Association between 24-hour combined multichannel intraluminal impedance-pH monitoring and symptoms or quality of life in patients with laryngopharyngeal reflux.

OBJECTIVE: To evaluate the association between the parameters of 24-hour multichannel intraluminal impedance (MII)-pH monitoring and the symptoms or quality of life (QoL) in laryngopharyngeal reflux (LPR) patients. DESIGN: Prospective cohort study without controls. SETTING: University teaching hospital. METHODS: Forty-five LPR patients were selected from subjects who underwent 24-hour MII-pH monitoring and were diagnosed with LPR from September 2014 to May 2015. Reflux Symptom Index (RSI), Health-related Quality of Life (HRQoL), Short Form 12 (SF-12) Survey questionnaires were surveyed. Spearman's correlation was used to analyse the association between the symptoms or QoL and 24-hour MII-pH monitoring. RESULTS: Most parameters in 24-hour MII-pH monitoring showed weak or no correlation with RSI, HRQoL and SF-12. Only number of non-acid reflux events that reached the larynx and pharynx (LPR-non-acid) and number of total reflux events that reached the larynx and pharynx (LPR-total) parameters showed strong correlation with heartburn in RSI (R = 0.520, P < 0.001, R = 0.478, P = 0.001, respectively). Multiple regression analysis showed that there was only one significant regression coefficient between LPR-non-acid and voice/hoarseness portion of HRQoL (b = 1.719, P = 0.022). CONCLUSION: Most parameters of 24-hour MII-pH monitoring did not reflect subjective symptoms or QoL in patients with LPR.

Effect of postoperative proton pump inhibitor therapy on voice outcomes following phonomicrosurgery for vocal fold polyp: a randomized controlled study.

OBJECTIVE: To determine the effect of a postoperative proton pump inhibitor (PPI) on voice outcomes after phonomicrosurgery in patients with vocal fold polyp. STUDY DESIGN: This is a prospective, randomized controlled study. SETTINGS: This study was carried out in a tertiary care referral medical centre. PARTICIPANTS: A total of 48 patients underwent phonomicrosurgery for vocal fold polyps. After surgery, patients were randomized to the PPI group (lansoprazole 15 mg twice daily for 2 months) and the non-PPI group. MAIN OUTCOME MEASURES: Voice handicap index (VHI) and perceptual and acoustic voice analysis were evaluated at baseline and 2 months after surgery. RESULTS: Among 48 enrolled patients, a total of 42 patients [non-PPI group (n = 23), PPI group (n = 19)] completed the study. The VHI, perceptual and most acoustic parameters significantly improved in both groups after surgery. However, there was no significant difference in the per cent of change in those parameters. CONCLUSION: Postoperative PPI treatment did not significantly influence voice outcomes after phonomicrosurgery in patients with vocal fold polyp.

Evaluation of salivary function in patients with burning mouth syndrome.

OBJECTIVES: To investigate salivary function in patients with primary burning mouth syndrome (BMS) compared with control and to evaluate salivary hypofunction using salivary gland scintigraphy (SGS). METHODS: A total of 33 patients with primary BMS and 30 control subjects were enrolled in our study. The severity of the pain and the burning sensation on a 10-cm visual analog scale (VAS) and the Oral Health Impact Profile-14 (OHIP-14) were assessed. Unstimulated and stimulated salivary flow rates (SFRs) were measured. (99m) Tc pertechnetate SGS was used to evaluate salivary gland function. RESULTS: Unstimulated SFR in patients with BMS was significantly lower than that in the control group (0.11 ± 0.15 vs 0.21 ± 0.16 ml min(-1) , P = 0.014). There was no significant difference in stimulated SFR between the two groups. The VAS scores for oral pain and burning sensation, the total OHIP-14 score, and salivary gland function by salivary scintigraphy were not significantly different between BMS patients with normal flow rate and hyposalivation. CONCLUSIONS: Patients with primary BMS exhibited a significant decrease in unstimulated SFR compared with control group. In addition, we could not find any difference in salivary gland function between BMS patients with or without hyposalivation.

Application of IFN-γ releasing assay for the diagnosis of erythema induratum of Bazin.

BACKGROUND: Erythema induratum of Bazin (EIB) is regarded to be a hypersensitive reaction to the concomitant tuberculosis. Recently, interferon-γ releasing assay (IGRA) has been focused as a promising tool in the diagnosis of latent tuberculosis. However, there has been no large scale study to investigate the usefulness of IGRA in the diagnosis of EIB. OBJECTIVES: To evaluate the diagnostic performance for the detection of EIB. METHODS: We retrospectively reviewed medical records of all patients with EIB, in the Department of Dermatology, at the Seoul National University Hospital, between April 2009 and September 2011. We analysed clinicopathological features, responses to IGRA and the treatment courses. In addition, we compared positive rate of IGRA in patients with other diseases during the same period. RESULTS: All of the 22 patients demonstrated a positive response to IGRA (100%) and showed a good response to anti-tuberculosis treatment. In contrast, positive rate was 63.64% and 66.67% in patients with psoriasis and other vasculitis respectively. We observed complete resolution of skin lesions in 14 patients. Partial resolution was attained in one patient and the other seven patients are currently on the medication and are showing good responses. CONCLUSION: We verified that IGRA has an excellent diagnostic performance in EIB, through this observational study. It is strongly suggested that if EIB is clinicopathologically suspected, IGRA should be performed.

Long-pulsed Nd : YAG laser: does it give clinical benefit on the treatment of resistant telangiectasia?

BACKGROUND: Long-pulsed Nd : YAG laser has been used in treating larger and deep-seated leg veins. OBJECTIVE: This study investigated the effectiveness and safety of long-pulsed Nd : YAG laser to treat alae nasae and nasal tip telangiectasia. METHODS: Twelve patients were evaluated in a prospective IRB approved study. They had a history of previous unsatisfactory treatments with pulsed dye laser and/or intense pulsed light for their telangiectases on alae nasae and tip (4-12 times, average 5.8 times). All patients underwent a single treatment session using long-pulsed Nd : YAG laser. Photographic images were taken. At 12-week follow-up, two independent physicians evaluated the percentage of vessels cleared, and patients were asked to rate their satisfaction with the procedure. RESULTS: Five men and seven women were enrolled (aged 43 ± 5.8 years). Total clearance of vessels was 78.3%. The number of vessels in diameter of 0.1 mm was reduced by 61.1% and that of vessels in diameter of 0.2-0.3 mm decreased by 92.2% on the average at 12-week follow-up. Eleven of 12 patients were very satisfied with the clinical results. One patient rated as 'satisfied' due to hyperpigmentation after the treatment, which improved at 12-week follow-up. CONCLUSION: Long-pulsed Nd : YAG laser can be considered as another effective and safe treatment modality for stubborn telangeictasia even on face, if applied cautiously.

Environmental assessment of water, sediment and plants in the Mankyeong River, ROK.

This study was carried out to evaluate water quality, sediment and plant vegetation in eight tributaries of the Mankyeong River for enhancement of natural purification. Among the tributaries, the Iksancheon water had the highest concentration of BOD, T-N and NH4-N due to inflow of swine wastes from the livestock district. The Yucheon water had the highest level of electrical conductivity and SO4(2-) due to inflow of mis-treated wastewater from industrial districts. The Tabcheon had generally similar concentrations of nitrogen and phosphate to that of the upstream of the Mankyeong River: agricultural activity along the Tabcheon appeared to have little negative influence to the water quality. Among various sediments, concentration of organic matter, nitrogen and phosphate were high in the Iksancheon and the Yucheon due to the livestock wastes and industrial wastes. There were 282 species of plants during summer with 43 aquatic plants, 57 hydrophytes, 178 waterside plants and 4 terrestrial plants. Some plant resources were recommended due to much absorption of nitrogen and phosphate for enhancement of natural purification. C. demersum and H. verticillata were recommended in the submerged aquatic plants, H. dubia, N. indica and N. subinteperrimum in the floating leaf aquatic plants, P. communis, Z. latifolia and T. orientalis in the emerged aquatic plants, C. scutata and P. distichum in the waterside plants.

HLA-DR expression in B-lymphocytes in vitro is not suppressed by the absence of exogenous antigens.

The proper loading of exogenous peptide antigens affects the transport and cell surface expression of MHC class II molecules. In the present study, the goal was to determine to what extent this step determines the cell surface expression level of MHC class II molecules, such as the HLA-DR. EBV-transformed B-cells, were cultured either in a serum- and protein-free medium, or in a medium that contained different concentrations of exogenous antigens. Using HLA-DR-specific antibodies, the induction of the MHC class II expression was observed in cells that were cultured under serum-and protein-free conditions, when compared to those cultured with exogenous protein antigens. This upregulation was completely suppressed to the normal level by the addition of a high concentration of hen egg lysozyme to the serum- and protein-free medium. This indicates that exogenous proteins regulate the HLA-DR expression. To further examine whether this modulation is controlled at the transcription level, the expression of the HLA-DR beta-chain mRNA was analyzed by reverse transcription-PCR and Northern blots. The same levels of HLA-DRB mRNA were detectable in both culture conditions, indicating that the present observation is dependent on some regulatory mechanisms at the post-transcriptional level. This might include a different pathway for trafficking of HLA-DR molecules to the cell surface, since peptide-binding assays revealed that a high proportion of cell surface HLA-DR molecules under the serum- and protein-free condition were transported to the cell surface without associated peptide antigens.

Determination of rat leptin activity in vitro using a novel luciferase reporter assay.

Leptin is small cytokine-like protein that is involved in appetite and body weight regulation. Due to increased interest in using leptin as an anti-obesity reagent, recombinant forms of leptin have been produced for several species, including humans, mice, rats, pigs, dogs, sheep etc. The biological activities of such recombinant proteins were determined using various in vitro or in vivo systems; however so far, no specific assay system for rat leptin is available. Since rats are representative animal models in obesity research, the establishment of a biological assay system for determining rat leptin activity has been eagerly awaited. This study describes the generation of such a system using chinese hamster ovary (CHO)-cells that were transfected with the long form of the rat leptin receptor isoform, OB-Rb, whereby a signal transduces and activators of transcription-sensitive luciferase reporter system is further employed to quantify the leptin-mediated signals. This system is the first rat-specific leptin bioassay system that has been reported. It is expected that this assay will be used to further quantify and determine leptin activity from various biological fluids and sources.

Recombinant expression of biologically active rat leptin in Escherichia coli.

Leptin is a 16-kDa nonglycosylated hormone that is produced in mature adipocytes and which acts primarily in the hypothalamus to reduce food intake and body weight. While the rat is a representative laboratory animal model in obesity research, so far recombinant rat leptin was not available. In the present study, rat leptin was recombinantly expressed in Escherichia coli and purified in a bioactive form to provide a further tool for the analysis of leptin functions in rats. Leptin cDNA was cloned by RT-PCR from total RNA of SD rat adipocytes, and overexpression was achieved by subcloning the leptin cDNA into the pET-29a vector, which enabled the recombinant expression of rat leptin as an S-peptide-tagged fusion protein. Since the fusion proteins were expressed in inclusion bodies, after purification of the insoluble fraction, leptin proteins were refolded by sequential dialysis into physiological buffers. The biological activity of this recombinant protein was confirmed in proliferation assays using leptin-sensitive rat insulinoma cells as well as a newly developed leptin-sensitive luciferase assay system. The specific binding of the S-tagged leptin to leptin-receptor-expressing cells was further shown by flow cytometry using fluorescence-conjugated S-proteins.

Detection of pET-vector encoded, recombinant S-tagged proteins using the monoclonal antibody ATOM-2.

The 15-meric S-tag is a truncated form of the S-peptide, which builds together with the 103 amino acid large S-protein the whole ribonuclease S-protein. Its small size and excessive solubility have made the S-tag an excellent fusion partner in the production of recombinant proteins, and a large variety of applications have been reported using the S-tag as a carrier. While S-tagged proteins were mostly detected and analyzed so far by use of their affinity to S-proteins, monoclonal antibodies (MAbs) for this tag have been not available. The generation of antibodies specific for S-tagged proteins is expected to broaden the range of applications of such S-tag fused recombinant proteins, and in this context, a novel MAb termed ATOM-2 was generated that specifically binds S-tagged proteins, which have been expressed using pET-vectors. Antigen specificity of ATOM-2 was confirmed in Western blot and enzyme-linked immunoadsorbent assay analysis, and using a series of amino acid deletion mutants, the binding epitope of ATOM-2 was precisely mapped. This showed that ATOM-2 recognizes the C-terminal part of the 15-meric S-tag in context with a few residues of vector encoded sequences. The core sequence for ATOM-2 binding epitope is "His-Met-Asp-Ser-Pro-Asp-Leu-Gly-Thr," which is present in all pET-expression vectors encoding S-tag fusion proteins. Because the ATOM-2 binding region does not overlap with the S-protein binding sequence, a convenient tool is provided for the simultaneous or alternative detection, purification, and analysis of recombinant S-tagged proteins to conventional S-proteins.

Transcriptional coregulators of the nuclear receptor superfamily: coactivators and corepressors.

Nuclear receptors, many of which undergo a major conformational change upon binding specific ligand, belong to a superfamily of proteins that bind to specific DNA sequences and control gene transcription. They regulate the assembly of a transcriptional preinitiation complex at the promoter of target genes and modulate their expression in response to ligand. In particular, nuclear receptors repress or stimulate transcription by recruiting corepressor or coactivator proteins, in addition to directly contacting the basal transcription machinery. In this review, we discuss recent progress in studies of these transcriptional coregulators of nuclear receptors.

Two distinct nuclear receptor-interaction domains and CREB-binding protein-dependent transactivation function of activating signal cointegrator-2.

ASC-2 is a recently isolated transcriptional cointegrator molecule, which is amplified in human cancers and stimulates transactivation by nuclear receptors, AP-1, nuclear factor kappaB (NFkappaB), serum response factor (SRF), and numerous other transcription factors. ASC-2 contained two nuclear receptor-interaction domains, both of which are dependent on the integrity of their core LXXLL sequences. Surprisingly, the C-terminal LXXLL motif specifically interacted with oxysterol receptor LXRss, whereas the N-terminal motif bound a broad range of nuclear receptors. These interactions appeared to be essential because a specific subregion of ASC-2 including the N- or C-terminal LXXLL motif acted as a potent dominant negative mutant with transactivation by appropriate nuclear receptors. In addition, the autonomous transactivation domain (AD) of ASC-2 was found to consist of three separable subregions; i.e. AD1, AD2, and AD3. In particular, AD2 and AD3 were binding sites for CREB binding protein (CBP), and CBP-neutralizing E1A repressed the autonomous transactivation function of ASC-2. Furthermore, the receptor transactivation was not enhanced by ASC-2 in the presence of E1A and significantly impaired by overexpressed AD2. From these results, we concluded that ASC-2 directly binds to nuclear receptors and recruits CBP to mediate the nuclear receptor transactivation in vivo.

Inactivation of p16INK4a in primary tumors and cell lines of head and neck squamous cell carcinoma.

Inactivation of the p16INK4a gene by mutation and deletion is common in head and neck squamous cell carcinoma (HNSCC). The present study demonstrates that hypermethylation of the 5' CpG islands can serve as an alternative mechanism for the inactivation of the p16INK4a gene in this tumor. We studied 11 HNSCC cell lines and 17 oral squamous cell carcinoma (OSCC) primary tumors for p16INK4a gene status by protein/mRNA and DNA genetic/epigenetic analyses to determine the incidence of its inactivation. Our study indicates that: (1) inactivation of p16 protein is frequent in HNSCC cell lines (6/11, 54.5%) and OSCC primary tumors (15/17, 88.2%), (2) inactivation of p16INK4a protein is commonly associated with the presence of gene alteration such as mutation, homozygous deletion and especially aberrant methylation, and (3) genomic sequencing of bisulfite-modified DNA shows that the carcinoma develops a heterogeneous pattern of hypermethylation.

Detection of cellular receptors specific for the hepatitis B virus preS surface protein on cell lines of extrahepatic origin.

Hepatitis B virus infection is primarily mediated by the interaction of the preS region of the viral envelope protein with its still unknown cellular receptor. Using recombinantly expressed preS proteins, the distribution of preS-binding receptors on cell lines from extrahepatic origins was determined by immunofluorescence and flow cytometry. In contrast to human liver cell lines, most cell lines from extrahepatic origins did not bind preS proteins. Nevertheless, exceptions were found in the bone marrow-derived cell line, KG-1, and the osteogenic sarcoma cell line SaOS-2, as well as in the previously reported EBV-transformed B-cell line, Wa. To determine the biochemical nature of these receptors, Wa-cells were cell surface biotinylated and the preS-binding receptors were isolated by immunoprecipitation. A specific band with a molecular weight of approximately 30 kDa was identified in a SDS-polyacrylamide gel, which further characterization is expected to provide clues regarding the infection mechanism of HBV in hepatic- and extra-hepatic cells.

Genomic organization and tissue-specific expression of rat urocortin.

The genomic organization of rat urocortin was determined using both cDNA and liver genomic DNA as templates for polymerase chain reactions. A single intron of 261 bp was found upstream to the ATG start codon, and the whole urocortin coding sequence was shown to be contained in a single exon. Relying on these results, oligonucleotide primers were designed, which can differentiate genomic DNA contamination from cDNA-derived signals in a reverse transcription-polymerase chain reaction. Tissue screening for urocortin expression revealed that the mid-brain is the major site of urocortin mRNA expression, and that other peripheral organs, including lymphoid tissues and peripheral blood lymphocytes, do not produce urocortin.

Activating protein-1, nuclear factor-kappaB, and serum response factor as novel target molecules of the cancer-amplified transcription coactivator ASC-2.

ASC-2 was recently discovered as a cancer-amplified transcription coactivator molecule of nuclear receptors, which interacts with multifunctional transcription integrators steroid receptor coactivator-1 (SRC-1) and CREB-binding protein (CBP)/p300. Herein, we report the identification of three mitogenic transcription factors as novel target molecules of ASC-2. First, the C-terminal transactivation domain of serum response factor (SRF) was identified among a series of ASC-2-interacting proteins from the yeast two-hybrid screening. Second, ASC-2 specifically interacted with the activating protein-1 (AP-1) components c-Jun and c-Fos as well as the nuclear factor-kappaB (NFkappaB) components p50 and p65, as demonstrated by the glutathione S-transferase pull-down assays as well as the yeast two-hybrid tests. In cotransfection of mammalian cells, ASC-2 potentiated transactivations by SRF, AP-1, and NFkappaB in a dose-dependent manner, either alone or in conjunction with SRC-1 and p300. In addition, ASC-2 efficiently relieved the previously described transrepression between nuclear receptors and either AP-1 or NFkappaB. Overall, these results suggest that the nuclear receptor coactivator ASC-2 also mediates transactivations by SRF, AP-1, and NFkappaB, which may contribute to the putative, ASC-2-mediated tumorigenesis.