Cloning, mapping and identification of the Aeromonas salmonicida fstA, fepA and irpA genes, encoding the 86, 82 and 74 kDa iron-regulated outer membrane proteins, respectively

Three iromps (iron-regulated outer membrane proteins) of Aeromonas salmonicida were identified by the use of specific antibodies together with Southern hybridization analysis and limited nucleotide sequencing of their genes. The results of these experiments together with a search of the international database for homologous sequences led to their identification as follows: -86 kDa iromp (FstA) as a Vibrio anguillarum Fat A homologue -82 kDa iromp (FepA) as an Escherichia coli FepA homologue -74 kDa iromp (IrpA) as an Escherichia coli Cir homologue.

Immunostimulatory effects of different aspects of aquaculture on the host response in the edible sea urchin, paracentrotus lividus

Aquaculture is a fast-growing industry contributing to global food security and sustainable aquaculture, which may reduce pressures on capture fisheries. The overall objective of this thesis was to look at the immunostimulatory effects of different aspects of aquaculture on the host response of the edible sea urchin, Paracentrotus lividus, which are a prized delicacy (roe) in many Asian and Mediterranean countries. In Chapter 1, the importance of understanding the biology, ecology, and physiology of P. lividus, as well as the current status in the culture of this organism for mass production and introducing the thesis objectives for following chapters is discussed. As the research commenced, the difficulties of identifying individuals for repeat sampling became clear; therefore, Chapter 2 was a tagging experiment that indicated PIT tagging was a successful way of identifying individual sea urchins over time with a high tag retention rate. However, it was also found that repeat sampling via syringe to measure host response of an individual caused stress which masked results and thus animals would be sampled and sacrificed going forward. Additionally, from personal observations and discussion with peers, it was suggested to look at the effect that diet has on sea urchin immune function and the parameters I measured which led to Chapter 3. In this chapter, both Laminaria digitata and Mytilus edulis were shown to influence measured immune parameters of differential cell counts, nitric oxide production, and lysozyme activity. Therefore, trials commencing after Trial 5 in Chapter 4, were modified to include starvation in order to remove any effect of diet. Another important aspect of culturing any organism is the study of their immune function and its response to several immunostimulatory agents (Chapter 4). Zymosan A was shown to be an effective immunostimulatory agent in P. lividus. Further work on handled/stored animals (Chapter 5) showed Zymosan A reduced the measured levels of some immune parameters measured relative to the control, which may reduce the amount of stress in the animals. In Chapter 6, animals were infected with Vibrio anguillarum and, although V. anguillarum, impacted immune parameters of P. lividus, it did not cause mortality as predicted. Lastly, throughout this thesis work, it was noted that the immune parameters measured produced different values at different times of the year (Chapter 7); therefore, using collated baseline (control) data, results were compiled to observe seasonal effects. It was determined that both seasonality and sourcing sites influenced immune parameter measurements taken at different times throughout the year. In conclusion, this thesis work fits into the framework of development of aquaculture practices that affect immune function of the host and future research focusing on the edible sea urchin, P. lividus.

Isolation of a pathogenic strain of Vibrio alginolyticus from necrotic larvae of Macrobrachium rosenbergii (de Man)

Giant freshwater prawn, Macrobrachium rosenbergii (de Man), is an important commercial species with considerable export value, ideal for cultivation under low saline conditions and in freshwater zones (Kurup 1994). However, despite more than a decade of research on its larval production systems, vibriosis still hampers seed production resulting in high mortality rates. Among the different species of vibrios, Vibrio alginolyticus has been isolated frequently from diseased shrimp as the aetiological agent of vibriosis and has been described as a principal pathogen of both penaeids and nonpenaeids (Lightner 1988; Baticados, Cruz-Lacierda, de la Cruz, Duremdez-Fernandez, Gacutan, Lavilla- Pitogo & Lio-Po 1990; Mohney, Lightner & Bell 1994; Lee, Yu, Chen, Yang & Liu 1996). Vibrio fluvialis, V. alginolyticus, V. cholerae non-O1 (Fujioka & Greco 1984), Aeromonas liquifaciens and V. anguillarum (Colorni 1985) have been isolated from the larvae of M. rosenbergii. A profound relationship between the abundance of members of the family Vibrionaceae and larval mortality (Singh 1990) and the predominance of Vibrio in eggs, larvae and post-larvae of M. rosenbergii (Hameed, Rahaman, Alagan & Yoganandhan 2003) was reported. The present paper reports the isolation, characterization, pathogenicity and antibiotic sensitivity of V. alginolyticus associated with M. rosenbergii larvae during an occurrence of severe mass mortality at the ninth larval stage.

Alkaline protease from a non-toxigenic mangrove isolate of Vibrio sp. V26 with potential application in animal cell culture

Vibrio sp. V26 isolated from mangrove sediment showed 98 % similarity to 16S rRNA gene of Vibrio cholerae, V. mimicus, V. albensis and uncultured clones of Vibrio. Phenotypically also it resembled both V. cholerae and V. mimicus.Serogrouping, virulence associated gene profiling, hydrophobicity, and adherence pattern clearly pointed towards the non—toxigenic nature of Vibrio sp. V26. Purification and characterization of the enzyme revealed that it was moderately thermoactive, nonhemagglutinating alkaline metalloprotease with a molecular mass of 32 kDa. The application of alkaline protease from Vibrio sp. V26 (APV26) in sub culturing cell lines (HEp-2, HeLa and RTG-2) and dissociation of animal tissue (chick embryo) for primary cell culture were investigated. The time required for dissociation of cells as well as the viable cell yield obtained by while administeringAPV26 and trypsin were compared. Investigations revealed that the alkaline protease of Vibrio sp. V26 has the potential to be used in animal cell culture for subculturing cell lines and dissociation of animal tissue for the development of primary cell cultures, which has not been reported earlier among metalloproteases of Vibrios.

Development of monoclonal antibodies against Vibrio pathogens

Monoclonal antibodies were developed against pathogenic vibrios for use in rapid identification in disease situations of humans, fish and shellfish. Of the 12 fusions performed using V. alginolyticus, V. anguillarum, V. carchariae, V. cholerae, V. damsela, V. furnissii, V. harveyi, V. ordalii, V. parahaemolyticus and V. vulnificus, a total of 102 hybridomas were obtained. Based on cross-reactivity of a wide range of Vibrio strains and other gram-negative bacteria, three broad types of monoclonal antibodies were found. The three categories were: (1) ones that were species-specific or specific to a particular surface antigen, (2) a large number that reacted with several Vibrio species, and (3) three that reacted with most Vibrio strains but no other gram-negative bacteria. Each species-specific monoclonal antibody only recognized its corresponding Vibrio species and was used for identifying unknown species, confirming diagnosis of clinical isolates. In addition, several monoclonal antibodies only cross-reacted with similar Vibrio species, e.g. V. parahaemolyticus and V. alginolyticus which share a common H-antigen. Monoclonal antibodies reacting with several Vibrio species were not of particular use in diagnostic situations. Three monoclonal antibodies of the last group did not react with other genera of the family Vibrionaceae, namely Aeromonas, Photobacterium and Plesiomonas nor a wide range of gram-negative enteric bacteria. These data indicated the existence of an antigenic surface determinant common to Vibrio species. One monoclonal reacted with the heat-stable antigenic determinants on the cell surface as v as lipopolysaccharide extracted from all the vibrios studied, thus making it useful for large- scale screening of acute infections of vibrios. In a blind test, seven Vibrio species, isolated from 6 marine and a freshwater source were identified by two laboratories using phenetic tests. Results of immunotyping using monoclonals, three of seven were diagnosed as the same species, another three were designated as Vibrio species but could not be classified further due to the library not having the corresponding monoclonal, and one was diagnostically questionable. Two further tests were carried out. An unknown Vibrio formalin-fixed isolated from diseased marine animal was identified as V. parahaemolyticus by ELISA and FITC. Clinical human isolates of V. alginolyticus, V. parahaemolyticus and V. vulnificus were confirmed by monoclonals. Australian isolates of V. anguillarum appeared to be mostly of serotype O1. monoclonals raised to V. anguillarum AFHRL 1 reacted with only serotype O1 from Denmark but also most Australian isolates. All vibrios pathogenic to fish and shellfish, i.e. V. anguillarum, V. ordalii, V. alginolyticus, V. carchariae, V. cholerae, V. damsela, V. harveyi, V. parahaemolyticus and V. vulnificus, were used for attachment studies to fish cells using phase contrast and FITC-immunofluorescence microscopy. Of these vibrios, V. anguillarum, V. ordalii and V. perahaemolyticus, were found to adhere to different cells and tissues of rainbow trout while others did not appear to attach. However, attachment was inhibited by monoclonal antibodies specific to only these three vibrios. Lipopolysaccharide is well known as being a contributing factor in pathogenicity of gram-negative bacteria. PAGE electrophoresis of extracted LPS from 9 strains covering 6 Vibrio species showed the presence of a common 15,000 D fragment. This fragment was verified by immunoblotting with a genus-specific monoclonal antibody (i.e. F11P411F) recognizing nearly all vibrios. The common LPS fragment was separated and used to raise polyclonal antisera in mouse which reacted strongly with LPS itself, live as well as sodium azide-killed vibrios, but not with other gram-negative bacteria. This raised the possibility of developing vaccine from Vibrio LPS. Monoclonal antibodies developed in the present study enabled rapid identification of a number of pathogenic Vibrio species. There is still further work to produce monoclonal antibodies against additional vibrios that are probably pathogenic. These included V. fluvialis, V. hollisae, V. metschnikovii, V. minicus, V. salmonella and V. tubiashii. Together the application will be of significance in clinical diagnostic work, in the monitoring of vibriosis in fish farms and in quarantine.

A comparison of the hazard perception ability of matched groups of healthy drivers aged 35 to 55, 65 to 74, and 75 to 84 years

We examined differences in response latencies obtained during a validated video-based hazard perception driving test between three healthy, community-dwelling groups: 22 mid-aged (35-55 years), 34 young-old (65-74 years), and 23 old-old (75-84 years) current drivers, matched for gender, education level, and vocabulary. We found no significant difference in performance between mid-aged and young-old groups, but the old-old group was significantly slower than the other two groups. The differences between the old-old group and the other groups combined were independently mediated by useful field of view (UFOV), contrast sensitivity, and simple reaction time measures. Given that hazard perception latency has been linked with increased crash risk, these results are consistent with the idea that increased crash risk in older adults could be a function of poorer hazard perception, though this decline does not appear to manifest until age 75+ in healthy drivers.

Railways, Urban Development and Town Planning in Britain: 1948–2008, Haywood R. Ashgate (2009). £65.00, ISBN: 9780754673927

This book is a thorough investigation of the relationship between land use planning and the railways in Britain, through review of the factors affecting the two sectors and their integration during the period of public ownership. The rationale behind the book is explained as a timely analysis of the dynamic correlation involving town planning and management of the railway in a period when growing congestion on the road network is forcing people to look for alternative modes and capacity is badly needed to accommodate this increased demand for travel. The book calls for a modal shift from road to rail for passenger and freight traffic.

Nutrition screening and risk factors prior to chemotherapy in cancer patients 65 years and older

Background The largest proportion of cancer patients are aged 65 years and over. Increasing age is also associated with nutritional risk and multi-morbidities—factors which complicate the cancer treatment decision-making process in older patients. Objectives To determine whether malnutrition risk and Body Mass Index (BMI) are associated with key oncogeriatric variables as potential predictors of chemotherapy outcomes in geriatric oncology patients with solid tumours. Methods In this longitudinal study, geriatric oncology patients (aged ≥65 years) received a Comprehensive Geriatric Assessment (CGA) for baseline data collection prior to the commencement of chemotherapy treatment. Malnutrition risk was assessed using the Malnutrition Screening Tool (MST) and BMI was calculated using anthropometric data. Nutritional risk was compared with other variables collected as part of standard CGA. Associations were determined by chi-square tests and correlations. Results Over half of the 175 geriatric oncology patients were at risk of malnutrition (53.1%) according to MST. BMI ranged from 15.5–50.9kg/m2, with 35.4% of the cohort overweight when compared to geriatric cutoffs. Malnutrition risk was more prevalent in those who were underweight (70%) although many overweight participants presented as at risk (34%). Malnutrition risk was associated with a diagnosis of colorectal or lung cancer (p=0.001), dependence in activities of daily living (p=0.015) and impaired cognition (p=0.049). Malnutrition risk was positively associated with vulnerability to intensive cancer therapy (rho=0.16, p=0.038). Larger BMI was associated with a greater number of multi-morbidities (rho =.27, p=0.001. Conclusions Malnutrition risk is prevalent among geriatric patients undergoing chemotherapy, is more common in colorectal and lung cancer diagnoses, is associated with impaired functionality and cognition and negatively influences ability to complete planned intensive chemotherapy.

Health Information Literacy and the Experience of 65 to 79 Year Old Australians

Information literacy is presented here from a relational perspective, as people’s experience of using information to learn in a particular context. A detailed practical example of such a context is provided, in the health information literacy experience of 65–79 year old Australians. A phenomenographic investigation found five qualitatively distinct ways of experiencing health information literacy: Absorbing (intuitive reception), Targeting (a planned process), Journeying (a personal quest), Liberating (equipping for independence) and Collaborating (interacting in community). These five ways of experiencing indicated expanding awareness of context (degree of orientation towards their environment), source (breadth of esteemed information), beneficiary (the scope of people who gain) and agency (amount of activity), across HIL core aspects of information, learning and health. These results illustrate the potential contribution of relational information literacy to information science.

Comparative studies of how living circumstances influence medication adherence in ≥ 65 year olds

Background Resources to help the older aged (≥65 year olds) manage their medicines should probably target those in greatest need. The older-aged have many different types of living circumstances. There are different locations (urban, rural), different types of housing (in the community or in retirement villages), different living arrangements (living alone or with others), and different socioeconomic status (SES) circumstances. However, there has been limited attention to whether these living circumstances affect adherence to medicines in the ≥65 year olds. Aim of the review The aim was to determine whether comparative studies, including logistic regression studies, show that living circumstances affect adherence to medicines by the ≥65 year olds. Methods A literature search of Medline, CINAHL and the Internet (Google) was undertaken. Results Four comparative studies have not shown differences in adherence to medicines between the ≥65 year olds living in rural and urban locations, but one study shows lower adherence to medicines for osteoporosis in rural areas compared to metropolitan, and another study shows greater adherence to antihypertensive medicines in rural than urban areas. There are no comparative studies of adherence to medicines in the older-aged living in indigenous communities compared to other communities. There is conflicting evidence as to whether living alone, being unmarried, or having a low income/worth is associated with nonadherence. Preliminary studies have suggested that the older-aged living in rental, low SES retirement villages or leasehold, middle SES retirement villages have a lower adherence to medicines than those living in freehold, high SES retirement villages. Conclusions The ≥65 year olds living in rural communities may need extra help with adherence to medicines for osteoporosis. The ≥65 year olds living in rental or leasehold retirement villages may require extra assistance/resources to adhere to their medicines. Further research is needed to clarify whether living under certain living circumstances (e.g. living alone, being unmarried, low income) has an effect on adherence, and to determine whether the ≥65 year olds living in indigenous communities need assistance to be adherent to prescribed medicines.

Plan Krivorogskogo okruga Bozhedarovskogo raĭona na uchastok zemli kolfonda N 65 poselok N 3

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Process variability-aware statistical hybrid modeling of dynamic power dissipation in 65 nm CMOS designs

A generalized technique is proposed for modeling the effects of process variations on dynamic power by directly relating the variations in process parameters to variations in dynamic power of a digital circuit. The dynamic power of a 2-input NAND gate is characterized by mixed-mode simulations, to be used as a library element for 65mn gate length technology. The proposed methodology is demonstrated with a multiplier circuit built using the NAND gate library, by characterizing its dynamic power through Monte Carlo analysis. The statistical technique of Response. Surface Methodology (RSM) using Design of Experiments (DOE) and Least Squares Method (LSM), are employed to generate a "hybrid model" for gate power to account for simultaneous variations in multiple process parameters. We demonstrate that our hybrid model based statistical design approach results in considerable savings in the power budget of low power CMOS designs with an error of less than 1%, with significant reductions in uncertainty by atleast 6X on a normalized basis, against worst case design.

The characteristics and questionable taxonomic position of the oxalate-decomposing bacterium, Vibrio extorquens

A detailed account of the morphological, staining, penicillin sensitivity and serological peculiarities of five strains of an oxalate-decomposing bacterium including the well-recognized strain Vibrio extorquens, has been given. Inasmuch as all the strains share many of the characteristics of the genus Arthrobacter and not Vibrio the desirability of placing the bacterium in the former genus for the time being has been suggested. The possibility of the strains falling under an altogether new genus which represent a phylogenic link between the pseudomonads and diphtheroids has been speculated.

Työhön liittyvä kilpailu 18 - 65-vuotiaiden pyhtääläisten kuvaamana

Tutkimuksen kohteena olivat Pyhtään työikäisten asukkaiden näkemykset ja kokemukset työssä esiintyvästä kilpailusta Tutkimusongelma oli työhön liittyvän kilpailun ilmiön hahmottaminen. Tutkimuksen tarkoitus oli valottaa työhön liittyvän kilpailun moninaisuutta antaen esimerkkejä sen eri muodoista, esittäen ajatuksia uusien tulkintojen tekemiselle ja herättäen ideoita kilpailun eri muotojen soveltamiselle työn ja työilmapiirin tutkimuksissa. Tavoitteena oli tutkia haastatteluissa esiintyviä tapoja käsittää työhön liittyvä kilpailu ja sitä, missä määrin eri ammateissa kuvattiin sosiaalista kilpailua verrattuna välineelliseen kilpailuun. Työyhteisöön samastumista tarkasteltiin kilpailun ja koetun oikeudenmukaisuuden näkökulmista. Työn arvostusta käsiteltiin tarkastellen haastateltujen käsitystä ammattinsa yleisestä arvostuksesta sekä haastateltujen omia arvioita ammatistaan, sekä millä tavalla ammattirooli nousi esiin haastateltujen itsen kuvauksissa. Työpaikan ihmissuhteita ja valtaa tarkasteltiin kohdistamalla huomio työtovereiden kanssa toimeen tulemiseen ja työpaikan ihmissuhteiden aiheuttamaan stressiin sekä haastateltujen työssään kokeman oman vaikutusvallan, tasavertaisuuden, epäreiluuden, arvostuksen ja kilpailun määrään. Selvitettiin myös miten vallankäyttöä ja kiusaamista kuvattiin ja millä tavalla vallankäytön ja kiusaamisen katsottiin edesauttavan tavoitteiden saavuttamisessa. Lopuksi tarkastelun kohteina olivat ongelmien käsittely, työssä viihtyminen ja viihtymiseen vaikuttavat seikat. Aineisto kerättiin Pyhtäällä kesällä 2007. Vastaajia oli yhteensä 245 ja vastausprosentti oli 50. Tutkimuksessa mukana olivat kalkki työilmapiiriä ja työpaikan ihmissuhteita koskeviin kysymyksiin vastanneet haastatellut (N = 167). Suurimmaksi osaksi tutkimus oli kuvailevaa, mutta siinä oli myös kvantitatiivista kysymyksenasettelua. Tutkimuksessa käytettiin sekä kvalitatiivisia että kvantitatiivisia analyysimenetelmiä. Puolistrukturoiduissa yksilöhaastatteluissa hyödynnettiin tulkitsevaa fenomenologista analyysia. Kirjattuja avovastauksia tutkittiin luokittavalla ja tulkitsevalla sisällönanalyysilla. Lisäksi laskettiin tilastollisia merkitsevyyksiä Työpaikalla esiintyvä kilpailu nähtiin haastateltujen selonteoissa hyvin moninaisena, sekä myönteisenä että haitallisena. Siihen ymmärrettiin kuuluvan niin resursseihin, työtehtäviin, tuloksiin ja palkkioihin liittyvää välineellistä kilpailua, kuin ihmisten välistä sosiaalista valtataisteluakin. Suhteessa välineelliseen kilpailuun sosiaalista kilpailua kuvattiin eniten hoiva- ja sosiaalialan ammateissa. Suhteessa sosiaaliseen kilpailuun välineellistä kilpailua kuvasivat eniten yrittäjät Vastausten perusteella suun kilpailun määrä korreloi työpaikan ihmissuhteiden aiheuttamaan stressiin, mutta vain silloin, kun kokemus työpaikan oikeudenmukaisuudesta oli matala. Kilpailun eri muotojen (kilpailu omien tavoitteiden saavuttamisesta vs. kilpailu organisaation tavoitteiden saavuttamisesta) väliset yhteydet työyhteisöön samastumiseen eivät aineistossa olleet tilastollisesti merkitseviä. Haastatellun arvio oman ammattinsa arvostuksesta ei myöskään ennustanut haastatellun todennäköisyyttä esittää itsensä ammattikuntansa edustajana avoimen itsen kuvauksen tilanteessa. Koettu oikeudenmukaisuus kuitenkin korreloi työyhteisöön samastumiseen. Mitä oikeudenmukaisempana haastatellut työpaikkaansa pitivät, sitä tärkeämmäksi samastumiskohteeksi he sen tyypillisesti arvioivat. Myös työssä viihtymisen ja työpaikan koetun oikeudenmukaisuuden välinen korrelaatio oli aineistossa voimakas. Lisäksi silloin, kun kokemus oikeudenmukaisuudesta oli matalalla tasolla, oli myös työpaikan ihmissuhteiden aiheuttama stressi yhteydessä vähäiseen työssä viihtymiseen. Tärkeimmät läheet tutkimuksessa olivat: Turner. John C 1975. Social comparison and social identity. Some prospects for intergroup behaviour. European Journal of Social Psychology, 5(1), 5-34. Haslam, S. Alexander 2004. Psychology in organizalions. The social identity approach Second Edition. London: SAGE Publications. Tyler, Tom - Blader. Steven 2000. Cooperation in Groups: Procedural Justice, Social Identity, and Behavioural Engagement. Essays in Social Psychology. Philadelphia: Psychology Press / Taylor & Francis Group. Kantolahti, T. - Tikander, T. 2010. Puheenvuoroja työn kuormittavuudesta. Sosiaali- ja terveysministeriön selvityksiä 2010:17

Studies of interfaces in Al-65 Cu20Fe15

The study of interfaces in quasicrystalline alloys is relatively new. Apart From the change in orientation, symmetry and chemistry which can occur across homophase and heterophase boundaries in crystalline materials, we have the additional, exciting possibility of an interface between quasicrystalline and its rational approximant. High resolution electron microscopy is a powerful technique to study the structural details of such interfaces. We report the results of a HREM study of the interface between the icosahedral phase and the related Al13Fe4 type monoclinic phase in melt spun and annealed Al65Cu20Fe15 alloy.