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PubMed1992

Formulation of radiographically detectable gastrointestinal contrast agents for magnetic resonance imaging: effects of a barium sulfate additive on MR contrast agent effectiveness.

Complete and homogeneous distribution of gastrointestinal (GI) contrast media are important factors for their effective use in computed tomography as well as in magnetic resonance (MR) imaging. A radiographic method (using fluoroscopy or spot films) could be effective for monitoring intestinal filling with GI contrast agents for MR imaging (GICMR), but it would require the addition of a radiopaque agent to most GICMR. This study was conducted to determine the minimum amount of barium additive necessary to be radiographically visible and to evaluate whether this additive influences the signal characteristics of the GICMR. A variety of barium sulfate preparations (3-12% wt/vol) were tested in dogs to determine the minimum quantity needed to make the administered agent visible during fluoroscopy and on abdominal radiographs. Solutions of 10 different potential GI contrast agents (Gd-DTPA, ferric ammonium citrate, Mn-DPDP, chromium-EDTA, gadolinium-oxalate, ferrite particles, water, mineral oil, lipid emulsion, and methylcellulose) were prepared without ("nondoped") and with ("doped") the barium sulfate additive. MR images of the solutions in tubes were obtained at 0.38 T using 10 different spin-echo pulse sequences. Region of interest (ROI) measurements of contrast agent signal intensity (SI) were made. In addition, for the paramagnetic contrast media, the longitudinal and transverse relaxivity (R1 and R2) were measured. A 6% wt/vol suspension of barium was the smallest concentration yielding adequate radiopacity in the GI tract. Except for gadolinium-oxalate, there was no statistically significant difference in SI for doped and non-doped solutions with most pulse sequences used. In addition, the doped and nondoped solutions yielded R1 and R2 values which were comparable. We conclude that barium sulfate 6% wt/vol added to MR contrast agents produces a suspension with sufficient radiodensity to be viewed radiographically, and it does not cause significant alteration in the MR signal appearance of most GICMR. These formulations can be useful for achieving optimal filling of the gastrointestinal tract prior to MRI.

Animals↗
PubMed1997

Additive Effects on the Microstructure Evolution in Hexadecyltrimethylammonium Bromide Solution and Its Rheological Properties

The effects of various additives on the microstructure evolution in hexadecyltrimethylammonium bromide micellar solution and its rheological properties are investigated. The additives considered in the present study are primary, secondary, and tertiary heptanols and sodium salicylate (NaSal). The microstructure is developed via the two-step shape transitions in the micellar solution; first, the initial spherical micelles undergo shape transition to rod-like or disc-like micelles as the micelles tend to be packed compactly with increase in the surfactant concentration; then further increment in the surfactant concentration makes the anisotropic rod-like micelles overlap each other. Solutions in these states exhibit typical non-Newtonian behaviors such as shear thinning at high shear rates. In the present study, the additive effects on the microstructure evolution are investigated by employing various techniques including a phase modulated flow birefringence, a dynamic light scattering and a dynamic oscillatory rheometry. The results show that addition of the solubilized additives enhances the microstructure transitions, which are affected by the additive concentration and its chemical structure. Presence of the cosurfactants such as heptanols with hydrophobic alkyl chains reduces the repulsion by forming surfactant-alcohol mixed micelles. The primary heptanol can penetrate easily into the micelles and aligned parallel to the surfactant molecules compared with the secondary and tertiary heptanols. Thus, the primary heptanol enhances the two-step shape evolutions more effectively than the other types. In the presence of NaSal, on the other hand, the micellar solution exhibits the viscoelastic properties and the yield stress owing to the formation of networked or worm-like micelles. Copyright 1997 Academic Press. Copyright 1997Academic Press

Journal Article↗
PubMed2000

Layer Buildup on Two-Layered Porous Substrate by Dip-Coating: Modeling and Effect of Additives on Growth Rate.

A model was developed to describe layer buildup on a two-layered substrate during dip-coating. In this model, the growth rate can be used to estimate the permeability of the cast layer. Layers of submicron alumina were dip-coated onto the asymmetric porous substrate to evaluate the model. The layer buildup and properties were studied with different slip additives. Slips were deflocculated with nitric acid or sodium poly(methacrylic acid) (Na-PMAA) to achieve electrostatic or electrosteric stabilization. Properties of the slips with electrostatic stabilization were varied with poly(vinyl alcohol) (PVA) concentration. Properties of the slips with electrosteric stabilization were varied with Na-PMAA concentration. PVA addition decreased the growth rate of the layer by reducing capillary pressure and increasing the viscosity of the filtrate. Electrostatic stabilization of the slip with PVA addition also led to almost complete deflocculation and a rather high density of layers. Na-PMAA addition led to fast growth of the layer. Na-PMAA addition increased the capillary pressure and had a minor effect on the viscosity of the liquid phase. Na-PMAA addition led to an incomplete dispersion of particles and therefore a high porosity of layers. Permeability values of coated layers obtained from the model were compared with permeability values predicted by the Kozeny-Carman equation. In the case of complete deflocculation of the slip, the permeability values estimated with the model correlated reasonably well with the predicted values. Incomplete deflocculation led to higher permeability values than predicted by the Kozeny-Carman equation. Copyright 2000 Academic Press.

Journal Article↗
PubMed2001

LiBr. 2H(2)O Crystallization Inhibition in the Presence of Additives.

Experiments have been performed to measure the effect of additives on the crystallization temperature of concentrated LiBr solutions cooled at a rate of 20 degrees C/h. The measured crystallization temperatures correspond not to the temperatures of equilibrium solubility but to the critical temperature for heterogeneous nucleation of the hydrated LiBr salt on the glass wall of the test tube containing the sample solution. Various additives at concentrations from 250 to 1500 ppm have been investigated. Some soluble additives further decreased the experimental crystallization temperature by as much as 13 degrees C, corresponding to 22 degrees C below the equilibrium solubility. Large decreases in the crystallization temperature can be correlated with large values of complexation constants of the additive for either the Li(+) or the Br(-) ion in solution. Solution complexation, however, is not sufficient to explain the magnitude of the decrease in the crystallization temperature. The only phenomenon capable of quantitatively explaining the magnitude of the decrease in the crystallization temperature is the change in the crystal/solution interfacial energy due to adsorption of the additive on the surface of the prenucleation embryos. A quantitative model of the crystal/solution interfacial energy due to adsorption has been developed using both the Langmuir and Gibbs adsorption equations, allowing the quantitative prediction of crystallization temperatures with additive concentration. Copyright 2001 Academic Press.

Journal Article↗
PubMed2002

Sonochemical synthesis of cerium oxide nanoparticles-effect of additives and quantum size effect.

Cerium oxide (CeO(2)) nanoparticles were prepared sonochemically, by using cerium nitrate and azodicarbonamide as starting materials, and ethylenediamine or tetraalkylammonium hydroxide as additives. The additives have a strong effect on the particle size and particle size distribution. CeO(2) nanoparticles with small particle size and narrow particle size distribution are obtained with the addition of additives; while highly agglomerated CeO(2) nanoparticles are obtained in the absence of additives. Monodispersed CeO(2) nanoparticles with a mean particle size of ca. 3.3 nm are obtained when tetramethylammonium hydroxide (TMAOH) is used as the additive and the molar ratio of cerium nitrate/azodicarbonamine/TMAOH is 1/1/1. Blue shifts of the absorption peak and the absorption edges of the products are observed in the UV-Vis absorption spectra as a result of the quantum size effect. The samples have been characterized using powder XRD, TEM, DLS, and absorption spectra.

Journal Article↗
PubMed1996

Enhanced biodegradation of diesel fuel through the addition of particulate organic carbon and inorganic nutrients in coastal marine waters.

Diesel fuel pollution in coastal waters, resulting from recreational boating and commercial shipping operations, is common and can adversely affect marine biota. The purpose of this study was to examine the effect of additions of particulate organic carbon (POC) in the form of naturally-occurring marsh grass (Spartina alterniflora), inorganic nutrients (nitrogen and phosphorus), inert particles, and dissolved organic carbon (DOC) on diesel fuel biodegradation and to attempt to formulate an effective bioremedial treatment for small diesel fuel spills in marine waters. Various combinations of treatments were added to water samples from a coastal marina to stimulate diesel fuel biodegradation. Diesel fuel was added in concentrations approximating those found in a spill and biodegradation of straight chain aliphatic constituents was estimated by measuring mineralization of 14C hexadecane added to diesel fuel. All treatments that included POC showed stimulation of biodegradation. However, the addition of inert particles (glass fiber filters and nylon screening) caused no stimulation of biodegradation. The addition of nitrogen and phosphorus alone did not result in stimulation of biodegradation, but nitrogen and Spartina (although not phosphorus and Spartina) did result in stimulation above that of Spartina alone. Maximum biodegradation rates were obtained by the addition of the Spartina POC, ammonium, and phosphate. The addition of mannitol, a labile DOC source with POC and phosphate resulted in a decrease in diesel fuel biodegradation as compared to POC and phosphate alone. The seasonal pattern of diesel fuel biodegradation showed a maximum in the summer and a minimum in the winter. Therefore, of the treatments tested, the most effective for bioremediation of diesel fuel in marine waters is the addition of POC, nitrogen, and phosphorus.

Biodegradation, Environmental↗
PubMed2004

Breast cancer in patients with residual invasive carcinoma is more accurately staged with additive tumor size assessment.

BACKGROUND: Accurate assessment of tumor size for patients with breast cancer undergoing re-excision following breast-conserving therapy is important for appropriate staging and adjuvant treatment. We investigated the accuracy of additive vs. nonadditive size assessment in determining final tumor stage. METHODS: Patients with infiltrating carcinoma in the initial excision and in at least one additional re-excision (re-excision positive; n = 89) had tumor size assessed with additive and nonadditive techniques. This group was compared with patients undergoing re-excision but without identifiable residual carcinoma (re-excision negative; n = 105) regarding rates of lymph node (LN) metastasis. RESULTS: The re-excision positive patients had a different median final tumor size depending on the size assessment technique used (nonadditive: 1.8 cm; additive: 3.0 cm; P <.0001). Both groups of patients had a median tumor size consistent with T1c staging in nonadditive size assessment. However, re-excision positive patients had a significantly higher incidence of LN metastasis (P <.05) than did re-excision negative patients. Both groups were then separated into T1 and T2 stages and the LN metastasis rates were assessed. Compared with nonadditive size assessment, additive size assessment distributed re-excision positive patients into T stages whereby the LN metastasis rates more closely approximated those of re-excision negative patients (T1, 3% vs. 6% difference; T2, 4% vs. 13% difference). CONCLUSIONS: With regard to LN metastasis, staging for patients with residual invasive carcinoma in re-excision specimens is more accurate with additive tumor size assessment.

Breast Neoplasms↗
PubMed2001

Feasibility and potential gains of enhancing the subacute rat study protocol (OECD test guideline no. 407) by additional parameters selected to determine endocrine modulation. A pre-validation study to determine endocrine-mediated effects of the antiandrogenic drug flutamide.

Groups of five male and five female Wistar rats were treated by gavage with 0, 1, 10, and 100 mg flutamide/kg body weight for at least 28 days to investigate whether proposed enhancements to the current subacute rodent OECD test guideline no. 407 could be included into the testing routine, which of the current and/or additional parameters would detect endocrine-mediated effects of flutamide reliably and sensitively, and to provide information on intra-laboratory variability. Two identical studies were performed concurrently. The enhanced protocol requests the additional determination of the specific hormones triiodothyronine, thyroxine, thyroid stimulating hormone, follicle stimulating hormone (FSH), luteinizing hormone (LH), estradiol, prolactin, testosterone, corticosterone; of oestrus cyclicity and necropsy of all females in the dioestrus stage; of the number of homogenization-resistant testicular spermatids and the number, motility, viability, and morphology of cauda epididymal spermatozoa; of additional organ weights (pituitary, ovaries, uterus, thyroid, male accessory reproductive organs); and of the histopathology of additional organs (pituitary, epididymides, coagulation glands, pancreas, vagina). From a technical standpoint, it was possible to conduct a study according to the enhanced protocol, however, with substantial additional effort, an increase in costs by some 67%, and logistic problems. In line with the specific pharmacological effect of flutamide, treatment-related changes were mainly found in male rats, while females were hardly affected by 100 mg/kg. In male rats, 100 mg/kg was the maximal tolerated dose resulting in reduced body weight gain, but no or little other effects on clinical, haematological, clinico-chemical, or behavioral parameters, and 1 mg/kg was the no-observed-adverse-effect level. Antagonism of peripheral androgen receptors by flutamide resulted in decreased relative organ weights of male accessory reproductive organs, changes that were reliably detected in both studies at 100 mg/kg, but only in one of both studies at 10 mg/kg. Corresponding histopathological changes were also detected reliably at 100 mg/kg. Antagonism of central androgen receptors by flutamide increased LH and FSH levels. LH stimulation of testicular Leydig cells in turn increased testosterone and estradiol levels. Again, all these changes were detected reliably at 100 mg/kg, but only in one of both studies at 10 mg/kg. Corresponding histopathological alterations (increase of LH- and FSH-secreting cells, Leydig cell hypertrophy) were detected reliably and sensitively at 10 mg/kg. Studies on liver enzymes performed outside the scope of the enhanced protocol showed that flutamide at 100 mg/kg generally induced hepatic enzyme activities, but decreased the activity of the sex-specific testosterone-dependent liver enzyme CYP2C11 in male rats. The laboratory methods employed yielded reliable results, i.e., 93.6% of the quantitative measurements obtained in both studies were in agreement. Doubling the animal number from five to ten per sex and dose does not increase the sensitivity of detection of endocrine-mediated effects above the level already provided by histopathological examination of groups of five animals. Some of the proposed enhancements evaluated (additional organgravimetry and histopathology) were helpful in detecting the endocrine-mediated effects of flutamide reliably, while others did not contribute towards this aim (spermatology resulted in doubtful effects, female cyclicity was not affected, hormone determinations provided mechanistic information). Ongoing testing according to the revised version of the enhanced OECD test guideline no. 407 protocol and using ten compounds interfering with the endocrine system by different mechanisms will result in the identification of the most appropriate enhancements.

Administration, Oral↗
PubMed2006

Effect of addition of antibiotics and an antioxidant on the stability of tissue reference materials for domoic acid, the amnesic shellfish poison.

Five separate reference materials (RMs) were prepared from a mussel (Mytilus edulis) tissue containing domoic acid (DA) from scallop hepatopancreas (Pecten maximus). Homogenates were separately spiked with antibiotics, an antioxidant, or a combination of both. Control materials did not contain any additives and were prepared from lightly cooked and autoclaved mussel tissues. Stability studies were run over a 148-day period at three different temperature conditions: -20 degrees C, +4 degrees C and +40 degrees C. DA contents in all materials were characterised by HPLC-UV. Homogeneities were demonstrated at the beginning of the study, with coefficients of variance of less than 4% (n = 9). DA was stable at -20 degrees C in all materials. The control materials showed significant degradation after two days at +40 degrees C, and after eight days at +4 degrees C. Each of the materials containing additives demonstrated better stability during the initial period of the study. In addition there was no significant degradation in any of the materials with additives stored at +4 degrees C over the duration of the study. The material containing a combination of the antibiotics and the antioxidant displayed the best stability of all the materials. There was no significant reduction in DA concentration at all temperature conditions after eight days, and after 32 days the decrease at +40 degrees C was still <20 %. Following this, a DA laboratory reference material (LRM) was prepared and, based on previous results, spiked with both the antioxidant and antibiotics. A short-term stability study on this material gave similar results to the corresponding material in the additives study. This study shows that combined use of the additives investigated in the preparation of a mussel tissue reference material for DA ensures analyte stability for a period of up to eight days at temperatures of up to +40 degrees C, a condition that is particularly important when shipping test materials globally. Aliquots of individual feasibility materials used in the study.

Animals↗
PubMed2004

Chemosensory additivity in trigeminal chemoreception as reflected by detection of mixtures.

A series of experiments probed into the degree of chemosensory detection additivity exhibited by mixtures of ethyl propanoate and heptanoate in terms of their trigeminal detectability via nasal pungency (i.e., irritation) and eye irritation. Nasal pungency was tested in subjects lacking a functional sense of smell (i.e., anosmics) to avoid olfactory biases. First, we built concentration-detection functions for each chemical and sensory endpoint. Second, we used the data from the functions to prepare mixtures of the two compounds in complementary proportions, and suitable single-chemical standards, all of which should be equally detectable under a rule of complete additivity, i.e., independence of detection. Third, we compared the experimentally obtained detectability with that expected under such rule. The outcome revealed that, at a low detectability level (but still above chance), the mixtures showed complete additivity for both trigeminal endpoints. At a high detectability level (but below perfect detection), the mixtures showed complete additivity for nasal pungency but less than complete additivity for eye irritation. In the context of previous studies, the results consolidate a picture of higher degree of detection additivity at perithreshold levels in trigeminal than in olfactory chemoreception. The outcome presents another line of evidence suggesting broader chemical tuning in chemesthesis compared to olfaction.

Adult↗
PubMed2002

Impact of additional pleurodesis in video-assisted thoracoscopic bullectomy for primary spontaneous pneumothorax.

BACKGROUND: Bullectomy for primary spontaneous pneumothorax has been associated with high postoperative recurrence rates when video-assisted thoracoscopic surgery (VATS) has been used rather than thoracotomy. The aim of this study was to evaluate the efficacy and identify the disadvantages, if any, of adding pleurodesis to VATS bullectomy to prevent recurrent pneumothorax. METHODS: Fifty-three patients who underwent VATS bullectomy with additional pleurodesis for pneumothorax after November 1996 and 50 who underwent VATS bullectomy alone before October 1996 were compared retrospectively in terms of intraoperative factors and postoperative chest pain, pulmonary function, and pneumothorax recurrent rates. Pleurodesis was achieved by electrocauterizing the upper surface of the parietal pleura in a patchy fashion. RESULTS: There were no significant differences between the additional pleurodesis group and the bullectomy alone group in terms of age, sex, operating time, intraoperative bleeding, number of resected bullae, duration of chest drainage, or volume of fluid drained. Postoperative chest pain and pulmonary function were also similar in both groups. A recurrent pneumothorax occurred in one patient (1.9%) in the additional pleurodesis group; this recurrence rate was significantly lower than that for the bullectomy alone group (eight patients, 16%; p = 0.029). Although the mean postoperative follow-up period was considerably shorter in the additional pleurodesis group (38 months [range, 26-49]) than in the bullectomy alone group (63 months [range, 50-72] ), eight (89%) of all nine recurrences occurred within 26 months of surgery-i.e., within the minimum follow-up period for the additional pleurodesis group. CONCLUSIONS: Pleurodesis is a minimally invasive technique that is effective in preventing postoperative recurrences of pneumothorax when added to VATS bullectomy. Additional pleurodesis has no disadvantages vs bullectomy alone in terms of worsening postoperative chest pain or pulmonary function.

Adult↗
PubMed2004

Effect of heparin addition on expansion of cord blood hematopoietic progenitor cells in three-dimensional coculture with stromal cells in nonwoven fabrics.

Primary human cord blood mononuclear cells (CB MNCs) were inoculated into layers of primary human bone marrow stromal cells prepared in a nonwoven fabric porous carrier [three dimensional (3-D)] or on a dish [two dimensional (2-D)] using a cytokine-free medium and were cultured for 7 days with or without the addition of heparin. The number of progenitor cells increased threefold during the 3-D coculture, whereas it decreased in the 2-D culture. Heparin addition to the 3-D coculture further increased the number of progenitors twofold, whereas the addition of desulfated heparin had no effect. The heparin effect was also observed in a 3-D culture of CB MNCs without stromal cells when conditioned medium was employed. The coating of the carrier with N-(O-beta-(6-O-sulfogalactopyranosyl)-6-oxyhexyl)-3,5-bis (dodecyloxy)-benzamide instead of heparin addition also increased the number of progenitor cells in the 3-D culture of CB MNCs without stromal cells when the conditioned medium was employed. The 3-D coculture constructed with nonwoven fabrics and stromal cells was clearly superior to the 2-D culture because of the expansion of CB hematopoietic progenitor cells without cytokine addition. Heparin addition to the 3-D coculture further increased the number of progenitor cells, which may result from a synergistic effect of soluble cytokines produced by stromal cells with the sulfur group of heparin.

Animals↗
PubMed1995

Overlapping stage models and reaction time additivity: effects of the activation equation.

Many experimental factors have been found to affect mean reaction time (RT) additively in factorial experiments. What sorts of RT models are compatible with this fact? Sternberg (1969) showed that serial, discrete-stage models are consistent with additivity, and as a result additivity has sometimes been regarded as evidence in favor of such models. However, McClelland (1979) showed that an alternative "cascade" model, which violates crucial assumptions of discrete-stage models, also predicts RT additivity in many cases. This article examines various modified versions of the cascade model, and shows that mean RT additivity arises from many overlapping stage models, including some with thresholds, information quantization, and nonlinear activation transformations. This suggests that other aspects of the data besides mean RT additivity should be examined to distinguish between serial and overlapping stage models.

Arousal↗
PubMed1986

Accuracy of addition silicones as a function of technique.

Both addition and condensation silicones produced stone dies that were larger in diameter and shorter in height than the tooth preparation. For both materials, there was little change in the distance between stone die preparations compared with the standard. The most significant difference between types of silicone was that condensation silicones produced significantly shorter dies (-0.24% to -0.37%) than addition silicones (-0.08%). Among addition silicones, Cinch produced more than twice as much vertical change (-0.16%) than the other three products (-0.06%). Since castings made from a short die will not seat completely on the prepared tooth, these results support the use of three of the four addition silicones tested. The same accuracy of impressions was achieved for all techniques (that is, putty/wash, single mix, and double mix) when addition silicones were used, whereas the putty/wash technique produced the most accurate dies for the condensation silicones. The custom tray produced dies that were much more accurate in vertical dimensions (-0.03%) than the stock trays (-0.15% to -0.21%). Thus the custom tray is the impression tray of choice, even for addition silicones, which produce relatively little polymerization shrinkage and are dimensionally stable.

Chemical Phenomena↗
PubMed1989

Temporal induction of blackness--II. Spectral efficiency and tests of additivity.

The perception of blackness was investigated by measuring spectral efficiency and field additivity under conditions of temporal induction. For both purposes, observers foveally viewed a 0.75 deg, inducing stimulus for 5 sec followed immediately (or 200 msec later) by a broadband (5500 K) reference stimulus of the same size and spatial location. For spectral efficiency measurements, the inducing field was a monochromatic light between 400 and 700 nm (10 nm steps), while the additivity studies involved various wavelength mixtures. The psychophysical task was to increase the radiance of the inducing field until the reference stimulus just turned completely black. For two observers the spectral efficiency function of temporally-induced blackness more closely resembled their heterochromatic flicker photometry (HFP) function than their direct brightness-matching function. Their brightness functions were characterized by an inflection at about 580 nm which is generally ascribed to subtractive cone interactions, but their blackness-induction and HFP functions did not show this inflection. The brightness function of the third observer did not show an inflection at 580 nm, thereby making it difficult to differentiate between her three spectral efficiency functions. Overall, the subtle differences between the various spectral efficiency functions made it difficult to determine whether blackness induction was more similar to HFP or brightness matching. The results from the additivity tests of blackness induction, HFP and direct brightness matching removed this ambiguity from the spectral efficiency findings. Blackness induction and HFP were shown to be additive, whereas the results from brightness matching showed clear additivity failures of the cancellation type. These data support the view that the perception of blackness is mediated by neural mechanisms that additively combine the inputs of middle- and long-wave photoreceptors.

Adult↗
PubMed1994

Partial additivity of rod signals with M- and L-cone signals in increment detection.

Test additivity experiments revealed the combination rules for increment detection by rods and either M- or L-cone-dominated mechanisms isolated by means of chromatic adaptation (Stiles' pi 4 and pi 5, respectively). Increment thresholds were measured for single test wavelengths detected by each mechanism. Pairs of test wavelengths were then superimposed, and increment thresholds were measured for simultaneous detection by both rod and cone mechanisms. The observed degree of additivity was corrected (reduced) to compensate for the partial detection by each mechanism of both test wavelengths in the combined stimuli. We find that subthreshold rod signals are partially additive with subthreshold signals from both M- and L-cones. The degree of additivity is high and similar for both M- and L-cones: less than the ideal prediction of linear addition, but greater than that predicted by either probability summation of independent mechanisms or orthogonal vector addition.

Color Perception↗
PubMed1984

Requirement of a downstream sequence for generation of a poly(A) addition site.

The 3' terminus of most, if not all, eucaryotic polyadenylated mRNAs is formed as a result of endonucleolytic cleavage of a larger precursor RNA. That is, transcription does not terminate at the mRNA 3' sequence but rather proceeds through this site, terminating at some distance downstream. Using a plasmid containing the adenovirus E2A transcriptional unit, we have investigated the sequence requirement for the formation of a mature mRNA 3' terminus, focusing on the role of sequences immediately distal to the poly(A) addition site. Deletion mutants were constructed in the region distal to the poly(A) addition site and assayed by transfection into human 293 cells. The results demonstrate that 35 nucleotides distal to the site of poly(A) addition are sufficient for the formation of a mature E2 mRNA. However, removal of an additional 15 nucleotides, leaving 20 nucleotides distal to the poly(A) site, abolished the ability to produce functional E2A mRNA. The defect in the production of functional mRNA from such a mutant appears to be in the proper cleavage of the primary transcript at the poly(A) addition site. It would thus appear that sequences immediately distal to the site of poly(A) addition do not contribute to the mature mRNA but are essential for the formation of mature mRNA.

Adenoviruses, Human↗
PubMed1983

Genotoxicity studies with a blend of zinc dialkyldithiophosphate lubricant additives.

The mutagenic activity of a blend of primary zinc dialkyldithiophosphate lubricant additives suspended in process oils and a blend of the process oils alone was investigated in agar layer cultures of Salmonella typhimurium TA1535, TA1537, TA1538, TA98 and TA100, both with and without the incorporation of a rat liver microsomal activation system (S9). Zinc dimethyldithiocarbamate was tested as a structurally-related model bacterial mutagen, and, in additional control experiments, the mutagenic activity of zinc dimethyldithiocarbamate and benzo[a]pyrene was investigated in combination with the blend of process oils in selected bacterial tester strains. Transformation frequencies of BHK cells were determined by colony growth in soft agar culture following treatment with a blend of the zinc dialkyldithiophosphate lubricant additives suspended in process oils, a blend of the process oils, 7,12-dimethylbenzanthracene, both in the presence and in the absence of a blend of the process oils, or zinc dimethyldithiocarbamate. All experiments incorporated a rat liver microsomal activation system (S9). Application of a blend of the zinc dialkyldithiophosphate additives or a blend of the process oils used in their manufacture did not increase the reverse mutation frequencies of Salmonella typhimurium TA1535, TA1537, TA1538, TA98 or TA100, or significantly increase the transformation frequency of BHK cells under the experimental conditions described. Zinc dimethyldithiocarbamate increased the reverse mutation frequency in some bacterial tester strains, but did not significantly increase the transformation frequency of BHK cells under the described experimental conditions. The addition of the blend of the process oils in combination with the control materials, zinc dimethyldithiocarbamate or benzo[a]pyrene had an inhibitory effect on the mutagenic activity at high doses of each in the bacterial assays, and in the BHK assay the transforming ability of 7,12-dimethylbenzanthracene was suppressed in the presence of the blend of the process oils. Thus, the additive materials showed no evidence of genotoxic activity in the bacterial mutation assays, or in the BHK transformation assay under the experimental conditions described.

Animals↗