Search

Background Safety concerns caused by gadolinium retention call for the development of high-relaxivity gadolinium-based contrast agents (GBCAs) allowing minimal dosing. Purpose To investigate brain gadolinium retention in healthy rats after exposure to gadopiclenol (Elucirem, Guerbet; macrocyclic GBCA) compared with gadobutrol (Gadovist or Gadavist, Bayer; macrocyclic GBCA) and gadodiamide (Omniscan, GE Healthcare; linear GBCA) over 1 year. Materials and Methods In this study conducted between May 2018 and April 2020, 9-week-old healthy Sprague Dawley rats received five injections of either gadopiclenol, gadobutrol, or gadodiamide (2.4 mmol of gadolinium per kilogram of body weight for each), or saline (control animals) over a period of 5 weeks. Rats were randomly assigned to different groups (six female and six male rats per group). MRI examinations were performed before euthanasia at 1, 3, 5, or 12 months after the last injection. Brains were sampled to determine the total gadolinium content via inductively coupled plasma mass spectrometry (ICP-MS), to characterize gadolinium species with size exclusion chromatography (SEC)-ICP-MS, and to perform elemental mapping with laser ablation (LA)-ICP-MS. Mann-Whitney tests were performed on pairwise comparisons of the same time points. Results For both macrocyclic agents, no T1 signal hyperintensities were observed in the cerebellum, and approximately 80% of gadolinium washout was found between 1 month (gadobutrol, 0.30 nmol/g; gadopiclenol, 0.37 nmol/g) and 12 months (gadobutrol, 0.062 nmol/g; gadopiclenol, 0.078 nmol/g). After 12 months, only low-molecular-weight gadolinium species were detected in the soluble fraction. Gadodiamide led to significantly higher gadolinium concentrations after 1 month in the cerebellum (gadodiamide, 2.65 nmol/g

Item does not contain fulltext Background The use of gadolinium-based contrast agents (GBCAs) is linked to gadolinium retention in the skeleton of healthy individuals. The mechanism of gadolinium incorporation into bone tissue is not fully understood and requires spatially resolved analysis to locate the gadolinium. Purpose To compare the quantitative distribution of gadolinium retained over time in rodent femur following the administration of gadodiamide and gadobutrol at three different time points. Materials and Methods In this animal study conducted between May 2018 and April 2020, 108 9-week-old healthy rats were repeatedly injected with either gadodiamide, gadobutrol, or saline solution and were killed 1, 3, or 12 months after the last injection. The femurs of six female and six male rats per each group and time point were collected. Quantitative elemental imaging of gadolinium in longitudinal thin sections was performed on one sample per sex with use of laser ablation inductively coupled plasma mass spectrometry (ICP-MS). Gadolinium concentration was determined with use of ICP-MS on the samples of all animals (six per group). Mann-Whitney U tests were applied on pairwise comparisons to determine potential sex effect and GBCA effect on gadolinium concentrations. Results The highest gadolinium retention was observed in the gadodiamide group (concentration, 97-200 nmol · g(-1)), exceeding the mean concentration in the gadobutrol group (6.5-17 nmol · g(-1)). However, the gadolinium distribution pattern was similar for both contrast agents, showing prominent gadolinium retention at endosteal surfaces, in the bone marrow, and in small tissue pores. Gadolinium distribution in cortical bone changed over time, initially showing a thin rim of higher concentration close to the periosteum, which appeared to grow wider and move toward the interior of the femur over 1 year. Conclusion For both gadolinium-based contrast agents, gadolinium retention in rat bone was initially located close to the periosteum and bone cavities and changed with bone remodeling processes. The relevance to long-term storage of gadolinium in humans remains to be determined. © RSNA, 2022 Online supplemental material is available for this article. 01 februari 2023

The application of ordinary least squares (OLS) linear regression is widely used in order to approximate linear external calibration data. However, the assumption of homoscedasticity is often not considered as a requirement for correct data approximation, which can result in a poor regression fit that is often more prominent in the lower concentration range. Heteroscedasticity in inductively coupled plasma-mass spectrometry (ICP-MS) data has been discussed in literature as an intrinsic problem and was found to be addressed better by the use of weighted least squares (WLS) regression in multiple studies. In this study, the effects of OLS and WLS linear regression models have been investigated for quantitative imaging experiments by means of laser ablation (LA)-ICP-MS using matrix-matched standards. The calibration data produced by this technique was found to be heteroscedastic in all 60 analyzed datasets, which yielded poor regression fits for OLS linear regression. In comparison to conventional ICP-MS analysis, the resulting negative effects were found to become even more visible in imaging LA-ICP-MS due to an inaccurate estimation of the regression line's intercept. Also, the calculation of average concentrations in selected regions of interest (ROIs) yields incorrect quantification results at the lower end of the calibration range. The application of WLS linear regression resulted in an improved goodness of fit (GOF), although the weighting factor should be selected carefully. Besides the reciprocal of the variance of each calibration standard (1/si2), more empirical weighting factors that have been discussed in the literature were also evaluated regarding the GOF.

The plug-and-play hyphenation of UV-laser ablation (LA) and mass spectrometry is presented, using dielectric barrier discharge ionization (DBDI). The DBDI source employed here is characterized by its unique geometry, being directly mounted onto the inlet capillary of a mass spectrometer. In the literature, this particular kind of DBDI source is also referred to as active capillary plasma ionization. It has been commercialized as soft ionization by chemical reaction in transfer (SICRIT) and will be addressed as DBDI in this study. LA-DBDI-MS was used for the direct, molecule-specific and spatially resolved analysis of various solid samples, such as coffee beans and pain killer tablets without extensive sample preparation. The combination of fast washout UV-laser ablation and the principle of the DBDI source used here allowed for highly efficient soft ionization as well as high spatial resolution down to 10 μm for molecular imaging.

© 2020 The Royal Society of Chemistry. This work introduces a new method for immuno-mass spectrometry imaging via quadrupole-based laser ablation-inductively coupled plasma-mass spectrometry instruments that is matched to the abundance of elements in biological tissues. Manipulation of ion-optics and quadrupole mass filter parameters provided increased transmission of low level high-mass elements, which are typically used as labels for antibodies, at the expense of highly abundant endogenous low-mass elements. Transmission of mid-mass elements such as transition metals was only slightly affected. The implications for mass resolution and background signals are critically discussed and signal to noise ratios and imaging capabilities are compared to those obtained from a standard method. This novel approach resulted in a 6-fold improved signal to noise ratio for lanthanides that are routinely used as elemental labels for antibodies to target protein distributions in biological tissues. This increase in signal intensity, enhanced contrasts, lower limits of detection and the potential to improve spatial resolution contributed to enhanced imaging and trace analyses, as demonstrated by imaging murine brain sections of the hippocampal system and substantia nigra.

With the introduction of monoclonal antibodies in immunohistochemistry and im-munocytology the detection of single disseminated tumor cells became achievable. In breast cancer as well as in gastric, c

Short-term and medium-term effects of liming (CaCO3), fertilization [5Ca(NO)3)2·NH4NO3], and acidification on soil bioactivity were measured in a spruce stand in Southern Germany. The experiment was set up in a randomized block design. Acid precipitation lowered the pH, liming increased the pH, while fertilization caused only small alterations in pH values. Significant differences in soil moisture occurred only in the mineral horizons. The soil ATP content of the humus layers decreased in all plots (control included) up to day 100. On all sampling dates, a pronounced decrease in ATP content followed the acidification. Minor decreases in ATP were observed after fertilization, while liming produced no defined effects. Similar trends, but less pronounced, were observed in the mineral horizons. Only a few significant correlations were found between pH values and ATP or between moisture and ATP within a treatment and sampling date.

Individual disseminated epithelial tumour cells were detected in bone marrow aspirates in 41 of 108 patients (37%) with squamous cell cancer of the head and neck region by an immunocytochemical technique based on monoclonal antibodies raised against the cytokeratin No. 19. In the clinical stage I (T1N0M0) tumour cells were detected only in 26.3% of the patients, whereas in stage IV (T4N0M0, T(all)N2-3M0, T(all)N(all)M1) almost twice as many patients (47.7%) presented with tumour cells in the bone marrow. Apparently, grade of differentiation of the tumour (grading) had no influence on the spread of single tumour cells. An influence of the different localisations of the primary tumour on tumour cell spread or the rate of tumour recurrence cannot as yet be discovered. Cytokeratin No. 19 expressing cells were not detectable in the bone marrow of 18 patients with non-malignant disease. Seventy-three patients were included in a follow-up study with a mean observation time of 25 months (range: 4-52 months). The presence of epithelial cells at the time of primary treatment appears to indicate a significantly higher risk of development of local or distant tumour recurrences (p = 0.01). Of 46 patients initially exhibiting no tumour cells in the bone marrow, only 14 had a clinical recurrence. Whereas 17 of 27 patients who presented with tumour cells in the bone marrow developed either a local tumour recurrence or distant metastases in different organs. Patients presenting with bone marrow tumour cells showed a significantly shorter disease-free survival than those without (p = 0.002).(ABSTRACT TRUNCATED AT 250 WORDS)

Background: In advanced renal cell cancer (RCC) interleukin-2 (IL-2) and interferon-α (IFN-α) have shown modest clinical activity as single agents, substantial toxicity being associated with IL-2. For

The pathophysiological mechanisms involved in the development of a spontaneous systemic capillary leak syndrome (CLS) are unknown. In contrast, CLS is a well-known side effect of high-dose interleukin-2 (IL-2) therapy in solid tumors. We report on a patient who developed CLS with high serum levels of endogenous IL-2 under immunosuppressive therapy for chronic graft-versus-host disease (GvHD) after allogeneic bone marrow transplantation (BMT). Generalized edema persisted for 10 weeks. The condition resolved after antibiotic therapy of a septic shock with beta hemolyzing streptococci group A. Thus, a latent infection may alter cytokine homeostasis and may cause CLS in BMT patients.

Advanced gastrointestinal tumors have a poor prognosis and are largely unresponsive to currently available chemotherapeutic drugs. The development of more effective therapies would be aided by better preclinical models. An in vitro multicellular cancer spheroid model was established using a modified liquid overlay technique and compared to the corresponding monolayers and xenografts in immunodeficient mice. Morphological and functional differentiation and expression of cell adhesion molecules were evaluated by histology, immunohistochemistry and flow cytometry. Cell cycle analysis was investigated by PI intercalation and the therapeutic efficacy of new drugs was tested with MTT and TUNEL assay. 77% of the gastrointestinal cancer cell lines reflected growth characteristics of their parental gastric carcinomas in three-dimensional culture. Thus, cell lines derived from peritoneal and pleural carcinomatosis grew as single cells and cell aggregates. Cell lines originated from solid primary and metastatic tumors formed partly or fully compact multicellular spheroids recapitulating the tumor architecture of the respective original tumor. The differentiated phenotype was lost after subcutaneous implantation of the in vitro spheroids in mice. The degree of morphological differentiation was reflected by the levels of mucin and constitutive E-cadherin expression. Heterogeneous changes of various molecules involved in tumor progression (e.g. EGF-R, HER2/neu, EpCAM, CD44s) were observed in 2D and 3D culture. Cell cycle analysis revealed most of the spheroid cells in G0/G1, which is contrary to the corresponding monolayers. These tumorbiological characteristics induced in 3D culture had a profound influence on testing of new drugs. Efficacy, side effects, penetration of a drug into the microtumor as well as various aspects of the pharmacodynamics were determined in the spheroid model. Recapitulating some complexity of their in vivo counterparts, multicellular cancer spheroids represent a physiologically valid model for studying the biology of gastrointestinal tumors, including the potential to test new therapeutic strategies.

The inhibitory effect of different polyphenolic plant compounds on alpha-amylase activity was investigated in vitro. A kinetic assay was performed using 96-well-plates. Acarbose was used as positive control (IC50: 23.2 microM). Some of the tested compounds, occurring in plants traditionally used in anti-diabetic tea species, showed an inhibition of the enzyme in physiological concentrations, e.g. luteolin, tannic acid, and isochlorogenic acid.

Metastasis is the principal event leading to death in cancer patients. However, expression of diagnostic and therapeutic molecules is determined in primary tumors, although they are often completely resectable. Therefore, the present study immunohistochemically compared the expression profile of various prognostic molecules in primary gastric tumors (n = 35) and their synchronous metastases (regional lymph nodes, n = 32, liver, n = 10, peritoneum, n = 17). Lewis X and ICAM-1 revealed a similar expression pattern in primary and metastatic lesions. Contrary, in comparison to the primary cancers, CD44 expression was upregulated in the metastatic tumors independently of their localisation (p = 0.009). E-cadherin and LFA-3 expression were preferentially increased in distant metastases, but not in locally advanced gastric tumors (liver: E-cadherin, p = 0.001; liver and peritoneum: LFA-3, p = 0.0067). These data suggest the consideration of the metastatic protein profile for the selection of cancer patients in new molecular therapeutic strategies.

In a series of gastric cancer patients twenty-eight percent of the primary tumors expressed high levels of the Lewis carbohydrate. This correlated significantly with clinicopathological parameters of advanced disease (tumor size of50 mm, M1 stage, UICC stage IIIB/IV). In multivariate analysis, high Lewis expression was found to be a new independent factor of poor prognosis. In addition, Lewis was expressed on solid metastases and micrometastatic cells originating from Lewis positive primary tumors. The data suggest that Lewis positive tumor cells may have an advantage in penetrating secondary organs because, like leukocytes, they can specifically adhere to activated vascular endothelia.

Die Deckung von grosflachigen Defekten an der Thoraxwand stellt zum einen haufig gesteigerte Anspruche an plastisch-chirurgische Methoden. Zum anderen entstehen derartige Defekte oft im Rahmen der Behandlung durch andere Disziplinen, insbesondere der Herzchirurgie und der Thoraxchirurgie. Ein interdisziplinares Vorgehen bietet sich daher an.

Development of metastases in cancer patients is usually due to tumour cells seeding from the primary. These disseminated cells can be detected in blood or bone marrow with the aid of immunocytochemical methods. The clinical significance of this is, however, still controversial. With the aim of determining the prognostic value of disseminated tumour cells, a meta-analysis of 20 publications reporting results based on data collected from a total of 2494 breast cancer patients, was carried out. The outcome of the meta-analysis was that 14 of the 20 studies provided evidence suggesting positive bone marrow findings to be an unfavorable prognostic factor. Five out of twelve studies provided multivariate analytic evidence of an impact of bone marrow status on recurrence-free survival. In five out of twelve studies, a univariate analysis showed a positive bone marrow finding to correlate significantly with overall survival, while a multivariate analysis found such a correlation in only two out of six studies. Owing to a lack of standardization, the results of the individual studies are not directly comparable. Against this background it is too early to say whether the detection of disseminated tumour cells in the bone marrow can be used as a prognostic factor with an impact on therapeutic strategies.

The present prospective study evaluates the impact of disseminated tumor cells in bone marrow of breast cancer patients on overall survival. In the whole patient cohort consisting of 1045 stage I – III patients the lymph node status was demonstrated to be the strongest independent prognostic factor, whilst no relevant impact of the bone marrow status could be demonstrated. Evaluating the node-negative (n = 596) and the node-positive patient (n = 439) cohorts separately revealed that the bone marrow status is an independent predictor for poor survival in node-positive patients (RR 1.59; CI 1.07 – 2.37; median follow up of 53 months).

PURPOSE In 1997, the immunocytologic detection of isolated tumor cells in bone marrow, termed micrometastasis, will be optionally included in the tumor-node-metastasis (TNM) classification indicated M1(i). In the present meta-analyses, 20 studies, which included 2,494 patients, regarding the prognostic influence of a positive bone marrow micrometastases (BMM) status on relapse-free and/or overall survival were analyzed. MATERIALS AND METHODS The literature search included the Medline and Current Contents bibliographic data bases from August 1980 to June 1997. The statistical evaluation considered the prognostic influence of the prevalence of micrometastatic cells in bone marrow on relapse-free and/or overall survival. The comparable effect estimate and its corresponding 95% confidence interval (CI) were calculated with the Mantel-Haenszel method using the originally published data of the retrieved studies. RESULTS The presence of epithelial cells in bone marrow was detectable in all carcinoma types, with a median prevalence of approximately 35%. Fourteen of 20 studies found a positive correlation between positive BMM status and reduced relapse-free survival by univariate analysis, but only five of 11 studies confirmed positive BMM status as an independent predictor of short disease-free survival. Regarding overall survival, positive BMM status was identified univariately in five of 12 studies, but multivariately in only two studies, as an independent factor of poor survival. Despite the heterogeneity of the studies, calculation of the relative risk (RR) for reduced relapse-free survival was possible for breast cancer, which resulted in a Mantel-Haenszel RR (RR(MH)) of 1.34 (95% CI, 1.27 to 1.42). CONCLUSION In conclusion, the results suggest that the prognostic impact of epithelial cells in bone marrow remains to be substantiated by further studies using standardized methodic protocols before its entrance in the TNM classification.

Seventy-eight transfusions of autologous platelets were given to eight alloimmunized patients receiving curative chemotherapy for acute leukemia. Platelets were collected at regeneration of hematopoiesis after a chemotherapy cycle, cryopreserved with 5% dimethylsulfoxide in liquid nitrogen, and retransfused during bone marrow aplasia following the next treatment cycle. The in vitro platelet recovery after freezing, thawing, and washing was 85 +/- 4%. The in vivo corrected count increment 1 h after autologous platelet transfusions was 11 +/- 5 x 10(9)/l. With the exception of moderate urticaria and slight nausea each after one transfusion, no immediate or chronic side effects occurred. The bleeding time was shortened and hemorrhage during bone marrow aplasia was prevented in all alloimmunized patients by autologous platelet transfusions.

Individual disseminated epithelial tumour cells were detected in bone marrow aspirates in 41 of 108 patients (37%) with squamous cell cancer of the head and neck region by an immunocytochemical technique based on monoclonal antibodies raised against the cytokeratin No. 19. In the clinical stage I (T1N0M0) tumour cells were detected only in 26.3% of the patients, whereas in stage IV (T4N0M0, T(all)N2-3M0, T(all)N(all)M1) almost twice as many patients (47.7%) presented with tumour cells in the bone marrow. Apparently, grade of differentiation of the tumour (grading) had no influence on the spread of single tumour cells. An influence of the different localisations of the primary tumour on tumour cell spread or the rate of tumour recurrence cannot as yet be discovered. Cytokeratin No. 19 expressing cells were not detectable in the bone marrow of 18 patients with non-malignant disease. Seventy-three patients were included in a follow-up study with a mean observation time of 25 months (range: 4-52 months). The presence of epithelial cells at the time of primary treatment appears to indicate a significantly higher risk of development of local or distant tumour recurrences (p = 0.01). Of 46 patients initially exhibiting no tumour cells in the bone marrow, only 14 had a clinical recurrence. Whereas 17 of 27 patients who presented with tumour cells in the bone marrow developed either a local tumour recurrence or distant metastases in different organs. Patients presenting with bone marrow tumour cells showed a significantly shorter disease-free survival than those without (p = 0.002).(ABSTRACT TRUNCATED AT 250 WORDS)

Expression of the epithelial cell adhesion molecule E-cadherin in primary and metastatic gastric carcinoma was examined using immunohistochemical analyses. Compared to normal mucosa, 92% of the primary tumors (n = 60) showed reduced E-cadherin expression, suggesting that down-regulation of this cell adhesion molecule is a common early event in gastric tumorigenesis. No significant correlation was found between E-cadherin expression and tumor diameter, lymphatic vessel invasion, Borrmann classification, lymph node status, or manifest metastases. Although advanced tumors (tumor stage 3/4) showed a loss of E-cadherin-positive cells (or = 50% cells/lesion, P = 0.0168), the most significant correlation was observed between low E-cadherin expression and cellular dedifferentiation (grading 3/4, P = 0.0001) and disintegration of tissue architecture (Lauren and WHO classifications, P = 0.0001). Low E-cadherin expression (or = 50% cells/lesion) was associated with tumor recurrence (P = 0.0013) and mortality (P = 0.0246). E-cadherin expression in metastatic lesions (n = 58) also correlated with the degree of glandular differentiation (P = 0.0001). Significant correlation (rs = 0.686) was observed between E-cadherin expression in primary and metastatic lesions from individual patients (n = 39). However, while metastases derived from E-cadherin-negative tumors remained negative, those originating from E-cadherin-positive tumors frequently demonstrated increased levels of expression. Evaluation of multiple metastases in 11 patients revealed uniformly strong E-cadherin expression in liver metastases, suggesting a possible regulatory role of the microenvironment.

Autologous platelets of 5 alloimmunized patients with acute leukemia in remission were cryopreserved with 5% dimethylsulfoxide in liquid nitrogen and retransfused in the following therapy-induced thrombocytopenic phase. The mean platelet recovery after freezing, thawing and washing was 85 +/- 6%. The mean corrected 1-hour increment in platelet counts was 11 (4-27) x 10(9)/l, i.e. 61% in comparison with fresh platelet transfusions in other patients. In vivo function of frozen platelets was documented by improvement of the posttransfusion bleeding time. Cryopreserved autologous platelets function hemostatically and can be used even for completely refractory patients and thus permit curative antileukemic therapy.

In node-negative breast cancer some 30% of patients will suffer a tumor relapse, despite primary curative therapeutic intervention. An adjuvant therapy is therefore in discussion, especially in presence of risk factors. Immunocytochemical staining of cytokeratin-positive cells in bone marrow aspirates with monoclonal antibodies offers a chance to detect tumorcell dissemination at an early stage of single tumor cells. In 103 consecutive patients 50% showed a positive bone marrow aspirate, in node negative patients still 34%. Based on the literature and tumor biology this method may be valid in selecting node-negative breast cancer patients for adjuvant therapy. In a prospective multi-center trial the prognostic value will be evaluated in regard to other prognostic factors. Additionally the benefit of an adjuvant therapy will be studied in a randomized prospective trial.

An astoundingly high frequency of micrometastatic cells have been found in bone marrow aspirates of patients with colon carcinomas (G. Schlimok et al., J. Clin. Oncol., 8:831-837, 1990), although these tumors very rarely metastasize to the skeleton. This observation has raised questions about the malignant potential of such cells. In a first attempt to characterize this potential, we have assessed the expression of major histocompatibility complex (MHC) class I antigens on bone marrow micrometastases, inasmuch as down-regulation of these molecules is a potential mechanism to escape from MHC class I-restricted lysis by cytotoxic T-cells. The two groups of cancer patients compared were those with tumors known to rarely (stomach and colon cancer) or frequently (breast cancer) manifest skeleton metastases. Bone marrow aspirates taken from these patients were probed for individual disseminated tumor cells using the immunoalkaline phosphatase technique with monoclonal antibody CK2 to the epithelial differentiation antigen cytokeratin 18 (CK-18), as described previously (G. Schlimok et al., Proc. Natl. Acad. Sci. USA, 84:8672-8676, 1987). Specimens containing CK18-positive cells were colabeled with monoclonal antibody W6/32 directed to a framework (or nonpolymorphic) antigenic determinant of MHC class I heavy chains associated with beta 2-microglobulin. W6/32-positive CK-18-positive cells could be detected in 25 of 54 patients (46.3%) with significantly higher incidences in 26 breast cancer patients (61.9%) as compared to 28 patients with carcinomas of the stomach and colon (27.3 and 29.4%). Independent from the origin of the primary carcinoma, the incidence of W6/32-negative CK18-positive cells was positively correlated to both the differentiation grade of the primary tumor (P less than 0.05) and appeared to be linked to the occurrence of regional lymph node metastases (statistically not significant) determined by conventional histological examination. The present results demonstrate for the first time that down-regulation of MHC expression on individual micrometastatic cells correlates to the differential pattern of metastasis obtained by comparing breast and gastrointestinal carcinomas. This finding together with the suggestive link to clinical risk factors supports the significance of reduced MHC class I expression for the survival of residual metastatic cells which is a major determinant of prognosis for patients with solid tumors.

Die fruhe Phase der Mikrometastasierung im Sinne disseminierter Einzelzellen last sich im Knochenmark als einem verhaltnismasig leicht zuganglichen und in den Kreislauf der hamatogenen Metastasierung eingeschalteten Organsystem nachweisen. Intraoperativ durch Beckenkammpunktion gewonnenes Knochen-marksaspirat wird uber einen FICOLL-Gradienten separiert, zu Cytospin-Praparaten aufgearbeitet und in einer APAAP-Farbung (Alkalische Phosphatase-Anti-Alkalische Phosphatase) untersucht.