[Erythema multiforme following COVID-19 vaccination (BNT162b2)]. / Erythema exsudativum multiforme infolge einer COVID-19-Impfung (BNT162b2).

We report a case of a patient with erythema multiforme major following COVID-19 (coronavirus disease 2019) vaccination. Lesions on skin and mucous membranes developed 48 h after the second dose of the mRNA-vaccine BNT162b2 (Tozinameran, Comirnaty®). Under the application of external glucocorticoids complete resolution was achieved within 3 weeks.

Risk Factors and Innovations in Risk Assessment for Melanoma, Basal Cell Carcinoma, and Squamous Cell Carcinoma.

Skin cancer is the most frequently diagnosed cancer globally and is preventable. Various risk factors contribute to different types of skin cancer, including melanoma, basal cell carcinoma, and squamous cell carcinoma. These risk factors encompass both extrinsic, such as UV exposure and behavioral components, and intrinsic factors, especially involving genetic predisposition. However, the specific risk factors vary among the skin cancer types, highlighting the importance of precise knowledge to facilitate appropriate early diagnosis and treatment for at-risk individuals. Better understanding of the individual risk factors has led to the development of risk scores, allowing the identification of individuals at particularly high risk. These advances contribute to improved prevention strategies, emphasizing the commitment to mitigating the impact of skin cancer.

Aluminium phthalocyanines-induced photolysis of human vascular wall cells in culture and the effect of fluoride on photodynamic action.

BACKGROUND: Phthalocyanines, second-generation photosensitizers with several attractive properties for use in photodynamic therapy, have been shown to accumulate in malignant lesions and atherosclerotic plaques. After exposure of phthalocyanines-loaded tissues to visible light, the targeted cells become injured and eventually die. In vitro, when fluoride is added before exposure to the light, it can protect some cell types against photodynamic action sensitized by chloroaluminium phthalocyanine (AIPc) and its derivatives. METHODS: The effect of 50 mumol/l chloroaluminium phthalocyanine tetrasulfonate (AIPcS4) and 5 mumol/l AIPc, with and without the addition of 5 mumol/l NaF, on the viability of cultured human endothelial cells (HuEC), human smooth muscle cells (HuSMC), and skin fibroblasts (HuSF) was examined. A 150 W quartz halogen light bulb equipped with a cut-off filter (lambda > 605 nm) was used to activate the phthalocyanines. Cell viability was examined 24 h after irradiation by staining of the cells with fluorescein diacetate/ethidiumbromide and by counting the number of attached cells with a cell counter. RESULTS: In the case of AIPc, the viability of all cell types tested was reduced, in a dose-dependent manner, to about 50% of that of the corresponding controls for a maximum irradiation time of 600 s. When we used AIPcS4, both HuEC and HuSF were considerably less sensitive than HuSMC. The addition of fluoride to AIPc-loaded cells before exposure to light protected HuEC and HuSF, but not HuSMC. In the case of AIPcS4, with and without fluoride, only HuSMC were sensitive. CONCLUSIONS: The addition of fluoride to AIPc or the use of AIPcS4 without fluoride could be a valuable approach, selectively destroying HuSMC without affecting HuEC and HuSF, for the reduction of restenosis rates after recanalization of stenosed or occluded arteries. Moreover, because neither type of phthalocyanines causes cutaneous phototoxicity, these second-generation photosensitizers seem to be best suited to the photodynamic treatment of atherosclerosis.

Expression of connective tissue growth factor (CTGF) mRNA in plaques of human anterior subcapsular cataracts and membranes of posterior capsule opacification.

PURPOSE: To investigate the correlation between connective tissue growth factor (CTGF) mRNA expression and immunohistochemical characteristics of anterior subcapsular cataract (ASC) formation as well as posterior capsule opacification (PCO) development (expression of type I collagen, alpha-smooth muscle actin and tenascin) under in vivo and under in vitro conditions in human and porcine lens epithelial cells. METHODS: CTGF mRNA expression was investigated using in situ hybridization and RT-PCR. Expression of type I collagen, alpha-smooth muscle actin and tenascin was detected by immunohistochemical staining. RESULTS: CTGF mRNA was expressed in human cataractous plaques of ASC and human PCO membranes, and appeared simultanously with the expression of type I collagen, alpha-smooth muscle actin and tenascin. CONCLUSION: The predominant expression of CTGF mRNA in human ASC and human PCO membranes suggests a significant role of CTGF in the pathological course of these ocular disorders.

[Effect of platelet-derived growth factor PDGF on replication of cultivated bovine lens epithelial cells]. / Einfluss des Platelet-derived-growth-Faktors PDGF auf replikative Vorgänge bei kultivierten bovinen Linsenepithelzellen (BLEZ).

It has previously been reported that PDGF isoforms AB and BB induce an increase of cytosolic free calcium in cultured bovine lens epithelial cells in a dose-dependent manner. To evaluate the biological capacity of PDGF, we investigated the proliferative response of bovine lens epithelial cells to stimulation with PDGF-AA, -AB and -BB. Since various unspecific components of serum-containing media act as mitogenes and mask the effect of PDGF, serum-free culture conditions were a prerequisite for growth-factor-induced effects. Therefore, a basic medium (Waymouth's MB 752/1 with Ham F12 Nutrient Mixture 1:2, v/v; Gibco BRL) was supplemented with only 2 mM CaCl2, 10 micrograms/ml Transferrin, 10 micrograms/ml Thyroglobulin (both Sigma Chemie) and standard amounts of antibiotics. PDGF isoforms were obtained by separate expression of cloned genes in Escherichia coli, which has been previously described. Under these conditions the isoforms PDGF-AB and -BB caused an increase in proliferation in a dose-dependent manner. The maximum increase of the cell number was 21% for PDGF-AB with an EC5 of 5 ng/ml. PDGF-BB revealed a maximum increase of 33% with an EC5 of 1.5 ng/ml. PDGF-AA, when used in similar concentration was ineffective. These data show the involvement of PDGF isoforms AB and BB in the replicative action of BLEC.

[Photodynamic activity of phthalocyanines in cultivated lens epithelial cells of the pig]. / Photodynamische Aktivität von Phthalozyaninen bei kultivierten Linsenepithelzellen vom Schwein.

One of the most frequent complications of extracapsular cataract surgery is posterior capsule opacification (PCO), which is caused by proliferation and migration of retained lens epithelial cells on the posterior capsule surface. This complication leads to a reduction in visual acuity. Depending on the age of the patients, the incidence varies between 15 and 50%. Selective elimination of these cells would therefore prevent PCO. Phthalocyanines (Pc) are efficient photosensitizers of various carcinoma cell lines. Their attractive property is the strong absorption of visible light above 600 nm. Upon absorption of a photon, the sensitizer in its excited state can be either directly cytotoxic or produce reactive oxygen species type I (free radicals) or type II (singlet oxygen), which become the mediator of cellular injury by affecting membranes and subcellular organelles. Preconfluent cultures of porcine lens epithelial cells were incubated with aluminum phthalocyanine (AlPc) or aluminum phthalocyanine transulfonate (AlPcS4) in concentrations ranging from 0-20 microM for 1 h at 37 degrees C. Thereafter, cultures were exposed to red light for 5 min. A 300 W quartz halogen light bulb equipped with a cut-off filter that allowed transmission of the whole spectrum above 600 nm was used for activation of the sensitizers. To evaluate the effect of fluoride, 5 mM NaF was added to the dye-loaded cells immediately before light exposure. This treatment had no toxic effect. Subsequently, the treated cells were incubated in culture medium for 24 h. After trypsinization, the cell number was determined by a cell counter. Prior to light exposure neither AlPc nor AlPcS4 was toxic in corresponding controls.(ABSTRACT TRUNCATED AT 250 WORDS)

[Serum-free cultivation of bovine stromal fibroblasts]. / Serumfreie Kultivierung boviner stromaler Fibroblasten.

UNLABELLED: The purpose of the present study was to conduct a comparative evaluation of the effect of several serum-free culture conditions on adhesion, population doubling, cryopreservation and PDGF-induced effects on cell proliferation of bovine stromal fibroblasts (BSF). Additionally, these effects were compared to serum-containing cultures. METHODS: Only second-passage BSF were used. Cells were cultured using four different culture media (WM/F12, WM/F12 + FCS 1%, LR-1, DMEM). After 24 h, plating efficiency was determined using a cell-counter system. Subsequently, the cells were seeded at a density of 100 cells/mm2 and cultured for 10 days using the different culture media. Cell number was determined at day 2, 4, 7 and 10 after seeding. Furthermore, the effect of 50 ng/ml PDGF-BB on the proliferation of BSF was tested for these conditions. Cell vitality was determined after cryopreservation of two weeks for each culture medium. RESULTS: The plating efficiency of BSF ranged from 50.2 to 55.5% for the serum-free culture media in contrast to serum-containing conditions, where plating efficiency was 94.8%. With WM/F12 + FCS 1%, a population doubling of 1.27 was observed after an incubation period of 10 days. In contrast, cultivation under serum-free conditions caused neither significant cell proliferation nor cell loss. The stimulation of cell proliferation with PDGF-BB was shown to be 28% (LR1), 40% (WM/F12 + FCS 1%) 76% (WM/F12) and 95% (DMEM) compared to the control. While cell vitality after cryo-preservation was found to be 62.7% using WM/F12 + FCS 1%, vitality using serum-free media was 12.6-22.8%. CONCLUSIONS: The results of the present study demonstrate that with respect to optimal cell adhesion and cell vitality after cryo-preservation, serum-containing media should be used. BSF cultured under the serum-free conditions used in the present study can be maintained quiescent and vital for at least 10 days. Therefore, these serum-free media are useful for cell-culture studies (e.g., determination of proliferation and cytotoxicity).

[Growth characteristics of bovine retinal pericytes in culture]. / Wachstumscharakteristika von bovinen retinalen Perizyten in Kultur.

The loss of retinal pericytes is one of the earliest changes in diabetic retinopathy. In order to study this phenomenon in vitro, an optimal isolation and cultivation system has to be established. Therefore, pericytes from bovine retinae were isolated enzymatically with 0.4% collagenase in phosphate-buffered saline and identical immunologically by positive staining with antibodies against smooth muscle alpha-actin. Routine cultivation of pericytes was performed by using DMEM supplemented with 10% fetal calf serum. Dependent on the in vitro age of cells, the effect of the following reagents on proliferative activity was determined: fetal calf serum, heparin, ECGF, ECGF+heparin, and glucose. Increasing serum concentrations stimulated the proliferation of pericytes, although the degree of stimulation was reduced with increasing in vitro age. Heparin inhibited the growth in a dose-dependent manner; the achieving 50% inhibition was extrapolated to be 25 micrograms/ml. ECGF increased pericyte proliferation significantly, with a maximum at 10 microliters/ml. In addition, ECGF reversed the inhibitory effect of heparin. Furthermore, all tested glucose concentrations (5.5-27.75 mmol/l) did not show any influence on growth rates of pericytes. The results demonstrate that routine cultivation of retinal pericytes is possible. Moreover, they indicate that enhanced blood glucose concentrations, as observed in diabetic patients, are not the only important factor in the loss of retinal pericytes.

[Effect of heparin and ascorbic acid on growth behavior of cultivated corneal epithelial cells of the rabbit]. / Wirkung von Heparin und Ascorbinsäure auf das Wachstumsverhalten kultivierter Hornhautepithelzellen des Kaninchens.

Although ascorbic acid and heparin are used for local therapy of corneal wounds that heal poorly (e.g., after chemical burns), little has been known up to now about the mechanisms underlying their effectiveness at the cellular level. The aim of the present study was therefore to evaluate the effect of heparin and ascorbic acid on the growth behaviour of corneal cells in vitro. For this purpose cell cultures from a corneal epithelial cell line were used. Stimulation of the cells with heparin at concentrations ranging from 10 to 200 micrograms/ml for 6 days led to a dose-dependent rise in growth rate (population doublings per day) of 0.48 +/- 0.50 to 2.19 +/- 1.65 (mean value +/- standard deviation, n = 10) with an EC50 of 132 micrograms/ml. In contrast, the addition of ascorbic acid at concentrations ranging from 0.5 to 3.0 mM led on average to a 40% dose-dependent inhibition of cell proliferation after 6 days, with an IC50 of 0.7 mM as-corbic acid. On the basis of these results, the use of heparin at concentrations of 180-200 micrograms/ml appears advantageous. In contrast, the local application of ascorbic acid for chemical burns with no stromal involvement should be subjected to a critical reassessment.

[Effect of lymphocyte-conditioned medium on expression of smooth muscle alpha-actin in lens epithelium cells in situ]. / Einfluss eines Lymphozytenkonditionierten Mediums auf die Expression des glattmuskulären alpha-Aktins in Linsenepithelzellen in situ.

PURPOSE: Contraction of the capsule of the ocular lens is based upon proliferation and contraction of transformed lens epithelial cells. It is assumed that these processes can be assisted by postoperative intraocular inflammation. Previously, we reported that lens epithelial cell proliferation is enhanced by lymphocyte-conditioned medium (LCM). In this study we investigated the effect of LCM as well as of a culture medium conditioned by pigmented ciliary epitheilal cells (CBCM) on the expression of the smooth-muscle alpha-actin of the contractile cytoskeletal elements. METHODS: Explants of the anterior lens capsule of freshly enucleated bovine eyes were cultured in serum-free LCM and CBCM for 3 days, followed by fixation. Smooth-muscle alpha-actin was identified by indirect immunoflorescence. Explants cultured in serum-free bFGF-containing and TGF-beta containing medium served as control. RESULTS: Lens epithelial cells expressed smooth-muscle alpha-actin under the influence of LCM or TGF-beta. No smooth muscle alpha-actin could be detected under the influence of CBCM or bFGF. CONCLUSION: Our results demonstrate that secreted molecules of activated lymphocytes are able to induce the transformation of lens epithelial cells into contractile myofibroblasts and may be involved in the post-operative contraction of lens capsules.

[PDGF-induced effect on cytosolic free calcium concentration of cultured retinal pericytes]. / PDGF-induzierte Wirkung auf die zytosolisch freie Kalziumkonzentration kultivierter retinaler Perizyten.

Although the selective loss of retinal pericytes has long been known to be one of the earliest histopathological findings in diabetic retinopathy, only limited information is available concerning their function and cell biology. Recently, it has been shown that the interaction of endothelial cells and pericytes plays an important role in the maintenance of vascular integrity. Additionally, it has been suggested that pericytes have a contractile function. Platelet-derived growth factor (PDGF), released from endothelial cells, has been shown to be a potent mitogen and vasoconstrictor. Cytosolic free calcium ([Ca2+]i) has been shown to play a key role as a second messenger for PDGF, involved in the regulation of various cellular functions, e.g. cell proliferation and vascular contractility. In order to characterize the effect of different PDGF homodimers on cultured bovine retinal pericytes, we investigated PDGF-AA- and -BB-dependent alterations in [Ca2+]i was determined with the Ca(2+)-sensitive fluorescent probe Quin-2. Basal levels were 118 +/- 30 nM. Stimulation with PDGF-BB in concentrations ranging from 5 to 20 ng/ml led to a dose-dependent increase of [Ca2+]i with an EC50 of 5.8 ng/ml. Maximum stimulation, to about 280% of basal levels, occurred after 3-4 min. In contrast, PDGF-AA was not effective. The results suggest that PDGF-BB may influence the integrity and contractility of the retinal microvasculature via modulation of the intracellular calcium homeostasis of pericytes. Additionally, it can be speculated that cultured retinal pericytes express mainly PDGF-beta-type receptors.

[Expression of platelet derived growth factor receptors in cultivated lens epithelial cells]. / Untersuchungen zur Expression von Platelet-derived-growth-Faktor (PDGF)-Rezeptoren an kultivierten Linsenepithelzellen.

It has been shown that PDGF plays a role in the regulation of lens growth and differentiation. PDGF occurs in vivo as a homodimer or heterodimer of the two polypeptide chains A and B. These isoforms bind with different affinities to two distinct receptor types, termed alpha and beta. In order to identify the different PDGF receptors on cultured bovine lens epithelial cells (BLEC), we performed receptor binding studies, using 125I-labelled PDGF isoforms. Analysis of our data revealed that BLEC expressed approximately 35,000 PDGF-BB binding sites (KD = 21 ng/ml) and 9000 PDGF-AB binding sites (KD = 11 ng/ml), but only 4800 PDGF-AA binding sites (KD = 7 ng/ml). This study represents the first demonstration that lens epithelial cells express PDGF receptors.

[Ascorbic acid. Cytotoxic effect on cultivated bovine lens epithelium cells]. / Die Askorbinsäure. Zytotoxischer Effekt auf kultivierte bovine Linsenepithelzellen.

Ascorbic acid is one of the main components (1.16 mM/l) of the aqueous humor. The molarity of this molecule is 25 times higher than in the plasma of the cow, man or horse. Now the question arises as to which function ascorbic acid has in this extremely high concentration referring to the proliferation of the lens epithelial cells. Thus, the effect of ascorbic acid was investigated upon bovine lens epithelial cells (BLEC) in the range of 0-3 mM/l. These cells were cultivated under various culture conditions (serum-free, serum-containing, aqueous-humor-containing medium) and also incubated with such mitogens as retinal extract (RE), crude endothelial cell growth factor (cECGF), basic fibroblast growth factor (bFGF) or with calcium. In each culture condition 1 mM/l ascorbic acid caused remarkable inhibition of the proliferation of BLEC. Higher concentrations (> 1.5 mM/l) revealed cytotoxic effects. These effects were independent of small variations in the pH value caused by ascorbic acid. In addition, the effect of 2 mM/l ascorbic acid in combination with catalase in a concentration of 500 Um/ml and 1000 Um/ml, respectively, was investigated. It could be shown that catalase is capable of preventing the cytotoxic effect of ascorbic acid. These results show the inhibitory effect of ascorbic acid in its physiological concentration in the proliferation of BLEC.

[Effect of heparin on proliferation of cultivated bovine lens epithelial cells]. / Wirkung von Heparin auf die Proliferation kultivierter boviner Linsenepithelzellen.

The treatments proposed to date for the prevention of secondary cataract have shown limited efficacy or have not been satisfactory due to ocular toxicity. Since it has been demonstrated that heparin can inhibit the proliferative activity of smooth muscle cells and fibroblasts in vitro and in vivo, we examined the effect of heparin at concentrations ranging from 20 to 200 micrograms/ml on the proliferation of cultured bovine lens epithelial cells (BLEC) under various culture conditions: (1) serum-free medium (SFM); (2) SFM + aqueous humor 1:1; (3) SFM +1 and 10% fetal calf serum; (4) SFM +1% retinal extract; (5) SFM +50 micrograms/ml endothelial cell growth factor; (6) SFM +10 ng/ml epidermal growth factor; (7) SFM +10 ng/ml basic fibroblast growth factor. Heparin caused no cytotoxic effects in any of the experiments. With medium 2 and 3, heparin caused dose-dependent inhibition of cell proliferation at concentrations ranging from 10 to 50 micrograms/ml. Cells cultivated in medium 4-7 with the addition of 50 micrograms/ml heparin revealed increased proliferative activity when compared with the corresponding controls. The antiproliferative activity on BLEC in medium containing aqueous humor suggests that heparin is a valuable tool for the prevention of secondary cataract in vivo.

A novel handheld fluorescent microarray reader for point-of-care diagnostic.

A novel handheld optical sensor for quantification of fluorescent microarrays, the so-called portMD-113 has been developed. On the surface of a planar waveguide, the spots of different fluorescently labeled biological complexes are excited by the evanescent field of the guided light. The emitted fluorescence signals of the spots are independently and simultaneously detected applying our system, which consists of a pinehole array, a microlens array, an interference filter and a detector array. As it is demonstrated in comparative measurements, the detection limit of this sensor is close to that of commercial top microarray readers, e.g. of modern laser scanners, while it has remarkable and important advantages over them. Namely, the device comprises only a few low-cost, lightweight and small components without applying any moving or energy-intensive elements, which results in turn in a commercially competitive, handheld and compact design and in the possibility to be supplied simply by a battery or a personal computer. These advantageous properties open prospects e.g. for point-of-care medical checks, as well.

A 2.5-Mb contig constructed from Angus, Longhorn and horned Hereford DNA spanning the polled interval on bovine chromosome 1.

The polled locus has been mapped by genetic linkage analysis to the proximal region of bovine chromosome 1. As an intermediate step in our efforts to identify the polled locus and the underlying causative mutation for the polled phenotype, we have constructed a BAC-based physical map of the interval containing the polled locus. Clones containing genes and markers in the critical interval were isolated from the TAMBT (constructed from Angus and Longhorn genomic DNA) and CHORI-240 (constructed from horned Hereford genomic DNA) BAC libraries and ordered based on fingerprinting and the presence or absence of 80 STS markers. A single contig spanning 2.5 Mb was assembled. Comparison of the physical order of STSs to the corresponding region of human chromosome 21 revealed the same order of genes within the polled critical interval. This contig of overlapping BAC clones from horned and polled breeds is a useful resource for SNP discovery and characterization of positional candidate genes.

An enhanced expression of ABC 1 transporter in circulating leukocytes as a potential molecular marker for the diagnostics of glaucoma.

OBJECTIVE: Glaucoma is a neurodegenerative disease. Since vascular dysregulation is supposed to be a risk factor for the development of glaucomatous damage, the preventive treatment might slow down the disease development. The efficiency of the therapeutic treatment depends particularly on a drug efflux pump regulated by ABC transporters. ABC 1 is also known to participate on the vascular regulation. This study was focused on the comparative analysis of ABC 1 expression levels in circulating leukocytes of non-glaucomatous individuals and glaucoma patients. RESULTS AND CONCLUSIONS: The expression rates of ABC 1 were significantly increased in leukocytes of glaucoma patients compared to non-glaucomatous individuals. The expression level of ABC 1 was, furthermore, highly homogeneous in glaucoma patients. In contrast, these expression levels in non-glaucomatous individuals were extremely heterogeneous. This transporter acts as the energy-dependent unidirectional transmembrane cholesterol efflux pump and can export a wide range of hydrophobic drugs. Additionally an observed enhanced ABC 1 expression in circulating leukocytes may be implicated in the vascular regulation mechanisms of glaucoma. We proposed the enhanced expression of ABC 1 in leukocytes as a potential marker for the diagnostics and ex vivo molecular monitoring of glaucoma.

Increased DNA breaks and up-regulation of both G(1) and G(2) checkpoint genes p21(WAF1/CIP1) and 14-3-3 sigma in circulating leukocytes of glaucoma patients and vasospastic individuals.

OBJECTIVE: Vascular disorder leading to local ischemia/reperfusion has been shown to play an important role in the glaucomatous damage. A decreased expression level of XPGC-gene has been found in circulating leukocytes of normal-tension glaucoma patients. Although decreased activity of XPGC-gene leads to insufficient DNA-repair, no leukopenia has been observed in glaucoma. Molecular mechanisms ensuring cell survival have not been elucidated yet for glaucoma with vascular disorder. MATERIAL AND METHODS: Using the ex vivo optical imaging method of alkaline "comet assay" comparative quantification of DNA breaks was performed in circulating leukocytes of non-glaucomatous non-vasospastic and vasospastic individuals as well as both normal-tension and high-tension glaucoma patients. Relative expression levels of the anti-apoptotic factors P21(WAF1/CIP1) and 14-3-3 sigma were investigated in all groups tested. RESULTS AND CONCLUSIONS: The quantification of P21(WAF1/CIP1) showed the highest expression rates in high-tension glaucoma patients which were significantly higher than those in all other groups tested. The highest expression rates of 14-3-3 sigma were found in both groups of glaucoma patients. These expression levels correlated well with DNA breaks measured. Since the expression of P21(WAF1/CIP1) in leukocytes was shown to be crucial for their survival under stress conditions, we suppose further that the up-regulation of this gene is the key event in the survival mechanisms of leukocytes in glaucoma accompanied with vascular disorder. The p21(WAF1/CIP1) gene should be further taken into consideration as a potential marker, the up-regulation of which in circulating leukocytes of vasospastic individuals may indicate an increased risk for the developing glaucoma.