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Biomedical subjects

W J Kort

Publications and source records attributed to W J Kort.

At least 19 recordsLinked to original sources

A microchip implant system as a method to determine body temperature of terminally ill rats and mice.

In a series of experiments, Klebsiella pneumoniae was inoculated intratracheally into rats and mice, and the temperature of the animals was recorded twice daily using microchip transponders. Transponders are interrogated by radio frequencies and were implanted either subcutaneously or intraperitoneally. The microchip temperatures were compared with rectal temperatures taken at the same time. The purpose of the experiments was (a) to investigate the practicability and reliability of the ELAMS for temperature recording; (b) to compare values given by subcutaneously or intraperitoneally implanted transponders with rectal temperatures; and (c) to determine a 'temperature-cut-off point' as an alternative for 'death of the animal' as an end point for the experiment. The results showed that the ELAMS was easy to operate and no important drawbacks in the use of the system were observed. The temperatures generated by the transponders implanted subcutaneously and intraperitoneally did not differ significantly from rectal temperatures. In two out of three experiments on rats, it was shown that when the temperatures reached values below 36 degrees C, the median survival time of the animals was 24 h. In the one experiment on mice the same median survival time was observed at 36 degrees C. In one experiment using rats, however, the disease was so acute that death occurred before any temperature drop was seen. The results show that when a 36 degrees C cut-off point is used instead of the time of death in this particular animal model, the statistical analysis was not altered, but that it would spare animals further suffering for approximately 24 h. The argument that measuring body temperature is a laborious job and stressful to the animals is overcome when the ELAMS system is used.

Animals↗

The effect of cigarette smoking on the survival of free vascularized and pedicled epigastric flaps in the rat.

Microsurgeons suspect that cigarette smoking reduces the survival of free vascularized flaps and replantations, but this has never been proven. This experimental study investigates the effect of smoking on free-flap survival. A fasciocutaneous epigastric flap was used in 30 rats as a free flap and in 30 rats as a pedicled flap. Of each group, 10 rats were smoked 6 weeks before and 2 weeks after surgery, 10 rats were smoked only 6 weeks before surgery, and 10 rats underwent the sham smoking procedure. Also, a distally based dorsal skin flap was cut in all rats, representing a random vascularized flap. Vitality and size of both flaps and patency of the vascular anastomoses were assessed 14 days after surgery. The epigastric flaps were monitored by laser Doppler flowmetry and thermometry during the experiment. Survival of the free vascularized epigastric flaps was significantly lower in smoking rats. All pedicled flaps except one survived. The epigastric flaps only necrosed or survived completely, exactly correlating to the patency of the vascular anastomoses. The mean surviving area of the dorsal flaps was best for nonsmoking rats, worse for only preoperatively smoking rats, and worst for preoperatively and postoperatively smoking rats. The differences were statistically significant. Postoperative laser Doppler flow differed significantly between surviving and dying flaps, affirming the value of laser Doppler flow monitoring in microvascular surgery. In conclusion, this study proves that smoking of cigarettes is detrimental to the survival of free vascularized flaps.

Anastomosis, Surgical↗

Adjuvant intraoperative photodynamic therapy diminishes the rate of local recurrence in a rat mammary tumour model.

The use of photodynamic therapy (PDT) as an adjunct to curative tumour resection was investigated in a tumour recurrence model, using rat mammary adenocarcinoma BN472. Tumours were inoculated subcutaneously in 60 animals and resected after 21 days of growth. Immediately after removal, the operation site was exposed to 320-450 nm light of 0.1 W cm-2 and 60 J cm-2 after photosensitisation with either Photofrin (5 mg kg-1 i.v. 48 h before illumination) or 5-aminolaevulinic acid (ALA) (2 mg ml-1 in drinking water for 9 days). Porphyrin concentrations were measured in tissue samples. After 28 days, animals treated with adjunctive PDT had a significantly longer tumour-free interval than controls (P < 0.01); median 25 days (Photofrin), 18 days (ALA), 14 days (controls). Moreover, in the PDT groups significantly fewer rats had lymph node metastasis. A prophyrin concentration ratio between tumour and mammary tissue of 2:1 was found after Photofrin and 4:1 after ALA. The results indicate that adjuvant intraoperative PDT may be a safe and effective method of destroying residual tumour, thereby preventing locoregional tumour recurrence.

Adenocarcinoma↗

Interstitial Nd:YAG laser coagulation with a cylindrical diffusing fiber tip in experimental liver metastases.

Interstitial laser coagulation as a means of destructing hepatic metastases was investigated. Colon carcinoma CC531 was implanted in the liver of 42 Wag/Rij rats; 20 days later, tumors (5.5 +/- 0.2 mm) were exposed to 1,064 nm Nd:YAG laser light at 4 W/cm and either 600, 1,200, 2,400, 3,400, or 4,800 J/cm from a 0.5 cm Helioseal coated cylindrical diffuser. Temperature and fluence rate were measured at the tumor boundary. Lesions were studied on day 2 and 36 posttreatment by light microscopy. Tumor proliferative activity was assessed by bromodeoxyuridine incorporation. Liver damage and function were determined by serum liver enzymes and antipyrine clearance. Fluence rate increased during laser treatment up to 170%; mean temperature increased logarithmically up to 69.7 degrees C. Short-term light microscopy showed coagulation necrosis of 7-11 mm without charring. Lesion size and liver enzymes increased logarithmically with laser energy applied. No deterioration in liver function was found. At 4,800 J/cm complete tumor remission occurred in three of four animals. This study shows the ability of interstitial laser coagulation to produce selective destruction of colonic tumor within the liver.

Adenocarcinoma↗

Current status of photodynamic therapy in oncology.

Photodynamic therapy (PDT) is a cancer treatment based on the accumulation in malignant tissue of a photosensitiser with low systemic toxicity. Subsequent illumination induces a type II photochemical reaction with singlet oxygen production that results in destruction of biomolecules and subcellular organelles. The first full clinical report of PDT dates from 1976. Haematoporphyrin derivative, a complex mixture of porphyrins, was initially used as a photosensitiser. An enriched fraction (porfimer sodium) is now the most commonly used clinical agent. After systemic administration porphyrins bind to albumin and lipoproteins. Accumulation occurs mainly in tumours and organs of the reticuloendothelial system. The light of an argon-dye laser can be tuned to the appropriate wavelength and delivered either superficially, interstitially or intraluminally. Light distribution can be assessed by using a radiation transport model and tissue optical properties, or direct measurement with light detectors. The effects of PDT depend in a complex way on: characteristics, tissue concentration and localisation of the photosensitiser; the target tissue optical properties and oxygenation; activation wavelength, power density and treatment regimen. Future research is directed towards: better photosensitisers (i.e. phthalocyanines, chlorins or protoporphyrin IX endogenously produced from 5-aminolevulinic acid); improved light generation and delivery; and combination with hyperthermia, chemotherapy, radiotherapy or surgery. Adjuvant intraoperative PDT is a promising approach to destroying residual tumour after surgery.

Animals↗

Water-jet-cooled Nd:YAG laser coagulation of experimental liver metastases: correlation between ultrasonography and histology.

To establish the value of ultrasonography in imaging laser coagulation of tumor and surrounding tissue, the relation between measurements on ultrasound and histology was determined in a rat tumor model. A piece of colon carcinoma CC531 was implanted in the liver of 21 Wag/Rij rats; 20 days later, tumors (mean diameter 5.3 mm) were treated with a water-jet-cooled Nd:YAG laser at 10 W and either 150 J, 300 J, 600 J, 1,200 J, 1,700 J, or 2,400 J. Ultrasonography was done just pre- and immediately post-laser treatment. The animals were sacrificed and livers removed for light microscopical evaluation. Depth and width of coagulation were measured directly on ultrasound, and on histological samples by computer-assisted image analysis. Laser treatment did not change the echogenic aspect of the tumor on ultrasound. However, liver damage appeared hypoechoic compared to normal liver. A significant correlation was found between the total size of the lesion on ultrasound and histology (P = 0.015, r = 0.57 for depth; P = 0.012, r = 0.58 for width), suggesting that laser induced tumor destruction may be derived from the amount of surrounding hepatic damage on ultrasound.

Adenocarcinoma↗

Transient protection of incomplete colonic anastomoses with fibrin sealant: an experimental study in the rat.

Fibrin glue has been used as a protective seal in normal and high-risk anastomoses to prevent leakage. The influence of fibrin adhesive on the healing colonic anastomosis in a control and high-risk model was tested. Resection and anastomosis of the left colon was performed in rats. In group Ia an end-to-end anastomosis was constructed with 12 7-O polypropylene sutures; in group Ib the anastomosis was sealed with fibrin adhesive. In group II an incomplete anastomosis was constructed with only 4 sutures at 90 degrees, therefore potentially leaking. In group IIb additional sealing with fibrin glue was performed. On Days 2, 4, and 7 body weight, adhesion formation, anastomotic bursting pressure, and collagen concentration were measured. The results showed increased adhesion formation after fibrin sealing. The anastomotic bursting pressure of incomplete anastomoses showed a significant increase after sealing on Day 2 only; on Day 4 and 7 no differences were found. Sealing of control anastomoses caused lower bursting pressures on Day 4. Collagen concentration is significantly reduced after fibrin sealing of normal anastomoses. We conclude that fibrin sealing of control anastomoses inhibits wound healing. Incomplete anastomoses are temporarily protected by fibrin glue sealing. Finally, fibrin sealing of the colon wound does not prevent adhesion formation.

Anastomosis, Surgical↗

Effect of fibrin sealant on the integrity of colonic anastomoses in rats with faecal peritonitis.

OBJECTIVE: To assess the influence of fibrin adhesive on the healing of colonic anastomoses in rats with and without faecal peritonitis. DESIGN: Controlled study. SETTING: Laboratory for experimental surgery, Erasmus University Rotterdam, The Netherlands. MATERIAL: 120 male Wag/Rij rats. INTERVENTIONS: All rats had a single layer end-to-end anastomosis fashioned with 7/0 polypropylene. Faecal peritonitis was then induced in half of the rats by placement of 200 mg powdered autoclaved rat faeces in the peritoneal cavity near the anastomosis. Rats were allocated to one of four groups (n = 30 in each): 1--control; 2--additional sealing with fibrin glue; 3--peritonitis alone; and 4--peritonitis with fibrin glue. MAIN OUTCOME MEASURES: Body weight, adhesion formation, anastomotic bursting pressure and collagen concentration around the anastomosis on days 2, 4, and 7 in 10 rats from each group. RESULTS: 11 rats died of peritonitis before the experiment was completed. Peritonitis caused increased formation of adhesions and abscesses, with or without fibrin sealant. Bursting pressure at the anastomosis was significantly reduced in peritonitis compared with controls on days 4 and 7, and this was not prevented by fibrin. Sealing of anastomoses resulted in lower bursting pressures on day 4 in control animals. Collagen concentration was significantly reduced in peritonitis with or without fibrin sealant on days 4 and 7, and after fibrin sealing of control anastomoses. CONCLUSION: Faecal peritonitis reduced mechanical strength and collagen concentration of colonic anastomoses, and this was not prevented by additional sealing of the anastomosis with fibrin sealant.

Anastomosis, Surgical↗

Effect of antibiotics in fibrin sealant on healing colonic anastomoses in the rat.

In 90 rats a colonic anastomosis was constructed with 12 interrupted 7/0 polypropylene sutures. Group 1 (n = 30) served as a control group. In group 2 (n = 30) the anastomosis was sealed with fibrin adhesive and in group 3 (n = 30) a mixture of fibrin, clindamycin and cefotaxime was used. On days 2, 4 and 7, ten animals in each group were killed. Adhesion formation was significantly increased in groups 2 and 3 compared with the control group. On day 2 the anastomosis was significantly stronger after sealing with antibiotic-fibrin mixture. On day 4 the bursting pressure in group 2 was significantly lower than in groups 1 and 3. At the same time the concentration of hydroxyproline was significantly reduced in group 2, but not in group 3. The addition of antibiotics prevents the negative effect of fibrin adhesive on the healing colonic anastomosis and contributes to a stronger anastomosis on day 2 after operation.

Anastomosis, Surgical↗

A new liver-tumor model in the rat.

A new tumor model for testing and comparing the effect of different forms of treatment on liver tumors is described. Single tumors were induced in the liver of rats by the implantation of small pieces taken from a subcutaneously growing tumor. Tumor growth was determined by measuring the weight of the implanted tumors after the animals had been killed. In this way, weight curves for treated and untreated tumors could be defined. The weight of untreated liver tumors increased exponentially with time (tumor weight in milligrams = 1 + e(t-0.31)/5). In addition, tumor growth defined as the geometric mean of three perpendicular diameters was determined. Tumor-diameter curves showed a linear increase with time in the untreated groups (tumor diameter in millimeters = 0.4 t + 1.90). The model was tested by assessing the effect of intraperitoneally injected cisplatin. The dose chosen produced a marked delay in tumor growth. On the basis of the weight gain shown by the treated animals and tumor growth delay, a therapeutic index can be defined, thus enabling to compare quantitatively different forms of treatment according to their antitumor effect and toxicity.

Adenocarcinoma↗

Effects of long-acting somatostatin analog (SMS 201-995) on eicosanoid synthesis and survival in rats with acute necrotizing pancreatitis.

The effects of a long-acting somatostatin analog (SMS 201-995) were studied in an established model of acute necrotizing pancreatitis in rats. SMS 201-995, when given prior to induction of pancreatitis, decreased the mortality rate from 100% to 40% (P = 0.0001). When treatment was given after induction of pancreatitis, the mortality rate was 75% (P = 0.2). Administration of SMS 201-995 did not influence the serum concentrations of amylase markedly, but the lipase levels were significantly lowered (P less than 0.05). The low levels of serum insulin and the glucose level in whole blood were not influenced. The volume of ascitic fluid was reduced (P less than 0.01). Moreover, less peritoneal fat necrosis was seen, suggesting a reduction in toxic factors in the ascitic fluid. Treatment with SMS 201-995 prior to induction of pancreatitis caused a significant increase in the levels of circulating 6-keto-PGF1 alpha, the stable metabolite of prostaglandin I2 (P less than 0.01). The levels of thromboxane B2 and prostaglandin E2 did not change significantly. The present data support the hypothesis that SMS 201-995 is an activator of prostaglandin I2, thereby modifying the course of the disease.

6-Ketoprostaglandin F1 alpha↗

Healing of ischemic colonic anastomosis: fibrin sealant does not improve wound healing.

Fibrin adhesives have been advocated as a protective sealant in high-risk colonic anastomoses to prevent leakage. To assess the effect of fibrin glue sealing on the healing ischemic anastomosis, we compared the healing of sutured colonic anastomoses in the rat, with and without fibrin adhesive (Groups IA and IB), and ischemic anastomoses with and without fibrin adhesive (Groups IIA and IIB). On days two, four, and seven, 10 animals in each group were sacrificed. Adhesion formation was scored, and the in situ bursting pressure was measured. The collagen concentration and degradation were estimated by measuring hydroxyproline. Adhesion formation was more prominent in Groups IB, IIA, and IIB on day four only; abscesses were noted in the ischemic group in four rats. Anastomotic bursting pressure was significantly lower in sealed (IB) and ischemic anastomoses (IIA) than in normal anastomoses (IA) on day four. Sealing of ischemic anastomoses did not change bursting pressures on days two, four, and seven. The relative decrease of collagen in the sealed anastomoses is significantly higher on day four only. It is concluded that sealing of normal colonic anastomoses in the rat has a negative effect on wound healing. Ischemia at the anastomotic site results in weaker anastomotic strength on day four postoperatively. Also in ischemic anastomoses, fibrin sealant does not improve wound healing during the first seven days. Adhesion formation on ischemic intestinal anastomoses was not prevented by fibrin sealing.

Anastomosis, Surgical↗

Selective accumulation of endogenously produced porphyrins in a liver metastasis model in rats.

The possibility of using the porphyrin precursor 5-aminolevulinic acid to cause selective porphyrin accumulation in tumors was examined. Syngeneic colon carcinomas CC531 were implanted in the livers of Wag/Rij rats. Groups of three to six animals each were given 2 mg/mL of 5-aminolevulinic acid in drinking water from the 8th, 14th, or 17th day after tumor implantation. Two other groups received either 2.5 or 5 mg/kg of Photofrin II (Photomedica Inc., Raritan, NJ) intravenously on day 17. On day 19 the livers were removed and porphyrin concentrations were measured in normal livers and tumors by solvent extraction and high-performance liquid chromatography. Protoporphyrin accumulated progressively in tumors with increasing duration of 5-aminolevulinic acid administration (P = 0.0001), whereas no increase was found in normal livers. After 11 days of 5-aminolevulinic acid administration the porphyrin concentration ratio between tumors and livers was 4:1. In contrast, after Photofrin II administration the concentration was higher in normal livers than in tumors (1:3 ratio, tumor to liver). Enzyme measurements showed a threefold decrease in ferrochelatase activity in tumors compared with livers (P less than 0.001). In conclusion, oral administration of 5-aminolevulinic acid results in progressive accumulation of protoporphyrin in a transplantable colon carcinoma without accumulation in the surrounding liver tissue. This selective accumulation of porphyrins appears to be caused by a relative ferrochelatase deficiency in malignant tissue. 5-Aminolevulinic acid administration may be a suitable approach to photosensitizing liver tumors for photodynamic therapy or to early detection of tumors by fluorescence in ultraviolet light.

Aminolevulinic Acid↗

Treatment of experimental liver metastases with a noncontact neodymium: YAG laser.

The extent of tumor and liver damage after treatment with the neodymium:yttrium-aluminum-garnet (Nd:YAG) laser was investigated in a rat tumor model. Colon carcinoma CC531 was implanted in the liver and treated with 60 J (14 tumors), 120 J (15 tumors), or 180 J (12 tumors) at a power setting of 20 W. To assess the effects upon the tissues three animals were sacrificed immediately after treatment and 1, 2, 4, and 8 days later. Sections from the tumor sites were evaluated by light microscopy and the maximal depth and width of different zones of tissue damage were measured. Laser effects could be determined most accurately on Days 1 and 2 after treatment. Multiple linear regression analysis of the data indicated a linear relationship between laser energy and depth of tumor damage (P less than 0.01). The results of this study show the potential of the Nd:YAG laser to produce tumor coagulation necrosis with minimal liver necrosis.

Aluminum↗

Eicosanoids in breast cancer patients before and after mastectomy.

In 19 patients with a malignant breast tumor, tumor tissue and blood were taken to determine the eicosanoid profile and platelet aggregation. Values were compared with those of patients with benign tumors (n = 4), or undergoing a mammary reduction (n = 7). Postoperatively, blood was taken as well in order to compare pre- and postoperative values. Eicosanoids were measured in peripheral blood monocytes and mammary tissue by means of HPLC; furthermore, TXA2, 6-keto-PGF1 alpha, and PGE2 were determined by RIA. Differences in pre- and postoperative values of cancer patients were seen in plasma RIA values: PGE2 and 6-k-PGF1 alpha were significantly higher preoperatively when compared with postoperatively, however, such differences were seen in the control groups as well. Compared to benign tumor or mammary reduction test material the eicosanoid profile of tissue obtained from malignant mammary tumors showed important differences. Except for PGF2 alpha, HHT and 15-HETE no detectable quantities of eicosanoids were found in the non-tumor material, whereas in the malignant tumor material substantial quantities of a number of eicosanoid metabolites were present. Statistically significant correlations could be established between patient/histopathology data and the results of the platelet aggregation assays, e.g. between menopausal status and ADP aggregation; oestrogen receptor (+/-) and collagen and arachidonic acid aggregation, inflammatory cell infiltration score and arachidonic acid aggregation and fibrosis score and ADP aggregation. The results show that eicosanoid synthesis in material from mammary cancer patients is different from that in benign mammary tissue. The implications, in particular, in relation to future prognosis of the patient, remain obscure.

Breast Neoplasms↗

Interstitial photodynamic therapy in a rat liver metastasis model.

Photodynamic therapy (PDT) of hepatic tumours has been restricted owing to the preferential retention of photosensitizers in liver tissue. We therefore investigated interstitial tumour illumination as a means of selective PDT. A piece of colon carcinoma CC531 was implanted in the liver of Wag/Rij rats. Photofrin was administered (5 mg kg-1 i.v.) 2 days before laser illumination. Tumours with a mean (+/- s.e.) diameter of 5.7 +/- 0.1 mm (n = 106, 20 days after implantation) were illuminated with 625 nm light, at 200 mW cm-1 from a 0.5 cm cylindrical diffuser and either 100, 200, 400, 800 or 1600 J cm-1. Control groups received either laser illumination only, Photofrin only or diffuser insertion only. Short-term effects were studied on the second day after illumination by light microscopy and computer-assisted integration of the circumference of damaged areas. Long-term effects were studied on day 36. To determine the biochemistry of liver damage and function, serum ASAT and ALAT levels were measured on day 1 and 2, and antipyrine clearance on day 1. Tumour and surrounding liver necrosis increased with light dose delivered (P < 0.001). Best long-term results were obtained at 800 J cm-1 with complete tumour remission in 4 out of 6 animals. No deterioration in liver function was found. The results of this study show the ability of interstitial PDT to cause major destruction of tumour tissue in the liver combined with minimal liver damage.

Adenocarcinoma↗