New sonographic method for fetuses with a large abdominal circumference improves fetal weight estimation.

PURPOSE: Birth weight (BW) is an important prognostic parameter for neonatal morbidity and mortality. Commonly used weight formulas lack accuracy, especially at the lower and upper end of the fetal weight range. Fetal abdominal circumference (AC) as part of most of the commonly used equations has the greatest impact on weight estimation. It has been shown that formulas specifically designed for a small fetal AC can improve weight estimation. The aim was to find out whether a new formula specifically designed for fetuses with a large AC may also improve weight determination. MATERIALS AND METHODS: The study included 830 singleton pregnancies. The inclusion criteria were ultrasound examination with complete biometric parameters and an AC ≥ 36.0 cm within 7 days of delivery, and an absence of structural or chromosomal malformations. Two "best-fit" formulas were derived by forward regression analysis. The accuracy of the new formulas was compared with commonly used weight equations using percentage error (PE), absolute percentage error (APE), limits of agreement (LOA) and cumulative distribution. RESULTS: New formula I had no systematic error while new formula II and the routine methods significantly overestimated fetal weight. The medians of the APE were the lowest among the new equations (5.77 and 7.25). The new formulas also demonstrated the narrowest LOA. Importantly, at all discrepancy levels (5 %, 10 %, 15 %, and 20 %), new formula I included significantly more cases than the commonly used methods. CONCLUSION: These specifically designed equations help to improve fetal weight estimation for fetuses with an AC ≥ 36.0 cm. For optimal weight estimation, we recommend using new formula I.

New sonographic method for fetuses with small abdominal circumference improves fetal weight estimation.

PURPOSE: Accurate estimation of fetal weight is a valuable tool for determining further obstetric management. Commonly used weight formulas lack accuracy, even though some equations appear to be favorable within defined weight ranges. However, due to the fact that fetal weight is not known in advance, it is not always clear which formula is suitable. In most of the commonly used equations, the fetal abdominal circumference (AC) is not only included but also has the greatest impact on weight estimation. The aim of our study was to develop and evaluate a new formula specifically designed for a small fetal AC in order to improve weight estimation. MATERIALS AND METHODS: The study included 323 pregnancies. The inclusion criteria were singleton pregnancy, ultrasound examination with complete biometric parameters and an AC ≤ 29.0 cm within 7 days of delivery, and an absence of structural or chromosomal malformations. Two "best-fit" formulas were derived by forward regression analysis. Finally, the accuracy of the new formulas was compared to commonly used weight equations by using the percentage error, absolute percentage error (APE), limits of agreement (LOA) and cumulative distribution. RESULTS: Contrary to the routine methods, which significantly underestimated fetal weight, the new formulas did not have a systematic error. The medians of the APE were the lowest (7.13 and 7.16) when compared to other equations. Moreover, the new formulas demonstrated the narrowest LOA. At all discrepancy levels (5%, 10%, 15%, and 20%), the new formulas included significantly more cases than the commonly used methods. CONCLUSION: The specifically designed equations help to improve fetal weight estimation for fetuses with an AC ≤ 29.0 cm. For optimal weight estimation, we recommend using the new formula II.

Fe(II)-catalyzed imidation of allyl sulfides and subsequent

Allyl aryl sulfides 1 and 5 were shown to undergo an imidation/[2,3]-sigmatropic rearrangement reaction upon treatment with N-tert-butyloxycarbonyl azide (BocN3) and catalytic amounts of FeCl2 in CH2Cl2. The N-Boc-protected N-allyl sulfenamides 3 and 21 were obtained in yields between 48 and 75% (12 examples). Whereas the reaction is well suited for the transformation of alpha-unbranched sulfides to alpha-branched sulfenamides, the enantiomerically pure alpha-branched sulfides 10 and 13 reacted sluggishly. The corresponding sulfenamides 22 and 23 were obtained in only moderate enantiomeric excess (36-39% ee). A reaction mechanism is proposed that postulates the intermediacy of an N-Boc-substituted Fe(IV)-nitrene complex 14 acting as the imidation reagent in the catalytic cycle. Possible side reactions are discussed. The benzenesulfenamides 3 were further converted into N-Boc-N-allylamines 4 by removal of the phenylsulfanyl group. Bu3SnH in benzene was found to be the reagent of choice for the deprotection of alpha-branched amines that bear a secondary allyl substituent (five examples, 71-93% yield). This method failed for the alpha-branched amines 3i-k with a tertiary allyl substituent. The phenylsulfanyl group was finally removed with P(OEt)3/NEt3 in CH2Cl2 (three examples, 43-62% yield).

Cell-cycle dependent micronucleus formation and mitotic disturbances induced by 5-azacytidine in mammalian cells.

5-Azacytidine was originally developed to treat human myelogenous leukemia. However, interest in this compound has expanded because of reports of its ability to affect cell differentiation and to alter eukaryotic gene expression. In an ongoing attempt to understand the biochemical effects of this compound, we examined the effects of 5-azacytidine on mitosis and on micronucleus formation in mammalian cells. In L5178Y mouse cells, 5-azacytidine induced micronuclei at concentrations at which we and others have already reported its mutagenicity at the tk locus. Using CREST staining and C-banding studies, we showed that the induced micronuclei contained mostly chromosomal fragments although some may have contained whole chromosomes. By incorporating BrdU into the DNA of SHE cells, we determined that micronuclei were induced only when the compound was added while the cells were in S phase. Microscopically visible effects due to 5-azacytidine treatment were not observed until anaphase of the mitosis following treatment or thereafter. 5-Azacytidine did not induce micronuclei via interference with formation of the metaphase chromosome arrangement in mitosis, a common mechanism leading to aneuploidy. Supravital UV microscopy revealed that chromatid bridges were observed in anaphase and, in some cases, were sustained into interphase. In the first mitosis after 5-azacytidine treatment we observed that many cells were unable to perform anaphase separation. All of these observations indicate that 5-azacytidine is predominantly a clastogen through its incorporation into DNA.

[Is the post-therapeutic dosimetry of patients with short-term hospitalization after 131I therapy sufficiently reliable?]. / Ist die posttherapeutische Dosimetrie bei verkürzter stationärer Verweildauer der Patienten nach 131-I-Therapie ausreichend zuverlässig?

AIM: According to the new recommendations of the Federal German Radiation Protection Committee (SSK) for patient discharge, that were published in April 1997, patients can be discharged after radioiodine therapy with a radiation exposure of less than 1 mSv per year in 2 m distance. The aim of this study was to evaluate whether the measurement of the achieved dose was different 48 hours after application of I-131 and after an interval of one week. The study was planned in order to ensure quality management in the follow-up of the patients. METHOD: In a prospective study 115 patients were followed, that were treated for a benign thyroid disease. The patients were discharged 48 hours after the intake of I-131 after falling short of the exposure limit. All patients were measured at discharge and about 8 days later with the same uptake facility. RESULTS: Because of similar results focal and disseminated forms of benign thyroid diseases could be analysed together. The calculated doses 8 days after the discharge were higher than the values at the time of discharge. The doses were underestimated about 4% with an standard error of 15%. CONCLUSION: The error in early dosimetric measurements is small in comparison to the overall uncertainty in therapy and uptake dosimetry. A valid dosimetry and sufficient quality management can be guaranteed even for an early discharge.

[Measurement of incorporation in family members of radioiodine therapy patients after therapy of benign thyroid diseases]. / Inkorporationsmessungen bei Angehörigen von Radioiodtherapiepatienten nach Therapie benigner Erkrankungen der Schilddrüse.

AIM: Patients exhale I-131 after radioiodine therapy. In this study we quantify the amount of radioactivity and resulting thyroid doses found in people living in close contact to patients treated with I-131 after their release from a therapy ward. METHODS: For 31 relatives of 25 patients treated with I-131 the incorporation was monitored using the thyroid probe of a whole body counter. These values are used for a determination of thyroid doses. RESULTS: 11 of the 31 monitored persons had a thyroid activity of less than the minimal detectable activity of 13 Bq. The mean value of the remaining 20 people was 104 Bq in the thyroid resulting in a mean thyroid dose of 0.2 mSv (Maximum: 2 mSv). CONCLUSION: The intake of I-131 for persons in close contact to patients after dismissal from a therapy ward is low. In no case an effective dose exceeding 1 mSv was observed.

Does an individual estimation of halflife improve the results of radioiodine therapy of Graves' disease?

AIM: The impact of our dosimetry concept on radioiodine therapy success in Graves' disease (GD) was analysed. Three questions arised: Did individual estimation of pretherapeutic halflife improve therapeutic success? Did individual dosimetry result in accurate dose calculation? Did antithyroid medication have a measurable influence on therapeutic success under the prevailing conditions? METHODS: 126 consecutive patients were treated with 200 Gy I-131 in our therapy ward for GD and followed-up six to nine months after therapy. Success quote was assessed using a standardized protocol and treatment was classified as successful when the patient was eu- or hypothyroid, or unsuccessful when he or she presented with a suppressed TSH-level or in hyperthyroid condition after antithyroid medication withdrawal. Antithyroid medication, activity I-131, dose, concentration of fT3 and fT4, specific delivered dose and halflife were put into a multiple regression model to assess their influence on therapeutic success. In order to assess possible factors disturbing the therapeutic outcome, relevant parameters were analyzed using Logit transformation. RESULTS: Out of 126 patients 84 were classified as successfully treated and 42 (33.3%) as failures. A significant influence on the outcome only was found for thyroid mass. However, therapeutic success appeared to be more distinctly determined by the specific delivered dose using an estimated halflife of 5.5 days (Odds: 10.0, p < 0.001). Accurate intratherapeutic dosimetry did not play a significant role to enhance therapeutic success. Neither did antithyroid medication during radioiodine therapy exert any significant impact. CONCLUSIONS: Measurement of individual intratherapeutic halflife as opposed to an estimate using a standard halflife did not provide improved results concerning the target dose. Retrospectively, the therapeutic outcome on the basis of a measured halflife as compared to a standard halflife did not significantly improve. In addition, no influence of antithyroid medication on therapy success was found.

[Incidence of familial non-medullary thyroid carcinoma in the patient register of the Clinic and Polyclinic of Nuclear Medicine, University of Würzburg]. / Häufigkeit des familiären nicht-medullären Schilddrüsenkarzinoms im Krankengut der Klinik und Poliklinik für Nuklearmedizin der Universität Würzburg.

AIM: In this study the incidence rate of familial non-medullary thyroid carcinoma was investigated in the first and second grade relatives of patients registered at the Clinic and Polyclinic for Nuclear Medicine, University of Würzburg. PATIENTS AND METHODS: In this study 596 patients with differentiated thyroid carcinoma were enclosed, who were treated between 01.01.81 and 31.12.95. The data concerning a familial occurrence were studied by a retrospective survey-based analysis. These data were compared to a literature analysis for familial non-medullary thyroid carcinoma. RESULTS: 14 patients of the 596 patients treated showed a familial occurrence (2.3%). All these patients suffered from papillary thyroid carcinoma. According to the prognostic factors (tumor state, lymph node involvement, metastatic disease) no differences could be evaluated in the different groups (sporadic versus familial non-medullary thyroid disease). CONCLUSION: A familial occurrence of differentiated thyroid carcinomas is not frequently observed, but should be considered due to further genetic diseases.

[Multivariate analysis of factors influencing the effect of radiosynovectomy]. / Multivariate Analyse der Einflussfaktoren auf die Wirkung der Radiosynoviorthese bei entzündlichen Gelenkerkrankungen.

OBJECTIVE: In this prospective study, the time to remission after Radiosynovectomy (RSV) was analyzed and the influence of age, sex, underlying disease, type of joint, and duration of illness on the success rate of RSV was determined. METHODS: A total number of 57 patients with rheumatoid arthritis (n = 33) and arthrosis (n = 21) with a total number of 130 treated joints (36 knee, 66 small and 28 medium-size joints) were monitored using visual analogue scales (VAS) from one week before RSV up to four to six months after RSV. The patients had to answer 3 times daily for pain intensity of the treated joint. The time until remission was determined according to the Kaplan-Meier survivorship function. The influence of the prognosis parameters on outcome of RSV was determined by multivariate discriminant analysis. RESULTS: After six months, the probability of pain relief of more than 20% amounted to 78% and was significantly dependent on the age of the patient (p = 0.02) and the duration of illness (p = 0.05), however not on sex (p = 0.17), underlying disease (p = 0.23), and type of joint (p = 0.69). CONCLUSION: Irrespective of sex, type of joint and underlying disease, a measurable pain relief can be achieved with RSV in 78% of the patients with synovitis, whereby effectiveness is decreasing with increasing age and progress of illness.

Nature of the band gap of In2O3 revealed by first-principles calculations and x-ray spectroscopy.

Bulk and surface sensitive x-ray spectroscopic techniques are applied in tandem to show that the valence band edge for In2O3 is found significantly closer to the bottom of the conduction band than expected on the basis of the widely quoted bulk band gap of 3.75 eV. First-principles theory shows that the upper valence bands of In2O3 exhibit a small dispersion and the conduction band minimum is positioned at Gamma. However, direct optical transitions give a minimal dipole intensity until 0.8 eV below the valence band maximum. The results set an upper limit on the fundamental band gap of 2.9 eV.

Phenomena at the advancing ice-liquid interface: solutes, particles and biological cells.

Ice formation in aqueous solutions and suspensions involves a number of significant changes and processes in the residual liquid. The resulting effects were described concerning the redistribution of dissolved salts, the behaviour of gaseous solutes and bubble formation, the rejection and entrapment of second-phase particles. This set of conditions is also experienced by biological cells subjected to freezing. The influences of ice formation in that respect and their relevance for cryopreservation were considered as well. A model of transient heat conduction and solute diffusion with a planar ice front, propagating through a system of finite length was found to be in good agreement with measured salt concentration profiles. The spacing of the subsequently developing columnar solidification pattern was of the same order of magnitude as the pertubation wavelengths predicted from the stability criterion. Non-planar solidification of binary salt solutions was described by a pure heat transfer model under the assumption of local thermodynamic equilibrium. The rejection of gaseous solutes and the resulting gas concentration profile ahead of a planar ice front has been estimated by means of a test bubble method, yielding a distribution coefficient of 0.05 for oxygen. The nucleation of gas bubbles has been observed to occur at slightly less than 20-fold supersaturation. The subsequent radial growth of the bubbles obeys a square-root time dependence as expected from a diffusion controlled model until the still expanding bubbles become engulfed by the advancing ice-liquid interface. The maximum bubble radii decrease for increasing ice front velocities. The transition between repulsion and entrapment of spherical latex particles by an advancing planar ice-front has been characterized by a critical value of the velocity of the solidification interface. The critical velocity is inversely proportional to the particle radius as suggested by models assuming an undisturbed ice front. The increase of the critical velocity for increasing thermal gradients shows good agreement with a theoretically predicted square-root type of dependence. Critical velocities have also been measured for yeast and red blood cells. The effect of freezing on biological cells has been analyzed for human lymphocytes and erythrocytes. The reduction of cell volume observed during non-planar freezing agrees reasonably well with shrinkage curves calculated from a water transport model. The probability of intracellular ice formation has been characterized by threshold cooling rates above which the amount of water remaining within the cell is sufficient for crystallization.(ABSTRACT TRUNCATED AT 400 WORDS)

Intracellular ice formation: cryomicroscopical observation and calorimetric measurement.

The formation of ice crystals within biological cells is generally deleterious and results in a severe loss of cellular viability and function. With the aim of circumventing this lethal event, the mechanisms of nucleation and their dependence on governing parameters such as temperature, cooling rate and solute and/or additive concentration, and the correlation with the osmotically induced water transport across the cell membrane were investigated. Quantitative low-temperature light microscopy was used for this purpose as it offers the major advantage of studying the dynamics of the involved processes. To substantiate further the visual observations of the morphological changes associated with intracellular ice formation, supplementary studies by differential scanning calorimetry (DSC) were performed under comparable conditions to measure the quantity of water actually transformed into the crystalline state due to the evolution of latent heat. Human lymphocytes were used as a biological model cell. In particular it could be shown that the twitching type of intracellular ice formation which is evident but difficult to observe under the cryomicroscope can be attributed to a liquid-solid phase change within the cells as determined by DSC. Good agreement was obtained between the results measured by both techniques with respect to the following dependencies of governing parameters: the fraction of cells exhibiting intracellular ice determined as a function of the cooling rate shows a sharp demarcation zone with an increase from 0 to 100% at about the same threshold cooling rate. On the other hand, the temperatures at which intracellular ice forms were found to be only weakly dependent on the cooling rate. With respect to the effect of cryo-additive concentration at a fixed value of the cooling rate, the crystallization temperatures were seen to decrease with concentration. The DSC results may hence be regarded as a validation of the microscopic observations.

Quantitative cryomicroscopic analysis of intracellular freezing of granulocytes without cryoadditive.

Purified human granulocytes were frozen in isotonic saline at different constant cooling rates down to -60 degrees C and subsequently thawed on the thermally defined cryostage of a cryomicroscope. Cells monitored on videotape were examined with respect to cooling rate threshold, type, and temperature of intracellular ice formation during cooling and recrystallization during warming. Two apparently different mechanisms of intracellular ice formation (iif) were distinguished during cooling, i.e., "twitching" (no visible ice front) and "darkening" (diffuse ice front). Both types of iif are related to cooling rate and hence also to dehydration. Cooling rate thresholds and temperatures of intracellular recrystallization were determined. It was found that twitching iif occurs just about 6.3 to 7.4 degrees C above the homogeneous nucleation temperature, suggesting that it might be catalyzed by nucleators present within the cells. Darkening iif, on the other hand, was observed at much higher temperatures, i.e., 23.4 to 28.3 degrees C above the homogeneous nucleation temperature, which could possibly indicate a nucleation induced by extracellular ice crystals (at a cooling rate of 30 degrees K/min, however, darkening iif was observed to occur at a temperature lower than that required for twitching iif). The proposed mechanisms of cryoinjury are related to membrane integrity measurements presented in M. W. Scheiwe, Ch. Körber, and S. Englich, Cryo-Letters, 5, 300-306, 1984.

Basic investigations on the freezing of human lymphocytes.

Human lymphocytes were frozen at constant cooling rates in the range 2.4 to 1000 degrees K/min without cryoadditive on the cold stage of a thermally defined cryomicroscope. The volume loss due to water efflux was quantified optically for the cooling rates 2.4, 12, 48, and 120 degrees K/min. The likelihood of the formation of intracellular ice was determined as function of the cooling rate. Intracellular crystallization temperatures were obtained for ice formation during both cooling and rewarming. A theoretical analysis of the cell volume loss during freezing was compared to the experimental data and used for an indirect determination of the water permeability of the cells. A relative optimum of the cooling rate is predicted theoretically under the assumption of a critical level of intracellular salt concentration near the eutectic temperature. The dependence of survival and cooling rate was determined cryomicroscopically by simultaneously applying the FDA/EB fluorescence viability test. The optimal cooling rate of about 35 degrees K/min was also found for 2-ml samples frozen within the range of cooling rates of interest. The results show that for freezing in physiological saline solution (1) the optimum of the cooling rate is theoretically predictable, (2) cryomicroscopical data are significant for freezing of samples of larger volume, and (3) the lethal type of intracellular crystallization is cooling rate dependent and distinguishable from innocuous types.

Thermally defined cryomicroscopy and thermodynamic analysis in lymphocyte freezing.

A cryomicroscope is described which provides the possibility of quantifying the volume loss of cells during freezing, detection of intracellular ice formation during cooling and warming, as well as the determination of viability as function of (constant) cooling rates. The basic mechanisms occurring in cryopreservation have been studied with this system using the human lymphocyte suspended in pure saline as a biological model system; experimentally observed exosmosis during freezing is compared to predictions from a thermodynamic model. Cell volume loss during freezing has been determined experimentally for cooling rates of 2.4, 12, 48, and 120 degrees K/min. Exosmosis also was calculated corresponding to various assumptions regarding the concentration dependence of the hydraulic permeability of the cells. Further calculations of exosmosis are performed for determining the effects of the initial cell volume. The temperatures and transition cooling rate ranges of intracellular ice formation have been determined. On the basis of exosmosis and a lethal level of intracellular salt concentration, a hypothetical relative optimum of the cooling rate is theoretically predicted and compared to the experiments.

Thermally defined cryomicroscopy and some applications on human leucocytes.

A freezing-stage has been developed for use on a standard light-microscope, which can provide reproducible, precisely linear cooling and warming rates in the range from 0.1 to 10,000 K/min. Biological cells in aqueous solutions can be observed during the freeze-thaw cycle; the volume loss due to osmotic efflux of water and the intracellular crystallization of water are detected by video-monitoring. The temperature field generated in the observed samples is comparable to extended cylindrical probes and allows the transfer of cryomicroscopic data to technically used vial geometries. Lymphocytes and granulocytes were observed during freezing using the system described. They were separated and washed, and then frozen on the cold stage of the cryomicroscope at cooling rates ranging from 2 to 500 K/min. Shrinkage of the cells was observed up to 100 K/min and intracellular ice formation could be detected starting at 10 K/min. The results show that human leucocytes show excessive shrinkage up to 36% of their initial volume; the probability of intracellular ice formation exhibits a sharp increase from 10 to 100 K/min where nearly all cells contain ice.

Encapsulation of human erythrocytes by growing ice crystals.

In this study the interaction of human erythrocytes in suspension with a planar ice-liquid interface is investigated. Due to a repulsive van der Waals force repelling the cells from the crystal and an attractive force resulting from the viscous drag of melt flow around the cell a velocity was determined below which the cell is pushed by the growing crystal and above which it is entrapped by the solid phase. The critical velocity for erythrocytes suspended in 0.85 wt% NaCl-D2O solution can be derived from measuring the time periods of pushing and is calculated to be 1.1 microns/s at a temperature gradient of 15.3 K/mm.

Where should the cooling rate be determined in an extended freezing sample?

Computer simulations have been performed to calculate the transient cooling and solidification process at different locations in a flat plate-shaped freezing container filled with isotonic solution (0.9 wt% NaCl in water). The one-dimensional model accounts for the influences of the external cooling conditions, the container wall, the freezing bag, and the nonplanar solidification of an aqueous solution. The cooling rate was found to increase within the sample from the portions adjacent to the inner surface of the bag toward the center. An important result for cryobiological experiments was the fact that the geometrical center of the sample, a commonly used location for the determination of cooling rate, is not representative for the entire volume of the sample. Even worse, the calculations have shown that the center is often the least representative place. As an alternative, the most suitable location for cooling rate measurements has been determined. With the assumed surface cooling and geometry conditions an optimum location at x/(d/2) approximately 2/3 (x is the space coordinate, originating at the inner surface of the freezing bag; d is the sample thickness) has been calculated, i.e., a distance of one-third away from the center and two-thirds from the inside surface of the sample container. Admitting a 50% range of variation, the cooling rate measured at this point represents at least 80% of the entire sample volume. The survival signature, i.e., the functional dependence of cell survival from cooling rate (determined at a single location), for a fictitious cell kind is also influenced by the location of temperature determination: the "optimum" cooling rate seems to be shifted, and the shape of the signature is changed depending on the location where the cooling rate is determined.