Prognostic relevance of topoisomerase II α and minichromosome maintenance protein 6 expression in colorectal cancer.

BACKGROUND: Despite rising incidence rates of colorectal malignancies, only a few prognostic tools have been implemented in proven clinical routine. Cell division and proliferation play a significant role in malignancies. In terms of colorectal cancer, the impact of proliferation associated proteins is controversially debated. The aim of our study was to examine the expression of topoisomerase II α and minichromosome maintenance protein 6 and to correlate these findings with the clinical data. METHODS: Tissue samples of 619 patients in total were stained using the antibodies Ki-S4 and Ki-MCM6 targeting topoisomerase II α as well as minichromosome maintenance protein 6. The median rate of proliferation was correlated with clinical and follow up data. RESULTS: The expression rate of minichromosome maintenance protein 6 is significantly higher than the proportion of topoisomerase II α in tumour cells (p < 0.001). A high expression of both proteins coincides with a beneficial outcome for the patient, indicating a favourable prognostic marker (p < 0.001 and p = 0.008). CONCLUSIONS: We have demonstrated that high expression rates of proliferative markers is linked to a beneficial patient outcome. According to the general opinion, a high expression rate correlates with a poor patient outcome. In this study, we were able to refute this assertion.

Therapeutic potential of larval excretory/secretory proteins of the pig whipworm Trichuris suis in allergic disease.

BACKGROUND: Gastrointestinal nematodes are currently being evaluated as a novel therapeutic in the treatment of chronic human inflammatory disorders, due to their unique ability to induce immunoregulatory pathways in their hosts. In particular, administration of ova from the pig whipworm Trichuris suis (T. suis; TSO) has been proposed for the treatment of allergic, inflammatory and autoimmune disorders. Despite these advances, the biological pathways through which TSO therapy modulates the host immune system in the context of human disease remain undefined. METHODS: We characterized the dominant proteins present in the excretory/secretory (E/S) products of first-stage (L1) T. suis larvae (Ts E/S) using LC-MS/MS analysis and examined the immunosuppressive properties of whole larval Ts E/S in vitro and in a murine model of allergic airway disease. RESULTS: Administration of larval Ts E/S proteins in vivo during the allergen sensitization phase was sufficient to suppress airway hyperreactivity, bronchiolar inflammatory infiltrate and allergen-specific IgE production. Three proteins in larval Ts E/S were unambiguously identified. The immunomodulatory function of larval Ts E/S was found to be partially dependent on the immunoregulatory cytokine IL-10. CONCLUSIONS: Taken together, these data demonstrate that the released proteins of larval T. suis have significant immunomodulatory capacities and efficiently dampen allergic airway hyperreactivity. Thus, the therapeutic potential of defined larval E/S proteins should be exploited for the treatment of human allergic disorders.

Dietary fiber and its role in performance, welfare, and health of pigs.

Dietary fiber (DF) is receiving increasing attention, and its importance in pig nutrition is now acknowledged. Although DF for pigs was frowned upon for a long time because of reductions in energy intake and digestibility of other nutrients, it has become clear that feeding DF to pigs can affect their well-being and health. This review aims to summarize the state of knowledge of studies on DF in pigs, with an emphasis on the underlying mode of action, by considering research using DF in sows as well as suckling and weaned piglets, and fattening pigs. These studies indicate that DF can benefit the digestive tracts and the health of pigs, if certain conditions or restrictions are considered, such as concentration in the feed and fermentability. Besides the chemical composition and the impact on energy and nutrient digestibility, it is also necessary to evaluate the possible physical and physiologic effects on intestinal function and intestinal microbiota, to better understand the relation of DF to animal health and welfare. Future research should be designed to provide a better mechanistic understanding of the physiologic effects of DF in pigs.

Molecular cloning of an immunodominant antigen of Onchocerca volvulus.

We have cloned and characterized the gene for an immunodominant antigen of O. volvulus that is recognized by the sera of 96% of patients with onchocerciasis. Its 1.2-kb mRNA constitutes 0.3% of adult worm poly(A)+ RNA and its cDNA sequence reveals that it is not a highly conserved structural protein such as actin or tubulin. Similar but not identical genes occur in the genomes of related filarie, Brugia malayi and Dirofilaria immitis. The recombinant antigen has both immunodiagnostic and immunoprophylactic significance.

The angiotensin II type 2 (AT2) receptor promotes axonal regeneration in the optic nerve of adult rats.

The renin-angiotensin system (RAS) has been traditionally linked to blood pressure and volume regulation mediated through the angiotensin II (ANG II) type 1 (AT1) receptor. Here we report that ANG II via its ANG II type 2 (AT2) receptor promotes the axonal elongation of postnatal rat retinal explants (postnatal day 11) and dorsal root ganglia neurons in vitro, and, moreover, axonal regeneration of retinal ganglion cells after optic nerve crush in vivo. In retinal explants, ANG II (10(-7)-10(-5) M) induced neurite elongation via its AT2 receptor, since the effects were mimicked by the AT2 receptor agonist CGP 42112 (10(-5) M) and were entirely abolished by costimulation with the AT2 receptor antagonist PD 123177 (10(-5) M), but not by the AT1 receptor antagonist losartan (10(-5) M). To investigate whether ANG II is able to promote axonal regeneration in vivo, we performed optic nerve crush experiments in the adult rats. After ANG II treatment (0.6 nmol), an increased number of growth-associated protein (GAP)-43-positive fibers was detected and the regenerating fibers regularly crossed the lesion site (1.6 mm). Cotreatment with the AT2 receptor antagonist PD 123177 (6 nmol), but not with the AT1 receptor antagonist losartan (6 nmol), completely abolished the ANG II-induced axonal regeneration, providing for the first time direct evidence for receptor-specific neurotrophic action of ANG II in the central nervous system of adult mammals and revealing a hitherto unknown function of the RAS.

Characterization of an immunodominant Onchocerca volvulus antigen with patient sera and a monoclonal antibody.

Adult Onchocerca voluvlus and infective larvae, but not microfilariae contain an immunodominant antigen (33,000 and 21,000 Mr in females, 39,000, 33,000, and 21,000 Mr in males, 133,000 Mr in infective larvae) which is recognized by an Onchocerca-specific mAb. The component is part of the reproductive organs and muscles. 96.2% of onchocerciasis sera contained antibodies detectable by immunoblotting against it. Antigen purified by immunoaffinity chromatography was specifically recognized in immunoblots by onchocerciasis sera, but not by sera from other filarial infections. The high immunogenicity, the specificity, and the occurrence in infective larvae of this antigen indicate an immunodiagnostic potential and a possible role in the immunobiology of the parasite.

Gastrointestinal nematode infection interferes with experimental allergic airway inflammation but not atopic dermatitis.

BACKGROUND: Some helminth infections are negatively associated with the prevalence of allergic disorders, arguing for a modulation of allergic reactions by the parasites, depending on the worm species, intensity and phase of infection and the type of disease. OBJECTIVE: The aim of this study was to analyse the influence of a chronic infection with the gastrointestinal nematode Heligmosomoides polygyrus, in a murine model of allergic airway disease and of atopic dermatitis (AD), respectively. METHODS: Mice were infected with H. polygyrus and systemically sensitized with the model allergen ovalbumin. Subsequently, the animals were challenged with the allergen either via the airways for induction of airway disease, or via skin patches for induction of dermatitis. RESULTS: Mice concomitantly infected with H. polygyrus showed diminished eosinophil and lymphocyte recruitment into the lungs and decreased allergen-specific IgE levels when compared with sensitized and airway challenged controls. In addition, animals showed a trend towards reduced airway hyper-reactivity. In contrast, no significant differences in the severity of eczematous skin lesions were observed between infected and control animals in the AD model. Although H. polygyrus infection reduced CD8+ and CD4+ T-cell infiltration into the skin and production of allergen-specific IgE, mast cell recruitment was significantly increased in worm-infected mice in the dermatitis model. The worm infection was associated with significantly elevated numbers of Foxp3+ regulatory T cells (Treg) in peribronchial lymph nodes in H. polygyrus-infected sensitized and airway challenged mice. In contrast, Treg cells were basically absent in eczematous skin and their number was not increased in skin-draining lymph nodes of mice with experimental dermatitis. CONCLUSION: Infection with the gastrointestinal nematode used in our study leads to significant inhibition of mucosa-associated but not cutaneous allergic reactions, pointing to a site specificity of the immunomodulation exerted by helminths. This finding might be an important aspect for future considerations of helminths for treatment of allergic diseases.

Suppression of MAP kinases inhibits microglial activation and attenuates neuronal cell death induced by alpha-synuclein protofibrils.

Alpha-Synuclein (alpha-Syn) accounts, as a major component of Lewy bodies (LB), for the filamentous deposits in many cases of neurodegenerative diseases. Yet, little is known about the molecular mechanisms of neuronal loss in these diseases. The correlation between alpha-Syn oligomerization/aggregation and pathologies raises the key question of which molecular form of alpha-Syn (i.e. monomeric alpha-Syn, protofibrils or mature fibrils) represents the damage-inducing culprit in the scenario of synucleinopathies. We show that human alpha-Syn protofibrils (PFs) are potent activators of parallel proinflammatory signalling pathways (p38 and ERK1/2 MAP kinases and NF-kappaB) in microglial cells in vitro. Furthermore, stereotactic injection of alpha-Syn PFs into the substantia nigra of adult rats leads to a profound activation of microglia and adjacent neuronal cell loss, which can be attenuated by the MAP kinase inhibitor semapimod. We propose that the neurodegenerative process of alpha-synucleinopathies involves microglial activation through alpha-Syn released or extruded from cells with pathogenic alpha-Syn metabolism. Compounds that inhibit the MAPK/NF-kappaB pathways might be a promising pharmacological strategy for the treatment of the inflammatory component of synucleinopathies including PD.

Involvement of formyl-peptide-receptor-like-1 and phospholipase D in the internalization and signal transduction of amyloid beta 1-42 in glial cells.

Recent studies suggest that the formyl-peptide-receptor-like-1 (FPRL1) plays an essential role in the inflammatory responses of host defense mechanisms and neurodegenerative disorders such as Alzheimer's disease (AD). We therefore analyzed whether amyloid beta1-42 (Abeta1-42) increased the activity of phospholipase D (PLD) via FPRL1, which is an enzyme involved in the secretion, endocytosis and receptor signaling. PLD activity was determined using a transphosphatidylation assay. The internalization of Abeta1-42 via FPRL1 was visualized using fluorescence microscopy and quantified by ELISA (Enzyme Linked Immunosorbent Assay). Determining receptor activity by extracellular-signal regulated kinases 1/2 (ERK1/2) phosphorylation and cAMP level measurement verified the Abeta1-42-induced activation of FPRL1. We were able to show that Abeta1-42 is rapidly internalized via FPRL1 in astrocytes and microglia. PLD was additionally activated by Abeta1-42 and via FPRL1 in rat glial cells. Furthermore, the ERK1/2 phosphorylation by FPRL1 agonists was dependent on the PLD product phosphatidic acid (PA). Together, these data suggest that PLD plays an important role in the regulation of Abeta1-42-induced endocytosis and FPRL1 receptor signaling.

Vaccination of eels (Anguilla japonica and Anguilla anguilla) against Anguillicola crassus with irradiated L3.

The original host of the swimbladder nematode Anguillicola crassus, the Japanese eel (Anguilla japonica) and the recently colonized European eel (Anguilla anguilla) were immunized with 40 irradiated (500 Gy) 3rd-stage larvae (L3) of this parasite and challenged with an infection of 40 normal L3. The immunization induced a significant reduction of the number of adult worms developing from the challenge infection in A. japonica, but not in A. anguilla. The induced resistance (calculated using the relation of the number of adult worms in immunized eels and in non-immunized control eels) in A. japonica was 87.3%+/-30.4%. Following a single infection, the percentage of adult worms found in A. japonica was lower as compared to A. anguilla, and the few adult worms were much smaller, revealing a lower susceptibility of A. japonica to A. crassus in comparison to A. anguilla. Both eel species developed an antibody response against A. crassus, but the level of antibody responses was not positively correlated with the protection against infection, suggesting that the antibody response is not a key element in resistance of eels against A. crassus. This study suggests that the original host of A. crassus is able to mount efficient protective immune responses against its parasite, whereas the newly acquired host seems to lack this ability.

Characterisation of recombinant immunoreactive antigens of the scab mite Sarcoptes scabiei.

Sarcoptic mange (or scabies) is an important skin disease which can affect a variety of species including humans, cattle, goats, sheep, horses, pigs, rabbits, and dogs. Approximately 300 million people are affected worldwide and in lifestock animals the infestation may lead to substantial economic losses caused by depression in growth and feed conversion rates. Diagnosis of Sarcoptes infestation is difficult and only a few serological tests have been developed using whole mite antigen for diagnosis of mange in animals. Here we describe the isolation and characterisation of cDNAs of several immunoreactive clones and their recombinant expression in Escherichia coli. Three of the proteins contain repetitive sequences which suggests that they might be involved in immune evasion. The application of these antigens in serodiagnosis and the suitability for diagnosis is discussed.

Serological cross-reactivity between a human Ro/SS-A autoantigen (calreticulin) and the lambda Ral-1 antigen of Onchocerca volvulus.

We have cloned and sequenced a 46-kD Ro/SS-A autoantigen gene that is the human homologue of the calcium-binding protein, calreticulin. The sequence of this 46-kD Ro/SS-A protein (calreticulin) has significant homology to lambda Ral-1, a recombinant cDNA clone corresponding to a major antigen of the nematode, Onchocerca volvulus, the infectious agent of onchocerciasis. We therefore sought to determine whether antibodies produced by onchocerciasis patients might crossreact with the human 46-kD Ro/SS-A autoantigen (calreticulin). 20 of 22 sera from Liberian onchocerciasis patients who had no known evidence of autoimmune disease were found to contain antibodies that reacted with the 46-kD Ro/SS-A (calreticulin) by immunoblot analysis. Characteristic of sera reactive with Ro/SS-A antigens, some onchocerciasis sera also immunoprecipitated the Ro/SS-A-associated hY RNAs. In addition, a monoclonal antibody raised against O. volvulus organisms reacted to purified human WiL-2 cell 46 kD Ro/SS-A antigen (calreticulin) by ELISA. These results strongly suggest that onchocerciasis patients produce antibodies that crossreact with the 46-kD human Ro/SS-A autoantigen (calreticulin) and raise the possibility that infectious organisms such as O. volvulus might play a triggering or exacerbating role in the human Ro/SS-A autoimmune response.

Tetracycline therapy targets intracellular bacteria in the filarial nematode Litomosoides sigmodontis and results in filarial infertility.

Intracellular bacteria have been described in several species of filarial nematodes, but their relationships with, and effects on, their nematode hosts have not previously been elucidated. In this study, intracellular bacteria were observed in tissues of the rodent parasite Litomosoides sigmodontis by transmission electron microscopy and by immunohistochemistry using antiendobacterial heat shock protein-60 antisera. Molecular phylogenetic analysis of the bacterial 16S ribosomal RNA gene, isolated by PCR, showed a close relationship to the rickettsial Wolbachia endobacteria of arthropods and to other filarial intracellular bacteria. The impact of tetracycline therapy of infected rodents on L. sigmodontis development was analyzed in order to understand the role(s) these bacteria might play in filarial biology. Tetracycline therapy, when initiated with L. sigmodontis infection, eliminated the bacteria and resulted in filarial growth retardation and infertility. If initiated after microfilarial development, treatment reduced filarial fertility. Treatment with antibiotics not affecting rickettsial bacteria did not inhibit filarial development. Acanthocheilonema viteae filariae were shown to lack intracellular bacteria and to be insensitive to tetracycline. These results suggest a mutualistic interaction between the intracellular bacteria and the filarial nematode. Investigation of such a mutualism in endobacteria-containing human filariae is warranted for a potential chemotherapeutic exploitation.

Mechanical resistance of the periorbita and the orbital floor complex--are isolated orbital floor fractures only a soft tissue problem?

The primary aims of orbital floor reconstruction are to prevent enophthalmos and herniation of the orbital contents in order to achieve correct globe position. Theoretically, the mechanical load of the orbital floor is approximately 0.0005N/mm(2) (30g orbital content onto 600mm(2) of orbital floor area). Therefore, low mechanical stress from orbital floor reconstruction materials is expected. The periorbita and orbital floor complex (bony orbital floor with periorbita) of 12 human cadavers were investigated for their mechanical resistance to distortion and compared to different absorbable pliable reconstruction materials after modification with pores (Bio-Gide, Creos, and PDS). The human periorbita resistance (approximately 1.4N/mm(2)) was comparable to that of the absorbable membranes (Creos, Bio-Gide), and the resistance of PDS (approximately 2.3N/mm(2)) was comparable to that of the orbital floor complex. The periorbita has a higher stability than the bony orbital floor. Therefore, in isolated orbital floor fractures with a traumatized bony orbital floor and periorbita, reconstruction of the soft tissue as a periorbita equivalent with a resorbable membrane appears to be adequate to prevent enophthalmos and herniation of the orbital contents.

Hysteretic behavior of the hepatic microsomal glucose-6-phosphatase system.

Carbamyl-P:glucose and PPi:glucose phosphotransferase, but not inorganic pyrophosphatase, activities of the hepatic microsomal glucose-6-phosphatase system demonstrate a time-dependent lag in product production with 1 mM phosphate substrate. Glucose-6-P phosphohydrolase shows a similar behavior with [glucose-6-P] less than or equal to 0.10 mM, but inorganic pyrophosphatase activity does not even at the 0.05 or 0.02 mM level. The hysteretic behavior is abolished when the structural integrity of the microsomes is destroyed by detergent treatment. Calculations indicate that an intramicrosomal glucose-6-P concentration of between 20 and 40 microM must be achieved, whether in response to exogenously added glucose-6-P or via intramicrosomal synthesis by carbamyl-P:glucose or PPi:glucose phosphotransferase activity, before the maximally active form of the enzyme system is achieved. It is suggested that translocase T1, the transport component of the glucose-6-phosphatase system specific for glucose-6-P, is the target for activation by these critical intramicrosomal concentrations of glucose-6-P.

Detection of the genial spinal canal in atrophic mandibles with a CBCT: a cadaver study.

OBJECTIVES: The use of a single midline implant to retain a complete mandibular denture when more implants cannot be used is an incipient treatment modality. However, in the mandibular symphysis, the genial spinal canal (GSC) is an anatomical structure with neurovascular content that can be harmed during dental implant surgery. The purpose of the present study was to use CBCT of edentulous atrophic cadaver mandibles and evaluate how often the simulated placement of a single midline implant would contact the GSC if present. METHODS: CBCT scans of 47 edentulous cadaver mandibles were performed. A digital simulation of the placement of a single midline implant (3.8 × 11.0 mm) was performed, and the implant-GSC contact was evaluated. RESULTS: A GSC was detected in the CBCT scan of all atrophic mandibles. In 42 cases (89.4%), the single midline implant contacted the GSC. On average, the five cases without GSC contact had a higher alveolar ridge (4.1 mm) and a lower GSC (0.79 mm) than did the cases with GSC contact. CONCLUSIONS: CBCT scans can adequately detect the GSC during pre-surgical diagnostics. There is a high risk of implant-GSC contact during surgery of the anterior mandible. However, the clinical relevance of such a contact is not known yet, because none of the clinical studies evaluating a single midline implant has reported any implant-GSC contact-related complications.

Intensive rearing of male calves during the first three weeks of life has long-term effects on number of islets of Langerhans and insulin stained area in the pancreas.

Permanent effects of early postnatal nutrition on the development and function of tissues and organs have been previously demonstrated primarily in humans and rodents. The objective of this study in calves was to analyze the impact of rearing conditions during the first 3 wk of life on morphology of insulin-producing pancreatic ß-cells. Forty-two male Holstein calves were raised during the first 3 wk of life either intensively (intensively reared [INT]; ad libitum milk feeding and individual hutches; = 21) or according to an established restrictive rearing protocol (4 L milk/d) during wk 1 in hutches and 720 g/d milk replacer (MR) from d 8 to 21 in group pens (restrictively reared [CON]; = 21). Thereafter, all calves were housed and fed under comparable conditions. Birth weight and weekly BW up to wk 10 were recorded. Plasma glucose, insulin, IGF-1, and GH levels were assessed in wk 1, 2, 3, and 10 of life. Slaughtering took place after 8 mo and pancreatic tissue from the medium body (corpus pancreatic) was removed. The number of islets of Langerhans and the insulin stained area were examined histologically. Total milk intake of INT calves was nearly double the intake in CON calves in the first 3 wk of life ( < 0.01). Daily starter intake during wk 4 to 10 of life did not differ between groups ( = 0.24). During the first 3 wk, the ADG were up to 9 times higher in INT calves compared to CON calves ( < 0.01), yet BW at time of slaughter did not differ ( = 0.18). Intensive rearing led to increased plasma glucose, insulin, and IGF-1 concentrations after 3 wk of life compared with rearing to the established standard protocol (all < 0.05), whereas GH was lower in INT calves during the second week of life. At time of slaughter, the mean number of islets of Langerhans was higher in INT calves compared to CON calves (9.1 ± 0.3 vs. 7.8 ± 0.3; < 0.01). Also, the total insulin stained area per photograph was higher in INT calves compared to CON calves (107,180 ± 4,987 vs. 84,249 ± 4,962 µm; < 0.01). Number of islets of Langerhans was negatively associated with birth weight but positively correlated with insulin and in trend with IGF-1 plasma levels during the second week of life. Insulin stained area tended to be linked with IGF-1 concentration during the third week of life. In conclusion, differences in the morphology of pancreatic islets of Langerhans indicate that calves can be programmed metabolically by an altered postnatal rearing intensity.

Riboflavin-mediated axonal degeneration of postnatal retinal ganglion cells in vitro is related to the formation of free radicals.

It is well known that glial cells produce several neurotrophic factors. We detected a neurogedegenerative/neurite growth inhibiting activity in serum-free astrocyte-conditioned medium (ACM). After high performance liquid chromatography (HPLC)-purification, spectral analysis and test of biologic activity in tissue cultures of postnatal retinal explants we isolated a fraction containing a riboflavin-(vitamin B2)-like compound which caused the neuronal degeneration. We therefore investigated the influence of pure riboflavin on axonal regeneration in vitro. Riboflavin is a normal compound of Dulbecco's modified Eagle medium (DMEM) and other tissue culture media in various concentrations. The removal of riboflavin from ACM by reversed phase chromatography abolished the neurite growth inhibiting effect and enhanced the regenerative response of axonal outgrowth from postnatal rat retinal explants. However, doubling of the normal medium concentration (1 microM) of riboflavin lead to strong degenerative alteration of the outgrowing axons in a dose-dependent manner, even under maximal growth stimulation by cultivating the explants in astrocyte-conditioned medium. To check the possibility that riboflavin-mediated cytotoxicity is related to the production of free radicals through photoabsorption from daylight, we irradiated culture medium with UV light, and induced radical stress by incubating the explants with Fe2+/3+. In an other set of experiments, we proofed, if antioxidants/free radical scavengers like pyruvate or vitamin C and E are able to overcome the neurite growth inhibiting influence of riboflavin or the radical stress. Our findings suggest an involvement of riboflavin-mediated formation of free radicals/reactive oxygen species and subsequent neurite degeneration in in vitro-assays of neuronal regeneration or neuronal cell cultures. How far the riboflavin/free radical-induced axonal degeneration could be an explanation for neurological degenerative disorders has to be elucidated.

A radioactive assay for the degradation of neuropeptide Y.

Neuropeptide Y (NPY) is one of the most abundant neuropeptides in the mammalian central nervous system. Like other neuropeptides, NPY is inactivated by specialized neuropeptidases. To trace the degradation of NPY, an assay was established using biotinylated NPY. Biotinyl-NPY was radiolabeled with Na125l by the chloramine-T method and bound to a streptavidin-agarose matrix. The amount of radiolabeling was analyzed by reverse-phase HPLC. The assay was carried out with five peptidases and inhibitors to demonstrate different specific activity. Measurable amounts of radioactivity were released by treatment with endopeptidase-24.18, plasmin, and trypsin, whereas dipeptidylpeptidase IV (DPPIV) and angiotensin-converting enzyme (ACE) showed no activity in this assay. In the case of DPPIV this is due to a resistance of the assay to aminopeptidase attack. The assay is useful to study the specific degradation of NPY particularly by endopeptidases in all kinds of biological samples.

Copper stained protein gels can be efficiently used for immunoblotting.

We have found that the previously described fast and sensitive copper staining of proteins resolved by sodium dodecyl sulphate-polyacrylamide gel electrophoresis does not interfere with the subsequent electrotransfer of these proteins to a solid support and their detection by specific antibodies. After the gel is copper stained and photographed it is simply destained and then equilibrated in transfer buffer prior to immunoblotting. We find that this treatment has no significant effect on transfer efficiency or band sharpness and is compatible with all common detection methods for the blotted proteins. It thus permits the separation of proteins to be checked in a simple way before immunoblotting is performed.