Cerebral ischemia is a serious neurological disorder that can lead to high morbidity and mortality. Baicalin is a naturally bioactive flavonoid derived from Scutellaria baicalensis Georgi, which has neuroprotective activity. Baicalin exerts a neuroprotective effect against hypoxic ischemic injury. In this study, we investigated whether baicalin regulates specific proteins in the cerebral cortex of ischemic stroke animals. Middle cerebral artery occlusion (MCAO) surgery was performed to induce ischemic brain injury, and baicalin (30 mg/kg) or vehicle was injected into the abdominal cavity before MCAO surgery. Neurological behavior tests were performed 24 h after MCAO surgery and proteomics approach was performed using proteins extracted from cortical tissue. Two-dimensional electrophoresis analysis and MALDI-TOF were performed to identify the regulated protein by baicalin. MCAO damage caused severe behavioral disorders, but baicalin treatment improved these behavioral deficits. Baicalin also induced changes in the expression of various proteins in the cerebral cortex of MCAO animals. Proteins changed by baicalin administration are as follow: adenosylhomocysteinase, isocitrate dehydrogenase [NAD] subunit alpha, apolipoprotein A-I, Rab GDP dissociation inhibitor beta, eukaryotic initiation factor 4A, and mu-crystallin. These proteins were involved in metabolism and protein synthesis. The results of this study demonstrated the neuroprotective effects of baicalin by improving behavioral disorders caused by MCAO damage. The results also showed that baicalin regulates the expression of a variety of proteins involved in neuroprotective functions. Therefore, our findings provide evidence that baicalin plays a neuroprotective role in stroke animal models by regulating specific proteins.
The primary therapeutic approach for Brucella species infections has mainly been based on antibiotic treatment. However, the development of vaccines for brucellosis control remains controversial. Furthermore, there is currently no licensed vaccine available for human brucellosis. This study aims to evaluate the effect of a combination of recombinant protein vaccines against Brucella (B.) abortus infection using a mouse model. Two B. abortus genes, namely dapB and gpm, were cloned and expressed in competent Escherichia (E.) coli DH5α using the pCold-TF vector. Successfully cloned vectors were subjected to PCR amplification using specific primer pairs. The apparent sizes of dapB and gpm were detected at 807 bp and 621 bp, respectively. Besides, the purified recombinant proteins dapB and gpm were detected using SDS-PAGE electrophoresis with correct sizes of 82.86 kDa and 87.61 kDa, respectively. These recombinant proteins were used to immunize mice as a combined subunit vaccine (CSV) to elicit host immunity against B. abortus infection. Mice immunized with CSV exhibited increased proliferation of CD4+ and/or CD8+ T cells at week 7th and 9th before sacrifice, in comparison to the control group. Notably, CSV immunization showed a significant decrease in bacterial burden in the spleen compared to the control group. Altogether, CSV using dapB and gpm induced host adaptive immune response against Brucella infection, suggesting its potential as an effective new subunit vaccine candidate.
This study was conducted to evaluate in vitro antioxidant activity of goat meat hot water extracts and the changes in apoptosis-related protein expression levels in the cancer cells treated with these extracts. Goat meat hot water extracts were prepared using different cuts of goat meat, including foreleg, hindleg, loin, and rib. Among these extracts, the foreleg and hindleg extracts displayed higher (P<0.05) ABTS radical scavenging activity than the other two extracts. Protein expression levels of BAX, p53, and p21 were not different in the cells treated with the extracts from different cuts, regardless of the cell type. Only p53 expression in HT-29 cells was elevated (P<0.05) after loin extract treatment. These results suggest that antioxidant activity and apoptosis-related effects of goat meat hot water extract varied with cut of meat under in vitro conditions. Because all data was obtained from the in vitro experiment, the ability to generalize conclusions is limited. Additional in vivo studies are necessary.
Ischemic stroke leads to severe brain damage and high mortality. Chlorogenic acid is a phenolic compound known to have neuroprotective properties. Bcl-2 family protein plays an important role in the regulation of apoptosis. We investigated whether chlorogenic acid exerts neuroprotective effects against ischemic injury by modulating Bcl-2 and Bax proteins. Middle cerebral artery occlusion (MCAO) was performed to induce cerebral ischemia and rats were injected intraperitoneally with phosphate buffered saline or chlorogenic acid (30 mg/kg) for 2 h after MCAO. Cortical tissues were collected 24 h after MCAO injury and reverse transcription-quantitative real time polymerase chain reaction and Western blot analyses were performed to investigate the expression of Bcl-2 and Bax. The regulation of Bcl-2 and Bax proteins by chlorogenic acid during glutamateinduced cell damage were examined. Cells were collected at 24 h after administration of glutamate (5 mM) and chlorogenic acid (10, 30, 50 μM). These results showed a decrease in Bcl-2 expression and an increase in Bax expression in MCAO animals, but chlorogenic acid treatment alleviated these changes by MCAO damage. Glutamate significantly reduced cell viability, and chlorogenic acid treatment alleviated this reduction in a dose-dependent manner. Glutamate induced a decrease in Bcl-2 expression and an increase in Bax expression, but chlorogenic acid treatment alleviated these changes. We found that chlorogenic acid alleviates changes in the expression of Bcl-2 and Bax proteins induced by brain injury. Therefore, our findings provide an evidence that chlorogenic acid has neuroprotective effects against MCAO damage by modulating Bcl-2 and Bax proteins.
Organisms constituting a large proportion of marine ecosystems, ranging from bacteria to fish, exhibit fluorescence and bioluminescence. A variety of marine organisms utilize these biochemically generated light sources for feeding, reproduction, communication, and defense. Since the discovery of green fluorescent protein and the luciferin-luciferase system more than a century ago, numerous studies have been conducted to characterize their function and regulatory mechanism. The unique properties of fluorescent and bioluminescent proteins offer great potential for their use in a broad range of applications. This short review briefly describes the functions and characteristics of fluorescent and bioluminescent proteins, in addition to summarizing the recent status of their applications.
Aluminum (Al) is one of the major factors adversely affects crop growth and productivity in acidic soils. In this study, the effect of Al on plants in soil was investigated by comparing the protein expression profiles of alfalfa roots exposed to Al stress treatment. Two-week-old alfalfa seedlings were exposed to Al stress treatment at pH 4.0. Total protein was extracted from alfalfa root tissue and analyzed by two-dimensional gel electrophoresis combined with MALDI-TOF/TOF mass spectrometry. A total of 45 proteins differentially expressed in Al stress-treated alfalfa root tissues were identified, of which 28 were up-regulated and 17 were down-regulated. Of the differentially expressed proteins, 7 representative proteins were further confirmed for transcript accumulation by RT-PCR analysis. The identified proteins were involved in several functional categories including disease/defense (24%), energy (22%), protein destination (9%), metabolism (7%), transcription (5%), secondary metabolism (4%), and ambiguous classification (29%). The identification of key candidate genes induced by Al in alfalfa roots will be useful to elucidate the molecular mechanisms of Al stress tolerance in alfalfa plants.
콩 [Glycine max (L.) Merr.]은 단백질, 지방, 탄수화물의 3대 영양소와 다양한 기능성 성분의 주요 공급원이다. 녹색종피와 자엽을 가진 콩 품종은 눈건강에 유익한 루테인 성분을 많이 함유하고 있지만, 항영양성분으로 알려진 쿠니츠트립신인히비터 단백질, 렉틴 단백질 및 P34 단백질을 가지고 있어 품질과 기능성을 저하시킨다. 이러한 성분이 유전적으로 제거된 품종의 육성이 필요하다. 녹색종피와 자엽을 가지고 있지만, 쿠니츠트립 신인히비터, 렉틴, P34의 3가지 단백질이 모두 없는 triple null 유전자형을 가진 콩 계통을 육성하기 위하여 본 연구가 진행되었다. 4개의 유전자원 (Gaechuck#2, PI548392, PI567476, Seonyack)을 이용하여 창성된 두 모본의 교배로 육종집단이 창성되었다. Western blot 기술을 이용하여 쿠니츠트립신인히비터, 렉틴 및 P34 단백질의 존재여부를 확인하였다. 녹색종피 및 자엽과 쿠니츠트립신인히비터, 렉틴, P34의 3가지 단백질이 모두 없는 triple null 유전자형 (titilelep34p34)을 가진 6개의 F2 종자가 선발되었다. F2 식물체 세대와 농업형질 평가를 통하여 한 개의 계통이 선발되었으며 F4 종자에서 3가지 단백질에 대한 유전적 고정을 확인하였다. 선발 계통의 경장은 73 cm 정도였으며 백립중은 19.5 g으로 중립에 속하였고 종피색은 녹색이고 제색은 검정색이며 성숙 자엽색은 녹색이었다. 본 연구를 통하여 선발된 계통은 성숙 콩 종실에서 품질과 기능성을 저하시키는 쿠니츠트립신인히비터, 렉틴, P34의 3가지 단백질이 모두 없으며 녹색종피와 자엽을 가진 유색콩 품종 육성을 위한 재료로 이용될 수 있을 것으로 사료되었다.
The objective of this study was to determine chemical compositions affecting the physical and thermal properties of the textured vegetable protein (TVP). The 14 commercial TVPs were pulverized, followed by analyzing their morphology, chemical composition, water absorption index (WAI) and water solubility index (WSI) (for the pulverized and original TVPs), solubility, swelling power, melting property, and hardness. All TVPs showed the rough surface with irregular cracks and pores and the porous structure with varied pore sizes. WAI was positively correlated to moisture and crude protein contents and negatively correlated to the total carbohydrate content. WSI and solubility were directly and reversely influenced by the crude ash and total carbohydrate contents and the crude protein and total starch contents, respectively. The swelling power and melting temperature of TVPs did not significantly affect chemical compositions. Melting enthalpies increased with crude ash content, while decreased with the total starch content. The hardness of the rehydrated TVPs was enhanced with their crude ash and total carbohydrate contents, whereas reduced with their crude protein and total starch contents. Overall, the yield and texture of the rehydrated TVP could be modulated with the crude protein and ash contents of TVP.