Sudan III Research Articles

THIN LAYER CHROMATOGRAPHIC TECHNIQUES FOR DETECTION OF SUDAN DYES IN PALM OIL

Sudan dyes I, II, III & IV are synthetic, oil-soluble, red-coloured azo dyes. These dyes are suspected genotoxic carcinogens and have been used in the adulteration of palm oil products. Many experimental studies on animal specimen have confirmed the formation of tumour due to the presence of different Sudan dyes in food products. In this study, a Simple Thin Layer Chromatographic method is described for the qualitative detection of Sudan-I, II, III & IV in palm oil gotten from different markets. The study includes extraction of sample, preparation of Thin Layer Chromatography (TLC) plates, Separation, detection and visualization of dyes on TLC plates. The dyes were extracted with solvent Acetonitrile. Sample extracts were decanted and analysed by simple thin layer chromatography in hexane, chloroform and acetic acid solvent system. The chromatograms of samples and standards were stained with Iron (ii) chloride stain and further evaluated in term of Rf values. Out of the five samples analysed the Rf values of the colouring spots from the three samples L2, L3 and L4, matched the standards (S2). It is therefore, concluded that samples L2, L3 and L4 contain Sudan IV dye. Without using any derivation, the limit of detection (18mg/L for S2 (Sudan IV dye) and 10mg/L for SI (Sudan III dye) was determined by visually determining if a spot was visible or not at a certain concentration of standard dye.

Glomerulosi tossica in febbre gialla

The present study is based on autopsic study of 70 fatal cases of yellow fever during the epidemic of 1952-53 in Costa Rica. Material was fixed in for­ maldehyde, Zenker's fluid, alcohol, and mercuric chloride solutions. The follow­ing stains were used in series: hematoxylin-eosin, Mallory, Van Gieson, Weigert (elastic fibers, fibrin) , Gallego, Feulgen, congo red, methyl violet, anilin blue, May-Gruenwald-Giemsa, Sudan III, and silver impregnation for reticulum. Observations revealed glomerular alterations produced in the course of yellow fever with hepato-renal syndrome, consisting of glomerulosic, Schiff­ positive transformation of the basal membrane or lamina densa, causing an altered permeability to proteins with infiltration (proteinuria) and establish­ment of an acute nefrosic syndrome. Such glomerulosic lesions are difficult to observe with hematoxylin-eosin alone, for which reason it has been held hitherto that the glomerulus is not affected in the hepato-renal syndrome.

Adult-onset generalized autoimmune enteropathy involving small intestine and colon: report of a case and review of literature

To investigate the clinicopathologic features of adult-onset autoimmune enteropathy (AIE). A case of adult-onset AIE was described along with a literature review. A 41-year-old male patient was admitted for intractable diarrhea for more than three months despite of any dietary restriction or anti-inflammatory therapy. Fat globule was observed by stool examination and Sudan III staining of the stool was positive. Enteroclysis showed weak movement and few plica of small intestine, while colonoscopy appeared normal. Small bowel biopsies revealed villus atrophy and increased crypt apoptotic bodies and lymphocytic infiltration in deep crypt. Although without significant surface intro-epithelial lymphocytosis, there were a large number of monocytes, lymphocytes, plasmacytes and neutrophilic granulocytes infiltrating in the lamina propria. Morphologically, the colonic mucous was similar to the small intestine although cryptitis and crypt abscess were significant in the former. Serum IgG anti-goblet cell antibody was demonstrated by indirect immunofluorescence. Other causes of diarrhea were excluded on the base of medical history, histopathology and other accessory examinations before the diagnosis of AIE was made. The patient had a complete remission after steroid treatment without recurrence for eight months during the follow-up even after steroid withdrawal for five months. AIE is exceedingly rare and timely diagnosis is important for successful therapy. Histological differential diagnoses should include ulcerative colitis, celiac disease, lymphocytic colitis, etc. The final diagnosis should be based on histological examination combined with the patient history, clinical manifestation, endoscopy finding and serological testing.

Effect of adenovirus shuttle vector-mediated RNA interference on steroid-induced adipogenic differ-entiation in the bone marrow mesenchymal stem cells of rabbits in vitro

Objective To investigate the effect of adenovirus shuttle vector-mediated RNA interferenee on steroid-induced adipogenic differentiation in the bone marrow mesenchymal stem cells (BMSCs) of rabbits.Methods BMSCs were obtained from bone marrow of New-Zealand rabbit,and divided into 6 groups at random:RNA interference groups 1,2 and 3 (S1,S2 and S3),irrelative sequence group (Con),model group (M) and normal control group (N).The BMSCs were treated with 1 × 10-7 mol/L dexamethasone and infected with small interfering RNA (siRNA) adenovirus shuttle vector.The expression of peroxisome proliferator activated receptor γ (PPARγ) mRNA was determined by TaqMan RT-PCR (TaqMan probe fluorescence detection),and expression of the PPARγprotein was determined by Western blotting.The content of intracellular triglyceride (TG) was determined.The cells were stained with Sudan Ⅲ,and adipocytes were counted under a light microscope.Results At 7th day after treatment,the expression levels of PPARγ mRNA in the BMSCs in S1,S2,S3,Con,M and N groups were 0.406 ±0.012,0.401 ±0.009,0.410 ±0.042,0.621 ±0.045,0.628 ±0.008 and 0.399 ±0.014 respectively,and those of PPARγ protein in blot grayscale scanning value were 0.246 ± 0.027,0.235 ± 0.015,0.257 ±0.025,0.800 ±0.017,0.793 ±0.019 and 0.244 ±0.010 respectively.The expression levels of PPARγ mRNA and protein in S1,S2,S3 and N groups were significantly lower (P ＜ 0.05) than those in M and Con groups.There was no significant difference (P ＞0.05) among S1,S2,S3 and N groups.The number of adipocytes and content of TG in S1,S2,S3 and N groups were significantly lower (P ＜ 0.05) than those in M and Con groups,but there was no significant difference (P ＞0.05) among S1,S2,S3 and N groups.Conclusion The adenovirus shuttle vector targeting PPARγ gene-mediated RNA interference can effectively inhibit steroid-induced adipogenic differentiation in the BMSCs of rabbits in vitro. Key words: RNA interference; Adenovirus shuttle vector; Steroid; Peroxisome proliferatoractivated receptor-γ; Adipogenic differentiation

Measurement of intrahepatic triglyceride stores by 1H magnetic resonance spectroscopy: study with rat models and in patients

To evaluate the accuracy of proton magnetic resonance spectroscopy (1H MRS) in quantitative measurement of intra-hepatic triglyceride (TG) stores in fatty liver at the levels of animal model and human body. 40 male Wistar rats were randomly divided into 2 equal groups: fatty liver model group, fed with high fat diet for 16 weeks, and control group, fed with standard diet. Every 4 weeks 5 rats from each group underwent 1H MRS to examine the intra-hepatic triglyceride contents and then were sacrificed. The liver tissues were stained with Sudan III] to detect the fat content. Eleven hospitalized patients with fatty liver suspected or confirmed by CT and with indications of liver puncture underwent 1H MRS and then liver puncture within 7 days. Linear regression was conducted to analyze the correlation between the results of 1H MRS and liver pathology. Intrahepatic lipid was detected in all the rat models, some rats of the control group, and the patients with fatty live. The lipid peak area and the ratio of the CH2 lipid peak area relative to the water peak area increased along with the duration of high fat diet feeding. In the rat models there was a close positive correlation between the hepatic TG content measured in vivo by 1H MRS and pathological measurement of liver fat content [r = 0.899; P <0001; slope 14.45; gamma-intercept -0.51]. The similar correlation existed in the patients too [r =0.814; P <0001; slope 33.72; gamma-intercept 27.04]. When the data of the rats and patients were analyzed together, the percentage of liver fat was highly positively correlated with the ratio of area under fat peak to the area under water peak (r = 0.878; P <0001; slope 56.33; gamma-intercept -0.68). A relative quantitative method to measure intrahepatic lipid content by 1H MRS, a safe, non-invasive method, is primarily established at the animal level and has been verified in humans with a good correlation with the pathological examination.

Fine needle aspiration cytology of eyelid sebaceous gland carcinoma and its differential diagnosis

To investigate the fine needle aspiration cytology (FNAC) features and differential diagnosis of eyelid sebaceous gland carcinoma. Four cases of eyelid sebaceous gland carcinoma diagnosed by FNAC were reported and confirmed by biopsy. Three of the cases were in early stages with tumor sizes smaller than 10 mm in diameter and without metastasis. The smears were stained by routine H & E and SudanIII methods. The cytologic findings were described and compared to corresponding histological features, and moreover, compared to chalazion, pilomatrixoma and eyelid basal cell carcinoma. Neither hemorrhage nor infection were found after the examination. Abundant cells were observed in the sebaceous carcinoma FNAC smears. Two types of tumor cells were found: one showed tumor cells differentiating toward sebaceous gland, with large pale cells and vacuolated cytoplasm, the other demonstrated poorly-differentiated cell with dark and irregular nuclei. Numerous vacuoles with inequality of size were found in cytoplasm or in background in all four cases, and the SudanIII stain showed that these vacuoles contained lipid. Some smears demonstrated cells with basaloid, fusiform or squamous features, corresponding to various histopathological types. In contrast, smears of chalazion displayed inflammatory granuloma, containing several types of inflammatory cells without malignant cells. Smears of pilomatrixoma were cellular with three cell populations, which included bland sheets of basaloid cells, nucleated basophilic cells and anucleated keratinized "ghost cells", along with calcific debris. The smears of basal cell carcinoma were typically less cellular, more tightly cohesive and had smaller clusters of uniform hyperchromatic basaloid cells without vacuolization in cytoplasm or background. Overall, the cytological features of eyelid sebaceous carcinoma were distinct from those of chalazion, pilomatricoma and basal cell carcinoma. FNAC is a safe and effective approach for the diagnosis of eyelid sebaceous carcinoma and lipid stain is useful in differential diagnosis. The application of FNAC may be important in reaching an early diagnosis and initial treatment of eyelid nodule.

P206 PPDとR-225の感作性と交叉反応性の検討

P206 PPDとR-225の感作性と交叉反応性の検討

Acute cytogenetic effect of benzene on rat bone marrow cells in vivo and the effect of inducers or inhibitors of drug-metabolizing enzymes

Acute cytogenetic effects of benzene in LE rat bone marrow cells in vivo were studied. Chromosome aberrations (CA) induced by benzene consisted mainly of gaps and breaks. Cells with exchanges were rarely observed. The incidence of benzene-induced CA was at its maximum level 12 h after the p.o. or i.p. administration of benzene, dependent on the dose of benzene administered, and higher in male rats than in female rats. However, the sex difference was not observed in the repeated inhalation experiment. Chromosome damage was higher with the p.o. than the i.p. administration. LE rats were more sensitive than Wistar and SD rats to the clastogenic action of benzene. Phenobarbital and Sudan III are well known as inducers of drug-metabolizing enzymes. The peak percentage of benzene-induced CA in the rats pretreated with phenobarbital was observed 6 h after the benzene injection, and it occurred at a higher level than in the rats given only benzene. On the other hand, Sudan III pretreatment suppressed benzene-induced CA at all periods after the benzene injection. SKF-525A (a cytochrome P-450 inhibitor) and cyclohexene oxide (an epoxide hydrase inhibitor) pretreatment also suppressed benzene-induced CA.

Effect of n-Alkanols on the Solubilization of Sudan III and Sudan IV in Aqueous Ionic and Nonionic Micelles

Solubilization of aminoazobenzene-beta-naphthol (Sudan III) and orthotoluene-azo-orthotoluene-azo-beta naphthol (Sudan IV) dyes in anionic, cationic, and nonionic micelles in aqueous medium, have been studied spectrophotometrically at 30°C. The absorbance of either of the solubilized dye in dilute surfactant solutions does not change appreciably. But it was found to vary linearly with surfactant concentration above the CMC. Comparable solubilities of dyes in concentrated solutions of surfactants and in organic solvents provide some indication about the location of the solubilizate in the micelle interior. For a particular dye-surfactant system, moles of surfactant solubilizing a mole of dye, N D , have been calculated from the straight live portions of the plots. N D has been found to depend on the nature of the surfactant and the structure of dye. Decreased values of N D with increased chain length of added alkanols have been explained in the light of partitioning of alcohol at the micelle water interface. Lower values of standard free energy of solubilization indicate smaller hydrophobic contribution in the solubilization process. The mechanism of solubilization has been found to depend on the nature of solubilizer, solubilizate, and the additives.

Studies with Sudan III in metabolism and inheritance

Studies with Sudan III in metabolism and inheritance