DOI: 10.3897/folmed.67.e168906

Authors: Muhammad Farid, Hanin Fitri Aqilanisa

Abstract: Abstract Introduction: Varicocele, characterized by the enlargement of scrotal veins, is a common contributor to male infertility, but its genetic underpinnings remain largely unknown. Aim: The goal of this study is to identify potential biomarkers associated with varicocele in order to better understand its molecular mechanisms. Materials and methods: Using the three primary databases, NCBI, DisGeNET, and OpenTarget, we analyzed gene variants and found 79 pertinent genes associated with varicocele. Protein-protein interaction analysis was performed using STRING and visualized with Cytoscape. Molecular Complex Detection (MCODE) and CytoHubba tools helped identify significant protein clusters. Results: The gene ontology analysis shows that there are 79 proteins involved in the inflammatory process, the regulation of gene expression, and cellular components that play a role in oxidative stress and angiogenesis. Our results revealed three key biomarkers: Interleukin-1 beta (IL1B), B-cell lymphoma 2 (BCL2), and matrix metalloproteinase-9 (MMP-9). These proteins are involved in critical processes, such as inflammation, oxidative stress, angiogenesis, and vascular damage, that are central to the pathophysiology of varicocele. Conclusion: The identification of IL1B, BCL2, and MMP-9 offers new insights into varicocele’s molecular mechanisms and suggests potential targets for diagnostic and therapeutic strategies, advancing personalized treatment approaches for fertility restoration.

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DOI: 10.3897/folmed.67.e153542

Authors: Machineni Sravani, Manickam Kokila, Kasinathan Ramanathan, Arun Kumar

Abstract: Heart failure (HF) poses a major global health burden due to its high prevalence, complexity, and poor prognosis. Although biomarkers such as B-type natriuretic peptides (BNP, NT-proBNP) are widely used for diagnosis and risk stratification, additional biomarkers are needed to refine prognostication. Copeptin, a stable fragment of pre-provasopressin, reflects vasopressin system activity and has emerged as a promising prognostic tool. Elevated copeptin levels correlate with increased mortality, hospitalizations, and disease progression in both acute and chronic HF. It offers early detection of hemodynamic stress and complements traditional markers, especially in multimarker strategies. This review explores copeptin’s physiological role, its predictive value in various HF phenotypes, and its integration into clinical risk models. Evidence supports its utility in identifying high-risk patients, guiding therapy, and monitoring disease evolution. Challenges to clinical adoption include assay standardization, cost-effectiveness, and establishing universally accepted cutoffs. Future directions focus on copeptin-guided therapies, AI-driven predictive models, and its role in precision medicine. Continued research may solidify copeptin’s role in optimizing heart failure management through individualized risk assessment and tailored interventions.

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DOI: 10.3897/folmed.66.e117367

Authors: Amba Esakki, Anitha Pandi, Smiline A S Girija, Vijayashree Priyadharsini Jayaseelan

Abstract: Introduction: Transformer (TRA2B) is a serine/arginine-rich (SR)-like protein family that regulates the alternative splicing of several genes in a concentration-dependent manner. Amplification of the TRA2B gene, which codes for TRA2B, occurs in several malignancies, including those of the lung, cervix, head and neck, ovary, stomach, and uterine. Materials and methods: The present study design follows a computational approach to predict the molecular mechanisms underlying TRA2B alterations in two cancer phenotypes, viz., lung and head and neck squamous cell carcinoma. The genetic alteration in the TRA2B gene was identified using the cBioportal database. The gene expression pattern in both the cancer types and their survival pattern concerning the expression profile was demonstrated using UALCAN. The microRNA targets of the TRA2B gene were identified using the miRDB database. Results: Genetic alteration was found to be 27% and 48% in HNSCC and LUSC datasets, respectively. The alterations included gene amplification, missense, nonsense, and splice site mutations. The gene expression profile of TRA2B correlated well with the gene amplification demonstrated by patients in both groups. However, the upregulation of TRA2B did not correlate well with the survival profile in LUSC patients. The downregulation of TRA2B markedly affected the survival of HNSCC patients, which can be attributed to the functions of microRNAs targeting TRA2B transcripts. Conclusion: Although TRA2B was found to be a potential diagnostic marker exhibiting a differential expression pattern for HNSCC, the employability of this gene as a prognostic marker needs more experimentation. Also, the influence of microRNAs on dysregulated gene expression should be considered to gain a better understanding of the underlying molecular mechanisms precipitating the disease.

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DOI: 10.3897/folmed.66.e124561

Authors: Sahar Mohammadi Baladezaee, Mehrdad Gholami, Elham Amiri, Hamid Reza Goli

Abstract: Aims: Pseudomonas aeruginosa plays an important role in hospital infections caused by several virulence factors, such as elastase and proteases. This study aimed to evaluate the prevalence of LasA, LasB, and PIV genes, encoding these enzymes, in clinical isolates of P. aeruginosa. Materials and methods: One hundred clinical isolates were collected from patients admitted to educational and therapeutic hospitals of Mazandaran Province, North Iran. The isolates were identified by the standard microbiological and biochemical tests. The bacterial DNA was extracted by the alkaline lysis method, and the presence of relevant genes was detected using the PCR method. The data were analyzed using SPSS v. 23 and the chi-square test. A p-value <0.05 was considered statistically significant. Results: In 100 clinical isolates of P. aeruginosa, the LasA, LasB, and PIV genes were presented with a frequency of 97%, 96%, and 97%, respectively. Of the total number of samples, 39 patients were female and 61 were male. Also, the majority of the patients were between 61 and 70 years old. Conclusion: LasA, LasB, and PIV are highly prevalent in clinical isolates of P. aeruginosa, indicating the importance of these genes as key virulence factors in P. aeruginosa pathogenicity in this region.

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DOI: 10.3897/folmed.66.e113356

Authors: Steliyan Petrov, Hristo Taskov, Marianna Murdjeva

Abstract: The COVID-19 pandemic has left a lasting impact on global health, challenging communities, healthcare systems, and researchers worldwide. As we navigate this unprecedented crisis, this paper embarks on a multifaceted exploration of the pivotal role played by natural killer (NK) cells in the context of COVID-19. A significant portion of this paper is devoted to dissecting the nuanced role that NK cells assume in the context of COVID-19. From the initial acute infection to post-recovery immunity, NK cells emerge as critical players. We scrutinize the activation and dysregulation of NK cells during SARS-CoV-2 infection, shedding light on their potential contribution to disease severity. Moreover, we explore the fascinating landscape of post-COVID immunity, where NK cells are known to interact with adaptive immune responses, providing a foundation for long-term protection. In light of their central role, we investigate therapeutic strategies targeting NK cells in COVID-19 management, presenting an overview of current research efforts and their promise in mitigating disease progression. Lastly, we draw attention to research gaps, emphasizing the need for further investigation into NK cell dynamics during COVID-19. These gaps represent opportunities for advancing our understanding of NK cell biology and, by extension, enhancing our strategies for combating this global health crisis. This comprehensive exploration not only highlights the intricate interplay between NK cells and the COVID-19 pandemic but also underscores the importance of these innate immune warriors in shaping both the acute response and long-term immunity, ultimately contributing to the broader discourse surrounding the pandemic’s pathophysiology and therapeutic approaches.

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DOI: 10.3897/folmed.65.e96012

Authors: Oliver Oey, Siaw Sze Tiong, Sze Ling Wong, Suresh Navadgi, Yasir Khan

Abstract: Cancers that develop within six months of the first primary cancer are referred to as synchronous malignancies. These malignancies are difficult to diagnose and treat, with treatment primarily based on case reports. We report here the case of a 51-year-old male with prior history of prostate cancer who presented with haematuria to the general practice. A CT pyelogram showed left bladder wall lesion that was further investigated with cystoscopy and biopsy confirmed as muscle-invasive urothelial carcinoma. Incidentally, two perigastric nodes and hepatic lesions were noted on CT. FDG-PET revealed high-grade uptake in the right lobe of thyroid gland and cervical nodes that was biopsy proven as papillary thyroid carcinoma. Subsequently, gastroscopy and a biopsy of the gastric lesion confirmed a gastric neuroendocrine tumour. The patient underwent chemoradiotherapy, total thyroidectomy, and commenced somatostatin analogue for treatment of urothelial carcinoma, papillary thyroid carcinoma, and neuroendocrine tumour, respectively. The diagnosis and treatment of synchronous malignancies is complex. A multidisciplinary team approach is required to improve treatment outcomes.

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DOI: 10.3897/folmed.65.e81316

Authors: Ilter Demirhan, Erkan Oner, Ergul Belge Kurutas

Abstract: Introduction: The rising rate of childhood obesity and the serious health problems it causes are gaining increasing attention in medical research and health policy. Aim: This study aimed to evaluate the relationship between insulin resistance and the oxidative stress biomarker 8-iso-prostaglandin F2α levels in obese children. Materials and methods: Forty-four children in total (21 boys and 23 girls) aged between 6 and 15 years and diagnosed with obesity who attended the Pediatric Endocrinology Unit between December 2020 and June 21 were enrolled in our study. Forty children (20 boys and 20 girls) without systemic diseases were selected as controls. From the percentile curves determined for Turkish children, percentile values of obese children and control group were calculated based on sex and age. In addition, the insulin resistance values (HOMA-IR) in the homeostasis model were calculated. The relationship between the variables was examined with the Pearson and Spearman correlation tests. Children between the 5th and 85th percentile were defined as the control group, and those above the 95th percentile were defined as the obese group. Systolic and diastolic blood pressure, triglyceride, total cholesterol, LDL cholesterol, HDL cholesterol, fasting blood sugar (glucose), insulin, and 8-iso-PGF2α concentrations were measured in all children included in the study. Results: There were significant differences between the two groups in terms of age, body mass index, and systolic and diastolic blood pressures (p<0.05). Glucose, triglyceride, insulin, 8-iso-PGF2α and HOMA-IR levels were found to be statistically significantly higher in obese children than the levels in the control group (p<0.05). In addition, significant positive correlations were found between insulin levels and glucose, triglyceride and HOMA-IR values in obese patients (p<0.05). In obese children, 8-iso-PGF2α concentrations were found to be statistically significantly higher than those in the control group (p<0.01). ROC analysis had a good diagnostic value for 8-iso-PGF2α where the area under the curve was 1.0. A direct, positive, statistically significant correlation was found between insulin resistance and the 8-iso-PGF2α values (r=0.420, p=0.037). Conclusions: 8-iso-PGF2α concentrations were found to be higher in obese children than in the control group. It was observed that increased insulin resistance raised 8-iso-PGF2α levels. 8-iso-PGF2α is thought to be particularly important for the diagnosis and treatment of these patients, with 99% sensitivity and specificity.

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DOI: 10.3897/folmed.64.e66292

Authors: Larysa Sivitskaya, Tatiyana Vaikhanskaya, Nina Danilenko, Aleh Liaudanski, Oleg Davydenko, Nikolai Zhelev

Abstract: Danon disease (DD), a rare X-linked genetic illness with a poor prognosis, is caused by a mutation in the lysosome-associated membrane protein 2 gene (LAMP2). Three main clinical features of this pathology are cardiomyopathy, skeletal myopathy, and mental retardation. Most Danon disease mutations create premature stop codons resulting in the decrease or absence of LAMP2 protein. The present case reports the frameshift variant c.190_191delАС in the LAMP2 in the family with sudden cardiac death history and three members with cardiomyopathy. The presenting phenotype in a female proband with c.190_191delАС was isolated dilated cardiomyopathy in her thirties whereas in two males, DD presented as hypertrophic cardiomyopathy and mild skeletal myopathy since childhood. To examine the contribution of X-inactivation to cardiomyopathy onset we estimated the X-inactivation status in the heart tissue of the affected female. We observed the random pattern (66:34) with the proportion of cardiomyocytes expressing healthy LAMP2 allele reduced to 34%. Deletion c.190_191delАС has led to a complete loss of function LAMP2 due to a single copy of this gene in males. In a woman, cardiomyopathy developed because of both the LAMP2 mutation and a decrease in the expression of a healthy allele in the heart. Based on the strong association of truncating LAMP2 mutations with DD and phenotypes in affected members, the variant c.190_191delАС was classified as pathogenic.

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DOI: 10.3897/folmed.64.e63599

Authors: Pantelis Dimaras, Oskan Tasinov, Desislava Ivanova, Yoana Kiselova-Kaneva, Nadezhda Stefanova, Maria Tzaneva, Diana Ivanova

Abstract: Introduction: Improving RNA isolation and cDNA synthesis techniques has emerged due to advancements in the knowledge of molecular basis of most diseases. This in turn increased the need of higher quantity and quality of the extracted genetic material to be used for a variety of diagnostic tests and experiments. Aim: The aim of the study was to compare three modified methods for RNA extraction from formalin-fixed paraffin embedded (FFPE) biopsied tissue and different cDNA synthesis strategies to facilitate study of gene expression. Materials and methods: Compared RNA extraction methods were: lysis buffer, phenol-based extraction, and combination of both with concomitant use of silica-based spin columns. RNA quantity and purity were estimated spectrophotometrically. Different priming strategies for cDNA synthesis were applied: oligo dT, combination of oligo dT and random hexamer, and gene specific primer. Two-step RT-qPCR of ribosomal protein L37A on preamplified and non-preamplified cDNA templates was performed. Results: The combination of lysis buffer with phenol based extraction gave higher RNA yield. By doing cDNA preamplification, the confidence of detection by qPCR was raised, and efficiency was improved. The preamplified template increased the sensitivity of analysis. Conclusions: Together, the combination of approaches improved substantially the reproducibility and validity of quantitative gene expression analyses from FFPE tissues.

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DOI: 10.3897/folmed.64.e63624

Authors: Ivva Philipova, Viktoryia Levterova, Ivan Simeonovski, Todor Kantardjiev

Abstract: Introduction: Mycoplasma genitalium is an established cause of sexually transmitted infections in men and women. Current guidelines recommend azithromycin and moxifloxacin as first- and second-line treatment, respectively. However, azithromycin treatment failure has been increasingly reported. The aim of this study was to determine the efficacy of azithromycin and alternative antibiotic regimens in a prospective cohort of M. genitalium-positive patients, and macrolide resistance mutations associated with azithromycin failure. Materials and methods: Consecutive eligible M. genitalium-positive patients attending the National Center of Infectious and Parasitic Diseases in Sofia, Bulgaria between 1 January 2018 and 31 December 2020 were treated with azithromycin and retested by polymerase chain reaction 21-28 days after completion of the treatment. Cure was defined as M. genitalium-negative result on the test of cure. Cases failing azithromycin were treated with moxifloxacin and retested another 21-28 days after treatment. Pre- and post-treatment samples were assessed for macrolide resistance mutations by conventional DNA sequencing. Results: Of 21 patients treated with azithromycin, 11 (52.4%) were cured. Pre- and post-treatment macrolide resistance mutations were detected in 10 (47.6%) patients, and all of them failed azithromycin. Moxifloxacin was effective in all cases failing azithromycin; and all were M. genitalium-negative at the test of cure after moxifloxacin treatment. Conclusions: In this study a high azithromycin failure rate (47.6%) in an M. genitalium-positive cohort in association with high levels of pretreatment macrolide resistance was reported. Moxifloxacin was highly effective in treating macrolide-resistant infections. These findings necessitate implementation of new diagnostic and therapeutic strategies such as sequential antimicrobial therapy for M. genitalium guided by a macrolide-resistance assay.

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DOI: 10.3897/folmed.63.e56786

Authors: Chaitali Lad, Ishan Panchal, Ashish Patel, Afzal Nagani, Vruti Parikh, Harnisha Patel, Bhargav Bhimani

Abstract: Introduction: Malaria is one of the varieties of fatal diseases caused by a protozoan parasite that is now considered to be the greatest global health challenge. A parasite of Plasmodium species triggers it transmitting the disease to humans by the bites of female Anopheles mosquitoes.Aim: To screen out designed molecules by molecular docking analysis and assess their pharmacokinetic properties using SwissADME. To synthesize the designed compounds. To characterize the synthesized compounds by TLC, melting point, IR spectroscopy, mass spectrometry, 1H NMR, and 13C NMR. To evaluate the synthesized compounds for antimalarial activity.Materials and methods: In silico analysis was performed with SWISSADME, and molecular docking was performed by AutoDock Vina version 4.2. In vitro antimalarial activity study was performed.Results: In-vitro studies of synthesized molecules showed that compounds C2 (IC50 1.23), C6 (IC50 0.48), C10 (IC50 0.79), and C14 (IC50 0.19) possess good antimalarial activity.Conclusions: 7-chloroquinoline-piperazine derivatives exhibited potential antimalarial compounds for pf-DHFR inhibitors.

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DOI: 10.3897/folmed.63.e56566

Authors: Faeze Abbaszadeh, Alka Hasani, Mohammad Ahangarzadeh Rezaee, Javid Sadeghi, Akbar Hasani, Mahin Ahangar Oskouee, Abolfazl Vahhabi

Abstract: Introduction: Acinetobacter baumannii infections are a growing public-health concern. The bacterium’s potentiality to acquire resistance to a number of commonly used antibiotics has turned it into a formidable pathogen.Aims: Molecular characterization of extensive drug resistant (XDR) typing of A. baumannii clinical isolates by polymerase chain reaction.Materials and methods: Thirty XDR A. baumannii were investigated for the presence of genes encoding carbapenemase resistance, biofilm capacity, autoinducer synthase, virulence and surface motility by polymerase chain reaction (PCR). Later, the isolates were typed by plasmid-based replicon (Rep) (PBRT) and trilocus sequence typing.Results: All 30 XDR A. baumannii strains displayed genes related to surface motility, autoinducer synthase, virulence determinant, biofilm related genes except PER, and bap, the frequency of which was 83.3% and 76.6%, respectively. Analysis of rep genes showed highest frequency of rep6 and rep2 genes, with frequency of 75% and 65%, respectively. All XDR A. baumannii strains belonged to SG I (European clone II) group.Conclusions: Our results show the extraordinary plasticity of XDR A. baumannii and suggest that the strains have gained endemicity in our hospital, which could be a great concern in the near future.

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DOI: 10.3897/folmed.63.e55627

Authors: Ergul Belge Kurutaş, Mehmet Emrah Aksan, Petek Curuk, Mehmet Akif Curuk

Abstract: Background: Beta thalassemia is one of the most common autosomal single-gene disorders in the world. The prevalence of the disease is in the “thalassemia belt” which includes the Mediterranean region of Turkey; throughout the country the gene frequency is estimated to be 2.1%, but in certain regions, this figure increases to 10%.Aim: In this first study, we aimed to determine the frequency of β-thalassemia trait and distrubition of mutations in Kahramanmaraş province, which is located in the southern part of Turkey.Materials and Methods: In this study; 5 ml blood samples was taken from 14 thalassemic patients and their relatives who were taking care of Sutcu Imam University Hospital at Kahramanmaraş. Also, we collected blood samples from 245 adults for screening beta thalassemia trait. Haematological data were obtained by cell counter. HbA2 was determined by HPLC. Ten common mutations were screened by ARMS (Amplification Refractory Mutation System) method. These β-thalassemia mutations are -30 (T>A), Fsc8 (-AA), Fsc8/9 (+G), IVS1-1 (G>A), IVS1-5 (G>C), IVS1-6 (T>C), IVS1-110 (G>A ), Cd 39 ( C>T), IVS2-1 (G>A), IVS 2-745 (C>G). A rare mutation; Fsc44 (-C) was charecterized by DNA sequencing.Results: Ten patients were detected as homozygous for IVS1-110 (seven cases), Fsc 44 (two cases) and IVS1-5 (only one case). Rest of the 4 patients were double heterozygous (two: IVS1-110/IVS1-6, one: Fsc8/Fsc8-9, one: IVS2-1/IVS1-5). In 245 adult, five β-thalassemia trait were detected by screening survey. Conclusion: Sixteen alleles were detected as IVS1-110 in 57.1%. It was seen the most common mutation in Kahramanmaraş. Seven different β-thalassemia mutations were found in this study. Each of 10 families have only one thalassemic patient, other two families have double thalassemic patient in total 12 family.

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DOI: 10.3897/folmed.63.e55346

Authors: Silva Garo Kyurkchiyan, Todor Miroslavov Popov, Felitsiya Shakola, Julian Rangachev, Vanyo Ivanov Mitev, Radka Kaneva

Abstract: Introduction: Recently, miRNAs have become popular molecules used as non-invasive biomarkers in cancer diseases.Aim: The aim of the study was to explore the expression of four miRNAs isoforms: miR-31-3p, miR-196a-5p, miR-210-3p and miR-424-5p in plasma and tissue samples from patients with advanced laryngeal squamous cell carcinoma (LSCC) and healthy controls.Materials and methods: Fresh-frozen tumour and normal laryngeal tissue as well as plasma samples were obtained from 22 patients diagnosed with advanced LSCC. The control group included plasma samples from 21 cancer-free volunteers. Total RNA (including miRNAs) extraction, reverse transcription and real time qPCR were the laboratory techniques used in the study. The obtained results were analyzed using SPSS software v. 23.Results: We found that miR-31-3p, miR-196a-5p, and miR-210-3p levels were significantly elevated in laryngeal tumour tissue, but only the levels of miR-31-3p and miR-196a-5p were significantly upregulated in the plasma LSCC target group. Positive correlation was obtained for miR-31-3p (rs=0.443, p=0.039) and miR-196a-5p (rs=0.548; p=0.008) between plasma and adjacent tumour tissue LSCC samples. ROC analyses were used to evaluate the discriminative power of both miRNAs alone and in combination. The combination of miR-31-3p and miR-196a-5p showed best results with AUC=0.978 (95% CI: 0.945–1.000, p<0.001) with 100% sensitivity and 81% specificity at cut-off: RQ=2.99.Conclusions: Based on this miR-31-3p and miR-196a-5p are proposed as potential biomarkers for validation in larger LSCC group and could be included in a non-invasive miRNAs set for detection of advanced LSCC.

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DOI: 10.3897/folmed.63.e52891

Authors: Khushal Kapadiya, Kishor Kavadia, Jyoti Gohel, Ranjan Khunt

Abstract: Introduction: Due to the vast medicinal importance of purine nucleoside, a hybrid molecule of triazole with purine ring might explode a lead molecule in the pharma sector and based on the last decade’s studies suggested that the nitrogen-rich molecules possess a wide range of medicinal importance.Aim: Due to the vast application of purine nucleoside itself in the field of cancer research, we synthesized triazolo[3,4-e]purines and screened them for their anti-cancer study against NCI-60 cell lines by the protocol used by NIH.Materials and methods: The targeted molecules, 4-chloro-5a,6-dihydro-8-substitutedphenyl-1H-[1,2,4]triazolo[3,4-e]purine derivatives (4a-4h) were synthesized in a two-step procedure by nucleophilic substitution (SN) at C-2 chlorine followed by formation of the triazole ring by acid-catalyzed reaction in the polar protic solvent.Results: It was observed that the regioselective approach followed in C-2 chlorine replacement instead of C-6 chlorine during SN reaction. One-dose response of selected three molecules (4a, 4b, and 4c) showed that 4b (K-562: 64.47 µM & SR: 63.38 µM; mean GI50: 99.09 µM) was found to be more potent than 4a and 4c.Conclusions: We have described in this study the general synthetic method for triazolo[3,4-e]purines as an innovative class of potential anticancer agents. The dose-response curve in the sense of mean GI50 for three compounds across all 60 cell lines, 4b can be served as lead after necessary modification.

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DOI: 10.3897/folmed.63.e52655

Authors: Khushal Kapadiya, Piyush Dholaria

Abstract: Introduction: Nowadays, researchers are progressively concentrated to generate economical, affordable and also greener synthesis approach for the synthesis of various heterocycles. On look at the beauty of coumarin molecules and oxazoles, it seems to be lead molecules in the anti-microbial area. Aim: With the target to identify efficient molecules, we studied 2-oxo-2H-chromen-4-yl-2-((5-substituted aryl-1,3,4-oxadiazol-2-yl) thio)acetate derivatives using two synthetic protocol/methods, i.e. conventional synthesis and microwave-based synthesis.Materials and methods: Two simultaneous methods, i.e. conventional and microwave synthesis have been used for the synthe-sis of 2-oxo-2H-chromen-4-yl-2-((5-substituted aryl-1,3,4-oxadiazol-2-yl)thio)acetate (6a-l) derivatives. The desired molecules were synthesized by conventional and microwave synthesis and a comparative study was carried out to identify an easy route for industrial applications. The confirmations of the compounds were carried out by spectroscopic techniques such as IR, 1H NMR, 13C NMR, mass spectra and elemental analysis. Results: All synthesized compounds were evaluated for their in-vitro antibacterial activity against gram-positive bacteria (Staphylococcus aureus, Staphylococcus pyogenes), gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa), and antifungal activity (Candida albicans, Aspergillus niger). Conclusions: All conventional synthesis of final coumarin derivatives were completed within 4-6 h. While that of microwave-based reaction took comparatively more reaction time. Surprisingly, the compounds 6f and 6g could not be synthesized by microwave radiation even after 32 minutes of irradiation. As to the medicinal application part, microbial evaluation of synthesized analogues showed that the compounds 6b, 6e, 6d, and 6j were found more potent in comparison to the reference drug.

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