Staffan Smeds

Randomized clinical trial of groin hernia repair with titanium-coated lightweight mesh compared with standard polypropylene mesh

Groin hernia repair is one of the commonest operations in general surgery. Existing techniques have very low and acceptable recurrence rates, but chronic pain and discomfort remain a problem for many patients. New mesh materials are being developed to increase biocompatibility, and the aim of this study was to compare a new titanium‐coated lightweight mesh with a standard polypropylene mesh.

Telomerase activity in surgical specimens and fine-needle aspiration biopsies from hyperplastic and neoplastic human thyroid tissues

BACKGROUND Telomerase activity (TA) indicates malignancy, but activated lymphocytes also express TA. Correlation between TA in thyroid tissues and fine-needle aspiration (FNA) samples and knowledge about TA in adjacent tissue are of importance. METHODS The telomeric repeat amplification protocol assay followed by enzyme-linked immunosorbent assay detection was performed on 78 thyroid cases including 53 suspected malignancies, preoperative and perioperative FNA specimens, and adjacent tissue. RESULTS Benign lesions in cancer-suspected cases were TA negative. Eight of 13 papillary (62%) and 4 of 5 follicular (80%) tumors were TA positive (TA+). Lower TA was observed in conventional papillary cancer than in follicular, tall cell variant of papillary and anaplastic cancers. Adjacent tissues with lymphocyte infiltration were TA+ in 9 of 17 cases (53%). Nine of 65 adjacent tissues (14%) were TA+. Three of 6 preoperative and 9 of 11 perioperative FNA samples from malignant tumors corresponded to the tissue TA. CONCLUSIONS; High TA may reflect more severe thyroid cancer. Telomerase activity in FNA biopsies does not add reliable diagnostic information, and presence of lymphocytes can give false-positive results.

The role of aspiration cytology in the management of thyroid nodules

In the hands of an experienced cytologist aspiration cytology is a safe and hitherto the best diagnostic tool in the evaluation of nodular thyroid lesions. In histologically verified case series 50-90% of confirmed thyroid cancers can be detected by aspiration biopsy, the sensitivity being dependent on sampling errors, microscopic misinterpretation and the variation in attitude towards indeterminate diagnosis in the decision for diagnostic surgery. The number of proven benign cases that are correctly identified as such by biopsy varies accordingly and approx. 75% (specificity). In comparison with imaging procedures, including those giving information of functional activity, the combined sensitivity and specificity rates of aspiration cytology come closest to the ideal discriminatory situation. In combination with case history and careful clinical examination, fine needle aspiration cytology is the best guidance for an optimal selection of patients for therapeutic or diagnostic surgery. Future development of sensitive markers for malignant degeneration will probably increase the selective power of this diagnostic technique.

The Influence of a load of L-arginine on serum amino acids and pancreatic apoptosis/proliferation and ATP levels in the rat.

Objectives: Administration of high doses of amino acids like ethionine, methionine, and arginine causes pancreatic tissue damage. The initial mechanism behind these effects is not known. The aim of this study was to show the early effects of a load of l-arginine on programed cell death/proliferation and ATP levels in the pancreas. Methods: We analyzed in rats the effects of intraperitoneal administration of l-arginine on serum amino acids, pancreatic cell apoptosis/proliferation, and ATP levels at 8, 16, and 24 hours. Serum amino acid concentrations were measured with HPLC, tissue ATP was measured fluorometrically, apoptosis was studied with caspase-3 activity and histone-associated DNA-fragments, and proliferation was studied with thymidine autoradiography. Results: After a load of l-arginine, there were initially increased serum levels of l-arginine and l-citrulline, but these fell below control levels after 24 hours as well as amino acids in the glutamate family (ornithine, proline, histidine, and glutamine). Initially, increased ATP levels in the pancreatic tissue returned to control levels at 24 hours. The acinar cells proliferation was suppressed and the apoptosis rate strongly increased at 16 and 24 hours. Pancreatic histology showed vacuole formation in the acinar cells at 8 hours. At 16 hours, there was less vacuolization, but apoptotic bodies were seen, and at 24 hours there was cell degeneration but no necrosis. Conclusions: After a load of l-arginine, amino acid metabolism causes a high ATP production in the pancreatic tissue that may cause mitochondrial initiation of cell death.

Histopathology grading in small brast cancers ≤ 10mm – results from an area with mammography screening

Histopathological malignancy grading ysing the Bloom-Ricardson classification of ductal cancers was performed for 248 invasive ductal breast cancers ≤10mm operated 1978-1985. There weresignificantly more grade 1 lesions in the prevalence screening round. Grade 3 was correlated with aneuploidy, higher S-phase (SPF), and more receptor negative tumours. There were also significantly more positive lymph nodes in grade 3 lesions, 18% compared to 5% and 12% respecrively for grades 1 and 2 (p<0.05). In life table analysis for survival, when the high risk group of grade 3 lesions was compared to the grade 1 and 2 lesions combined, five-year disease-free survival was 84.6% vs. 99.1% (p<0.001).With good training and care from the pathologist, malignancy grading seems useful for prognostication of eventual recurrence and death. In umours 10mm or smaller only grade 3 lesions need to be included in follow-up systems and should probably have adjuvant treatmentMalignancy grading is especially good in small ductal brast cancers whre grading can always be performed while other prognostic determinations are hampered by shortage of material. Lymph node positivity is also low in this groupHistopathological malignancy grading ysing the Bloom-Ricardson classification of ductal cancers was performed for 248 invasive ductal breast cancers ≤10mm operated 1978-1985. There weresignificantly more grade 1 lesions in the prevalence screening round. Grade 3 was correlated with aneuploidy, higher S-phase (SPF), and more receptor negative tumours. There were also significantly more positive lymph nodes in grade 3 lesions, 18% compared to 5% and 12% respecrively for grades 1 and 2 (p<0.05). In life table analysis for survival, when the high risk group of grade 3 lesions was compared to the grade 1 and 2 lesions combined, five-year disease-free survival was 84.6% vs. 99.1% (p<0.001). With good training and care from the pathologist, malignancy grading seems useful for prognostication of eventual recurrence and death. In umours 10mm or smaller only grade 3 lesions need to be included in follow-up systems and should probably have adjuvant treatment Malignancy grading is especially good in small ductal brast cancers whre grading can always be performed while other prognostic determinations are hampered by shortage of material. Lymph node positivity is also low in this group

Microdialysis of 5-S-cysteinyldopa from interstitial fluid in cutaneous human melanoma transplanted to athymic mice

Microdialysis was investigated as a tool for the determination of the extracellular concentration of the pigment metabolite 5-S-cysteinyldopa in human melanoma transplanted to athymic mice. Histology of the tumour with the microdialysis probes in situ showed no tissue damage. With probes equipped with polycarbonate membranes (20 kD) extraction (relative recovery) was approximately 50% at pH 4.0 and flow rates of 1 μl/min, but at pH 7.0 recoveries were markedly lower, particularly from serum. In a first series of human melanomas transplanted to athymic mice low concentrations of 5-S-cysteinyldopa were detected in only two out of ten dialysates and were not detected in the other eight. Utilizing devices constructed for comparison of membrane characteristics in vitro we found about 4-fold higher recoveries with cuprophane and polyamide membranes than with polycarbonate membranes. Therefore newly constructed microdialysis probes (CMA/11) with cuprophane membranes were tested in vitro and gave recoveries of 38–48% from Ringer-Acetate solutions and 22–31% from serum, and the pH effects were low. When these probes were utilized in a second series of melanomas transplanted to athymic mice, 5-S-cysteinyldopa could easily be quantified in 10/10 experiments. A steady-state level of the dialysate 5-S-cysteinyldopa concentration was reached after 45 min.

Revascularisation of human parathyroid tissue transplanted to athymic mice

The revascularisation process of transplanted human normal, hyperplastic and adenomatous parathyroid tissue was analysed at 2 and 4 days and 1, 2, 4, 7 and 12 weeks after transplantation to athymic mice. The transplants were examined by light and electron microscopy, immunohistochemistry and autoradiography. Vessels were detected by monoclonal antibodies specific for mouse and human endothelial cells. Immunohistochemistry demonstrated ingrowth of vessels from the host into the transplant and at one week numerous capillary sprouts were observed in the peripheral parts of the transplants. During the first week, peak levels of proliferation (labelling index) were observed in endothelial cells and capsular fibroblasts, and the proliferative capacity of endothelial cells was most pronounced in adenoma transplants. Fenestrated capillaries were observed in hyperplastic and adenomatous transplants, but not in transplants of normal tissue. In conclusion, revascularisation of transplanted human parathyroid tissue is enabled by ingrowth of vessels from the host into the transplant. The proliferative capacity of endothelial cells is higher and the process of maturation is faster in hyperplastic and adenomatous tissue compared to normal tissue.

A high-sensitivity fluorometric high-performance liquid chromatographic method for determination of glutathione and other thiols in cultured melanoma cells, microdialysis samples from melanoma tissue, and blood plasma.

A high-performance liquid chromatographic method with fluorometric detection is described which is suitable for determination of glutathione in small samples. Reduced glutathione (GSH) and total glutathione obtained as GSH after reduction with glutathione reductase is derivatized with N-(7-dimethylamino-4-methyl-3-coumarinyl) maleimide (DACM) and subjected to chro-matography. The detection limit for the GSH-DACM derivative was 5–10 fmol/injection, and analytical recovery was quantitative. The method is suitable for determination of both reduced and total glutathione in samples from microdialysis of melanoma tumours, and cysteine can be quantified in the same chromatogram. Application is shown also for glutathione determinations in cultured melanoma cells, melanoma homogenates and plasma.

Growth and function of human parathyroid tissue transplanted to athymic mice

The morphology, cell proliferation and function of transplanted normal, hyperplastic and adenomatous human parathyroid tissue was studied after transplantation to athymic mice. The iPTH was evaluated in relation to morphology. Human parathyroid tissue collected during surgery for hyper-parathyroidism was implanted subcutaneously into athymic mice (nu/nu-BALB/cA) and was analysed 1, 4, 7 and 12 weeks after transplantation. The transplants were examined by light and electron microscopy and by autoradiography after continuous infusion of 3H-thymidine. The relative amount of viable tissue was evaluated using a computer image analysing programme. Graft function was evaluated by measuring human iPTH in mouse serum. A transplant take ratio of 93% was observed. The proliferation rate in adenoma grafts at 12 weeks after transplantation was five and fifteen times that observed in normal and hyperplastic transplants, respectively. In normal and adenoma groups, a continuous increase in iPTH concentrations was observed, but in trie hyperplastic group the iPTH remained on the same level. The secretion of iPTH in relation to the amount of transplanted tissue and the fraction of viable tissue was at the same level at 12 weeks in normal and adenomatous grafted animals. In conclusion, human parathyroid tissue was successfully transplanted and maintained its original structure. The growth potential, but not the iPTH secretion, was significantly higher in adenoma grafts compared to grafts from hyperplastic and normal glands.

Plasma membrane shedding and colloid vacuoles in hyperactive human thyroid tissue

SummaryThe ultrastructural appearance of colloid vacuoles, considered to be a typical sign of hyperactivity in the human thyroid gland, was studied in human thyroid tissue transplanted to nude mice and in human thyroid tissue fixed directly after surgical removal in patients with thyrotoxicosis. Transplanted normal thyroid tissue and toxic diffuse goiter (TDG) tissue was fixed by vascular perfusion with glutaraldehyde 5 or 12 weeks after transplantation. Light microscopic quantification showed that daily injections for 2 weeks of a gamma globulin fraction of patient sera containing thyroid-stimulating immunoglobulins (TSI) greatly increased the number of colloid vacuoles in both types of transplants. The vacuoles were mainly located in the periphery of the follicle lumen, giving the colloid a scalloped appearance. Electron microscopy of TSI-exposed tissue revealed, in addition to colloid vacuoles, the presence of large amounts of membrane material in the follicle lumen. Only sparse amounts of intraluminal membrane material were present in controls. The colloid vacuoles were almost invariably associated with such membrane material, which lined the border between the vacuole and the surrounding colloid. The intraluminal material consisted of spherical and elongated formations, each structure limited by a triple-layered membrane and often containing a dense interior. The elongated structures were often of the same dimensions as microvilli. The apical surface of follicle cells in TSI-exposed tissue expressed numerous microvilli, of which many showed a similar dense interior as the intraluminal membrane structures. The intraluminal membranes frequently showed, like the apical plasma membrane of the follicle cells, a positive reaction for peroxidase. Organelles, such as mitochondria, lysosomes or rough endoplasmic reticulum, were not encountered among the intraluminal membrane structures. These observations indicate that the intraluminal membrane material is derived from the apical plasma membrane of the follicle cells, presumably by shedding of microvilli. A similar association between colloid vacuoles and membrane material was also found in thyroid tissue from patients with thyrotoxicosis fixed directly at operation. It is suggested that the presence of membrane material in the follicle lumen precipitates the formation of colloid vacuoles in hyperactive thyroid tissue. The possible involvement of intraluminal membrane material in the development of microsomal autoantibodies in Graves’ disease, i.e. exposure and presentation of thyroid microsomal antigen (identical to thyroperoxidase) to the immune system, is discussed.