Characteristics of protein from red crab (Chionoecetes japonicus) shell by commercial proteases

This study was performed to examine the characteristics of protein of red crab (Chionoecetes japonicus) shell powder hydrolyzed by commercial proteases. Red crab shell was digested by commercial proteases, such as Protamex (P), Neutrase (N), Flavourzyme (F), Alcalase (A), Protease M (PM) and Protease A (PA). Protein yield analyzed by Biuret assay, absorbance at 280 nm and brix revealed that PA was the enzyme having the highest proteolytic activity. SDS PAGE showed that molecular weight of proteins produced by protease treatments was various and below 150 kDa. Combinational treatment of proteases (PA + P, PA + PM, PA + F, PA + A) was tried whether these increase protein hydrolysis from red crab shell powder compared to a PA single treatment. Soluble protein content was similar, but amino acid concentration by combinational treatments was higher than PA single treatment [PA + P 247.4 mg/g > PA + F (206.4 mg/g) > PA + A (133.4 mg/g) > PA + PM (59.1 mg/g) > PA (54.9 mg/g)]. Amino acid composition by combinational treatments was slightly different. Most abundant essential amino acids were phenylalanine, glycine, alanine, and leucine, whereas tyrosine and cystine were not detected.

Morphine Postconditioning Attenuates ICAM-1 Expression on Endothelial Cells

The purpose of this study is to determine 1) whether morphine postconditiong (MPostC) can attenuate the intercellular adhesion molecules-1 (ICAM-1) expression after reoxygenation injury and 2) the subtype(s) of the opioid receptors (ORs) that are involved with MPostC. Human umbilical vein endothelial cells (HUVECs) were subjected to 6 hr anoxia followed by 12 hr reoxygenation. Three morphine concentrations (0.3, 3, 30 microM) were used to evaluate the protective effect of MPostC. We also investigated blockading the OR subtypes' effects on MPostC by using three antagonists (a micro-OR antagonist naloxone, a kappa-OR antagonist nor-binaltorphimine, and a delta-OR antagonist naltrindole) and the inhibitor of protein kinase C (PKC) chelerythrine. As results, the ICAM-1 expression was significantly reduced in the MPostC (3, 30 microM) groups compared to the control group at 1, 6, 9, and 12 hours reoxygenation time. As a consequence, neutrophil adhesion was also decreased after MPostC. These effects were abolished by coadministering chelerythrine, nor-binaltorphimine or naltrindole, but not with naloxone. In conclusion, it is assumed that MPostC could attenuate the expression of ICAM-1 on endothelial cells during reoxygenation via the kappa and delta-OR (opioid receptor)-specific pathway, and this also involves a PKC-dependent pathway.

Development of a therapeutic method in the HPV-related cervical lesion using pH/temperature sensitive polymer spray formulation / 부인종양

OBJECTIVE: The causal link between oncogenic HPV(Human Papilloma Viruses) and the development of CIN (rvical intraepithelial neoplasia) and cervical cancer are now well established. Several medical therapeutic candidates aimd at the treatment of precancerous lesions and invasive carcinoma of the cervix. The objective of this study was to develop the pH-sensitive chitosan/alginate gels (pH=3.8-4.5) and temperature sensitive multiblock copolymers of PEG/PLA (poly (L-lactic acid)/polyethylene glycol) gels (temperature=37 degrees C) for controlled delivery of the paclitaxel (PTX). We had also evaluated whether PTX entrapped in chitosan/alginate gels or multiblock copolymers of PEG/PLA 1 could inhibit tumor growth in vivo. METHODS: PTX entrapped as microsphere in Chitosan/Alginate Microspheres were obtained using a spray-drying method. PTX-entrapped PEG/PLA gels were prepared by the solvent displacement method. We had prepared the multiblock copolymers of PEG/PLA which has the sol-gel-sol transition temperature at body temperature. The in-vivo efficacy of PTX in chitosan microphere or PTX in PEG/PLA mutiblock copolymer micelle were conducted in HeLa-tumor bearing Balb/c Nu/Nu athymic mice at an equivalent paclitaxel dose of 10 mg/kg with 48 hr interval. The inhibition of tumor growth was evaluated after 8 days of treatment. RESULTS: On 8 days after the transcutaneous treatment of PTX-containing chitosan microphere or PTX in PEG/PLA mutiblock copolymer micelle. significant inhibition in tumor growth was observed in balb/c nu/nu nude mouse carrying xenograft tumors (HeLa cells; HPV-18 positive state). Among these formulations, PTX in PEG/PLA mutiblock copolymer have shown improved therapeutic efficacy as compared to PTX-ivgroup. CONCLUSION: PTX-containing chitosan microphere or PTX in PEG/PLA mutiblock copolymer nanoparticles are a unique pH-sensitive and temperature sensitive drug delivery system. These formulations elicits enhanced efficacy as an effective and minimally invasive treatment in mice bearing human cervical cancer (HeLa Cells) xenograft.

Transcutaneous DNA immunization of HPV-16 E7 with hair-follicle specific promoter in mice / 대한산부인과학회지

OBJECTIVE: Previous reports have shown that transcutaneous immunization (TCI) with proteins or peptides in combination with adjuvants efficiently induces specific cellular and humoral immune responses. We compared the immune response after TCI with new construct which was derived from HPV-16 E7opt+K and pK6hf promoter instead of pCMV promoter and various adjuvant. METHODS: First, we made new construct ligated with HPV-16 E7 opt+K to Hair-follicle Specific pK6hf Promoter. Second, we applied pk6hf-E7 opt+K DNA with or without Lipofectamine 2000 and a combination of cholera toxin (CT) and CpG oligodeoxynucleotide (CpG) onto cold wax-depilated and hydrated bare skin of C57 BL/6 mice. To assess the ability of CTL(cytotoxic T-lymphocyte) activity, we performed intracellular cytokine staining with flow cytometric analysis to determine the number of E7-specific IFN-gamma- secreting CD8+ T cells generated in vaccinated mice with the DNA vaccine. RESULTS: Female C57BL/6 mice immunized by TCI methods with 30 microgram of pk6hf-E7 opt+K DNA with Lipofectamine2000 and CT efficiently generated E7-specific CD8(+) T cells compared with the group of pk6hf-E7 opt+K DNA only or DNA with Lipofectamine2000. CONCLUSION: Our results demonstrate that TCI of the linkaged-E7 DNA , E7 opt+K DNA to pk6hf, and Lipofectamine2000 and CT induced an antigen-specific CTL response. This result is of potential relevance for the development of therapeutic HPV-specific DNA vaccines with TCI and pK6hf promoter can be used safely.

CT Analysis of Lung Cancer and Coexistent Emphysema

PURPOSE: To evaluate the relation of the location and cell type of lung cancer to the location and degree in coexistent emphysema on high-resolution computed tomography (HRCT) scans. MATERIALS AND METHODS: Ninety-eight of 209 lung cancer patients having HRCT scans were retrospectively analyzed to assess the total lung emphysema and peritumoral regional emphysema. Single and primary lung cancers were included. The clinical data, including sex, age, smoking history and the pathologic cancer subtype, were recorded to correlate with the HRCT findings. The lobar distribution, central-peripheral predominance, surrounding parenchymal abnormality for cancer, cephalocaudal predominance, and subtype for emphysema were analyzed on HRCT. Using a CT scoring method, we scored the whole lung emphysema and peritumoral emphysema, and correlated the grading of emphysema with pulmonary functional values. RESULTS: Sixty-nine of 98 patients with lung cancer (71%) had emphysema. Lung cancer with emphysema was significantly higher in men than in women, and was significantly related to smoking. The mean age of cancer patients without emphysema was significantly lower than that of cancer patients with emphysema (68 yrs vs. 61 yrs, p=0.0006). Emphysema of grade I (0-25%) was found in 52 cases, grade II (25-50%) in 15, and grade III (50-75%) in 2. Total emphysema score was paralleled to peritumoral emphysema score in 64.3%, while the remaining patients had a higher peritumoral emphysema score (grade II or III) than total emphysema score (grade 0 or I). There was no statistical correlation in the developmental location between the emphysema and the lung cancer (significant correlation was only noted in grade II group of total emphysema score). The incidence of non-small cell carcinoma tended to be higher than that of small cell carcinoma in the two groups. CONCLUSION: The possibility of lung cancer in patients with pulmonary nodule, coexisting emphysema, and especially in elderly patients having a history of smoking must be clarified on HRCT. The location or type of lung cancer was not significantly correlated to the location or the degree of coexistent emphysema.

Cavernous Hemangioma of the Cavernous Sinus: A Case Report

Intracranial extra-axial cavernous hemangioma of the cavernous sinus is a very rare vascular malformation. It usually appears as a round non-encapsulated mass with well-defined borders, mimicking meningioma. We describe a case of cavernous hemangioma of the cavernous sinus, including the radiologic imaging findings, and also review the literature.