RESUMO
Food allergies have increased in recent years in Japan. Details of causative foods, places where anaphylaxis developed, and other allergic factors remain unknown, and we investigated them. A'questionnaire survey for the prevention of food allergies' was conducted using a nationwide group of patients with food allergies. A total of 1383 patients from 878 families (including 319 patients who experienced anaphylaxis) provided valid answers to the questionnaire. The average age of the first anaphylactic attack was 3.20 +/- 6.327 yr. The most common allergens causing anaphylaxis were in order milk, eggs, wheat, peanuts, and soybeans, followed by sesame and buckwheat. The most common place where anaphylaxis developed was the patient's own home, followed by fast food restaurants, places visited, restaurants, and schools. In patients' own homes, fast food restaurants (buffet), places visited and schools, the most common allergens were milk, eggs, and wheat. In restaurants and accommodation facilities, eggs were the most common allergen followed by milk. As possible food allergies can cause anaphylaxis, it is necessary to provide precise information for consumers regarding packaged and processed foods.
Assuntos
Alérgenos , Anafilaxia/epidemiologia , Hipersensibilidade Alimentar/epidemiologia , Adolescente , Adulto , Fatores Etários , Criança , Pré-Escolar , Coleta de Dados , Feminino , Humanos , Lactente , Japão/epidemiologia , Masculino , Inquéritos e QuestionáriosRESUMO
We found a 66-year-old Japanese patient with type I congenital heparin cofactor (HC) II deficiency manifesting multiple atherosclerotic lesions. To investigate the molecular pathogenesis of our patient, we performed sequencing analysis and expressed recombinant human wild-type and mutant HC II molecules in COS-1 and CHO-K1 cells. Sequencing analysis following amplification of each of all 5 exons and its flanking region showed a single C to T transition at nucleotide position 12,854 in exon 5, which changed a Pro443 codon (CCG) to Leu codon (CTG). Because this mutation generates a new Bhv I site, the Bbv I digestion pattern of the PCR-amplified exon 5 fragments from each family member was analyzed. In all cases, the patterns were consistent with the activities and antigen levels of plasma HC I1 in those members. Transient transfection, metabolic labeling and pulse-chase experiments followed by immunoprecipitation analysis showed that the recombinant mutant HC II molecules were secreted from COS-1 cells in reduced amounts compared with the wild-type, and that an enhanced intracellular association of the mutant molecules with a chaperone, GRP78/BiP, was observed in CHO-K1 cells. Northern blot analysis indicated that the mutant HC I1 mRNA was transcribed at a similar level as that of wild-type. Immunohistochemical staining of the transfected cells revealed that COS-1 cells expressing the mutant HC II molecules were stained mainly in the perinuclear area. We conclude that the impaired secretion of the mutant HC II molecules, due to intracellular degradation, is the molecular pathogenesis of type I congenital HC II deficiency caused by a Pro443 to Leu mutation at reactive P2 site.
Assuntos
Sítios de Ligação/genética , Proteínas de Choque Térmico , Cofator II da Heparina/deficiência , Mutação de Sentido Incorreto , Idoso , Animais , Arteriosclerose/etiologia , Arteriosclerose/genética , Células COS , Proteínas de Transporte/metabolismo , Análise Mutacional de DNA , Chaperona BiP do Retículo Endoplasmático , Saúde da Família , Feminino , Cofator II da Heparina/genética , Cofator II da Heparina/metabolismo , Humanos , Masculino , Chaperonas Moleculares/metabolismo , Linhagem , Protrombina/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Inibidores de Serina Proteinase/deficiência , Inibidores de Serina Proteinase/genética , Inibidores de Serina Proteinase/metabolismo , TransfecçãoRESUMO
The proband, a 76-year-old woman, suffered from dural arteriovenous fistula. Her plasma histidine-rich glycoprotein (HRG) level was 50% of the normal level. A low level of plasma HRG was also found in her third daughter. A single nucleotide substitution of T to C was found at nucleotide position 11,438 in exon 6 of the HRG gene from the proband, converting Cys223 to Arg in the second cystatin-like domain. The same mutation was also identified in her third daughter, but not in the other four family members having normal HRG levels or in 50 unrelated healthy Japanese individuals. Expression studies in BHK cells showed that substantial intracellular degradation of the mutant occurred and only about 40% of the recombinant HRG mutant was secreted. These results indicate that congenital HRG deficiency caused by a substitution of Cys223 to Arg is hereditary in this family.
Assuntos
Mutação , Proteínas/genética , Idoso , Sequência de Aminoácidos , Feminino , Humanos , Masculino , Dados de Sequência Molecular , Linhagem , Proteínas/metabolismo , Trombose/genética , Trombose/metabolismoRESUMO
Three thyroidectomized sheep were infused intravenously with porcine calcitonin at a rate of 30 mi.u./h per kg and three sham-operated sheep were infused with vehicle for 5 h. Saliva was collected from the left parotid duct by cannulation for 10 min in every hour. Salivary secretion rates were not changed in either thyroidectomized or sham-operated sheep throughout the experiment. Before the infusion, salivary phosphorus excretion was less in thyroidectomized sheep than in sham-operated animals. Calcitonin infusion increased salivary phosphorus excretion and decreased serum phosphorus concentrations in thyroidectomized sheep. Vehicle infusion did not affect salivary phosphorus excretion in sham-operated sheep. Serum concentrations and salivary excretion of calcium were decreased by calcitonin infusion into thyroidectomized sheep but were not changed in sham-operated sheep infused with vehicle. Calcitonin infusion increased serum parathyroid hormone concentrations in thyroidectomized sheep after the decrease of serum calcium concentrations. However, vehicle infusion did not affect serum parathyroid hormone concentrations in sham-operated sheep. There was little change of cyclic AMP excretion during the experiment in either thyroidectomized or sham-operated sheep. It is concluded that calcitonin increases salivary phosphorus excretion in sheep.
Assuntos
Calcitonina/farmacologia , Cálcio/metabolismo , Fósforo/metabolismo , Saliva/metabolismo , Animais , Cálcio/sangue , Feminino , Hormônio Paratireóideo/sangue , Fósforo/sangue , Ovinos , TireoidectomiaRESUMO
Leucine-enkephalin (L-ENK)-like immunoreactive (L-ENKI) structures in the superior cervical ganglion (SCG) were first examined by using immunoelectron microscopy. L-ENKI neurons formed cell clusters and were small. They were filled with large granular or agranular vesicles and small electron lucent vesicles, and had nuclei that lacked a nucleolus. Since these morphological characteristics are identical to those of the small intensely fluorescent (SIF) cells, L-ENKI cells seemed to be a kind of SIF cell. Two types of L-ENKI fibers were identified, a large type filled with large granular or agranular vesicles and a small type filled with small electron lucent vesicles. The large fibers were located near the L-ENKI perikarya and often could be traced directly to the soma. These fibers remained intact after decentralization of the SCG. These findings indicate that the large L-ENKI fibers are processes of L-ENKI SIF cells. The fibers showed a close apposition to the blood vessels and rarely formed synaptic contact with dendrites of the principal cells. On the other hand, the small L-ENKI fibers were found to originate outside the SCG, because they disappeared after decentralization of the SCG. These L-ENKI fibers frequently formed synaptic contact with the dendrites of the principal cells. Thus, the present study demonstrated the presence of two discrete L-ENKI neuron systems in the SCG of the guinea pig, one an intrinsic SIF system and the other an extrinsic L-ENKI system.
Assuntos
Encefalina Leucina/análise , Gânglios Simpáticos/análise , Neurônios/análise , Animais , Gânglios Simpáticos/ultraestrutura , Cobaias , Histocitoquímica , Técnicas Imunológicas , Masculino , Microscopia Eletrônica , Neurônios/classificação , Neurônios/ultraestruturaRESUMO
The purpose of the present research was to clarify the effect of docosahexaenoic acid (DHA) intake on behavior and plasma catecholamines (CA). In Study 1, 42 students took either DHA-rich oil capsules containing 1.5-1.8 g DHA/d or control oil capsules containing 97% soybean oil plus 3% of another fish oil for 3 mon in a double-blind fashion. They took a psychological test (PF Study) at the start and end of the study. This study started at the end of summer vacation and ended just before the final exams. In the control group, external aggression (aggression against others) in PF Study was significantly increased at the end of the study as compared with that measured at the start (+8.9%), whereas it was not significantly changed in the DHA group (-1.0%). In a similar double-blind study (Study 2), we measured external aggression under nonstressful conditions. External aggression slightly decreased in the control group, whereas there were no significant changes in the DHA group. In Study 3 with 14 students, plasma CA were measured at the start and end of capsule administration period of 2 mon. Subjects were under continuous stress of the final exams that lasted throughout the whole study period. The ratio of plasma epinephrine to norepinephrine concentrations was significantly increased in the DHA group (78%), whereas it stayed at the same level in the control group. In Study 4, mice were fed either DHA-deficient diet or -sufficient diet for 4 wk, and their rearing frequency (an anxiety index) was measured. In the DHA-sufficient group, the rearing frequency was significantly less than in the other group. These effects of DHA intake may be applied to people in an attempt to ameliorate stress-related diseases.
Assuntos
Catecolaminas/sangue , Ácidos Docosa-Hexaenoicos/farmacologia , Estresse Psicológico/sangue , Adulto , Agressão/efeitos dos fármacos , Animais , Método Duplo-Cego , Humanos , Camundongos , Atividade Motora/efeitos dos fármacos , Placebos , Testes Psicológicos , Inquéritos e QuestionáriosRESUMO
We investigated the role of thyroxine on bone growth in energy deficient rats. Rats were fed a normal diet and injected with saline, or a low-energy diet and injected with saline or 0.1, 1, or 5 micrograms/100 g BW of thyroxine for 22 days. Thyroxine injections decreased body weight gain but did not affect bone length and width. The epiphyseal growth plate was thinner and the activities of bone alkaline and acid phosphatase in energy deficient and 5 micrograms/100 g BW thyroxine injected animals were lower than that of the other treatment groups. Serum total thyroxine and triiodothyronine concentrations were increased but somatomedin C concentration was decreased by injecting with 5 micrograms/100 g BW of thyroxine in energy deficient rats.
Assuntos
Desenvolvimento Ósseo/efeitos dos fármacos , Metabolismo Energético , Tiroxina/farmacologia , Fosfatase Ácida/metabolismo , Fosfatase Alcalina/metabolismo , Animais , Dieta , Lâmina de Crescimento/citologia , Lâmina de Crescimento/efeitos dos fármacos , Lâmina de Crescimento/crescimento & desenvolvimento , Fator de Crescimento Insulin-Like I/metabolismo , Masculino , Ratos , Ratos Endogâmicos , Tiroxina/sangue , Tri-Iodotironina/sangueRESUMO
This study investigated the role of thyroxine on bone growth by injecting it into malnourished weaned and suckling rats. Weaned rats were fed a normal diet or a low-protein, low-energy diet, and injected with saline or thyroxine (5 micrograms/100 g BW) for 22 days. Suckling pups were injected with saline or 2 micrograms of thyroxine, and reared by dams fed a normal diet or a low-protein diet for 15 days. Thyroxine injection decreased body weight gain in both growth stages. Bone length, width and weight were not affected by thyroxine injection in weaned rats. Thyroxine injection increased femur length and tail growth, but did not change bone width in suckling rats. The epiphyseal growth plate and zone of hypertrophic cartilage cells were thinner in malnourished rats than in normal rats at both growth stages, but the number of proliferating chondrocytes was greater in suckling rats that received thyroxine injections than in those that received saline injections. The disappearance of trabeculae was observed in both normal and malnourished weaned rats upon thyroxine injection. These results suggested that retarded bone growth due to malnutrition might be independent of thyroid function in both growth stages.
Assuntos
Desenvolvimento Ósseo/efeitos dos fármacos , Deficiência de Proteína/fisiopatologia , Glândula Tireoide/fisiopatologia , Tiroxina/farmacologia , Animais , Fêmur/patologia , Lâmina de Crescimento/patologia , Masculino , Deficiência de Proteína/patologia , Ratos , Ratos Endogâmicos , CaudaRESUMO
The transplacental passage of a single intravenous dose of cefotaxime (CTX), 1,000 mg, was examined in 11 pregnant women undergoing delivery at term by cesarean section. The results were as follows. 1. Measurements by HPLC: After a single 1,000 mg intravenous dose, the maternal blood level of CTX took the following course: 40.1 +/- 3.4 mcg/ml (mean +/- S.E.) at 15 minutes, 22.5 +/- 1.9 mcg/ml at 30 minutes, 10.8 +/- 0.9 mcg/ml at 60 minutes and 3.2 +/- 0.4 mcg/ml at 120 minutes. Slightly high maternal blood levels of 0.7 and 1.1 mcg/ml were obtained at child deliveries made at 200 minutes, but only a trace amount was found at a child delivery at 356 minutes. The CTX level in the umbilical cord blood was comparatively high until the measurement at 222 minutes, and then declined. IN the amniotic fluid, peak levels were observed at 222 to 252 minutes, thereafter decreasing slowly. Desacetyl-CTX, a metabolite of CTX, showed maternal blood levels of 5.8 +/- 0.5 mcg/ml at 15 minutes, 6.3 +/- 0.6 mcg/ml at 30 minutes, 6.8 +/- 0.7 mcg/ml at 60 minutes and 4.9 +/- 0.7 mcg/ml at 120 minutes. Thus, the maternal blood level of desacetyl-CTX showed an increasing tendency until 60 minutes after intravenous administration, and decreased rapidly from 120 minutes onwards. No consistent tendency was noted in desacetyl-CTX levels in the umbilical cord blood and the amniotic fluid. 2. Measurements by bioassay: Maternal blood levels of CTX determined by bioassay were 45.2 +/- 3.3 mcg/ml at 15 minutes, 25.8 +/- 2.2 mcg/ml at 30 minutes, 12.4 +/- 1.0 mcg/ml at 50 minutes adn 4.0 +/- 0.5 mcg/ml at 120 minutes. At the time of child delivery, maternal blood levels of CTX were 1.1 and 2.6 mcg/ml at 200 minutes, and then decreased slowly. Trace levels were noted at 336 minutes and CTX was undetectable at 356 minutes. 3. The HPLC study demonstrated that the level of CTx remained comparatively high in the umbilical cord blood and the amniotic fluid even 6 hours after administration (0.3--0.4 mcg/ml and 4.0 mcg/ml, respectively). This result suggests, in consideration of CTX's MIC values for causative organisms, that CTX will produce a sufficient clinical effect in perinatal infections. No side effects were observed in the mothers or their babies.
Assuntos
Cefotaxima/metabolismo , Troca Materno-Fetal , Placenta/metabolismo , Adulto , Líquido Amniótico/metabolismo , Bioensaio , Cefotaxima/administração & dosagem , Cefotaxima/sangue , Cromatografia Líquida de Alta Pressão , Feminino , Humanos , Injeções Intravenosas , Gravidez , Terceiro Trimestre da GravidezRESUMO
We investigated the prevalence and characteristics of ischemic heart disease especially silent myocardial ischemia (SMI) and arrhythmia in need of careful observation in the exercise stress tests in the Total Health Promotion Plan (THP), which was conducted between 1994-96 for the purpose of measuring cardiopulmonary function. All workers (n = 4,918, 4,426 males) aged 18-60 yr old in an occupational field were studied. Exercise tests with an ergometer were performed by the LOPS protocol, in which the maximal workload was set up as a presumed 70-80% maximal oxygen intake, or STEP (original multistage protocol). ECG changes were evaluated with a CC5 lead. Two hundred and fifteen people refused the study because of a common cold, lumbago and so on. Of 4,703 subjects, 17 with abnormal rest ECG and 19 with probable anginal pain were excluded from the exercise tests. Of 4,667 who underwent the exercise test, 37 (0.79%) had ischemic ECG change, and 155 (3.32%) had striking arrhythmia. These 228 subjects then did a treadmill exercise test with Bruce protocol. Twenty-two (0.47% of 4,703) showed positive ECG change, 9 (0.19%) of 22 had abnormal findings on a 201Tl scan. 8 (0.17%) were diagnosed as SMI (Cohn I), in which the prevalence of hypertension, hyperlipidemia, diabetes mellitus, smoker and positive familial history of ischemic heart disease was greater than that of all subjects. In a 15-30 month follow up, none has developed cardiac accidents. Exercise-induced arrhythmia was detected in 11 (0.23%) subjects. Four were non-sustained ventricular tachycardia without any organic disease, 4 were ventricular arrhythmia based on cardiomyopathy detected by echocardiography, 2 were atrial fibrillation and another was WPW syndrome. It is therefore likely that the ergometer exercise test in THP was effective in preventing sudden death caused by ischemic heart disease or striking arrhythmia.
Assuntos
Arritmias Cardíacas/diagnóstico , Teste de Esforço , Isquemia Miocárdica/diagnóstico , Serviços de Saúde do Trabalhador , Adolescente , Adulto , Arritmias Cardíacas/complicações , Arritmias Cardíacas/etiologia , Morte Súbita Cardíaca/prevenção & controle , Eletrocardiografia , Teste de Esforço/métodos , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Isquemia Miocárdica/complicações , Consumo de Oxigênio , PrevalênciaRESUMO
Aromatase activity in the diencephalon was investigated during the perinatal period in which sex differentiation occurs critically in the rat. 1) Fetal plasma testosterone levels at 17 and 20 days of gestation were slightly higher in the male than in the female. During the critical period after delivery, the hormone levels in the male neonates increased markedly, while it decreased rapidly in the female. 2) Aromatase activity was followed in the diencephalon by measuring the production of estradiol from a labelled substrate, 4-14 C-testosterone. Considerable enzyme activity was detected at 14 days of gestation and it decreased slightly thereafter. The enzyme activity, however, increased drastically during the postnatal critical period. After that it decreased again in both male and female neonates. 3) Aromatase in the telencephalon showed very weak activity with a fluctuation similar to that in the diencephalon. These results suggest that estrogens in the male pups which are specifically converted from testosterone during the critical period may be decisive in sex differentiation of the central nervous system in the rat.
Assuntos
Aromatase/metabolismo , Diencéfalo/enzimologia , Oxirredutases/metabolismo , Fatores Etários , Animais , Animais Recém-Nascidos , Diencéfalo/embriologia , Diencéfalo/fisiologia , Estradiol/biossíntese , Feminino , Feto/enzimologia , Idade Gestacional , Masculino , Gravidez , Ratos , Ratos Endogâmicos , Diferenciação Sexual , Testosterona/sangue , Testosterona/metabolismoAssuntos
Descolamento Prematuro da Placenta/complicações , Coagulação Intravascular Disseminada/tratamento farmacológico , Aprotinina/uso terapêutico , Transfusão de Sangue , Coagulação Intravascular Disseminada/etiologia , Feminino , Fibrinogênio/uso terapêutico , Humanos , Hemorragia Pós-Parto/complicações , GravidezRESUMO
The nuclear DNA contents of atypical mesothelial cells from five patients who had an eosinophilic pleural effusion (EPE) were studied by the use of DAPI (4',6-diamidino-2-phenylindole dihydrochloride) DNA staining. Analysis of the nuclear DNA content revealed a polyploid pattern, with a major peak in the tetraploid region. Using an immunocytochemical technique, the atypical mesothelial cells showed a positive reaction for cytokeratin. In contrast carcinoembryonic antigen (CEA) was always negative in these cells. It is suggested that the atypical mesothelial cells with EPE had a higher rate of proliferation than did the normal mesothelial cells.
Assuntos
Núcleo Celular/química , DNA/análise , Eosinofilia/patologia , Derrame Pleural/patologia , Adulto , Idoso , Biomarcadores , Biomarcadores Tumorais/análise , Antígeno Carcinoembrionário/análise , Carcinoma de Células Escamosas/complicações , Carcinoma de Células Escamosas/patologia , Divisão Celular , Doença de Crohn/complicações , Eosinofilia/etiologia , Epitélio/patologia , Feminino , Humanos , Indóis , Queratinas/análise , Neoplasias Pulmonares/complicações , Neoplasias Pulmonares/patologia , Masculino , Cisto Mediastínico , Glicoproteínas de Membrana/análise , Mucina-1 , Derrame Pleural/etiologia , Pneumonia/complicações , Pneumotórax/etiologia , Pneumotórax/patologia , PoliploidiaRESUMO
The quality of images obtained by a new method of intraoral computed radiography using three types of imaging plates and a drum scanner designed exclusively for use with an imaging plate was investigated. The aperture size for reading of radiographic data from the imaging plate and for film recording was 50, 100, or 150 microns 2. The results indicated that high resolution type imaging plate for mammography produced the best image quality. For each imaging plate, it was also found that the image quality improved as the aperture size was decreased. However, the high resolution type IP even at the aperture size of 100 microns 2 showed diagnostic accuracy as high as the image processing of conventional periapical radiographs. This finding suggests that this new imaging system using an imaging plate at the reading and recording aperture size 100 microns 2 has promise for use in intraoral radiography.
Assuntos
Interpretação de Imagem Radiográfica Assistida por Computador/instrumentação , Radiografia Dentária/instrumentação , Estudos de Avaliação como Assunto , Humanos , Modelos Estruturais , Intensificação de Imagem Radiográfica/instrumentação , Filme para Raios XRESUMO
High serum lipoprotein(a) (Lp(a)) is a risk factor for vascular disorders. Our preliminary observations suggest that, in some patients with coronary heart disease with high serum Lp(a) levels, administration of aspirin reduced Lp(a) levels. Therefore, we aimed to analyze the effects of aspirin on the production of apo(a), the expression of apolipoprotein(a) (apo(a)) mRNA and the transcriptional activity of apo(a) gene promoter. Aspirin (5 mM) reduced the apo(a) levels in culture medium of human hepatocytes and suppressed apo(a) mRNA expression to 73% and 85% of the controls, respectively. Aspirin also reduced the transcriptional activity of apo(a) gene transfected into HepG2 hepatoma cells in a dose-dependent manner, with a maximal effect at 5 mM (44.3 +/- 1.5% of the control). Sodium salicylate (5 mM) also reduced apo(a) gene transcription, whereas indomethacin (10 microM) had no effect. Deletion analysis of apo(a) gene promoter showed that promoter region extending from -30 to +138 is critical for the effect of aspirin. Furthermore, enhanced production, mRNA expression, and gene transcription of apo(a) by interleukin-6 were also inhibited by aspirin. These results demonstrate that aspirin reduces apo(a) production from hepatocytes via reduction of the transcriptional activity of apo(a) gene with suppression of apo(a) mRNA expression. The suppression of apo(a) production by aspirin may at least in part play a role in the anti-atherogenic effect of aspirin in vascular disorders.
Assuntos
Apolipoproteínas A/efeitos dos fármacos , Aspirina/farmacologia , Fígado/efeitos dos fármacos , Apolipoproteínas A/genética , Apolipoproteínas A/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Indometacina/farmacologia , Interleucina-6/farmacologia , Fígado/citologia , Fígado/metabolismo , Luciferases/efeitos dos fármacos , Luciferases/genética , Luciferases/metabolismo , Regiões Promotoras Genéticas/genética , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Proteínas Recombinantes de Fusão/efeitos dos fármacos , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Salicilato de Sódio/farmacologia , Transcrição Gênica/efeitos dos fármacos , Células Tumorais CultivadasRESUMO
A high serum concentration of lipoprotein(a) [Lp(a)] is a significant and independent risk factor for cardiovascular disease. We examined the effects of agents on the transcriptional activity of the apolipoprotein(a) [apo(a)] gene promoter and determined whether drugs identified by this assay would affect the serum concentration of Lp(a) in vivo. All-trans-retinoic acid (ATRA) and interleukin-6 increased the transcriptional activity of the apo(a) gene promoter 2.1- and 2.5-fold, respectively, whereas danazol reduced activity to 76% of the control value. Triiodothyronine had no effect on transcriptional activity. Treatment of two acute promyelocytic leukemia patients with ATRA induced maximal 2.7- and 3.2-fold increases in serum Lp(a) concentrations, respectively. Thus, the in vitro luciferase assay system is capable of identifying agents that affect the serum concentration of Lp(a) and thus may prove beneficial in the screening of new drugs for treatment of individuals with high serum Lp(a) concentrations.
Assuntos
Apolipoproteínas/biossíntese , Apolipoproteínas/genética , Regulação Neoplásica da Expressão Gênica , Leucemia Promielocítica Aguda/tratamento farmacológico , Lipoproteína(a)/sangue , Regiões Promotoras Genéticas , Transcrição Gênica , Adulto , Apoproteína(a) , Sequência de Bases , Linhagem Celular , DNA/sangue , Primers do DNA , Danazol/farmacologia , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Genes Reporter , Hepatoblastoma , Humanos , Interleucina-6/farmacologia , Cinética , Leucemia Promielocítica Aguda/sangue , Neoplasias Hepáticas , Luciferases/biossíntese , Masculino , Pessoa de Meia-Idade , Proteínas Recombinantes de Fusão/biossíntese , Transcrição Gênica/efeitos dos fármacos , Transfecção , Tretinoína/farmacologia , Tretinoína/uso terapêutico , Tri-Iodotironina/farmacologia , Células Tumorais Cultivadas , Vitamina E/farmacologiaRESUMO
OBJECTIVE: To establish a suitable method for measurement of nuclear DNA content in breast tissues from frozen storage after frozen section diagnosis. STUDY DESIGN: For fundamental research, rat liver samples preserved in a deep freezer were used. Four protocols were used (1. fixation with 70% ethanol followed by naked nuclei preparation; 2. fixation with 10% neutral buffered formalin followed by naked nuclei preparation; 3. preparation for naked nuclei prior to fixation with 70% ethanol; and 4. preparation for naked nuclei prior to fixation with 70% neutral buffered formalin). For clinical research, 13 separate fresh frozen breast tissue samples were analyzed after frozen section diagnosis. One contained a malignant phyllodes tumor (MPT) consisting of 2 components, benign epithelial cells and malignant stromal cells; 3 were benign tumors containing fibroadenoma; and 9 cases were carcinomas, consisting of 5 scirrhous, 3 papillotubular and 1 mucinous. RESULTS: Protocols 1, 2 and 3 were not suitable methods for our purpose because remaining cytoplasm or cohesive nuclei were observed. In protocol 4 the cytoplasm was completely undetectable, and nuclei were suitably separated for nuclear DNA content measurement. Benign epithelial cell component nuclei presented a diploid pattern, and the malignant stromal cell component nuclei indicated a euploid pattern in MPT. All 3 cases of benign constituents in fibroadenoma showed a diploid pattern, as did the 3 carcinoma cases (1 mucinous, 1 scirrhous and 1 papillary). Four scirrhous and 2 papillary carcinomas showed an aneuploid pattern. CONCLUSION: Our findings show that it is possible to measure nuclear DNA content of human frozen storage tissues after frozen section diagnosis.
Assuntos
Neoplasias da Mama/genética , Núcleo Celular/genética , DNA de Neoplasias/análise , Citometria de Fluxo/métodos , Secções Congeladas , Animais , Mama/química , Mama/patologia , Mama/ultraestrutura , Neoplasias da Mama/patologia , Neoplasias da Mama/ultraestrutura , Carcinoma/genética , Carcinoma/patologia , Carcinoma/ultraestrutura , Citoplasma , Células Epiteliais/patologia , Células Epiteliais/ultraestrutura , Feminino , Fibroadenoma/genética , Fibroadenoma/patologia , Fibroadenoma/ultraestrutura , Secções Congeladas/normas , Humanos , Fígado/citologia , Fígado/ultraestrutura , Masculino , Microscopia , Tumor Filoide/genética , Tumor Filoide/patologia , Tumor Filoide/ultraestrutura , Ploidias , Ratos , Ratos Endogâmicos F344 , Coloração e Rotulagem , Células Estromais/patologia , Células Estromais/ultraestrutura , Fixação de TecidosRESUMO
OBJECTIVE: To evaluate the difference between benign proliferative lesions of regenerative bone calluses and malignant tumors in rats by simultaneous analysis of nuclear DNA content and size. STUDY DESIGN: Nuclei were stained with DAPI and measured by cytofluorometry. Five regenerative bones and five osteosarcoma samples were used. Two-dimensional dispersion diagrams between relative nuclear DNA content and nuclear size were drawn up and their regression lines were compared. RESULTS: It was possible to distinguish between osteosarcomas and regenerative bones by simultaneous analysis of nuclear size and DNA content during the cell cycle. The results yielded coefficients of correlation and regression. All five regression lines of osteosarcomas were situated dramatically higher on dispersiongrams than those of regenerative bones. CONCLUSION: Differential diagnosis between regenerative lesions and malignant tumors may now be possible using simultaneous measurement of nuclear DNA content and size.