic acid cycle. A metabolomics analysis detected significant changes in intermediate metabolites in these primary metabolic processes, while a targeted correlation network analysis identified the gene family members most relevant to these changing metabolites. Taken together, our results provide important insights into the molecular networks underlying poplar seed germination and post-germination processes. The targeted correlation network analysis approach developed in this study can be applied to search for key candidate genes in specific biochemical reactions and represents a new strategy for joint multiomics analyses.


[A short history of Dutch medical learning in Shizuoka Prefecture in the Edo Period].
During the Edo Period, a powerful feudal lord did not exist in Shizuoka Prefecture. In place of an influential lord, the highway "Tōkaido" controlled the district culture. Famous doctors frequently came and stayed at inns or stopped at their friends' houses. They were good friends with the general practitioners and related much news of medicine to the sons of practitioners. The sons went to Edo, Kyoto or Nagasaki and studied Dutch medicine. Their selection of masters was quite free. They put their heart into medicine. After studying, most of them returned to their homes.


[Oral rehabilitation of a patient with Wilson-syndrome using MK1 bolt attachment. A case report].
The oral condition of neurologically compromised patients seemed to be worse as compared to the healthy population, which is confirmed by concrete data in case of epilepsy patients. The dental status of our 33 years old patient with Wilson-syndrome showed the same situation. The Wilson-syndrome is a hepatolenticular degeneration, with the disfunction of copper excretion in liver. Toxic dose of copper is accumulated in liver, in the nuclei of central nerve system, in kidneys, and in cornea. Deficiency of coeruloplasmin characterizes the disease which might occur in a form of acute hepatitis, chronic active hepatitis, cirrhosis or hepatosplenomegaly without any symptom. Initial symptoms might usually be extrahepatic, with the presence of the green and golden brown Kayser-Fleischer copper deposition. Treatment is necessary until the end of life, 1 g Penicillamine/ day. Our patient with Wilson-syndrome has been treated for 17 years at the Department of Neurology Semmelweis University. Dystonia, speech disorder, and intention tremor are rampant among his symptoms. At the time of admittance to dental clinic, both the upper and lower dental arches belonged to class 2B according to the Fábián and Fejérdy classification of partial edentulousness. After root canal fillings and cementation of dowel cores, an upper and lower fix and removable partial denture combination was made. Oral hygiene was controlled every three months. Dentures are worn for half a year without complaint.


Pioglitazone protects PC12 cells against oxidative stress injury: An in vitro study of its antiapoptotic effects via the PPARγ pathway.
To the best of our knowledge, the role of peroxisome proliferator-activated receptor γ (PPARγ) in oxidative stress-induced PC12 cell damage is unknown. Using a PC12 cell model with H2O2 treatment, the present study investigated the expression levels of apoptosis-related genes and neuronal apoptosis after oxidative stress injury. The present study further investigated the protective effect and mechanism of pioglitazone, a PPARγ agonist. PC12 cells treated with H2O2 were used as a model of oxidative stress injury. An MTT assay and flow cytometry were used to detect the effect of H2O2 on PC12 cell viability and the protective effect of pioglitazone. A TUNEL assay was used to detect neuronal apoptosis. The expression levels of PPARγ, Bax, Bcl-2 and caspase-3 were examined by reverse transcription-quantitative PCR and western blotting. H2O2 reduced PC12 cell viability in a dose- and time-dependent manner. H2O2 significantly upregulated the protein expression levels of Bax and the cleaved caspase-3/caspase-3 ratio (P<0.01), decreased the protein expression levels of Bcl-2 (P<0.01), and increased the apoptosis rate of PC12 cells. Pioglitazone significantly reduced the protein expression levels of Bax and the cleaved caspase-3/caspase-3 ratio (P<0.01), increased the expression levels of Bcl-2 (P<0.01), decreased the Bax/Bcl-2 expression ratio (P<0.01) and increased the viability of H2O2-damaged PC12 cells in a dose-dependent manner. Treatment with the PPARγ antagonist GW9662 or PPARγ small interfering RNA counteracted the protective effect of pioglitazone on PC12 cells to different extents (P<0.01). Therefore, the present study reported the role of PPARγ in protecting PC12 cells against oxidative stress injury, which may lead to novel therapeutic approaches for neurodegenerative diseases.


Hepatic sarcoidosis presenting as portal hypertension and liver cirrhosis: case report and review of the literature.
Systemic sarcoidosis is a disease of unknown etiology, with the liver being the third most commonly affected organ. Most cases of hepatic sarcoidosis are not clinically apparent, but a few can progress to liver cirrhosis, portal hypertension and ultimately liver failure. The diagnosis of hepatic sarcoidosis is difficult, considering that no single laboratory test or radiographic finding can definitively diagnose this systemic disease. Diagnosis of hepatic sarcoidosis relies heavily on histopathologic evaluation of two or more organs, a diagnostic modality that is invasive and may not be applicable to all patients. The treatment of hepatic sarcoidosis is challenging, with no large randomized controlled trials done to date. Physicians must be aware of the complications of hepatic sarcoidosis, and must include the same in the differential diagnosis of liver cirrhosis. We present a case of hepatic sarcoidosis complicated by portal hypertension and liver cirrhosis.


Psychopathologic differences between cannabis-induced psychoses and recent-onset primary psychoses with abuse of cannabis.
The study aims to identify psychopathologic variables in cannabis-induced psychosis and recent-onset primary psychoses using the Symptom Checklist-90-R and the Psychiatric Research Interview for Substance and Mental Disorders. A sample of 181 subjects with psychotic symptoms and cannabis use referred to the psychiatry inpatient units of 3 university general hospitals were assessed. The final sample included 50 subjects with a diagnosis of cannabis-induced psychotic disorder (CIPD) and 104 subjects with primary psychotic disorders. Using receiver operating characteristic curves, the most efficient psychopathologic variables for classifying CIPD were interpersonal sensitivity, "depression," phobic anxiety, and Scale to Assess Unawareness of Mental Disorders subscales. The area under the receiver operating characteristic curve of the model including depression and "misattribution" scores was 96.78% (95% confidence interval, 94.43-99.13). Depressive symptoms could be used to distinguish CIPD from other primary psychotic disorders. Clinical variables related to "neurotic" symptoms could be involved in the susceptibility to cannabis-induced psychosis.


Odontocete peduncle tendons for possible control of fluke orientation and flexibility.
Dorso-ventral oscillations of cetacean caudal flukes generate lift-based thrust for swimming. Movements of the flukes are actuated by epaxial and hypaxial muscles through caudal tendons inserting onto vertebrate in the peduncle. To determine if the caudal tendons in the peduncle affect the flexibility of the flukes, we must understand how the tendons from axial muscles insert onto the caudal vertebrate. The purpose of this study was to provide a detailed description of the various tendons within the cetacean peduncle with regard to their role in swimming and flexibility. Dissection of the peduncle and flukes of multiple odontocete species showed that there were two distinct epaxial tendon sets within the peduncle: (1) extensor caudae medialis tendon (ECM) and (2) extensor caudae lateralis tendon (ECL). There is one distinct hypaxial tendon set, the medial tendon of the hypaxialis lumborum (MHL). The ECM and MHL tendons inserted serially onto caudal vertebrae while the ECL inserted exclusively onto the terminal vertebrae posterior to the fluke insertion. It is typical that tendons insert onto bone, however, the connection to the core fibrous layer of the flukes suggests an element of active control of the flexibility of the flukes via the axial muscles. Tension from muscular contraction transmitted through the tendons could affect both spanwise and chordwise flexibility. Changing flexibility could modulate thrust and efficiency over an extended operation range of swimming speeds in cetaceans.


Chinese herb Radix Polygoni Multiflori as a therapeutic drug for liver cirrhosis in mice.
Liver regeneration not only plays a functional role in directing the restoration of liver mass after resection or injury, but also may have participated in effective therapy of liver cirrhosis. Additionally, hepatocyte growth factor (HGF) appears to be a factor of great importance in liver regeneration and attenuated progression of experimental liver cirrhosis. The aim of this study is to use Radix Polygoni Multiflori (POMU) extract, a Chinese herb traditionally used for liver-protective therapy, as a reagent for the evaluation of its potential medicinal use in liver cirrhosis. We used in vitro coculture system to show that POMU could promote the expression of HGF by hepatic nonparenchymal cells, consequently the proliferation of primary liver cells and phagocytic activity of Kupffer cells using fluorescein-labeled Escherichia coli as the target, and inhibit the proliferation of stellate cells. Using dimethylnitrosamine-induced liver cirrhosis animal, POMU even at 20 mg/(kg day) dosage, was illustrated to reverse the pathogenic progression of the disease, decrease the hydroxyproline content and increases the expression of HGF messenger RNA in liver tissue. The survival rate was significantly increased in the POMU-treated animal. In conclusion, our study showed the promise of POMU in the medicinal use for the treatment of liver cirrhosis.


Knowledge and Awareness of First Aid of Avulsed Tooth among Physicians and Nurses of Hospital Emergency Department.
Physicians and nurses of emergency department are often the first line of qualified health-care providers attending the patient in person or parent's query over phone reporting immediately following the dental avulsion. Prompt action by them becomes the decisive factor in survival and prognosis of the tooth. Although a few researchers have evaluated knowledge of emergency room physicians, there is scanty of reports concerning nurses who play a climacteric role. A 10-item questionnaire regarding clinical situation apropos dental injury and tooth avulsion, inquiring knowledge of physicians and nurses was administered to 150 each of nursing professionals and physicians of the emergency department. With 100% response rate, the data obtained was subjected to statistical analysis. Results showed lack of knowledge and confidence in both groups. About 66.7% (n = 100) nurses and 74.7% (n = 112) considered reimplantation of avulsed tooth. There was overbalanced confusion regarding storage/transport media and handling of avulsed tooth. The findings from the results suggested a definite inadequacy in knowledge, demanding need for appropriate training for delivering treatment with more predictability and better prognosis.


Selecting a separable parametric spatiotemporal covariance structure for longitudinal imaging data.
Longitudinal imaging studies allow great insight into how the structure and function of a subject's internal anatomy changes over time. Unfortunately, the analysis of longitudinal imaging data is complicated by inherent spatial and temporal correlation: the temporal from the repeated measures and the spatial from the outcomes of interest being observed at multiple points in a patient's body. We propose the use of a linear model with a separable parametric spatiotemporal error structure for the analysis of repeated imaging data. The model makes use of spatial (exponential, spherical, and Matérn) and temporal (compound symmetric, autoregressive-1, Toeplitz, and unstructured) parametric correlation functions. A simulation study, inspired by a longitudinal cardiac imaging study on mitral regurgitation patients, compared different information criteria for selecting a particular separable parametric spatiotemporal correlation structure as well as the effects on types I and II error rates for inference on fixed effects when the specified model is incorrect. Information criteria were found to be highly accurate at choosing between separable parametric spatiotemporal correlation structures. Misspecification of the covariance structure was found to have the ability to inflate the type I error or have an overly conservative test size, which corresponded to decreased power. An example with clinical data is given illustrating how the covariance structure procedure can be performed in practice, as well as how covariance structure choice can change inferences about fixed effects.


Phytochemicals from Tradescantia albiflora Kunth Extracts Reduce Serum Uric Acid Levels in Oxonate-induced Rats.
Tradescantia albiflora (TA) Kunth (Commelinaceae) has been used for treating gout and hyperuricemia as folklore remedies in Taiwan. Therefore, it is worthwhile to study the effect of TA extracts on lowering uric acid activity. The hypouricemic effects of TA extracts on potassium oxonate (PO)-induced acute hyperuricemia were investigated for the first time. All treatments at the same volume (1 ml) were orally administered to the abdominal cavity of PO-induced hyperuricemic rats. One milliliter of TA extract in n-hexane (HE), ethyl acetate (EA), n-butanol (BuOH), and water fractions has 0.28, 0.21, 0.28, and 1.03 mg TA, respectively; and the plasma uric acid (PUA) level was measured for a consecutive 4 h after administration. All four fractions' extracts derived from TA were observed to significantly reduce PUA compared with the PO group. The EA-soluble fraction (TA-EA) exhibited the best xanthine oxidase (XO) inhibitory activity. Following column chromatography, 12 phytochemicals were isolated and identified from the EA fraction. The IC50 values of isolated phytochemicals indicated that bracteanolide A (AR11) showed the remarkable XO inhibitory effect (IC50 value of 76.4 μg/ml). These findings showed that the in vivo hypouricemic effect in hyperuricemic rats was consistent with in vitro XO inhibitory activity, indicating that TA extracts and derived phytochemicals could be potential candidates as hypouricemic agents. Tradescantia albiflora extracts possess in vivo hypouricemic action in hyperuricemic ratsT. albiflora extracts exhibited strong inhibitory activity against xanthine oxidase (XO)Butenolide may play an important role in XO inhibitionThe extract bracteanolide A was demonstrated potent XO inhibitory activity in vitro. Abbreviations used: TA: Tradescantia albiflora, PO: potassium oxonate, HE: n-hexane, EA: ethyl acetate, BuOH: n-butanol, PUA: plasma uric acid, XO: xanthine oxidase, MeOH: methanol, IP: intraperitoneal.


An outside-the-box activity to demonstrate how humans and animals turn.
Developing hands-on activities that engage and excite K-12 students is critical for stimulating interest in science-based careers. We created an activity for high school students that required them to integrate biology and physics concepts to experience how humans and animals maneuver through their environments (i.e., turning). Understanding how turning works is important because it accounts for up to 50% of daily walking steps and is needed for survival when animals elude predators and capture prey. For this activity, student groups used 2 × 4 lumber, wood screws, and a power drill to build an apparatus that, when connected to the body, altered rotational inertia (object's resistance to change in angular motion, I = mass × radius2). Students navigated through a slalom course with the apparatus (increased radius and rotational inertia) and without the apparatus (mass-matched control). Times to complete the course were compared between trials to determine the influence of rotational inertia on turning performance. Students compiled their data, graphed their results, and found that increased rotational inertia decreased turning performance. Results were connected to sports, rehabilitation, and dinosaur evolution. This activity was implemented during local, regional, national, and international outreach events and adapted for use in undergraduate courses as well (total impact, 250 students). At the end of the activity, students were able to 1) describe whether their results supported their hypothesis; 2) explain how radius influences rotational inertia and turning performance; and 3) apply results to real-world examples. Students and teachers appreciated this "outside-the-box" activity with an engineering twist and found it entertaining.


Immune modulation by apoptotic dental pulp stem cells in vivo.
Mesenchymal stem cells (MSCs) show considerable promise as a cellular immunotherapy for the treatment of a number of autoimmune and inflammatory disorders. However, the precise physiologically and therapeutically relevant mechanism(s) by which MSCs mediate immune modulation remains elusive. Dental pulp stem cells are a readily available source of MSCs that have been reported to show similar immune modulation in vitro as bone marrow MSCs. To test their potential in vivo, we used a clinically relevant humanized mouse model of GvHD in which only human T cells engraft. In this model, we found no effects on either T-cell proliferation, T-cell phenotype or disease progression. To determine if this lack of efficacy was related to a failure of engraftment or persistence of the cells, we used viability dependent radioactive cell tracking and showed that no cells were detectable after 24-h postinjection. Given the apparent failure of MSC to survive following intravenous injection, we hypothesized that their apoptosis may account for the widely reported therapeutic effect in numerous experimental models in vivo. To address this, we employed a well-established model of allergic airway inflammation to compare the efficacy of live and apoptotic MSCs in a fully immunocompetent model. In this model, both live and apoptotic dental pulp MSCs induced a robust immune suppressive reaction that was substantially greater with apoptotic cells. We propose that the mechanism of immune modulation following systemic application of MSCs is a result of cell entrapment and apoptosis occurring in the lungs.


Histological study of granulated metrial gland (GMG) cells in beige (DA-bg/bg) rats.
To clarify the roles of granulated metrial gland (GMG) cells for successful pregnancy in rats, GMG cells in beige rats (genotype: DA-bg/bg), whose NK cells show lysosomal dysfunction because of abnormalities in cytoplasmic granules, were examined in mid- and late-pregnancy by light and electron microscopies. The GMG cells of beige rats were significantly less in number than those of the two controls (genotypes: DA-bg/+ and DA-+/+) in mid- and late-pregnancy, and this accompanied a low reproductive performance in the beige rats. The size of intracellular granules in the GMG cells of the beige rats was larger than for the two controls on each corresponding day of pregnancy. These results suggest that the activity of rat GMG cells and peripheral NK cells might be influenced by the beige gene, which is involved in reproductive performance.


Effects of direct adsorption of lipoproteins apheresis on lipoproteins, low-density lipoprotein subtypes, and hemorheology in hypercholesterolemic patients with coronary artery disease.
Direct adsorption of lipoproteins (DALI) apheresis has been shown to reduce effectively low-density lipoprotein (LDL) cholesterol and lipoprotein (a) concentrations. However, the effects on nontraditional risk indicators such as hemorheology and LDL subtypes have not been investigated so far. Five patients (2 women, 3 men, age 53 +/- 8 years) with coronary artery disease and severe LDL hypercholesterolemia regularly treated with other LDL apheresis devices entered the study and were then treated with DALI for the first time. Hemorheological and lipoprotein parameters were measured before and immediately after the initial DALI apheresis as well as before the fourth DALI apheresis. Compared to baseline (before the first DALI apheresis), the following parameters were significantly improved (p < 0.05) after the first DALI apheresis: LDL cholesterol (69 +/- 28 versus 208 +/- 82 mg/dl) and cholesterol in each LDL subfraction as well as plasma viscosity (1.23 +/- 0.04 versus 1.37 +/- 0.06 mPa), C-reactive protein, native blood viscosity, red cell aggregation, and red cell deformability. When parameters before the fourth DALI apheresis were compared to baseline, LDL cholesterol was still lower, and red cell deformability was still improved while cholesterol in each subfraction showed a statistical trend to lower concentrations (0.08 < p < 0.14). In conclusion, DALI apheresis not only reduces LDL cholesterol but also induced a significant reduction of cholesterol in all LDL subfractions and improved various hemorheological parameters.


Reactive proliferative lesions in lymph nodes from rheumatoid arthritis patients. A clinicopathological and immunohistological study.
In 22 cases of rheumatoid arthritis (RA), including 4 cases of malignant RA (MRA), reactive proliferative lymph node lesions were studied clinicopathologically and immunohistochemically. This series included 5 males and 17 females. The period between disease onset and lymph node biopsy ranged from 3 months to 41 years. Generalized lymphadenopathy was noted in 13 cases and constitutional symptoms in 8. The histological findings characteristic of RA were 1) follicular hyperplasia with active germinal centers and 2) polyclonal plasma cell infiltration in the interfollicular area. Studies of intracytoplasmic immunoglobulin showed that gamma-heavy chain-expressing plasma cells were a major component in the interfollicular area in 17 RA cases. However, in 4 MRA cases, a prominent increase of mu chain-expressing plasma cells was recognized in the same area. In the 3 cases for which fresh tissue sections were stained with monoclonal antibodies against lymphocytes, we found that the majority of T cells in the interfollicular area had helper/inducer markers. The identical locations of the T cell population and plasma cells indicated that both played a role in the proliferation and/or differentiation of B cells in lymph nodes in RA.


EBV-associated hemophagocytic lymphohistiocytosis complicated by severe coagulation disorders and opportunistic infections: case report of a survivor.
The possibility of hemophagocytic lymphohistiocytosis should always be kept in mind when examining/treating a patient with fever of unknown origin and sepsis-like symptoms. Early diagnosis leading to prompt initiation of immunosuppressive therapy as well as aggressive supportive care, including correction of coagulation abnormalities and treatment of opportunistic infections, can decrease mortality.


Driving Cancer Tumorigenesis and Metastasis Through UPR Signaling.
In the tumor microenvironment, cancer cells encounter both external and internal factors that can lead to the accumulation of improperly folded proteins in the Endoplasmic Reticulum (ER) lumen, thus causing ER stress. When this happens, an adaptive mechanism named the Unfolded Protein Response (UPR) is triggered to help the cell cope with this change and restore protein homeostasis in the ER. Sequentially, one would expect that the activation of the three UPR branches, driven namely by IRE1, PERK, and ATF6, are crucial for the adaptation of cancer cells to the changing environment and thus for their survival and further propagation. Indeed, in the last few years, an increasing amount of studies has shown the implication of UPR signaling in different aspects of carcinogenesis and tumor progression. Features such as sustaining proliferation and resistance to cell death, genomic instability, altered metabolism, increased inflammation and tumor-immune infiltration, invasion and metastasis, and angiogenesis, defined as "the hallmarks of cancer", can be regulated by the UPR machinery. At the same time, new potential therapeutic interventions applicable to different kinds of cancers are being revealed. In order to describe the emerging role of UPR in cancer biology, these are the points that will be discussed in this chapter.


[Statins and muscular side-effects].
Statins are effective in the treatment of hypercholesterolemia for primary and secondary prevention of cardiovascular disease. While most side effects of statins are mild and transient, muscular symptoms are relatively common (5 to 10% of patients), but rarely serious (myositis, rhabdomyolysis). In cases of myopathy, the severity of symptoms and the determination of CK (creatine kinase) determine whether discontinuation of statin is necessary. Alternative strategies are also suggested. This article reviews suggestions on the management of these complaints that are a challenge in clinical practice.


A twin‑pair analysis indicates congenital scoliosis is associated with allele‑specific methylation in the SVIL gene.
Congenital scoliosis (CS) is a congenital disease resulting in abnormal vertebral development. Several studies have indicated that both genetic and environmental factors during pregnancy increase the risk of CS development. However, the exact mechanisms underlying CS pathogenesis remain unknown. To address this issue, both genetic (by whole‑exome sequencing) and epigenetic (by methylated DNA immunoprecipitation sequencing) maps from CS disease‑discordant monozygotic twins were generated in the present study. The differences in the presence of common and rare single nucleotide polymorphisms and in methylation patterns between the twins were investigated. The results indicated that rare mutations were more likely to underlie CS development compared with common mutations. Furthermore, differences in the allele‑specific methylation pattern in the supervillin (SVIL) gene between the twins were identified. It has been reported that SVIL exerts a number of functions associated with CS, indicating its role as a novel mechanism promoting CS pathogenesis.


Genetic insight into the putative causal proteins and druggable targets of osteoporosis: a large-scale proteome-wide mendelian randomization study.
Background: Osteoporosis is a major causative factor of the global burden of disease and disability, characterized by low bone mineral density (BMD) and high risks of fracture. We aimed to identify putative causal proteins and druggable targets of osteoporosis. Methods: This study utilized the largest GWAS summary statistics on plasma proteins and estimated heel BMD (eBMD) to identify causal proteins of osteoporosis by mendelian randomization (MR) analysis. Different GWAS datasets were used to validate the results. Multiple sensitivity analyses were conducted to evaluate the robustness of primary MR findings. We have also performed an enrichment analysis for the identified causal proteins and evaluated their druggability. Results: After Bonferroni correction, 67 proteins were identified to be causally associated with estimated BMD (eBMD) (p < 4 × 10-5). We further replicated 38 of the 67 proteins to be associated with total body BMD, lumbar spine BMD, femoral neck BMD as well as fractures, such as RSPO3, IDUA, SMOC2, and LRP4. The findings were supported by sensitivity analyses. Enrichment analysis identified multiple Gene Ontology items, including collagen-containing extracellular matrix (GO:0062023, p = 1.6 × 10-10), collagen binding (GO:0005518, p = 8.6 × 10-5), and extracellular matrix structural constituent (GO:0005201, p = 2.7 × 10-5). Conclusion: The study identified novel putative causal proteins for osteoporosis which may serve as potential early screening biomarkers and druggable targets. Furthermore, the role of plasma proteins involved in collagen binding and extracellular matrix in the development of osteoporosis was highlighted. Further studies are warranted to validate our findings and investigate the underlying mechanism.


The Chicago Water Isotope Spectrometer (ChiWIS-lab): A tunable diode laser spectrometer for chamber-based measurements of water vapor isotopic evolution during cirrus formation.
We describe a new tunable diode laser (TDL) absorption instrument, the Chicago Water Isotope Spectrometer, designed for measurements of vapor-phase water isotopologues in conditions characteristic of the upper troposphere [190-235 K temperature and 2-500 parts per million volume (ppmv) water vapor]. The instrument is primarily targeted for measuring the evolving ratio of HDO/H2O during experiments in the "Aerosol Interaction and Dynamics in the Atmosphere" (AIDA) cloud chamber. The spectrometer scans absorption lines of both H2O and HDO near the 2.64 µm wavelength in a single current sweep, increasing the accuracy of isotopic ratio measurements. At AIDA, the instrument is configured with a 256-m path length White cell for in situ measurements, and effective sensitivity can be augmented by enhancing the HDO content of chamber water vapor by an order of magnitude. The instrument has participated to date in the 2012-2013 IsoCloud campaigns studying isotopic partitioning during the formation of cirrus clouds and in the AquaVIT-II instrument intercomparison campaign. Realized precisions for 1-s measurements during these campaigns were 22 ppbv for H2O and 16 ppbv for HDO, equivalent to relative precisions of less than 0.5% for each species at 8 ppmv water vapor. The 1-s precision of the [HDO]/[H2O] ratio measurement ranged from 1.6‰ to 5.6‰ over the range of experimental conditions. H2O measurements showed agreement with calculated saturation vapor pressure to within 1% in conditions of sublimating ice and agreement with other AIDA instruments (the AIDA SP-APicT reference TDL instrument and an MBW 373LX chilled mirror hygrometer) to within 2.5% and 3.8%, respectively, over conditions suitable for all instruments (temperatures from 204 K to 234 K and H2O content equivalent to 15-700 ppmv at 200 hPa).


Development of Research into Autophagic Lysosome Reformation.
Autophagy is a lysosome-dependent degradation process that is essential for maintaining cellular homeostasis. In recent years, more studies have focused on the late stages of autophagy. Our group discovered and studied the terminal step of autophagy, namely autophagic lysosome reformation (ALR). ALR is the process that regenerates functional lysosomes from autolysosomes, thus maintaining lysosome homeostasis. ALR involves clathrin-mediated membrane budding from autolysosomes, elongation of membrane tubules along microtubules with the pulling force provided by the motor protein KIF5B, proto-lysosome scission by dynamin 2, and finally maturation of proto-lysosomes to functional lysosomes. In this review, we will summarize progress in unveiling the molecular mechanisms underlying ALR and its potential pathophysiological roles.


Spontaneous wound dehiscence after removal of single continuous penetrating keratoplasty suture: conservative management.
To describe a case of spontaneous wound dehiscence (WD) following removal of single continuous penetrating keratoplasty (PKP) suture, who was treated conservatively with a bandage contact lens. A 36 year-old man who had penetrating keratoplasty for keratoconus 15 months earlier and removal of the single continues suture the previous week underwent ocular examination. He was treated with topical dexamethasone 0.3%, chloramphenicol 0.5% and cyclopentolate 1% 3 times a day and had a bandage contact lens (BCL) inserted. On presentation, he was complaining of reduced vision, tearing and pain following an attack of rhinitis-induced sneezing. His visual acuity (VA) in the affected eye was counting fingers, the anterior chamber was formed, the intraocular pressure (IOP) was low and there was a 2-clock-hour WD with a positive Seidel test but no iris incarceration. Two months later, his corrected VA was 6/5, his IOP was normal and the graft-host junction was good with no uplift. A 2-clock-hour WD after single continuous PKP suture removal may be conservatively treated with a BCL.


Comparative Proteomics of Rat Olfactory Bulb Reveal Insights into Susceptibility and Resiliency to Chronic-stress-induced Depression or Anxiety.
Chronic stress causes the abnormality of olfactory bulb (OB) in both anxiety and depression, however, the unique and common neurobiological underpinnings are still poorly understood. Previously, we built the three groups by chronic mild stress (CMS), depression-susceptible (Dep-Sus): with depression-like behavior, anxiety-susceptible (Anx-Sus): with anxiety-like behavior and insusceptible (Insus): without depression- and anxiety-like behaviors. To continuously explore the protein expression changes in these three groups, comparative quantitative proteomics analysis was conducted on the rat OB as crucial part of the olfactory system. Next, bioinformatics analyses were implemented whereas protein expressions were independently analyzed by parallel reaction monitoring (PRM) or Western blot (WB). The OB-proteome analysis identified totally 133 differentially expressed proteins as a CMS response. These deregulated proteins were involved in multiple functions and significant pathways potentially correlated with phenotypes of maladaptive behavior of depression or anxiety as well as adaptive behavior, and hence might act as potential candidate protein targets. The subsequent PRM-based or WB-based analyses showed that changes in Nefl, Mtmr7 and Tk2; Prkaca, Coa3, Cox6c2, Lamc1 and Tubal3; and Pabpn1, Nme3, Sos1 and Lum were uniquely associated with Dep-Sus, Anx-Sus, and Insus groups, respectively. These phenotype-specific deregulated proteins were primarily involved in multiple metabolic and signaling pathways, suggesting that the identical CMS differently impacted the olfactory protein regulation system and biological processes. To sum up, our present data as a useful proteomics underpinning provided the common and distinct molecular insights into the biochemical understanding of OB dysfunction underlying susceptibility and resiliency to chronic-stress-induced anxiety or depression.


CAPTURE-JIA: a consensus-derived core dataset to improve clinical care for children and young people with juvenile idiopathic arthritis.
Data collected during routine clinic visits are key to driving successful quality improvement in clinical services and enabling integration of research into routine care. The purpose of this study was to develop a standardized core dataset for juvenile idiopathic arthritis (JIA) (termed CAPTURE-JIA), enabling routine clinical collection of research-quality patient data useful to all relevant stakeholder groups (clinicians, service-providers, researchers, health service planners and patients/families) and including outcomes of relevance to patients/families. Collaborative consensus-based approaches (including Delphi and World Café methodologies) were employed. The study was divided into discrete phases, including collaborative working with other groups developing relevant core datasets and a two-stage Delphi process, with the aim of rationalizing the initially long data item list to a clinically feasible size. The initial stage of the process identified collection of 297 discrete data items by one or more of fifteen NHS paediatric rheumatology centres. Following the two-stage Delphi process, culminating in a consensus workshop (May 2015), the final approved CAPTURE-JIA dataset consists of 62 discrete and defined clinical data items including novel JIA-specific patient-reported outcome and experience measures. CAPTURE-JIA is the first 'JIA core dataset' to include data items considered essential by key stakeholder groups engaged with leading and improving the clinical care of children and young people with JIA. Collecting essential patient information in a standard way is a major step towards improving the quality and consistency of clinical services, facilitating collaborative and effective working, benchmarking clinical services against quality indicators and aligning treatment strategies and clinical research opportunities.


Carotid angioplasty and stenting. Will they match the gold standard?
Technological advances in endoluminal equipment have changed treatment strategies for vascular disease. While the successful results of intervention in the subclavian and innominate arteries are well documented, our experience in the carotid arterial region is still in the early stages, and we are mindful of the potential for neurologic complications. We now find ourselves in the position of comparing the results of endovascular intervention with those of what has become a classic, "gold standard" procedure, carotid endarterectomy. Although we have yet to determine definitively the superiority of one method over another, it has become clear that some carotid lesions are considerably more amenable to endovascular treatment than others. We must evaluate the type and location of lesions, as well as the technical features of carotid angioplasty and stent placement, when we compare the results of endovascular and open procedures.


Ethylene Response Factor070 regulates root development and phosphate starvation-mediated responses.
Inorganic phosphate (Pi) availability is a major factor determining growth and consequently the productivity of crops. However, it is one of the least available macronutrients due to its high fixation in the rhizospheres. To overcome this constraint, plants have developed adaptive responses to better acquire, utilize, and recycle Pi. Molecular determinants of these adaptive mechanisms include transcription factors (TFs) that play a major role in transcriptional control, thereby regulating genome-scale networks. In this study, we have characterized the biological role of Arabidopsis thaliana Ethylene Response Factor070 (AtERF070), a Pi starvation-induced TF belonging to the Apetala2/Ethylene Response Factor family of TFs in Arabidopsis (Arabidopsis thaliana). It is localized to the nucleus and induced specifically in Pi-deprived roots and shoots. RNA interference-mediated suppression of AtERF070 led to augmented lateral root development resulting in higher Pi accumulation, whereas there were reductions in both primary root length and lateral root number in 12-d-old transgenic seedlings overexpressing AtERF070. When the overexpressing lines were grown to maturity under greenhouse conditions, they revealed a stunted bushy appearance that could be rescued by gibberellic acid application. Furthermore, a number of Pi starvation-responsive genes were modulated in AtERF070-overexpressing and RNA interference lines, thereby suggesting a potential role for this TF in maintaining Pi homeostasis.


Ascomylactams A-C, Cytotoxic 12- or 13-Membered-Ring Macrocyclic Alkaloids Isolated from the Mangrove Endophytic Fungus Didymella sp. CYSK-4, and Structure Revisions of Phomapyrrolidones A and C.
Three new 12- or 13-membered-ring macrocyclic alkaloids, named ascomylactams A-C (1-3), along with the analogues phomapyrrolidone C (4) and phomapyrrolidone A (5) were isolated from the mangrove endophytic fungus Didymella sp. CYSK-4. Their structures were elucidated by analysis of extensive spectroscopic data and mass spectrometric data. The structures and absolute configurations of 1 and 2 were determined by single-crystal X-ray diffraction experiments, which represents the first crystal structures described for a (6/5/6/5) tetracyclic skeleton fused with a 12- or 13-membered-ring macrocyclic moiety. The configurations of phomapyrrolidone C (4) and phomapyrrolidone A (5) were revised by detailed analysis of the NMR data. In a cytotoxic assay, compounds 1 and 3 showed moderate cytotoxicity against MDA-MB-435, MDA-MB-231, SNB19, HCT116, NCI-H460, and PC-3 human cancer cell lines, with IC50 values in the range of 4.2-7.8 μM.


Tau protein modulates an epigenetic mechanism of cellular senescence in human SH-SY5Y neuroblastoma cells.
Introduction: Progressive Tau deposition in neurofibrillary tangles and neuropil threads is the hallmark of tauopathies, a disorder group that includes Alzheimer's disease. Since Tau is a microtubule-associated protein, a prevalent concept to explain the pathogenesis of tauopathies is that abnormal Tau modification contributes to dissociation from microtubules, assembly into multimeric β-sheets, proteotoxicity, neuronal dysfunction and cell loss. Tau also localizes in the cell nucleus and evidence supports an emerging function of Tau in DNA stability and epigenetic modulation. Methods: To better characterize the possible role of Tau in regulation of chromatin compaction and subsequent gene expression, we performed a bioinformatics analysis of transcriptome data obtained from Tau-depleted human neuroblastoma cells. Results: Among the transcripts deregulated in a Tau-dependent manner, we found an enrichment of target genes for the polycomb repressive complex 2. We further describe decreased cellular amounts of the core components of the polycomb repressive complex 2 and lower histone 3 trimethylation in Tau deficient cells. Among the de-repressed polycomb repressive complex 2 target gene products, IGFBP3 protein was found to be linked to increased senescence induction in Tau-deficient cells. Discussion: Our findings propose a mechanism for Tau-dependent epigenetic modulation of cell senescence, a key event in pathologic aging.


Popliteal angle in infants with west syndrome.
The aim of this study was to clarify the relationship between neurologic findings and outcome of patients with West syndrome, focusing on the popliteal angle. The complete neurologic examination, including an assessment of the popliteal angle and muscle tone, was performed on 45 patients with West syndrome. A tight popliteal angle was determined when it was 120 degrees or less. In all 45 patients, abnormal muscle tone was not correlated with any variables. A tight popliteal angle was correlated with seizure persistence, cerebral palsy, and abnormality on magnetic resonance imaging (MRI), as well as severe developmental delay. When limited to the patients with developmental delay, a tight popliteal angle was correlated with severe developmental delay, cerebral palsy, and MRI abnormality, although abnormal muscle tone was not correlated with any items. In the delay group, eight patients had a tight popliteal angle with normal muscle tone. Among them, severe developmental delay was seen in seven (88%), seizure persistence in five (63%), and MRI abnormality in five (63%). These results suggest that a tight popliteal angle might be an indicator of poor neurologic outcome in patients with West syndrome.


Quantification of hydrogen cyanide (HCN) in breath using selected ion flow tube mass spectrometry--HCN is not a biomarker of Pseudomonas in chronic suppurative lung disease.
Hydrogen cyanide (HCN) in exhaled breath has been proposed as a biomarker for airway inflammation, and also a marker of the presence in the airways of specific organisms, especially Pseudomonas aeruginosa. However the production of HCN by salivary peroxidase in the oral cavity increases orally exhaled concentrations, and may not reflect the condition of the lower airways. Using SIFT-MS we aimed to determine an appropriate single-exhalation breathing maneuver which avoids the interference of HCN produced in the oral cavity. We have established that the SIFT-MS Voice200™ is suitable for the online measurement of HCN in exhaled breath. In healthy volunteers a significantly higher end exhaled HCN concentration was measured in oral exhalations compared to nasal exhalations (mean ± SD) 4.5 ± 0.6 ppb versus 2.4 ± 0.3 ppb, p < 0.01. For the accurate and reproducible quantification of end exhaled HCN in breath a nasal inhalation to full vital capacity and nasal exhalation at controlled flow is recommended. This technique was subsequently used to measure exhaled HCN in a group of patients with chronic suppurative lung disease (CSLD) and known microbiological colonization status to determine utility of HCN measurement to detect P. aeruginosa. Median nasal end exhaled HCN concentrations were higher in patients with CSLD (3.7 ppb) than normal subjects (2.0 ppb). However no differences between exhaled HCN concentrations of subjects colonized with P. aeruginosa and other organisms were identified, indicating that breath HCN is not a suitable biomarker of P. aeruginosa colonization.


Elevated Serum Regulator of Calcineurin 2 is Associated With an Increased Risk of Non-Alcoholic Fatty Liver Disease.
Background: The promoting effect of the regulator of calcineurin 2 (RCAN2) in hepatic steatosis has been observed in animal studies. However, the association of RCAN2 with non-alcoholic fatty liver disease (NAFLD) in humans remains unclear. This study aimed to evaluate the expression of RCAN2 in the liver of mice with hepatic steatosis and in the serum of NAFLD patients and to explore the relationship between serum RCAN2 levels and NAFLD. Methods: The mRNA and protein expression of RCAN2 were detected by quantitative real-time PCR (qRT-PCR) and Western blot. NAFLD was diagnosed by abdominal ultrasonography. Circulating RCAN2 levels were measured by ELISA kits. The relationship between serum RCAN2 levels and NAFLD was assessed. Results: qRT-PCR and Western blot analysis showed that compared with the corresponding controls, the mRNA and protein expression of RCAN2 were significantly increased in the liver tissues of db/db and mice on a high-fat diet. Serum RCAN2 levels were markedly elevated in NAFLD patients compared with non-NAFLD subjects. Binary logistic regression analysis showed that serum RCAN2 levels were significantly associated with NAFLD. Receiver operation characteristic (ROC) curve analysis showed that serum RCAN2 might act as a predictive biomarker for NAFLD [area under the curve (AUC) = 0.663, 95% CI = 0.623-0.702], and the serum RCAN2/(AST/ALT) ratio displayed improved predictive accuracy (AUC = 0.816, 95% CI = 0.785-0.846). Conclusion: Elevated serum RCAN2 levels were associated with an increased risk of NAFLD. Serum RCAN2, especially the serum RCAN2/(AST/ALT) ratio, might be a candidate diagnostic marker for NAFLD.


Stability and Instrumentation Stresses Among Sacropelvic Fixation Techniques With Novel Porous Fusion/Fixation Implants: A Finite Element Study.
Sacropelvic fixation is frequently combined with thoracolumbar instrumentation for correcting spinal deformities. This study aimed to characterize sacropelvic fixation techniques using novel porous fusion/fixation implants (PFFI). Three T10-pelvis finite element models were created: (1) pedicle screws and rods in T10-S1, PFFI bilaterally in S2 alar-iliac (S2AI) trajectory; (2) fixation in T10-S1, PFFI bilaterally in S2AI trajectory, triangular implants bilaterally above the PFFI in a sacro-alar-iliac trajectory (PFFI-IFSAI); and (3) fixation in T10-S1, PFFI bilaterally in S2AI trajectory, PFFI in sacro-alar-iliac trajectory stacked cephalad to those in S2AI position (2-PFFI). Models were loaded with pure moments of 7.5 Nm in flexion-extension, lateral bending, and axial rotation. Outputs were compared against 2 baseline models: (1) pedicle screws and rods in T10-S1 (PED), and (2) pedicle screws and rods in T10-S1, and S2AI screws. PFFI and S2AI resulted in similar L5-S1 motion; adding another PFFI per side (2-PFFI) further reduced this motion. Sacroiliac joint (SIJ) motion was also similar between PFFI and S2AI; PFFI-IFSAI and 2-PFFI demonstrated a further reduction in SIJ motion. Additionally, PFFI reduced max stresses on S1 pedicle screws and on implants in the S2AI position. The study shows that supplementing a long construct with PFFI increases the stability of the L5-S1 and SIJ and reduces stresses on the S1 pedicle screws and implants in the S2AI position. The findings suggest a reduced risk of pseudarthrosis at L5-S1 and screw breakage. Clinical studies may be performed to demonstrate applicability to patient outcomes. Not applicable (basic science study).


Altered neuronal nitric oxide synthase in the aging vascular system: implications for estrogens therapy.
Ovarian dysfunction at any age is associated with increased cardiovascular risk in women; however, therapeutic effects of exogenous estrogens are age dependent. Estradiol (E2) activates neuronal nitric oxide synthase (nNOS) in vascular cells. Because nNOS is prone to uncoupling under unfavorable biochemical conditions (as seen in aging), E2 stimulation of nNOS may lack vascular benefits in aging. Small mesenteric arteries were isolated from female Sprague Dawley rats, 3 or 12 months old, who were ovariectomized (Ovx) and treated with placebo or E2 for 4 wk. Vascular relaxation to exogenous E2 (0.001-100 μmol/liter) ± selective nNOS inhibitor (N-propyl-l-arginine, 2 μmol/liter) or pan-NOS inhibitor [Nω-nitro-l-arginine methyl ester (l-NAME), 100 μmol/liter] was examined on wire myograph. NOS expression was measured by Western blotting in thoracic aortas, in which superoxide generation was detected as dihydroethidium (DHE) fluorescence. E2 relaxations were impaired in Ovx conditions. E2 treatment (4 wk) normalized vascular function in young rats only. Both l-N-propyl-l-arginine and l-NAME blunted E2 relaxation in young controls, but only l-NAME did so in aging controls. NOS inhibition had no effect on acute E2 relaxation in Ovx rats, regardless of age or treatment. nNOS expression was similar in all animal groups. However, nNOS inhibition increased DHE fluorescence in young controls, whereas it reduced it in aging or Ovx animals. In E2-treated animals of either age, superoxide production was NOS independent. In conclusion, nNOS contributed to vascular relaxation in young, but not aging rats, where its enzymatic function shifted toward superoxide production. Thus, nNOS dysfunction may explain a mechanism of impaired E2 signaling in aging conditions.


[Shoe factory workers, solvents and health].
Exposure to organic solvents in footwear manufacturing industry came from the glues used adhering the shoe parts to each other. Benzene was the first solvent used in shoe factories until the evidence of its capacity to cause leukaemia. Then, the demonstration that exposure to n-hexane was related to distal polyneuropathy limited the use of this substance. After that, results of neurotoxicological studies conducted on workers exposed to different mixtures of organic solvents make necessary prevention measure directed to a progressive reduction of air dispersion of these chemicals. Today exposure to solvents in workplaces is regulated by health based exposure limit values that should warranty absence of central nervous system effects. One of the most important rules of occupational medicine is verify that these exposure levels are really health protective also for workers with increased susceptibility.


Total adiponectin and risk of symptomatic lower extremity peripheral artery disease in men.
Lower concentrations of adiponectin have been linked to subsequent risk of coronary heart disease in healthy individuals. Whether similar relationships exist for the development of systemic atherosclerosis, such as peripheral artery disease (PAD), is uncertain. We investigated the association between total adiponectin and risk of lower extremity PAD. We performed a prospective, nested case-control study among 18,225 male participants of the Health Professionals Follow-up Study who were free of diagnosed cardiovascular disease at the time of blood draw (1993-1995). During 14 years of follow-up, 143 men developed PAD. Using risk set sampling, controls were selected in a 3:1 ratio and matched on age, smoking status, fasting status, and date of blood draw (n=429). Median (interquartile range) adiponectin concentrations at baseline were lower among cases compared with controls (4.1 [3.2-5.5] versus 5.4 [3.8-7.5] µg/mL; P<0.001). A log-linear inverse association was evident over the full spectrum of adiponectin concentrations with PAD risk after controlling for baseline cardiovascular risk factors using restricted spline conditional logistic regression. Adiponectin was associated with a 42% lower risk of PAD per SD increase in natural log-transformed adiponectin (relative risk, 0.58; 95% confidence interval, 0.45-0.74) after adjustment for cardiovascular risk factors. The relative risk was attenuated (relative risk, 0.68; 95% confidence interval, 0.51-0.92) after further accounting for high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, C-reactive protein, and cystatin C. Additional adjustment for hemoglobin A(1c), triglycerides, and γ-glutamyltransferase had little impact on this association (relative risk, 0.68; 95% confidence interval, 0.50-0.92). Total adiponectin is inversely associated with risk of symptomatic lower extremity PAD in men.


Creating Complex Fluorophore Spectra on Antibodies Through Combinatorial Labeling.
Fluorescently-labeled antibodies are central to many biochemical assays, but they are not easy to multiplex beyond 3-4 colors. A long-term hypothesis of ours is that labeling antibodies with multiple fluorophores, in a way such that fluorescence resonance energy transfer (FRET) occurs, may provide a way to increase fluorescence multiplexing ability by creating a rich variety of complex emission spectra that could be deconvolved via spectral methods. However, it is not yet clear how one can effectively label antibodies with multiple fluorophores that exhibit FRET. Here, we show how to use Mix-n-Stain antibody labeling kits from Biotium to label antibodies with multiple fluorophores that exhibit FRET. Key to our approach is the use of Fab fragments, as opposed to full IgG molecules, since the full IgG molecules are generally too large to allow the fluorophore proximity necessary for observable FRET. We show that our approach works with two different sets of FRET-capable fluorophore combinations: CF405M/CF488A and CF568/CF640R. These results form the basis for continued development of approaches for increased multiplexing of fluorescent antibody measurements.


A single discrete Rab5-binding site in phosphoinositide 3-kinase β is required for tumor cell invasion.
Phosphoinositide 3-kinase β (PI3Kβ) is regulated by receptor tyrosine kinases (RTKs), G protein-coupled receptors (GPCRs), and small GTPases such as Rac1 and Rab5. Our lab previously identified two residues (Gln596 and Ile597) in the helical domain of the catalytic subunit (p110β) of PI3Kβ whose mutation disrupts binding to Rab5. To better define the Rab5-p110β interface, we performed alanine-scanning mutagenesis and analyzed Rab5 binding with an in vitro pulldown assay with GST-Rab5GTP Of the 35 p110β helical domain mutants assayed, 11 disrupted binding to Rab5 without affecting Rac1 binding, basal lipid kinase activity, or Gβγ-stimulated kinase activity. These mutants defined the Rab5-binding interface within p110β as consisting of two perpendicular α-helices in the helical domain that are adjacent to the initially identified Gln596 and Ile597 residues. Analysis of the Rab5-PI3Kβ interaction by hydrogen-deuterium exchange MS identified p110β peptides that overlap with these helices; no interactions were detected between Rab5 and other regions of p110β or p85α. Similarly, the binding of Rab5 to isolated p85α could not be detected, and mutations in the Ras-binding domain (RBD) of p110β had no effect on Rab5 binding. Whereas soluble Rab5 did not affect PI3Kβ activity in vitro, the interaction of these two proteins was critical for chemotaxis, invasion, and gelatin degradation by breast cancer cells. Our results define a single, discrete Rab5-binding site in the p110β helical domain, which may be useful for generating inhibitors to better define the physiological role of Rab5-PI3Kβ coupling in vivo.


Design, synthesis and anticancer activity of N-aryl indolylsulfoximines: Identification of potent and selective anticancer agents.
A facile and efficient approach utilizing copper-mediated cross-coupling reaction of N-boc-3-indolylsulfoximines with aryl iodides was developed to synthesize a diverse range of N-arylated indolylsulfoximines 11a-m in excellent yields (up to 91%). The key precursors, free NH sulfoximines 9 were readily prepared by the treatment of N-boc-3-methylthioindoles 8 with a combination of IBD and ammonium carbamate. Under similar conditions NH-free indolylsulfoximine 9a was successfully prepared in gram-scale quantities. The reaction is highly chemoselective and tolerant of a wide range of functional groups. The process is environmentally friendly and is amenable to scale-up. Among the prepared N-arylated indolylsulfoximines 11a-m, compounds 11i-j (2.68-2.76 μM), 11f-g (1.9-3.7 μM) and 11k (1.28 μM) showed potent and selective cytotoxicity against 22Rv1, C4-2 and MCF7 cells, respectively. Indolylsulfoximine derivative 11l displayed a broad spectrum of activity (1.7-8.2 μM) against the tested cancer cell lines. These compounds were found to be non-cytotoxic to normal HEK293 cells, indicating their potential selectivity for cancer cells. We analysed the impact of 11l on various cellular assays to uncover its mechanism of action. Cellular assay shows that 11l increases the endogenous level of ROS, leading to the increased level of p-53 and c-jun inducing apoptosis. 11l also induced mitochondrial dysfunction, further promoting apoptotic pathways. Besides, 11l also restricts cell invasiveness, indicating that it could serve as an effective anti-metastatic agent. As oxidative stress severe F actin causing tubulin depolymerization, we examined the impact of 11l on tubulin dynamics. Accordingly, 11l treatment decreased the levels of polymerized tubulin in 22Rv1 and C4-2 cells. Although future studies are needed to determine their exact molecular target(s), our data shows that N-aryl indolylsulfoximines could serve as effective anti-cancer agents.


Mentalizing partially mediates the association between attachment insecurity and global stress in preservice teachers.
Considering the fact that the teaching profession is a highly stressful occupation and that teachers' ineffective coping strategies contribute to higher levels of stress, the objective of the present study was to investigate whether insecure attachment is related to global stress experiences in preservice student teachers. Furthermore, it was examined whether this link is mediated by the preservice teachers' mentalizing-the capacity to perceive and consider one's own and others' behavior based on intentional mental states. Data of this cross-sectional study came from 202 preservice student teachers using self-report instruments (attachment: Expectation in Close Relationships; mentalizing: Reflective Functioning Questionnaire; stress: Trier Inventory of Chronic Stress). The hypotheses were tested using structural equation modelling. Anxious attachment was positively related to stress and impairments in mentalizing. In addition, the link between attachment-related anxiety and stress was partially mediated by mentalizing. Avoidant attachment was not associated with stress or mentalizing. Results indicate that anxious attachment is associated with higher perceived stress in preservice student teachers. Consequently, attachment-related anxiety may be a risk factor which, in turn, may foster perceptions of higher global stress experiences. Additional research needs to focus on exploring the link between attachment insecurity and global stress experiences among preservice student teachers. An examination of preservice student teachers' own attachment experiences proves to be useful, for example in the context of mentalization-based supervision or reflective practice.