A video feedback-based tobacco cessation counselling course for undergraduates-preliminary results.

BACKGROUND: Hungary has one of the highest rates of tobacco use and decayed, missing and filled teeth in Europe, and the number of lung cancer-related deaths per annum is amongst the highest globally. As it is estimated that the rate of smokers who see their dentist or physician annually is about 70%, to involve all healthcare providers in tobacco intervention seems to be a promising strategy to reduce tobacco use in countries like Hungary. Such an intervention should obviously include the dental health team. It has already been suggested by experts on this topic that instruction in tobacco use prevention and cessation counselling for dental professionals and students of dentistry should be included in under- and postgraduate curricula. OBJECTIVES: To present a novel, video feedback-based undergraduate cessation counselling programme, which has recently been introduced to the dental curriculum at the Faculty of Dentistry, Szeged, Hungary. METHODS: Applying a problem-based learning approach, the programme consists of three main activities: a small-group interactive training session led by a faculty member, where students learn about the basic science and clinical aspects of tobacco use, including counselling skills; student interactions with professional actors (i.e. standardised patients) simulating real-life dental situations, which are recorded for post hoc evaluation; and finally an evaluation of the recorded performance of each student, with the participation of the actor, the student and a faculty member. RESULTS: With the help of this new approach, students had the chance to learn about and develop a deeper understanding of tobacco-related professional dental communication in realistic, case-based dental scenarios. Students have reported increased confidence in tobacco counselling after having participated in this programme. Furthermore, this method appears to be an ideal tool for the evaluation of both verbal and non-verbal tobacco counselling skills. CONCLUSION: To our knowledge, we are the first to have applied video feedback combined with behavioural modification methods in the teaching of tobacco cessation counselling. We conclude that teaching method can help dentists better understand smokers, gain confidence in tobacco cessation counselling and become more effective promoters of a smoke-free lifestyle. In addition, this method can be easily adapted to other healthcare educational settings, including other oral health training programmes.

Mental representations of the new organ and posttransplant patients' anxiety as related to kidney function.

Transplant patients' representations of their illness, body, and emotional state significantly influence their recovery. In this study, our primary aim was to examine the possible connections between emotional factors, body and illness representations, and renal function after 58 kidney transplantations. To measure mental representations of transplanted kidneys, we developed a projective drawing test. Other assessment instruments were the Beck Depression Inventory, Spielberger's State and Trait Anxiety Scale, and an in-house questionnaire. We also measured conventional kidney function markers, such as serum creatinine and urea levels. Analysis of our results revealed that patients with higher anxiety levels drew significantly larger kidneys in their projective drawing tests, and displayed significantly higher 10-day creatinine and urea level leading us to consider interrelations of an organ's intrapsychic integration and kidney function. If the graft is not integrated mentally in the body image, the representations of the "foreign body" can be associated with such psycho-neuro-immunologic processes of anxiety, which eventually may lead to adverse physiological effects on kidney function.

Paraneoplastic T cell lymphocytosis associated with a thymoma in a dog.

A four-year-old male neutered Australian shepherd dog was diagnosed with a thymoma and concurrent mature T cell lymphocytosis. The lymphocytosis consisted of a mixed population of T cells expressing either CD4 or CD8 or neither marker, and the result of polymerase chain reaction for antigen receptor rearrangement was negative. The peripheral lymphocytosis resolved within 24 hours following thoracotomy and thymectomy. Similar cases have been reported in man, but the aetiology of the increased circulating lymphocytes remains unclear. Although peripheral lymphocytosis is an uncommon paraneoplastic syndrome associated with thymomas, thymoma should be considered as a differential when the increased lymphocytes consist of a mixed population of T cells.

Genistein induces phosphorylation of cAMP response element-binding protein in neonatal hypothalamus in vivo.

We investigated the effect of the phytoestrogen, genistein and 17beta-oestradiol on cAMP response element-binding protein (CREB) phosphorylation in the neonatal female rat hypothalamus in vivo using western blot analysis and immunohistochemistry. Although CREB expression was insensitive to the compounds we tested, administration of genistein and 17beta-oestradiol induced rapid CREB phosphorylation (< 15 min) in the hypothalamus and its level remained elevated at 4 h. Quantitative immunohistochemical analysis showed that genistein and 17beta-oestradiol had no effect on CREB phosphorylation in the magnocellular subdivision of paraventricular nucleus. By contrast, genistein induced a dose-dependent increase in CREB phosphorylation in the medial preoptic area (mPOA) and anteroventral periventricular nucleus (AVPV). Administration of 17beta-oestradiol also caused a rapid, dose-dependent increase in CREB phosphorylation in the hypothalamus, mPOA and AVPV. These results demonstrate that genistein induces oestrogen-like rapid action on CREB phosphorylation in the neonatal central nervous system in vivo.

Cisplatin: a review of toxicities and therapeutic applications.

Cisplatin is a platinum chemotherapeutic used in a variety of malignancies. The antineoplastic activity occurs from DNA cross-links and adducts, in addition to the generation of superoxide radicals. Nephrotoxicity is the most well-known and potentially most clinically significant toxicity. Unfortunately, the mechanism for cisplatin nephrotoxicity has not been completely elucidated; however, many theories have been developed. Other toxicities include gastrointestinal, myelosuppression, ototoxicity and neurotoxicity. Saline diuresis is currently the most accepted way to prevent cisplatin nephrotoxicity. Research has focused on pharmaceuticals and enzyme/molecular alterations as alternatives to long-term diuresis. No agents have currently been identified that can protect from all toxicities. Cisplatin has shown activity against osteosarcoma, transitional cell carcinoma, squamous cell carcinoma (SCC), melanoma, mesothelioma, carcinomatosis and germinal cell tumours in the dog. In the cat, cisplatin cannot be utilized because of fulminant pulmonary oedema that occurs at standard doses. Intralesional cisplatin has been utilized in horses for the treatment of SCC and sarcoids.

Sex differences in oestrogen-induced p44/42 MAPK phosphorylation in the mouse brain in vivo.

In addition to the classical direct genomic mechanisms of action, oestrogen also exerts poorly understood, nonclassical effects on the signalling system in neurones. In the present study, we investigated whether sex differences exist in gonadectomy- and oestrogen-induced effects on p44/42 mitogen-activated protein kinase (MAPK) phosphorylation in specific brain regions of mice. We demonstrate that MAPK immunoreactivity was not altered by gonadectomy or oestrogen treatment in either sex. However, we show that the level of phosphorylated MAPK (pMAPK) within the anteroventral periventricular nucleus (AVPV) was consistently higher in males than females irrespective of gonadal steroid hormone status. In addition, gonadectomy was found to decrease pMAPK immunoreactivity within the piriform cortex of males. Oestrogen increased pMAPK immunoreactivity in the medial preoptic area and AVPV of females, but failed to have the same effect in male mice. Overall, these results demonstrate a marked sex difference in oestrogen-induced alteration of MAPK phosphorylation in the brain in vivo.

Facilitation of spike-wave discharge activity by lipopolysaccharides in Wistar Albino Glaxo/Rijswijk rats.

In normal rats the proinflammatory cytokines like interleukin-1beta, interleukin-6, which are induced by bacterial lipopolysaccharides, are able to control thalamo-cortical excitability by exerting strong effects on physiological synchronization such as sleep and on pathological synchronization like that in epileptic discharges. To investigate whether proinflammatory cytokines or lipopolysaccharides could modulate absence seizures resulting from a very different generator mechanism than the already investigated bicuculline-, kindling- and kainate-induced seizures, we used a genetically epileptic Wistar Albino Glaxo/Rijswijk rat strain, which is spontaneously generating high voltage spike-wave discharges. Wistar Albino Glaxo/Rijswijk rats responded with an increase of the number of spike-wave discharges to lipopolysaccharide injection (from 10 microg/kg to 350 microg/kg). Repetitive administration of 350 microg/kg lipopolysaccharides daily for 5 days increased the number of spike-wave discharges on the first, second and third days but the number of spike-wave discharges returned to the control value on day 5, at the 5th injection of lipopolysaccharides, showing a tolerance to lipopolysaccharides. The lipopolysaccharide-induced increase in spike-wave discharges was not directly correlated with the elevation of the core body temperature, as it is in febrile seizures, although lipopolysaccharide induced prostaglandin and is clearly pyrogenic at the doses used. Indomethacin, the prostaglandin synthesis inhibitor, efficiently blocked lipopolysaccharide-induced enhancement of spike-wave discharge genesis suggesting that the spike-wave discharge facilitating effect of lipopolysaccharides involves induction of cyclooxygenase 2 and subsequent synthesis and actions of prostaglandin E2. Low dose (40 mg/kg, i.p.) of competitive N-methyl-d-aspartate receptor antagonist 2-amino-5-phosphonopentanoic acid, and low dose of lipopolysaccharide (20 microg/kg) showed a synergistic interaction to increase the number of spike-wave discharges, whereas at supramaximal doses of lipopolysaccharide and the N-methyl-D-aspartate antagonist no synergy was present. The data reveal a functional connection between absence epileptic activity and lipopolysaccharide induction of prostaglandin synthesis and prostaglandin action and suggest some common cellular targets in epilepsy and lipopolysaccharide-induced inflammation.

Attitudes of Hungarian healthcare professional students to tobacco and alcohol.

Tobacco represents the single most preventable cause of disease and death in the world today. There were an estimated 3 million deaths annually at the end of the 20th century and it has been estimated that this will rise to more than 10 million by 2030. A disproportionate share of the burden of mortality is already being borne in the Russian Federation and the countries of Eastern Europe. For example, some of the highest rates worldwide of cigarette consumption and smoking prevalence are in the Czech Republic, Hungary and Poland. Cancer is one of the major tobacco-related causes of disease and death and Hungary has the highest male incidence rates in the world for both oropharyngeal and lung cancer. A consequence of this is that the mortality rate of middle-aged (50-60-year-old) men in Hungary today is higher today than it was in the 1930s, particularly among the lower socioeconomic groups. Of the many different approaches to tobacco cessation and control, advice or intervention by healthcare professionals ranks high in effectiveness; a recent survey indicated that requests from healthcare professionals to quit ranked second in effectiveness after requests by the smoker's own family. If healthcare professionals are to play a role in reducing death and disease from tobacco related cancers it is necessary to assess the attitudes and behaviours among healthcare professional students. Our survey assesses perceptions, behaviour and consequences among university students.

GRP-receptor-mediated signal transduction, gene expression and DNA synthesis in the human pancreatic adenocarcinoma cell line HPAF.

Bombesin-like peptides have been implicated as growth factors in various human cancers. Human adenocarcinoma cell lines (Capan-1, Capan-2, MiaPaCa-2 and HPAF) were tested to determine whether they express the gastrin-releasing peptide-preferring bombesin receptor (GRPR) and neuromedin B-preferring bombesin receptor (NMBR). Using RT-PCR the highest level of GRP receptor mRNA was found in HPAF cells. NMB receptor mRNA expression moderate in all cell lines investigated. We therefore selected the HPAF cell line to investigate whether bombesin treatment affects intracellular Ca(2+) ([Ca(2+)](i)), cAMP level, DNA synthesis as a measure of cell proliferation, and expression of three transcription factors: c-fos, c-myc and high mobility group protein IY (HMG-I(Y)).Bombesin administration led to an immediate increase in free intracellular Ca(2+) concentration ([Ca(2+)](i)) but did not change cAMP levels. The peptide also enhanced [(3)H]thymidine incorporation in HPAF cells (but not in the other cell lines), an effect that was concentration dependent, reaching 36 +/- 5% stimulation over control values at 24 h with an EC(50) of 2.27 x 10(-12) M. Furthermore, bombesin stimulated c-fos, c-myc and HMG-I(Y) expression in a time-dependent manner: the c-fos mRNA level increased dramatically in the first 30 min of exposure, then returned to basal level within 2 h, while the c-myc and HMG-I(Y) mRNA levels peaked at 2 h and 4h, respectively. All actions of bombesin were blocked by BME (D-Phe(6)-bombesin-(6-13)-methylester), a selective GRP receptor antagonist, but not by the NMB receptor antagonist BIM-23127 (D-Nal-cyclo[Cys-Tyr-D-Trp-Orn-Val-Cys]-Nal-NH(2)). We conclude that HPAF cells express mRNA for GRP receptors and that functional receptors are present in the cell membrane. The occupation of these receptors leads to a sequence of intracellular events involving rapid mobilization of intracellular Ca(2+), expression of c-fos, c-myc and HMG-I(Y) mRNA, and stimulation of cell proliferation. Conversely, although NMB receptor mRNA can be detected, its actual translation to functional receptors does not reach a detectable level.

Estrogen receptor-beta immunoreactivity in luteinizing hormone-releasing hormone neurons of the rat brain.

Feedback regulation of luteinizing hormone-releasing hormone (LHRH) neurons by estradiol plays important roles in the neuroendocrine control of reproduction. Recently, we found that the majority of LHRH neurons in the rat contain estrogen receptor-beta (ER-beta) mRNA, whereas, they seemed to lack ER-alpha mRNA expression. In addition, we observed nuclear uptake of (125)I-estrogen by a subset of these cells. These data suggest that ER-beta is the chief receptor isoform mediating direct estrogen effects upon LHRH neurons. To verify the translation of ER-beta protein within LHRH cells, the present studies applied dual-label immunocytochemistry (ICC) to free-floating sections obtained from the preoptic area of rats. The improved ICC method using the silver-gold intensification of nickel-diaminobenzidine chromogen, enabled the observation of nuclear ER-beta-immunoreactivity in the majority of LHRH cells. The incidence of ER-beta expression was similarly high in LHRH neurons of ovariectomized female (87.8 +/- 2.3%, mean +/- SEM), estradiol-primed female (74.9 +/- 3.2%) and intact male (85.0 +/- 4.7%) rats. The presence of ER-beta mRNA, ER-beta immunoreactivity and (125)I-estrogen binding sites in LHRH neurons of the rat provide strong support for the notion that these cells are directly regulated by estradiol, through ER-beta. The gene targets and molecular mechanisms of this regulation remain unknown.

[Automatic measurement of dye filling of simultaneous digital ICG- and fluorescein angiography sequences]. / Automatische Berechnung der Kontrastmittelfüllung simultaner digitaler ICG- und Fluoreszeinangiographiesequenzen.

BACKGROUND: The ICG filling is supposed to be faster than Fluorescein filling. Interestingly the filling characteristics of these dyes were never correlated directly using precise quantitative methods. Since ICG and Fluorescein are injected as a mixture, the simultaneous 2-channel angiography provides a suitable method to correlate the filling characteristics of the dyes. MATERIAL AND METHODS: The simultaneous ICG and Fluorescein angiograms were recorded with a Rodenstock Scanning Laser Ophthalmoscope. The angiographic images were digitized real-time with a graphic workstation. Filling characteristics of the two dyes was calculated after off-line eye tracking in different regions of interests (ROIs) on the central retina. RESULTS: The Fluorescein filling was faster than the ICG filling in 56.5% of our patients. In 26% of our patients was a mixed filling detectable. Depending on the position of the ROIs the Fluorescein or ICG filling was faster. In only 17.5% of our cases was the ICG filling faster than the Fluorescein filling. CONCLUSION: Our results show that Fluorescein filling in more than 50% of the cases is faster than ICG filling and only a minority of the patients has a faster ICG filling. According to our experience the filling pattern of the two dyes is individual, there is no rule of thumb for the filling.

Inhibitory effect of a long-acting somatostatin analogue on EGF-stimulated cell proliferation in Capan-2 cells.

Numerous studies have reported diverse effects of gut-derived regulatory peptides on growth of the normal pancreas, pancreatic neoplasms induced experimentally in animals, and pancreatic cancer cell lines, but the results of these investigations are rather controversial. The stimulatory effect of epidermal growth factor (EGF) on cell proliferation of pancreatic cell lines is well established. Whether this action can be modulated by somatostatin is not clear. Furthermore, it is not certain whether another regulatory peptide, cholecystokinin (CCK), affects the proliferation of these cells. In the present study we investigated the presence of CCK-A and CCK-B, as well as somatostatin-2 (SSTR2) receptors by RT-PCR, and studied the actions of EGF, CCK and octreotide on DNA synthesis in the human pancreatic adenocarcinoma cell line Capan-2. Octreotide, a long-acting somatostatin analogue was used as somatostatin agonist. Cells were cultured in RPMI-1640 medium. They were incubated in serum free medium containing 0.2% BSA in the absence (control) or the presence of the peptides. [3H]-thymidine incorporation into DNA was measured after 48 h of incubation. By means of RT-PCR analysis we were able to demonstrate SSTR2 expression, but not CCK-A or CCK-B receptor mRNA in Capan-2 cells. DNA synthesis evaluated by [3H]-thymidine incorporation was found to be increased by 45.2 +/- 5.6% in response to EGF (10(-8) M) and decreased by 11.7 +/- 2.6% to octreotide (10(-8) M) compared to controls (P < 0.01). The increase in [3H]-thymidine incorporation was significantly lower when EGF treatment was combined with octreotide administration (10.1 +/- 2.5% over control). In the concentration range of 10(-11)-10(-8) M, CCK did not alter significantly the incorporation of [3H]-thymidine into DNA in Capan-2 cells. In conclusion, these data support a role for EGF as a growth factor for the human pancreatic cancer cell Capan-2. Somatostatin may play an important role in regulating cell proliferation in Capan-2 cells both under basal, and growth factor-stimulated conditions. Our results suggest, however, that CCK receptors are not expressed, and CCK does not affect cell proliferation in this transformed pancreatic cell line.

Health risk behaviour of a medical student population: report on a pilot study.

A pilot study of a cross-sectional nature was carried out to observe and describe the health risk behaviour of a medical student population. The participants (242) were drawn from the students of the University Medical School of Szeged, Hungary. The students were aged 18-31 years (x = 23) and were randomly selected. The response rate was (73%). The project focussed on 4 harmful habits ranked in the following order of prevalence: excessive coffee drinking (35%), smoking (20.9%), regular alcohol use (6.8%) and illicit drug use (5.1%). The non-parametric (Chi-square) test showed significant differences between the higher and lower physical activity groups in terms of psychological well-being (p < 0.05) and health behaviour changes (p < 0.005). Harmful habits, however, were reported more frequently by the higher physical activity group. Significant differences could be detected in terms of women's illicit drug use (p < 0.05). Using the Mann-Whitney U-test, it was detected that those who performed more physical activities rated their health significantly higher (p < 0.001). This study will be pursued in an expanded study with a larger sample and concentrate especially on the relationship of physical activity behaviour to harmful habits. Follow-up methods are also planned to study the medical student population over time, which should yield some greater insight into these relationships.

[Epidemiology of psychosomatic symptoms and its effect on the self-evaluation of general health in university students]. / Pszichoszomatikus tünetek epidemiológiája és hatása az egészségi állapot önértékelésére egyetemi hallgatók körében.

This paper reports on the prevalence of some common psychosomatic symptoms as a part of a larger study of health state and health risk behaviour of a medical student population in Szeged. The prevalence of psychosomatic symptoms was considered as a health-related variable. In the study 691 students participated, the investigation was carried out by survey method, using self-completed questionnaire. In both sexes, backache and sleeping disorders were the most frequent symptoms. Furthermore, men reported stomach ache and palpitation in higher occurrence, while in women stress-related headache and chronic fatigue were the most common among the self-reported symptoms. The index of symptoms were significantly higher among women than men (p < 0.0001). Prevalence of psychosomatic symptoms proved an important variable affecting self-perceived health. The literature reviewed by the authors suggests that health state of medical students are significantly better than students of other colleges. Unfortunately, the morbidity and mortality data of physicians show inverse results among other intellectual populations. The authors suggestion is applying standardised method and cooperation among epidemiological teams working on this question.

Transferrin receptors associate with drug resistance in cancer cells.

Transferrin mediates its growth promoting properties through transferrin receptors, which are increased on cancer cells. We have studied drug-sensitive and drug-resistant cancer cells by using quantitative flow cytomerty with fluorochrome labelled transferrin and antibody to transferrin receptor, and we have found that drug-resistant cells have more transferrin receptors than drug-sensitive cells. This finding was confirmed and extended by using isotopically labelled ligand and by correcting for differences in cell surface areas. We also found that the number of transferrin receptors could be down-regulated by calcium channel blockers, and that such down-regulation diminished drug resistance, suggesting a hitherto unrecognised role for transferrin receptors in drug resistance.

Adriamycin conjugates of human transferrin bind transferrin receptors and kill K562 and HL60 cells.

Adriamycin (ADR) was coupled to human transferrin (TRF) by using a glutaraldehyde crosslinking method. The TRF-ADR conjugates were separated by column chromatography and the molar ratio of ADR to TRF (i.e., conjugation number) for the studied conjugates was found to be 1.2. Analysis in sodium dodecyl sulfate-polyacrylamide gels demonstrated that TRF-ADR conjugates with this molar ratio had the same mobility as native TRF and contained few aggregates. The ADR remained conjugated to TRF under conditions of decreased pH known to occur in many intracellular compartments, and analysis by spectrofluorometry revealed that the conjugated ADR retained its ability to intercalate DNA. The TRF-ADR conjugates were shown by flow cytometry to preferentially bind tumor cells and cell-bound conjugates were found to be laterally mobile within plasma membranes. The binding of TRF-ADR conjugates was determined to be saturable, and competition experiments done with both radioiodinated and fluorescein-labeled TRF-ADR conjugates demonstrated dose-dependent inhibition of conjugate binding by unlabeled TRF, indicating that TRF-ADR conjugates were bound by TRF receptors. Cytotoxicity studies performed with tritiated thymidine incorporation and tetrazolium reduction assays revealed that TRF-ADR conjugates inhibited the proliferation of both K562 and HL60 cells in culture more effectively than free ADR. Such conjugates could provide a delivery system for ADR that would target the drug and possibly diminish its dose-associated complications.

Flash-induced reduction of cytochrome b-559 by Q infB (sup-) in chloroplasts in the presence of protonophores.

Flash-induced, fast (t 1/2 ≈ 1 ms), reversible reduction of the high potential cytochrome b-559 (cyt b-559HP) was observed in chloroplasts in the presence of 2 µM protonophore, FCCP (carbonylcyanide p-trifluoromethoxyphenylhydrazone), CCCP (carbonylcyanide 3-chlorophenylhydrazone) or SF 6847 (2,6-di-(t-butyl)-4-(2',2'-dicyanovinyl)phenol). These protonophores promote autooxidation of cyt b-559HP in the dark (Arnon and Tang 1988, Proc Natl Acad Sci USA 85: 9524). No fast photoreduction could, however, be observed if the molecules were oxidized with ferricyanide in the absence of protonophores. This suggests that the molecules must be deprotonated to be capable for fast photoreduction.Photoreduction of cyt b-559HP was largely insensitive to DBMIB (2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone), but was inhibited by DCMU (3-(3,4-dichlorophenyl)-1,1-dimethylurea). With a train of flashes, no oscillation could be observed in the amplitudes of photoreduction. These data strongly suggest that cyt b-559HP is reduced by the semireduced secondary quinone acceptor (QB (-)) of Photosystem 2.

Cytotoxicity of a transferrin-adriamycin conjugate to anthracycline-resistant cells.

Conjugates of adriamycin coupled to transferrin by glutaraldehyde are cytotoxic to human promyelocytic (HL-60) and erythroleukemic (K562) cells. Growth inhibition of adriamycin-sensitive cells, as evaluated by thymidine incorporation and the MTT-assay, was higher for conjugates than for free adriamycin. The cytotoxicity toward adriamycin-resistant K562 and HL-60 cells was 3-fold and more than 10-fold higher, respectively, for the transferrin-adriamycin conjugate than for the free drug. The effect of the conjugate was dependent on its adriamycin content, i.e., on its conjugation number.

Transferrin conjugates of adriamycin are cytotoxic without intercalating nuclear DNA.

Studies of the biological chemistry of most anticancer drugs have revealed their cytotoxicity is expressed after the drugs have entered cells. It is thought that anthracycline antitumor drugs exert their cytotoxicity by entering cells, diffusing into nuclei, and inhibiting topoisomerase II and/or intercalating DNA base pairs. In order to deliver anthracyclines to transferrin (TRF) receptors on the plasma membranes of human tumor cells, we have prepared conjugates of adriamycin (ADR) with human TRF. These TRF-ADR conjugates were found to be stable at low pH and to exert more efficient cytotoxicity than free drug. By using spectrofluorometry, we found that the fluorescence of ADR within the conjugate was quenched by native DNA, demonstrating the presence of conformationally available drug to intercalate with nuclear DNA. However, fluorescence was not quenched when conjugate was reacted with viable cells, indicating that ADR did not reach the nucleus. Results of fluorescence microscopy experiments confirmed that free but not conjugated ADR reached the nuclei of viable cells, and TRF-ADR conjugates labeled with fluorescein isothiocyanate were found to initiate lateral diffusion as determined by patch and cap reactions. The involvement of TRF receptors was shown by flow cytometry experiments in which native TRF inhibited binding of fluorescein-labeled TRF-ADR conjugates. These data suggest that TRF-ADR conjugates mediate cytotoxicity by a mechanism other than intercalation with nuclear DNA. This mechanism, revealed by conjugating ADR to a TRF carrier, may not initiate complications such as cardiotoxicity and drug resistance.

Characterization of the anti-cancer activity of transferrin-adriamycin conjugates.

The anthracycline anti-cancer drug adriamycin (Adr) was coupled to human transferrin (Trf) by using a glutaraldehyde technique. The effect of Trf-Adr conjugates and unconjugated Adr on human cells was determined by using normal peripheral blood mononuclear cells and chronic myelogenous K562 cells. Cytotoxicity was determined by using an assay that measures the conversion of a tetrazolium salt (MTT) into a purple product (formazan) by mitochondrial dehydrogenases in viable cells. We found that free Adr at a concentration of 1 x 10(-7) had little effect on K562 cells, while Trf-Adr conjugates inhibited 75% of cellular activity. When normal peripheral blood mononuclear cells were tested against Trf-Adr conjugates, the 50% inhibitory concentration was found to be 1.4-1.7 x 10(-6) M, at which concentration greater than 85% of K562 cells were inhibited. Interactions of Trf-Adr conjugates with plasma membrane energy-producing systems are the proposed mechanisms of cytotoxicity.