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Plasma and red cell folate mean contents have been found to be significantly lower in 41 symptom-free beta-thalassaemia heterozygotes than in 21 controls. Such decreases must be considered as effects of an increased folate utilization caused by the enhanced total, both effective and ineffective, erythropoiesis. Since no close correlation has been found between packed cell volume and plasma or red blood cell folate levels, it seems that in 'healthy' beta-thalassaemia subjects, the degree of anaemia is not influenced by folate body reserves. However, the frequent finding of reduced red cell folate contents suggests that further folate imbalance might lead to clinically significant degrees of folate deficiency.

A vast number of substances have been suggested as possibly contributing to perturbation of the endocrine system. Several have been tested with different approaches ranging from yeast expression system of human oestrogenic receptors to human breast cancer cells assays. Surprisingly, no inhibition-binding experiments to steroid receptors on healthy human tissue have been performed so far. Our study provides inhibition binding experiments to oestrogens, progesterone, testosterone and retinoic acid receptors in prostate and uterine human tissue of organochlorine pesticides, phthalate esters, oestrogenic constituents derived from plants and phenol derivates. Affinities of significant extent of phthalates on oestrogenic, progestinic and androgenic receptors have not been detected. As for retinoic acid receptors, mono(2-ethylexyl)phthalate provokes a notable reduction of the binding of the tritiated retinoic acid, phtalic acid ethyl-n-butyl ester and 4-octylphenol show an affinity comparable to that of isoflavonoid genistein, whereas 4-nonylphenol reduces the binding of retinoic acid in prostate.

Objective: To compare euploid and aneuploid pregnancies with respect to maternal serum and amniotic fluid (AF) levels of the components of the plasminogen system. Methods: The study population consisted of 123 single pregnancies at the 17th gestational week, 16 with minor chromosomal abnormalities, 15 aneuploid, and 92 euploid. Results: Both groups with chromosomal abnormalities had significantly higher serum levels of urokinase plasminogen activator and its complexed form with its type-1 inhibitor compared with euploid pregnancies. In AF, tissue plasminogen activator was significantly lower in the aneuploid than the euploid group, whereas type-1 inhibitor of plasminogen activator was significantly higher in the cases with minor chromosomal abnormalities compared with euploid. At cutoff levels set at 100% sensitivity, the complexed form of urokinase plasminogen activator with its type-1 inhibitor had the strongest specificity (66.3%); after logarithmic transformation, its serum level was 7.53 times higher in aneuploidies than euploidies. Conclusion: Aneuploid pregnancies appear to be accompanied by abnormalities of the plasminogen activation system, which could lead to impaired placental perfusion and thus to abortion, fetal death, and fetal growth restriction.

A new efficient approach for the preparation of (188)Re radiopharmaceuticals starting from [(188)ReO(4)](-), produced at a carrier-free level through the (188)W/(188)Re generator system, is described. The reaction procedure was based on the combined action of different reagents and has been applied in detail to the preparation of the therapeutic agent (188)Re(V)-DMSA (H(2)DMSA [meso-2,3-dimercaptosuccinic acid]). The most efficient combination required the use of SnCl(2), oxalate ions, and gamma-cyclodextrin. These were reacted with [(188)ReO(4)](-) and H(2)DMSA to afford the final radiopharmaceutical in high radiochemical purity, at room temperature, and in weakly acidic solution. The role played by the various reagents in the reaction was investigated. It was found that SnCl(2) behaved as the actual reducing agent, whereas oxalate and gamma-cyclodextrin greatly enhanced the ease of reduction of [(188)ReO(4)](-) through the action of two hypothetical mechanisms. In the first step of the reaction, oxalate ions gave rise to the formation of Re(VII) complexes with the concomitant expansion of the coordination sphere of the metal. This process strongly favored the electron transfer between Sn(2+) and Re(+7) centers, giving rise to intermediate reduced rhenium complexes. These species were further stabilized by the formation of transient host-guest aggregates with gamma-cyclodextrin and finally converted into (188)Re(V)-DMSA through simple replacement of the coordinated ligands by H(2)DMSA.

The trigeminovascular system is considered to play a role in the mechanism of migraine headache. Novel in vitro animal models that investigate the release of neuropeptides may be of help to understand the pathophysiology and pharmacology of trigeminal neurons. Here, we examined the release of the immunoreactivity (LI) of the sensory neuropeptides calcitonin gene-related peptide (CGRP) and substance P (SP) from slices of rat and guinea pig trigeminal ganglia with proximal nerve trunks attached. Electrical field stimulation (EFS, 10 Hz), high K+ medium (50 mM) and capsaicin (1 μM) caused a significant increase in CGRP-LI outflow. SP-LI was also released after exposure to EFS, high K+ and capsaicin. The increase in CGRP-LI outflow induced by EFS was markedly reduced in a Ca2+-free medium and by pretreatement with a high capsaicin concentration, tetrodotoxin, ω-conotoxin, dihydroergotamine and sumatriptan. Sensory neuropeptide release from slices of rat trigeminal ganglia with nerve trunks attached fulfills the criteria required to define it as a neurosecretory event. This is a novel method for studying trigeminal neuron pathophysiology and the action of antimigraine drugs.

A multicentre survey of the quality control of 99 Tc m generators has been completed : 245 generators from seven different commercial sources were tested over a period of 2 years. The results indicate that the mean pH of the eluates was 5.8 ± 0.6; the aluminium contents were typically < 10 ppm; the radiochemical purity was 99.8 ± 0.4% and the median 99 Mo content was 3.8 × 10 -4 percent. The elution profiles gave a volume of 1.9 ml to obtain 50% of the total eluted activity and of 4.9 ml to obtain 95%. Other radionuclide impurities and heavy metal breakthrough were evaluated by graphite furnace absorption spectrometry and inductively coupled plasma mass spectrometry. National guidelines for the standardization of radiopharmacy procedures are currently being compiled.

The synthesis, at tracer level, of two Tc-99m complexes having the same chemical composition and structure, but differing by one electron in the total electron counting, is reported. These compounds have been prepared by reacting [99mTcO4]− with the piperidinium salt of the ligand ferrocenedithiocarboxylate {[Fe(II)(C5H4CS2)(C5H5)]− = FcCS}, in the presence of N-methyl S-methyldithiocarbazate as donor of N3− groups, and triphenylphosphine or SnCl2 as reducing agents. The formation of the neutral complex [99mTc(N)(FcCS)2] (compound A) and of the monocationic, mixed-valence complex [99mTc(N)(FcCS) (FcCS∗)]+ (compound B) {FcCS∗ = [Fe(III)(C5H4CS2)(C5H5)]} was obtained in high yield. Both complexes comprise a terminal Tc ≡ N multiple bond and two FcCS ligands coordinated to the metal center through the two sulfur atoms of the -CS2 group, but they differ in the oxidation state of one of the two iron atoms of the coordinated FcCS ligands. In complex A, the two Fe atoms are both in the +2 oxidation state, while in B, one Fe atom is in the +2 and the other is in the +3 oxidation state. Thus, B is a mixed-valence Fe(II)-Fe(III) complex. B is easily converted into A by one-electron exchange with various reductants such as triphenylphosphine and excess SnCl2. Biodistribution studies in rats showed that complexes A and B are mostly retained in lungs and liver without any significant uptake in organs such as heart and brain.

Summary We used brain single-photon emission tomography (SPET) to detect hypoperfused areas in 15 consecutive Behcet syndrome patients. Five were suspected of having neuro-Behcet's syndrome, having at least one neurological symptom. For these patients, SPET was performed within 1 month of the onset of nervous system involvement. The 15 patients fulfilled the criteria of the International Study Group for Behcet syndrome. Neurological assessment and SPET were complemented by EEG in all five patients with suspected neuro-Behcet syndrome and by magnetic resonance imaging in three. Brain SPET detected hypoperfused regions in all five neurological patients; EEG showed abnormalities in three. Magnetic resonance imaging was normal in the three patients in whom it was performed. SPET was negative in all patients without neurological involvement and 20 healthy controls. SPET detected a reduction in brain blood flow in early neuro-Behcet syndrome, but there was no definitive correlation between the hypoperfused brain regions and the clinical features. Further studies are required to evaluate the significance of brain hypoperfusion and the value of SPET in the early diagnosis of neuro-Behcet syndrome. (© 1198 Lippincott Williams & Wilkins)

Carnitine derivatives, such as propionyl-l-carnitine (PLC), have been shown to improve walking distance in patients with obstructive peripheral artery disease (PAOD). The aim of this study was to ascertain whether technetium-99m sestamibi leg scintigraphy may be a useful tool in the evaluation of changes in skeletal muscle metabolism induced by chronic therapy with PLC. Twenty patients with clinical and instrumental evidence of PAOD were randomly assigned to a 3-month period of therapy with either PLC or placebo. Rest 99mTc-sestamibi leg scintigraphy and echo-Doppler sonography were performed on all subjects immediately before and upon completion of the treatment period. At the end of the protocol the following results were observed in patients who underwent PLC administration: (a) a significant increase in both thigh and calf 99mTc-sestamibi uptake, in comparison with baseline values (P0.001); (b) the absence of statistically significant modifications of Doppler blood flow indices of the lower limbs. In conclusion, after chronic administration of PLC, a significant increment in skeletal muscle uptake of 99mTc-sestamibi was demonstrated without any apparent change in regional blood flow. This fact, if proven in further studies, may suggest a role for this tracer as a non-invasive probe of tissue bioenergetics.

Ras/p21 oncoprotein expression and K-ras mutations were analysed by Western blot and/or K-ras oligonucleotide hybridization in 78 primary ovarian cancers, 20 omental metastases, two low malignant potential tumours (LMP), nine benign ovarian tumours and 10 normal ovaries. A cut-off value of an integral of absorbance (i.a.) of 2.20, obtained by receiver operating characteristic (ROC) curve, was shown to be the best cut-off for defining p21 positivity. p21 levels were higher in malignant tumours than in benign tumours (median 2.10 i.a. vs median 1.22 i.a.; P = 0.014) and in omental metastases than in primary ovarian carcinomas (median 2.54 i.a. vs median 2.1 i.a.; P = 0.0089). p21 overexpression did not correlate with any of the clinicopathological parameters examined. Follow-up data were available for 63 patients. A significant relationship was shown between p21 positivity and a shorter overall survival (OS) (P < 0.03) and progression-free survival (PFS) (P < 0.03). In multivariate analysis only the presence of ascites, p21 levels and epidermal growth factor receptor status retained an independent prognostic role. K-ras gene mutations were frequently detected in benign and low malignant potential tumours (71.4%), which were mostly mucinous (P = 0.0152). Images Figure 1 Figure 2

The serological tumor marker tissue polypeptide antigen (TPA) and the more recently identified tissue-specific polypeptide antigen (TPS) have been reported to be indicators of the proliferation rate of the tumor. In the present investigation we compared the cytosol level of the two markers with the proliferative activity of the tumor measured using the3H-thymidine labelling index. The preliminary results presented here show that higher TLI is associated with lower cytosol levels of both TPA and TPS. TPA and TPS in the cytosol were significantly associated. These findings are in agreement with the previously demonstrated association between high TPA cytosol levels and better prognosis in breast cancer. Further studies are ongoing in order to:1. confirm these findings in a larger patient series;2. investigate any possible prognostic indication provided by TPS;3. evaluate any possible biological meaning of the negative association between TPA/ TPS and TLI in the cytosol of breast cancer.

The use of tumor marker tests has increased progressively in the last decade concomitant with the advent of new monoclonal antibodies and their growing use in clinical oncology for various follow-up programs. External quality assessment (EQA) schemes widely adopted in clinical chemistry, have been extended in the last decade to immunoassays of hormones and tumor markers. EQA results can provide realistic information on the quality of the assays, performed under routine conditions. The goal of this article is to report the main results and discrepancies encountered so far in External Quality Assessment programs on tumor markers.

An in vitro system has been set up to study the possible genetic effects of diagnostic ultrasound (US) on the developing fetus. Amniotic cells were cultured by the in situ technique. Well established clones were exposed to US for various lengths of time using a linear array transducer within a sound transparent apparatus. Cells were then grown in the presence of 5-bromo-2'-deoxyuridine and processed for scoring the number of sister chromatid exchanges (SCE) as the cytogenetic endpoint. There was no linear relationship between the SCE frequency and the duration of US exposure. Variance analysis showed that only interindividual variability was a significant component of total variation. Neither the main effect of treatment nor the interaction effect were statistically significant. The data suggest that US delivered from a diagnostic unit to actively growing cultured fetal cells in a system closely mimicking the conditions of US exposure during amniocentesis does not induce SCE.

The ras-encoded p21 protein expression was investigated in 18 normal endometrial tissues and in 37 human primary endometrial carcinomas by Western blotting analysis. Scattered p21 levels were found in normal specimens (mean = 1.29 OD; median = 1.10 OD; range = 0.33-2.65). The p21 levels were significantly higher in secretory (mean = 1.99 OD; median = 2.16 OD; range = 0.71-2.65) than in proliferative (mean = 0.97 OD; median = 1.07 OD; range 0.38-1.73) endometrium (P = 0.009) and higher in primary endometrial carcinomas (mean = 2.05 OD; median = 2.04 OD; range 0.21-4.36) than in normal proliferative tissues (P = 0.004). Immunohistochemical analysis showed that most of the tumor cells expressed p21 oncoprotein while the stromal component was unreactive. No correlation between p21 expression and histopathological characteristics of the tumors was observed. Moreover, estrogen receptor (ER)-positive tumors expressed higher p21 levels than did ER-negative tumors (77% vs 33%; P = 0.009). A similar trend, although not statistically significant, was found between p21 values and progesterone receptor expression (74% vs 44%; P = 0.060). On the other side, p21 levels were unrelated to epidermal growth factor receptor levels. Further studies should verify the possible significance of p21 expression in the prognostic characterization of patients with endometrial cancer.

Mitogenic properties have been demonstratedin vitro for the lysosomal acidic protease cathepsin-D (cath-D). We investigated possible relationships between cath-D cytosol cell content and tumor proliferative activity in a series of 129 operable breast cancer patients. For total cytosol cath-D evaluation, a solid phase two-site immunoradiometric assay was utilized on tumor cell cytosol obtained for hormone receptor assay (DCC method). The percentage of S-phase cells was analyzed by 3H-thymidine autoradiographic assay. Median 3H-thymidine Labeling Index (3H-Tdr-LI) of the series was 2.7%; median cath-D content resulted 57 pmol/mg of protein cytosol and was significantly higher in node-positive with respect to the node-negative subgroup (p

Cathepsin-D is a lysosomal protease that can serve as an important prognostic cytosolic factor in breast cancer. To evaluate the performance of a commercially available immunoradiometric assay kit, 15 lyophilised cytosol participating EORTC laboratories for preparations containing various levels of cathepsin-D have been sent to 13 participating EORTC laboratories for quality control purposes. The between-laboratory variation, when expressed as coefficient of variation, did not exceed 24%. The within-laboratory variation was assessed by analysing five samples of the 15 specimens containing lyophilised material from a homogeneous cytosol pool. The mean within-laboratory coefficient of variation was 7%. Analysis of human breast tumour cytosols in one of the participating EORTC laboratories resulted in a median within-assay variation between duplicate measurements of 2.1% (n = 408), with a between-assay variation of a pooled human breast tumour cytosol of 4.6% (n = 11).

A total of 1095 patients with operable breast cancer and en-rolled in a randomized clinical trial were analysed for estrogen (ER) and progesterone (PgR) receptor content of their primary tumor, and the relationships between steroid receptor status and several epidemiologic characteristics were studied. The proportion of ER+ and median ER levels increased with age: compared to women younger than 40, those aged 66 or more were approximately three times more likely to have an ER+ tumor (OR = 3.0, 95% C.I. = 1.6–5.7). This difference tended to be more marked after comparison between patients with ER > 100 fmol/mg protein and ER- within the same age groups: OR = 7.04, 95 % C.I. = 2.89–17.12. No association emerged between age and PgR. ER status and concentrations were independent of menopausal status after adjustment for age, whereas the proportion of PgR+ and PgR levels were significantly lower in postmenopausal patients of the same age. The distribution of ER and PgR profiles was similar in relation to family history of breast cancer, reproductive events and other selected epidemiologic characteristics of the patients.

OBJECTIVE: To test the effect of ampicillin on amniotic interleukin-6 (IL-6) and prostaglandin E 2 (PGE 2 ) release. METHODS: In an in vitro study, IL-6 and PGE 2 release from amnion-like Wistar Institute Susan Hayflick cells was assayed under basal conditions, as well as after incubation with ampicillin. In an in vivo study, amniotic fluid IL-6 was assayed in a total of 212 patients submitted to genetic amniocentesis during the 17th week of their singleton physiological pregnancy. The study population was subdivided as follows: 92 patients sampled before ampicillin administration, 70 patients sampled 4 hours after administration of 1 g ampicillin, and 50 patients sampled 12 hours after administration of 1 g ampicillin. RESULTS: At doses ranging from 10 -7 to 10 -4 M, ampicillin decreased IL-6 release from Wistar Institute Susan Hayflick cells. The drug effect was already statistically significant (-30%; P

The authors have successfully built and characterised a small animal PET based on 4 rotating detectors with a spatial resolution

At Ferrara University the authors have built a dedicated 3-D PET scanner (YAPPET) with a very high spatial resolution and sensitivity as necessary for radiopharmaceutical studies on rats and mice. In collaboration with the Institute H.S. Raffaele in Milano the authors have performed both distribution and dynamical uptake studies of various radioactive tracers in rats. They have performed tomographic studies after injecting live rats with FDG and FESP labeled with /sup 18/F, with FE-CIT and Flumazenil labeled with /sup 11/C, and with /sup 18/F. The /sup 18/F-FDG, /sup 18/F-FESP and /sup 11/C-FE-CIT images show the different distribution of the various tracers in the rat brain. The /sup 18/F was injected for imaging the rat skeleton. In addition, the authors studied the uptake and washout curve of /sup 11/C Flumazenil in the rat brain from tomographic images.

At present, at the Dept of Experimental and Clinical Medicine of the University of Ferrara, the YAP-(S)PET scanner is employed in small animal SPECT studies, with 99mTc labelled radiotracers, dedicated to obtain quantitative activity measurements in a region of interest (ROI) directly from a reconstructed image of a rat, with respect to the traditional ex-vivo measurements of activity with gamma-counters. To achieve this goal, the scanner was calibrated referring it to a Isodose dose calibrator based on Geiger-Muller counters which previously underwent an absolute calibration with various certified activity sources. The calibration factor K was defined as the ratio between the activity measured with the Isodose (MBq) and the image count rate (cps). With the calibrated scanner we compared results of activity measurements from images of whole-rat heart acquisitions vs explanted heart. We find that whole-rat quantitative precision is within 3.2% considering 9% attenuation in the rat tissue

We report a case of a woman who came to our attention because of hypokalemia, hyperreninemia and hyperaldosteronemia but with normal blood pressure. Under suspicion of a normotensive renal artery stenosis captopril and baseline scintigraphies were performed. Captopril scintigraphy demonstrated a bilateral progressive retention of radiopharmaceutical without significant excretion. The baseline study revealed a complete normalization of the scintigraphic picture. A Magnetic Resonance Angiography (Angio-MRI) performed to evaluate renal arteries gave completely normal results. On the basis of the clinical picture and imaging findings a diagnosis of Bartter’s syndrome was formulated. Renal function in Bartter’s syndrome patients is maintained by hyperactivation of the renin angiotensin system. Acute administration of captopril in these patients induces an increase of renal plasma flow whereas it has no effects on glomerular filtration rate thus inducing a decrease of the filtration fraction: post captopril renal scintigraphy of our patient depicted exactly this feature. Although the diagnosis of Bartter’s syndrome is based on the clinical picture and biochemical abnormalities, scintigraphic tests could be useful in differentiating Bartter’s syndrome from other causes of hypokalemia.

Item does not contain fulltext Endogenous heterophilic antibodies in blood are known to interfere with two-site enzyme-linked immunosorbent assays (ELISAs) evoking false positive signals. In the present study, we describe an assay for the assessment of components of the plasminogen activation system (uPA, tPA and PAI-1, and their complexes) in blood which is not susceptible to interference by heterophilic antibodies. In the ELISA format, two avian (duck, chicken) antibodies are employed in the pre-analyte and two mammalian (rabbit, goat) antibodies in the post-analyte stage. The assay is compared to our earlier reported ELISA for measuring uPA, tPA and PAI-1 components in tumor tissue extracts. Applying the so-called "nonsense formats", designed against non-existent components, to the NIBSC reference preparation of rheumatoid factor (RF), no response was found with the new assay, whereas a clear RF dose-dependent interfering signal was observed with the original assay designed for tumor tissue extracts. Analysis of tumor-tissue based international reference preparations (RBG EORTC 101094 and 040297), human anti-mouse antibodies (HAMA) containing sera, and sera from patients with rheumatoid arthritis (RA), also displayed no false positive signals. In conclusion, we have developed an ELISA that permits the determination of blood levels of components in the urokinase system, free from disturbance by endogenous heterophilic antibodies.

Asymmetrical heterocomplexes containing a terminal technetium-nitrogen multiple bond coordinated to one diphosphine ligand (PNP) and one dithiocarbamate ligand (DBODC), were obtained through a simple two-step procedure under controlled conditions. The resulting complexes [99mTc(N)(PNP)(DBODC)]+ are monocationic, and possess a distorted square-pyramidal geometry where the Tc triple bond N multiple bond occupies an apical position and the diphosphine and dithiocarbamate ligands span the residual four coordination positions on the basal plane through the two phosphorus atoms and the two sulfur atoms, respectively. Biodistribution data in rats demonstrated that these complexes were rapidly extracted by the myocardium, and retained in this region for a prolonged time. After a few minutes post-injection, lung uptake became negligible, and liver washout was extremely rapid and quantitative. Analysis of heart/liver uptake ratios for these complexes revealed that their values increased exponentially in time, and after 60 min post-injection liver activity was almost completely eliminated into the intestine. Comparison with heart/liver ratios determined for 99mTc sestamibi and 99mTc tetrfosmin showed that values for these latter compounds were approximately 10 times lower than those measured for [99mTc(N)(PNP)(DBODC)]+ complexes at 60 min post-injection. In conclusion, the monocationic tracers [99mTc(N)(PNP)(DBODC)]+ exhibit high myocardial uptake in rats and dramatically high heart/lung and heart/liver ratios, suggesting that this novel class of perfusion agents could be conveniently employed to obtain heart images with superior imaging quality.

For the last 20 years, the most widely used method for assessment of the renin-angiotensin system has been the plasma renin activity assay (1). Although the assay gives useful information on the enzymatic function of the renin molecule, the intrinsic characteristics of this method limit its analytical accuracy (2). Immunoradiometric assays for renin, developed in 1985, overcame this limitation of the enzymatic assay because the well-defined monoclonal antibodies provided the means for direct quantification of specific active forms of the enzyme molecule (3)(4). Nevertheless, a critical debate has developed over the use of these assays (5)(6)(7)(8)(9)(10). A pilot study (11)(12) in a limited number of laboratories (eight Italian centers) found satisfactory indexes of precision for a direct immunoradiometric renin assay (Eria Diagnostics, Sanofi-Pasteur). In our previous experience, the inter- and intralaboratory reproducibility indices of this commercial immunoradiometric assay appeared to be better than those that had been achieved with an enzymatic assay (REN-CTK; Sorin Biomedica), with the differences probably being attributable to the greater complexity of the procedure for the latter assay (11). Various reports have been published during the past 30 years describing useful tools to monitor the performance of immunoassays …

A multicentre observational study was conducted by the Italian Association of Nuclear Medicine between 1996 and 1998. Twenty-nine Nuclear Medicine Departments participated. The aims of the study were to systematically evaluate the efficacy, toxicity and repeatability of radionuclide therapy of painful bone metastases (RTBM) in a large number of patients and to assess its incidence in patients with prostate cancer. Out of 818 treatments performed with a single i.v. dose of 148 MBq of strontium-89 chloride or 1,295 MBq of rhenium-186 hydroxyethylidene diphosphonate (HEDP), 610 could be evaluated (527 with 89Sr and 83 with 186Re-HEDP). Eighty-one patients received multiple (up to five) RTBM. The total number of retreatments was 100. Patients were followed up for a period of 3-24 months. Results, assessed according to pain relief and consumption of analgesic drugs, were expressed at four levels: 1, no response; 2, mild response; 3, good response; 4, excellent response. Responses were: level 1 in 19%, level 2 in 21.3%, level 3 in 33.3% and level 4 in 26.4% of cases. Retreatments showed significantly (P0.01) worse responses (48% levels 3+4), in comparison to first RTBM. Duration of palliation was 5.0+/-3.5 months, and was longer in cases of excellent response, in first RTBM, in patients with limited metastases and when 89Sr was used. Better responses were found in cases of limited skeletal disease, under good clinical conditions, when life expectancy exceeded 3 months, and in radiologically osteoblastic or mixed bone lesions. The only statistically significant predictive factor was life expectancy (P0.001). Flare phenomenon (14.1% of cases) did not correlate with the response. Haematological toxicity (mild to moderate in most cases) mainly affected platelets, and was observed in 25.5% of cases overall and in 38.9% of retreatments. RTBM did not seem to prolong life, though in some cases scintigraphic regression of bone metastases was observed. The two radiopharmaceuticals did not show any statistically significant differences in palliative efficacy and toxicity, either in first RTBM or in retreatments.

A new labeling approach for incorporating bioactive peptides into a technetium-99m coordination complex is described. This method exploits the chemical properties of the novel metal-nitrido fragment [99mTc(N)(PXP)]2+, composed of a terminal Tc[triple bond] N multiple bond bound to an ancillary diphosphine ligand (PXP). It will be shown that this basic, molecular building block easily forms in solution as the dichloride derivative [99mTc(N)(PXP)Cl2], and that this latter complex selectively reacts with monoanionic and dianionic, bidentate ligands (YZ) having soft, pi-donor coordinating atoms to afford asymmetrical nitrido heterocomplexes of the type [99mTc(N)(PXP)(YZ)]0/+ without removal of the basic motif [99mTc(N)(PXP)]2+. The reactions of the amino acid cysteine was studied in detail. It was found that cysteine readily coordinates to the metal fragment [99mTc(N)(PXP)]2+ either through the [NH2, S-] pair of donor atoms or, alternatively, through the [O-, S-] pair, to yield the corresponding asymmetrical complexes in very high specific activity. Thus, these results were conveniently employed to devise a new, efficient procedure for labeling short peptide sequences having a terminal cysteine group available for coordination to the [99mTc(N)(PXP)]2+ fragment. Examples of the application of this novel approach to the labeling of the short peptide ligand H-Arg-Gly-Asp-Cys-OH (H(2)1) and of the peptidomimetic derivative H-Cys-Val-2-Nal-Met-OH (H2) will be discussed.

The synthesis of a hybrid constituted by distamycin A and cysteine labeled with the γ-emitting radionuclide 99m Tc to afford the conjugate complex 5 is reported. This new radiopharmaceutical is of potential interest as tumor imaging agent in diagnostic nuclear medicine. The preparation of the hybrid distamycin A-cysteine 4 has been achieved by coupling deformyldistamycin A and Boc-Dmt-OH. Compound 4 was then successfully labeled with 99m Tc by reaction with the novel, high-electrophilic, metal-containing fragment [ 99m Tc(N)(PP)] 2+ (PP=diphosphine ligand) yielding the 1:1 complex 5 .

The octapeptide D-Ala-Ser-Thr-Thr-Thr-Asn-Tyr-Thr-NH 2 ([D-Ala 1 ]TNH 2 ), an analog of peptide T (H-Ala-Ser-Thr-Thr-Thr-Asn-Tyr-Thr-OH) associated with CD 4 /T 4 receptors involved in human immunodeficiency virus infection, was combined with the chelating polyazamacrocycle 1,4,8,11-tetraazacyclotetradecane (cyclam) to afford the bifunctional ligand cyc-[D-Ala 1 ]TNH 2 . This was then reacted with [ 99m TcO 4 ] − and Sn 2+ to yield the monocationic complex [ 99m Tc(O) 2 (cyc-[D-Ala 1 ]TNH 2 )] + . Biological activity of both the cyclam-peptide conjugate and the resulting Tc-99m complex were evaluated by measuring their chemotactic indexes. Results showed that N-cyclam acylation and subsequent labeling with Tc-99m of [D-Ala 1 ]TNH 2 were tolerated, and both cyc-[D-Ala 1 ]TNH 2 and [ 99m Tc(O) 2 (cyc-[D-Ala 1 ]TNH 2 )] + retained the high chemotactic capacity of the original octapeptide. Biodistribution of the Tc-99m complex was carried out in rats. Fast blood clearance and no accumulation in organs of interest were observed.

New Tc-99m radiopharmaceuticals with the ligand ( o -hydroxyphenyl)diphenylphosphine have been prepared and their biodistributions evaluated in rats. The monoxo Tc(V) complex [ 99m Tc(O)Cl(PO) 2 ], the Tc(IV) complex [ 99m Tc(OH) 2 (PO) 2 ], the Tc(III) complex [ 99m Tc(PO) 3 ], and the nitrido Tc(V) complex [ 99m Tc(N)(PO) 2 ] have been characterized by TLC and HPLC chromatography, and their chemical structure elucidated by comparison with the corresponding complexes obtained using the β-emitting isotope Tc-99g. Biodistribution studies of these complexes have been carried out in rats.

The diagnosis of central nervous system (CNS) involvement appears to be a major problem in systemic lupus erythematosus (SLE), especially when the clinical signs are non-specific or neuroimaging is unremarkable. Two SLE patients with mild neuropsychiatric manifestations were studied with magnetic resonance imaging (MRI), single photon emission tomography (SPET) and localized proton magnetic resonance spectroscopy (H-1 MRS). MRI was normal in both patients. SPET revealed areas of hypoperfusion in both patients. H-1 MRS demonstrated metabolic abnormalities in the regions corresponding to the hypoperfused areas. A correlation between H-1 MRS and SPET was noted: patients with mild neuropsychiatric SLE may have disturbances evident on SPET and H-1 MRS in the presence of normal anatomy on MRI, suggesting that CNS involvement in SLE has very strong physiological and neurometabolic components in individual patients.

Four different methods to measure in parallel the erbB2 protein expression (p185neu) were evaluated in order to: a) compare two enzyme immunoassays with the immunohistochemical assays (IHC) and western blotting (WB) and b) extrapolate eventual relationships between erbB2 and biological parameters. Tissue samples from 248 patients with primary breast cancer were consecutively assayed. We used two different cut-off levels for WB, ELISA, and EIA, defined as follows: 1) the highest level of expression of non malignant tissue was chosen as the discriminant threshold between 'low' and 'elevated' samples: 2) the elevated group was further subdivided into two subgroups: 'intermediate' and 'high', according to their median value. According to the first cut-off, the results were considered 'elevated' in about 52% of cases with the three biochemical methods, while using the second cut-off the percentage lowered to about 26%. Considering this cut-off, the concordance rates between the paired biochemical methods ranged between: 78.4% (WB vs EIA), 93% (ELISA vs EIA), and 82.6% (ELISA vs WB). The comparison between biochemical and immunohistochemical methods gave these concordance rates: 82% (WB vs IHC), 90.5% (ELISA vs IHC), and 85.5% (EIA vs. IHC). According to the first cut off level, 27.5% of tumor samples showed IHC detectable p185 levels, in agreement with other immunohistochemical studies. The relationship between high erbB2 and estrogen and progesterone receptors showed an inverse association. No relationship was found between erbB2 and axillary lymph node positivity or tumor size. In short, the results of the four methods seem generally well correlated; nevertheless, it appears that different methodological approaches of measuring p185neu are not completely equivalent, and there is a need for an authoritative standardization and quality control for clinical applications.

Data collected in the 1993 and 1994 cycles of an international external quality assessment (EQA) program and in a national multicenter collaborative study were cumulatively analyzed to evaluate the standardization of the methods currently in use for the assay of mucinous tumor markers CA 19-9, CA 15-3 and CA 125. On average the between-laboratory variability was 15.2 and 16.0 CV% for CA 15-3 and CA 125 respectively; the between-laboratory variability found for CA 19-9 was markedly worse (mean 28.3 CV%). The variability component attributable to systematic differences between different methods/kits was relatively small for CA 15-3 and CA 125 (18% and 24% of the total variability) but markedly larger for CA 19-9 (48% of the total variability). The agreement of CA 19-9 results worsened in the last few years when new nonisotopic techniques became available. The precision of the methods/kits most used in the survey ranged from 9.9 to 13.3 CV% for CA 125 and from 11.6 to 13.9 CV% for CA 15-3. For these two tumor markers the precision of the traditional IRMAs does not appear different from that of the new fully automated nonisotopic techniques. The precision of CA 19-9 methods was on average worse (from 11.7 to 19.6 CV%) although two automated systems exhibited a precision better than that of IRMAs. In conclusion, the results of this study indicate that CA 15-3 and CA 125 are satisfactorily assayed whereas CA 19-9 assay appears affected by larger differences between methods and by poorer precision of laboratories and kits.

Central nervous system (CNS) involvement in patients with systemic lupus erythematosus (SLE) is often difficult to evaluate because of protean neuropsychiatric (NP) manifestations and lack of reliable diagnostic markers. In the reported study the role of single-photon emission tomography (SPET) with technetium-99m hexamethylpropylene amine oxime (HMPAO) in the evaluation of CNS involvement in SLE was assessed and the relations between SPET perfusion defects, EEG examination, magnetic resonance imaging (MRI) findings and clinical presentation were examined. Twenty SLE patients with different NP manifestations were studied. Multiple areas of hypoperfusion, especially in the territory of the middle cerebral artery, were demonstrated by SPET analysis in all 20 patients. The number of hypoperfused areas and the degree of were more marked in patients with multiple NP manifestations. MRI and EEG evaluations were positive for 14 of 18 and for 12 of 20 patients, respectively. In the patients with positive SPET and MRI, 87 MRI focal lesions and 63 hypoperfused areas were found, and for 51 of these 63 at least one MRI lesion was found in the same anatomical region. SPET examination of patients with a normal EEG showed fewer hypoperfused areas and a lower degree of asymmetry compared to patients with an abnormal EEG. SPET of patients with focal EEG abnormalities showed more hypoperfused areas (difference not statistically significant) and a higher AI than did SPET of the patients with diffuse EEG abnormalities. Seven of 11 anatomical regions with focal EEG abnormalities. Seven of 11 anatomical regions with focal EEG abnormalities had co-localized hypoperfused areas and in two of these seven no detectable MRI lesions were found. The analysis of SPET and NP manifestations showed that 12 of 20 patients had at least one positive correlation, always involving the areas with the highest AI. In total, 51/88 (58%) hypoperfused areas correlated with the MRI findings and 31/88 (35%) with NP manifestations; for seven of the latter no concurrent MRI lesions were detected in the same anatomical region. It is concluded that SPET study of brain perfusion is a sensitive method for the evaluation of CNS involvement in SLE; furthermore, it is able to reveal disease progression and the lesions most relevant at the time of evaluation, and can objectify those NP manifestations without detectable MRI abnormalities. Nevertheless, because of the sensitivity of MRI in detecting morphological lesions, a complete evaluation of CNS involvement should be performed, combining SPET with MRI.

The aim of the study was to investigate the biologic significance of p21 expression in normal and neoplastic ovarian tissues.Western blotting analysis of p21/ras oncoprotein was conducted in a group of 14 normal and cystic ovaries, six benign tumors, 42 primary ovarian cancers, and 15 omental metastases.Levels of p21 were similar in normal and cystic ovaries and in benign tumors, whereas they were significantly higher in malignant tumors than in control tissues (median 1.91, range 0.12 to 5.00 vs median 1.03, range 0.32 to 2.20; p = 0.023) and in omental metastases than in primary ovarian carcinomas (median 3.05, range 0.55 to 5.72 vs median 1.97, range 0.12 to 5.00; p = 0.14). We found no correlation between p21 expression and histopathologic or clinical characteristics. Estrogen receptor-positive and progesterone receptor-positive tumors expressed higher p21 levels than did estrogen receptor-negative and progesterone receptor-negative tumors (p0.05), but no correlation with epidermal growth factor receptor status was found. In the univariate analysis of survival p21 positivity showed a negative prognostic value.The enhancement of p21 protein is associated in the ovarian tissue with the malignant phenotype and the acquisition of metastatic potential.

Objectives The measurement of plasma renin activity (PRA) is very convenient for estimating the action of the renin system, but its interlaboratory reproducibility is notoriously poor. This multicentre study aimed to examine whether an immunoradiometric assay which quantifies renin directly with monoclonal antibodies can reduce this limitation of the enzymatic assay. The study also aimed to establish the reference values of immunoreactive renin (IrR) in a large sample of normotensive subjects and patients with various pathophysiological conditions. Design and methods PRA and IrR were measured once in each of the eight participant centres in eight pool plasma samples with a wide range of renin content; in seven centres these measurements were repeated twice more in order to compare the intralaboratory interassay reproducibility of both methods. Finally, PRA and IrR were measured in the supine and standing positions in 503 subjects including normal controls, patients with various forms of hypertension, patients with Cushing's and Bartter's syndromes, patients with hepatic cirrhosis and pregnant women. Results We found that both the inter- and intralaboratory coefficients of variation for PRA measurements were higher than those for IrR. In plasma samples from normal subjects and from patients, mean +/- SEM supine PRA and IrR ranged, respectively, from 0.08 +/- 0.03 ng/ml per h and 2.6 +/- 0.5 pg/ml in patients with Conn's syndrome to 7.2 +/- 2.5 ng/ml per h and 138 +/- 51 pg/ml in patients with hepatic cirrhosis. PRA and IrR were found to be significantly correlated in all laboratories (mean +/- SEM of correlation coefficients 0.84 +/- 0.03) and for all of the conditions (correlation coefficient ranging from 0.98 in patients with Cushing's syndrome to 0.50 in pregnant women). However, for the pregnant women the slope of the regression line depicting the PRA-IrR relationship was significantly steeper than for all of the other conditions. Conclusions In our experience the inter- and intralaboratory reproducibilities of the immunoradiometric assay appear to be greater than can be achieved with the enzymatic assay, the difference being probably due to the greater complexity of the latter. The two methods provide superimposable information on the renin-angiotensin system activity, except in pregnancy, during which the PRA:IrR ratio is much higher than in the other conditions. Therefore, in this and other pathophysiological situations associated with marked angiotensinogen concentration alterations, the enzymatic assay may be still preferable for assessing the activity of the system accurately.

Objective. —To assess prospectively the impact on survival and health-related quality of life of two follow-up protocols in patients with early breast cancer. Design. —Randomized controlled clinical trial. Setting. —Multicenter study involving 26 general hospitals in Italy. Patients. —A consecutive sample of 1320 women younger than 70 years with stage I, II, and III unilateral primary breast cancer. Intervention. —Patients were randomly assigned to an intensive surveillance, which included physician visits and performance of bone scan, liver echography, chest roentgenography, and laboratory tests at predefined intervals (n=655), or to a control regimen (n=665), in which patients were seen by their physicians at the same frequency but only clinically indicated tests were performed. Both groups received a yearly mammogram aimed at detecting contralateral breast cancer. Main Outcome Measures. —Primary end points were overall survival and health-related quality of life. Results. —Compliance to the two follow-up protocols was more than 80%. At a median follow-up of 71 months, no difference was apparent in overall survival with 132 deaths (20%) in the intensive group and 122 deaths (18%) in the control group. No significant differences were apparent in time to detection of recurrence between the two groups. Measurements of health-related quality of life (ie, overall health and quality-of-life perception, emotional well-being, body image, social functioning, symptoms, and satisfaction with care) at 6,12, 24, and 60 months of follow-up did not show differences by type of care received. Conclusions. —Results of this trial support the view that a protocol of frequent laboratory tests and roentgenography after primary treatment for breast cancer does not improve survival or influence health-related quality of life. Routine use of these tests should be discouraged. (JAMA. 1994;271:1587-1592)

This article illustrates the two main methods for routine measurement of cytoplasmic estrogen receptor status in neoplastic biopsy. The first is the Dextran Coated Charcoal Technique (D.C.C. Assay) which is still the method of choice in the majority of clinical laboratories for its simplicity, reproducibility and low cost. The second is a more advanced technique based on the specific binding, enzimatically displayed, of commercially available antiestrogen monoclonal antibodies (Enzyme Immuno Assay - ABBOTT).The sui generis characteristics of endocrine sensitivity assessment on tumor tissues and the importance of decision-making connected with the assay justify rigorous quality assurance schemes. The quality control design proposed by the Italian Committee concerned the evaluation of several lyophilized preparations with scalar receptor content; this permits the identification through linear regression analysis of systematic and non-systematic errors. The Italian Committee has currently connected 50 labs from most regions of the country.

The possible relationships between hormone receptor status and several clinical (age, gynecologic history, clinical stage) and morphologic aspects (histologic grade, vascular invasion, lymphocytic infiltration, necrosis, fibrosis, elastosis and lymph node metastasis) were evaluated. A highly significant correlation between estrogen receptor levels, patient age, menses regularity and postmenopausal status was found. The histologic features most significantly related to tumor receptor status were histologic grade, lymphocytic infiltrate, necrosis and elastosis. Since these same histologic aspects appear to influence prognosis in breast cancer, the prognostic significance attributed to tumor receptor levels is substantiated. Therefore the importance of this assay is confirmed, not only for its diagnostic and therapeutic purposes, but also for its prognostic value.