Indonesian Journal of Medical Laboratory Science and Technology

The effects of varying in incubation time and temperature of methyl salicylate as a clearing agent on the quality of breast tissue slides

2024-11-13T01:42:11+00:00

Non-polar clearing agents have the ability to dissolve breast adipose tissue. While fat dissolution occurs during the clearing process, careful monitoring of adipose cell membranes mis crucial due to their impact on the behavior of breast cancer cells and disease progression. Xylol is a widely employed clearing agent. However, its toxicity and prolonged clearing time necessitate the exploration of alternatives. In this context, methyl salicylate emerges as a viable substitute. It is non-polar nature facilitates efficient fat dissolution, leading to quicker tissues clarification. This study adopted a quasi-experimental method, utilizing 24 pieces of breast tissue as the sample. These tissue sample were divided into six groups according to the treatment regimen. The treatment groups involved breast tissue clearing using xylol for 2x60 minutes at room temperature, methyl salicylate for 2x30 minutes at room temperature, and methyl salicylate at 60°C for 20 minutes, 30 minutes, 2x20 minutes, and 2x30 minutes. The quality of breast tissue slides was assessed by analyzing the color contrast between the nucleus and cytoplasm using ImageJ software, along with the clarity of adipose cell cell membrane using microscopy. The results indicated that prolonged exposure at high temperature resulted in poor quality breast tissue slides. Conversely, tissue clearing with methyl salicylate for 2x30 minutes at room temperature exhibited excellent contrast between the nucleus and cytoplasm, as well as clear adipose cell membranes. Further study is warranted to explore the applicability of methyl salicylate as a clearing agent in tissues with lower fat content.

Ethanol crude leaf extract of weeping fig (Ficus benjamina) causes mild hepatotoxicity in Sprague dawley rats

2024-11-13T01:42:07+00:00

Weeping fig (Ficus benjaminaeaf extract has diverse medicinal properties but little is reported about its hepatotoxicity. This study determined the mean lethal dose (LD₅₀) and investigated the effects of F. benjamina ethanol crude leaf extract on biochemical parameters and liver histology of Sprague Dawley rats. Twenty-nineemale rats weighing 133-204 g were used. The LD₅₀ was determined with nine rats based on Lorke’s method. The experimental groups consisted of twenty rats, divided into four groups of five. Each group received treatment as follows: Control (feed and water only) and low, medium, and high doses (500, 1000, 1500 mg/kg respectively) of the extract orally for 21 days. All animals were weighed and sacrificed using Ketamine intra-peritoneal injection. Blood samples were collected for biochemical parameters of total bilirubin, conjugated bilirubin, aspartate aminotransferase (AST) alanine aminotransferase (ALT), and alkaline phosphatase (ALP). Liver tissues were removed and processed by the formalin-fixed-paraffin wax-embedding method. The tissue blocks were sectioned and subjected to Hematoxylin/Eosin and Masson trichrome staining. The extract’s LD₅₀ was >5000 mg/kg. The rats’ body weights did not change statistically (p=0.985). Total bilirubin (p=0.003), conjugated bilirubin, AST, ALT, and ALP values (p=0.001) increased significantly. The AST high-doseroup had the highest fold increase (4.8). The liver histology showed mild sinusoidal dilation at 500 mg/kg. There was marked hemorrhage and fibrosis at medium and high doses. Although the extract had relatively low acute toxicity, 1000 and 1500 mg/kg doses were associated with mild hepatotoxicity characterized by veno-occlusion disease. The 500 mg/kg dose is safer for medicinal purposes.

Expression of gonadotropin-releasing hormone receptor type-II correlates with proliferation activity in tissue microarray of rare ovarian tumor

2024-11-13T01:42:04+00:00

Activation of Gonadotropin-Releasing Hormone type II Receptors (GnRHR-II) exhibits antiproliferative activity. GnRHR-II is not only expressed exclusively in the pituitary, but also in a variety of tumors. To date, the clinical relevance of GnRHR-II in ovarian tumors is unclear. In addition, there is a lack of literature addressing GnRHR-II in ovarian tumors, especially rare types. This study was conducted to investigare the correlation between GnRHR-II expression with clinicopathology and proliferative activity of rare ovarian tumors. The purpose of this study was an analytic observational study with a cross-sectional design that utilized 18 ovarian tumor samples on tissue microarray (TMA). The expression of GnRHR-II and Ki67 was assessed using immunohistochemical staining (IHC) and observed using the IHC profiler plugin ImageJ software to obtain their respective H-scores. The data was analyzed using independent t-test, ANOVA, Pearson's test, and Fisher's exact test based on data types. The value of p<0.05 was considered statistically significant. GnRHR-II is expressed in various forms in ovarian tumors, including extrapituitary expression. GnRHR-II expression was highest in the sex cord stromal tumor (SCST) group, 110.30 ± 23.89 (p<0.0001). In addition, there was also a significant difference between GnRHR-II expression with age (p<0.001) and the primary tumor (p<0.05), but not with tumor type (p=0.101). There is a correlation between GnRHR-II expression and proliferative activity (r=-0.043, p=0.866). Elevated GnRHR-II expression is significantly correlation with SCST, individuals over 40 years of age, and tumors confined to the ovary and it is correlates with lower proliferative activity, although this correlation is very weak.

Immunomodulatory activity of kersen leaf extract (Muntingia calabura) on diabetic rats: analysis of immune response

2024-11-13T01:41:59+00:00

The immune response to high blood glucose levels leads to an inflammatory response and also produces inflammation mediators. Immunomodulatory functions of Kersen (Muntingia calabura) need further enhancement to ensure that its benefits are more widely recognized by the public. This study aims to determine the immunomodulatory activity of Kersen leaf in inducing and modulating the immune response in diabetic rats. This study was an experimental laboratory with a pre-and post-test with a control group design. The subjects were 30 white rats (Rattus Novergicus Wistar Strain), were treated with extract M. calabura dose 1 (100 mg/kg bw/day), dose 2 (200 mg/kg bw/day), dose 3 (300 mg/kg bw/day). For clinical evaluation, three control groups were formed, including a Normal Control Group, a Diabetes Mellitus (DM) Positive Group, and a DM Positive Group treated with Anti-Diabetic Drugs. The highest amount of IFN-γ concentrations were found in the DM positive control group + antidiabetic drugs (710.3 ± 27.2 ng/mL). The highest number of Nitrit Oxide (NO) concentration was found in the DM positive control group (103.7 ± 10.2 µmol/L). The highest average amount of pancreatic β cell regeneration was found in the normal control group. The DM positive control group and the treatment group had a significant difference (p < 0.05) It means that there is a significant difference in the data of all treatment groups, or these three groups have anti-diabetic activity by repairing or preventing damage to the pancreas organ in DM rats. This study revealed that M. calabura possesses immunomodulatory activity, capable of inducing and modulating immune responses in diabetic rats.

Molecular approach to the characterization of lipase encoding genes from Moraxella sp. SBE01

2024-11-13T01:41:56+00:00

Lipase from Moraxella sp. SBE01 is an expression of the gene encoding lipase. Detection and characterization of the Moraxella sp. SBE01 lipase coding gene is necessary for large-scale lipase production through genetic engineering. This study aimed to observe the molecular weight, amino acid sequence, length, and conserved amino acids in the DNA encoding the lipase gene, with the goal of identifying and characterizing the lipase-coding gene from Moraxella sp. SBE01. The primer design process was conducted to amplify the lipase gene from Moraxella sp. SBE01 using specialized software for sequence alignment and phylogenetic analysis. Amplification was carried out using PCR with the designed primer, forward primer (GTC ATG ATG TAC TTC CAY GGN GGN GG), reverse primer (GGT TGC CGC CGG CDS WRT CNC C). PCR was carried out under pre-denatured conditions at 95°C (3 minutes), followed by 30 cycles of denaturation at 95°C, annealing at 66°C (30 seconds), 70°C elongations (1 minute) and final elongation of 70°C (10 minutes). The PCR results were electrophoresed using 1% agarose gel with a 1 kb DNA marker. The PCR results were sequenced and analyzed for gene and amino acid sequences and the type of lipase expressed. Sequencing resulted in 387 bp of the nucleotide sequence. The gene and amino acid sequences from Moraxella sp. SBE01 had high homology with the gene and amino acid sequences from Moraxella sp. strain TA144. The lipase gene encodes a protein consisting of 129 amino acids and contains a conserved HGG (His-Gly-Gly) motif, which is characteristic of lipases in family IV, also known as the hormone-sensitive lipase (HSL) family. This conserved sequence suggests that the lipase shares structural and functional similarities with other enzymes in the HSL family, playing a key role in lipid metabolism.

Blumea balsamifera and Sargassum aquifolium extracts reduce fatty liver damage through lipid metabolism signalling pathways

2024-11-13T01:41:52+00:00

Non-alcoholic fatty liver disease (NAFLD) is a condition marked by excessive fat accumulation in the liver and poses a significant health challenge. The leaves of Blumea balsamifera and Sargassum aquifolium have been reported to have anti-atherogenic effects. This study aims to determine the effectiveness of B. balsamifera extract (BBLE) and S. aquifolium extract (SAE) in preventing and treating liver fat accumulation in Wistar rats induced by a high-cholesterol diet through the expression of the AMP-activated protein kinase (AMPK)/ Sirtuin 1 (SIRT1)/peroxisome proliferator-activated receptor γ (PPARγ) pathway, and the leptin receptor. The experimental design of this study is laboratory-based, involving, 20 Wistar rats were fed a high-cholesterol diet over a period of 21 days. The rats were divided into four groups for the evaluation of BBLE and SAE effect: negative control (P0): induced with a high-cholesterol diet + distilled water, positive control (P1): induced with a high-cholesterol diet + simvastatin, P2: induced with a high-cholesterol diet + 4 mg/kg/bw BBLE, and P3: induced with a high-cholesterol diet + 4 mg/kg/bw BBLE and 4 mg/kg/bw SAE. The treatment duration extended over three months. Immunohistochemical analyses were performed on liver tissues to measure AMPK, SIRT1, PPARγ, and leptin receptor expression. The results indicated that leptin expression was lower in the BBLE+SAE group compared to the simvastatin group, and differences were significant between the BBLE and BBLE+SAE groups. No significant differences were noted in AMPK, SIRT1, and PPARγ expression between the simvastatin and BBLE+SAE groups (p≥0.05). In conclusion, BBLE and SAE effectively reduce liver lipid accumulation and enhance fat metabolism in hypercholesterolemic rats.

Prevalence and comorbid for late-stage chronic kidney disease (CKD) in patients undergoing continuous ambulatory peritoneal dialysis (CAPD) due to urinary obstruction

2024-11-13T01:41:48+00:00

Chronic Kidney Disease (CKD) is a condition of gradual or chronic decline in kidney function, which is quite severe and caused by various kidney diseases, including urinary obstruction. This disease is progressive and generally irreversible. CKD requires kidney replacement therapy, one of which is continuous ambulatory peritoneal dialysis (CAPD). To determine the prevalence and risk factors for End Stage Renal Disease (ESRD) in patients undergoing CAPD due to urinary obstruction. We performed a retrospective cohort with a cross-sectional study was conducted using secondary data from medical record data of ESRD patients with CAPD accompanied by urinary obstruction at Dr. Saiful Anwar General Hospital, Malang, Indonesia. The prevalence of CKD in patients with CAPD accompanied by urinary obstruction was 6,50% and dominated by males (57,8%) with an age range of 41-50 years (26%). The majority of comorbidities are severely high the Body Mass Index (BMI) (89,0%) and hypertension (80,8%). The location of obstruction is mostly unilateral (5,64%) with mild levels (4,06%). Urinary obstruction is a frequent clinical finding in CKD patients with CAPD. The most common risk factor in this study was hypertension. The prevalence and comorbidities among CAPD patients with Urinary obstruction (UO) are better understood because to this study. It is necessary to recognise its limitations, particularly the small sample size and single-centre design. Future studies should involve more centres and larger patient groups in order to provide a more thorough knowledge of the mechanisms behind the high survival rates among CAPD patients.

Analysis of bone mineral profile and TSH in early non-dialysis stages of chronic kidney disease - a retrospective cross-sectional study

2024-11-13T01:41:45+00:00

The objective of this study is to analyse the levels of Thyroid stimulating hormone (TSH) and the Bone Mineral Profile (Vitamin D, Parathyroid hormone (PTH), calcium, phosphorus, magnesium, and alkaline phosphatase [ALP]) levels in the early stages of chronic kidney disease (CKD) based on Glomerular filtration rate (GFR). A retrospective analysis was conducted involving 247 CKD patients admitted to the nephrology department at Sri Ramachandra Medical College Hospital from January to June 2022. The estimated GFR (eGFR) was calculated utilizing the CKD-EPI formula provided by the National Kidney Foundation. All biomarkers were analysed using automated platforms. The baseline ages for the three groups were 52.5, 68, and 66.5 years respectively (p<0.001). The comparative analysis revealed statistically significant differences solely among Vitamin D, creatinine, PTH, and phosphorus across the three groups. Further correlation analysis demonstrated changes in bore significant correlations with only creatinine, vitamin D, and PTH. This study concludes that in the early stages of CKD, vitamin D followed by PTH appears to be the earliest biomarker for assessing CKD-Mineral and Bone Disorder (CKD-MBD) occurring prior to any alterations in calcium and phosphate levels. As such, early consideration of supplementation may prove beneficial in mitigating disease progression and preventing cardiovascular complications.