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Qualitative and quantitative methods for analysis of 2-methyl-3(2-pyridyl-methyl)-3H-pyrido-[2,3-d]-pyrimidin-4-on dihydrochloride were developed. Qualitative analysis included characteristic reactions, chromatographic investigations, interpretation of IR and UV spectra and quantitative analysis included spectrophotometric, argentometric and alkalimetric methods.
A comparison was made of cerebellar dendritic development in the normal rat and in a new model of phenylketonuria, the phenylacetate-treated suckling rat. Golgi stain analysis of the Purkinje cells shows striking regional variations in the dendritic growth. These variations were observed in both the control and phenylacetate-treated animals and were especially striking before 15 days of life. Quantitative analysis of the dendritic tree revealed, in the phenylacetate-treated rat, a significant reduction in the total number of dendritic branches. However, the individual terminal dendritic length was largely unaltered. These effects of phenylacetate differ from those of deafferentation, and starvation. Results of this investigation clearly define the harmful effects of phenylacetate on developing neurons and are compatible with the clinical observation that brain damage in phenylketonuria occurs mainly during the first few years of life, the critical period of neuronal development.
The reactions of the cations with 2-mercapto-5-R-amino-1,3,4-thiadiazole derivatives were studied conductometrically with the purpose of establishing a new conductometric method for the quantitative analysis of Pb(II) and Cd(II). Aqueous solutions of Pb(NO3)2 and Cd(CH3COO)2 were titrated with hidroalcoholic solutions of 2-mercapto-5-amino-1,3,4-thiadiazole (MATD), 2-mercapto-5-allylamino-1,3,4-thiadiazole (MA1ATD) and 2-mercapto-5-acetylamino-1,3,4-thiadiazole (MAcATD) and 2-mercapto-5-phenilamino-1,3,4-thiadiazole (MFATD) in different concentrations. The reactions takes place at pH 6.5 (realised with acetate buffer). A linear classical titration curves was obtained. In solutions more concentrated than 10(-2) M just one equivalence point can be noticed, corresponding to 1:2 Me:R stoechiometries. For concentration less than 10(-2) M two equivalence point were observed at 1:1 and 2:1 ratio of Me:R, indicating the step formation of the complex. Accurate conductometric determinations can be made using the second break points of the titration curves as equivalence points. The amounts of Cd(II) and Pb(II) taken and recovered are good, with an error less than 1%.
The analysis of acylglycines is an important biochemical tool for the diagnosis of inherited disorders of mitochondrial fatty acid beta-oxidation. A stable isotope dilution gas chromatography negative chemical ionisation mass spectrometry method for the quantitative analysis of short- and medium-chain acylglycines as their bis(trifluoromethyl)benzyl (BTFMB) ester derivatives is described. The diagnostic usefulness of the method was demonstrated in nine patients with medium-chain acyl-coenzyme A (CoA) dehydrogenase (MCAD) deficiency, and seven patients with multiple acyl-CoA dehydrogenation defect (MAD). The urinary acylglycine profiles in these patients were compared to those in controls (n = 19), children on a medium-chain triglyceride (MCT) supplemented diet (n = 4), and patients with various other diseases (n = 5).
BACKGROUND AND AIMS: Plant architecture and its interaction with agronomic practices and environmental constraints are determinants of the structure of the canopy, which is involved in carbon acquisition and fruit quality development. A framework for the quantitative analysis of grapevine (Vitis vinifera) shoot architecture, based on a set of topological and geometrical parameters, was developed for the identification of differences between cultivars and the origins of phenotypic variability. METHODS: Two commercial cultivars ('Grenache N', 'Syrah') with different shoot architectures were grown in pots, in well-irrigated conditions. Shoot topology was analysed, using a hidden semi-Markov chain and variable-order Markov chains to identify deviations from the normal pattern of succession of phytomer types (P0-P1-P2), together with kinematic analysis of shoot axis development. Shoot geometry was characterized by final internode and individual leaf area measurements. KEY RESULTS: Shoot architecture differed significantly between cultivars. Secondary leaf area and axis length were greater for 'Syrah'. Secondary leaf area distribution along the main axis also differed between cultivars, with secondary leaves preferentially located towards the basal part of the shoot in 'Syrah'. The main factors leading to differences in leaf area between the cultivars were: (a) slight differences in main shoot structure, with the supplementary P0 phytomer on the lower part of the shoot in 'Grenache N', which bears a short branch; and (b) an higher rate and duration of development of branches bearing by P1-P2 phytomers related to P0 ones at the bottom of the shoot in 'Syrah'. Differences in axis length were accounted for principally by differences in individual internode morphology, with 'Syrah' having significantly longer internodes. This trait, together with a smaller shoot diameter, may account for the characteristic 'droopy' habit of 'Syrah' shoots. CONCLUSIONS: This study highlights the architectural parameters involved in the phenotypic variability of shoot architecture in two grapevine cultivars. Differences in primary shoot structure and in branch development potential accounted for the main differences in leaf area distribution between the two cultivars. By contrast, shoot shape seemed to be controlled by differences in axis length due principally to differences in internode length.
Insurance medical consultation by a medical consultant or medical director is well established at many companies in the German private health insurance sector. Nevertheless, the field of activity of the medical consultant is hardly known to the public and even less scientifically investigated. The present study deals with a quantitative analysis of insurance medical enquiries to medical consultants. For this, the total of all insurance medical enquiries in a whole year have been checked and classified. The total of 5116 enquiries shows that a large demand for consultation refers to the medical necessity of medicaments, followed by consultation on complementary and alternative medicine (CAM). Further important fields of consultation are the assessment of out-patient and in-patient psychotherapy, the verification of the extent of physical measures, the duration of hospital treatments, the extent of laboratory testing, the examination of new medical methods and the definition of cure measures in comparison with hospital treatment. Increasing demand exists for the definition of the necessity of out-patient and in-patient operations, the definition of cosmetic surgery and medically necessary plastic surgery and for questions of reproductive medicine. There is also increasing demand for consultation regarding lifestyle drugs and anti-aging medicine.
Electron paramagnetic resonance (EPR) spectroscopy was used to study the radical species formed during the oxidation of an active pharmaceutical ingredient in the solid state. It was found that the extent of radical generation correlated to the formation of an oxidative degradation product. Multifrequency EPR and electron nuclear double resonance spectroscopy gave additional information on the identity of the organic radical species involved in the oxidation process, and a mechanism was proposed for the degradation, involving the formation of both carbon-centered and peroxy radicals. The multivariate analysis technique of partial least-squares (PLS) regression was then used to determine the extent of oxidation of the active pharmaceutical ingredient from the EPR spectra. The suitability of this approach was demonstrated from its application to a series of standards. The conventional approach for the quantitative analysis of EPR spectra is to measure the peak height or to perform double integration of the spectral region containing the signal of interest. Both of these methods have intrinsic errors associated with them, particularly for weak EPR signals with a poor signal-to-noise ratio or a sloping background response. The results obtained showed that greatly improved quantitation was obtained using the PLS regression approach.
Prior studies from our institution have shown that single photon emission computed tomography is sensitive (100%) in predicting patients at risk for serious arrhythmias. However, the positive predictive value is low (15% to 20%). The purpose of this study was to determine if quantitative analysis of single photon emission computed tomographic defects could improve predictive value. One hundred seventy-five patients with positive single photon emission computed tomographic scans were studied. One hundred two patients developed arrhythmias, 42 of which were ventricular. Arrhythmias were associated with all defect loci and all defect sizes. The incidence of arrhythmias did increase with increasing size. Patients were at risk for arrhythmias up to 72 hours after trauma. The value of single photon emission computed tomography is its ability to predict patients at risk for arrhythmias. This study shows that any single photon emission computed tomographic defect, regardless of location or size, is a significant predictor of arrhythmias.
In the absence of a direct and specific marker for basket cells, the aim of this paper was to use available data to estimate the density of basket cell synaptic input to smooth and spiny neurons within layer IV of cat striate visual cortex (area 17). A linear quantitative analysis of layer IV basket cell connectivity data suggests that on average basket cells (1) comprise 25-35% of all GABAergic neurons in layer IV (3552-4736 cells mm(-3)), (2) account for 30-41% of all putative inhibitory dendritic synapses of layer IV spiny stellate cells (145-195 synapses cell(-1)) and a similar proportion of layer IV basket cells (25-37%, 71-107 synapses cell(-1)), and (3) provide each layer IV spiny cell with 13-45 axons and each layer IV basket cell with 6-29 axons. These estimates suggest that basket cells may be less common and provide a smaller proportion of the dendritic synaptic input to layer IV spiny and smooth neurons than previously thought. In addition, the analysis indicates that a layer IV spiny stellate cell may receive on average as many synapses and axons from layer IV basket cells as from lateral geniculate relay cells. Based on this potential numerical similarity, a geniculate-basket synaptic pairing in a spine-shaft microcircuit is hypothesized. This microcircuit could implement a type of local (dendritic) push-pull interaction underlying subfield antagonism.
BACKGROUND: Sodium polystyrene sulfonate (Kayexalate) commonly is used in treating hyperkalemia. As a cation exchange resin, it also can be used to reduce the potassium content of enteral nutrition formulas. This study evaluates the use of Kayexalate to reduce potassium in one high-protein enteral formula and describes the quantitative analysis of the product. METHODS: Sodium polystyrene sulfonate and enteral formula were mixed into a slurry and allowed to settle, and then the supernatant was decanted off and tested as samples. Three sample concentrations were analyzed: a control not subjected to potassium reduction, 0.5 g of Kayexalate per mEq K+ sample, and a 1 g/mEq K+ sample. Of these samples, moisture, lipid, protein, carbohydrate, ash, and mineral content were obtained. RESULTS: Compared with the control, the percentage decrease of potassium ranged from 25% to 36%, depending on the concentration of Kayexalate. A significant increase of 324% in sodium concentration was found in the 1.0 g/mEq K+ sample. Although there was no change in magnesium content, a slight increase in phosphorus, iron, and zinc was evident. CONCLUSIONS: The treatment of an enteral formula with sodium polystyrene sulfonate significantly increases its sodium content, with a modest decrease in potassium content. Clinicians using this method in clinical practice should be aware of the increase in sodium content.
Continuous replacement of sensory neurons is a normal process in the olfactory system of adult vertebrates. The capacity for replacement following experimentally induced degeneration of sensory neurons makes this system ideal for the study of the dynamics of neural populations. A quantitative analysis was made to determine the time course and degree of cell replacement in the olfactory epithelium following unilateral bulbectomy in adult hamsters. Histological measurements of number of cells and epithelial thickness were made for up to 194 days postoperatively. Results for each experimental animal were expressed as a percentage of the contralateral control side. There was an immediate degeneration of cells, the number decreasing to 39% by day 4. During days 4--15 new growth resulted in an increase in cell number, which was maintained at a level of 60--70% through day 194. Epithelial thickness decreased to 60--70% during the degeneration period, but there was no recovery during subsequent days 4--194. Analysis of epithelial cells by cell type (supporting, receptor, and basal cells) showed that changes in cell numbers were limited to the neural cell populations (receptor and basal). This study confirms that olfactory sensory neurons are capable of replacement following degeneration in spite of the absence of normal target tissue. However, the observed recovery does not reach control levels and the functional capacity of replacement neurons requires further study.
We have demonstrated that myocardial mitochondrial DNA (mtDNA) with a 7,436 base-pair deletion existed in all subjects that were over 70 years old. Since each mitochondrion has two or three copies of its own DNA, quantitative analysis is required for the evaluation of the role of mtDNA with deletions in the age-related deterioration of cardiac performance. For this purpose, the kinetic polymerase chain reaction (PCR) method developed in our laboratory was used in this investigation to determine myocardial mtDNA with this 7,436 base-pair deletion in human cadavers of various ages. The mtDNA population with this deletion increased exponentially with age [log f (% of deleted mtDNA) = -3.136 + 0.0454 x age, r = 0.95, P less than 0.01)], and was estimated at 3% and 9% in subjects of age 80 and 90, respectively. The deleted portion encodes 7 subunits of the mitochondrial ATP production system, and a population of mtDNA with this deletion over a certain threshold might induce a significant deterioration of cardiac energy metabolism. Cardiac function is known to deteriorate with age, and an increase in the population of mtDNA with deletion is likely to be an important contributing factor to aged heart (presbycardia).
We have measured the excitation spectrum of six micron magnetic squares using x-ray magnetic circular dichroism. We observe all three excitations expected in a Landau flux-closure pattern. High temporal and spatial resolution allows quantitative analysis of the excitations. A short magnetic in plane pulse excites the magnetic element and we observe precessional motion of the magnetization within the domains as well as a domain wall mode and vortex motion. The vortex moves perpendicular to the excitation field and relaxes without showing a circulating orbit.
Higher plant plastid genomes encode rRNAs, tRNAs, and protein subunits of the RNA polymerase, ribosomes, and the photosynthetic apparatus which vary over 1000-fold in abundance. Quantitative analysis of transcription and RNA levels was carried out on 15 plastid genes which are located in 14 different transcription units covering 50% of the barley plastid genome. Transcription of 16S rRNA, trnfM-trnG, and trnK was high relative to most other plastid genes. Transcription of trnfM-trnG was 5 times greater than trnK indicating that differences in tRNA levels in plastids could be due, in part, to differences in transcription. Among the protein coding genes, mRNA levels varied over 900-fold and transcription over 300-fold. The gene showing the lowest transcription rate and mRNA level, rpoB, is located in a gene cluster which encodes subunits of the plastid RNA polymerase (rpoB-rpoC1-rpoC2). RpoA, which encodes the alpha subunit of the RNA polymerase, was located in a gene cluster encoding ribosomal proteins (rpl23, rps19, rpl16) and infA. RNA from this gene cluster is 30-fold more abundant than rpoB mRNA, suggesting that expression of rpoA is regulated at the level of translation or protein stability. Polycistronic operons encoding subunits of the photosynthetic apparatus (psbB-psbH-petB-petD; psbK-psbI-psbD-psbC; atpB-atpE; psaA-psaB) had higher transcription rates and correspondingly higher mRNA levels than genes which encode ribosomal proteins or RNA polymerase subunits. RbcL and psbA, which are located in separate transcription units, exhibited the highest transcription rates and mRNA levels. Correspondence between transcription rate, mRNA level, and protein abundance indicates that transcription is a primary determinant of barley plastid gene expression. In addition, a 30-fold variation in predicted mRNA stability was observed which further increases the dynamic range of plastid mRNA abundance.
In a study of the etiological factors in esophageal cancer, Roussin red methyl ester was isolated and identified in pickled vegetables of Linxian County, North China, where there is a high incidence of esophageal cancer. In this paper, a method is described for quantitative analysis of Roussin red methyl ester in pickled vegetables by gas chromatography-high-resolution mass spectrometry single-ion monitoring. The content of Roussin red methyl ester in pickled vegetables from Linxian has been found to be 0.1 to 4.5 mg/kg, and that from Beijing is below 0.005 mg/kg, which is the detection limit of the analytical method used. The marked difference between the contents might be one of the possible reasons for the difference in esophageal cancer incidence between the two regions.
Telomerase activity has been associated with almost 90% of malignant human cancers from a variety of tissue sources, making it one of the most prominent molecular cancer markers known to date. As such, telomerase has become a very attractive diagnostic and therapeutic target. The advent of the telomeric repeat amplification protocol (TRAP) has allowed for the semiquantitative detection of telomerase from limiting sample amounts. Both the standard TRAP assay and a real-time assay using Amplifluor technology with primers designed specifically for telomerase activity amplification were used to quantitatively assess telomerase activity in primary tumors and tumor-derived cell lines. We have adapted the recently developed TRAPeze XL telomerase detection kit (Intergen, Gaithersburg, MD) for use with real-time polymerase chain reaction for more accurate quantification and high-throughput capabilities. In doing so, the reliability, assay time, and accuracy of quantitation have all been dramatically improved. A comparison of the quantitative analysis for the standard TRAP assay versus the real-time assay using 19 breast tumors revealed telomerase quantitation and standardization using the real-time assay was superior to the standard assay. Our data suggest that this assay will be useful for clinical and research studies involving detection of telomerase activity as it relates to cancer diagnosis.
Infantile hydronephrosis detected by ultrasonography poses a clinical dilemma on how to treat the condition. This article reports a retrospective study to evaluate infantile hydronephrosis due to suspected ureteropelvic junction (UPJ) obstruction by means of standardized diuretic renography and to speculate its usefulness for quantitative assessment and management of this condition. Between November 1992 and July 1999, 43 patients who had the disease detected in their fetal or infantile period were submitted to this study. Standardized diuretic renograms were obtained with 99mTc-labeled diethylene-triaminepenta-acetate (Tc-99m-DTPA) or 99mTc-labeled mercaptoacetyl triglycine (Tc-99m-MAG3) as radiopharmaceuticals. Drainage half-time clearance (T 1/2) of the activity at each region of interest set to encompass the entire kidney and the dilated pelvis was used as an index of quantitative analysis of UPJ obstruction. Initial T 1/2s of 32 kidneys with suspected UPJ obstruction were significantly longer than those of 37 without obstruction. T 1/2s of kidneys which had undergone pyeloplasty decreased promptly after surgery whereas those of units followed up without surgery decreased more sluggishly. These findings demonstrate that a standardized diuretic renographic analysis with T 1/2 can reliably assess infantile hydronephrosis with UPJ obstruction and be helpful in making a decision on surgical intervention.
OBJECTIVES: This study investigated the cycle length changes preceding the spontaneous onset of torsade de pointes in patients with acquired prolonged ventricular repolarization. BACKGROUND: Torsade de pointes is a polymorphic ventricular tachycardia generally associated with prolonged ventricular repolarization. Because torsade de pointes is not inducible by programmed electrical stimulation, quantitative analysis of Holter recordings of spontaneous episodes may clarify the mechanisms favoring the onset of torsade de pointes in actual clinical conditions. METHODS: The digitized Holter recordings of 12 patients were analyzed by a computerized Holter system (ATREC). All arrhythmias were grouped according to three classes: 1) isolated premature ventricular beats (n = 47,147, mean/patient [+/- SD] 3,929 +/- 11,571); 2) salvos of 2 to 4 consecutive beats (n = 2,003, mean/patient 167 +/- 359); 3) torsade de pointes > or = 5 beats (n = 105, mean/patient 9 +/- 11). For each patient and class of arrhythmias, six variables were computed from the 10 min and the 10 cycles preceding the event onset. RESULTS: A significant heart rate increase in the last minute (p < 0.01) and typical oscillatory short-long-short cycle length sequences preceded the onset of arrhythmias, with greater oscillation preceding torsade de pointes than salvos and premature ventricular beats. The cycle lengths preceding the onset were highly correlated with the class of arrhythmias (r = 0.65, p < 0.005) and allowed the correct classification of 69% of events by discriminant analysis (p < 0.0001). A significant negative correlation was observed between the duration of torsade de pointes and the mean length of the initial cycles (r = -0.62, p < 0.001), indicating that longer torsade de pointes had a faster rate than that at onset. CONCLUSIONS: In patients with acquired prolonged repolarization, the spontaneous onset of ventricular arrhythmias was preceded by an increasing heart rate in the last minute and escalating oscillatory "short-long-short" cycle length patterns, with greater oscillations preceding torsade de pointes than salvos and isolated ventricular beats. These findings suggest that adrenergic- and pause-dependent mechanisms (possibly inducing afterdepolarizations and triggered activity) may have a synergetic role in the genesis of complex ventricular arrhythmias associated with delayed ventricular repolarization.