RESUMEN
Adult Teladorsagia circumcincta survival and motility in vitro was examined in a range of different cell culture media, supplements and gas mixes. Under optimum conditions, worms survived for 14 days, exhibiting high motility for 9 days and egg production for 72 h. Optimum conditions involved co-culture of worms with a HeLa cell line in a supplemented cell medium (CEM) and an atmosphere containing 10% CO(2), 5% O(2) 85% N(2), 65% humidity at 37 degrees C. The incubation medium consisted of Minimum Essential Medium with 10% fetal calf serum, 1% non-essential amino acids, 1% glutamax and 1% penicillin-neomycin-streptomycin cocktail mix. Compared with optimum conditions, incubation in CEM alone, cell conditioned CEM, RPMI alone, Medium 199 alone, reduced CO(2) or O(2), or when cells were replaced with Escherichia coli, both survival and motility were reduced. Optimum conditions for adult T. circumcincta maintenance for culture, anthelmintic testing or generation of excretory/secretory products are described.
Asunto(s)
Trichostrongyloidea/crecimiento & desarrollo , Animales , Dióxido de Carbono , Línea Celular , Técnicas de Cocultivo , Medios de Cultivo , Medios de Cultivo Condicionados , Femenino , Células HeLa , Humanos , Masculino , Movimiento , Nitrógeno , Oviposición , Oxígeno , Ovinos , Factores de Tiempo , Trichostrongyloidea/fisiologíaRESUMEN
The occurrence of a thyroid gland superficially placed on the pharyngeal portion of the tongue is rare, but poses problems to the patient and anaesthetist. This report describes a patient with a lingual thyroid and a history of problems associated with it that resulted in admission to the ICU and warnings about future intubation of the larynx. The patient underwent awake tracheal intubation using a standard fibreoptic assisted technique, and was advised that she purchase an appropriate Medic-Alert bracelet.
Asunto(s)
Obstrucción de las Vías Aéreas/etiología , Coristoma/complicaciones , Intubación Intratraqueal/métodos , Glándula Tiroides , Enfermedades de la Lengua/complicaciones , Femenino , Tecnología de Fibra Óptica/métodos , Humanos , Persona de Mediana Edad , Cavidad Nasal , Sistemas de Identificación de PacientesAsunto(s)
Melanocitos/efectos de la radiación , Transcripción Genética/efectos de la radiación , Rayos Ultravioleta , Células Cultivadas , Humanos , Péptidos y Proteínas de Señalización Intracelular , Melanocitos/metabolismo , Antígeno Nuclear de Célula en Proliferación/genética , Proteínas/genética , ARN Mensajero/análisis , Proteinas GADD45RESUMEN
Analysis of the expression of a number of known genes in cultured human cells has revealed UVB-induced changes that may be specific for melanocytic cells. The response of c-fos, p53 and HIV-LTR reporter constructs to UVB and UVC was reduced in MM96L melanoma cells compared to HeLa. Cell cycle arrest produced by UVA, gamma radiation, cisplatin or the antimetabolite deoxyinosine differed from that of UVB. Cell cycle analysis after multiple doses of UVB raised the possibility that UVB-induced pRb depletion could result in increased mutation and thus enhanced tumourigenesis of irradiated melanocytes in skin subjected to a defined pattern of UVB exposure. To extend the analysis of gene expression in cultured melanocytic cells to uncharacterised genes, promoter trap cell clones containing unknown genes 'tagged' by a beta-galactosidase reporter construct were generated from MM96L cells. Altered gene expression in clones treated with a panel of DNA-damaging agents was quantitated by measurement of beta-galactosidase activity. Of the clones containing 'tagged' endogenous promoters induced by UVB, 52% were induced only by UVB and not by other DNA-damaging agents (cisplatin, N-methyl-N-nitro-nitrsoguanidine, fotemustine). One third of the clones were also activated by TPA suggesting that general DNA damage responses involving PKC are activated less frequently than unique pathways of gene activation. Overall, 60% of the 50 clones that responded to the panel of agents were induced by only one of the agents, indicating that a high proportion of genes are induced by agent-specific mechanisms. In the long term, promoter trapping may allow the full repertoire of UVB-inducible genes to be characterised.
Asunto(s)
Melanocitos/efectos de la radiación , Melanoma/etiología , Neoplasias Inducidas por Radiación/patología , Neoplasias Cutáneas/etiología , Rayos Ultravioleta , Antineoplásicos/toxicidad , Ciclo Celular/efectos de los fármacos , Ciclo Celular/efectos de la radiación , Cisplatino/toxicidad , Demecolcina/farmacología , Genes fos , Genes p53 , Duplicado del Terminal Largo de VIH , Células HeLa , Humanos , Hidroxiurea/farmacología , Inosina/análogos & derivados , Inosina/farmacología , Melanocitos/citología , Melanocitos/efectos de los fármacos , Melanoma/patología , Metilnitronitrosoguanidina/toxicidad , Compuestos de Nitrosourea/toxicidad , Compuestos Organofosforados/toxicidad , Proteínas Proto-Oncogénicas c-fos/genética , Proteínas Recombinantes/biosíntesis , Neoplasias Cutáneas/patología , Acetato de Tetradecanoilforbol/toxicidad , Transfección , Células Tumorales Cultivadas , beta-Galactosidasa/genéticaRESUMEN
In human melanocytes and a human melanoma cell line (MM96L), the level of Ku sequence-specific binding to a 37-mer oligonucleotide containing a single E2F-1 binding site of the c-myc promoter (E2cM) significantly decreased 12 24h after cytostatic exposure to 300 J/m2 ultraviolet B radiation (UVB). No UVB-induced loss was found in fibroblasts, while HeLa cells showed an earlier (4 h) but less significant decrease than melanocytic cells. Equitoxic doses of gamma radiation, cisplatin or UVC had little effect on E2cM-specific binding. The loss of Ku binding in MM96L cells was not the result of translocation of Ku or a decrease in Ku protein or DNA-dependent protein kinase activity. The level of E2cM-specific binding in MM96L cells was increased by tunicamycin (2 microg/ml), an inhibitor of N-linked glycosylation, and decreased by the glucosidase inhibitor castanospermine (50 microg/ml). These results, which parallel the reported loss in melanocytes of the cell cycle regulator pRB after UVB, suggest that the DNA binding activity of Ku is affected by post-translational modification and may play a role in regulating the cell cycle response to UVB.
Asunto(s)
Antígenos Nucleares , Proteínas Portadoras , Proteínas de Ciclo Celular , ADN Helicasas , Proteínas de Unión al ADN/metabolismo , Proteínas de Unión al ADN/efectos de la radiación , Melanocitos/efectos de la radiación , Proteínas Nucleares/metabolismo , Proteínas Nucleares/efectos de la radiación , Factores de Transcripción/metabolismo , Anticuerpos Monoclonales/metabolismo , Western Blotting , Ciclo Celular/efectos de la radiación , Factores de Transcripción E2F , Factor de Transcripción E2F1 , Electroforesis en Gel de Poliacrilamida , Inhibidores Enzimáticos/farmacología , Células HeLa , Humanos , Indolizinas/metabolismo , Autoantígeno Ku , Melanoma/metabolismo , Unión Proteica/efectos de la radiación , Proteínas Quinasas/efectos de la radiación , Proteína de Retinoblastoma/efectos de la radiación , Proteína 1 de Unión a Retinoblastoma , Factor de Transcripción DP1 , Células Tumorales Cultivadas , Tunicamicina/metabolismo , Rayos UltravioletaRESUMEN
Azelaic bishydroxamic acid (ABHA), a potent differentiating agent for lymphoid cells, was selectively toxic for 5 human tumor cell lines and transformed human melanocytes and keratinocytes (dose for 37% survival, D37, 30-100 microg/mL) compared with normal cells (melanocytes, fibroblasts; D37 > 300 microg/mL). Dendritic morphology was the only indicator found for increased differentiation, markers for the pigmentation pathway being unchanged or inhibited by ABHA. In contrast to hexamethylene bisacetamide and azelaic acid, ABHA significantly increased the HIV LTR, SV40 and c-fos promoter activities during a 24 hr treatment. Metallothionein promoter activity was enhanced by 5 hr treatment with ABHA in a sensitive melanoma cell line (MM96L) but was inhibited in a more resistant line (HeLa); c-fos promoter activity was inhibited in HeLa during this time. Transcription from a p53 binding response element was inhibited in MM96L by a 24 hr ABHA treatment but enhanced in HeLa. ABHA may represent a structural prototype for designing more potent and selective anti-melanoma agents.
Asunto(s)
Antineoplásicos/farmacología , Ácidos Dicarboxílicos/farmacología , Ácidos Hidroxámicos/farmacología , Melanocitos/efectos de los fármacos , Glicoproteínas de Membrana , Oxidorreductasas , Transcripción Genética/efectos de los fármacos , Acetamidas/farmacología , Línea Celular/efectos de los fármacos , Línea Celular Transformada/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Genes Reporteros , Humanos , Ácidos Hidroxámicos/síntesis química , Interferón Tipo I/análisis , Monofenol Monooxigenasa/análisis , Proteínas/análisis , Transducción de Señal/genética , Células Tumorales Cultivadas/efectos de los fármacosRESUMEN
In human melanocytes and a human melanoma cell line (MM96L), the level of the retinoblastoma gene product (pRB) detected by Western blotting transiently decreased to 55% and 70% of controls respectively 9-12 h after a noncytostatic exposure (75 Jm-2) to UVB (280-315 nm) and to 2% and 14% 48 h after a cytostatic exposure (300 Jm-2). The pRB levels in fibroblasts and HeLa showed minimal loss, and under some conditions increased compared with unirradiated cells. Equitoxic doses of gamma radiation, cisplatin or the antimetabolite deoxyinosine had little effect on pRB levels. UVC (254 nm) was less inhibitory compared with equitoxic UVB. No loss of pRB mRNA was found in MM96L after UVB, nor was pRB protein stability significantly affected. Synthesis of new pRB in MM96L 24 h after UVB was 16% of controls, suggesting that loss of pRB results from a UVB-specific inhibition of translation. Compared with HeLa cells and fibroblasts, MM96L cells exhibited reduced cycle arrest if irradiated when pRB was depleted by a previous UVB exposure. These results suggest a mechanism whereby down-regulation of pRB translation by UVB may play a role in genesis of melanoma.
Asunto(s)
Melanocitos/metabolismo , Melanocitos/efectos de la radiación , Biosíntesis de Proteínas/efectos de la radiación , Proteína de Retinoblastoma/genética , Proteína de Retinoblastoma/metabolismo , Rayos Ultravioleta , Ciclo Celular/efectos de la radiación , Células Cultivadas , Estabilidad de Medicamentos , Humanos , Inmunohistoquímica , Melanocitos/citología , Melanoma/metabolismo , Melanoma/patología , Melanoma/radioterapia , ARN Mensajero/metabolismo , Proteína de Retinoblastoma/biosíntesis , Células Tumorales Cultivadas/efectos de la radiaciónRESUMEN
Human melanoma cell lines have been used to examine the regulation of the tyrosinase (TYR) and tyrosinase-related protein genes TRP-1 and TRP-2 in response to differentiating chemicals and UV irradiation. TRP-1 mRNA levels can be repressed by treatment with the differentiating chemicals DMSO and HMBA. There is little effect of UV irradiation on pigment synthesis by human melanoma cell lines or tyrosinase activity, with variable effects on the levels of the TYR, TRP-1, and TRP-2 gene transcripts. The human TRP-1 gene promoter has been isolated and its activity tested by transient cell transfection to begin an examination of signal transduction mechanisms operating in response to pigmenting and differentiating agents. To identify transcription factors that may be involved in melanocytic gene expression, we studied the N-Oct-3 and N-Oct-5 octamer-binding activities normally expressed in the neuroectodermal cell lineage and which are expressed at high levels in melanoma cells. POU-domain-containing cDNA have been isolated from the A2058 human melanoma cell line that are homologous to the brn-2 gene that encodes N-Oct-3 and N-Oct-5.
Asunto(s)
Proteínas de Unión al ADN , Melanoma/genética , Glicoproteínas de Membrana , Oxidorreductasas , Pigmentación/genética , Factores de Transcripción/metabolismo , Acetamidas/farmacología , Secuencia de Bases , Sitios de Unión , ADN/química , ADN/metabolismo , Dimetilsulfóxido/farmacología , Expresión Génica/efectos de los fármacos , Expresión Génica/efectos de la radiación , Células HeLa , Proteínas de Homeodominio , Humanos , Factores del Dominio POU , Regiones Promotoras Genéticas , Proteínas/genética , ARN Mensajero/biosíntesis , Transducción de Señal , Factores de Transcripción/genética , Transfección , Células Tumorales Cultivadas , Rayos UltravioletaRESUMEN
From a series of 343 nerve biopsies taken by one clinician over a period of 12 years in Nepal, this paper describes the histopathological findings in 153 biopsies from 119 patients suffering from tuberculoid, borderline (dimorphous) or lepromatous leprosy, who were untreated at the time of first presentation and diagnosis. They were taken during the course of other studies, mainly concerned with the mode of transmission of leprosy, and which included biopsies of skin, dartos muscle, nasal mucous membrane and nipple, results of which have already been published. Examination of serial sections by light microscopy revealed a density of cellular infiltration in non-lepromatous cases, or of bacilli in macrophages and Schwann cells in lepromatous cases, which was marked in degree and usually widespread from one end of the biopsy to the other. Intraneural caseation was recorded in four patients with tuberculoid or borderline-tuberculoid leprosy, and many others in this part of the spectrum showed extensive disruption of perineurial and endoneurial structure. In lepromatous patients, the numbers of bacilli in the endoneurial area not infrequently exceeded one thousand per oil immersion field. Although well known to histopathologists familiar with this disease, it is considered that the significance of these findings, in patients presenting for the first time, is not well appreciated by those working in general medicine, neurology, epidemiology, or even in leprosy control.
Asunto(s)
Lepra/patología , Nervios Periféricos/patología , Femenino , Humanos , Lepromina/administración & dosificación , Lepra/diagnóstico , Lepra/microbiología , Masculino , Nepal , Nervios Periféricos/microbiologíaRESUMEN
In a random crossover study in general practice, sixty patients with heart failure were given one week's maintenance treatment with the recommended dosage of Burinex K or Lasix+K. Burinex K appeared to be easier to swallow than the supplement tablets of Lasix+K although the differences were not significant. Patients showed a highly significant preference to take two tablets on one occasion (Burinex K) rather than two different types of tablet on three separate occasions (Lasix+K). Despite pharmacological arguments in favour of stimultaneous versus separate administration of potassium supplement and diuretic, patient acceptibility is of over-riding importance in the long-term.
Asunto(s)
Bumetanida/administración & dosificación , Diuréticos/administración & dosificación , Furosemida/administración & dosificación , Aceptación de la Atención de Salud , Potasio/administración & dosificación , Anciano , Bumetanida/uso terapéutico , Ensayos Clínicos como Asunto , Deglución , Preparaciones de Acción Retardada , Combinación de Medicamentos , Quimioterapia Combinada , Femenino , Furosemida/uso terapéutico , Insuficiencia Cardíaca/tratamiento farmacológico , Humanos , Masculino , Persona de Mediana Edad , Potasio/uso terapéutico , Encuestas y Cuestionarios , ComprimidosRESUMEN
An account is given of the examination of the nasal mucus discharge smears for the presence of Myco. leprae in 322 untreated cases of leprosy seen during a period of one year of which 111 were cases of lepromatous leprosy and remaining 211 were borderline disease. The majority of the patients with lepromatous leprosy had leprosy bacilli in their nasal secretion, often in very great numbers, ans showed a high Morphological Index. Of 41 of the most active cases among the 211 patients with borderline leprosy, only one had Myco. leprae in the nasal mucus. Morphologically normal bacilli are no longer found in nose-blows after 6 months treatment with DDS. It is suggested that the nasal mucus provides a true index of infectivity, and that patients with borderline leprosy are in general not to be regarded as infectious