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Ionizing radiation induces expression and binding activity of the nuclear factor kappa B.

Recent studies have demonstrated that treatment of mammalian cells with ionizing radiation is associated with activation of gene expression. Although the signal transduction pathways stimulated by ionizing radiation remain unclear, our previous findings indicate that radiation induces specific genes at the transcriptional level. The present work has examined the effects of ionizing radiation on the transcription factor NF-kappa B. The results demonstrate that ionizing radiation activates DNA binding of nuclear factor (NF)kappa B. This effect was detectable at 2 grays (Gy) and reached a maximum at 5-20 Gy. At a dose of 20 Gy, the increase in NF-kappa B binding activity was maximal at 2-4 h and then declined to pretreatment levels. The results also demonstrate that ionizing radiation transiently increases NF-kappa B mRNA levels. However, the finding that induction of NF-kappa B binding to DNA occurs in the presence of cycloheximide indicates that ionizing radiation activates preexisting NF-kappa B protein. NF-kappa B exists as a cytoplasmic protein before activation. Thus, our results suggest that ionizing radiation induces transduction pathways which include cytoplasmic signaling events.

Base Sequence↗

An overview of some recent developments in ionization methods for mass spectrometry.

An overview of some recent advances in ionization sources for mass spectrometry is presented. Limitations were set so that the overview covers ionization techniques relevant to organic and biological analysis that have appeared in the literature since the year 2000. No effort is made to be comprehensive. Rather, a broad sweep overview of author-subjective highlights among a wide variety of sources is presented. These ionization sources include electron ionization, chemical ionization, various atmospheric plasma ionization sources, laser desorption sources, sonic spray and electrospray ionization sources.

Atmospheric Pressure↗

Ionizing radiation induces, via generation of reactive oxygen intermediates, intercellular adhesion molecule-1 (ICAM-1) gene transcription and NF kappa B-like binding activity in the ICAM-1 transcriptional regulatory region.

Ionizing radiation produces reactive oxygen intermediates in mammalian tissues and may serve as a model system for the investigation of the biologic effects of free radicals. We have previously shown that the adhesion molecule ICAM-1 is induced by ionizing radiation, and here we have investigated the molecular mechanisms responsible. ICAM-1 mRNA and cell surface expression was induced in HeLa and HaCaT cells after exposure to ionizing radiation. This induction was blocked by preincubation with the antioxidants PDTC and N-acetyl cysteine. ICAM-1 promoter activity was assessed by transiently transfecting HeLa cells with CAT-reporter gene constructs containing sequential ICAM-1 5' deletions. ICAM-1 5' fragments -1162/+1 (relative to the transcription start site) and -277/+1 displayed increased promoter activity when cells were exposed to ionizing radiation, but no induction was seen in a -182/+1 construct associating positions -277 to around -182 with inducibility by ionizing radiation. Nuclear extracts from HaCaT cells were tested in mobility shift assays using an NF kappa B-like binding site of the ICAM-1 5' region (positions -186/-177). There was marked enhancement of DNA-protein complex forming in extracts from irradiated versus untreated cells. Incubation of cells with antioxidants prior to irradiation prevented the radiation-dependent increase in complex formation. We conclude that reactive oxygen intermediates are involved in ICAM-1 induction by ionizing radiation. The ionizing radiation-induced, antioxidant-inhibitable binding at the ICAM-1 NF kappa B-like binding site is consistent with the view that NF kappa B is a pro-oxidant transcription factor.

Binding Sites↗

Systematic analysis of desorption chemical ionization (DCI) mass spectra of nucleic acids--7.

The positive ion and negative ion pyrolysis mass spectra of the herring sperm DNA have been studied using desorption chemical ionization. The positive ion desorption chemical ionization spectra have been produced with CH4, i-C4H10, NH3, HCl and Cl2; the negative ones with N2O/CH4, N2O/i-C4H10, Cl2, CCl4, HCl and via electron capture. These spectra have been compared with the electron impact ionization spectra. We have observed an important increase of sensitivity when negative ionization has replaced the positive ionization mode. The series of diagnostic ions resulting from direct chemical ionization belong to the family of base + reagent ion X [BH + X] and base + X - HX ion [B]. Their abundance has increased considerably compared to the electron impact spectra. The application of these new diagnostic ions in nucleic acid studies is interesting especially for the much higher abundance of the usually weak dG fragment ion obtained in the negative ionization mode. The dG-base segment of the DNA is the most nucleophilic centre of the whole nucleic acid and is implicated in numerous important biochemical reactions involving, for example, proteins.

Animals↗

Dissolution and ionization of warfarin.

It has been shown in recent studies that warfarin exists in the solid state and in some nonaqueous solvents as a cyclic hemiketal. The present study was undertaken to investigate the ionization and ionization kinetics of warfarin, to confirm the probable existence of the cyclic hemiketal in aqueous solution, and to determine the possible consequences of the cyclic hemiketal to acyclic enol equilibrium and ionization kinetics on the dissolution rate of warfarin. The equilibrium aqueous solubility of un-ionized warfarin acid at 25 degrees C and ionic strength 0.5 (with potassium chloride) was found to be 1.28 X 10(-5) M, and its observed macroscopic pK alpha was 5.03-5.06, depending on the method of determination. By comparing the aqueous pK alpha of warfarin to phenprocoumin, a hydroxycoumarin that cannot exist in the cyclic hemiketal form, the hemiketal-acyclic enol ratio was estimated to be approximately 20:1. By stop-flow spectrophotometry, the ionization rate of warfarin (pH 3.5 jumped to pH 6.5) was found to have t1/2 less than 1-2 X 10(-3) s. The dissolution rate of warfarin from a rotating disk (600 rpm), as a function of pH, was measured under nonbuffered but pH-stat conditions (mu = 0.5 with potassium chloride). The pH-dissolution rate profile for warfarin agreed with that calculated from an equation derived previously to describe the dissolution of instantaneous ionizing acids, i.e., the profile was not perturbed from that expected from an acid of aqueous solubility 1.28 X 10(-5) M (un-ionized form) and pK alpha 5.06.

Chemistry, Pharmaceutical↗

Total and ionized serum magnesium in critically ill patients.

OBJECTIVE: To assess the alterations in total serum magnesium (tsMg) and ionized serum magnesium (Mg(2+)) and their association with prognosis in critically ill patients. DESIGN AND SETTING: Prospective, cohort study in the intensive care unit (ICU) of a university teaching hospital. PATIENTS: Adult patients admitted to the ICU without previous factors influencing magnesium homeostasis were included during a 6-month period. MEASUREMENTS AND RESULTS: One hundred forty four patients were included. Mean age was 60.6+/-15.4 years; mean APACHE II score was 12.6+/-6.9. Blood samples were collected in the first 24 h after ICU admission and again on the second, third, and last days of stay in the ICU. At ICU admission 52.5% had total hypomagnesemia and 13.5% total hypermagnesemia; with respect to the Mg(2+) 9.7% showed ionized hypomagnesemia and 23.6% ionized hypermagnesemia. Patients who developed ionized hypermagnesemia had higher mortality than patients without ionized hypermagnesemia development (P=0.04). A moderate correlation between tsMg and Mg(2+) concentrations was found; however, a number of patients with total hypomagnesemia (69-85% during the study) had ionized normomagnesemia. The measure of agreement between tsMg and Mg(2+) levels was poor. CONCLUSIONS: Magnesium alterations are frequently found in critically ill patients. The usually determined tsMg levels are not a reflection of Mg(2+) levels. Development of ionized hypermagnesemia is associated with prognosis.

APACHE↗

Ionization states in the microenvironment of solid dosage forms: effect of formulation variables and processing.

PURPOSE: Evaluation of the effect of formulation composition and processing variables on the microenvironment in solid dosage forms, based on ionization of indicator probes. MATERIALS AND METHODS: Sulfonephthalein indicators were intimately mixed with individual excipients, binary excipient mixtures or multi-component blends by the solvent deposition method. Diffuse reflectance visible spectroscopy of these solids provided a measure of indicator ionization extent. Indicator solution studies yielded equations relating solution pH to the ratio of the absorbance signals of the ionized to that of the unionized form, for each indicator. These equations and the spectral data of the indicator-treated solids were used to calculate an acidity function, 'pH(eq)' for the solids. The ionization of incorporated probes was also monitored during various stages of simulated pharmaceutical processing viz. wet and dry mixing. RESULTS: The pH(eq) provided a measure of the physicochemical environment experienced by the probe in the solid. The surface nature of formulation components and their surface area available for interaction influenced the overall properties of the final blend. The extent of probe ionization varied at different stages of a simulated wet mixing-drying process. The pH of the excipient suspension was not a good predictor of the probe ionization in the final dried solid. Indicator ionization is expected to be influenced by the microenvironmental acidity, polarity and ionic strength. Individual excipient properties contributed to the overall microenvironment in powder mixtures even when dry mixed at low water contents. CONCLUSIONS: The environment experienced by a drug in the final solid dosage form will be influenced by the nature of the excipients, the extent of their surfaces available for interaction, surface modification during processing and the amount and nature of solvent used.

Algorithms↗

Loss of ions in cavity ionization chambers.

Ion losses due to initial recombination, volume recombination, and back diffusion were each determined by measurements and calculations for different size cylindrical ionization chambers and spherical ionization chambers. By measuring signal currents from these ionization chambers irradiated with (60)Co gamma rays, two groups of ion losses were obtained. (Group 1) Ion loss due to initial recombination and diffusion, which changes proportionally to the inverse of the voltage applied to the ionization chambers; (and group 2) ion loss due to volume recombination, which changes proportionally to the inverse of the square of the applied voltage. The diffusion loss was obtained separately by computing electric field distributions in the ionization chambers. It was found that diffusion loss is larger than initial recombination loss for the cylindrical ionization chambers and vise versa for the spherical ionization chambers.

Journal Article↗

Are acidic and basic groups in buried proteins predicted to be ionized?

Ionizable residues play essential roles in proteins, modulating protein stability, fold and function. Asp, Glu, Arg, and Lys make up about a quarter of the residues in an average protein. Multi-conformation continuum electrostatic (MCCE) calculations were used to predict the ionization states of all acidic and basic residues in 490 proteins. Of all 36,192 ionizable residues, 93.5% were predicted to be ionized. Thirty-five percent have lost 4.08 kcal/mol solvation energy (DeltaDeltaG(rxn)) sufficient to shift a pK(a) by three pH units in the absence of other interactions and 17% have DeltaDeltaG(rxn) sufficient to shift pK(a) by five pH units. Overall 85% of these buried residues (DeltaDeltaG(rxn)>5DeltapK units) are ionized, including 92% of the Arg, 86% of the Asp, 77% of the Glu, and 75% of the Lys. Ion-pair interactions stabilize the ionization of both acids and bases. The backbone dipoles stabilize anions more than cations. The interactions with polar side-chains are also different for acids and bases. Asn and Gln stabilize all charges, Ser and Thr stabilize only acids while Tyr rarely stabilize Lys. Thus, hydroxyls are better hydrogen bond donors than acceptors. Buried ionized residues are more likely to be conserved than those on the surface. There are 3.95 residues buried per 100 residues in an average protein.

Amino Acids, Acidic↗

Application of surface ionization methods for highly sensitive and selective analysis of benzodiazepine derivatives.

In this work, the results of studying the possibilities of surface ionization mass spectrometry (SI/MS) and atmosphere pressure thermodesorption surface ionization (APTDSI) spectroscopy for high-sensitivity and selective detection and identification of psychotropic preparations of benzodiazepine derivatives--medazepam, diazepam and chlordiazepoxide--are presented. It has been established that the SI mass spectra of benzodiazepines have a small number of lines and are significantly different from those obtained by electron ionization (EI) and that the molecules of benzodiazepines can be ionized by surface ionization with high efficiency (> or = 100 times and more than by EI) and the current density increases from diazepam to medazepam. It has been found that the APTDSI spectra of benzodiazepine derivatives have two characteristic maxima connected with energy of sublimation and desorption. The ionization efficiency is several C/g (Coulomb per gram), the linear range of the concentration dependence is 3-4 orders of a magnitude. The results obtained by the SI/MS and APTDSI methods have been compared with those obtained by the conventional TLC and GC/MS with electron ionization.

Benzodiazepines↗

Monitoring of serum ionized magnesium in neurosurgical intensive care unit: preliminary results.

BACKGROUND: Our purpose was to determine the values for serum ionized magnesium (Mg) concentrations in traumatic brain injury and its effect on the prognostic scores of patients. METHODS: We prospectively measured serum ionized magnesium concentrations in 30 patients that were classified into three groups (severe, moderate, mild) by Glasgow Coma Scale Score. Serum ionized magnesium concentrations were measured during posttraumatic 5 days. Thirty patients with head trauma were followed in a neurosurgical intensive care unit with monitoring serum ionized magnesium concentrations. All patients were treated conservatively. RESULTS: We found significant difference of serum ionized magnesium concentrations when we compared all groups with each other (p<0.001). CONCLUSIONS: Based on this clinical preliminary study, traumatic brain injury is associated with graded deficit in serum ionized magnesium concentrations. Thus, measurement of serum ionized magnesium concentrations can be used as a clinical marker in traumatic brain injury.

Adult↗

Sex steroid hormones modulate serum ionized magnesium and calcium levels throughout the menstrual cycle in women.

OBJECTIVE: To determine the serum concentrations of the sex steroid hormones with respect to the concentrations of the biologically active fractions of magnesium and calcium during the different phases of the menstrual cycle. DESIGN: Controlled clinical study. SETTING: An academic research environment. PATIENT(S): Six parous and four nulliparous healthy cycling female volunteers. MAIN OUTCOME MEASURE(S): Concentrations of the sex steroid hormones estrogen, progesterone, and testosterone as well as the ionized Ca and Mg levels were measured in the serum of normal cycling women during five different stages: the menstrual, early follicular, late follicular, ovulatory (ovulatory/early luteal), and luteal phases. RESULT(S): In each woman, there was a comparatively high ionized Mg level coincident with the early follicular phase, a statistically significant decrease in ionized Mg around the time of ovulation, a significant decrease in ionized and total Mg when the serum progesterone concentration peaked, and a significant increase in the serum Ca2+/Mg2+ ratio at both the ovulatory and luteal phases. In addition, a decrease in ionized Mg was found with increased testosterone levels. CONCLUSION(S): Healthy women of reproductive age demonstrate recurring cycling of ionized Mg and cyclic alterations in the ionized Ca to Mg ratio in their serum. The changes in serum concentrations of these important physiologically active cations, in the range at which they occur, can affect such entities as the vasculature, synaptic transmission, and excitation-secretion coupling and thus can produce the well-known premenstrual syndromes during the luteal phase in women who are somewhat deficient in Mg or in those who have an unusually increased Ca2+/Mg2+ ratio.

Adult↗

Serum ionized magnesium levels in normal and preeclamptic gestation.

OBJECTIVE: To compare serum levels of ionized and total magnesium with those of ionized calcium, total calcium, sodium, and potassium over the course of pregnancy in normal women and in women who develop preeclampsia. METHODS: We collected venous serum samples from 31 pregnant women during their first, second, and third trimesters. Gestational ages ranged from 6 to 37 weeks. Samples were analyzed for ionized and total magnesium, ionized and total calcium, sodium, and potassium using a biomedical chemistry analyzer. Data were analyzed with repeated-measures analysis of variance and two-way repeated-measured analysis of variance. RESULTS: In 22 normal pregnant women, both serum ionized and total magnesium levels decreased significantly with increasing gestational age. No changes in sodium, potassium, or ionized or total calcium were observed. Nine of the 31 subjects developed preeclampsia by term; serum total magnesium levels decreased significantly by the second trimester in these women compared with those of normal pregnant women. CONCLUSION: Our results provide evidence of decreases in ionized and total magnesium levels with increasing gestational age during normal pregnancy, as well as evidence of a magnesium disturbance in women who later develop preeclampsia. Future studies of magnesium balance in women at risk for developing complications of pregnancy are indicated.

Adult↗

The effects of electron and chemical ionization modes on the MS profiling of whole bacteria.

Free fatty acid profiling of whole bacteria [Francisella tularensis, Brucella melitensis, Yersinia pestis, Bacillus anthracis (vegetative and sporulated), and Bacillus cereus] was carried out with direct probe mass spectrometry under 70-eV electron ionization (EI) and isobutane chemical ionization in both the positive (CI+) and negative modes (CI-). Electron ionization produced spectra that contained molecular ions and fragment ions from various free fatty acids. Spectra acquired with isobutane chemical ionization in the positive mode yielded molecular ions of free fatty acids as well as ions from other bacterial compounds not observed under EI conditions. Spectra obtained with negative chemical ionization did not contain as much taxonomic information as EI or CI+; however, some taxonomically significant compounds such as dipicolinic acid and poly(3-hydroxybutyrate) did produce negative ions. All ionization modes yielded spectra that could separate the bacteria by Gram-type when observed with principle components analysis (PCA). Chemical ionization in the positive ion mode produced the greatest amount of differentiation between the four genera of bacteria when the spectra where examined by PCA.

Bacteria↗

Photoelectron spectroscopy and DFT calculations of easily ionized quadruply bonded Mo2(4+) compounds and their bicyclic guanidinate precursors.

A series of five bicyclic guanidinate compounds containing various combinations of five- and six-membered rings and substituted alkyl groups have been shown by photoelectron spectroscopy to be easily ionized, with the one having two six-membered rings and four ethyl groups being the most easily ionized. The corresponding anions are capable of forming paddlewheel compounds having quadruply bonded Mo2(4+) units which are also easy to ionize. The most easily ionized compound is the ethyl-substituted Mo2(TEhpp)4 complex which has a broad first ionization band centered around 4.27 +/- 0.03 eV and an ionization onset at the very low energy of 3.93 +/- 0.03 eV. Even the compound with ligands containing two five-membered rings, which favors a long Mo-Mo separation because of the large ligand bite, has an ionization energy (4.78 eV) that is less than those of well-known organometallic reducing agents such as (eta5-C9Me7)2Co and (eta5-C5Me5)2Cr.

Journal Article↗

One- and two-photon ionization of DNA single and double helices studied by laser flash photolysis at 266 nm.

The ionization of the DNA single and double helices (dA)20, (dT)20, (dAdT)10(dAdT)10 and (dA)20(dT)20, induced by nanosecond pulses at 266 nm, is studied by time-resolved absorption spectroscopy. The variation of the hydrated electron concentration with the absorbed laser intensity shows that, in addition to two-photon ionization, one-photon ionization takes place for (dAdT)10(dAdT)10, (dA)20(dT)20 and (dA)20 but not for (dT)20. The spectra of all adenine-containing oligomers at the microsecond time-scale correspond to the adenine deprotonated radical formed in concentrations comparable to that of the hydrated electron. The quantum yield for one-photon ionization of the oligomers (ca. 10(-3)) is higher by at least 1 order of magnitude than that of dAMP, showing clearly that organization of the bases in single and double helices leads to an important lowering of the ionization potential. The propensity of (dAdT)10(dAdT)10, containing alternating adenine-thymine sequences, to undergo one-photon ionization is lower than that of (dA)20(dT)20 and (dA)20, containing adenine runs. Pairing of the (dA)20 with the complementary strand leads to a decrease of quantum yield for one photon ionization by about a factor of 2.

DNA↗

Intravenous magnesium sulphate relieves migraine attacks in patients with low serum ionized magnesium levels: a pilot study.

1. We tested the hypothesis that patients with an acute attack of migraine headache and low serum levels (< 0.54 mmol/l) of ionized magnesium are more likely to respond to an intravenous infusion of magnesium sulphate (MgSO4) than patients with higher serum ionized magnesium levels. 2. Serum ionized magnesium levels were drawn immediately before infusion of 1 g of MgSO4 in 40 consecutive patients with an acute migraine headache. 3. Pain reduction of 50% or more as measured on a headache intensity verbal scale of 1 to 10, occurred within 15 min of infusion in 35 patients. In 21 patients, at least this degree of improvement or complete relief persisted for 24h or more. Pain relief lasted at least 24h in 18 of 21 patients (86%) with serum ionized magnesium levels below 0.54 mmol/l, and in 3 of 19 patients (16%) with ionized magnesium levels at or above 0.54 mmol/l (P < 0.001). Mean ionized magnesium levels in patients with relief lasting for at least 24h were significantly lower than in patients with no relief or brief relief (P < 0.01). 4. Measurement of serum ionized magnesium levels may be useful in identifying patients with migraine headaches who may respond to an intravenous infusion of MgSO4.

Adult↗

Ionized and total serum magnesium concentrations in feline renal transplant recipients.

OBJECTIVE: To measure the blood concentrations of total and ionized serum magnesium in feline renal transplant recipients and to determine if there was a correlation between these concentrations and the development of neurological disorders after renal transplantation. STUDY DESIGN: Prospective clinical study. ANIMALS: Fourteen client-owned cats undergoing renal transplantation as a treatment for renal failure. Ten healthy adult cats were used to establish normal electrolyte concentrations. METHODS: Total and ionized serum magnesium as well as potassium and ionized calcium concentrations were measured in 14 renal transplant recipients at five intervals: preoperatively; immediately postoperatively; and 24, 48, and 120 hours postoperatively. The mean values from all 14 cats over each time interval were compared with the normal range. The serum concentration of these electrolytes, particularly magnesium, was evaluated in relation to the occurrence of neurological complications. RESULTS: Ninety-four percent of all ionized serum magnesium concentrations measured in clinical patients were below normal. Ninety percent of all total serum magnesium concentrations were within the normal range, and no cats had abnormally low total serum magnesium concentrations at any time. All clinical patients were hypocalcemic at all intervals. Sixty-six percent of all serum potassium concentrations were below normal. One cat in the study group experienced neurological problems, including seizures, in the immediate postoperative period. The signs appeared to be related to hypertension and responded to appropriate therapy. All electrolyte concentrations in this cat, including ionized magnesium, were within the same range of values as other clinical patients. CONCLUSIONS: Ionized serum magnesium concentrations are decreased in feline renal transplant recipients in the perioperative period; however, hypomagnesemia would not appear to be directly related to the development of neurological disorders. None of the study patients were hypomagnesemic when total serum magnesium concentrations were measured over the same intervals. In addition, ionized serum calcium concentrations and serum potassium concentrations are below normal in the perioperative period. CLINICAL SIGNIFICANCE: The specific clinical significance of these abnormalities is unknown. It is possible that the profound weakness and depression that is commonly seen in feline renal transplant recipients in the immediate postoperative period may be improved by supplementation with these electrolytes. Further work is needed to understand the implications of these abnormalities.

Animals↗