Thesis (Ph.D.)--Boston University / The potential usefulness of the membranous cheek pouch of the hamster as an investigative tool in experimental embryology has been explored* After preliminary studies involving the transplantation of ova, morulas, and various embryonal tissues, the anterior limb buds of embryos in the eighth to eleventh day of development were chosen for intensive study.

Thesis (Ph.D.)--Boston University / PLEASE NOTE: Boston University Libraries did not receive an Authorization To Manage form for this thesis or dissertation. It is therefore not openly accessible, though it may be available by request. If you are the author or principal advisor of this work and would like to request open access for it, please contact us at open-help@bu.edu. Thank you. / Recent experimental work has indicated that the thymus in late fetal and early neonatal life plays a major role in the development of mechanisms of immunity. To date, however, no study of the ultrastructure of the prenatal thymus has been reported in the literature, and histochemical and cytochemical studies of this early period are fragmentary. Therefore, in an effort to extend present knowledge of thymic differentiation and function during its early development, the thymus in the golden hamster (Mesocricetus auratus) was studied by selected histochemical methods and by electron microscopy from 9 1/2 days post coitum through 24 hours post partum. Histochemical techniques for the light microscopy included the periodic acid-Schiff technique with salivary digestion control for glycogen, the methyl green-pyronin technique as an indicator for protein synthesis, the sudan black B technique for determination of total unbound lipid, the Nile blue and oil r ed 0 techniques for deter mination of neutral lipid, and the Elftman t echnique for determination of phospholipid. Material was prepared for electron microscopy by fixation for one hour in l% osmium tetroxide (Millonig, 1963), dehydration in graded acetones followed by propylene oxide, and embedding in Maraglas epoxy resin (Freeman and Spurlock, 1962). Grids prepared from these specimens were stained either with 1% phosphotungstic acid in 95% alcohol, or with lead citrate (Reynolds, 1963). They were scanned with either an R.C.A. EMU 2-B electron microscope or a Siemens Elmiskop I. [TRUNCATED] / 2031-01-01

Thesis (Ph.D.)--Boston University / The golden hamster, Mesocricetus auratus, is unique in that it frequently accepts not only homografts but even heterografts of normal and malignant tissues. One of the many theories a tterpting an explanation of this phenomenon, namely that lymphatic tissues that drain the sites of imphntation do not respond in a t;rpicol fashion, motivated this study. Thus, the weight changes ,and the c;'tolof'ical variations of the superficial cervical nodes in response to homologous and heterologous normal and malignant tissue transplants in the cheek pouch of the hamster were studied. The major objectives were: (1) to determine if there be any "defect" in the hamster's lymphatic tissue response to the various transplants; (2) to investigate the effects of the grafts on the large lymphoid cells of the cortex and on the plasma cells of the medulla; and ( 3) to investigate the feasibility of employing the histological picture of a regional node draining the site of a tumor heterotransplant as a base line for anti-tumor studies during the cortisone conditioning. [TRUNCATED]

Thesis (Ph.D.)--Boston University / PLEASE NOTE: Boston University Libraries did not receive an Authorization To Manage form for this thesis or dissertation. It is therefore not openly accessible, though it may be available by request. If you are the author or principal advisor of this work and would like to request open access for it, please contact us at open-help@bu.edu. Thank you. / Quantitative relationships between blood flow velocity, vessel diameter, width of the peripheral plasma layer and induction of thrombus formation were studied in small blood vessels of the retrolingual membrane and intestinal mesentery of the frog, Rana Pipiens, and of the cheek pouch of the golden hamster, Mesocricetus auratus. Blood flow velocity was measured by a modification of the technique described by Hugues (Arch. Int. de Physiol. 61: 565, 1953). Internal vessel diameters and widths of the total peripheral plasma layer were measured with an ocular micrometer. Thrombus thresholds were determined by graded electrical stimulation. Determinations were made on 202 vessels in retrolingual membranes of frogs prepared by single pithing. In 95 arterioles (diameters from 10 to 40 microns), the mean flow velocity was 2.84 mm/sec. and the mean volume flow rate, 1.68xl0-3 cu mm/sec. The mean width of the total peripheral plasma layer was 3.6 microns and its ratio to mean vessel diameter was 1:6. In 107 venules (diameters from 10 to 50 microns), the mean flow rate was 1.20 mm/sec. The mean volume flow rate was 1.02x10-3 cu mm/sec. The mean width of the total peripheral plasma layer was 4.1 microns and its ratio to mean vessel diameter was 1:7.4. The ratios of mean arterial to mean venous velocity and volume flow were 1:2.3 and 1:1.6 respectively. Measurements were made on 100 mesenteric vessels of frogs prepared by single pithing. In 50 arterioles (diameters from 15 to 45 microns), the mean velocity was 3.77 mm/sec. The mean volume flow was 3.05xlo-3 cu mm/sec. The mean width of the total peripheral plasma layer was 2. 7 microns and its ratio to mean vessel diameter was 1:11.5. In 50 venules (diameters from 20 to 50 microns), the mean velocity was 1.55 mm/sec. and the mean volume flow rate, 1.76x10-3 cu mm/sec. The mean width of the total peripheral plasma layer was 6.7 microns and its ratio to mean vessel diameter was 1:5.3. The ratios of mean arterial to mean venous velocity and volume flow were 1:2.3 and 1:1.7 respectively. A total of 109 mesenteric vessels were studied in frogs anesthetized with urethane. In 55 arterioles (diameters from 15 to 45 microns), the mean velocity was 3.28 mm/sec. The mean volume flow was 2.87x10-3 cu mm/sec. The mean width of the total peripheral plasma layer was 3.2 microns and its ratio to mean vessel diameter was 1:9.3. In 54 venules (diameters from 20 to 50 microns), the mean flow rate was 1.61 mm/sec. and the mean volume flow, 1.83x10-3 cu mm/sec. The mean width of the total peripheral plasma layer was 5.6 microns and its ratio to mean vessel diameter was 1:5.9. The ratios of mean arterial to mean venous velocity and volume flow were 1:2.0 and 1:1.5 respectively. In these three series of studies in the frog, no relationship was clearly apparent between velocity and either vessel diameter or width of the peripheral plasma layer in arterioles. Suggestions of a direct relationship between velocity and peripheral plasma layer in veins, however, were evident. In all cases, and in both arterioles and venules, vessel diameter and peripheral plasma layer were clearly and directly related. The effects of flow velocity changes on width of the total peripheral plasma layer in individual vessels were studied in 26 arterioles of the hamster cheek pouch. Blood flow was varied by means of a cuff described by Copley (Biorheology 1: 3, 1962). A direct relationship between velocity and width of the peripheral plasma layer was clearly demonstrated. Thrombus thresholds were determined in mesenteric vessels of the frog. In 103 arterioles (diameters from 20 to 140 microns), the mean strength of stimulus necessary to produce a platelet thrombus was 24.8 volts with an amperage of 0.18 milliamperes. In 100 venules (diameters from 20 to 140 microns), the mean strength of stimulus necessary was 20.7 volts with an amperage of 0.12 milliamperes. Possible relationships of thrombus thresholds to flow velocity in mesenteric arterioles and venules are discussed. / 2031-01-01

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O conhecimento da morfologia e ultraestrutura dos helmintos permite a correta classificação destes organismos, bem como fornece subsídios que poderão ser utilizados para diagnóstico e controle. A microscopia laser confocal é uma ferramenta para estudar a organização estrutural de várias espécies de helmintos, possibilitando acesso a detalhes morfológicos não evidenciados pela microscopia óptica. Echinostoma paraensei é um trematódeo, digenético, hermafrodita parasito de numerosos hospedeiros vertebrados. Neste trabalho foi investigado o desenvolvimento dos órgãos reprodutivo e a morfometria de E. paraensei, desde a fase jovem até a adulta, como contribuição ao conhecimento do desenvolvimento reprodutivo desta espécie. Os trematódeos foram recuperados aos 3, 4, 5, 6, 7, 10, 14 e 21 dias posterior à infecção (dpi) experimental em hamsters. Estes foram corados em carmim clorídrico, desidratados em série alcoólica e montados em lâmina permanente em bálsamo do Canadá, fotografados e medidos usando microscopia de luz de campo claro (MCC) e microscopia de varredura laser confocal (MVLC). Entre 3 e 4 dpi, os primórdios genitais estavam presentes e nenhuma organização do sistema reprodutivo foi visualizada por MCC e MVLC. Os primórdio do ovário, dos testículos e da bolsa do cirro foram visualizados por MCC aos 5 e 6 dpi, no entanto, MVLC dos helmintos aos 5dpi mostra que estes primórdios, o ootipo e o útero estavam presentes, como estruturas individualizadas. A bolsa do cirro apresenta metratermo e o ovário com primórdio do oótipo adjacente aos 7dpi por MVLC. A vesícula seminal, receptáculo seminal, células diferenciadas nos testículos, ducto e reservatório vitelínico e oviducto foram visualizados após 10 dias, enquanto os espermatozóides na vesícula seminal, ovos e oócitos, células vitelínicas, poro e canal de Laurer aos 14 dias. A morfometria evidencia um acelerado crescimento dos órgãos reprodutores a partir do 7 dia. Os testículos apresentam aumento significativo no comprimento do 7 ao 21 dia e o ovário durante o período de 7 à 10 dpi. Aos 21 dpi, todos os helmintos apresentaram glândulas vitelínicas, útero contendo ovos e espermatozóides no oviducto enquanto outros ovos estão sendo formados. As mudanças morfológicas acentuadas durante a gametogênese consistem no aumento do comprimento do helminto, maturação das gônadas, desenvolvimento e maturação das glândulas vitelínicas. O desenvolvimento do helminto como um todo está relacionado à maturação dos órgãos reprodutivo masculino e feminino indicando o investimento deste trematódeo em garantir a produção e eliminação dos ovos ao meio exterior. / The knowledge about morphology and ultra structure of helminthes are great importance in correct classification these organisms. The Scanning Laser Microscopy (LSM) is an important tool to study the structural organization of several helminthes species. Echinostoma paraensei is a trematode, digenetic, hermaphroditic parasite of several hosts. In this study, the development of reproductive organs and the morphometry of E. paraensei from young stage to adult worm were investigated, to contribute knowledge of the reproductive development of this specie. The trematodes were recovered on 3, 4, 5, 6, 7, 10, 14 and 21 days post infection (dpi) from experimental hamsters. It were dehydrated in alcohol series, stained with hydrochloric carmine, mounted on permanent slide using Canada balsam, photographed and measured using light microscopy (LM) and Scanning Laser Microscopy (LSM). Between 3 and 4 dpi the genital anlage were present and were not observed reproductive system organization by either LM and LSM. The anlage of ovary, testes and Cirruss sac were seen at 5 and 6dpi by LM, however LSM from 5dpi image shows theses anlage, ootype and uterus are present as individualized structure. Cirrus sac showed metraterm and ootype adjacent to primordial ovary were seen at 7 dpi by LSM. The seminal vesicle, seminal receptacle, differentiated cells in the testes, viteline ducts and oviduct were visualized after 10 days infection, while sperms in seminal vesicle, eggs, oocyte, Laurer canal and pore from 14 dpi by LSM. The morphometry shows a rapid growth of reproductive organs from 7th day. The testes have significantly increased length from 7 until 21dpi and ovary from 7 until 10dpi. All helminthes showed vitellines glands, uterus contained eggs and sperm in oviduct while another eggs were forming at 21 dpi. The marked morphologic changes during gametogenesis are increase of body length of helminthes, gonad maturation and development and maturation of vitelline glands. The development of helminthes as a whole is related to maturation of female and male organs of reproductive system showing the investment this trematode taken to ensuring the production, maintenance and delivery the eggs to external environment.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O conhecimento da morfologia e ultraestrutura dos helmintos permite a correta classificação destes organismos, bem como fornece subsídios que poderão ser utilizados para diagnóstico e controle. A microscopia laser confocal é uma ferramenta para estudar a organização estrutural de várias espécies de helmintos, possibilitando acesso a detalhes morfológicos não evidenciados pela microscopia óptica. Echinostoma paraensei é um trematódeo, digenético, hermafrodita parasito de numerosos hospedeiros vertebrados. Neste trabalho foi investigado o desenvolvimento dos órgãos reprodutivo e a morfometria de E. paraensei, desde a fase jovem até a adulta, como contribuição ao conhecimento do desenvolvimento reprodutivo desta espécie. Os trematódeos foram recuperados aos 3, 4, 5, 6, 7, 10, 14 e 21 dias posterior à infecção (dpi) experimental em hamsters. Estes foram corados em carmim clorídrico, desidratados em série alcoólica e montados em lâmina permanente em bálsamo do Canadá, fotografados e medidos usando microscopia de luz de campo claro (MCC) e microscopia de varredura laser confocal (MVLC). Entre 3 e 4 dpi, os primórdios genitais estavam presentes e nenhuma organização do sistema reprodutivo foi visualizada por MCC e MVLC. Os primórdio do ovário, dos testículos e da bolsa do cirro foram visualizados por MCC aos 5 e 6 dpi, no entanto, MVLC dos helmintos aos 5dpi mostra que estes primórdios, o ootipo e o útero estavam presentes, como estruturas individualizadas. A bolsa do cirro apresenta metratermo e o ovário com primórdio do oótipo adjacente aos 7dpi por MVLC. A vesícula seminal, receptáculo seminal, células diferenciadas nos testículos, ducto e reservatório vitelínico e oviducto foram visualizados após 10 dias, enquanto os espermatozóides na vesícula seminal, ovos e oócitos, células vitelínicas, poro e canal de Laurer aos 14 dias. A morfometria evidencia um acelerado crescimento dos órgãos reprodutores a partir do 7 dia. Os testículos apresentam aumento significativo no comprimento do 7 ao 21 dia e o ovário durante o período de 7 à 10 dpi. Aos 21 dpi, todos os helmintos apresentaram glândulas vitelínicas, útero contendo ovos e espermatozóides no oviducto enquanto outros ovos estão sendo formados. As mudanças morfológicas acentuadas durante a gametogênese consistem no aumento do comprimento do helminto, maturação das gônadas, desenvolvimento e maturação das glândulas vitelínicas. O desenvolvimento do helminto como um todo está relacionado à maturação dos órgãos reprodutivo masculino e feminino indicando o investimento deste trematódeo em garantir a produção e eliminação dos ovos ao meio exterior. / The knowledge about morphology and ultra structure of helminthes are great importance in correct classification these organisms. The Scanning Laser Microscopy (LSM) is an important tool to study the structural organization of several helminthes species. Echinostoma paraensei is a trematode, digenetic, hermaphroditic parasite of several hosts. In this study, the development of reproductive organs and the morphometry of E. paraensei from young stage to adult worm were investigated, to contribute knowledge of the reproductive development of this specie. The trematodes were recovered on 3, 4, 5, 6, 7, 10, 14 and 21 days post infection (dpi) from experimental hamsters. It were dehydrated in alcohol series, stained with hydrochloric carmine, mounted on permanent slide using Canada balsam, photographed and measured using light microscopy (LM) and Scanning Laser Microscopy (LSM). Between 3 and 4 dpi the genital anlage were present and were not observed reproductive system organization by either LM and LSM. The anlage of ovary, testes and Cirruss sac were seen at 5 and 6dpi by LM, however LSM from 5dpi image shows theses anlage, ootype and uterus are present as individualized structure. Cirrus sac showed metraterm and ootype adjacent to primordial ovary were seen at 7 dpi by LSM. The seminal vesicle, seminal receptacle, differentiated cells in the testes, viteline ducts and oviduct were visualized after 10 days infection, while sperms in seminal vesicle, eggs, oocyte, Laurer canal and pore from 14 dpi by LSM. The morphometry shows a rapid growth of reproductive organs from 7th day. The testes have significantly increased length from 7 until 21dpi and ovary from 7 until 10dpi. All helminthes showed vitellines glands, uterus contained eggs and sperm in oviduct while another eggs were forming at 21 dpi. The marked morphologic changes during gametogenesis are increase of body length of helminthes, gonad maturation and development and maturation of vitelline glands. The development of helminthes as a whole is related to maturation of female and male organs of reproductive system showing the investment this trematode taken to ensuring the production, maintenance and delivery the eggs to external environment.

Leishmaniasis are zoonosis with different clinical manifestations according to the Leishmania species involved and the host immune response. Leishmania braziliensis complex is responsible for the majority of human Tegumentary Leishmaniasis cases in Brazil and it is endemic in the Triângulo Mineiro region. Our aim was to compare the pathogeny of two different isolates of Leishmania braziliensis complex, obtained from cutaneous lesions of patients from Triângulo Mineiro region, with Leishmania (Viannia) braziliensis reference strain (MHOM/BR/75/M2903). Comparision was done by histopathological and ultrastructural analisys of organs of experimentally infected Hamster golden (Mesocricetus auratus). For that, 106 stationary promastigotes of the two isolates and the reference strain were subcutaneously innoculated in the hind forepaw of Hamster that were killed after 15, 30, 60 e 120 days post infection and then were collected fragments of the skin, popliteal lymph node, liver, spleen and bone marrow. The organs were processed for histopathological and ultrastructural analysis. The results showed differents pathogeny between both isolates and the reference strain, characterized for differences in prepatent period of the macroscopy lesions, in the outcome of the histophatological lesions, in the parasitism and in the capacity to produce metastatic lesions in the lymph node, liver and spleen. / As leishmanioses são zoonoses que produzem diferentes manifestações clínicas dependendo da espécie de Leishmania envolvida e da resposta imune do hospedeiro. O complexo Leishmania braziliensis é responsável pela maioria dos casos da leishmaniose tegumentar humana no Brasil e é endêmica na região do Triângulo Mineiro. Este trabalho teve como objetivo comparar a patogenia de dois diferentes isolados do complexo Leishmania braziliensis, provenientes de lesões cutâneas de pacientes da região do Triângulo Mineiro (MG), com a cepa de referência Leishmania (Viannia) braziliensis MHOM/BR/75/M2903, através de análise histopatológica e ultra-estrutural de órgãos de Hamster golden (Mesocricetus auratus) infectados experimentalmente. Para isto, 106 promastigotas na fase estacionária de desenvolvimento de cada isolado e da cepa de referência foram inoculadas em coxim plantar de Hamsters, os quais, após períodos de 15, 30, 60 e 120 dias de infecção, foram sacrificados e destes retirados fragmentos de pele, linfonodo poplíteo, fígado, baço e medula óssea. Os órgãos foram processados para avaliação histopatológica e ultra-estrutural. Os resultados obtidos dessas análises revelaram diferentes patogenias entre os dois isolados e a cepa de referência, caracterizadas por diferenças no período de prepatência das lesões macroscópicas, no aparecimento das lesões histopatológicas, no parasitismo e na capacidade de produzir lesões metastáticas no linfonodo, fígado e baço. / Mestre em Ciências da Veterinárias

The main aim of this thesis was to prove the hypothesis that the intake of blood with specific antibodies against sand fly saliva affects sand fly fecundity and mortality. Phlebotomus argentipes and Mesocricetus auratus were used for most experiments. ELISA revealed high levels of specific antibodies in repeatedly exposed hosts. However, a five-day study of mortality and fecundity of bloodfed females demonstrated that feeding on repeatedly bitten hamsters has no effect on number of eggs or survival of females. Salivary antigens of P. argentipes recognized by sera of repeatedly bitten hamsters were characterized by immunoblotting and mass spectrometry. Immunoblotting showed that antibodies in the hamster sera recognize salivary proteins with molecular weight from 25 to 60 kDa. Mass spectrometry revealed that the antigens correspond to D7, apyrases, antigen 5-related proteins and yellow-related proteins. In addition, Phlebotomus females were fed through a chicken membrane on rabbit blood with high concentrations of histamine, serotonin or prostaglandin E2 to find out if they influence fecundity or mortality of sandfly females. While the approximate number of eggs layed by one female did not significantly differ from controls in any experimental group studied, the total mortality of females was lower...

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