[Are herbs safe during pregnancy and breastfeeding].

The review rejects the popular understanding of the security and potential benefits of herbal products for pregnant or lactating women. The survey does not claim to be exhaustive, but presents a number of publications concerning cases of proven harmful impact after use of herbal preparations during pregnancy and breastfeeding. Most of these studies come from developing countries and relate to Asian herbal products that are increasingly marketed and used in Europe. Many of the cited authors believe on the importance of the proper information making prudent the practitioners and patients about the risks of use of plant products, especially during pregnancy and lactation. While definitive data emerge, all herbal products should be viewed with caution, although there are no explicit warnings that herbals should not be used during pregnancy and lactation. This especially refers to the usage of herbs during the first trimester of pregnancy. It is essential that patients are informed about the safety issues of herbals. After all, pregnancy is not a pathological condition and therefore it does not require treatment. So the debate about the benefit of food supplements and herbal remedies is beyond the rational need to use them for specific indications during pregnancy.

Telematics applications to support the role of the community pharmacists as self-medication advisors. TESEMED Consortium.

One of the applications developed within the EU-funded projects TESEMED and TESEMED-II is a program for the information and continuous training of community pharmacists, with the aim to empower them as advisors of the citizens about self-medication topics. Several programs are being developed on the basis of ad-hoc developed protocols about minor ailments (currently, cold and flu, haemorrhoids, constipation and cough). Each program includes three modules: a hypertextual version of the protocol, an interactive scheme of it, and an educational tool called Encounter Simulator, that trains the pharmacist about the protocol by means of simulated pharmacist-customer interactions. The testing of these applications with 84 community pharmacists offers positive results in terms of expectations, program characteristics and perceived usefulness.

Autoantibodies against a kidney--liver protein associated with quinolone-induced acute interstitial nephritis or hepatitis.

In the present study we report on four cases of acute interstitial nephritis (AIN) and two cases of hepatitis induced by quinolone. We show by immunoblotting analysis that all sera from these patients contained autoantibodies that recognize a 65-kDa protein expressed in normal human kidney and liver microsomes. Only 6% of sera from healthy individuals who did not ingest quinolone recognized the same protein. These findings suggest that the presence of autoantibodies could be used as a sensitive marker and that a modification of microsomal proteins by quinolone itself or by a metabolite could generated an autoimmune response.

Monitoring of azathioprine-induced immunosuppression with thiopurine methyltransferase activity in kidney transplant recipients.

Thiopurine methyltransferase (TPMT), one of the enzymes involved in azathioprine metabolism, exhibits a wide range of activity in the normal population. We prospectively evaluated the monitoring of erythrocyte TPMT activity (using a radiochemical method) in kidney transplant recipients with regard to the efficacy of azathioprine. Three patterns in TPMT activity variation were observed. In group 1 patients, TPMT activity rose as early as 8 days after transplantation and steadily until month 3. In group 3 patients, TPMT activity remained unchanged. In group 2 patients, TPMT activity rose at month 1 after transplantation. Interestingly, the incidence of acute rejection was significantly (P < 0.01) different among the 3 groups, with the lowest incidence in group 1 and the highest in group 3. We hypothesized that TPMT activity increase was induced by azathioprine in the patients with the lowest incidence of acute rejection. The inducibility of TPMT activity would then appear to be an interesting marker of azathioprine-induced immunosuppression.

Anti-liver microsomes autoantibodies and dihydralazine-induced hepatitis: specificity of autoantibodies and inductive capacity of the drug.

Anti-liver microsomes (anti-LM) autoantibodies in patients with dihydralazine-induced hepatitis were found to react specifically with cytochrome P4501A2 (P4501A2) but not with P4501A1 expressed in yeast and bacteria. These results were confirmed by immunoinhibition of methoxyresorufin-O-demethylase activity (supported by the P4501A subfamily); anti-LM antibodies more strongly inhibited this activity in yeast expressing P4501A2 than in yeast expressing P4501A1. Anti-LM were shown to be specific to the disease; in three cases, these autoantibodies were present at high titers during disease, whereas the titers decreased upon recovery and became undetectable a few months after recovery. Thus, there exists a time-dependent relationship between the disease and the autoantibodies, which does not prove that the autoantibodies are causative of the hepatitis; they might only be a marker. The inductive capacity of dihydralazine toward P450 was also studied. In rats treated in vivo and in human hepatocytes treated in vitro with dihydralazine, a 2-fold increase in P4501A2- and P4501A-supported monooxygenase activities was found. The levels of the other P450 isoforms tested were unchanged during treatment, both in vivo in rats and in vitro in cultures of human hepatocytes. In human hepatocytes, dihydralazine produced a dose-dependent increase in the level of P4501A up to 0.1 mM; induction of P4501A was less strong at 0.2 mM and disappeared at 0.5 mM. The same treatment did not change the level of P4503A4, taken as control. The strong heterogeneity in the expression of P4501A enzymes in human liver and the capacity of these enzymes for induction by dihydralazine and by other compounds might be predisposing factors in this autoimmune disease.

Enhancement of adriamycin cytotoxicity in sensitive and resistant sublines of human tumor cells by calcium antagonists.

The effect of the calcium antagonists cepharanthine and verapamil on adriamycin-induced cytotoxicity against sensitive (K 562 and Ov 2780) and resistant (K 562/ADM and AD 10) sublines of human tumor cells was evaluated. Nontoxic concentrations of cepharanthine moderately enhanced adriamycin cytotoxicity against sensitive sublines (2.1-2.5 fold). A significant enhancement (13-26 fold) of drug cytotoxicity was observed when resistant cells were treated with a combination of cepharanthine and adriamycin. The calcium influx blocker verapamil (used for comparison) also enhanced adriamycin cytotoxicity, although to a lesser extent. The fact that enhancement was 6-10 fold greater in resistant then in sensitive cells, as well as the loss of biphasic properties of adriamycin on dose-response curves after combined treatment, indicate that cepharanthine may play a role in overcoming drug resistance in some tumor cells.

Ryodipine-induced enhancement of farmarubicin cytotoxicity against human leukemia cells in vitro.

Ryodipine is a recently developed dihydropyridine calcium channel blocker, chemically similar to nifedipine but with some advantages: light stability, low toxicity, etc. In recent years calcium antagonists have attracted attention not only with their cardiovascular activity but also as drugs increasing the antitumor effect of some cytostatics. In the present work the potentiating effect of ryodipine on the farmarubicin-induced cytotoxicity against K 562 human leukemia cells in vitro was evaluated. The cells were exposed to farmarubicin, ryodipine and combination of both drugs. The results obtained clearly demonstrated that a nontoxic concentration (0.5 microgram/ml) of ryodipine increased (about 5 times) farmarubicin cytotoxicity as the IC50 of the cytostatic was shifted from 1,86 ng/ml on single treatment to 0.37 ng/ml on combined treatment of the cells. The data presented suggest a role for ryodipine in the enhancement of the antitumor activity of some cytostaticts.

Effect of calcium influx blocker verapamil on adriamycin-induced cytotoxicity in leukemia cells in vitro.

The effect of verapamil (calcium influx blocker) on adriamycin-induced cytotoxicity against sensitive and resistant subline of K 562 acute myelogenous human leukemia cells has been evaluated. Verapamil by itself at a concentration of 0.5 microgram/ml did not affect the cell growth. It was found that the nontoxic concentration of verapamil moderately enhanced adriamycin cytotoxicity against K 562 cells, showing enhancement ratio of 1.3-1.4 according to the schedule used. A significant enhancement of adriamycin cytotoxicity (enhancement ratio of 5.8) was observed when verapamil and adriamycin were administered simultaneously against resistant subline of K 562/ADM cells. The results obtained give grounds to assume that verapamil could be used to overcome drug resistance in tumor cells.

On the in vitro cytotoxicity of 1-chloroethyl-2-chloromethyl-2-imidazoline hydrochloride.

The cytotoxic activity of the monofunctional alkylating agent 1-chloroethyl-2-chloromethyl-2-imidazoline hydrochloride (chlorimidazine), synthesized according to predetermined properties, is characterized. The data obtained show that the substance has a cell-cycle state nonspecific cytotoxic effect. The interaction of chlorimidazine with cellular DNA is traced. It is found that the substance does not induce a break in the macromolecule, apurine sites or intercalation. Ensuing from the chemical structure of the compound, the probable mechanism of its cytotoxic action is discussed.

Dependence of bleomycin-induced cytotoxicity and DNA damage on the in vitro culture growth phase of mouse tumour cells.

EMT 6/Ca/VJAC cells have been treated with various doses of bleomycin and the effects of these treatments in terms of cell killing and DNA damage have been determined. The experiments have been carried out with exponentially growing and plateau-phase cultures. Dose-response curves have been obtained using a clonogenic cell survival assay. Bleomycin-induced DNA damage was quantitated by a fluorescence procedure, based upon the time-dependent partial unwinding of cellular DNA under alkaline conditions. Although cells in exponential-phase of culture growth were more sensitive to bleomycin than those in plateau-phase, this could only be attributed to enhanced drug-DNA interaction at high (greater than 40 micrograms/ml) drug concentration. Thus factors other than DNA damaging potential and related to culture growth status control drug sensitivity.

Potentiation of sensitivity to bleomycin and the drug-induced DNA damage in EMT6 cells by the calmodulin inhibitor trifluoperazine.

It has been shown that inhibitors of calmodulin can increase the sensitivity of rodent cells to Bleomycin by interfering with DNA repair functions. As a result of this study we have found that treatment of plateau-phase EMT6 cells with Bleomycin causes a decrease in cell survival due to DNA damage. This toxic effect can be potentiated by the addition of a nonlethal dose of the calmodulin inhibitor Trifluoperazine. Furthermore, it was found that Trifluoperazine greatly potentiated Bleomycin-induced DNA breakage. Considering the results obtained by alkaline-denaturation assay (detects single-strand breaks, including these arising from alkali-labile lesions) and nucleoid sedimentation (in neutral sucrose gradients) assay (detects only frank breaks), a difference, presumably due to the Trifluoperazine-produced discrete block in the repair of Bleomycin-induced DNA alkali-labile lesions was found. These data suggest a role for calmodulin in the DNA repair pathway and provide a rational basis for the use of calmodulin inhibitors in cancer chemotherapy.

Interaction of bleomycin, hyperthermia and a calmodulin inhibitor (trifluoperazine) in mouse tumor cells.

To improve the efficacy of thermochemotherapy we have investigated the individual and combined effects of hyperthermia (44 degrees C) and the calmodulin inhibitor trifluoperazine (30 micrograms/ml) on early plateau phase cultures of mouse EMT6 cells for simultaneous exposures to bleomycin. We found that a non-toxic combination of hyperthermia and trifluoperazine: enhanced the cytotoxicity of bleomycin, increased the frequency of long-lived attachment sites of cellular DNA at the nuclear matrix, and resulted in an accumulation of DNA damage (strand-breaks and alkali-labile lesions) caused by the inhibition of strand-break rejoining and the impaired processing of DNA sites involving base loss. Our findings implicate a role for calmodulin in the control of chromatin structural changes during DNA repair and the study provides a rational basis for the use of calmodulin inhibitors in thermochemotherapy.

The effect of a normal tissue radioprotector Adeturone on the radiation sensitivity of Lewis lung carcinoma: an enhanced response.

The effect of the normal tissue radioprotector Adeturone (S-2-aminoethylisothiuronium adenosine-5'-triphosphate) on the response of primary and metastatic Lewis lung carcinoma to radiation was evaluated. It was found that this compound enhanced the antitumor effect of whole-body and local radiation. In experiments with whole-body radiation Adeturone eliminated the lethality due to radiation toxicity (for a daily treatment schedule--10 X 1 Gy/fraction) or reduced it from 50% to 25% (for an intermittent schedule--3 X 3.34 Gy/fraction). Following local radiation treatment combined with Adeturone, a higher TWI%, longer tumor doubling time and longer mean survival time were recorded than for either treatment alone. An enhanced activity against metastatic growth of Lewis lung carcinoma to the lungs was also found with combined therapy.

Modification of the balance between damage and repair of bleomycin-induced DNA-strand breakage by hyperthermia.

It is well recognized that hyperthermia can greatly enhance the cytotoxic effect of bleomycin. It has been suggested that the mechanism of hyperthermic potentiation of bleomycin toxicity involves the inhibition of DNA repair function. We have investigated how hyperthermia can modify the repair capacity of bleomycin treated cells by monitoring the induction of DNA damage and repair, using two essentially different techniques (DNA unwinding assay and nucleoid sedimentation assay). We have also investigated whether the enhancement of DNA-damaging potential of bleomycin was related to the availability of new sites within chromatin for drug attack. We have found that heat treatment (even for non-toxic exposures of 30 min at 44 degrees C) restricts the access of active drug to DNA, probably by the increase in nuclear protein content. The results obtained show that the increase in drug-induced DNA damage in heat-treated cells probably represents an underestimation of the predominant effect of hyperthermia on reducing cellular DNA repair capacity.

Interaction of bleomycin, hyperthermia and a calmodulin inhibitor (trifluoperazine) in mouse tumour cells: II. DNA damage, repair and chromatin changes.

We have reported in the preceding paper that the treatment of plateau phase mouse EMT6 tumour cells with a combination of hyperthermia (HT; 44 degrees C) and trifluoperazine (TFP; 30 micrograms ml-1; an inhibitor of calmodulin) greatly enhances the cytotoxicity of the antitumour drug belomycin (BLM). The cytotoxic action of BLM is thought to arise from the induction of DNA damage in a manner which reflects chromatin accessibility. Thus we have studied the effects of the two modifiers (HT and TFP) on chromatin structure and BLM-induced DNA damage. Co-treatment of cells with HT and TFP altered chromatin organisation by the formation and slow resolution of new DNA attachment sites at the nuclear matrix. HT increased drug-induced DNA damage (strand breaks and alkali-labile lesions) by the general depression of repair rather than through the generation of new sites for drug action. TFP produced a more discrete block in the repair of alkali-labile lesions in DNA. Both processes appear to occur for the combination of BLM, HT and TFP, and we propose that the novel chromatin configuration permits the accumulation of potentially lethal DNA strand breaks. Our study indicates the potential value of chromatin/DNA repair modifying regimens for overcoming the poor responsiveness of some tumour cells to chemotherapeutic drugs and provides a rational basis for the use of calmodulin inhibitors in thermochemotherapy.

Interaction of bleomycin, hyperthermia and a calmodulin inhibitor (trifluoperazine) in mouse tumour cells: I. In vitro cytotoxicity.

Evidence in the literature suggests that hyperthermia (HT) or inhibitors of calmodulin can increase the sensitivity of rodent cells to bleomycin (BLM) by interfering with DNA repair functions. In an attempt to explore methods of improving the efficacy of thermochemotherapy we have investigated the individual and combined effects of HT (44 degrees C) and the calmodulin inhibitor trifluoperazine (TFP, 30 micrograms ml-1) on early plateau phase monolayer cultures of mouse EMT6 tumour cells for simultaneous exposures to BLM. Early plateau phase cultures are relatively resistant both to HT and to BLM. The selected HT and TFP regimens (either alone or in combination) were non-toxic. Comparing the sensitizing effect (given by the ratio: Do BLM/Do BLM + modifier) of the various regimens on BLM-treated cells, we found that: TFP alone had a marginal effect (ratio 1.3), HT alone showed significant potentiation (ratio 19) and the combination of HT and TFP strongly sensitized (ratio greater than 110) cells to BLM cytotoxicity. We propose that the use of calmodulin inhibitors in thermochemotherapy is worthy of further evaluation.

Antitumour activity of chlorimidazine on transplantable mouse tumour systems.

The antitumour activity of the newly synthesized compound, chlorimidazine (1-chlorethyl-2-chlormethyl-2-imidazoline hydrochloride), has been investigated on mouse tumour systems including L 1210 and P 388 leukaemias and Lewis lung carcinoma. It has been found that optimum treatment schedules resulted in 153% T/C in mice with L 1210 leukaemia, 159% T/C in mice bearing P 388 leukaemia and 68% TWI for Lewis lung carcinoma. Due to the results obtained the compound acts as a novel potential antitumour agent that warrants further evaluation.

Testing of 2,3-dihydro-1,3-6H-oxazine-dione (3-oxauracil) in mouse L1210 leukemia and 37 sarcoma.

The antimetabolite 2,3-dihydro-1,3-6H-oxazine-dione (3-oxauracil) has been investigated in an attempt to elucidate its effects on survival time of DBA/2 and B6D2F1 mice with L1210 leukemia and of outbred albino mice with 37 sarcoma. A significant increase in survival time was observed in mice with 37 sarcoma and slight effects were observed in mice with L1210 leukemia when treated with 3 x 12.5 or 3 x 25 mg/kg of 3-oxauracil. No toxic side effects were observed when large therapeutic doses of the drug were given.