Effect of salinity on the toxicity of diclofenac, ibuprofen and naproxen toward cyanobacterium Synechocystis salina.

Aquatic species are continuously exposed to pharmaceuticals and changeable water conditions simultaneously, which can induce changes in the toxicity of pollutants. Cyanobacterium are an organism for which less ecotoxicological tests have been performed compared to green algae. In this study, we decided to check how selected non-steroidal anti-inflammatory drugs (NSAID) affect the grow of Synechocystis salina, picocyanobacterium isolated from the Baltic Sea, with salinity as potential modulator of toxicity. S. salina was exposed to diclofenac (DCF), ibuprofen (IBF) and naproxen (NPX) (nominal 100 mg L-1) in BG11 medium and sea salt supplemented BG11 medium (38 PSU) over 96 h in continuous light at 23 °C. No acute toxicity was found in both tested salinity levels. The comparable grow rate in exposed culture compared to control culture over 4 days indicate lack of stress for several generations which need to be overcome with substantial energy consumption. S. salina was found to be halotolerant and can be species for ecotoxicology test where salinity in an additional stressor. Furthermore, resistant of S. salina to target NSAIDs provide a competitive advantage over other phytoplankton species.

Selenium Ameliorates Ibuprofen Induced Testicular Toxicity by Redox Regulation: Running Head: Se protects against NSAID induced testicular toxicity.

Despite the Cox inhibitory anti-inflammatory and antipyretic effects of most widely used non-steroidal anti-inflammatory drugs (NSAIDs), such as Ibuprofen, their chronic use is associated with a plethora of patho-physiological insults. One such toxic effect on testicular tissues is not well studied and the underlying molecular mechanisms remain unexplored. Thus, the current study is designed to evaluate the antioxidant properties of essential trace element selenium (Se) to ameliorative Ibuprofen associated testicular toxic effects. Adult male Wistar rats were divided into 3 groups and fed on diets containing different concentrations of sodium selenite, viz. 0.01 mg/kg (Se- deficient), 0.2 mg/kg (Se-adequate), or 0.5 mg/kg (Se- supplemented) for 8 weeks. After diet feeding schedule, each group was divided into two subgroups i.e., with or without the treatment of Ibuprofen (120 mg/kg Bw). The protective effect of Se was evaluated by measuring testicular Se and selenoproteins status, spermatogenic markers, histopathology and testicular redox status. Ibuprofen diminished seminal volume, sperm count, sperm motility, which correlated well increased testicular reactive oxygen species. Se deficiency exacerbated these detrimental effects of ibuprofen by increasing oxidative stress. Alternatively, Se supplementation through antioxidant enzymes mediated protective effects. Se as essential antioxidant selenoproteins ameliorates Ibuprofen induced male reproductive toxicity.

Assessing the effect of human pharmaceuticals (carbamazepine, diclofenac and ibuprofen) on the marine clam Ruditapes philippinarum: An integrative and multibiomarker approach.

The presence of pharmaceuticals in the aquatic ecosystem has become a topic of growing interest in recent years. In this study, the marine clam Ruditapes philippinarum was exposed during 14 days to concentrations close to those found in the environment: (15 µg L-1) of carbamazepine (CBZ), diclofenac (DCF) and ibuprofen (IBU), three pharmaceuticals widely used worldwide and commonly found within the aquatic environment. Additionally, exposure was followed by a depuration phase (7 days). A battery of biomarkers (superoxide dismutase SOD, catalase CAT, glutathione reductase GR, total glutathione peroxidase T-GPx, glutathione transferase GST, lipid peroxidation LPO, acetylcholinesterase AChE and metallothionein MT) was evaluated throughout the exposure and depuration. The Integrated Biomarker Response index was calculated with all selected biomarkers and used as a complementary tool in the evaluation of the organisms' health status. Exposure induced changes in the clams' biochemical responses that led to the hypothesis of the harmful role of the pharmaceuticals resulting in negative effects (changes in enzyme activities, LPO and MT levels, related to ROS production) particularly after short-term exposure. However, the clams showed the ability to cope with these imbalances by recovering their general oxidative status by the end of exposure.

Evaluación toxicológica de soluciones acuosas de ibuprofeno mediante bioensayos con Artemia salina, Allium schoenoprasum L y Lactuca sativa / Toxicological evaluation of aqueous solutions of ibuprofen through bioassays with Artemia salina, Allium schoenoprasum L y Lactuca sativa

Los fármacos son considerados dentro de los llamados contaminantes emergentes, por lo que es importante investigar los efectos tóxicos que pueden producir en el medio ambiente. Este estudio tuvo como objetivos determinar el efecto tóxico de soluciones acuosas de ibuprofeno sobre un organismo acuático Artemia salina y las semillas de Allium schoenoprasum L y Lactuca sativa. Los ensayos de toxicidad se llevaron a cabo exponiendo a los organismos de prueba a concentraciones de soluciones acuosas de ibuprofeno de 20, 10, 5, 2, 1 y 0.5 mgL-1. Se determinó el porcentaje de mortalidad para la Artemia salina y la elongación de la radícula e hipocótilo para las semillas. El porcentaje de mortalidad más elevado se presentó a la concentración de 20 mgL-1, la CL50 calculada fue de 17.386 mgL-1. Un efecto tóxico bajo se presentó en la germinación de las semillas de Allium schoenoprasum L. Se produjo la inhibición en la elongación de la radícula e hipocótilo en semillas de Allium schoenoprasum L y un efecto de estimulación en la elongación de la radícula e hipocótilo de las semillas de Lactuca sativa. El mayor efecto de inhibición se presentó a 1 y 20 mgL-1 y la mayor estimulación a 20 mgL-1

Drugs are considered within the so-called emerging pollutants, so it is important to investigate the toxic effects they can produce in the environment. The aim of this study was to determine the toxic effect of aqueous solutions of ibuprofen on an aquatic organism Artemia salina and the seeds of Allium schoenoprasum L and Lactuca sativa. The toxicity tests were carried out by exposing the test organisms to concentrations of aqueous ibuprofen solutions of 20, 10, 5, 2, 1 and 0.5 mgL-1. The percentage of mortality for Artemia salina and the elongation of the radicle and hypocotyl for the seeds were determined. The highest mortality percentage occurred at the concentration of 20 mgL-1, the calculated LC50 was 17.386 mgL-1. A low toxic effect was present in the germination of the seeds of Allium schoenoprasum L. There was inhibition in the elongation of the radicle and hypocotyl in seeds of Allium schoenoprasum L and a stimulation effect in the elongation of the radicle and hypocotyl of the seeds of Lactuca sativa. The highest inhibition effect was presented at 1 and 20mgL-1 and the highest stimulation at 20 mgL-1

Delivery of ibuprofen by natural macroporous sporopollenin exine capsules extracted from Phoenix dactylifera L.

Sporopollenin macroporous capsules (SMCs) were extracted from date palm (Phoenix dactylifera L.) spores and coated by a natural polymer composite (chitosan with glutaraldehyde). The polymer coated macroporous capsules SMC@poly were used in the in vitro-controlled delivery of ibuprofen. The materials obtained were characterized through spectral, thermal, scanning electron microscopy (SEM), X-ray diffraction (XRD), and nitrogen adsorption-desorption isotherms. The IBU loading and releasing was studied by investigating the changes in various factors such as pH, temperature, and initial concentration. The results revealed that the loading of IBU increased when the concentration of IBU was decreased, following the Langmuir adsorption isotherm. The maximum loading of the IBU was observed at pH6.0 (97.2%, with 50mg/mL). The releasing results indicate that IBU was released faster when the pH was changed from 1.4 to 7.4. In addition, the cytotoxicity of the SMC, SMC@poly, and SMC@poly-IBU were tested against human intestinal Caco-2 cell line using MTT assay, and the results revea'led that all the materials in this study were biocompatible.

Zizyphus jujuba protects against ibuprofen-induced nephrotoxicity in rats.

CONTEXT: Zizyphus jujuba Mill. (Rhamnaceae) has long been used for the treatment of anxiety and insomnia in Chinese traditional medicine. The edible part is the fruit. Different parts of Z. jujuba possess medicinal properties such as anti-inflammatory, anticancer and antifertility. OBJECTIVES: This study evaluated the therapeutic effect of Z. jujuba fruit aqueous extract (ZE) on nephrotoxicity induced by ibuprofen (IBP) in rats. MATERIALS AND METHODS: Male Sprague-Dawley rats were grouped as normal saline (control), ZE (500 mg/kg), IBP (400 mg/kg) and ZE + IBP-treated groups. After five days of oral administration, rats were sacrificed. The protective effect of ZE was evaluated by measuring kidney biomarkers, and histopathological changes of kidney were observed. Kidney antioxidant enzymes such as superoxide dismutase, catalase (CAT), glutathione S-transferase (GST) and lipid peroxidase were investigated. RESULTS: Administration of IBP resulted in a significant increase in urea and creatinine (p < 0.05) and a significant decrease in albumin and total protein (p < 0.05). Damage in glomeruli and proximal convoluted tubules was observed. IBP also increased CAT (p < 0.05) and GST (p < 0.001) activities compared to the control group. Administration of ZE with IBP significantly decreased serum urea and creatinine (p < 0.05) and reduced the severity of kidney damage. There was also a significant increase in the activities of CAT (p < 0.05) and GST (p < 0.001). DISCUSSION AND CONCLUSION: These results indicated that Z. jujuba aqueous extract could have a therapeutic role in reducing nephrotoxicity induced by ibuprofen.

Nitric oxide release is not required to decrease the ulcerogenic profile of nonsteroidal anti-inflammatory drugs.

The objective of this work was to evaluate the biological properties of a new series of nitric oxide-releasing nonsteroidal anti-inflammatory drugs (NO-NSAIDs) possessing a tyrosol linker between the NSAID and the NO-releasing moiety (PROLI/NO); however, initial screening of ester intermediates without the PROLI/NO group showed the required (desirable) efficacy/safety ratio, which questioned the need for NO in the design. In this regard, NSAID ester intermediates were potent and selective COX-2 inhibitors in vitro, showed equipotent anti-inflammatory activity compared to the corresponding parent NSAID, but showed a markedly reduced gastric toxicity when administered orally. These results provide complementary evidence to challenge the currently accepted notion that hybrid NO-NSAIDs exert their cytoprotective effects by releasing NO. Results obtained in this work constitute a good body of evidence to initiate a debate about the future replacement of NSAID prodrugs for unprotected NSAIDs (possessing a free carboxylic acid group) currently in clinical use.

The novel phospho-non-steroidal anti-inflammatory drugs, OXT-328, MDC-22 and MDC-917, inhibit adjuvant-induced arthritis in rats.

BACKGROUND AND PURPOSE: The use of non-steroidal anti-inflammatory drugs (NSAIDs) in the treatment of rheumatoid arthritis (RA) is limited by their toxicity. We evaluated the anti-inflammatory efficacy and safety of three novel modified NSAIDs, phospho-aspirin, phospho-ibuprofen and phospho-sulindac. EXPERIMENTAL APPROACH: We determined the anti-inflammatory effects and gastrointestinal safety of the phospho-NSAIDs in the rat adjuvant arthritis model and studied their mechanism of action in cultured cells, Cytokines were measured with elisa and activation of nuclear factor-κB (NF-κB) by immunohistochemistry. KEY RESULTS: All three phospho-NSAIDs showed less gastrointestinal toxicity than their parent compounds and demonstrated strong anti-inflammatory effects, essentially reversing joint inflammation and oedema. They have a broad but not uniform effect on the expression of relevant cytokines, in general decreasing IL-6 and IL-1ß and increasing IL-10 levels in rat plasma and cultured cells. Phospho-sulindac and phospho-ibuprofen but not phospho-aspirin suppressed PGE(2) production in vitro, whereas phospho-aspirin (in contrast to aspirin) showed the same effect in vivo. In joint tissues, phospho-aspirin inhibited NF-κB activation, and suppressed inflammation and bone resorption. Phospho-aspirin also inhibited Jurkat T cell proliferation. In general, phospho-aspirin had greater efficacy but different effects upon inflammatory mediators compared with aspirin. The chemical modification of the parent NSAIDs seems crucial for their safety and efficacy. CONCLUSIONS AND IMPLICATIONS: Phospho-aspirin, phospho-ibuprofen and phospho-sulindac were safer than their parent NSAIDs, were highly effective in rat adjuvant arthritis and inhibited many key mediators in the pathophysiology of RA. These novel compounds are promising candidate drugs for the treatment of RA and merit further evaluation.

Antiulcerogenic activity of bark extract of Tabebuia avellanedae, Lorentz ex Griseb.

Tabebuia avellanedae is commonly used for the treatment of peptic ulcers. We carried out this study with the ethanolic extract of bark from Tabebuia avellanedae (EET) (30-1000 mg/kg) to determine its gastroprotective activity and to clarify the pathways involved in this effect. Acute gastric ulceration in rats was produced by oral administration of ethanol and ibuprofen. After ethanol administration, the gastric wall mucus was examined. Chronic gastric ulceration was produced by injection of acetic acid in rat gastric subserosa. Anti-secretory studies were undertaken using Shay rat pylorus ligature technique and measurement of enzymatic activity of H+, K+-ATPase in vitro. Administration of EET p.o. or i.p. significantly inhibited gastric mucosa damage induced by ethanol and ibuprofen. The anti-ulcer effect was further confirmed by enhanced gastric mucus production. In pylorus ligature rats, EET significantly reduced the basal gastric acid secretion and total acidity; moreover, it inhibited the increase in total acidity induced by histamine. In addition, EET reduced the activity of H+, K+, ATPase. The results obtained in the present pharmacological assay indicate that this plant has a protective action against gastric lesions, involving the maintenance of protective factors, such as mucus and prostaglandin, besides the reduction of gastric total acidity.

Two drug interaction studies evaluating the pharmacokinetics and toxicity of pemetrexed when coadministered with aspirin or Ibuprofen in patients with advanced cancer.

PURPOSE: Pemetrexed is an antimetabolite that is structurally similar to methotrexate. Because nonsteroidal anti-inflammatory drugs (NSAID) impair methotrexate clearance and increase its toxicity, we evaluated the pharmacokinetics and toxicity of pemetrexed when coadministered with aspirin or ibuprofen in advanced cancer patients. EXPERIMENTAL DESIGN: In two independent, randomized, crossover drug interaction studies, cancer patients with a creatinine clearance (CrCl) > or =60 mL/min received an NSAID (aspirin or ibuprofen) with either the first or the second dose of pemetrexed (cycle 1 or 2). Pemetrexed (500 mg/m(2)) was infused i.v. on day 1 of a 21-day cycle, and all patients were supplemented with oral folic acid and i.m. vitamin B(12). Aspirin (325 mg) or ibuprofen (400 mg; 2 x 200 mg) was given orally every 6 hours, starting 2 days before pemetrexed administration, with the ninth and final dose taken 1 hour before infusion. Pemetrexed pharmacokinetics with and without concomitant NSAID treatment were compared for cycles 1 and 2. RESULTS: Data from 27 patients in each study were evaluable for the analysis of pemetrexed pharmacokinetics. Coadministration of aspirin did not alter pemetrexed pharmacokinetics; however, ibuprofen coadministration was associated with a 16% reduction in clearance, a 15% increase in maximum plasma concentration, and a 20% increase in area under the plasma concentration versus time curve but no significant change in V(ss) compared with pemetrexed alone. No febrile neutropenia occurred in any patient, and no increase in pemetrexed-related toxicity was associated with NSAID administration. CONCLUSIONS: Pemetrexed (500 mg/m(2)) with vitamin supplementation is well tolerated and requires no dosage adjustment when coadministered with aspirin (in patients with CrCl > or =60 mL/min) or ibuprofen (in patients with CrCl > or =80 mL/min).

Antiinflammatory and antiulcer activities of phytic acid in rats.

Maximum antiinflammatory activity of phytic acid (PA) was seen at an oral dose of 150 mg/kg in the carrageenan induced rat paw edema model. Although PA showed ability to prevent denaturation of proteins, it showed less antiinflammatory activity than ibuprofen. Ability of PA to bring down thermal denaturation of proteins might be a contributing factor in the mechanism of action against inflammation. PA, at all the doses tested, showed significant protection from ulcers induced by ibuprofen, ethanol and cold stress, with a maximum activity at 150 mg/kg. There was a significant increase in gastric tissue malondialdehyde levels in ethanol treated rats but these levels decreased following PA pretreatment. Moreover, pretreatment with PA significantly inhibited various effects of ethanol on gastric mucosa, such as, reduction in the concentration of nonprotein sulfhydryl groups, necrosis, erosions, congestion and hemorrhage. These results suggested that gastro-protective effect of PA could be mediated by its antioxidant activity and cytoprotection of gastric mucosa.

Ursodeoxycholic acid ameliorates ibuprofen-induced enteropathy in the rat.

BACKGROUND: The objective of the study was to determine whether ursodeoxycholic acid (Ursodiol) is protective against ibuprofen (IBU)-induced enteropathy. METHODS: Using the chronically catheterized rat model, IBU (60 mg/kg body weight per day) was infused via the gastric catheter twice daily. Pancreatic enzyme (PE; 10,000 U lipase/kg body weight per day) and Ursodiol (10 mg/kg body weight per day) in two doses were infused via the duodenal catheter. Rats were assigned to one of six treatment groups and were administered treatment for 20 days: control, IBU, PE, IBU + PE, IBU + Ursodiol, and IBU + PE + Ursodiol. The entire jejunum, ileum, cecum, and colon were available for histologic analysis using previously described techniques. RESULTS: Addition of Ursodiol to high-dose IBU and normal doses of PE showed a significant reduction in the percentage of rats with ulcers (P < 0.05), total number of serositis events (P < 0.01), total number of severe ulcers (P < 0.001), and an absence of ulcers in the large intestine. CONCLUSIONS: Ursodiol, the drug of choice for the treatment of cystic fibrosis liver disease, may offer a safe method of using high-dose IBU in these patients by ameliorating the enteropathy.

The effects of high-dose ibuprofen and pancreatic enzymes on the intestine of the rat.

BACKGROUND: High-dose ibuprofen therapy limits the progression of lung disease in patients with cystic fibrosis. However, ibuprofen increases intestinal permeability, which potentiates intestinal damage caused by high-dose pancreatic enzyme treatment, as was shown in a previous study by this group. In the present study, the combined effects of ibuprofen and pancreatic enzyme treatment on the intestine and liver were examined. METHODS: Using a chronically catheterized rat model, high-dose ibuprofen (60 mg/kg x day in two doses), with or without pancreatic enzyme treatment was infused into gastric and duodenal catheters, respectively, for 20 days. Six groups were studied: control group; ibuprofen treatment alone; pancreatic enzyme treatment alone (two groups: normal dose, 10,000 U lipase/kg x day and high dose, 40,000 U lipase/kg x day); and ibuprofen combined with pancreatic enzyme (two groups: ibuprofen with high-dose pancreatic enzyme and ibuprofen and low-dose pancreatic enzyme). After treatment, rats were autopsied, and complete histologic analyses of the entire intestine and liver were performed. RESULTS: Ibuprofen caused mild ulceration of the small intestine in 50% of rats. Pancreatic enzyme treatment alone did not induce ulceration of the intestine. The combination of pancreatic enzyme and ibuprofen treatment increased the severity of the ulcers in the small intestine but not the number of ulcers or the percentage of rats affected. Ibuprofen treatment alone did not cause ulcers in the large intestine, but with the addition of pancreatic enzymes, ulceration and fibrosis were present. CONCLUSIONS: Ibuprofen at doses used to limit progression of cystic fibrosis lung disease caused enteropathy in 50% of rats. There was synergism between ibuprofen and pancreatic enzyme treatment in the production of severe ulcers. Ulcers in the cecum and colon were increased with combined ibuprofen and pancreatic enzyme treatment compared with incidence in control animals.

Inhibition of arachidonate 5-lipoxygenase triggers massive apoptosis in human prostate cancer cells.

Diets high in fat are associated with an increased risk of prostate cancer, although the molecular mechanism is still unknown. We have previously reported that arachidonic acid, an omega-6 fatty acid common in the Western diet, stimulates proliferation of prostate cancer cells through production of the 5-lipoxygenase metabolite, 5-HETE (5-hydroxyeicosatetraenoic acid). We now show that 5-HETE is also a potent survival factor for human prostate cancer cells. These cells constitutively produce 5-HETE in serum-free medium with no added stimulus. Exogenous arachidonate markedly increases the production of 5-HETE. Inhibition of 5-lipoxygenase by MK886 completely blocks 5-HETE production and induces massive apoptosis in both hormone-responsive (LNCaP) and -nonresponsive (PC3) human prostate cancer cells. This cell death is very rapid: cells treated with MK886 showed mitochondrial permeability transition between 30 and 60 min, externalization of phosphatidylserine within 2 hr, and degradation of DNA to nucleosomal subunits beginning within 2-4 hr posttreatment. Cell death was effectively blocked by the thiol antioxidant, N-acetyl-L-cysteine, but not by androgen, a powerful survival factor for prostate cancer cells. Apoptosis was specific for 5-lipoxygenase-programmed cell death was not observed with inhibitors of 12-lipoxygenase, cyclooxygenase, or cytochrome P450 pathways of arachidonic acid metabolism. Exogenous 5-HETE protects these cells from apoptosis induced by 5-lipoxygenase inhibitors, confirming a critical role of 5-lipoxygenase activity in the survival of these cells. These findings provide a possible molecular mechanism by which dietary fat may influence the progression of prostate cancer.

Evaluation of the rat embryo culture system as a predictive test for human teratogens.

Ingestion of the anticonvulsant drug valproic acid and of the angiotensin converting enzyme inhibitor captopril during pregnancy has been associated with abnormal fetal outcome in humans. In contrast, the use of the antiinflammatory drug ibuprofen and the antihistamine diphenhydramine has not been documented to be embryotoxic in humans. We evaluated the rat embryo culture system as a predictive model of teratogenesis, using these four drugs as test agents. Valproic acid, ibuprofen, and diphenhydramine were embryotoxic, inducing concentration-dependent decreases in growth and a significant increase in anomalies. Valproic acid caused an increase in neural tube defects, ibuprofen increased the incidence of abnormal maxillary processes, and diphenhydramine increased the number of embryos with distorted body morphology. These abnormalities were induced at concentrations of valproic acid and diphenhydramine that are used clinically, but ibuprofen only induced toxicity at concentrations greatly exceeding the therapeutic range. Captopril was not embryotoxic up to 5 mM, the highest concentration tested. These results suggest that the rat embryo culture system produces both false positive and false negative data on the teratogenic potential of drugs. Although such an in vitro assay may be suitable to determine the mechanism of teratogenesis, it is not a sensitive indicator of potential human teratogens on its own. These data support the view that in vitro systems can only supplement clinical and epidemiological observations in humans, possibly as a method to determine mechanisms of actions of teratogens.

A study of the relative hepatotoxicity in vitro of the non-steroidal anti-inflammatory drugs ibuprofen, flurbiprofen and butibufen.

1. The cytotoxic and metabolic effects of ibuprofen, flurbiprofen and butibufen have been studied in primary cultured hepatocytes. Toxic effects were observed for all three drugs at 10 times the therapeutic plasma concentration. 2. None of the drugs affected cell survival after 48 h of continuous exposure at their therapeutic plasma concentration, although significant increases of LDH leakage were detected. 3. Ibuprofen and butibufen were the most active in impairing gluconeogenesis from lactate (88% and 76% inhibition respectively) after 6 h exposure at therapeutic plasma concentrations. 4. At 5 times therapeutic plasma concentrations, albumin synthesis was inhibited 40% (ibuprofen), 35% (flurbiprofen) and 100% (butibufen) after 6 h exposure and significant effects were also observed after 24 h exposure. 5. Urea synthesis was inhibited 11% by butibufen at its therapeutic plasma concentration but only at higher concentrations by the other drugs. 6. Butibufen was potentially the most hepatotoxic drug as it has the highest therapeutic plasma concentration and had the lowest margin between therapeutic and toxic concentrations.

[Effects of drugs on cultured cells (III)].

Toxicity of drugs cultured cell line (RK12-, EL-cell line) were exposed to various concentrations/ml medium for 48 hours and for 14 days. The morphologic changes of both cell lines observed included granulation and shrinkage of the cytoplasm, formation of long and narrow cytoplasmic projection, and appearance of giant like cells. ID50 values of ibuprofen, naproxen, Y-5544, diclofenac and aminopyrine to RK13 cell were found to be 130, 290, 200, 140 and greater than 500 mug/ml respectively. On the other hand, ID50 values of each drug to FL-cell were found to be 105, greater than 500, 180, 170 and less than 500 mug/ml respectively. Minimum concentration, cuased by the detachment of the cell from the vessel wall, was as follows: to RK13 were ibuprofen 125 approximately 250, paproxen 250 approximately 500, Y-5554 125 approximately 250, diclofenac 62.5 approximately 125, aminopyrine 1000 approximately 2000 each mug/ml. On the other hand, to FL-cell were ibuprofen 62.5 approximately 125, naproxen 125 approximately 250, Y-5554 62.5 approximately 125, DIClofenac 62.5 approximately and aminopyrine 1000 approximately 2000 each mug/ml respectively.

[Effects of drugs on cell culture (II)].

The toxicity of drugs was determined using embryonic skin and muscle from humans (Flow 1,000). The dosage causing a 50% inhibition culture growth (ID50) and minimum concentration, caused by the detachment of the cell from the vessel wall, were determined. ID50 values of ibuprofen, naproxen, Y-5554, dichlofenac and aminopyrine were found to be 150, 320,220, 110 and greater 500 mug/ml respectively. Minimum concentration, caused by the detachment of the from the vessel wall, was as follows: ibuprofen 250, naproxen 250-500, Y-5554 250-500, dichlofenac 62.50-125 and aminopyrine 1,000-2,000 mug/ml respectively.