The purpose of this study was to determine the safety, distribution, internal dosimetry, and initial human epidermal growth factor receptor 2 (HER2)-positive tumor images of 64 Cu-DOTA-trastuzumab in humans. Methods: PET was performed on 6 patients with primary or metastatic HER2-positive breast cancer at 1, 24, and 48 h after injection of approximately 130 MBq of the probe 64 Cu-DOTA-trastuzumab. Radioactivity data were collected from the blood, urine, and normal-tissue samples of these 6 patients, and the multiorgan biodistribution and internal dosimetry of the probe were evaluated. Safety data were collected for all the patients after the administration of 64 Cu-DOTA-trastuzumab and during the 1-wk follow-up period. Results: According to our results, the best timing for the assessment of 64 Cu-DOTA-trastuzumab uptake by the tumor was 48 h after injection. Radiation exposure during 64 Cu-DOTA-trastuzumab PET was equivalent to that during conventional 18 F-FDG PET. The radioactivity in the blood was high, but uptake of 64 Cu-DOTA-trastuzumab in normal tissues was low. In 2 patients, 64 Cu-DOTA-trastuzumab PET showed brain metastases, indicative of blood-brain barrier disruptions. In 3 patients, 64 Cu-DOTA-trastuzumab PET imaging also revealed primary breast tumors at the lesion sites initially identified by CT. Conclusion: The findings of this study indicated that 64 Cu-DOTA-trastuzumab PET is feasible for the identification of HER2-positive lesions in patients with primary and metastatic breast cancer. The dosimetry and pharmacologic safety results were acceptable at the dose required for adequate PET imaging.
Background: There are several clinical diagnostic criteria for allergic bronchopulmonary aspergillosis (ABPA). However, these criteria have not been validated in detail, and no criteria for allergic bronchopulmonary mycosis (ABPM) are currently available. Objective: This study proposes new diagnostic criteria for ABPA/ABPM, consisting of 10 components, and compares its sensitivity and specificity to existing methods. Methods: Rosenberg-Patterson criteria proposed in 1977, the International Society for Human and Animal Mycology (ISHAM) criteria proposed in 2013, and this new criteria were applied to 79 cases with pathological ABPM and the control population with allergic mucin in the absence of fungal hyphae (n 5 37), chronic eosinophilic pneumonia (n 5 64), Aspergillussensitized severe asthma (n 5 26), or chronic pulmonary aspergillosis (n 5 24). These criteria were also applied to the 179 cases with physician-diagnosed ABPA/ABPM in a nationwide Japanese survey. Results: The sensitivity for pathological ABPM with Rosenberg-Patterson criteria, ISHAM criteria, and this new criteria were 25.3%, 77.2%, and 96.2%, respectively. The sensitivity for physician-diagnosed ABPA/ABPM were 49.2%, 82.7%, and 94.4%, respectively. The areas under the curve for the receiveroperating characteristic curves were 0.85, 0.90, and 0.98, respectively. The sensitivity for ABPM cases that were culture-positive for non-Aspergillus fungi were 13.0%, 47.8%, and 91.3%, respectively. Conclusions: The new diagnostic criteria, compared with existing criteria, showed better sensitivity and specificity for diagnosing ABPA/ABPM. (J Allergy Clin Immunol 2020;nnn:nnn-nnn.)
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The extracellular poly(3-hydroxybutyrate) depolymerase gene from Akaligenes faecalis Tl was cloned into Escherichia coli DH1 by using the plasmid pUC8. An A. faecalis Ti genomic library was prepared in E. coli from a partial Sau3AI digest and screened with antibody against the depolymerase. Of the 29 antibody-positive clones, 1 (pDP14), containing about 4 kilobase pairs of A. (22), modified by the addition of achromopeptidase (TBL-1; 1.5 mg/ml) (12) to the lysozyme-EDTA solution to lyse the organism. Plasmid DNA was isolated from a chloramphenicol-amplified culture (18) of E. coli by the method of Bimboim and Doly (1) and then purified by gel filtration. A. faecalis Ti DNA was partially digested with Sau3AI, and fragments of 4 to 9 kilobase pairs (kbp) in size were isolated from the agarose gel by the glass powder method (28). The size-fractionated DNAs were ligated into BamHI-digested and alkaline phosphatase-treated pUC8, using T4 ligase. E. coli DH1 was transformed with recombinant plasmid DNA by the calcium chloride method of Mandel and Higa (17), and ampicillinresistant (Apr) transformants were selected and immunologically screened (9) with anti-PHB depolymerase immunoglobulin G raised in a rabbit.Restriction mapping and subcloning. Mapping of restriction sites was performed by the standard procedure (18). Deletion derivatives were constructed by digesting plasmids with a single restriction enzyme, followed by ligation of the products. Subcloning was performed by isolating DNA fragments from 0.8% agarose gels by electroelution onto DEAE-paper (6). These fragments were then ligated into pUC8 that had been cut with appropriate restriction enzymes. pUC8 with subcloned DNA fragments was introduced into the E. coli JM103 recipient by transformation, and white colonies containing recombinant plasmids were 184 JOURNAL
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