Survival outcome of non-small cell lung cancer patients: Comparing results between the database of the Comprehensive Cancer Center Zürich and the Epidemiological Cancer Registry Zurich and Zug.

BACKGROUND: Cancer cases among the population of the canton Zurich, are registered in the Cancer Registry of the cantons of Zurich and Zug (KKR). The Thoracic Oncology Center, founded in 2011 is one of 17 multidisciplinary centers within the Comprehensive Cancer Center Zurich (CCCZ). METHODS: The aim of the current study is to quantify the mortality risk of patients with NSCLC and identify differences on survival and other factors between patients receiving their primary treatment at the CCCZ and those treated elsewhere and registered by KKR. The differential effect between CCCZ and KKR cohorts on survival: overall, by stage, sex and age, is explored. Stratified log-rank and Wilcoxon tests, Cox models and restricted mean survival times (RMST) are estimated. Propensity score matching (PSM) is also used to adjust for confounding factors. RESULTS: Analysis included 848 NSCLC cases from the CCCZ and 1759 from the KKR, diagnosed between January 2011 and December 2015. At a median follow-up of 57 months, overall survival (OS) was significantly superior for patients treated at the CCCZ compared to KKR [Median OS: 36.0 months (95%CI: 31.0-45.0) and 12.0 months (95%CI: 11.0-13.0), respectively, stratified log-rank p < 0.001; adjusted HR = 1.31, (95% CI: 1.18-1.46), difference in RMST up to 72 months: 13.8 months (95%CI: 11.5-16.2), p < 0.001]. The effect of cohort was significant for stages III and IV (overall and also by sex and age). After PSM OS remained significantly superior for patients treated at the CCCZ compared to KKR. CONCLUSIONS: The survival probability for patients in the CCCZ cohort was superior to that of patients in the canton Zürich treated outside the center. This analysis provides further evidence of the importance of the volume of experience and the availability of a multidisciplinary organization and research environment, as delivered by a comprehensive cancer center, on the outcome of patients with NSCLC.

Environmental signatures and effects of an oil and gas wastewater spill in the Williston Basin, North Dakota.

Wastewaters from oil and gas development pose largely unknown risks to environmental resources. In January 2015, 11.4ML (million liters) of wastewater (300g/L TDS) from oil production in the Williston Basin was reported to have leaked from a pipeline, spilling into Blacktail Creek, North Dakota. Geochemical and biological samples were collected in February and June 2015 to identify geochemical signatures of spilled wastewaters as well as biological responses along a 44-km river reach. February water samples had elevated chloride (1030mg/L) and bromide (7.8mg/L) downstream from the spill, compared to upstream levels (11mg/L and <0.4mg/L, respectively). Lithium (0.25mg/L), boron (1.75mg/L) and strontium (7.1mg/L) were present downstream at 5-10 times upstream concentrations. Light hydrocarbon measurements indicated a persistent thermogenic source of methane in the stream. Semi-volatile hydrocarbons indicative of oil were not detected in filtered samples but low levels, including tetramethylbenzenes and di-methylnaphthalenes, were detected in unfiltered water samples downstream from the spill. Labile sediment-bound barium and strontium concentrations (June 2015) were higher downstream from the Spill Site. Radium activities in sediment downstream from the Spill Site were up to 15 times the upstream activities and, combined with Sr isotope ratios, suggest contributions from the pipeline fluid and support the conclusion that elevated concentrations in Blacktail Creek water are from the leaking pipeline. Results from June 2015 demonstrate the persistence of wastewater effects in Blacktail Creek several months after remediation efforts started. Aquatic health effects were observed in June 2015; fish bioassays showed only 2.5% survival at 7.1km downstream from the spill compared to 89% at the upstream reference site. Additional potential biological impacts were indicated by estrogenic inhibition in downstream waters. Our findings demonstrate that environmental signatures from wastewater spills are persistent and create the potential for long-term environmental health effects.

Exposure to moxifloxacin and cytomegalovirus replication is associated with skin squamous cell carcinoma development in lung transplant recipients.

BACKGROUND: Lung transplant recipients (LTR) are at increased risk for squamous cell carcinoma of the skin (SCC), but risk factors (RF) are incompletely understood. OBJECTIVE: To assess associations between exposure to certain medications and viral infections, and subsequent SCC development. METHODS: Retrospective study examining incidence and potential RF for SCC in LTR transplanted from 1992 to 2010 followed up at one centre. Cumulative incidence and Cox proportional hazards regression models were used to evaluate RF in the first year post-transplant for SCC formation during the follow-up. RESULTS: In 205 analysed LTR, 46 patients were diagnosed with SCC during a median follow-up of 4.9 years. The cumulative incidences of first SCC were 16.7% and 34.1%, for 5 and 10 years post-transplantation respectively. Multivariable analysis identified CMV replication (HR 7.69, 95% CI 2.93-20.2, P < 0.001) and moxifloxacin exposure (HR 2.35, 95% CI 1.15-4.81, P = 0.020) during the first year post-transplantation as independent RF for SCC development during follow-up. CONCLUSION: In our cohort, moxifloxacin use and CMV replication during the first year post-transplantation were associated with increased risk for SCC. These two factors could be indicators of over-immunosuppression. Their role in SCC development requires investigations in larger cohorts and prospective studies.

[Stroke occurence in Germany - on the comparability of insurance data and registry-based data]. / Schlaganfallgeschehen in Deutschland - Zur Vergleichbarkeit von Krankenkassen-, Register- und DRG-Daten.

PURPOSE: This article presents epidemiological data regarding stroke frequency in Germany based on nationwide statutory health insurance data (Deutsche BKK) and aims to analyse them in the context of current research. The comparability of the most important resources of stroke frequency data - stroke registers, DRG data and insurance data - is initially discussed in order to assess the presented data adequately. METHODS: The study cohort comes from a population of about 1 000 000 people insured with BKK and consists of all persons who were treated for a stroke in an acute care hospital in 2007 (n = 4,843). Data were subjected to statistical secondary analysis including uni- and bivariate statistics and t tests. Reference studies for the observation period include data from GEK and AOK health insurances, from quality assurances Hessen and Bayern, from the ADSR, and hospital DRG data. The different study types are compared regarding their inclusion/exclusion criteria and the resulting effects on reported prevalences. RESULTS: Different inclusion criteria and accordingly different operationalisations of "stroke" impede the comparability of existing German data resources regarding stroke. The inclusion of TIA, non-traumatic subdural haemorrhage (I62), and the frequency of unspecified strokes (I64) is especially inconsistent. In addition, recurrent strokes and the definition of first-ever strokes are treated differently. The study cohort reveals no major discrepancies regarding aetiological subgroups compared to previous results, only the percentage of women (60.3 %) seems exceptionally high. CONCLUSIONS: The gender effect is attributed to the BKK member structure, and especially the high proportion of women in the older age groups. Discussion of stroke frequency in Germany needs to take structural differences between study types into account. There are two vulnerable groups that tend to be underrepresented: TIA patients with a high risk of recurrent strokes, and high-risk patients who have already had a stroke and are care-dependent, which are often unspecifically coded. In the future, study designs should include the whole range of stroke coding, thus enabling differentiated analyses.

[A cross-sectorial analysis of physio and occupational therapy pathways after stroke]. / Physio- und ergotherapeutische Versorgungsverläufe bei Schlaganfallbetroffenen - von der Akut- über die Rehabilitationsbehandlung bis zur ambulanten Heilmittelversorgung.

PURPOSE: This article examines the provision of physiotherapy and occupational therapy for stroke patients from a cross-sectorial perspective, from acute to rehabilitative care to outpatient services. METHODS: The sample comprises all clients of the Deutsche BKK, a large German health insurance company, who received acute care for stroke in 2007, who survived the initial hospital stay, and who had a secondary diagnosis of motor deficits (n = 1 929). RESULTS: For 60.4% of these stroke patients, no further treatment was provided after acute care. The odds of receiving early rehabilitation treatment while in hospital stay decreased by 1% with each year of life. Only 18.8% of patients received a form of treatment that was largely in line with current recommendations for stroke care, beginning with early rehabilitation and including further treatment in the context of rehabilitation measures or outpatient care. Patients who were in long-term nursing care before stroke were at increased risk of not being placed on this treatment pathway, which has been positively evaluated. 20.7% of patients did not receive any early rehabilitation treatment, but received further rehabilitation treatment and/or outpatient services after hospital discharge. CONCLUSIONS: We recommend that receipt of long-term nursing care should routinely be regarded as a risk factor for underprovision of treatment after stroke (yellow flag).

Influence of altitude on vitamin D and bone metabolism of lactating sheep and goats.

This study investigated the influence of alpine grazing on vitamin D (vitD) and bone metabolism in sheep and goats. Two groups of five adult lactating East Friesian milk sheep and Saanen dairy goats were kept on pastures at 2,000 to 2,600 m a.s.l. (SA: sheep alpine; GA: goats alpine) and 400 m a.s.l. (SL: sheep lowland; GL: goats lowland). The animals were milked twice daily and the milk yield was measured. Blood, milk, skin, and forage samples were collected and the left metatarsi were measured with peripheral quantitative computed tomography. The relative humidity and air temperature were recorded and the ultraviolet B (UVB) radiation was measured with a solar meter at both research stations. In addition, animals from the alpine group were equipped with a global positioning system receiver. The UVB radiation was higher at the alpine station (P<0.05) compared to the lowland station. In contrast, both the relative humidity and the air temperature were higher at the lowland station (P<0.04). The group GA produced more milk than GL (P<0.043). No differences in milk production between SA and SL were detected. Only minor differences between the alpine and lowland species groups were found in the total 25-hydroxyvitamin D (25(OH)D) and 1,25-dihydroxyvitamin D serum concentration and in the 25(OH)D milk concentration. 25-hydroxyvitamin D2 concentration in serum was higher in sheep compared to goats and the 25(OH)D3 concentration in serum increased in all four groups but was higher in the alpine groups during the experiment. In addition, no differences in 7-dehydrocholesterol (7-DHC) concentrations in the skin at high altitude and lowland groups were detectable. However the 7-DHC concentrations in the skin of sheep were less than a tenth of the concentrations in the skin of goats and were nearly not detectable. In both groups SA and SL bone strength index increased during the trial (P=0.043). Bone strength index was lower in GA compared to GL at wk 12 (P=0.047). Mean serum Ca concentrations were higher and P concentrations were lower in the alpine groups than in the lowland groups (P=0.047). In both groups SA and GA the distance travelled increased during the trial. In conclusion, no effect of altitude on vitD status, vitD milk concentration and bone strength could be detected. Both sheep and goats are able to produce vitD in the skin, but sheep depend more on vitD intake with feedstuff, whereas goats rely more on cutaneous vitD production.

Multi-targeted screening of botanicals in food supplements by liquid chromatography with tandem mass spectrometry.

Safety, quality and composition assessments of food supplements based on botanical ingredients are of major concern, as they have usually not been through a rigorous testing process as required for the approval of therapeutic phytopreparations. Therefore, an efficient multi-targeted method was developed to screen selected botanicals of interest in herbal food supplements. Liquid chromatography coupled with a hybrid triple quadrupole linear ion trap was used for this purpose. Botanicals were characterised by means of appropriate biomarkers, which were unambiguously identified by mass spectrometry using an information dependent acquisition experiment which combined a multiple reaction monitoring survey with dependent enhanced product ion scans. During this procedure, product ion scans of targeted analytes were generated at three collision energies and compared with an in-house library of MS/MS spectra acquired from reference standards of all biomarkers. This generic method enables detection, identification and quantification of 98 biomarkers intended to characterise 79 selected plants.

Quantification of near-field heating during volumetric MR-HIFU ablation.

PURPOSE: High-intensity focused ultrasound guided by magnetic resonance imaging has been extensively evaluated during the past decade as a clinical alternative for thermal ablation of tumor tissue. However, the maximal ablation volume is limited by the extensive treatment duration resulting from the small size of the focal point as compared to the average tumor size. Volumetric sonication has been shown to efficiently enlarge the ablated volume per sonication, but remains limited by the temperature increase induced in the skin and fat layers. In this study, multiplane MR thermometry is proposed for monitoring the near-field temperature rise in order to prevent related unintended thermal damage. METHODS: The method was evaluated by performing sonications in the thigh muscle of 11 pigs maintained under general anesthesia. Volumetric ablations were performed by steering the focal point along trajectories consisting of multiple outward-moving concentric circles. Near-field heating was characterized with MR temperature maps and thermal dose maps. The results from the MR measurements were compared to simulations. RESULTS: In this study, the measured maximum temperature rise was found to correlate linearly with the surface energy density within the near field of the beam path with a slope of 4.2 K mm2/J. This simple linear model appears to be almost independent of the trajectory pattern and the sonication depth. The safety limit to avoid lethal damage of the subcutaneous tissues of the porcine thigh was identified to be an absolute temperature of 50 degrees C, corresponding to a surface energy density of 2.5 J/mm2 at 1.2 MHz. CONCLUSIONS: A linear relationship can be established to estimate the temperature increase based on the chosen power prior to ablation, thereby providing an a priori safety check for possible excessive near-field heating using a known surface energy density threshold. This method would also give the clinician the possibility to abort the sonication should excessive near-field temperature rise be seen before fat layer damage or skin burns are inflicted.

[Intraarterial thrombolysis in acute limb ischaemia: alternative treatment and basic principles of further interventions]. / Lokale Lyse bei akuter peripherer Extremitätenischämie: Therapiealternative und Grundlage weiterer Interventionen.

INTRODUCTION: Intraarterial thrombolysis of acute arterial occlusions in the lower limbs is an established therapeutic procedure. However, its value as a primary treatment is discussed controversially and it is mostly seen as a competing procedure to primary vascular surgical interventions. METHODS: From January 2001 to December 2004, we performed a total of 132 intraarterial thrombolysis procedures in 112 patients with acute or subacute ischaemia of the lower limbs. In the majority of the cases, there was an ischaemia of stage I or II a according to Rutherford, a few further cases were in stage II b at the start of treatment. The patients' ages at the time of thrombolysis was 64.7+/-11.3 years (mean value+/-standard deviation, range: 27-91 years). There were 46 female and 86 male patients. RESULTS: The average duration of thrombolysis was 35.2+/-20.8 hours (mean+/-standard deviation, range: 6-142 hours). As median 2 (range: 0-8) angiographic controls were performed. In 100 cases (75.8%), an operation could be avoided by thrombolysis alone or in combination with an supplementary intervention. On the other hand, in 32 cases (24.2%) a subsequent operative therapy including 10 major amputations (7.6%) was necessary. Because of hemorrhagic complications, 15 thrombolysis procedures (11.4%) had to be terminated prematurely but only 2 patients required an inguinal revision. Therapy-related mortality was 0.8% (1 patient). CONCLUSION: In the majority of our patients, primary intraarterial thrombolysis was successful and often led to the discovery of the underlying vascular lesion. It can serve both as a therapeutic alternative to a primary vascular surgical intervention and as the foundation further interventional or vascular surgical therapies although this cannot always be predicted for individual cases.

Involvement of the retinohypothalamic tract in the photic-like effects of the serotonin agonist quipazine in the rat.

Light is the major synchronizer of the mammalian circadian pacemaker located in the suprachiasmatic nucleus. Photic information is perceived by the retina and conveyed to the suprachiasmatic nucleus either directly by the retinohypothalamic tract or indirectly by the intergeniculate leaflet and the geniculohypothalamic tract. In addition, serotonin has been shown to affect the suprachiasmatic nucleus by both direct and indirect serotonin projections from the raphe nuclei. Indeed, systemic as well as local administrations of the serotonin agonist quipazine in the region of the suprachiasmatic nucleus mimic the effects of light on the circadian system of rats, i.e. they induce phase-advances of the locomotor activity rhythm as well as c-FOS expression in the suprachiasmatic nucleus during late subjective night. The aim of this study was to localize the site(s) of action mediating those effects. Phase shifts of the locomotor activity rhythm as well as c-FOS expression in the suprachiasmatic nucleus after s.c. injection of quipazine (10 mg/kg) were assessed in Lewis rats, which had received either radio-frequency lesions of the intergeniculate leaflet or infusions of the serotonin neurotoxin 5,7-dihydroxytryptamine into the suprachiasmatic nucleus (25 microg) or bilateral enucleation. Lesions of intergeniculate leaflet and serotonin afferents to the suprachiasmatic nucleus did not reduce the photic-like effects of quipazine, whereas bilateral enucleation and the subsequent degeneration of the retinohypothalamic tract abolished both the phase-shifting and the FOS-inducing effects of quipazine. The results indicate that photic-like effects of quipazine are mediated via the retinohypothalamic tract.

Binding of correolide to the K(v)1.3 potassium channel: characterization of the binding domain by site-directed mutagenesis.

Correolide is a novel immunosuppressant that inhibits the voltage-gated potassium channel K(v)1.3 [Felix et al. (1999) Biochemistry 38, 4922-4930]. [(3)H]Dihydrocorreolide (diTC) binds with high affinity to membranes expressing homotetrameric K(v)1.3 channels, and high affinity diTC binding can be conferred to the diTC-insensitive channel, K(v)3.2, after substitution of three nonconserved residues in S(5) and S(6) with the corresponding amino acids present in K(v)1.3 [Hanner et al. (1999) J. Biol. Chem. 274, 25237-25244]. Site-directed mutagenesis along S(5) and S(6) of K(v)1.3 was employed to identify those residues that contribute to high affinity binding of diTC. Binding of monoiodotyrosine-HgTX(1)A19Y/Y37F ([(125)I]HgTX(1)A19Y/Y37F) in the external vestibule of the channel was used to characterize each mutant for both tetrameric channel formation and levels of channel expression. Substitutions at Leu(346) and Leu(353) in S(5), and Ala(413), Val(417), Ala(421), Pro(423), and Val(424) in S(6), cause the most dramatic effect on diTC binding to K(v)1.3. Some of the critical residues in S(6) appear to be present in a region of the protein that alters its conformation during channel gating. Molecular modeling of the S(5)-S(6) region of K(v)1.3 using the X-ray coordinates of the KcsA channel, and other experimental constraints, yield a template that can be used to dock diTC in the channel. DiTC appears to bind in the water-filled cavity below the selectivity filter to a hydrophobic pocket contributed by the side chains of specific residues. High affinity binding is predicted to be determined by the complementary shape between the bowl-shape of the cavity and the shape of the ligand. The conformational change that occurs in this region of the protein during channel gating may explain the state-dependent interaction of diTC with K(v)1.3.

Investigation of the soil-plant transfer of primordial radionuclides in tomatoes by low-level gamma-ray spectrometry.

The paper presents actual data from investigations of the soil-plant transfer of the primordial radionuclides 40K, 238U, 226Ra, 210Pb and 227Ac for tomatoes growing at soils from former uranium mining areas. The analysis were carried out using low-level gamma-ray spectrometry in a 47 m deep underground laboratory. For tomato fruits transfer factors of (0.0007 +/- 0.0006) for 235U, (0.0021 +/- 0.0017) for 226Ra, (0.0015 +/- 0.0009) for 210Pb and (0.0018 +/- 0.0012) for 227Ac were obtained. The investigation of the soil-plant transfer by low-level gamma-ray spectrometry is often limited by the Compton-continuum from the always present high-energy gamma-ray emitter 40K.

Severe HDN due to anti-Ce that required exchange tranfusion.

BACKGROUND: Rh system antibodies are commonly encountered in blood bank practice as well as during pregnancy. Nevertheless, no examples of anti-Ce (RH7) have been reported as a cause of HDN that requires exchange transfusion. CASE REPORT: A 38-year-old woman in her fourth pregnancy was typed as blood group O D+, C-, c+, E+, e-. Anti-C and anti-e were detected in her serum during a routine prenatal work-up. Further evaluation, including flow cytometric analysis, revealed the presence of a strong anti-Ce and a weak anti-e. Her partner was typed as group A D+, C+, c-, E-, e+. A seemingly healthy male infant was delivered at 40 weeks of gestation. The infant's RBCs were typed as group O D-, C+, c+, E+, e+ with a positive DAT (titer 128). Twenty-five hours after birth, the baby had to be transferred to the neonatal intensive care unit because of rapidly rising total serum bilirubin. Despite intensive treatment, including double phototherapy, albumin infusion, and the administration of furosemide and IVIG, the total serum bilirubin level increased during the following day and exchange transfusion with 2 units of type O D-, C-, c+, E+, e- had to be performed; this resulted in a prompt decrease in total serum bilirubin without relapse. CONCLUSION: Anti-Ce caused severe HDN requiring exchange transfusion. This highlights the need for a close follow-up throughout pregnancy if unexpected RBC antibodies are present, to permit the provision of compatible blood in case of a rare antibody.

Measurements of activation induced by environmental neutrons using ultra low-level gamma-ray spectrometry.

The flux of environmental neutrons is being studied by activation of metal discs of selected elements. Near the earth's surface the total neutron flux is in the order of 10(-2) cm(-2)s(-1), which gives induced activities of a few mBq in the discs. Initial results from this technique, involving activation at ground level for several materials (W, Au, Ta, In, Re, Sm, Dy and Mn) and ultra low-level gamma-ray spectrometry in an underground laboratory located at 500 m.w.e., are presented. Diffusion of environmental neutrons in water is also measured by activation of gold at different depths.

Developmental stage- and germ cell-regulated expression of a calcium-binding protein mRNA in mouse Sertoli cells.

There is considerable evidence that germ cells, mainly spermatocytes and spermatids, contribute to the regulation of Sertoli cell activity. We developed an in vitro system to investigate the genes involved in Sertoli cell-germ cell interaction in the mouse by using the differential mRNA display technique. One of the isolated differentially expressed genes, named calgizzarin, belongs to the family of S100 calcium-binding proteins and shows a decreased expression in Sertoli cell-germ cell cocultures compared to cultured Sertoli cells alone. Calgizzarin is expressed in all adult tissues examined, including testis and ovary; however, a high mRNA level for calgizzarin in mouse testis is maintained until day 15 of postnatal development and then declines dramatically, whereas the expression pattern in the ovary remains constantly high. Furthermore, Northern blot studies on testicular RNA from different mouse mutants with defects in spermatogenesis revealed that high levels of calgizzarin transcripts can only be detected in testes of mouse mutants with either no germ cells or primary spermatocytes, but only weak signals for calgizzarin are observed in testes of mutants containing spermatids. In addition, using both RT-PCR analysis and whole-mount in situ hybridization on dissected gonads it was demonstrated that mouse calgizzarin expression starts at 13.5 dpc in the prenatal male gonad and at 16.5 dpc in the embryonic ovary, respectively. The mouse calgizzarin gene was localized on mouse chromosome 5, region E-F. Taken together, our results indicate that calgizzarin expression could be repressed by factors originated from pachytene spermatocytes and/or spermatids.

Changes in cytoplasmic ATP concentration parallels changes in ATP-regulated K+-channel activity in insulin-secreting cells.

Changes in cytoplasmic ATP concentration were monitored in intact insulin-producing cells and correlated to changes in the activity of ATP-sensitive K+-channels (KATP channels). Luciferase was introduced into HIT M2.2 cells and whole pancreatic islets by transient expression of firefly (Photinus pyralis) luciferase cDNA. In transfected cells, extracellular addition of luciferin increased the luminescence signal to a maximum within 50-120 s. Addition of 1 microM of the mitochondrial uncoupler FCCP decreased the luminescence, an effect partly reversed upon withdrawal of the compound. High concentrations of glucose increased cytoplasmic free ATP concentration. Changes in the luminescence signal were accompanied by changes in activity of the ATP-sensitive K+-channel. Transfection per se did not deteriorate cell function, as verified by experiments showing similar changes in cytoplasmic free Ca2+-concentration, [Ca2+li, in both transfected and non-transfected cells. By measuring the cytoplasmic ATP concentration and KATP channel activity under similar experimental conditions, it was possible to establish, for the first time, a direct relationship between these two parameters. This indeed suggests that the cytoplasmic ATP concentration has a crucial role in the regulation of KATP channel activity under physiological conditions.

Extraction of artemisinin and artemisinic acid from Artemisia annua L. using supercritical carbon dioxide.

Artemisinin (an antimalaric compound) and its major precursor artemisinic acid, isolated as the active principles of the medicinal plant Artemisia annua L., were extracted by supercritical fluid extraction (SFE) and analyzed by supercritical fluid chromatography (SFC) using a capillary column, coupled with a flame ionization detector (FID). With optimized operating conditions, artemisinin and artemisinic acid were quantitatively extracted at a flow-rate of 2 ml min-1 in less than 20 min. The supercritical fluid was composed of carbon dioxide and 3% methanol with temperature and pressure fixed at 50 degrees C and 15 MPa, respectively. From the kinetic curves, it appears that the extraction of artemisinin is not limited by the diffusion of the analyte from the plant into the extraction fluid but rather by the elution process. These conditions avoided degradation of the analyte and gave clean extracts ready to be analyzed by SFC. The SFE-SFC-FID method was successfully applied to six samples of A. annua containing various concentrations of artemisinin and artemisinic acid. Results were compared with two conventional liquid solvent extraction processes.

Flow cytometric analyses of the subclasses of red cell IgG antibodies.

We report on flow cytometric IgG subclass determinations of red cell antibodies using polyclonal FITC-labeled antibodies. The limit of detection of this method was 1 ng anti-D per 1 x 10(7) red cells. The inter- and intra-assay coefficients of variance were 8.2 and 2.3%, respectively. In 8 newborns with a positive direct antiglobulin test (DAT) in the gel centrifugation test (GCT), due to ABO antibodies, IgG1 was detected in all and IgG2 additionally in 4 of these cases. In 5 severe cases of hemolytic disease of the newborn (HDN) due to anti-D, large amounts of IgG1 were found, and in 3 of these 5, IgG3 in combination with IgG1. In 8 mild or moderate HDN cases (4 anti-D, 2 anti-E, 1 anti-Fya, 1 anti-Jka), phototherapy sufficed, and IgG1 was the only antibody. In 7 adult patients with malignant lymphoma and a positive DAT (GCT), only small amounts of IgG1 red cell autoantibodies could be demonstrated by flow cytometry. In 5 further patients with malignant lymphoma, a positive DAT, and severe hemolytic anemia, large amounts of IgG1 autoantibodies were found and IgG3 was also present in 3 of these cases. Flow-cytometric determination of IgG subclasses may be a useful tool in immunohematology, since subclass determinations were possible in all of these cases. This method is suited for clinical routine and offers the possibility of sufficient standardization.