The pharmacokinetics of flomoxef in serum and in the mucosal tissue of the middle ear and mastoid were studied in 9 patients undergoing tympanoplasties. All patients received 1 g of flomoxef intravenously. Flomoxef levels in serum and in mucosal tissue were determined by a bioassay method. The peak value of mean concentrations of flomoxef in the mucosal tissue was 30.3 +/- 11.7 micrograms/ml at 10 min after the administrations. Pharmacokinetic analyses showed that the concentration of flomoxef in the mucosal tissue was over 1.56 micrograms/ml (which is the MIC90 for the common pathogens of otitis media) for more than 2 h and decreased parallel with serum concentration with a half-life of about 40 min.
Abstract Microtubule (MT) dynamics are modulated through the coordinated action of various MT-associated proteins (MAPs). However, the regulatory mechanisms underlying MT dynamics remain unclear. We show that the MAP7 family protein Map7D2 stabilizes MTs to control cell motility and neurite outgrowth. Map7D2 directly bound to MTs through its N-terminal half and stabilized MTs in vitro . Map7D2 localized prominently to the centrosome and partially on MTs in mouse N1-E115 neuronal cells, which expresses two of the four MAP7 family members, Map7D2 and Map7D1. Map7D2 loss decreased the resistance to the MT-destabilizing agent, nocodazole without affecting acetylated/detyrosinated stable MTs, suggesting that Map7D2 stabilizes MTs via direct binding. In addition, Map7D2 loss increased the rate of random cell migration and neurite outgrowth, presumably by disturbing the balance between MT stabilization and destabilization. Map7D1 exhibited similar subcellular localization and gene knock-down phenotypes to Map7D2. However, in contrast to Map7D2, Map7D1 was required for the maintenance of acetylated stable MTs. Taken together, our data suggest that Map7D2 and Map7D1 facilitate MT stabilization through distinct mechanisms in cell motility and neurite outgrowth.
Mechanical forces influence cellular proliferation, differentiation, tissue morphogenesis, and functional expression within the body. To comprehend the impact of these forces on living organisms, their quantification is essential. This study introduces a novel microdifferential pressure measurement device tailored for cellular-scale pressure assessments. The device comprises a glass substrate and a microchannel constructed of polydimethylsiloxane, polytetrafluoroethylene tubes, a glass capillary, and a microsyringe pump. This device obviates the need for electrical measurements, relying solely on the displacement of ultrapure water within the microchannel to assess the micropressure in embryos. First, the device was subjected to arbitrary pressures, and the relationship between the pressure and the displacement of ultrapure water in the microchannel was determined. Calibration results showed that the displacement dx [μm] could be calculated from the pressure P [Pa] using the equation dx = 0.36 P. The coefficient of determination was shown to be 0.87, indicating a linear response. When utilized to measure brain ventricular pressure in mouse embryos, the fabricated device yielded an average pressure reading of 1313 ± 640 Pa. This device can facilitate the measurement of pressure within microcavities in living tissues and other areas requiring precise and localized pressure evaluations.
In vertebrate embryos, formation of anterior neural structures requires suppression of Wnt signals emanating from the paraxial mesoderm and midbrain territory. In Six3(-/-) mice, the prosencephalon was severely truncated, and the expression of Wnt1 was rostrally expanded, a finding that indicates that the mutant head was posteriorized. Ectopic expression of Six3 in chick and fish embryos, together with the use of in vivo and in vitro DNA-binding assays, allowed us to determine that Six3 is a direct negative regulator of Wnt1 expression. These results, together with those of phenotypic rescue of headless/tcf3 zebrafish mutants by mouse Six3, demonstrate that regionalization of the vertebrate forebrain involves repression of Wnt1 expression by Six3 within the anterior neuroectoderm. Furthermore, these results support the hypothesis that a Wnt signal gradient specifies posterior fates in the anterior neural plate.
Regionalization of the embryonic brain is achieved through multi-step processes that operate sequentially and/or simultaneously. Localized sources of various signaling molecules act as organizing centers that pattern neighboring fields to create molecularly distinct domains. We investigated the mechanisms underlying the regionally distinct competence for two such organizing signals, Fibroblast growth factor 8 (Fgf8) and Sonic hedgehog (Shh), using chick embryos. First, we demonstrated that FGF receptor 1 (Fgfr1) and Fgfr3, expressed differentially in the developing brain, possess an equivalent potential to induce the regionally distinct Fgf8-responsive genes, depending on the anterior-posterior dimension of the brain. Next we found that homeodomain transcription factors Six3 and Irx3 can alter the regional responses to both Fgf8 and Shh in the forebrain. Six3 confers the ability to express Bf1, a gene essential for the telencephalon and eye development, and Nkx2.1, which is required for development of the hypothalamus. In contrast, Irx3 confers the ability to express En2 and Nkx6.1 in response to Fgf8 and Shh, respectively. Furthermore, an alteration in the region-specific response to Fgf8 upon misexpression of Irx3 resulted in transformation of diencephalic and possibly telencephalic tissues into the optic tectum. Finally, we demonstrated that Six3 and Irx3 can mutually repress their expression, which may contribute to the establishment of their complementary expression domains in the neural plate. These repressive interactions are specific, as Six3 did not repress Gbx2, and Irx3 did not disturb Otx2 expression. These findings provide evidence that the early embryonic forebrain is demarcated into two domains with distinct genetic programs, which argues against the authentic telen-diencephalic subdivision.
MS Cohtin (MS-C) is a controlled-release tablet of morphine sulfate developed by Muhdipharma AG, Switzerland, for the purpose of maintaining the blood concentration of morphine in an effective analgesic range by administration every 12 hr.In this study, the pharmacokihetics of morphine (M), and its metabolites, morphine-6-glucuronide (M-6-G) and morphine-3-glucuronide (M-3-G), were investigated in patients with cancer pain at steady state following oral administration of MS-C.MS-C was administered every 12 hr, with the dose being adjusted so as to control each patient's pain.In the patients administered MS-C at doses of 20, 30, or 40 mg every 12 hr, the plasma concentrations of M, M-6-G, and M-3-G showed broad hill-like patterns over a period of 12 hr with the peaks occurring at 4 hr. The plasma cohcentrations were found to be almost proportional to the doses.Cmax and AUC values of M, M-6-G, and M-3-G were also almost proportional to the doses, and t1/2 (kab), t1/2 (kel), τ, and Tmax values were comparable between the doses.Total urihary recovery rates from 24-hr urine and the recovery rates for M, M-6-G, and M-3-G were comparable between the doses.These results indicate that there is no significant change in the pharmacokinetics following MS-C administration at various doses in cancer patients.
Cells are able to adhere selectively to particular cell types. This property of cells is considered to play an important role in development of the nervous system. For example, the selective adhesiveness might be essential for developing neurons to seek and bind to the particular target cells, or it might work for sorting different types of neurons and glias to establish the highly ordered stereotypic cell arrangement in neural tissues during development. In fact, it is known that disaggregated neural cells can reconstitute the original tissue-like structures when reaggregated by allocating themselves in a tissue-specific pattern (Fujisawa 1971). It is likely that such cell behaviors are regulated at least partly by the molecules involved in cell-cell adhesion, although many other factors, such as cell-matrix adhesion molecules, cell migration activators or inhibitors, chemotactic factors, and growth factors, might also be involved (Dodd and Jessel 1988).
The Pax6 gene plays an important role in eye morphogenesis throughout the animal kingdom. The Pax6 gene and its homologue could form ectopic eyes by targeted expression in Drosophila and Xenopus. Thus, this gene is a master gene for the eye morphogenesis at least in these animals. In the early development of the vertebrate eye, Pax6 is required for the instruction of multipotential progenitor cells of the neural retina (NR). Primitive retinal pigment epithelial (RPE) cells are able to switch their phenotype and differentiate into NR under exogenous intervention, including treatment with fibroblast growth factors (FGFs), and surgical removal of endogenous NR. However, the molecular basis of phenotypic switching is still controversial. Here, we show that Pax6 alone is sufficient to induce transdifferentiation of ectopic NR from RPE cells without addition of FGFs or surgical manipulation. Pax6-mediated transdifferentiation can be induced even at later stages of development. Both in vivo and in vitro studies show that the Pax6 lies downstream of FGF signaling, highlighting the central roles of Pax6 in NR transdifferentiation. Our results provide an evidence of retinogenic potential of nearly mature RPE and a cue for new therapeutic approaches to regenerate functional NR in patients with a visual loss.
