Chulalongkorn Medical Journal

H4K20me3 is increased in human bladder cancer tissues and is upregulated by reactive oxygen species in bladder cancer cell lines

Sotida Kayem — 2024-10-01

Background: Previously, we demonstrated that reactive oxygen species (ROS) triggered oxidative stress,enhanced tumor aggressiveness, and induced a profound change in histone methylation in bladder cancer (BlCa) and reported H4K20me3 upregulation by ROS in hepatocellular carcinoma.

Objective: This study aimed to investigate whether H4K20me3 was upregulated in human BlCa tissues and determine if ROS could induce H4K20me3 formation in BlCa cell lines.

Methods: Immunohistochemical (IHC) staining was performed in 37 BlCa sections and 6 adjacent noncancerous tissues (controls). ROS upregulated H4K20me3 was investigated in three BlCa cell lines: UM-UC-3, VM-CUB- 1, and TCCSUP.

Results: The H4K20me3 levels in BlCa tissues increased relative to that in adjacent noncancerous tissues.The IHC score of the H4K20me3 level in BlCa tissues was significantly higher than that in noncancerous controls. H2O2(ROS representative) at 50, 100, and 200 mM significantly induced oxidative stress in UM-UC-3, VM-CUB-1, and TCCSUP cells, respectively, but did not significantly alter cell survival. Western blot and immunofluorescent staining results showed that H2O2treatment markedly increased H4K20me3 formation in all three cell lines.

Conclusion: This study demonstrated that the H4K20me3 levels in BlCa tissues obtained from Thai patients with BlCa increased compared with the levels in adjacent noncancerous tissues. Evidently, ROS upregulated H4K20me3 formation in BlCa cell lines. Perhaps, ROS induced the expression of histone methyltransferases that further caused an increase in H4K20me3 formation. The induction of tumor progression by ROS is well recognized; however, whether ROS induced BlCa progression is mediated by H4K20me3 formation remains to be elucidated.

Prevalence and factors associated with work-related musculoskeletal disorder among central sterile supply technicians in Nakhon Si Thammarat Province, Thailand

Withaya Chanchai — 2024-10-01

Background: Central sterile supply technicians (CSSTs) have a high risk of work-related musculoskeletal disorders (WMSDs), notably affecting the back.

Objective: This study aimed to examine the prevalence and factors associated with WMSDs among CSSTs in Nakhon Si Thammarat Province, Thailand.

Methods: A cross-sectional study assessed WMSDs among CSSTs in Nakhon Si Thammarat using self-administered questionnaires on demographics and symptoms. Out of 188 distributed questionnaires, 179 were completed, which evaluated the presence and severity of the disorders during sterilization in 33 hospitals.

Results: The prevalence of WMSDs in various body regions, particularly the upper back, at the time of questioning, during the past week, and over the past year was 88.2%, 79.3%, and 77.7%, respectively. Technicians with >30 years of experience showed a significantly higher risk of WMSDs (odds ratio [OR] 8.7,95% confidence interval [CI] 1.4–6.3). CSSTs with >10 years of employment showed significant associations with WMSDs (OR 7.9, 95% CI 1.3–5.0). Other factors included working day shifts (OR 19.2, 95% CI 10.2–120.5), heavy load handling (OR 20.0, 95% CI 10.8–153.6), lifting of instruments (OR 2.0, 95% CI 1.4–5.5),transporting equipment or carts (OR 3.0, 95% CI 1.9–7.3), pushing equipment or carts (OR 9.2, 95% CI 1.4–5.5),carrying instruments (OR 5.6, 95% CI 1.1–4.3), and uncomfortable postures (OR 7.7, 95% CI 1.4–7.6).

Conclusion: These findings highlight the high prevalence of WMSDs among CSSTs, particularly affecting the upper back. Factors such as prolonged employment and physically demanding tasks significantly contribute to these disorders.

Accuracy and concordance of DMIND AI application with a renowned depression assessment tool in Thai adults

Solaphat Hemrungroj — 2024-10-01

Background: In Thailand, the high prevalence of depression, particularly in rural areas with limited mental health services, poses significant challenges. Advanced technologies like digital phenotyping and artificial intelligence (AI), particularly natural language processing (NLP) and Large Language Models (LLMs), offer promising solutions by analyzing digital data to detect early signs of depression.

Objectives: This study evaluated the performance of the "Detection and Monitoring Intelligence Network of Depression (DMIND)" application, an AI-powered screening tool for detecting depression in Thai adults that analyzes behavioral data from participant responses using machine learning algorithms, such as NLP models and LLMs. The study aimed to determine the agreement of the DMIND AI model and the Thai version of the Hamilton depression rating scale (HDRS-17 Thai) in classifying depression severity.

Methods: This cross-sectional study recruited 388 participants from one tertiary care hospital and two psychiatric hospitals. Initially, participants used the DMIND application, where they were asked to answer a series of questions. Their response in the application was recorded and a pre-trained AI model predicted their depression severity. Subsequently, a trained nurse or psychologist then assessed participants using the HDRS-17 Thai to establish a baseline measure of depression severity. Statistical analysis involved comparing the depression severity classifications from the DMIND AI model with the HDRS-17 Thai. Cohen's kappa coefficient, sensitivity, specificity, and predictive values were used to evaluate the agreement between the two assessments.

Results: Our DMIND application demonstrated moderate agreement with the HDRS-17 Thai, indicating substantial consistency in depression severity classification. The tool showed high sensitivity (87.3%) and moderate specificity (59.5%), with strong negative predictive values for detecting depression.

Conclusion: The AI-powered DMIND application effectively screens for depression by analyzing digital data from participants' responses. Its moderate agreement with a traditional clinical assessment and strong diagnostic performance highlights its potential as a scalable, accessible tool for mental health management in Thailand. Integrating AI tools like the DMIND into the public health infrastructure could significantly enhance the accessibility, accuracy, and responsiveness of mental health services, particularly in underserved regions, potentially revolutionizing the management and treatment of depression across the country.

Histone 3 and 4 acetylation patterns in nodular goiters and well-differentiated thyroid tumors compared with normal thyroid tissue

Nishal Chhetri — 2024-10-01

Background: Histone acetylation is a well-recognized epigenetic process involved in various cancer development pathways. However, thus far, its role in thyroid tumors and nodular goiter has been barely investigated.

Objectives: This study aimed to assess histone modifications (hypoacetylation or hyperacetylation) in various thyroid lesions, including papillary thyroid carcinoma (PTC), follicular thyroid carcinoma (FTC), follicular adenoma (FA), and nodular goiter, through immunohistochemistry with anti-acetyl histone 3 (H3K9/ K14ac) and 4 (H4K5, 8, 12, and 16ac) antibodies, compared with their normal counterparts to elucidate their role in the development of these lesions.

Methods: Formalin-fixed paraffin-embedded sections of surgically resected PTC, FA, FTC, and nodular goiter samples were collected from the archives of the Department of Pathology, Faculty of Medicine, Chulalongkorn University, from 2016 to 2018. These sections were then stained with anti-acetyl histone 3 (H3K9/K14ac) and 4 (H4K5, 8, 12, and 16ac) antibodies. Aperio ImageScope was used to automatically score the intensity and proportion of immunostaining of the lesions and their normal thyroid tissue counterparts.

Results: A total of 97 malignant thyroid lesions, including (28 FA, 50 PTC, and 19 FTC samples), and 50 nodular goiters were evaluated. The deacetylation of both anti-acetyl histone 3 (H3K9/K14ac) and 4 (H4K5, 8, 12, and 16ac) antibodies was detected in nodular goiter (P = 0.0016 and P < 0.0001, respectively) in comparison with their normal counterparts. However, the difference in the acetylation status of FTC, PTC, and FA were not significant compared with that of their normal counterparts (P > 0.05 in all cases).

Conclusion: For the first time, this study demonstrates the H3 and H4 deacetylation of nodular goiters compared with their normal tissue counterparts. In contrast, these epigenetic events are not found in welldifferentiated thyroid neoplasms (FA, FTC, and PTC).

Retrospective analysis of SARS-CoV-2 RNA detection in pooled saliva samples: An effective cost-saving method

Jiraphat Charoenkupt — 2024-10-01

Background: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a causative agent of coronavirus disease 2019 (COVID-19), which was first reported in December 2019 and has since spread globally. Effective laboratory testing is crucial for the early detection and prevention of SARS-CoV-2 trans mission. The use of pooled saliva samples represents a potential method for active case finding and increasing testing efficiency.

Objectives: To evaluate the effectiveness of pooled saliva samples for SARS-CoV-2 testing and its costeffectiveness for screening healthcare workers at King Chulalongkorn Memorial Hospital, Thai Red Cross Society.

Methods: A total of 24,098 samples collected between April 19, 2021, and May 30, 2022, to be tested for the presence of SARS-CoV-2 were analyzed retrospectively. The samples were examined individually and in pools of four and six using the Cobas 6,800 reverse-transcription polymerase chain reaction assay for the detection of SARS-CoV-2 RNAs. The analysis focused on changes in cycle threshold values for each target between positive pools and positive individual samples.

Results: SARS-CoV-2 was detected in 0.5% of the samples (123/24,098). Pooling saliva samples in groups of four or six did not compromise the detection of viral RNAs. Pooled saliva testing showed high performance for SARS-CoV-2 detection, with cost reductions of 73.5% for the four-sample pools and 80.7% for the sixsample pools compared with individual testing.

Conclusion: Pooling saliva samples is a cost-effective and efficient method for screening SARS-CoV-2, particularly in low-prevalence settings. This approach helps quickly identify and isolate healthcare workers with infection, thus reducing transmission and preserving resources.

Clinical performance of a multistep algorithm for the diagnosis of Clostridioides difficile infection and patient characteristics in a clinical setting

Mayuree Khantipong — 2024-10-01

Background: Clostridioides difficile infection (CDI) is a significant cause of antibiotic-associated diarrhea and colitis. Its diagnosis relies on clinical presentations confirmed by laboratory investigations.

Objectives: This study aimed to evaluate the performance of current diagnostic tests and examine the characteristics of patients with CDI in a real hospital setting.

Methods: In total, 299 unformed stool specimens were collected and analyzed using the C. DIFF QUIK CHEK COMPLETE and polymerase chain reaction (PCR) assays. Patient data were retrospectively reviewed from medical records.

Results: The C. DIFF QUIK CHEK COMPLETE and PCR assays detected toxigenic C. difficile in 16/299 (5.4%) and 37/299 (12.4%) specimens, respectively. The agreement rates between these two assays for detecting C. difficile and toxin A/B were 90.3% and 93.0%, respectively. The use of a multistep algorithm with the C. DIFF QUIK CHEK COMPLETE assay, arbitrated by PCR assays, significantly increased the detection of toxigenic C. difficile (P < 0.05). Among the clinical characteristics of patients, age > 60 years was significantly associated with CDI (P < 0.05). However, the duration of antibiotic exposure and antibiotic type were not significantly different between patients with and without toxigenic C. difficile. In addition, C. difficile diagnostic tests and treatments are inappropriately used among patients presenting with diarrhea of other causes and a history of antibiotic exposure.

Conclusion: A multistep algorithm is a valuable diagnostic tool for CDI, particularly in hospitals without established testing criteria. To prevent the inappropriate utilization of laboratory resources, effective stewardship of C. difficile testing is essential.

Phytochemical profiling and antioxidant potential of methanolic extract of Vitis vinifera L seeds

Sindhuja Lakshmi — 2024-10-01

Background: Grapes are the most widely consumed fruit worldwide. Grape seeds are a natural source of polyphenols that possess pharmacological and therapeutic activities. The grape seeds have long been used in traditional medicine to alleviate various illnesses.

Objective: This study aimed to identify the major phytocompounds of grape seed methanolic extract and evaluate its antioxidant activity.

Methods: The methanolic extract of grape seeds was subjected to gas chromatography–mass spectrometry to identify the major phytochemical compounds, and further antioxidant activity was evaluated using 2, 2- diphenyl-1-picrylhydrazyl (DPPH) as per the guidelines of the Association of Official Analytical Chemist.

Results: In total, 10 phytochemical compounds (1, 3, 5-benzenetriol, 1, 3-benzenediol, benzoic acid, reticuline, 2-hydroxy-5-methylisophthalaldehyde, 4-vinylbenzene-1, 2-diol, cyclohexane carboxylic acid, eicosanoic acid, octadecanoic acid, and 2, 6, 10, 14, 18-pentamethy l - 2, 6, 10, 14, 18-eicosapentaene) were identified in the methanol extract. By the DPPH method, the antioxidant activity of the extract was 212.6 µg/kg.

Conclusion: Phytochemical profiling of the methanolic extract of grape seeds revealed the presence of phenols (1,3-benzenediol, 4-vinylbenzene-1,2-diol), polyphenol (1,3,5-benzenetriol), phenolic acid (benzoic acid), alkaloid (reticuline), aldehyde (2-hydroxy-5-methylisophthalaldehyde), straight-chain saturated fatty acid (eicosanoic acid and octadecanoic acid), and carboxylic acid (cyclohexane carboxylic acid). Of these phytocompounds, benzoic acid and reticuline are potent natural antioxidants that would be effective against free radicals/reactive oxygen species with wider therapeutic applications in metabolic disorders, inflammatory conditions, and cancer.