The place of knowledge management in influencing lasting health change in Africa: an analysis of AMREF's progress.
AMREF (African Medical and Research Foundation) developed a Knowledge Management Strategy that focused on creating, capturing and applying health knowledge to close the gap between communities and health systems in Africa. There was need to identify AMREF's current Knowledge Management implementation status, problems and constraints encountered after two years of enforcement of the strategy and suggest the way forward. This study was conducted between October 2011 and February 2012. Quantitative data on number and foci of AMREF research publications were collected using a questionnaire. Focus group discussions and in-depth interviews were used to gather data on explanations for the trend of publications and the status of the implementation of the 2010-2014 Knowledge Management Strategy. Quantitative data was analysed using SPSS computer software whereas content analysis of themes was employed on qualitative data. Between 1960 and 2011, AMREF produced 257 peer reviewed publications, 158 books and manuals and about 1,188 technical publications including evaluations, guidelines and technical reports. However, the numbers of publications declined from around the year 2000. Large quantities of unpublished and unclassified materials are also in the custody of Heritage. Barriers to Knowledge Management included: lack of incentives for documentation and dissemination; limited documentation and use of good practices in programming; and superficial attention to results or use of evidence. Alternative ways of reorganizing Knowledge Management will enable AMREF to use evidence-based knowledge to advocate for appropriate changes in African health policies and practices.


Angiogenic markers, neovascularization and malignant deformation of Barrett's esophagus.
The molecular events underlying progression of Barrett's esophagus to adenocarcinoma remain an area of active investigation. Neovascularization and angiogenesis have been studied in esophageal adenocarcinomas by counting of microvessels after staining with vascular markers, and by immunohistochemistry for vascular endothelial growth factor. Angiogenesis appears to be increased early in the neoplastic process, but has poor prognostic value. We have demonstrated that expression levels of two important genes that regulate cell growth, namely inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2, are frequently upregulated in Barrett's esophagus and associated adenocarcinomas. COX-2 expression may be related to reflux of bile salts, which induce COX-2 expression in Barrett's tissues and esophageal adenocarcinoma cells in vitro. COX-2 inhibition induces apoptosis and blocks proliferation in COX-2-expressing esophageal adenocarcinoma cells in vitro, and blocks angiogenesis in both in vivo and in vitro models. Although controversial, recent evidence suggest that iNOS-derived NO can inhibit angiogenesis in some model systems. In conclusion, both iNOS and COX-2 appear to be involved in Barrett's-associated neoplastic progression, but COX-2 inhibition is more promising as a chemopreventive strategy. COX-2 inhibition may exert beneficial effects by decreasing angiogenesis and epithelial proliferation, and by facilitating apoptosis of epithelial cells that have undergone DNA damage.


The piRNA pathway in planarian flatworms: new model, new insights.
PIWI-interacting RNAs (piRNAs) are small regulatory RNAs that associate with members of the PIWI clade of the Argonaute superfamily of proteins. piRNAs are predominantly found in animal gonads. There they silence transposable elements (TEs), regulate gene expression and participate in DNA methylation, thus orchestrating proper germline development. Furthermore, PIWI proteins are also indispensable for the maintenance and differentiation capabilities of pluripotent stem cells in free-living invertebrate species with regenerative potential. Thus, PIWI proteins and piRNAs seem to constitute an essential molecular feature of somatic pluripotent stem cells and the germline. In keeping with this hypothesis, both PIWI proteins and piRNAs are enriched in neoblasts, the adult stem cells of planarian flatworms, and their presence is a prerequisite for the proper regeneration and perpetual tissue homeostasis of these animals. The piRNA pathway is required to maintain the unique biology of planarians because, in analogy to the animal germline, planarian piRNAs silence TEs and ensure stable genome inheritance. Moreover, planarian piRNAs also contribute to the degradation of numerous protein-coding transcripts, a function that may be critical for neoblast differentiation. This review gives an overview of the planarian piRNA pathway and of its crucial function in neoblast biology.


Neutrophil-to-lymphocyte Ratio and Its Relation with Markers of Inflammation and Myocardial Necrosis in Patients with Acute Coronary Syndrome.
Inflammation plays an important role in atherosclerosis which is the primary cause of acute coronary syndrome (ACS) that encompasses acute myocardial infarction (AMI) and unstable angina (UA). To investigate and characterize white blood cells (WBC) count, differential blood count in peripheral blood and neutrophil to lymphocyte ratio (NLR) in patients by the type of ACS. The cross-sectional study included 100 patients with ACS (50 males, 50 females), aged 41 to 91 years, classified into two groups: AMI group (n=50) and UA group (n=50). Patients were hospitalized at the Clinic for Heart Diseases, University Clinical Center of Sarajevo. From patients' medical histories the following data were obtained: WBC, neutrophil, eosinophil and basophil granulocytes count, monocyte and lymphocyte count, levels of high sensitive troponin I (hsTnI), creatine kinase MB (CK-MB) and C-reactive protein (CRP). The results were analyzed using software package SPSS, version 19.0. Average WBC count, neutrophil granulocytes, and monocytes were significantly higher in AMI group than in UA group (p = 0.001, p < 0.0005, p = 0.03, respectively). Eosinophil count was significantly lower in patients with AMI (p = 0.022). NLR was significantly higher in AMI group in relation to patients with UA (p = 0.001). Significantly higher values of hsTnI and CK-MB were established in patients with AMI. NLR correlated significantly positive with the values of hsTnI, CK-MB, CRP, WBC and neutrophil count, and significantly negative with lymphocyte count. Average values of NLR were significantly higher in patients with AMI in relation to patients with UA, indicating the importance of this inflammatory marker in discrimination of clinical forms of ACS. A positive correlation was established between NLR and markers of myocardial necrosis, and between NLR and CRP, indicating the importance of NLR in the assessment of the extent of the myocardial lesion and in inflammation intensity assessment in ACS.


Anesthesia for pediatric ophthalmologic surgery.
This review presents updated recommendations, based on existing clinical research, for anesthetic management of strabismus surgery in children. In children, unlike adults, eye surgery nearly always requires general anesthesia, even for brief procedures. Recommendations for preoperative anxiolysis, fasting guidelines, and management of upper respiratory infections are discussed. Airway considerations and the oculocardiac reflex are highlighted. The prevention of postoperative complications, including those related to opioid prescription therapy, is also addressed. Finally, given the 2016 warning from the Food and Drug Administration about anesthesia neurotoxicity in children, we discuss recent studies on anesthetic neurotoxicity in children undergoing general anesthesia.


EphA4-dependent axon guidance is mediated by the RacGAP alpha2-chimaerin.
Neuronal network formation in the developing nervous system is dependent on the accurate navigation of nerve cell axons and dendrites, which is controlled by attractive and repulsive guidance cues. Ephrins and their cognate Eph receptors mediate many repulsive axonal guidance decisions by intercellular interactions resulting in growth cone collapse and axon retraction of the Eph-presenting neuron. We show that the Rac-specific GTPase-activating protein alpha2-chimaerin binds activated EphA4 and mediates EphA4-triggered axonal growth cone collapse. alpha-Chimaerin mutant mice display a phenotype similar to that of EphA4 mutant mice, including aberrant midline axon guidance and defective spinal cord central pattern generator activity. Our results reveal an alpha-chimaerin-dependent signaling pathway downstream of EphA4, which is essential for axon guidance decisions and neuronal circuit formation in vivo.


Exercise and fall prevention self-management to reduce mobility-related disability and falls after fall-related lower limb fracture in older people: protocol for the RESTORE (Recovery Exercises and STepping On afteR fracturE) randomised controlled trial.
Lasting disability and further falls are common and costly problems in older people following fall-related lower limb and pelvic fractures. Exercise interventions can improve mobility after fracture and reduce falls in older people, however the optimal approach to rehabilitation after fall-related lower limb and pelvic fracture is unclear. This randomised controlled trial aims to evaluate the effects of an exercise and fall prevention self-management intervention on mobility-related disability and falls in older people following fall-related lower limb or pelvic fracture. Cost-effectiveness of the intervention will also be investigated. A randomised controlled trial with concealed allocation, assessor blinding for physical performance tests and intention-to-treat analysis will be conducted. Three hundred and fifty people aged 60 years and over with a fall-related lower limb or pelvic fracture, who are living at home or in a low care residential aged care facility and have completed active rehabilitation, will be recruited. Participants will be randomised to receive a 12-month intervention or usual care. The intervention group will receive ten home visits from a physiotherapist to prescribe an individualised exercise program with motivational interviewing, plus fall prevention education through individualised advice from the physiotherapist or attendance at the group based "Stepping On" program (seven two-hour group sessions). Participants will be followed for a 12-month period. Primary outcome measures will be mobility-related disability and falls. Secondary outcomes will include measures of balance and mobility, falls risk, physical activity, walking aid use, frailty, pain, nutrition, falls efficacy, mood, positive and negative affect, quality of life, assistance required, hospital readmission, and health-system and community-service contact. This study will determine the effect and cost-effectiveness of this exercise self management intervention on mobility-related disability and falls in older people who have recently sustained a fall-related lower limb or pelvic fracture. The results will have implications for the design and implementation of interventions for older people with fall related lower limb fractures. The findings of this study will be disseminated in peer-reviewed journals and through professional and scientific conferences. Australian New Zealand Clinical Trials Registry: ACTRN12610000805077.


Neurologic outcomes after coronary artery bypass grafting with and without cardiopulmonary bypass.
Neurologic injury, in the form of either stroke or more subtle neurocognitive impairment, is a frequent and potentially devastating complication of coronary artery bypass grafting (CABG). The etiology of CABG-associated neurologic injury is likely multifactorial, with the phenomena of cerebral hypoperfusion and embolism being the major contributors. Several perioperative strategies have been developed in an effort to reduce the incidence of CABG-associated neurologic complications. Hypothermic cerebral perfusion, alpha stat acid-base management, and slow patient rewarming have been shown by several investigators to minimize adverse neurologic sequelae associated with the use of cardiopulmonary bypass. Performing CABG without cardiopulmonary bypass (off-pump CABG), meanwhile, has been shown to reduce the risk of perioperative stroke, especially in high-risk patients such as the elderly. Whether off-pump CABG reduces the incidence of less severe neurocognitive impairment has not yet been clearly established and merits further investigation in the form of large, multicenter, randomized trials. Other technical innovations, such as the use of sutureless and clampless aortic anastomotic devices, also may be able to further minimize the neurologic complications associated with CABG.


Clinical features of cats diagnosed with coccidioidomycosis in Arizona, 2004-2018.
The goal of this study was to describe the clinical presentation, diagnosis and treatment of coccidioidomycosis in cats residing in a region endemic for Coccidioides species. A retrospective review of records was performed at both primary and tertiary care veterinary practices in Tucson and Phoenix, Arizona, USA. Data collected included signalment, clinical signs, physical examination findings, diagnostic test results, treatment and outcome. Fifty-one feline cases were identified from six veterinary hospitals. Cats presented with clinical signs and laboratory abnormalities similar to what has been seen in dogs, including respiratory illness (n = 20/51), neutrophilia (n = 24/31), monocytosis (n = 17/31) and hyperglobulinemia (n = 16/30). However, cats at diagnosis were typically significantly ill, with 31/51 having disseminated infection, most commonly to the skin (n = 22). Additionally, 43/44 cats that had serum antibody tests performed were positive, and median titer at diagnosis was 1:32 (range 1:4 to ⩾1:256). Serum antibody titers were significantly reduced (P ⩽0.001) in cats that responded to treatment compared with cats that did not clinically improve. Forty of 46 cats that were treated with oral fluconazole responded and did not require additional therapy. Fourteen cats developed recurrent disease and all but one had antifungal therapy successfully reinstituted. Coccidioidomycosis is a disease of concern for cats residing in the regions endemic for Coccidioides species. Disease is most often disseminated at the time of diagnosis, possibly due to delays in presentation for care and recognition of the infection. Suspicion of disease, serum chemistries, blood cell counts, presence of antibody and imaging aid in the diagnosis of coccidioidomycosis in cats. Serum antibody reduction during treatment frequently correlated with an adequate response to medication. Consideration of coccidioidomycosis as a cause of illness will lead to earlier diagnosis and potentially better treatment outcomes in cats.


A comparison of sedimentary DNA and pollen from lake sediments in recording vegetation composition at the Siberian treeline.
Reliable information on past and present vegetation is important to project future changes, especially for rapidly transitioning areas such as the boreal treeline. To study past vegetation, pollen analysis is common, while current vegetation is usually assessed by field surveys. Application of detailed sedimentary DNA (sedDNA) records has the potential to enhance our understanding of vegetation changes, but studies systematically investigating the power of this proxy are rare to date. This study compares sedDNA metabarcoding and pollen records from surface sediments of 31 lakes along a north-south gradient of increasing forest cover in northern Siberia (Taymyr peninsula) with data from field surveys in the surroundings of the lakes. sedDNA metabarcoding recorded 114 plant taxa, about half of them to species level, while pollen analyses identified 43 taxa, both exceeding the 31 taxa found by vegetation field surveys. Increasing Larix percentages from north to south were consistently recorded by all three methods and principal component analyses based on percentage data of vegetation surveys and DNA sequences separated tundra from forested sites. Comparisons of the ordinations using procrustes and protest analyses show a significant fit among all compared pairs of records. Despite similarities of sedDNA and pollen records, certain idiosyncrasies, such as high percentages of Alnus and Betula in all pollen and high percentages of Salix in all sedDNA spectra, are observable. Our results from the tundra to single-tree tundra transition zone show that sedDNA analyses perform better than pollen in recording site-specific richness (i.e., presence/absence of taxa in the vicinity of the lake) and perform as well as pollen in tracing vegetation composition.


Synaptopodin Is Dispensable for Normal Podocyte Homeostasis but Is Protective in the Context of Acute Podocyte Injury.
Synaptopodin (Synpo) is an actin-associated protein in podocytes and dendritic spines. Many functions in regulating the actin cytoskeleton via RhoA and other pathways have been ascribed to Synpo, yet no pathogenic mutations in the SYNPO gene have been discovered in patients. Naturally occurring Synpo isoforms are known (Synpo-short and -long), and a novel truncated version (Synpo-T) is upregulated in podocytes from Synpo mutant mice. Synpo-T maintains some Synpo functions, which may prevent a podocyte phenotype from emerging in unchallenged mutant mice. Novel mouse models were generated to further investigate the functions of Synpo. In one, CRISPR/Cas9 deleted most of the Synpo gene, preventing production of any detectable Synpo protein. Two other mutant strains made truncated versions of the protein. Adriamycin injections were used to challenge the mice, and Synpo functions were investigated in primary cultured podocytes. Mice that could not make detectable Synpo (Synpo-/- ) did not develop any kidney abnormalities up to 12 months of age. However, Synpo-/- mice were more susceptible to Adriamycin nephropathy. In cultured primary podocytes from mutant mice, the absence of Synpo caused loss of stress fibers, increased the number and size of focal adhesions, and impaired cell migration. Furthermore, loss of Synpo led to decreased RhoA activity and increased Rac1 activation. In contrast to previous findings, podocytes can function normally in vivo in the absence of any Synpo isoform. Synpo plays a protective role in the context of podocyte injury through its involvement in actin reorganization and focal adhesion dynamics.


A survey on the attitudes towards research in medical school.
An observed decrease of physician scientists in medical practice has generated much recent interest in increasing the exposure of research programs in medical school. The aim of this study was to review the experience and attitudes regarding research by medical students in Canada. An anonymous, cross-sectional, self-report questionnaire was administered to second and fourth year students in three medical schools in Ontario between February and May of 2005. Questions were primarily closed-ended and consisted of Likert scales. Descriptive and correlative statistics were used to analyze the responses between students of different years and previous research experience. There was a 47% (327/699) overall response rate to the questionnaire. Despite 87% of respondents reporting that they had been involved in some degree of research prior to medical school, 43% report that they have not been significantly involved in research activity during medical school and 24% had no interest in any participation. There were significant differences in the attitudes towards research endeavors during medical school between students in their fourth year compared to second year. The greatest barriers to involvement in research in medical school appear to be time, availability of research mentors, formal teaching of research methodology and the perception that the student would not receive appropriate acknowledgement for work put towards a research project. The results of this self-report survey outline the significant differences in attitudes towards mandatory research as a component of critical inquiry and scholarship in the undergraduate curriculum in Ontario medical schools.


Effects of ankle extensor muscle afferent inputs on hip abductor and adductor activity in the decerebrate walking cat.
Electrical stimulation of the lateral gastrocnemius-soleus (LGS) nerve at group I afferent strength leads to adaptations in the amplitude and timing of extensor muscle activity during walking in the decerebrate cat. Such afferent feedback in the stance leg might result from a delay in stance onset of the opposite leg. Concomitant adaptations in hip abductor and adductor activity would then be expected to maintain lateral stability and balance until the opposite leg is able to support the body. As many hip abductors and adductors are also hip extensors, we hypothesized that stimulation of the LGS nerve at group I afferent strength would produce increased activation and prolonged burst duration in hip abductor and adductor muscles in the premammillary decerebrate walking cat. LGS nerve stimulation during the extensor phase of the locomotor cycle consistently increased burst amplitude of the gluteus medius and adductor femoris muscles, but not pectineus or gracilis. In addition, LGS stimulation prolonged the burst duration of both gluteus medius and adductor femoris. Unexpectedly, long-duration LGS stimulus trains resulted in two distinct outcomes on the hip abductor and adductor bursting pattern: 1) a change of burst duration and timing similar to medial gastrocnemius; or 2) to continue rhythmically bursting uninterrupted. These results indicate that activation of muscle afferents from ankle extensors contributes to the regulation of activity of some hip abductor and adductor muscles, but not all. These results have implications for understanding the neural control of stability during locomotion, as well as the organization of spinal locomotor networks.


Haemoglobinopathies in tribal populations of India.
Haemoglobinopathies particularly haemoglobin S and E (HbS, HbE) and β-thalassaemia are important challenges for tribal populations in India. The HbS, HbE and β-thalassaemia genes are variably distributed across various tribal populations of India. HbE is mainly restricted in tribals of North-East, West Bengal, Odisha and those in Andaman and Nicobar islands. HbS has more extensive distribution in the country (10-40% trait frequency) and the homozygotes and double heterozygotes present with a wide array of morbidities. The morbidity varies greatly in different areas of the country due to differential co-inheritance of α-thalassaemia gene and interaction of various epistatic and environmental factors. Though substantial data on prevalence of these disorders exist, there is an urgent need to develop integrated hierarchical core facilities to manage the disease. Such centres will generate more data and will also explore areas of management which need more local attention. Newborn screening, genetic counselling, carrier detection, prenatal diagnosis along with management of cases should form the basic infrastructure of haemoglobinopathy management. Research in this areas should continue focusing on various challenges in care delivery, prevention and basic sciences on interaction of haemoglobinopathies with various other infections.


Defect Dominated Charge Transport and Fermi Level Pinning in MoS2/Metal Contacts.
Understanding the electronic contact between molybdenum disulfide (MoS2) and metal electrodes is vital for the realization of future MoS2-based electronic devices. Natural MoS2 has the drawback of a high density of both metal and sulfur defects and impurities. We present evidence that subsurface metal-like defects with a density of ∼1011 cm-2 induce negative ionization of the outermost S atom complex. We investigate with high-spatial-resolution surface characterization techniques the effect of these defects on the local conductance of MoS2. Using metal nanocontacts (contact area < 6 nm2), we find that subsurface metal-like defects (and not S-vacancies) drastically decrease the metal/MoS2 Schottky barrier height as compared to that in the pristine regions. The magnitude of this decrease depends on the contact metal. The decrease of the Schottky barrier height is attributed to strong Fermi level pinning at the defects. Indeed, this is demonstrated in the measured pinning factor, which is equal to ∼0.1 at defect locations and ∼0.3 at pristine regions. Our findings are in good agreement with the theoretically predicted values. These defects provide low-resistance conduction paths in MoS2-based nanodevices and will play a prominent role as the device junction contact area decreases in size.


Oral tolerance inhibits pulmonary eosinophilia in a cockroach allergen induced model of asthma: a randomized laboratory study.
Antigen desensitization through oral tolerance is becoming an increasingly attractive treatment option for allergic diseases. However, the mechanism(s) by which tolerization is achieved remain poorly defined. In this study we endeavored to induce oral tolerance to cockroach allergen (CRA: a complex mixture of insect components) in order to ameliorate asthma-like, allergic pulmonary inflammation. We compared the pulmonary inflammation of mice which had received four CRA feedings prior to intratracheal allergen sensitization and challenge to mice fed PBS on the same time course. Respiratory parameters were assessed by whole body unrestrained plethysmography and mechanical ventilation with forced oscillation. Bronchoalveolar lavage fluid (BAL) and lung homogenate (LH) were assessed for cytokines and chemokines by ELISA. BAL inflammatory cells were also collected and examined by light microscopy. CRA feeding prior to allergen sensitization and challenge led to a significant improvement in respiratory health. Airways hyperreactivity measured indirectly via enhanced pause (Penh) was meaningfully reduced in the CRA-fed mice compared to the PBS fed mice (2.3 ± 0.4 vs 3.9 ± 0.6; p = 0.03). Directly measured airways resistance confirmed this trend when comparing the CRA-fed to the PBS-fed animals (2.97 ± 0.98 vs 4.95 ± 1.41). This effect was not due to reduced traditional inflammatory cell chemotactic factors, Th2 or other cytokines and chemokines. The mechanism of improved respiratory health in the tolerized mice was due to significantly reduced eosinophil numbers in the bronchoalveolar lavage fluid (43300 ± 11445 vs 158786 ± 38908; p = 0.007) and eosinophil specific peroxidase activity in the lung homogenate (0.59 ± 0.13 vs 1.19 ± 0.19; p = 0.017). The decreased eosinophilia was likely the result of increased IL-10 in the lung homogenate of the tolerized mice (6320 ± 354 ng/mL vs 5190 ± 404 ng/mL, p = 0.02). Our results show that oral tolerization to CRA can improve the respiratory health of experimental mice in a CRA-induced model of asthma-like pulmonary inflammation by reducing pulmonary eosinophilia.


The association of immediate post cardiac arrest diastolic hypertension and survival following pediatric cardiac arrest.
In-hospital cardiac arrest occurs in >5000 children each year in the US and almost half will not survive to discharge. Animal data demonstrate that an immediate post-resuscitation burst of hypertension is associated with improved survival. We aimed to determine if systolic and diastolic invasive arterial blood pressures immediately (0-20 min) after return of spontaneous circulation (ROSC) are associated with survival and neurologic outcomes at hospital discharge. This is a secondary analysis of the Pediatric Intensive Care Quality of CPR (PICqCPR) study of invasively measured blood pressures during intensive care unit CPR. Patients were eligible if they achieved ROSC and had at least one invasively measured blood pressure within the first 20 min following ROSC. Post-ROSC blood pressures were normalized for age, sex and height. "Immediate hypertension" was defined as at least one systolic or diastolic blood pressure >90th percentile. The primary outcome was survival to hospital discharge. Of 102 children, 70 (68.6%) had at least one episode of immediate post-CPR diastolic hypertension. After controlling for pre-existing hypotension, duration of CPR, calcium administration, and first documented rhythm, patients with immediate post-CPR diastolic hypertension were more likely to survive to hospital discharge (79.3% vs. 54.5%; adjusted OR = 2.93; 95%CI, 1.16-7.69). In this post hoc secondary analysis of the PICqCPR study, 68.6% of subjects had diastolic hypertension within 20 min of ROSC. Immediate post-ROSC hypertension was associated with increased odds of survival to discharge, even after adjusting for covariates of interest.


p53-dependent regulation of mitochondrial energy production by the RelA subunit of NF-κB.
Aberrant activity of the nuclear factor kappaB (NF-κB) transcription factor family, which regulates cellular responses to stress and infection, is associated with many human cancers. In this study, we define a function of NF-κB in regulation of cellular respiration that is dependent upon the tumor suppressor p53. Translocation of the NF-κB family member RelA to mitochondria was inhibited by p53 by blocking an essential interaction with the HSP Mortalin. However, in the absence of p53, RelA was transported into the mitochondria and recruited to the mitochondrial genome where it repressed mitochondrial gene expression, oxygen consumption, and cellular ATP levels. We found mitochondrial RelA function to be dependent on its conserved C-terminal transactivation domain and independent of its sequence-specific DNA-binding ability, suggesting that its function in this setting was mediated by direct interaction with mitochondrial transcription factors. Taken together, our findings uncover a new mechanism through which RelA can regulate mitochondrial function, with important implications for how NF-κB activity and loss of p53 can contribute to changes in tumor cell metabolism and energy production.


Survey of pediatric ophthalmic diagnoses in a teaching hospital in Nigeria.
A review of pediatric cases seen over one year in the eye clinic of the University of Port Harcourt teaching hospital, Rivers state is presented. To ascertain the different diagnoses made in children who attend the eye clinic and the most common with a view to establish guidelines for equipping a functional pediatric ophthalmic clinic to address the problems if any. Records of all patients aged 0-18 years who attended the clinic in the year 2004 were extracted from the outpatient register. The information was analyzed manually with a simple scientific calculator. Atotal number of 846 pediatric patients were seen in the one year period under review. Newly diagnosed patients were 671 in number. Male female ratio was 1:1.07. The largest number of patients seen was between the ages of 13-18 years (n = 320). Refractive error was the most common condition seen with 248 diagnosed with the condition and a percentage of(28.95%) of the total number of patients seen. Vernal conjunctivitis followed with a percentage of 17.96% (n=152). Sixty-two (7.33%) were diagnosed with iritis while 52 were diagnosed to have cataract (6.15%). Thirty-five had glaucoma as the primary diagnosis (4.14%) while 14 children presented with conjunctivitis (1.65%). Other patients had pterygium (n=9, 1.06%), chalazion (n=4, 0.47%) and corneal opacity (n=6, 0.71%). The patients with refractive error and vernal conjunctivitis were mainly girls 56.5% and 25% respectively. Glaucoma mainly affected boys (57.1%) Cataracts were mainly seen in girls (53.8%). The commonest causes of childhood ocular morbidity in this study were refractive error and allergic conjunctivitis accounting for almost 50% of pediatric patients presenting at the ophthalmology clinic of UPTH. These require prompt pediatric ophthalmic consultation for effective treatment and prevention of avoidable sequelae. Therefore facilities need to be made available to adequately detect these conditions early particularly for refractive errors before amblyopia develops through training of teachers in all schools and public awareness campaigns. It may be important to ensure that legislation be passed to ensure compulsory eye checks before parents enroll their wards in school.


Induction of regulatory T cells by infliximab in Behcet's disease.
To determine whether infliximab induces the development of Foxp3+ T regulatory (Treg) cells in patients with Behçet's disease. The subjects were patients with refractory uveitis caused by Behçet's disease, Vogt-Koyanagi-Harada disease, or ocular toxoplasmosis and healthy volunteers. Purified CD4+ T cells were obtained from patients with uveitis who were treated with colchicine, cyclosporine, or infliximab. Flow cytometry was used to analyze the expression of Foxp3 on CD4+ T cells. Foxp3 was expressed in a small percentage of CD4+ T cells (<5%) from healthy subjects and from patients with active uveitis caused by Behçet's disease, Vogt-Koyanagi-Harada disease, or ocular toxoplasmosis. The percentage of Foxp3+ cells among CD4+ T cells was significantly decreased in patients with active uveitis compared with patients with inactive uveitis in the remission stage. Foxp3 was expressed at a similar level in CD4+ T cells from healthy donors, colchicine-treated patients, and cyclosporine-treated patients, whereas infliximab-treated patients expressed much higher percentages of Foxp3+ cells. Patients who had a high population of Foxp3+ cells during infliximab treatment did not experience any subsequent episodes of acute uveitis. On the other hand, patients with a low population of Foxp3+ cells did experience ocular inflammatory episodes. T cells exposed to infliximab in vitro greatly expressed Foxp3 and produced TGFβ, and the Treg cells significantly suppressed the activation of bystander T cells in vitro. Anti-TNF-α therapy may be useful for patients with ocular complications of Behçet's disease who have a decreased percentage of peripheral Treg cells.


Bacterial and fungal flora of dust deposits in a pig building.
The purpose of the study was to investigate the bacterial and fungal flora of dust deposits in a newly built pig grower finisher building. Viable bacterial counts and microbial species found in a barn which had never housed pigs were compared with those in a barn housing 144 pigs. The quantitative streak plate method was used to measure viable bacterial counts on nutrient agar or sheep blood agar. Viable bacterial counts of the dust deposits were expressed as the number of colony forming units (CFUs)/mg of dust. Gram positive cocci and Gram negative bacilli were identified by an automated system. Identifications with a confidence interval > 90% were accepted at the species level. Fungi were identified to the genus level with slide culture preparations on cereal agar. The lowest viable bacterial count (4.8 x 10(4)/mg of dust) was found in the barn with no pigs. In the barn with pigs the highest viable bacterial count (2.1 x 10(6)/mg of dust) was in dust from the top of a partition close to pig activity. Six species of bacteria or fungi were found in dust from the room with no pigs, whereas 22 different microorganisms were detected in dust from the room with pigs. With the exception of Enterobacter agglomerans no other species of the family Enterobacteriaceae was found in dust deposits in this new pig building. Twelve species of Gram positive bacteria were found in the room housing pigs. The pig is not only a source but also a disperser of airborne bacteria in pig buildings. Speciation of the microbial flora in dust from the pig building suggests that many of the microorganisms were either of human or environmental origin. Nevertheless as some of these microorganisms are known opportunistic pathogens or allergens and because of the documented increased incidence of chronic respiratory symptoms in pig workers, precautions to reduce inhalation of microbial or dust particles by pig workers seem prudent.


Cooperation in an evolutionary prisoner's dilemma on networks with degree-degree correlations.
We study the effects of degree-degree correlations on the success of cooperation in an evolutionary prisoner's dilemma played on a random network. When degree-degree correlations are not present, the standardized variance of the network's degree distribution has been shown to be an accurate analytical measure of network heterogeneity that can be used to predict the success of cooperation. In this paper, we use a local-mechanism interpretation of standardized variance to give a generalization to graphs with degree-degree correlations. Two distinct mechanisms are shown to influence cooperation levels on these types of networks. The first is an intrinsic measurement of base-line heterogeneity coming from the network's degree distribution. The second is the increase in heterogeneity coming from the degree-degree correlations present in the network. A strong linear relationship is found between these two parameters and the average cooperation level in an evolutionary prisoner's dilemma on a network.


Purification of the gene 43, 44, 45, and 62 proteins of the bacteriophage T4 DNA replication apparatus.
A procedure has been developed which allows the T4 bacteriophage proteins corresponding to the products of genes 43, 44, 45, and 62 to be purified to near homogeneity from a single T4-infected cell lysate (greater than 90% single species as judged by sodium dodecyl sulfate polyacrylamide elctrophoresis). In these preparations, the major problem of removing all contaminating nucleases has been overcome. Each of the above proteins is known from genetic analysis to be essential for phage DNA replication. The protein product of gene 43 is T4 DNA polymerase, and its recovery can be monitored using a standard DNA polymerase assay. The other three gene products have been designated as "polymerase accessory proteins," since they directly enhance polymerase function on both single- and double-stranded DNA templates. Their activities were monitored by an "in vitro complementation assay," which measures the stimulation of DNA synthesis observed in a concentrated lysate of T4 mutant-infected Escherichia coli cells when the missing T4 wild type protein is added. Starting from 300 g of infected cell paste, we obtained 9.3 mg of gene 43 protein, 21 mg of gene 45 protein, and 70 mg of a tight complex made up of 44 and 62 proteins; final yields were estimated at 30%, 14%, and 28%, respectively, of the initial activity present in the lysate. When the above purified proteins are incubated with preparations of two other T4 DNA replication proteins (gene 41 and gene 32 proteins) plus deoxyribonucleoside and ribonucleoside triphosphates, extensive DNA synthesis occurs on both single- and double-stranded DNA templates. As reported elsewhere, this synthesis mimicks that catalyzed by the T4 DNA replication apparatus in vivo.


period and timeless mRNA Splicing Profiles under Natural Conditions in Drosophila melanogaster.
Previous analysis of Drosophila circadian behavior under natural conditions has revealed a number of novel and unexpected features. Here we focus on the oscillations of per and tim mRNAs and their posttranscriptional regulation and observe significant differences in molecular cycling under laboratory and natural conditions. In particular, robust per mRNA cycling from fly heads is limited to the summers, whereas tim RNA cycling is observed throughout the year. When both transcripts do cycle, their phases are similar, except for the very warmest summer months. We also study the natural splicing profiles of per and tim transcripts and observe a clear relationship between temperature and splicing. In natural conditions, we confirm the relationship between accumulation of the per(spliced) variant, low temperature, and the onset of the evening component of locomotor activity, first described in laboratory conditions. Intriguingly, in the case of tim splicing, we detect the opposite relationship, with tim(spliced) expression increasing at higher temperatures. A first characterization of the 4 different TIM protein isoforms (resulting from the combination of the natural N-terminus length polymorphism and the C-terminus alternative splicing) using the 2-hybrid assay showed that the TIM(unspliced) isoforms have a stronger affinity for CRY, but not for PER, suggesting that the tim 3' splicing could have physiological significance, possibly in temperature entrainment and/or adaptation to seasonal environments.


Phosphorylation of Ser136 is critical for potent bone sialoprotein-mediated nucleation of hydroxyapatite crystals.
Acidic phosphoproteins of mineralized tissues such as bone and dentin are believed to play important roles in HA (hydroxyapatite) nucleation and growth. BSP (bone sialoprotein) is the most potent known nucleator of HA, an activity that is thought to be dependent on phosphorylation of the protein. The present study identifies the role phosphate groups play in mineral formation. Recombinant BSP and peptides corresponding to residues 1-100 and 133-205 of the rat sequence were phosphorylated with CK2 (protein kinase CK2). Phosphorylation increased the nucleating activity of BSP and BSP-(133-205), but not BSP-(1-100). MS analysis revealed that the major site phosphorylated within BSP-(133-205) was Ser136, a site adjacent to the series of contiguous glutamate residues previously implicated in HA nucleation. The critical role of phosphorylated Ser136 in HA nucleation was confirmed by site-directed mutagenesis and functional analyses. Furthermore, peptides corresponding to the 133-148 sequence of rat BSP were synthesized with or without a phosphate group on Ser136. As expected, the phosphopeptide was a more potent nucleator. The mechanism of nucleation was investigated using molecular-dynamics simulations analysing BSP-(133-148) interacting with the {100} crystal face of HA. Both phosphorylated and non-phosphorylated sequences adsorbed to HA in extended conformations with alternating residues in contact with and facing away from the crystal face. However, this alternating-residue pattern was more pronounced when Ser136 was phosphorylated. These studies demonstrate a critical role for Ser136 phosphorylation in BSP-mediated HA nucleation and identify a unique mode of interaction between the nucleating site of the protein and the {100} face of HA.


Inner and Outer Retinal Contributions to Pupillary Light Response: Correlation to Functional and Morphologic Parameters in Glaucoma.
To evaluate in patients with primary open-angle glaucoma (POAG) the contribution of the inner and outer retinal photoreceptors to the pupillary light responses (PLRs) correlated with both functional (color vision and visual field perimetry) and morphologic (optical coherence tomography) parameters. In total, 45 patients with POAG and 25 healthy control participants were evaluated. The PLR was measured as pupil diameter with an eye tracker; stimuli were presented in a Ganzfeld. Pupil responses were measured monocularly, to 1 second blue (470 nm) and red (640 nm) flashes with -3, -2, -1, 0, 1, 2, and 2.4 log cd/m luminance levels. Color vision was evaluated with the Cambridge Color Test, visual field was measured by standard automatic perimetry, and retinal nerve fiber layer thickness was evaluated by optical coherence tomography. Patients with moderate and severe POAG have a significantly decreased PLR that depends on the severity of POAG, for both the 470 and 640 nm stimuli, revealing the reduction of the contributions of the rods, cones, and intrinsically photosensitive retinal ganglion cells to PLR. A significant loss of color discrimination along the blue-yellow axis was observed in all stages of POAG. Correlations among standard automatic perimetry, retinal nerve fiber layer thickness, Cambridge Color Test, PLR, and melanopsin parameters were found. The results provide evidence that in moderate and severe stages of POAG, both the inner and outer retinal contributions to PLR are affected. Also, a worsening in color vision was correlated with reduced PLR responses at high-intensity stimuli. These findings may enhance the clinical management of POAG patients.


Cervical Necrotizing Fasciitis and Shock in the Post-Operative Pediatric Patient.
Cervical necrotizing fasciitis (CNF) is a rare infection that can quickly lead to devastating patient outcomes. Considering the vital importance of surrounding neck structures, rapid control of the infection is essential. Infection is most frequently polymicrobial and occurs in the adult patient in the context of certain medical comorbidities. Complications are typically limited to tissue destruction in the form of necrosis. There are no current reports describing CNF complicated by acute shock in the post-operative pediatric patient. Here, we present a pediatric case of CNF complicated by acute shock following Sistrunk procedure for removal of a supposed thyroglossal duct cyst. This case illustrates a potential post-operative complication that can be seen within the pediatric patient. Although most reported examples of CNF are polymicrobial and result from odontogenic infection, providers should be aware of other potential sources of disease. It is important for the pediatric surgeon to rapidly identify CNF and consequent shock, as prompt medical and surgical interventions are critical to offering the best chance of patient survival.


The nuclear pore complex mediates binding of the Mig1 repressor to target promoters.
All eukaryotic cells alter their transcriptional program in response to the sugar glucose. In Saccharomyces cerevisiae, the best-studied downstream effector of this response is the glucose-regulated repressor Mig1. We show here that nuclear pore complexes also contribute to glucose-regulated gene expression. NPCs participate in glucose-responsive repression by physically interacting with Mig1 and mediating its function independently of nucleocytoplasmic transport. Surprisingly, despite its abundant presence in the nucleus of glucose-grown nup120Δ or nup133Δ cells, Mig1 has lost its ability to interact with target promoters. The glucose repression defect in the absence of these nuclear pore components therefore appears to result from the failure of Mig1 to access its consensus recognition sites in genomic DNA. We propose that the NPC contributes to both repression and activation at the level of transcription.


Evaluation of a Serious Self-Regulation Game Intervention for Overweight-Related Behaviors ("Balance It"): A Pilot Study.
Serious games have the potential to promote health behavior. Because overweight is still a major issue among secondary vocational education students in the Netherlands, this study piloted the effects of "Balance It," a serious self-regulation game intervention targeting students' overweight-related behaviors: dietary intake and physical activity (PA). We aimed to pilot the effects of Balance It on secondary vocational education students' dietary intake and PA. In total, 501 secondary vocational education students participated at baseline (intervention: n=250; control: n=251) in this pre-post cluster randomized trial. After 4 weeks, at immediate posttest, 231 students filled in the posttest questionnaire (intervention: n=105; control: n=126). The sample had a mean age of 17.28 (SD 1.26, range 15-21) years, 62.8% (145/231) were female, and 26.8% (62/231) had a non-Dutch background. Body mass index (BMI kg/m2) ranged from 14.4 to 31.1 (mean 21.1, SD 3.3). The intervention and control groups were compared on the primary (behavioral) outcomes of dietary intake (fruit and vegetable consumption, snack consumption, and soft drink consumption) and PA (moderate and vigorous). Additionally, we explored (1) differences between the intervention and control groups in determinants of dietary intake and PA, including attitude, self-efficacy, intention, barrier identification, action planning, and action control, and (2) differences between active (intervention) users and the control group in dietary intake, PA, and associated determinants. After corrections for multiple testing, we did not find significant differences between the intervention group and control group in terms of dietary intake, PA, and determinants of dietary intake and PA. Exploratory research indicated that only 27.6% (29/105) of the intervention group reported actual intervention use (ie, active users). For exploratory reasons, we compared the active users (n=29) with the control group (n=124) and corrected for multiple testing. Results showed that active users' snack consumption decreased more strongly (active users: mean change=-0.20; control group: mean change=-0.08; beta=-0.36, P=.01, R2 change=.05), and their use of active transport had a stronger increase (active users: mean change=0.92; control group=-0.12; beta=1.58, P=.02, R2 change=.03) than the control group. Results also revealed significant differences in action planning (active users: mean change=0.42; control group: mean change=0.07; beta=0.91, P=.01, R2 change=.04) and action control (active users: mean change=0.63; control group: mean change=-0.05; beta=1.25, P=.001, R2 change=.08) in terms of unhealthy eating. The Balance It intervention did not show favorable effects on dietary intake and PA compared to the control condition. However, only a small number of people in the intervention condition actually used Balance It (27.6%). Exploratory analyses did suggest that, if used as planned, Balance It could contribute to changing dietary intake and PA behaviors, albeit it remains debatable whether this would be sufficient to prevent overweight.


OTUD4 Is a Phospho-Activated K63 Deubiquitinase that Regulates MyD88-Dependent Signaling.
Ubiquitination is a major mechanism that regulates numerous cellular processes, including autophagy, DNA damage signaling, and inflammation. While hundreds of ubiquitin ligases exist to conjugate ubiquitin onto substrates, approximately 100 deubiquitinases are encoded by the human genome. Thus, deubiquitinases are likely regulated by unidentified mechanisms to target distinct substrates and cellular functions. Here, we demonstrate that the deubiquitinase OTUD4, which nominally encodes a K48-specific deubiquitinase, is phosphorylated near its catalytic domain, activating a latent K63-specific deubiquitinase. Besides phosphorylation, this latter activity requires an adjacent ubiquitin-interacting motif, which increases the affinity of OTUD4 for K63-linked chains. We reveal the Toll-like receptor (TLR)-associated factor MyD88 as a target of this K63 deubiquitinase activity. Consequently, TLR-mediated activation of NF-κB is negatively regulated by OTUD4, and macrophages from Otud4-/- mice exhibit increased inflammatory signaling upon TLR stimulation. Our results reveal insights into how a deubiquitinase may modulate diverse processes through post-translational modification.


Group II intron from Pseudomonas alcaligenes NCIB 9867 (P25X): entrapment in plasmid RP4 and sequence analysis.
Pseudomonas alcaligenes NCIB 9867 (strain P25X), which grows on 2,5-xylenol and harbours the plasmid RP4, was mated with a plasmid-free derivative of Pseudomonas putida NCIB 9869, strain RA713, which cannot grow on 2,5-xylenol. Some RA713 transconjugants, initially selected on 2,5-xylenol, were found to carry RP4 plasmids that had acquired additional fragments (designated Xin) which ranged in size from 2 kb to approximately 26 kb instability of DNA inserts in RP4::Xin hybrid plasmids was observed. The smallest insert present in a stable RP4::Xin6 hybrid plasmid, termed Xin6, yielded multiple bands when it was used as a probe with digested P25X chromosomal DNA. Sequence analysis of Xin6 led to the discovery of an open reading frame with homology to the maturases of group II introns. The Xin6 insert also exhibited several features characteristic of a group II intron. These included the presence of the consensus sequence GUGYG at the 5' and and RAY at the 3' end of the intron. RNA secondary structure modelling of Xin6 also revealed the presence of perfectly conserved domains V and VI. Differences were detected in the Xin6 hybridization profiles of several P25X catabolic mutants that have lost the ability to grow on 2,5-xylenol. In these mutants the loss of 2,5-xylenol degradative ability could be due to genome rearrangements mediated by sequences related to the Xin6 group II intron. This is the first reported group II intron isolated from Pseudomonas spp. and the first time that the mobility of a bacterial group II intron has been demonstrated.


SIRT1 genetic variation and mortality in type 2 diabetes: interaction with smoking and dietary niacin.
SIRT1 protects cells against oxidative stress and aging. Its activity may be modulated by dietary niacin (vitamin B3) intake. We studied the association of SIRT1 genetic variation with mortality in subjects with increased oxidative stress (type 2 diabetes and smokers) in relation to dietary niacin. In 4573 participants from the Rotterdam Study, including 413 subjects with prevalent and 378 with incident type 2 diabetes, three SIRT1 tagging SNPs were genotyped and all-cause mortality was studied (average follow-up 12 years). We found no association between SIRT1 variation and mortality in the total population or in smokers. In subjects with prevalent type 2 diabetes, homozygous carriers of the most common SIRT1 haplotype, 1, had 1.5 times (95%CI 1.1-2.1) increased mortality risk compared to noncarriers. This risk further increased among smokers and those with low niacin intake. In the lowest tertile of niacin intake, mortality risk was increased 2.3 (95%CI 1.1-4.9) and 5.7 (95%CI 2.5-13.1) times for heterozygous and homozygous carriers of haplotype 1. Subjects with incident diabetes showed similar findings but only when they smoked. We conclude that in subjects with type 2 diabetes, SIRT1 genetic variation influences survival in interaction with dietary niacin and smoking. Correction of niacin deficiency and SIRT1 modulators may prolong the life span of patients with diabetes.


Willingness to use functional breads. Applying the Health Belief Model across four European countries.
The present study focused on the role of the Health Belief Model (HBM) in predicting willingness to use functional breads, across four European countries: UK (N=552), Italy (N=504), Germany (N=525) and Finland (N=513). The behavioural evaluation components of the HBM (the perceived benefits and barriers conceptualized respectively as perceived healthiness and pleasantness) and the health motivation component were good predictors of willingness to use functional breads whereas threat perception components (perceived susceptibility and perceived anticipated severity) failed as predictors. This result was common in all four countries and across products. The role of 'cue to action' was marginal. On the whole the HBM fit was similar across the countries and products in terms of significant predictors (the perceived benefits, barriers and health motivation) with the exception of self-efficacy which was significant only in Finland. Young consumers seemed more interested in the functional bread with a health claim promoting health rather than in reducing risk of disease, whereas the opposite was true for older people. However, functional staple foods, such as bread in this European study, are still perceived as common foods rather than as a means of avoiding diseases. Consumers seek these foods for their healthiness (the perceived benefits) as they expect them to be healthier than regular foods and for the pleasantness (the perceived barriers) as they do not expect any change in the sensory characteristics due to the addition of the functional ingredients. The importance of health motivation in willingness to use products with health claims implies that there is an opening for developing better models for explaining health-promoting food choices that take into account both food and health-related factors without making a reference to disease-related outcome.


Criteria and other methodological considerations in the evaluation of time at V.O2max.
Many previous studies have examined the time limit at which an individual can maintain VO(2max) (T(lim) VO(2max)) during high-intensity continuous and intermittent runs to exhaustion. The main purpose of the present study was to examine the effects of employing different criteria used in previous investigations during T(lim) VO(2max) evaluation. Seven moderately trained competitive runners completed 2 running tests to exhaustion, during which metabolic data was obtained from breath-by-breath gas analysis. The 1(st) test was an incremental test to evaluate VO(2max) and the minimal running velocity at which VO(2max) was elicited (v VO(2max)). The 2(nd) test was a continuous single velocity test at v VO(2max) from which the time to attain VO(2max) (TA VO(2max)) and T(lim) VO(2max) were subsequently evaluated. Time at VO(2max) was evaluated employing 6 specific criteria. Intra-individual differences in T(lim) VO(2max) values due to applying the different criteria were analysed using a one-way ANOVA, with significant differences between pairs identified using Tukey's HSD posthoc test. Significance was set at p<0.05. A one-way ANOVA demonstrated that significant differences (F=4.03, p=0.005) existed between T(lim) VO(2max) values generated by employing the six different criteria. The present study provides support that employing different criteria in the evaluation of T(lim) VO(2max), such as those used in previous investigations, leads to significantly different T(lim) VO(2max) values. However, the practical implications of these measurement differences require further investigation.


Oxygen consumption during pranayamic type of very slow-rate breathing.
To determine whether the yogic Ujjayi pranayamic type of breathing that involves sensory awareness and consciously controlled, extremely slow-rate breathing including at least a period of end-inspiration breath holding in each respiratory cycle would alter oxygen consumption or not, ten males with long standing experience in pranayama, and volunteering to participate in the laboratory study were assessed. These subjects aged 28-59 yr, had normal health appropriate to their age. Since kumbhak (timed breath holding) is considered as an important phase of the respiratory cycle in the pranayama, they were categorised into two groups of five each, one group practising the short kumbhak varieties of pranayama, and the other the long kumbhak varieties of pranayama. The duration of kumbhak phase was on an average 22.2 percent of the respiratory cycle in the short kumbhak group, and 50.4 per cent in the long kumbhak group. The oxygen consumption was measured in test sessions using the closed circuit method of breathing oxygen through the Benedict-Roth spirometer. Each subject was tested in several repeat sessions. Values of oxygen consumption of the period of pranayamic breathing, and of post-pranayamic breathing period, were compared to control value of oxygen consumption of the prepranayamic breathing period of each test session. The results revealed that the short kumbhak pranayamic breathing caused a statistically significant increase (52%) in the oxygen consumption (and metabolic rate) compared to the pre-pranayamic base-line period of breathing. In contrast to the above, the long kumbhak pranayamic breathing caused a statistically significant lowering (19% of the oxygen consumption (and metabolic rate).(ABSTRACT TRUNCATED AT 250 WORDS)


Imaging the distribution of skin lipids and topically applied compounds in human skin using mass spectrometry.
The barrier functions of skin against water loss, microbial invasion and penetration of xenobiotics rely, in part, on the spatial distribution of the biomolecular constituents in the skin structure, particularly its horny layer (stratum corneum). However, all skin layers are important to describe normal and dysfunctional skin conditions, and to develop adapted therapies or skin care products. In this work, time-of-flight secondary ion mass spectrometry (ToF-SIMS) combined with scanning electron microscopy (SEM) was used to image the spatial distribution of a variety of molecular species, from stratum corneum down to dermis, in cross-section samples of human abdominal skin. The results demonstrate the expected localization of ceramide and saturated long-chain fatty acids in stratum corneum (SC) and cholesterol sulfate in the upper part of the viable epidermis. The localization of exogenous compounds is demonstrated by the detection and imaging of carvacrol (a constituent of oregano or thyme essential oil) and ceramide, after topical application onto ex vivo human skin. Carvacrol showed pronounced accumulation to triglyceride-containing structures in the deeper parts of dermis. In contrast, the exogenous ceramide was found to be localized in SC. Furthermore, the complementary character of this approach with classical ex vivo skin absorption analysis methods is demonstrated.


Antiproliferative effect of 1alpha,25-dihydroxyvitamin D3 in human prostate cancer cell line LNCaP involves reduction of cyclin-dependent kinase 2 activity and persistent G1 accumulation.
1Alpha,25-dihydroxyvitamin D3 (1,25 D), the most active metabolite of vitamin D3, exerts antiproliferative and prodifferentiating effects on some human prostate cancer cell lines. We previously reported an inverse relationship between functional vitamin D receptor (VDR) levels and antiproliferative response to 1,25 D in two human prostate cancer cell lines, LNCaP and ALVA 31. Although LNCaP cells are far more sensitive to growth inhibition by 1,25 D than ALVA 31 cells, LNCaP express approximately half the number of VDR as ALVA 31. Two other human prostate cancer cell lines studied, PC3 and DU145, express lower levels of functional VDR and are relatively insensitive to growth inhibition by 1,25 D. In this report, we investigated potential mechanisms of the variable antiproliferative activity of 1,25 D. In PC3 cells stably expressing VDR [PC3(VDR)] at levels comparable to LNCaP, 1,25 D treatment resulted in only moderate growth inhibition. These results further support the contention that VDR expression, although required, is not sufficient for maximal growth suppression by 1,25 D, as is exhibited by LNCaP cells. We did not detect 1,25 D-mediated DNA fragmentation after 4 days of 1,25 D treatment in either LNCaP or ALVA 31 cells. This result suggests that variability in 1,25 D sensitivity does not derive from differences in the capacity of these cells to undergo apoptosis in response to 1,25 D. Flow cytometry of propidium iodine-stained cells revealed that 48 h 1,25 D treatment of LNCaP cells resulted in a 2-fold decrease of cells in G2/M plus S phases and accumulation of LNCaP cells in the G1/G0 phase. This effect persisted for 72 h after 1,25 D removal. In contrast, 1,25 D did not significantly alter the cell cycle distribution of ALVA 31 or PC3(VDR) cells. Consistent with accumulation of cells in G1/G0, 1,25 D treatment of LNCaP cells resulted in decreased retinoblastoma protein phosphorylation, repressed E2F transcriptional activity, increased levels of the cyclin-dependent kinase (CDK) inhibitor p21(WAF1, CIP1), and decreased CDK2 activity. However, p21 messenger RNA levels were not altered, suggesting translational or posttranslational regulation of p21 by 1,25 D. In contrast, p21 was not detected in ALVA 31 or PC3(VDR) and was not induced by 1,25 D, consistent with the failure of 1,25 D to influence cell cycle distribution in these cells. These results suggest that variability in sensitivity to the antiproliferative effects of 1,25 D among prostate cancer cells is dependent, at least in part, on the integrity of the retinoblastoma pathway and in particular on p21 expression and 1,25 D regulation of CDK2 activity.


Delaying mobility disability in people with Parkinson disease using a sensorimotor agility exercise program.
This article introduces a new framework for therapists to develop an exercise program to delay mobility disability in people with Parkinson disease (PD). Mobility, or the ability to efficiently navigate and function in a variety of environments, requires balance, agility, and flexibility, all of which are affected by PD. This article summarizes recent research identifying how constraints on mobility specific to PD, such as rigidity, bradykinesia, freezing, poor sensory integration, inflexible program selection, and impaired cognitive processing, limit mobility in people with PD. Based on these constraints, a conceptual framework for exercises to maintain and improve mobility is presented. An example of a constraint-focused agility exercise program, incorporating movement principles from tai chi, kayaking, boxing, lunges, agility training, and Pilates exercises, is presented. This new constraint-focused agility exercise program is based on a strong scientific framework and includes progressive levels of sensorimotor, resistance, and coordination challenges that can be customized for each patient while maintaining fidelity. Principles for improving mobility presented here can be incorporated into an ongoing or long-term exercise program for people with PD.


The physical functioning and mental health of informal carers: evidence of care-giving impacts from an Australian population-based cohort.
Informal carers represent a substantial proportion of the population in many countries and health is an important factor in their capacity to continue care-giving. This study investigated the impact of care-giving on the mental and physical health of informal carers, taking account of contextual factors, including family and work. We examined health changes from before care-giving commenced to 2 and 4 years after, using longitudinal data from the Household Income and Labour Dynamics in Australia survey. The sample comprised 424 carers and 424 propensity score-matched non-carers. Health was self-assessed, measured with the SF-36 Health Survey Mental Health (MH) and Physical Functioning (PF) scales. Care-giving was classified as non-carer, low (<5 hours/week), moderate (5-19 hours/week) and high (20 or more hours/week). PF and MH change scores were regressed on baseline scores, care-giving, covariates (including work, family and socio-demographic characteristics) and interactions to identify impacts for subgroups. The physical and mental health impacts differed by gender, and care-giving hours and carer work hours were important contextual factors. Deterioration in both PF and MH was worse for females after 2 years and deterioration in MH was worse for males after 4 years. Among carers aged 40-64 years, there was a 17-point decline in PF (P = 0.009) and a 14-point decline in MH (P < 0.0001) after 2 years for female high caregivers working full-time and 9.3 point improvement (P = 0.02) for non-working male high caregivers. Change was not significant for non-carers. The study found that not all carers suffer adverse health impacts; however, the combination of high levels of care-giving with workforce participation can increase the risk of negative physical and mental health effects (particularly in female carers). Working carers providing high levels of care represent a vulnerable subgroup where supportive and preventive services might be focused.


The effect of a garlic preparation on plasma lipid levels in moderately hypercholesterolemic adults.
Lipid management is well established as an effective preventive and management tool for cardiovascular disease (CVD). Health claims regarding the cholesterol lowering benefits of garlic are widespread. However, the clinical trial data are inconsistent. The effect of two doses of a commercial garlic preparation on plasma lipids were evaluated, compared to a placebo, in moderately hypercholesterolemic adults (baseline low density lipoprotein cholesterol (LDL-C)=157. 4+/-18.7, mean+/-S.D.). Fifty-one adults, aged 51.8+/-8.3 years participated in a double-blind, placebo-controlled, parallel treatment trial conducted in an outpatient research clinic. They were randomized to a placebo or a garlic botanical blend providing 500 or 1000 mg dehydrated garlic powder/day (three groups, 16-18 subjects per group). Plasma lipids were assessed every 2 weeks for 12 weeks. The study was designed with sufficient power to detect a 10% relative decline in LDL-C. The absolute mean changes in LDL-C over 12 weeks were 0.0+/-4.3, +1.4+/-4.8, and -10.1+/-6.8 mg/dl for the placebo, half-dose and full-dose, respectively. In the full-dose group, the LDL-C decrease of 6.1% was not significantly different from the other groups (P=0.5). No significant differences were observed for total- or high-density lipoprotein cholesterol (HDL-C), or triacylglycerol levels. In conclusion, the garlic powder preparation used in this study among moderately hypercholesterolemic adults did not significantly effect plasma lipids levels. There was no indication of a graded affect by garlic dose over the range of 0, 500 and 1000 mg/day. A small (<10%) effect on LDL-C levels or a threshold effect requiring larger doses are not eliminated by this study.


Meaningful family relationships: neurocognitive buffers of adolescent risk taking.
Discordant development of brain regions responsible for cognitive control and reward processing may render adolescents susceptible to risk taking. Identifying ways to reduce this neural imbalance during adolescence can have important implications for risk taking and associated health outcomes. Accordingly, we sought to examine how a key family relationship-family obligation-can reduce this vulnerability. Forty-eight adolescents underwent an fMRI scan during which they completed a risk-taking and cognitive control task. Results suggest that adolescents with greater family obligation values show decreased activation in the ventral striatum when receiving monetary rewards and increased dorsolateral PFC activation during behavioral inhibition. Reduced ventral striatum activation correlated with less real-life risk-taking behavior and enhanced dorsolateral PFC activation correlated with better decision-making skills. Thus, family obligation may decrease reward sensitivity and enhance cognitive control, thereby reducing risk-taking behaviors.


Intracellular recording from myenteric neurons of the guinea-pig ileum that respond to stretch.
1. Isolated longitudinal muscle-myenteric plexus preparations from guinea-pig ileum were used to investigate the activity of myenteric neurons when the tissue was stretched in the circumferential direction. Membrane potentials were recorded via flexibly mounted intracellular recording electrodes containing Neurobiotin in 1 M KCl. The preparations were stretched to constant widths (+20% and +40% beyond slack width). 2. Multipolar neurons (Dogiel type II morphology) discharged spontaneous action potentials and proximal process potentials during maintained stretching, three of twenty-one at +20% stretch and seven of nine at +40% stretch. At the maximum extent of stretch tried, +40% beyond slack tissue width, action potentials in Dogiel type II neurons occurred at 10-33 Hz. Neurons with other morphologies were all uniaxonal. Some displayed spontaneous fast EPSPs or action potentials, three of forty one at +20% stretch and seven of nineteen at +40% stretch. 3. In seven of eight Dogiel type II neurons, action potentials or proximal process potentials persisted when membrane hyperpolarization was imposed via the recording electrode. Action potential discharge was abolished by hyperpolarization in seven of nine uniaxonal neurons; the exceptions were two orally projecting neurons. 4. Dogiel type II and uniaxonal neurons were classified as rapidly accommodating if they discharged action potentials only at the beginning of a 500 ms intracellular depolarizing pulse and slowly accommodating if they discharged for more than 250 ms. For Dogiel type II neurons, three of thirteen were slowly accommodating at +20% stretch and two of four at 40% stretch. For uniaxonal neurons the corresponding data were twelve of twenty-six and fifteen of nineteen neurons. The slowly accommodating state was associated with increased cell input resistance in uniaxonal neurons. 5. The spontaneous action potential discharge in Dogiel type II and uniaxonal neurons ceased when the muscle was relaxed pharmacologically by nicardipine (3 microM) or isoprenaline (1 microM), although the applied stretch was maintained. At the same time, evoked spike discharge became rapidly accommodating. 6. We conclude that many Dogiel type II neurons, and possibly some orally projecting uniaxonal neurons, are intrinsic, stretch-sensitive, primary afferent neurons that respond to muscle tension with sustained action potential discharge.


Correlation between ERG Fusion Protein and Androgen Receptor Expression by Immunohistochemistry in Prostate, Possible Role in Diagnosis and Therapy.
Background: Recent discovery of gene rearrangements have brought a new look to the molecular pathogenesis of cancer. Gene fusions occur in nearly 60% of prostate adenocarcinoma, being the TMPRSS2-ERG one of the most common. Evidence supports the role of ERG fusion in tumorigenesis, progression and invasion via effecting pathways such as WNT, MYC, uPA, PI3K/AKT/PTEN, RAS/RAF/MAPF, NKX3.1, GST-pi and androgen receptor (AR) mediated signaling. Most of the ERG fusions involve 5'-partners androgen responsive. Therefore, we aimed to evaluate AR and ERG fusion protein expression on prostate tissue to find clinicopathological applications and possible role in therapy. Methods: One hundred three samples, including prostate core biopsies and radical prostatectomy specimens, were evaluated for ERG and AR expression by immunohistochemistry (IHC). ERG rearrangement was done by fluorescence in situ hybridization (FISH) on 11 randomly selected cases and correlated with IHC results. Results: From the total of 103 samples, eight (8/103) were benign, fourteen (14/103) had atypical glands, two (2/103) had prostatic intraepithelial neoplasia (PIN), and seventy nine (79/103) showed prostate adenocarcinoma. Forty four (44/79) tumor cases were Gleason score (GS) 6-7 (lower GS), and thirty five (35/79) were GS of 8-10 (higher GS). ERG immunoreaction was observed in 27.8% (22/79) of the tumor cases, showing higher expression in those with lower GS (68.2%, 15/22) compared to higher GS (31.8%, 7/22). Neither benign glands nor PIN stained with ERG. AR expression was observed in 75% of benign samples, 78.5% of atypical glands, 100% of PIN, and in 87.3% of tumor cases with no significant difference based on GS. Co-expression of ERG and AR was evaluated on all the tumor samples. ERG+/AR+ was seen in 77.3% (17/22) of the ERG+ tumor cases, with higher frequency in lower GS (64.7%, 11/17) compared to those with higher GS (35.3%, 6/17). All but five corresponding ERG+ tumor samples were negative for AR. Only 5 samples were ERG-/AR- corresponding to adenocarcinoma GS of 6. Presence or absence of ERG rearrangement was confirmed by FISH and correlated with IHC results. Conclusions: Characterization of ERG status by IHC in prostate tissue has an excellent correlation with FISH. It may also assist in diagnosis since none of the benign glands stained with ERG. Co-expression of ERG+/AR+ in prostate tumor by IHC may suggest gene fusion between ERG and a 5'-partner driven by androgen signaling such as TMPRSS2, which it could represent an important ancillary test for clinical management and development of new therapeutic targets.


TIDieR-Placebo: A guide and checklist for reporting placebo and sham controls.
Placebo or sham controls are the standard against which the benefits and harms of many active interventions are measured. Whilst the components and the method of their delivery have been shown to affect study outcomes, placebo and sham controls are rarely reported and often not matched to those of the active comparator. This can influence how beneficial or harmful the active intervention appears to be. Without adequate descriptions of placebo or sham controls, it is difficult to interpret results about the benefits and harms of active interventions within placebo-controlled trials. To overcome this problem, we developed a checklist and guide for reporting placebo or sham interventions. We developed an initial list of items for the checklist by surveying experts in placebo research (n = 14). Because of the diverse contexts in which placebo or sham treatments are used in clinical research, we consulted experts in trials of drugs, surgery, physiotherapy, acupuncture, and psychological interventions. We then used a multistage online Delphi process with 53 participants to determine which items were deemed to be essential. We next convened a group of experts and stakeholders (n = 16). Our main output was a modification of the existing Template for Intervention Description and Replication (TIDieR) checklist; this allows the key features of both active interventions and placebo or sham controls to be concisely summarised by researchers. The main differences between TIDieR-Placebo and the original TIDieR are the explicit requirement to describe the setting (i.e., features of the physical environment that go beyond geographic location), the need to report whether blinding was successful (when this was measured), and the need to present the description of placebo components alongside those of the active comparator. We encourage TIDieR-Placebo to be used alongside TIDieR to assist the reporting of placebo or sham components and the trials in which they are used.


Smooth Muscle-Like Cells Generated from Human Mesenchymal Stromal Cells Display Marker Gene Expression and Electrophysiological Competence Comparable to Bladder Smooth Muscle Cells.
The use of mesenchymal stromal cells (MSCs) differentiated toward a smooth muscle cell (SMC) phenotype may provide an alternative for investigators interested in regenerating urinary tract organs such as the bladder where autologous smooth muscle cells cannot be used or are unavailable. In this study we measured the effects of good manufacturing practice (GMP)-compliant expansion followed by myogenic differentiation of human MSCs on the expression of a range of contractile (from early to late) myogenic markers in relation to the electrophysiological parameters to assess the functional role of the differentiated MSCs and found that differentiation of MSCs associated with electrophysiological competence comparable to bladder SMCs. Within 1-2 weeks of myogenic differentiation, differentiating MSCs significantly expressed alpha smooth muscle actin (αSMA; ACTA2), transgelin (TAGLN), calponin (CNN1), and smooth muscle myosin heavy chain (SM-MHC; MYH11) according to qRT-PCR and/or immunofluorescence and Western blot. Voltage-gated Na+ current levels also increased within the same time period following myogenic differentiation. In contrast to undifferentiated MSCs, differentiated MSCs and bladder SMCs exhibited elevated cytosolic Ca2+ transients in response to K+-induced depolarization and contracted in response to K+ indicating functional maturation of differentiated MSCs. Depolarization was suppressed by Cd2+, an inhibitor of voltage-gated Ca2+-channels. The expression of Na+-channels was pharmacologically identified as the Nav1.4 subtype, while the K+ and Ca2+ ion channels were identified by gene expression of KCNMA1, CACNA1C and CACNA1H which encode for the large conductance Ca2+-activated K+ channel BKCa channels, Cav1.2 L-type Ca2+ channels and Cav3.2 T-type Ca2+ channels, respectively. This protocol may be used to differentiate adult MSCs into smooth muscle-like cells with an intermediate-to-late SMC contractile phenotype exhibiting voltage-gated ion channel activity comparable to bladder SMCs which may be important for urological regenerative medicine applications.


Inferring the Genomic Landscape of Recombination Rate Variation in European Aspen (Populus tremula).
The rate of meiotic recombination is one of the central factors determining genome-wide levels of linkage disequilibrium which has important consequences for the efficiency of natural selection and for the dissection of quantitative traits. Here we present a new, high-resolution linkage map for Populus tremula that we use to anchor approximately two thirds of the P. tremula draft genome assembly on to the expected 19 chromosomes, providing us with the first chromosome-scale assembly for P. tremula (Table 2). We then use this resource to estimate variation in recombination rates across the P. tremula genome and compare these results to recombination rates based on linkage disequilibrium in a large number of unrelated individuals. We also assess how variation in recombination rates is associated with a number of genomic features, such as gene density, repeat density and methylation levels. We find that recombination rates obtained from the two methods largely agree, although the LD-based method identifies a number of genomic regions with very high recombination rates that the map-based method fails to detect. Linkage map and LD-based estimates of recombination rates are positively correlated and show similar correlations with other genomic features, showing that both methods can accurately infer recombination rate variation across the genome. Recombination rates are positively correlated with gene density and negatively correlated with repeat density and methylation levels, suggesting that recombination is largely directed toward gene regions in P. tremula.


The Role of Executive Function in Shaping Reinforcement Learning.
Reinforcement learning (RL) models have advanced our understanding of how animals learn and make decisions, and how the brain supports some aspects of learning. However, the neural computations that are explained by RL algorithms fall short of explaining many sophisticated aspects of human decision making, including the generalization of learned information, one-shot learning, and the synthesis of task information in complex environments.. Instead, these aspects of instrumental behavior are assumed to be supported by the brain's executive functions (EF). We review recent findings that highlight the importance of EF in learning. Specifically, we advance the theory that EF sets the stage for canonical RL computations in the brain, providing inputs that broaden their flexibility and applicability. Our theory has important implications for how to interpret RL computations in the brain and behavior.


Molecular differentiation of two sibling species of the black fly Simulium vittatum (Diptera: Simuliidae) based on random amplified polymorphic DNA.
Larvae of the black fly morphospecies Simulium vittatum from Colorado, Montana, Nebraska, and New Hampshire were cytologically identified as either the IS-7 or the IIIL-1 cytospecies. DNA was PCR amplified from cytotyped larvae using eight 10-mer primers, labeled with 33P, and run on polyacrylamide gels. The entire data set of 96 amplicons produced incomplete separation of the two cytospecies when subjected to neighbor-joining and maximum parsimony analyses. However, when analyzed within geographical regions, separate species status was supported. Bootstrap support for distinctness of the two cytospecies was best in Colorado where they were collected in true sympatry. The IS-7 cytospecies was more polymorphic in the western states, where it differed most from IIIL-1, which was most variable in the eastern states. The frequencies of the 17 most common amplicons in the two cytospecies were inversely correlated. A model of speciation derived from the molecular evidence suggests that IS-7 evolved in the west and spread eastward, whereas IIIL-1 later originated in the east and spread westward.


Mistimed sleep disrupts circadian regulation of the human transcriptome.
Circadian organization of the mammalian transcriptome is achieved by rhythmic recruitment of key modifiers of chromatin structure and transcriptional and translational processes. These rhythmic processes, together with posttranslational modification, constitute circadian oscillators in the brain and peripheral tissues, which drive rhythms in physiology and behavior, including the sleep-wake cycle. In humans, sleep is normally timed to occur during the biological night, when body temperature is low and melatonin is synthesized. Desynchrony of sleep-wake timing and other circadian rhythms, such as occurs in shift work and jet lag, is associated with disruption of rhythmicity in physiology and endocrinology. However, to what extent mistimed sleep affects the molecular regulators of circadian rhythmicity remains to be established. Here, we show that mistimed sleep leads to a reduction of rhythmic transcripts in the human blood transcriptome from 6.4% at baseline to 1.0% during forced desynchrony of sleep and centrally driven circadian rhythms. Transcripts affected are key regulators of gene expression, including those associated with chromatin modification (methylases and acetylases), transcription (RNA polymerase II), translation (ribosomal proteins, initiation, and elongation factors), temperature-regulated transcription (cold inducible RNA-binding proteins), and core clock genes including CLOCK and ARNTL (BMAL1). We also estimated the separate contribution of sleep and circadian rhythmicity and found that the sleep-wake cycle coordinates the timing of transcription and translation in particular. The data show that mistimed sleep affects molecular processes at the core of circadian rhythm generation and imply that appropriate timing of sleep contributes significantly to the overall temporal organization of the human transcriptome.


Effects of Lichenic Extracts (Hypogymnia physodes, Ramalina polymorpha and Usnea florida) on Human Blood Cells: Cytogenetic and Biochemical Study.
Several lichen species have been used for medicinal purposes throughout the ages, and they were reported to be effective in the treatment of different disorders including tuberculosis, hemorrhoids, ulcer, dysentery and cancer. It is revealed that they may be easily accessible sources of natural drugs that could be used as a possible food supplement or in pharmaceutical industry after their safety evaluations. However, so far, the nature and/or biological roles of plenty of lichenes have not been elucidated exactly. The aim of this study was to investigate the genetic and oxidative effects of water extracts of three different lichen species; Hypogymnia physodes, Ramalina polymorpha and Usnea florida in cultured human blood cells (n = 5) for the first time. All lichen species were collected from the Erzurum and Artvin provinces (in Turkey) during August 2010. The lichen extracts were added into culture tubes at various concentrations (0 to 2000 mg/L). Chromosome aberrations (CA) and micronucleus (MN) tests were used for genotoxic influences estimation. In addition, biochemical parameters (total antioxidant capacity (TAC) and total oxidative stress (TOS)) were examined to determine oxidative effects. In our in-vitro test systems, it was observed that all tested lichen extracts had no mutagenic effects on human lymphocytes. Furthermore, these extracts exhibited antioxidant properties due to the type of lichen species added to the cultures. In conclusion, these lichens can be a new resource of therapeutics as recognized in this study with their non-mutagenic and antioxidant features.


Gene and protein expression of O-GlcNAc-cycling enzymes in human laryngeal cancer.
Aberrant protein O-GlcNAcylation may contribute to the development and malignant behavior of many cancers. This modification is controlled by O-linked β-N-acetylglucosamine transferase (OGT) and O-GlcNAcase (OGA). The aim of this study was to determine the expression of O-GlcNAc cycling enzymes mRNA/protein and to investigate their relationship with clinicopathological parameters in laryngeal cancer. The mRNA levels of OGT and MGEA5 genes were determined in 106 squamous cell laryngeal cancer (SCLC) cases and 73 non-cancerous adjacent laryngeal mucosa (NCLM) controls using quantitative real-time PCR. The level of OGT and OGA proteins was analyzed by Western blot. A positive expression of OGT and MGEA5 transcripts and OGT and OGA proteins was confirmed in 75.5 and 68.9 % and in 43.7 and 59.4 % samples of SCLC, respectively. Higher levels of mRNA/protein for both OGT and OGA as well as significant increases of 60 % in total protein O-GlcNAcylation levels were noted in SCLC compared with NCLM (p < 0.05). As a result, an increased level of OGT and MGEA5 mRNA was related to larger tumor size, nodal metastases, higher grade and tumor behavior according to TFG scale, as well as incidence of disease recurrence (p < 0.05). An inverse association between OGT and MGEA5 transcripts was determined with regard to prognosis (p < 0.05). In addition, the highest OGT and OGA protein levels were observed in poorly differentiated tumors (p < 0.05). No correlations with other parameters were noted, but the results showed a trend of more advanced tumors to be more frequently OGT and OGA positive. The results suggest that increased O-GlcNAcylation may have an effect on tumor aggressiveness and prognosis in laryngeal cancer.


Liver X Receptor Genes Variants Modulate ALS Phenotype.
Amyotrophic lateral sclerosis (ALS) is one of the most severe motor neuron (MN) disorders in adults. Phenotype of ALS patients is highly variable and may be influenced by modulators of energy metabolism. Recent works have implicated the liver X receptors α and β (LXRs), either in the propagation process of ALS or in the maintenance of MN survival. LXRs are nuclear receptors activated by oxysterols, modulating cholesterol levels, a suspected modulator of ALS severity. In a cohort of 438 ALS patients and 330 healthy controls, the influence of LXR genes on ALS risk and phenotype was studied using single nucleotide polymorphisms (SNPs). The two LXRα SNPs rs2279238 and rs7120118 were shown to be associated with age at onset in ALS patients. Consistently, homozygotes were twice more correlated than were heterozygotes to delayed onset. The onset was thus delayed by 3.9 years for rs2279238 C/T carriers and 7.8 years for T/T carriers. Similar results were obtained for rs7120118 (+2.1 years and +6.7 years for T/C and C/C genotypes, respectively). The LXRβ SNP rs2695121 was also shown to be associated with a 30% increase of ALS duration (p = 0.0055, FDR = 0.044). The tested genotypes were not associated with ALS risk. These findings add further evidence to the suspected implication of LXR genes in the disease process of ALS and might open new perspectives in ALS therapeutics.


Cadmium Exposure as a Putative Risk Factor for the Development of Pancreatic Cancer: Three Different Lines of Evidence.
Although profoundly studied, etiology of pancreatic cancer (PC) is still rather scant. Exposure to cadmium (Cd), a ubiquitous metal associated with well-established toxic and carcinogenic properties, has been hypothesized to one putative cause of PC. Hence, we analyzed recently published observational studies, meta-analyses, and experimental animal and in vitro studies with the aim of summarizing the evidence of Cd involvement in PC development and describing the possible mechanisms. Consolidation of epidemiological data on PC and exposure to Cd indicated a significant association with an elevated risk of PC among general population exposed to Cd. Cadmium exposure of laboratory animals was showed to cause PC supporting the findings suggested by human studies. The concordance with human and animal studies is buttressed by in vitro studies, although in vitro data interpretation is problematic. In most instances, only significant effects are reported, and the concentrations of Cd are excessive, which would skew interpretation. Previous reports suggest that oxidative stress, apoptotic changes, and DNA cross-linking and hypermethylation are involved in Cd-mediated carcinogenesis. Undoubtedly, a significant amount of work is still needed to achieve a better understanding of the Cd involvement in pancreatic cancer which could facilitate prevention, diagnosis, and therapy of this fatal disease.


Interpreting patterns of gene expression with self-organizing maps: methods and application to hematopoietic differentiation.
Array technologies have made it straightforward to monitor simultaneously the expression pattern of thousands of genes. The challenge now is to interpret such massive data sets. The first step is to extract the fundamental patterns of gene expression inherent in the data. This paper describes the application of self-organizing maps, a type of mathematical cluster analysis that is particularly well suited for recognizing and classifying features in complex, multidimensional data. The method has been implemented in a publicly available computer package, GENECLUSTER, that performs the analytical calculations and provides easy data visualization. To illustrate the value of such analysis, the approach is applied to hematopoietic differentiation in four well studied models (HL-60, U937, Jurkat, and NB4 cells). Expression patterns of some 6,000 human genes were assayed, and an online database was created. GENECLUSTER was used to organize the genes into biologically relevant clusters that suggest novel hypotheses about hematopoietic differentiation-for example, highlighting certain genes and pathways involved in "differentiation therapy" used in the treatment of acute promyelocytic leukemia.


Scalar and vector holographic gratings recorded in a photoanisotropic amorphous As(2)S(3) thin film.
Theoretical explanations of the kinetics of scalar and vector holographic gratings are developed, and experimental results are obtained in amorphous AS(2)S(3) thin films. The large nonlinear-optical susceptibilities are evaluated from experimental data in the vicinity of the fundamental absorption edge, and it is found that chi((3))(1111) congruent with 3.2 x 10(-2) esu and chi((3))(1122)/chi((3))(1111) congruent with 0.98.


Genome-wide association studies in East Asians identify new loci for waist-hip ratio and waist circumference.
Sixty genetic loci associated with abdominal obesity, measured by waist circumference (WC) and waist-hip ratio (WHR), have been previously identified, primarily from studies conducted in European-ancestry populations. We conducted a meta-analysis of associations of abdominal obesity with approximately 2.5 million single nucleotide polymorphisms (SNPs) among 53,052 (for WC) and 48,312 (for WHR) individuals of Asian descent, and replicated 33 selected SNPs among 3,762 to 17,110 additional individuals. We identified four novel loci near the EFEMP1, ADAMTSL3 , CNPY2, and GNAS genes that were associated with WC after adjustment for body mass index (BMI); two loci near the NID2 and HLA-DRB5 genes associated with WHR after adjustment for BMI, and three loci near the CEP120, TSC22D2, and SLC22A2 genes associated with WC without adjustment for BMI. Functional enrichment analyses revealed enrichment of corticotropin-releasing hormone signaling, GNRH signaling, and/or CDK5 signaling pathways for those newly-identified loci. Our study provides additional insight on genetic contribution to abdominal obesity.


Stabilization of Z DNA in vivo by localized supercoiling.
Biological processes such as transcription may generate domains of supercoiling on a circular DNA. The existence of these domains in Escherichia coli was investigated by the ability of different lengths of (CG) tracts, cloned upstream or downstream from the tetracycline resistance gene (tet) of pBR322, to adopt the Z structure in vivo. Segments as short as 12 base pairs adopt the Z form when cloned upstream from the tet gene (Eco RI site), whereas no Z DNA was detected when this sequence was cloned downstream (Sty I site), even with a 74-base pair (CG) tract that requires less supercoiling than shorter tracts for the B-Z transition. Hence the localized supercoil density in pBR322 can be as high as -0.038 and as low as -0.021 at different loci. These data demonstrate the existence of the Z structure for commonly found natural sequences and support the notion of domains of negative supercoiling in vivo.


Modulation of proteoglycan metabolism by aortic smooth muscle cells grown on collagen gels.
The nature of the extracellular matrix may influence the types and amounts of proteoglycans synthesized by arterial smooth muscle cells. To determine if collagen modulates proteoglycan metabolism by these cells, arterial smooth muscle cells derived from aortic explants of the pigtail monkey (Macaca nemestrina) were cultured on both tissue culture plastic and hydrated type I collagen gels for 7 days. Cells were radiolabeled with Na2[35S]O4 during the final 48 hours of the growth period, and proteoglycans were extracted from the culture medium and cell layer by using 4 M guanidine hydrochloride in the presence of protease inhibitors. Cultures on collagen accumulated approximately 40% less [35S]O4 = -radiolabeled proteoglycan when compared to cultures on plastic. However, cells on collagen accumulated approximately 50% of their newly synthesized proteoglycan within the cell layer, while cells on plastic deposited less than 20% of their total radiolabeled proteoglycan in this culture compartment. Pulse-chase analysis indicated that the elevated accumulation of proteoglycan observed in the cell layer of collagen cultures was due, at least in part, to inhibition of turnover of these molecules. Cells on both collagen and plastic synthesized a large chondroitin sulfate proteoglycan which was secreted into the medium and deposited within the cell layer. On the other hand, cells on collagen synthesized a smaller iduronic acid-rich dermatan sulfate proteoglycan which was deposited only within the collagen gel and not secreted into the medium. By comparison, cells grown on plastic synthesized both a small glucuronic acid-rich dermatan sulfate proteoglycan which was secreted into the medium as well as an iduronic acid-rich dermatan sulfate which was present in the cell layer. Unlike the cell layer-derived dermatan sulfate proteoglycan isolated from the collagen gels, the majority of the cell layer-derived dermatan sulfate from cells on plastic was insensitive to papain treatment and thus identified as free glycosaminoglycan chains. Analysis of the total radiolabeled proteoglycans isolated under the two culture conditions indicated that cultures grown on collagen accumulate less chondroitin sulfate proteoglycan and heparan sulfate proteoglycan but over twice as much iduronic acid-rich dermatan sulfate proteoglycan. This culture system is offered as a model to determine the mechanisms by which collagen may in part regulate the metabolism of proteoglycans by arterial smooth muscle cells.