Dissertations / Theses on the topic 'Preservative efficacy'

The preservative challenge test is a regulatory requirement specified in various pharmacopoeias to determine the efficacy of preservatives. However, such testing is a labour-intensive repetitive task and often requires days before results can be generated. Microbial biosensors have the potential to provide a rapid and automated alternative to the traditional viable counting currently in use. However, the selection of appropriate promoters is essential. The bioluminescent reporter strains used in the current study comprise the Photorhabdus luminescence lux CDABE reporter genes under the control of five individual constitutive Escherichia coli promoters: outer lipoprotein (lpp); twin arginine translocase (tatA); lysine decarboxylase (ldc); lysyl t-RNA (lysS); and ribosomal protein (spc). The promoter plus lux CDABE constructs were cloned, ligated into the plasmid vector pBR322 and transformed into E. coli ATCC 8739. The bioluminescence intensity in the decreasing order of constitutive promoter was lpp > spc> tatA> ldc > lysS. The five biosensor strains tested successfully in PET assays and demonstrated accuracy with a minimum detection limit of 103 CFU/ml, a detection range of 6 orders magnitude, and yielded equivalent results to methods currently recommended by the pharmacopoeias. The bioluminescent biosensors were used to monitor the efficacy of preservatives; sorbic acid at concentrations of 0.031% to 0.2% at pH 5.0, and benzalkonium chloride at concentrations of 0.0062% to 0.00039% alone and in combination with 0.03% EDTA. The 99.9% percentage of bioluminescence reduction of tatA-lux, ldc-lux, lysS-lux, and spc-lux was statistically equivalent to the 3 log10 CFU/ml reduction as required by the Pharmacopeias’. Strong significant correlations between bioluminescence and the methods recommended by the pharmacopoeias were obtained when the biosensor strains were challenged with preservatives, for all except lpp-lux E. coli. The bioluminescence expressed by the lpp-lux biosensor was significantly lower during long-term stationary phase than it was for any of the other biosensors and was also significantly lower than for any of the other biosensors in the presence of preservatives. Since the plasmid copy number and viable counts for lpp-lux did not change under these conditions, it suggests that perhaps lpp-lux was down regulated under stress conditions. There were no statistically significant differences between the results of the bioluminescence assays and the results of the viable count and ATP chemiluminescence assay. Virtual foot printing (using Regulon DB database) demonstrated that two crp binding sites overlapping the -10 regions are located on the negative strand of the lysS promoter sequences and that one crp binding site is located in lpp. The biosensor strains ldc-lux exhibited levels of bioluminescence per cell significantly lower than spc in the presence of preservatives whilst there was a significant increase in bioluminescence per cell by tatA-lux under alkaline conditions (pH 8.9) during long-term stationary phase. Amongst the five biosensor strains tested in the current work, it was determined that the spc-lux strain would be the most attractive candidate for further work, since the bioluminescence expressed per cell was significantly greater, by 10-1000 times, than that expressed by the other four promoters when challenged with the preservatives tested with excellent significant correlations between bioluminescence expression and viable counts in the PET assays with the various preservatives in this study (R2: 8.79-1.00). The bioluminescent biosensor strains showed no statistical differences from the control strains (wildtype E.coli ATCC 8739 and E.coli carrying a promoterless [pBR322.lux] for adneylate energy charge (AEC), plasmid copy number (PCN) bioluminescence or viable counts over 28 days. The emission of bioluminescence by the four bioreporter strains across 28 days is reflected by the stability of PCN with correlations of 0.78-0.90, except for lpp-lux with R2: 0.59. The following promoter elements were found likely to assist greater expression of bioluminescence: an A+T level of approximately 50% between the -40 and -60 regions (the UP element); a G+C level of approximately 50% within the -10 and +1 regions; the extended -10 region and -10 region of consensus sequence RpoD (σ70/D).

Human immunodeficiency virus (HIV) is a retrovirus that can lead to acquired immunodeficiency syndrome (AIDS), a condition in humans in which the immune system begins to fall, leading to life-threatening opportunistic infections.• As of January 2006, the Joint United Nations Programme on HIV/AIDS (UNAIDS) and the World Health Organization (WHO) estimate that AIDS has killed more than 25 million people since it was first identified on December 1, 1981, making it one of the most destructive pandemics in recorded history. Pheroid technology is a patented delivery system that consists of both plant and essential fatty acids. It is often confused with lipid-based delivery system. Although there are some similarities, Pheroid™ technology owns its advantages in terms of absorption and/or efficacy of pharmacologically active compounds and other useful molecules. The Pheroid structure can be manipulated in terms of morphology, structure, size and function. The effectiveness of Pheroid technology has been illustrated by several national and international clinical trials with products based on this technology. Pro-Pheroid production is similar to that of Pheroid production, except that no aqueous phase is introduced; instead, the active compounds are dissolved in the oil phase. This study was conducted to determine the stability of both nevirapine and butyiparaben in the pro-Pheroid delivery system as well as the preservative efficacy of butylparaben. High performance liquid chromatography (HPLC) was used to determine the stability of nevirapine and butylparaben in pro-Pheroid. The formulation was stored under controlled conditions, i.e. 5°C, 25°C + 60% RH, 30°C + 65% RH and 40°C + 75% RH, for three months. The accelerated stability study was performed according to ICH guidelines. The preservative efficacy study was done by the EnviroCare Laboratory according to international specifications. The various studies conducted on nevirapine in pro-Pheroid to determine the stability showed that nevirapine could successfully be formulated in the pro-Pheroid system and that butylparaben is an effective preservative in the formulation. It is finally concluded that before nevirapine in pro-Pheroid is further formulated into viable products, the following issues will have to be addressed: • The pro-Pheroid manufacturing process should be assessed and validated to ensure batch to batch uniformity. • In order to establish a sound analytical method, stability of the pro-Pheroid system (without the addition of pharmaceutical actives) should be evaluated over a period of at least six months. • The specific UV detection wavelengths of both nevirapine and butylparaben should be assessed and validated to get the optimum wavelength for the HPLC assay analysis to ensure the integrity of the results obtained. The physical properties (color, smell and taste) of the pro-Pheroid system need to be addressed to make the product acceptable to the consumer.
Thesis (M.Sc. (Pharmaceutics))--North-West University, Potchefstroom Campus, 2010.

Whole cell biosensors have been extensively used for monitoring toxicity and contamination of compounds in environmental biology and microbial ecology. However, their application in the pharmaceutical and cosmetics industries for preservative efficacy testing (PET) has been limited. According to several pharmacopoeias, preservatives should be tested for microbial activity using traditional viable count techniques; the use of whole cell microbial biosensors potentially provides an alternative, fast, and efficient method. The aim of the study was to construct and develop whole cell microbial biosensors with Pseudomonas aeruginosa ATCC 9027. Constitutive promoters: PlysS, Pspc, Ptat, Plpp and PldcC and the lux-cassette were inserted into plasmid pME4510 and transformed into P. aeruginosa ATCC 9027 cells to produce bioluminescent strains. Plasmids were found to be maintained stably (~50 copies per cell) throughout the growth and death cycle. The novel bioluminescent strains were validated in accordance with the pharmacopoeia using bioluminescence detection and quantification followed by comparison with the traditional plate counting method. The bioluminescent method was found to be accurate, precise and equivalent at a range of 103 – 107 CFU/mL, as compared with plate counting. Recovery of bacterial cells was quantified using bioluminescence; this method proved to be accurate with percentage recoveries between 70-130% for all bioluminescent strains. The method was also more precise (relative standard deviation less than 15%) than the traditional plate counting method or the ATP bioluminescent method. Therefore, the bioluminescent constructs passed/exceeded pharmacopoeial specified criteria for range, limit of detection, accuracy, precision and equivalence. Physiology of the validated bioluminescent strains was studied by assessing the growth and death patterns using constitutive gene expression linked with bacterial replication. Promoter strengths were evaluated at various stages of the growth and death pattern and related to promoter sequences. PlysS, Ptat and Plpp were relatively strong promoters whilst PldcC and Pspc were relatively weak promoters. Relative promoter strength decreased in the order of Plpp>Ptat>PlysS>PldcC>Pspc during the exponential phase whilst Ptat was stronger than Plpp during the stationary phase of growth. Plpp had its highest level of expression during the exponential phase, while Ptat had relatively stable lux expression during the stationary phase. Correlations between relative bioluminescence and CFU at 24 hours were greater than 0.9 indicating a strong relationship for all bioluminescent strains. Reduction in correlation coefficients to approximately 0.6 between relative bioluminescence and CFU and between relative fluorescence and CFU beyond 24 hours indicated that a certain proportion of cells were viable but non-culturable. Tat-pME-lux showed steady bioluminescence compared to CFU count (R>0.9) throughout 28 days of growth. Equivalence analysis showed no significant difference between the bioluminescence and plate count method throughout 28 days of growth for all five bioluminescent strains. Applicability of these novel bioluminescent strains was evaluated for preservative efficacy tests (PET) using bacterial replication and bioluminescence as a measure of constitutive gene expression. PET using benzalkonium chloride and benzyl alcohol showed no significant difference between the bioluminescent method and the plate count method. Good correlations between bioluminescence, CFU count and fluorescence were obtained for benzalkonium chloride (BKC) concentrations (R>0.9) between 0.0003% and 0.0025% against strains lysR25, lppR4 and tatH5. Similarly, good correlations (R>0.9) between the three parameters were obtained for benzyl alcohol (BA) concentrations between 0.125% and 2% against strains lysR25, lppR4 and tatH5. The bioluminescent method and traditional plate counting method were equivalent for concentrations of BKC (0.0003 - 0.02%) and BA (0.25 - 2%) during preservative efficacy tests. These bioluminescent constructs therefore are good candidates for selection for preservative efficacy testing. The bioluminescent method and traditional plate counting method were also found to be equivalent for construct tatH5 at a concentration of 0.125% BA. PET testing with BKC and BA showed that tatH5-pMElux (R>0.9) had consistently high correlation coefficients between CFU and relative bioluminescence. Together with the results from growth and death kinetics, where tatH5 showed the greatest constitutive expression, it can be concluded that P. aeruginosa ATCC 9027 tatH5-pMElux is the best construct for testing various antimicrobial agents. This study has shown that according to the pharmacopoeial requirements, the bioluminescent method is more accurate, precise and equivalent to the traditional plate counting method and therefore can be utilised instead of the traditional plate counting method for the purpose of preservative efficacy testing.

The work described in this thesis was undertaken to determine the nature and identity of soluble carbohydrate and nitrogenous components which migrate and accumulate at evaporative surfaces of dried wood and the influence these nutrients have on wood decay and preservative performance. Specific soluble carbohydrates and amino acids were shown to redistribute and accumulate at surface regions of wood during drying. Analysis of dried wood showed that soluble carbohydrates constituted 2-5% of the dry mass of wood at surface regions, and that soluble nitrogenous components constituted < 0.5% in the same areas. The soluble sugars which redistributed and accumulated at surface regions during drying were mainly reducing in nature in the softwoods. Glucose and fructose were the predominant sugars in these woods. In lime, sucrose was the predominant sugar. Soluble amino acids contributed to a significant proportion of the nitrogen content at surface regions of softwoods. In pine and spruce soluble amino acids constituted 30% and 40% of the total nitrogen content, but in lime, concentrations of soluble amino acids constituted only 6% of the total nitrogren content. The major amino acids observed in pine, spruce and lime were aspartic acid, glutamine and arginine. Soil burial studies undertaken highlighted the problems encountered when trying to mimic natural wood of high nutrient status. Test blocks impregnated with soluble sugars and amino acids displayed loss of these added nutrients on emplacement in soil, and the effect of added substrates could not be evaluated individually. The results of soil burial studies using CCA treated wood which was also impregnated with amino acids, showed that the latter influenced wood decay and preservative stability in lime. Weight losses in preserved lime were shown to correlate with increasing arginine and glutamine concentrations. A substantial copper loss was recorded in hardwoods and softwoods treated at sub-toxic levels with CCA and also treated with glutamine. Soluble sugars incorporated into preserved wood did not influence wood decay or preservative efficacy.

Objectives: Tooth extraction initiates a cascade of events that often leads to local anatomic changes in the alveolar ridge. Ridge preservation is a surgical approach aimed at minimizing hard and soft tissue volume loss. There have been contradicting reports on the efficacy of socket grafting for alveolar ridge preservation. Interestingly, there is a paucity of adequately powered randomized controlled clinical trials. The purpose of this study was to assess the effect of the application of a socket grafting technique on alveolar ridge dimensional changes following tooth extraction. Methods: Healthy patients requiring the extraction of one single-rooted tooth on either arch, from second premolar to second premolar, excluding mandibular incisors, and who met the eligibility criteria were recruited. Patients were then randomly assigned to either the control group, consisting of tooth extraction alone, or the experimental group, which consisted of extraction and simultaneous ridge preservation using an allograft bone material to fill the socket and a dense polytetrafluoroethylene membrane (dPTFE) to seal it. Cone beam computed tomography (CBCT) was obtained immediately prior to extraction (baseline) and at 14 weeks. Linear measurements with the use of a tooth-supported stent were obtained immediately after extraction (baseline) and at 14 weeks. Linear and volumetric measurements were made using data obtained from the CBCTs. Masked, calibrated examiners performed all radiographic measurements. Measurements obtained included buccal keratinized tissue width, buccal and lingual plate height and width, alveolar ridge horizontal width (CBCT); and alveolar ridge volume changes. Digital planning of dental implants was performed in the ideal restorative location and need for additional grafting was virtually determined. The primary outcome of interest was volumetric reduction of the alveolar ridge at 14 weeks. Linear mixed model statistical analyses were used to compare the mean change in the measurements between the grafted and control groups. Results: A total of 59 subjects were recruited, of which 53 patients (27 control and 26 experimental) completed the study. No statistically significant difference was found between the two groups at baseline for any of the parameters analyzed. At the 14 week follow-up appointment there was an average loss in height of the buccal plate of 1.17 mm and 0.61 mm for the control (CG) and experimental (ARP) groups, respectively, showing statistical significance (p=0.012). The lingual plate height was reduced 0.7 mm in CG and 0.47 mm in ARP with no statistical significance (0.075). A linear loss in the buccal-lingual dimension of the alveolar ridge was noted radiographically in both groups, 1.68mm in CG and 1.07mm in ARP, which demonstrated a statistical significant difference between them (p=0.023). Volumetric analysis demonstrated a mean volume loss of 15.83% in the CG showing statistical significance from the 8.36% loss shown in the ARP group. This difference demonstrates a clinical significance when virtual planning of implant placement in the ideal restorative location revealed the need for additional grafting at 13/27 or 48% of CG and 3/26 or 11% of ARP sites. Additionally, a very robust, statistically significant correlation was noted between buccal bone plate width and reduction of alveolar bone volume after 14 weeks of healing (p< 0.0001). A multivariate regression analysis revealed that within the control group a buccal plate <1mm lead to >10% volumetric reduction, while the same reduction in the graft group was only seen when the buccal plate was less that 0.6mm. Conclusions: In this study, a novel volumetric analysis of alveolar ridge reduction after tooth extraction was performed, which demonstrated that socket grafting for alveolar ridge preservation does provide a therapeutic benefit. This finding was associated to a decreased probability of requiring additional grafting at the implant site. The thickness of the buccal plate at the time of extraction appears to be a valuable factor to predict the amount of resorption that will take place, meaning that more resorption should be expected, as the buccal plate gets progressively thinner.

Background: The shortage of kidneys for renal transplantation has prompted renewed interest in non-heart-beating donors (NHBD). While this may increase the number of transplants, it also increases the primary non-function (PNF) rate. This is caused by excessive warm ischaemic injury in some NHBD, and has hindered their more widespread use. A reliable pre-transplant test of organ viability, and a preservation method minimising additional iscahemic damage, would allow the PNF rate to be reduced. The aims of this thesis were to explore the potential of warm ex-vivo perfusion as a preservation method and a means of diagnosing viability pre-transplantation. Methods: Warm ex-vivo perfusion of ischaemically injured porcine kidneys with an oxygenated emulsion of a perflourochemical in tissue culture fluid was used to measure ex-vivo function and preserve kidneys. A cadaveric model was used to assess the relationship of ex-vivo function and warm ischaemic time. An autotransplant model was used to determine the relationship of ex-vivo to post-transplant function, and to compare the efficacy of preservation by warm perfusion with conventional hypothermic techniques of static storage and pulsatile perfusion. Post-transplant outcome measures were survival, renal function and histology. Results: WIT correlated well with ex-vivo function. Ex-vivo function correlated with post-transplant function in terms of survival (and therefore the immediate life supporting function of the kidneys), but not to the extent that it could be used to predict viability better than knowing the WIT alone. The efficacy of warm perfusion was indistinguishable from hypothermic static storage. However hypothermic pulsatile perfusion was slightly superior to both other techniques. Conclusions: Warm perfusion as used in this thesis was broadly equivalent in efficacy to conventional hypothermic organ preservation techniques. Although ex-vivo function correlated with post-transplant function, the correlation was not tight enough to support a diagnostic role for ex-vivo function in viability determination.

Hazard Analysis Critical Control Point (HACCP) regulations for juice manufacture require the application of a process that will result in a 5-log reduction (99.999%) of the pertinent pathogen in the juice being processed. The use of ultraviolet (UV) light, as an alternative to traditional thermal processing, has been adopted by some juice processors as a means of meeting the HACCP 5-log performance standard. However, little research had been performed to determine the effect of UV when used in combination with antimicrobial agents that are commonly added to juice products. Therefore, the objectives of this work were (1) to determine if chemical preservatives and ultraviolet light have a combined effect on the reduction of Escherichia coli in apple cider, and (2) to determine the influence of adding chemical preservatives at different points in the processing of juice (i.e., either prior to or after ultraviolet light processing) on the reduction of Escherichia coli in apple cider. In this study, refrigerated (4°C) pasteurized apple cider that contained no added preservatives was inoculated with E. coli ATCC 25922, a surrogate strain for E. coli O157:H7, and exposed to UV (peak output: 254 nm). The following chemical preservatives were added to apple cider either prior to or after UV exposure: dimethyl dicarbonate (75 and 150 ppm), hydrogen peroxide (75 and 150 ppm), potassium sorbate (1000 and 2000 ppm), and sodium benzoate (1000 and 2000 ppm). Following UV exposure and chemical preservative application, inoculated juices were stored at 4°C for 72 hours. Samples were collected prior to and immediately after UV exposure and at 24, 48, and 72 hours of storage. At each sampling point, juice portions (0.1 ml) were serially diluted in peptone diluent (0.1%) and surface plated onto Tryptic Soy Agar (TSA). Counts of the bacterial colonies were made 48 hours after incubating plates at 35°C. Overall, reductions of E. coli were greater in cider treated with preservatives after UV processing than when preservatives were added prior to UV processing (P < 0.05). Furthermore, dimethyl dicarbonate and hydrogen peroxide were more effective than potassium sorbate and sodium benzoate in reducing E. coli populations in conjunction with UV (P < 0.05). When added prior to UV exposure, potassium sorbate was the least effective, allowing for the greatest survival (P < 0.05). This study describes the use of UV in conjunction with hydrogen peroxide and dimethyl dicarbonate as an effective method for producing a 5-log or greater reduction of E. coli O157:H7 in apple cider.
Master of Science

The effect of pressure on the log reduction of six strains of E. coli O157:H7 and five serovars of Salmonella enterica were investigated in tryptic soy broth, sterile distilled water and commercially sterile orange and apple juice. Samples were subjected to high hydrostatic pressure (HHP) at 300 and 550 MPa for 2 minutes at 6°C, and then held for 24 hours at 4°C following treatment. E. coli O157:H7 strain E009 was the most pressure resistant, having a decrease of only 0.77 log10 CFU/ml directly after pressurization in TSB. S. Agona was the most pressure resistant Salmonella serovar tested with a decrease of 3.79 log¬10 CFU/ml in TSB at 550 MPa. The two most pressure resistant cultures were then used in a subsequent study using HHP in conjunction with antimicrobials (dimethyl dicarbonate [DMDC] at 62.5 and 125 ppm, hydrogen peroxide at 150 and 300 ppm, cinnamic acid, potassium salt at 125 and 250 ppm, potassium sorbate [KS] at 500 and 1000 ppm and sodium benzoate [NaB] at 500 and 1000). For both E. coli O157:H7 and Salmonella, the most effective antimicrobial was DMDC having a 5.79 and 5.96 log10 CFU/ml decrease directly following pressurization, respectively. Other treatments that were significantly different from the samples treated with no antimicrobial were hydrogen peroxide, and NaB at 500 ppm for E. coli O157:H7 and a treatment of NaB at 1000 ppm for S. Agona. After 24 hours at 4°C, S. Agona samples with added antimicrobials had a close to or above 5-log10 CFU/ml reduction. DMDC should be further investigated as an antimicrobial agent that can work in conjunction with HHP.
Master of Science

碩士
國立高雄海洋科技大學
水產食品科學研究所
101
Histamine poisoning is usually associated with fishes such as tuna or sailfish. Because those fishes contain high levels of free histidine which can be converted by a decarboxylase produced by bacteria. Therefore, inhibiting the growth or enzymatic activities of histamine producing bacteria (HPB) is a critical procedure to prevent histamine poisoning. Japanese horseradish (wasabi) is a popular spice in Taiwan and its major volatile compound, allyl isothiocyanate (AIT), has been reported to possess a high antibacterial activity. Currently, vacuum-packing is the most common method to extend the shelf life of fishery products. However, a single preservation method such as applying AIT or vacuum packing is insufficient. The antibacterial and preservative efficacy of the combination of vacuum packaging and gaseous AIT in sailfish was evaluated. In the study, sailfish samples were inoculated with histamine producing bacteria, Raoultella ornithinolytica, then treated with different concentrations of gaseous AIT (329.08, 459.46 or 516.70 mg / L air) at 4°C for 24 hr. The sailfish samples were vacuum or non-vacuum packed, then stored at 15°C, 25°C or 4-25-4°C. The samples without vacuum package no AIT treatment were served as the control group. Levels of total aerobic counts, histamine-producing bacteria, total volatile basic nitrogen (TVBN) and biogenic amines were analyzed daily. Significantly lower counts of total aerobic and histamine producing bacteria was found in the AIT treated samples than the samples without AIT treatment (p<0.05) within the group without vacuum packaging. However, bacterial counts were not significantly reduced within the group with vacuum packaging. In addition, similar results were obtained for TVBN. TVBN values reduced correlated with the increasing AIT concentrations in the unvacuum packed group but not in the vacuum packed group. In the unvacuum packed group, histamine amount reduced significantly with AIT treatment (p<0.05) and the reduction is correlated with AIT concentrations. In the vacuum packed group, AIT treatment also showed an inhibitory effect against histamine production. Comparing the control, the vacuum packaged samples showed significantly lower histamine production than unvacuum packaged samples (p<0.05).

The efficacy of biodiesel as a co-solvent for copper naphthenate wood preservative treating solutions was evaluated using two fungal decay methodologies (AWPA E10-09, British Standard Method EN113). Four fungal species (Gloeophyllum trabeum, Trametes versicolor, Poria xantha, Postia placenta) and three wood species (Douglas fir, Southern yellow pine, Western red cedar) with six replicates were utilized in both studies. Two levels of biodiesel: diesel (30:70 and 50:50) were compared to diesel-only solvent systems for copper naphthenate treating systems and treated to AWPA recommended retentions. No differences in decay efficacy between the biodiesel blends and diesel-only treatment in either the AWPA or the EN113 decay studies were detected for either standard method. Copper distribution was evaluated using SEM-EDX and no differences were noted with either solvent system. It was determined that the presence of biodiesel did not have a negative impact upon the efficacy of copper naphthenate as a wood preservative.

碩士
國立臺灣海洋大學
水產養殖學系
102
In this study, intends to investigate the effect of preservation and efficacy of multiemulsified oral inactivated vaccine against grouper iridovirus. Firstly experimental inactivated Grouper iridovirus (GIV) vaccines were prepared, after safety testing, stored at 4℃ and 25℃ room temperature for 8 months and 12 months, multie-mulsified vaccine are not occurring disintegrations. Further compared the efficacy with the new preparation vaccine protective, we had mix these five vaccines into fish feed and fed once every other week. Challenge with GIV at day 10 and day 20 post vaccination. The relative survival rate of the new preparation vaccine are 60% and 50%, of the vaccine preserved at 4 ℃ and 25 ℃ for 8 month are around 40 %, and of 4 ℃ and 25 ℃ for 12 month are decreased to 20%. Analyzed the anti GIV antibody titre in serum from each groups at day 1, 4, 7 post vaccination, the results which had compared with non-vaccinated control group indicated that antibody had stimulated in all the vaccinated fish. The antibody titre had slightly decreased at day 10, 14 post vaccination were observed in all the groups, especially the group fed with vaccine had preserved at 4 ℃ and 25 ℃ for 12 month. Finally, we analyzed the gene expression of MHC-I, TNF-α, IgM, Mx and INF-γ in spleen and head kidney. The results suggest that vaccinated groups were much higher than non-vaccinated groups, such as INF-γ in spleen. Mx gene expression has significant difference between new prepared vaccine and other preserved vaccines. All the results had suggested that the inactivated vaccine could be well preserved using multie- mulsified preperation, and still could stimulate immune response after storing at 4℃ for 8 month.

碩士
國立臺灣大學
臨床牙醫學研究所
105
Background: Alveolar ridge remodeling is observed after tooth extraction. It would become the problem if future rehabilitation involved implant placement. Therefore , ridge preservation technique was introduced to solve this kind of problem. Although there are many available products of β-TCP(β-tricalcium phosphate)/Collagen composites for clinical usage, such as for periodontal regeneration and repair of bony defect. There is still lack of ideal products fulfilled the requirement of easily handling and predicable clinical outcome. Some popular product such as Bio-Oss® Collagen may retained too long during its degradation. It may degrade relatively slowly, more residual bone filler after bone modeling. β -TCP also has substantial physical strength; it provides a three-dimensional scaffold for bone regeneration against the pressure of tissue shrinkage . Moreover, β-TCP has the potential to function as a reservoir of calcium and phosphate ions for the local tissue during the degradation process, which possibly results in stimulation of osteoblastic function and promotion of bone formation. Above as though better than traditional β-TCP/Collagen composites. In this project, we will perform animal test to identify its clinical efficacy. Different β-TCP/Collagen composites will be tested in their physical property. Afterward proceeding for further animal experiment in observation the dimensional change of the extraction socket. Six dogs will be used in this project, symmetric extraction sockets will be created and covered with commercial or new developed β-TCP/Collagen composites in this project at different time points to identify its efficacy in ridge augmentation. We hope the β-TCP/Collagen composites will be developed and its superiority in guide bone regeneration will also been identified following this project proceeding. Materials and Methods: The mandibular third premolars and fourth premolars of six dogs were extracted bilaterally, and buccal dehiscence defects were prepared bilaterally on the buccal side of the mandibular third and fourth premolars. Sixteen defects were assigned to four treatment groups: T1 group (β-TCP/10%Collagen composites );T2 group (β-TCP/20%Collagen composites ) ; positive control (Bio-Oss® Collagen); negative control (no grafting material ). Fluorescence bone labeling was administrated subcutaneously three weeks, five weeks and seven weeks post-operatively. The animals were sacrificed 8 weeks after surgery. For histomorphometric analysis, new bone area were measured. New bone volume was measured using microcomputed tomography(micro-CT). Fluorescence microscopic observation was performed to figure out the possible sequence of new bone formation in each group. Result: No adverse effect was found in all four groups after ridge preservation surgery. For new bone area (%) of Histomorphometry, the four groups was 25.02±19.50% for the T1 group, 38.18±6.85% for the T2 group , 32.37±10.74% for the positive control group, and 24.97±8.91% for the negative control group. The differences between each group were not statistically significant. For new bone volume (%)analyzed from micro-CT, the result of the T1 group come close to the positive control group (respectively 71.68±12.63 % and 82.40±14.14 %), which is higher than T2 group (62.00±14.39 %) and the negative control group (31.18±12.57 %).Nevertheless, there is statistical significant between positive control、T1 group and negative control group. Under fluorescence microscopy, the pattern of new bone formation usually starts from the border of the defect toward the central part of the defect. However, the timing and the specific area of new bone formation are different in positive control、T1、T2 and negative control groups. It seems that the test group may delay the time of bone formation (compare with negative control group) ,but more bone gain may find after 5 weeks in the test group. For new bone area (%)of fluorescence microscopy calculated separately in the four groups was 13.88±3.15 % for the T1 group, 12.42±8.37 % for the T2 group , 15.24±8.85 % for the positive control group, and 6.33±3.73% for the negative control group. But the differences between each group were not statistically significant. Conclusion: The β-TCP/Collagen composites seems to have the potential to preserve ridge volume. However, due to limited sample size in this study, there is less possibility that the result between groups achieving statistical significance. More research are needed on investigating the potential of this new β-TCP/Collagen composites for ridge preservation. In addition, the fluorescence bone labeling techniques not only supporting the results of histology and micro-CT , but also assisting us to understand the mechanism of new bone formation in different treatment

Dissertação de Mestrado apresentada ao ISPA - Instituto Universitário
Tratou-se de um estudo no qual participaram 120 jovens com idades compreendidas entra os 18 e os 30 anos, cujo principal objectivo foi investigar a relação existente entre ambos os géneros e a auto-eficácia ao preservativo masculino nos relacionamentos sexuais. Foram utilizados dois grupos, um feminino composto por 60 raparigas, outro masculino composto por 60 rapazes. Para medir a auto-eficácia foi utilizada uma escala já existente, adaptada para a língua portuguesa, nomeadamente a Escala de Auto-Eficácia Face ao Preservativo de 20 itens de Marín et al. (1998). Em termos gerais pretendeu-se observar se existiam diferenças significativas entre os dois grupos no que respeita às duas variáveis principais auto-eficácia e género e que outras variáveis poderiam estar a influênciar os resultados obtidos. O estudo decorreu no meio estudantil, nomeadamente com estudantes universitários.
This was a study in which we had 120 young person’s participating, with an average age between 18 and 30 years old. The main objective of this study was to investigate the existing relation between both genders and self-efficacy in male condom use in sexual relationships. They were used two groups, one feminine composed by 60 young women, and the other, a male group composed by 60 young men. To measure self-efficacy it was used an existing scale, adapted to Portuguese language, the 20-item Self-Efficacy to Use Condom Scale of Marín et al. (1998). In general terms the objective of this study was to observe the significant differences between the principal variables, self-efficacy to use condom and gender, and to observe what other variables influence results. The study was carried out on the student ambience, with university students.

A dissertation submitted to the Faculty of Health Sciences, University of Witwatersrand, in fulfilment of the requirements for the degree of Master of Science (Medicine) in the branch of Anatomical Pathology Johannesburg, 2012
Objective To determine the effectiveness of the cell block technique for immunocytochemical diagnosis by comparing cytomorphologic preservation and immunocytochemistry (ICC) stains in paired cell block and conventional fine needle aspiration (FNA) samples. Study Design This was a prospective study. Material for both conventional smears and cell blocks were collected simultaneously during fine needle aspiration of 50 lesions comprising lymph node, lung and liver masses. Grading of cellularity, morphological preservation, architectural preservation, immunocytochemical staining intensity and presence of background staining were compared on paired FNA smears and cell block samples derived from the same case. Each arm of the paired analysis was performed blindly without knowledge of the grading outcome of the other. The Kappa statistic (Κ) was used to measure inter-rater agreement. Results The fifty samples evaluated included FNAs from the lung, 24/50 (48%); liver, 23/50 (46%) and lymph node, 3/50 (6%). The immunocytochemistry stains consisted of 44/50 (88%) CK7, 44/50 (88%) CK20, 18/50 (36%) TTF1, 10/50 (20%) synaptophysin, 10/50 (20%) Hepar-1 and 7/50 (14%) AE1/3. There was no overall agreement in preservation of cytomorphological detail and ICC staining between the two methods. The Papanicolaou stained conventional FNA smears fared better then cell block for the vi evaluation of nuclear and cytomorphologic characteristics; cells in the cell block were poorly preserved in many cases. The ICC stains worked better on the cell block samples due to lack of background and aberrant staining. Conclusion Conventional FNA smears and cell blocks complement each other. Our results indicate that it would be optimal to use both modalities in the diagnostic work-up of mass lesions amenable to FNA diagnosis; the former to assess morphology, and the latter for optimal immunocytochemistry results. In resource constrained settings, the cost implications of performing both conventional and blocked smears on all FNA material warrants further evaluation.

Indiana University-Purdue University Indianapolis (IUPUI)
Colon rectal cancer (CRC) is one of the most common cancers worldwide. It is characterized by the successive accumulation of mutations in genes controlling epithelial cell growth and differentiation leading to genomic in-stability. This results in the activation of proto-oncogene(K-ras), loss of tumor suppressor gene activity and ab-normality in DNA repair genes. Targeted therapy is a new generation of cancer treatment in which drugs attack targets which are specific for the cancer cell and are critical for its survival or for its malignant behavior. Survival of metastatic CRC patients has approximately doubled due to the development of new combinations of stan-dard chemotherapy, and the innovative targeted therapies, such as monoclonal antibodies against epidermal growth factor receptor (EGFR) or monoclonal antibodies against vascular endothelial growth factor (VEGFR).The study is to exhibit the need for right drug-right target and provides a proof of principle for potent efficacy of molecular targeted therapy for CRC. We have performed the weighted gene co-expression network analysis for three different patient cohort treated with different targeted therapy drugs. The results demonstrates the variation across different treatment regime in context of transcription factor networks. New significant tran-scription factors have been identified as potential biomarker for CRC cancer including EP300, STAT6, ATF3, ELK1, HNF4A, JUN, TAF1, IRF1, TP53, ELF1 and YY1. The results provides guidance for future omic study on CRC and additional validation work for potent biomarker for CRC.

Research Doctorate - Doctor of Philosophy (PhD)
In Chapter 1, I describe social competence, task competence and self-protection in an organisational context. In Chapter 2, I review key self theories and relate them to the self-competence construct. In Chapter 3, I review the research on self-competence to show that there is a need for a construct of social competence and self-protection. I discuss the limitations of three self-competence theories: Bandura’s (1977) self-efficacy theory, Williams and Lillibridge’s (1992) self-competence theory and Tafarodi & Swann’s (1995) self-competence/self-liking theory. In Chapter 4, I present my selfcompetence model. I raise the research questions and specify my hypotheses. In Chapter 5, I describe the construction of Social and Task Competence Scale. I present evidence of the reliability and factor structure of the Social and Task Competence Scale. I concluded that scale revisions were needed. In Chapter 6, I present evidence of the reliability, factor structure and predictive validity of the revised Social and Task Competence Scale and Self-Protection Scale. I describe the results of an experiment that investigated the interaction of task setting, social competence, task competence and selfprotection. I concluded that the measures predicted performance. In Chapter 7, I investigate the factor structure and reliability of the revised Social and Task Competence Scale and revised Self-Protection Scale. I provide evidence of the convergent and discriminant validity of these measures with reliable measures of self-competence, selfesteem, self-monitoring, personality and social desirability. In Chapter 8, I investigate the factor structure and reliability of the Social and Task Competence Scale and Self-Protection Scale after final revisions and show that these measures are acceptable for use in scientific research. I present evidence of their convergent validity with a valid andreliable measure of emotional intelligence, and describe experimental results that supported the hypothesised relationships between perceived task difficulty, social competence, task competence and self-protection and task performance. In Chapter 9, I discuss the implications of my research for self-competence theory, self-regulation and self-esteem and the prediction of social and task performance in organisations.

Research Doctorate - Doctor of Philosophy (PhD)
In Chapter 1, I describe social competence, task competence and self-protection in an organisational context. In Chapter 2, I review key self theories and relate them to the self-competence construct. In Chapter 3, I review the research on self-competence to show that there is a need for a construct of social competence and self-protection. I discuss the limitations of three self-competence theories: Bandura’s (1977) self-efficacy theory, Williams and Lillibridge’s (1992) self-competence theory and Tafarodi & Swann’s (1995) self-competence/self-liking theory. In Chapter 4, I present my selfcompetence model. I raise the research questions and specify my hypotheses. In Chapter 5, I describe the construction of Social and Task Competence Scale. I present evidence of the reliability and factor structure of the Social and Task Competence Scale. I concluded that scale revisions were needed. In Chapter 6, I present evidence of the reliability, factor structure and predictive validity of the revised Social and Task Competence Scale and Self-Protection Scale. I describe the results of an experiment that investigated the interaction of task setting, social competence, task competence and selfprotection. I concluded that the measures predicted performance. In Chapter 7, I investigate the factor structure and reliability of the revised Social and Task Competence Scale and revised Self-Protection Scale. I provide evidence of the convergent and discriminant validity of these measures with reliable measures of self-competence, selfesteem, self-monitoring, personality and social desirability. In Chapter 8, I investigate the factor structure and reliability of the Social and Task Competence Scale and Self-Protection Scale after final revisions and show that these measures are acceptable for use in scientific research. I present evidence of their convergent validity with a valid andreliable measure of emotional intelligence, and describe experimental results that supported the hypothesised relationships between perceived task difficulty, social competence, task competence and self-protection and task performance. In Chapter 9, I discuss the implications of my research for self-competence theory, self-regulation and self-esteem and the prediction of social and task performance in organisations.