RESUMO
BACKGROUND: Parathyroid hormone-related peptide (PTHrP) plays a key role in the development and progression of many tumors. We found that in colorectal cancer (CRC) HCT116 cells, the binding of PTHrP to its receptor PTHR type 1 (PTHR1) activates events associated with an aggressive phenotype. In HCT116 cell xenografts, PTHrP modulates the expression of molecular markers linked to tumor progression. Empirical evidence suggests that the Met receptor is involved in the development and evolution of CRC. Based on these data, we hypothesized that the signaling pathway trigged by PTHrP could be involved in the transactivation of Met and consequently in the aggressive behavior of CRC cells. AIM: To elucidate the relationship among PTHR1, PTHrP, and Met in CRC models. METHODS: For in vitro assays, HCT116 and Caco-2 cells derived from human CRC were incubated in the absence or presence of PTHrP (1-34) (10-8 M). Where indicated, cells were pre-incubated with specific kinase inhibitors or dimethylsulfoxide, the vehicle of the inhibitors. The protein levels were evaluated by Western blot technique. Real-time polymerase chain reaction (RT-qPCR) was carried out to determine the changes in gene expression. Wound healing assay and morphological monitoring were performed to evaluate cell migration and changes related to the epithelial-mesenchymal transition (EMT), respectively. The number of viable HCT116 cells was counted by trypan blue dye exclusion test to evaluate the effects of irinotecan (CPT-11), oxaliplatin (OXA), or doxorubicin (DOXO) with or without PTHrP. For in vivo tests, HCT116 cell xenografts on 6-wk-old male N:NIH (S)_nu mice received daily intratumoral injections of PTHrP (40 µg/kg) in 100 µL phosphate-buffered saline (PBS) or the vehicle (PBS) as a control during 20 d. Humanitarian slaughter was carried out and the tumors were removed, weighed, and fixed in a 4% formaldehyde solution for subsequent treatment by immunoassays. To evaluate the expression of molecular markers in human tumor samples, we studied 23 specimens obtained from CRC patients which were treated at the Hospital Interzonal de Graves y Agudos Dr. José Penna (Bahía Blanca, Buenos Aires, Argentina) and the Hospital Provincial de Neuquén (Neuquén, Neuquén, Argentina) from January 1990 to December 2007. Seven cases with normal colorectal tissues were assigned to the control group. Tumor tissue samples and clinical histories of patients were analyzed. Paraffin-embedded blocks from primary tumors were reviewed by hematoxylin-eosin staining technique; subsequently, representative histological samples were selected from each patient. From each paraffin block, tumor sections were stained for immunohistochemical detection. The statistical significance of differences was analyzed using proper statistical analysis. The results were considered statistically significant at P < 0.05. RESULTS: By Western blot analysis and using total Met antibody, we found that PTHrP regulated Met expression in HCT116 cells but not in Caco-2 cells. In HCT116 cells, Met protein levels increased at 30 min (P < 0.01) and at 20 h (P < 0.01) whereas the levels diminished at 3 min (P < 0.05), 10 min (P < 0.01), and 1 h to 5 h (P < 0.01) of PTHrP treatment. Using an active Met antibody, we found that where the protein levels of total Met decreased (3 min, 10 min, and 60 min of PTHrP exposure), the status of phosphorylated/activated Met increased (P < 0.01) at the same time, suggesting that Met undergoes proteasomal degradation after its phosphorylation/activation by PTHrP. The increment of its protein level after these decreases (at 30 min and 20 h) suggests a modulation of Met expression by PTHrP in order to improve Met levels and this idea is supported by our observation that the cytokine increased Met mRNA levels at least at 15 min in HCT116 cells as revealed by RT-qPCR analysis (P < 0.05). We then proceeded to evaluate the signaling pathways that mediate the phosphorylation/ activation of Met induced by PTHrP in HCT116 cells. By Western blot technique, we observed that PP1, a specific inhibitor of the activation of the proto-oncogene protein tyrosine kinase Src, blocked the effect of PTHrP on Met phosphorylation (P < 0.05). Furthermore, the selective inhibition of the ERK 1/2 mitogen-activated protein kinase (ERK 1/2 MAPK) using PD98059 and the p38 MAPK using SB203580 diminished the effect of PTHrP on Met phosphorylation/activation (P < 0.05). Using SU11274, the specific inhibitor of Met activation, and trypan blue dye exclusion test, Western blot, wound healing assay, and morphological analysis with a microscope, we observed the reversal of cell events induced by PTHrP such as cell proliferation (P < 0.05), migration (P < 0.05), and the EMT program (P < 0.01) in HCT116 cells. Also, PTHrP favored the chemoresistance to CPT-11 (P < 0.001), OXA (P < 0.01), and DOXO (P < 0.01) through the Met pathway. Taken together, these findings suggest that Met activated by PTHrP participates in events associated with the aggressive phenotype of CRC cells. By immunohistochemical analysis, we found that PTHrP in HCT116 cell xenografts enhanced the protein expression of Met (0.190 ± 0.014) compared to tumors from control mice (0.110 ± 0.012; P < 0.05) and of its own receptor (2.27 ± 0.20) compared to tumors from control mice (1.98 ± 0.14; P < 0.01). Finally, assuming that the changes in the expression of PTHrP and its receptor are directly correlated, we investigated the expression of both Met and PTHR1 in biopsies of CRC patients by immunohistochemical analysis. Comparing histologically differentiated tumors with respect to those less differentiated, we found that the labeling intensity for Met and PTHR1 increased and diminished in a gradual manner, respectively (P < 0.05). CONCLUSION: PTHrP acts through the Met pathway in CRC cells and regulates Met expression in a CRC animal model. More basic and clinical studies are needed to further evaluate the PTHrP/Met relationship.
Assuntos
Neoplasias Colorretais , Proteína Relacionada ao Hormônio Paratireóideo , Animais , Células CACO-2 , Movimento Celular , Proliferação de Células , Neoplasias Colorretais/tratamento farmacológico , Neoplasias Colorretais/genética , Neoplasias Colorretais/patologia , Humanos , Irinotecano , Masculino , Camundongos , Proteína Relacionada ao Hormônio Paratireóideo/genética , Proteína Relacionada ao Hormônio Paratireóideo/metabolismo , Proteína Relacionada ao Hormônio Paratireóideo/farmacologia , Azul Tripano/farmacologiaRESUMO
INTRODUCTION: It is hypothesized that increased macrophage migration inhibitory factor (MIF) expression may contribute to diabetic nephropathy (DN) pathogenesis. The aim of the present study was to investigate the renal effects of MIF inhibition in a diabetic experimental model. METHODS: Eighteen male Wistar rats (230 ± 20 g) were divided into three groups: 1) control, 2) diabetic (STZ, 50 mg/kg, dissolved in saline, ip), 3) diabetic + MIF antagonist (p425, 1 mg/kg per day, ip, on the 21th day, for 21 consecutive days). The treatment started since we founwd a significant increase in urine albumin excretion (UAE) rate in the diabetic rats in comparison with the control rats. The rats were kept individually in metabolic cages (8 AM-2 PM) and urine samples were collected in the 21 and 42th day. At the end, blood and tissue samples were collected for biochemical (BS, UPE, urine GAG, BUN, Cr, Na, and K) and histological analyses. RESULTS: The results of this study showed that MIF antagonist (p425) significantly decreased urine protein and GAG excretion, urine protein/creatinine ratio, and serum BUN and Cr in the streptozotocin-induced DN in the rats. Pathological changes were significantly alleviated in the MIF antagonist (p425)-administered DN rats. CONCLUSION: Collectively, these data suggested that MIF antagonist (p425) was able to protect against functional and histopathological injury in the DN.
Assuntos
Diabetes Mellitus Experimental/patologia , Nefropatias Diabéticas/tratamento farmacológico , Oxirredutases Intramoleculares/antagonistas & inibidores , Fatores Inibidores da Migração de Macrófagos/antagonistas & inibidores , Substâncias Protetoras/farmacologia , Substâncias Protetoras/uso terapêutico , Azul Tripano/farmacologia , Azul Tripano/uso terapêutico , Albuminúria/tratamento farmacológico , Animais , Glicemia , Creatinina/sangue , Creatinina/urina , Diabetes Mellitus Experimental/sangue , Diabetes Mellitus Experimental/induzido quimicamente , Diabetes Mellitus Experimental/urina , Nefropatias Diabéticas/sangue , Nefropatias Diabéticas/patologia , Nefropatias Diabéticas/urina , Modelos Animais de Doenças , Glicosaminoglicanos/urina , Rim/patologia , Ativação de Macrófagos , Masculino , Ratos , Ratos Wistar , Estreptozocina/farmacologiaRESUMO
Purpose: The present experimental study aimed to investigate the effects of intracameral trypan blue (TB) on oxidative stress parameters and apoptosis in corneal tissue. Methods: Thirty rats were randomly assigned to three groups of 10 rats each: the sham group (Group 1); control group (Group 2); and treatment group (Group 3). The control group was administered 0.01 cc of balanced salt solution. The treatment group was administered 0.006 mg/0.01 cc of TB. The total antioxidant status (TAS) and total oxidant status (TOS) in corneal tissue and blood were measured and the oxidative stress index (OSI) was calculated. Finally, corneal tissue histopathology was evaluated using staining for caspase-3 and -8, and apoptotic activity was examined. Results: The TAS, TOS and OSI levels in the blood samples were not significantly different (p>0.05 for all). Compared with the sham and control groups, the TOS and OSI levels in corneal tissue were significantly different in the treatment group (p<0.05 for all). No significant difference was observed between the sham group and the control group (p>0.05). Immunohistochemical staining for caspase-3 and caspase-8 demonstrated higher apoptotic activity in the TB group than in the sham and control groups. Conclusion: The present study showed that intracameral TB injection is safe systematically but may be toxic to corneal tissue, as demonstrated using oxidative stress parameters and histopathological evaluation. .
Objetivo: Este estudo experimental tem como objetivo investigar os efeitos do azul de tripan intracameral (TB) sobre parâmetros de estresse oxidativo e apoptose no tecido da córnea. Métodos: Trinta ratos foram divididos aleatoriamente em três grupos de 10 animais cada: grupo simulação (Grupo 1); grupo controle (Grupo 2); e grupo tratamento (Grupo 3). No grupo controle foi administrado 0,01 cc de solução salina balanceada (BSS). No grupo tratamento foi administrado 0,006 mg/0,01 cm de TB. O estado antioxidante total ( TAS) e estado oxidante total ( TOS) no tecido da córnea e sangue foram medidos e o índice de estresse oxidativo (OSI) foi calculado. Finalmente, histopatologia do tecido da córnea foi avaliada por meio da coloração para caspase-3 e -8; atividade apoptótica também foi examinada. Resultados: Os níveis de TAS, TOS e OSI das amostras de sangue não foram significativamente diferentes (p>0,05 para todos). Em comparação com os grupos simulação e controle, os níveis de TOS e OSI no tecido da córnea foram significativamente diferentes no grupo tratamento (p<0,05 para todos). Não houve diferença significativa entre o grupo simulção e o grupo controle (p>0,05). A coloração imuno-histoquímica com a caspase-3 e caspase-8 demonstrou maior atividade apoptótica no grupo tratamento do que nos grupos controle e simulação. Conclusão: Este estudo mostrou que a injeção intracameral TB é segura sistematicamente, mas pode ser tóxica ao tecido da córnea, como demonstrado através de parâmetros de estresse oxidativo e avaliação histopatológica. .
Assuntos
Animais , Masculino , Apoptose/efeitos dos fármacos , Corantes/farmacologia , Córnea/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Azul Tripano/farmacologia , Antioxidantes/análise , /análise , /análise , Estudos de Viabilidade , Imuno-Histoquímica , Injeções Intraoculares , Oxidantes , Distribuição Aleatória , Ratos Wistar , Valores de Referência , Reprodutibilidade dos Testes , Azul Tripano/administração & dosagemRESUMO
The compounds 6-dimethylaminopurine and cycloheximide promote the successful production of cloned mammals and have been used in the development of embryos produced by somatic cell nuclear transfer. This study investigated the effects of 6-dimethylaminopurine and cycloheximide in vitro, using the thiazolyl blue tetrazolium bromide colorimetric assay to assess cytotoxicity, the trypan blue exclusion assay to assess cell viability, the comet assay to assess genotoxicity, and the micronucleus test with cytokinesis block to test mutagenicity. In addition, the comet assay and the micronucleus test were also performed on peripheral blood cells of 54 male Swiss mice, 35 g each, to assess the effects of the compounds in vivo. The results indicated that both 6-dimethylaminopurine and cycloheximide, at the concentrations and doses tested, were cytotoxic in vitro and genotoxic and mutagenic in vitro and in vivo, altered the nuclear division index in vitro, but did not diminish cell viability in vitro. Considering that alterations in DNA play important roles in mutagenesis, carcinogenesis, and morphofunctional teratogenesis and reduce embryonic viability, this study indicated that 6-dimethylaminopurine and cycloheximide utilized in the process of mammalian cloning may be responsible for the low embryo viability commonly seen in nuclear transfer after implantation in utero.
Assuntos
Adenina/análogos & derivados , Clonagem de Organismos/métodos , Ensaio Cometa , Cicloeximida/toxicidade , Mutagênicos/toxicidade , Adenina/toxicidade , Animais , Técnicas de Cultura de Células , Sobrevivência Celular/efeitos dos fármacos , Corantes , Citocinese/efeitos dos fármacos , Células Hep G2/efeitos dos fármacos , Humanos , Masculino , Mamíferos , Camundongos , Testes para Micronúcleos , Testes de Mutagenicidade , Técnicas de Transferência Nuclear , Sais de Tetrazólio/farmacologia , Tiazóis/farmacologia , Azul Tripano/farmacologiaRESUMO
The compounds 6-dimethylaminopurine and cycloheximide promote the successful production of cloned mammals and have been used in the development of embryos produced by somatic cell nuclear transfer. This study investigated the effects of 6-dimethylaminopurine and cycloheximide in vitro, using the thiazolyl blue tetrazolium bromide colorimetric assay to assess cytotoxicity, the trypan blue exclusion assay to assess cell viability, the comet assay to assess genotoxicity, and the micronucleus test with cytokinesis block to test mutagenicity. In addition, the comet assay and the micronucleus test were also performed on peripheral blood cells of 54 male Swiss mice, 35 g each, to assess the effects of the compounds in vivo. The results indicated that both 6-dimethylaminopurine and cycloheximide, at the concentrations and doses tested, were cytotoxic in vitro and genotoxic and mutagenic in vitro and in vivo, altered the nuclear division index in vitro, but did not diminish cell viability in vitro. Considering that alterations in DNA play important roles in mutagenesis, carcinogenesis, and morphofunctional teratogenesis and reduce embryonic viability, this study indicated that 6-dimethylaminopurine and cycloheximide utilized in the process of mammalian cloning may be responsible for the low embryo viability commonly seen in nuclear transfer after implantation in utero.
Assuntos
Animais , Humanos , Masculino , Camundongos , Adenina/análogos & derivados , Ensaio Cometa , Clonagem de Organismos/métodos , Cicloeximida/toxicidade , Mutagênicos/toxicidade , Adenina/toxicidade , Técnicas de Cultura de Células , Corantes , Sobrevivência Celular/efeitos dos fármacos , Citocinese/efeitos dos fármacos , /efeitos dos fármacos , Mamíferos , Testes para Micronúcleos , Testes de Mutagenicidade , Técnicas de Transferência Nuclear , Sais de Tetrazólio/farmacologia , Tiazóis/farmacologia , Azul Tripano/farmacologiaRESUMO
PURPOSE: The present experimental study aimed to investigate the effects of intracameral trypan blue (TB) on oxidative stress parameters and apoptosis in corneal tissue. METHODS: Thirty rats were randomly assigned to three groups of 10 rats each: the sham group (Group 1); control group (Group 2); and treatment group (Group 3). The control group was administered 0.01 cc of balanced salt solution. The treatment group was administered 0.006 mg/0.01 cc of TB. The total antioxidant status (TAS) and total oxidant status (TOS) in corneal tissue and blood were measured and the oxidative stress index (OSI) was calculated. Finally, corneal tissue histopathology was evaluated using staining for caspase-3 and -8, and apoptotic activity was examined. RESULTS: The TAS, TOS and OSI levels in the blood samples were not significantly different (p>0.05 for all). Compared with the sham and control groups, the TOS and OSI levels in corneal tissue were significantly different in the treatment group (p<0.05 for all). No significant difference was observed between the sham group and the control group (p>0.05). Immunohistochemical staining for caspase-3 and caspase-8 demonstrated higher apoptotic activity in the TB group than in the sham and control groups. CONCLUSION: The present study showed that intracameral TB injection is safe systematically but may be toxic to corneal tissue, as demonstrated using oxidative stress parameters and histopathological evaluation.
Assuntos
Apoptose/efeitos dos fármacos , Corantes/farmacologia , Córnea/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Azul Tripano/farmacologia , Animais , Antioxidantes/análise , Caspase 3/análise , Caspase 8/análise , Estudos de Viabilidade , Imuno-Histoquímica , Injeções Intraoculares , Masculino , Oxidantes , Distribuição Aleatória , Ratos Wistar , Valores de Referência , Reprodutibilidade dos Testes , Azul Tripano/administração & dosagemRESUMO
PURPOSE: To evaluate the ultrastructural effect of trypan blue 0.1% staining for capsulorhexis on lens epithelial cells (LECs) and capsules. SETTING: Division of Ophthalmology, University of São Paulo, São Paulo, Brazil. METHODS: Before capsulorhexis, patients were randomly assigned to 1 of 2 groups. Trypan blue 0.1% staining was performed in the treatment group. No trypan blue was used in the control group. Samples of capsules with LECs were fixed and analyzed with routine optical microscopy techniques, immunohistochemistry for beclin-1 expression (a marker of autophagy), terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling to detect apoptosis, and transmission electron microscopy (TEM). Morphometric analyses were performed, and the 2 sets of data were compared. RESULTS: Each group comprised 15 patients. Cell death by autophagy and apoptosis was observed in the treatment group but not in the control group. The TEM images of subcapsular epithelium cells showed mitochondrial rupture, dilation of the cisterns of the endoplasmic reticulum, increased cytoplasmic and nuclear electron density, and abnormalities in the nuclear profile of trypan blue-stained cells. Morphometric analysis showed statistically significant differences between the 2 groups in the longest nuclear axes and the ratio between the total nuclear perimeter and the cell area (P = .03). The difference in capsule thickness between groups was not significant. CONCLUSION: Trypan blue caused LEC death, which supports the hypothesis that staining with trypan blue 0.1% can help reduce the incidence of posterior capsule opacification after cataract surgery. FINANCIAL DISCLOSURE: No author has a financial or proprietary interest in any material or method mentioned.
Assuntos
Capsulorrexe , Corantes/farmacologia , Cápsula do Cristalino/ultraestrutura , Cristalino/ultraestrutura , Azul Tripano/farmacologia , Apoptose/efeitos dos fármacos , Proteínas Reguladoras de Apoptose/metabolismo , Proteína Beclina-1 , Fragmentação do DNA , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Células Epiteliais/ultraestrutura , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Marcação In Situ das Extremidades Cortadas , Cápsula do Cristalino/efeitos dos fármacos , Cristalino/efeitos dos fármacos , Cristalino/metabolismo , Proteínas de Membrana/metabolismo , Microscopia Eletrônica de Transmissão , Microscopia de Fluorescência , Facoemulsificação , Estudos Prospectivos , Coloração e RotulagemRESUMO
The present work describes a simple method for direct drug administration into the dorsal root ganglion (DRG) in anesthetized rats. This technique does not involve surgery, is easy to learn and allows behavioral testing within minutes after the injection. Based on landmarks that target the L5 DRG, an orifice was created with a guide needle through which a specially designed needle was inserted for solution injection. Its introduction into the ganglia was ensured by the triggering of an ipsilateral hindpaw reflex. The precision of the technique was checked by injections of the biological dye Pontamine Sky Blue (PSB) or C14-labeled arginine. There was no leakage of the dye to the surrounding tissues after a single 4 microl or three successive 2.5 microl injections (at 30-min intervals). Moreover, identical effects were observed with prostaglandin E2 (PGE2), morphine or glibenclamide injected intraplantarly or in the DRG, thus confirming the precision of the method and suggesting that the ganglion cells and peripheral nociceptors may display similar receptor population.
Assuntos
Gânglios Espinais/efeitos dos fármacos , Microinjeções/instrumentação , Microinjeções/métodos , Nociceptores/efeitos dos fármacos , Analgésicos Opioides/farmacologia , Anestesia , Animais , Arginina/farmacologia , Radioisótopos de Carbono , Corantes/farmacologia , Dinoprostona/farmacologia , Glibureto/farmacologia , Membro Posterior , Hipoglicemiantes/farmacologia , Masculino , Morfina/farmacologia , Agulhas , Ratos , Ratos Wistar , Reflexo/efeitos dos fármacos , Reflexo/fisiologia , Azul Tripano/farmacologiaRESUMO
PURPOSE: To determine the potential risk of contamination of a trypan blue bottle (TB) after first use and after being stored under different temperature and humidity conditions, as well as to identify possible contamination factors, most frequently involved microorganisms and simultaneously evaluate bacteriostatic and bactericide properties of the dye. METHODS: An experimental and prospective study was carried out, in which 30 TB bottles were divided into 3 groups (A: control, B: refrigerator storage and C: cabinet storage). The dye was suctioned and cultivated in agar blood plates and Sabouraud agar tube. In group A, TB was cultivated immediately after the opening of the bottles (temperature zero - T0), in groups B and C cultivation occurred in T0, T1 (1 day), T2 (2 days), T7 (7 days) and T10 (10 days) after the opening of the bottles. On the 10th day, groups B and C bottles were also submitted to scraping of their inner walls after opening. Concurrently, the inhibitory action measurement test was conducted on the TB dye for the study of bacteriostatic and bactericidal activity. RESULTS: Cultivation procedures conducted in T0 presented no contamination. Among T1 and T10, added to the scraping there was only one contaminated bottle stored in the refrigerator. The encountered microorganism was Aspergillus niger. It has been proven that the dye does not show bactericide and bacteriostatic properties against the bacteria which were tested. CONCLUSIONS: Under this study's conditions there was no contamination of the bottles stored in cabinets and 1 bottle (10%) stored in the refrigerator showed contamination after opening and initial use. The source of contamination may possibly be the outer part of the product. TB does not show bactericidal and bacteriostatic properties against the tested bacteria and in the applied concentration.
Assuntos
Aspergillus niger/isolamento & purificação , Corantes , Contaminação de Medicamentos , Armazenamento de Medicamentos , Azul Tripano , Corantes/farmacologia , Escherichia coli/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Estudos Prospectivos , Pseudomonas aeruginosa/efeitos dos fármacos , Fatores de Risco , Staphylococcus aureus/efeitos dos fármacos , Azul Tripano/farmacologiaRESUMO
OBJETIVOS: Determinar o potencial risco de contaminacão do frasco de azul de tripano (AT) depois de utilizado pela primeira vez e estocado em diferentes condicões de temperatura e umidade, assim como identificar os possíveis fatores de contaminacão, microrganismos mais freqüentemente envolvidos e simultaneamente avaliar as propriedades bacteriostáticas e bactericidas do corante. MÉTODOS: Realizado estudo experimental, prospectivo, em que 30 frascos de AT foram divididos em três grupos (A: controle, B: armazenamento em geladeira e C: armazenamento em armário). O corante era aspirado e semeado em placas de ágar sangue e tubo de ágar Sabouraud. No grupo A o AT foi semeado apenas logo após a abertura dos frascos (tempo zero - T0), nos grupos B e C ocorreu semeadura nos T0, T1 (1 dia), T2 (2 dias), T7 (7 dias) e T10 (10 dias) após abertura dos frascos. No 10º dia os frascos dos grupos B e C também foram submetidos a um raspado do lado interno do frasco após abertura. Concomitantemente foi realizado teste de acão inibitória do corante AT para estudo da atividade bacteriostática e bactericida. RESULTADOS: As semeaduras realizadas no T0 não apresentaram contaminacão. Entre os T1 e T10 mais o raspado houve apenas 1 frasco contaminado armazenado em geladeira. O microrganismo encontrado foi o Aspergillus niger. Foi comprovado que o corante não apresenta acão bactericida e bacteriostática para as bactérias testadas. CONCLUSÕES: Nas condicões do estudo não houve contaminacão dos frascos armazenados em armário e 1 frasco (10 por cento) armazenado em geladeira apresentou contaminacão após abertura e uso inicial. A fonte de contaminacão talvez seja o lado externo do produto. O AT não apresenta propriedades bactericidas e bacteriostáticas para as bactérias testadas e na concentracão utilizada.
Assuntos
Aspergillus niger/isolamento & purificação , Corantes , Contaminação de Medicamentos , Armazenamento de Medicamentos , Azul Tripano , Corantes/farmacologia , Escherichia coli/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Estudos Prospectivos , Pseudomonas aeruginosa/efeitos dos fármacos , Fatores de Risco , Staphylococcus aureus/efeitos dos fármacos , Azul Tripano/farmacologiaRESUMO
Nonimmune rosette formation, an in vitro measure of cellular immunity, was evaluated in normal newborn infants. Active rosette formation in 14 specimens of cord blood was 18.9 +/- 4.8% compared to 28.1 +/- 5.2% in 15 adult control samples (p less than 0.05). Total rosette formation in 13 cord blood samples was 33.3 +/- 7.6% compared to 55.1 +/- 6.5% in 15 adult control specimens (p less than 0.05). Tritiated thymidine uptake from phytohemagglutinin stimulation was comparable in cord blood and adult control lymphocytes. The importance of these findings is discussed in light of other recent reports suggesting that cord blood thymic derived T lymphocytes may have reduced immune capability when compared to adult lymphocytes.