Science ethics education part II: changes in attitude toward scientific fraud among medical researchers after a short course in science ethics.

PURPOSE: To determine the impact of the short science ethics courses on the knowledge of basic principles of responsible conduct of research (RCR), and on the attitude toward scientific fraud among young biomedical researchers. METHODS: A total of 361 attendees of the course on science ethics answered a specially designed anonymous multiple- choice questionnaire before and after a one-day course in science ethics. The educational course consisted of 10 lectures: 1) Good scientific practice - basic principles; 2) Publication ethics; 3) Scientific fraud - fabrication, falsification, plagiarism; 4) Conflict of interests; 5) Underpublishing; 6) Mentorship; 7) Authorship; 8) Coauthorship; 9) False authorship; 10) Good scientific practice - ethical codex of science. RESULTS: In comparison to their answers before the course, a significantly higher (p<0.001) number of students qualified their knowledge of science ethics as sufficient after the course was completed. That the wrongdoers deserve severe punishment for all types of scientific fraud, including false authorship, thought significantly (p<0.001) more attendees than before the course, while notably fewer attendees (p<0.001) would give or accept undeserved authorship CONCLUSION: Even a short course in science ethics had a great impact on the attendees, enlarging their knowledge of responsible conduct of research and changing their previous, somewhat opportunistic, behavior regarding the reluctance to react publicly and punish the wrongdoers.

Science ethics education part I. Perception and attitude toward scientific fraud among medical researchers.

PURPOSE: To assess the knowledge of basic principles of responsible conduct of research and attitude toward the violations of good scientific practice among graduate biomedical students. METHODS: A total of 361 subjects entered the study. The study group consisted mainly of graduate students of Medicine (85%), and other biomedical sciences (15%). Most participants were on PhD training or on postdoctoral training. A specially designed anonymous voluntary multiple-choice questionnaire was distributed to them. The questionnaire consisted of 43 questions divided in 7 parts, each aimed to assess the participants' previous knowledge and attitudes toward ethical principles of science and the main types of scientific fraud, falsification, fabrication of data, plagiarism, and false authorship. RESULTS: Although they considered themselves as insufficiently educated on science ethics, almost all participants recognized all types of scientific fraud, qualified these issues as highly unethical, and expressed strong negative attitude toward them. Despite that, only about half of the participants thought that superiors-violators of high ethical standards of science deserve severe punishment, and even fewer declared that they would whistle blow. These percentages were much greater in cases when the students had personally been plagiarized. CONCLUSION: Our participants recognized all types of scientific fraud as violation of ethical standards of science, expressed strong negative attitude against fraud, and believed that they would never commit fraud, thus indicating their own high moral sense. However, the unwillingness to whistle blow and to punish adequately the violators might be characterized as opportunistic behavior.

Radio- and chemotherapy variably affect the general immunocompetence of lung cancer patients.

OBJECTIVE: The evaluation of the effects of radiotherapy and chemotherapy on the immune status of lung cancer patients. EXPERIMENTAL DESIGN: Prospective nonrandomized study. SETTING: Hospitalized care. PATIENTS: 121 patients with unresectable non-small-cell lung cancer (Stage IIIb or IV), who were planned for radiotherapy (n = 81) or chemotherapy (n = 40). MEASURES: The relative and absolute numbers of blood T lymphocytes and monocytes, as well as the mitogen-induced proliferative response of the former, and phagocyting capacity of the latter cell subpopulation, were determined in patients before starting any therapy. In radiotherapy (RT)-treated group, the immune parameters were evaluated after 45 Gy and 60 Gy had been given. In chemotherapy (ChT)-treated group, the same parameters were determined three weeks after the 2nd and 4th cycle of ChT. RESULTS: The number and proliferative response of T lymphocytes were significantly (p < 0.001) lower, while the number and phagocyting capacity of monocytes were significantly (p < 0.001) higher in all patients before therapy, in comparison to the controls. After RT, the T cell number and proliferative response were significantly (p < 0.001) decreased, while the number of monocytes and their phagocyting capacity remained unchanged, when compared to the pretreatment values. Unlike RT, chemotherapy did not change any investigated parameter, except for the phagocyting activity of monocytes, which was significantly (p < 0.02) decreased, in comparison to the pretreatment value, after four cycles of ChT only. CONCLUSIONS: Two cancer treatment modalities--radio- and chemotherapy--variably affect the immune status of lung cancer patients. The initial great disturbances of general immunity parameters are further aggravated by radiotherapy. Unlike RT, chemotherapy exerts no suppression at all; on the contrary, it tends to normalize some of the parameters of cellular immunity of lung cancer patients.

The influence of surgery and anesthesia on lymphocyte functions in breast cancer patients: in vitro effects of indomethacin.

Surgical trauma and anesthesia may lead to the postoperative immunosuppression, the exact mechanism of which is still unresolved. Among various factors, the role of prostaglandine PGE2-mediated suppression was also proposed. We investigated the influence of surgery and two anesthetic regimes on lymphoproliferative response (LPR) to PHA and on NK cell activity (MTT) in breast cancer patients, as well as the effect of indomethacin, a PGE2 synthesis inhibitor, on these lymphocyte functions in vitro. In 36 previously untreated patients the lymphocyte functions were assayed before, 24 hours and seven days after the surgery. In regard to LPR, three distinct response patterns were observed: a) significant (p < 0.05) increase of initially lowered LPR; b) significant (p < 0.001) decrease of initially normal LPR 24 hours after operation, followed by normalization after seven days; c) no change of initially normal LPR. Indomethacin in vitro significantly (p < 0.05) enhanced the diminished LPR only before surgery, no effect being seen after the operation. The NK cell function was unaffected by surgery regardless the initial level of activity. Indomethacin had no effect on this lymphocyte function. There was no difference between the patient groups submitted to the different anesthetic regimes. In conclusion, our results show that surgical trauma variably affect the lymphocyte functions of cancer patients, the effect not being related to the particular anesthetic regime used. The PGE2-mediated suppression is not likely to be involved in postoperative immune function impairment.

Effect of T-activin therapy on indomethacin modulation of lymphoproliferative response in vitro of melanoma patients.

BACKGROUND: Several studies showed that PGE-mediated immunosuppression in cancer patients may be differentially affected by conventional oncologic therapy. METHODS: Since there is little evidence about the action of immunotherapy on this suppression mechanism, we investigated the effect of therapy with a thymic agent-T-activin, on in vitro modulation of lymphoproliferative response (LPR) by indomethacin. RESULTS: The results demonstrated that indomethacin added in vitro enhanced LPR in early stage melanoma patients before therapy. T-activin therapy as an adjunct to surgery improved this lymphocyte function; the post-therapy in vitro addition of indomethacin did not significantly affect mitogen response. However, in those patients whose LPR was insufficiently enhanced by immunotherapy (3/8), indomethacin had improved their lymphocyte response. In the control patient group treated by surgery alone, indomethacin significantly enhanced LPR in vitro six months after operation. Although obtained in a small number of patients, our results indicate that the enhancing effect of T-activin therapy on lymphoproliferative response may be, at least in part, due to the effect on PGE-mediated suppressor cell activity. CONCLUSIONS: Furthermore, post-therapy in vitro testing may indicate a possible usefulness of this drug combination in some of the early stage melanoma patients.

Immunomodulation in vitro. The predictive value of in vitro testing of lung cancer patients' lymphocyte responsiveness to stimulation by Thymex L. A preliminary report.

Eleven lung cancer patients were selected for combined radio and immunotherapy with a thymic agent-Thymex L. The selection criteria included the pre-therapy testing of patients' immunocompetence and the responsiveness of their lymphocytes to the in vitro addition of Thymex L: only patients with a significant degree of immunodepression, whose depressed cellular immunity parameters (the number of total and active T cells and their mitogen-induced lymphoproliferative response) were significantly increased upon this agent's action in vitro, entered the study. The results of the pre-therapy in vitro stimulation correlated with those obtained after completion of radioimmunotherapy: the administration of Thymex L along with radiotherapy seances prevented iatrogenic deterioration of initial depression of general immunocompetence and enabled to overcome it to a certain degree. This indicates that pre-therapy in vitro testing has a true predictive value. However, the initial immune disturbances were not normalized by immunotherapy; the post-therapy testing of the patients' lymphocytes to the addition of Thymex L in vitro showed that these cells possessed a residual potential to respond by a significant increase of the active T cell number and proliferative capacity, suggesting that immunotherapy could be prolonged in order to potentiate cellular immunity in immunodepressed lung cancer patients.

The involvement of immune reactions in cardiac damage during acute myocardial infarction: role of cell-mediated immune response.

This study was undertaken with the aim of investigating humoral and cell-mediated immune response in acute myocardial infarction (AMI) as possible mechanisms involved in the infarction enlargement. Twenty three patients with first AMI and 15 healthy volunteers were examined. Of the AMI patients, 14 had extensive infarction (group A), while 9 patients had small infarction (group B). Immunologic analyses were performed at admission, and repeated after 3, 7, 14 and 21 days of the acute event. Following parameters were tested: number of CD3+, CD4+, CD8+ and CD20+ cells; serum IgG, IgA, IgM, C3, C4, immune complex and anticardiac antibody levels; polymorphonuclear cell (PMN) function (chemotaxis, phagocytosis, metabolic activity); leukocyte migration in vitro in the presence of water-soluble homologous heart extract. It was demonstrated that the number of B cells, serum IgG, C3, immune complex and anticardiac antibody levels were elevated from 7th-14th days after AMI. Concerning these parameters, however, no significant differences were obtained between group A and group B of AMI patients. Chemotaxis and metabolic activity of peripheral blood PMN, but not phagocytosis, were enhanced during AMI, again changes of PMN did not correlate with the extension of infarction. In contrast, leukocyte migration inhibition in vitro revealed that only patients with extensive AMI have developed positive reaction during the first 14 days after the onset of the disease, while leukocyte inhibition reaction appeared in patients with nonextensive AMI not earlier than the 21st day after the infarction. These findings demonstrate generation of immune reactivity during AMI and indicate that humoral immune response seems more likely to be an epiphenomenon related to tissue necrosis, while cell-mediated immune reactions could influence the extensiveness of cardiac damage.

Vitamin C inhibits random migration of malignant pleural effusion mononuclear cells.

Although many epidemiological studies indicate protective effect of vitamin C against a variety of human malignancies its mechanism(s) of action is questionable. The presented results show that the part of its effect may be accomplished by mononuclear cells, as necessary participants in body defence. Namely, in a long-term in vitro assay we tested vitamin C influence on random migration ability of malignant pleural effusion mononuclears (PEM) obtained from breast cancer patients. Vitamin C in a dose- (50-500 micrograms) and time-dependent (4-44 h) manner inhibited PEM motility, suggesting that immobilization of cells in situ may contribute to its beneficial effect in human cancers.

Clinical and immunologic effects of T-activin therapy in early stage melanoma patients.

We investigated the clinical and immunological effects of T-activin therapy in early stage melanoma patients. Several immune parameters (the number of T cells-E-RFC and CD3+, their subsets-CD4+ and CD8+, the number of CD38+ and CD16+ cells, and mitogen-induced lymphoproliferative response-LPR) were analyzed in relation to the clinical course of the disease in patients treated by T-activin in addition to the surgery (n = 8), and in control patients treated by the surgery alone (n = 9). Immunological tests were performed before therapy and one month after the last (6th) cycle of T-activin, i.e. six months after surgery in controls. The patients were followed-up from February 1991 to August 1995. Clinical evaluation showed that disease-free interval for observed period was similar in both groups of patients (17.5 and 13 months), while the survival time was longer in T-activin-treated patients than in controls (40 vs. 24 months), although this difference was not significant. The phenotyping analysis of peripheral blood lymphocytes showed no changes of the pretreatment values of total T cells and their subpopulations regardless the clinical course of the disease in both groups of patients. The number of NK cells (CD16+) was significantly increased after T-activin therapy, but this increase was not associated with clinical benefit, since it was seen in patients with the progression of the disease. In control patients, the initial number of CD16+ cells did not change significantly, irrespective of the clinical course. The lymphoproliferative response increased significantly in 4 out of 5 T-activin-treated patients with the progression of the disease, while a slight increase of this lymphocyte function was seen in 3 disease-free patients. In patients treated by surgery alone, especially those with disease progression, the LPR was significantly decreased six months after tumor excision. These findings, although obtained in small number of patients, suggest an immunomodulatory action of T-activin therapy in early stage melanoma patients, which did not correlate with the clinical course of the disease. On the other hand, an almost doubled survival time in T-activin-treated patients in comparison to the controls, may indicate a possible effect of T-activin therapy on some other immune functions not evaluated in this study. Further investigations in a larger number of patients is needed for assessment of the true effectiveness of such therapy.

[In vitro reactivity of lymphocytes after the addition of immunostimulatory drugs]. / Reaktivnosti limfocita in vitro na dodavanje imunostimulacionih sredstava.

The in vitro immunomodulating, effects of two thymic extracts--Thymex L and Thymomodulin--on lymphocytes of lung cancer patients, were studied. The number of total and active T cell and PHA-induced lymphoproliferative response were evaluated before and after the addition of two different concentrations (5 mg/ml and 0.5 mg/ml) of these agents. The in vitro incubation with Thymex L and Thymomodulin had no effect on the number of total T lymphocytes, but the active T cells and proliferative responses were significantly higher. This increases was observed in the majority of patients with diminished baseline values. These results indicated that several immunological parameters should be used in vitro testing of an immunomodulatory drug. They also indicated the need for in vitro testing selection of candidates for immunotherapy among immunodepressed cancer patients.

Continuous versus split-course irradiation for lung cancer. Immunological implications.

The therapeutical irradiation for lung cancer causes profound disturbances of host's general immunocompetence, the cellular immunodepression being the dominant finding. It is thought that split-course technique holds certain advantage over the continuous irradiation, since the former includes an interruption of 4 week duration, thus allowing the lymphopoietic system to recover to a certain degree. In this report, we compared the radiotherapy-due alterations of several parameters of cellular immunity (the number and function of total T cells, active T cells and the cells of monocyte/macrophage lineage), immediately after the completion of therapy in either continuously (n = 13) or split-course-irradiated (n = 12) lung cancer patients. All patients had received the total dose of 60 Gy. Both therapeutical techniques caused alterations of the parameters tested: the significant decrease of the total and active T cells and their proliferative responses, while the phagocytic activity and the number of mononuclear phagocytes were increased, the latter being affected to a lesser extent in split-course-treated patients. Our results suggest that both techniques have similar immunodepressant effect on the cellular immunity of lung cancer patients.

Immunosuppressive and cytogenetic effects of pelvic irradiation on the peripheral lymphocytes of patients with cervical cancer. One year follow-up.

The circulating lymphocytes of patients treated for cervical cancer were examined by four independent manners: by evaluation of T-cell proportion in peripheral blood, proliferative response upon PHA stimulation, PHA-induced leukocyte migration inhibition, and by concomitant chromosome aberration frequency. The immediate and longer-term effects of pelvic irradiation on T lymphocytes were investigated in 19 patients prior to, during, and immediately after radiotherapy, and then at subsequent intervals of two, three and five months. Radiotherapy caused profound depression of already diminished T-cell number and their proliferative response; both parameters gradually recovered during post-treatment period, and achieved their pretreatment values at the end of follow-up. The leukocyte migration inhibition was much less affected; it slightly deteriorated in the middle of post-treatment period, but reached the pretreatment level at the end of monitoring. The chromosome aberration frequency increased during irradiation in dose-dependent manner; it decreased gradually thereafter, but remained high during follow-up. Their elimination rate correlated with the recovery of T-cell number and proliferative response. However, at the end of monitoring, when all immunological parameters were completely recovered from harmful effect of irradiation, the percentage of chromosome aberrations remained high (12.5%), although significantly lower than the post-treatment one.

Immune reactive proteins in patients irradiated for cervical cancer. One year follow up.

The parameters of nonspecific humoral immunity--serum immunoglobulins and immune complexes--were evaluated in irradiated group of patients with uterine cervix carcinoma (Stages IIB and IIIB), during one year follow up. The concentrations of IgA, which were elevated in patients before therapy, slowly declined after the radiotherapy, at the end of the follow up being returned to the normal range. The concentrations of IgG, which were significantly decreased in patients immediately after the therapy, were sharply and transiently elevated two months after the therapy; afterwards, the levels of IgG gradually decreased to the values which did not differ from their pre- or post-RT levels, as well as from control ones. The levels of CIC were not significantly changed after radiotherapy, although a transient increase was found seven months after radiotherapy, being not different from controls at the end of the follow up. The serum IgM remained in the range of control values immediately after radiotherapy, as well as during monitoring. The slow normalization of the serum IgA and CIC levels might reflect the success of the therapy.

Enhancement of phytohemagglutinin-induced lymphoproliferative response by indomethacin, Thymex L or their combination in lung cancer patients.

Several studies showed that thymic factors and prostaglandin synthesis inhibitors enhance in vitro lymphoproliferative response (LPR) to mitogens in cancer patients. In this study we investigated whether indomethacin and thymic extract (Thymex L), applied in combination, may in a synergistic pattern influence phytohemagglutinin-induced LPR in lung cancer patients. The results demonstrate that the use of the investigated agents enhances LPR to a similar level in hyporeactive patients before, as well as after, therapy. However, this drug combination exerts an additive effect on LPR, but only in patients who underwent cytoreductive radiation therapy, indicating the potential usefulness of this drug combination as an adjuvant treatment of these patients.

Humoral immunity in patients with carcinoma of the uterine cervix--effects of radiation therapy.

The parameters of nonspecific immunity-serum immunoglobulins and immune complexes--were evaluated in patients with carcinoma of the uterine cervix (Stages IIB and IIIB) prior to, during and immediately after pelvic irradiation. In untreated patients, significantly elevated circulating IgA was found only in patients in Stage IIIB; serum IgG and IgM in both groups did not differ from control values. The level of circulating immune complexes in both groups was higher than in controls. Radiotherapy did not affect significantly any of the parameters examined; only the percentage of patients with elevated concentrations of IgA and IgG decreased during the treatment. These results showed that fractionated pelvic irradiation did not affect B cell function, these cells being more radioresistant than other lymphocyte subpopulations.

In vitro effect of indomethacin on mitogen-induced lymphoproliferative response in lung cancer patients.

There is some evidence that prostaglandin (PGE)-secreting cells may have a role in immunosuppression in cancer patients. In this work we investigated the effects of indomethacin--a PGE synthesis inhibitor, on PHA-induced lymphoproliferative response in vitro. Twenty patients with lung cancer before therapy were included in this study. When compared to controls, the patients had significant decrease of T cell number and proliferative response to PHA (p less than 0.001) and increased number of mononuclear phagocyting cells (p less than 0.001). The degree of depression of lymphocyte response did not correlate with the number of mononuclear phagocytes. The presence of indomethacin in the culture induced significant (p less than 0.01) improvement of the reactivity in high percentage (75%) of patients with diminished lymphoproliferative response to PHA. In the patients with normal lymphocyte response, indomethacin did not change reactivity to PHA. These results indicate that PGE-secreting cells may contribute to the immune depression in lung cancer patients, and that indomethacin may have therapeutical potential in some patients.

The protective activity of Thymex L against radiotherapeutically-induced cellular immunodepression in lung cancer patients.

In order to prevent the radiotherapeutically-induced aggravation of initial immunodeficiency, a thymic preparation (Thymex L) was given to lung cancer patients simultaneously with irradiation. The parameters of both cellular and humoral nonspecific immunity were evaluated in two groups of patients: one was treated with radiotherapy only (60 Gy in 30 fractions); the other one received Thymex L (100 mg 3 times a week, total dose 1800 mg, i.m.) simultaneously with radiotherapy. The significant decrease of B and T cell number, and decreased lymphoproliferative response to PHA were found in all patients before therapy; the number and phagocyting capacity of blood monocytes, as well as the concentrations of circulating IgG, IgA and immunocomplexes, were all significantly increased. Immediately after irradiation the patients had even lower number of T and B cells, diminished reactivity to PHA and higher number of mononuclear phagocytes when compared to the values before therapy. In patients treated with Thymex L, the number of B and T cells and PHA-induced proliferative response were significantly higher than in those treated with radiotherapy only. No effect of this therapy was seen on active T cells, on high number and function of mononuclear phagocytes and on elevated concentrations of serum immunoglobulins and immune complexes. Our results indicate that Thymex L can successfully prevent the harmful effect of radiation therapy on cellular immunity in a majority of lung cancer patients.