Application of Mac-2 binding protein glycosylation isomer as a non-invasive biomarker for probing liver disease.

Liver disease remains a major critical challenge in Thailand due to viral hepatitis. Clinical management requires close monitoring of liver fibrosis severity. Non-invasive testing is an attractive method for probing of disease progression. Mac-2 binding protein glycosylation isomer (M2BPGi) is a novel serum marker for fibrosis staging. The current study evaluates the marker among healthy donors and hepatitis C (HCV) patients. 100 HCV subjects were evaluated by liver biopsy. These patients had varying fibrosis severity based on METAVIR scores. Healthy donors were confirmed based on normal liver functions tests. Comparisons of M2BPGi levels among different study groups were performed and the effectiveness was evaluated using receiver operating characteristics (ROC) curves. Using liver biopsy as the reference standard, median M2BPGi levels in HCV cases were 0.74, 1.38 and 2.88 COI for F0-1, F2 and > F3 cases respectively. In healthy donors, the baseline values ranged 0.1-0.24 COI and statistically lower than liver disease cases profiled using M2BPGi. ROC analysis demonstrated superior results for M2BPGi levels among diseased populations and healthy controls. AUROC was determined at 0.983. Comparing with other non-invasive tests, M2BPGi showed a positive linear trend that indicated a strong match to existing methodologies. M2BPGi addresses a critical need in the management of liver disease by providing straightforward means to probe fibrosis severity. In this study, we found significant differences between hepatitis C and healthy subjects and established the background level in healthy donors.

A COMPLICATED CASE OF STRONGYLOIDIASIS PRESENTING WITH INTESTINAL LYMPHADENOPATHY OBSTRUCTION: MOLECULAR IDENTIFICATION.

Strongyloides stercoralis is an intestinal nematode, which can cause complications in immune-compromised hosts. We present a rare case of intestinal obstruction due to mesenteric lymphadenopathy, a complication due to strongyloidiasis, developing in a male subject chronically receiving corticosteroid for pemphigus vulgaris. DNA was extracted from biopsied lymph nodes containing nematode larvae and PCR amplified using primers specific for S. stercoralis 18S rDNA. Nucleotide sequence of the amplicon showed identity with that of S. stercoralis deposited in GenBank. To the best of our knowledge, this is the first report of a diagnosis of strongyloidiasis from biopsied samples using molecular techniques.

Modulation of antibody responses against Gnathostoma spinigerum in mice immunized with crude antigen formulated in CpG oligonucleotide and montanide ISA720.

This study aimed to investigate the antibody responses in mice immunized with Gnathostoma spinigerum crude antigen (GsAg) incorporated with the combined adjuvant, a synthetic oligonucleotide containing unmethylated CpG motif (CpG ODN 1826) and a stable water in oil emulsion (Montanide ISA720). Mice immunized with GsAg and combined adjuvant produced all antibody classes and subclasses to GsAg except IgA. IgG2a/2b/3 but not IgG1 subclasses were enhanced by immunization with CpG ODN 1826 when compared with the control groups immunized with non-CpG ODN and Montanide ISA or only with Montanide ISA, suggesting a biased induction of a Th1-type response by CpG ODN. After challenge infection with live G. spinigerum larvae, the levels of IgG2a/2b/3 antibody subclasses decreased immediately and continuously, while the IgG1 subclass remained at high levels. This also corresponded to a continuous decrease of the IgG2a/IgG1 ratio after infection. Only IgM and IgG1 antibodies, but not IgG2a/2b/3, were significantly produced in adjuvant control groups after infection. These findings suggest that G. spinigerum infection potently induces a Th2-type biased response.

PCR-RFLP and antimicrobial susceptibility profiles of Helicobacter pylori isolated from antrum and corpus of dyspeptic patients in Thailand.

The study determined the genetic heterogeneity of Helicobacter pylori isolates from antrum and corpus of the same dyspeptic patients in a Thai population and determined the relationship between the antimicrobial susceptibility (AS) profile (antibiogram) and PCR-restriction fragment length polymorphism (PCR-RFLP) pattern. One hundred and nineteen H. pylori isolates comprising 7 single and 56 paired antrum and corpus isolates obtained by gastric biopsy from 160 dyspeptic patients were analyzed. For PCR-RFLP, the 820 bp amplicon of ureC was digested with Sau3AI and HhaI, which revealed 16 (A-Q) and 19 (a- s) different PCR-RFLP patterns after Sau3AI and HhaI digestion, respectively. Combination of the restriction enzyme digestion patterns resulted in 35 distinct RFLP types. Among the 56 paired isolates, 47 were infected with H. pylori having the same AS and PCR-RFLP profiles, 7 with different AS profiles but the same PCR-RFLP profiles and 2 with different PCR-RFLP profiles but the same AS profiles. No patient was infected with H. pylori different in both PCR-RFLP and AS profiles. The results indicate that the majority of the paired H. pylori isolates displayed identical AS profile and PCR-RFLP patterns suggesting that most patients were infected with a single strain. Some patients could have been infected with single strains that were different in the AS profiles.

Melioidosis pericarditis mimicking tuberculous pericarditis.

Cases of melioidosis (N = 2) and tuberculous pericarditis (N = 33) during 1996-2006 were reviewed. Clinical presentations were similar, but pericardial pathological findings were not. Nine of 12 patients with melioidosis required pericardectomy. In areas where these diseases are endemic, pericardial fluid culture and pericardial biopsy can differentiate between melioidosis and tuberculosis.

Glutathione S-transferase (GSTM1 and GSTT1) polymorphisms in cervical cancer in Northeastern Thailand.

To evaluate the relationships between genetic polymorphisms of the GSTs (GSTM1 and GSTT1) and cervical cancer, the null genotype of each gene was studied in squamous cell cervical cancer (SCCA) patients (n=90) and controls (n=94) in Northeast Thailand. The prevalence of the GSTM1-null genotype in the controls and SCCA patients was 59.6% and 60.0%, respectively, whereas those of the GSTT1-null genotype in the control and SCCA patients was 40.4% and 46.7%, respectively. Neither of the GST-null genotypes increased the risk for SCCA (p>0.05); however, the combination of the GSTM-1 and GSTT1-null genotypes showed a non-significant trend for an increased risk for developing cervical cancer with an adjusted OR of 2.7 (95%CI=0.8-9.0, p=0.10). Genetic polymorphisms of GSTM1 and GSTT1 were not significant risk factors for cervical cancer in either tobacco-smokers or non-smokers. A different contribution of the GST genotype to cancer risk may be attributed to a different, as yet undefined, property of the enzymes.

Mycotic aneurysm in Northeast Thailand: the importance of Burkholderia pseudomallei as a causative pathogen.

Forty cases of mycotic aneurysm that occurred during the period 1993-2007 were reviewed. The most common causative pathogen was Burkholderia pseudomallei (17 cases; 42.5%). Postoperative complications and bacteremia were significantly more common among patients with mycotic aneurysm due to B. pseudomallei than they were among patients with mycotic aneurysm that was not attributable to B. pseudomallei. In a region in which melioidosis is endemic, empirical antimicrobial therapy for suspected mycotic aneurysm should cover B. pseudomallei.

Visual inspection with acetic acid test qualities in a secondary setting.

AIM: To evaluate the visual inspection using acetic acid (VIA) test qualities in a secondary (follow-up) setting, 1 year after cryotherapy treatment performed as part of the Safety, Acceptability and Feasibility demonstration project designed to evaluate the safety, acceptability and feasibility of VIA. An immediate offer of cryotherapy was made to those who test positive and are eligible for treatment. METHODS: At 1 year after cryotherapy, 648 women received both a secondary (follow-up) VIA test by nurse-providers, and colposcopy with biopsy, if indicated, by trained physician colposcopists at a referral hospital. All pathologic specimens were sent for examination by a single pathologist. RESULTS: VIA nurse-providers assessed 42 of the 648 women (6.5%) referred as abnormal (i.e. they tested positive or were suspected of having cancer). Among the 42, the final colposcopic-based diagnosis was HSIL or higher in three cases (7.1%), of which two were HSIL and one was adenocarcinoma. Of the 606 VIA negative women, the colposcopic-based diagnosis was HSIL in only two cases (0.3%). CONCLUSIONS: The VIA test qualities in this setting were: a positive rate of 6.5%, a sensitivity of 60%, a specificity of 93.9%, a positive predictive value of 7.1%, a negative predictive value of 99.7% and an accuracy of 93.7%. These results are comparable to those of Pap smear in most settings.

Fine needle aspiration in a malignant lymphoepithelial lesion: a case report.

BACKGROUND: A primary malignant lymphoepithelial lesion (MLEL) of the salivary gland is a rare tumor. Though histologic morphology of this lesion is well documented, the cytologic findings regarding fine needle aspiration (FNA) are not yet well described. CASE: A 56-year-old Thai woman from Udonthanee was admitted to Srinagarind Hospital with a 2-year history of a painless mass in the left parotid gland. FNA of the left parotid mass was performed, and findings suggestive of malignancy were discovered. A wide local excision was performed 1 year later, and MLEL with metastasis to the regional lymph nodes was diagnosed by histopathology. CONCLUSION: We report our first experience of cytologic findings from FNA of histologically diagnosed MLEL. The aspirate comprised groups of cohesive and isolated malignant epithelials with a background of numerous lymphocytes. Cytotechnologists should be reminded of this rare lesion when confronted with a lymphocytic background from either benign or malignant lesions.

The endoscopic-pathologic findings in intestinal capillariais: a case report.

A 27-year-old Thai man presented with chronic watery diarrhea for 2 years. The diagnosis of capillariasis was made by enteroscopy after negative repeated stool tests. Here, the authors reported the first case of abnormal endoscopic finding of intestinal capillariasis. It showed segmental erythematous and swelling of proximal jejunal mucosa with an area of superficial erosion covered by exudates. The parasitic eggs were identified in jejunal content and worms were identified in jejunal mucosa. He was successfully treated with albendazole.

Bisphosphonate-related osteonecrosis of the jaws (ONJ): a report of two cases.

Osteonecrosis of thejaw (ONJ) is strongly associated with the use of aminobisphosphonates. Herein, the authors report two cases of ONJ after intravenous bisphosphonate therapy including clinical presentations, X-ray, and pathological findings. Since there is no definite treatment for ONJ, the focus should be on prevention with a dental evaluation for all patients before starting bisphosphonates.

Sensitivity and specificity of an in-house rapid urease test for detecting Helicobacter pylori infection on gastric biopsy.

We developed an in-house rapid urease test (iRUT) and evaluated the efficacy and the agreement of the iRUT and the cRUT compared with culture and histology for the detection of H. pylori infection. Five iRUT media were tested with H. pylori isolates and other bacteria. The most suitable iRUT medium was further evaluated for detection of H. pylori infection. Gastric biopsies from 120 patients were diagnosed by culture, iRUT, cRUT and histology. The results of the iRUT and cRUT were read at 30 minutes, 1 hour and up to 24 hours. A true positive result was either the culture or both the RUT (cRUT or iRUT) and the histological examination being positive. The sensitivity and specificity of the iRUT result at 30 minutes, 1 hour and up to 24 hours were 77.1% and 100%, 77.6% and 100%, and 94.1% and 94.2%, respectively. Values for the same parameters of cRUT were 87.5% and 100%, 89.8% and 100%, and 100% and 94.2%, respectively. The agreement between the iRUT and cRUT was very good (kappa values > or = 0.82). Our results indicate that the iRUT is a-sensitive, specific and cost effective test. It can be appropriately applied for detecting H. pylori infection in gastric biopsy specimens.

Selected risk factors, human papillomavirus infection and the p53 codon 72 polymorphism in patients with squamous intraepithelial lesions in northeastern Thailand.

Risk factors for cervical squamous intraepithelial lesions (SIL) including human papillomavirus (HPV) infection and the p53 codon 72 polymorphism were investigated in a case-control study with 103 cases and 105 controls in Northeastern Thailand. Increased risk for SIL was observed for age at menarche (odds ratio (OR) = 2.2; p< 0.005), age at the first sexual intercourse (OR=2.4; p< 0.05), number of sexual partners (OR=2.7; p< 0.005) and partners' smoking history (OR=2.3-3.2; p< 0.01). Prevalence of malignant type of HPV infection in the control and SIL groups was 18.1% and 60.2%, respectively. HPV infection significantly increased risk for SIL 6.8-fold (p< 0.001). HPV-16 infection was the commonest (31 out of 62 carriers) in SIL patients and highly associated with risk. The p53 codon 72 polymorphism was not identified as a genetic risk for SIL in this study, as demonstrated in Thai cervical cancer. Therefore, to prevent cervical neoplasia or HPV infection, inclusion of knowledge on sexual behavior and effects of smoking into public health programs is important and, at the same time, a nation-wide screening scheme for cervical abnormalities including HPV-typing is a high priority in Thailand.

Enzyme-linked immunosorbent assay for serodiagnosis of Helicobacter pylori in dyspeptic patients and volunteer blood donors.

Helicobacter pylori, an important etiological agent in the development of gastritis, peptic ulcer and gastric carcinoma, can be detected by the enzyme-linked immunosorbent assay (ELISA). Our objectives were: (1) to evaluate the efficacy of a commercial ELISA kit (Pyloriset EIA-G III) in sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy for diagnosis of H. pylori infection in Thai dyspeptic patients in Khon Kaen Thailand; and (2) to examine the seroprevalence of H. pylori among blood donors at Srinagarind Hospital's Blood Bank, Khon Kaen University, by the commercial ELISA. Gastric biopsies obtained from 137 dyspeptic patients were diagnosed by culture, rapid urease test (RUT) and histology. Serum samples from the same dyspeptic patients and 100 healthy blood donors were assayed using the commercial ELISA. H. pylori infection in dyspeptic patients was considered positive when the culture or both RUT and histology were positive. Using a cut-off value at a titer of 20 U/ml (as recommended by the manufacturer), we found the commercial ELISA kit had a sensitivity of 93.3%, specificity of 75.3%, PPV of 74.7%, NPV of 93.5% and accuracy of 83.2%. The overall H. pylori seroprevalence in the healthy blood donors was 57%. Of the 100 healthy blood donors, 39 (60.9%) of the males and 18 (50.0%) of the females were seropositive.

Human papillomavirus genotypes and the p53 codon 72 polymorphism in cervical cancer of Northeastern Thailand.

Human papillomavirus (HPV) infection including sub-strain identification was studied in patients with squamous cell cervical cancer (SCCA) in Northeastern Thailand. Subjects were 90 cases of SCCA and 100 healthy controls. Prevalence of high-risk group of HPV infection in the controls and the SCCA patients were 13.0% and 86.7%, respectively. The HPV infection significantly increased the risk for cervical cancer 43.5-fold (95% confidential interval: 17.5-110.6; P <0.00001). Among HPV carrier patients with SCCA (n = 78), HPV-16 was also prominent (70.5%) followed by HPV-18 (23.1%). There was no statistical difference in the subtype distribution between the SCCA and the control groups. There was no significant association between genotype distribution of the p53 codon 72 polymorphism and HPV infection. HPV infection was confirmed as a critical risk factor for cervical cancer development in Northeast Thailand. Since polymorphism of the p53 itself as well as in combination with HPV infection may not be a genetic risk for cervical cancer, much attention should be paid to other risk factors such as sexual behavior and smoking.

Rituximab combined with CHOP for successful treatment of aggressive recurrent, pediatric B-cell large cell non-Hodgkin's lymphoma.

This report is the first to describe the successful treatment of a 14-year-old boy with aggressive recurrent, CD20-positive, B-cell large cell non-Hodgkin's lymphoma. The patient responded to three 4-week courses of rituximab (MabThera) given every 6 months and six cycles of CHOP given every 3 weeks in addition to a modified BFM 86 protocol. Transient neutropenia and lymphopenia occurred but with no clinical significance. The boy has been disease-free for the last 48 months (after 64 months of follow-up); his organ functions are normal. Rituximab and CHOP in addition to chemotherapy may be an alternative treatment for aggressive recurrent, pediatric CD20-positive B-cell large cell non-Hodgkin's lymphoma if highly intensive chemotherapy and stem cell transplantation are not available.

Clinical value of biliary alkaline phosphatase in non-jaundiced cholangiocarcinoma.

PURPOSE: Measurement of the activities of alkaline phosphatase isoenzyme has been used for the identification and monitoring of diseases associated with the isoenzyme. Biliary alkaline phosphatase (BALP), an isoform of liver-ALP, has been found in the serum of patients with biliary obstruction and metastatic liver cancer. This study compared the BALP isoform in the serum of patients with cholangiocarcinoma (CCA) with that of non-jaundiced benign hepatobiliary disease, other cancers, and healthy persons. METHODS: ALP isoforms were separated using cellulose acetate electrophoresis and the activity was demonstrated using indolyl blue reagent. RESULTS: The BALP isoform was demonstrated in 65% of CCA patients independently of jaundice condition or histological grading of the tumor. The level of serum BALP in non-jaundiced CCA was significantly lower than that of jaundiced CCA, and not correlated with serum bilirubin. No BALP was detected in healthy persons. In the patients with high serum ALP (> 147U/l), BALP can differentiate non-jaundiced CCA patients from other non-jaundiced carcinoma patients with 85% sensitivity, 79% specificity, and positive and negative predictive values of 81% and 83%, respectively. CONCLUSION: The demonstration of serum BALP, in particular in non-jaundiced patients with high serum ALP, may indicate the presence of tumor in the bile duct.