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1.
Folia Morphol (Warsz) ; 76(1): 100-109, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-27830891

RESUMO

BACKGROUND: Detailed study of the craniovertebral junction (CVJ) is necessary to completely understand the mechanism of its flexion and extension. MATERIALS AND METHODS: One cadaver head was sectioned in the sagittal plane. Also, in 22 volunteers, examined using the multislice computed tomography (MSCT), 14 parameters and 2 angles were measured in the neutral position, flexion and extension. RESULTS: The obtained measurements showed the anterior part of the occiput to move inferiorly in flexion, and the anterior atlas arch and the tip of the dens to get closer to the basion. At the same time, the opisthion moves superiorly, but the cervical spine bends anteriorly. Consequently, the dens-opisthion diameter and the opisthion-posterior atlas arch distance slightly decrease in length, whilst the arches of the atlas (C1), axis (C2) and C3 vertebra become more distant. Following extension, the posterior part of the occiput moves inferiorly, so that the basion-dens tip, the basion-axis arch, and the basion-posterior atlas arch distances increase in length. In contrast, the distances of the C1-C3 arches decrease in length. The angle between the foramen magnum and the dens tip decreases 1.620 on average in flexion, but increases 3.230 on average in extension. The angle between the axis body and the opisthion also decreases in flexion (mean, 3.360) and increases in extension (mean, 6.570). Among the congenital anomalies, a partial agenesis of the posterior atlas arch was revealed (4.5%), as well as an anterior dehiscence of the C1 foramen transversarium (13.6%). CONCLUSIONS: The mentioned measurements improved our understanding of the CVJ biomechanics. The obtained data can be useful in the evaluation of the CVJ instability caused by trauma, congenital anomalies and certain spine diseases.


Assuntos
Vértebra Cervical Áxis/diagnóstico por imagem , Atlas Cervical/diagnóstico por imagem , Tomografia Computadorizada Multidetectores , Adulto , Feminino , Humanos , Masculino
2.
Folia Morphol (Warsz) ; 74(4): 421-7, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26620500

RESUMO

BACKGROUND: The hippocampal formation (HF) is one of the most important parts of the brain in the magnetic resonance imaging (MRI) volumetric analysis in various domains, but not completely from all aspects, including the handedness. The aim of our study was to evaluate the possible differences in the volume of the right and left HF among the healthy right-handed and left-handed subjects, and to determine whether the volume differences are age related. MATERIALS AND METHODS: The MRI of this prospective study was performed using T1 fast field echo (FFE) sequence. The 124 subsequent coronal slices (thickness 1.5 mm) were performed in each participant. The obtained HF volumes were normalised and statistically compared. Volunteers comprised 30 persons aged 22.0 years, 12 of whom were the left-handed, and 30 persons aged 75.2 years on average, 9 of whom were the left-handed. RESULTS: The right and left HF volumes averaged 2.986 cm3 and 2.858 cm3 in the right-handed, and 2.879 cm³ and 3.020 cm³ in the left-handed young volunteers, as well as 2.728 cm³ and 2.650 cm³ in the right-handed, and 2.617 cm³ and 2.780 cm³ in the left-handed elderly persons. The HF volume ratios in the young left-handed participants showed a significant left-greater-than-right asymmetry. A significant difference was also noticed within the right-to-left volume ratios of the right- and left-handed young and elderly participants. The latter reduction in the HF volume within the aged group can be interpreted as a slight atrophy of the HF. CONCLUSIONS: There is a significant difference in the volumes of the left and right HF of the left-handed young participants. The age related HF volume differences were proven between the groups of the young and elderly volunteers. The obtained data should be included into the future MRI studies of the HF volumes in various clinical domains.

3.
Folia Morphol (Warsz) ; 72(3): 210-6, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-24068682

RESUMO

BACKGROUND: Data about the structure and immunohistochemistry of the lenticulostriate arteries (LSAs), although very important for medical research and clinical practice, have been rarely reported in literature. MATERIALS AND METHODS: Fourty serially sectioned LSAs were stained with hematoxilin and eosin, and prepared for immunohistochemistry. RESULTS: Our examination revealed a typical endothelial lining and a narrow subendothelial space with subintimal smooth muscle cells occasionally. The internal elastic lamina was fragmented or absent in the smallest LSAs branches. The mediacoat, with a mean diameter of 148.5 µm, contained typical smooth muscle cells which formed 14.2 layers on average and showed a positive immune reactions for alfa-actin, desmine, laminin and collagen IV. The thin adventitial coat contained fibroblasts, collagen fibers, and nerve bundles, with the strongest immunopositivity to thyrosin hydroxilase. The immune reactions against CD31 and CD34 proteins,endothelial nitric oxide synthase, S 100 protein, neurofilament protein and synaptophysin,seem to be performed in the LSAs wall for the first time. Similarly,the thickness of the LSAs wall and its coats have never been reported, nor the number of the smooth muscle cell layers. CONCLUSIONS: Our results related to the structure and immunohistochemistry of the LSAs could be important in cerebrovascular pathology, neurology and neurosurgery.


Assuntos
Artéria Cerebral Média/anatomia & histologia , Artéria Cerebral Média/metabolismo , Proteínas/metabolismo , Adolescente , Adulto , Túnica Adventícia/anatomia & histologia , Túnica Adventícia/metabolismo , Idoso , Feminino , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Túnica Íntima/anatomia & histologia , Túnica Íntima/metabolismo , Túnica Média/anatomia & histologia , Túnica Média/metabolismo , Adulto Jovem
4.
Folia Morphol (Warsz) ; 72(2): 113-22, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-23740497

RESUMO

BACKGROUND: The most reliable data about arterial variations, which are very important in surgery and radiology, can be obtained from a large series of patients. MATERIALS AND METHODS: We examined angiographic and multislice computerised tomography (MSCT) images in a group of 1,265 patients and in 1 dissected specimen. RESULTS: While in 946 (74.72%) of the patients a normal vascular pattern (type I) was noticed, in the remaining 320 (25.28%) patients variations of the branches of the aortic arch were found, which were classified into types II through VIII and a few subtypes. Type II (2.84%) comprised a common origin of the left commoncarotid and subclavian arteries. Type III (15.56%) was related to an origin of the left subclavian artery from the brachiocephalic trunk. Type IV (0.55%) included the aortic origin of both common carotid and subclavian arteries, with the right subclavian artery having a retroesophageal course. Type V (0.24%) included the same 4 supra-aortic branches, which, however, arose from a double or a right--sided aortic arch. Type VI (3.63%) comprised the aortic origin of the left vertebral artery, type VII (0.24%) the same origin of the right vertebral artery, and type VIII(2.22%) the aortic origin of the thyroideaima artery. A corresponding embryological background and clinical implications of the described aberrant vessels were presented. CONCLUSIONS: In more than one quarter of the cases, the branching pattern of the examined arteries did not follow the classical pattern. Detailed knowledge of aortic branch variations is of great significance in anatomy, embryology, and clinical medicine, especially in radiology and thoracic surgery.


Assuntos
Aorta Torácica/anatomia & histologia , Aorta Torácica/diagnóstico por imagem , Idoso , Angiografia/métodos , Feminino , Humanos , Masculino
5.
Pharmazie ; 66(9): 662-5, 2011 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22026120

RESUMO

Cellular drug target identification through affinity chromatography is often hindered by the quantity of nonspecific binders, such as cytoskeletal and heat shock proteins. Thus, we prepared a 2-aminobenzimidazole-tethered depletion resin designed for removal of these proteins, and tested it on human lung carcinoma cell and rat tissue extracts. Column-bound proteins were identified by two-dimensional gel electrophoresis and MS. Among others, tubulins, actins and heat shock proteins were successfully depleted. Due to the reduction of these highly abundant proteins detection of potential drug targets is considerably facilitated in the pre-purified samples.


Assuntos
Benzimidazóis/química , Células/química , Proteínas/isolamento & purificação , Extratos de Tecidos/química , Marcadores de Afinidade , Animais , Química Encefálica , Carbono/análise , Linhagem Celular Tumoral , Cromatografia Líquida de Alta Pressão , Ecocardiografia , Humanos , Hidrólise , Indicadores e Reagentes , Masculino , Ratos , Ratos Wistar , Propriedades de Superfície , Espectrometria de Massas em Tandem , Tripsina/química
6.
Acta Biol Hung ; 62(4): 413-25, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22119870

RESUMO

Micro RNAs (miRNA) are an abundant class of small RNAs that regulate the stability and translation of cognate mRNAs. MiRNAs are potential diagnostic markers, moreover, they play an essential role in the development of various heart disesases. In case of limited tissue material, such as, e.g. human biopsies, purification of miRNAs with sufficient yield is critical. Reproducible expression analysis of miRNAs is highly dependent on the quality of the RNA, which is often difficult to achieve from fibrous tissue such as the heart. Several companies developed general purification kits for miRNAs, however, none of them are specialized to fibrotic tissues. Here we describe an optimized miRNA purification protocol that results in high miRNA yield as compared to other methods including trizol-based and column-based protocols. By using our improved protocol, miRNA obtained from heart tissue gave more reproducible results in QRT-PCR analysis and obtained more significant calls (172 vs. 118) during DNA microarray analysis when compared to the commercially available kit. In addition to the heart tissue, the present protocol can be applied to other fibrotic tissues, such as lung or skeletal muscle to isolate high-purity miRNA.


Assuntos
MicroRNAs/química , Miocárdio/química , Animais , Camundongos , Análise de Sequência com Séries de Oligonucleotídeos , Ratos , Reação em Cadeia da Polimerase em Tempo Real
7.
J BUON ; 16(4): 727-32, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-22331729

RESUMO

PURPOSE: It is known that expression disorders of cell cycle regulators play an important role in the development and prognosis of various malignant tumors. Cyclin expression changes during the cell cycle. This work aimed to analyse the expression of cyclin E in transitional cell carcinoma (TCC) and also to compare the expression of cyclin E with tumor stage and histological grade as well as to determine possible existence of differences in the expression of cyclin E in TCCs of the upper and lower urothelium. METHODS: Twenty-four cases of TCC of the urinary tract were retrospectively analysed (6 cancers of the renal pelvis, 2 of the ureter and 15 of the bladder; 4 were infiltrative). Immunohistochemical staining for cyclin E of the analysed transitional cancer cells was assessed semiquantitatively: diffuse cyclin E expression + + + (> 50% of all cells), expression in larger groups of cells: + + (up to 50% of all cells), expression in individual cells or small cell clusters: + (<10% of all cells), and absence of expression. Tumor stage was based on clinical and morphological criteria. WHO classification (Lyon 2004) was used for determination of the histological grade. RESULTS: Non-parametric Spearman's correlation showed that there was no statistically significant correlation between tumor stage and expression of cyclin E (ρ = -0331, p> 0.05). Also, no statistically significant correlation between grade and the expression of cyclin E (ρ = -0077, p> 0.05) was found. x2 test results showed no statistically significant difference (x2 = 2.136, p = 0.775) in the expression of cyclin E between upper and lower urothelium. CONCLUSION: This study showed non significant decreased expression of cyclin E with poor differentiation, muscle invasion and upper/lower urothelium. Expression of cyclin E decreased with increasing histological grade and stage of the tumor.


Assuntos
Carcinoma de Células de Transição/metabolismo , Ciclina E/biossíntese , Neoplasias da Bexiga Urinária/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinoma de Células de Transição/patologia , Feminino , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Prognóstico , Estudos Retrospectivos , Neoplasias da Bexiga Urinária/patologia
8.
Acta Biol Hung ; 61(4): 434-48, 2010 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-21112835

RESUMO

The aim of the present study was to identify gene expression changes in the rapid cardiac pacing-induced delayed antiarrhythmic protection in the canine, using cDNA microarrays and quantitative real-time PCR (QRT -PCR) techniques. In all dogs under light pentobarbitone anaesthesia, a pacing electrode was introduced into the right ventricle, and then the animals were divided into three groups: (1) sham-operated and sham-paced group (SP, n = 3) (2) ischaemic control group (IC; n = 3); these were without cardiac pacing and subjected only to a 25 min occlusion of the left anterior descending coronary artery (LAD), and (3) paced group (PC, n = 3); these animals were paced at a rate of 220-240 beats min-1 24 h prior to ischaemia. With cDNA chip 23 genes were found with altered expression in response to rapid cardiac pacing and 10 genes in the IC group when compared to SP dogs. These genes encode transcription factors (MEF2); members of signaling pathways (TGFß2, PDE4D9), hormone related proteins (e.g. vasopressin V1 and V2 receptors). RT-QPCR was used either to confirm the results of the microarray analysis and also to study 46 genes which are already known to have a role in the late phase of PC. By this method 17 genes were up-regulated and 6 genes down-regulated in the IC group; their expression ratios changed either to the opposite or showed no alteration after cardiac pacing. This study would add some new information about those transcriptional changes that are involved in the delayed phase of cardiac protection.


Assuntos
Estimulação Cardíaca Artificial/métodos , Perfilação da Expressão Gênica , Pentobarbital/farmacologia , Anestesia , Animais , Vasos Coronários/patologia , DNA Complementar/metabolismo , Cães , Regulação para Baixo , Feminino , Coração/fisiologia , Masculino , Análise de Sequência com Séries de Oligonucleotídeos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Regulação para Cima
9.
Cell Death Differ ; 14(6): 1181-90, 2007 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-17363962

RESUMO

In Drosophila, the fat body undergoes a massive burst of autophagy at the end of larval development in preparation for the pupal transition. To identify genes involved in this process, we carried out a microarray analysis. We found that mRNA levels of the homologs of Atg8, the coat protein of early autophagic structures, and lysosomal hydrolases were upregulated, consistent with previous results. Genes encoding mitochondrial proteins and many chaperones were downregulated, including the inhibitor of eIF2alpha kinases and the peptidyl-prolyl cis-trans isomerase FK506-binding protein of 39 kDa (FKBP39). Genetic manipulation of FKBP39 expression had a significant effect on autophagy, potentially through modulation of the transcription factor Foxo. Accordingly, we found that Foxo mutants cannot properly undergo autophagy in response to starvation, and that overexpression of Foxo induces autophagy.


Assuntos
Autofagia/genética , Proteínas de Drosophila/genética , Drosophila/genética , Corpo Adiposo/metabolismo , Perfilação da Expressão Gênica , Proteínas de Ligação a Tacrolimo/genética , Animais , Autofagia/fisiologia , Drosophila/crescimento & desenvolvimento , Proteínas de Drosophila/fisiologia , Fatores de Transcrição Forkhead/genética , Fatores de Transcrição Forkhead/fisiologia , Regulação da Expressão Gênica no Desenvolvimento , Larva/genética , Mutação , Análise de Sequência com Séries de Oligonucleotídeos , PTEN Fosfo-Hidrolase/genética , PTEN Fosfo-Hidrolase/fisiologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Proteínas de Ligação a Tacrolimo/fisiologia
10.
RSC Adv ; 8(52): 29924-29927, 2018 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-35547277

RESUMO

The oxidation of Hantzsch ester by a pyrylium cation takes place via electron-proton-electron transfer. The reaction was investigated with EPR spectroscopy using TEMPO and DMPO for inhibition and spin trapping, respectively, of the radicals appearing during the reaction. The present in-depth EPR study of the radical reactions of a NADH analogue indicate a complex electron transfer mechanism in the title reaction.

11.
Biochim Biophys Acta ; 1759(11-12): 543-51, 2006.
Artigo em Inglês | MEDLINE | ID: mdl-17182124

RESUMO

Alfalfa leaf protoplast-derived cells can develop into somatic embryos depending on the concentration of 2,4-dichlorophenoxyacetic acid (2,4-D) in the initial culture medium. In order to reveal gene expression changes during the establishment of embryogenic competence, we compared the cell types developed in the presence of 1 and 10 microM 2,4-D, respectively, at the time of their first cell divisions (fourth day of culture) using a PCR-based cDNA subtraction approach. Although the subtraction efficiency was relatively low, applying an additional differential screening step allowed the identification of 38 10 microM 2,4-D up-regulated transcripts. The corresponding genes/proteins were annotated and representatives of various functional groups were selected for more detailed gene expression analysis. Real-time quantitative PCR (RT-QPCR) analysis was used to determine relative expression of the selected genes in 2,4-D-treated leaves as well as during the whole process of somatic embryogenesis. Gene expression patterns confirmed 2,4-D inducibility for all but one of the 11 investigated genes as well as for the positive control leafy cotyledon1 (MsLEC1) gene. The characterized genes exhibited differential expression patterns during the early induction phase and the late embryo differentiation phase of somatic embryogenesis. Genes coding for a GST-transferase, a PR10 pathogenesis-related protein, a cell division-related ribosomal (S3a) protein, an ARF-type small GTPase and the nucleosome assembly factor family SET protein exhibited higher relative expression not only during the induction of somatic embryogenesis but at the time of somatic embryo differentiation as well. This may indicate that the expression of these genes is associated with developmental transitions (differentiation as well as de-differentiation) during the process of somatic embryogenesis.


Assuntos
Genes de Plantas/genética , Medicago sativa/genética , Folhas de Planta/genética , Protoplastos/metabolismo , Ácido 2,4-Diclorofenoxiacético/farmacologia , DNA Complementar/química , DNA Complementar/genética , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Biblioteca Gênica , Medicago sativa/citologia , Medicago sativa/embriologia , Dados de Sequência Molecular , Folhas de Planta/citologia , Folhas de Planta/embriologia , Proteínas de Plantas/classificação , Proteínas de Plantas/genética , Proteínas de Plantas/fisiologia , Protoplastos/citologia , Análise de Sequência de DNA
12.
Neuroscience ; 314: 47-63, 2016 Feb 09.
Artigo em Inglês | MEDLINE | ID: mdl-26633263

RESUMO

Microglial activation results in profound morphological, functional and gene expression changes that affect the pro- and anti-inflammatory mechanisms of these cells. Although statins have beneficial effects on inflammation, they have not been thoroughly investigated for their ability to affect microglial functions. Therefore the effects of rosuvastatin, one of the most commonly prescribed drugs in cardiovascular therapy, either alone or in combination with bacterial lipopolysaccharide (LPS), were profiled in pure microglial cultures derived from the forebrains of 18-day-old rat embryos. To reveal the effects of rosuvastatin on a number of pro- and anti-inflammatory mechanisms, we performed morphometric, functional and gene expression studies relating to cell adhesion and proliferation, phagocytosis, pro- and anti-inflammatory cytokine (IL-1ß, tumor necrosis factor α (TNF-α) and IL-10, respectively) production, and the expression of various inflammation-related genes, including those related to the above morphological parameters and cellular functions. We found that microglia could be an important therapeutic target of rosuvastatin. In unchallenged (control) microglia, rosuvastatin inhibited proliferation and cell adhesion, but promoted microspike formation and elevated the expression of certain anti-inflammatory genes (Cxcl1, Ccl5, Mbl2), while phagocytosis or pro- and anti-inflammatory cytokine production were unaffected. Moreover, rosuvastatin markedly inhibited microglial activation in LPS-challenged cells by affecting both their morphology and functions as it inhibited LPS-elicited phagocytosis and inhibited pro-inflammatory cytokine (IL-1ß, TNF-α) production, concomitantly increasing the level of IL-10, an anti-inflammatory cytokine. Finally, rosuvastatin beneficially and differentially affected the expression of a number of inflammation-related genes in LPS-challenged cells by inhibiting numerous pro-inflammatory and stimulating several anti-inflammatory genes. Since the microglia could elicit pro-inflammatory responses leading to neurodegeneration, it is important to attenuate such mechanisms and promote anti-inflammatory properties, and develop prophylactic therapies. By beneficially regulating both pro- and anti-inflammatory microglial functions, rosuvastatin may be considered as a prophylactic agent in the prevention of inflammation-related neurological disorders.


Assuntos
Anti-Inflamatórios/farmacologia , Inflamação/metabolismo , Inflamação/fisiopatologia , Microglia/efeitos dos fármacos , Microglia/metabolismo , Rosuvastatina Cálcica/farmacologia , Animais , Adesão Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Citocinas/metabolismo , Expressão Gênica/efeitos dos fármacos , Inflamação/induzido quimicamente , Inflamação/genética , Mediadores da Inflamação/metabolismo , Lipopolissacarídeos , Microglia/citologia , Microglia/fisiologia , Fagocitose/efeitos dos fármacos , Prosencéfalo/citologia , Ratos , Ratos Sprague-Dawley
13.
Psychiatr Genet ; 15(1): 1-6, 2005 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-15722950

RESUMO

Since the function and metabolism of peripheral lymphocytes is known to be altered in Alzheimer's disease (AD), a pilot study was carried out to examine differences in gene expression profiles of these cells in 16 AD patients and aged control probands. Using a cDNA microarray representing 3200 distinct human genes, we identified 20 candidate genes whose expression is altered in AD lymphocytes compared with the control probands. Among these were the alpha2C-adrenoreceptor gene, known to regulate blood pressure and learning, the defensin, histocompability complex enhancer-binding protein, carboxypeptidase M, and the Fc fragment of IgE known to be involved in cellular and humoral immune responses. Others, like human cell death protein, TRAIL, and galectin-4 participate in the regulation of apoptosis. Real-time quantitative reverse transcription-polymerase chain reaction analysis was performed in order to confirm the expression changes in AD lymphocytes, and it could detect down-regulation of defensin and alpha2c-adrenoceptor genes, while other genes seemed unaltered in their expression, including heat-shock protein (hsp90), cholesteryl ester transfer protein, and apolipoprotein B100 (apoB). The altered expression profile of these genes might be connected with the previously reported AD-specific lymphocyte abnormalities. It remains to be elucidated, however, how these genes are related to the pathomechanism of dementia and whether the gene expression differences of AD lymphocytes reflect disease traits or stage processes.


Assuntos
Doença de Alzheimer/genética , Perfilação da Expressão Gênica , Linfócitos/fisiologia , Idoso , Apolipoproteína B-100 , Apolipoproteínas B/genética , Primers do DNA , Instituição de Longa Permanência para Idosos , Humanos , Proteínas do Tecido Nervoso/genética , Casas de Saúde , Análise de Sequência com Séries de Oligonucleotídeos , Reação em Cadeia da Polimerase
14.
Cell Mol Biol (Noisy-le-grand) ; 51(2): 177-86, 2005 Sep 05.
Artigo em Inglês | MEDLINE | ID: mdl-16171553

RESUMO

Thyroid nodules are common. It would very helpful if genetic markers that can diagnose malignancy from fine needle aspiration samples were available. Few such markers has been thus identified and none are specific. Large panels of potential markers can be screened by microarray technology. Studies done to date have concentrated on single tumor types and thus provide no help in identifying tumor subtype specific markers. To that end we have studied gene profiles of 5 types of benign and malignant thyroid nodular tissue (multinodular goiter, follicular adenoma, papillary and follicular carcinomas). We have identified 195 genes whose differential expression clustered into clinically relevant groups. Twenty-eight genes were selected for further confirmation using real time quantitative polymerase chain reaction. Despite the differences in the microarray panels used, we confirmed the differential regulation of 12 genes previously reported in thyroid cancer, although we found the expression of several genes to be regulated in other histological tumor subtypes than originally described. We found, PCSK2, TRIB1, RAP1 GA1 to be specifically overexpressed in follicular cancer and S100A4 and GK2 in papillary carcinoma. SERP1, RNASE 2 and STATA5 were suppressed in papillary thyroid cancer. We have thus identified new potential markers specific to malignant thyroid tumors. It is apparent that a range of nodular thyroid tissue using large tumor sample numbers is necessary to establish robust markers for malignancy and to categorize tumors on the basis of small tumor samples.


Assuntos
Perfilação da Expressão Gênica , Genes Neoplásicos , Neoplasias da Glândula Tireoide/diagnóstico , Neoplasias da Glândula Tireoide/genética , Adenocarcinoma Folicular/diagnóstico , Adenocarcinoma Folicular/genética , Adenocarcinoma Folicular/fisiopatologia , Adenoma/diagnóstico , Adenoma/genética , Adenoma/fisiopatologia , Biomarcadores Tumorais/genética , Biópsia por Agulha Fina , Carcinoma Papilar/diagnóstico , Carcinoma Papilar/genética , Carcinoma Papilar/fisiopatologia , Regulação Neoplásica da Expressão Gênica , Bócio Nodular/diagnóstico , Bócio Nodular/genética , Bócio Nodular/fisiopatologia , Humanos , Microscopia Confocal , Análise de Sequência com Séries de Oligonucleotídeos , Reação em Cadeia da Polimerase , Neoplasias da Glândula Tireoide/fisiopatologia
15.
Acta Biol Hung ; 56(3-4): 233-45, 2005.
Artigo em Inglês | MEDLINE | ID: mdl-16196199

RESUMO

Recent and historical evidence is consistent with the view that atherosclerosis is an infectious disease or microbial toxicosis impacted by genetics and behavior. Because small bacterial-like particles, also known as nanobacteria have been detected in kidney stones, kidney and liver cyst fluids, and can form a calcium apatite coat we posited that this agent is present in calcified human atherosclerotic plaques. Carotid and aortic atherosclerotic plaques and blood samples collected at autopsy were examined for nanobacteria-like structures by light microscopy (hematoxylin-eosin and a calcium-specific von Kossa staining), immuno-gold labeling for transmission electron microscopy (TEM) for specific nanobacterial antigens, and propagation from homogenized, filtered specimens in culture medium. Nanobacterial antigens were identified in situ by immuno-TEM in 9 of 14 plaque specimens, but none of the normal carotid or aortic tissue (5 specimens). Nanobacteria-like particles were propagated from 26 of 42 sclerotic aorta and carotid samples and were confirmed by dot immunoblot, light microscopy and TEM. [3H]L-aspartic acid was incorporated into high molecular weight compounds of demineralized particles. PCR amplification of 16S rDNA sequences from the particles was unsuccessful by traditional protocols. Identification of nanobacteria-like particles at the lesion supports, but does not by itself prove the hypothesis that these agents contribute to the pathogenesis of atherosclerosis, especially vascular calcifications.


Assuntos
Arteriosclerose/microbiologia , Arteriosclerose/patologia , Bactérias/patogenicidade , Calcinose , Adulto , Aorta/microbiologia , Aorta/patologia , Aorta/ultraestrutura , Ácido Aspártico/metabolismo , Artérias Carótidas/microbiologia , Artérias Carótidas/patologia , Artérias Carótidas/ultraestrutura , Humanos , Hidroxiapatitas/química , Imuno-Histoquímica , Tamanho da Partícula
16.
Biomed Res Int ; 2015: 968981, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26075279

RESUMO

Combination therapy of bortezomib with other chemotherapeutics is an emerging treatment strategy. Since both curcumin and bortezomib inhibit NF-κB, we tested the effects of their combination on leukemia cells. To improve potency, a novel Mannich-type curcumin derivative, C-150, was synthesized. Curcumin and its analogue showed potent antiproliferative and apoptotic effects on the human leukemia cell line, HL60, with different potency but similar additive properties with bortezomib. Additive antiproliferative effects were correlated well with LPS-induced NF-κB inhibition results. Gene expression data on cell cycle and apoptosis related genes, obtained by high-throughput QPCR, showed that curcumin and its analogue act through similar signaling pathways. In correlation with in vitro results similar additive effect could be obsereved in SCID mice inoculated systemically with HL60 cells. C-150 in a liposomal formulation given intravenously in combination with bortezomib was more efficient than either of the drugs alone. As our novel curcumin analogue exerted anticancer effects in leukemic cells at submicromolar concentration in vitro and at 3 mg/kg dose in vivo, which was potentiated by bortezomib, it holds a great promise as a future therapeutic agent in the treatment of leukemia alone or in combination.


Assuntos
Apoptose/efeitos dos fármacos , Bortezomib/farmacologia , Curcumina/farmacologia , Leucemia/tratamento farmacológico , Leucemia/metabolismo , Animais , Curcumina/análogos & derivados , Células HL-60 , Humanos , Leucemia/patologia , Camundongos , Camundongos SCID , Ensaios Antitumorais Modelo de Xenoenxerto
17.
Curr Med Chem ; 11(23): 3119-45, 2004 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-15579004

RESUMO

Chemical genomics, which utilizes specially designed small chemical compounds early in the discovery phase of new drugs to explore the life science at various levels, can address biological questions that are not amenable to genetic manipulation or functional genomics/proteomics approaches. Following the development of HT phenotypic assays and DNA expression analysis, the integration of cell-based assays with activity / affinity-based approaches allows us to interrogate the cells by analyzing phenotypic alterations, changes of transcript signature or detecting the differences in protein expression levels. Furthermore, activity / affinity-based techniques directly provide a druggable subset of gene products, which interact with small molecules, greatly reducing the complexity of analyzing the proteome. In this paper, we give an account of the recent advances (approaches and strategies) in the field of chemical genomics, and discuss how these approaches enable the investigator to obtain a novel therapeutically relevant target as well as drug candidates acting on them in a target-specific manner. This novel post-genomic discovery strategy, where target identification/ validation is carried out by interactions with small molecules, could significantly reduce the time-scale for early drug discovery, and increase the success rate of finding novel, druggable targets, as well as more specific drug candidates.


Assuntos
Engenharia Química/métodos , Engenharia Química/tendências , Genômica/métodos , Genômica/tendências , Animais , Regulação da Expressão Gênica/fisiologia , Humanos , Ligantes , Ligação Proteica/fisiologia
18.
Biotechniques ; 33(4): 898-900, 902, 904, 2002 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-12398199

RESUMO

In DNA microarray technology, repeatability and reliability are very important to compare multiple RNA samplesfrom different experiments. The application of common or universal RNA as a standard control equalizes the differences in hybridization parameters and array variations. For this purpose, high-quality reference RNA is necessary in bulk amounts. A novel approach was developed to get milligrams of sense or antisense RNA, starting from micrograms of pooled total RNA from different cell lines, tissues, or organisms. This method is inexpensive and allows further labeling procedures using poly(dT) or random oligomers as primers. In addition, amplified, sense reference RNA is suitable for standard labeling protocols, while the antisense reference RNA can be used with antisense RNA from the linear sample amplification method. Here we produced universal RNA for human, rat, and alfalfa and demonstrated the quality using specific cDNA microarrays.


Assuntos
Análise de Sequência com Séries de Oligonucleotídeos/métodos , RNA/isolamento & purificação , Animais , Sequência de Bases , Humanos , Hibridização de Ácido Nucleico , Reação em Cadeia da Polimerase , Ratos
19.
Neurosurgery ; 28(4): 523-9; discussion 529-30, 1991 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-2034346

RESUMO

The thalamogeniculate (TG) arteries of 30 forebrain hemispheres were examined. These vessels varied from 2 to 12 in number (mean, 5.7), and from 70 to 580 microns in caliber (mean, 345.8 microns). The average caliber of all the TG vessels per posterior cerebral artery ranged from 700 to 3400 microns (mean, 1972 microns). The TG arteries most often originated as individual vessels; however, in 26.67% of the hemispheres examined they shared a common site of origin, and 33.33% of the hemispheres they arose from common stems. The common stems ranged from 320 to 800 microns in diameter (mean, 583 microns). The TG branches arose from the crural or ambient (P2) segment of the posterior cerebral artery in 80% of the hemispheres, from the P2 and the quadrigeminal (P3) segment in 20%, from both the distal segment of the posterior cerebral artery and the common temporal artery (13.33%), or from the distal segment and either the calcarine (3.33%) or parieto-occipital artery (3.33%). The TG arteries usually penetrated the medial geniculate body (100%), pulvinar thalami (80%), brachium of the superior colliculus (53.33%), or lateral geniculate body (13.33%). The collateral branches of the TG arteries were noted to reach the medial geniculate body (76.67%), pulvinar (70%), brachium of the superior colliculus (40%), crus cerebri (40%), and lateral geniculate body (6.67%). The anastomoses were present in 66.67%, usually between the TG vessels and the medial posterior choroidal artery (33.33%), or the mesencephalothalamic artery (26.67%). They ranged in number from 1 to 3 (mean, 1.2), and in caliber from 90 to 400 microns (mean, 197 microns).(ABSTRACT TRUNCATED AT 250 WORDS)


Assuntos
Artérias Cerebrais/anatomia & histologia , Corpos Geniculados/irrigação sanguínea , Tálamo/irrigação sanguínea , Artérias Cerebrais/cirurgia , Humanos , Microcirurgia , Valores de Referência , Colículos Superiores/irrigação sanguínea
20.
Surg Neurol ; 37(5): 339-49, 1992 May.
Artigo em Inglês | MEDLINE | ID: mdl-1631758

RESUMO

The hippocampal vessels were examined in 25 forebrain hemispheres injected with india ink or methylmethacrylate. There were two to seven hippocampal arteries, which measured 200-800 microns in diameter. The anterior hippocampal artery (AHA), which was present in 88.2% of the hemispheres, most often originated from the posterior cerebral and the anterior temporal arteries, that is, within the rostral hippocampo-parahippocampal arterial complex. It arose from the anterior choroidal artery in 29.4% of the hemispheres. The AHA extended between the uncus and the parahippocampal gyrus, and it supplied the head of the hippocampus. The middle hippocampal artery was constant. It most often arose from the posterior cerebral and the common temporal arteries. The middle hippocampal artery coursed just caudal to the uncus, in close relationship with the lateral posterior choroidal artery, and it usually supplied the middle part of the hippocampal formation. The posterior hippocampal artery, which existed in 94.1% of the hemispheres, most often arose from the posterior cerebral and the splenial arteries. It irrigated the caudal part of the hippocampal formation. The anastomoses connecting the posterior, middle, and the anterior hippocampal arteries were present in 29.4% of the hemispheres. The hippocampal arteries gave rise to the straight vessels, which divided into the large and the small intrahippocampal arteries. The highest density of the capillary network was noted in the pyramidal and molecular layers of the hippocampal formation. The clinical significance of the obtained microanatomical findings is discussed.


Assuntos
Hipocampo/irrigação sanguínea , Artérias/anatomia & histologia , Anastomose Arteriovenosa/anatomia & histologia , Cadáver , Humanos , Microcirculação
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