The glycoprotein hormone family consists of follicle-stimulating hormone (FSH; GTH1), luteinizing hormone (LH; GTH2), and thyroid-stimulating hormone (TSH), which are secreted by the pituitary gland in all mammalian species, and chorionic gonadotropin, which is secreted by placental trophoblast cells in primates and equids. These hormones consist of non-covalently associated α-, β- subunits. Within a species, the amino acid sequence of α-subunit is identical across all glycoprotein hormones and is encoded by a single gene. The αβ dimer is the active form of the hormone, and biological specificity is conferred by the β-subunit. Both of α and β subunit of eel FSH has two N-glycosylation sites (α-subunit: Asn56 and Asn79; β -subunit: Asn5 and Asn22, respectively). In the present study, we constructed deglycosylated mutants at single and double sites in each subunits of eel FSH for identification of Asn linked oligosaccharides' biological role. Mutant cDANs were cloned into pcDNA3 expression vector and transiently transfected into CHO suspension cells. The quantity of rec-eelFSHs were quantified by sandwich ELISA system, using monoclonal antibodies produced in our lab. The wild type rec-FSH protein was detected at the predicted molecular weight of 34 kDa by western blot. The molecular weight of deglycosylated mutants at single site decreased with about 4 kDa and of mutants at double sites decreased with 8 kDa. After PNGase treatment in the rec-eel FSH proteins, molecular weight also decreased to 7-8 kDa. We generated stably parental cell lines, engineered to express a β-arrestin 2EA fusion protein, expressing eel FSHR and C-terminal deleted mutant. 2 out of 5 receptor cells each were selected by G-418 and we tested these cell lines in a receptor functionality using PathHunter β arrestin assay (DiscoverX). Follicle stimulating hormone acts through binding to its specific receptor. Binding of ligand to the receptor activates the adenosine 3',5'-cyclic monophosphate (cAMP) pathway (McFarland et al., 1989; Ji and Ji, 1991a; Rose, 1998) and the inositol 1phosphate (IP1) the second messenger systems. After stimulation of eelFSH receptor stably transfected Parental CHO cells with FSH wild type and mutant hormones as a ligand, production of cAMP and IP-1 were evaluated (Cisbio). cAMP IC-50 values by eelFSHwt; αΔ56; αΔ79; αΔ56_79; and βΔ5 were 33.1; 1154.7; 22; 410 and 311.9 ng/ml, respectively. IP-1 IC-50 values by eelFSHwt; αΔ56; αΔ79; αΔ56_79 and βΔ5 were 6.8; 7.1; 4.4; 3.8 and 10.2 ng/ml, respectively too. The cAMP activation was greatly decreased in the αΔ56αmutant. Thus, the site of α56 oligosaccharide in the eel plays an pivotal role for the cAMP stimulation using eel FSH receptor cell lines. In the IP-one assay, the activity in the αΔ56 and βΔ5 mutants was a little decreased than the wt. The biological roles of N-linked oligosaccharides in GPCR internalization are going to be estimated by measuring β arrestin recruitment system.

In this study, we developed and validated microanalysis methods for the determination of linear alkylbenzenesulfonate (LAS), sodium lauryl sulfate (SLS), and alpha olefin sulfonate (AOS). The conditions for the analysis of the surfactants using HPLC with FLD, RID, and ELSD detectors were investigated. The methods were validated by determining the linearity, limits of detection (LODs), limits of quantification (LOQs), recovery, precision, and accuracy. LAS analysis by FLD revealed calibration curves that were linear in the range of 10-200 mg/L for an LAS mixture. The calibration curves for C10-C13 had correlation coefficients of 0.995, 0.997, 0.996, and 0.997, respectively. SLS analysis using RID generated a linear calibration curve in the range of 10-300 mg/L. The calibration curve for SLS C12 had a correlation coefficient of 0.9994. AOS analysis using ELSD resulted in a correlation coefficient of 0.9940. For LAS, the LODs and LOQs were 0.09-0.56 and 0.30-1.87 mg/L, respectively. For SLS C12, the LOD and LOQ were 0.07 and 2.33 mg/L, respectively. For AOS C14, the LOD and LOQ were 16.55 and 21.83 mg/L, respectively. The recoveries were 97.17-98.84% for LAS C10-C14, 97.94% for SLS C12, and 96.11% for AOS C14. The established methods provide acceptable precision and accuracy. Our methods could be useful for the detection of anionic surfactants in dishwashing detergents.

Breast specific gamma imaging (BSGI) is a nuclear medicine breast imaging technique. The sensitivity of BSGI is unaffected by post-operative scars or implants, and thus, BSGI is helpful for the differential diagnosis of postoperative recurrence. Here, the authors report a case of diffuse chest wall recurrence on the side of radical mastectomy that was not detected by BSGI, but was detected using other nuclear medicine imaging techniques.

We compared the results of early follow-up of F-18 FDG PET/CT and MRI performed within one month after radiation therapy for cervix cancer patients. We conducted a retrospective review of the clinical data of cervix cancer patients whose PET/CT and pelvic MRI performed at staging and within one month from the end of RTx. SUVmax on PET/CT and size on MRI of the primary tumor were analyzed. We compared %change of SUVmax and size between staging and follow-up. A total of 27 patients were enrolled. At staging, larger tumor showed high SUVmax. At follow-up, no significant correlation was observed between size and SUVmax. In 77.8% of patients, changes in SUVmax were well correlated with changes in size. No correlation was observed between % change and value at staging in both SUVmax and size. Except for six patients who showed significant FDG uptake without evidence of a mass on MRI, % changes of size and SUVmax were well correlated. Metabolic change can be accessible on early follow-up PET/CT at±1 month from the end of the RTx of cervix cancer. However, careful interpretation of PET/CT is needed due to possible radiation-induced hypermetabolism even without a definite mass on MRI.

A 56 year-old women with a history of breast cancer underwent Tc-99m Hydroxymethylene Diphosphonate (HDP) bone scintigraphy at an annual follow-up, and abnormal focal uptake was observed in the right upper abdomen. However, subsequent single photon emission computed tomography/computed tomography (SPECT/CT) successfully delineated the uptake in a normal gallbladder. It was concluded that the abnormality had been caused by unusual Tc-99m HDP excretion and not by a metastatic lesion. Follow-up studies confirmed this diagnosis. This case demonstrates the usefulness of SPECT/CT in patients with unusual gallbladder uptake by bone scintigraphy.

Sea Buckthorn (Hippophae rhamnoides L.) has been used in traditional medicine for the treatment of cough, indigestion, circulatory problems and pain. The associated anti-inflammatory effect of this agent is achieved via the inhibition of Nf-kB signaling, a property that has been demonstrated to effectively control the symptoms of various skin disorders, including atopic dermatitis. Accordingly, the purpose of this study was to assess the efficacy of Sea Buckthorn in reducing the production of lipopolysaccharide (LPS) activated nitric oxide (NO) by inhibiting the Nf-kB pathway, as measured by the symptoms of atopic dermatitis (AD) occurring secondarily to inflammation and immune dysregulation. Our data demonstrate that Sea Buckthorn significantly decreased the LPS-induced production of NO (p〈0.001). Atopic dermatitis was induced by repeated application of 2,4-dinitrochlorobenzene to the dorsal skin of mice. Topical application of 5% Sea Buckthorn extract improved the symptoms of AD, specifically reducing disease severity scores, scratching behaviors and epidermal thickness. When compared to the control group, animals treated with Sea Buckthorn exhibited increased serum IL-12 levels and decreased serum TNF-α, IL-4 and IL-5 levels. Such a modulation of biphasic T-helper (Th)1/Th2 cytokines may result in a reduction in serum IgE levels. Our findings suggest that mechanism of action of Sea Buckthorn in the treatment of AD is associated with a marked anti-inflammatory effect as well as an inhibition of Th2-mediated IgE overproduction via the modulation of biphasic Th1/Th2 cytokines. Such results suggest that topical Sea Buckthorn extract may prove to be a novel therapy for AD symptoms with few side effects.