Characteristics of the acuteinflammatory reaction in macrophagemigration inhibitory factor deficientmice

Cilj ovog rada je bio da se ispitaju karakteristike sterilne zapaljenske reakcije u miševa sa delecijom Mif gena, korišćenjem modela subkutane implantacije polivinilskih sunđera. Iako je MIF poznat pro-inflamacijski citokin o njegovom uticaju na procese regeneracije nakon oštećenja tkiva postoje kontradiktorni podaci. Zbog toga su u pokušaju razumevanja ovog problema ispitivani dinamika promena ćelijskih elemenata u eksudatu rane, procesi apoptoze inflamacijskih ćelija i njihova funkcionalna sposobnost in vitro (adhezivnost i produkcija NO). Ovi parametri su povezivani sa nivoom različitih biomolekula u eksudatu (pro- i anti-inflamacijskih citokina), hemokina; faktora rasta reaktanata akutne faze i NO). Dobijeni rezultati su poređeni sa rezultatima kod kontrolnog divljeg soja C57BL/6 (Mif+/+) miševa sa očuvanim Mif genom. Pokazano je da ranu fazu inflamacije (24 časa posle implantacije sunđera) karakteriše smanjeno nakupljanje neutrofilnih granulocita, manja produkcija IL-β, IFN-γ, IL-12 i NO, povećana produkcija IL-33 i GM-CSF i neočekivano manja apoptoza neutrofila u odnosu na divlji soj miševa. Shodno očekivanju, inflamacijske ćelije u kulturi su produkovale više NO, imale veću spontanu apoptozu koja se bitnije nije menjala pod uticajem LPS-a, ali su takođe neočekivano imale povećanu adhezivnost za plastiku. Kasnu fazu inflamacije (10. dana od implantacije sunđera) je karakterisalo manje nakupljanje granulocita; makrofaga i dendritskih ćelija što nije bilo povezano sa brojem neutrofila i monocita u perifernoj krvi, veće stvaranje džinovskih multijedarnih makrofaga i povećanje apoptoze kako neutrofila tako i makrofaga. U eksudatu su nađeni niži nivoi NO, IL-β, TNF-α i CXCL10 a povećani nivoi IL-12, IL- 33, IL-1Ra i CRP. Inflamacijske ćelije su u kulturi produkovale niže nivoe NO ali su, slično kao u ranoj fazi, imale povećanu spontanu i indukovanu adhezivnost za plastiku, u odnosu na divlji soj miševa. Na osnovu dobijenih rezultata se može zaključiti da predominacija anti-inflamacijskih medijatora kod Mif-/- miševa, praćena smanjenom mobilizacijom inflamacijskih ćelija u povređeno tkivo i povećanjem njihove apoptoze, mogu delovati povoljno na bržu rezoluciju inflamacije

Characteristics of the acuteinflammatory reaction in macrophagemigration inhibitory factor deficientmice

Cilj ovog rada je bio da se ispitaju karakteristike sterilne zapaljenske reakcije u miševa sa delecijom Mif gena, korišćenjem modela subkutane implantacije polivinilskih sunđera. Iako je MIF poznat pro-inflamacijski citokin o njegovom uticaju na procese regeneracije nakon oštećenja tkiva postoje kontradiktorni podaci. Zbog toga su u pokušaju razumevanja ovog problema ispitivani dinamika promena ćelijskih elemenata u eksudatu rane, procesi apoptoze inflamacijskih ćelija i njihova funkcionalna sposobnost in vitro (adhezivnost i produkcija NO). Ovi parametri su povezivani sa nivoom različitih biomolekula u eksudatu (pro- i anti-inflamacijskih citokina), hemokina; faktora rasta reaktanata akutne faze i NO). Dobijeni rezultati su poređeni sa rezultatima kod kontrolnog divljeg soja C57BL/6 (Mif+/+) miševa sa očuvanim Mif genom. Pokazano je da ranu fazu inflamacije (24 časa posle implantacije sunđera) karakteriše smanjeno nakupljanje neutrofilnih granulocita, manja produkcija IL-β, IFN-γ, IL-12 i NO, povećana produkcija IL-33 i GM-CSF i neočekivano manja apoptoza neutrofila u odnosu na divlji soj miševa. Shodno očekivanju, inflamacijske ćelije u kulturi su produkovale više NO, imale veću spontanu apoptozu koja se bitnije nije menjala pod uticajem LPS-a, ali su takođe neočekivano imale povećanu adhezivnost za plastiku. Kasnu fazu inflamacije (10. dana od implantacije sunđera) je karakterisalo manje nakupljanje granulocita; makrofaga i dendritskih ćelija što nije bilo povezano sa brojem neutrofila i monocita u perifernoj krvi, veće stvaranje džinovskih multijedarnih makrofaga i povećanje apoptoze kako neutrofila tako i makrofaga. U eksudatu su nađeni niži nivoi NO, IL-β, TNF-α i CXCL10 a povećani nivoi IL-12, IL- 33, IL-1Ra i CRP. Inflamacijske ćelije su u kulturi produkovale niže nivoe NO ali su, slično kao u ranoj fazi, imale povećanu spontanu i indukovanu adhezivnost za plastiku, u odnosu na divlji soj miševa. Na osnovu dobijenih rezultata se može zaključiti da predominacija anti-inflamacijskih medijatora kod Mif-/- miševa, praćena smanjenom mobilizacijom inflamacijskih ćelija u povređeno tkivo i povećanjem njihove apoptoze, mogu delovati povoljno na bržu rezoluciju inflamacije

The effect of zinc diet on distribution of fatty acid in blood plasma chylomicrons

The effect of dietary zinc on the distribution of fatty acids in blood plasma chylomicrons was studied. The experiment was performed on male Mongolian Gerbils, fed ad libitum for 3 weeks with standard diet, containing 8 or 38 mg of Zn per kg of food (low zinc diet group, LZ, and saturated zinc diet group, SZ, respectivelly). At the 21st day gerbils were given sunflower oil by gavage. After 2.5 hours, blood was collected from abdominal vein, and samples were pooled (five animals per one sample). Chylomicron fractions were isolated by ultracentrifugation and mass of dried chylomicrons was measured. Fatty acid composition was analyzed by gas-liquid chromatography. Significantly higher amount of chylomicrons in blood plasma of SZ than in LZ group was found. In chylomicrons, following fatty acids were found: 16:0, 16:1, 17:0, 17:1, 18:0, 18:1, 18:2, 18:3, 20:0, 21:0 and 20:4. The amount of individual fatty acids in chylomicrons in both groups was similar, except 20:4 where lower amount in SZ group was found. Zinc diet did not affect fatty acid distribution in chylomicrons of both groups. Animals fed with zinc saturated diet had higher amount of fatty acids in blood plasma. Observed results suggest that dietary zinc influences the quantity of fatty acids absorption but not its distribution in chylomicrons.Ispitivan je efekat unosa dijetarnog cinka na distribuciju masnih kiselina u hilomikronima plazme. Eksperiment je rađen na mužjacima gerbila, hranjenim standardnom dijetom, ad libitum, u toku 3 nedelje hrana je sadržala 8 ili 38 mg cinka po kg hrane (grupa sa smanjenim unosom cinka, LZ i grupa sa povećanim unosom cinka, SZ, respektivno). Dvadeset prvog dana, životinjama je dato suncokretovo ulje sondom. Nakon 2,5 časa, krv je uzimana iz abdominalne vene i uzorci su pulovani (5 životinja za jedan uzorak). Hilomikronske frakcije su izolovane ultracentrifugiranjem i merena je masa suvih hilomikrona. Masnokiselinski sastav lipida u hilomikronima je određivan gasno-tečnom hromatografijom. Nađena je značajno veća količina hilomikrona u plazmi SZ grupe nego u LZ grupi. U hilomikronima su nađene sledeće masne kiseline: 16:0, 16:1, 17:0, 17:1, 18:0, 18:1, 18:2, 18:3, 20:0, 21:0 i 20:4. Zastupljenost pojedinačnih masnih kiselina u hilomikronima obe grupe bila je slična, izuzev za 20:4 gde je izmerena niža količina u SZ grupi. Ishrana cinkom ne utiče na distribuciju većine masnih kiselina u hilomikronima. Dobijeni rezultati ukazuju da cink utiče na metabolizam masti i to u delu formiranja hilomikrona, ali je uticaj ograničen na količinu formiranih hilomikrona, ali ne i distribuciju pojedinih masnih kiselina u hilomikronima.nul

Effects of low, adequate and high dietary zinc intake on metabolic interactions between zinc, copper and iron in different mongolian gerbil tissues

This study examined the influence of low, adequate and high dietary zinc intake on the growth and metabolic interactions between zinc, copper and iron in different Mongolian gerbil (Meriones unguiculatus) tissues. Animals were fed a basal diet (for 3 wk) containing a low-zinc level (8 mg/ kg diet, LZ-group), adequate zinc level (25 mg/kg diet, AZ-group) and saturated zinc level (38 mg/ kg diet, SZ-group). After comparing the body weight of gerbils, the present study demonstrated that the LZ-group (51.68 g +/- 3.35) showed growth retardation, contrary to the AZ- and SZ-groups (58.03 g +/- 2.18 and 62.35 g +/- 2.04, respectively). Concentrations of zinc in the liver, kidneys and testes in the LZ-group (41.28, 19.58, 22.55 mg/kg, respectively) were significantly lower than in the AZ-group (57.27, 23.73, 28.79 mg/kg, respectively) and the SZ-group (73.06, 30.07, 34.52 mg/kg, respectively). Results also showed that copper and iron concentrations in the kidney, heart and liver were significantly higher in the LZ-group (Cu 11.45, 7.01, 7.95 mg/kg; Fe 116.19, 126.07, 299.47 mg/kg) than in the AZ-group (Cu 8.72, 5.6, 6.5 mg/kg; Fe 97.27, 97.27, 250.25 mg/kg) and the SZ-group (Cu 6.48, 4.15, 4.8 mg/kg; Fe 74.95, 77.95, 200.27 mg/kg). There was an increase of testis copper concentration in the LZ-group (4.9 mg/kg) compared to the AZ- and SZ-groups (3.99 and 3.05 mg/kg, respectively), and there was no significant difference in testis iron concentration between animals in different diet groups. These results showed that a low-zinc diet had negative effects on growth and the concentration of zinc in the kidneys, liver and testes. This also affected homeostasis of copper and iron by increasing its distribution in the kidneys, liver and heart tissue. However, in testes only copper concentration was increased

Efekat ishrane cinkom na distribuciju masnih kiselina u hilomikronima plazme

The effect of dietary zinc on the distribution of fatty acids in blood plasma chylomicrons was studied. The experiment was performed on male Mongolian Gerbils, fed ad libitum for 3 weeks with standard diet, containing 8 or 38 mg of Zn per kg of food (low zinc diet group, LZ, and saturated zinc diet group, SZ, respectivelly). At the 21st day gerbils were given sunflower oil by gavage. After 2.5 hours, blood was collected from abdominal vein, and samples were pooled (five animals per one sample). Chylomicron fractions were isolated by ultracentrifugation and mass of dried chylomicrons was measured. Fatty acid composition was analyzed by gas-liquid chromatography. Significantly higher amount of chylomicrons in blood plasma of SZ than in LZ group was found. In chylomicrons, following fatty acids were found: 16:0, 16:1, 17:0, 17:1, 18:0, 18:1, 18:2, 18:3, 20:0, 21:0 and 20:4. The amount of individual fatty acids in chylomicrons in both groups was similar, except 20:4 where lower amount in SZ group was found. Zinc diet did not affect fatty acid distribution in chylomicrons of both groups. Animals fed with zinc saturated diet had higher amount of fatty acids in blood plasma. Observed results suggest that dietary zinc influences the quantity of fatty acids absorption but not its distribution in chylomicrons.Ispitivan je efekat unosa dijetarnog cinka na distribuciju masnih kiselina u hilomikronima plazme. Eksperiment je rađen na mužjacima gerbila, hranjenim standardnom dijetom, ad libitum, u toku 3 nedelje hrana je sadržala 8 ili 38 mg cinka po kg hrane (grupa sa smanjenim unosom cinka, LZ i grupa sa povećanim unosom cinka, SZ, respektivno). Dvadeset prvog dana, životinjama je dato suncokretovo ulje sondom. Nakon 2,5 časa, krv je uzimana iz abdominalne vene i uzorci su pulovani (5 životinja za jedan uzorak). Hilomikronske frakcije su izolovane ultracentrifugiranjem i merena je masa suvih hilomikrona. Masnokiselinski sastav lipida u hilomikronima je određivan gasno-tečnom hromatografijom. Nađena je značajno veća količina hilomikrona u plazmi SZ grupe nego u LZ grupi. U hilomikronima su nađene sledeće masne kiseline: 16:0, 16:1, 17:0, 17:1, 18:0, 18:1, 18:2, 18:3, 20:0, 21:0 i 20:4. Zastupljenost pojedinačnih masnih kiselina u hilomikronima obe grupe bila je slična, izuzev za 20:4 gde je izmerena niža količina u SZ grupi. Ishrana cinkom ne utiče na distribuciju većine masnih kiselina u hilomikronima. Dobijeni rezultati ukazuju da cink utiče na metabolizam masti i to u delu formiranja hilomikrona, ali je uticaj ograničen na količinu formiranih hilomikrona, ali ne i distribuciju pojedinih masnih kiselina u hilomikronima

The effect of zinc diet on distribution of fatty acid in blood plasma chylomicrons

The effect of dietary zinc on the distribution of fatty acids in blood plasma chylomicrons was studied. The experiment was performed on male Mongolian Gerbils, fed ad libitum for 3 weeks with standard diet, containing 8 or 38 mg of Zn per kg of food (low zinc diet group, LZ, and saturated zinc diet group, SZ, respectivelly). At the 21st day gerbils were given sunflower oil by gavage. After 2.5 hours, blood was collected from abdominal vein, and samples were pooled (five animals per one sample). Chylomicron fractions were isolated by ultracentrifugation and mass of dried chylomicrons was measured. Fatty acid composition was analyzed by gas-liquid chromatography. Significantly higher amount of chylomicrons in blood plasma of SZ than in LZ group was found. In chylomicrons, following fatty acids were found: 16:0, 16:1, 17:0, 17:1, 18:0, 18:1, 18:2, 18:3, 20:0, 21:0 and 20:4. The amount of individual fatty acids in chylomicrons in both groups was similar, except 20:4 where lower amount in SZ group was found. Zinc diet did not affect fatty acid distribution in chylomicrons of both groups. Animals fed with zinc saturated diet had higher amount of fatty acids in blood plasma. Observed results suggest that dietary zinc influences the quantity of fatty acids absorption but not its distribution in chylomicrons.Ispitivan je efekat unosa dijetarnog cinka na distribuciju masnih kiselina u hilomikronima plazme. Eksperiment je rađen na mužjacima gerbila, hranjenim standardnom dijetom, ad libitum, u toku 3 nedelje hrana je sadržala 8 ili 38 mg cinka po kg hrane (grupa sa smanjenim unosom cinka, LZ i grupa sa povećanim unosom cinka, SZ, respektivno). Dvadeset prvog dana, životinjama je dato suncokretovo ulje sondom. Nakon 2,5 časa, krv je uzimana iz abdominalne vene i uzorci su pulovani (5 životinja za jedan uzorak). Hilomikronske frakcije su izolovane ultracentrifugiranjem i merena je masa suvih hilomikrona. Masnokiselinski sastav lipida u hilomikronima je određivan gasno-tečnom hromatografijom. Nađena je značajno veća količina hilomikrona u plazmi SZ grupe nego u LZ grupi. U hilomikronima su nađene sledeće masne kiseline: 16:0, 16:1, 17:0, 17:1, 18:0, 18:1, 18:2, 18:3, 20:0, 21:0 i 20:4. Zastupljenost pojedinačnih masnih kiselina u hilomikronima obe grupe bila je slična, izuzev za 20:4 gde je izmerena niža količina u SZ grupi. Ishrana cinkom ne utiče na distribuciju većine masnih kiselina u hilomikronima. Dobijeni rezultati ukazuju da cink utiče na metabolizam masti i to u delu formiranja hilomikrona, ali je uticaj ograničen na količinu formiranih hilomikrona, ali ne i distribuciju pojedinih masnih kiselina u hilomikronima.nul

Vitamin B complex treatment improves motor nerve regeneration and recovery of muscle function in a rodent model of peripheral nerve injury

It is well known that the peripheral nervous system has a good potential for regeneration. The aim of this study was to explore the effects of vitamin B therapy (with a complex of vitamins B1, B2, B3, B5, B6, and B12) on motor nerve recovery after femoral nerve injury. Our study was conducted on an experimental animal model of femoral nerve injury in rats. All animals used in the experiment were subjected to the same set of analyses. A behavior test was used for the assessment of motor function recovery. Body weight was measured and electromyography was performed in order to assess recovery of quadriceps muscle. Samples of muscles and nerves were used for counting nuclei and determining nuclear density. The results of this study showed enhanced functional recovery, including improved walking, a decreased level of muscle atrophy and better electromyography recovery after administration of vitamin B complex. Further, after 14 days of treatment with the vitamin B complex nuclear nerve and muscle density was significantly lowered. In conclusion, using a model of femoral nerve injury we demonstrated that the application of vitamin B complex improved recovery of motor nerve in rats. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. 175033