abstract: Finger motion and hand posture of six professional clarinetists (defined by entrance into or completion of a doctorate of musical arts degree in clarinet performance) were recorded using a pair of CyberGloves® in Arizona State University's Center for Cognitive Ubiquitous Computing Laboratory. Performance tasks included performing a slurred three-octave chromatic scale in sixteenth notes, at sixty quarter-note beats per minute, three times, with a metronome and a short pause between repetitions, and forming three pedagogical hand postures. Following the CyberGloves® tasks, each subject completed a questionnaire about equipment, playing history, practice routines, health practices, and hand usage during computer and sports activities. CyberGlove® data were analyzed to find average hand/finger postures and differences for each pitch across subjects, subject variance in the performance task and differences in ascending and descending postures of the chromatic scale. The data were also analyzed to describe generalized finger posture characteristics based on hand size, whether right hand thumb position affects finger flexion, and whether professional clarinetists use similar finger/hand postures when performing on clarinet, holding a tennis ball, allowing hands to hang freely by the sides, or form a "C" shape. The findings of this study suggest an individual approach based on hand size is necessary for teaching clarinet hand posture. / Dissertation/Thesis / D.M.A. Music 2011

Meiotic recombination is a fundamental biological process in sexually reproducing organisms, enabling offspring to inherit novel combinations of mutations, and ensuring even segregation of chromosomes into gametes. Recombination is initiated by programmed Double Strand Breaks (DSBs), the genomic locations of which are determined in most mammals by PRDM9, a rapidly evolving DNA-binding protein. In crosses between different mouse subspecies, certain Prdm9 alleles cause infertility in hybrid males, implying a critical role in fertility and speciation. Upon binding to DNA, PRDM9 deposits a histone modification (H3K4me3) typically found in the promoters of expressed genes, suggesting that binding might alter the expression of nearby genes. Many other questions have remained about how PRDM9 initiates recombination, how it causes speciation, and why it evolves so rapidly. This body of work investigates these questions using complementary experimental and analytical methodologies. By generating a map of human PRDM9 binding sites and applying novel sequence analysis methods, I uncovered new DNA-binding modalities of PRDM9 and identified sequence-independent factors that predict binding and recombination outcomes. I also confirmed that PRDM9 can affect gene expression by binding to promoters, identifying candidate regulatory targets in meiosis. Furthermore, I showed that PRDM9’s DNA-binding domain also mediates strong protein-protein interactions that produce PRDM9 multimers, which may play an important functional role. Finally, by generating high-resolution maps of PRDM9 binding in hybrid mice, I provide evidence for a mechanism to explain PRDM9-mediated speciation as a consequence of the joint evolution of PRDM9 and its binding targets. This work reveals that PRDM9 binding on one chromosome strongly impacts DSB formation and/or repair on the homologue, suggesting a novel role for PRDM9 in promoting efficient homology search and DSB repair, both critical for meiotic progression and fertility. One consequence is that PRDM9 may play a wider role in mammalian speciation.

Made available in DSpace on 2015-07-22T20:50:03Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-11-25. Added 1 bitstream(s) on 2015-08-11T03:26:32Z : No. of bitstreams: 1 Publico-549%20-%2011769.pdf: 1027011 bytes, checksum: 549288ebc09400ab5c317f529c1f6f07 (MD5) / Objetivo: Estabelecer parâmetros da superfície palmar para localização precisa do limite proximal da polia flexora A1, nos dedos indicador, médio, anular e mínimo, e avaliar o comprimento dessa polia. Métodos: Foram estudados 280 dedos de 70 mãos de 35 cadáveres frescos, mensurando-se inicialmente a distância entre as pregas digitopalmar e interfalangeana proximal (medida A), seguindo-se com a dissecação dos dedos e mensuração da distância entre o limite proximal da polia A1 e a prega digitopalmar (medida B), bem como o comprimento da polia A1 (medida C), em cada dedo. A análise estatística foi feita com a utilização do teste multivariado “T²” de Hotelling e do teste “t” para amostras pareadas. Resultados: Após a análise estatística, foi constatado que não há diferença significante entre as medidas A e B em cada dedo (p-valor > 0,05). Também não se evidenciou diferença significante (p-valor > 0,05) entre as medidas correspondentes para cada dedo de ambas as mãos (AxA), (BxB) e (CxC). Foi observado que o comprimento médio da polia A1 em milímetros no dedo indicador é de 9,83; no médio, de 10,71; no anular, de 9,66, e, no mínimo, de 8,06. Conclusões: A distância entre as pregas digitopalmar e interfalangeana proximal deve ser usada como referência cutânea da superfície palmar para a localização precisa do limite proximal da polia A1 na palma da mão, dando mais segurança na realização de procedimentos cirúrgicos como a liberação percutânea do “dedo em gatilho”. / Purpose: We conducted this study to establish palmar surface parameters to allow for the exact location of the proximal limit of the flexor tendon A1 pulley in the index, middle, ring and little fingers and to evaluate the length of this pulley. Methods: We studied 280 fingers on 70 hands of 35 fresh human cadavers, initially measuring the distance between the digital-palmar and proximal interphalangeal creases (measure A), followed by dissection of the fingers and measurement of the distance between the proximal limit of the A1 pulley and the digital-palmar crease (measure B) and the length of the A1 pulley (measure C). A statistical analysis was carried out using Hotelling’s multivariate T2 test and the paired samples t-test. Results: No statistically significant difference was found between measures A and B in each finger (p > 0.05). The average length of the A1 pulley, in millimeters, was 9.83 in the index finger, 10.71 in the middle finger, 9.66 in the ring finger, and 8.06 in the little finger. Conclusions: The distance between the digital-palmar and proximal interphalangeal creases should be used as a cutaneous reference of the palmar surface for the exact location of the proximal limit of the A1 pulley in the palm of the hand, ensuring greater safety in surgical procedures such as percutaneous release of trigger finger. / TEDE / BV UNIFESP: Teses e dissertações

The object of this work is to chart an educational process of ambulant patients undergoing a planned arm surgery due to carpal tunnel syndrome or trigger finger and creating a functional educational material that leads to a change of knowledge and skills of those patients and that can be used by nurses during their work. The three questions to achieve the objectives were: 1) How does the educational process of the patient look like before arm surgery? 2) How is the patient educated on post-operative regime? 3) How are the educational activities implemented in patients undergoing arm surgery? The research project was aimed to the qualitative investigation. Generally the research survey was structured into two phases, the results are summarized in the practical part. The first phase was focused on observation of professional object and depth semi-structured interviews with patients who underwent an arm surgery were made. This survey was supplemented by direct structured observation of nurses in the process of educational activities. Six nurses were altogether involved in the survey. In the second phase, the educational material based on the obtained results, was made for the nurses to use during their work. This material includes educational cards and maps of care instructions for given diseases. Effectivnes of this material was subsequently verified by more semi-structured interviews with six patiens. The results of the survey showed that in the workplace where the respondents underwent the surgery, the educatinal process was singificantly eliminated and for its realization the nurses did not have satisfactory conditions, especially time and spatil conditions. During the educatinal process the nurses were using monologues teaching methods. Demonstration and practical methods were missing. Nurses were not using teaching aids that can make the educatinal process more effective. The results of this work can elucidate the educational reality in surgeries and focus attention on this sector, often neglected in expert literature.

The degree of chromatin compaction plays a fundamental role in controlling the accessibility of DNA to the transcription machinery as well as other DNA-dependent biological pathways. The mammalian HP1 (Heterochromatin protein 1) protein family consists of three members: HP1α, β and γ. Each paralogue regulates formation and maintenance of heterochromatin by binding to the repressive chromatin marks H3K9me2/3 with their chromodomains (CDs). Despite high sequence conservation, each HP1 paralogue possesses specific functions, which are likely to be cell type specific. The aim of my thesis was to find novel functions for HP1γ in mouse embryonic stem cells (mESCs) and breast cancer cells. Mass spectrometry analysis identified citrullination of residues R38 and R39 within the CD of HP1γ. I show that these residues are citrullinated by peptidyl arginine deiminase 4 (PADI4) in vitro and in vivo. Mutations in HP1γ (R38/9A), designed to mimic the loss of charge accompanied with citrullination, affect HP1γ’s binding to H3K9me3 peptides and reduce its residence time on chromatin in differentiated mESCs, indicating a role for citrullination in regulating HP1γ binding to chromatin during differentiation. Furthermore, I studied the phenotype of HP1γ depletion in two human breast cancer models and found that HP1γ is essential for cell proliferation and viability of cancer, but not of normal epithelial cells. I performed whole transcriptome analysis in breast cancer cells depleted of HP1γ and cross-referenced it with its genomic localisation, which identified increased expression of interferon/antiviral defense genes and activation of pro-apoptotic pathways. Whilst genes involved in these pathways were not directly bound by HP1γ, this analysis also identified HP1γ as a novel regulator of zinc finger (ZNF) genes. In summary, I identified novel post-translational modifications in HP1γ and characterised them in mESCs. I further demonstrated a role for HP1γ regulating breast cancer cell viability and identified HP1γ as a novel regulator of ZNF genes. My findings highlight HP1γ as a potential target for breast cancer therapy.

Made available in DSpace on 2014-06-11T19:22:31Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-20Bitstream added on 2014-06-13T18:07:11Z : No. of bitstreams: 1 santos_ea_me_ilha.pdf: 1369014 bytes, checksum: 7ea566c05320b595e08fe89ae2170203 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O teste de força de aperto das mãos tem como finalidade detectar eventuais patologias nos membros superiores e avaliar a força exercida pelas mãos de pacientes. Neste trabalho é descrito o desenvolvimento de um dinamômetro biomédico projetado para efetuar a avaliação funcional das mãos, através da utilização de moldes específicos. O equipamento é constituído basicamente por sensores com extensômetros metálicos, um circuito de condicionamento de sinais, um circuito de interfaceamento e por um display digital. O circuito de condicionamento de sinais foi implementado com um amplificador de instrumentação, um amplificador com ganho programável e por um filtro passa-baixas. O principal componente do circuito de interfaceamento é o microcontrolador ATMEGA8. O equipamento é robusto, apresenta resposta linear na faixa de 0 a 500 N, precisão de 0,54%, resolução de 0,70 N e histerese desprezível. O valor da força pode ser lida em um display e também na tela de um computador. Pode ser útil em Engenharia de Reabilitação, Fisioterapia e Terapia Ocupacional / In this work we describe the development of a biomedical dynamometer. It was designed with the goal of performing the functional evaluation of hands with specific shapes. The equipment is constituted by force sensors, a signal conditioning circuit, an interface circuit and a digital display. Metallic strain gages were used as sensors. The signal conditioning circuit is constituted by an instrumentation amplifier, a programmable gain amplifier and a low-pass filter. The main component of the interface circuit is an ATMEGA8 microcontroller. The instrument can measure forces with resolution of 0.70 N and precision of 0.54% in the range of 0 to 500 N. It is rugged, presents linear response and very small hysteresis. The value of the force can be read in a display and in a computer screen. The device can be useful in Rehabilitation Engineering, Physiotherapy and Ocupational Therapy

Orientadores: Antonio de Azevedo Barros Filho, Gil Guerra Junior / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-13T16:28:40Z (GMT). No. of bitstreams: 1 Ribeiro_RobertoRegis_D.pdf: 4470606 bytes, checksum: bb591117af61e1986e373f28d6447638 (MD5) Previous issue date: 2009 / Resumo: Estruturação da tese: O presente estudo optou pelo "Modelo Escandinavo", o qual é chamado de "Modelo Alternativo" no Programa de Pós-Graduação em Saúde da Criança e do Adolescente da Faculdade de Ciências Médicas da Universidade Estadual de Campinas (UNICAMP). Sendo assim, a tese foi composta por introdução, objetivos, capítulos (I e II) e conclusão geral. A introdução apresentou os principais estudos sobre massa óssea e da técnica de avaliação deste tecido por meio da ultrassonografia de falange. No capítulo I e II encontram-se os artigos que foram redigidos segundo as normas específicas de cada periódico a que foram submetidos. Na conclusão geral foram apresentadas as principais conclusões dos dois artigos. As referências bibliográficas foram apresentadas no final de cada capítulo e da tese. A seguir, foram apresentadas as principais informações que se referem à estruturação dos dois capítulos da tese. Objetivo: avaliar a quantidade e qualidade óssea, por meio de ultrassonografia das falanges, em escolares saudáveis, brancos e negros, em relação ao nível socioeconômico e composição corporal, e ainda comparar a quantidade óssea com referências européias. Materiais e Métodos: participaram do estudo 1.356 escolares saudáveis de diferentes níveis socioeconômicos, divididos em brancos e negros com idades de seis a onze anos, de ambos os gêneros. Por métodos antropométricos foram avaliados o peso e a estatura e calculados o percentual de gordura, massa gorda, massa magra e índice de massa corporal, que foi transformado em escore z para idade. Para definir os desvios de normalidade foram adotados como pontos de corte valores abaixo de -2 e acima de +2 dos valores do escore z. Para o procedimento de coleta das medidas individuais do parâmetro de quantidade óssea (AD-SoS = Amplitude Dependent of Speed Sound) e qualidade óssea (UBPI = Ultrasound Bone Profile Index) utilizou-se o ultra-som nas falanges com o aparelho DBM Sonic® BP, IGEA, de 3ª geração. Os valores médios de AD-SoS deste estudo foram comparados a cinco referências européias que apresentam valores normativos para crianças e adolescentes. Foi realizada a análise descritiva dos dados, e para a comparação da idade, dos gêneros e das cores da pele com as variáveis AD-SoS e UBPI, utilizou-se a análise de variância (ANOVA - two way) seguida pelo teste de comparações múltiplas de Tukey (p<0,05). Aplicaram-se ainda os testes Mann-Whitney, qui-quadrado, coeficiente de correlação e análise de regressão linear múltipla com significância de 5%. Para a comparação da AD-SoS dos escolares, de ambos os gêneros e cores de pele em relação às referências, recorreu-se ao teste t Student, que compara duas médias, sendo uma delas a referência. Resultados: Os escolares negros predominaram nos níveis socioeconômicos baixos. Os meninos negros apresentaram valores superiores no peso e estatura, enquanto as meninas negras na massa magra, em relação aos escolares brancos do mesmo gênero e idade. Observou-se variação crescente dos seis aos onze anos para ambos os gêneros e cor da pele nas médias de AD-SoS. Os brancos apresentaram valores superiores de AD-SoS e UBPI em relação aos negros. As variáveis antropométricas, gênero e nível socioeconômico explicaram apenas 17 e 11% da variabilidade de AD-SoS e UBPI, respectivamente. Comparando a AD-SoS deste estudo com as cinco referências européias, observou-se para ambos os gêneros e cores de pele valores inferiores ao estudo polonês e semelhante aos demais para as meninas, enquanto que os meninos apresentaram valores semelhantes aos estudos italianos e inferiores ao espanhol. Conclusão: Sendo assim, o presente estudo, apresentou massa óssea, avaliada por ultrassonografia das falanges, maior nos brancos que nos negros. A quantidade óssea dos escolares deste estudo aumentou dos seis aos onze anos de idade para ambos os gêneros e cores da pele. A quantidade óssea encontrada nas crianças brasileiras de ambos os gêneros e cores de pele foi inferior em relação aos poloneses, semelhante aos italianos; e em relação ao estudo espanhol, os gêneros não apresentaram uniformidades, sendo as meninas semelhantes e os meninos com valores inferiores. / Abstract: Thesis Structuring: The present study opted for the "Scandinavian Model" which is called the "Alternative Model" in the Post-Graduation Program in Child and Adolescent Health from the Medical Sciences College of University of Campinas (UNICAMP). Therefore, the thesis was compounded by introduction, objective, chapters (I and II) and general conclusion. The introduction presented the main studies on bone mass and the evaluation technique of this tissue by the use of phalanx ultrasound. The articles that were written according to the specific norms of each journal to which they were submitted to are found on chapters I and II. On the general conclusion the main conclusions of both articles were presented. The bibliographical references were presented at the end of each chapter and of the thesis. Further, the main information about the structuring of both chapters of the thesis was presented. Objective: to evaluate the bone quantity and quality, by phalanxes ultrasound, in healthy, black and white, schoolchildren, in relation to the socioeconomic level and body composition, and still, to compare the bone quantity to European references. Materials and Methods: 1,356 healthy schoolchildren from different socioeconomic level participated on this study, divided into black and white with ages from six to eleven years old, from both genders. The weight and height were evaluated by anthropometric methods as well as the percentage of fat, fat mass, lean mass and body mass index, which was transformed in score z for the age, were calculated. To define the normality deviations it was adopted, as cut points, values below -2 and above +2 from the values of the score z. For the procedure of collecting the individual data of the bone quantity parameter (AD-SoS = Amplitude Dependent of Speed Sound) and bone quality (UBPI = Ultrasound Bone Profile Index) it was used the ultrasound on the phalanxes with the 3rd generation DBM Sonic® BP, IGEA device. The average values of AD-SoS from this study were compared to five European references that presented normative values for children as well as for adolescents. The descriptive analysis of the data was realized, and for the age, genders and skin colors comparison with the variables AD-SoS and UBPI, the variance analysis was used (ANOVA - two way) followed by the test of multiple comparisons of Tukey (p<0,05). The Mann-Whitney, qui-squared, correlation coefficient and multiple linear regression analysis with 5% significance were also applied. For the comparison of AD-SoS of the schoolchildren, from both genders and skin colors in relation to the references, the t Student, that compares two averages, being one of them the reference, was chosen. Results: the black schoolchildren predominated on the low socio-economic levels. The black boys presented higher values for weight and height, while the black girls for lean mass, in relation to white schoolchildren from the same age and gender. It was observed a crescent variation from six to eleven years of age for both genders and color of skin in the averages of AD-SoS. The white schoolchildren presented higher values of AD-SoS and UBPI in relation to the black ones. The anthropometric variables, gender and socio-economic level explained only 17 and 11% of the variability of AD-SoS and UBPI, respectively. Comparing the AD-SoS from this study to the five European references, it was observed for both genders and colors of skin lower values than to the Polish study and similar to the other studies for the girls, while the boys presented similar values to the Italian studies and lower than the Spanish ones. This way, the present study, presented higher bone mass, evaluated by phalanxes ultrasound, in white schoolchildren than in black ones. The bone quantity of the schoolchildren from this study rose from the six to the eleven years of age for both genders and colors of skin. The bone quantity found in Brazilian children from both genders and colors of skin was lower in relation to the Polish, similar to the Italians; and in relation to the Spanish study, the genders didn't present uniformities, being the girls similar and the boys with lower values. / Doutorado / Saude da Criança e do Adolescente / Doutor em Saude da Criança e do Adolescente

Transcription is a major step in expression of genes of a given organism. Due to environmental constrains this step must be regulated in the favour of the sustenance and growth of the organism. Here comes the relevance of transcription factors, mostly proteins which regulate transcription. One such important group of transcription factors is the zinc finger proteins. It is well known that in eukaryotes the C2H2 zinc finger domain containing proteins are the largest group of transcription factors while in prokaryotes the largest group of transcription factors are represented by helix-turn-helix motif containing proteins. Till now only two C2H2 zinc finger domain proteins-Ros and Muc have been found in alpha proteobacteria which are also transcription factors. In eukaryotes the second largest group of zinc finger proteins have their zinc ion coordinated by four cysteine residues- the C4 zinc finger proteins. They make the nuclear hormone receptor superfamily of proteins. They have also been shown to act as transcription factors. But in eubacteria no such proteins have been described in details except an isolated report of crystal structure of a C-terminal zinc finger domain protein- Jann_2411 from Jannaschia sp. Though a lot of transcription factors have been described in mechanistic details in Escherichia coli and Bacillus subtilis, the list of well described mycobacterial transcription factors is short. Given this fact and the lack of any known zinc finger domain transcription factor in actinobacteria we wanted to see whether M. smegmatis genome also encode any homologue of Jann_2411 and if does whether they have ability to modulate transcription. To meet our aim we did BLASTP search against the genome of M. smegmatis using Jann_2411 as query. We found four C-terminal zinc finger domain proteins –Msmeg_0118. Msmeg_3613, Msmeg_3408 and Msmeg_1531, which we named as Mycobacterial single zinc finger protein (Mszfp) and numbered- Mszfp1, Mszfp2, Mszfp3 and Mszfp4, respectively. Mszfp1 and Mszfp2 were chosen for study as they were the top most hits. In this thesis:- Chapter1 introduces zinc finger proteins, transcription and several levels of control of transcription process in eubacteria. In chapter2 we characterised Mszfp1 biophysically and probed its secondary structure content and oligomeric state in the native and demetallated conditions. We have also shown that this conserved hypothetical protein is expressed throughout the growth phase of M. smegmatis, regulated by SigA and SigB. We have also showed that Mszfp1 is a DNA binding protein in the native state and the demetallated protein has altered DNA binding ability. It was noted that on over expression Mszfp1 affects colony morphology and biofilm forming ability, of M. smegmatis. In chapter3 the ability of Mszfp1 to bind to RNA polymerase of M. smegmatis has been explored. It was found that Mszfp1 can activate transcription by interacting with CTD/NTD of α subunit and domain 4 of σA like CRP on type II CRP activated promoter. In chapter4 similar to Mszfp1 the biophysical study of Mszfp2 has been carried out. It was found that Mszfp2 is also a predominantly alpha helical protein with oligomeric structure having DNA binding ability. Similar to Mszfp1 Mszfp2 on over expression changes the colony morphology. Chapter5 deals with the RNA polymerase binding ability of Mszfp2 and its ability to activate transcription by interacting with CTD/NTD of α subunit but not the σA. In chapter6 we have presented a glimpse of the possible biophysical properties of Mszfp3 and Mszfp4 and given a snapshot of distribution of homologues of Mszfps among other actinobacteria. We have also put forward a hypothesis about the origin of C4 and CCHC zinc finger domains. Chapter7 is the summary of the work embedded in the earlier chapters. In Appendix I is described the making of a bacteria (Bacillus licheniformis) driven heat engine. Appendix II describes an effort to study the visco-elastic properties of Mycobacterium smegmatis cells.

Previous publications from this laboratory demonstrated that administration of leukemia inhibitory factor (LIF) (125 µg/kg) to young, male Sprague-Dawley rats at 6, 24, and 48 h after middle cerebral artery occlusion (MCAO) reduced infract volume, improved sensimotor skills, and alleviated damage to white matter at 72 h after the injury. In vitro studies using cultured oligodendrocytes (OLs) showed that LIF (200 ng/ml) also protects against 24 h of oxygen-glucose deprivation through activation of Akt signaling and upregulation of the antioxidant enzymes peroxiredoxin IV and metallothionein III. Other groups have demonstrated that LIF reduces neurodegeneration in animal models of disease, but the neuroprotective mechanisms of LIF during permanent ischemia have not yet been examined. The overall hypothesis to be tested in this project is whether LIF exerts similar protective mechanisms against neurons during ischemia through increased antioxidant enzyme expression in neurons. In the first set of experiments, superoxide dismutase (SOD) activity was significantly increased in the ipsilateral hemisphere of LIF-treated rats compared to rats that received PBS treatment at 72 h after MCAO. Western blot and immunohistochemical analysis revealed that SOD3 was upregulated in brain tissue and induced specifically in cortical neurons tissue at this time point. Neurons that expressed high levels of SOD3 at 72 h after MCAO also showed high levels of phosphorylated Akt (Ser473). LIF (200 ng/ml) reduced necrotic and apoptotic cell death against 24 h of OGD as measured by lactate dehydrogenase (LDH) release and caspase-3 activation. Quantitative real-time PCR analysis showed that LIF treatment upregulated SOD3 gene expression in vitro during OGD. Treatment with 10 µM Akt Inhibitor IV and transfection with SOD3 siRNA counteracted the neuroprotective effects of LIF in vitro, showing that upregulation of SOD3 and activation of Akt signaling are necessary for LIF-mediated neuroprotection. Several transcription factors that regulated Akt-inducible genes were previously identified by this lab, including myeloid zinc finger-1 (MZF-1) and specificity protein-1 (Sp1). The goal of the second set of experiments was to determine whether LIF exerted protective actions through MZF-1 and Sp1. According to analysis with Genomatix, MZF-1 and Sp1 have multiple binding sites in the promoter for the rat SOD3 gene. Western blot analysis showed that there was a trend towards increased MZF-1 protein expression in the brains of LIF-treated rats that approached significance. Immunohistochemical analysis and quantitative real-time PCR showed a significant in vitro upregulation in MZF-1 expression among LIF-treated neurons compared to PBS-treated neurons. Sp1 gene expression was not changed by LIF treatment, but there was a trend towards increased protein expression. In addition, there was a significant correlation between Sp1 and MZF-1 among brain samples from LIF-treated rats but not PBS-treated or sham rats at 72 h after MCAO. Immunohistochemical analysis revealed that Sp1 and MZF-1 co-localized with neuronal nuclei and SOD3 at 72 h after MCAO. Neurons that were transfected with MZF-1 or Sp1 siRNA following isolation did not show a significant decrease in LDH release after 24 h OGD that was observed among neurons transfected with scrambled siRNA. These data demonstrate that Sp1 and MZF-1 are involved with the neuroprotective signaling of LIF under ischemia. This laboratory has demonstrated that LIF activates transcription of protective genes and increases the activity of transcription factors through modulation of intracellular signaling. However, the upstream signaling mechanisms of LIF during ischemia had not previously been investigated. Previous investigators found that the LIF-specific subunit of the heterodimeric LIF receptor (LIFR) is induced by CNS injury. Western blot analysis was used to determine whether LIFR was induced in the brain and the spleen, which plays a role in the peripheral immune response, after MCAO. According to these results, LIF treatment significantly upregulates LIF in the brain compared to PBS treatment or sham injury at 72 h after MCAO. Genomatix analysis of the LIFR promoter region revealed a binding site for Sp1, which is one of the transcription factors responsible for neuroprotection by LIF. At this same time point, splenic LIFR expression is significantly reduced after MCAO compared to sham injury. LIF treatment did not significantly increase LIFR expression, but did significantly increase spleen size compared to PBS treatment at 72 h after MCAO. Although there was a trend towards increased LIFR expression in the spleen from 24 h to 72 h after MCAO, this increase was not statistically significant. However, there was a significant positive correlation between spleen weight and LIFR expression among rats euthanized 24-72 h after MCAO/sham injury. In addition, there was a significant negative correlation between LIFR expression in the brain and the spleen weight, thus showing that LIFR is upregulated following the splenic response. According to findings from other groups, JAK1 has been shown to associate with the heterodimeric LIF receptor (LIFR/gp130) and directly activate PI3K/Akt signaling. To test whether JAK1 contributes neuroprotection during ischemia, cultured neurons were treated with several concentrations (2.5-50 nM) of GLPG0634, a JAK1-specific inhibitor prior to 24 h of OGD. With the exception of the 2.5 nM concentration, all concentrations of GLPG0634 significantly decreased LDH release compared to DMSO treatment, with the 5 nM concentration having the most potent effect on reducing cytotoxicity. However, the 5 nM concentration had no significant did not significantly reduce LDH release compared to DMSO treatment under 24 h of normoxic conditions. These results indicate that JAK1 activity is primarily detrimental to neurons during ischemia. Although it is possible that LIF signaling activates JAK1, it is unlikely that JAK1 is responsible for LIF-mediated neuroprotection during ischemia. The results of these experiments allowed us to determine several molecular mechanisms for LIF-mediated neuroprotection. LIF, which binds to its heterodimeric receptor, activates Akt signaling during ischemia. The transcription factors Sp1 and MZF-1, which are located downstream of Akt, bind to the promoter of the SOD3 gene. In addition, Sp1 also regulates the LIFR gene. SOD3 upregulation increases total SOD activity, which decreases apoptotic and necrotic cell death during apoptosis. Due to its ability to promote antioxidant expression and survival signaling in multiple neural cell types, LIF shows promise as a novel treatment for permanent focal cerebral ischemia.

1 Abstract Fingerprinting - Past, Present and Future The objective of the presented diploma thesis is to provide a comprehensive overview of findings in fingerprints area. Fingerprinting is a unique method enabling quick, inexpensive and reliable personal identification. To provide a comprehensive understanding of the presented issue, fingerprint reputation as ever-changing and continuously developing method is put into contrast with its contradictions and current problems. The first part of the thesis describes roots and genesis of the method from its beginnings in ancient realms up to date. The process of shaping of the knowledge in the field of fingerprints into a science is presented through the works of the most prominent fingerprints personalities. Despite the difficult beginnings, fingerprints finally reached the position of trustworthy and respectful area of forensic science. Due to that; the historical part follows the journey of fingerprints evidence into the police investigation and court rooms. Final point brings an attention to the establishment of fingerprinting in Czech criminal investigation. The main part of the thesis is dedicated to the practical aspect of fingerprints expertise. To provide a deep level of understanding, particular role of fingerprints in criminal investigations and its...