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•Uniform glibenclamide (GCM) thin films were fabricated using spin coating approach.•Progress of amide to imide tautomerisation were probed in thin films.•Explanation of the ...autocatalytic nature of the tautomerism was provided.•The tautomerism phenomenon was studied in different parts of GCM molecules.
The uniform thin films with variable thicknesses (d = 49, 120, 220 nm) of active pharmaceutical ingredient (API) glibenclamide (GCM) was spin-coated and investigated using broadband dielectric, grazing incident FTIR spectroscopies, atomic force microscopy, and ellipsometry. Data analysis revealed that nanoconfined systems consist of a mixture of amide and imidic acid forms of this pharmaceutical, wherein the ratios of both tautomeric forms in the thin films were different with respect to the molten supercooled bulk system. Moreover, changes in the populations of glibenclamide tautomers, i.e. higher amide to imides ratio in the spatially restricted API with respect to the bulk sample, had a strong impact on the character of the proton transfer reaction. In this context, the kinetic curves constructed on the base of infrared data for the bulk system follow the sigmoidal shape, characteristic for the autocatalytic reaction, while results obtained for the confined samples provide exponential character and indicate first-order transformation. This allows hypothesizing that the autocatalytic nature of the tautomerism in the bulk sample is most likely related to the formation of the amide tautomers which further catalyze the progress of imide-amide transformation. Our results are the first studies showing that the change in the thickness of the film may affect the properties and isomerization kinetics in a pharmaceutical systems. Finally, our data open a new perspective for developing new drug delivery systems.
The NEMO-3 results for the double-
β
decay of
150
Nd to the 0
1
+
and 2
1
+
excited states of
150
Sm are reported. The data recorded during 5.25 year with 36.6 g of the isotope
150
Nd are used in the ...analysis. The signal of the
2
ν
β
β
transition to the 0
1
+
excited state is detected with a statistical significance exceeding 5
σ
. The half-life is measured to be
T
1
/
2
2
ν
β
β
(
0
1
+
)
=
1
.
11
-
0.14
+
0.19
stat
-
0.15
+
0.17
syst
×
10
20
year, which is the most precise value that has been measured to date. 90% confidence-level limits are set for the other decay modes. For the
2
ν
β
β
decay to the 2
1
+
level the limit is
T
1
/
2
2
ν
β
β
(
2
1
+
)
>
2.42
×
10
20
year
. The limits on the
0
ν
β
β
decay to the 0
1
+
and 2
1
+
levels of
150
Sm are significantly improved to
T
1
/
2
0
ν
β
β
(
0
1
+
)
>
1.36
×
10
22
year
and
T
1
/
2
0
ν
β
β
(
2
1
+
)
>
1.26
×
10
22
year
.
Using data from the NEMO-3 experiment, we have measured the two-neutrino double beta decay ( Formula omitted) half-life of Formula omittedSe as Formula omitted y under the single-state dominance ...hypothesis for this nuclear transition. The corresponding nuclear matrix element is Formula omitted. In addition, a search for neutrinoless double beta decay ( Formula omitted) using 0.93 kg of Formula omittedSe observed for a total of 5.25 y has been conducted and no evidence for a signal has been found. The resulting half-life limit of Formula omitted for the light neutrino exchange mechanism leads to a constraint on the effective Majorana neutrino mass of Formula omitted, where the range reflects Formula omitted nuclear matrix element values from different calculations. Furthermore, constraints on lepton number violating parameters for other Formula omitted mechanisms, such as right-handed currents, majoron emission and R-parity violating supersymmetry modes have been set.
Three-dimensional homology models of cytochromes P450 (P450) 2B1 and P450 3A4 have been utilized along with site-directed mutagenesis to elucidate the molecular determinants of substrate specificity. ...Most of the key residues identified in 2B enzymes fall within five substrate recognition sites (SRSs) and have counterparts in bacterial P450 residues that regulate substrate binding or access. Docking of inhibitors into 2B models has provided a plausible explanation for changes in susceptibility to mechanism-based inactivation that accompany particular amino acid side-chain replacements. These studies provide a basis for predicting drug interactions due to P450 inhibition and for rational inhibitor design. In addition, the location of P450 3A4 residues capable of influencing homotropic stimulation by substrates and heterotropic stimulation by flavonoids has been identified. Steroid hydroxylation by the wild-type enzyme exhibits sigmoidal kinetics, indicative of positive cooperativity. Based on the 3A4 model and single-site mutants, a double mutant in SRS-2 has been constructed that exhibits normal Michaelis-Menten kinetics. Results of modeling and mutagenesis studies suggest that the substrate and effector bind at adjacent sites within a single large cavity in P450 3A4. A thorough understanding of the location and structural requirements of the substrate-binding and effector sites in cytochrome P450 3A4 should prove valuable in rationalizing and predicting interactions among the multitude of drugs and other compounds that bind to the enzyme.
The full data set of the NEMO-3 experiment has been used to measure the half-life of the two-neutrino double beta decay of Formula omittedMo to the ground state of Formula omittedRu, Formula omitted ...year. The two-electron energy sum, single electron energy spectra and distribution of the angle between the electrons are presented with an unprecedented statistics of Formula omitted events and a signal-to-background ratio of Formula omitted 80. Clear evidence for the Single State Dominance model is found for this nuclear transition. Limits on Majoron emitting neutrinoless double beta decay modes with spectral indices of Formula omitted, as well as constraints on Lorentz invariance violation and on the bosonic neutrino contribution to the two-neutrino double beta decay mode are obtained.
Steroidogenesis is initiated by the conversion of cholesterol to pregnenolone by mitochondrial cytochrome P450scc cholesterol, reduced-adrenal-ferredoxin:oxygen oxido-reductase (side-chain-cleaving); ...EC 1.14.15.6. Several subsequent steroidal conversions occur in the endoplasmic reticulum (ER), but the last step in the production of glucocorticoids and mineralocorticoids again occurs in the mitochondria. Although cellular compartmentalization of steroidogenic enzymes appears to be a feature of all steroidogenic pathways, some reports indicate that cholesterol can be converted to pregnenolone outside the mitochondria. To investigate whether P450scc can function outside the mitochondria, we constructed vectors producing P450scc and various fusion enzymes of P450scc with electron-transport proteins and directed their expression to either the ER or the mitochondria. Whether targeted to mitochondria or to the ER, plasmid vectors encoding P450scc and fusion proteins of P450scc with either mitochondrial or microsomal electron-transport proteins produced immunodetectable protein. When expressed in mitochondria, all of these constructions converted 22-hydroxycholesterol to pregnenolone, but when expressed in the ER none of them produced pregnenolone. These results show that P450scc can function only in the mitochondria. Furthermore, it appears to be the mitochondrial environment that is required, rather than the specific mitochondrial electron-transport intermediates.
The three-dimensional structure of human cytochrome P450 3A4 was modeled based on crystallographic coordinates of four bacterial P450s; P450 BM-3, P450cam, P450terp, and P450eryF. The P450 3A4 ...sequence was aligned to those of the known proteins using a structure-based alignment of P450 BM-3, P450cam, P450terp, and P450eryF. The coordinates of the model were then calculated using a consensus strategy, and the final structure was optimized in the presence of water. The P450 3A4 model resembles P450 BM-3 the most, but the B' helix is similar to that of P450eryF, which leads to an enlarged active site when compared with P450 BM-3, P450cam, and P450terp. The 3A4 residues equivalent to known substrate contact residues of the bacterial proteins and key residues of rat P450 2B1 are located in the active site or the substrate access channel. Docking of progesterone into the P450 3A4 model demonstrated that the substrate bound in a 6 beta-orientation can interact with a number of active site residues, such as 114, 119, 301, 304, 305, 309, 370, 373, and 479, through hydrophobic interactions. The active site of the enzyme can also accommodate erythromycin, which, in addition to the residues listed for progesterone, also contacts residues 101, 104, 105, 214, 215, 217, 218, 374, and 478. The majority of 3A4 residues which interact with progesterone and/or erythromycin possess their equivalents in key residues of P450 2B enzymes, except for residues 297, 480 and 482, which do not contact either substrate in P450 3A4. The results from docking of progesterone and erythromycin into the enzyme model make it possible to pinpoint residues which may be important for 3A4 function and to target them for site-directed mutagenesis.
Abstract The full data set of the NEMO-3 experiment has been used to measure the half-life of the two-neutrino double beta decay of $$^{100}$$ 100 Mo to the ground state of $$^{100}$$ 100 Ru, ...$$T_{1/2} = \left 6.81 \pm 0.01\,\left( \text{ stat }\right) ^{+0.38}_{-0.40}\,\left( \text{ syst }\right) \right \times 10^{18}$$ T1/2=6.81±0.01stat-0.40+0.38syst×1018 year. The two-electron energy sum, single electron energy spectra and distribution of the angle between the electrons are presented with an unprecedented statistics of $$5\times 10^5$$ 5×105 events and a signal-to-background ratio of $$\sim $$ ∼ 80. Clear evidence for the Single State Dominance model is found for this nuclear transition. Limits on Majoron emitting neutrinoless double beta decay modes with spectral indices of $$\mathrm{n}=2,3,7$$ n=2,3,7 , as well as constraints on Lorentz invariance violation and on the bosonic neutrino contribution to the two-neutrino double beta decay mode are obtained.
A variable gain amplifier for 900-MHz applications has been designed and fabricated in a BiCMOS process with f/sub T/ = 24 GHz. The amplifier has linear-in-dB gain control with a 50-dB control range. ...The maximum gain is 28 dB and the third-order output intercept point (OIP3) is 13.7 dBm. The gain is achieved in one gain stage with a cascoded output. The amplifier bias network and the gain-control circuitry are temperature compensated for temperature-independent gain at any gain setting. The bias network also uses a feedback loop to cancel out undesired low frequencies present at the radio-frequency input. The maximum output power is +10 dBm and the output 1-dB compression point is +8.7 dBm. Active chip area is 0.1 mm/sup 2/. The amplifier is packaged in a SOT-363 and consumes 30 mA from a 2.8-V supply.
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