The concept of minimum detectable activity of radionuclide activity meters and their suitability for routine quality control of radiopharmaceuticals. An experimental study.

Radionuclide activity meters ("dose calibrators") are ionization chambers designed to measure relatively high amount of activities which are normally contained in radiopharmaceuticals. However, in the current radiopharmacy practice, these radiation detectors have been proposed to be used in measurements of samples with lower activity, such as in routine quality control (QC) tests. To check the feasibility of such measurements, in this work we assessed the performance of four different devices in the lower range of detectability, by means of experimental measurements of a radioactive sample. Accuracy and precision of each device was evaluated as a function of the activity contained in the sample in order to estimate a threshold value, or minimum detectable activity (MDA), which, according to our operational definition, may be used to express the concept of Limit of Quantification (LoQ). Moreover, a generalized procedure for the estimation of the MDA was established, which, being device- and radionuclide-independent, it may be adopted by every laboratory. Our results showed a significant variability in the MDA achieved by different activity meters. Hence a single QC test may result feasible with one specific instrument, and not with another one. Moreover, feasibility depends also on the confidence level required for each test. For these reasons, each activity meter should be qualified for its MDA or LoQ by each laboratory according to a procedure such as that described in this paper.

Use of 65Zn as a tracer for the assessment of purification in the 68Ga-DOTANOC synthesis.

In the last years (68)Ga has got into the focus of researchers and clinicians especially for radio-labeling of biomolecules; an important characteristic of this positron emitting isotope is its availability via the (68)Ge/(68)Ga generator system: the long-lived (68)Ge (t1/2=270.8 d) produces the short-lived (68)Ga (t1/2=67.63 min) which decays to stable (68)Zn. (68)Ge breakthrough compromises (68)Ga radionuclidic purity, while (68)Zn might affect the specific activity of the radiopharmaceutical. In this paper we investigated the weight of these impurities in (68)Ga-DOTANOC synthesis. (65)Zn (t1/2=244.26d; decay mode: EC 98.3%, ß(+) 1.7%) was used as a radiotracer of stable (68)Zn; samples of the purification columns, wastes and product were recovered and measured with a calibrated HPGe gamma-ray spectrometry system. The results showed that (68)Zn competes with (68)Ga in labeling DOTANOC with a (95±2)% labeling yield; they also proved the effectiveness of the STRATA X-C cationic post-processing of the generator eluate in lowering the amount of this impurity to less than 1%. Moreover this approach, along with the purification of the final product through a STRATA X cartridge, effectively removes (68)Ge breakthrough providing a (68)Ga-DOTANOC radionuclidic purity of (99.9999986±0.0000006)%, superior to 99.9% required by the Pharmacopoeia Monograph on (68)Ga Edotreotide injection.

Effective production of 65Zn with a PET cyclotron.

Cyclotrons dedicated to the production of PET radiopharmaceuticals for clinical use can be a resource also for research in other fields of science. In the present study we assessed the feasibility of (65)Zn production via the (65)Cu(p,n)(65)Zn reaction using a 16.5 MeV GE-PETtrace biomedical cyclotron. The radioisotope (65)Zn (t(1/2)=244.26 zd; decay mode: EC 98.3%, ß(+) 1.7%) has a wide range of applications, especially in botany and agriculture; its long half life allows storage and delivery to research centers without a cyclotron on site. Natural copper foils (thickness: 100 µm; purity: >99.9%) were irradiated in a solid target station developed in our institution. The EXFOR and IAEA cross sections databases were carefully studied to optimize the irradiation setup in order to maximize the yield of the radionuclide of interest and minimize the production of isotopic impurities. We performed production runs at 20 µA for 30 min. Activity was measured using a calibrated HPGe gamma ray detector and an ionization chamber radionuclide activity meter. These measures showed the presence of two main contaminants: (63)Zn (t(1/2)=38.47 min) and (64)Cu (t(1/2)=12.7h); we produced (65)Zn with a saturation yield of (1.06 ± 0.07) GBq/µA (E(p)=12.65→10.48 MeV). The irradiated target was dissolved in 1 ml of 6N HNO(3) at the temperature of 90°C and then purified through anion exchange chromatography with BIO RAD AG 1×8 resin. The radionuclidic purity of the final solution of (65)Zn in HCl 0.005 N, expressed in activity, resulted superior to (99.921 ± 0.003) %.

Prediction of (89)Zr production using the Monte Carlo code FLUKA.

The widely used Monte Carlo simulation code FLUKA has been utilized to prototype a solid target for the production of (89)Zr by irradiation of a metallic (89)Y target foil in a 16.5MeV proton biomedical cyclotron, through the reaction (89)Y(p, n)(89)Zr. Simulations were performed with and without an Al energy degrader. In the setup of the geometry of the target, state of the art support tools, like SimpleGeo, were used for accurate, detailed modeling. The results permitted a quick assessment of all possible radionuclidic contaminants and confirmed that the use of an energy degrader avoids production of the most important impurity, (88)Zr. The estimated value for the activity produced in one hour of irradiation at 20µA is 384 ± 42MBq; this is encouraging, indicating possible production of clinically significant amounts of activity with the relatively simple target setup adopted. Initial experimental tests gave results in excellent agreement with simulations, confirming the usefulness and accuracy of FLUKA as a tool for the design and optimization of targets for the production of PET radionuclides.

Experimental monitoring of ozone production in a PET cyclotron facility.

Ozone produced from radiolytic processes was investigated as a possible health hazard in the working environment at the University Hospital "S.Orsola--Malpighi" PET facility. Intense radiation fields can generate ozone, known to be the most toxic gas produced by ionizing radiation around a particle accelerator. To evaluate ozone concentration in air, two different measurement campaigns were conducted with passive diffusion detectors. Comparison of the results with the concentration limits recommended by American Conference of Governmental Industrial Hygienists (ACGIH) demonstrated that ozone poses no health hazard to workers around a biomedical cyclotron.

First Report of Fusarium andiyazi Associated with Rice Bakanae in Italy.

Rice (Oryza sativa L.) is cultivated on approximately 230,000 ha in northern Italy. Since 2001, increasing economical losses presumably caused by Fusarium fujikuroi Nirenberg (Gibberella fujikuroi mating population C), an exotic fungus known as the etiological agent of Bakanae disease, have been reported in Italy. The spread of this disease is primarily seedborne. In 2009, during an annual survey of Italian rice seed, 69 samples were tested for the presence of strains belonging to the G. fujikuroi species complex. Four hundred seeds per sample were surface sterilized and then placed in 90-mm Petri dishes containing potato dextrose agar and incubated for 7 days at 21°C. Thirty two putative G. fujikuroi strains were single-spore purified and identified on the basis of their morphological features on Spezieller Nährstoffarmer agar plates with a piece of sterile filter paper. Strains were characterized at species level by morphological observations (1,2) and translation elongation factor 1-α (TEF) gene sequencing. Unexpectedly, 60% of the strains evaluated belonged to the species F. andiyazi Marasas, Rheeder, Lampr., K.A. Zeller & J.F. Leslie. This fungus, first described on sorghum (Sorghum bicolor L.) in Africa and the United States (1), has been reported to be one of the species associated with Bakanae in Asia and Africa (3). Two F. andiyazi strains, (E432 and E439), isolated in the district of Modena were chosen for pathogenicity testing and their TEF gene sequences were deposited in GenBank (Accession Nos. GU827420 and GU827419). A conidial suspension was produced on Mung-bean liquid media and adjusted to a concentration of 1 × 106 CFU/ml. Italian cv. Galileo was used in the test because of its high susceptibility to Bakanae (Ente Nazionale delle Sementi Elette, Verona, Italy, data unpublished). Rice seeds were heat sterilized for 20 min at 60°C, submerged for 30 min in the conidial suspensions, dried, and subjected to a blotter test. Uninoculated, sterilized seeds served as a control. Seeds were incubated for 15 days in a growth chamber (26°C, 80% relative humidity, and 12-h photoperiod). For each strain, the experiment was repeated three times on samples of 25 seedlings. Results were analyzed by analysis of variance and Tukey test. Symptoms consisted of a generic seedling wilt, a root length reduction ranging from 21 to 48%, and the presence of root discoloration. Seed germination was reduced by 9%. Shoot development was not significantly altered. Proof of pathogenicity was obtained through reisolation of F. andiyazi from symptomatic tissues. To our knowledge, this is the first report of F. andiyazi on rice in Europe. References: (1) W. F. O. Marasas et al. Mycologia 93:1203, 2001. (2) H. I. Niremberg and K. O'Donnell. Mycologia 90:434, 1998. (3) E. G. Wulff et al. Environ. Microbiol. 12:649, 2009.

Radiolabelling, quality control and radiochemical purity assessment of the Octreotide analogue 68Ga DOTA NOC.

Somatostatin receptors 1-5 are over expressed in neuroendocrine tumours (NETs). 68Ga-labelled [1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid]-1-Nal3-Octreotide (DOTA NOC), a recent synthesized somatostatin analogue, shows high affinity for those receptors. Herein, modifications of a commercial module for the labelling of DOTA NOC with 68Ga, as well as the assessment of time course of the radiochemical purity variation are described. The evaluation of radiochemical stability was done by two different chromatographic methods: reversed-phase radio HPLC and fast TLC analysis. Labelled compound has been found radiochemically stable within 3h from the end of labelling (EOL) and radiochemical purity was always higher than 99%. After 73 labelling sessions the system showed great reproducibility and high radiochemical yield.

Quality control of 99Mo/99Tcm generators: results of a survey of the Radiopharmacy Working Group of the Italian Association of Nuclear Medicine (AIMN).

A multicentre survey of the quality control of 99Tcm 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 99Mo content was 3.8 x 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.

Evaluation of lung clearance of inhaled pertechnegas.

Pertechnegas is a new ventilation agent produced by modifying the atmosphere of combustion of Technegas. Due to its rapid disappearance from the lungs, Pertechnegas has been suggested as useful in measuring pulmonary epithelial permeability. This study aimed to assess the reliability of ventilation scans with Pertechnegas to evaluate alveolar-capillary permeability. Six non-smokers with no evidence of pulmonary disease were investigated. Scintigraphic data were used to evaluate the site of Pertechnegas deposition (by assessing the Penetration Index [PI] of the gas), its clearance rate (by calculating the time to half-clearance [T1/2]) and its lung distribution (by means of a pixel-by-pixel analysis. PI measurements produced a mean value of 88.8 +/- 13.3% (range 69-117%). Time activity curves showed a fast clearance in all cases (mean T1/2 = 10.7 +/- 2.1 min, range 8.1-14.3 min). Comparison of statistical indices of uniform deposition (skewness and kurtosis) indicated satisfactory homogeneity of Pertechnegas distribution throughout the lungs. These data show that after inhalation Pertechnegas has a peripheral deposition and a homogeneous distribution in the lungs and is rapidly cleared through the alveolar-capillary barrier. In conclusion Pertechnegas can be recommended as a potential radiopharmaceutical for studying the pulmonary epithelial barrier.

[Amides of amino acids and peptides as antifungal agents]. / Ammidi di amminoacidi e peptidi come antifungini.

The synthesis of pyrazolyl-amides of aminoacids and peptides is described. The chemicals were tested for antifungal activity against wheat powdery mildew (Erysiphe graminis DC.), cucumber powdery mildew (Erysiphe cichoracearum DC.), wheat brown rust (Puccinia recondita Rob. ex Desm. f. sp. tritici Erikss et Henn.), celery leaf spot (Septoria Apii Briosi ed Cav. Chest.) and collar rot (Rhizoctonia solani Kuhn). Some of these compounds showed antifungal activity.

[Derivatives of 4-nitroso-aminopyrazole as antifungal agents]. / Derivati del 4-nitroso-5-amminopirazolo come antifungini.

The synthesis of 4-nitroso-5-amminopyrazoles and of 4-nitroso-5-pyrazolylurethans and -ureas is described. The chemicals were tested for antifungal activity against Erysiphe graminis, Erysiphe cichoracearum, Puccinia recondita, Septoria apii and Rhizoctonia solani. A number of the described compounds showed some antifungal activity.