Cancer radioimmunotherapy with alpha-emitting nuclides

In lymphoid malignancies and in certain solid cancers such as medullary thyroid carcinoma, somewhat mixed success has been achieved when applying radioimmunotherapy (RIT) with -emitters for the treatment of refractory cases. The development of novel RIT with -emitters has created new opportunities and theoretical advantages due to the high linear energy transfer (LET) and the short path length in biological tissue of -particles. These physical properties offer the prospect of achieving selective tumoural cell killing. Thus, RIT with -emitters appears particularly suited for the elimination of circulating single cells or cell clusters or for the treatment of micrometastases at an early stage. However, to avoid non-specific irradiation of healthy tissues, it is necessary to identify accessible tumoural targets easily and rapidly. For this purpose, a small number of -emitters have been investigated, among which only a few have been used for in vivo preclinical studies. Another problem is the availability and cost of these radionuclides; for instance, the low cost and the development of a reliable actinium-225/bismuth-213 generator were probably determining elements in the choice of bismuth-213 in the only human trial of RIT with an -emitter. This article reviews the literature concerning monoclonal antibodies radiolabelled with -emitters that have been developed for possible RIT in cancer patients. The principal radio-immunoconjugates are considered, starting with physical and chemical properties of -emitters, their mode of production, the possibilities and difficulties of labelling, in vitro studies and finally, when available, in vivo preclinical and clinical studies.     

A pragmatic approach to metal artifact reduction in CT: merging of metal artifact reduced images

The purpose of this study was to improve metal artifact reduction (MAR) in X-ray computed tomography (CT) by the combination of two artifact reduction methods. The presented method constitutes an image-based weighted superposition of images processed with two known methods for MAR: linear interpolation of reprojected metal traces (LI) and multi-dimensional adaptive filtering of the raw data (MAF). Two weighting concepts were realized that take into account mean distances of image points from metal objects or additional directional components. Artifact reduction on patient data from the jaw and the hip region shows that although the application of only one of the MAR algorithms can already improve image quality, these methods have specific drawbacks. While MAF does not correct corrupted CT values, LI often introduces secondary artifacts. The corrective impact of the merging algorithm is almost always superior to the application of only one of the methods. The results obtained with directional weighting are equal to or in many cases better than those of the distance weighting scheme. Merging combines the advantages of two fundamentally different approaches to artifact reduction and can improve the quality of images that are affected by metal artifacts.Symbols LI linear interpolation algorithm - MAF multi-dimensional adaptive filtering algorithm - MAR metal artifact reduction - f(x, y) object function at (x, y) - F(), F() normalized filter function in the direction of the coordinates and - I primary X-ray intensity - I₀ transmitted X-ray intensity - N_D number of detectors per detector row - p attenuation - p(,), p(,) projection data (fan and parallel geometry) - p_th lower threshold of the attenuation of adaptive filtering - p_max maximum value of the attenuation of adaptive filtering - (x, y) image point - projection angle in fan geometry - angle within the fan relative to the central ray - , , z, , minimum functions for adaptive filtering - projection angle in parallel geometry - orthogonal distance of a ray to the center of rotation in parallel geometry     

γ-Glutamyltransferase test as a new tool for identification of seminal stains

Summary A simple qualitative method for identification of seminal stains based on a high activity of -glutamyltransferase (-GTP) in human semen is described. It employs the release of -naphthylamine from N--glutamyl--naphthylamide by the -GTP action; -naphthylamine couples with Fast Garnet GBC salt to produce a strong brownish-red color. The data on its simplicity, specificity, and stability show that the present method is suitable for medicolegal examination of seminal stains as a preliminary test.     

Tunnel enlargement and changes in synovial fluid cytokine profile following anterior cruciate ligament reconstruction with patellar tendon and hamstring tendon autografts

There is growing evidence that cytokines such as tumor necrosis factor (TNF) , interleukin (IL) 1, IL-6, bone morphogenetic proteins (BMP), and nitric oxide (NO) play an important role in the pathogenesis of bone tunnel enlargement following anterior cruciate ligament (ACL) reconstruction. Furthermore, the release of these mediators has been considered a possible reason for the higher incidence of bone tunnel enlargement following hamstring tendon (HST) than following patellar tendon (PT) ACL reconstruction observed in several studies. In this investigation synovial fluid samples from 13 patients were collected immediately before (24±7 days after ACL rupture) and 7 days after ACL surgery and values of TNF-, IL-1, IL-6, NO, and BMP-2 were analyzed. Furthermore, the incidence of bone tunnel enlargement was assessed using radiographs 38±7 weeks after surgery. Six patients underwent autologous HST ACL reconstruction, and in seven patients an PT autograft was used. In the overall patient population there were significantly higher synovial fluid concentrations of IL-6 and BMP-2 postoperatively than preoperatively; TNF- showed a trend towards lower postoperative levels while IL-1 and NO remained unchanged. The concentrations of NO, TNF-, and IL-6 found in the present study were clearly higher than normal values given in the literature. Assessment of bone tunnel enlargement revealed an average increase in tibial tunnel width of 28.4±3.1% with comparable values for HST and PT ACL reconstructions. There was no significant correlation between bone tunnel enlargement and postoperative synovial fluid concentrations of TNF-, IL-1, IL-6, NO, and BMP-2. However, all patients with bone tunnel enlargement had higher postoperative concentrations of TNF-, IL-6, and NO in the synovial fluid. There were no significant differences in concentrations between HST and PT groups. In conclusion, we observed an association between tibial bone tunnel enlargement and elevated synovial fluid concentrations of IL-6, TNF-, and NO 7 days after ACL surgery indicating the potential involvement of these biological mediators in the pathogenesis of bone tunnel enlargement. However, there was no difference between HST and PT ACL reconstructions regarding synovial fluid contents of IL-6, TNF-, IL-1, NO, and BMP-2, suggesting a comparable biological response between these autografts following their use in ACL reconstruction.     

Stability of α-[11C]methyl-l-tryptophan brain trapping in healthy male volunteers

Purpose The purpose of this study was to assess the reproducibility in healthy volunteers of -[¹¹C]methyl-l-tryptophan ([¹¹C]MT) brain trapping imaging with positron emission tomography (PET), using volumes of interest (VOIs) and voxel-based image analysis.Methods Six right-handed healthy male volunteers (34.3±10.9 years) with a negative family history for psychiatric disorders were scanned twice in the resting condition, 22±17 days apart. An unbiased semiautomatic segmentation of the brain was used to define VOIs. The trapping constant K* (ml g^–1 min^–1) for [¹¹C]MT was calculated for the whole brain and seven brain regions using the graphical method for irreversible tracers. In addition, parametric maps of K* were obtained from dynamic scans using the same method. Comparison of test and retest K* functional images was performed using SPM99. Students paired t statistic was applied for comparisons of [¹¹C]MT brain trapping in a priori selected VOIs.Results [¹¹C]MT brain trapping in VOIs showed a mean variability 2.6±1.8% (0.3–5%) for absolute and 1.5±2.1% (1.4–4.1%) for normalized K*. Intraclass correlations between test and retest conditions were 0.61±0.34 for absolute K* values and 0.73±0.20 for K* values normalized by global mean. SPM99 analysis using a height threshold of p=0.05 (two tailed) and an extent threshold of 100 voxels showed no significant differences between scans.Conclusion Rest measurements in healthy male volunteers of the trapping constant for [¹¹C]MT, using PET, appeared to be stable during an average interval of 3 weeks.     

Genetic polymorphism of desialyzed alpha2 HS-glycoprotein by ultrathin isoelectric focusing

Summary The genetically determined polymorphism of ₂ HS-glycoprotein was analyzed by immunoblotting ultrathin-layer polyacrylamide gel isoelectric focusing in the pH range 4–6.5 and neuraminidase pretreated sera. In a Libyan population sample from Tripoli (n=110) three common phenotypes, ₂ HSG 1–1, 2–1, and 2–2, were observed. The allele frequencies were ₂ HSG¹=0.8364 and ₂ HSG²=0.1636. The theoretical exclusion rate in cases of disputed paternity is 11.8%.     

Population data of the HLA DQα locus in Dutch caucasians

Summary The HLA DQa amplification and typing kit has been designed to be used by the forensic community for purposes of identity testing. The introduction of any new DNA marker in forensic identity testing requires the establishment of a population database for the relevant population(s) [1]. To this end allele and genotype frequencies for the HLADQ locus were determined in a Dutch Caucasian population sample and compared with 7 other population genetic studies. In our population sample the HLA DQ genotype frequencies did not deviate from Hardy-Weinberg expectations and for this locus the power of discrimination is 0.94. A test for homogeneity of the HLA DQ population data based on the allele frequency counts for 8 Caucasian population samples was performed and significant differences were found (P = 0.007) . The differences in the frequency of the HLADQ 2 and 3 alleles are the major cause of this deviation. No deviation from population homogeneity was observed when we compared thegenotype frequency distributions among the 8 Caucasian population samples. Combined with the extensive validation studies from Comey and Budowle [7] and Helmuth et al. [8] this population genetic study will allow HLADQ typing to be used in forensic identity testing in the Netherlands.     

The dynamics of inflammatory cytokines in the healing process of mouse skin wound: A preliminary study for possible wound age determination

The dynamics of inflammatory cytokines such as interleukin-1 (IL-l, interleukin-1 (IL-1), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF) during the healing process of mouse skin wounds were examined using the enzyme-linked immunosorbent assay and immunostaining. The applicability of this examination for wound age estimation is discussed from the perspective of forensic pathology. After wound induction, mice were sacrificed at intervals ranging from 0 to 240 h. The levels of TNF and IL-1 began to elevate rapidly after wounding and reached a peak at 3 h. The IL-l level reached a peak at 6 h, and IL-6 peaked at 12 h. An infiltration of numerous leukocytes, indicatingacute inflammation, was observed at 3 and 6 h, and the main source of the cytokines was immunohistochemically identified as neutrophils. These results indicate that TNF and IL-1 play an important role in the commencement of inflammation. Rebound of cytokine levels, i.e. a re-increase, was observed at 72 h after wounding. Histological examination of the 72-h-old wound showed migration of fibroblasts and the formation of new granulation tissues, indicating the proliferative stage of the wound healing process. These experimental findings indicate that these cytokines have a close relationship to wound remodeling as well as to inflammation. From the viewpoint of forensic pathology, it is considered that inflammatory cytokines may become one of the markers for wound age estimation, but further studies are needed, especially those involving the investigation using human wound specimens with known time intervals after injury.This study was presented at the 79th Congress of the Medico-Legal Society of Japan (Yamagata, May 1995), the 16th Annual Meeting of the Japanese Society of Inflammation (Tokyo, July 1995) and the 74th Annual Meeting of the German Society of Legal Medicine (Aachen, September 1995).     

Accumulation of radioiodinated tyrosine derivatives in the adrenal medulla and in melanomas

Because tyrosine and dopa can be regarded as precursors of adrenomedullary hormones and melanin, radioiodinated derivatives of these compounds were tested for their accumulation in the adrenal medulla and in melanomas of various animal species. The highest level of accumulation in the adrenal medulla was attained in mice and rats with iodinated -hydroxy--methyltyramine, and in melanomas of mice with iodinated -methyltyrosine. The results could not be reproduced to the same extent in other species.     

Frequency of D1S80 and HLA DQα alleles in a Chinese population

Allele frequency distributions for the D1S80 (MCT118) and HLA DQ loci were determined in a Chinese population sample using the polymerase chain reaction (PCR). A total of 25 alleles and 100 phenotypes were observed for D 1 S80. The frequency of allele 18 was higher than allele 24 only in this Chinese population when compared to other reported populations. A total of 6 alleles and 21 possible phenotypes were observed for HLA DQ. The power of discrimination was 0.97 and 0.93 for D1S80 and HLA DQ, respectively.     

Vergleichende Untersuchungen über zeitabhängige Veränderungen im Eiweißspektrum von Vital- und Leichenblut mit der Celluloseacetatfolien-(CAF)-Elektrophorese

Zusammenfassung In systematischen Untersuchungen wird gezeigt, daß es mit zunehmender Hämolyse im Vital- und Leichenblut in der Elektrophorese auf Celluloseacetatfolien (CAF) zu einem Extragradienten im ₂-Globulinbereich kommt, der die relativen Verhältnisse im Elektropherogramm beeinflußt. Bei starker Hämolyse tritt eine Verschmelzung der ₂- und -Globulinbanden auf. Um eine diagnostische Beurteilung am Leichenblut mit der Elektrophorese durchzuführen, muß deshalb in erster Linie der Hämolysegrad berücksichtigt werden. Die durch Hämolyse bestimmten Veränderungen im elektrophoretischen Muster sind mit der Liegezeit am ehesten korreliert, wobei jedoch der Aussagewert bezüglich forensischer Zeitstellungen gering ist.     

Immunochemical studies on blood group H and B substances from human hair

Summary Blood group H and B substances were extracted from urea-treated human hair of group O and B individuals, respectively, with methanol-ethyl ether (1:1,v/v) and chloroform-methanol (1:1,v/v). The blood group activities of H and B substances were destroyed by H-decomposing enzyme (-l-fucosidase) fromBacillus fulminans and B-decomposing enzyme (-d-galactosidase) fromClostridium sporogenes Maebashi, respectively. It is concluded therefore that the extract from the hair of group O contained blood group H-active glycolipid with -l-fucose as the non-reducing sugar and the one from group B contained blood group B-active glycolipid with -d-galactose as the non-reducing sugar.     

‘Hot’ and ‘cold’ lesions detected by gallium-67 scintigraphy in a pyogenic liver abscess

Most pyogenic liver abscesses appear to be not in a ⁶⁷Ga-citrate study; rarely, a warm or cold area may also be seen. We present a patient with a pyogenic liver abscess that had both cold and hot areas in the ⁶⁷Ga-citrate study. This striking finding is probably related to the different stages of evolution of the abscess.     

Simultaneous Phenotyping of Genetic Markers for Paternity Testing

Summary Time- and cost-saving methods for paternity testing are described. Seventeen genetic systems were divided into six groups: (1) transferrin (Tf), factor B (Bf), and phosphoglucomutase 1 (PGM1); (2) group-specific component (Gc) or 1-antitrypsin (PI) and 2HS-glycoprotein (HSGA); (3) complement components C6 and C7, factor 13B (F13B), and plasminogen (PLG); (4) haptoglobin (Hp), C8 - chain (C81), and factor I (IF); (5) red cell acid phosphatase (ACP), esterase D (ESD), and glutamic-pyruvic transaminase (GPT); and (6) 6-phosphogluconate dehydrogenase (PGD) and glyoxalase I (GLO). Each group of systems was typed simultaneously by electrophoresis or isoelectric focusing (IEF) followed by staining or immunoblotting. These methods are very practical because they afford a considerable saving of time, work and expense, and facilitate semipermanent preservation of electrophoretic patterns.     

Detection of α S1-casein in vomit from bottle-fed babies by enzyme-linked immunosorbent assay

Summary This study describes a sensitive enzyme-linked immunosorbent assay (ELISA) using rabbit anti-bovine S1-casein antibody for the detection of commercial milk and milk-containing vomit. The antibody does not react with other human body fluids such as breast milk. The stability of S1-casein antigenic activity was examined after storage at different temperatures and enzyme digestion. There was no decrease after storage for one year at room temperature but 40% of the activity was lost after 6 months at 37°C. Enzyme digestion (6 hours, 37°C) resulted in 65 70% loss of activity but the antibody reacted with the peptide fragments of S1-casein. Vomit samples from 3 normal infants were tested by ELISA, and a S 1-casein could be detected in 1 cm² stain.     

The distribution of the HLA-DQα alleles and genotypes in the Finnish population as determined by the use of DNA amplification and allele specific oligonucleotides

Summary Allele and genotype frequencies for the HLA-130 locus were determined for use in forensic analyses and paternity tests in Finland. The polymerase chain reaction (PCR) and the reverse dot blot format were employed to detect 6 different HLA-DQ alleles. All 6 HLA-DQ alleles were detected among the 112 unrelated individuals with allele frequencies ranging from 5.8% to 32.6%. The distribution of the observed genotypes is in Hardy-Weinberg equilibrium. Additionally, this Finnish population sample is statistically similar to 2 other Caucasian sample populations. The power of discrimination of this system in the Finnish population sample is 0.92, suggesting this method may prove suitable for identification purposes.     

Disco-vertebral destructive lesions (So-called Andersson Lesions) associated with ankylosing spondylitis

Radiological examination of 31 patients with established ankylosing spondylitis revealed 58 disco-vertebral destructive lesions. Careful assessment of their radiological features permitted differentiation into two main types, (1) inflammatory and (2) non-inflammatory. The inflammatory type may represent either an active inflammatory process or a late sequel to such a lesion. In this series 44 lesions were included in this category, with a marked predilection for the lumbar spine. Moreover, all were observed within the first decade following the onset of the disease.The non-inflammatory type reflects an attempt of a stress fracture in the diseased spinal column to repair by connective tissue, causing a pseudoarthrosis. Eleven such lesions were obseved in this series all occurring more than 12 years after the onset of the disease. In a much smaller group of three cases-here termed the ambivalent type — differentiation between the inflammatory and non-inflammatory types could not be established on radiological grounds alone. It is of interest that all three cases occurred within the first year after the onset of the disease.It is suggested that the term Andersson lesion should be used to include all disco-vertebral destructive lesions related to ankylosing spondylitis—be they of inflammatory or non-inflammatory pathogenesis —as the better known term Romanus lesion includes all types of circumscribed marginal destructive lesions of vertebral bodies without involvement of the disc in ankylosing spondylitis.     

Impact of CT in the neurosurgical management of intracranial tumors

Summary A total of 103 brain tumor patients examined with CT, radionuclide brain scan, and angiography or pneumoencephalography, and all surgically verified, were studied to evaluate the impact of CT on the neurosurgical handling of brain tumors. CT alone was usually sufficient for optimal handling of astrocytoma patients, angiography in most meningioma cases, and pneumoencephalography in cases with sellar, suprasellar, and some other midline tumors. Information obtained only through CT sometimes altered the therapy. Sometimes it led to biopsy instead of a meaningless attempt at a radical excision; in other cases it permitted a radical excision otherwise not possible.     

Synovial fluid cytokine concentrations as possible prognostic indicators in the ACL-deficient knee

Approximately 44% of patients develop osteoarthritis (OA) following rupture of the anterior cruciate ligament (ACL) if the injury is left unrepaired. Restoring knee stability through reconstruction, while providing symptomatic relief, has not been shown to reduce the incidence of degenerative changes. In fact, recent studies have shown that 50%–60% of ACL-reconstructed patients go on to develop degenerative changes or frank osteo-arthritis. In light of these data, our group suggests that the cause of post-traumatic osteoarthritis is not biomechanical but biochemical. To test this hypothesis, we measured levels of nine cytokines which are important in modulating physiological and pathophysiological metabolism of cartilage in knee joint synovial fluid following ACL rupture. Our patient population contained both acute und chronic ACL ruptures. A total of 84 samples were collected and analyzed by enzyme-linked immunosorbent assay. On the basis of the data collected, we were able to identify subgroups of patients who, on the basis of their synovial fluid cytokine profile, may be at greater or leasser risk of developing post-traumatic OA. In general, patients displayed concentrations of interleukin-1 (IL-1), basic fibroblastic growth factor (bFGF), transforming growth factor- (TGF-), granulocyte/macrophage-colony stimulating factor (GM-CSF), IL-6, and IL-8 that we interpreted as being consistent with an inflammatory reaction. Of great interest is the fact that the levels of these cytokines were very similar in patients 4 weeks after injury and in chronic patients, leading us to hypothesize that a chronic smoldering inflammatory reaction persists after resolution of the acute effusion. This chronic inflammatory reaction could be the cause of cartilage degeneration seen in patients after ACL reconstruction. Furthermore, this study identified two distinct subpopulations of patients with dramatically differing levels of IL-1 and TNF- in their synovial fluids. IL-1 and TNF- were either high acutely and decreased to moderate levels over time, or they were zero acutely and remained zero or low. This is likely to be a very important observation as these two cytokines are key components of the inflammatory response and are the main intra-articular cytokines that provoke cartilage loss. It is our hypothesis that the group with high levels of IL-1 and TNF- contains the 50%–60% of the patient population which goes on to develop OA, despite knee stabilization with ACL reconstruction. Conversely, we suggest that the group whose synovial fluid contains no IL-1 or TNF- is protected from developing OA post-traumatically. IL-1-receptor antagonist protein (IRAP), the cytokine that antagonizes the biological effects of IL-1, was present acutely in our patient population; however, it was undetectable in chronic ACL-deficient knees. These data suggest the loss with time of an important cartilage-protective cytokine. The lack of IRAP chronically may allow IL-1 to act in an unrestrained manner resulting in cartilage loss. In addition, IL-1 promotes the secretion of many other inflammatory cytokines which may result in the chronic smoldering inflammatory reaction referred to above. These data encourage further study of the evaluation of cytokine concentrations in the ACL-deficient joint as a means of obtaining both prognostic and mechanistic information of clinical value to the physician.     

Biodistribution of iodine-125 tyramine transforming growth factor α antisense oligonucleotide in athymic mice with a human mammary tumour xenograft following intratumoral injection

The Watson-Crick base pairing rule provides the underlying principle for the antisense (AS) approach to inhibiting gene expression. Transforming growth factor (TGF) was the first growth factor to be associated with tumorigenesis, thus making the TGF (mRNA) a potential target for AS therapy and offering the potential for monitoring of the progression of malignancy by non-invasive imaging with radiolabelled AS phosphodiester. Probe labelling and biodistribution were studied in the present report. A 23-mer oligonucleotide sequence was synthesized and grafted in 5 with a tyramine group which was further radioiodinated. The radiolabelled AS was injected intratumorally in mammary tumour-bearing BALB/c mice (3 weeks after inoculation of 7·10⁶ NS2T2A mammary cells). Biodistribution was monitored by sequential scintigraphy and organ radioactivity after autopsy. The 5 tyramine group allowed specific and stable radiolabelling of the AS with¹²⁵I. The¹²⁵I AS oligonucleotide was rapidly cleared from the tumour by intestine and kidneys. Four hours after intratumoral injection, 6.5%±1.5% of the dose was retained in the tumour as non-degraded¹²⁵I AS. It is concluded that 5 tyraminylated AS provides information on the biodistribution of AS oligonucleotide following intratumoral injection. These data will contribute to the pharmacology of AS oligonucleotides which can be used for therapy.