Bucillamine prevents cisplatin-induced ototoxicity through induction of glutathione and antioxidant genes.

Bucillamine is used for the treatment of rheumatoid arthritis. This study investigated the protective effects of bucillamine against cisplatin-induced damage in auditory cells, the organ of Corti from postnatal rats (P2) and adult Balb/C mice. Cisplatin increases the catalytic activity of caspase-3 and caspase-8 proteases and the production of free radicals, which were significantly suppressed by pretreatment with bucillamine. Bucillamine induces the intranuclear translocation of Nrf2 and thereby increases the expression of γ-glutamylcysteine synthetase (γ-GCS) and glutathione synthetase (GSS), which further induces intracellular antioxidant glutathione (GSH), heme oxygenase 1 (HO-1) and superoxide dismutase 2 (SOD2). However, knockdown studies of HO-1 and SOD2 suggest that the protective effect of bucillamine against cisplatin is independent of the enzymatic activity of HO-1 and SOD. Furthermore, pretreatment with bucillamine protects sensory hair cells on organ of Corti explants from cisplatin-induced cytotoxicity concomitantly with inhibition of caspase-3 activation. The auditory-brainstem-evoked response of cisplatin-injected mice shows marked increases in hearing threshold shifts, which was markedly suppressed by pretreatment with bucillamine in vivo. Taken together, bucillamine protects sensory hair cells from cisplatin through a scavenging effect on itself, as well as the induction of intracellular GSH

Biochemical properties of a novel neoagarotriose-producing β-agarase from Gilvimarinus agarolyticus JEA5

An agar degrading bacterium was isolated from seawater, collected from the east coast of Jeju Island, republic of Korea and identified as Gilvimarinus agarolyticus JEA5. The β-agarase gene from Gilvimarinus agarolyticus JEA5 (rGaa16B) was identified from draft genome sequence by BLAST. Gaa16B has 1800 bp of open reading frame encoding 636 amino acids (aa), and include glycosyl hydrolase family 16 (GH16) β-agarase module and two carbohydrate binding module 6 (CBM6). The Gaa16b was cloned and overexpressed as a MBP-fusion recombinant β-agarase (without signal peptide and two CBM6) in E. coil. rGaa16B showed highest activity at 60°C and pH 7. After incubation at 45OC for 90 min, rGaa16B showed over than 95% of its initial activity. rGaa16B were enhanced in the presence of MnCl2, KCl2, MgCl2, FeSO4. rGaa16B showed 2112.1 unit/mg in the presence of 2.5 mM of MnCl2. rGaa16B produce mainly neoagartetraose (NA4) and neoagarobiose (NA2). Interestingly, we observed neoagartriose (NA3) from hydrolytic products of rGaa16B. LC/Mass analysis was performed to confirm the hydrolytic products containing neoagarotriose. We found three different hydrolytic products which showed 324.28, 468.41, 630.55 Da of molecular weight, respectively. Please click Additional Files below to see the full abstract

Recombinant protein production in Escherichia coli by combining of signal peptide originated from Bacillus subtilis

We isolated chitosanase secreting B. subtilis CH2 and identified the chitosanase nucleotide sequence. Analyzed the sequence showed that it consisted of 813 bp, including 87 bp signal sequence. The signal sequence leads the target protein to the cell-membrane of the B. subtilis CH2 and then secret the chitosanase out of the cell. The signal peptide showed 6 amino acids deletion compared to other B. subtilis chitosanase signal peptides. The chitosanase sequence including signal peptide was cloned into pET11a vector without fusion and expressed in E. coli BL21(DE3). The expressed chitosanase in E. coli showed two distinct bands which represent the pro-chitosanase in cytoplasm and mature chitosanase in periplasm. Time frame induction and results showed that muture chitosanase was increased. Subsequently, we linked this chitosanase signal sequence in front of B. subtilis CH2 xylanase and human superoxide distimutase 1 (hSOD1) sequences, and expressed it in E. coli BL21(DE3). The recombinant xylanase and hSOD1 moved to periplasmic space with high efficiency. This signal sequence is useful for bio-medical protein production in E. coli. Please click Additional Files below to see the full abstract

Prenatal screening for neural tube defects: from maternal serum alpha-fetoprotein to ultrasonography

The two main screening tests during pregnancy are those for chromosomal abnormalities and neural tube defects (NTDs). In particular, for NTDs, measurement of maternal serum alpha-fetoprotein (MSAFP) levels early in the second trimester (15–18 weeks of gestation) has been considered the gold standard screening test for the past 4 decades. However, with remarkable technological advancements and the widespread use of ultrasound during those periods, mid-trimester ultrasonography has gradually replaced the role of measuring MSAFP levels as a screening method for NTDs. This change was initiated more about 10 years ago in some countries, which have issued national guidelines to use mid-trimester ultrasonography instead of measuring MSAFP levels as a prenatal screening method for NTDs. However, no significant changes have occurred in Korea, where second-trimester ultrasonography is routinely performed with high-quality equipment. We aimed to provide information regarding the importance of changing the screening method for NTDs from MSAFP measurement to ultrasonography, and to detail methods of implementing mid-trimester ultrasonography for screening purposes. We also share our experience of operating a prenatal diagnostic program for NTDs without using MSAFP for more than 15 years

Chemotherapeutic Candidate Inducing Immunological Death of Human Tumor Cell Lines

The immunological death induction by EY-6 on the human tumor cell lines was screened. Human colon carcinoma (HCT15, HCT116), gastric carcinoma (MKN74, SNU668), and myeloma (KMS20, KMS26, KMS34) cells were died by EY-6 treatment with dose-dependent manner. CRT expression, a typical marker for the immunological death, was increased on the EY-6-treated colorectal and gastric cancer cells. Interestingly, the effects on the myeloma cell lines were complicated showing cell line dependent differential modulation. Cytokine secretion from the EY-6 treated tumor cells were dose and cell-dependent. IFN-γ and IL-12 secretion was increased in the treated cells (200% to over 1000% of non-treated control), except HCT116, SNU668 and KMS26 cells which their secretion was declined by EY-6. Data suggest the potential of EY-6 as a new type of immuno-chemotherapeutics inducing tumor-specific cell death. Further studies are planned to confirm the efficacy of EY-6 including in vivo study

Redox-Active Star Molecules Incorporating the 4-Benzolypyridinium Cation: Implications for the Charge Transfer Efficiency Along Branches versus Across the Perimeter in Dendrimers

We report the redox properties of four star systems incorporating the 4-benzoyl-N-alkylpyridinium cation; the redox potential varies along the branches, but remains constant at fixed radii. Voltammetric analysis (cyclic voltammetry and differential pulse voltammetry) shows that only two of the three redox-active centers in the perimeter are electrochemically accessible during potential sweeps as slow as 20 mV/s and as fast as 10 V/s. On the contrary, both redox centers of a branch are accessible electrochemically within the same time frame. These results are discussed in terms of slow through-space charge transfer and the globular 3-D folding of the molecules

Incidence and Risk Factors Associated with Superior Mesenteric Artery Syndrome following Surgical Correction of Scoliosis

STUDY DESIGN: Retrospective study. PURPOSE: To more accurately determine the incidence and clarify risk factors. OVERVIEW OF LITERATURE: Superior mesenteric artery syndrome is one of the possible complications following correctional operation for scoliosis. However, when preliminary symptoms are vague, the diagnosis of superior mesenteric artery syndrome may be easily missed. METHODS: We conducted a retrospective study using clinical data from 118 patients (43 men and 75 women) who underwent correctional operations for scoliosis between September 2001 and August 2007. The mean patient age was 15.9 years (range 9~24 years). The risk factors under scrutiny were the patient body mass index (BMI), change in Cobb's angle, and trunk length. RESULTS: The incidence of subjects confirmed to have obstruction was 2.5%. However, the rate increased to 7.6% with the inclusion of the 6 subjects who only showed clinical symptoms of obstruction without confirmative study. The BMI for the asymptomatic and symptomatic groups were 18.4+/-3.4 and 14.6+/-3, respectively. The change in Cobb's angle for the asymptomatic and symptomatic groups were 24.8+/-13.6 degrees and 23.4+/-9.1 degrees , respectively. The change in trunk length for the asymptomatic and symptomatic groups were 2.3+/-2.1 cm and 4.5+/-4.8 cm, respectively. Differences in Cobb's angle and the change in trunk length between the two groups did not reach statistical significance, although there was a greater increase in trunk length for the symptomatic group than for the asymptomatic group. CONCLUSIONS: Our study shows that the incidence of superior mesenteric artery syndrome may be greater than the previously accepted rate of 4.7%. Therefore, in the face of any early signs or symptoms of superior mesenteric artery syndrome, prompt recognition and treatment are necessaryope