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    • Javeid Iqbal et al.the fact that female urine normally contains 2-6 epithelial Krassner SM and Flory B (1972). Proline metabolism incells (intact or partially destroyed), (Brenner, 1983). These Leishmania donovani promastigotes. J. Protozol., 19:cells come from urogenetial tract and are considered 682-685.normally if not accompanied with pus cells or RBCs. In Lemesre JL, Darcy F, Kweider M, Carpon A and Santoro Furine these cells provide denatured proteins (mainly (1988). Requirement of defined cultivation conditions fornucleoprotein) which can enhance the growth of parasites. standard growth of Leishmania promastigotes in vitro. Acta. Tropica., 45: 99-108.REFERENCES Mukkada AJ (1977). Energy metabolism Leishmaniasis. Chang et al. (editors), Elsevier, Amsterdam, pp.49-65.Baily GG (1994). Visceral Leishmaniasis: More prevalent Mukkada AJ (1985). Energy metabolism in Leishmania in and more problematic. J. Infect., 29(3): 241-247 human parasitic disease, Leishmaniasis, Chang KP andBarral NM, Roters SB, Sherlock I and Reed SG (1987). Bray RS (editors). Elsevier Science Publishers, Destruction of Leishmania mexicana amazonensis, Amsterdam, New York and Oxford, pp.49-65. promastigotes by normal human serum. Am. J. Trop. O’Daly JA (1993). A comparison of molecular biology of Med. Hyg., 37(1): 53-V. Tryanosomes and Leishmaniae, and its impact on theBrenner BM and Reactor FC (eds) (1983). The Kidney, 2nd development of methods for the diagnosis and ed. Philadelphia, W.B. Saunders Co. vaccination of Leishmaniasis and Chagas diseases. Biol.Chang KP and Fish WR (1983).Leishmania: In vitro Res., 26(1-2): 219-224. cultivation of protozoan parasites. ed. Jensen JB. CRC Ray R and Ghosh AC (1980). Cultivation of Leishmania PRC Press, Coca Raton, Florida, pp.111-153. donovani in vitro in high yielding liquid culture medium.Donadio JV and Holley KE (1982). Membrane prolifrative Indian J. Med. Res., 71: 203. glomerulonephritis. Semin. Nephrol. 2: 204. Vouldoukis I, Alfred C, Monjour L, Mazier D, BrandicourtEvans DA (1986). An inexpensive easily available O, Ploton I, Tselentis Y, Nzuzi KK and Gentilini M replacement for foetal calf serum in media for the in vitro (1986). Cultures systems for productions of cultivation of Leishmania spp. Z. Parasitenkd., 72: 567- promastigotes and amastigotes forms of Leishmania. 572. Application to serological diagnosis and therapeuticHoward MK Pharoah MM, Ashall A and Miles MA (1991). trials. Ann. Parasitrol. Hum. Como., 61(2): 147-154. Human urine stimulates growth of Leishmania in vitro. Yasinzai MM, Iqbal J, Kakar JK, Ali SA, Ashraf S, Naz R, Trans Royal Soc. of Trop. Med. Hyg., 85: 447-479. Nasimullah M, Nagi AG and Salam A (1996).Kanwar YS (1884). Biopsychology of Glomrulus filtration Leishmaniasis in Pakistan: revised. J. Col. Phys. Surg. and protein urea. Lab Invest. 51: 7. Pak., 6: 70-75. Received: 26-1-2006 – Accepted: 3-4-2006REPORT PLASMA LIPID PROFILE IN SARCOMA PATIENTS M. IMRAN QADIR*, SALMAN AKBAR MALIK, ABDUL KHALIQ NAVEED* AND IJAZ AHMAD* Department of Biological Sciences, Quaid-i-Azam University, Islamabad *Department of Biochemistry & Molecular Biology, Army Medical College, Rawalpindi ABSTRACT Objective of the present study was to observe plasma lipid profile (triglycerides, cholesterol, LDL-cholesterol and HDL-cholesterol) in sarcoma patients. 120 subjects were included in the project. The subjects comprised of two groups; first as Controls (60 in number) and the second as Patients of Sarcoma (also 60 in number). Fasting blood samples were collected for estimation. Sarcoma patients showed highly significant (P<0.01) decrease, when compared with the normal control subjects. Keywords: Lipid profile, sarcoma.*Corresponding author: Email: mrimranqadir@hotmail.comPak. J. Pharm. Sci., 2006, Vol.19(2), 152-155 155
    • Plasma lipid profile in sarcoma patientsINTRODUCTION Procedure Three cuvettes were washed with distilled water and wereCancer of the connective tissues is known as Sarcoma. labelled blank, standard and sample. 20 µl distilled water,Sarcoma may be divided into different types according to its 20 µl standard and 20 µl sample, was pipetted in eachorigin (Robbins et al., 2003). cuvette respectively. Chromogen reagent, 2 ml was added toLipids are carried in body fluids with the help of each cuvette, contents of all the cuvettes were mixedlipoproteins (Edwards et, al., 1995 and Fischbach, 1984), thoroughly and incubated for 5 minutes at roomchylomicrons transport of triglycerides from intestine to all temperature. The wavelength of spectrophotometer was setcells. Very low density lipoproteins (VLDL) are involved in at 500 nm. Result command was given to spectrophotometerthe transportation of triglycerides from liver to other cells. and after some time results were displayed. The bloodLow density lipoproteins (LDL) are responsible for the triglycerides levels were calculated by applying thetransport of cholesterol from liver to the cells and high following formula.density lipoproteins (HDL) are involved for the transport ofcholesterol from cells to the liver. Chylomicrons and very Absorbance of samplelow density lipoproteins are rapidly catabolized (Heeren et, Triglycerides mg/dl = x 200al., 2003; Murray et al., 2000). Thus triglycerides, Absorbance of standardcholesterol, LDL–cholesterol and HDL–cholesterolconstitute Plasma Lipid Profile. Total Cholesterol Rapid enzymatic determination of the total cholesterol byResearchers have reported association of plasma/serum CHOD-PAP method, (Allian el al., 1974) was performed bylipids and lipoproteins with different cancers. As neoplastic using the commercially available kit manufactured bydisease is related to new growth, there is a greater utilization Human, Germany.of lipids including total cholesterol, lipoproteins andtriglycerides for new membrane biogenesis. Cells fulfill Procedurethese requirements either from circulation, by synthesis Three cuvettes were washed with distilled water and werethrough the metabolism or from degradation of major labelled blank, standard and sample. 20 µl distilled water,lipoprotein fractions like VLDL, LDL or HDL. The plasma 20 µl standard and 20 µl sample was pipetted in eachconcentrations of lipids are not the single additive function cuvette respectively. Chromogen reagent, 2 ml was added toof intake, utilization and biosynthesis because of its each cuvette. Contents of all the cuvettes were mixedcontinuous cycling in and out of the blood stream (Patel et thoroughly and incubated for 5 minutes at 37°C. Theal., 2004). wavelength of spectrophotometer was set at 500 nm. Result command was given to spectrophotometer and after someThe objective of the present study was to investigate any time results were displayed. The blood cholesterol levelsrelationship between plasma lipid profile (triglycerides, were calculated by applying the following formula.cholesterol, LDL-cholesterol and HDL-cholesterol) andsarcoma. Absorbance of sample Cholesterol mg/dl = —————————— x 200MATERIALS AND METHODS Absorbance of standardPatients LDL-CholesterolA total 120 individuals were included in our study. Out of LDL-cholesterol was determined by precipitation method.them 60 were normal subjects; 30 males and 30 females; Tests were performed by using the commercially availablehaving no cardiac or neoplastic disease. The remaining 60 kit manufactured by Randox, Germany.were patients of sarcomas. The patients had no other majorillness that affects plasma lipid profile. The patients were Procedurenot treated with any chemotherapy, radiation or surgery. For sample preparation; 100 µl sample and 1000 µlFasting blood samples were collected from CMH, precipitant were placed in a tube. After through mixing theRawalpindi. The plasma was stored at -20°C until used for tube was allowed to stand for 15 minutes at roomplasma lipid profile. temperature and then was centrifuged at 1500 rpm for 15 minutes. Supernatant was separated from the sediment andESTIMATION OF PLASMA LIPID PROFILE cholesterol was measured by the CHOD-PAP method. The LDL-cholesterol levels were calculated by applying theTriglycerides following formula.Triglycerides were determined by enzymatic method (GPO-PAP method), using the commercially available kit LDL-cholesterol mg/dl = Total cholesterol – Cholesterol inmanufactured by Human, Germany. supernatant.156 Pak. J. Pharm. Sci., 2006, Vol.19(2), 155-158
    • M. Imran Qadir et al. Table 1: Plasma lipid profile of control subjects and patients of sarcomas (Mean ± SD) Triglycerides Cholesterol LDL-cholesterol HDL-cholesterol (mg/dl) (mg/dl) (mg/dl) (mg/dl) Control subjects 149.87±14.03 171.47±19.52 73.30±10.17 50.27±9.26 Sarcoma patients 94.2±29.81 101.27±28.08 50.07±16.52 33.27±12.65HDL-Cholesterol 73.30±10.17 mg/dl and for HDL-cholesterol is 50.27±9.26HDL-cholesterol was determined by using the commercially mg/dl.available kit manufactured by Randox, Germany. In sarcoma patients, plasma level of triglycerides wasProcedure between 47-136 mg/dl with a mean value of 94.20±29.81.For sample preparation; 200 µl sample and 500 µl Plasma level of cholesterol was between 58-155 mg/dl withprecipitant were placed in a tube. After through mixing the a mean value of 101.27±28.08. Plasma level of LDL-tube was allowed to stand for 10 minutes at room cholesterol was between 28-71 mg/dl with a mean value oftemperature and then was centrifuged at 4000 rpm for 10 50.07±16.52. Plasma level of HDL-cholesterol was betweenminute. Supernatant was separated from the sediment and 18-66 mg/dl with a mean value of 33.27±12.65.cholesterol was measured by the CHOD-PAP method. Comparison between mean values of plasma lipid profile ofSTATISTICAL ANALYSIS control subjects and sarcoma patients is given in table 1. There is highly decrease in plasma levels of triglyceridesStatistical analyses were performed by using computer (37%) and cholesterol (41%); and moderate decrease inprogram SPSS 11.0 version. LDL-cholesterol (32%) and HDL-cholesterol (33%) in sarcoma patients. Thus all the plasma lipid componentsRESULTS AND DISCUSSION (triglycerides, cholesterol, LDL-cholesterol and HDL- cholesterol) of sarcoma patients showed highly significantIn the present study, plasma level of triglycerides in control (P<0.01) decrease, when compared with the normal controlmales was between 132-178 mg/dl with a mean value of subjects. Robertson and Ray, (1919) decided that frequency149.67±13.57. Plasma level of cholesterol in control males of the incidence of sarcoma is reduced by the administrationwas between 135-208 mg/dl with a mean value of of cholesterol.171.40±19.64. Plasma level of LDL-cholesterol in controlmales was between 54-57 mg/dl with a mean value of Lipids are major cell membrane components essential for73.47±8.82. Plasma level of HDL-cholesterol in control various biological functions including cell growth andmales was between 35-62 mg/dl with a mean value of division of normal and malignant tissues. Low levels of48.00±7.54. cholesterol in the proliferating tissues and in blood compartments could be due to the process of carcinogenesis Plasma level of triglycerides in control females was (Patel et al., 2004).between 129-179 mg/dl with a mean value of 150.07±14.01.Plasma level of Cholesterol in control females was between CONCLUSION138-201 mg/dl with a mean value of 171.33±18.77. Plasmalevel of LDL-cholesterol in control females was between This study has shown that plasma lipid levels are decreased52-95 mg/dl with a mean value of 73.13±11.05. Plasma in sarcoma patients. As there is a change in plasma lipidlevel of HDL-cholesterol in control females was between profile of Sarcoma patients, the plasma lipid profile may be42-70 mg/dl with a mean value of 52.20±9.73. helpful for diagnosis of the disease.Comparison between mean values of plasma lipid REFERENCESprofile of control males and control females showedstatistically no significant (P>0.05) difference. Thus Allian CC, Poon LS, Chan CS and Richmond W (1974).mean of the two were taken as reference values. The CHOD-PAP method for determination of totalreference value for triglycerides is 149.87±14.03 mg/dl, for cholesterol, Clin. Chem., 20: 470.cholesterol is 171.47±19.52 mg/dl, for LDL-cholesterol isPak. J. Pharm. Sci., 2006, Vol.19(2), 155-158 157
    • Plasma lipid profile in sarcoma patientsEdwards CRW, Baired JD, Frier BM, Shephered J and Toft Patel PS, Shah MH, Jha FP, Raval GN, Rawal RM, Patel AD (1995). Ischaemic heart disease. In: Davidsons MM, Patel JB and Patel DD (2004). Alterations in plasma Principles and Practice of Medicine, edited by Edwards lipid profile patterns in head and neck cancer and oral CRW, Boucher JAD, Haslett C and Chilvers E, 17th ed., precancerous conditions. Indian J. Canc., 41(1): 25-31. ELBS, Churchill Livingstone, London, pp. 245-66. Robbins SL, Cotran RS and Kumar V (2003). Neoplasia, In:Fischbach FT (1984). Chemistry Studies. In: A Manual of Robbins Basic Pathology, 7th ed., Saunders, Philadelphia, Laboratory Diagnostic Tests, 2nd ed., JB Lippincott USA, pp.165-210. Company, Philadelphia, pp.223-358. Robertson TB and Ray LA (1919). Lesions exhibited byHeeren J, Grewal T, Laatsch A, Rottke D and Rinninger F normal, pituitary -, lecithin -, cholesterol - and tetheli - (2003). Recycling of apoprotein E is associated with fed white mice at the occurrence of natural death, with cholesterol efflux and HDL internalization, J. Bio. Chem., especial reference to the incidence and development of 278(16): 14370-78. spontaneous cancer. In: Experimental studies on growth,Murray RK, Granner DK, Mayes PA and Rodwell VW www.jbc.org., pp.443-53. (2000). Lipid transport and storage, Appendix. In: Harper’s Biochemistry, 25th ed., Appleton & Lange, Received: 28-12-2005 – Accepted: 10-04-2006 USA, 268-84 and 867-72. REVIEW POROUS NANOPARTICLES IN DRUG DELIVERY SYSTEMS M. SAEED ARAYNE AND NAJMA SULTANA* Department of Chemistry, University of Karachi, Karachi-75270, Pakistan *Research Institute of Pharmaceutical Sciences, Faculty of Pharmacy, University of Karachi, Karachi-75270, Pakistan ABSTRACT This article concentrates mainly on fabrication of porous nanoparticles, its characterisation and its use for controlled release of drug. It also encompasses the strategies that have been used to translate and fabricate a wide range of particulate carriers e.g., nanospheres, liposomes, micelles, oil-in-water emulsions, with prolonged circulation and/or target specificity. Sol-gel technique is one of the most widely used techniques to fabricate porous nanoparticles within the polymer. Such nanoparticles have also applications in vascular drug delivery and release, site-specific targeting, as well as transfusion medicine. With regard to the targeting issues, attention is particularly focused on the importance of physiological barriers. We have also critically reviewed and assessed the fate and activity of biodegradable polymeric drug delivery vehicles because the uniformity in degradation of these polymers is questionable. This article will highlight rational approaches in design and surface engineering of nanoscale vehicles and entities for site-specific drug delivery. Potential pitfalls or side effects associated with nanoparticles are also discussed. Keywords: Nanotechnology; nanoparticles; nanofibers; controlled-release; nanofabrication; biopharmaceuticals; porous nanoparticles; nanosized drug delivery systems; macrophage; endothelium; intracellular delivery; extravasation; toxicity; antituberculosis drugs; nanoparticles tuberculosis therapy. intracellular internalization; endocytosis; bone marrow differentiation. *Corresponding author: Tel.: +92-21-4610132; email arayne@gawab.com158 Pak. J. Pharm. Sci., 2006, Vol.19(2), 155-158