Synthesis and biological activities of 2 beta-chloro-, 2 beta-fluoro-, and 2 beta-methoxy-1 alpha,25-dihydroxyvitamin D3

Using 1 alpha,2 alpha-oxido-cholesta-5,7-diene-3 beta,25-diol (2) as a starting material, the provitamins of calcitriol with an additional 2 beta-chloro-, 2 beta-fluoro-, and 2 beta-methoxy-substituent (3,4,5) are obtained by transdiaxial opening of the oxirane ring with nucleophiles. An efficient irradiation process is described and used for the synthesis of the 2 beta-substituted calcitriols NS2 (2 beta-Cl), NS6 (2 beta-F), and NS7 (2 beta-OCH3). The affinity of these three vitamin D3 derivatives to the vitamin D receptor (VDR) and was determined. These three A-ring derivatives of 1,25(OH)2D3 were further tested for their proliferation-inhibitory and anti-adipogenic activity and gene regulatoric activity in the vitamin D3-sensitive, murine, mesenchymal cell line C3H10T1/2. The VDR-affinity of the 2 beta-chloro derivative, NS2 (2 beta-Cl), was identical to 1,25(OH)2D3 and its vitamin D binding protein (DBP)-affinity was in the range of 1,25(OH)2D3. NS2 inhibited the proliferation of C3H10T1/2(BMP-4)-cells in the presence of fetal calf serum (FCS) 9 times, and, in the absence of FCS, 111 times lower, as compared with 1,25(OH)2D3. The ID50 dose of adipogenesis-inhibition of NS2 was 13 times higher than the ID50 dose of 1,25(OH)2D3. NS6 (2 beta-F) displayed a slightly higher affinity than 1,25(OH)2D3 to the VDR and DBP-affinity. The proliferation-inhibitory activity in the presence of FCS was 90 times higher, as compared with 1,25(OH)2D3. In the FCS-free proliferation assay NS6 displayed an inhibitory activity in the range of 1,25(OH)2D3. NS6 showed an 5 times higher potency to inhibit (pre)adipocyte-differentiation in C3H10T1/2(BMP-4)-cells than 1,25(OH)2D3. NS7 (2 beta-OCH3) showed the lowest VDR-affinity and the highest DBP-affinity of the tested substances, as compared with 1,25(OH)2D3 (11 times lower and 35 times higher respectively). Its proliferation-inhibitory activity in the FCS-free medium was 9 times and in the FCS-containing assay 67 times lower in comparison with 1,25(OH)2D3. A 1250 times higher NS7-dose was needed to reach the anti-adipogenic potency of 1,25(OH)2D3. All tested substances displayed a similar ability to activate a vitamin D responsive element-regulated reporter gene compared to 1,25(OH)2D3 (NS2 and NS6: 1.3 times higher activity; NS7: 1,4 times lower activity).     

Infrared Spectrum of Ozone in the 4600 and 5300 cm-1 Regions: High Order Accidental Resonances through the Analysis of nu1 + 2nu2 + 3nu3 - nu2, nu1 + 2nu2 + 3nu3, and 4nu1 + nu3 Bands

The very weak bands nu1 + 2nu2 + 3nu3 and 4nu1 + nu3 of 16O3 have been observed for the first time, using the Fourier transform spectrometer (FTS) of Reims and the usual experimental setup providing a large product p x l of approximately 38 Torr x 36 m. The upper levels of these A-type bands which are rather close in energy (they appear respectively at 5291.722 and 5307.790 cm-1) belong to two different sets of interacting polyads. To correctly reproduce the rotation-vibration energy levels and account for the observed perturbations, both bands are treated in a dyad approximation: the (123) state in the Coriolis resonance with the (330) state, and the (401) state in the Coriolis resonance with the (024) state. The assignments of the rotation-vibration levels of the (123) state are confirmed by measurements of line positions of the hot band nu1 + 2nu2 + 3nu3 - nu2 which has also been observed for the first time. The fits are very satisfactory: the r.m.s. deviation for 249 energy levels of the (123) state is 2.4 x 10(-3) cm-1 and is 2.0 x 10(-3) cm-1 for 266 levels of the (401) state. These r.m.s. are near experimental accuracy. Transition moments for the three observed bands are determined from measured line intensities. Copyright 1997 Academic Press. Copyright 1997Academic Press     

The Absorption Spectrum of H2S Between 2150 and 4260 cm-1: Analysis of the Positions and Intensities in the First (2nu2, nu1, and nu3) and Second (3nu2, nu1 + nu2, and nu2 + nu3) Triad Regions

The two triad systems of hydrogen sulfide (2nu2, nu1, and nu3 near 4 μm and 3nu2, nu1 + nu2, and nu2 + nu3 near 2.7 μm) were analyzed using 14 spectra recorded at 0.0056 and 0.011 cm-1 resolution with the McMath Fourier transform spectrometer located at Kitt Peak National Observatory. Experimental upper state levels of H232S, H234S, and H233S were obtained from assigned positions (as high as J = 20 and Ka = 15 for the main isotope). These were fitted to the A-reduced Watson Hamiltonian to determine precise sets of rotational constants through J10 and up to nine Fermi and Coriolis coupling parameters. Intensities of the two H232S triads were modeled with rms values of 2.5%, using the transformed transition moment expansion with 19 terms for 568 intensities of the first triad and 11 terms for the 526 intensities of the second triad. The second derivatives of the dipole moment with respect to normal coordinates were estimated in Debye to be: 22μx = -0.004873 (90); 12μx = 0.01372 (30); and 23μz = 0.01578 (30). This confirmed that for hydrogen sulfide some of the second derivatives are larger than the first derivatives. The calculated line intensities were summed yielding integrated band strengths (in cm-2/atm at 296 K) as follows: 0.3315 for 2nu2, 0.4522 for nu1, 0.1201 for nu3, 0.0303 for 3nu2, 1.820 for nu1 + nu2, and 2.869 for nu2 + nu3. In addition, the hot band transitions were identified in both regions. Finally, a composite database of hydrogen sulfide line parameters was predicted for the 5- to 2.5-μm region. Copyright 1998 Academic Press.     

Simultaneous Analysis of the 2nu2, nu1, and nu3 Bands of Hydrogen Telluride

Spectra of a natural sample of hydrogen telluride as well as a spectrum of monoisotopic H2 130Te have been recorded by means of Fourier transform spectrometry with a resolution of 0.003 cm-1 in the spectral domain 7.5-4.3 μm where it is easy to observe the main absorbing bands nu1 and nu3. We have located and assigned for the first time the 2nu2 band which appears in the lower wavenumber range of the recorded spectral domain near 1700 cm-1. It proved necessary to treat simultaneously the three states (020), (100), and (001). nu1 and nu3 are indeed Coriolis-coupled vibration-rotation bands and it was observed that a few rotational levels of (001) could not be fitted to within their experimental accuracy without considering the second-order Coriolis interaction between the rotational levels of (020) and (001). In this way all the experimental levels were calculated to within the experimental uncertainty, and precise sets of vibrational energies and rotational and coupling constants were obtained for the seven most abundant H2Te isotopic species, namely H2 130Te, H2 128Te, H2 126Te, H2 125Te, H2 124Te, H2 123Te, and H2 122Te. For the most abundant isotopic species H2 130Te the bands centers arenu0 (2nu2) = 1715.9568, nu0 (nu1) = 2065.2709, nu0 (nu3) = 2072.1101 cm-1. Copyright 1997Academic Press     

Assay of matrix metalloproteinases types 1, 2, 3 and 9 in breast cancer

Matrix metalloproteinases (MMPs) are zinc dependent endopeptidases implicated in cancer invasion and metastasis. Gelatin zymography was performed on 84 human breast carcinomas and seven normal breast tissues. The precursor form of MMP-2 (72 kDa) was found in 11 (12%) samples, while its two activated forms, i.e. 62 kDa and 59 kDa, were found in three (6%) and 34 (40%) samples respectively. In contrast to MMP-2, most of the samples (52%) contained MMP-9 in its precursor form. Using ELISA, MMP-1 levels were found in 12% of the samples while MMP-3 levels were found in only 2% of the samples. Levels of MMP-2, -3 and -9 correlated inversely with numbers of nodal metastases. Neither MMP-2 nor -9 levels were significantly related to patient outcome. However, patients with high levels of a 50-kDa gelatinase band after zymography had a significantly better survival than patients with low levels. This species was never observed in normal breast tissue.     

镁砂中Fe2O3、Al2O3、P2O5、TiO2的联合测定

镁砂产品检验中,采用碱熔融法共同预处理样品,在同一母液中,联合测定铁、铝、磷、钛的含量,缩短了检测时间。通过分别控制各元素的显色酸度,各元素线性良好,测得的Fe2O3、Al2O3、P2O5、TiO2的相对标准偏差分别小于1.11%、2.78%、4.84%、5.88%。实验结果表明,方法的准确度和精密度都较高。     

Id1, Id2, and Id3 gene expression in neural cells during development

Id1, Id2, and Id3 mRNA are expressed mainly in the proliferating ependymal cell zone of the mouse brain during embryogenesis. In this study, the expression pattern and cell phenotypes of the Id family mRNA were examined in postnatal and adult rat brain. The expression of Idl and Id3 mRNA in rat brain was observed in the cortex layer 1, corpus callosum, ventricular/subventricular zone (VZ/ SVZ), and the CA1-4 layers of the hippocampus at postnatal day 1 (P1) through P14, whereby it declined at 2 months. In general, the developmental pattern of Idl mRNA coincided with the pattern observed for Id3 mRNA. Similar to Id1 and Id3, Id2 mRNA was highly expressed in the corpus callosum, VZ/SVZ, and the hippocampus. Examination of Id2 mRNA revealed high levels in the cortex and caudate putamen at P1 through P14, whereas a decline was observed in its expression in the adult cortex. In P5 rat cerebellum, all Id mRNA examined were found in the internal granular cell layers; however, at this time point, only Id2 mRNA expression was detected in the differentiating zone of the external granular cell layers, preferentially localizing to adult Purkinje cells. Furthermore, only Id2 mRNA expression in brain was observed in NF+ neurons at P5. Examination of S100alpha+ and GFAP+ astrocytes, revealed the presence of all three mRNAs, whereas the expression of Id2 and Id3 mRNA was absent in 04+ immature oligodendrocytes. These data suggest that the spatial and temporal kinetic patterns during development, as well as cellular specificity, of the Id gene family may play a critical role in neural precursor cell proliferation and cell divergence.     

Synthesis of Hexp-(1-->4)-beta-D-GlcpNAc-(1-->2)-alpha-D-Manp-(1-->O) (CH2)7 CH3 probes for exploration of the substrate specificity of glycosyltransferases: Part II, Hex = 3-O-methyl-beta-D-Gal, 3-deoxy-beta-D-Gal, 3-deoxy-3-fluoro-beta-D-Gal, 3-amino-3-deoxy-beta-D-Gal, beta-D-Gul, alpha-L-Alt, or beta-L-Gal

Seven analogues of the trisaccharide beta-D-Galp-(1-->4)-beta-D-GlcpNAc-(1-->2)-alpha-D-Manp-(1-->O)(CH 2)7CH3 have been synthesized as potential substrates for glycosyltransferases involved in the chain-termination of N-acetyllactosamine-type N-glycans. These compounds include: 3-O-methyl-beta-D-Galp-(1-->4)-beta-D-GlcpNAc-(1-->2)-alpha-D-Manp -(1-->O) (CH2)7CH3, 3-deoxy-beta-D-Galp-(1-->4)-beta-D-GlcpNAc-(1-->2)-alpha-D-Manp-(1 -->O) (CH2)7CH3, 3-deoxy-3-fluoro-beta-D-Galp-(1-->4)-beta-D-GlcpNAc-(1-->2)-alpha-D-M anp- (1-->O)(CH2)7Ch3, 3-amino-3-deoxy-beta-D-Galp-(1-->4)-beta-D-GlcpNAc-(1-->2)-alpha-D-Ma np- (1-->O)(CH2)7CH3, beta-D-Gulp-(1-->4)-beta-D-GlcpNAc-(1-->2)-alpha-D-Manp-(1-- >O)(CH2)7CH3, beta-L-Galp-(1-->4)-beta-D-GlcpNAc-(1-->2)-alpha-D-Manp-(1-->O)(CH 2)7CH3, and alpha-L-Altp-(1-->4)-beta-D-GlcpNAc-(1-->2)-alpha-D-Manp-(1- ->O) (CH2)7CH3. All trisaccharides were obtained by condensation of suitably modified glycosyl donors based on imidates or thioglycosides with the same disaccharide acceptor, octyl 3,4,6-tri-O-benzyl-2-O-(3,6-di-O-benzyl-2-deoxy-2-phthalimido-beta-D- glucopyranosyl)-alpha-D-mannopyranoside, followed by deprotection.