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Enzymatic preparation and antioxidant activities of protein hydrolysates from hemp (Cannabis sativa L.) seeds

  • Hyeon-Ji Yoon;Gyu-Hyeon Park;Yu-Rim Lee;Jeong-Min Lee;Hyun-Lim Ahn;Syng-Ook Lee
    • Food Science and Preservation
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    • v.30 no.3
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    • pp.434-445
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    • 2023
  • Hemp (Cannabis sativa L.) seeds have recently been attracting attention as a new high-value-added food material owing to their excellent nutritional properties, and research on the development of functional food materials using hemp seeds is actively progressing. This study aimed to evaluate the antioxidant properties of hemp seed protein hydrolysates. Protein hydrolysates were prepared from defatted hemp seed powder (HS) by enzymatic hydrolysis using five different proteases (alcalase, bromelain, flavourzyme, neutrase, and papain). 2,4,6-trinitrobenzene sulfonic acid (TNBS) assay and SDS-PAGE analysis revealed that HS showed a high degree of hydrolysis after treatment with each enzyme except papain. The total polyphenol content of the protein hydrolysates (<3 kDa) and the RC50 values obtained from two different antioxidant tests showed that alcalase hydrolysate (HSA) had a relatively high level of antioxidant capacity. In addition, treatment with HSA (25-100 ㎍/mL) significantly inhibited linoleic acid peroxidation. These results suggest that hemp seed protein hydrolysates are potential sources of natural antioxidants. Future studies will focus on the identification of active peptides from HSA.

Anti-Melanogenic Effect of Cannabis sativa Stem Extracts Fermented with Weissella paramesenteroides

  • Taehyun Kim;Jin-Woo Kim;Huitae Min;Jisu Park;Taejung Kim;Geun-Hyeong Kim;Byung-Joon Park;Jeong Kook Kim;Young-Tae Park;Jin-Chul Kim;Jungyeob Ham
    • Microbiology and Biotechnology Letters
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    • v.51 no.3
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    • pp.250-256
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    • 2023
  • Cannabis sativa (CS) has been in the spotlight not only for its medical uses but also as a raw material for cosmetics. As fermented cosmetics are known to have various health benefits, they have been extensively researched. Here, we investigated the characteristics of CS stems fermented using various gut microbes. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide assay and melanin content analysis revealed that melan-a cells containing CS stems fermented with Weissella paramesenteroides (CSWP) showed considerably reduced melanin content. Additionally, CSWP downregulated the expression of several melanogenesis factors, tyrosinase-related protein-1, and tyrosinase-related protein-2. This study suggests that the anti-melanogenic effect of CSWP could provide a new basis for the development of skin-lightening agents.

Predictors and Prevalence of Alcohol and Cannabis Co-use Among Filipino Adolescents: Evidence From a School-based Student Health Survey

  • Yusuff Adebayo Adebisi;Don Eliseo Lucero-Prisno III;Jerico B. Ogaya;Victor C. Canezo Jr.;Roland A. Niez;Florante E. Delos Santos;Melchor M. Magramo;Ann Rosanie Yap-Tan;Francis Ann R. Sy;Omar Kasimieh
    • Journal of Preventive Medicine and Public Health
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    • v.57 no.3
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    • pp.288-297
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    • 2024
  • Objectives: This study explored the prevalence and predictors of alcohol and cannabis co-use among 9263 Filipino adolescents, using data from the 2019 Global School-based Student Health Survey (GSHS). Methods: We conducted a cross-sectional secondary analysis of the GSHS, targeting adolescents aged 13-17 years and excluding cases with incomplete data on alcohol and cannabis use. Our analysis employed the bivariate chi-square test of independence and multivariable logistic regression using Stata version 18 to identify significant predictors of co-use, with a p-value threshold set at 0.05. Results: The weighted prevalence of co-users was 4.2% (95% confidence interval [CI], 3.4 to 5.3). Significant predictors included male sex (adjusted odds ratio [aOR], 4.50; 95% CI, 3.31 to 6.10; p<0.001) and being in a lower academic year, specifically grade 7 (aOR, 4.08; 95% CI, 2.39 to 6.99; p<0.001) and grade 8 (aOR, 2.20; 95% CI, 1.30 to 3.72; p=0.003). Poor sleep quality was also a significant predictor (aOR, 1.77; 95% CI, 1.29 to 2.44; p<0.001), as was a history of attempted suicide (aOR, 5.31; 95% CI, 4.00 to 7.06; p<0.001). Physical inactivity was associated with lower odds of co-use (aOR, 0.45; 95% CI, 0.33 to 0.62; p<0.001). Additionally, non-attendance of physical education classes (aOR, 1.48; 95% CI, 1.06 to 2.05; p=0.021), infrequent unapproved parental checks (aOR, 1.37; 95% CI, 1.04 to 1.80; p=0.024), and lower parental awareness of free-time activities (aOR, 0.63; 95% CI, 0.45 to 0.87; p=0.005) were associated with higher odds of co-use. Factors not significantly linked to co-use included age group, being in grade 9, always feeling lonely, having no close friends, being bullied outside school, and whether a parent or guardian understood the adolescent's worries. Conclusions: The findings highlight the critical need for comprehensive interventions in the Philippines, addressing not only physical inactivity and parental monitoring but also focusing on sex, academic grade, participation in physical education classes, sleep quality, and suicide attempt history, to effectively reduce alcohol and cannabis co-use among adolescents.

Histochemical Study of Secretory Lipids by Imidazole Staining (Imidazole 염색에 의한 분비지질의 조직화학적 연구)

  • Kim, Eun-Soo;Mahlberg, Paul G.
    • Applied Microscopy
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    • v.30 no.2
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    • pp.113-119
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    • 2000
  • Lipophilic glandular trichomes form secretory vesicles that accumulate in a distended noncellular secretory cavity . Imidazole-buffered osmium tetroxide solution was used to visualize unsaturated lipids in glands of Cannabis sativa. This method of staining revealed two kinds of secretory vesicles in the cavity of glands. Some smaller and rounded vesicles in the secretory cavity and secretory cells were positively stained with the imidazole, and they appeared electron-dense. The other type of vesicles which have bigger sizes more or less were not reacted, however, they appeared transparent. A high contrast of the cuticles which cover the gland was also strongly reacted with that processing. Those result suggest that the dark vesicles in the cavity may contribute to enlarge the subcuticular wall and cuticle when contents of these vesicles are dispersed into wall.

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Enzymatic saccharification of autohydrolyzed industrial hemp (Cannabis sativa L.) lignocellulosic biomass (자기가수분해 처리가 산업용 대마 목부 바이오매스의 효소 당화에 미치는 영향)

  • Shin, Soo-Jeong;Yu, Ju-Hyun;Lee, Soo-Min;Cho, Nam-Seok
    • 한국신재생에너지학회:학술대회논문집
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    • 2008.10a
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    • pp.74-76
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    • 2008
  • Autohydrolysis at different temperature levels was applied as industrial hemp pretreatment technique for glucose generation. Main structural components removed by autohydrolysis was xylan, which is more sensitive in acidic hydrolysis condition than cellulose or lignin. Higher temperature reaction conditions promoted more biomass components (xylan) removal than lower temperature, which led to better respond to enzymatic saccharification of residual biomass after autohydrolysis. With $185^{\circ}C$ and 60 min, saccharification degree was 53.0% of cellulose in hemp woody core biomass.

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Chemical characterization of the fast-growing industrial hemp (Cannabis sativa) woody core and bast fiber (산업용 대마의 목부와 인피섬유의 화학 조성 분석)

  • Shin, Soo-Jeong;Koo, Bon-Wook;Lee, Jae-Won;Choi, In-Gyu
    • 한국신재생에너지학회:학술대회논문집
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    • 2006.06a
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    • pp.482-484
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    • 2006
  • 대마에 대한 분석 결과, 인피 섬유의 경우 리그닌 함량은 7.6%로 다른 목질계 바이오매스나 일년생 초본류와 비교하여 매우 낮은 리그닌 함량을 나타냈으며 탄수화물 함량은 65.4%로 목질계 바이오매스와 유사하고, 초본류보다는 오히려 높았다. 목부는 리그닌 및 탄수화물 함량이 활엽수와 유사한 경향을 나타내었는데 특히 높은 자일란의 함량이 확인되었다. 또한 회분 함량이 인피섬유와 목부에서 각각 5.0%와 2.3%로 낮은 값을 나타내어 당화 및 발효 공정에 적용될 때 회분에 의한 공정 장애(스케일링 등)의 가능성을 낮추어 줄 것으로 사료되었다. 최종적으로 이러한 화학적 분석을 통해 대체에너지 생산을 위한 자원으로서 대마의 가능성을 확인할 수 있었다.

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Development and validation of a qualitative GC-MS method for THCCOOH in urine using injection-port derivatization

  • Sim, Yeong Eun;Kim, Ji Woo;Kim, Jin Young
    • Analytical Science and Technology
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    • v.34 no.2
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    • pp.68-77
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    • 2021
  • Cannabis is one of the most abused drugs in Korea. The main psychoactive component in cannabis, Δ9-tetrahydrocannabinol, is metabolized to 11-nor-9-carboxy-Δ9-tetrahydrocannabinol (THCCOOH) and THCCOOH-glucuronide (THCCOOH-glu) in the human liver, whereby the amount of THCCOOH-glu found in urine is twice as high as that of THCCOOH. The analytical process adapted by the majority of urine drug-testing programs involves a two-step method consisting of an initial immunoassay-based screening test followed by a confirmatory test if the screening test result is positive. In this study, a qualitative gas chromatography-mass spectrometry (GC-MS) method was developed and validated for the detection of THCCOOH in human urine, where THCCOOH-glu was converted into THCCOOH by alkaline hydrolysis. For purification of the urine extract prior to instrumental analysis, high-speed centrifugation was used to minimize interference. In addition, an injection-port derivatization method using ethyl acetate and N,O-bis(trimethylsilyl)-trifluoroacetamide containing 1 % trimethylchlorosilane was employed to reduce the time required for derivatization, and an aliquot of the final solution was injected into the GC-MS. The method was validated by measuring the selectivity, limit of detection (LOD), and repeatability. The sensitivity, specificity, precision, accuracy, Kappa, F-measure, false positive, and false negative rate were determined by comparing the GC-MS results with those obtained using the immunoassay. The LOD was determined to be 0.32 ng/mL, while the repeatability was within 9.1 % for THCCOOH. Furthermore, a comparison study was carried out, whereby the screening immunoassay exhibited a sensitivity of 86.4 % and a specificity of 100 % compared to GC-MS. The applicability of the developed method was examined by analyzing spiked urine and forensic urine samples obtained from suspected cannabis abusers (n = 221).

Verification of Biological Activities and Tyrosinase Inhibition of Ethanol Extracts from Hemp Seed (Cannabis sativa L.) Fermented with Lactic Acid Bacteria (대마씨 발효 추출물의 생리 활성 및 미백 활성 검증)

  • Yoon, Yeo-Cho;Kim, Byung-Hyuk;Kim, Jung-Kyu;Lee, Jun-Hyeong;Park, Ye-Eun;Kwon, Gi-Seok;Hwang, Hak Soo;Lee, Jung-Bok
    • Journal of Life Science
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    • v.28 no.6
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    • pp.688-696
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    • 2018
  • Hemp seed (Cannabis sativa L.; HS), an annual herbaceous plant in the Cannabis genus, has been reported to play various biological functions in immunity increase, atherosclerosis, constipation, hyperlipidemia prevention, anti-inflammatory, and anti-cancer. In recently years, as superfood, the growing interest in the health care benefits of hemp seed has led to increased consumption. In this study, we investigated the effect of an ethanol extract of HS fermented with lactic acid bacteria (Lactobacillus plantarum KCTC 3107, L. plantarum KCTC 3108, L. brevis BHN-LAB128, L. paracasei BHN-LAB129). An antibacterial activity against Staphylococcus aureus and Bacillus cereus were 13.99 mm and 15.17 mm, respectively. The ethanol extracts of fermented hemp seed by lactic acid bacteria that the contents of total polyphenol, total flavonoid content, DPPH radical scavenging activity, SOD-like activity, and ${\alpha}$-glucosidase inhibitory activity were increased compared to non-fermented hemp seed. Also, tyrosinase inhibitory activity of the fermented hemp seed (FHS), known to melanin increasing substance was increased. In these results, we suggested that FHS have effects of anti-oxidant, ${\alpha}$-glucosidase inhibitory activity, and tyrosinase inhibitory activity. Hence, we proposed that FHS has possible to development as functional foods and cosmetics.