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Biomedical subjects

J E Keunen

Publications and source records attributed to J E Keunen.

At least 19 recordsLinked to original sources

Short communication: Effects of subacute ruminal acidosis on free-choice intake of sodium bicarbonate in lactating dairy cows.

The effect of inducing subacute ruminal acidosis (SARA) on the free-choice intake of sodium bicarbonate (SB) was investigated in four midlactation Holstein cows in a switchover experiment with four 1-wk periods. The SARA was induced by replacing 25% of the ad libitum intake of total mixed ration (TMR) with pellets containing 50% ground wheat and 50% ground barley and restricting access to TMR from 0700 to 1700 h. Control consisted of feeding TMR ad libitum. Powdered SB was provided for ad libitum consumption. Rumen pH was measured continuously using indwelling pH probes. Induction of SARA reduced (P < 0.05) the average daily rumen pH from 6.08 to 5.87, increased (P < 0.05) the average duration of rumen pH below 6 from 547 min x d(-1) to 916 min x d(-1), and increased (P < 0.05) the average duration of rumen pH below 5.6 from 132 min x d(-1) to 397 min x d(-1) (P < 0.05) but did not significantly affect SB intake. Average intake of SB was 26.8 g x d(-1) during SARA and 34.5 g x d(-1) during control. These low SB intakes must not have substantially affected rumen pH. Sodium bicarbonate intake differed significantly (P < 0.05) between cows. These data indicate that cows did not select SB in order to attenuate SARA.

Acidosis↗

Effects of a subacute ruminal acidosis model on the diet selection of dairy cows.

Two experiments were conducted to study the effects of a subacute ruminal acidosis (SARA) model on diet choice in dairy cows. In the first experiment, 25% of the ad libitum dry matter intake (DMI) of the total mixed ration (TMR) was replaced with wheat-barley pellets (WBP, 50% ground wheat, 50% ground barley). Rumen pH was measured continuously via in-dwelling probes in 4 mid to late lactation cows. This diet change reduced rumen pH by 0.14 +/- 0.02 pH units (mean +/- SE) and increased time below pH 6.0, from 319 +/- 36 min(-1) to 641 +/- 36 min(-1). Hence, the nutritional model successfully induced SARA. The second experiment determined if inducing SARA increases the feed preference for long alfalfa hay compared with alfalfa pellets. The 2 wk of inducing SARA were separated by 1 control wk. Four cows on either SARA and control diets were given a choice of 2 feeds, 2 times per d, for 30 min. The preference ratios (PR = Amount of Hay consumed/Amount of Hay + Pellets consumed) for alfalfa hay during two SARA weeks was greater (0.85 +/- 0.03) compared with the control week (0.60 +/- 0.03). In SARA weeks, average rumen pH was 0.23 +/- 0.03 units lower, and time below pH 6.0 and 5.6 was higher compared to control. These results suggest that when given a choice of feeds, dairy cows alter their diet selection to attempt to attenuate SARA.

Acidosis↗

Temperature dependence of thermal damage to the sclera: exploring the heat tolerance of the sclera for transscleral thermotherapy.

Thermal damage to the human sclera in relation to temperature and duration of exposure was studied in order to determine the heat tolerance of the sclera with respect to transscleral thermotherapy of choroidal melanoma. Samples of human sclera were submerged in saline for 10 sec to 10 min at temperatures of 37-100 degrees C. The effects of heat on the shape, weight and size of the samples were studied. Thermal damage of scleral collagen was examined by polarized light microscopy (LM) and electron microscopy (EM). The sclera was embedded in epoxy resin and stained with toluidine blue for LM and with uranyl acetate and lead citrate for EM. Thermal damage of scleral collagen on polarized LM was graded on a five point scale. Scleral damage was visible on macroscopic examination and on LM and EM in sclera heated at 65 degrees C for 20 sec or longer, at 70 degrees C for 10 sec or longer, and at higher temperatures. A sigmoidal function was used to define the relation between temperature and changes in diameter, thickness, and weight of scleral samples. Using fitted functions, the threshold temperature for thermal damage was estimated to be 59-61 degrees C when samples were heated for 10 min, 62-63 degrees C when heated for 1 min, and 66-67 degrees C when heated for 10 sec; the threshold exposure time at 60 degrees C was estimated to be 7-12 min. These results indicate a temperature of 60 degrees C for 1 min is well tolerated by human donor sclera; information of in vivo studies is required to validate whether this setting can be used in transscleral thermotherapy (TSTT) for choroidal melanoma.

Adolescent↗

Additional cell damage after transpupillary thermotherapy in choroidal malignant melanoma.

The aim of this study was to evaluate the effectiveness of transpupillary thermotherapy (TTT) in generating tumour necrosis by light and electron microscopy, as well as to evaluate additional cell damage in the area directly adherent to the necrotic zone. Four eyes of four patients diagnosed with intraocular malignant melanoma of the uvea were treated experimentally with diode laser TTT. In all cases a standard technique was used. All eyes were enucleated: one eye the day after TTT, two eyes 2 days after TTT, and one eye 6 weeks after TTT. Immediately after enucleation the eyes were immersed in standard Karnovsky's fixative with cocodylate buffer and prepared for light and electron microscopy. In the treated area of all four melanomas we found a dense band of necrotic tissue (zone A) consisting of an amorphous mass of dead cells sharply demarcated from the rest of the neoplastic tissue. Next to this zone was a more eosinophilic and also sharply demarcated band (zone B) that consisted of similar but less intensive changes. In the next band (zone C), marked injury to the cellular membrane and subcellular structures were seen on electron microscopy. The next band (zone D) consisted of changes mainly observed only within the cytoplasm of neoplastic cells and significantly less intensive than those in zone C. Outside zone D tumour cells that were normal in appearance were seen. No scleral alterations induced by heat were found. We concluded that after TTT the cytotoxic effect gradually decreases in proportion to the distance from the central point of the diode laser spot, with additional cell damage in the area adjacent to the necrotic zone. The interval between TTT and enucleation had no influence on the histological results.

Cell Nucleus↗

Corneal autofluorescence in choroidal melanoma or in choroidal naevus.

AIMS: To investigate whether corneal autofluorescence is different in patients with choroidal melanoma or choroidal naevus. METHODS: Corneal autofluorescence was determined by fluorophotometry in both eyes of 32 patients with a unilateral choroidal melanoma, 32 patients with a unilateral choroidal naevus, and 32 age matched healthy controls. The corneal autofluorescence ratio between affected and contralateral eyes of patients or between randomly selected eyes of healthy controls was calculated. RESULTS: Mean corneal autofluorescence ratio of patients with a choroidal melanoma was significantly higher than that of healthy controls (mean ratio: 1.09 (SD 0.15) and 1.00 (0.09), respectively, ANOVA p=0.014), and than that of patients with choroidal naevus (mean ratio 0.96 (0.09), p<0.001). Mean ratios of patients with choroidal naevus and healthy controls were not significantly different (p=0.27). CONCLUSIONS: Corneal autofluorescence ratio of patients with a unilateral choroidal melanoma is increased. This is probably due to an increased flow of glucose through the impaired blood-aqueous barrier in the affected eye, resulting in additional glycation of corneal proteins and hence in increased autofluorescence. The corneal autofluorescence is not increased in patients with a choroidal naevus, because the blood-aqueous barrier is not impaired in the affected eye in these patients. Measurement of corneal autofluorescence is simple, fast, and non-invasive, and might be helpful to distinguish between patients with choroidal melanoma and those with choroidal naevus.

Analysis of Variance↗

Characterization of melanocortin-1 receptor gene variants in uveal melanoma patients.

PURPOSE: Allelic variations of the melanocortin-1 receptor (MC1R) gene have been linked to red hair and sun-sensitive skin types and may play a role in the susceptibility to develop cutaneous malignant melanoma (CMM). To define the role of MC1R gene in uveal melanoma, a case control study was performed, in which the presence of MC1R gene variations in uveal melanoma patients was compared with that of healthy controls. METHODS: MC1R gene variants were analyzed in 162 uveal melanoma patients and 255 healthy controls. After genomic DNA was isolated from venous blood, the MC1R gene was amplified by polymerase chain reaction (PCR) and examined for the presence of variants by single-strand conformation polymorphism (SSCP) analysis. Participants were asked to complete a questionnaire regarding skin type, eye color, and hair color. RESULTS: No disparity was found between the distribution of the MC1R gene variants in both groups. Furthermore, no associations between MC1R genotype and pigment phenotype were found. In contrast to CMM, uveal melanoma patients did not show specific MC1R gene variants. Compared with controls, most uveal melanoma patients had blue eyes (65%, P = 0.060) and skin type III (56%); however, in the uveal melanoma group the presence of dark blond hair was significantly elevated (46%, P = 0.030). These findings are in contrast with studies on CMM, where most patients have skin type II and red/fair hair. CONCLUSIONS: These data suggest that MC1R variants do not play a role in the susceptibility to develop uveal melanoma. Furthermore, most uveal melanoma patients share phenotypic characteristics that differ from findings in CMM patients.

Adult↗

Expression of epidermal growth factor receptor: risk factor in uveal melanoma.

PURPOSE: To investigate the prognostic significance of the expression of epidermal growth factor receptor (EGFR) in uveal melanoma. EGFR is a transmembrane glycoprotein, and its expression has been correlated with the development of metastases in various malignancies. METHODS: Frozen sections from 22 primary uveal melanomas were examined for EGFR expression by a three-step immunoperoxidase staining, using a mouse anti-human EGFR IgG2b monoclonal antibody. The results were compared with patient survival and clinical and histopathologic parameters. RESULTS: EGFR expression could not be determined on one tumor due to excessive pigmentation. Two patients died of causes unrelated to melanoma, and two patients were lost to follow-up. Out of 21 tumors, six tumors showed immunoreactivity for EGFR. Five of these six patients (83%) died due to metastases, compared with 2 (17%) of 12 patients with no EGFR expression (Kaplan-Meier analysis P = 0.0004). EGFR-positive tumors tended to have a greater tumor prominence and a higher mitotic rate. CONCLUSIONS: The expression of EGFR was significantly correlated with death due to metastatic disease and therefore can be regarded as an important prognostic factor in human uveal melanoma.

Adult↗

Effects of transpupillary thermotherapy on immunological parameters and apoptosis in a case of primary uveal melanoma.

Transpupillary thermotherapy (TTT) is a new treatment modality for uveal melanoma. We studied whether application of TTT influences the immunogenicity of the tumour cells in vivo or the expression of molecules related to apoptosis. Immunohistochemistry using monoclonal antibodies directed against HLA molecules, HMB45, P53, Fas ligand (FasL), Fas, Bcl-2 and tumour-infiltrating cells was applied to sections of an enucleated eye containing a uveal melanoma that received TTT 1 week before enucleation. The innermost part of the tumour which had been exposed directly to the laser treatment showed no staining for HLA antigens, nor for Fas or FasL epitopes. The intermediate part of the tumour showed a wet necrosis and HLA expression similar to the expression in the peripheral tumour. A large number of macrophages were observed in the necrotic as well as the intact tumour tissue, especially bordering the wet necrotic area. FasL and Bcl-2 were only expressed in the viable, outer part of the tumour. This immunological evaluation of one case of uveal melanoma treated with TTT revealed that TTT may not only have a direct destructive effect on the primary tumour, but may also influence the immunogenicity of uveal melanoma cells, induce infiltration of macrophages into the tumour, and induce apoptosis. The presence of many macrophages suggests that they play a role in the removal of the TTT-treated tumour tissue by phagocytosis.

Adult↗

[The laser pointer: no demonstrated danger to the eyes].

If laser pointers are powerful enough (> 5 mW), they can cause ocular damage. Most laser pointers in use, however, have low power, viz. 1 mW. In the peer-reviewed scientific literature worldwide not a single case of eye damage due to laser pointers is described. A review among Dutch ophthalmologists up to June 1998 revealed no cases of permanent damage caused by laser pointers. In view of the widespread use of laser pointers, the risk of retinal damage must be minimal, even with the types now banned. Laser pointers of 1 mW emitting light red or green light have sufficient visibility on projection screens. It is advisable to prohibit the sale of more powerful pointers to prevent excesses.

Eye Injuries↗

Transpupillary thermotherapy: results in 50 patients with choroidal melanoma.

OBJECTIVE: To evaluate the efficacy and safety of transpupillary thermotherapy in treating choroidal melanoma. METHODS AND PATIENTS: To perform transpupillary thermotherapy, infrared diode laser energy at 810 nm was used with a beam diameter of about 3 mm and 1-minute exposure time. All 50 patients had choroidal melanoma. We performed transpupillary thermotherapy in 21 tumors that had responded insufficiently to 800 Gy ruthenium 106 brachytherapy; it was combined with 800 Gy106Ru brachytherapy for 10 tumors greater than 5 mm in height and with 600 Gy for 19 tumors 5 mm or less in height. RESULTS: All but 1 tumor exhibited reduction in tumor height within a mean follow-up of 20.5 months (range, 6-49 months). In 41 eyes (82%), the tumor flattened completely. Visual acuity was 20/60 or better in 43 eyes (86%) before treatment and in 14 eyes (28%) at the last examination because of radiation vasculopathy. Neovascular glaucoma developed in 1 eye, and total retinal detachment developed in 2 eyes. Tumor recurrence was observed in 1 patient. CONCLUSIONS: Although long-term results are necessary to properly appraise this new therapy, transpupillary thermotherapy may be useful as a complementary modality to brachytherapy.

Adult↗

Long term follow-up of lenticular autofluorescence and transmittance in healthy volunteers.

This study was undertaken to assess long term changes in the lenticular autofluorescence and transmittance of healthy volunteers. Both eyes of 15 healthy volunteers aged 8 to 62 years were examined by slit-lamp examination and fluorophotometric measurements of lenticular autofluorescence (lambdaexc=415 nm-490 nm, lambdaem= 510 nm-550 nm) and transmittance (lambda=415 nm-550 nm) in 1983 and 1996. The changes in lenticular autofluorescence and transmittance between 1983 and 1996 were determined and compared with those calculated on the basis of age-dependence curves of a group of 56 healthy volunteers measured in 1983. These curves were obtained by approximation of the values of the 56 healthy volunteers by a linear, polynomial or exponential function of age, respectively. A mean yearly increase of lenticular autofluorescence of 8.69 ng . equivalent fluorescein . ml-1 . year-1 was observed in the 15 volunteers between 1983 and 1996 and this increase was significantly larger than that calculated on the basis of the three age-dependence curves in 1983 (6.36, 6.47 and 6.53 ng . equivalent fluorescein . ml-1 . year-1, P=0.0026, 0.0037 and 0.0044, respectively). The transmittance showed a mean yearly decrease of 0.508% . year-1 which was significantly larger than calculated on the basis of the three age-dependence curves in 1983 (0.153, 0.220 and 0.183% . year-1, P=0. 0033, 0.012 and 0.0059, respectively). One volunteer, i.e. the individual with the highest lenticular autofluorescence and lowest transmittance in 1983, had developed cataract in 1996. These measurements demonstrate for the first time, quantitative changes in lenticular autofluorescence and transmittance in a long term follow up. The at least 35% larger increase of autofluorescence and 130% larger decrease in transmittance in the follow-up group between 1983 and 1996 in comparison with the values based on age-dependent curves in 1983 could be caused by the increased solar UV radiation in The Netherlands during that period of time.

Adolescent↗

Fruits and vegetables that are sources for lutein and zeaxanthin: the macular pigment in human eyes.

BACKGROUND: It has been suggested that eating green leafy vegetables, which are rich in lutein and zeaxanthin, may decrease the risk for age related macular degeneration. The goal of this study was to analyse various fruits and vegetables to establish which ones contain lutein and/or zeaxanthin and can serve as possible dietary supplements for these carotenoids. METHODS: Homogenates of 33 fruits and vegetables, two fruit juices, and egg yolk were used for extraction of the carotenoids with hexane. Measurement of the different carotenoids and their isomers was carried out by high performance liquid chromatography using a single column with an isocratic run, and a diode array detector. RESULTS: Egg yolk and maize (corn) contained the highest mole percentage (% of total) of lutein and zeaxanthin (more than 85% of the total carotenoids). Maize was the vegetable with the highest quantity of lutein (60% of total) and orange pepper was the vegetable with the highest amount of zeaxanthin (37% of total). Substantial amounts of lutein and zeaxanthin (30-50%) were also present in kiwi fruit, grapes, spinach, orange juice, zucchini (or vegetable marrow), and different kinds of squash. The results show that there are fruits and vegetables of various colours with a relatively high content of lutein and zeaxanthin. CONCLUSIONS: Most of the dark green leafy vegetables, previously recommended for a higher intake of lutein and zeaxanthin, have 15-47% of lutein, but a very low content (0-3%) of zeaxanthin. Our study shows that fruits and vegetables of various colours can be consumed to increase dietary intake of lutein and zeaxanthin.

Carotenoids↗

Lens transmission by fluorophotometry after brachytherapy and thermotherapy of choroidal melanoma.

We studied the effect of transpupillary thermotherapy (TTT) by a diode laser at 810 nm combined with episcleral ruthenium-106 plaque treatment (106Ru) on lens transparency in patients with choroidal melanoma. Lens transmission of blue-green light was measured by fluorophotometry in 17 patients treated with 106Ru treatment and TTT (measured 0.36 years after treatment), 12 patients treated with 106Ru alone (measured 19 years after treatment) and 25 age-matched healthy controls. Differences in lens transmission were not significant between treated and untreated fellow eyes (p > 0.15) nor between patient and control eyes (p > 0.25). TTT of choroidal melanoma combined with 106Ru plaque irradiation did not have a significant effect on the lens transparency up to 6 years after treatment.

Aged↗