Toward a clinical application of ex situ boron neutron capture therapy for lung tumors at the RA-3 reactor in Argentina.

PURPOSE: Many types of lung tumors have a very poor prognosis due to their spread in the whole organ volume. The fact that boron neutron capture therapy (BNCT) would allow for selective targeting of all the nodules regardless of their position, prompted a preclinical feasibility study of ex situ BNCT at the thermal neutron facility of RA-3 reactor in the province of Buenos Aires, Argentina. (l)-4p-dihydroxy-borylphenylalanine fructose complex (BPA-F) biodistribution studies in an adult sheep model and computational dosimetry for a human explanted lung were performed to evaluate the feasibility and the therapeutic potential of ex situ BNCT. METHODS: Two kinds of boron biodistribution studies were carried out in the healthy sheep: a set of pharmacokinetic studies without lung excision, and a set that consisted of evaluation of boron concentration in the explanted and perfused lung. In order to assess the feasibility of the clinical application of ex situ BNCT at RA-3, a case of multiple lung metastases was analyzed. A detailed computational representation of the geometry of the lung was built based on a real collapsed human lung. Dosimetric calculations and dose limiting considerations were based on the experimental results from the adult sheep, and on the most suitable information published in the literature. In addition, a workable treatment plan was considered to assess the clinical application in a realistic scenario. RESULTS: Concentration-time profiles for the normal sheep showed that the boron kinetics in blood, lung, and skin would adequately represent the boron behavior and absolute uptake expected in human tissues. Results strongly suggest that the distribution of the boron compound is spatially homogeneous in the lung. A constant lung-to-blood ratio of 1.3 ± 0.1 was observed from 80 min after the end of BPA-F infusion. The fact that this ratio remains constant during time would allow the blood boron concentration to be used as a surrogate and indirect quantification of the estimated value in the explanted healthy lung. The proposed preclinical animal model allowed for the study of the explanted lung. As expected, the boron concentration values fell as a result of the application of the preservation protocol required to preserve the lung function. The distribution of the boron concentration retention factor was obtained for healthy lung, with a mean value of 0.46 ± 0.14 consistent with that reported for metastatic colon carcinoma model in rat perfused lung. Considering the human lung model and suitable tumor control probability for lung cancer, a promising average fraction of controlled lesions higher than 85% was obtained even for a low tumor-to-normal boron concentration ratio of 2. CONCLUSIONS: This work reports for the first time data supporting the validity of the ovine model as an adequate human surrogate in terms of boron kinetics and uptake in clinically relevant tissues. Collectively, the results and analysis presented would strongly suggest that ex situ whole lung BNCT irradiation is a feasible and highly promising technique that could greatly contribute to the treatment of metastatic lung disease in those patients without extrapulmonary spread, increasing not only the expected overall survival but also the resulting quality of life.

Qualitative autoradiography with polycarbonate foils enables histological and track analyses on the same section.

Neutron autoradiography is an imaging methodology that enables analysis of the spatial distribution of heavy ion emitters in a given material. In particular, it allows localization of (10)B in a tissue section put in contact with a nuclear track detector. Boron imaging is essential when considering boron neutron capture therapy as an option for treating cancerous tumors. A description of the autoradiography method is presented together with specific characteristics and technical details developed in our laboratory. We propose a new mounting technique to compare autoradiography images with the same section that gave rise to the latent tracks. The solid state nuclear track detector is polycarbonate, because it can be processed rapidly to obtain the autoradiographic results. It is a transparent material, which allows visualization of the sections mounted on it. Tissue can be removed easily and background is minimal.

10B concentration evaluation in autoradiography images by optical density measurements.

The quantification and analysis of the tracks forming the autoradiography image of a tissue section is essential for the measurement of particle emitter concentration and distribution (e.g. (10)B) in the sample. Conventional counting techniques cannot be used when track density is high because of track overlapping. A densitometry supported by image analysis method suitable for these cases has been developed. Optical density measurements obtained for boron solutions of known concentrations showed a linear behavior in the range of concentrations under consideration.

Reference systems for the determination of 10B through autoradiography images: application to a melanoma experimental model.

The amount of (10)B in tissue samples may be determined by measuring the track density in the autoradiography image produced on a nuclear track detector. Different systems were evaluated as reference standards to be used for a quantitative evaluation of boron concentration. The obtained calibration curves were applied to evaluate the concentration of (10)B in melanoma tumour of NIH nude mice after a biodistribution study. The histological features observed in the tissue sections were accurately reproduced by the autoradiography images.

Low energy proton irradiation effects on alveolar macrophages from young and aged rats.

A prospective study was undertaken to assess the radiotoxicity of accelerated particles in pulmonary alveolar macrophages (AM). We evaluated the effects of a single dose (10-75 Gy) of an external low-energy (20 MeV) proton beam on cultured AM oxidative metabolism and phagocytic function. Macrophages are the first line of defense against invading pathogens and are known to generate superoxide anion (O2), nitric oxide (NO), and mediators of antimicrobial and antitumoral defense mechanisms. We obtained AM by bronchoalveolar lavage from young (1-2 month old) and aged (9-12 month old) male Wistar rats. Cell viability, phagocytosis, O2 and NO production in control and proton-irradiated cultured AM were evaluated The effect of proton irradiation on cell viability was dose-dependent The higher doses induced a dramatic decrease in viability in the aged population. Phagocytosis increased 1.3-1.4 fold inboth populations irrespective of the dose delivered. Generation of O2 was always higher in the aged population for all the doses assayed and showed no significant variation from the control values. In the young population a clear increase was observed with doses of 25 and 50 Gy. NO production in AM from young animals rose in a dose-dependent manner. Conversely, proton irradiation did not affect NO production in macrophages from aged animals. The results of this study demonstrate that AM isolated from young and aged rats are functionally different and show a distinct behavior when exposed to proton irradiation. These findings suggest that age may condition response and must be taken into account when accelerated particle-radiotherapy protocols are considered as a valid therapeutic option for the treatment of cancer. To the best of our knowledge, this is the first report comparing sham-irradiated and proton-irradiated young and aged AM.

In vivo response to the different LET regions of a 55 MeV helium ion beam.

We evaluated the in vivo response to heavy particle irradiation in rat tail epidermis using silver-stained nucleolar organizer regions (AgNOR) as the end-point. The energy degradation of the beam across the circular section of the tail allowed us to study the damage elicited by two different LET regions of a helium beam, i.e. non-Bragg peak (NBP) and Bragg peak (BP), at different sites on the same sample. The tails were locally irradiated with a helium ion beam at different fluences. AgNOR exhibited marked variations between tissue areas only a few micrometers apart within each tail exposed to a given beam fluence. An analysis of the AgNOR variations in NBP and BP areas of tails exposed to different beam fluences revealed a dose-dependent effect. The AgNOR provide quantitative evidence of differential damage in neighboring tissue areas exposed to different LET regions of a helium-ion beam.

Humoral (antibody-mediated) rejection in lung transplantation.

To confirm the existence and characterize the pathologic features of humoral (antibody-mediated) lung rejection, we prospectively studied 55 lung transplant recipients (24 male [44%] and 31 female [56%], age range 14 to 69 years [mean 45]). The time between transplantation and biopsy ranged from 2 to 1546 days (mean 274). We performed direct immunofluorescence with C3, immunoglobulin M, and immunoglobulin G antibodies on frozen sections of 106 transbronchial biopsies and one wedge biopsy and compared the results with 13 explanted lungs, one donor lung, and two controls. The histologic diagnoses of these 107 biopsies included acute cellular rejection (62, 58% [minimal 23, mild 33, moderate 5, and severe 1]), chronic rejection (eight, 7%), chronic vascular rejection (two, 2%), acute vasculitis (five, 5%), cytomegalovirus pneumonitis (two, 2%), acute pneumonia (two, 2%), acute organizing pneumonia (two, 2%), diffuse alveolar damage (one, 1%), no evidence of rejection or infection (30, 28%), lipoid pneumonia (one, 1%), and inadequate for histologic diagnosis (one, 1%). Eighty-nine of 106 (84%) transbronchial biopsies, the wedge biopsy, and control lungs were satisfactory for direct immunofluorescence, because each contained alveolate lung parenchyma and arterioles or venules. There was no demonstrable immunofluorescence in the wall of the blood vessels or in the lung parenchyma in any case. We conclude that (1) transbronchial biopsies and wedge biopsies provide adequate material to evaluate humoral rejection, and (2) in spite of the large population studied, the satisfactory material obtained, and the wide range of histologic diagnoses, we could not demonstrate the occurrence of humoral rejection in the lung.

Cytologic features of clear cell carcinoma of the urethra and urinary bladder.

Clear cell carcinoma (CCL) arising in the lower urinary tract is unusual and we report the cytohistologic findings of three cases retrieved from our files. All patients presented with bleeding, and the tumors were localized in either the urethra or bladder base. Filter and cytocentrifuge preparations of the urine were studied and all cases displayed numerous scattered aggregates or single tumor cells in an inflammatory background. The enlarged cells had abundant clear, wispy cytoplasm with discrete vacuolation. Hobnail and signet ring cells were apparent. The nuclei had granular to vesicular chromatin with prominent often multiple nucleoli. The tumors were histologically distinctive and typically had a tubulocystic configuration with varying proportions of papillary and diffuse patterns. One patient has died of metastatic cancer and two are presently free of tumor. The cytohistologic features of this cancer are characteristic and from our review we conclude that this lesion can be diagnosed by cytologic means.

[Preliminary study of the safety and immunogenicity of the attenuated VD47/25 strain of camelpoxvirus]. / Etude préliminaire de l'innocuité et du pouvoir immunogène de la souche atténuée VD47/25 de camelpoxvirus.

The safety and immunogenicity of the attenuated VD47/25 strain of camelpoxvirus were tested on 30 camel calves in Mauritania. Post-inoculation clinical symptoms were absent during the 40 days of observation. Serum samples collected during this period showed low levels of neutralizing antibodies (1/4-1/16). In vivo titration of a virulent strain of camelpoxvirus in vaccinated camels and control animals enabled the calculation of the PD50 (50% protective dose) which contained the equivalent of 10(3.7) TCID50 (50% cell culture infective dose). Other studies are still required to determine the dose of this vaccine needed to protect 95% of vaccinated animals.

Unsatisfactory aspirates from fine-needle aspiration biopsies: a review.

While there is much evidence that fine-needle aspiration biopsy (FNAB) is sensitive and specific, there is little information comparing the proportions of unsatisfactory aspirates obtained by clinicians or pathologists. We reviewed 2,199 FNAB reports of superficial lesions. Cases were grouped by organ/site and according to who performed the biopsy. The proportions of unsatisfactory aspirates were computed for clinicians and pathologists. Both groups performed approximately equal numbers of procedures. Overall, 9% of aspirates obtained were unsatisfactory (n = 191). Pathologists had lower proportions of unsatisfactory aspirates in all sites. Of aspirates obtained by clinicians, 14% were unsatisfactory, compared to 3% of those obtained by pathologists (P < .00001). The proportion of unsatisfactory aspirates appears to decrease as physician experience increases, and pathologists may have more experience with FNAB than do clinicians. Other advantages pathologists may have include technique and working with tissue regularly. Both clinicians and pathologists can expect to decrease their proportions of unsatisfactory aspirates by performing FNABs frequently.

The influence of high dietary protein, energy and mineral intake on deficient young camel (Camelus dromedarius)--I. Changes in metabolic profiles and growth performance.

1. The main forage for camels in northern Djibouti (mangrove with Avicennia marina) is very poor in nitrogen and energy. In a trial, 32 young camels (less than 2 years old) were used in four groups of eight each. 2. All the camels received mangrove as basal diet ad lib. 3. After 1 month, the camels received mineral supplementation in copper and zinc (groups 1 and 3) or/and a concentrate rich in protein and energy (groups 2 and 3) or continued with the basal diet (controls). 4. Any supplementation was stopped after 2 months for 1 month. 5. Growth performance was 550 g/day (concentrate-supplemented camels) and 570 g/day (concentrate+mineral-supplemented camels). 6. The growth was negative for the two others groups (-260 g/day). 7. Food intake of mangrove was slightly more important with mineral supplementation only and with mineral+concentrate supplementation. 8. The changes in metabolic profiles have shown an important catabolism in non-supplemented animals, an increase of urea and free fatty acid concentrations in plasma and a decrease of glucose concentrations. 9. Three camels died in the control group with symptoms of starvation and signs of liver damage (increase of liver enzymes glutamate dehydrogenase and gamma-glutamyl transferase).

The influence of high dietary protein, energy and mineral intake on deficient young camel (Camelus dromedarius)--II. Changes in mineral status.

1. Mangrove Avicennia marina is poor in some trace elements such as copper, zinc and manganese. In a trial we used 32 young camels divided into four groups. 2. Groups 1 and 3 were supplemented with copper and zinc in drinking water after 1 month of mangrove feeding. 3. Groups 2 and 3 received concentrate rich in protein and energy. The supplementation was stopped after 2 months. 4. All the camels were deficient in trace elements at the beginning of mineral supplementation. 5. The plasma concentration of copper increased significantly up to normal levels (less than 70 micrograms/100 ml) in energy protein supplemented groups, but the quantity supplied (100 mg of copper sulphate/day) was not sufficient to maintain this level after the end of supplementation. 6. The original zinc deficiency was too severe to observe a significant effect of the mineral supplementation. 7. Calcium, magnesium and phosphorus levels were improved during the supplementation period in protein-energy supplemented groups. 8. A high interaction between mineral absorption and quality of the diet was observed. A well-balanced diet seems essential to avoid deficient mineral status.