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Loop mediated isothermal amplification (LAMP) is an innovative technique which allows the rapid detection of target nucleic acid sequences under isothermal conditions without the need for complex instrumentation. The development, systematic optimisation and clinical validation of a LAMP assay targeting the ctrA gene for the rapid detect ion of capsular N. meningitidis is described. Highly specific detection of capsular N. meningitidis type strains and clinical isolates was demonstrated, with no cross reactivity with other Neisseria spp., or with a comprehensive panel of other common human pathogens. The lower limit of detection was determined as 6 ctrA gene copies detectable in 48 minutes, with positive reactions readily identifiable visually via a simple colour change. High copy numbers could be detected in 22 minutes. Clinical Validation was carried out by retrospectively and prospectively analysing a total of 935 clinical specimens from a total of 603 patients. The level of agreement between the ctrA LAMP assay and a confirmatory test result (Culture, MRU PCR, ctrA RT-PCR) for each patient was used to evaluate the performance of LAMP for the detection of capsular N. meningitidis. The overall sensitivity and specificity of the designed ctrA LAMP assay was determined as 94.6% and 99.6% with positive and negative predictive values of 96.4% and 99.5% respectively. The potential utility of a positive ctrA LAMP respiratory specimen result to indicate MD was investigated. The sensitivity and specificity of respiratory• specimens in this regard was 89.3% and 97.7% with positive and negative predictive values of 80.7% and 98.9% respectively. Rapid LAMP testing of non-invasive respiratory specimens has significant potential to accelerate the diagnosis of MD. The LAMP method represents a simple, rapid, sensitive, and highly specific technique for the detection of N. meningitidis, and has the potential to be used as a POC molecular test and in resource poor settings.

The work presented in this thesis was undertaken to increase understanding of the intracellular mechanisms regulating acid secretion by gastric parietal cells. Investigation of the effects of protein kinase C on secretory activity induced by a variety of agents was a major objective. A further aim was to establish the sites at which epidermal growth factor (EGF) acts to stimulate prostaglandin E2 (PGE2) production and to inhibit acid secretion. These investigations were carried out by using the HGT-1 human gastric cancer cell line and freshly isolated rat parietal cells. In HGT-1 cells, the cyclic AMP response to histamine and to truncated glucagon-like peptide 1 (TGLP-1) was reduced when protein kinase C was activated by 12-0-tetradecanoylphorbol 13-acetate (TPA). Receptor-binding studies and experiments in which cyclic AMP production in HGT-1 cells was stimulated by gastric inhibitory polypeptide, cholera toxin and forskolin suggested that the effect of TPA was mediated by uncoupling of the histamine H2 receptor from the guanine nucleotide regulatory protein Gs, possibly by phosphorylation of the receptor. An involvement of protein kinase C α in this effect was suggested because an antibody to this isoform specifically prevented the inhibitory effects of TPA on histamine-stimulated adenylate cyclase activity in a membrane fraction prepared from HGT-1 cells. Carbachol-stimulated secretory activity in parietal cells was specifically inhibited by Ro 31-8220, a bisindolylmaleimide inhibitor of protein kinase C. Thus protein kinase C may play a role in the activation of the secretory response to carbachol. In parietal cells prelabelled with [3H]-arachidonic acid or [3H]myristic acid, EGF did not affect [3H]-fatty acid or [3H] - diacylglycerol content. No evidence for effects of EGF on phosphatidylinositol glycan-specific phospholipase C, phospholipase A2 or on low Km cyclic AMP phosphodiesterase activities were found.

Previous work has shown D1/D2 requisite synergism can still occur in the striatum in the absence of action potentials. Some nonclassical communication such as gap junctions may be allowing the segregated dopamine (DA) receptors to interact to produce stereotyped motor activity. Connexin-32 (Cx32) and connexin-36 (Cx36) were targeted for study due to their abundance in neural tissues and presence in the striatum. Mice lacking either the Cx32 or Cx36 gene and their respective wildtype littermates were compared on a climbing behavior task used to gauge their dopaminergic activity after receiving either saline, D1 agonist, D2 agonist, or both D1 and D2 agonists. The results showed that D1/D2 requisite synergism was still intact in both strains of mice. The Cx32 WT mice displayed significantly greater scores than the KO mice in the D1/D2 treatment. The Cx36 mice did not display a significant genotype difference, but a trend was observed with the KO females having larger scores relative to WT females or to males of either genotype.

Cool-season perennial grass growth is suppressed during hot, dry summers. This "summer slump" reduces the availability of grazable forage. Tall-growing perennial warm-season grasses that produce 65 to 75% of their yield in midsummer may provide needed herbage during this period of reduced cool-season forage production. However, establishment of perennial warm-season species is slow and inconsistent when compared to cool-season species. Before these warm-season species can be relied on for summer forage production, a successful establishment methodology must be developed. A study was conducted to evaluate the influence of atrazine [2-chloro·4(ethylarnino)-s-triazine] and carbofuran on establishment of switchgrass (Panicum virgatum L.) and Caucasian bluestem [Boyhtiochloa caucasica (Trin.) C. E. Hubbard]. Treatments of carbofuran at 0 and 1.1 atrazine/ha placed in the row with the seed and of atrazine broadcast at 0, 1.1, and 2.2 kg/ha were imposed in all possible combinations. Another study investigated the influence of limestone, P, and carbofuran on the establishment of switchgrass. Treatments included carbofuran at 0 and 1.1 kg/ha, limestone at 0 and 4.48 Mg/ha, and P at 0 and 22 kg/ha in all possible combinations. Seedling growth rate, leaf appearance rate, plant height, and leaf elongation rate (LER) of seedlings were recorded. Seedling weight and populations were determined at the sixth leaf stage of development. Yields of forage and botanical compositions were also measured in the establishment year. Establishment was further evaluated with yield measurements the year after seeding. Carbofuran application increased first year yield, seedling weight, population, leaf appearance rate, and seedling growth rate. Atrazine reduced seedling population, weight, leaf appearance rate, LER, and yield. Carbofuran partially moderated atrazine influence. The 2.2 kg rate of atrazine tended to injure the switchgrass more than the Caucasian bluestem. In the study investigating limestone and P, carbofuran increased yield, seedling weight, seedling population, LER, seedling growth rate, and leaf appearance rate. The greatest yield, leaf appearance rate, and LER occurred with applications ofP and carbofuran. Even though initial soil pH was 5.1, limestone broadcast prior to planting did not influence seedling measurements and yields in either year. These plantings were made during 1985 and 1986, which were the driest summers in recent years. In spite of the moisture stress, acceptable stands of perennial warm·season grasses were established in both experiments in both years. Including 1.1 kg granular carbofuran/ha in the row, and broadcast application of 1.1 kg atrazine/ha at planting will improve establishment of perennial warm·season grasses. Where the P levels in the soil are low, a broadcast application of 22 kg P/ha improves chances for successful establishment.

InDecor Ltd., Inc. is a small manufacturer of the Avant series of candleholders and is located in Omaha, Nebraska. Annual sales have increased from $20,400 in 1969 when the company was founded to $72,000 in 1972. | Manufacturing is of the sheet metal fabrication class and involves light punch press operations, assembly, welding, finishing, and packaging. | The company distributes some 59 styles of candleholders and associated starter groupings (Exhibit 5) solely through factory coordinated sales representatives. These representatives have exclusive rights within their respective regions and display the products at regional gift shows. A large portion of their sales is derived from calling directly on retailers and each representative receives a 20% commission on gross sales. All shipments and invoicing and any special problems are handled by the factory directly with the customer. Sales are also stimulated through a continuous advertising program in the two prime trade magazines and by the direct mailing of catalogs with self-addressed order forms to past customers.

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4: 198-205, 1939