Anti-Cancer Effect of Moroccan Cobra Naja haje Venom and Its Fractions against Hepatocellular Carcinoma in 3D Cell Culture

Hepatocellular carcinoma (HCC) is the most common primary liver cancer in adults, the fifth most common malignancy worldwide and the third leading cause of cancer related death. An alternative to the surgical treatments and drugs, such as sorafenib, commonly used in medicine is necessary to overcome this public health problem. In this study, we determine the anticancer effect on HCC of Moroccan cobra Naja haje venom and its fraction obtained by gel filtration chromatography against Huh7.5 cancer cell line. Cells were grown together with WI38 human fibroblast cells, LX2 human hepatic stellate cell line, and human endothelial cells (HUVEC) in MCTS (multi-cellular tumor spheroids) models. The hepatotoxicity of venom and its fractions were also evaluated using the normal hepatocytes cell line (Fa2N-4 cells). Our results showed that an anti HCC activity of Moroccan cobra Naja haje venom and, more specifically, the F7 fraction of gel filtration chromatography exhibited the greatest anti-hepatocellular carcinoma effect by decreasing the size of MCTS. This effect is associated with a low toxicity against normal hepatocytes. These results strongly suggest that the F7 fraction of Moroccan cobra Naja haje venom obtained by gel filtration chromatography possesses the ability to inhibit cancer cells proliferation. More research is needed to identify the specific molecule(s) responsible for the anticancer effect and investigate their mechanism of action.

The Binding of Free and Sulfated Androstenone in the Plasma of the Boar

Androstenone circulates in the plasma bound to albumin before accumulating in the fat, resulting in the development of boar taint. Androstenone sulfate is more abundant in the circulation than free androstenone; however, it is unclear how androstenone sulfate is transported in the plasma and if steroid transport affects the development of boar taint. Therefore, the purpose of this study was to characterize the binding of androstenone sulfate in boar plasma and determine if variability in steroid binding affects the accumulation of androstenone in the fat. [3H]-androstenone sulfate was incubated with plasma and the steroid binding was quantified using gel filtration chromatography. Inter-animal variability was assessed by quantifying androstenone binding specificity in plasma obtained from boars that had high or low fat androstenone concentrations at slaughter. Androstenone sulfate bound minimally in the plasma and to isolated albumin, which suggests that it is transported primarily in solution. The specific binding of androstenone quantified in plasma and isolated albumin from low fat androstenone animals was significantly higher (p = 0.01) than in high fat androstenone boars. These results indicate that the binding of androstenone to albumin varies amongst individual animals and affects the transport of androstenone in the plasma and accumulation in the fat of the boar.

Screening for Macroprolactinemia Is Indispensable for Proper Treatment of Hyperprolactinemia Microadenoma

BACKGROUND: Macroprolactin microadenoma is a combination of non-functional microadenomas with macroprolactinemia. This study aimed to explore the significance of macroprolactinemia screening for the antidiastole of prolactin microadenoma and macroprolactin microadenoma.METHODS: Retrospective analysis of patients with pituitary microadenoma and screening for macroprolactinemia in patients with hyperprolactinemia was conducted. Based on the prolactin content and screening results, patients were divided into non-functional microadenoma, prolactin microadenoma, and macroprolactin microadenoma groups. The existing forms of prolactin in serum samples were analyzed by gel filtration chromatography and a luminescence immunoassay analyzer. The clinical course of patients and treatment were retrospectively reviewed.RESULTS: The results of gel filtration chromatography confirmed that prolactin in patients with macroprolactin microadenoma was mainly in the form of macromolecules, and the small molecular prolactin was within the normal range. Among 84 patients with hyperprolactin microadenomas, 9 (10.7 %) were diagnosed with macroprolactin microadenoma, and 5 (55.6 %) were treated with bromocriptine. The prolactin content (55.9 ~ 81.0 ng/mL) in the macroprolactin microadenoma group was elevated before the screening and significantly decreased (11.2 ~ 21.2 ng/mL) after screening. The incidence of clinical manifestations was the same as that of the non-functional microadenoma group, but the rate of drug use was higher, which was similar to that of the prolactin microadenoma group.CONCLUSION: Macroprolactinemia screening can effectively identify prolactin microadenomas and macroprolactin microadenomas, which prevent misdiagnosis and mistreatment of macroprolactin microadenomas.

ANALISIS HASIL FRAKSINASI PROTEASE DAN LIPASE YANG BERASAL DARI SALURAN PENCERNAAN UDANG VANAME (Litopenaeus vannamei)

Analysis of Protease and Lipase Fractionation Originated from the Digestive Tract of Vannamei Shrimp (Litopenaeus vannamei) Vannamei shrimp is a fishery commodity with a high consumption value, so it has an impact of high shrimp waste in the form of head and skin. The digestive tract connected to the head of the vaname shrimp (Litopenaeus vannamei) contains digestive enzymes, including proteases and lipases. This study aims to obtain the protein fraction that has the highest protease and lipase activity. The separation method used was centrifugation followed by precipitation using ammonium sulfate salt and dialysis. The dialysate was purified by gel filtration chromatography at a volume retention of 10 drops per tube. The proteolytic and lipolytic enzyme activity of the fraction was measured using a spectrophotometer. The results showed that fraction 102 had the highest protease activity value of 96.3924 U / mL, while fraction 100 had the highest lipase activity of 531.07 U / mL. This study showed that in the digestive tract of vaname shrimp, protease and lipase activity increased with the level of purity. Keywords: digestive enzymes, gel filtration chromatography, lipase, protease, vannamei shrimp ABSTRAK Udang vaname merupakan komoditi perikanan dengan nilai konsumsi yang tinggi, sehingga berdampak pula dengan tingginya limbah udang yang berupa kepala dan kulit. Saluran pencernaan yang terhubung dengan kepala udang vaname (Litopenaeus vannamei) mengandung enzim pencernaan, diantaranya protease dan lipase. Penelitian ini bertujuan untuk mendapatkan fraksi protein yang memiliki aktivitas protease dan lipase tertinggi. Metode pemisahan yang dilakukan adalah sentrifugasi dilanjutkan dengan pengendapan menggunakan garam ammonium sulfat dan dialisis. Dialisat dimurnikan dengan kromatografi filtrasi gel pada retensi volume sebanyak 10 tetes tiap tabung. Aktivitas enzim proteolitik dan lipolitik fraksi diukur menggunakan spektrofotometer. Hasil menunjukkan bahwa fraksi 102 memiliki nilai aktivitas protease tertinggi sebesar 96,3924 U mL–1, sedangkan fraksi 100 memiliki aktivitas lipase tertinggi sebesar 531,07 U mL–1. Penelitian ini menunjukkan bahwa pada saluran pencernaan udang vaname terdapat aktivitas protease dan lipase yang meningkat seiring dengan tingkat kemurniannya.

Biodegradation of Vegetable Oil Factory Effluent Using Extracellular Lipase Obtained from Alcaligenes Spp.

This study determined the degradation of vegetable oil factory effluent by extracellular lipase obtained from Alcaligenes spp. The extracellular lipase produced by isolated Alcaligenes spp was obtained and concentrated using Glycerol before Gel Filtration Chromatography. The partially purified enzyme obtained from the Gel Filtration Chromatography purification showed optimum activity at a temperature of 55° C and pH 7. The enzyme was then concentrated using glycerol prior to treatment. The raw effluent samples (from the vegetable oil factory) were treated with the partially purified enzyme for 20 days. The degradation activity of the enzymes on contaminant concentrations was monitored at every 5 days. A control experiment was also set up for possible natural degradation of contaminants. Effluents were analyzed for metals (Fe, Mn and Zn), Chemical Oxygen Demand (COD) and Lipid Content using the standard method. Data were analysed for graphical presentation using the Microsoft Excel package. Results showed reduction in Fe, Mn, Zn, COD and Lipid Contents between day 5 and 20 as 46.28%, 68.71%, 62.53%, 71.45% and 72.57% respectively. The application of extracellular enzyme in the treatment of the effluent enhanced the degradation of the effluent at a higher percentage than the natural attenuation process of the effluent. This showed the promising bioremediation potential of Alcaligenes spp.

Chromatographic Profiles of Umami Fractions from Indonesian Commercial Salty Soy Sauce

Salty soy sauce subjected in this study is a variety of commercial soy products in Indonesia. Chromatographic profiles linked to taste dilution analysis of the soy sauce were analyzed by Sephadex G-15 gel filtration chromatography followed by RP-HPLC. The results showed that there were 4 umami fractions (Fractions I − IV) obtained by Sephadex G-15 separation. Chromatographic profiles at 254 nm could show the differentiation of the four fractions and then their RP-HPLC profiles were proven to be different from each other. Fraction III which contained 65% of the soy sauce dry matters, had the highest umami intensity with umami TD factor of 256, meanwhile, this fraction was tasted salty due to the salt contained in the soy sauce. Fraction III was dominated by the later peaks in the RP-HPLC chromatogram, which was more hydrophobic. The hydrophobic components were commonly tasted bitter, perhaps in the commercial salty soy sauce, the taste interaction between the umami and bitter components might have occurred.

HIV-1 P24 EXPRESSION IN TRANSPLASTOMIC TOBACCO PLANTS

Expression of HIV-1 p24 gene in chloroplasts was achieved in a tobacco variety V2 (Virginia TBE2). Through PCR and Southern blot analyses, it was demonstrated that the transgene integrated into the target site in the chloroplasts, between trnfM and trnG. Western blot results showed that HIV-1 p24 gene expressed in transplastomic tobacco plants. p24 protein accumulations were detected by ELISA in the range from 1.7% to 6.3% TSP and the high concentrations in the leaves near the top. p24 protein was purified by gel filtration chromatography demonstrated that the purification is 9.694 folds and the performance is 31.94%, however, protein p24 largely was inactive after purification.

Discrepancy between Cardiac Troponin Assays Due to Endogenous Antibodies

Background Despite well-described analytical effects of autoantibodies against cardiac troponin (cTn) I on experimental assays, no study has systematically examined their impact on cTn assays in clinical use. We determined the effects of endogenous antibodies on 5 different cTnI assays and a cTnT assay. Methods cTn was measured by 6 methods: Siemens hs-cTnI Centaur, Siemens hs-cTnI Vista, Abbott hs-cTnI Architect, Beckman hs-cTnI Access, Beckman cTnI Access, and Roche hs-cTnT Elecsys. Measurements were repeated on 5 assays (all except Siemens hs-cTnI Vista) following immunoglobulin depletion by incubation with protein A. Low recovery of cTnI (<40%) following immunoglobulin depletion was considered positive for macro-cTnI. Protein A findings were validated by gel filtration chromatography and polyethylene glycol precipitation. Results In a sample of 223 specimens selected from a community laboratory that uses the Siemens hs-cTnI Centaur assay and from which cTn was requested, 76% of samples demonstrated increased cTnI (median, 88 ng/L; interquartile range, 62–204 ng/L). Macro-cTnI was observed in 123 (55%) of the 223 specimens. Comparisons of cTnI assays markedly improved once patients with macro-cTnI were removed. Passing-Bablok regression analysis between hs-cTnI assays demonstrated different slopes for patients with and without macro-cTnI. In patients with macro-cTnI, 89 (72%) showed no effect on the recovery of cTnT, whereas 34 (28%) had reduced recovery of cTnT. The proportion of results above the manufacturers' 99th percentile varied with the cTn assay and macro-cTnI status. Conclusion We suggest that the observed discrepancy between hs-cTnI assays may be attributed in part to the presence of macro-cTnI.