Effect of polyunsaturated fatty acids and their metabolites on bleomycin-induced cytotoxic action on human neuroblastoma cells in vitro.

In the present study, we noted that bleomycin induced growth inhibitory action was augmented by all the polyunsaturated fatty acids (PUFAs) tested on human neuroblastoma IMR-32 (0.5 × 10(4) cells/100 µl of IMR) cells (EPA > DHA > ALA = GLA = AA > DGLA = LA: ∼ 60, 40, 30, 10-20% respectively) at the maximum doses used. Of all the prostaglandins (PGE1, PGE2, PGF2α, and PGI2) and leukotrienes (LTD4 and LTE4) tested; PGE1, PGE2 and LTD4 inhibited the growth of IMR-32 cells to a significant degree at the highest doses used. Lipoxin A4 (LXA4), 19,20-dihydroxydocosapentaenoate (19, 20 DiHDPA) and 10(S),17(S)-dihydroxy-4Z,7Z,11E,13Z,15E,19Z-docosahexaenoic acid (protectin: 10(S),17(S)DiHDoHE), metabolites of DHA, significantly inhibited the growth of IMR-32 cells. Pre-treatment with AA, GLA, DGLA and EPA and simultaneous treatment with all PUFAs used in the study augmented growth inhibitory action of bleomycin. Surprisingly, both indomethacin and nordihydroguaiaretic acid (NDGA) at 60 and 20 µg/ml respectively enhanced the growth of IMR-32 cells even in the presence of bleomycin. AA enhanced oxidant stress in IMR-32 cells as evidenced by an increase in lipid peroxides, superoxide dismutase levels and glutathione peroxidase activity. These results suggest that PUFAs suppress growth of human neuroblastoma cells, augment growth inhibitory action of bleomycin by enhancing formation of lipid peroxides and altering the status of anti-oxidants and, in all probability, increase the formation of lipoxins, resolvins and protectins from their respective precursors that possess growth inhibitory actions

Surgical management of urinary obstruction in young ruminants by tube cystotomy: A report of 24 cases

Objectives: To assess the efficacy of tube cystotomy along with medical dissolution of calaculi as a sole strategy for correction of obstructive urolithiasis in young ruminants. Materials and methods: Young bull calves (n=24; 10 Ongole claves and 14 Murrah buffalo calves) suffering from complete/partial urinary retention aging between 1 to 6 months were treated by tube cystotomy along with oral administration of ammonium chloride dosed at 10 gm/Kg body weight (bwt) orally for 15 days, streptopencillin dosed at 100 mg/10 Kg bwt through intramuscular (im) route for 5 days, and meloxicam dosed at 0.2 mg/Kg bwt through im route for 3 days. Results: Total 23 (95.83%) out of 24 calves started passing urine normally through the natural orifice 10-15 days postoperatively. None of the recovered animals (n=23) exhibited recurrence of symptoms, establishing the superiority of the technique in resolving the condition. Conclusion: Tube cystotomy, when performed at an early stage can prevent mortality of calves due to cystorrhexis, uroperitoneum and consequent uremia. This can avoid the painful and most tedious cysto-urethrotomy in young ruminants. [J Adv Vet Anim Res 2016; 3(2.000): 188-191

Effect of protectin on IMR-32 cells.

<p>IMR-32 cells were exposed to different doses (1, 5, 10 ng/ml) of protectin (19,20-DiHDPA, 10(S),17(S)-DiHDoHE) and incubated for 24 hrs. At the end of treatment period, cell viability was measured by MTT assay. All values are expressed as mean ± standard error (n = 6). *P<0.05 when compared to control.</p

Effect of lipoxin A4 on IMR-32 cells.

<p>IMR-32 cells were exposed to different doses (1, 5, 10 ng/ml) of lipoxin A4 and incubated for 24 hrs. At the end of treatment period, cell viability was measured by MTT assay. All values are expressed as mean ± standard error (n = 6). *P<0.05 when compared to control. LXA₄– Lipoxin A₄.</p

Effect of resolvin on IMR-32 cells.

<p>IMR-32 cells were exposed to different doses (1, 5, 10 ng/ml) of resolvin (D1, D2) and incubated for 24 hrs. At the end of treatment period, cell viability was measured by MTT assay. All values are expressed as mean ± standard error (n = 6). *P<0.05 when compared to control.</p

Effect of prostaglandin on IMR-32 cells.

<p>IMR-32 cells were exposed to different doses (10, 50, 100 ng/ml) of prostaglandins (PGE1, PGE2, PGF2α, PGI2) and incubated for 24 hrs. At the end of treatment period, cell viability was measured by MTT assay. All values are expressed as mean ± standard error (n = 6). *P<0.05 when compared to control. PG = Prostaglandin.</p

Estimation of nitric oxide and lipid peroxidation in spent media and cell lysates of IMR-32 cells.

<p>IMR-32 cells were treated with AA 30 µg/ml and bleomycin 60 µg/ml and incubated for 24 hrs. At the end of treatment period, supernatant was collected and cells were lysed. All values are expressed as mean ± standard error (n = 3). *p<0.05 when compared to control. BLM = Bleomycin, AA = Arachidonic Acid.</p><p>Estimation of nitric oxide and lipid peroxidation in spent media and cell lysates of IMR-32 cells.</p

Effect of protectin on bleomycin induced cytotoxicity in IMR-32 cells.

<p>(A) Effect of pre-treatment with different doses (1, 5, 10 ng/ml) of protectin (19,20-DiHDPA, 10(S),17(S)-DiHDoHE) on bleomycin (60 µg/ml) induced cytotoxicity to IMR-32 cells. Cells were pre incubated with protectin for 5 hrs and then bleomycin for 24 hrs. MTT assay was performed. (B) Effect of simultaneous treatment with different doses (1, 5, 10 ng/ml) of protectin (19,20-DiHDPA, 10(S),17(S)-DiHDoHE) on bleomycin (60 µg/ml) induced cytotoxicity to IMR-32 cells. Cells were pre incubated with plain media for 5 hrs and then protectin and bleomycin were added and incubated for 24 hrs. MTT assay was performed. All values are expressed as mean ± standard error (n = 6). *P<0.05 when compared to control. BLM – Bleomycin.</p

Effect of COX and LOX inhibitor on IMR-32 cells.

<p>(A) Effect of indomethacin (20, 40, 60 µg/ml) on viability of IMR-32 cells during 24 hrs incubation. (B) Effect of NDGA (5, 10, 20 µg/ml) on viability of IMR-32 cells during 24 hrs incubation. All values are expressed as mean ± standard error (n = 6). *P<0.05 when compared to control. ID = Indomethacin, NDGA = Nordihydroguariaretic acid.</p

Effect of Leukotriene on bleomycin induced cytotoxicity in IMR-32 cells.

<p>(A) Effect of pre-treatment with different doses (10, 50, 100 ng/ml) of leukotriene (LTD4, LTE4) on bleomycin (60 µg/ml) induced cytotoxicity to IMR-32 cells. Cells were pre incubated with leukotriene for 5 hrs and then bleomycin for 24 hrs. MTT assay was performed. (B) Effect of simultaneous treatment with different doses (10, 50, 100 ng/ml) of leukotriene (LTD4, LTE4) on bleomycin (60 µg/ml) induced cytotoxicity to IMR-32 cells. Cells were pre incubated with plain media for 5 hrs and then leukotriene and bleomycin were added and incubated for 24 hrs. MTT assay was performed. All values are expressed as mean ± standard error (n = 6). *P<0.05 when compared to control; #P<0.05 when compared to bleomycin. BLM – Bleomycin, LT - Leukotriene.</p