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Biomedical subjects

J A Hanson

Publications and source records attributed to J A Hanson.

At least 55 records · Page 3Linked to original sources

Determination of radiosensitivity in established and primary squamous cell carcinoma cultures using the micronucleus assay.

In this study, the cytokinesis-block micronucleus assay (CBMN) was used to measure radiosensitivity in three established cell lines (SCC-61, V175 and V134) and 10 primary cell cultures of squamous cell carcinoma (SCC) of the head and neck. Assessment involved optimisation of the assay to determine cytochalasin-B (CB) concentration and sampling time postirradiation. A much closer correlation between dose-response data measured in the clonogenic and micronucleus assays was found when the micronucleus assay was performed under standardised conditions for each cell line (2 micrograms/ml CB: 48 h postirradiation) instead of predetermined optimised assay conditions. This indicates that, for these SCC cell lines, the CBMN assay may be able to predict in vitro radiosensitivity. To be of clinical use in predicting radiosensitivity, the CBMN assay also needs to be evaluated with primary cell cultures. In this study, no relationship between micronucleus frequency at 2 or 6 Gy and patient clinical outcome 12 months following surgery and radiotherapy was seen. Similarly, no association between patient outcome and tumour stage, nodal stage and histology was observed. These CBMN assay data from the primary cell cultures are presently inconclusive as a measure of patient tumour radiosensitivity.

Carcinoma, Squamous Cell↗

The influence of experience on the reliability of goniometric and visual measurement of forefoot position.

Goniometric measurement of forefoot position relative to the rearfoot is a routine procedure used by rehabilitation specialists. This measurement is also frequently made by visual estimation. The influence of tester experience on the reliability of these two techniques at the forefoot is unknown. The purpose of this investigation was to directly examine the reliability of goniometric and visual estimation of forefoot position measurements when experienced and inexperienced testers perform the evaluation. Two clinicians (> or = 10 years experience) and two physical therapy students were recruited as testers. Ten subjects (20-31 years old), free from pathology, were measured. Each foot was evaluated twice with the goniometer and twice with visual estimation by each tester. Intraclass correlation coefficient (ICC) and coefficients of variation method error were used as estimates of reliability. There was no dramatic difference in the intratester or intertester reliability between experienced and inexperienced testers, regardless of the evaluation used. Estimates of intratester reliability (ICC 2,1), when using the goniometer, ranged from 0.08 to 0.78 for the experienced examiners and from 0.16 to 0.65 for the inexperienced examiners. When using visual estimation, ICC (2,1) values ranged from 0.51 to 0.76 for the experienced examiners and 0.53 to 0.57 for the inexperienced examiners. The estimate of intertester reliability [ICC (2,2)] for the goniometer was 0.38 for the experienced examiners and 0.42 for the inexperienced examiners. When using visual estimation, ICC (2,2) values were 0.81 for the experienced examiners and 0.72 for the inexperienced examiners. Although experience does not appear to influence forefoot position measurements, of the two evaluation techniques, visual estimation may be the more reliable.

Adult↗

Magnetic resonance imaging of adrenocortical adenomas in childhood: correlation with computed tomography and ultrasound.

There are few descriptions of the magnetic resonance (MR) appearance of hyperfunctioning adrenocortical tumours, particularly those occurring in childhood. We studied five patients, two girls and three boys, aged 6-14.3 years, presenting with clinical syndromes of adrenocortical hyperfunction. The diagnoses were Cushing's syndrome (n = 2), virilisation (n = 2), and Conn's syndrome (n = 1). Biochemical features suggested an adrenal lesion in each case. MR and ultrasound were performed in all five cases, with CT in four. Each patient had a functional adrenal tumour secreting either cortisol, androgens or aldosterone alone, or a combination of cortisol, androgens and oestradiol. The histological diagnosis was adenoma in four cases and tumour of indeterminate nature in one case. MR clearly showed the tumours (diameter 1.0-7.5 cm), all the lesions being of high signal intensity relative to liver on T2-weighted sequences. CT revealed an adrenal mass in each of the four patients scanned, three of which enhanced after intravenous contrast medium injection. The multiplanar imaging of MR allowed better distinction from adjacent structures and also demonstrated an unenlarged contralateral adrenal gland. In the patient with a 1-cm Conn's adenoma the lesion was more easily seen on MR than CT. Ultrasound showed the four larger tumours but was unable to visualise the contralateral adrenal or the Conn's adenoma. In conclusion, the MR appearances of four adrenocortical adenomas and one indeterminate tumour in children are described. MR has been found to be at least equal to CT in the detection of these tumours, with some possible advantages. Both techniques are superior to ultrasound.

Adenoma↗

The role of computed tomography in evaluation of subchondral osseous lesions in seven horses with chronic synovitis.

Seven horses with severe, persistent lameness of sudden onset were evaluated with scintigraphy and/or computed tomography. The lameness was localised to the front fetlock joint in 2 horses and to the tibiotarsal joint in 5 horses. Five of the horses had a history of intra-articular injections of the involved joint prior to presentation. All horses had effusion of the affected joint and were positive to flexion tests. Intraarticular anaesthesia eliminated or improved the lameness in 4 cases and a nerve conduction block proximal to the affected joint improved the lameness in another. Cytology examination of fluid from affected joints identified normal joint fluid (one horse) or elevations in nucleated cell counts of 0.9 x 10(9)/l-36.8 x 10(9)/l and total protein 20-42 g/l (6 horses). The joint fluid of 2 of these horses cultured positive for bacteria. Initial radiographs were either normal (4 cases) or the changes seen were not sufficient to explain the degree of lameness. In the 6 cases where scintigraphy was performed, intense focal isotope uptake was found in the suspected region, which corresponded to the proximal portion of the first phalanx (2 cases), distal tibia (2 cases), or talus (3 cases). Computed tomography (CT) was performed because occult fracture or osteomyelitis was suspected; and knowledge of the precise anatomical location of the lesion was considered necessary to assess the need for surgery and to plan the surgical approach. Hypodense focal lesions with hyperdense haloes were found in the subchondral bone deep to the sagittal groove of the first phalanx (P1) (2 cases) in the cochlea of the distal tibia (2 cases), and in the intertrochlear portion of the talus (3 cases). Communication between the lesion and the joint space was demonstrated by CT in 5 cases. Post mortem examination of one case revealed synovitis and a chronic bone abscess (Brodie's abscess) communicating with the joint space.

Animals↗

Myasthenia gravis presenting with stridor.

The case is described of a 72 year old woman who presented with a two year history of exertional stridor in whom the diagnosis of myasthenia gravis was delayed. Although an uncommon cause, myasthenia gravis should be included in the differential diagnosis of stridor.

Aged↗

The sensitivity of chronic lymphocytic leukaemia lymphocytes to irradiation in vitro.

The inhibition of [3H]-thymidine incorporation into the DNA of mitogen-stimulated chronic lymphocytic leukaemia lymphocytes by chlorambucil or gamma-irradiation in vitro was measured in a series of patients, some of whom were untreated, some treated and some who were showing resistance to first-line or second-line treatment. There was evidence of resistance to irradiation developing in parallel with that to chlorambucil. The resistance to chlorambucil in chronic lymphocytic leukaemia (CLL) is not necessarily due to altered drug transport or metabolism but to a more fundamental process affecting DNA damage.

Antineoplastic Agents, Alkylating↗

Determination of radiation-induced damage in lymphocytes using the micronucleus and microgel electrophoresis 'Comet' assays.

DNA damage assays may be useful as rapid predictors of normal tissue radiosensitivity in clinical samples. We measured in vitro radiation-induced (2 Gy) damage to lymphocytes from cancer patients and normal healthy donors using both the micronucleus and microgel electrophoresis (Comet) assays simultaneously. For damage assessment, there was a good correlation (P < 0.001) between the mean comet lengths and the fraction of cells with comets. There was no correlation with initial damage, determined as the proportion of cells within a sample that formed comets, in comparison with the mean frequency of micronuclei per binucleate cell. However, there appeared to be an association between the determination of repair proficiency in the Comet assay and the mean frequency of micronuclei per binucleate cell in lymphocytes from cancer patients.

DNA↗

The micronucleus assay: an evaluation of its use in determining radiosensitivity in vitro.

The cytokinesis-block micronucleus assay was used to measure radiosensitivity in vitro in a panel of seven cell lines. Six of these cell lines were used to study the major parameters of this assay. We observed varying sensitivities following cytochalasin-B exposure. Treatment with 1 microgram/ml cytochalasin-B for 24 h reduced cell survival in four of the six cell lines by > 60%. Cytochalasin-B concentration and post-irradiation culture time were both found to influence cell-response. In three cell lines (V39, V134 and HX142), a decrease in cytochalasin-B concentration (2-0.5 microgram/ml) resulted in an increase in the frequency of radiation-induced micronuclei per binucleate cell. In other cell lines, either the opposite (V7M, CHO-K1) or no effect (WiDr) was seen. A linear dose-response was observed between induced damage expressed as the frequency of micronuclei and radiation dose in all but one melanoma (V39) cell line. Evidence for radiation-induced division-delay, with the maximum frequency of binucleation in irradiated cultures occurring 24-48 h after that of controls, was only seen in two cell lines. Of particular note, and in contrast to some other published reports, was the lack of a general correlation between cell-response measured in the clonogenic and the cytokinesis-block micronucleus assays. Consideration of lethal lesions, determined from the clonogenic dose-response curve, with respect to micronucleus frequency showed a complex relationship, with one micronucleus per binucleate cell corresponding to a wide range of lethal lesions depending on the cell line.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Aortic root thrombus causing stroke in a patient with systemic lupus erythematosus.

We report on a 24 year old woman with systemic lupus erythematosus (SLE) who presented with a cerebrovascular accident affecting her right side. Unusually, this was associated with an aortic root thrombus with absence of any commonly recognised thrombophilic features. Following removal of the thrombus and anticoagulation, she made a good recovery. The relationship between valvular problems and cerebral lesions in SLE is discussed.

Adult↗

Organoapatites: materials for artificial bone. II. Hardening reactions and properties.

This article reports on chemical reactions and the properties they generated in artificial bone materials termed "organoapatites." These materials are synthesized using methodology we reported in the previous article of this series. Two different processes were studied here for the transition from organoapatite particles to implants suitable for the restoration of the skeletal system. One process involved the hardening of powder compacts by beams of blue light derived from a lamp or a laser and the other involved pressure-induced interdiffusion of polymers. In both cases, the hardening reaction involved the formation of a polyion complex between two polyelectrolytes. In the photo-induced reaction an anionic electrolyte polymerizes to form the coulombic network and in the pressure-induced one, pressure forms the complex by interdiffusion of two polyions. Model reactions were studied using various polycations. Based on these results the organoapatite selected for the study was that containing dispersed poly(L-lysine) and sodium acrylate as the anionic monomer. The organomineral particles can be pressed at room temperature into objects of great physical integrity and hydrolytic stability relative to anorganic controls. The remarkable fact about these objects is that intimate molecular dispersion of only 2-3% by weight organic material provides integrity to the mineral network in an aqueous medium and also doubles its tensile strength. This integrity is essentially nonexistent in "anorganic" samples prepared by the same methodology used in organoapatite synthesis. The improvement in properties was most effectively produced by molecular bridges formed by photopolymerization. The photopolymerization leads to the "hardening" of pellets prepared by pressing of organoapatite powders. The reaction was found to be more facile in the microstructure of the organomineral, and it is potentially useful in the surgical application of organoapatites as artificial bone.

Apatites↗

Organoapatites: materials for artificial bone. III. Biological testing.

This article reports on the in vivo testing of new artificial bone materials we have termed "organoapatites." These materials consist of mineral networks in which organic polymers are intimately dispersed by nucleation and growth of apatite crystals from a mother liquor containing the organic substances. Organoapatites were tested as implants in adult canine cortical bone for periods in the range from 12-35 weeks and fluorochromes were used in the model to investigate the kinetics of bone growth or repair. The analysis of histological samples was carried out using histomorphometric methods as well as fluorescence microscopy. Results showed excellent apposition of poly(amino acid) organoapatites with mineralized bone and fibrous encapsulation when a synthetic polyelectrolyte was the only organic component. This observation suggests that the molecularly dispersed organic dopant amounting to only 2-3% by weight of the microstructure can play a critical role in the tissue response to the implant. Relative to apatite controls, organoapatites were also found to have greater resistance to fragmentation in vivo and those containing amino acid units revealed interfacial bioerosion accompanied by regeneration of mineralized tissue. Design of organoapatite compositions and microstructures may therefore be useful in achieving the specific rate of biological response which is clinically desired.

Animals↗

Calibration of dual-energy x-ray absorptiometry for bone density.

Bone mineral content (BMC, g) using DEXA (Lunar DPX) was measured on known hydroxyapatite samples in a water bath in the presence of uniform and nonuniform covering of fat-equivalent materials. Selective placement of paraffin over bone had a greater effect than lard in reducing apparent BMC, and polycarbonate plastic had a lesser effect. Measured BMC was 100.1 +/- 1.1% of actual hydroxyapatite weight when (1) fat over bone was about twice the mass of hydroxyapatite, and (2) the surrounding soft tissue was 15-30% fat. There was a linear relationship between observed and expected BMC, area (cm2), and bone mineral density (BMD, g/cm2) measured on an aluminum phantom using either the Lunar DPX or the Hologic QDR-1000. The measured area with the two densitometers was identical, but BMC differed. For both an anthropomorphic phantom and human subjects, use of a constant-threshold (0.2 g/cm2) edge-detection algorithm excluded less low-density bone from the transverse processes than the standard DPX edge-detection algorithm. Differences in edge detection could influence the results obtained with phantoms and in vivo and make system intercomparison difficult.

Absorptiometry, Photon↗

DEXA measurement of spine density in the lateral projection. I: Methodology.

Bone mineral content and bone mineral density (BMC in g and BMD in g/cm2) were measured using dual-energy X-ray absorptiometry (DEXA). DEXA scans in the lateral decubitus position required about 12 minutes for the L2-L4 sequence at 0.75 mA (dose 5 mrem) and 4 minutes at 4.75 mA (7 mrem). The former scans were done with the Lunar DPX densitometer and the latter with the Lunar DPX-L. One test of the algorithms used for measurement is the equality of BMC in both AP and lateral projections. BMC in the lateral projection averaged about 1% lower than in the AP projection in phantoms and for L2 + L3 in 8 subjects, but the difference was not significant. Additional tests were done on the effects of tissue thickness and position from the tabletop. There was little or no influence of tissue thickness from 18 to 30 cm on BMD results, but there was a small influence of thickness below 18 cm (0.01 g/cm2; P = 0.01) and of distance from the tabletop at extremes of positioning (0.02 g/cm2; P = 0.06). The precision in vivo was similar for both 4- and 12-minute scans; the standard deviation of repeat measurements was about 0.02 g/cm2, which was about 2% relative to the mean BMD for a region within the vertebral body. The latter region included half the BMC of the body, or 24% of the entire vertebra. Results of 4-minute scans on the DPX and 12-minute scans on the DPX-L in 9 subjects were highly correlated (r = 0.98; P less than 0.001).

Absorptiometry, Photon↗

In vitro chemosensitivity testing in chronic lymphocytic leukaemia patients.

Twenty-nine samples from eighteen patients with chronic lymphocytic leukaemia (CLL) were used in a direct comparison of in vitro response to chlorambucil measured in a metabolic (MTT) and dye exclusion (D.Ex) assay. Reduced ability to produce formazan corresponded to a reduced number of dye-excluding viable cells and a significant correlation was found between dose-response measured in the two assays. Initial low absorbance values obtained with untreated control cells in the MTT assay were effectively overcome by increasing both the cell seeding density and MTT exposure time with the consequent increase in assay sensitivity. The MTT assay provided qualitatively similar dose-response data to that obtained in the D.Ex assay. A wide range in in vitro response was seen for both pretreatment and treatment patient groups. In vitro dose-responses were seen to coincide with decreasing or stable white cell counts, taken around the time of sampling, in samples from 4 patients. Less marked dose-responses were observed for 2 patients considered clinically resistant to chlorambucil. The MTT assay would seem, therefore, to be applicable to in vitro assay of cell response in CLL.

Chlorambucil↗

Fibrosing cholehepatitis in broiler chickens induced by bile duct ligations or inoculation of Clostridium perfringens.

The pathogenesis of fibrosing hepatitis causing condemnations in broiler chickens was investigated. Three to four week old broilers were inoculated via the hepatoenteric bile duct with saline washed suspensions of Clostridium perfringens (10(7) and 10(8) organisms). In another group of broilers, both bile ducts were ligated. The sequential development of liver and gall bladder lesions was studied at intervals ranging from 1-28 days postsurgery. The lesions were similar in both experiments in that the liver became mottled and swollen by five to seven days. Fibrinoid necrosis, heterophil and lymphocyte infiltration, bile duct hyperplasia and fibrosis with reticulin fiber proliferation occurred. By 14-17 days, the liver was enlarged, tan colored and firm with red and white foci. By 28 days, bile duct proliferation and fibrosis were massive with only a few hepatocytes remaining. The liver capsule was not involved. Jaundice was not present but the birds with ligated bile ducts excreted intensely yellow stained droppings after six to seven days. The gall bladder in inoculated birds was edematous and distended with flocculent or inspissated material. Clostridium perfringens was reisolated from gall bladder and/or liver of inoculated birds up to 28 days postsurgery. It is suggested that this organism plays a role in the pathogenesis of fibrosing cholehepatitis by inducing septic intrahepatic cholestasis.

Animals↗

A novel dye exclusion assay for measurement of cell response following in vitro exposure to radiation or drugs.

A novel dye exclusion assay based on differential staining of fixed cells has been evaluated using the MOLT4 (T-lymphoblastic leukaemia) and DAUDI (Burkitt lymphoma) cell lines. Reproducible differential staining was found with fixed samples kept for up to 2 months. A small increase (less than 13%) in the proportion of stained cells was observed over the first 30 days in fixative. Reproducible dose-response data were obtained with cells treated with radiation or drugs and assayed after 4 days in culture. This ability to fix cells prior to measurement of viability is of general use particularly where time constraints prevent haemocytometer counting. More specifically, it may have potential use in the field of chemosensitivity testing particularly where large sample numbers require rapid processing.

Alcian Blue↗