Jongkolnee Settakorn

T‐type calcium channel as a portal of iron uptake into cardiomyocytes of beta‐thalassemic mice

Objectives: Iron‐overload condition can be found in β‐thalassemic patients with regular blood transfusion, leading to iron deposition in various organs including the heart. Elevated cardiac iron causes iron‐overload cardiomyopathy, a condition that provokes mortality because of heart failure in patients with thalassemia. Previous studies demonstrated that myocardial iron uptake may occur via L‐type calcium channels (LTCCs). However, direct evidence regarding the claimed pathway in thalassemic cardiomyocytes has never been investigated. Methods: Hearts from genetic‐altered β‐thalassemic mice and adult wild‐type mice were used for cultured ventricular cardiomyocytes. Blockers for LTCC, T‐type calcium channel (TTCC), transferrin receptor1 (TfR1), and divalent metal transporter1 (DMT1) were used, and quantification of cellular iron uptake under various iron loading conditions was performed by Calcein‐AM fluorescence assay. Microarray analysis was performed to investigate gene expressions in the hearts of these mice. Results: This study demonstrated that iron uptake under iron‐overload conditions in the cultured ventricular myocytes of thalassemic mice was greater than that of wild‐type cells (P < 0.01). TTCC blocker, efonidipine, and an iron chelator, deferoxamine, could prevent iron uptake into cultured cardiomyocytes, whereas blockers of TfR1, DMT1, and LTCC could not. Microarray analysis from thalassemic hearts demonstrated highly up‐regulated genes of TTCC, zinc transporter, and transferrin receptor2. Conclusions: Our findings indicated that iron uptake mechanisms in cultured thalassemic cardiomyocytes are mainly mediated by TTCC, suggesting that TTCC is the important pathway for iron uptake in this cultured thalassemic cardiomyocyte model.

Chondroprotective and anti-inflammatory effects of sesamin

Osteoarthritis (OA) is a major disability of elderly people. Sesamin is the main compound in Sesamun indicum Linn., and it has an anti-inflammatory effect by specifically inhibiting Δ5-desaturase in polyunsaturated fatty acid biosynthesis. The chondroprotective effects of sesamin were thus studied in a porcine cartilage explant induced with interleukin-1beta (IL-1β) and in a papain-induced osteoarthritis rat model. With the porcine cartilage explant, IL-1β induced release of sulfated-glycosaminoglycan (s-GAG) and hydroxyproline release, and this induction was significantly inhibited by sesamin. This ability to inhibit these processes might be due to its ability to decrease expression of MMP-1, -3 and -13, which can degrade both PGs and type II collagen, both at the mRNA and protein levels. Interestingly, activation of MMP-3 might also be inhibited by sesamin. Moreover, in human articular chondrocytes (HACs), some pathways of IL-1β signal transduction were inhibited by sesamin: p38 and JNK. In the papain-induced OA rat model, sesamin treatment reversed the following pathological changes in OA cartilage: reduced disorganization of chondrocytes in cartilage, increased cartilage thickness, and decreased type II collagen and PGs loss. Sesamin alone might increase formation of type II collagen and PGs in the cartilage tissue of control rats. These results demonstrate that sesamin efficiently suppressed the pathological processes in an OA model. Thus, sesamin could be a potential therapeutic strategy for treatment of OA.

HPV genotyping in cervical cancer in Northern Thailand: Adapting the linear array HPV assay for use on paraffin-embedded tissue

OBJECTIVES The aims of this study were to determine the prevalence of HPV infection and distribution of HPV genotypes in Northern Thai women and thereby estimate the benefit of administering the HPV vaccine in the population. METHODS Formaldehyde-fixed, paraffin-embedded samples of invasive squamous cell carcinoma from 99 patients were tested for HPV genotypes using the Linear Array HPV Genotyping Test. RESULTS HPV was detected in 96/99 (96.9%) cases. Seventy-five (78.1%) cases were single infections and 21 (21.9%) multiple. HPV16 and HPV18 were the most common subtypes, detected in 62/96 (64.4%) cases. HPV52 and HPV58 infections were found in 17/96 (17.7%) cases. Co-infection always involved HPV16. The most common co-infection was HPV16 and 52 (7 cases) but never HPV16 and 18. CONCLUSIONS Although the prevalence of HPV infection in cervical cancer of Northern Thai women is comparable to the other regions worldwide, the distribution of HPV subtypes differs with lower frequencies of HPV16 and 18, and higher frequencies of HPV52 and 58. Moreover, multiple infections are common. The vaccine against HPV16 and HPV18 can be estimated to prevent approximately two thirds of the cervical cancer cases in Northern Thailand. Although designed for use on unfixed tissue, this study shows that the Linear Array HPV Genotyping Test can be successfully used for HPV genotyping on paraffin-embedded archival tissue. This methodology also provides a means for retrospective studies on serial samples for a greater understanding of HPV genotypes, co-infections, and relationship to cervical cancer.

Mucinous tumor of low malignant potential ("borderline" or "atypical proliferative" tumor) of the ovary: a study of 171 cases with the assessment of intraepithelial carcinoma and microinvasion.

Mucinous tumors of the ovary are a continuing source of controversy in the field of gynecologic pathology. We examined a series of 171 intestinal-type mucinous tumors of low malignant potential (“borderline” or “atypical proliferative” tumors) to clarify the clinical significance of intraepithelial carcinoma (IECA) and microinvasion (area ⩽10 mm2). The diagnosis of IECA was based on the presence of marked nuclear atypia (grade 3). Stromal microinvasion was classified as low grade and high grade (with nuclear grade 3). IECA was observed in 67 of 171 cases (39.2%). Microinvasion was identified in 31 (18.1%) cases, low grade in 22 (12.9%) cases, and high grade in 9 (5.3%) cases. Follow-up status was known in 144 cases and tumor recurrence was observed in 6 patients (4.2%). The risk factors for recurrence included International Federation of Gynecology and Obstetrics stage ≥IC (P=0.002), microinvasion (P=0.013), age less than 45 years (P=0.032), and IECA (P=0.042). The amount of IECA ≥10% was also associated with the risk of recurrence (P=0.007). Among tumors with microinvasion, there was no significant association between the clinicopathologic variables and recurrence. When considering tumors with stage ≥IC, tumor recurrence was significantly associated with IECA ≥10% (P=0.031) and age less than 45 years (P=0.047). It is important that mucinous tumors of low malignant potential should be staged and be optimally sampled for pathologic examination to document the status of the external surface or peritoneal involvement and to identify the worst degree of epithelial proliferation. Tumor stage ≥IC, IECA ≥10%, microinvasion, and age less than 45 years were the features that were associated with tumor recurrence. The study results also support the use of nuclear grade 3 as the sole criterion of IECA.

Accuracy of Frozen-section Diagnosis of Ovarian Mucinous Tumors

Objective The objective of the study was to evaluate the diagnostic accuracy of intraoperative frozen sections of ovarian mucinous tumors and to identify the features associated with an inaccurate diagnosis. Methods Cases of ovarian mucinous tumors (benign, low malignant potential [LMP] or borderline, primary malignant, and metastatic) diagnosed by frozen section or final histology were recruited. Frozen-section diagnoses were compared with the final histologic diagnoses. Possible variables associated with diagnostic discrepancy were analyzed. Results A comparison of the diagnoses was done in 195 cases (102 benign, 61 LMP, 18 primary malignant, and 14 metastatic). Diagnostic agreement was observed in 164 cases (84.1%) and discrepancy in 31 cases (15.9%). The sensitivity of frozen-section diagnosis was low in LMP (67.2%) and malignant tumors (55.6%). The specificity was the lowest in the benign category (78.5%). The positive predictive values of all categories were less than 90% (range, 83.3%–85.7%). Diagnostic discrepancy was associated with tumor size of greater than 13 cm (P = 0.019) and the number of frozen sections of 4 or more (P = 0.035). However, in a multivariate analysis, there was no independent predictor of diagnostic discrepancy. The number of frozen sections 4 or more was strongly associated with tumor size of greater than 13 cm (P = 0.004). Conclusions The sensitivity of frozen-section diagnosis of LMP and malignant mucinous tumors was low. The inaccuracy of a frozen-section diagnosis of ovarian mucinous tumors may be related to a tumor size of greater than 13 cm. Increasing the number of intraoperative samples over 3 sections per case may not effectively increase the accuracy of frozen-section diagnosis in mucinous tumors.

Cardiac mortality is associated with low levels of omega-3 and omega-6 fatty acids in the heart of cadavers with a history of coronary heart disease.

The benefits of omega-3 (ie, eicosapentaenoic acid and docosahexaenoic acid [DHA]) and omega-6 (ie, linoleic acid and arachidonic acid [AA]) fatty acids on reducing cardiac mortality are still debated. In this study, we tested the hypothesis that high levels of omega-3 and omega-6 fatty acids in heart tissues are associated with low cardiac mortality in Thai cadavers. One hundred fresh cadavers were examined in this study. The cause of death, history of coronary heart disease (CHD), and fish consumption habits were obtained from death certificates, cadaver medical record profiles, and a questionnaire to a person who lived with the subject before death. In each cadaver, biopsies of cardiac tissues were taken from the interventricular septum for measurement of fatty acid. Of the 100 cadavers (average age, 69 +/- 13 years), 60 were men. The frequency of fish consumption was directly associated with omega-3 and omega-6 fatty acids in heart tissues (P < .01). History of CHD and cause of death (cardiac vs noncardiac) were not significantly associated with levels of omega-3 or omega-6 fatty acids. However, in cadavers with a history of CHD, high levels of omega-3 and omega-6, particularly DHA and AA, were associated with low cardiac mortality (P < .05). Fish consumption is associated with levels of omega-3 and omega-6 fatty acids in heart tissues. Although omega-3 and omega-6 fatty acids are not associated with cardiac mortality in the overall studied population, their low levels (especially DHA and AA) in heart tissues are associated with high cardiac mortality in cadavers with a history of CHD.

HPV genotyping in neuroendocrine carcinoma of the uterine cervix in northern Thailand

To determine the distribution of HPV genotypes in cervical neuroendocrine carcinoma (NECA) in northern Thailand, and evaluate the correlation between HPV genotype and clinicopathologic features.

Cyclooxygenase-2 expression in squamous cell carcinoma of the uterine cervix is associated with lymph node metastasis.

OBJECTIVES Previous studies have indicated that cyclooxygenase-2 (COX-2) activity is related to the development and progression of cervical cancer. In this study, we evaluated the association between COX-2 expression and specific clinicopathologic features in surgically-treated squamous cell carcinoma of the uterine cervix. METHODS Immunohistochemical staining for COX-2 was performed on 196 cases of stage IB-IIA cervical squamous cell carcinoma. Results were correlated with the clinicopathologic features and disease-free survival using statistical analysis. RESULTS Expression of COX-2 was detected in 48.5% of cases. COX-2 expression was significantly associated with lymph node metastasis (p=0.045) but lacked significant correlation with tumour stage, size, histologic grade, deep stromal invasion, lymphovascular space invasion, and parametrial involvement. In multivariate analysis, only parametrial involvement and lymphovascular space invasion (LVSI) were independent predictors for lymph node metastasis (p=0.001 and 0.007, respectively). COX-2 expression was not associated with lymph node metastasis in the absence of parametrial involvement or LVSI. In the cases with LVSI, COX-2 expression was significantly associated with lymph node metastasis (p=0.03), although with marginal significance (p=0.068) in the multivariate analysis. COX-2 expression was not associated with a decrease in disease-free survival for patients overall (p=0.977). However, in patients who did not receive adjuvant treatment, COX-2 expression was significantly associated with decreased disease-free survival (p=0.008) and was a significant predictor of recurrence (p=0.014). CONCLUSIONS In this study, COX-2 expression was associated with lymph node metastasis in cervical squamous cell carcinoma, but this was linked to the presence of LVSI or parametrial involvement. This suggests that COX-2 expression may enhance lymph node metastasis after LVSI occurs. If so, immunohistochemical staining for COX-2 may provide additional prognostic information in LVSI-positive cases, in particular in patients who do not receive postoperative adjuvant treatment.

Prospective evaluation of serum biomarker levels and cartilage repair by autologous chondrocyte transplantation and subchondral drilling in a canine model

IntroductionThe purpose of this study was to evaluate serum chondroitin sulfate (CS) and hyaluronic acid (HA) levels and the capability of cartilage repair of full-thickness cartilage defects after treatment with two different fundamental surgical techniques: autologous chondrocyte transplantation (AC) and subchondral drilling (SD).MethodsA 4-mm-diameter full-thickness cartilage defect was created in each of 10 skeletally mature male outbred dogs. The dogs were randomly separated into two groups. Groups A and B were treated with AC and SD, respectively. An evaluation was made at the 24th week of the experiment. Serum was analyzed prospectively – preoperatively and at 6-week intervals – for CS and HA levels by enzyme-linked immunosorbent assay (ELISA) and ELISA-based assays, respectively.ResultsThe cartilage repair assessment score (median ± standard deviation) of group A (9.5 ± 2.5) was significantly higher than that of group B (2.5 ± 1.3) (P < 0.05). Group A also demonstrated a better quality of hyaline-like cartilage repair. Prospective analysis of serum WF6 and HA levels between the two groups did not show any significant difference. Serum WF6 levels at the 24th week of the experiment had a negative correlation (r = -0.69, P < 0.05) with the cartilage repair assessment score, whereas serum HA levels tended to correlate positively (r = 0.46, 0.1 <P < 0.05).ConclusionsAC treatment provides superior results to SD treatment, according to morphology, histology, and cartilage marker levels. AC treatment demonstrated a smoother surface, less fissure, better border integration, and a more reliable outcome of repairing cartilage. Moreover, a decreasing level of serum WF6, which correlated with good quality of the repairing tissue at the end of the follow-up period, was found predominantly in the AC group. Serum WF6 therefore should be further explored as a sensitive marker for the noninvasive therapeutic evaluation of cartilage repair procedures.

Angiostrongylus cantonensis infection mimicking a spinal cord tumor

Angiostrongylus cantonensis is the most common cause of eosinophilic meningitis and meningoencephalitis. Almost all cases are self‐limiting and are diagnosed by cerebrospinal fluid eosinophilia and enzyme‐linked immunosorbent assay; pathology reports are restricted to postmortem samples from lethal cases. We report on what we believe is the first case of A. cantonensis infection diagnosed by biopsy in a living patient. The spinal cord was biopsied because of the unusual clinical presentation of a myelopathy without meningeal symptoms, together with a mass lesion that was clinically and radiologically diagnosed as a spinal cord tumor.