Iranian Journal of Basic Medical Sciences
Year:2003 | Volume:6 | Issue:1 (17)|Papers Count:9

The aim of this study was to evaluate the diagnostic value of AgNOR (Argyrophilic Nucleolar Organizer Proteins Region) staining in differentiation between malignant and non-malignant lesion, and histological grading of bladder transitional cell carcinoma. We have also showed the rate of AgNOR distribution score in the nuclear of malignant and non-neoplastic cells of bladder epithelium as an important diagnostic marker at light microscopic level.Forty nine paraffin block samples selected from the Pathology Department of Ghaem Hospital including 6 chronic cystitis, 5 transitional cell carcinoma (Grade I), 15 transitional cell carcinoma (Grade II) and 23 transitional cell carcinoma (Grade III). Nucleolar AgNOR dots were studied in epithelial cells of chronic cystitis and transitional cell carcinoma of bladder Grades I, II, III by a light microscope to distinguish the average number of AgNOR dots and their distribution rate in nuclei of 100 cells which had three or more AgNOR dots. There was a significant stepwise increase in the percentage of cells exhibiting nucleoli with at least three or more distinct silver dots in chronic cystitis and transitional cell carcinoma of bladder Grades I, II, III. The counting of AgNOR dots and determining their distribution score are simple, suitable and cheap method to distinguish malignant neoplastic and non-neoplastic lesion of bladder. This method has also a high diagnostic value for grading of bladder transitional cell carcinoma.

In this in vivo study, 120 adult patients were selected by BMI factor (30 patrents in each method) for measuring thyroid dose. CT scan methods included conventional CT and spiral CT with three different pitch factors (1.2, 1.5, 1.7). By use of TL dosimetry, thyroid dose due to scattered radiation in thorax CT scan was measured and the mean of values in each method compared by t-test with reliability of 95% (α=0.05). Dose values also normalized by BMI length of scan (L), patient weight (W). BMI and length of scan together and compared by t-test with reliability of 95% (α =0.05). Results indicates spiral CT with pitch factor greater than one, reduces thyroid dose in comparison of conventional CT and also increase of pitch factor decreases thyroid dose.

Alginate is a safe, non-immunogenic and inexpensive natural polymer with high mucoadhesive properties. Alginate microspheres can be used as a delivery system for antigens to the mucosal surfaces. Alginate microspheres were prepared by emulsification-internal phase gelation. Effects of different sonication times; alginate, emulsifier and calcium concentrations; and the volume of calcium solution on mean size, size range, surface roughness and porosity, sphericity and clumping of microspheres were evaluated using optical microscope and zetasizer. The best conditionswere resulted by 90 sec. sonication, 3% alginate solution, 2% surfactant, and 60 ml of 0.33% CaCl2 in octanol. The resulting microspheres had mean size of 1.34±0.3 μm (n=3) and size range of 0.3 - 2.0 μm with no surface roughness and porosity, low clumping and high sphericity. The encapsulation rate was about 47.7% (n=6). All batches showed nearly the same release profiles with a low burst release. The intacticity of model antigen (tetanus toxoid=TT) extracted from microspheres was confirmed by SDS-PAGE and the antigenicity of TT was studied by ELISA and found to be 91±5% that of the original TT. With regard to the size and morphological characteristics of the prepared microspheres and preservation of the antigenicity of the encapsulated TT, they could be used as a delivery system for mucosal delivery of TT.

The main objective of this study was to formulate a liposome preparation loaded with recombinant major surface metalloproteinase glycoprotein of Leishmania (rGP63) to selectively induce cell mediated immunity (CMI) in susceptible BALB/c mice and protect the mice against challenge with virulent L. Major. rGP63 was incorporated into liposomes composed of lecithin and cholesterol, 1:1 molar ratio by solvent evaporation (Multilamellar vesicles, MLV-rGP63), reverse phase evaporation (Large unilamellar vesicles, LUV-rGP63) and detergent solubilization (Small unilamellar vesicles, SUV-rGP63) procedures. Liposome preparations containing rGP63 (2 µg), rGP63 (2 µg) by itself, PBS (Phosphate buffer saline), and a control liposome (rGP63-free liposomes) were injected subcutaneously (SC) three times in groups of 10 female BALB/c mice with three week intervals. Three weeks later the mice were tested for delayed type hypersensitivity (DTH) by injecting 2 µg of rGP63 SC to the left footpads; and for comparison the right footpads were injected with PBS. After 24, 48 and 72 hours the footpad thickness was measured on both foot pads. One week after the DTH test, the mice were challenged with virulent L.major promastigotes (1×106 / 50 µL) SC to the left footpads and the footpad thickness was measured for 10 weeks. The results of DTH test indicated that among different groups, the SUV-rGP63 had the greatest positive DTH response compared to the control groups (p<0.01). The DTH response of LUV-rGP63 and rGP63 groups were also positive (p<0.05); however, the response of MLV-rGP63 was less than the other groups and it was significant (p<0.05) only at 24 and 48 hours compared to control groups. In the challenge test, the SUV-rGP63 liposomes had a complete protective effect (p<0.001), the LUV-rGP63 and MLV-rGP63 had a little protective effect that was not statistically significant, and rGP63 by itself showed only partial protection. These results indicate that liposomes prepared by detergent solubilization procedure (SUV-rGP63) could be a suitable immunoadjuvant for rGP63 to induce selective CMI in susceptibleBALB/c mice and protect the mice against cutaneous leishmaniasis.

Esophageal cancer is very common in some geographical regions of Iran. This area is referred to as the belt of esophageal cancer that extends from northern Iran to north of China. Human papilloma virus has been suggested as an etiologic factor for esophageal cancer. Many studies in the world, especially in regions with high prevalence, have shown the co-existence of this virus with esophageal squamous cell carcinoma (SCC). Paraffin-embedded archival tissues were collected for this research project, from 1997 to 2002, to evaluate the presence of HPV genome in samples from esophageal SCC. Simultaneously, the p53 mutant protein expression was assessed to find the relation between HPV infection and p53 mutant protein expression. For this study, 50 cases of esophageal SCC paraffin blocks from Ghaem University Hospital in Mashhad were chosen from 1997-2002. After DNA extraction, various genetic types of HPV genome (types 16 and 18) were detected using specific primers in a PCR assay. 8/45 testable cases (20 %) were positive for HPV type 16 and all the samples were negative for HPV type 18. P53 mutant protein was detected by Immunohistochemistry in 37/50 cases (74 %). Interestingly, all HPV positive samples were negative for p53 expression that was compatible with the reported HPV cancerogenesis. HPV E6 protein binds to and causes degradation of p53 protein exempting the deregulated cell to undergo p53-mediated control. Our results indicate that HPV may be a factor in pathogenesis of esophageal SCC. This project was supported by the research council of Mashhad University of Medical Sciences.

Ginger is the root part of Zingiber of Jicinale that contains numerous ingredients including oleoresins, zingiberen, graniol, gingerol, pyrogalols, and hydroxyalkanone. It has been reported that hydroxyalkanone is responsible for the antinausea and antivomiting effect of ginger. In this study, the embryotoxicity effect of aqueous and ethanolic extracts of ginger was evaluated, applying the standard GLP protocol for this purpose. Maximum tolerated dose of the two extracts were determined and appropriate doses were administered intraperitoneumaly in time-pregnant mice on days six to fourteen of gestation. Embryos were harvested by caesarian section on day 18, examined externally and then stained by a microwave technique. Both extracts caused significant retardation in the weight gain of the pregnant mice and a notable reduction in uterus weight. Weight and crown-lump length of treated embryos were reduced. Both extracts caused many skeletal and tissue malformations, which clearly indicated the teratogenic potential of ginger extracts at administered doses. It is suggested that ginger's reported effects on smooth muscle, leukotriens and prostaglandins synthesis could be a major factor in producing these toxicities. This plant is used extensively in traditional medicine and based on this study, its use during pregnancy should be restricted until further investigations.

Extracellular matrix and cell-coat complex carbohydrates play a major role in critical cellular functions. The purpose of the present study was to investigate the role of certain glycoconjugates during transition of the limb bud core mesenchymal cells into cartilage. Using histochemicalprocedures, embryonic mouse limb buds from days 10 to 16 of development were processed and examined with HRP-lectins including DBA, VVA, VVA/B4 and SBA, to localize special glycoconjugates containing terminal N-acetylgalactosamine. The results showed striking reaction of only VVA and VVA/B4 with the core limb condensed mesenchyme. The timing and distribution of staining with VVA and VVA/B4 suggest that specific terminal sugar may play a role in transition of mesenchymeinto core condense cells and finally into pre-cartilage stage and subsequent formation of the cartilage.

The current work provides information on the surfactant induced membrane permeability. The membrane model was a Franz diffusion cell with shed snake-skin as membrane. Also, liposomes made by cholesterol and phosphatidylcholine, prepared by freeze-thawing method were tested as synthetic lipid membrane models. Three hydrophilic molecules, gentamicin sulfate, 5(6)-carboxyfluoresceine and phenol red were used as drug models. Absorption enhancers were sodium lauryl sulfate (SLS), benzalkonium choride (BC), that are anionic and cationic surfactants, respectively, and Quillaja saponaria (QTS) and Acanthophyllum squarrusom (ATS) total saponins, two naturally origmated enhancers. Surface tension changes and hemolytic activities were also determined to find out any probable relationship between surfactant properties and its enhancingactivity. The obtained results showed that maximum enhancing effect occured below the surfactant critical micelle concentration (cmc), while they reach their maximum hemolytic activity with concentrations more than their cmc. The results also indicated that there was no enhancing effect induced by benzalkonium choride when applied on the skin model, probably due to the similarity of electric charges on the skin surface and surfactant ions. Among surfactants tested, SLS showed maximum enhancing effect in the diffusion cell model, while benzalkonium choride had maximum activity in liposome membrane model. Both QTS and ATS showed a moderate activity in both diffusion cell and liposome models. The precise enhancement mechanism of surfactants tested in this study is not clearly known.

The sciatic nerve of 24 adult male wistar rats were exposed and transected in the mid thigh (gluteal muscles region). Rats were divided in 2 experimental and control groups. The effect of single dose of Fetal Brain Extract (FBE) treatment on motoneuron survival was investigated experimentally and compared with untreated (control group). This study showedthat by 6 weeks post-injury, nearly 80% of motoneurons survived in experimental models (treated by FBE), while only about 40% of motoneurons survived in control animals (nerve transected only). This data indicates that treatment with a single dose of FBE at time of ingury can promote the lesioned motoneurons in anterior horn of spinal cord of experimental significant in comparison with control group.