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    42 Garcia et af to maximize its potential, Nanotechnology. such as nanoliposome-encapsulation, was developed io increase the bioavailability (Fang and Bhandari, 2010) of compounds such as quercetin. It aids in the protection and absorption of the drugs that are being administered. However, quercetin was found to inhibit platelet aggregation (Tzeng ef al. 1991) and was also found that high amounts of quercetin can cause kidney damage (Ehriich, 2011). With the expectedincrease in quercetin bioavailability becuse of nanolipasome- encapsulation, it is assumed that these effects will also be increased. McCullagh and Ehrman (1975) reported that dietary cholesterol can also have an effect on the kidneys of dogs. However, studies on the hematologic profile and biochemical values in adult dogs given nanoliposome-encapsulsted malunpgay phenolic extract (which contains quercetin) and cholesterol have not yet been done. Therefore, it was investigated in the current study. The resubs of this study can be used as a basis for future studies on the use of nanolipesome encapsulation in the production of nutraceuticals for dogs. MATERIALS AND METHODS: Six adultmale dogs approximately 2-3 years old with body condition score benween 4.6/9 (Laflamme, 1907) were used in this study. These dogs were collected from Los Banos, Laguna and were housed in the kennel station at the University of the Philippines’ Veterinary Teaching Hospitsl-Mashas Station. The dogs were randomly assigned to three treatments. Filler capeules containing rice bran and magnesium steate were given to Group A (contra) cogs (A £42). Non-encapsulateg malungaay chenolc erect Capsules were given to Group & dogs (1 & 82). Group C dogs (C }) received the nanoliposome- encapsulated maluny enolic extract capsules. One hundred milligram cholesterol si egay pA p a capsules were also given to all dogs. The capsules were produced and provided by ‘Smart-Functional Biomaterials Laboratory of the College of Arts and Sciences, University of the Philippines Los Bafos. The non-encapsulated malunggay phenolic exract was sroduced using the ethane! extrecion process described in the study of Oluduro (20712). he nanoliposome-ancapsulated malunggay phenobe extract was produced using thin- fim hycrstion methoa deserbec by us e! sl (2012) All dags underwent initial hematologic profiling and biochemical value testing including blood ures nitrogen (BUN), creatinine, cholesterol, triglyceride, high density lipoprotein (HDL), low density lipopratein (LDL), aspartate aminotransferase (AST) and alanine aminotransferase (ALT). The blood samples for hematolagic profiling and serum samples forthe biochemical tests were collected fram either the cephalic vein or the jugular vein and were submitted to a laboratory for processing. Hematologic: profiling inchided total white blood cell (TWEC) count, differential WEC count, packed cell volume (PCV) and platelet count ‘After the initial testing at Week 0, capsules containing 100 mg cholesterol were hen srally fo each doy every dey for ong week After one week (IVesk ‘) of ging Cholesterol, hematologic profiling and lipid profiling were repeated. With the adde cholesterol oral administration, it is assumed that dyslipidemia occurred. Then. the assigned treatment capsules were orally given along with the 100 mg cholesterol capsules for four weeks, After four weeks of treatment, all tests were repeated (Week 5). Weekly hematologic profiing and lipid profiling were done for the whole duration of the study. Routine physical examination (obtaining the heart rate, respiratory rate, body temperature and body condition score) was done twice a week. The body weight was measured at the a Garcia ef al Nakagawa K, Kawagoe M, Yoshimura M, Arata H, Minamikawa T, Nakamura M and Meteumaa 2000 Oierenta efcis af tavancid quercetin on geasine came indu rophilic ant ilic radical generators in tic omal fractions nts Tesh Sci ‘ae sO 30. ° seats lees Nelson R, Turnwald Gand Willard MD. 2004. Endocrine, Metabolic, and Lipid Disorders. In: Wllard MD aind Tvedten H (eds.) Smaif Animal Clinical Diagnosis by Laboratory Methods, 4° ed. St Louis, Missouri: Elsevier. ‘Oluduro AQ. 2012. Evaluation of antimicrobial properties and nutritional potentials of Monngaotefera Lam. leaf in South-Westem Nigeria. Asiaysian J iicrabiol £ (2): 50-87. Reinboth M, Wolfram 5, Abraham G, Ungemach F and Cermak R. 2010.0ral saagieoly of quercetin from different quercetin glycosides in dogs. Bnt J Mutr 104 (2): 198- 203. Russell KE. 2010. Platelet kinetics and laboratory evaluation of thrombocytopenia. In: Weiss DJ and Wardrop KJ (eds.). Schalm’s Veterinary Hematology. 6th ed. Ames, lowa: Wiley-Blackwell. Sachan D, Jain 5 and Singh N. 2011.In-vitro and in-vive efficacy of Jf oleifera plant contituentsinurclithiasis and antilithiatic drug. int J Phanmaceut Sci Res 27): 1838- 1644. Shoskes DA. 1998. Effect of bioflavonoids quercetin and curcumin on ischemic renal inluy Trans 68: 147-152. ‘SchultzeAE. 2010. Interpretation of canine leukocyte responses. In: Weiss DJ and Wardrop Kd (eds.). Schalm’s Veterinary fematlons @ ed. Ames, lowa: Wiley-Blackavell, Tang ¥, Chao G, Xing M, Li, Zhu L, Wang D, Yang X, Liu Z and Yao P. 2012. Quercetin prevents ethanol-induced dyslipidemia and mitochondrial oxidative damage. Food Chem Toxicol 1194-1200. ThomasJS. 2004. Introduction to serum chemistries: artifacts in biochemical determinations. Inc Willard MD aind Tvedten H (eds.) Smail Animal Clinical Diagnosis by Laborstory Methods, 4" ed_ St Louis, Missouri: Elsevier. Tzeng SH, Ko WC, Ko FN, and Teng CM. 1991. Inhibition of platelet aggregation by some flavonoids. Thromb Res 64 {i 191-100. Accessed 5 April 2015. hitpsigoo.gl0G2ts Weiss DJ and Souza CD. 2010. Monocytes and macrophages and their disorders. In: Weiss DJ and Wardrop KJ (eds.). Schalm’s Veterinary Hematology. 6th ed. Ames, lowa: Wiley-Blackwell. *enaulis CG ang Steiner JM. 2010. Lipid metabolism and hyperlipidemia in dogs. Vet J Xenoulis PG. 2008. Investigations into idiopathic: hypertrigyceridemia in the Miniture Schnauzer in North America. Undergraduate Thesis. Veterinary Medicine and Biomedical Science, Texas A&M University. Hematology and biochemistry in dogs given malunagay a REFERENCES The preiiminary findings in this study suggest thet nanoliposome-encapsulated puslungusy enolic erat may fave a effet oo he platelets, Kidneys. and gid However, further studies should Ge done to verify ths association, Further studies on nanolpesome-encapsulated malunggay phenolic extract are recommended because of its potential in decreasing an elevated BUN level. However, caution should be taken in giving it a5 it may cause thrombocytopenia. Also, monitoring of the biochemical values is Tecommended since results may suggest the development of a liver problem. REFERENCES Aggamwal BB. 2010. Targeting inflammation-induced obesity snd metabolic diseases by curcumin and other nutraceuticals..Annu Rev Nuir 30: 172-199. Ajibade TO. Olzyem FO anc Arovioje ROA 2012, The heemetoiogical and biochemical effects of methanol extract of the seeds of Moringeoleifere in rats. J! Med Plants Fes (4} 615-521, Anderson KL. 2071. Litrasonography of Lipper Urinary Tract. Accessed 27 April 2018. ftp faoe.aiPASO) BarsantiJA, Lees GE, Willard MD and Green RA. 2004. Urinary Disorders. In: Willard MD and Tvedten H. Small Animal Clinical Diagnosis by Laboratory Methods. 4th ed. ‘St. Louis, MO: Elsevier. Behling-Kelly E. 2014. Serum lipoprotein changes in dogs with renal disease. J Vet Inter "ete oS: 1602-1803 Poe a "3 Choudhary M and Grower K. 2012. Development of functional food products in relation to obesity Func Foods Hith Dis 2(6): 188-197 Dua JS, Rana AC and Bhandari AK. 2012. Liposome: methods of preparation and applications. inf J Pharmaceut Stid Riss 3(2). 14-20. Ehrlich SD. 2011, Quercetin. Accessed 11 October 2013, htip:ligoo.gilvodkiy EmokpseAM snd Kuliys-GwareoA. 2014. Theinfuence of decreased levels of high density lipoprotein cholesterol on hematological indices in sickle cell disease patients. Ann Med Hith Soi Res 4 (2). 187-161. Fang Z and Bhandari 8. 2070. Encapsulation of polyphendls- a review. Trends Food Sci Tech 21: 510-623. German AJ. 2006. The growing problem of abesity in dags and cats. J! Muir 126 (Suppl): 19405-19455. 8 3B ¥ “3 (Supe Jain KS, Kathiravan MK, Somani RS and Shishoo CJ. 2007. The biology and chemistry of hyperlipidemia. Bioorg Med Chem 15: 4874-4899 Doi: 10.1016i_bme 2007 04031 Kalra i al definition and introduction. Am Assoc Pharmaceut Sci 5 Laflamme DP_ 19987. Development and validation of a body condition score system for dogs. Ganine Pract 22: 10-15. Mbikay M.2012. Therapeutic potential of Moringa oleifera leaves in chronic hyperglyoemia ov Td dyslinocenis: a tewew. Front Phana 9 Gay. 1-2, = McCullagh KG and Ehrhart LA. 1978, Non-srteriosoclerctic lesions in the kidneys ofdogs fed an atherogenic diet. Exp Ma Pathol 22: 400-418. Hematology and biochemistry in dogs given malunggay 43 start and atthe end of the study. The hematologic values and lipid levels of the dogs were compared from week 0, week 1, and week 5. Any changes from the initial kidney and liver function values were also compared for each dog RESULTS AND DISCUSSION Hematologic profile Table | shows the hematologic profile of dogs given given cholesterol with or without nanoliposome-eneapsulated or non-encapsulated malunggay phenolic esaract Table 1 Hematologic VaIU2E of dOgs given cholesterol whh or wimout nanclposome-encapsulsioc or non-sncapsulated malunggay phenolic extract administration. Control NonEM.o. Txt [ NLEM.o. Txt Normal Parameters ai | az | Bt Bz | ci | c range WO ms |i [32 [ies [ns | oss [sot Total WSC (e103 cella Wi wag | a4 | 01 [are [irs | ite | 5.0141 Wk 5 wa | 32 [733 [35 [a9 [i268 | sodas Wk o a7s [oss] woo [ was [ 797 | 612] 29420 Wk 7 me6 [sae] 717 | 202 | 42 | 719 [ 29120 Wks a61 [igi] sia | ges [ 73 | 743 | 29120 (cele 4 wea | 38 136 | a7 | 205 | zis | oa2s mphacytes 7 (cea weit | 236 |1.76| 172 | 442 | 28 | ies | 0229 wk S a3 3.64 253 77 369 3.65 042.9 Wik o o o o 204 o o 01-14 Neonocytee wet | o29 |o7e| 121 | 04 | 05a | 265] Otte (108 celieyuy wes | tao [o28[ 173 [ 0 oe | ust | 01-4 Wk o ooz | 15 | 026 | ots | oss | ove | 013 betas cea Wk T 0 a a 0 0 0 is Wks a a ozs | ot2| 0 O13 Wi G a ao | a o [ow] o eat fae Tae WT 0 0 a 0 0 0 bot Wks a a | a 0 ofa eat wee | 41 | a5 | 20 | 35 | 43 | 43 | 2537 POV (3) wet | 38 | 3 | 38 | 38 | #0 | 41 | 23557 wes | 20 | 40 | a7 | 39 | = | 28 | 557 WK G 15 [tss| 3 13 | 45 | os | s29 Pisielets (per 00x “we; [22 [aes] 15 | 255 | 45 | 195 | 629 oll Immereion tek) Wks 115 [17.5[ 65 2 7 #25 ae Garcia ef al. administration. Leukecytosis and neutrophilia were observed in dogs AZ and 82 Lymphocytosis was observed in all dogs except dog 81. Monocytosis was observed in dogs Al. 2. B1 and C2. Monocytopenia was observed in dog 82. These results suggest that the dogs may have experienced fear or excitement during bload collection (Schu 2010). However, stress and infection can also cause these abnormaities (Weiss and Souza, 2010). Thrombocytopenia was observed in dogs 81, C1, and C2 which suggests that dogs could haves neoplasia, immune-mediated thrombocytopenia, or pistee ines due tg Femonmage (Russel. 2010). However, tis may aise suggest tha the administration of nanoliposome-encapsulated malunggay phenolic exiract had an effect on the platelet count. This result is consistent with the study of Ajibade ef al. (2012) which shows that Sirisraton of meanal outatt of Al abies coeds hes jatelet lowering effect on rats. Kidney function tests Table 2 showed that the BUN level of dog C1 was elevated This suggests that the dog could have a gastrointestinal bleeding or fever. However, pre-renal azotemia is also.a possible cause of the elevation (Barsanti ef al, 2004}. itis important ta note, however, that the BUN levels ofdegs 0’ anc 02 were higher atthe satofthe stucy, Results alse showed that the serum creatinine levels of dogs 81 and B2 were elevated. This suggests that there is a decreased glomerular filtration. itis or muscle trauma is another ible cause but is very unikely (Barsanti ef al, 2004). Although elevated levels were observed. it was noted that the values on week 5 were lower compared to that of week 0, These results suggest that administration of malunggay phenolic extract containing quercetin, whether nanclipesome-encapsulated or not, may have an effect on the kidneys of dogs. This can be supported by the study of Shoskes (1008) wherein a similar effect was seen in the serum creatinine level of rats administered with quercetin. This is because of the ability of quercetin in preventing reperfusion injury by inhibiting peroxynttrites. Lipid profile Dogs At, A2, 61 and 82 had normal total cholesterol levels (125 to 300 madi} however, dogs C1 andC2both showed hypacholesteralemia( Table 3). rigiyeeridemia was alse observed in all dogs excapt dogs C1 and C2 which both had normal triglyceride Table 2, Blood urea nitrogen (SUN) and serum creatinine (CREA) values of dogs given cholesterol ‘wiih or without nandlipseome-eneapeulated or non-encapeulated malunggay phenolle extract BOMmintetragon. cavensters ‘Contra NonEM.o. Tet | NLEM.O. Txt | Normal Al az [ai [ae | ci [ ce | ore SUN mois weo [sea [ees [es [ear [es [eet [eee ‘ wes | 271 | 22a [asz | 202 | eee [ 52 | ees weo | 12 | 11 | 26 | 23 [ 13 [ 12 | 0547 SREA(ma'a wes [ta ii | 24 [22 | 12 i OST ‘Control filer capsules; NonEMLo. Txt-mon-2neapeulated malunggay phenalks extract, NLE M9. ‘Txt nanollposome-encapsulsies malunggsy pnenolle extract Philipp J Vet Anim Sci 2015, 41/1): 41-48 4 ORIGINAL ARTICLE HEMATOLOGIC PROFILE AND BIOCHEMICAL VALUES IN ADULT DOGS GIVEN CHOLESTEROL WITH OR WITHOUT NANOLIPOSOME-ENCAPSULATED MALUNGGAY (Moringa oleifera) ADMINISTRATION Cherissa A. Garcia’. Nelson B. Malabanan’, Marianne Leila 3. Flores". Benjamin Revel G. Marte! and Evelyn 6. Rodrigues? ABSTRACT ‘The hematologic profile and blochemical values In adult dogs given Nanollposoms-ancapelistsd malunggay (Moringa oleers) phanolle extract were atermingd In this study. Six adult dogs wera divided info tnree groupe: Group A: ‘cholesterol only; Group B: cholesterol with malunggay phenolic extract: and Grou ‘¢: chelseterol with nanolipoeome-encapaulsted metungasy phenolic extract, ‘two doge per group. Each dog wee subjected to 2 completa blood count at waeka 6. t and $ an serum biochemistry iaate st weeks 0 and 5. The results showed that ‘hrombocytopenia and g decresee In the elevated blood ures nitrogen leva were ob: administration of the nanoll gomeencapeulated maiungg Bhenollc extract. kalaa showed bslownormal chaleateral and lowdenelty ipapraten levels and within normal triglycerids and high denelty lipoprotein eve after the: experiment. Thess preliminary findings suggest that nanpliposome-2n apsulated malunggey phenallc extract may have a the platelets, kidnays, and lipid profile. However, further uae should be cone to vent this seecclation. *eywords: choleeterol, dog, hematology, kidneye, moringa, nanolipceom2 INTRODUCTION Dyslipidemia is the change in lipid levels (German, 2008). Dogs with chronic kidney disease cause a decrease in the high density li Ipopaten-choesirel (Behing Kelly 20 2014) which leads to higher levels of total WEC an and lower level rophils, lymphocytes. monocytes and packed cell volurne (Emakpae and Kullya-Gwarzo, 2014 Ayperlipidemis is the increase in the concentration of lipids in the blood more common associated with endocrine disorders, obesity and high fat diet that leads to pancreatitis, Secures and ocular and hepatic diseases (Xenoulis and Steiner, 2010). According to Jain ef al. (2007), 2 1% decrease in the total serum cholesterol decreases chronic heart disease

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