Efficacy of <Superscript>99m</Superscript>Tc pertechnetate and <Superscript>131</Superscript>I radioisotope therapy in sodium/iodide symporter (NIS)-expressing neuroendocrine tumors in vivo

Purpose There is growing interest in the human sodium/iodide symporter (NIS) gene both as a molecular imaging reporter gene and as a therapeutic gene. Here, we show the feasibility of radioisotope therapy of neuroendocrine tumors. As a separate application of NIS gene transfer, we image NIS-expressing tumors with pinhole SPECT in living subjects. Methods Biodistribution studies and in vivo therapy experiments were performed in nude mice carrying stably NIS-expressing neuroendocrine tumor xenografts following i.v. injection of ¹³¹I and ^99mTc pertechnetate. To show the usefulness of NIS as an imaging reporter gene, ^99mTc pertechnetate uptake was imaged in vivo using a clinical gamma camera in combination with a custom-made single pinhole collimator, followed by SPECT/small animal MRI data coregistration. Results NIS-expressing neuroendocrine tumors strongly accumulated ¹³¹I and ^99mTc pertechnetate, as did thyroid, stomach, and salivary gland. The volume of NIS-expressing neuroendocrine tumors decreased significantly after therapeutic administration of ¹³¹I or ^99mTc pertechnetate, whereas control tumors continued to grow. NIS-mediated uptake of ^99mTc pertechnetate could be imaged in vivo at high resolution with a clinical gamma camera equipped with a custom-made single pinhole collimator. High-resolution functional and morphologic information could be combined in a single three-dimensional data set by coregistration of SPECT and small animal MRI data. Lastly, we demonstrated a therapeutic effect of ^99mTc pertechnetate on NIS-expressing neuroendocrine tumors in cell culture and, for the first time, in vivo, thought to be due to emitted Auger and conversion electrons. Conclusions NIS-expressing neuroendocrine tumors efficiently concentrate radioisotopes, allowing for in vivo high-resolution small animal SPECT imaging as well as rendering possible successful radioisotope therapy of neuroendocrine tumors.     

Executive functions in borderline personality disorder

Different domains of executive function such as working memory and response inhibition were investigated together with elementary cognitive processes in borderline personality disorder (BPD). Patients with BPD (N=28) were compared to nonpatient controls (NP, N=28) on eight tasks (e.g. n-back, Go/NoGo, CPT-AX). In order to separate impairments in different cognitive domains and to assess the influence of more elementary cognitive processes on executive functioning, tasks were embedded in a reaction-time-decomposition approach. BPD patients solved tasks with accuracies comparable to those of nonpatients. The only exception was the n-back task, for which working memory is required: here, error rates were higher and increased more prominently in BPD patients depending on working memory load. In most tasks, movement times were shorter for BPD patients than for nonpatients, while the quality of task-solving was comparable. The faster processing in the BPD group was observable starting with the simplest task, i.e. a simple reaction-time task. These findings suggest that domains of executive functioning are differentially affected in BPD. In contrast to load-dependent deficits in working memory, response inhibition processes were unimpaired. Faster action-related processes could be observed in BPD patients in a variety of tasks; however, these did not influence executive functioning.     

Adenocarcinoma of the cervix

BACKGROUND: The current study examines 1) the sensitivity of detection of invasive adenocarcinoma of the cervix in a routine cervical screening service, and 2) the frequency in smears of cytologic criteria previously found to be useful in diagnosis. METHODS: Data on women with diagnoses of adenocarcinoma of the cervix accessioned at the Western Australian Cervical Cytology Registry during the period 1993-1998 were examined, where smears had been reported by Western Diagnostic Pathology within three years of the biopsy diagnosis. Smears and biopsy material were reviewed. RESULTS: Thirty-six smears from 24 women were reviewed. Of those, 58.3% had been reported as a possible or definite high grade epithelial abnormality (HGEA). On review it was thought that this could be improved to 77.8%. The screening or diagnostic error was thus 19.4% and the sampling error 22.2%. The likelihood of an individual woman receiving a report of a possible or definite HGEA in the three years before biopsy was 83.3%. In retrospect this could have been improved to 91.7%. Heavy bloodstaining with abundant abnormal glandular epithelium (14 smears) and small three-dimensional or papillary clusters (16 smears) were the most frequent clues to invasion. Tumor necrosis/diathesis was present in eight smears, but easily seen in only four, while marked nuclear pleomorphism and macronucleoli were seen in three and one smears respectively. In cases with a discrepancy between the initial and the review findings, very small amounts of abnormal material (three smears), a resemblance to endometrial cells (one smear), and an unusual appearance of folded monolayered sheets (three smears) contributed to the difficulty of diagnosis. CONCLUSIONS: There were significant sampling and screening/diagnostic errors (22.2% and 19.4%, respectively). Screening and diagnostic errors could perhaps be reduced by a greater awareness of the range of cytologic changes, but these may be subtle. Heavy bloodstaining with abundant abnormal glandular material may be a useful clue to invasive, rather than in situ, adenocarcinoma, even in the absence of tumor diathesis or fully malignant nuclear criteria.     

Identification of urinary protein pattern in type 1 diabetic adolescents with early diabetic nephropathy by a novel combined proteome analysis

Diabetic nephropathy is the main cause of morbidity and mortality in patients with Type 1 diabetes mellitus. Microalbuminuria has been established as a risk factor for the development and the progression of diabetic renal disease. A strong demand exists for better technologies to provide accurate diabetic nephropathy risk estimates before renal functional or structural disturbances already become established. Here, we present the application of a novel proteomics technology identifying urinary polypeptides and proteins. In this pilot study, we investigated 44 Type 1 diabetic patients with more than 5 years of diabetes duration compared with an age-matched control group. Random spot urine samples were examined utilizing high-resolution capillary electrophoresis (CE), coupled to mass spectrometry (MS). More than 1000 different polypeptides, characterized by their separation time and mass, were found between 800 Da and 66.5 kDa. Mathematical analysis revealed specific clusters of 54 polypeptides only found in Type 1 diabetic patients and an additional 88 polypeptides present or absent in patients with beginning nephropathy defined by the albumin-to-creatinine ratio (ACR; >35 mg/mmol). We observed polypeptide patterns characteristic for healthy controls and diabetic patients and subdivision of patients according to the excretion of polypeptides typical for diabetic nephropathy. Our study revealed that the urinary proteome contains a much greater variety of polypeptides than previously recognized and demonstrated the successful application of a novel high-throughput technology towards the human urinary proteome. Future prospective studies with the application of this technique may enable the earlier and more accurate detection of individuals at high risk to develop diabetic nephropathy.     

Chemoembolization of rat liver metastasis with microspheres and gemcitabine followed by evaluation of tumor cell load by chemiluminescence

An orthotopic, isogenic rat model was used to determine the potential of chemoembolization (CHE) for reducing the tumor cell load of a diffusely metastatic liver. Seven days after injecting CC531-lac-Z cells intraportally to male WAG/Rij rats, tumor positive animals were treated by injection into the hepatic artery with solvent (n=17), degradable starch microspheres (DSM, 30 mg/kg; n=16), DSM plus 5-fluorouracil (5-FU, dosages: 90, 60, and 40 mg/kg) or DSM plus gemcitabine (Gem, dosages: 100, 80, 50, and 10 mg/kg). After 3 more weeks the experiment was terminated, the livers were weighted and the number of CC531-lac-Z cells per liver was determined. Injection of DSM reduced the tumor cell load by 21% (T/C%=79), the combination with 5-FU caused a stimulation of growth at 40 mg/kg (T/C%=291; n=10), but effected dose-dependent reductions in tumor cell number at 60 mg/kg (T/C%=86; n=16), and 90 mg/kg (T/C=19; n=17). None of these effects was significantly different from controls. The combination of DSM plus Gem was toxic at the highest dose (100 mg/kg), but well tolerated and highly effective at 80 mg/kg (T/C%= 16; n=12), 50 mg/kg (T/C%=9; n=12), and 10 mg/kg (T/C%=26; n=14). These results were significantly different from controls (p<0.05), respectively. In summary, the comparison of CHE with 5-FU or Gem shows that the efficacy of Gem in reducing the hepatic tumor cell load was significantly higher and its therapeutic ratio was greater than that of 5-FU.     

Unilateral choanal atresia: surgical technique and long-term results

Although transnasal techniques to treat choanal atresia have become the standard, risks remain of restenosis of the opened choana requiring dilatations. In addition, a new technique without stenting and requiring less invasive post-operative care would be an improvement. With this in mind, in a non-randomized trial, a modification of the transnasal surgical repair was used in eight patients with unilateral choanal atresia by which no stents were used. While the standard procedure consists of partial resection of the bony septum and the atretic plate, we additionally removed lower parts of the anterior wall and the floor of the sphenoid sinus, creating a new airway canal from the nasal cavity to the sphenoid into the epipharynx. Post-operative care by the patient and the surgeon was noted. The success of choanal patency was followed by nasal endoscopy and rhinomanometry with a mean post-operative follow-up time of 1.9 years. In all patients, both choanae remained patent confirmed by nasal endoscopy and rhinomanometry showing almost symmetric resistance. Post-operative care comprised antibiotic prophylaxis and nasal douching with saline solution by the patient; no further treatment by the surgeon was necessary. According to these long-term results we would recommend this type of surgery for two reasons: the use of stents no longer necessary and post-operative care by the surgeon can be minimized.     

Expression of the B7-related molecule B7-H1 by glioma cells: a potential mechanism of immune paralysis

Human glioblastoma is a highly lethal tumor that is known for its immune inhibitory capabilities. B7-homologue 1 (B7-H1), a recently identified homologue of B7.1/2 (CD80/86), has been described to exert costimulatory and immune regulatory functions. We investigated the expression and the functional activity of B7-H1 in human glioma cells in vitro and in vivo. Although lacking B7.1/2 (CD80/86), all 12 glioma cell lines constitutively expressed B7-H1 mRNA and protein. Exposure to IFN-gamma strongly enhanced B7-H1 expression. Immunohistochemical analysis of malignant glioma specimens revealed strong B7-H1 expression in all 10 samples examined, whereas no B7-H1 expression could be detected on normal brain tissues. To elucidate the functional significance of glioma cell-related B7-H1 expression, we performed coculture experiments of glioma cells with alloreactive CD4+ and CD8+ T cells. Glioma-related B7-H1 was identified as a strong inhibitor of CD4+ as well as CD8+ T-cell activation as assessed by increased cytokine production (IFN-gamma, interleukin-2, and interleukin-10) and expression levels of the T-cell activation marker (CD69) in the presence of a neutralizing antibody against B7-H1 (mAb 5H1). B7-H1 expression may thus significantly influence the outcome of T-cell tumor cell interactions and represents a novel mechanism by which glioma cells evade immune recognition and destruction.     

Phylogenetic relationships and new records of Asterinaceae (Dothideomycetes) from Panama

Asterinaceae are small obligately biotrophic pathogens growing superficially on living leaves of higher plants in tropical and subtropical regions worldwide. The species-rich but rarely studied ascomycete group has an uncertain placement within the Dothideomycetes because molecular data are missing completely up to now. Based on nuclear DNA from fresh material of five Asterina spp. and a related anamorphic stage from Panama, we present the first molecular phylogenetic hypothesis of the Asterinaceae within the Dothideomycetes. A combined SSU and LSU rDNA phylogenetic analysis shows that species of Asterina and its anamorphs form a well supported monophyletic clade within the Dothideomycetes with Venturiaceae as sistergroup. Three Asterina spp. included in the molecular study are new records for Panama and Central America with new records of host plant species: A. cestricola on Cestrum rugulosum (Solanaceae), A. weinmanniae on Weinmannia pinnata (Cunoniaceae) and A. zanthoxyli on Zanthoxylum scheryi (Rutaceae). A. cestricola and A. weinmanniae are illustrated here for the first time.