Programmed cell death (PCD) is an indispensible process in plant and animal systems that serves to eliminate cells and/or tissues and recycle nutrients from these tissues to the rest of the organism. In animals, PCD is referred to as apoptosis and is performed by caspases, a family of aspartate-specific cysteine proteinases that serve to perceive the cell death signal and execute the cell death phenotype. In 2000, Uren et al. discovered a new family of cysteine proteinases in plants called metacaspases – distant arginine/lysine-specific relatives of animal caspases – thought to be involved in plant PCD. The goal of this study was to correlate SlMCA2 expression with PCD in tomato. Polyclonal antibodies were obtained against the Type I metacaspase SlMCA2 in Solanum lycopersicum and used for Western blot analyses. BY-2 cell biolistics and in-situ hybridization were used to investigate where SlMCA2 protein and mRNA accumulate in various tissues. Results produced were replicated a minimum of three times and correlate SlMCA2 to PCD, but not initiation of PCD.

No description available.

Hemoglobins (Hbs) are heme-containing proteins belonging to the globin superfamily that are ubiquitous in most living organisms including prokaryotes and eukaryotes. In addition to the first legHbs found in leguminous plants, there are another three classes of phytoglobins (Pgbs) identified in various plant species including dicots and monocots. The ability of heme groups to bind gaseous ligands such as oxygen, carbon monoxide and nitric oxide (NO) places Pgbs as multifunctional players in various processes during plant growth and development under normal or stress conditions. The objective of this project is to investigate how transcriptional manipulation of ZmPgb1.1 and ZmPgb1.2 influences somatic embryogenesis in maize (Zea mays). Suppression of either of the two genes is sufficient to induce programmed cell death (PCD) through a pathway initiated by accumulation of nitric oxide (NO) and zinc (Zn2+), and mediated by production of reactive oxygen species (ROS). The effect of the death program on the fate of the developing embryos is dependent upon the localization patterns of the two Pgbs. During somatic embryogenesis, ZmPgb1.2 transcripts are restricted to a few cells anchoring the embryos to the subtending embryogenic tissue, while ZmPgb1.1 transcripts extend to several embryonic domains. Suppression of ZmPgb1.2 induces PCD in the anchoring cells allowing the embryos to develop further, while suppression of ZmPgb1.1 results in massive PCD leading to embryo abortion. Cells suppressing the Pgb genes are also depleted of endogenous auxin (indole-3-acetic acid, IAA) localization established by polar auxin transport (PAT), thus suggesting a possible involvement of this plant hormone in the observed processes. Collectively, it appears that the cell specific expression of Pgbs has the capability to determine the developmental fate of embryogenic tissue during maize somatic embryogenesis through their effect on PCD. This novel regulation has implications for development and differentiation in other species.

No description available.

Thesis (Ph. D.)--University of Wyoming, 1982. / Typescript. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 49-55).

Thesis (Ed. D.)--University of Nevada, Reno, 1982. / "May 1982." Includes bibliographical references (leaves 97-104). Online version available on the World Wide Web.

Made available in DSpace on 2014-06-11T19:29:17Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-06-26Bitstream added on 2014-06-13T18:58:59Z : No. of bitstreams: 1 rodello_l_me_botfmvz.pdf: 226950 bytes, checksum: b009f4db5df84afd8c9c3af24e35fe7b (MD5) / Universidade Estadual Paulista (UNESP) / Objetivou-se avaliar e validar o sistema automatizado de refrigeração e congelação de fabricação nacional TK 3000, comparativamente ao sistema de refrigeração e de congelação pelo método geladeira/vapor de nitrogênio líqüido. Foram utilizados dez carneiros da raça Santa Inês sendo colhidos, por meio de vagina artificial, três ejaculados de cada reprodutor. Após as avaliações do ejaculado, o sêmen foi diluído em meio para congelação Glicina-Gema-Leite e envasado em palhetas de 0,25 Ml com 100 x 106 espermatozóides totais/doses, sob temperatura de 32° C. As amostras foram refrigeradas em geladeira e/ou com controle automatizado e posteriormente congeladas em vapor de nitrogênio líqüido e/ou com controle automatizado. Os resultados foram similares (P<0,05) entre a refrigeração em geladeira e a refrigeração com controle automatizados em todos os parâmetros avaliados pós descongelação e ao final do teste de exaustão, o mesmo observou-se entre a refrigeração em geladeira e congelação em vapor de nitrogênio líqüido e refrigeração e congelação com controle automatizado e as combinações refrigeração em geladeira e congelação com controle automatizado e refrigeração com controle automatizado e congelação em vapor de nitrogênio líqüido. Concluí-se que o sistema automatizado TK 3000 é adequado a criopreservação do sêmen ovino, com qualidade equivalente àquela processada no sistema geladeira/vapor de nitrogênio líqüido. / The objectives of the present study were to validate and evaluate a programmed automatic system for freezing and cooling, using a national equipment TK 3000, compared to a method using refrigerator and liquid nitrogen for the same procedures. Ten rams from Santa Ines breed were submitted to semen collection by artificial vagina, with a total of three ejaculates from each sire. After the ejaculate evaluation, semen was diluted in a freezing extender Egg- Glycine - Milk and stored in a 0.25 mL straws with a total of 100 x 10 6 sperm per dose at 32° C. The samples were cooled inside a refrigerator and /or utilizing an automatic programmed system, being then frozen in liquid nitrogen vapor and/ or using automatic control. The results were simmilar (P< 0.05) between samples submitted to cooling using refrigerator and automatic control for all analyzed parameters after thawing and at the end of the resistant test. The same was observed among samples submitted to cooling in refrigerator followed by liquid nitrogen vapor for freezing, samples cooled and frozen both using an automatic control, samples processed by the combinations of cooling in refrigerator followed by freezing with an automatic control, and samples cooled with automatic control followed by freezing in liquid nitrogen vapor. In conclusion, the TK 3000 is an adequate automatic system for ovine semen cryopreservation and it provides a quality which is equivalent to semen processed in refrigerator/ liquid nitrogen vapor system.

Orientador: Sony Dimas Bicudo / Banca: Carlos Antonio de Miranda Bomfim / Banca: André Luis Rios Rodrigues / Resumo: Objetivou-se avaliar e validar o sistema automatizado de refrigeração e congelação de fabricação nacional TK 3000, comparativamente ao sistema de refrigeração e de congelação pelo método geladeira/vapor de nitrogênio líqüido. Foram utilizados dez carneiros da raça Santa Inês sendo colhidos, por meio de vagina artificial, três ejaculados de cada reprodutor. Após as avaliações do ejaculado, o sêmen foi diluído em meio para congelação Glicina-Gema-Leite e envasado em palhetas de 0,25 Ml com 100 x 106 espermatozóides totais/doses, sob temperatura de 32° C. As amostras foram refrigeradas em geladeira e/ou com controle automatizado e posteriormente congeladas em vapor de nitrogênio líqüido e/ou com controle automatizado. Os resultados foram similares (P<0,05) entre a refrigeração em geladeira e a refrigeração com controle automatizados em todos os parâmetros avaliados pós descongelação e ao final do teste de exaustão, o mesmo observou-se entre a refrigeração em geladeira e congelação em vapor de nitrogênio líqüido e refrigeração e congelação com controle automatizado e as combinações refrigeração em geladeira e congelação com controle automatizado e refrigeração com controle automatizado e congelação em vapor de nitrogênio líqüido. Concluí-se que o sistema automatizado TK 3000 é adequado a criopreservação do sêmen ovino, com qualidade equivalente àquela processada no sistema geladeira/vapor de nitrogênio líqüido. / Abstract: The objectives of the present study were to validate and evaluate a programmed automatic system for freezing and cooling, using a national equipment TK 3000, compared to a method using refrigerator and liquid nitrogen for the same procedures. Ten rams from Santa Ines breed were submitted to semen collection by artificial vagina, with a total of three ejaculates from each sire. After the ejaculate evaluation, semen was diluted in a freezing extender Egg- Glycine - Milk and stored in a 0.25 mL straws with a total of 100 x 10 6 sperm per dose at 32° C. The samples were cooled inside a refrigerator and /or utilizing an automatic programmed system, being then frozen in liquid nitrogen vapor and/ or using automatic control. The results were simmilar (P< 0.05) between samples submitted to cooling using refrigerator and automatic control for all analyzed parameters after thawing and at the end of the resistant test. The same was observed among samples submitted to cooling in refrigerator followed by liquid nitrogen vapor for freezing, samples cooled and frozen both using an automatic control, samples processed by the combinations of cooling in refrigerator followed by freezing with an automatic control, and samples cooled with automatic control followed by freezing in liquid nitrogen vapor. In conclusion, the TK 3000 is an adequate automatic system for ovine semen cryopreservation and it provides a quality which is equivalent to semen processed in refrigerator/ liquid nitrogen vapor system. / Mestre

Abstract Failure to maintain an appropriate balance between cell death and proliferation is partly due to derangements in the regulation of apoptosis. In this work, apoptosis and the expression of apoptosis regulating proteins were studied by 3' - end labeling of fragmented apoptotic DNA (TUNEL) and immunohistochemistry in a set of 147 tissue samples consisting of 44 biopsies of normal and dysplastic bronchial epithelium, and 103 non-small cell lung carcinomas. The quantity of apoptotic cells and bodies, apoptotic index (AI%), is defined as a percentage of apoptotic cells in the entire tumor cell population. Changes in the apoptotic activity were already seen in the metaplasia-dysplasia-carcinoma sequence of the lung, where the AI% increased gradually until moderate epithelial dysplasia but started to decrease after that. Thus, the AI% for invasive NSCLC (1.20 for squamous cell carcinoma and 1.24 for adenocarcinoma) was slightly lower than in premalignant bronchial epithelium (mean 1.50), but clearly higher than in normal tissue (0.20 for normal bronchial epithelium and 0.24 for lung interstitial cells). 53% of SQCCs and 50% of ACs showed p53 positive nuclei indicative of mutated p53 protein. The immunostaining of bcl-2, bax and mcl-1 revealed diffuse, cytoplasmic staining and was present in most tissues studied. No statistically significant associations between the extent of apoptosis and the expression of p53, bcl-2, bax, or mcl-1 could be found, although . The immunostaining for caspases 3, 6 and 8 was restricted to the tumor areas, reflecting increased apoptotic activity in them. The AI% was significantly higher in NSCLCs in which the single-cell staining pattern for caspase-8 was dominant (P = 0.017), whereas the expression of caspases 3 and 6 had no association with apoptosis. The number of apoptotic cells was significantly higher in NSCLC tumors with a high number of CD3+ and CD8+ T-lymphocytes (P = 0.01) and B-cells (P = 0.05). By multivariate analysis, enhanced apoptosis in NSCLC showed a 1.9-fold risk (95% CI 1.04–3.60; P = 0.04) and p53 positivity a 2.3-fold risk (95% CI 1.30–4.10; P = 0.005) for a shortened survival. Both factors appeared as independent prognostic variables. Apoptosis is clearly enhanced in premalignant and malignant lung tissue in comparison with normal tissue. Furthermore, the expression of the apoptosis-regulating genes is different in tumor tissue from that in normal tissue, and some of the changes in their expression can be seen even in the premalignant lesions of the bronchial epithelium. The expression of caspases seen only in tumor tissue implies the activation of the apoptotic mechanisms and, thus, the lowered treshold of tumor cells to undergo apoptosis. Even in the advanced stages of the disease, the immune defense is effective and the cytotoxic action of activated CD8+ T-cells clearly involves apoptosis. Based on these results it is concluded that alterations in the apoptotic activity and changes in the expression of apoptosis-regulating genes are associated with malignant transformation and growth in lung tissue.

Master of Science / Department of Animal Sciences and Industry / Dale A. Blasi / Four pen studies and one digestibility trial were conducted to evaluate the effects of energy level and intake of corn by-product based diets on newly received growing cattle. In Exp.1 there were four diets where one was offered for ad libitum intake and formulated to supply 0.99 Mcal NEg/kg DM (0.99/100) and the other three treatments were fed at 95, 90, and 85% of the ad libitum treatment and to supply 1.10 (1.10/95), 1.21 (1.21/90), and 1.32 Mcal NEg/kg DM (1.32/85), respectively. ADG was unaffected by treatment (P = 0.32). However, G:F increased linearly with increasing energy and decreasing intake level (P < 0.01). In Exp. 2, a digestibility trial was conducted to study diets from Exp. 1. Ruminal propionate linearly increased with increasing dietary energy and decreasing intake (P < 0.01). Total tract DM digestibility increased linearly with increasing energy and decreasing intake (P < 0.01), whereas passage rate decreased (P < 0.01). Experiment 3 validated results from Exp. 1 feeding the 1.10/95 treatment at 2.40% of BW daily and the 1.32/85 treatment at 2.2% of BW daily and studied a DNA-immunostimulant (Zelnate, Bayer Animal Health, Shawnee Mission, KS). Zelnate had no effect on parameters measured. ADG was not different between energy treatments (P = 0.75), but efficiency was greater for the 1.32/85 treatment (P = 0.03). Experiment 4 was designed to observe effects of the 1.32 Mcal NEg/kg DM diet fed at four intake levels of 1.9, 2.2, 2.5, and 2.8 % of BW daily. ADG increased linearly with increasing intake (P < 0.01), however G:F was not affected (P = 0.98). In Exp. 5 a factorial design was employed to evaluate the effects of two by-products; wet corn gluten feed and wet distiller’s grains plus solubles, and two levels of corn processing; whole corn or dry-rolled corn. Final ADG and G:F were not affected by by-product, corn processing, or their interaction (P > 0.30). Additionally, animals and diets from Exp. 1 were used to study effects on antibody production, acute phase protein response, stress, and immunocompetency of healthy and morbid cattle. Diet had no effect on the parameters measured (P > 0.10). A quadratic response to time (P < 0.01) was detected for haptoglobin, titers for bovine viral diarrhea type 1 (BVD-1), and infectious bovine rhinotracheitis (IBR). Haptoglobin was highest on d 14, and close to baseline levels by d 27. Titer levels for BVD-1 and IBR were higher on d 14, and significantly higher on d 27. Titers for bovine viral diarrhea type 2 (BVD- II) responded linearly (P < 0.05) to time with the highest levels on d 27. Haptoglobin was elevated in morbid animals compared to healthy pen mates (P < 0.05). Titer levels for BVD-I and IBR were higher in healthy animals (P < 0.01). Fecal cortisol was higher on arrival than on d 14 (P < 0.05). In summary, high-energy limit-fed diets based on corn by-products do not affect health and are more efficient than when roughage-based growing diets are fed.