Characteristics of early MRI in children and adolescents with vanishing white matter.
MRI in vanishing white matter typically shows diffuse abnormality of the cerebral white matter, which becomes increasingly rarefied and cystic. We investigated the MRI characteristics preceding this stage. In a retrospective observational study, we evaluated all available MRIs in our database of DNA-confirmed VWM patients and selected MRIs without diffuse cerebral white matter abnormalities and without signs of rarefaction or cystic degeneration in patients below 20 years of age. A previously established scoring list was used to evaluate the MRIs. An MRI of seven patients fulfilled the criteria. All had confluent and symmetrical abnormalities in the periventricular and bordering deep white matter. In young patients, myelination was delayed. The inner rim of the corpus callosum was affected in all patients. In early stages of VWM, MRI does not necessarily display diffuse cerebral white matter involvement and rarefaction or cystic degeneration. If the MRI abnormalities do not meet the criteria for VWM, it helps to look at the corpus callosum. If the inner rim (the callosal-septal interface) is affected, VWM should be considered.


Life review therapy for older adults with depressive symptoms in general practice: results of a pilot evaluation.
General practices play an important role in the detection and treatment of depressive symptoms in older adults. An adapted version of the indicated preventive life review therapy group intervention called Looking for Meaning (LFM) was developed for general practice and a pilot evaluation was conducted. A pretest-posttest design was used. One week before and one week after the intervention participants filled out questionnaires. In six general practices in the Netherlands the adapted intervention was given. Inclusion criteria were > 60 years and a score of 5 or higher on the Center for Epidemiological Studies Depression Scale (CES-D). The length and number of LFM sessions were shortened and the intervention was given by one mental health care nurse practitioner (MHCNP). The impact on mental health was analyzed by depressive symptoms (CES-D) as the primary outcome and anxiety symptoms (HADS-A), psychological well-being (PGCMS) and mastery (PMS) as secondary outcomes. An evaluative questionnaire was included to evaluate the feasibility and acceptability. Most participants were satisfied with the adaptations of the number (72%) and length (72%) of sessions. The overall sample showed a significant decrease in depressive symptoms after the intervention. No impact was found on psychological well-being, anxiety symptoms and mastery. The intervention is feasible and acceptable for older adults with depressive symptoms and has an impact on their depressive symptoms.


Inhibition of S-Adenosylmethionine-Dependent Methyltransferase Attenuates TGFβ1-Induced EMT and Metastasis in Pancreatic Cancer: Putative Roles of miR-663a and miR-4787-5p.
The identification of epigenetic reversal agents for use in combination chemotherapies to treat human pancreatic ductal adenocarcinomas (PDAC) remains an unmet clinical need. Pharmacologic inhibitors of Enhancer of Zeste Homolog 2 (EZH2) are emerging as potential histone methylation reversal agents for the treatment of various solid tumors and leukemia; however, the surprisingly small set of mRNA targets identified with EZH2 knockdown suggests novel mechanisms contribute to their antitumorigenic effects. Here, 3-deazaneplanocin-A (DZNep), an inhibitor of S-adenosyl-L-homocysteine hydrolase and EZH2 histone lysine-N-methyltransferase, significantly reprograms noncoding microRNA (miRNA) expression and dampens TGFβ1-induced epithelial-to-mesenchymal (EMT) signals in pancreatic cancer. In particular, miR-663a and miR-4787-5p were identified as PDAC-downregulated miRNAs that were reactivated by DZNep to directly target TGFβ1 for RNA interference. Lentiviral overexpression of miR-663a and miR-4787-5p reduced TGFβ1 synthesis and secretion in PDAC cells and partially phenocopied DZNep's EMT-resisting effects, whereas locked nucleic acid (LNA) antagomiRNAs counteracted them. DZNep, miR-663a, and miR-4787-5p reduced tumor burden in vivo and metastases in an orthotopic mouse pancreatic tumor model. Taken together, these findings suggest the epigenetic reprogramming of miRNAs by synthetic histone methylation reversal agents as a viable approach to attenuate TGFβ1-induced EMT features in human PDAC and uncover putative miRNA targets involved in the process. The findings support the potential for synthetic histone methylation reversal agents to be included in future epigenetic-chemotherapeutic combination therapies for pancreatic cancer. Mol Cancer Res; 14(11); 1124-35. ©2016 AACR.


Evaluating technology-enhanced learning: A comprehensive framework.
The absence of a standard, comprehensive approach to evaluating technology-enhanced learning (TEL) limits the utility of individual evaluations, and impedes the integration and synthesis of results across studies. To outline a comprehensive framework for approaching TEL evaluation in medical education, and to develop instruments for measuring the perceptions of TEL learners and instructors. Using both theoretical constructs of inquiry in education and a synthesis of existing models and instruments, we outlined a general model for evaluation that links utility, principles, and practices. From this we derived a framework for TEL evaluation that identifies seven data collection activities: needs analysis; documentation of processes, decisions, and final product; usability testing; observation of implementation; assessment of participant experience; assessment of learning outcomes; and evaluation of cost, reusability, and sustainability. We then used existing quality standards and approaches to develop instruments for assessing the experiences of learners and instructors using TEL. No single evaluation is likely to collect all of this information, nor would any single audience likely find all information elements equally useful. However, consistent use of a common evaluation framework across different courses and institutions would avoid duplication of effort and allow cross-course comparisons.


Effect of AEE788 and/or Celecoxib on colon cancer cell morphology using advanced microscopic techniques.
Analysis of changes in cancer cell morphology and cytoskeletal element induced by external stimuli is focus of current cancer chemotherapeutic studies. Cancer cell cytoskeleton is complex network of interwoven protein fibers composed of microtubules, microfilaments and intermediate filaments. These interwoven protein fibers are responsible for maintaining cell morphology, movement, adhesion and transmembrane signal transmission. In this study, morphological and cytoskeletal changes induced by AEE788 and/or Celecoxib on colon cancer cell HCT 15 were analyzed using advanced microscopic techniques. Cell proliferation assay was used for determining IC(50) of AEE788 and/or Celecoxib on HCT 15. Confocal microscopic analysis of AEE788 and/or Celecoxib treated HCT 15 was performed using Rhodamine-Phalloidin (actin stain) and Hoechst 33342 (nuclear stain). Atomic force (AFM) and scanning electron microscopic (SEM) studies were also performed to analyze cell morphology and cell wall extension (filopodia and lamellipodia). In addition, quantitative analysis of morphological parameters was studied using cellular image processing technique. This is the first report that combination of AEE788 and Celecoxib additively increase growth inhibition and cell death on human colon cancer cell HCT 15 as estimated by cell proliferation assay. Morphological analysis of AEE788 or Celecoxib treated HCT 15 cell for 24h have not revealed significant change in morphology under phase contrast microscopy. But, slight morphological changes were observed in combination (AEE788+Celecoxib) treated HCT 15 for 24h. In contrast, high resolution confocal laser fluorescence and atomic force microscopic studies have revealed cell shrinkage, disorganized actin filament and, loss of filopodia and lamellipodia. These changes were more prominent in combination of AEE788 and Celecoxib treated HCT 15 than either drug alone. These results may suggest antiproliferative and antimetastatic activity of AEE788 and/or Celecoxib. Quantitative analysis of morphological parameters using cellular image processing technique have shown decrease in mean area, perimeter, compactness and eccentricity of combination drug treated cells than either drug alone. These results further support the confocal and AFM study. Scanning electron microscopic study of AEE788 and/or Celecoxib treated HCT 15 has also shown morphological changes and loss of filopodia and lamellipodia. In conclusion, this investigation of morphological and cytoskeletal changes using advanced microscopic techniques present a significant foundation for evaluating anticancer activity of a drug and form a new strategy for evaluating effect of AEE788 and/or Celecoxib on colon cancer.


Effect of α-tocopherol tissue levels on beef quality.
To evaluate meat quality of beef with different α-tocopherol tissue levels, 55 feedlot steers were fed a barley-based finisher diet with four vitamin E supplementation levels (0, 350, 700 and 1400 IU DL-α-tocopheryl acetate/animal per day) for 120 days. Although the increase in oxidation levels overtime was much smaller (P < 0.001) in the high-medium and high groups, α-tocopherol tissue levels did not affect (P > 0.05) pH, proximate analysis, drip and cooking losses, and shear force of steaks. No effect of α-tocopherol tissue levels was found in retail evaluation of steaks after a short ageing time of 6 days, but with 21 days of ageing, a delay in formation of metmyoglobin (P = 0.008) was observed in steaks with higher tissue levels of α-tocopherol. Similar results were found for ground beef (25% fat) prepared from 6-day aged meat. Thus, higher α-tocopherol tissue levels protect ground beef and long-aged steaks from discolouration and lipid oxidation.


Quality performance development for education: implementation of a plan.
Measuring or evaluating the impact of educational interventions has presented a challenge for the educator. Implementing a quality performance improvement (QPI) plan addressing education processes and outcomes provides an expansion and extension of QPI, as well as validates the pivotal and evolving role of the educator in measuring patient care outcome. This article describes an education-based QPI plan using the 10-step process identified by the Joint Commission on Accreditation of Healthcare Organizations (JCAHO). A QPI study will clarify application of each step.


The potential and advances in RNAi therapy: chemical and structural modifications of siRNA molecules and use of biocompatible nanocarriers.
Small interfering RNA (siRNA) has attracted great attention as a potential new drug due to its highly sequence-specific gene silencing ability and generality in therapeutic target. However, the medical applications of siRNA have been severely hindered by the lack of an optimal systemic delivery methodology. This poor delivery performance of siRNA is mainly caused by its inherent physicochemical properties including short and stiff structure, low charge density and vulnerability to nuclease cleavage. Thus, the successful development of efficient systemic delivery platform for siRNA is a fundamental requirement necessary to bring siRNA-based drugs to the market. Herein, we describe some siRNA delivery methods based on the chemical and structural modifications of delivery materials and siRNA itself to carry siRNA therapeutics safely to the targeted place without adverse effects. This review particularly explains the latest progress of chemically and structurally modified siRNA polymer (poly-siRNA)-based delivery systems. The stable and compact siRNA polyplexes, which are formed by poly-siRNA and different types of biocompatible materials, can enhance serum stability and target delivery efficiency in vitro and in vivo. In addition, this review provides specific information on poly-siRNA delivery systems from basics to therapeutic applications in different animal disease models.


[Complex correction of functional state in subjects with psychoautonomic syndrome].
The article presents the results of correction of functional condition in patients with vegetative disorders using non-pharmacological factors: modified autogenic training, cyclic physical exercise, respiratory gymnastics, contrast shower. The complex of the non-pharmacological factors corrects defects in interhemispheric relationships, relieves the severity of vegetative affection, improves psychoemotional condition, enhances functional potentialities of cardiovascular and respiratory systems in patients with psychovegetative syndrome.


Decreasing suicide risk among patients with prostate cancer: Implications for depression, erectile dysfunction, and suicidal ideation screening.
Prostate cancer is the most common malignancy among males, accounting for 19% of cancers, and the third most common cancer-related cause of death. Suicide rates in the United States have increased among males over the last decade. Further, suicide rates are higher in oncology patients, including patients with prostate cancer, compared to the general population. The objective of this article is to review the current literature and address the relationship between prostate cancer, depression, erectile dysfunction, and suicidal ideation. We reviewed the current literature pertaining to prostate cancer and depression, and prostate cancer and suicide. Furthermore, associations were made between erectile dysfunction and depression. Men with prostate cancer at increased risk for suicidal death are White, unmarried, elderly, and men with distant disease. Time since diagnosis is also an important factor, since men are at risk of suicide>15 years after diagnosis. Approximately 60% of men with prostate cancer experience mental health distress, with 10%-40% having clinically significant depression. Additionally, patients that received androgen deprivation therapy (ADT) are 23% more likely to develop depression compared to those without ADT. Longitudinal studies of prostate cancer patients suggest that erectile dysfunction after curative treatment may have a significant psychological effect leading to depression. Herein, a newly proposed screening algorithm suggests for an evaluation with the expanded prostate cancer index composite-clinical practice, patient health questionnaire-9, and an 8-question suicidal ideation questionnaire to assess for health-related quality of life, depression, and suicidal ideation. The burden of screening for erectile dysfunction, depression and suicidal ideation lies with the entire health care team, as there appears to be an association between these diagnoses, that is, compounded in patients with prostate cancer. The screening algorithm should assist with guiding timely and appropriate psychiatric referral to optimize outcomes in these high-risk patients.


17β-estradiol and progesterone effect on human papillomavirus 16 positive cells grown as spheroid co-cultures.
Human papillomavirus (HPV) is the key epidemiologic factor of cervical cancer, but additional cofactors are mandatory. Estrogen has been considered as one of those. Here, the aim was to study the effects of steroid hormones on HPV16 E6-E7, estradiol receptors ERα and ERβ, and progesterone receptor (PR) in HPV16-positive cervical carcinoma cell lines SiHa and CaSki grown as epithelial and fibroblast spheroid co-cultures. The spheroid co-cultures were exposured to 17β-estradiol or progesterone from day 7 onwards. mRNA levels of HPV16 E6-E7, ERα, ERβ and PR normalized against GAPDH were analyzed with quantitative reverse transcription-qPCR (RT-qPCR). 17β-estradiol and progesterone decreased HPV16 E6-E7 mRNA expression in CaSki and increased in SiHA co-cultures. In CaSki co-cultures, ERβ expression was blocked after 17β-estradiol exposure while in SiHa cells it slightly increased ERβ expression. PR expression was seen only in CaSki spheroids and it vanished after exposure to steroid hormones. Fibroblasts expressed all three hormone receptors as monolayers but ERβ expression decreased and ERα and PR vanished after co-culturing. Cell culturing platform changes both oncogene and hormone receptor expression in HPV16 positive cervical cancer cell lines. This needs to be considered when in vitro results are extrapolated to in vivo situations.


Immunochemistry of cartilage proteoglycan. Immunodiffusion and gel-electrophoretic studies.
Cartilage proteoglycan is thought to be composed of subunits, core proteins with covalently attached sulphated polysaccharide side chains, which form aggregates by non-covalent association with a link protein. The new technique of non-disruptive extraction followed by fractionation in caesium chloride gradients provides a useful means of preparing relatively pure proteoglycan aggregate, subunit and link fractions. Immunological studies of these fractions led to the identification of an antigen associated with the proteoglycan subunit which was common to several species and to the demonstration of additional species-specific antigens in aggregate and link fractions derived from bovine nasal cartilage. Polyacrylamide-gel electrophoresis with sodium dodecyl sulphate of bovine proteoglycan aggregate and link fractions gave two protein bands in the gels and a protein-polysaccharide band at the origin; subunit fractions gave only the band at the origin. These results are consistent with the current concept of cartilage proteoglycan structure.


Cyclic AMP and c-KIT signaling in familial testicular germ cell tumor predisposition.
Familial testicular germ cell tumors (FTGCTs) are hypothesized to result from the combined interaction of multiple low-penetrance genes. We reported inactivating germline mutations of the cAMP-binding phosphodiesterase 11A (PDE11A) as modifiers of FTGCT risk. Recent genome-wide association studies have identified single-nucleotide polymorphisms in the KITLG gene, the ligand for the cKIT tyrosine kinase receptor, as strong modifiers of susceptibility to both familial and sporadic testicular germ cell tumors. We studied 94 patients with FTGCTs and 50 at-risk male relatives from 63 unrelated kindreds, in whom the PDE11A gene had been sequenced by investigating the association between KITLG genome-wide association study single-nucleotide polymorphisms rs3782179 and rs4474514 and FTGCT risk in these patients and in 692 controls. We also examined cAMP and c-KIT signaling in testicular tissues and cell lines and extended the studies to 2 sporadic cases, one with a PDE11A defect and one without, as a comparison. We found a higher frequency of the KITLG risk alleles in FTGCT patients who also had a PDE11A sequence variant, compared with those with a wild-type PDE11A sequence. In NTERA-2 and Tcam-2 cells transfected with the mutated forms of PDE11A (R52T, F258Y, Y727C, R804H, V820M, R867G, and M878V), cAMP levels were significantly higher, and the relative phosphodiesterase activity was lower than in the wild-type cells. KITLG expression was consistently increased in the presence of PDE11A-inactivating defects, both at the RNA and protein levels, in familial testicular germ cell tumors. The 2 sporadic cases that were studied, one with a PDE11A defect and another without, agreed with the data in FTGTCT and in the cell lines. Patients with FTGCT and PDE11A defects also carry KITLG risk alleles more frequently. There may be an interaction between cAMP and c-KIT signaling in predisposition to testicular germ cell tumors.


Both RadA and RadB are involved in homologous recombination in Pyrococcus furiosus.
RecA and Rad51 proteins are essential for homologous recombination in Bacteria and Eukarya, respectively. Homologous proteins, called RadA, have been described for Archaea. Here we present the characterization of two RecA/Rad51 family proteins, RadA and RadB, from Pyrococcus furiosus. The radA and radB genes were not induced by DNA damage resulting from exposure of the cells to gamma and UV irradiation and heat shock, suggesting that they might be constitutively expressed in this hyperthermophile. RadA had DNA-dependent ATPase, D-loop formation, and strand exchange activities. In contrast, RadB had a very weak ATPase activity that is not stimulated by DNA. This protein had a strong binding affinity for DNA, but little strand exchange activity could be detected. A direct interaction between RadA and RadB was detected by an immunoprecipitation assay. Moreover, RadB, but not RadA, coprecipitated with Hjc, a Holliday junction resolvase found in P. furiosus, in the absence of ATP. This interaction was suppressed in the presence of ATP. The Holliday junction cleavage activity of Hjc was inhibited by RadB in the absence, but not in the presence, of ATP. These results suggest that RadB has important roles in homologous recombination in Archaea and may regulate the cleavage reactions of the branch-structured DNA.


Feeding intervals and fat absorption in preterm babies.
Fat absorption studies were conducted on 18 preterm babies to determine whether continuous nasogastric feeding is as effective as intermittent feeding. The infants ranged from 1 290 g to 1 960 g in birth weight and were assessed at a mean postnatal age of 34 days. Each served as a control by receiving an evaporated milk 1-hourly, 2-hourly, 3-hourly or continuously in random order. Equivalent volumes of milk were ingested over each 3-day period irrespective of the method of feeding and the daily weight gain was similar for the various regimens. Stool transit times did not vary and fat absorption and retention showed no significant differences. Continuous feeding appears to be suitable for preterm babies, especially when there is a need to avoid abdominal distension.


Policies for fostering health science, technology and innovation in Brazil and the role of clinical research.
The purpose of this article is to highlight a number of underlying issues that may be useful for a comprehensive review of the management of Health-Related Science, Technology and Innovation policies (ST&I/H), and its strategies and priorities. It is an analytical study supported by an extensive review of the technical and journalistic literature, clippings, legislation and federal government directives. The results show that the Healthcare Production Complex undeniably and increasingly needs science to maintain itself. One may infer that a framework of institutional milestones is being built in Brazil, to strengthen, guide and encourage Research and Development, and that clinical research creates scientific knowledge to address public healthcare issues by generating new inputs or enhancing existing techniques, processes and technologies that will be produced, marketed and used in the different segments, thus feeding the Healthcare Productive Complex.


Enhanced Fasanella-Servat Procedure for the Graded Repair of Blepharoptosis.
To describe in detail a technique for a modified Fasanella-Servat procedure that provides a simplified method for graded Mueller muscle excision with minimal and controlled tarsal excision. A retrospective study of 71 patients (102 eyelids) who underwent the modified Fasanella-Servat procedure is reported. Measurements include the preoperative, post-phenylephrine, and postoperative margin-to-reflex distance-1, and postoperative upper eyelid height symmetry as determined by the absolute difference between right- and left-sided margin-to-reflex distance-1. Postoperative complications are reported. The average increase in margin-to-reflex distance-1 was 2.4 mm with an average postoperative upper eyelid height symmetry of 0.4 mm. Postoperative asymmetry was 1.5 mm or less in 68 patients, a success rate of 96%. Four patients (6%) exhibited overcorrection, 2 of which required additional surgery. No lagophthalmos or notable eyelid contour abnormalities were seen. The modified Fasanella-Servat technique offers a simple method to isolate and resect Mueller muscle and a minimal segment of tarsus in a quantitative fashion, allowing for a graded repair of blepharoptosis and thereby decreasing the risk of postoperative overcorrection, lagophthalmos, and eyelid contour asymmetry whilst preserving the bulk of tarsus.


Prevalence of intestinal colonization and nosocomial infection with carbapenem-resistant Enterobacteriales in children: a retrospective study.
We investigated the epidemiological surveillance of the intestinal colonization and nosocomial infection of carbapenem-resistant Enterobacteriales (CRE) isolates from inpatients, which can provide the basis for developing effective prevention. A total of 96 CRE strains were collected from 1,487 fecal samples of hospitalized children between January 2016 and June 2017, which were defined as the "CRE colonization" group. In total, 70 CRE clinical isolates were also randomly selected for the comparison analysis and defined as the "CRE infection" group. The antimicrobial susceptibility of all strains was determined by the microdilution broth method. Polymerase chain reaction (PCR) was used to analyze carbapenemase genes, plasmid typing, and integrons. Multilocus sequence typing was further used to determine clonal relatedness. In the "CRE colonization" group, Klebsiella pneumoniae was mostly detected with a rate of 42.7% (41/96), followed by Escherichia coli (34.4%, 33/96) and Enterobacter cloacae (15.6%, 15/96). The ST11 KPC-2 producer, ST8 NDM-5 producer, and ST45 NDM-1 producer were commonly present in carbapenem-resistant K. pneumoniae (CRKPN), carbapenem-resistant E. coli (CRECO), and carbapenem-resistant E. cloacae (CRECL) isolates, respectively. In the "CRE infection" group, 70% (49/70) of strains were K. pneumoniae, with 21.4% E. cloacae (15/70) and 5.7% E. coli (4/70). The ST15 OXA-232 producer and ST48 NDM-5 producer were frequently observed in CRKPN isolates, while the majority of NDM-1-producing CRECL isolates were assigned as ST45. Phylogenetic analysis showed that partial CRE isolates from intestinal colonization and nosocomial infection were closely related, especially for ST11 KPC-2-producing CRKPN and ST45 NDM-1-producing CRECL. Furthermore, plasmid typing demonstrated that IncF and IncFIB were the most prevalent plasmids in KPC-2 producers, while IncX3/IncX2 and ColE were widely spread in NDM producer and OXA-232 producer, respectively. Then, class 1 integron intergrase intI1 was positive in 74.0% (71/96) of the "CRE colonization" group and 52.9% (37/70) of the "CRE infection" group. This study revealed that CRE strains from intestinal colonization and nosocomial infection showed a partial correlation in the prevalence of CRE, especially for ST11 KPC-2-producing CRKPN and ST45 NDM-1-producing CRECL. Therefore, before admission, long-term active screening of rectal colonization of CRE isolates should be emphasized.


Coarctation of the aorta--a theoretical and experimental analysis of the effects of a centrally located arterial stenosis.
Aortic coarctation is a local constriction of the aorta that may severely affect haemodynamics. It is therefore important to quantify these effects. Using Bernoulli's equation and the momentum theorem, the pressure drop is described including the pressure recovery distal to the coarctation and the effects of collateral flow; both laminar and turbulent. Assuming the coarctation and collaterals to be stiff, a quadratic relationship between flow and pressure drop is expected for flow through the coarctation and for turbulent collateral flow. For laminar collateral flow, a linear relationship is expected. The coarctation flow was studied in a model consisting of a rigid tube with local constriction, connected to a flooded-level tank, containing a 36 per cent by weight solution of sucrose, with a viscosity equivalent to that of blood at body temperature. The pressure drop across the constriction showed a quadratic relationship to flow in agreement with theoretical expectations. Pressure recovery in this model was very slight (0-4 mm Hg). Nine patients with aortic coarctation were catheterised. Cardiac output and pressure drop across the coarctation were measured at rest and during supine cycle exercise at two different workloads. The relationship between mean pressure drop and cardiac output tended to be either 'parabolic' or, in some cases, approximately linear, suggesting that the flow situation in aortic coarctation can be quantified by expressions that either linearly or quadratically relate pressure and flow.


Testosterone Replacement Therapy in Deficient Patients With Chronic Heart Failure: A Randomized Double-Blind Controlled Pilot Study.
Testosterone deficiency is associated with heart failure (HF) progression and poor prognosis. Testosterone therapy has been shown to improve exercise capacity in patients with chronic HF, but no trial has evaluated the impact of replacement in patients with demonstrated testosterone deficiency. Prospective, randomized, double-blind, placebo-controlled, and parallel-group trial comparing testosterone replacement with placebo in males with chronic HF with reduced ejection fraction (HFrEF) and testosterone deficiency (NCT01813201). Long-acting undecanoate testosterone at a fixed dose of 1000 mg was supplied by intramuscular injection at inclusion and then every 3 months. The placebo group received isotonic saline serum. Patients were randomly allocated 1:1 to testosterone or placebo while receiving optimal medical therapy, and the study was conducted for 12 months. The final sample comprised 29 patients, 15 in the placebo group and 14 in the testosterone group (aged 65 ± 8, 62% with an ischemic etiology, left ventricular ejection fraction [LVEF] 30% ± 6%, 69% New York Heart Association functional [NYHA II]). After 12 months, testosterone replacement increased testosterone levels ( P = .002) but was not associated with benefit in terms of clinical symptoms and functional capacity including NYHA class, Framingham score, Minnesota Living Heart Failure Questionnaire, 6-minute walk test, or LVEF and N-terminal pro-B-type natriuretic peptide levels. No significant side effects associated with testosterone treatment were observed. No effects were found in other hormonal, metabolic, and bone turnover biomarkers. In patients with HFrEF and testosterone deficiency, replacement therapy was not associated with any significant improvement.


p53 mutations versus EGF receptor expression in giant cell glioblastomas.
Recent studies have shown that there are distinct genetic pathways leading to the most malignant astrocytic neoplasm, the glioblastoma. Primary (de novo) glioblastomas are characterized by amplification/overexpression of the EGF receptor (EGFR) and, less frequently, of the MDM2 gene. Another pathway, operative in the progression of low-grade or anaplastic astrocytomas to secondary glioblastomas, is characterized by the frequent occurrence of p53 mutations. In this study, we assessed p53 mutations and EGFR expression in the giant cell glioblastoma. This rare variant is characterized by unusually large, multinucleated giant cells, but tends to be more confined and has been reported to carry a somewhat more favorable prognosis. We analyzed biopsies from 16 patients (mean age at clinical manifestation, 40 years). DNA sequencing revealed that 12 of 16 (75%) giant cell glioblastomas contained a p53 mutation. In 7 patients with two or more surgical interventions, the p53 mutation was already detected in the first biopsy. Focal EGFR overexpression, including multinucleated giant cells, was observed immunohistochemically in 9 of 16 (56%) tumors. However, most tumor areas lacked immunoreactivity, indicating that EGFR overexpression does not play a significant role in the evolution of this glioblastoma variant. These results suggest that giant cell glioblastomas develop de novo with a short preoperative history (mean, 47 +/- 40 days), but contain genetic alterations similar to those observed in secondary glioblastomas.


[Preventive and therapeutic effects of Yiqi Huayu Recipe on degeneration of articular cartilage in rats with osteoarthritis].
To study the efficacy and possible mechanism of Yiqi Huayu Recipe (YQHYR), a compound traditional Chinese herbal medicine, in preventing and treating degeneration of the articular cartilage in rats with osteoarthritis (OA). A total of 90 one-month-old SD rats were randomly divided into normal control group, untreated group and YQHYR group, with 30 rats in each group. The osteoarthritis was induced by shoulder disarticulation plus upright posture in rats. The rats in YQHYR group were treated with YQHYR at 5-, 7- and 9-month old (4, 6 and 8 months after the surgery) for one month. Ten rats in each subgroup were sacrificed at 6-, 8- and 10-month old (5, 7 and 9 months after the surgery) respectively and the knee joint samples were harvested for detection. Safranin-O/fast green staining was performed to examine the morphology of articular cartilage. The expressions of type II collagen (Col2A1), aggrecan-1 (Agc1), matrix metalloproteinase-13 (MMP-13) and tissue inhibitor of metalloproteinase-1 (TIMP-1) mRNAs were evaluated by real-time fluorescent quantitation polymerase chain reaction. Histological analysis showed that the structure of articular cartilage was seriously destroyed in rats in the untreated group. On the contrary, the degenerative changes of the articular cartilage in the YQHYR group were dropped off. Real-time fluorescent quantitation polymerase chain reaction showed that expressions of Agc1, TIMP-1 and Col2A1 mRNAs were up-regulated in 6- and 10-month-old rats in the YQHYR group as compared with those in the untreated group (P<0.01, P<0.05), but there was no significant difference in expression of MMP-13 mRNA between the YQHYR group and the untreated group. The expression of Agc1 mRNA was up-regulated and the expression of MMP-13 mRNA was down-regulated in the 8-month-old rats in YQHYR group as compared with those in the untreated group (P<0.01). YQHYR can promote the synthesizing of aggrecan and type II collagen in chondrocytes and delay articular cartilage degradation in OA rats.


Chondroitin sulfate N-acetylgalactosaminyltransferase 1 is necessary for normal endochondral ossification and aggrecan metabolism.
Chondroitin sulfate (CS) is a glycosaminoglycan, consisting of repeating disaccharide units of N-acetylgalactosamine and glucuronic acid residues, and plays important roles in development and homeostasis of organs and tissues. Here, we generated and analyzed mice lacking chondroitin sulfate N-acetylgalactosaminyltransferase 1 (CSGalNAcT-1). Csgalnact1(-/-) mice were viable and fertile but exhibited slight dwarfism. Biochemically, the level of CS in Csgalnact1(-/-) cartilage was reduced to ∼50% that of wild-type cartilage, whereas its chain length was similar to wild-type mice, indicating that CSGalNAcT-1 participates in the CS chain initiation as suggested in the previous study (Sakai, K., Kimata, K., Sato, T., Gotoh, M., Narimatsu, H., Shinomiya, K., and Watanabe, H. (2007) J. Biol. Chem. 282, 4152-4161). Histologically, the growth plate of Csgalnact1(-/-) mice contained shorter and slightly disorganized chondrocyte columns with a reduced volume of the extracellular matrix principally in the proliferative layer. Immunohistochemical analysis revealed that the level of both aggrecan and link protein 1 were decreased in Csgalnact1(-/-) cartilage. Western blot analysis demonstrated an increase in processed forms of aggrecan core protein. These results suggest that CSGalNAcT-1 is required for normal levels of CS biosynthesis in cartilage. Our observations suggest that CSGalNAcT-1 is necessary for normal levels of endochondral ossification, and the decrease in CS amount in the growth plate by its absence causes a rapid catabolism of aggrecan.


[Brain monoamine oxidase during aging].
MAO activities were studied in 4 months old and 19 months old Wistar rats. MAO-B activity was higher in brain stem and hemispheres of old rats as compared with that of young animals. MAO-A activity was lowered only in the stem and was not changed in the hemispheres of old rats. Actinomycin D increased MAO-A activity in the brain stem of young rats and lowered MAO-B activity in the hemispheres of old rats. Emotional stress increased MAO-A activity in the stem and hemispheres of both young and old animals. Actinomycin D abolished the stress effect on these two forms of MAO both young and old rats. Delta-sleep including peptide at the dose of 120 micrograms/100 g increased MAO-A activity and lowered MAO-B activity in young animals but did not affect the MAO-A activity in old rats.


Assessment of visceral pain associated with metritis in dairy cows.
Metritis is a common disease in dairy cattle, but to our knowledge, no work has assessed pain associated with this disease. Tissue palpation is commonly used to assess pain in human and veterinary medicine. The objective of this study was to evaluate visceral pain responses during rectal palpation, with and without uterine palpation, in healthy cows and in cows diagnosed with metritis. A total of 49 Holstein dairy cows (mean ± standard deviation parity of 2.8±1.8) were subjected to systematic health checks every 3 d after parturition for 21 d, scoring for vaginal discharge (0 to 4); 13 cows showed a discharge score ≥2 during at least 1 health check and were classified as metritic, whereas 29 cows were classified as healthy and showed no sign of this or any other disease (including mastitis and lameness). Back arch and heart rate variability before examination and during palpation were recorded using video and heart rate monitors. Back arch (cm(2)) on the day of diagnosis was greater in metritic versus healthy cows (1,034±72 vs. 612±48cm(2)), and greater during rectal palpation with uterine palpation versus rectal palpation without uterine palpation (869±45 vs. 777±45cm(2)). Heart rate frequency domain analysis showed that the low-frequency portion was higher in cows with metritis versus healthy cows (16.5±1.2 vs. 12.9±1.0). Time domain analysis showed that the standard deviation between normal to normal interbeat intervals and the root mean square of successive differences both decreased during rectal palpation with uterine palpation versus rectal palpation without uterine palpation (1.9±0.1 vs. 2.5±0.1 and 1.3±0.1 vs. 1.7±0.1, respectively). Together, these results indicate that the inflammation associated with metritis is painful, and that the pain response can be detected during rectal palpation with and without uterine palpation. Rectal palpation with uterine palpation appears to be more aversive than rectal palpation without uterine palpation, suggesting that the former should be avoided when possible.


Production of an anti-Aβ antibody fragment in Pichia pastoris and in vitro and in vivo validation of its therapeutic effect.
ScFv-h3D6 has been shown as an efficient therapy in the 3xTg-AD mouse model of Alzheimer's Disease. Because one of the major bottlenecks for the therapeutic uses of proteins produced in Escherichia coli is their potential contamination with endotoxins, LPS were extensively removed by a rather low-efficient, expensive, and time-consuming purification step. In addition, disulfide scrambling is favored in the reducing bacterial cytoplasm albeit the use of reductase deficient strains. To overcome these hurdles, as well as to improve the yield, the yeast Pichia pastoris, an endotoxin-free host system for recombinant protein production, has been used to produce scFv-h3D6, both in flask and in a fed-batch bioreactor. Comparison of the thermal stability of the obtained protein with that from E. coli showed no differences. Opposite to the case of the protein obtained from E. coli, no disulfide scrambled conformations or LPS traces were detected in that produced in P. pastoris. Cytotoxicity assays in SH-SY5Y neuroblastoma cell-cultures demonstrated that proteins from both expression systems were similarly efficient in precluding Aβ-induced toxicity. Finally, the 3xTg-AD mouse model was used to test the therapeutic effect of both proteins. Quantification of Aβ levels from cortex and hippocampus protein extracts by ELISA, and Aβ-immunohistochemistry, showed that both proteins reduced Aβ burden. This work demonstrates that scFv-h3D6 obtained from P. pastoris shows the same benefits as those already known for that obtained from E. coli, with multiple advantages in terms of recombinant production and safety.


Plasma concentration of visfatin is a new surrogate marker of systemic inflammation in type 2 diabetic patients.
It is not clear whether plasma visfatin level is related with systemic inflammation or diabetic nephropathy in diabetic patients. In this study, we investigated the relationship between plasma visfatin levels and systemic inflammation or diabetic nephropathy in type 2 diabetic patients. In addition, we examined the physiological action of visfatin in cultured adipocytes in diabetic condition. Plasma visfatin concentrations were significantly higher in the diabetic groups than in the controls. Plasma visfatin levels were positively correlated with systolic blood pressure, body weight, fasting blood glucose, plasma levels of MCP-1, urinary albumin excretion (UAE), and carotid intima-media thickness (IMT), and were inversely correlated with plasma adiponectin, and creatinine clearance. However, plasma visfatin concentrations did not show a significant relationship with HbA1C, BMI or HOMA-IR. Regression analysis showed that plasma levels of MCP-1 and UAE were only independent determinants of plasma visfatin concentration. In cultured adipocytes, high glucose and angiotensin II stimuli markedly increased visfatin synthesis. Exogenous visfatin treatment significantly decreased differentiation of adipocytes and increased NF-kappaB transcriptional activity and pro-inflammatory molecules in adipocytes. These findings suggest that visfatin synthesis is activated from adipose tissue in a diabetic environment, induces NF-kappaB activation and leads to activation of pro-inflammatory cytokines and systemic inflammation.


Identification of Five Key Genes Involved in Intrinsic Apoptotic Pathway From Yellow Catfish Pelteobagrus fulvidraco and Their Transcriptional Responses to High Fat Diet (HFD).
The hypothesis of the present study is that apoptosis through an intrinsic mitochondrial pathway may mediate high fat diet (HFD)-induced changes in the metabolism of Pelteobagrus fulvidraco. To this end, we cloned the full-length cDNA sequences of Cycs, Apaf1, Casp9, Casp3a, and Casp3b involved in the mitochondria apoptotic pathway, and explored their mRNA tissue expressions and transcriptional responses to HFD. All of these members shared similar domains to their orthologous vertebrate genes. They were constitutively expressed in all analyzed tissues but varied from tissue to tissue. Compared to the control, HFD up-regulated the mRNA expression of partial genes among these five key genes (Cycs, Apaf1, Casp9, Casp3a, and Casp3b) in mesenteric fat, intestine, ovary and the kidney, indicating the induction of apoptosis in these tissues; in contrast, HFD down-regulated mRNA levels of partial genes among the five key genes (Cycs, Apaf1, Casp9, Casp3a, and Casp3b) in the heart, spleen and gill tissues, indicating the inhibition of apoptosis in these tissues. The present study will facilitate further exploration into the functions of these genes at the molecular level and disclose the critical involvement of these genes against nutrient changes, indicating that processes of apoptosis in various tissues may differentially be modified by HFD.


Fluorescent organo-antimony compounds as precursors for syntheses of redox-active trimeric and dimeric alkali metal antimonides: an insight into electron transfer reduction processes.
(Tip)2SbCl (1, Tip = 2,4,6-triisopropylphenyl) has been utilized as a precursor for the synthesis of the distibane (Tip)4Sb2 (4) via one-electron reduction using KC8. The two-electron reduction of 1 and 4 afforded the novel trinuclear antimonide cluster [K3((Tip)2Sb)3(THF)5] (6). Changing the reducing agent from KC8 to a different alkali metal resulted in the solid-state isolation of corresponding stable dimeric alkali metal antimonides with the general formula [M2((Tip)2Sb)2(THF)p-x(tol)x] (M = Li (14), Na (15), Cs (16)). In this report, different aspects of the various reducing agents [K metal, KC8, and [K2(Naph)2(THF)]] used have been studied, correlating the experimental observations with previous reports. Additional reactivity studies involving 1 and AgNTf2 (Tf = trifluoromethanesulfonyl) afforded the corresponding antimony cation (Tip)2Sb+NTf2- (19). The Lewis acidic character of 19 has been unambiguously proved via treatment with Lewis bases to produce the corresponding adducts 20 and 21. Interestingly, the precursors 1 and 4 have been observed to be highly luminescent, emitting green light under short-wavelength UV radiation. All the reported compounds have been characterized via NMR, UV-vis, mass spectrometry, and single-crystal X-ray diffraction analysis. Cyclic voltammetry (CV) studies of 1 in THF showed possible two electron reduction, suggesting the in situ generation of the corresponding radical-anion intermediate 1˙- and its subsequent conversion to the monomeric intermediate (Tip)2Sb- (5) upon further reduction. 5 undergoes oligomerization in the solid state to produce 6. The existence of 1˙- was proved using electron paramagnetic resonance (EPR) spectroscopy in solution. CV studies of 6 suggested its potential application as a reducing agent, which was further proved via the conversion of Tip-PCl2 to trimeric (Tip)3P3 (17), and cAACP-Cl (cAAC = cyclic alkyl(amino)carbene) to (cAAC)2P2 (18) and 4, utilizing 6 as a stoichiometric reducing agent.


Clinodiplosis agerati (Diptera, Cecidomyiidae), a new galling species associated with Ageratum conyzoides (Asteraceae) from Brazil.
Clinodiplosis agerati, a new galling species that induces stem galls on Ageratum conyzoides (Asteraceae) is described and illustrated (larva, pupa, male, female and gall) based on material collected in Minas Gerais, Brazil. The new species is compared with the other Neotropical species. This is the first record of the Clinodiplosis in Ageratum.


Deep and wide valleys drive nested phylogeographic patterns across a montane bird community.
Montane species distributions interrupted by valleys can lead to range fragmentation, differentiation and ultimately speciation. Paleoclimatic fluctuations may accentuate or reduce such diversification by temporally altering the extent of montane habitat and may affect species differentially. We examined how an entire montane bird community of the Western Ghats--a linear, coastal tropical mountain range--responds to topographic valleys that host different habitats. Using genetic data from 23 species (356 individuals) collected across nine locations, we examined if different species in the community reveal spatial concordance in population differentiation, and whether the timing of these divergences correlate with climatic events. Our results reveal a nested effect of valleys, with several species (10 of 23) demonstrating the oldest divergences associated with the widest and deepest valley in the mountain range, the Palghat Gap. Further, a subset of these 10 species revealed younger divergences across shallower, narrower valleys. We recovered discordant divergence times for all valley-affected montane birds, mostly in the Pleistocene, supporting the Pliestocene-pump hypotheses and highlighting the role of climatic fluctuations during this period in driving species evolution. A majority of species remain unaffected by valleys, perhaps owing to geneflow or extinction-recolonization dynamics. Studying almost the entire community allowed us to uncover a range of species' responses, including some generalizable and other unpredicted patterns.


Polarographic assay and intracellular distribution of superoxide dismutase in rat liver.
1. A polarographic assay of superoxide (O2--) dismutase (EC 220.127.116.11) activity is described, in which the ability of the enzyme to inhibit O2---dependent sulphite oxidation, initiated by xanthine oxidase activity, is measured. The assay was used in a study of the intracellular distribution of superoxide dismutase in rat liver. Both cyanide-sensitive cupro-zinc dismutase (92% of the total activity) and cyanide-insensitive mangano-dismutase (8%) were measured. 2. Rat liver homogenates contained both particulate (16%y and soluble (84%) dismutase activity. The particulate activity contained both types of dismutase, whereas nearly all the soluble dismutase was a cupro-zinc enzymes. The distribution pattern of mangano-dismutase was similar to that of cytochrome oxidase and glutamate dehydrogenase, indicating that the enzyme was probably present exclusively in the mitochondria. 3. Superoxide dismutase activity in the heavy-mitochondrial (M) fraction was latent and was activated severalfold and largely solubilized by sonication. Treatment of the M fraction with digitonin or a hypo-osmotic suspending medium indicated that most of the cupro-zinc dismutase was located in the mitochondrial intermembrane space, whereas the mangano-enzyme was located in the inner-membrane and matrix space. 4. A small amount of dismutase activity appeared to be present in the nuclei and microsomal fraction, but little or no activity in the lysosomes or peroxisomes. 5. The results are discussed in relation to the intracellular location of known O2---generating enzymes, the possible role of superoxide dismutase activity in intracellular H2O2 formation, and to current views on the physiological function of the enzyme.


Impaired mediolateral postural control at the ankle in healthy, middle-aged adults.
Elderly adults sway more than young adults. Based on the literature, the authors expected the mediolateral ankle postural control mechanism to be affected before age 60 years. Twelve healthy young adults (24.21 ± 2.50 years) and 12 middle-aged adults (51.13 ± 6.09 years) participated in the study. To challenge mediolateral stance, the conditions modified the mediolateral distance among the feet (narrow and standard distances), mandibular position (rest position, left and right laterality occlusion positions), and the occlusion with clenching (intercuspal occlusion, left and right maximal voluntary clenches). As we expected, middle-aged adults exhibited significantly reduced contribution of the ankle mechanism. It was so both in narrow and standard stances. A second objective was to show a greater contribution of the 2 mechanisms in narrow than in standard stances. The results confirmed our hypothesis. As we expected, mandibular conditions only had a significant effect on center of pressure sway. Unexpectedly, middle-aged adults did not increase their range of center of pressure sway in narrow stance. They may have overconstrained their center of pressure sway because of their ankle impairments. On the practical level, our results suggest that older adults should increase their stance width to relieve their hip and ankle control mechanisms and to stabilize their mediolateral posture.


[Cytogenetic background of male infertility].
An account is given of cytogenetic causes of male infertility, on the basis of literature and the authors' own observations. Impairment of sexual differentiation was found to be one of the causes of infertility. The functional mechanisms of the Y-chromosome and H-Y antigen are discussed, in that context. The structural changes in the Y-chromosome were found to provide a possibility of gene labelling and to offer an explanation of the genetic processes underlying various types of infertility. An analysis was made also of male fertility accompanied by aberrations of the other chromosomes. The point is made that in the pathogenesis of male infertility decisive importance should be attributed to chromosomal aberrations, above all to deviations of sexual chromosomes. Thorough cytogenetic examination is absolutely essential unless another causative factor has been clearly established.


Murine FATP alleviates growth and biochemical deficiencies of yeast fat1Delta strains.
Saccharomyces cerevisiae is an ideal model eukaryote for studying fatty-acid transport. Yeast are auxotrophic for unsaturated fatty acids when grown under hypoxic conditions or when the fatty-acid synthase inhibitor cerulenin is included in the growth media. The FAT1 gene encodes a protein, Fat1p, which is required for maximal levels of fatty-acid import and has an acyl CoA synthetase activity specific for very-long-chain fatty acids suggesting this protein plays a pivotal role in fatty-acid trafficking. In the present work, we present evidence that Fat1p and the murine fatty-acid transport protein (FATP) are functional homologues. FAT1 is essential for growth under hypoxic conditions and when cerulenin was included in the culture media in the presence or absence of unsaturated fatty acids. FAT1 disruptants (fat1Delta) fail to accumulate the fluorescent long-chain fatty acid fatty-acid analogue 4, 4-difluoro-5-methyl-4-bora-3a,4a-diaza-s-indacene-3-do decanoic acid (C1-BODIPY-C12), have a greatly diminished capacity to transport exogenous long-chain fatty acids, and have very long-chain acyl CoA synthetase activities that were 40% wild-type. The depression in very long-chain acyl CoA synthetase activities were not apparent in cells grown in the presence of oleate. Additionally, beta-oxidation of exogenous long-chain fatty acids is depressed to 30% wild-type levels. The reduction of beta-oxidation was correlated with a depression of intracellular oleoyl CoA levels in the fat1Delta strain following incubation of the cells with exogenous oleate. Expression of either Fat1p or murine FATP from a plasmid in a fat1Delta strain restored these phenotypic and biochemical deficiencies. Fat1p and FATP restored growth of fat1Delta cells in the presence of cerulenin and under hypoxic conditions. Furthermore, fatty-acid transport was restored and was found to be chain length specific: octanoate, a medium-chain fatty acid was transported in a Fat1p- and FATP-independent manner while the long-chain fatty acids myristate, palmitate, and oleate required either Fat1p or FATP for maximal levels of transport. Lignoceryl CoA synthetase activities were restored to wild-type levels in fat1Delta strains expressing either Fat1p or FATP. Fat1p or FATP also restored wild-type levels of beta-oxidation of exogenous long-chain fatty acids. These data show that Fat1p and FATP are functionally equivalent when expressed in yeast and play a central role in fatty-acid trafficking.


Modifiable factors in access to living-donor kidney transplantation among diverse populations.
We have observed a significant inequality in the number of living-donor kidney transplants (LDKT) performed between patients of non-Western European origin and those of Western European origin. The aim of this study was to investigate modifiable factors that could be used as potential targets for an intervention in an attempt to reduce this inequality. A questionnaire on knowledge, risk perception, communication, subjective norm, and willingness to accept LDKT was completed by 160 end-stage renal patients who were referred to the pretransplantation outpatient clinic (participation rate of 92%). The questionnaire was available in nine languages. Multivariate analyses of variance were conducted to explore differences between patients with and without a living donor. There were significantly fewer patients of non-Western descent (11 of 82) that brought a living donor to the outpatient clinic than patients of Western descent (38 of 78). After correcting for the unmodifiable sociodemographic factors non-Western descent, low knowledge, little communication about their kidney disease, and low willingness to communicate with individuals from the social network about LDKT were significantly associated with the absence of a living donor. Knowledge and communication are identified as modifiable factors that are associated with the likelihood of identifying a potential living donor for LDKT. This observation makes knowledge and communication targets for interventions to reduce inequality in access to LDKT.


[Metabolic response of leukocytes as an index of the individual response of an animal to stress and traumatic shock].
A relationship has been established between varying metabolic response of leukocytes to immobilization stress and severity of the shock progress. Initiation of the shock is accompanied by different response of animal leukocytes that correlates with the severity of the subsequent process. Formation of the resistance to shock is associated with an active glucose involvement into energy and plastic metabolic reactions, activation of the substrate-enzymic systems of glycolysis, pentose shunt, terminal stage of the respiratory chain, decreased rate of succinte dehydrogenation.


The excitatory neuronal network of rat layer 4 barrel cortex.
Sensory whiskers are mapped to rodent layer 4 somatosensory cortex as discrete units termed barrels, which can be visualized at high resolution in living brain slices. Both anatomical and physiological properties of the layer 4 neuronal network can thus be investigated in the context of the functional boundaries of this sensory map. Large-scale confinement of neuronal arbors to single barrels was suggested by restricted lateral diffusion of DiI across septa between barrels. Morphological analysis of dendritic and axonal arborizations of individual excitatory neurons showed that neuronal processes remain within the barrel of origin through polarization toward the center of the barrel. Functionally, the large-scale properties of the neuronal network were investigated through mapping the spatial extent of field EPSPs, which were found to attenuate at barrel borders. This ensemble property of a layer 4 barrel was further investigated by analyzing the connectivity of pairs of excitatory neurons with respect to the locations of the somata. Approximately one-third of the excitatory neurons within the same barrel were synaptically coupled. At the septum between adjacent barrels the connectivity dropped rapidly, and very few connections were found between neurons located in adjacent barrels. Each layer 4 barrel is thus composed of an excitatory neuronal network, which to a first order approximation, acts independently of its neighbors.


Combined use of cervical headache snag and cervical snag half rotation techniques in the treatment of cervicogenic headache.
[Purpose] Cervicogenic headache is a major problem in patients with upper cervical dysfunction. However, its physical therapy management is a topic of debate. This study aims to determine the effect of C1-C2 Mulligan sustained natural apophyseal glide mobilizations on cervicogenic headache and associated dizziness. [Participants and Methods] This study included 48 patients with cervicogenic headache, who were randomly assigned to three equal groups: Group A (Headache SNAG), group B (C1-C2 SNAG rotation), and group C (combined). Neck Disability Index was used to examine neck pain intensity and cervicogenic headache symptoms. The 6-item Headache Impact Test scale was used to examine headache severity and its adverse effects on social life and functions. Flexion-Rotation Test was used to assess rotation range of motion at the level of C1-C2 and confirmed by a cervical range of motion device. Dizziness Handicap Inventory scale was used to evaluate dizziness. The evaluation was done pre- and post-treatment and compared between the groups. [Results] Group C showed significant improvement in all variables compared with groups A and B. [Conclusion] Sustained natural apophyseal glide mobilizations used in the study were effective in reducing cervicogenic headache and dizziness in all groups with a greater improvement in the combined group. The use of cervical SNAG mobilizations is encouraged as a noninvasive intervention depending on the therapist's assessment, findings, and clinical reasoning.


Influence of alkyl chain length and anion species on ionic liquid structure at the graphite interface as a function of applied potential.
Atomic force microscopy (AFM) force measurements elucidate the effect of cation alkyl chain length and the anion species on ionic liquid (IL) interfacial structure at highly ordered pyrolytic graphite (HOPG) surfaces as a function of potential. Three ILs are examined: 1-hexyl-3-methylimidazolium tris(pentafluoroethyl)trifluorophosphate ([HMIM] FAP), 1-ethyl-3-methylimidazolium tris(pentafluoroethyl)trifluorophosphate ([EMIM] FAP), and 1-ethyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide ([EMIM] TFSA). The step-wise force-distance profiles indicate the ILs adopt a multilayered morphology near the surface. When the surface is biased positively or negatively versus Pt quasireference electrode, both the number of steps, and the force required to rupture each step increase, indicating stronger interfacial structure. At all potentials, push-through forces for [HMIM] FAP are the highest, because the long alkyl chain results in strong cohesive interactions between cations, leading to well-formed layers that resist the AFM tip. The most layers are observed for [EMIM] FAP, because the C2 chains are relatively rigid and the dimensions of the cation and anion are similar, facilitating neat packing. [EMIM] TFSA has the smallest push-through forces and fewest layers, and thus the weakest interfacial structure. Surface-tip attractive forces are measured for all ILs. At the same potential, the attractions are the strongest for [EMIM] TFSA and the weakest for [HMIM] FAP because the interfacial layers are better formed for the longer alkyl chain cation. This means interfacial forces are stronger, which masks the weak attractive forces.


Impact of Disasters on Older Adult Cancer Outcomes: A Scoping Review.
There is an urgent need to address the growing global cancer burden in the context of complex disaster events, which both disrupt access to oncology care and facilitate carcinogenic exposures. Older adults (65 years and older) are a growing population with multifaceted care needs, making them especially vulnerable to disasters. The objective of this scoping review is to characterize the state of the literature concerning older adult cancer-related outcomes and oncologic care after a disaster event. A search was conducted in PubMed and Web of Science. Following Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines for scoping reviews, articles were extracted and screened for inclusion. Eligible articles were summarized using descriptive and thematic analyses. Thirty-five studies met all criteria for full-text review. The majority focused on technological disasters (60%, n = 21), followed by climate-amplified disasters (28.6%, n = 10) and geophysical disasters (11.4%, n = 4). Thematic analysis classified the current evidence into three major categories: (1) studies concerned with carcinogenic exposure and cancer incidence related to the disaster event, (2) studies examining changes in access to cancer care and cancer treatment disruptions as a result of the disaster event, and (3) studies exploring the psychosocial experiences of patients with cancer affected by a disaster event. Few studies focused on older adults specifically, and most of the current evidence focuses on disasters in the United States or Japan. Older adult cancer outcomes after a disaster event are understudied. Current evidence suggests that disasters worsen cancer-related outcomes among older adults by disrupting continuity of care and access to timely treatment. There is a need for prospective longitudinal studies following older adult populations postdisaster and studies focused on disasters in low- and middle-income country contexts.


Megawatt-scale average-power ultrashort pulses in an enhancement cavity.
We investigate power scaling of ultrashort-pulse enhancement cavities. We propose a model for the sensitivity of a cavity design to thermal deformations of the mirrors due to the high circulating powers. Using this model and optimized cavity mirrors, we demonstrate 400 kW of average power with 250 fs pulses and 670 kW with 10 ps pulses at a central wavelength of 1040 nm and a repetition rate of 250 MHz. These results represent an average power improvement of one order of magnitude compared to state-of-the-art systems with similar pulse durations and will thus benefit numerous applications such as the further scaling of tabletop sources of hard x rays (via Thomson scattering of relativistic electrons) and of soft x rays (via high harmonic generation).


Cutting-edge statistical methods for a life-course approach.
Advances in research methods, data collection and record keeping, and statistical software have substantially increased our ability to conduct rigorous research across the lifespan. In this article, we review a set of cutting-edge statistical methods that life-course researchers can use to rigorously address their research questions. For each technique, we describe the method, highlight the benefits and unique attributes of the strategy, offer a step-by-step guide on how to conduct the analysis, and illustrate the technique using data from the National Institute of Child Health and Human Development Study of Early Child Care and Youth Development. In addition, we recommend a set of technical and empirical readings for each technique. Our goal was not to address a substantive question of interest but instead to provide life-course researchers with a useful reference guide to cutting-edge statistical methods.


The free-energy landscape of clusters of attractive hard spheres.
The study of clusters has provided a tangible link between local geometry and bulk condensed matter, but experiments have not yet systematically explored the thermodynamics of the smallest clusters. Here we present experimental measurements of the structures and free energies of colloidal clusters in which the particles act as hard spheres with short-range attractions. We found that highly symmetric clusters are strongly suppressed by rotational entropy, whereas the most stable clusters have anharmonic vibrational modes or extra bonds. Many of these clusters are subsets of close-packed lattices. As the number of particles increases from 6 to 10, we observe the emergence of a complex free-energy landscape with a small number of ground states and many local minima.


Digital reconstructed radiography with multiple color image overlay for image-guided radiotherapy.
Registration of patient anatomical structures to the reference position is a basic part of the patient set-up procedure. Registration of anatomical structures between the site of beam entrance on the patient surface and the distal target position is particularly important. Here, to improve patient positional accuracy during set-up for particle beam treatment, we propose a new visualization methodology using digitally reconstructed radiographs (DRRs), overlaid DRRs, and evaluation of overlaid DRR images in clinical cases. The overlaid method overlays two DRR images in different colors by dividing the CT image into two CT sections at the distal edge of the target along the treatment beam direction. Since our hospital uses fixed beam ports, the treatment beam angles for this study were set at 0 and 90 degrees. The DRR calculation direction was from the X-ray tube to the imaging device, and set to 180/270 degrees and 135/225 degrees, based on the installation of our X-ray imaging system. Original and overlaid DRRs were calculated using CT data for two patients, one with a parotid gland tumor and the other with prostate cancer. The original and overlaid DRR images were compared. Since the overlaid DRR image was completely separated into two regions when the DRR calculation angle was the same as the treatment beam angle, the overlaid DRR visualization technique was able to provide rich information for aiding recognition of the relationship between anatomical structures and the target position. This method will also be useful in patient set-up procedures for fixed irradiation ports.


Microtubule alignment and manipulation using AC electrokinetics.
The kinesin-microtubule system plays an important role in intracellular transport and is a model system for integrating biomotor-driven transport into microengineered devices. AC electrokinetics provides a novel tool for manipulating and organizing microtubules in solution, enabling new experimental geometries for investigating and controlling the interactions of microtubules and microtubule motors in vitro. By fabricating microelectrodes on glass substrates and generating AC electric fields across solutions of microtubules in low-ionic-strength buffers, bundles of microtubules are collected and aligned and the electrical properties of microtubules in solution are measured. The AC electric fields result in electro-osmotic flow, electrothermal flow, and dielectrophoresis of microtubules, which can be controlled by varying the solution conductivity, AC frequency, and electrode geometry. By mapping the solution conductivity and AC frequency over which positive dielectrophoresis occurs, the apparent conductivity of taxol-stabilized bovine-brain microtubules in PIPES buffer is measured to be 250 mS m(-1). By maximizing dielectrophoretic forces and minimizing electro-osmotic and electrothermal flow, microtubules are assembled into opposed asters. These experiments demonstrate that AC electrokinetics provides a powerful new tool for kinesin-driven transport applications and for investigating the role of microtubule motors in development and maintenance of the mitotic spindle.


Relationships among Different Water-Soluble Choline Compounds Differ between Human Preterm and Donor Milk.
Choline is essential for infant development. Human milk choline is predominately present in three water-soluble choline (WSC) forms: free choline (FC), phosphocholine (PhosC), and glycerophosphocholine (GPC). It is unclear whether mother's own preterm milk and pooled donor milk differ in WSC composition and whether WSC compounds are interrelated. Mother's own preterm milk (n = 75) and donor milk (n = 30) samples from the neonatal intensive care unit, BC Women's Hospital were analyzed for WSC composition using liquid chromatography tandem mass spectrometry (LC-MS/MS). Associations between different WSC compounds were determined using Pearson's correlations, followed by Fischer r-to-z transformation. Total WSC concentration and concentrations of FC, PhosC, and GPC did not significantly differ between mother's own milk and donor milk. FC was negatively associated with PhosC and GPC in mother's own milk (r = -0.27, p = 0.02; r = -0.34, p = 0.003, respectively), but not in donor milk (r = 0.26, p = 0.181 r = 0.37, p = 0.062, respectively). The difference in these associations between the two milk groups were statistically significant (p = 0.03 for the association between PhosC and FC; and p = 0.003 for the association between FC and GPC). PhosC and GPC were positively associated in mother's own milk (r = 0.32, p = 0.036) but not donor milk (r = 0.36, p = 0.062), although the difference in correlation was not statistically significant. The metabolic and clinical implications of these associations on the preterm infant need to be further elucidated.


Current and Projected Burden of Disease From High Ambient Temperature in Korea.
The objective of the present study was to estimate the current and projected burden of disease from high ambient temperature using population-based data sources of nationwide mortality and morbidity in Korea. Disability-adjusted life years (DALY) were estimated using noninjury-related deaths, and cerebrovascular and cardiovascular diseases from recently released nationwide health and mortality databases. Years of life lost and years lost due to disability were measured based on the point prevalence and number of deaths during the study period. Future DALY attributable to heat waves were estimated from projected populations, and temperature predictions for the years 2030 and 2050 were under Representative Concentration Pathways (RCP) 4.5 and 8.5 with summertime temperatures above threshold. Relative risks (RR) of total mortality and of cardiovascular disease were 1.02 (95% CI, 1.01, 1.02) and 1.08 (95% CI, 1.06, 1.09) for each 1°C increase in temperature above threshold, respectively. The morbidity of heat-related disease was RR 1.67 (95% CI, 1.64, 1.68) for each 1°C increase in temperature above threshold. DALY for all-cause death were 0.49 DALY/1000 in 2011, 0.71 (0.71) DALY/1000 in 2030 and 0.77 (1.72) DALY/1000 in 2050 based on RCP 4.5 (RCP 8.5). DALY for cardio- and cerebrovascular diseases were 1.24 DALY/1000 in 2011, 1.63 (1.82) DALY/1000 in 2030, and 1.76 (3.66) DALY/1000 in 2050 based on RCP 4.5 (RCP 8.5). Future excess mortality due to high ambient temperature is expected to be profound in Korea. Efforts to mitigate climate change can provide substantial health benefits via reducing heat-related mortality.


Michigan assessment-screening test for alcohol and drugs (MAST/AD): evaluation in a clinical sample.
In this study, we sought to evaluate a modification of the Michigan Alcohol Screening Test designed to include problems associated with other drug abuse/dependence besides alcohol. Scores of the lifetime Michigan Assessment-Screening Test/Alcohol-Drug (MAST/AD) were compared to other lifetime measures of substance abuse and dependence and to psychiatric scales reflecting current or recent symptoms. Two university medical centers with alcohol-drug programs located within departments of psychiatry hosted 520 patients with alcohol-drug-related diagnoses. Patients completed their own MAST/AD using a paper-and-pencil format. Based on interviews with the patient, a research associate rated the patients' substance-related problems on the Minnesota Substance Abuse Problem Scale (M-SAPS) and obtained information on lifetime treatment for substance abuse. An addictions psychiatrist determined abuse or dependence and made a current diagnosis of alcohol abuse/dependence only, drug abuse/dependence only, and alcohol plus drug abuse/dependence. The MAST/AD was highly correlated with the M-SAPS and several other measures of substance abuse morbidity. Patients with alcohol-only and drug-only diagnoses did not differ from one another on the MAST/AD, although both groups had lower scores than those with alcohol plus drug diagnoses. Current psychosocial morbidity as assessed by the patient and the psychiatrist was associated with the MAST/AD, although less strongly than with lifetime substance abuse measures. The lifetime MAST/AD demonstrates reliability as a severity measure for alcohol and/or drug abuse. With minor modification, this standard measure can be expanded from alcohol diagnoses to all substance diagnoses. This study in a clinical population did not demonstrate its utility as a screening instrument; additional work is needed to reveal its utility for this purpose.


Three rare diseases in one Sib pair: RAI1, PCK1, GRIN2B mutations associated with Smith-Magenis Syndrome, cytosolic PEPCK deficiency and NMDA receptor glutamate insensitivity.
The National Institutes of Health Undiagnosed Diseases Program evaluates patients for whom no diagnosis has been discovered despite a comprehensive diagnostic workup. Failure to diagnose a condition may arise from the mutation of genes previously unassociated with disease. However, we hypothesized that this could also co-occur with multiple genetic disorders. Demonstrating a complex syndrome caused by multiple disorders, we report two siblings manifesting both similar and disparate signs and symptoms. They shared a history of episodes of hypoglycemia and lactic acidosis, but had differing exam findings and developmental courses. Clinical acumen and exome sequencing combined with biochemical and functional studies identified three genetic conditions. One sibling had Smith-Magenis Syndrome and a nonsense mutation in the RAI1 gene. The second sibling had a de novo mutation in GRIN2B, which resulted in markedly reduced glutamate potency of the encoded receptor. Both siblings had a protein-destabilizing homozygous mutation in PCK1, which encodes the cytosolic isoform of phosphoenolpyruvate carboxykinase (PEPCK-C). In summary, we present the first clinically-characterized mutation of PCK1 and demonstrate that complex medical disorders can represent the co-occurrence of multiple diseases.


Sex-Mediated Differences in LPS Induced Alterations of TNFα, IL-10 Expression, and Prostaglandin Synthesis in Primary Astrocytes.
Although many neurological and psychiatric disorders reveal clear sex-dependent variations, the molecular mechanism of this process is not clear enough. Astrocytes are involved in the response of neural tissue to injury and inflammation, produce steroid hormones, and sense steroid presence. To explore the hypothesis that astrocytes may participate in sex-mediated differences of inflammatory responses, we have examined whether male and female primary rat astrocytes show different responses to lipopolysaccharide (LPS) as a toll-like receptor 4 (TLR4) agonist. Levels of mRNA and proteins of tumor necrosis factor alpha (TNFα), interleukin-10 (IL-10), and cyclooxygenase (COX)-2 were assessed using qPCR, immunoblotting, and ELISA. UPLC-MS/MS was used to detect prostaglandins (PGs). LPS stimulation resulted in different levels of cytokine production; more TNFα and less IL-10 were produced in female cells compared with male astrocytes. Although the levels of the COX-2 expression were not altered, LPS significantly induced the synthesis of PGs with notable sex-related differences. PGE₂ and PGD₂ were less and 6-keto-PGF1α was more upregulated in female astrocytes, and TXB₂ had similar levels in cells obtained from males and females. Trilostane, an inhibitor of 3β-Hydroxysteroid dehydrogenase (3β-HSD), inhibited the LPS-induced TNFα production and the release of PGE₂, PGD₂, and 6-keto-PGF1α in female astrocytes. Thus, male and female astrocytes differentially respond to inflammatory challenges on the level of production of cytokines and steroid hormones. Sex-mediated differences in pro- and anti-inflammatory responses should be taken into consideration for the effective treatment of disorders with neuroinflammation.


Association of prenatal exposure to perfluoroalkyl substances with cord blood adipokines and birth size: The Hokkaido Study on environment and children's health.
Perfluoroalkyl substances (PFASs) are synthetic chemicals that persist in the environment and in humans. There is a possible association between prenatal PFASs exposure and both neonate adipokines and birth size, yet epidemiological studies are very limited. The objective of this study was to examine associations of prenatal exposure to PFASs with cord blood adipokines and birth size. We conducted birth cohort study, the Hokkaido Study. In this study, 168 mother-child pairs were included. Perfluorooctane sulfonate (PFOS) and perfluorooctanoate (PFOA) in maternal blood were determined by liquid chromatography tandem mass spectrometry. Cord blood adiponectin and leptin levels were measured by ELISA and RIA, respectively. Birth weight and ponderal index (PI) were obtained from birth record. The median maternal PFOS and PFOA were 5.1 and 1.4ng/mL, respectively. The median total adiponectin and leptin levels were 19.4μg/mL and 6.2ng/mL, respectively. Adjusted linear regression analyses found that PFOS level was positively associated with total adiponectin levels (β=0.12, 95% CI:0.01, 0.22), contrary was negatively associated with PI (β=-2.25, 95% CI: -4.01, -0.50). PFOA level was negatively associated with birth weight (β=-197, 95% CI: -391, -3). Leptin levels were not associated with PFASs levels. PFOS and adiponectin levels showed marginal dose-response relationship and both PFOS and PFOA and birth size showed significant dose-response relationships. Results from this study suggested that prenatal PFASs exposure may alter cord blood adiponectin levels and may decrease birth size.


Plasma Malondialdehyde (MDA): An Indication of Liver Damage in Women with Pre-Eclamsia.
One of the features associated with pre-eclampsia is elevated liver transaminases. The reason this happens has not been fully described. However, the hepatocytes are not known to be spared by free radical injury. This study was conducted to examine the relationship between product of free radical injury (malondialdehyde) and transaminases in pre-eclamptic women. A total of 98 and 115 pre-eclamptic and apparently normal pregnant women were selected from the booking clinic of Ladoke Akintola University of Technology Teaching Hospital, Osogbo. Non-pregnant women were selected from volunteered members of staff. Malondialdehyde (MDA), aspartate transaminase (AST) and alanine transaminase (ALT) analyses were determined on collected venous blood sample. Statistical analyses of variables were done using SPSS 17 taking level of significance to be p<0.05. Subjects with plasma AST between 10 and 20U/L had mean plasma MDA of 0.92µmol/l whereas those with plasma levels greater than 41U/L had mean plasma MDA of 4.72µmol/l. Similarly, Subjects with plasma ALT between 10 and 20U/L had mean plasma MDA of 0.86µmol/l, and subjects with plasma ALT greater than 51 U/L had mean plasma MDA of 4.71µmol/l. Positive correlation was observed between AST and ALT(r=0.79; p=0.047), between AST and MDA(r=0.690; p=0.061) as well as between ALT and MDA(r=0.571; p=0.049). The elevated liver enzymes seen in women with pre-eclampsia may be due to free radical injury to the liver. Pre-eclamptic women without free radical injury did not have elevated transaminases.


Propensity for Risk in Reproductive Strategy Affects Susceptibility to Anthropogenic Disturbance.
AbstractAnimals initiate, interrupt, or invest resources in reproduction in light of their physiology and the environment. The energetic risks entailed in an individual's reproductive strategy can influence the ability to cope with additional stressors, such as anthropogenic climate change and disturbance. To explore the trade-offs between internal state, external resource availability, and reproduction, we applied state-dependent life-history theory (SDLHT) to a dynamic energy budget (DEB) model for long-finned pilot whales (Globicephala melas). We investigated the reproductive strategies emerging from the interplay between fitness maximization and propensity to take energetic risks, as well as the resulting susceptibility of individual vital rates to disturbance. Without disturbance, facultative reproductive behavior from SDLHT and fixed rules in the DEB model led to comparable individual fitness. However, under disturbance, the reproductive strategies emerging from SDLHT increased vulnerability to energetic risks, resulting in lower fitness than fixed rules. These fragile strategies might therefore be unlikely to evolve in the first place. Heterogeneous resource availability favored more cautious (and thus more robust) strategies, particularly when knowledge of resource variation was accurate. Our results demonstrate that the assumptions regarding the dynamic trade-offs underlying an individual's decision-making can have important consequences for predicting the effects of anthropogenic stressors on wildlife populations.


Molecular cloning and primary structure of rat testes metalloendopeptidase EC 188.8.131.52.
The complete amino acid sequence of rat testes metalloendopeptidase (EC 22.214.171.124) was deduced from the nucleotide sequence of a cDNA clone isolated by screening a rat testes library with a polyclonal antibody raised against a homogeneous preparation of the rat testes enzyme. The correctness of the sequence was verified by N-terminal amino acid sequence analysis of the isolated enzyme and by partial amino acid sequence analysis of three tryptic peptides located near the N-terminus, the middle, and C-terminus of the native protein. The enzyme is composed of 645 amino acids with a molecular weight of 72,985. This value is close to that of the purified rat testes and brain enzyme as determined by polyacrylamide gel electrophoresis under denaturing and reducing conditions and by molecular sieving chromatography. The enzyme contains the putative active-site sequence -H-E-F-G-H- that is homologous to the sequence in the active site of thermolysin and several other related bacterial enzymes, as well as to active-site sequences of several mammalian zinc metallopeptidases. No amino acid sequence homology, beyond this active site, was found with thermolysin, a bacterial zinc metalloendopeptidase, nor with several mammalian zinc metallopeptidases. Northern blot hybridization analyses showed the presence of mRNA encoding the enzyme in rat testes, but not in other rat tissues in spite of the finding that enzyme activity is widely distributed in all tissues and that relatively high activities are present in rat brain and pituitary.


Is there an alternative to the proteasome in cytosolic protein degradation?
While it is clear that the proteasome is the major player in degradative proteolysis in the nucleus and cytosol, there is a lack of complete agreement on whether there are alternative proteolytic pathways or activities responsible for a significant degradation of cytosolic/nuclear substrates. Particularly relevant is the case of the aminopeptidase TPPII (tripeptidyl peptidase II), which has been suggested to be able to perform some of the proteasome functions. However, the current evidence seems to support only a limited role for these cytosolic alternatives. On the other hand, there is evidence of an alternative, autophagy, a pathway involving the delivery of cytosolic substrates to the lysosome for degradation.


Recent advances in preventing stroke recurrence.
Recent advances in secondary stroke prevention include new evidence in hypertension, nutrition, anticoagulation, antiplatelet therapy, intracranial stenosis, percutaneous closure of patent foramen ovale, and lipid-lowering therapy. Individualized therapy for hypertension based on phenotyping with plasma renin and aldosterone markedly improves blood pressure control in patients with resistant hypertension. A Mediterranean diet can reduce the risk of stroke by nearly half. The diagnosis and treatment of metabolic vitamin B12 deficiency, and B vitamins to lower homocysteine, can reduce the risk of stroke by approximately 30%. There are problems with clopidogrel that can be overcome by using ticagrelor, and new anticoagulant drugs markedly improve anticoagulation for stroke prevention, particularly in atrial fibrillation. There are pharmacokinetic problems with dabigatran that deserve attention. Intensive medical therapy is better than stenting for intracranial stenosis, and new therapies directed at proprotein convertase subtilisin-kexin type 9 (PCSK9) will revolutionize lipid-lowering therapy. In the past, it was estimated that rational therapy could reduce recurrent stroke by about 80%. With recent advances, we should be able to do even better.


Acute and chronic effects of neurotrophic factors BDNF and GDNF on responses mediated by thermo-sensitive TRP channels in cultured rat dorsal root ganglion neurons.
Neurotrophic factors (NTFs), beside regulating neuronal survival in the central and peripheral nervous system, are also involved in the modulation of neuronal function in the adult animal. Both brain-derived neurotrophic factor (BDNF) and glial cell-derived neurotrophic factor (GDNF) levels are altered in pathological pain states, and exogenous BDNF and GDNF have multiple effects on pain behavior, depending on the animal model (i.e. inflammatory vs. neuropathic). Thermally gated TRP channels TRPM8, TRPA1 and TRPV1 play a significant role in pain signaling and their pattern and level of expression as well as their biophysical properties are altered in chronic pain states. Our aim was to investigate the effect of long-term and acute exposure to BDNF and GDNF on the functional expression of these thermoTRP channels in cultured rat dorsal root ganglion (DRG) neurons. We found that while BDNF treatment primarily increased the fraction of capsaicin-sensitive (TRPV1-expressing) neurons, GDNF exposure led to an increase in the allyl isothiocyanate (AITC)-responding (TRPA1-expressing) population. Moreover, BDNF treatment increased the amplitude of the response to both AITC and capsaicin. Acute treatment with both NTFs leads to a reduction in the magnitude of tachyphylaxis to noxious stimuli (heat and AITC). Overall, our data provides evidence for a role of BDNF and GDNF in regulating the pattern of expression and level of activity of the transducer channels TRPA1 and TRPV1, leading to enhanced neuronal sensitivity to painful stimuli and increased co-expression of thermoTRP channels.


Identification of the pr1 gene product completes the anthocyanin biosynthesis pathway of maize.
In maize, mutations in the pr1 locus lead to the accumulation of pelargonidin (red) rather than cyanidin (purple) pigments in aleurone cells where the anthocyanin biosynthetic pathway is active. We characterized pr1 mutation and isolated a putative F3'H encoding gene (Zmf3'h1) and showed by segregation analysis that the red kernel phenotype is linked to this gene. Genetic mapping using SNP markers confirms its position on chromosome 5L. Furthermore, genetic complementation experiments using a CaMV 35S::ZmF3'H1 promoter-gene construct established that the encoded protein product was sufficient to perform a 3'-hydroxylation reaction. The Zmf3'h1-specific transcripts were detected in floral and vegetative tissues of Pr1 plants and were absent in pr1. Four pr1 alleles were characterized: two carry a 24 TA dinucleotide repeat insertion in the 5'-upstream promoter region, a third has a 17-bp deletion near the TATA box, and a fourth contains a Ds insertion in exon1. Genetic and transcription assays demonstrated that the pr1 gene is under the regulatory control of anthocyanin transcription factors red1 and colorless1. The cloning and characterization of pr1 completes the molecular identification of all genes encoding structural enzymes of the anthocyanin pathway of maize.


The age of a neutral mutant persisting in a finite population.
Formulae for the mean and the mean square age of a neutral allele which is segregating with frequency x in a population of effective size N(e) have been obtained using the diffusion equation method, for the case of 4N(e)v<1 where v is the mutation rate. It has been shown that the average ages of neutral alleles, even if their frequencies are relatively low, are quite old. For example, a neutral mutant whose current frequency is 10% has the expected age roughly equal to the effective population size N(e) and the standard deviation 1.4N(e) (in generations), assuming that this mutant has increased by random drift from a very low frequency. Also, formulae for the mean "first arrival time" of a neutral mutant to a certain frequency x have been presented. In addition, a new, approximate method has been developed which enables us to obtain the condition under which frequencies of "rare" polymorphic alleles among local populations are expected to be uniform if the alleles are selectively neutral.-It was concluded that exchange of only a few individuals on the average between adjacent colonies per generation is enough to bring about such a uniformity of frequencies.


Novel combination of sorafenib and celecoxib provides synergistic anti-proliferative and pro-apoptotic effects in human liver cancer cells.
Molecular targeted therapy has shown promise as a treatment for advanced hepatocellular carcinoma (HCC). Sorafenib, a multikinase inhibitor, recently received FDA approval for the treatment of advanced HCC. However, although sorafenib is well tolerated, concern for its safety has been expressed. Celecoxib (Celebrex®) is a selective cyclooxygenase-2 (COX-2) inhibitor which exhibits antitumor effects in human HCC cells. The present study examined the interaction between celecoxib and sorafenib in two human liver tumor cell lines HepG2 and Huh7. Our data showed that each inhibitor alone reduced cell growth and the combination of celecoxib with sorafenib synergistically inhibited cell growth and increased apoptosis. To better understand the molecular mechanisms underlying the synergistic antitumor activity of the combination, we investigated the expression profile of the combination-treated liver cancer cell lines using microarray analysis. Combination treatment significantly altered expression levels of 1,986 and 2,483 transcripts in HepG2 and Huh7 cells, respectively. Genes functionally involved in cell death, signal transduction and regulation of transcription were predominantly up-regulated, while genes implicated in metabolism, cell-cycle control and DNA replication and repair were mainly down-regulated upon treatment. However, combination-treated HCC cell lines displayed specificity in the expression and activity of crucial factors involved in hepatocarcinogenesis. The altered expression of some of these genes was confirmed by semi-quantitative and quantitative RT-PCR and by Western blotting. Many novel genes emerged from our transcriptomic analyses, and further functional analyses may determine whether these genes can serve as potential molecular targets for more effective anti-HCC strategies.


[Laparoscopy in urology].
Since the past 10 years, results have established laparoscopy's efficacy. It is actually a consistent surgical option for a lot of indications met in urology. The rational behind performing laparoscopic procedures includes shorter hospital stays, less postoperative pain and a more rapid return to usual activity. Drawbacks of laparoscopy include significant learning curve, longer operative times and higher overall costs. One particular focus is the oncologic applications of laparoscopy for nephrectomy and specially for radical prostatectomy. Laparoscopy become nowadays an usual part of the armamenturium of urological teams.


A NiII-cluster-based MOF as an efficient heterogeneous catalyst for the chemical transformation of CO2.
A novel 3D metal-organic framework based on Ni4 clusters as nodes has been synthesized using a tetrahedral linker of 5,5',5''-(methylsilanetriyl)triisophthalic acid (H6L). The compound possesses 1D channels with potential pore volumes that can accommodate guest molecules. Investigations on the coupling reaction of epoxides and CO2 indicate that the compound can be used as an efficient catalyst for CO2 chemical fixation, and its catalytic performance is greatly superior to most MOF-based catalysts. Additionally, compound 1 can be recycled at least five times without compromising its activity.


Study on anti-Ehrlich ascites tumour effect of Pinellia ternata polysaccharide in vivo.
The objectives of the study were to investigate the anti-tumour activity of Pinellia ternata polysaccharide in vivo, and to preliminarily explore the possible mechanism of its antitumour action. Mouse model of Ehrlich ascites tumour (solid tumour) was used to detect the serum SOD, MDA and GSH-Px levels in mouse and to measure the tumour inhibition rate and survival prolongation rate. The results showed that Pinellia ternata polysaccharide had some tumour inhibitory effect. Tumour weight of Pinellia ternata polysaccharide high-dose group was highly significantly different (P<0.01) compared with the model group. Tumour weight between Pinellia ternata polysaccharide medium-dose group and model group also had a significant difference (P<0.05). Moreover, in the Pinellia ternata polysaccharide high-dose group, survival prolongation rate of ascites tumour mice reached 62.23%, and mouse serum SOD, MDA and GSH-Px levels also rose in varying degrees. The study concluded that Pinellia ternata polysaccharide extract had some in vivo anti-tumour effects, which were probably associated with the enhancement of the body's ability to scavenge excess free radicals by improving the body's enzyme activity.


Traumatic Cervical Vertical Translational Injury: A Case Report.
Traumatic cervical translational injury is a notably rare and highly unstable subtype of type C sub-axial cervical spine injury with high morbidity and mortality rates. Hereby, we report a 41-years-old man who was a case of multiple trauma due to car rollover. He was completely conscious, complaining of cervical pain, with a GCS score of 15/15. His neurological examination was unremarkable. The cervical CT scan revealed a vertical translation at the C6-C7 level (roughly 11 mm) and bilateral facet joint diastases which are highly unstable injuries. A two-stage combined anterior and posterior fixation operation was performed. First, an anterior cervical discectomy and fusion with autologous graft and plate fixation, and then a posterior approach with lateral mass screw fixation was performed. Disruption of the anterior longitudinal ligament, annulus fibrosis, facet capsules, and severe strain of ligamentum flavum was noted intraoperatively. He had no early and late complications within 2 years of follow-up.


Jannaschia rubra sp. nov., a red-pigmented bacterium isolated from sea water.
A Gram-negative, slightly halophilic, strictly aerobic, chemo-organotrophic bacterium was isolated from Mediterranean sea water near Valencia (Spain). Comparison of the almost complete 16S rRNA gene sequence showed that strain 4SM3(T) belonged to the Roseobacter group, with Jannaschia helgolandensis as its closest relative, with a similarity of 98.7 %. DNA-DNA hybridization analysis showed that the Mediterranean isolate had a level of relatedness of less than 42 % with J. helgolandensis and therefore that it represented a novel species of the genus Jannaschia. Phenotypic characteristics gave further evidence that the two organisms are not related at the species level. Isolate 4SM3(T) grows on solid media as irregular pink-red colonies that penetrate into the agar. Cells are rods, motile by a tuft of polar flagella. The DNA base composition is 64.6 mol% G+C. Morphological, physiological and genotypic differences from related species support the description of a novel species, Jannaschia rubra sp. nov., with strain 4SM3(T) (=CECT 5088(T)=DSM 16279(T)) as the type strain.