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Ženke Wistar štakora, 200g, randomizirano podijeljene, retrobulbarno su tretirane sa: L-NAME (0,1ml/0,2mg/oko) i BPC 157 (0,1ml/0,2μg/oko, 20 minuta nakon primjene L-NAME); L-NAME (0,1ml/0,2mg/oko) i BPC 157 (0,1ml/0,2ng/oko, 20 minuta nakon primjene L-NAME); L-NAME (0,1ml/0,2mg/oko) i 0,9% NaCl (0,1ml/oko, 20 minuta nakon primjene L-NAME). Kontrolne su životinje istovremeno retrobulbarno primale jednaki volumen 0,9% NaCl. USB mikroskopskom kamerom „Veho discovery VMS-004 deluxe“ i „VOLK“ digital wide field lupom za indirektnu oftalmolskopiju snimana je očna pozadina. Inicijalno, životinje su snimane: prije retrobulbarne aplikacije L-NAME (odnosno 0,9% NaCl); 20 minuta nakon aplikacije L-NAME (odnosno 0,9% NaCl); ponovno nakon aplikacije BPC 157 (odnosno 0,9% NaCl) te nakon 20 minuta. Nadalje snimane su 1. i 2. dan nakon aplikacije te nakon 1., 2. i 4. tjedna (neposredno prije žrtvovanja). Očna je pozadina analizirana računalnim programom za analizu fotografija te patohistološkom analizom bulbusa na standardnim rezovima, uklopljenog u parafin i bojenog hemalaun eozinom. Snimano ponašanje životinja uključivalo je vrijeme imobilnosti životinja nakon stavljanja na nepoznatu podlogu i reakciju životinja na podražaj plavo-zelenom rukavicom. Retrobulbamo aplicirana L-NAME uzrokovala je vazokonstrikciju arterija mrežnice, ishemiju mrežnice te značajnu imobilnost životinja. Retrobulbarna primjena BPC 157 dovela je do oporavka mrežnične ishemije te je spriječila njenu atrofiju. Ponašanje životinja bilo je normalno., Female Wistar rats, 200 g, randomly assigned, were treated retrobulbary with: L-NAME (0.1ml/0.2mg/eye) and BPC 157 (0.1ml/0.2μg/eye 20 minutes after L-NAME application); L-NAME (0.1ml/0.2mg/eye) and BPC 157 (0.1ml/0.2ng/eye 20 minutes after L-NAME); L-NAME (0.1ml/0.2 mg/eye) and 0.9% NaCl (0.1ml/eye 20 minutes after L-NAME application). Control animals simultaneously received retrobulbar equivolume of 0.9% NaCl. With a USB microscope camera „Veho discovery VMS-004 deluxe“ and "VOLK" digital wide field loupe for indirect ophthalmoscope eye fundus was recorded in the following time periods: before retrobulbar application of L-NAME (or 0.9% NaCl); 20 minutes after L-NAME (or 0.9% NaCl); again after BPC 157 (or 0.9% NaCl), and 20 minutes after. Furthermore, recording was performed at the post-application day 1, 2, and 7, 14, 28 (last day, just before the sacrifice). The fundus was analysed by photo analysis software and by histopathologic eyeball analysis on standard sections, paraffin embedded and hemalaun-eosin stained. The animals behavior was filmed, observing the following parameters: time of immobility and reaction time to visual stimuli (blue-green glove). The retrobulbar application of L-NAME causes the vasoconstriction of the retina's arteries, retinal ischemia, while retrobulbar application of BPC 157 leads to retinal ischemia recovery, prevents its atrophy and restores normal animal behavior.

Previously, in a short term study, stable gastric pentadecapeptide BPC 157 initially counteracts increased intraocular pressure (IOP) in rats underwent episcleral cauterization (The FASEB Journal, Vo. 30 No.1 supplement 1201.4). Now, in a six month study, rats underwent episcleral cauterization instead continuously exhibiting increased IOP, showed normal intraocular pressure with BPC 157 therapy, given daily, in the whole six month period. Materials and methods Male Albino Wistar rats, 200g, anesthetized (Ketmine- HCl 50–60mg/kg + Xylazine-HCl 5–10mg/kg, Tetrakain 0.5% drops), randomly divided were used for surgery (two dorsal episcleral veins and one temporal episcleral vein isolated from the surrounding tissues ; a cautery specifically applied to the selected vein). Now, medication (pentadecapeptide BPC 157 (10μg/kg) (Diagen, Slovenia) intraperitoneally) or an equivolume of 0.9%NaCl (5ml/kg) intraperitoneally (controls)) was immediately after surgery, and then once time daily. Non-invasive IOP measurements (% of normal IOP) (after local application of Tetrakain 0.5%, by aplanation tonometer Tonopen XL always at 9.00 a.m.), and papilla nervi optici were assessed throughout 3 or 6 months after procedure. Vascularization of the eye fundus and presentation of papilla n. optici were analyzed with microcamera (Veho USB Discovery VMS-004) and Digital Widefield lupe 90D. We focused on the vessels extending into and out of the optic disc, retinal changes alterations in vessel caliber and tortuosity, optic disc pallor and leakage of the retinal arterioles and venulas, scored (1–3), as described before (The FASEB Journal, Vo. 30 No.1 supplement 1201.4). Results At 3 months after surgery controls exhibited increased values (IOP=134±5%) while BPC 157 remained values close to normal IOP=104±4%. At 6 months after surgery control rats maintained the increased values IOP=133±6%. BPC 157 exhibited normal IOP=106±5%. All BPC 157-rats exhibited only slight generalized vessel caliber irregularity (difference in diameter between arteries and veins) with a vaguely atrophic optic disc and an area not bigger than 1 optic disc diameter containing leaking of retinal vessels (Min/Med/Max 1/1/1) (Figure 1). Controls exhibited increased disturbances: either moderate generalized vessel caliber irregularity (bigger difference in diameter between arteries and veins with moderate overfilling of veins) Min/Med/Max 2/2/3 ; with moderate atrophic optic discs Min/Med/Max 2/2/3 ; and an area not more than 3 optic disc diameters containing leaking of retinal vessels Min/Med/Max 2/2/3 ; or severe generalized vessel caliber irregularity (very big difference in diameter between arteries and veins with severe overfilling of veins) with severe atrophic optic disc and an area more than 3 optic disc diameters with leaking of retinal vessels (all Min/Med/Max 3/3/3) (Figure 2). Microscopy analysis correlates well with ophthalmoscopy findings (i.e., BPC 157 groups diameter of the optic nerve remained close to normal unlike control's atrophy/thinning of the optic nerve). Conclusion Pentadecapeptide BPC 157 continuously counteracts the effects of episcleral vein cauterization.

In rats, retrobulbar L-NAME injection produces quickly severe generalized vessel caliber irregularity, leading to moderate to severe atrophic optic disc, poor presentation of the choroidal blood vessels ; retinal ischemia and atrophy, and immobile animal behavior. We hypothesize the stable gastric pentadecapeptide BPC 157 as a therapy. BPC 157 already cures ocular tissues. BPC 157 eye drops (ug- or ng-regimen) rescued total debridement of corneal epithelium (Coll Antropol 2005 ; 29(1):321–5), successfully closed perforating corneal incisions in rats (Exp Eye Res 2015 ; 136:9–15), counteracted particular NO-specific parallel miotic effect of L-NAME and L-arginine and atropine-induced mydriasis in rats and guinea pigs (Eur J Pharmacol 2016 ; 771:211– 9). In addition, BPC 157/NO-relationship is established in various experimental models and species (Curr Pharm Des 2014 ; 20(7):1126–35), providing that it would counteract the damaging effects of retrobulbar NOS-blockade and L-NAME application. Materials and methods Female Wistar rats, 200g, randomly assigned, were treated retrobulbary with: L-NAME (0.1ml/0.2mg/eye) and (a) BPC 157 (0.1ml/0.2μg/eye 20 minutes after L- NAME application) ; L-NAME (0.1ml/0.2mg/eye) and BPC 157 (0.1ml/0.2ng/eye 20 minutes after L-NAME) ; L-NAME (0.1ml/0.2 mg/eye) and 0.9% NaCl (0.1ml/eye 20 minutes after L-NAME appilication), or altenatively, (b) BPC 157 (0.1ml/0.2μg/eye 48 hours after L-NAME application) ; or 0.9% NaCl (0.1ml/eye 48 hours after L-NAME appilication). Control (sham) animals simultaneously received retrobulbar equivolume of 0.9% NaCl. With a USB microscope camera “Veho discovery VMS- 004 deluxe” and “VOLK” digital wide field loupe for indirect ophthalmoscopy eye fundus was recorded in the following time periods: before retrobulbar application of L-NAME (or 0.9% NaCl) ; 20 minutes after L-NAME (or 0.9% NaCl) ; again after BPC 157 (or 0.9% NaCl), and 20 minutes after. Furthermore, recording was performed at the post-application day 1., 2., and 7., 14., 28. (last day, just before the sacrifice). The fundus were analyzed by photo analysis software and by histopathologic eyeball analysis on standard sections, paraffin embedded and hemalaun-eosin stained. The animals behavior was filmed, observing the following parameters: time of immobility (no movement period) and reaction time to viusal stimuli (blue- green glove). Results Interestingly, given either shortly after L-NAME, or, alternatively, two days later, retrobulbar application of BPC 157 leads to retinal ischemia recovery, prevents its atrophy and restors normal animal behavior (Figure 1, Figure 2). Conclusion BPC 157 was originally an anti-ulcer peptide used in trials for ulcerative colitis and now is in trials for the treatment of multiple sclerosis that largely interacts with NO- system (Curr Pharm Des. 2014 ; 20(7):1126–35). Thereby, the noted damaging effect of retrobulbar application L-NAME in rats that was counteracted by pentadecapeptide BPC 157, provides a particular promissing base for further research and application.

Female Wistar rats, 200 g, randomly assigned, were treated retrobulbary with: L-NAME (0.1ml/0.2mg/eye) and BPC 157 (0.1ml/0.2μg/eye 20 minutes after L-NAME application); L-NAME (0.1ml/0.2mg/eye) and BPC 157 (0.1ml/0.2ng/eye 20 minutes after L-NAME); L-NAME (0.1ml/0.2 mg/eye) and 0.9% NaCl (0.1ml/eye 20 minutes after L-NAME application). Control animals simultaneously received retrobulbar equivolume of 0.9% NaCl. With a USB microscope camera „Veho discovery VMS-004 deluxe“ and "VOLK" digital wide field loupe for indirect ophthalmoscope eye fundus was recorded in the following time periods: before retrobulbar application of L-NAME (or 0.9% NaCl); 20 minutes after L-NAME (or 0.9% NaCl); again after BPC 157 (or 0.9% NaCl), and 20 minutes after. Furthermore, recording was performed at the post-application day 1, 2, and 7, 14, 28 (last day, just before the sacrifice). The fundus was analysed by photo analysis software and by histopathologic eyeball analysis on standard sections, paraffin embedded and hemalaun-eosin stained. The animals behavior was filmed, observing the following parameters: time of immobility and reaction time to visual stimuli (blue-green glove). The retrobulbar application of L-NAME causes the vasoconstriction of the retina's arteries, retinal ischemia, while retrobulbar application of BPC 157 leads to retinal ischemia recovery, prevents its atrophy and restores normal animal behavior.

In living rats, atropine-mydriasis would depend on NO-related mechanisms in a particular way: both L- NAME, a NOS-blocker and L-arginine, a NOS- substrate wouldexhibit a miotic effect, not yet described ; the atropine-mydriasis and L-NAME- and L-arginine-miosis, could consequently be counteracted by BPC 157 due to its interactions with the NO-system and sphincter function. We administraded BPC 157 (10µg, 10ng, 10pg/kg), L- NAME (5mg/kg), and/or L-arginine (100mg/kg) alone/together intraperitoneally in healthy rats (miosis-studies) or at 30 min after 1% atropine/2drops/eye (mydriasis-studies). BPC 157 does not alter normal pupil diameter while miosis occured promptly after both L-NAME and L-arginine, lasting for more than 3h, when they were given alone. When L-NAME+L-arginine were given together, this miosis was shorter, suggesting that their separate miotic effects are competitive and may antagonize each other. These L-NAME-/L-arginine- effects were both shortened in rats that received BPC 157. In case of maximal atropine-mydriasis all of the agents, BPC 157, L-NAME and L-arginine, induce a counteraction, commencing sooner in BPC 157-rats, but all of the agents lead to completely blunted atropine-mydriasis returning to regular pupil size hours sooner than in atropine-rats which received saline. Concluding, BPC 157 may counteracted those effects due to its interactions with the NO-system and sphincter function. Supported by Grant 108-1083570-3635, Croatia.

Possibly, acute esophagitis and pancreatitis cause each other, and we focused on sphincteric failure as the common causative key able to induce either esophagitis and acute pancreatitis or both of them, and thereby investigate the presence of a common therapy nominator. This may be an anti-ulcer pentadecapeptide BPC 157 (tested for inflammatory bowel disease, wound treatment) affecting esophagitis, lower esophageal and pyloric sphincters failure and acute pancreatitis (10 μg/kg, 10 ng/kg intraperitoneally or in drinking water). The esophagitis-sphincter failure procedure (i.e., insertion of the tubes into the sphincters, lower esophageal and pyloric) and acute pancreatitis procedure (i.e., bile duct ligation) were combined in rats. Esophageal manometry was done in acute pancreatitis patients. In rats acute pancreatitis procedure produced also esophagitis and both sphincter failure, decreased pressure 24 h post-surgery. Furthermore, bile duct ligation alone immediately declines the pressure in both sphincters. Vice versa, the esophagitis- sphincter failure procedure alone produced acute pancreatitis. What's more, these lesions (esophagitis, sphincter failure, acute pancreatitis when combined) aggravate each other (tubes into sphincters and ligated bile duct). Counteraction occurred by BPC 157 therapies. In acute pancreatitis patients lower pressure at rest was in both esophageal sphincters in acute pancreatitis patients. We conclude that BPC 157 could cure esophagitis/sphincter/acute pancreatitis healing failure.