RESUMEN
While the cell wall strictly controls cell size and morphology in bacteria, spheroplasts lack cell walls and can become enlarged in growth medium under optimal conditions. Optimal conditions depend on the bacterial species. We frequently observed extreme enlargement of spheroplasts of the radiation-resistant bacterium Deinococcus grandis in Difco Marine Broth 2216, but not in TGY broth (a commonly used growth medium for Deinococcus). Thorough investigation of media components showed that the presence of Mg2+ or Ca2+ promoted extreme spheroplast enlargement, synthesizing the outer membrane. Our findings strongly suggest that Mg2+ or Ca2+ enlarges spheroplasts, which could change the lipid composition of the spheroplast membrane.
Asunto(s)
Calcio/metabolismo , Deinococcus/crecimiento & desarrollo , Magnesio/metabolismo , Lípidos de la Membrana/metabolismo , Esferoplastos/crecimiento & desarrollo , Medios de Cultivo/metabolismoRESUMEN
On-site monitoring of plasma drug concentrations is required for effective therapies. Recently developed handy biosensors are not yet popular owing to insufficient evaluation of accuracy on clinical samples and the necessity of complicated costly fabrication processes. Here, we approached these bottlenecks via a strategy involving engineeringly unmodified boron-doped diamond (BDD), a sustainable electrochemical material. A sensing system based on a â¼1 cm2 BDD chip, when analysing rat plasma spiked with a molecular-targeting anticancer drug, pazopanib, detected clinically relevant concentrations. The response was stable in 60 sequential measurements on the same chip. In a clinical study, data obtained with a BDD chip were consistent with liquid chromatography-mass spectrometry results. Finally, the portable system with a palm-sized sensor containing the chip analysed â¼40 µL of whole blood from dosed rats within â¼10 min. This approach with the 'reusable' sensor may improve point-of-monitoring systems and personalised medicine while reducing medical costs.
RESUMEN
Although advanced lipidomics technology facilitates quantitation of intracellular lipid components, little is known about the regulation of lipid metabolism in cancer cells. Here, we show that disruption of the Gdpd3 gene encoding a lysophospholipase D enzyme significantly decreased self-renewal capacity in murine chronic myelogenous leukaemia (CML) stem cells in vivo. Sophisticated lipidomics analyses revealed that Gdpd3 deficiency reduced levels of certain lysophosphatidic acids (LPAs) and lipid mediators in CML cells. Loss of Gdpd3 also activated AKT/mTORC1 signalling and cell cycle progression while suppressing Foxo3a/ß-catenin interaction within CML stem cell nuclei. Strikingly, CML stem cells carrying a hypomorphic mutation of Lgr4/Gpr48, which encodes a leucine-rich repeat (LRR)-containing G-protein coupled receptor (GPCR) acting downstream of Gdpd3, displayed inadequate disease-initiating capacity in vivo. Our data showing that lysophospholipid metabolism is required for CML stem cell maintenance in vivo establish a new, biologically significant mechanism of cancer recurrence that is independent of oncogene addiction.
Asunto(s)
Leucemia Mielógena Crónica BCR-ABL Positiva/metabolismo , Hidrolasas Diéster Fosfóricas/metabolismo , Células Madre/metabolismo , Animales , Modelos Animales de Enfermedad , Femenino , Proteína Forkhead Box O3/metabolismo , Lisofosfolípidos/metabolismo , Masculino , Diana Mecanicista del Complejo 1 de la Rapamicina/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Mutación , Recurrencia Local de Neoplasia/metabolismo , Hidrolasas Diéster Fosfóricas/genética , Receptores Acoplados a Proteínas G/genética , Transducción de Señal , beta Catenina/metabolismoRESUMEN
BACKGROUND: The mothers of twins often suffer from sleeplessness. However, little is known about the relation of sleep behaviors between these mothers and their infants. The change of this relation with age has not been reported. The aims of this study are firstly to clarify the sleep behaviors of twin infants and their mothers by using actigraphy (four measurement periods at approximately 4- to 6-week intervals) and secondary to evaluate the relations of sleep behaviors between twin infants and their mothers. METHODS: Five twin pairs and their mothers (first-time mother) were participated in this prospective longitudinal study. Their sleep behaviors were recorded for 7 consecutive days by using an actigraph, when the infants reached a corrected age (CA) of 3-6 weeks, 8-11 weeks, 13-15 weeks, and 17-20 weeks. Sleep status was classified into 3 states: both infants sleeping, only one infant sleeping, and both infants awake. RESULTS: All infants were cobedded. The time awake during the nocturnal period decreased by almost 90 minutes from CA 3-6 weeks to CA 8-11 weeks. Sleep duration in the nocturnal period increased by almost 85 minutes, and the proportion of time with both infants sleeping rapidly increased in the same period. Maternal sleep duration during the period of both infants sleeping was positively correlated with CA. CONCLUSION: This research revealed the transition of sleep behaviors in twin infants and their mothers in early infancy. Cobedding may facilitate more synchronized sleep states of twin infants.
Asunto(s)
Conducta del Lactante , Conducta Materna , Sueño , Gemelos/psicología , Actigrafía , Adulto , Humanos , Lactante , Estudios LongitudinalesRESUMEN
The synthetic ciguatoxin CTX3C has been shown to activate tetrodotoxin (TTX)-sensitive sodium channels (Na(v)1.2, Na(v)1.4, and Na(v)1.5) by accelerating activation kinetics and shifting the activation curve toward hyperpolarization (Yamaoka, K., Inoue, M., Miyahara, H., Miyazaki, K., and Hirama, M. (2004) Br. J. Pharmacol. 142, 879-889). In this study, we further explored the effects of CTX3C on the TTX-resistant sodium channel Na(v)1.8. TTX-resistant channels have been shown to be involved in transducing pain and related sensations (Akopian, A. N., Sivilotti, L., and Wood, J. N. (1996) Nature 379, 257-262). Thus, we hypothesized that ciguatoxin-induced activation of the Na(v)1.8 current would account for the neurological symptoms of ciguatera poisoning. We found that 0.1 mum CTX3C preferentially affected the activation process of the Na(v)1.8 channel compared with those of the Na(v)1.2 and Na(v)1.4 channels. Importantly, without stimulation, 0.1 mum CTX3C induced a large leakage current (I (L)). The conductance of the I (L) calculated relative to the maximum conductance (G (max)) was 10 times larger than that of Na(v)1.2 or Na(v)1.4. To determine the molecular domain of Na(v)1.8 responsible for conferring higher sensitivity to CTX3C, we made two chimeric constructs from Na(v)1.4 and Na(v)1.8. Chimeras containing the N-terminal half of Na(v)1.8 exhibited a large response similar to wild-type Na(v)1.8, indicating that the region conferring high sensitivity to ciguatoxin action is located in the D1 or D2 domains.