Medical Laboratory Technology Journal

Fibrinolytic Protease-Producing Bacteria with Varied Hemolysis Pattern Associated with Marine Algae Dictyota sp.

Muhammad Ardi Afriansyah — 2023-12-23

The main death factor of cardiovascular disease (CVD) is the formation of a blood clot (thrombus). Thrombus is formed by the action of fibrin, playing a role as a blood coagulation agent. Administration of fibrinolytic enzymes can degrade fibrin through the fibrinolysis process. Therefore, searching for new sources of fibrinolytic enzymes becomes critical in eradicating diseases by fibrinolysis of thrombus. This study aims to isolate fibrinolytic protease-producing bacteria associated with fermented brown algae products Dictyota sp, of Awur Bay, Jepara, Indonesia, and to observe their hemolysis pattern. As many as 14 unique bacterial colonies previously isolated from fermented Dictyota sp. were sub-cultured using Zobell Agar (ZA) medium. Skim Milk Agar (SMA) and Fibrin Agar (FA) were then used as selective media to detect the presence of fibrinolytic protease-producing bacteria, which was indicated by their ability to form a clear proteolytic and fibrinolytic zone simultaneously around bacterial colonies. Hemolysis characteristics of fibrinolytic bacteria were determined using Blood Agar Plate (BAP) to test their ability to produce hemolysin toxin. As a result, of these 14 isolates, 3 of them, namely FD-09, FD-13, and FD-14 (FD= Fermented Dictyota), could produce both proteolytic and fibrinolytic zone with a fibrinolytic index range of 2.0–2.9. Isolate FD-09 is the least pathogenic (g-hemolytic) compared to other fibrinolytic isolates, FD-13 (b-hemolytic) and FD-14 (a-hemolytic), in terms of hemolysin toxicity. In conclusion, fermented Dictyota sp. is a potential source of bacteria-producing fibrin-degrading protease with varied hemolysis patterns. It is necessary to identify bacteria-producing fibrinolytic protease isolates Dictyota sp. and further characterization regarding the specificity and activity of the resulting protease to develop its potential as an antithrombotic agent.

Clinico-microbiological and Risk Factor Analysis of Strongyloides stercoralis Infections in South India

Abirami lakshmy Jayachandran — 2023-12-23

Strongyloides stercoralis is a soil-transmitted nematode infection. It is implicated in causing hyper infection syndrome in immunosuppressed patients. This infection can also present without symptoms. The study aims to analyze the demographic data and risk factors associated with acquiring this infection and detectStrongyloides stercoralis in the stool sample by microscopy and modified agar plate culture. Combining microscopy and culture can facilitate detecting the infection in cases of low worm burden. Evaluation of socio-demographic information and risk factors will help in implementing control measures. A total of 986 stool samples were received. All the samples were examined by saline and Iodine mount examination. All the stool samples were cultured by the modified agar plate method. The risk factors, patient demographic details and clinical features were analyzed for significant association using the Chi-square test and a p-value < 0.05 was considered significant) by quick calls graph pad software. The overall prevalence of Strongyloides stercoralis infection was 22(1.23%), and all these cases were diagnosed on stool examination by the demonstration of rhabditiform larva ofStrongyloides stercoralis and adult females by modified agar plate culture. The positivity for Strongyloides larva was high among males, patients from rural areas and individuals working in agricultural settings. There was a significant association between walking barefoot and the demonstration of the larva. Patients presenting from endemic regions, even with vague, nonspecific gastrointestinal symptoms, especially with risk factors like Diabetes mellitus, should be screened for this infection and treated. Awareness, proper hygiene practices, and appropriate treatment are necessary to prevent this soil-transmitted helminthic infection.

Effect of Soybean Extract on sFlt-1 LEVELS in Huvecs Cultures Induced by Preeclampsia Plasma

Wiwit Indriyani — 2023-12-23

Preeclampsia is a dangerous pregnancy issue that causes hypertension at ≥20 weeks of gestation. Oxidative stress is known to play a role in the pathophysiology of preeclampsia by raising the activity of soluble fms-like tyrosine kinase 1 (sFlt-1) and causing endothelial dysfunction. As a result, antioxidants are utilized as a therapy in preeclampsia to protect the body from the impacts of free radicals. This study aims to determine the effect of soybean extract on sFlt-1 levels in HUVECs cultures exposed to preeclamptic plasma. The benefit of this study is the consideration for clinicians to provide soybean ethanol extract supplements (Glycine max) to increase the potential of antihypertensive therapy. Human umbilical vein endothelial cell cultures (HUVECs) were used in the study, which was conducted in vitro. HUVECs cultures were exposed to preeclampsia plasma and subsequently treated for 24 hours with various dosages of soybean extract. The enzyme-linked immunosorbent assay (ELISA) measured the sFlt-1 levels in each observation group. The hypothesis was tested using One-Way ANOVA analysis with SPSS version 25 software. This study found a significant difference (p <0,05) between the mean value of the positive control group and the negative control group. A significant difference was also found (p <0,05) in the mean value of sFlt-1 between the positive control group and the treatment group that was given soybean extract at doses of 17,5 ppm. Soybean extract had a significant effect on decreasing sFlt-1 levels in HUVECs cultures exposed to preeclamptic plasma. Thus, soybeans isoflavones have the potential to treat preeclampsia by reducing anti-angiogenic factors.

Effect of Vitamin C Exposure to Fibroblast Cells on Woman Uterosacral Ligament Culture

Rahajeng Rahajeng — 2023-12-23

Pelvic Organ Prolapse (POP) occurs when the uterosacral ligaments weaken. Vitamin C plays a major role in forming the extracellular matrix (collagen) and stimulates fibroblast cell proliferation. This study aimed to examine how a woman's uterosacral ligament culture was affected by vitamin C exposure to fibroblast cells. A straightforward experimental design investigation that is non-blind and includes analysis after exposure (Post-test control group design). This study's sample used uterosacral ligament from a woman who underwent at least two vaginal deliveries through a hysterectomy procedure. Several doses of vitamin C were given and repeated for 72 hours, followed by examining the number of fibroblast cells; then, the data was analyzed starting with the parametric prerequisite test and the one-way ANOVA test. There was a significant effect of vitamin C exposure on fibroblast cells of the uterosacral ligament at a dose of 50 μg/ml and 100 μg/ml. In contrast, at doses of 150 μg/ml and 200 μg/ml, there was no significant effect. From this study, it is hoped that the administration of vitamin C according to the optimal dose found in the study (significant impact at doses of 50 μg/ml and 100 μg/ml, can increase the proliferation of fibroblast matrix supporting the sacrouterine ligament in women, to prevent prolapse of the pelvic organs in women, and can maintain the quality of life of women.

Effectiveness of Rosmarinus officinalis and Centella asiatica Nanoemulsions Against Caspase 3 Gestational Diabetes Mellitus Expression

Sutini Lamadi — 2023-12-23

Gestational diabetes mellitus (GDM) is closely related to oxidative stress conditions in insulin resistance conditions that increase Reactive Oxygen Species (ROS) against the body's defense antioxidant mechanism. Chronic complications due to hyperglycemia in patients with GDM increase BAX / BCL2 levels which then activates the change of procaspase 3 to caspase 3, an activator of apoptosis. This study aims to determine the effect of the combination of Rosmarinus officinalis and Centella asiatica nanoemulsions on caspase 3 expressions in zebrafish models of gestational diabetes mellitus, with a posttest-only controlled group design. The samples were divided into five groups, namely K- (EM), K+(EM+3% Glucose), P1, P2 and P3 (3% Glucose + Combination of Rosmarinus officinalis and Centella asiatica 2.5μg/ml, 5μg/ml and 10μg/ml), which will be tested for PEPCK and Caspase expression at the age of 3dpf using Real Time-PCR. The results of the correlation test of the group with caspase 3 obtained (p = 0.045) negative direction. These results show that the combination of Rosmarinus officinalis and Centella asiatica nanoemulsion decreased Caspase 3 expression in zebrafish models of gestational diabetes mellitus. The combination of Rosmarinus officinalis and Centella asiatica nanoemulsion has the potential to reduce blood glucose levels and reduce the risk of apoptosis in gestational diabetes mellitus patients.

Identıfıcatıon of KRAS Mutatıons in Colorectal Carcınoma Patıents at Dr. M. Djamıl Hospıtal, West Sumatra-Indonesıa

Rita Maliza — 2023-12-23

Kirsten rat sarcoma viral oncogene (KRAS) gene mutations lead to abnormal activation of the RAS signalling pathway and have been associated with poor prognosis and resistance to some therapeutics. This study aimed to identify mutation characteristics of the KRAS genes codon 12 and 13 in colorectal cancer patients in West Sumatra. KRAS mutations were analyzed in 20 DNA of colorectal cancer patients' tissue samples by using polymerase chain reaction (PCR) with specific primer and direct sequencing analysis. Our findings showed five samples (25%) with mutated KRAS at codons 12 and 13 (including three samples with GGT→GAT, one sample with GGT→GTT in codon 12, and one sample with GGC→GAC in codon 13). In conclusion, we found two variations of amino acid changes at codon 12 (G12D and G12V) and one at codon 13 (G13D). More research with many samples is required to obtain conclusive data on the relationship between these gene mutations and colorectal cancer response to therapy and prognosis.

HLA-DQB1*05:02 Allele Association with Anti-Tuberculosis Drug Induced Liver Injury: A Single-Hospital Based Study in Jakarta, Indonesia

Kinasih Prayuni — 2023-12-23

Past studies have delved into the genetic factors underlying anti-tuberculosis drug-induced liver injury (AT-DILI), primarily concentrating on polymorphisms in genes responsible for drug-metabolizing enzymes. However, the immune system's potential impact on drug adverse effects, specifically through genes such as HLA, has received limited attention. Previous research has notably revealed an association between HLA-DQB1*05 and AT-DILI, specifically the prevalence of the HLA-DQB*05:02 allele in AT-DILI patients. In light of this, our study aimed to investigate a potential link between HLA-DQB1*05:02 alleles and AT-DILI. In this study, we included 51 AT-DILI cases and 59 controls belonging to the Javanese ethnic group. The HLA-DQB1*05:02 genotypes were determined using a customized PCR-based typing method, and the results were further confirmed by analyzing five samples via the Luminex assay. Our findings revealed a significant association between HLA-DQA1*05: 02 and the risk of AT-DILI (P = 0.022; OR (95% CI) = 6.11 (1.25-29.74)). Moreover, the consistent results obtained from the Luminex assay validated the reliability of the custom PCR-based genotyping approach. This preliminary study sheds light on the relationship between the HLA-DQB1*05:02 allele and AT-DILI within the Indonesian population. Furthermore, our study demonstrates the dependability of custom PCR-based genotyping in detecting HLA-DQB1*05:02 alleles. Nevertheless, further research is imperative to corroborate and expand upon our findings.

Impact of Soybean Phytoestrogen-Rich Extract on Markers of Inflammation Markers in 4-Vinyl Cyclohexane Diepoxide-Induced Menopause in Albino Rats

Edusola Juliana Olaniyan — 2023-12-23

The vasomotor symptoms of menopause, including hot flashes, sweating, physical and psychological discomfort, and emotional changes, are accurate and experienced by many of the menopausal and postmenopausal female population. In addition, it causes osteoporosis and slowed metabolism, raising the chance of developing many different ailments. Given that hormone replacement therapy (HRT) has been linked to an increased cancer risk, this investigation was undertaken to identify viable alternatives. The study aimed to investigate the impact of Soybean phytoestrogen-rich extract on some markers of inflammation of 4-vinyl cyclohexene diepoxide-induced menopause in albino rats. Sixty-five (65) female albino Wistar rats were employed in the investigation. Each one was induced with 80mg/kg of 4-vinyl cyclohexene diepoxide before being treated with either normal estradiol therapy (14ug/kg) or varying concentrations of the soybean phytoestrogen-rich extract (200 mg/kg, 400 mg/kg, and 600 mg/kg). Inflammatory markers (C-reactive protein (CRP), interleukin-6 (IL-6), and tumour necrosis factor-alpha (TNF-α)) were measured by ELISA methods. Statistical software SPSS (IBM) version 23.0 was used to analyze the data. Compared to the positive control group, the soybean phytoestrogen-rich extract therapy group saw a dose-dependent reduction in CRP and IL-6 levels (p<0.05) but not in TNF-α (P>0.05). Data from this research demonstrate the anti-inflammatory effects of soybean phytoestrogen-rich extract therapy in menopause-induced female Wistar rats. Soybean phytoestrogen-rich extract therapy in a high dose appears to show no significant difference from hormone replacement therapy as an alternate estrogen source in managing inflammation as a chronic disease.

Potential Of Mahkota Dewa Fruit (Phaleria macrocarpa) Flavonoids Extract and Dienogest to Reduce IL-17A Levels in Mice Endometriosis Model

Ana Paramita Prastiwi — 2023-12-23

Endometriosis is an inflammatory disease that occurs in glands outside the endometrium. Interleukin 17 (IL-17) is a proinflammatory cytokine degrading Th17 cells, which play an essential role in several inflammatory diseases. IL-17 expression is upregulated in serum, peritoneal fluid (PF), and endometriotic lesions from patients with endometriosis. Usually, treatment for endometriosis is dienogest. However, it has many side effects if used long-term. Fruit and seeds of Phaleria macrocarpa have various critical biological activities in the form of extracts such as antimicrobial, anti-inflammatory, and antioxidant activities. This study aims to analyze the effect of comparing flavonoid fruit extract of Phaleria macrocarpa and dienogest on IL-17A levels in endometriosis mice. A total of 35 endometriosis model mice were divided into seven groups, namely the group of healthy mice (without any treatment), the endometriosis group, and the endometriosis group, which were given the treatment of flavonoid fruit extracts of Phaleria macrocarpa and dienogest. The sample used in this study was peritoneal fluid. The enzyme-linked immunosorbent assay (ELISA) measured IL-17A levels in each observation group. The hypothesis was tested using One-Way ANOVA analysis with SPSS software version 26. This study found a significant difference (p<0.05) between the mean values of the positive control group and the negative control group. There was also a significant difference (p<0.05) in the mean value IL-17 between the treatment groups that were given sphalerite macrocarpa and dienogest extracts. Flavonoid fruit extract of Phaleria macrocarpa had a significant effect on reducing IL-17A levels in endometriosis model mice, and there is a significant difference with dienogest. Thus, the flavonoid fruit extract of Phaleria macrocarpa can potentially treat endometriosis by decreasing inflammatory factors.

Bioactivity Examination of Uncaria gambir (W.Hunter) Roxb on In Vitro Human Sperm Motility

Silvani Permatasari — 2023-12-23

Globally, 48 million couples experience infertility, where male infertility factors contribute to 50% of cases. Spermatozoa motility is a crucial parameter in assessing male fertility. Antioxidants act as the body's defence against excessive ROS and can be used as a treatment for male infertility. One of the local plants in Central Kalimantan that is potentially rich in antioxidants is Bajakah Kalalawit (Uncaria gambir (W.Hunter) Roxb. However, there is limited research on the bioactivity of Bajakah Kalalawit on spermatozoa in vitro. This study aims to determine the effect of ethanol extract of Uncaria gambir (W. Hunter) Roxb on spermatozoa motility in vitro. Bajakah Kalalawit stems were extracted using a 3x24-hour maceration technique with 96% ethanol. After obtaining the concentrated extract, the secondary metabolite compound content was analyzed through a phytochemical screening. The sample used was in vitro human spermatozoa that were washed and added to Bigger Whitten Whittingham medium, then incubated at 37°C for 1 hour with a 96% ethanol extract of Bajakah Kalalawit at doses of 50ng/ml, 100ng/ml, 500ng/ml, and 1000ng/ml, as well as a control group (Bigger, Whitten & Whittingham medium only). The results showed that the compounds contained in the ethanol extract of Uncaria gambir (W.Hunter) Roxb were terpenoids, flavonoids, phenolics, steroids, saponins, alkaloids, and tannins. Spermatozoa motility significantly increased in the treatment groups starting from doses of 50ng/ml, 100ng/ml, 500ng/ml, and 1000ng/ml compared to the control group. Ethanol extract of Uncaria gambir (W.Hunter) Roxb could increase spermatozoa motility in vitro and succeeded in improving reproductive technology.