Este sumário apresenta o resumo e os resultados deste estudo experimental do Fechamento do Coto Duodenal, em um e dois planos de sutura, realizado em 48 cães submetidos a uma gastrectomia Billroth II, com a finalidade de estudar comparativamente, o processo de cicatrização, macro e microscópico da sutura deste Coto. Os aspectos mais relevantes deste estudo de cicatrização de feridas macroscópicas e microscópicas no coto duodenal foram os seguintes: A) Estudo macroscópico: Com o objetivo de realizar um estudo macroscópico de fechamento do coto duodenal, foi realizada uma gastrectomia BII, em 48 cães, divididos em dois grupos: Grupo 1 - composto por 24 cães em que a sutura do coto duodenal foi realizada em um plano extra-mucoso. Estes animais foram divididos em dois subgrupos de 12 animais cada: i1 - cães sacrificados no período pós-operatório imediato, ou seja, no primeiro, quarto e sétimo dia após a operação; e M1 - cães sacrificados em um período pós-operatório mais tardio, ou seja, no décimo quarto, vigésimo primeiro e vigésimo oitavo dia, após a operação. Grupo 2 - Composto por 24 animais, em que a sutura do coto duodenal foi realizada em 2 planos (total e seromuscular). Também estes animais foram distribuídos em subgrupos, i2 e M2, de forma idêntica como no Grupo 1. Um estudo comparativo foi feito quanto aos resultados de fechamento do coto duodenal em 1 e 2 planos, levando em consideração seus aspectos macroscópicos, obtendo-se as seguintes conclusões: o Em nenhum dos cães, em ambos os grupos, ocorreu deiscência do coto duodenal ou quaisquer outras complicações pós-operatórias; o O diâmetro externo do coto duodenal ao nível da sutura foi significativamente maior no grupo com 2 planos de sutura. No entanto, a 5 e 10 cm distalmente à sutura do coto, esta medida foi semelhante em ambos os grupos. o Aderências ao coto duodenal foram semelhantes em ambos os métodos de sutura, exceto por uma incidência significativamente maior de aderências de epíplon no grupo com um plano de sutura, durante os primeiros 7 dias. o A inflamação da mucosa foi significativamente maior no grupo com 2 planos de sutura, durante os primeiros 7 dias; o O esporão formado no fechamento do coto em 2 planos de sutura foi significativamente maior que o formado em um plano de sutura. o A distância entre a superfície mucosa do coto duodenal e a papila maior e papila menor do duodeno, foi significativamente maior nos cotos fechados com 1 plano de sutura. B) O estudo histológico: Um estudo histológico de fechamento do coto duodenal foi feito, após gastrectomia Billroth II em 48 cães, divididos em dois grupos iguais. No Grupo 1, o fechamento foi realizada em um plano, enquanto no Grupo 2 foi feito em dois planos (total e seromuscular). Em cada grupo, os animais foram divididos em dois subgrupos; i1 - cães sacrificados no início do período pós-operatório, ou seja, no primeiro, quarto e sétimo dias; e M1 - cães sacrificados no pós-operatório tardio, ou seja, no décimo quarto, vigésimo primeiro e vigésimo oitavo dia. Não houve um único caso de deiscência em qualquer animal. As conclusões relativas aos aspectos histológicos mostraram os seguintes aspectos: o Houve um esporão invertido para o lúmen do intestino, em todos os animais do grupo 2, enquanto que um pseudoesporão foi observado em apenas 37% nos animais do Grupo 1 ; o Sinais inflamatórios agudos foram mais intensos no grupo 2, na primeira semana de pós-operatório; o A formação de neofibrose do tecido conjuntivo e a regeneração da \"muscularis mucosae\" foi semelhante nos dois grupos. No entanto, a regeneração da mucosa ocorreu mais precocemente no grupo 1 , ao passo que a regeneração das fibras musculares ocorreu mais precocemente no grupo 2 / An experimental study of duodenal stump closure in one and two suture layers was performed in 48 dogs submitted to a Billroth II Gastrectomy, in order to study comparatively Macroscopic and Histological wound healing aspects. The most relevant aspects of this Macroscopic and Histological wound healing study in the duodenal stump, were the following ones: A) Macroscopic study: A comparative study was made as to the results of duodenal stump closure in 1 and 2 layers, taking in consideration its macroscopic aspects. Billroth II gastrectomy was performed in 48 dogs divided in two groups: a. Group 1 - consisted of 24 dogs in which the duodenal stump suture was performed in one extra-mucous layer. These animals were distributed into 2 subgroups of 12 animals each: i1 - dogs sacrificed in the early postoperative period, that is, on the first, fourth and seventh day after the operation; and M1 - dogs sacrificed at a later postoperative period, that is, on the fourteenth, twenty-first and twenty-eighth day after the operation. b. Group 2 - consisted of 24 dogs, in which the suture of the duodenal stump was performed in 2 layers (total and seromuscular ones). Also these animals were distributed in i2 and M2 sub-groups, identically as in group 1. The following conclusions were obtained, concerning the macroscopic aspects: o Dehiscence of the duodenal stump or any other postoperative complications occurred in none of the dogs in both groups; o The external diameter of the duodenal stump at the level of the suture was significantly larger in the group with 2 suture layers. However, at 5 and 10 cm distally from the suture, this measure was similar in both groups. o Adherences to the duodenal stump were similar in both suturing methods, except for a significantly higher incidence of epiplon adherence in the 1 layer group during the first 7 days. o Mucosal inflammation was significantly greater in the 2 suture layers group during the first 7 days; o The spurs formed in the 2 layers stump closure was significantly larger than that formed in 1 layer suture. o The distance from the mucosal surface of the stump suture to the Major and Minor papilla of the duodenum was significantly greater in the stumps closed with 1 layer sutures. B) Histological study: A histological study of duodenal stump closure was made, following a Billroth II gastrectomy in 48 dogs, divided in two equal groups (1 and 2) and its subgroups (i1 and M1; i2 and M2), as mentioned above. The conclusions concerning to histological aspects showed the following characteristics: o There was an inverted spur toward the lumen of the bowel, in all group 2 animals, whereas a pseudo- spur was also observed in only 37% in group 1 animals; o Acute inflammatory signs were more intense in group 2 in the first week of recovery; o The formation of young connective tissue fibrosis and muscularis mucosae fibers regeneration was similar in both groups. However, mucosal regeneration occurred earlier in group 1, while regeneration of muscle fibers occurred more precociously in group 2

Visando avaliar a atuação dos biorreguladores beta-NOA, AIB e GA3 na morfogênese de microplantas de abacaxi Ananas comosus L. (Merrill) cultivar IAC Gomo-de-Mel estabeleceu-se um experimento com a presença isolada e/ou combinada dos respectivos biorreguladores ao meio de cultura MS. As microplantas foram mantidas em condições de cultura in vitro, com temperatura, luminosidade e fotoperíodo controlados (25 ± 2 C, irradiância de 42 µmol.m-2s-1 e 16 horas, respectivamente) durante 3 meses. Os resultados obtidos com a análise de variância evidenciaram que houve interação do biorregulador de crescimento e o tempo de cultivo para a altura, número de raiz e número de folhas. O beta-NOA induziu a rizogênese foliar nas microplantas a partir de células da bainha do feixe vascular e não apresentou toxicidade nas concentrações utilizadas, ao passo que o AIB mostrou-se mais eficaz para a indução da rizogênese adventícia caulinar. Em relação ao crescimento das microplantas, a ação tanto isolada, como combinada em ambas as concentrações avaliadas de GA3 foram mais eficientes. As análises histológicas evidenciaram alterações no número de estratos da epiderme, hipoderme e mesofilo das folhas cultivadas nos tratamentos com biorreguladores, porém não se verificou alterações citológicas em nenhum dos tratamentos. Acredita-se que a utilização de pulsing seja uma alternativa eficaz para a rizogênese e crescimento em altura das microplantas de abacaxizeiro Gomo-de-mel. / In order to evaluate the action of bioregulators beta-NOA, IBA and GA3 in the pineapple Ananas comosus L. (Merril) cultivar IAC gomo-de-mel microplants morphogenesis an experiment was stablished in order to the presence of the bioregulator alone and in combinations. The microplants were kept in culture in vitro conditions, during 3 months under controlled temperature, light and photoperiod (25 ± 2 C, irradiance de 42 µmol.m- 2s-1 e 16 hours). The analysis of variance showed the interaction interaction between the use of bioregulator and the period of cultivation on hight, number of roots and leaves. The beta-NOA induced leaf rhizogensis in the microplants from the bundle sheath and there was no sign of toxicity with the used concentrations. The IBA showed to be more effective in the adventicious steam rhizogenesis. The isolated and combinated actions of GA3 were the most effective treatment to microplants growing. The histological analysis showed alterations on the epidermis, hypodermis and mesophyll number of layers of the cultivated leaves with this bioregulator, but neither of the treatments showed citological alterations. It is believed that the use of pulsing would be an effective alternative for the rhizogenesis and growing of the gomo de mel pineapple microplants.

Psoriasis is a chronic, clinically heterogeneous, skin condition that affects approximately 2% of the general population. In 1985, Henseler and Christophers, classified psoriasis into early onset (EOP; age at onset ≤40 years-y) and late onset disease (LOP; age at onset >40 y). Previous research suggests that there are genetic and immunological differences between EOP and LOP. In particular, the major genetic determinant for psoriasis, the human leukocyte antigen (HLA)-Cw6 allele, occurs more frequently in EOP (55-80%) compared to LOP (15-20%) patients. Epidermal Langerhans’ cells (LC) migration is also different in these 2 subtypes of psoriasis. The primary aim of this thesis was to further explore the clinical, histological and immunohistochemical (IHC) differences between EOP and LOP. We compared clinical characteristics in a total of 497 subjects, including 340 psoriasis patients (108 recruited prospectively; 76 EOP and 32 LOP, mean age of onset 20.3±9.9 and 55.6±7y respectively, and 232 retrospectively; 202 EOP and 30 LOP, mean age of onset 20.7±9.9 and 55.2±7.2y respectively) and 157 controls (mean age 66±11.2y). Information on demographics, family history of psoriasis, clinical features, treatment and co-morbidities were recorded. Patients were also assessed for health-related quality of life and psychological distress. A total of 31 psoriasis patients, ≥ 50y of age, participated in the histological and IHC evaluation; 17 EOP and 14 LOP, mean age of onset 21.1±8.5 and 55.4±7.7y respectively. Skin biopsies were taken from involved (PP) and uninvolved (PN) skin and stained with haematoxylin and eosin (H&E) and IHC antibodies against various T-cell (CD3, CD4, and CD8) and LC (CD1α) markers. The H&E parameters (morphological and inflammatory) were graded with the use of a study specific histological score, whilst IHC positive epidermal cells were counted per microscopic field at 200X magnification. The dermal IHC infiltrate was assessed with a semi-quantitative (0-3) scale. Gender, body mass index, disease duration and severity, diagnosed hypertension and dyslipidemia were treated as covariates. The clinical data showed that LOP patients had a lower likelihood of having a positive family history of psoriasis (62% of EOP versus 35.6% of LOP patients; chi square-x2, P=0.001). In addition, patients with EOP parent(s) were 91% less likely to develop LOP than EOP (odds ratio-OR=0.093, P=0.025, 95% confidence interval-CI 0.012-0.74). Moreover, compared to LOP, EOP patients had a more severe disease (x2, P=0.021), usually requiring 3rd line treatments (x2; P=0.010). They also experienced frequent flares, following upper respiratory tract infections (x2, P=0.049). When data were segregated by age (≥50years) and after accounting for covariates, we observed that, compared to the non-psoriasis population, LOP patients were approximately 3 times more likely to develop type 2 Diabetes Mellitus (OR=2.56, P=0.05, 95% CI 1.01-6.54), whilst, EOP subjects were 98% less likely to develop autoimmune thyroiditis (OR=0.025, P=0.02, 95% CI 0.001-0.55). Psychologically, LOP patients were found to be a clinically more anxious group compared to EOP (t-test, P=0.006). Microscopically, the results from the H&E study showed an increased total inflammatory infiltrate in LOP, PP sections compared to EOP, PP ones (t-test, P=0.028). With IHC stains, we observed that in the epidermis of LOP PP, there was a significantly higher count of CD4+ cells; mean CD4+ in LOP of 15.1 ± 6.2 versus 6.7±4.6 in EOP (Analysis of variance-ANOVA, P<0.001). This subsequently led to a higher epidermal CD4+/CD8+ ratio of 1.3 in the LOP versus 0.5 for the EOP sections (ANOVA, P=0.002). In the PP dermis, CD4+ were also more abundant in the LOP tissue (x2, P=0.049). To assess whether these CD4+ cells were either T-lymphocytes or LC, we examined for differences in the CD3+ and CD1α+ cells. The mean epidermal CD3+ tended to be higher in LOP PP sections; mean epidermal CD3+ in the LOP 42.8 ± 13.3 versus 31.7 ± 17.5 in the EOP group (ANOVA, P= 0.061), while the dermal infiltrate showed a similar pattern (x2,P=0.067). Finally, there was no difference in epidermal and dermal CD1α+ and CD8+ cells in PP between EOP and LOP sections. These data indicate differences in clinical phenotype, heritability, comorbidities and immunopathomechanism between EOP and LOP. Taken together they provide further evidence that EOP and LOP may be different diseases.

O petróleo é um dos xenobióticos mais utilizados pela humanidade e também um dos mais prejudiciais aos seres vivos. A formação do petróleo é composta em 97% por hidrocarbonetos, que são os maiores poluentes em potencial. Os hidrocarbonetos policíclicos aromáticos (HPAs), os maiores constituintes do petróleo, são poluentes orgânicos de grande persistência ambiental, e seus compostos derivados são potenciais carcinogênicos que podem afetar toda biota em que estejam envolvidos. O presente estudo teve como objetivo avaliar os efeitos da exposição subletal aos HPAs na concentração de 0,4 ppm em peixes marinhos da espécie Rachycentron canadum por meio de parâmetros morfométricos, metabólicos e histológicos. Os HPAs foram obtidos através da extração da fração solúvel de petróleo em água (FSA) e avaliados qualitativamente por meio de espectrometria de fluorescência. Foram avaliados os índices hepato-somáticos (IHS), assim como as possíveis alterações histológicas do tecido hepático dos peixes, supostamente decorrentes da exposição ao xenobiótico. Os parâmetros metabólicos através de biomarcadores de estresse como o cortisol, a glicose e o lactato plasmáticos, assim como a análise complementar por meio das proteínas totais foram determinados. Os resultados obtidos neste trabalho demonstram que os espécimes de Rachycentron canadum não sofreram alterações nos parâmetros estressores quando submetidos à exposição aos HPAs. Quanto à análise histológica, os animais experimentais apresentaram alterações moderadas a graves no fígado decorrente da exposição aos HPAs. Os beijupirás demonstraram ser bons bioindicadores, por suas características comportamentais e respostas aos parâmetros analisados, o que pode viabilizá-lo como objeto de pesquisa. Cada vez mais, estudos que avaliam o impacto de xenobióticos como os HPAs, os quais em concentrações mínimas em organismos aquáticos podem afetar todo o equilíbrio da biota, evidenciam a importância do monitoramento ambiental, uma vez que a dispersão dos xenobióticos no ambiente pode resultar em mudanças em longo prazo. / Oil is one of the humanity most used xenobiotics, and also one of the more dangerous to animals. Oil has about 97% of hydrocarbons in its constitution, which have the major pollutant potential. Polycyclic Aromatic Hidrocarbons (PAHs) are organic pollutants of high environmental persistence level, and its derivatives have carcinogenic potential and can affect all involved biota. The present study aimed to evaluate the sub lethal exposure effects do PAH in the marine fish Rachycentron canadum by morphometric, metabolic and histological parameters. PAHs were obtained by extraction of oil soluble fraction in water (WSA) and were quantitatively evaluated by fluorescence spectrometry. Bioassays were performed using the concentration of 0.4 ppm previously determined. We evaluated the hepatosomatic index (HSI) and the possible histological changes in the liver of the fish, supposedly obtained in consequence of the exposure to the xenobiotic. Metabolic parameters using biomarkers of stress such as cortisol, plasma glucose and lactate, as well as additional analysis by total protein, were determined. Our results showed that Rachycentron canadum did not show alterations on the parameters of stress. Regarding to the histological analysis, animals presented mild to severe changes in the liver, possibly as result of PAHs exposure. Rachycentron canadum resulted to be a good bioindicator, because of its behavioral characteristics and responses to analyzed parameters, which can make it viable as subject for researches. Studies that evaluate xenobiotic impacts like PAHs, in which minimal concentrations can affect the equilibrium of all biota, show the importance of environmental monitoring, once environmental xenobiotics dispersion can produce long term changes.

Magister Chirurgiae Dentium (MChD) / The author is interested in the morphogenesis of the oral region including the nasopalatine complex. With the intention of undertaking a study of the· embryological development in this area, perusal of available literature failed to reveal a single compreh.ensive description of the reception and handling of embryonic and foetal material, mensuration and preparation for miscroscopy. Human material for embryological study is relatively scarce in· the Republic of South Africa. According to the literature there is, however, a distinct similarity between human and domestic pig development in certain regions, notably the palate. Furthermore, pig embryos and foetuses are available in comparative abundance from sows slaughtered at abattoirs. As a consequence of the above-mentioned factors it was,decided to undertake a -preparatory study in order to firstly evaluate existing methods of handling of embryonic and foetal material and secondly, to statistically evaluate data relating to mass and measurements.'· The aim was to draw a comparison with existing information and to select a sample for investigation. Embryos and foetuses were removed from slaughtered sows in a fresh state and removed to the laboratory immersed in 10 per cent neutral buffered formol saline. In the laboratory foetal membranes were removed, umbilical cords cut and the specimens weighed. They were then placed in Bouin's solution for final fixation and decalcification. Instruments were designed to measure crown-tailroot length, crown-rump length and dorsal profile length. After one day in Bouin's solution all specimens were measured. In order to determine the accuracy of the weighing and measuring procedures ten fixed specimens were weighed and measured on seven consecutive days. Statistical analysis of this data indicated that crown-rump length was the most accurately determinable linear measurement, judged by both the coefficient of variation and the standard deviation. On this basis crown-rump length was chosen as the criterion for selecting the sample to be studied. Correlation between linear measurements and between linear measurements and mass for the entire series showed a very strong positive relationship between all the parameters indicating that a dimensional relationship was maintained during growth. After measuring, the small specimens were embedded whole while larger embryos and foetuses were decapitated. A method was described for trimming and embedding these heads in such a way that subsequent sectioning would take place in a standardised transverse plane. In larger specimens this procedure had to be delayed until demineralization had taken place. Conclusions based on a consideration of data for the entire population included the following: 1. The mean number of specimens per litter was 6,475. 2. The number of pigs per litter stayed relatively constant throughout the period of gestation. 3. Mass showed a greater intra-litter variation than any of the three linear measurements recorded. 4. Relatively, lengths appeared to vary less in older than in younger Ldtt.ers-, irrespective of litter size

Bladder cancer (BCa) is the fourth most commonly diagnosed cancers in men and the eighth most common in women. It is an abnormal growth of tissues which develops in the bladder lining. Histological analysis of bladder tissue facilities diagnosis as well as it serves as an important tool for research. To bet- ter understand the molecular profile of bladder cancer and to detect predictive and prognostic features, microscopy methods, such as immunofluorescence (IF), are used to investigate the characteristics of bladder cancer tissue. For this project, a new method is proposed to segment cancer epithelial us- ing nuclei morphology captured with IF staining. The method is implemented using deep learning algorithms and performance achieved is compared with the literature. The dataset is stained for nuclei (DAPI) and a marker for cancer epithelial (panEPI) which was used to create the ground truth. Three popu- lar Convolutional Neural Network (CNN) namely U-Net, Residual U-Net and VGG16 were implemented to perform the segmentation task on the tissue mi- croarray dataset. In addition, a transfer learning approach was tested with the VGG16 network that was pre-trained with ImageNet dataset. Further, the performance from the three networks were compared using 3fold cross-validation. The dice accuracies achieved were 83.32% for U-Net, 88.05% for Residual U-Net and 82.73% for VGG16. These findings suggest that segmentation of cancerous tissue regions, using only the nuclear morphol- ogy, is feasible with high accuracy. Computer vision methods better utilizing nuclear morphology captured by the nuclear stain, are promising approaches to digitally augment the conventional IF marker panels, and therefore offer im- proved resolution of the molecular characteristics for research settings.

Technological advancements have opened up the possibility of digitizing the pathological landscape, enabling deep learning-based methods to analyze digitized tissue samples, i.e., whole slide images (WSIs). Attention has recently shifted toward modeling WSIs as graphs since graph representations can capture dynamic relationships. This thesis investigates different graph construction techniques in conjunction with graph-based deep learning to classify WSIs as breast cancer histological grade 1 versus histological grade 3. To that extent, multiple graph representation techniques and two graph convolutional networks, GCN and GraphSAGE, were utilized. Finally, by evaluating the proposed models on an external test set originating from a separate cohort, it is clear that both models have the capacity for binary histological grading, yielding AUC scores of 0.791 (95% CI 0.756 − 0.825) and 0.838 (95% CI 0.808 − 0.869) for the GCN and GraphSAGE models. Modeling WSIs as graphs is an exciting and emerging field; however, further work is needed to evaluate alternative graph representation techniques and graph convolutional networks. / Teknologiska framsteg har öppnat upp möjligheten att digitalisera det patologiska landskapet, och möjliggjort djupinlärningsbaserade metoder att analysera digitala vävnadsprover; whole slide images (WSIs). Uppmärksamhet har skiftats mot modellering av WSIs som grafer, då grafrepresentationer är dynamiska. Den här uppsatsen undersöker diverse tekniker för grafkonstruktion tillsammans med grafbaserad djupinlärning för att klassificera WSIs som histologisk gradering 1 kontra histologisk gradering 3. För detta ändamål användes flertalet olika grafrepresentationer samt två distinkta grafbaserade faltningsnätverk, GCN och GraphSAGE. Slutligen, genom att evaluera de föreslagna modellerna på ett externt test set, härstammande från en separat kohort, så är det tydligt att båda modellerna har kapacitet för binär histologisk gradering, med AUC-värden på 0.791 (95% CI 0.756 − 0.825) samt 0.838 (95% CI 0.808 − 0.869) för GCN och GraphSAGE. Att modellera WSIs som grafer är ett spännande och framväxande område. Det behövs emellertid mer forskning för att evaluera alternativa tekniker för grafrepresentationer samt grafbaserade faltningsnätverk.

No / The hair shaft has increasing importance in bioarchaeology, since it is now possible to retrieve detailed biomolecular information on recent life history using individual fibres (e.g., on diet, drug use and DNA). Data on hair condition is an important cornerstone to ensuring that reliable information is obtained. The following study defines morphological features of degradative change in human terminal scalp hair using different microscopy techniques. Evidence of degradative change is translated into a ranked histology for assessing hair sample condition. The approach is applied to samples of cut modern scalp hair subjected to degradation under soil burial/simulated grave conditions.

京都大学 / 新制・課程博士 / 博士(医学) / 甲第25186号 / 医博第5072号 / 京都大学大学院医学研究科医学専攻 / (主査)教授 安達 泰治, 教授 森本 尚樹, 教授 羽賀 博典 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

This thesis is concerned with the topic of teaching histology at secondary schools. Histology is focused on the study of microscopic structure of tissue. By this it comprises the basic foundation stone for the studies of organs and organ systems not merely in humans, but also in other animals. The method of preparing the permanent histological slides, which has also been tested in practice, is thoroughly described in this thesis. The main goal of the practical part was to prepare a representative collection of the permanent histological sections through the organs of the house mouse (Mus muculus), to acquire its photo documentation as a base for an interactive histological atlas. House mouse (Mus musculus) was chosen as the model animal, because of its frequent utilization in various laboratory experiments and because this model mammal has the structure of tissues similar to human. The atlas consists of the photo documentation of histological sections through the following organs: lungs, skin, heart, thymus gland, spleen, epididymis, testicle, penis, spermatic sacs, striated muscle, heart muscle, smooth muscle, liver, tongue, pancreas, small intestine, large intestine, gall bladder, kidney, urinary bladder, urethra, cerebellum and hippocampus. The sections are stained with application of...