Proteome Analysis of a Catalase-deficient Isogenic Mutant of Helicobacter pylori 26695

Helicobacter pylori, a gram-negative bacterium, is a causative agent of gastroduodenal diseases of human. Human immune system produces harmful reactive oxygen species to kill this bacterium that locates the microaerophilic mucous layer. H. pylori harbors various antioxidant enzymes including SodB, KatA and AhpC to protect the oxygen toxicity. We removed the catalase gene (katA) from H. pylori 26695 genome, and the change of profile of the gene expression of the mutant was analyzed by high resolution 2-DE followed by matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), tandem MS and microarray analysis. Eleven and 37 genes were upregulated and downregulated in the mutant respectively, either transcriptionally or translationally. Expression level of pfr and hp1588 that were decreased on protein level in the mutant was confirmed by RT-PCR analysis.

Comparison of Gene Expression Patterns between Helicobacter pylor 26695 and its Superoxide Dismutase Isogenic Mutant

Helicobacter pylori, a causative agent of gastroduodenal diseases, is a Gram-negative microaerophilic bacterium. Although H. pylori locates in the microaerophilic mucous layer, the bacteria would come into contact harmful reactive oxygen species generated by host immune system. It has been reported that H. pylori harbors various defense mechanisms which can protect bacterial cells from oxygen exposure. The change of the gene expression profile of sodB-negative isogenic mutant of H. pylori 26695 was analyzed by high resolution 2-DE followed by matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and tandem MS and microarray analysis. Eighteen genes and 41 genes were upregulated and downregulated respectively, either transcriptionally or translationally. Expression levels of three genes including trxB, yxjE and ribE that were changed both on a mRNA level and on a protein level were confirmed by RT-PCR analysis. However, change of expression levels of other major antioxidants such as KatA, AhpC and NapA were not detected, which means Sod is regulated by different way from that of KatA and AhpC. Mutant study of other antioxidant proteins may give us better understanding for the regulation of stress response in H. pylori.

A Case Report of Methimazole-Induced Acute Liver Failure Successfully Treated with Liver Transplantation / 대한내과학회지

Antithyroid drugs inhibit the synthesis and excretion of thyroid hormone from the thyroid gland. Propylthiouracil (PTU) and methimazole are well known as antithyroid drugs. In 2011, the American Thyroid Association and American Association of Clinical Endocrinologists published management guidelines for hyperthyroidism and other causes of thyrotoxicosis, and recommended methimazole as the first-choice antithyroid drug for the treatment of hyperthyroidism. Lower hepatotoxicity is an advantage of methimazole. Fulminant hepatitis rarely occurs in methimazole users, and spontaneous recovery is expected even if it does occur. We describe a rare case of acute liver failure after methimazole intake in a 60-year-old man who underwent liver transplantation.

Experience in 102 Non-melanoma Skin Cancer with Mohs Micrographic Surgery / 대한피부과학회지

BACKGROUND: Mohs micrographic surgery (MMS) is a specialized type of minimal marginal surgery, which offers superior cure rates to other options in the treatment of non-melanoma skin cancers. OBJECTIVE: The aim of this study is to investigate the clinical characteristics and recurrence rate of the non-melanoma skin cancer and to estimate the safeties of MMS. METHODS: We retrospectively evaluated 102 patients who had been diagnosed with non-melanoma skin cancer and treated by MMS from May 2008 to September 2011. RESULTS: The most common cancer was basal cell carcinoma (63.72%), followed by squamous cell carcinoma (31.38%). The local flap was the most common method to repair surgical defects (74.5%), followed by primary closure (17.64%). Cervical lymph node metastasis was confirmed in 1 case (0.98%). Recurrence after MMS was observed in 1 of 102 patients (0.98%). CONCLUSION: MMS is a safe method for the treatment of non-melanoma skin cancer.

Identification of Helicobacter pylori Strain 51 Major Outer Membrane Proteins by Quadrupole Time of Flight Mass Spectrometry

As part of an initial inquiry into the function of the outer membrane proteins (OMPs) of Helicobacter pylori Korean strain 51, we have conducted an extensive proteome analysis via quadrupole time of flight (Q-TOF) mass spectrometry (MS). Fifty one OMPs of H. pylori were purified using sarcosine and resolved via two-dimensional electrophoresis with immobilized pH gradient strips. The most abundant proteins were observed in the alkaline pI regions (6.0~11.0) at molecular masses between 10~100 KDa. Here, 15 spots were identified, representing 9 types of genes (KHP0852, KHP0853, KHP1353, KHP1017, KHP0172, KHP0076, KHP0617, KHP1069, KHP0614) from the sarcosin-insoluble fraction of H. pylori 51. These may be employed in the characterization of the OMPs of H. pylori 51, which will help to identify new potential target proteins for vaccine development and drug therapy.

Isotretinoin Treatment for the Recalcitrant Acne on a Patient with Apert Syndrome / 대한피부과학회지

Apert syndrome is a rare genetic craniofacial disorder associated with premature fusion of multiple cranial sutures and syndactyly. It is caused by mutation in the fibroblast growth factor receptor 2 genes. The clinical findings are characterized by the typical dysmorphic facial features, including hypertelorism, proptosis and cleft palate. Most of these patients have widespread, severe acne involving the face, chest, back and forearms during puberty and the acne is unresponsive to conventional treatments. We report here on a patient with the recalcitrant acne of Apert syndrome and the patient had failed to respond to conventional acne treatment, but the acne was successfully treated with isotretinoin

Proteomic analysis of Helicobacter pylori J99 Outer Membrane Protein by Tandem Mass Spectrometry

The protein identity of sarcosine-insoluble outer membrane proteins (OMPs) of Helicobacter pylori J99 was determined with the basic study of understanding the function of proteins. A sarcosine-insoluble OMPs was resolved by two-dimensional electrophoresis with immobilized pH gradient strips. The most abundant proteins were shown in the alkaline pI regions (6.0~11.0) with molecular masses of 10 to 100 kDa. We have performed an extensive proteome analysis by quadrupole time of flight (Q-TOF) mass spectrometry (MS). Here, of 50 spots processed, 42 spots were identified, which represented 16 genes and we newly detected 8 kinds of proteins (JHP0119, JHP0388, JHP1046, JHP1405, JHP0073, JHP0551, JHP1382, JHP0552) from the sarcosin-insoluble fraction of H. pylori J99. Those may be used to elucidate the characterization of the OMPs of H. pylori J99, which will help identify new potential target proteins for vaccine development and drug therapy.

Proteomic Analysis of Helicobacter pylori Whole Cell Proteins using the Narrow Range IPG Strips

It has been reported that most of Helicobacter pylori proteome components appear so crowded in the region of pH 4.5~8.0 that a lot of them were inseparable in 2-DE using the broad range IPG strip. Therefore, inseparable protein spots in 2-DE profiles have to be apart from each other for improving the protein identification. Here, we attempt to examine the usability of the narrow range IPG strips for separating close spots in the broad range IPG strip at proteomic analysis of H. pylori. The whole cell proteins of H. pylori strain 26695 were separated by narrow range IPG strips (pI 3.9~5.1, 4.7~5.9, 5.5~6.7, and 6.3~8.3, respectively), followed by SDS-PAGE, and visualized by silver staining, showing that the distances between spots were widened and the total number of detectable spots was increased. Resolved protein spots were identified by the peptide fingerprinting using MALDI-TOF-MS. As a result, 87 expressed proteins were identified by the peptide fingerprinting. Of them, 23 proteins, including hydrogenase expression/formation protein, purine-binding chemotaxis protein, and ribosomal protein S6, have not been reported in the previous proteome studies of H. pylori. Thus, these results demonstrate that the high complexity proteome components could be effectively separated using the narrow range IPG strips, which might be helpful to strengthen the contents of the master protein map of the H. pylori reference strain.

Comparison of Proteome Component of Helicobacter pylori in Different Atmospheric CO2 Concentration

Helicobacter pylori is a spiral, slow growing gram-negative microaerophilic bacterium. It has been shown to be the etiological agent of gastroduodenal diseases, such as chronic gastritis, gastric and duodenal ulcers, and gastric cancer. General culture condition of H. pylori is 5% O2, 10% CO2 and 100% humid atmosphere. We have compared proliferation protein expression profile of H. pylori incubated under normal microaerophilic (10% CO2) and environment stress (4% CO2, 18% CO2) conditions. H. pylori cultured under environment stress displayed coccoid morphology and timedependent decrease in proliferation. We have further compared the protein expression profiles of H. pylori under normal growing and environment stress conditions by a global proteomic analysis, which includes high-resolution 2-DE followed by matrix-assisted laser desorption/ionization time of flight and nanoelectrospray/tandem mass spectrometry. In total, 42 protein spots were found to be up- or down-regulated by more than 2-fold under environment stress conditions. Of the 42 protein spots processed, 27 spots were identified; they represented 19 genes, including 2 kinds of hypothetical proteins.

Expression of the 14-3-3 sigma Protein and Methylation Status of the 14-3-3 sigma gene in Biliary Neoplasms

BACKGROUND: The 14-3-3 sigma (sigma) protein has a negative regulatory role in the cell cycle progression of the. Down-regulation or overexpression of the 14-3-3 sigma protein has been reported in various human cancers. METHODS: Immunohistochemistry for the 14-3-3 sigma protein was performed in non-neoplastic bile duct cells, intraductal papillary neoplasms of the liver (IPNL), mass-forming intrahepatic cholangiocarcinomas (ICC) and non-papillary extrahepatic cholangiocarcinomas (ECC). We investigated the methylation status of the 14-3-3 sigma gene in 45 cases of these 3 tumor groups. RESULTS: The non-neoplastic bile duct cells demonstrated negative or weakly positive cytoplasmic immunoreactivity for the 14-3-3 sigma protein and no methylation of the 14-3-3 sigma gene. Overexpression as well as negative immunoreactivity associated with hypermethylation of the 14-3-3 sigma protein was observed in 16 (69.6%) of 23 cases of IPNL, in 21 (63.6%) of 33 cases of mass-forming ICC and in 27 (71.1%) of 38 cases of non-papillary ECC. Negative immunoreactivity was increased in the invasive IPNL (4/6, 66.7%), as well as in the poorly differentiated cases of mass-forming ICC (8/12, 66.7%) and the non-papillary ECC (5/8, 62.5%). CONCLUSIONS: The similar rates for the abnormal expression of the 14-3-3 sigma protein among the three groups of biliary neoplasms indicate its general association with biliary carcinogenesis. Furthermore, the loss of the 14-3-3 sigma protein may be involved in the tumor progression and differentiation in the biliary carcinogenesis.

A Case of Acquired Digital Arteriovenous Malformation / 대한피부과학회지

Arteriovenous malformation (AVM) consists of an abnormal connection between artery and vein without any interposed capillary bed, and is usually congenital. However, acquired AVM is quite rare, and is usually caused by injury, which induces direct arteriovenous shunting. Acquired digital AVM is a distinct entity, which was first described by Kadono et al. in 2000, and was thought to result from shunts between an artery and a vein in a finger tip. We report a case of acquired digital AVM in a 44-year-old women. The patient showed punctate hyperketatotic ectasia on the pulp of the right thumb. Histological finding showed dilated venous and arterial vessels in upper and lower dermis, and shunts between arteriols and venules.

A Case of Microvenular Hemangioma / 대한피부과학회지

Microvenular hemangioma is an uncommon acquired vascular tumor of young to middle-aged adults of both genders. It usually presents as a small, solitary, purple to red papule or plaque on the extremities or trunk. Histologically, the tumor is composed of small branching venules with collapsed lumina and conspicuous pericytes infiltrating the full thickness of the reticular dermis. Because of histological similarity to a Kaposi's sarcoma, the awareness of microvenular hemangioma is important. We report a case of microvenular hemangioma in a 9-year-old girl.

A Case of Congenital Triangular Alopecia in Infancy / 대한피부과학회지

Congenital triangular alopecia is an uncommon nonscarring form of alopecia. It presents with isolated alopecic patch at the frontotemporal area of the scalp. Although previously considered congenital, this condition usually is discovered after 2 years of age and, more recently, is thought to be acquired. We report a case of congenital triangular alopecia in a 12 month old female who had the lesion since birth.

Therapeutic Effect of Topical Diphenylcyclopropenone (DPCP) for the Treatment of Extensive Alopecia areata / 대한피부과학회지

BACKGROUND: Alopecia areata (AA) is a common dermatologic disorder and the course is so variable that some patients undergo spontaneous remission and others undergo total hair loss. There is no clearly superior therapy for the treatment of alopecia areata. Currently, topical immunotherapy with DPCP represents the most accepted therapeutic modality for the treatment of extensive alopecia areata, but their response rates have varied in the literature. OBJECTIVE: We evaluated the efficacy, prognostic factors, and side effects of DPCP in the treatment of extensive AA. METHOD: Thirty nine patients with extensive AA (>50 % scalp hair loss), treated for at least 6 months at the Department of Dermatology of Inha University Hospital between March 2000 and April 2003 participated in the study. After sensitization with 1% DPCP, progressively higher concentrations beginning from 0.001% were applied weekly onto the entire scalp. The primary study end point, i.e. clinically significant regrowth with DPCP therapy, was defined as a cosmetically acceptable response (as judged by the patient) or significant regrowth resulting in greater than 90% of the scalp being covered with terminal hair (as determined by the investigators) RESULT: A clinically significant regrowth was obtained in 91.3% of the patients with 50% to 99% AA and 50.0% with alopecia totalis/universalis. The overall clinically significant regrowth rate was 74% (29 of 39 patients). Variables associated with clinically significant regrowth were the beginning age of DPCP therapy and the extent of AA. Relapse was observed in 56.3% of the patients who achieved significant hair regrowth after 6 months of follow-up. The Clinically significant adverse effects observed were eczematous reaction with blistering, swelling of cervical lymph nodes, urticaria, and erythema multiforme. CONCLUSION: Treatment with DPCP for extensive AA is very effective. Response of AA patients to DPCP treatment is affected by the beginning age of DPDP therapy and the extent of AA.

Two Cases of Eosinophilic Pustular Folliculitis in Infancy / 대한피부과학회지

Eosinophilic pustular folliculitis(EPF) is characterized by recurrent crops of pruritic follicular papules and pustules that occur mainly on the face, trunk, and extremities of young men. EPF in infancy is a rare disorder which reveals the distinctive clinical features consisting of self-limited recurrent papules, pustules, and vesicles appearing in corps, involving mainly or exclusively the scalp. We describe two patients with EPF in infancy. Both patients had self-limited recurrent crops of papules and pustules affecting the scalp only.

A Cases of Erythema Elevatum Diutinum / 대한피부과학회지

Erythema elevatum diutinum(EED) is a rare chronic skin disease characterized by red, purple, and yellowish papules, plaques, and nodules that are usually distributed acrally and symmetrically over extensor surface. It is characterized histologically by a leukocytoclastic vasculitis. Although no treatment has shown a complete response, dapsone is reported to be effective to EED. We report a rare severe case of EED with good response to dapsone treatment.

A Case of Polycythemia Vera with Splinter Hemorrhages

Once splinter hemorrhage can be considered as a pathognomonic sign of subacute bacterial endocarditis. But it can also be associated with a variety of systemic disorders that increase capillary fragility or primary nail bed involvement in dermatologic disorders. The cause of splinter hemorrhage can usually be established by careful history and physical examination. We report a case of 33-year-old man with splinter hemorrhages, who had polycythemia vera.

A Case of Sneddon's Syndrome / 대한피부과학회지

Sneddon's syndrome is an infrequent neurocutaneous disorder of unknown origin. It is characterized by the combination of livedo reticularis and cerebrovascular accident. We present a 57-year-old male patient with livedo reticularis and cerebrovascular accident. Magnetic resonance imaging of the head showed a sign of acute focal infarctions in the right cerebellar hemisphere and right vermis. He had netlike patterned, mottled bluish discoloration on both legs. Histopathologic finding revealed elongation and fusion of rete ridges and mild thickening of dermal capillaries.

Two Cases of Combined Liver-kidney Transplantation in Patients with End Stage Renal Diseases Associated with Hepatitis B Related Liver Cirrhosis / 대한신장학회잡지

We here report two cases of combined liver-kidney transplantation in patients with both end stage renal disease and hepatitis B related liver cirrhosis. The first case was a 55-year-old man with hepatitis B related liver cirrhosis and chronic renal failure, who received cadaveric liver and kidney transplantation. Immunosuppressants were cyclosporine, prednisolone, and mycophenolate mofetil. Clinical course was uneventful except for hemolytic anemia due to alloimmunization that occurs after ABO-mismatched solid organ transplantation. Hemoglobin level became stable after plasmapheresis. His renal and hepatic function is maintained up to the present time. The second case was a 42-year-old man with nephrotic syndrome and liver cirrhosis. The patient underwent living related-combined liver-kidney transplantation. Donors were his son and brother. Blood type of the patient and donors were identical and the result of HLA crossmatch was negative. On the 14th postoperative day, stenosis at anastomotic site of hepatic artery was detected. After balloon angioplasty hepatic function was normalized. At 8 months after the transplantation, the patient is stable without adverse events.

Expression of Transforming Growth Factor Alpha mRNA in Livers of Patients with Chronic Viral Hepatitis and Hepatocellular Carcinoma / 대한간학회지

BACKGROUND/AIMS: Transforming growth factor alpha (TGFalpha) is an important autocrine growth factor of hepatocytes. We were to evaluate the roles of TGFalpha in chronic viral hepatitis (CVH) and hepatocellular carcinoma (HCC). METHODS: We measured the amounts of TGFalpha mRNA in liver tissues from 18 patients with HCC, 31 with CVH and 7 normal controls. 'Hot start' reverse transcription polymerase chain reaction (RT-PCR) using oligo-dT and specific primers detected TGFalpha mRNA in total cellular RNA extracted from liver tissues. The levels of TGFalpha mRNA were determined by the end point titers of serial two-fold dilutions of cDNA. The amounts of HBV-RNA in livers of patients with chronic hepatitis B were also measured by Northern blot hybridization. RESULTS: TGFalpha mRNA levels were significantly higher in HCC than CVH and normal controls; those of CVH were also elevated compared to normal controls. They were overexpressed in the nontumorous livers surrounding HCC compared to CVH, although lower than HCC tissues. They were higher in chronic hepatitis B than in chronic hepatitis C. They were not correlated with serum alanine aminotransferase (ALT) or HBV-RNA levels in liver tissues (in cases of chronic hepatitis B). However, the expression of TGFalpha mRNA tended to be higher in the livers of patients with serum AFP levels raised. CONCLUSION: Overexpression of TGFalpha mRNA in liver seems to be associated with the regeneration of hepatocytes rather than hepatic necrosis or viral replication. Also, it may be closely related to the development or growth of HCC, especially in the livers of chronic hepatitis B.