A method to calibrate fetal µCT scans using histological sections from the same specimens.

Craniofacial morphogenesis is an intricate developmental process in 3D, which therefore merits visualization and investigation in 3D. To better understand the process, we utilize µCT imaging, and describe a method to calibrate each cone beam µCT individually. Calibration is necessary, because during development, fetuses undergo tissue differentiation, which affects the acquisition process for radiographic images. Additionally, tissue fixation and conservation agents may influence the physical properties of the specimens and may affect image acquisition. After taking a µCT scan from each specimen, we separated a horizontal slice from each neck (which is inconsequential to our question with relation to the whole head). These neck specimens were prepared as horizontal histological serial sections and stained. With these as a reference, the µCT visualization parameters could be adjusted until they matched the selected virtual section planes, which correspond exactly to the planes of the histological sections with a precision (pixel size) of 0.69µm.

Improving surgical outcomes through benchmarking.

Benchmarking is a popular quality-improvement tool in economic practice. Its basic principle consists of identifying the best (the benchmark), then comparing with the best, and learning from the best. In healthcare, the concept of benchmarking or establishing benchmarks has been less specific, where comparisons often do not target the best, but the average results. The goal, however, remains improvement in patient outcome. This article outlines the application of benchmarking and proposes a standard approach of benchmark determination in surgery, including the establishment of best achievable real-world postoperative outcomes. Parameters used for this purpose must be reproducible, objective and universal. A systematic approach for determining benchmarks enables self-assessment of surgical outcome and facilitates the detection of areas for improvement. The intention of benchmarking is to stimulate surgeons' genuine endeavour for perfection, rather than to judge centre or surgeon performance.

Inequality in mortality decreased among the young while increasing for older adults, 1990-2010.

Many recent studies point to increasing inequality in mortality in the United States over the past 20 years. These studies often use mortality rates in middle and old age. We used poverty level rankings of groups of U.S. counties as a basis for analyzing inequality in mortality for all age groups in 1990, 2000, and 2010. Consistent with previous studies, we found increasing inequality in mortality at older ages. For children and young adults below age 20, however, we found strong mortality improvements that were most pronounced in poorer counties, implying a strong decrease in mortality inequality. These younger cohorts will form the future adult U.S. population, so this research suggests that inequality in old-age mortality is likely to decline.

[Health education for chronic liver and pancreatic diseases: Discussion groups "chronic liver disease" (CL), "chronic pancreatic disease" (CP)]. / Schulung bei Leber- und Pankreaserkrankungen: Gesprächsgruppe "Chronische Lebererkrankungen" (CL) /"Chronische Pankreaserkrankungen" (CP).

The article deals with our experience in the educational programs for patients with chronic liver diseases and chronic pancreatic diseases.The structure of our educational programs is modular, the main event is the discussion group "Chronic Liver Diseases" (CL) (2 meetings are headed by physicians, 1 by psychologists, duration in all 3,6 hours) respectively the discussion group "Chronic Pancreatic Diseases" (CP) (2 meetings headed by physicians, 1 by nutrition consultants, duration in all 2,6 hours). As needed, additional modules can be added, for example specific seminars in case of alcohol abuse or educational programs for diabetics. The average number of participants is between 7 - 15 persons in the discussion group CL, respectively 11 - 20 in the discussion group CP. An overhead projector is used to address the various topics, the patients are encouraged to actively participate in the discussion. The contents can be found in a structured curriculum and focus on anatomical basics, diagnostic and therapeutic aspects, physical fitness and performance in professional life.

Screening, identification and quantitation of benzodiazepines in serum by solid phase extraction on a cyanopropyl phase using high performance liquid chromatography and photodiode array detection.

A screening procedure for benzodiazepines in serum is presented. The compounds are extracted over a cyanopropyl phase followed by identification and determination via high performance liquid chromatography and photodiode array detection. Diazepam (CAS 439-14-5), flunitrazepam (CAS 1622-62-4), nordazepam (CAS 1088-11-5), oxazepam (CAS 604-75-1) and temazepam (CAS 846-50-4) are quantified. Retention data of 42 benzodiazepines and their metabolites as well as hydrolysis products are listed. This assay leads to an expansion of a known systematic toxicological analysis system to identify pharmaceuticals, drugs of abuse and pesticides.

Aerosolized amiloride: dose effect on nasal bioelectric properties, pharmacokinetics, and effect on sputum expectoration in patients with cystic fibrosis.

Aerosolized amiloride normalizes the excessive sodium absorption cystic fibrosis (CF) respiratory epithelium. The aims of this study were to assess the dose-effect relationship and the duration for which amiloride inhibits Na+ transport, to determine acute and chronic pharmacokinetics, and to test the effect of acute aerosolized amiloride on the amount of sputum expectorated. The effect of inhaled amiloride was assessed principally by nasal potential difference (PD) measurements. Amiloride serum levels were measured in 23 patients after inhalation of different doses of aerosolized amiloride. Twenty CF patients inhaled amiloride (10(-3)M) or a placebo in a double-blinded, randomized order, and sputum production was quantitated. The results of this study showed that maximal initial PD inhibition was achieved by 6 x 10(-3)M of amiloride. The duration of inhibition of PD (effective time until return to 50% delta PD [ET50] after nasal administration) was dose dependent (10(-3)M, 39 +/- 0.8 minutes; 10(-2)M; 133 +/- 14 minutes). Amiloride serum levels were below 2.5 ng/ml in 20 of 28 patients; levels were above 5 ng/ml only within 4 hours after high dose inhalation (10(-2)M). In the double-blinded, crossover study, more sputum was expectorated after amiloride inhalation as compared with that after a placebo (P < 0.05). In conclusion, the bioelectric effects of amiloride and serum levels after inhalation are dose dependent, and amiloride is effective at inducing sputum expectoration in CF.

[The determination of a gross utilization of 15N-lysine in laboratory rats. 2. Comparative testing with an antibiotic supplement to the diet]. / Untersuchungen zur Bestimmung einer Bruttoverwertung von 15N- Lysin an Versuchsratten. 2. Mitteilung. Vergleichende Prüfung bei einer Antibiotikazulage zur Diät.

Wistar rats of a live weight of about 100 g received in 26 groups (4 animals/group) diets, each with a different lysine content. The rations given supplied the animals with 75%, 100% or 125% lysine of the calculated requirement. The source of protein in the diets was: barley (B), wheat (W), wheat gluten (WG), isolated soybean protein (assay protein) (S) or soybean meal (SM). For WG and S only the lysine levels 100% and 125% (SM = 116% and 125%) could be achieved. All diet groups were fed for 10 days with and without antibiotics (7 g Nebacitin/kg feed-DM). During the 7-day-period of the main experiment all 24 rations were supplemented with 0.5 g 15N-lysine/kg DM (48.3 atom-% 15N-excess, alpha-aminogroup 95% 15N-labelled). The nitrogen balance was improved only after feeding antibiotics with the diet S 100. It may be supposed that Nebacitin saved the second limiting amino acid methionine against microbial degradation in the digestion tract. The biological value (BV) of feed-proteins declined in the case of the diets B and W in the presence of antibiotics because the absorbed nitrogen was higher, this calculation basis for BV was therefore also higher without an improvement of the N-utilization. The 15N-excretion in faeces was significantly lower after feeding the diets B, W and WG with antibiotics. The 15N-excretion in urine was elevated in the most cases of the antibiotic supplement. The determination of a gross utilization of lysine and 15N-lysine resp. in relation to the lysine retention (availability) was not possible, neither using a labelling of diets with 15N-lysine.

[The determination of a gross utilization of 15N-lysine in laboratory rats. 1. Experiment with normal intestinal flora (without antibiotic supplement)]. / Untersuchungen zur Bestimmung einer Bruttoverwertung von 15N-Lysin an Versuchsratten. 1. Mitteilung. Versuch bei normaler Darmflora (ohne Antibiotikazusatz).

Wistar rats of a live weight of about 100 g were divided into 14 groups (5 animals/group). The rations given supplied the animals with 75%, 100% and 125% lysine, which brought about a moderate growth of the animals of approximately 2 g/animal and day achieved by limited feeding. The 3 lysine levels mentioned could be achieved by lysine supplements (L-lysine-HCl) for the following rations: barley (B), wheat (W), and wheat gluten (WG). For isolated soybean protein (assay protein) (S) the lysine levels 100% and 125% and for soybean meal (SM) the levels 116% and 125% could only be achieved. A control group with whole egg ration (W) (with its natural lysine content of 125% of the requirement) were also tested as comparison. During the 10-day period of the main experiment all 14 rations were supplemented with 0.5 g 15N-lysine (alpha amino group, 95% labelled with 15N). The N balance could only be significantly improved by lysine supplements in the rations B, W and SM with the lysine level of 125%. The biologic value of the protein sources was in rations B and WG also significantly improved by the highest lysine supplement. 15N excess (15N') from the deaminated 15N lysine was excreted with diet B rich in crude fibre mainly in faeces (more than 15% of the intake) and only about 10% in urine. With the diets without native crude fibre the excretion quota changed in favour of urine. The following 15N' amounts in per cent of 15N' intake from lysine were excreted in urine and faeces: B 75 = 31.3, B 100 = 30.9, B 125 = 28.0, W 75 = 24.3, W 100 = 32.2, W 125 = 32.6, GW 75 = 18.3, WG 100 = 24.2, WG 125 = 28.1, S 100 = 39.4, S 125 = 50.4, SM 116 = 34.9, SM 125 = 32.9, W 125 = 19.1. 15N excretion in urine and faeces increased in comparable relations in 6 cases of lysine increase levels only. Gross utilization of lysine can only conditionally be quantified by 15N labelled lysine supplement.

Influence of prolonged physical exercise on the erythropoietin concentration in blood.

Erythropoietin (EPO) and red blood cells were studied in 15 well-trained men before and several times after a marathon run. Changes in red blood cells reflected changes of plasma volume. Immediately after the run, red blood cells were increased due to haemoconcentration, whereas 31 h later the values were decreased due to haemodilution. The EPO concentration was increased 3 h, and more impressive 31 h, after the run. This long-lasting increase in EPO concentration after the marathon run would seem to be responsible for the increased red blood cell mass in long distance runners.

[Elimination of secretions in CF patients under amiloride inhalation]. / Sekretelimination bei CF-Patienten unter Amiloridinhalation.

Inhaled amiloride reduces active absorption of sodium of respiratory epithelium in CF patients and so, transiently, diminishes loss of water. 10 CF patients, 8 to 28 years of age, were examined on two days. First day, they inhaled in a randomised order isotonic saline and a solution of amiloride hydrochloride (0.3 mg/ml) one after another, each inhalation taking twenty minutes. Second day, inhalations were performed in an inverse order. To intensify the effect of inhalation, the inhalation procedure was combined with "autogenic drainage", a special kind of physiotherapy. Main criterion for evaluation was the amount of expectorated sputum. Mean increase of sputum during amiloride inhalation in comparison to saline was +50.4%. Patients and physiotherapist observed a liquefaction of secretion and a decrease of coughing by amiloride and a support of physiotherapy. These results suggest a beneficial clinical effect of regular amiloride inhalation in CF patients.

[Determination of a prececal N-absorption from natural feed by 15N-labeled laboratory rats using the isotope dilution method]. / Ermittlung einer präzäkalen N-Resorption aus natürlichen Futtermitteln an 15N-markierten Versuchsratten nach der Isotopenverdünnungsmethode.

60 Wistar rats (5 animals/group) received 12 different feedstuffs over a period of 7 days and were simultaneously labelled with 15N (orally by means of 15NH4Cl in the feed). In the subsequent 5 days faeces were collected in order to determine the apparent and true digestibility of crude protein. On the 13th experimental day the animals were killed 3 hours after the intake of half the daily ration and the atom-% 15N excess (15N') was determined in the TCA-soluble and TCA-precipitable fractions of the blood plasma and the digesta of the 2nd and 3rd thirds of the small intestine. Precaecal N-absorption was calculated with the help of the quotient [formula: see text] the blood plasma and the digesta The following values (in %) were registered in comparison to true N-digestibility (in the following in brackets): casein = 95.8 (99.2), whole egg = 92.1 (97.5), fish meal = 85.8 (93.4), dried skimmed milk = 98.4 (96.1), soybean meal = 79.6 (90.6), assay protein = 94.2 (98.9), wheat = 92.9 (90.7), barley = 84.3 (84.8), yeast, grown on molasses = 85.6 (87.2), yeast, grown on whey = 86.1 (88.5), biomass of liquid manure = 43.6 (68.9), activated sludge = 54.4 (64.1). One can conclude that the isotope dilution technique demonstrated here as an evaluation method is very well suited for the characterization of the N-digestibility of a feedstuff in the small intestine.

Effects of thiopentone and etomidate on median nerve somatosensory evoked responses.

Somatosensory evoked potentials (SEP) after median nerve stimulation were recorded in 40 patients during infusion of either 15 mg/kg bw thiopentone or 1 mg/kg bw etomidate (n = 10) within 15 min and after 0.3 mg/kg bw etomidate (n = 20). Marked alterations of SEP waveforms and changes in latencies were observed in all patients. Central conduction time (CCT) was significantly correlated to plasma thiopentone concentration. Infusion of high doses of thiopentone and etomidate was followed by a complete loss of middle and long latency components. Amplitude of the primary cortical SEP N20 was found to be unchanged after thiopentone and to be increased after etomidate, indicating the synchronizing properties of this drug. A pronounced increase in SEP latencies and CCT and waveform alterations have to be considered during hypnotic drug administration in intensive care medicine and intraoperatively.

Influence of increased catecholamine levels in blood plasma during cold-adaptation and intramuscular infusion on thresholds of thermoregulatory reactions in guinea-pigs.

Catecholamines and some of their metabolites were determined in urine and blood plasma of guinea-pigs before, during and after acclimation to a cold or warm environment. During adaptation to 5 degrees C the amounts of noradrenaline in plasma and 24-h urine samples continuously increased up to 600% compared with values obtained at an ambient temperature of 22 degrees C. Higher levels of dihydroxyphenylglycol and 3-methoxy-4-hydroxyphenylglycol further indicated an increased turnover of noradrenaline during cold adaptation. Acclimation to an ambient temperature of 28 degrees C reduced the peripheral release of noradrenaline in comparison to the release observed at 22 degrees C. Cold-induced increases in metabolic rate and electrical muscle activity both occur at a considerably lower mean body temperature in cold-than in warm-adapted guinea-pigs. The shift of thermoregulatory cold defence reactions to a lower mean body temperature could also be observed in warm-adapted animals after intramuscular infusion of noradrenaline in amounts comparable to those released during cold adaptation. It is concluded that high peripheral sympathetic activity directly or indirectly inhibits noradrenergic neurons in the lower brain stem that modulate the thermoregulatory control system by means of their afferents to the hypothalamus. As a consequence of this peripheral influence the thermoregulatory set point is shifted to a lower mean body temperature.

[Somatosensory evoked potentials under thiopental and etomidate]. / Somatosensorisch evozierte Potentiale unter Thiopental und Etomidat.

The somatosensory evoked potential in response to median nerve stimulation was recorded in 42 patients during infusion of either 15 mg/kgbw thiopentone (TH) or 1 mg/kgbw etomidate (E) within 15 min and before and after injection of 0.3 mg/kgbw etomidate bolus. Cortical and cervical responses were analysed simultaneously and central conduction time (CCT) was calculated. Marked alterations of waveforms and an increase in latency of the primary cortical SEP and of CCT were observed in all patients. Infusion of TH or E was followed by a diminution of middle and long latency components. Amplitude of the cortical N20 was found to be unchanged during and after TH and to be increased after infusion or injection of E, indicating the synchronizing properties of this drug. The cervical SEP (N14) remained entirely unchanged in response to both agents. During hypnotic drug administration a pronounced increase in latencies and CCT as well as a decrease in the number of identifiable peaks has to be considered when SEP monitoring is performed intraoperatively or in intensive care treatment.

[Monitoring long-term therapy with theophylline preparations in children with asthma]. / Die Uberwachung der Langzeittherapie mit Theophyllin-Präparaten bei Kindern mit Asthma.

In 78 children (4 to 17 years of age) with moderate or severe asthma who were additionally treated with sustained-release theophylline preparations, different ways of drug monitoring were examined. Analysis of plasma and saliva theophylline was performed by means of high performance liquid chromatography. Saliva theophylline turned out to permit a reliable prediction of plasma theophylline, if an individual regression is calculated for each patient, basing on 3 simultaneously performed measurements of theophylline levels in saliva and plasma within the therapeutic range of 8 to 20 mg/l. In 25 patients theophylline levels were determined in venous and capillary blood. There was an excellent agreement (r = 0.97). Thus, a convenient monitoring of theophylline treatment in children is possible.

Determination of uracil and thymine and their nucleosides and nucleotides in picomole amounts by gas chromatography mass spectrometry selected ion monitoring.

A stable isotope dilution method is presented by which uracil (Ura) and thymine (Thy) can be determined with high precision and sensitivity at the picomole level utilizing stable isotope dilution and gas chromatography electron impact mass spectrometry in the selected ion monitoring mode. [15N2]Ura and [2H3]Thy served as internal standards. The molecular ions as well as the [M-CH3]+ ion fragments of silylated Ura and Thy (Ura-TMS and Thy-TMS) were suitable for the assay which provides evidence of specificity, if identical results are obtained at both ions. Nucleosides and nucleotides of Ura and Thy were determined following quantitative hydrolysis in 6 N HCl at 180 degrees C for two hours. Other hydrolysis procedures did not give satisfactory results. Levels of free Ura and Thy were measured in human and rat plasma after solvent extraction with a sensitivity of 20-40 pm ml-1 demonstrating ready applicability of the assay method to biological samples. The potential physiological role of circulating Ura and Thy is discussed.

Effects of enzyme induction and inhibition on microsomal oxidation of 1,4-benzodiazepines.

Metabolic oxidative profiles of diazepam (I) were obtained by aromatic C-4'-hydroxylation, N-1-demethylation, and 3-hydroxylation using a supernatant of rat liver. Incubation of 3-methyldiazepam (VI), which suppressed 3-hydroxylation, and N-1-nor-3-methyldiazepam (VII), were used to separately investigate these three oxidative pathways. Treatment of animals with phenobarbital enhanced N-1-demethylation and 3-hydroxylation, and to a variable extent C-4'-hydroxylation. Application of metyrapone reduced metabolite formation by 3-hydroxylation and N-1-demethylation, but had no effect on C-4'-hydroxylation. Metyrapone inhibition was more pronounced following than prior to phenobarbital treatment. C-2-hydroxylation was studied using medazepam (XX) incubations. This pathway was increased by phenobarbital pretreatment and reduced by metyrapone inhibition which was again more pronounced following than prior to phenobarbital pretreatment. These results support earlier conclusions on the heterogeneity of liver microsomes and suggests the presence of different species of hepatic microsomal terminal oxidases. Phenobarbital treatment and metyrapone change the metabolic profile via induction and inhibition, respectively, and, thus, in the case of 1,4-benzodiazepines, the formation of metabolites with varying pharmacological activity. This could become important in clinical situations as a diagnostic mean to determine induction under various treatment or, possibly, during cumulation of metabolites with a long half-life.

Determination of ftorafur and 5-fluorouracil levels in plasma and urine.

A gas chromatographic method was developed for ftorafur (Ft) detection in plasma and urine with a sensitivity of 1 mug/ml. Specific determination of its metabolite 5-fluorouracil (FU) with a sensitivity of 1 ng/ml was achieved by column chromatographic separation from Ft and subsequent gas chromatography-mass spectrometry of bis-silyl-FU in the selected ion mode (GC-MS-SIM) using bis-15N-FU as internal standard. Intravenous injections of 2-14C-Ft and 2',5'-14C-Ft were given to rats and rabbits respectively, and plasma and urine were analyzed for Ft, and 14C activity. Unchanged Ft accounted for most of the 14C activity in plasma, while FU concentrations were below 0.15% and 0.4% relative to Ft concentrations in the rabbit and the rat, respectively. 30-60% of the urinary 14C activity was unchanged Ft and less than 0.2% FU. The significance or low FU levels is discussed in view of the hypothesis that Ft acts as a transport form of its metabolite FU.