TNF-alpha promotes cell survival through stimulation of K+ channel and NFkappaB activity in corneal epithelial cells.
Tumor necrosis factor (TNF-alpha) in various cell types induces either cell death or mitogenesis through different signaling pathways. In the present study, we determined in human corneal epithelial cells how TNF-alpha also promotes cell survival. Human corneal epithelial (HCE) cells were cultured in DMEM/F-12 medium containing 10% FBS. TNF-alpha stimulation induced activation of a voltage-gated K+ channel detected by measuring single channel activity using patch clamp techniques. The effect of TNF-alpha on downstream events included NFkappaB nuclear translocation and increases in DNA binding activities, but did not elicit ERK, JNK, or p38 limb signaling activation. TNF-alpha induced increases in p21 expression resulting in partial cell cycle attenuation in the G1 phase. Cell cycle progression was also mapped by flow cytometer analysis. Blockade of TNF-alpha-induced K+ channel activity effectively prevented NFkappaB nuclear translocation and binding to DNA, diminishing the cell-survival protective effect of TNF-alpha. In conclusion, TNF-alpha promotes survival of HCE cells through sequential stimulation of K+ channel and NFkappaB activities. This response to TNF-alpha is dependent on stimulating K+ channel activity because following suppression of K+ channel activity TNF-alpha failed to activate NFkappaB nuclear translocation and binding to nuclear DNA.


Primary structural modules of the dorsal nuclei of the thalamus and motor cortex in humans.
The aim of the present work was to study the primary structure of the modular organization of neurons in the dorsal nucleus of the thalamus (ventral anterior, ventral lateral, anterior ventral, anterior medial, anterior dorsal, medial, reticular) and motor cortex (fields 4 and 6) in humans by the Nissl, Kluver-Barrera, and Golgi silver nitrate impregnation methods. These studies showed that neurons in the dorsal nuclei of the thalamus and the motor cortex are organized as small groups and short chains fulfilling defined criteria. These neuronal complexes, forming individual structural units, were termed primary structural modules. Comparison of primary structural modules in the dorsal nuclei and motor cortex showed that the principles of organization were identical in both locations. Thus, the dorsal nuclei of the thalamus and motor cortex in humans show two principles of the organization of neurons into primary structural modules: the group and chain principles.


Prosthetic rehabilitation of edentulous patients with cleft palate.
In this work a description of prosthetic treatment of the edentulous patient with cleft palate was showed. Although positive results of surgical treatment diminish the number of these patients, more than once a dentist has to treat effectively patients in their fifties, sixties and seventies whose cleft palate was not treated with modern methods. Authors describe prosthetic solution which enables removing part of acryl from prosthesis and making air chambers. Diminishing of obturator prosthesis mass increases stabilization and retention and contributes to easier adaptation of prosthetic restoration.


Engineering Hybrid Chemotaxis Receptors in Bacteria.
Most bacteria use transmembrane sensors to detect a wide range of environmental stimuli. A large class of such sensors are the chemotaxis receptors used by motile bacteria to follow environmental chemical gradients. In Escherichia coli, chemotaxis receptors are known to mediate highly sensitive responses to ligands, making them potentially useful for biosensory applications. However, with only four ligand-binding chemotaxis receptors, the natural ligand spectrum of E. coli is limited. The design of novel chemoreceptors to extend the sensing capabilities of E. coli is therefore a critical aspect of chemotaxis-based biosensor development. One path for novel sensor design is to harvest the large natural diversity of chemosensory functions found in bacteria by creating hybrids that have the signaling domain from E. coli chemotaxis receptors and sensory domains from other species. In this work, we demonstrate that the E. coli receptor Tar can be successfully combined with most typical sensory domains found in chemotaxis receptors and in evolutionary-related two-component histidine kinases. We show that such functional hybrids can be generated using several different fusion points. Our work further illustrates how hybrid receptors could be used to quantitatively characterize ligand specificity of chemotaxis receptors and histidine kinases using standardized assays in E. coli.


Anti-Inflammatory Effects of Nutritionally Relevant Concentrations of Oleuropein and Hydroxytyrosol on Peripheral Blood Mononuclear Cells: An Age-Related Analysis.
Immunosenescence and inflammaging facilitate the insurgence of chronic diseases. The Mediterranean diet is a non-invasive intervention to improve the chronic low-grade inflammatory status associated with aging. Olive oil oleuropein (OLE) and hydroxytyrosol (HT) demonstrated a controversial modulatory action on inflammation in vitro when tested at concentrations exceeding those detectable in human plasma. We studied the potential anti-inflammatory effects of OLE and HT at nutritionally relevant concentrations on peripheral blood mononuclear cells (PBMCs) as regards cell viability, frequency of leukocyte subsets, and cytokine release, performing an age-focused analysis on two groups of subjects: Adult (age 18-64 years) and Senior (age ≥ 65 years). OLE and HT were used alone or as a pre-treatment before challenging PBMCs with lipopolysaccharide (LPS). Both polyphenols had no effect on cell viability irrespective of LPS, but 5 µM HT had an LPS-like effect on monocytes, reducing the intermediate subset in Adult subjects. OLE and HT had no effect on LPS-triggered release of TNF-α, IL-6 and IL-8, but 5 µM HT reduced IL-10 secretion by PBMCs from Adult vs. Senior group. In summary, nutritionally relevant concentrations of OLE and HT elicit no anti-inflammatory effect and influence the frequency of immune cell subsets with age-related different outcomes.


[Hygienic and toxicological safety assessment of nano- and micro-dispersed manganese oxide (III, IV)].
The findings of the experimental study performed within the preliminary hygienic and toxicological assessment of nanodispersed manganese oxide (III, IV), which was endogastrics administered to white nonlinear mice once, showed that, under acute oral exposure to manganese oxide, nanodispersed particles were 2,6-fold more toxic and cumulative (class 3, according to the oral LD50 values) than dispersed solution with micro-sized particles (0.4-592 microm, class 4). Endogactric exposure to nanodispersed solution of manganese oxide (III, IV) in organisme produced such negative health effects as vasodilation and plethora, pathological inclusions and increased cell aggregation, which increased and became more prevalent with higher doses. The prediction of chronic toxicity parameters demonstrated that threshold and NOAEL doses for nanodispersed solution were 1.6 and 0.26 mg/kg, respectively, which are 2.3 times lower than for finely divided solution of manganese oxide (III, IV).


Principles of health care reform: an African-American perspective.
To protect the interests of underserved Americans, fair and rational health care reform must embrace 10 principles. These include universal and comprehensive coverage; mandatory cost containment; equity; freedom to change jobs or to relocate; high quality; reduced paperwork; primary care; help for the underserved; consumer-oriented care; and fair financing.


Mechanochemical Synthesis and Characterization of Metastable Hexagonal Li4SnS4 Solid Electrolyte.
A new crystalline lithium-ion conducting material, Li4SnS4 with an ortho-composition, was prepared by a mechanochemical technique and subsequent heat treatment. Synchrotron X-ray powder diffraction was used to analyze the crystal structure, revealing a space group of P63/ mmc and cell parameters of a = 4.01254(4) Å and c = 6.39076(8) Å. Analysis of a heat-treated hexagonal Li4SnS4 sample revealed that both lithium and tin occupied either of two adjacent tetrahedral sites, resulting in fractional occupation of the tetrahedral site (Li, 0.375; Sn, 0.125). The heat-treated hexagonal Li4SnS4 had an ionic conductivity of 1.1 × 10-4 S cm-1 at room temperature and a conduction activation energy of 32 kJ mol-1. Moreover, the heat-treated Li4SnS4 exhibited a higher chemical stability in air than the Li3PS4 glass-ceramic.


Effects of REM deprivation and an NMDA agonist on the extinction of conditioned fear.
Rapid eye movement sleep (REM) has been implicated in a number of learning and memory tasks. Previous research has demonstrated that REM deprivation impairs the development of extinction of conditioned fear responses. However, the neurobiological mechanisms of this effect remain unclear. The present study investigated the effects of systemic administration of d-cycloserine (DCS), an NMDA agonist, on the extinction of a conditioned fear response following 6 h of REM deprivation. In experiment 1, rats were administered DCS between fear training and REM deprivation. In experiment 2, rats were administered DCS prior to extinction training. The results of experiment 1 indicated that both DCS alone and REM deprivation alone impaired extinction learning. Administration of DCS to REM deprived animals partially, but not completely, reversed the deficit in extinction. The results of experiment 2 indicated that regardless of prior REM deprivation history, DCS facilitated extinction learning. The results provide further evidence for a role of REM in the extinction of cued fear learning and indicate that this effect appears to be partially mediated by NMDA-dependent mechanisms.


Occurrence of diffuse, poorly differentiated hepatocellular carcinoma during pegylated interferon plus ribavirin combination therapy for chronic hepatitis C.
Interferon therapy is indicated for the treatment of chronic hepatitis C and prevention of hepatocellular carcinoma. We describe the case of a 66-year-old Italian woman who received pegylated interferon alpha-2a plus ribavirin combined therapy for HCV-related chronic liver disease. Preliminary hematochemical, ultrasound and bioptic investigations did not show liver cirrhosis or hepatocarcinoma. After 24 weeks of treatment transaminase serum levels were in the normal range and circulating HCVRNA was undetectable by PCR qualitative assay. On week 46 a serious adverse event occurred, with rapid transaminase increase, severe hyperpyrexia, and abdominal pain, leading to interruption of interferon and ribavirin. Liver biopsy was repeated and it revealed poorly differentiated hepatocellular carcinoma. Only palliative care could be performed and the patient died of liver failure within 2 months. The present case underlines that hepatocellular carcinoma can be misdiagnosed in spite of laboratory and instrumental follow-up. More sensitive tools are needed for tumor detection, to avoid IFN impairment of the liver, even though it eradicates HCV.


[Acute lower gastrointestinal tract bleeding caused by Dieulafoy lesion in rectum--case report].
Dieulafoy lesion may be one of the rare causes of acute gastrointestinal (GI) bleeding. This pathology mostly appears in the stomach wall, but very rarely it can be found in other parts of GI tract. Authors describe a case of 57-year-old female patient with a history of ulcerative colitis, staying in the complete remission since the moment of diagnosis many years ago, without any treatment. During the week before admission to the hospital, patient observed streaks of blood in the stool and felt some pain in the lower abdomen. On the day of admission to one of surgical departments in Szczecin, acute rectal bleeding had started in the patient, causing very serious state. The primary diagnosis was: rectal bleeding caused by a flare of ulcerative colitis. Sigmoidoscopy was performed in Gastroenterology Departament and it showed a vessel coming out of the rectal wall (like Dleulafoy lesion), with symptoms of active bleeding, while the mucosa was unchanged. The hemoclipping was performed and bleeding has been stopped. There was no recurrent bleeding.


Induced somatic and germinal reversion of the white-spotted-1 insertional mutant phenotype in Drosophila melanogaster.
The white-spotted-1 (Wsp1) mutant of Drosophila melanogaster is characterized by the presence of an 8.7 kb retrotransposon (B104) inserted in the regulatory region of the white locus. The frequency of reversion in both somatic tissue and the germline after exposure to three different alkylating agents has been analysed. To determine if germinal revertants were induced by precise excision of the insertional element we analysed several phenotypic revertants using PCR and Southern blot techniques. The results indicate that, under our experimental conditions, the mutagens used did not induce excision of B104 in the white gene. In addition, the revertant phenotypes obtained were due to the existence of second site modifiers acting on expression of white. Such modifiers map near the white locus and, at least in one case, may correspond to suppressor-of-white-spotted.


Decreased Expression of CHST-12, CHST-13, and UST in the Proximal Interphalangeal Joint Cartilage of School-Age Children with Kashin-Beck Disease: an Endemic Osteoarthritis in China Caused by Selenium Deficiency.
The objective of this study is to investigate changes in the expression of enzymes involved in chondroitin sulfate (CS) sulfation in distal articular surface of proximal interphalangeal joint isolated from school-age children patients with Kashin-Beck disease (KBD), using normal children as controls. Articular cartilage samples were collected from four normal and four KBD children (7-12 years old), and these children were assigned to control and KBD groups. Hematoxylin and eosin (H&E), toluidine blue (TB), and immunohistochemical (IHC) stainings were utilized to evaluate changes in joint pathology and expression of enzymes involved in CS sulfation, including carbohydrate sulfotransferase 12 (CHST-12), carbohydrate sulfotransferase 13 (CHST-13), and uronyl 2-O-sulfotransferase (UST). The correspondence results were examined by semi-quantitative analysis. Compared with the control group, the KBD group showed the following: a significant decrease of total chondrocytes in superficial, middle, and deep layers and deposition of sulfated glycosaminoglycans in extracellular matrix of KBD cartilage were observed; positive staining chondrocytes of CHST-12, CHST-13, and UST were significantly less in superficial zone of KBD cartilage; and CHST-13 positive staining chondrocytes was reduced in deep zone of KBD cartilage. In contrast, the positive staining rates of CHST-12, CHST-13, and UST in KBD were significantly higher than those in the control group. The decreased expression of these enzymes and the physiologic compensatory reaction may be the signs of early-stage KBD. The alterations of CS structure modifying sulfotransferases in finger articular cartilage might play an important role in the onset and pathogenesis of school-age KBD children.


A crowbarless power supply for klystrons.
A new crowbarless power supply is to be installed at the New SUBARU storage ring. A high-power switching inverter unit eliminates the need for expensive and unstable crowbar circuits for the klystron power supply. It also realizes a very small voltage ripple in the low-frequency region. This is an important characteristic, especially in a quasi-isochronous storage ring such as New SUBARU.


Impact of physical activity on patient self-reported outcomes of lifelong premature ejaculation patients: Results of a prospective, randomised, sham-controlled trial.
Previous studies have investigated whether physical activity increases serotonin hormone levels. Serotonin receptor dysfunction is one of the frequently accused factors of premature ejaculation (PE). Nevertheless, no studies up to date have demonstrated that the association between physical activity and premature ejaculation. We aimed to investigate the relationship between physical activity and PE and determine whether moderate physical activity might delay ejaculation time or be an alternative treatment for PE. A total of 105 patients diagnosed with PE were enrolled in this study. Of the patients, 35 were treated with dapoxetine, (30 mg) on demand (Group 1), 35 performed moderate physical activities (Group 2), and 35 performed minimal physical activity (Group 3-sham). Demographic characteristics, metabolic equivalents (MET), premature ejaculation diagnostic tool (PEDT) and intravaginal ejaculatory latency time (IELT) were recorded. There were no significant differences among three groups in terms of age, BMI, MET, PEDT or IELT before treatment. At the end of the study, there was significant decrease in PEDT scores, and increase in IELT in groups 1 and 2 as compared to Group 3. In conclusion, a moderate physical activity longer than 30 min at least 5 times a week leads to ejaculation delay, and appears as an alternative to dapoxetine on demand for the treatment of PE.


Mandibular reconstruction in 2004: an analysis of different techniques.
The field of mandibular reconstruction has evolved dramatically over the past fifty years. Numerous advances in microsurgical technique, plating technology and instrumentation, and an understanding of donor site angiosomes have made consistent and reliable mandibular reconstruction possible. Refinements in technique continue to improve the functional and aesthetic outcomes of oromandibular reconstruction. This review discusses the current state-of-the-art techniques for mandibular reconstruction and highlights the latest innovations in technique. The most common indication for oromandibular reconstruction remains ablative surgery for advanced neoplastic processes of the oral cavity and oropharynx. Reconstruction of these complex three-dimensional composite bony and soft-tissue defects is paramount for rehabilitation of form and function. Vascularized osseous free tissue transfer is the state-of-the-art for mandibular reconstruction. The long-term excellent functional and aesthetic outcomes of this technique have recently been reported. The most commonly used free flaps for mandibular reconstruction are the fibula, iliac crest, and scapula. Each of these typically accepts endosseous implants improving functional outcomes. The use of mandibular reconstruction plates and coverage with a soft-tissue flap remains a reconstructive option for selected patients. The latest refinements in technique include temporary intraoperative external fixation, the use of periosteal free flaps, distraction osteogenesis, and development of biodegradable biopolymer scaffolds for mandibular defects. Oromandibular reconstruction, although a challenge for the head and neck reconstructive surgeon, is now reliable and highly successful with excellent long-term functional and aesthetic outcomes.


Reviewing the evidence for using continuous subcutaneous metoclopramide and ondansetron to treat nausea & vomiting during pregnancy.
To examine the medical evidence regarding the clinical efficacy and cost-effectiveness of the application of continuous subcutaneous metoclopramide and ondansetron to treat nausea and vomiting during pregnancy. All of the published peer-reviewed articles on the subject were assembled and assigned a level of evidence based on research design. The search uncovered one level II matched, controlled trial and three level III uncontrolled, retrospective case series published in peer review journals, as well as a book chapter. The book chapter, although not subjected to the peer-review process, is included in this review due to the paucity of other evidence. The matched cohort trial showed that continuous subcutaneous metoclopramide is significantly less-tolerated than continuous subcutaneous ondansetron (31.8% vs. 4.4%; P < 0.001). The four case series reported complete symptom resolution for 63.9% to 75% of the patients. Complications arose in 24.9% to 30.5% of the selected cases that were severe enough to require discontinuation of therapy. Complications included side effects of a worsening of symptoms. All of the trials are retrospective and observational in nature and, therefore, subject to the limitations inherent in the research design. Absent the benefit of meaningful cohort controls, comparative statements effectiveness cannot be substantiated with the available data. Randomized, controlled trials of sufficient power are necessary before long-term continuous subcutaneous metoclopramide or ondansetron can be used on a widespread basis to treat nausea and vomiting during pregnancy. Cost approximations in the case series are reported and, when compared to the cost of other methods of treatment previously published in the medical literature, the therapy appears to be cost-prohibitive. However, definitive statements cannot be made regarding cost-effectiveness until clinical efficacy is demonstrated through a sufficiently powered, well-designed, randomized control trial (RCT). Until such time, the therapy should remain experimental and coverage be restricted to intractable hyperemesis gravidarum (HG) that is unresponsive to more-conventional treatment options.


Implication of complex vertebral malformation and bovine leukocyte adhesion deficiency DNA-based testing on disease frequency in the Holstein population.
Two inherited lethal disorders, bovine leukocyte adhesion deficiency (BLAD) and complex vertebral malformation (CVM), play a major role in breeding of Holstein cattle. Both inherited diseases are based on single nucleotide polymorphisms that have been known for 12 and 7 yr, respectively. A total of 25,753 cattle were genotyped for BLAD (18,200 tests) and CVM (14,493 tests) in our laboratory since the beginning of the genotyping programs for these diseases. Based on founder effects, the CVM mutation is thought to be linked to milk production. The BLAD was genotyped using RFLP until 2001; then a fluorescence resonance energy transfer assay on a LightCycler was used, as for CVM genotyping. By using single nucleotide polymorphism-aided breeding, the allelic frequency of the BLAD and CVM mutations in the active sire population was reduced from 9.4% in 1997 to 0.3% in 2007 (BLAD) and from 8.3% in 2002 to 2.3% in 2007 (CVM), with calculated half-life of the mutant allele of 2.1 yr for BLAD and 3.6 yr for CVM. An observed increase of BLAD frequency in 1999 could be attributed to the massive use of a BLAD-positive sire tested falsely negative in another laboratory. These data show that marker-assisted selection is capable of substantially reducing the frequency of a mutation within a period of not more than 5 yr. The different selection strategies against the lethal recessive allele in CVM and BLAD are reflected in the different reduction rates of the specific allele frequencies.


A proposed blind routine quality control system for multichannel analyzers.
A blind quality control system using a special type of commercial reference serum is described. As packaged, the product contains six pools of analyzed reference sera with vials identified by coded numbers. The test constituents in the pools cover the range of multichannel analyzers in a random manner, i.e., each pool contains constituents in the low, normal and elevated ranges. As specific pools are not identified, and the analyst is unaware of the expected values for each constituent, results collected for quality control purposes are unbiased, i.e., free of conscious or unconscious editing. Data analysis in the proposed system is by least-squares technics. During an eight-week period, performance was studied using the standard error of the estimate to evaluate precision and the slope of the regression line to evaluate accuracy on the SMA and Mark X instruments. As an empirical comparison, the data are expressed as relative errors and precision and accuracy of analyzer performance evaluated as mean and standard deviation. The proposed regression analysis-based technic has the advantage of simultaneously evaluating linearity, as well as checking accuracy and precision over the total dynamic range of the instrument by use of a series of related serum pools.


Preparation and Characterization of Group 13 Cyanides.
Binary Group 13 cyanides [PPh4 ][Ga(CN)4 ], [PPh4 ]2 [In(CN)5 ], and [PPh4 ]2 [Tl(CN)5 ] were obtained in quantitative yields from the corresponding metal trifluorides MF3 (M=Ga, In, Tl) by fluoride-cyanide exchange reactions with Me3 SiCN in the presence of stoichiometric amounts of [PPh4 ]CN in acetonitrile solution. The 2,2'-bipyridine (bipy) adducts [(bipy)2 Ga(CN)2 ][Ga(CN)4 ], [(bipy)In(CN)3 ], and [(bipy)Tl(CN)3 ] were obtained from the reaction of MF3 with Me3 SiCN and bipy in acetonitrile. While the reaction of the metal trifluorides with Me3 SiCN in acetonitrile resulted in recovery of the starting materials, the reaction of MF3 with Me3 SiCN in pyridine (py) solution resulted in the formation of the pyridine adducts [(py)2 Ga(CN)3 ], [(py)3 In(CN)3 ], and [(py)2 Tl(CN)3 ]. The cyano compounds were characterized by their vibrational spectra and, in most cases, by their X-ray crystal structures.


The guanylate-binding proteins: emerging insights into the biochemical properties and functions of this family of large interferon-induced guanosine triphosphatase.
Originally identified by their unusual ability to bind guanosine monophosphate (GMP) nucleotide agarose, the guanylate-binding proteins (GBPs) were used extensively to promote our understanding of interferon-induced gene transcription and as markers of interferon responsiveness. Structural and biochemical analyses of human GBP-1 subsequently demonstrated that the GBPs are a unique subfamily of guanosine triphosphatase (GTPases) that hydrolyze guanosine triphosphate (GTP) to both guanosine diphosphate (GDP) and GMP. As members of the larger dynamin superfamily of GTPases, GBPs exhibit such properties as nucleotide-dependent oligomerization and concentration-dependent GTPase activity. Recently, progress has been made in assigning functions to members of the GBP family. While many of these functions involve protection against intracellular pathogens, a growing number of them are not directly related to pathogen protection. It is currently unclear how the unusual properties of GBPs contribute to this growing list of functions. As future studies uncover the molecular mechanism(s) of action of the GBPs, we will gain a greater understanding of how individual GBPs can mediate what currently appears to be a divergent set of functions.


Owner perceptions of their cat's quality of life when treated with a modified University of Wisconsin-Madison protocol for lymphoma.
Objectives The objectives of this study were to assess owner perceptions of their cat's quality of life during treatment for lymphoma with a doxorubicin-containing multi-agent chemotherapy protocol, whether various health-related parameters correlated with quality of life scores, and to assess owner satisfaction with the protocol. Methods A postal questionnaire was sent to the owners of 33 treated cats. Owners retrospectively assessed their cat's quality of life using a Likert scale (1-10) before lymphoma was diagnosed, at diagnosis and during chemotherapy. Owners assigned scores to various health-related parameters previously reported to affect quality of life at the three time points, and correlations with quality of life scores were sought. Owners were asked to rate the importance of these health-related parameters. Satisfaction with the protocol was investigated. Results Twenty questionnaires were completed (61% response rate). The median quality of life score before diagnosis (10, range 5-10) was higher than at diagnosis (3, range 1-9) ( P <0.05). The median quality of life score during chemotherapy (7, range 3-9) was lower than before diagnosis ( P <0.05) and higher than at diagnosis, but this was not statistically significant. Quality of life scores did not correlate with individual health-related parameter scores consistently; however, quality of life scores did correlate with appetite scores during chemotherapy. Appetite, vomiting and diarrhoea were parameters perceived as important in affecting quality of life. Most owners (75%) were happy they had treated their cat. Conclusions and relevance The quality of life scores observed were comparable to a previous study using cyclophosphamide, vincristine and prednisolone, employing the same scoring system. Although quality of life scores during chemotherapy were not significantly improved at diagnosis, owner satisfaction with the protocol was high. The factors perceived by owners to determine quality of life in their pets may be different to those previously conjectured, but appetite during chemotherapy remains important.


Conformations of nucleotides bound to wild type and Y78F mutant yeast guanylate kinase: proton two-dimensional transferred NOESY measurements.
Wild type and Y78F mutant yeast guanylate kinase (GKy) were studied to investigate the effects of a site-directed mutation on bound substrate conformations. Previously published work showed that Y78 is involved in GMP binding and that the Y78F mutant has 30-fold weaker GMP binding and 2 orders of magnitude less activity, than the wild type. Adenosine conformations of adenosine 5'-triphosphate (ATP) and adenosine 5'-diphosphate (ADP) and guanosine conformations of guanosine 5'-monophosphate (GMP) bound to wild type and Y78F mutant yeast guanylate kinase in the complexes GKy x Mg(II)ATP, GKy x Mg(II)ADP, GKy x GMP, and GKy x Mg(II)ADP x [U-13C]GMP were determined by two-dimensional transferred nuclear Overhauser effect (TRNOESY) measurements combined with molecular dynamics simulations. For adenyl nucleotides in wild type complexes, all glycosidic torsion angles, chi, were 54 +/- 5 degrees. In Y78F mutant complexes, adenyl nucleotide glycosidic torsion angles were 55 +/- 5 degrees (GKy x MgATP) and 49 +/- 5 degrees (GKy x MgADP). Thus, the adenyl nucleotides bind similarly for both the wild type and Y78F mutant complexes. However, in the fully constrained, two-substrate complexes, GKy x Mg(II)ADP x [U-13C]GMP, the guanyl glycosidic torsion angle, chi, is 50 +/- 5 degrees with the wild type and 83 +/- 5 degrees with the Y78F mutant. This difference suggests that an unfavorable torsion may be a large part of the mechanism for significantly weaker GMP binding to reaction complexes of the Y78F mutant.


Metabolic syndrome in HIV-infected middle-aged women on antiretroviral therapy: prevalence and associated factors.
To determine the prevalence of metabolic syndrome (MetS) and its associated factors in a group of HIV-infected middle-aged women. Cross-sectional study including 273 HIV-infected climacteric women of 40-60 years of age under care in two HIV outpatient reference centers in Brazil. Metabolic syndrome diagnosis was based on 2006 International Diabetes Federation criteria. Sociodemographic, clinical and behavioral factors were evaluated as well as HIV infection-related parameters. Mean age was 47.7 years; 59.1% of women were premenopausal, 91% were on antiretroviral therapy. Current CD4 count was >500cells/mm3 in 61.7%, current viral load undetectable in 76.9% of women, and a quarter had previous diagnosis of aids. The prevalence of metabolic syndrome in the subgroup of menopausal women was 46.9%. Univariate analysis showed an association between metabolic syndrome and age ≥50 years (p=0.002), schooling <8 years (p=0.003), post-menopause (p<0.001), body mass index (BMI) >25kg/m2 (p<0.001), and FSH ≥40mIU/mL (p=0.002). In the multivariate analysis only increased BMI (PR=1.09; 95% CI: 1.05-1.13; p<0.001) and FSH levels ≥40mIU/mL (PR=1.66; 95% CI: 1.14-2.40; p=0.008) maintained statistical significance. There was no association between PI use or any other factor related to HIV-infection and MetS in any of the analyses performed. High BMI and FSH levels compatible with menopause were the only factors associated with MetS in these middle aged HIV-infected women. In the context of well-controlled, early treated HIV infection, traditional rather than HIV-related factors were associated with MetS.


VDAC regulates AAC-mediated apoptosis and cytochrome c release in yeast.
Mitochondrial outer membrane permeabilization is a key event in apoptosis processes leading to the release of lethal factors. We have previously shown that absence of the ADP/ATP carrier (AAC) proteins (yeast orthologues of mammalian ANT proteins) increased the resistance of yeast cells to acetic acid, preventing MOMP and the release of cytochrome c from mitochondria during acetic acid - induced apoptosis. On the other hand, deletion of POR1 (yeast voltage-dependent anion channel - VDAC) increased the sensitivity of yeast cells to acetic acid. In the present work, we aimed to further characterize the role of yeast VDAC in acetic acid - induced apoptosis and assess if it functionally interacts with AAC proteins. We found that the sensitivity to acetic acid resulting from POR1 deletion is completely abrogated by the absence of AAC proteins, and propose that Por1p acts as a negative regulator of acetic acid - induced cell death by a mechanism dependent of AAC proteins, by acting on AAC - dependent cytochrome c release. Moreover, we show that Por1p has a role in mitochondrial fusion that, contrary to its role in apoptosis, is not affected by the absence of AAC, and demonstrate that mitochondrial network fragmentation is not sufficient to induce release of cytochrome c or sensitivity to acetic acid - induced apoptosis. This work enhances our understanding on cytochrome c release during cell death, which may be relevant in pathological scenarios where MOMP is compromised.


Comparison of wear, wear debris and functional biological activity of moderately crosslinked and non-crosslinked polyethylenes in hip prostheses.
The wear, wear debris and functional biological activity of non-crosslinked and moderately crosslinked ultrahigh molecular weight polyethylene (UHMWPE) acetabular cups have been com pared when articulating against smooth and intentionally scratched femoral heads. Volumetric wear rates were determined in a hip joint simulator and the debris was isolated from the lubricant and characterized by the percentage number and volumetric concentration as a function of particle size. The volumetric concentration was integrated with the biological activity function determined from in vitro cell culture studies to predict an index of specific biological activity (SBA). The product of specific biological activity and volumetric wear rate was used to determine the index of functional biological activity (FBA). On smooth femoral heads the crosslinked UHMWPE had a 30 per cent lower wear rate, but it had a greater percentage volume of smaller, more biologically active particles, which resulted in a similar index of FBA compared with the non-crosslinked material. On the scratched femoral heads the volumetric wear rate was three times higher for the moderately crosslinked UHMWPE and two times higher for the non-crosslinked UHMWPE compared with the smooth femoral heads. This resulted in a higher wear rate for the moderately crosslinked material on the scratched femoral heads. All the differences in wear rate were statistically significant. There were only small differences in particle volume concentration distributions, and this resulted in similar indices of FBA which were approximately twice the values of those found on the smooth femoral heads. Both materials showed lower wear and FBA than for previously studied aged and oxidized UHMWPE gamma irradiated in air. However, this study did not reveal any advantage in terms of predicted FBA for moderately crosslinked UHMWPE compared with non-crosslinked UHMWPE.


MiR-185-3p mimic promotes the chemosensitivity of CRC cells via AQP5.
Studies showed that microRNAs (miRNAs) are important regulators in drug resistance. The current study investigated the role of miR-185-3p and its predicted target gene AQP5 in 5-FU-insensitive colorectal cancer (CRC) cells. Quantitative real-time polymerase chain reaction (qRT-PCR) and Spearman's correlation analysis were conducted to determine the correlation of expression levels of miR-185-3p and AQP5 from CRC tissues. HCT-116 and HCT-8 cells were treated by gradient concentration of 5-FU to construct 5-FU-resistant CRC model. The inhibition and viability of 5-FU-resistant cells were detected by MTT assay, and cell migration and invasion ability were determined by wound healing and transwell assay. The expressions of miR-185-3p and AQP5 were measured by qRT-PCR. StarBase and dual-luciferase reporter assay were used to predict and confirm the interaction between miR-185-3p and AQP5. Further experiments were performed to explore the function of miR-185-3p in 5-FU-resistant cells through regulating aquaporin-5 (AQP5). The levels of EMT-associated markers and AQP5 were determined by conducting Western Blot and qRT-PCR. We found that 5-FU-resistant CRC cells showed a lower inhibition rate, and higher migration and invasion abilities. MiR-185-3p was low-expressed in CRC tissues and 5-FU-resistance cells, and it targeted and regulated the expression of AQP5, which was found up-regulated in CRC and 5-FU-resistance CRC cells (r = -0.29, P < .05). Furthermore, miR-185-3p mimic enhanced the chemo-sensitivity of 5-FU-resistant cells, while overexpressed AQP5 reversed such an effect produced by miR-185-3p mimic. MiR-185-3p mimic enhances the chemosensitivity of CRC cells via AQP5. Our research provides a potential therapeutic target for 5-FU-resistant CRC cells.


The Accurate Free-hand Placement of S2 Alar Iliac (S2AI) Screw.
Achieving an osseous fusion across the lumbosacral spine is still challenging in spine surgery. For the long multisegmental fusion surgery, it is crucial to build a robust and substantial foundation, which sometimes necessitates the distal spinal fixation to the pelvis. The pelvic fixation technique involves advancing the screw through the alar, thereby providing more purchase across the sacroiliac joint and into the ilium. The S2 alar iliac screws can obtain immediate stability and proper biomechanical strength of constructs. Here we demonstrated our experience of how to place the S2 alar iliac screw accurately as well as some pitfalls and pearls of this technique.


Younger women's experiences of deciding against delayed breast reconstruction post-mastectomy following breast cancer: An interpretative phenomenological analysis.
Most women do not reconstruct their breast(s) post-mastectomy. The experiences of younger women who maintain this decision, although important to understand, are largely absent in the research literature. This interview-based study uses interpretative phenomenological analysis to explore the experiences of six women, diagnosed with primary breast cancer in their 30s/40s, who decided against delayed reconstruction. Findings reported here focus on one superordinate theme (decision-making) from a larger analysis, illustrating that the women's drive to survive clearly influenced their initial decision-making process. Their tenacity in maintaining their decision is highlighted, despite non-reconstruction sometimes being presented negatively by medical teams. Patient-centred support recommendations are made.


First report of the ectomycorrhizal fungal community associated with two herbaceous plants in Inner Mongolia, China.
Ectomycorrhizal (EM) fungi play a vital role in ensuring plant health, plant diversity, and ecosystem function. However, the study on fungal diversity and community assembly of EM fungi associated with herbaceous plants remains poorly understood. Thus, in our study, Carex pediformis and Polygonum divaricatum in the subalpine meadow of central Inner Mongolia, China were selected for exploring EM fungal diversity and community assembly mechanisms by using llumina MiSeq sequencing of the fungal internal transcribed spacer 2 region (ITS2). We evaluated the impact of soil, climatic, and spatial variables on EM fungal diversity and community turnover. Deterministic vs. stochastic processes for EM fungal community assembly were quantified using β-Nearest taxon index scores. The results showed that a total of 70 EM fungal OTUs belonging to 21 lineages were identified, of which Tomentella-Thelephora, Helotiales1, Tricholoma, Inocybe, Wilcoxina were the most dominant EM fungal lineages. EM fungal communities were significantly different between the two herbaceous plants and among the two sampling sites, and this difference was mainly influenced by soil organic matter (OM) content and mean annual precipitation (MAP). The neutral community model (NCM) explained 45.7% of the variations in EM fungi community assembly. A total of 99.27% of the β-Nearest Taxa Index (βNTI) value was between -2 and 2. These results suggest that the dominant role of stochastic processes in shaping EM fungal community assembly. In addition, RCbray values showed that ecological drift in stochastic processes dominantly determined community assembly of EM fungi. Overall, our study shed light on the EM fungal diversity and community assembly associated with herbaceous plants in the subalpine region of central Inner Mongolia for the first time, which provided a better understanding of the role of herbaceous EM fungi.


Boolean reconstructions of complex materials: Integral geometric approach.
We show that for the Boolean model of random composite media one can, from a single image of a system at any particle fraction, define a set of parameters which allows one to accurately reconstruct the medium for all other phase fractions. The morphological characterization is based on a family of measures known in integral geometry which provides powerful formulas for the Boolean model. The percolation thresholds of either phase are obtained with good accuracy. From the reconstructions one can subsequently predict property curves for the material across all phase fractions from the single three-dimensional image. We illustrate this for transport and mechanical properties of complex Boolean systems and for experimental sandstone samples.


The use of fetal fibronectin testing in the management of a triplet pregnancy with a short cervix.
Following in vitro fertilisation treatment, a 40-year-old woman was expecting trichorionic, triamniotic triplets. Her cervix shortened from 34 mm at 16(+5) weeks to 16 mm at 20(+5) weeks, a risk reported with 100% delivery before 28 weeks gestation. She was admitted to hospital and at 24(+1) weeks was given corticosteroids. From 21(+5) weeks her cervical length remained below 16 mm. However, weekly fetal fibronectin (fFN) tests were negative from 22(+5) weeks to delivery at 35(+5) weeks. This, along with an absence of symptoms, gave her doctors confidence to manage her as an outpatient from 28 weeks. At 33(+5) weeks she was diagnosed as having pre-eclampsia and three live births were delivered by prelabour caesarean section. Prior to delivery her cervical length was 10 mm and fFN test remained negative. There are no reports of outcome following a negative fFN with a short cervix in triplet pregnancies but fFN could be a useful tool, in conjunction with cervical length measurement, in the management of triplets.


[Spatial-epidemiological analysis of population aging in counties in Jiangsu Province, 1990-2020].
Objective: To analyze the space-time evolution trend of population aging at the county level in Jiangsu Province from 1990 to 2020. Methods: Taking each county of Jiangsu Province as the research object, we selected data from the fourth to the seventh national population censuses. We used the aging coefficient and age concentration rate as static and dynamic indicators to measure the degree of population aging, respectively. We also used spatial correlation analysis to study the spatial and temporal patterns and evolution characteristics of aging in counties in Jiangsu Province. Results: The degree of aging in each county of Jiangsu Province has been deepening, and the degree of aging gradually evolves from high in southern Jiangsu and low in northern Jiangsu to high in central Jiangsu and low in southern and northern Jiangsu and shows obvious spatial aggregation. From 1990 to 2020, the population aging in Jiangsu Province has accelerated, among which the aging from 2010-2020 is the fastest. The aging progress appeares the fastest in 2010-2020. Conclusion: From 1990 to 2020, population aging in Jiangsu Province has been deepening, with the fastest progress in the last decade and an obvious spatial aggregation.


[Acupuncture Reduces Acute Cerebral Ischemia by Suppressing Local and Peripheral Inflammation and Regulating Expression of Cerebral Protein Silent Information Regulator Factor 2-related Enzyme 1 and Nuclear Factor-κB in Rats].
To observe the effect of acupuncture on the expression of silent information regulator factor 2-related enzyme 1 (SIRT 1) and nuclear factor-κB (NF-κB) in rats with acute cerebral ischemia (ACI), so as to explore its mechanisms underlying improvement of ACI. One hundred female SD rats were randomly divided into 5 groups: normal control (normal), sham-operation (sham), model, non-acupoint and acupoint, with 20 rats in each. The ACI model was established by occlusion (electric coagulation) of the middle cerebral artery after craniotomy. "Baihui" (GV 20) and "Shuigou" (GV 26) or non-acupoints were punctured with filiform needles which were retained for 30 min after rotating for 1 min. The treatment was conducted once after modeling and 24 h thereafter. The cerebral infarct volume was measured after 2,3,5-triphenyltetrazolium chloride (TTC) staining. The contents of interleukin-1 β (IL-1 β), IL-6 and IL-8 in the serum and ischemic brain tissue were detected by radioimmunoassay, and the expression of protein SIRT 1 and NF-κB p 65 in the ischemic brain tissue was detected by immunoblotting. The brain infarction volume was obvious in the model group in comparison with the normal group (P<0.01), but was markedly reduced in the acupoint group relevant to the model group (P<0.05). The contents of IL-1 β, IL-6 and IL-8 in the serum and ischemic brain tissues, and the expression level of cerebral NF-κB p 65 protein were significantly increased (P<0.01), and the expression of cerebral SIRT 1 was obviously decreased in the model group compared with the normal group (P<0.01). Following acupuncture intervention, the increased IL-1 β, IL-6 and IL-8 contents and NF-κB p 65 expression were considerably down-regulated, and the decreased SIRT 1 expression was markedly up-regulated in the acupuncture acupoint group (P<0.05). No significant changes were found in the levels of all the indexes mentioned above between the normal and sham groups (P>0.05). Acupuncture intervention at acupoints can reduce the ischemic infarction volume in ACI rats, which may be associated with its effects in down-regulating the levels of IL-1 β, IL-6 and IL-8 in the serum and ischemic brain tissue, and in regulating cerebral SIRT 1/NF-κB signaling.


[Total endoprosthesis or dual head prosthesis in endoprosthetic management of femoral neck fractures?].
From 1.1.1984 to 31.12.1992, a total of 363 femoral neck fractures were treated by primary hip arthroplasty, with 213 total hip endoprostheses (58.7%) and 150 bipolar endoprostheses (41.3%). The average age of the 51 (14.0%) male and 312 (86.0%) female patients was 80.3 (+/- 8.9) years. While only 15.8% of all patients were treated with alloarthroplasty in 1984, the proportion treated in this way increased to 88.9% in 1992. In the same period, the percentage of patients with total hip arthroplasty who were above 80 years rose from 15.6% to 54.5% (p < 0.05), and the percentage in this group with multiple concomitant diseases rose from 28.1% to 48.2% (p < 0.05). General postoperative complications occurred after 43.5% of the operations (total arthroplasty 38.0%, bipolar prosthesis 51.3%, p < 0.01), with no substantial change during the observation period (1984-1986, 43.6%; 1990-1992, 43.2%). Surgical complications were observed in 9.1% of all cases (total arthroplasty 11.7%, bipolar prosthesis 5.6%, p < 0.05), decreasing slightly from 11.7% in 1984-1986 to 8.1% in 1990-1992. The postoperative mortality was 3.3% (total arthroplasts 1.9%, bipolar prosthesis 5.3%, p > 0.05). The higher rate of general complications and the insignificantly higher mortality after insertion of bipolar endoprostheses cannot be attributed to the endoprosthetic technique itself; they are probably due to the higher age and poorer general condition of patients in whom hemiarthroplasty was performed. Although there has been an increasing frequency of total hip replacement even in older and sicker patients in recent years, no significant changes in morbidity and mortality were observed.(ABSTRACT TRUNCATED AT 250 WORDS)


Racial/Ethnic Patterns in Opioid Dispensing among Medicaid-Funded Young Children.
Racial differences in opioid dispensing for diseases of the respiratory system (RESP) and injury (INJURY) outpatient visits among patients ≤ 3 years old were examined. Outpatient claims data of South Carolina Medicaid children were analyzed over three three-year periods. The variable of interest was the triennial rate of dispensed opioid prescriptions per 1000 visits for RESP and INJURY diagnoses across racial/ethnic groups. Overall, dispensed opioid prescription rates related to RESP declined for all racial/ethnic categories. White children had the highest dispensing rate for RESP indications in the first period (5.6), followed by Black (4.5), and Hispanic (4.1). The likelihood of White children being prescribed opioids was higher than Blacks, and this was persistent over the studied time (rate ratios from 1.24 to 1.22, respectively). Overall opioid dispensing rates related to injury declined during the studied time. Hispanics had the highest dispensing rate for INJURY (20.1 to 14.8 to 16.1, respectively) followed by White (16.1 to 13.1 to 10.4, respectively). Relative differences in the dispensing rates across groups increased over time (Hispanics vs. White: rate ratios from 1.25 to 1.55, Hispanics vs. Black: from 1.52 to 2.24, and White vs. Black: from 1.24 to 1.44, respectively). There are considerable differences in the dispensing rates across racial/ethnic groups, especially in injury-related prescribing.


Bloch surface waves assisted active modulation of graphene electro-absorption in a wide near-infrared region.
Light modulation has been recognized as one of the most fundamental operations in photonics. In this paper, we theoretically designed a Bloch surface wave assisted modulator for the active modulation of graphene electro-absorption. Simulations show that the strong localized electrical field generated by Bloch surface waves can significantly enhance the graphene electro-absorption up to 99.64%. Then by gate-tuning the graphene Fermi energy to transform graphene between a lossy and a lossless material, electrically switched absorption of graphene with maximum modulation depth of 97.91% can be achieved. Meanwhile, by further adjusting the incident angle to tune the resonant wavelength of Bloch surface waves, the center wavelength of the modulator can be actively controlled. This allows us to realize the active modulation of graphene electro-absorption within a wide near-infrared region, including the commercially important telecommunication wavelength of 1550 nm, indicating the excellent performance of the designed modulator via such mechanism. Such Bloch surface waves assisted wavelength-tunable graphene electro-absorption modulation strategy opens up a new avenue to design graphene-based selective multichannel modulators, which is unavailable in previous reported strategies that can be only realized by passively changing the structural parameters.


Sex Differences in the Use of Oral Anticoagulants for Atrial Fibrillation: A Report From the National Cardiovascular Data Registry (NCDR®) PINNACLE Registry.
Despite higher thromboembolism risk, women with atrial fibrillation have lower oral anticoagulation (OAC) use compared to men. The influence of the CHA2DS2-VASc score or the introduction of non-vitamin K OACs on this relationship is not known. Using the PINNACLE National Cardiovascular Data Registry from 2008 to 2014, we compared the association of sex with OAC use (warfarin or non-vitamin K OACs) overall and by CHA2DS2-VASc score and examined temporal trends in OAC use by sex. Multivariable regression models assessed the association between sex and OAC use in those with CHA2DS2-VASc scores ≥2. Temporal analyses assessed changes in OAC use by sex over time. Of the 691 906 atrial fibrillation patients, 48.5% were women. Women were significantly less likely than men to use any OAC overall (56.7% versus 61.3%; P<0.001) and at all levels of CHA2DS2-VASc score (adjusted risk ratio 9% to 33% lower, all P<0.001). Compared to other thromboembolic risk factors, female sex was associated with lower use of OAC (risk ratio 0.90, 95%CI 0.90-0.91). Over time, non-vitamin K OAC use increased at a slightly higher rate in women (56.2% increase per year, 95%CI 54.6% to 57.9%) compared to men (53.6% increase per year, 95%CI 52.0% to 55.2%), yet women remained less likely to receive any OAC at all time points (P<0.001). Among patients with atrial fibrillation, women were significantly less likely to receive OAC at all levels of the CHA2DS2-VASc score. Despite increasing non-vitamin K OAC use, women had persistently lower rates of OAC use compared to men over time.


Loin pain hematuria syndrome-visceral or neuropathic pain syndrome?
Loin pain hematuria syndrome (LPHS) is a rare pain syndrome, which is somewhat poorly characterized and challenging to treat. The condition of LPHS is still controversial and there is no consensus of validated diagnostic criteria or optimal treatment strategies. The epidemiology of LPHS is unknown and the mechanisms/etiologies contributing to LPHS remain uncertain. There exists some debate whether LPHS represents neuropathic pain or visceral pain. Curiously, for such a controversial pain syndrome there is substantially more written in the literature regarding surgical-type treatments than conservative treatments. A brief review of LPHS potential pathophysiology and potential treatment approaches is presented.


Energy expenditure and walking speed in lower limb amputees: a cross sectional study.
Energy expenditure and walking speed are generally recommended for use as measures of status and outcome for walking. The objective of this study was to measure the physiological cost index (PCI) and comfort walking speed (CWS) at three levels of lower limb amputation: transfemoral, transtibial and Syme level, and the relationship of these physiological variables to prosthetic ambulation supported with walking aids and stump length. This study was a prospective cross-sectional study.Eighty-nine individuals with lower limb amputation for reasons other than peripheral vascular disease (PVD) were recruited among patients at the Department of Prosthetics and Orthotics in University Clinical Center of Kosovo. PCI was assessed by five minutes of continuous indoor walking at CWS. Significant differences were found in PCI (F=29.87, P < 0.001) and CWS (F=19.33, P < 0.001) among the three amputation groups. Prosthetic ambulation supported with crutches showed an important impact on PCI (F=35.1, P < 0.001) and CWS (F=28.42, P < 0.001). Stump length was associated with significantly increased PCI (r=0.53, P = 0.02) and reduced CWS (r=0.58, P = 0.004) in transfemoral amputees. 1. A higher level of amputation is associated with less energy-efficient walking and with lower walking speed. 2. Prosthetic ambulation supported with crutches has significant impact on increasing of energy expenditure and decreasing walking speed. 3. Stump length is shown to have a major impact on PCI and CWS in transfemoral amputees.


Correlations between Serum P2X7, Vitamin A, 25-hydroxy Vitamin D, and Mycoplasma Pneumoniae Pneumonia.
Identifying new molecular diagnostic markers for Mycoplasma Pneumoniae Pneumonia (MPP) has always been an essential topic since MPP cases have increased every year, especially among children. Here, we examined the correlation between serum level of Purinergic receptor P2X7, vitamin A, and 25-hydroxy vitamin D (25(OH)D) and the severity of MPP, aiming to identify molecules that have the potential to become diagnostic markers. This study was conducted on 186 cases aged 1-14 (136 MPP and 50 non-MPP patients). Serum levels of Purinergic receptor P2X7, vitamin A, 25(OH)D, and multiple inflammatory and immune factors were measured, compared, and tested for statistical significance. Serum P2X7, tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) levels were significantly increased in severe MPP patients, while serum vitamin A, 25(OH)D, IgA, and IgG levels were significantly decreased. Our results demonstrated a positive correlation between serum P2X7 level and the severity of MPP, and negative correlations between serum levels of vitamin A and 25(OH)D and the severity of MPP, suggesting that high serum levels of P2X7 and low serum levels of vitamin A and 25(OH)D may indicate relatively severer MPP.


Structure and expression of the gene encoding murine M-protein, a sarcomere-specific member of the immunoglobulin superfamily.
The complete exon-intron organization of the murine gene encoding M-protein, a structural protein of sarcomeric myofibrils, was determined. The gene is composed of 37 exons and 36 introns, spanning approximately 75 kb of DNA. Intron positions are related to the modular structure of M-protein, which is composed essentially of immunoglobulin and fibronectin type III domains. Almost all repeats follow a two exon-one domain structure. The beginning and end of each domain are defined by introns in phase I; internal introns are more divergent in position and very rarely use phase I. A single transcriptional start point was detected in both skeletal and cardiac muscle. Analysis of the prospective promoter region revealed several potential regulatory elements. CAT expression assays using promoter deletion constructs identified three regions that seem to be most important for the muscle-specific transcription activation of the M-protein gene. These results provide the first complete characterization of a gene for a member of the intracellular branch of the immunoglobulin superfamily.


Host classification predicts infection after open fracture.
We propose that the classification commonly used in patients with osteomyelitis, the Gustilo classification, also is applicable to patients with open fractures as a method of identifying increased risk of infection because of comorbid medical illness. The records of 174 adult patients sustaining open fractures of long bones were retrospectively reviewed. Each patient was sorted into Class A, B, or C based on 14 immune system compromising factors. Class A has no compromising factors, Class B has one or two compromising factors, and Class C has more than three compromising factors. The association between class and infection was examined. The incidence of infection was 4%, 15%, and 30% in patients in Classes A, B, and C, respectively. Patients in Class B were 2.86 times more likely to have an infection, and patients in Class C were 5.72 times more likely to have an infection when both groups of patients were compared with patients in Class A. The Gustilo classification, location of fracture, and tobacco use are all factors associated with infection. Patients in Class B or C are at markedly increased risk of infection and may benefit from additional or alternative therapies that decrease the risk of infection. Prognostic study, Level II-1 (retrospective study). See the Guidelines for Authors for a complete description of levels of evidence.


Circulating permeability factor suPAR: from concept to discovery to clinic.
Kidney filtration functions through a hydrostatically driven multicellular barrier that includes visceral epithelial cells termed podocytes. These cells have delicate foot processes by which they ensure proper generation of urine. Most forms of chronic kidney disease start with disruption of podocytes, allowing for a breakdown of kidney barrier function and loss of plasma proteins from the blood into the urine (proteinuria). A classic podocyte disease is focal segmental glomerulosclerosis (FSGS) which progresses from podocyte injury to chronic kidney disease (CKD) and end-stage renal disease (ESRD) in a well-defined histopathological pattern. FSGS can recur after kidney transplantation in 30% or more cases, affecting both children and adults with rapid disease onset. This scenario has led credence to the concept that FSGS is caused by one or more circulating blood factor (s) that in turn cause podocyte injury. Decades of search for viable circulating permeability factor candidates have sparked research in this area. Recently, soluble urokinase plasminogen activator receptor (suPAR) was identified in a majority of patients with FSGS as a soluble factor that acts through binding to and activating podocyte beta 3 integrin. These observations show promise for novel therapeutic development to eliminate a portion of FSGS cases.


A protein-encapsulation technique by the sol-gel method for the preparation of monolithic columns for capillary electrochromatography.
A novel protein-encapsulation technique using sol-gels was developed for the preparation of monolithic capillary columns for capillary electrochromatography. Two chiral compounds, bovine serum albumin (BSA) and ovomucoid (OVM) from chicken egg white, were encapsulated in tetramethoxysilane-based hydrogel and their chiral selectivity was evaluated for the separation of some selected enantiomers (tryptophan, benzoin, eperisone, chlorpheniramine). The protein encapsulation was carried out within a capillary in a single step under mild conditions. The resultant monolithic columns showed adequate chromatographic performance, including mechanical strength, penetration of pressurized flow, and chiral separation. Two different proteins, BSA and OVM, were successfully encapsulated into the gel matrixes by changing the alkoxysilane compositions of the gel. Run-to-run repeatability was quite satisfactory. The consecutive analysis of the neutral compound, benzoin, by the OVM-encapsulated column showed good repeatability in the retention time (RSD = 1.23% for the first peak, N = 10). Under optimized conditions, the theoretical plate number for the first peak of benzoin reached 72,000 plates/m.


Increased IKKϵ protein stability ensures efficient type I interferon responses in conditions of TBK1 deficiency.
TBK1 and IKKϵ are related, crucial kinases in antiviral immune signaling pathways downstream of cytosolic nucleic acid receptors such as cGAS and RIG-I-like receptors. Upon activation, they phosphorylate the transcription factors IRF3 and IRF7 and thereby initiate the expression of type I interferons and antiviral effectors. While point mutation-induced loss of TBK1 kinase activity results in clinical hyper-susceptibility to viral infections, a complete lack of TBK1 expression in humans is unexpectedly not associated with diminished antiviral responses. Here, we provide a mechanistic explanation for these so-far unexplained observations by showing that TBK1 controls the protein expression of its related kinase IKKϵ in human myeloid cells. Mechanistically, TBK1 constitutively diminishes the protein stability of IKKϵ independent of TBK1 kinase activity but dependent on its interaction with the scaffold protein TANK. In consequence, depletion of TBK1 protein but not mutation-induced kinase deficiency induces the upregulation of IKKϵ. Due to the functional redundancy of the kinases in cGAS-STING and RIG-I-like receptor signaling in human myeloid cells, enhanced IKKϵ expression can compensate for the loss of TBK1. We show that IKKϵ upregulation is crucial to ensure unmitigated type I interferon production in conditions of TBK1 deficiency: While the type I interferon response to Listeria monocytogenes infection is maintained upon TBK1 loss, it is strongly diminished in cells harboring a kinase-deficient TBK1 variant, in which IKKϵ is not upregulated. Many pathogens induce TBK1 degradation, suggesting that loss of TBK1-mediated destabilization of IKKϵ is a critical backup mechanism to prevent diminished interferon responses upon TBK1 depletion.


Genetic variants in Chinese patients with sporadic Stanford type A aortic dissection.
Genetic disorders are strongly associated with aortic disease. However, the identities of genetic mutations in sporadic Stanford type A aortic dissection (STAAD) are not clear. The present study analysed the possible genetic mutations of the known pathogenic genes of aortic disease and the clinical characteristics in patients with sporadic STAAD. We analysed genetic mutations in 26 genes that underlie aortic aneurysms and dissections in 100 sporadic STAAD patients and 568 healthy controls after whole-genome sequencing (WGS). Clinical features and in-hospital death were determined in all STAAD patients. In total, 60 suspicious pathogenic mutations (56 novel and 4 previously reported) in 19 genes were identified in 50% (50/100) of patients, and 14 patients had more than 1 mutation. The ascending aortic diameter was extended in patients with mutations (49.1±12.3 vs. 43.7±11.2 mm, P=0.023), and the DeBakey type I phenotype was more common in patients with mutations in genes that coded extracellular matrix (ECM) components than in patients with mutations in other genes (96.6% vs. 66.7%, P=0.007). Patients with fibrillin-1 (FBN1) mutations were younger than patients without FBN1 mutations (44.7±11.0 vs. 53.5±12.1, P=0.030). Subgroup analyses revealed an increased risk of in-hospital mortality in mutation carriers (44.4% vs. 10.5%, P=0.029) but only in patients who received conservative treatment. Half of Chinese patients with a sporadic form of STAAD may carry mutations in known pathogenic genes of aortic disease, and these patients may exhibit distinct clinical features and poor clinical outcomes with the use of conservative treatment.


Rare complication of fibroids in pregnancy: Spontaneous fibroid rupture.
Uterine fibroids in pregnancy present with numerous complications. However, spontaneous rupture is a rare and potentially life-threatening event. We report a case of a 43-year-old multiparous woman who presented with hypovolemic shock secondary to a ruptured uterine fibroid 2 days after a spontaneous vaginal delivery. Emergency laparotomy confirmed massive intra-abdominal hemorrhage from the ruptured capsule of a 10-cm subserosal fibroid. A myomectomy was performed. Her postoperative recovery period was uneventful. This case is very rare, with fewer than 30 cases of hemoperitoneum secondary to rupture of fibroids having been reported in published works. It demonstrates how a benign and common condition, such as fibroids, can lead to an obstetric emergency that mimics a surgical abdomen requiring swift diagnosis and surgical intervention.


Medicinally Important Herbal Flowers in Sri Lanka.
The plant kingdom is rich with a numerous number of plants with various medical properties which can be used to treat various medical issues. Sri Lanka is a country full of biodiversity which is gifted with many plant resources. It has a rich history of traditional medicine systems consisting of Ayurveda, Unani, and Deshiya Chikitsa, where these plant resources are used as remedies for the diseases. In the traditional medicine system, various plant parts such as leaves, roots, fruits, flowers, and bark are used to treat disease conditions. Although less attention is paid to the medicinal importance of the flowers, some of them have been used to treat many diseases from the ancient time. Some properties of the flowers may differ from the properties of the other plant parts. For example, Sesbania grandiflora (Katuru murunga) flowers have shown anticancer properties against various cell models whereas some flowers have shown antispermatogenic properties. Flowers of Woodfordia floribunda (Militta) are added as fermenting agents in the preparation of Arishtas in Ayurveda. Also the most popular Clove oil is obtained from the flower buds of Syzygium aromaticum (Karabu-neti) which is used to treat toothaches since it has antibiotic and antiseptic properties. This article gives an overview of herbal flowers used in the traditional medicine system of Sri Lanka and their pharmacological importance. A comprehensive literature survey was done on the medicinally important flowers in Sri Lanka. Data was collected from Libraries of Ayurveda in Sri Lanka and from scientific databases. According to the survey many flowers are used as astringent, cardiac tonic, and febrifuge. Also some flowers are used to treat dysentery, diarrhoea, and indigestion. Some flowers are useful in the treatment of bleeding piles while some are useful in the treatment of asthma and bronchitis. It was revealed that there are many flowers with valuable therapeutic effects. Traditional medicine systems prevailing in Sri Lanka have made use of these flowers with therapeutic effects to cure so many diseases. The review of medicinally important herbal flowers provides knowledge and pharmacological leads which will help for the wellbeing of the human beings. Although there are phytochemical studies done to identify the chemical compounds on some flowers, chemical composition of many flowers remains unrevealed. So further studies need to be done to identify the chemical composition of these flowers.


[A 55-year-old man with prostate cancer, papilledema, and multiple cranial nerve palsies].
We report a 55-year-old man with papilledema and multiple cranial nerve palsies. He was well until 52 years of age when there was an onset of progressive difficulty in initiating urination; he visited the urology service of our hospital where a diagnosis of prostate cancer was made; the cancer was invading the bladder and was metastasizing to lymph nodes and bones. He was treated with oochiectomy and estrogen preparations with some improvement in his symptoms. Two years later, he developed difficulty in urination again, and transurethral resection of the tumor was performed in 1991. In December 1991, he noted tingling and numb sensation in his left face, which had become progressive worse within the next one month, and he developed blepharoptosis and deafness all on the left side. He was admitted to the urology service on February 4, 1992, and a neurological consultation was asked. On physical examination, general findings were unremarkable, except for lymph node enlargements of about 0.5 to 1.0 mm in size in cervical and inguinal regions. On neurologic examination, he was alert with normal mental activities; higher cerebral functions were intact. He had normal vision and visual fields, however, papilledema was present bilaterally; pupils and light reactions were normal. Extraocular muscles were intact on the right side, however, moderate restriction was noted in the left eye in that all the extraocular muscles except for the medial rectus were weak; blepharoptosis was noted on the left; no nystagmus was present. The sensation was diminished in the left face, and left facial paresis of the peripheral type was also noted; the taste sensation was also diminished in the left anterior two thirds of the tongue. He had sensorineural deafness on the left side. The other cranial nerves appeared intact. He walked normally; no weakness or muscle atrophy was noted; muscle tone was normal and no ataxia was observed. Deep reflexes were normally elicited and symmetric; the plantar response was flexor. No meningeal signs were present. Laboratory examination revealed following abnormalities: Hb 7.1 g/dl, platelet 47,000/cmm, WBC3,800/cmm, LDH 950IU/l, PAP232ng/ml (normal less than 1.6), PA2.631ng/ml (normal less than 7.4); a small amount of effusion was noted in the left pleural cavity; cytological examination of the fluid was class V. A cranial CT scan as well as MRI were entirely normal, as was the spinal tap. He was treated with glycerol, however, there was progressive increase in the pleural effusion, and he developed dyspnea; moist rale had become audible in the end of February.(ABSTRACT TRUNCATED AT 400 WORDS)


Regulation of cAMP concentration by calmodulin-dependent cyclic nucleotide phosphodiesterase.
Bovine brain contains two major calmodulin (CaM) dependent phosphodiesterase isozymes which are homodimeric proteins with subunit molecular masses of 60 and 63 kilodaltons (kDa), respectively. The 60-kDa subunit isozyme can be phosphorylated by cAMP-dependent protein kinase, resulting in a decrease in the enzyme affinity towards CaM. The phosphorylation is blocked by Ca2+ and CaM and reversed by the CaM-stimulated phosphatase (calcineurin). The 63-kDa subunit isozymes can also be phosphorylated, but in this case by a CaM-dependent protein kinase(s). This phosphorylation is also accompanied by a decrease in the isozyme affinity towards CaM and can be reversed by the CaM-dependent phosphatase. Analysis of the complex regulatory properties of the phosphodiesterase isozymes has led to the suggestion that fluxes of cAMP and Ca2+ during cell activations are closely coupled and that the CaM-dependent phosphodiesterase isozymes play key roles in this signal coupling phenomenon.


Metabolic targeting of lactate efflux by malignant glioma inhibits invasiveness and induces necrosis: an in vivo study.
Glioblastoma multiforme (GBM) are the most malignant among brain tumors. They are frequently refractory to chemotherapy and radiotherapy with mean patient survival of approximately 6 months, despite surgical intervention. The highly glycolytic nature of glioblastomas describes their propensity to metabolize glucose to lactic acid at an elevated rate. To survive, GBMs efflux lactic acid to the tumor microenvironment through transmembrane transporters denoted monocarboxylate transporters (MCTs). We hypothesized that inhibition of MCT function would impair the glycolytic metabolism and affect both glioma invasiveness and survival. We examined the effect on invasiveness with α-cyano-4-hydroxy-cinnamic acid (ACCA, 4CIN, CHCA), a small-molecule inhibitor of lactate transport, through Matrigel-based and organotypic (brain) slice culture invasive assays using U87-MG and U251-MG glioma cells. We then conducted studies in immunodeficient rats by stereotaxic intracranial implantation of the glioma cells followed by programmed orthotopic application of ACCA through osmotic pumps. Effect on the implanted tumor was monitored by small-animal magnetic resonance imaging. Our assays indicated that glioma invasion was markedly impaired when lactate efflux was inhibited. Convection-enhanced delivery of inhibitor to the tumor bed caused tumor necrosis, with 50% of the animals surviving beyond the experimental end points (3 months after inhibitor exhaustion). Most importantly, control animals did not display any adverse neurologic effects during orthotopic administration of ACCA to brain through programmed delivery. These results indicate the clinical potential of targeting lactate efflux in glioma through delivery of small-molecule inhibitors of MCTs either to the tumor bed or to the postsurgical resection cavity.


A review of quantitative studies of adherence to mental health clinical practice guidelines.
Mental health clinical practice guidelines have proliferated, but there is little evidence regarding the degree to which they are actually implemented in clinical practice. The goal of this study was to locate and review all peer-reviewed reports published through 2000 that provide quantitative information on rates of adherence to specific mental health guidelines. A literature search yielded 41 pertinent studies. These studies were of three types: 26 were cross-sectional investigations performed after the release of guidelines, six were conducted before and after release of guidelines without any specific intervention (pre/post), and nine involved a controlled trial of a specific intervention. Only 37% were conducted in the mental health specialty sector. Adequate adherence was found in 27% of the cross-sectional and pre/post studies but in 67% of the controlled trials. Successful interventions tended to be complex, involving system redesign or additional resources. Only six of 13 investigations (46%) that also measured patient outcome found that better outcome was associated with greater guideline adherence. Several studies showed that after cessation of interventions, adherence rates returned to preintervention levels. Thus, evidence indicates that guideline adherence is not high without specific intervention, but that certain interventions (typically multifaceted and resource-intensive ones) improve adherence. However, the public health challenge is to design and implement interventions that are sustainable in general clinical practice.


Optical mapping of pontine chemosensitive regions of neonatal rat.
We analyzed the neuronal response to hypercapnic acidosis, using an optical recording technique with a fluorescent voltage-sensitive dye (di-4-ANEPPS), in pontine slice preparations of neonatal rats, containing the locus coeruleus (LC), which has been electrophysiologically demonstrated to be chemosensitive. The dye-stained preparation was continuously superfused with artificial cerebrospinal fluid. Epifluorescence of the slice was detected using a high-sensitivity optical recording system. Changes in the intensity of fluorescence were serially analyzed while switching artificial cerebrospinal fluid from control to hypercapnic acidosis, or vice versa. The optical recording method revealed that the LC, as reported in previous studies, reversibly showed a depolarizing response to hypercapnic acidosis in 56% of the examined preparations. The A5 area (56%) also exhibited a reversible, depolarizing response to hypercapnic acidosis. The response was preserved under conditions in which chemical synaptic transmission was blocked by low Ca(2+)-high Mg(2+) solution. These results suggest that the optical recording method is applicable to identification of potentially chemosensitive areas, which deserve further electrophysiological analysis, and that the A5 area could be chemosensitive.


Targeting microspheres and cells to polyethylene glycol-modified biological surfaces.
It has previously been demonstrated that damaged arterial tissue can be acutely modified with protein-reactive polyethylene glycol (PEG) to block undesirable platelet deposition. This concept might be expanded by employing PEG-biotin and its strong interaction with avidin for site-specific targeted delivery. Toward this end, cultured endothelial cells (ECs) were surface modified with PEG-biotin and the available biotin was quantified with flow cytometry. NeutrAvidin-coated microspheres and PEG-biotin modified ECs with NeutrAvidin as a bridging molecule were delivered under arterial shear stress to PEG-biotin modified ECs on a coverslip as well as scrape-damaged bovine carotid arteries. After incubation with a 10 mM solution for 1 min, 8 x 10(7) PEG-biotin molecules/EC were found and persisted for up to 120 h. Perfused microspheres adhered to NHS-PEG-biotin treated bovine carotid arteries with 60 +/- 16 microspheres/mm(2) versus 11 +/- 4 microspheres/mm(2) for control arteries (p < 0.015). Similarly, 22 +/- 5 targeted ECs/mm(2) adhered to NHS-PEG-biotin treated bovine carotid arteries versus 6 +/- 2 ECs/mm(2) for control arteries (p < 0.01). The targeting strategy demonstrated here might ultimately find application for drug delivery, gene therapy, or cell therapy where localization to specific labeled vascular regions is desired following catheter-based or surgical procedures.


Laws Governing Nitrogen Loss and Its Numerical Simulation in the Sloping Farmland of the Miyun Reservoir.
Surface flow (SF) and subsurface flow (SSF) are important hydrological processes occurring on slopes, and are driven by two main factors: rainfall intensity and slope gradient. To explore nitrogen (N) migration and loss from sloping farmland in the Miyun Reservoir, the characteristics of total nitrogen (TN) migration and loss via SF and SSF under different rainfall intensities (30, 40, 50, 60, 70, and 80 mm/h) and slope gradients (5°, 10°, and 15°) were studied using indoor stimulated rainfall tests and mathematical models. Nitrogen loss via SF and SSF was found to increase exponentially and linearly with time, respectively, with SSF showing 14-78 times higher loss than SF. Under different rainfall intensities, SSF generally had larger TN loss loading than SF, thereby indicating that SSF was the main route for TN loss. However, the TN loss loading proportion via SF increasing from 14.03% to 35.82% with increasing rainfall intensity is noteworthy. Furthermore, compared with the measurement data, the precision evaluation index Nash-Suttcliffe efficient (NSE) and the determination coefficient (R2) of the effective mixing depth model in the numerical simulation of TN loss through SF in the sloping farmland in the Miyun Reservoir were 0.74 and 0.831, respectively, whereas those of the convection-dispersion equation for SSF were 0.81 and 0.811, respectively, thus indicating good simulation results. Therefore, this paper provides a reference for studying the mechanism of N migration and loss in sloping farmland in the Miyun Reservoir.


Squamous cell carcinoma of the prostate with strong FDG uptake on PET/CT.
Primary squamous cell carcinoma (SCC) of the prostate is a rare aggressive neoplasm with a poor prognosis. A 60-year-old man presented with urinary frequency and interrupted urination for 4 months. Serum prostate-specific antigen and SCC antigen were elevated. T2-weighted MR images showed a hyperintense lesion in the central area of the prostate. FDG PET/CT showed strong FDG uptake of the lesion with SUVmax of 15.5. Prostate biopsy revealed moderate differentiated SCC. This case indicates SCC of the prostate should be considered in the differential diagnosis of abnormal prostate FDG accumulation (especially with elevated SCC antigen level).


Crystal structure of AqpZ tetramer reveals two distinct Arg-189 conformations associated with water permeation through the narrowest constriction of the water-conducting channel.
AqpZ is a homotetramer of four water-conducting channels that facilitate rapid water movements across the plasma membrane of Escherichia coli. Here we report a 3.2 angstroms crystal structure of the tetrameric AqpZ (tAqpZ). All channel-lining residues in the four monomeric channels are found orientated in nearly identical positions with one marked exception at the narrowest channel constriction, where the side chain of a highly conserved Arg-189 adopts two distinct conformational orientations. In one of the four monomers, the guanidino group of Arg-189 points toward the periplasmic vestibule, opening up the constriction to accommodate the binding of a water molecule through a tridentate H-bond. In the other three monomers, the Arg-189 guanidino group bends over to form an H-bond with carbonyl oxygen of the Thr-183, thus occluding the channel. Therefore, the tAqpZ structure reveals two distinct Arg-189 confirmations associated with water permeation through the channel constrictions. Alternation between the two Arg-189 conformations disrupts continuous flow of water, thus regulating the open probability of the water pore. Further, the difference in Arg-189 displacements is correlated with a strong electron density found between the first transmembrane helices of two open channels, suggesting that the observed Arg-189 conformations are stabilized by asymmetrical subunit interactions in tAqpZ.


Changes in gain of vestibulospinal reflexes after local injection of beta-adrenergic substances in the cerebellar vermis of decerebrate cats.
Experiments were performed in decerebrate cats to find out whether the noradrenergic afferent system to the cerebellar cortex modified the gain of the vestibulospinal (VS) reflexes and, if so, whether this effect was mediated through beta-adrenoceptors. Unilateral injection into the vermal cortex of the culmen of the beta-adrenergic agonist isoproterenol (0.25-0.50 microliter at 8-16 micrograms/microliters of saline) increased the gain of the ipsilateral and to a lesser extent also of the contralateral triceps brachii to roll tilt of the animal at 0.15 Hz, +/- 10 degrees. The opposite result was obtained after local injection of an equal dose of the beta-adrenergic antagonist, propranolol. Only slight changes in the phase angle of the responses were obtained. The effective sites were located within the parasagittal zone B which upon stimulation inhibited the activity of the ipsilateral triceps brachii. Since most of the Purkinje (P)-cells of this corticocerebellar area, which project to the lateral vestibular nucleus, fire out-of-phase with respect to the related VS neurons, we postulate that the beta-adrenergic system enhances the amplitude of the P-cell responses to labyrinth stimulation, thus increasing the gain of the VS reflex.


Atherosclerotic stenosis of the renal arteries. Indications for intervention.
Advances in the ability to diagnose atherosclerotic stenosis of the renal arteries have increased awareness of this disorder over the past decade, and several medical specialties have become interested in its pathophysiology, diagnosis, and therapy. Improved surgical and anesthetic techniques and the development of endovascular technology have paved the way for more aggressive management of atherosclerotic renal artery stenosis. However, there is a lack of prospective data supporting this aggressive approach, and there is little agreement among physician groups concerning the appropriateness of revascularization in many patient scenarios. We will examine the available data and make recommendations about which patient groups might benefit most from renal artery revascularization (surgical or endovascular).


[Role of HIF signaling pathway involved in effects of hypoxia on proliferation and differentiation of myoblasts].
Oxygen is an environmental and developmental signal regulator, involved in energy homeostasis, development and process of differentiation. Myoblasts persist in skeletal muscle as satellite cells, which possess capability of self-renewing and differentiation into mature myofiber. Myoblasts play an important role in postnatal muscle regeneration after injury as well as maintaining myofibers function. Though oxygen is vital to nearly all forms of life, little is known on the effects of oxygen level on proliferation and differentiation of myoblasts. The level of oxygen in physiological and pathological microenvironment in vivo is low. This review summarizes recent advances in the study of the effects of hypoxia on proliferation and differentiation of myoblasts and HIF signaling pathway involved in these processes.


RoPE Score as a Predictor of Recurrent Ischemic Events After Percutaneous Patent Foramen Ovale Closure.
The benefits of patent foramen ovale (PFO) closure for cryptogenic stroke secondary prevention are still debated. The Risk of Paradoxical Embolism (RoPE) study developed a score to improve patient selection for this procedure. We proposed to assess the validity of this score to assess the prognostic impact of PFO closure.From 2000 to 2014, all consecutive patients submitted to PFO closure were included in a prospective registry in a university center. The primary endpoint was recurrent ischemic cerebrovascular events and the secondary endpoints were all-cause, neurological, and cardiac mortality rates and new-onset atrial fibrillation (NOAF) rates. In total, 403 patients were included in the study (women: 52.1%; mean age: 44.7 ± 10.9 years). The mean follow-up period was 6.4 ± 3.7 years. Immediate success was achieved in 97% patients. There were 23 (5.8%) ischemic cerebrovascular events, 8 (2.0%) deaths, and 17 (4.3%) NOAFs. The mean RoPE score was 6.10 ± 1.79. Smoker status, coronary artery disease, lower RoPE score, and higher left atrial dimensions were predictors of the primary endpoint. However, a lower RoPE score and coronary artery disease remained independent predictors in multivariate analysis.RoPE score was shown to be an independent predictor of recurrent ischemic cerebrovascular events, and a score of ≤ 6 was shown to identify patients with significantly higher risk of mortality and recurrent ischemic events.


The Hypnotic Induction in the Broad Scheme of Hypnosis: A Sociocognitive Perspective.
Researchers and clinicians typically divide hypnosis into two distinct parts: the induction and the suggestions that follow. We suggest that this distinction is arbitrary and artificial. Different definitions of hypnosis ascribe different roles to the hypnotic induction, yet none clearly specifies the mechanisms that mediate or moderate subjective and behavioral responses to hypnotic suggestions. Researchers have identified few if any differences in responding across diverse hypnotic inductions, and surprisingly little research has focused on the specific ingredients that optimize responsiveness. From a sociocognitive perspective, we consider the role of inductions in the broader scheme of hypnosis and suggest that there is no clear line of demarcation between prehypnotic information, the induction, suggestions, and other constituents of the hypnotic context. We describe research efforts to maximize responses to hypnotic suggestions, which encompass the induction and other aspects of the broader hypnotic framework, and conclude with a call for more research on inductions and suggestions to better understand their role within hypnotic interventions in research and clinical contexts.


Retinoid X receptor agonists impair arterial mononuclear cell recruitment through peroxisome proliferator-activated receptor-γ activation.
Mononuclear cell migration into the vascular subendothelium constitutes an early event of the atherogenic process. Because the effect of retinoid X receptor (RXR)α on arterial mononuclear leukocyte recruitment is poorly understood, this study investigated whether RXR agonists can affect this response and the underlying mechanisms involved. Decreased RXRα expression was detected after 4 h stimulation of human umbilical arterial endothelial cells with TNF-α. Interestingly, under physiological flow conditions, TNF-α-induced endothelial adhesion of human mononuclear cells was concentration-dependently inhibited by preincubation of the human umbilical arterial endothelial cells with RXR agonists such as bexarotene or 9-cis-retinoid acid. RXR agonists also prevented TNF-α-induced VCAM-1 and ICAM-1 expression, as well as endothelial growth-related oncogene-α and MCP-1 release. Suppression of RXRα expression with a small interfering RNA abrogated these responses. Furthermore, inhibition of MAPKs and NF-κB pathways were involved in these events. RXR agonist-induced antileukocyte adhesive effects seemed to be mediated via RXRα/peroxisome proliferator-activated receptor (PPAR)γ interaction, since endothelial PPARγ silencing abolished their inhibitory responses. Furthermore, RXR agonists increased RXR/PPARγ interaction, and combinations of suboptimal concentrations of both nuclear receptor ligands inhibited TNF-α-induced mononuclear leukocyte arrest by 60-65%. In vivo, bexarotene dose-dependently inhibited TNF-α-induced leukocyte adhesion to the murine cremasteric arterioles and decreased VCAM-1 and ICAM-1 expression. Therefore, these results reveal that RXR agonists can inhibit the initial inflammatory response that precedes the atherogenic process by targeting different steps of the mononuclear recruitment cascade. Thus, RXR agonists may constitute a new therapeutic tool in the control of the inflammatory process associated with cardiovascular disease.


Diet induced weight loss accelerates onset of negative alliesthesia in obese women.
The physiological and behavioral responses to hypocaloric diet are to increase energy intake to defend a steady body weight. We utilized the method of "negative alliesthesia" for measuring the hedonic reponse to sweet stimulus before (Initial session) and 3 months after entering a weight loss program. The negative alliesthesia test is known by physiologists but few clinical data exist. It is based on the observation that repeated pleasant gustatory stimuli turn into unpleasantness in the process of alliesthesia. At first visit participants repeatedly ingested sweet stimuli until they found them unpleasant and rated quantitatively on a linear analogue scale their hedonic experience. This procedure was repeated every 3 min until participants felt displeasure to end the session. The same protocol was followed after three months of following a weight loss diet. Dieting energy intake was from 1400 - 2000 kcal/d for 8 wk. Energy composition was 50% carb:25% prot: 25% lipid. After 8 wk caloric intake increased by 50 kcal/wk, to reach daily intake of 1800 - 2400 kcal/d. Energy composition was 50% carb:22% prot: 27% lipid. We report results on the effect of slow weight loss on negative alliesthesia in ten obese female participants enrolled in a commercial diet program based on Canada's Food Guide (Mincavi). Results showed that diet lowered the mean BMI (Initial session 36.8 +/- 1.8 vs. 3 mo 34.9 +/- 1.8 kg/m2). At 3 mo the onset of negative alliesthesia, time to abandon experimental session, was shortened (Initial session 33 vs. 3 mo 24 min). The same trend was observed in the time to reach indifference (Initial session 21.9 +/- 3.8 vs. 3 mo 16.2 +/-2.4 min). There was no observed difference in maximum (Initial session +79.5 +/- 11.7; 3 mo +94.5 +/- 9.9 mm) and minimum (Initial session -90.0 +/- 14.4; 3 mo -106 +/- 11.1 mm) hedonic rating. Earlier onset of negative alliesthesia, as seen in our participants, is not consistent with previous hedonic studies that showed delayed or absent negative alliesthesia in participants when below their initial body weight. Therefore, it is hypothesized that the accelerated onset of negative alliesthesia observed in our obese participants after weight loss is suggestive of a lowered body weight set-point. Factors inherent to the weight loss diet studied here, such as mild energetic restriction, lowered palatability, and diet composition, may have played a role in this experimental outcome.


Oral health knowledge attitudes and behaviors of migrant preschooler parents.
The purpose of the study was to establish baseline data about oral health knowledge, attitudes and behaviors of migrant and seasonal farm workers (MSFW). The study focused on MSFWs that are parents of preschool-aged children, and who utilized services at 3 migrant dental clinics. An oral health knowledge attitudes and behaviors survey was developed and pilot tested in 2006. The resulting 34 item survey was administered by trained promotores de salud (community health workers) to 45 parents of preschoolers (15 at each clinic site) served by 3 migrant dental clinics. Parents answered questions as they pertained to their oldest preschooler (up to age 5). Dental visits in the last 12 months were reported for 26 (58%) of the children. Fifteen parents (33%) had a dental visit in the last year. Thirty-five parents (77/8%) reported their child's oral health to be good, and 21 (46.7%) reported their own to be good. Half of the children were enrolled in Head Start (HS). Of those, 18 (79%) had a dental visit in the last year, whereas 8 (36%) of those not enrolled in HS had a visit. Discrepancies existed for the age parents believed children should stop using a bottle and the age they actually did stop using a bottle. There were discrepancies in knowledge about decay causing drinks and consumption of drinks by preschool-aged children. MSFWs remain an underserved population with poor access to oral health care and multiple factors affecting oral health knowledge, attitudes and behaviors. A better understanding of influences on oral health knowledge, attitudes and behaviors within the population can assist in implementing appropriate interventions for the maintenance of good oral health in MSFW families. HS can have a positive impact on oral health for MSFW children.


Proteomic Changes in the Monolayer and Spheroid Melanoma Cell Models of Acquired Resistance to BRAF and MEK1/2 Inhibitors.
Extracellular signal-regulated kinase-1/2 (ERK1/2) pathway inhibitors are important therapies for treating many cancers. However, acquired resistance to most protein kinase inhibitors limits their ability to provide durable responses. Approximately 50% of malignant melanomas contain activating mutations in BRAF, which promotes cancer cell survival through the direct phosphorylation of the mitogen-activated protein kinase MAPK/ERK 1/2 (MEK1/2) and the activation of ERK1/2. Although the combination treatment with BRAF and MEK1/2 inhibitors is a recommended approach to treat melanoma, the development of drug resistance remains a barrier to achieving long-term patient benefits. Few studies have compared the global proteomic changes in BRAF/MEK1/2 inhibitor-resistant melanoma cells under different growth conditions. The current study uses high-resolution label-free mass spectrometry to compare relative protein changes in BRAF/MEK1/2 inhibitor-resistant A375 melanoma cells grown as monolayers or spheroids. While approximately 66% of proteins identified were common in the monolayer and spheroid cultures, only 6.2 or 3.6% of proteins that significantly increased or decreased, respectively, were common between the drug-resistant monolayer and spheroid cells. Drug-resistant monolayers showed upregulation of ERK-independent signaling pathways, whereas drug-resistant spheroids showed primarily elevated catabolic metabolism to support oxidative phosphorylation. These studies highlight the similarities and differences between monolayer and spheroid cell models in identifying actionable targets to overcome drug resistance.