An in vivo and in silico evaluation of the hepatoprotective potential of Gynura procumbens: A promising agent for combating hepatotoxicity.

INTRODUCTION: The liver, the most important metabolic organ of the body, performs a wide variety of vital functions. Hepatic cell injury occurs by the activation of reactive oxygen species (ROS) that are generated by carbon tetrachloride (CCl4), xenobiotics, and other toxic substances through cytochrome P450-dependent steps resulting from the covalent bond formation with lipoproteins and nucleic acids. Observing the urgent state of hepatotoxic patients worldwide, different medicinal plants and their properties can be explored to combat such free radical damage to the liver. In vivo and in silico studies were designed and conducted to evaluate the antioxidant and hepatoprotective properties of Gynura procumbens in rats. MATERIALS AND METHODS: Gynura procumbens leaves were collected and extracted using 70% ethanol. The required chemicals CCl4, standard drug (silymarin), and blood serum analysis kits were stocked. The in vivo tests were performed in 140 healthy Wister albino rats of either sex under well-controlled parameters divided into 14 groups, strictly maintaining Institutional Animal Ethics Committee (IEAC) protocols. For the histopathology study, 10% buffered neutral formalin was used for organ preservation. Later the specimens were studied under a fluorescence microscope. In silico molecular docking and absorption, distribution, metabolism, excretion, and toxicity (ADMET) studies were performed, and the results were analyzed statistically. RESULTS AND DISCUSSION: Gynura procumbens partially negate the deleterious effect of carbon tetrachloride on normal weight gain in rats. The elevated level of serum glutamate pyruvate transaminase (SGPT), serum glutamate oxaloacetate transaminase (SGOT), alkaline phosphatase (ALP), creatinine, LDH, total cholesterol (TC), low-density lipoprotein (LDL), triglycerides (TG), malondialdehyde (MDA), deoxyribonucleic acid (DNA) fragmentation ranges, gamma-glutamyl transferase (γ-GT) in CCl4 treated groups were decreased by both standard drug silymarin and G. procumbens leaf extract. We have found significant & highly significant changes statistically for different doses, here p<0.05 & p<0.01, respectively. On the other hand, G. procumbens and silymarin displayed Statistically significant (p<0.05) and high significant(p<0.01) increased levels of HDL, CAT SOD (here p<0.05 & p<0.01 for different doses) when the treatment groups were compared with the disease control group. Because the therapeutic activity imparted by plants and drugs accelerates the movement of the disturbed pathophysiological state toward the healthy state. In the molecular docking analysis, G. procumbens phytoconstituents performed poorly against transforming growth factor-beta 1 (TGF-ß1) compared to the control drug silymarin. In contrast, 26 phytoconstituents scored better than the control bezafibrate against peroxisome proliferator-activated receptor alpha (PPAR-α). The top scoring compounds for both macromolecules were observed to form stable complexes in the molecular dynamics simulations. Flavonoids and phenolic compounds performed better than other constituents in providing hepatoprotective activity. It can, thus, be inferred that the extract of G. procumbens showed good hepatoprotective properties in rats.

Morphological, Spectroscopic and Thermal Analysis of Cellulose Nanocrystals Extracted from Waste Jute Fiber by Acid Hydrolysis.

Natural cellulose, a sustainable bioresource, is highly abundant in nature. Cellulosic materials, particularly those that explore and employ such materials for industrial use, have recently attracted significant global attention in the field of material science because of the unique properties of cellulose. The hydroxyl groups enable the formation of intra- and inter-molecular hydrogen bonding and the arrangement of cellulose chains in a highly ordered crystalline zone, with the remaining disordered structure referred to as an amorphous region. The crystalline areas of cellulose are well-known as cellulose nanocrystals (CNCs). In the present study, we extracted CNCs from pure cellulose isolated from waste jute fibers by sulfuric acid hydrolysis, followed by characterization. Pure cellulose was isolated from jute fibers by treating with sodium hydroxide (20% w/w) and anthraquinone (0.5%) solution at 170 °C for 2 h, followed by bleaching with chlorine dioxide and hydrogen peroxide solution. CNCs were isolated from pure cellulose by treating with different concentrations (58% to 62%) of sulfuric acid at different time intervals (20 min to 45 min). The FTIR study of the CNCs reveals no peak at 1738 cm-1, which confirms the absence of hemicellulose in the samples. The CNCs obtained after 45 min of acid hydrolysis are rod-shaped, having an average length of 800 ± 100 nm and width of 55 ± 10 nm, with a high crystallinity index (90%). Zeta potential significantly increased due to the attachment of SO42- ions on the surface of CNC from -1.0 mV to about -30 mV, with the increment of the reaction time from 20 min to 45 min, which proved the higher stability of CNC suspension. Crystallinity increased from 80% to 90% when the reaction time was increased from 20 to 45 min, respectively, while a crystallite size from 2.705 to 4.56 nm was obtained with an increment of the acid concentration. Acid hydrolysis enhanced crystallinity but attenuated the temperature corresponding to major decomposition (Tmax) at 260 °C and the beginning of degradation (Ti) at 200 °C due to the attachment of SO42- ions on the surface, which decreased the thermal stability of CNC. The second degradation at 360 °C indicated the stable crystal structure of CNC. The endothermic peak at 255 °C in the DTA study provided evidence of sulfated nanocrystal decomposition and the recrystallization of cellulose I to cellulose II, the most stable structure among the other four celluloses. The proposed easy-to-reproduce method can successfully and efficiently produce CNCs from waste jute fibers in a straightforward way.

A Retrospective Evaluation of Self-Reported Adverse Events Following Immunization with Different COVID-19 Vaccines in Türkiye.

BACKGROUND: The Sinovac and BioNTech vaccines were the first to be introduced in Türkiye to fight the ongoing global COVID-19 pandemic. As these vaccines had shown some side-effects in its clinical trial, we aimed to conduct a survey study to assess the short-term adverse events following immunization (AEFIs) in Türkiye. METHOD: A cross-sectional study was conducted using social and electronic media platforms by delivering a pre-formed and validated online questionnaire among people who had received at least one dose of the COVID-19 vaccine. This survey study focused on mass populations from different regions in Türkiye. A total of 603 responses were collected. Among these, 602 were selected based on complete answers and used for the assessment. The collected data were then analyzed to evaluate the various parameters related to the AEFIs of the respondents. RESULTS: Among the total 602 participants, 20.8% were male, and 78.7% were female, actively answering all of the constructive questions. Most of the respondents were between 18-30 years of age. We found that a total of 23.3% of the total respondents had been infected with the SARS-CoV-2 virus. Our survey revealed that out of 602 volunteers, the rate of experiencing physical discomfort was higher in participants who had received the Pfizer-BioNTech vaccine at all three doses than in those who had received the Sinovac vaccine. When all vaccine types were examined, the most common side effect was pain at the injection site, reported by 75.19% participants. When the side effects were compared according to vaccine types, there was a significant difference only in terms of fever. Fever rates in those who had received the Pfizer-BioNTech vaccine (20.96%) were found to be significantly higher than those who had received the Sinovac vaccine (8%). CONCLUSIONS: The studied vaccines showed minor side effects and there was no significant difference between the vaccines in terms of other side effects. Moreover, further research is needed to determine the efficacy of the existing vaccines in preventing SARS-CoV-2 infections or after-infection hospitalization.

Assessing the Self-reported After Events Following Immunization of COVID-19 Vaccines in Turkey and Bangladesh.

Though mass vaccination programs helped to reduce the severity of the ongoing pandemic, various unwanted effects were reported in Turkey and Bangladesh after taking vaccines. The purpose of this study was to evaluate and compare the adverse effects of several vaccines in Turkey and Bangladesh and how the population of both countries prioritizes the continuation of vaccination compared to the side effects. An online survey with a pretest was conducted to gather data over the research period from July 10, 2021 to December 10, 2021. Finally, the questionnaire was shared with the mass population of Turkey and Bangladesh who have received at least one or two doses of the COVID-19 vaccines. The quality of the questionnaire was evaluated with Cronbach's alpha test. The study consisted of 1508 respondents from Bangladesh and 602 respondents from Turkey. Among the total 2110 respondents, 50.0% were male 66.8% were from the 18-30 years age range, and 77.5% reported living in the city area. Among all the respondents, 64.99% of those vaccinated in Bangladesh and 67.28% of those vaccinated in Turkey reported side effects after vaccinations. Participants receiving mRNA vaccines (Pfizer and Moderna) experienced the most side effects, with many reporting pain at the injection site in both nations. Following that, fever, body pain, and headache were common in Bangladesh, whereas body pain, fatigue, and arm numbness were common in Turkey. The study found no significant adverse events reported in Turkey and Bangladesh following the first and second doses of COVID-19 vaccination. These COVID-19 vaccines showed similar patterns of efficacy and safety during the short period of analysis. Vaccines from different manufacturers showed a non-significant level of adverse events during this binational AEFI approach to COVID-19 vaccines. More studies are recommended on the efficacy and safety of several vaccines to discover unexpected effects.

Exploration of Site-Specific Drug Targeting-A Review on EPR-, Stimuli-, Chemical-, and Receptor-Based Approaches as Potential Drug Targeting Methods in Cancer Treatment.

Cancer has been one of the most dominant causes of mortality globally over the last few decades. In cancer treatment, the selective targeting of tumor cells is indispensable, making it a better replacement for conventional chemotherapies by diminishing their adverse side effects. While designing a drug to be delivered selectively in the target organ, the drug development scientists should focus on various factors such as the type of cancer they are dealing with according to which drug, targeting moieties, and pharmaceutical carriers should be targeted. All published articles have been collected regarding cancer and drug-targeting approaches from well reputed databases including MEDLINE, Embase, Cochrane Library, CENTRAL and ClinicalTrials.gov, Science Direct, PubMed, Scopus, Wiley, and Springer. The articles published between January 2010 and December 2020 were considered. Due to the existence of various mechanisms, it is challenging to choose which one is appropriate for a specific case. Moreover, a combination of more than one approach is often utilized to achieve optimal drug effects. In this review, we have summarized and highlighted central mechanisms of how the targeted drug delivery system works in the specific diseased microenvironment, along with the strategies to make an approach more effective. We have also included some pictorial illustrations to have a precise idea about different types of drug targeting. The core contribution of this work includes providing a cancer drug development scientist with a broad preliminary idea to choose the appropriate approach among the various targeted drug delivery mechanisms. Also, the study will contribute to improving anticancer treatment approaches by providing a pathway for lesser side effects observed in conventional chemotherapeutic techniques.

In Vivo and In Silico Assessment of Diabetes Ameliorating Potentiality and Safety Profile of Gynura procumbens Leaves.

BACKGROUND: Diabetes mellitus is one of the most notable health dilemmas. Analyzing plants for new antidiabetic remedies has become an impressive territory for life science researchers. Gynura procumbens has long been used to treat diabetes. Thus, we strived to ascertain the hypoglycemic potentiality of extract of leaves of G. procumbens by in vivo and in silico approaches. METHODS: Fresh leaves of G. procumbens were collected and shade-dried to prepare ethanolic extracts to evaluate pharmacological parameters. Diabetes was induced in rats via injecting alloxan through the intraperitoneal route at a dose of 150 mg/kg body weight. Humalyzer 3000 was used to perform a biochemical assay of collected samples from rats. Anti-hyperglycemic activity study along with overdose toxicity test was performed. The pharmacological activity of this plant was also evaluated through a molecular docking study. This in silico study investigated the binding affinity of natural ligands from G. procumbens against glycoside hydrolase enzymes. RESULTS: We detected a peak plasma concentration of G. procumbens at 3 hours 45 minutes that is roughly similar to the peak plasma concentration of metformin. Again, in OGTT and anti-hyperglycemic tests, it has been ascertained that both plant extract and metformin can exert significant (P < 0.05) and highly significant (P < 0.01) hypoglycemic activity in a dose-dependent manner. Metformin exhibited better therapeutic efficacy than that of plant extract, but it possessed null statistical significance. Also, our safety profile expressed that, similar to metformin, the plant extract can restore the disturbed pathological state in a dose-oriented approach with a wide safety margin. In silico study also validated the potentialities of natural constituents of G. procumbens. Conclusion. This study suggested that G. procumbens can be considered as potential antidiabetic plant. Robust and meticulous investigation regarding plant chemistry and pharmacology in the future may bring about a new dimension that will aid in discovering antidiabetic drugs from this plant in the diabetes management system.

An evaluation of pharmacological healing potentialities of Terminalia Arjuna against several ailments on experimental rat models with an in-silico approach.

Herbal remedies have been used in many cultures for decades to treat illnesses. These medicinal plants have been found to contain various phytochemical compounds that can help to cure mild to severe illnesses. The inadequacies of conventional medicines and their unusual side effects sparked a determined search for alternative natural therapeutic agents. Another reason for this hunt could be the availability and fewer side effects of natural products. T. arjuna is widely used in traditional medicine to alleviate various diseases like relieving pain, ameliorating diabetes, mitigating inflammation, and back-pedaling of depression. In this study, the ethanolic extract of T. arjuna possesses a promising effect on the animal model (p < 0.05/p < 0.01) as an antihyperglycemic, analgesic, anti-inflammatory, and antidepressant agent, but in a dose-dependent manner. The lower dose of T. arjuna was found to be capable of reversing the disturbed physiological state at a significant level (p < 0.05). However, a higher dose of T. arjuna exerts better therapeutic effects for those diseases. This animal study aims to evaluate the anti-diabetic, anti-depressant, and anti-inflammatory properties of T. arjuna compared to conventional marketed drugs. We will perform an in-silico study for active constituents of T. arjuna against their proposed targets and look for the molecular cascade on their claimed pharmacological properties. This study shows that different doses of T. arjuna bark extracts give similar therapeutic responses compared with established marketed drugs in managing hyperglycemia, stress-induced depression, and inflammation. Besides, our docking study reveals that flavonoids and triterpenoid active constituents of T. arjuna play an important role in its usefulness. This study, therefore, scientifically confirmed the traditional use of this medicinal plant in the management of several diseased conditions.

Current Knowledge Regarding Pharmacological Profile and Chemical Constituents of Gynura procumbens.

Gynura procumbens (Lour.) Merr. is a well-known plant used in folkloric medicine in tropical Asian countries. The plant is prevalently employed by traditional healers in the treatment of diabetes, cancer, hypertension, inflammation, fever, and skin disorders. Several scientific studies reported that Gynura procumbens possesses considerable therapeutic value for the development of emerging treatment options. The diverse pharmacological effects of this plant are attributed to its vast phytoconstituent content. Different chemical classes, including alkaloids, flavonoids, phenolics, steroids, proteins, and polysaccharides, have been isolated from this plant. In this review, we tried to explore the different aspects of Gynura procumbens as an established medicinal plant. The data gathered here give an indication that the plant Gynura procumbens is a good natural source of chemical compounds with different types of pharmacological actions, and these chemical compounds can be used as models for the development of de novo therapeutic agents.

A Retrospective Cross-Sectional Study Assessing Self-Reported Adverse Events following Immunization (AEFI) of the COVID-19 Vaccine in Bangladesh.

BACKGROUND: The Oxford-AstraZeneca vaccine (Covishield) was the first to be introduced in Bangladesh to fight the ongoing global COVID-19 pandemic. As this vaccine had shown some side-effects in its clinical trial, we aimed to conduct a study assessing short-term adverse events following immunization (AEFIs) in Bangladesh. METHOD: A cross-sectional study was conducted on social and electronic media platforms by delivering an online questionnaire among people who had taken at least one dose of the COVID-19 vaccine. The collected data were then analysed to evaluate various parameters related to the AEFIs of the respondents. RESULTS: A total of 626 responses were collected. Of these, 623 were selected based on complete answers and used for the analysis. Most of the respondents were between 30-60 years of age, and 40.4% were female. We found that a total of 8.5% of the total respondents had been infected with the SARS-CoV-2 virus. Our survey revealed that out of 623 volunteers, 317 reported various side-effects after taking the vaccine, which is about 50.88% of the total participants. The majority of participants (37.07%, 231/623) reported swelling and pain at the injection site and fever (25.84%, 162/623); these were some of the common localized and generalized symptoms after the COVID-19 vaccine administration. CONCLUSION: The side-effects reported after receiving the Oxford-AstraZeneca vaccine (Covishield) are similar to those reported in clinical trials, demonstrating that the vaccines have a safe therapeutic window. Moreover, further research is needed to determine the efficacy of existing vaccines in preventing SARS-CoV-2 infections or after-infection hospitalization.

Diagnostic challenges and Gene-Xpert utility in detecting Mycobacterium tuberculosis among suspected cases of Pulmonary tuberculosis.

The incidence of pulmonary tuberculosis (PTB) can be reduced by preventing transmission with rapid and precise case detection and early treatment. The Gene-Xpert MTB/RIF assay is a useful tool for detecting Mycobacterium tuberculosis (MTB) with rifampicin resistance within approximately two hours by using a nucleic acid amplification technique. This study was designed to reduce the underdiagnosis of smear-negative pulmonary TB and to assess the clinical and radiological characteristics of PTB patients. This cross-sectional study included 235 participants who went to the Luyang primary health care clinic from September 2016 to June 2017. The demographic data were analyzed to investigate the association of patient gender, age group, and ethnicity by chi-square test. To assess the efficacy of the diagnostic test, the sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy were calculated. The area under the curve for sputum for both AFB and gene-Xpert was analyzed to compare their accuracy in diagnosing TB. In this study, TB was more common in males than in females. The majority (50.71%) of the cases belonged to the 25-44-year-old age group and the Bajau ethnicity (57.74%). Out of 50 pulmonary TB cases (smear-positive with AFB staining), 49 samples were positive according to the Gene-Xpert MTB/RIF assay and was confirmed by MTB culture. However, out of 185 smear-negative presumptive cases, 21 cases were positive by Gene-Xpert MTB/RIF assay in that a sample showed drug resistance, and these results were confirmed by MTB culture, showing resistance to isoniazid. In comparison to sputum for AFB, Gene-Xpert showed more sensitivity and specificity with almost complete accuracy. The additional 21 PTB cases detection from the presumptive cases by GeneXpert had significant impact compared to initial observation by the routine tests which overcame the diagnostic challenges and ambiguities.

Comprehensive care of a patient with obesity of BMI >70 kg/m2.

A 31-year-old woman, with a body mass index of 70.31 kg/m2, presented with progressive worsening of dyspnoea for 3 days. She had multiple comorbidities, including obesity hypoventilation syndrome. The patient developed type II respiratory failure with respiratory acidosis along with multiorgan failure. She was intubated and put on a mechanical ventilator and treated with intravenous diuretics, subcutaneous low-molecular-weight heparin and other supportive measures. Later, she was on noninvasive, continuous positive airway pressure ventilation overnight. She was prescribed a very-low-calorie diet along with physiotherapy and exercise. The patient underwent bariatric surgery 2 months after resolution of acute illness. Ten months after surgery, her body weight reduced from 180 kg to 121 kg, and her general condition improved. Successful management before and after surgical intervention depends on multidisciplinary teamwork, which includes the dietician, physiotherapist, endocrinologist, pulmonologist, nursing care and other supportive care.

Analysis of aceclofenac and bovine serum albumin interaction using fluorescence quenching method for predictive, preventive, and personalized medicine.

BACKGROUND: The study of the interaction of a drug with plasma protein is very important because drug-protein binding plays an important role in determination of pharmacological and toxicological properties of drugs. Our study was designed to investigate the interaction between aceclofenac and bovine serum albumin (BSA) using fluorescence spectroscopy at different temperatures (298 and 308 K). METHODS: Fluorescence spectroscopy was used to carry out the study. Fluorescence quenching constant was determined from Stern-Volmer equation. Van't Hoff equation was used to determine the thermodynamic parameters such as free energy (ΔG), enthalpy (ΔH), and entropy (ΔS). RESULTS: The experimental data showed that the quenching of BSA by aceclofenac was due to a formation of a BSA-aceclofenac complex with probable involvement of both tryptophan and tyrosine residues of BSA. Dynamic quenching was shown for BSA by aceclofenac at the experimental conditions. The values of thermodynamic parameters indicated that the hydrophobic forces played major roles for BSA-aceclofenac complexation. The binding number (n) was found to be ≈1 indicating that 1 mol of BSA bound with 1 mol of aceclofenac. The binding affinity of aceclofenac to BSA was calculated at different temperatures. It was shown that the binding constant decreased with increasing temperatures indicating that stability of the BSA-aceclofenac complex decreased with increasing temperatures. CONCLUSIONS: The interaction of aceclofenac with BSA was successfully explored using a fluorescence spectroscopic technique.

Real-time quantitative PCR assay on bacterial DNA: In a model soil system and environmental samples.

Real-time quantitative PCR (RTQ-PCR) was used to quantify the bacterial target DNA extracted by three commonly used DNA extraction protocols (bead mill homogenization, grinding in presence of liquid nitrogen and hot detergent SDS based enzymatic lysis). For the purpose of our study, pure culture of Bacillus cereus (model organism), sterilized soil seeded with a known amount of B. cereus (model soil system) and samples from woodland and grassland (environmental samples) were chosen to extract DNA by three different protocols. The extracted DNA was then quantified by RTQ-PCR using 16S rDNA specific universal bacterial primers. The standard curve used for the quantification by RTQ-PCR was linear and revealed a strong linear relationship (r(2)=0.9968) with a higher amplification efficiency, e5=1.02. High resolution gel electrophoresis was also carried out to observe the effect of these extraction methods on diversity analysis. For the model soil system, the liquid nitrogen method showed the highest target DNA copy number (1.3 x 10(9) copies/microl). However, for both the environmental samples, the bead beating method was found to be suitable on the basis of the high target DNA copy numbers (5.38 x 10(9) and 4.01 x 10(8) copies/ml for woodland and grassland respectively), high yield (6.4 microg/g and 1.76 microg/g of soil for woodland and grassland respectively) and different band patterns on high resolution gel electrophoresis suggesting an overall high extraction efficiency. This difference in the extraction efficiency between the model soil system and environmental samples may be attributed to different affinity of seeded and native DNA to soil particles.

Interference of contaminating DNA in the quantification of a toluene-induced tod gene in Pseudomonas putida.

To determine the extent of interference of co-extracted DNA contamination in the quantification of the tod gene transcript, two different concentrations of RNA (high, 500 ng/microl; low, 250 ng/microl) from a toluene-induced culture of Pseudomonas putida were treated with different amounts of DNase (2, 4, 6 and 8 U) and incubated for 30 and 60 min. The highly sensitive and reproducible TaqMan system was used to quantify the transcript of the tod gene, the tod gene in contaminating DNA and the 16S rRNA gene in DNase-treated RNA samples. For the high RNA concentration, the shorter incubation time (30 min) lowered the level of contaminating DNA as evidenced by the presence of 2.5 x 10(6) copies of the tod gene before treatment to 1.4 x 10(5) copies/microl (8 U), whereas, irrespective of the DNase units used, the longer incubation time (60 min) considerably lowered the level of DNA contamination (2.5 x 10(6) to 6.5 x 10(2) copies of the tod gene/microl). However, for the low RNA concentration, DNase treatment was found to be equally effective in lowering the level of contaminating DNA (10(6) to 10(2) copies of the tod gene/mu), irrespective of the incubation time and the amount of DNase used. Although the results of gel electrophoresis of conventional PCR amplification of the low RNA concentration revealed the absence of the target gene in contaminating DNA, the results of the TaqMan PCR indicated that a very low amount of contaminating DNA (less than 10(3) copies of the tod gene/mul) was still present in RNA samples, even after the DNase treatment. The number of copies of the tod gene transcript in RNA samples did not show any marked variation because of the DNase treatment. However, the proportion of contaminating DNA in RNA samples considerably decreased due to the treatment (0.01 to 0.000001). Furthermore, these results suggested that the extent of the removal of contaminating DNA from RNA samples depends on the concentration of RNA, the amount of DNase used and the incubation time. It is also suggested that the copies of the catabolic genes in contaminating DNA have to be quantified along with the target genes in RNA samples to have a more accurate quantification of the target genes for better understanding of their roles in many microbial processes.

Quantitative measurement of fungal DNA extracted by three different methods using real-time polymerase chain reaction.

Quantification of fungal populations in the environment is important for gaining a better understanding of various microbial processes. Recently, the development of real-time quantitative PCR (RTQ-PCR) has eliminated the variability associated with conventional quantitative PCR, thereby allowing the routine and reliable quantification of PCR products. Thus, in this present study, RTQ-PCR was used to quantify the fungal target DNA extracted by three commonly used DNA extraction protocols (bead mill homogenization, grinding in the presence of liquid nitrogen, and hot detergent SDS based enzymatic lysis combined with bead beating) to determine the suitability of the quantification of target DNA. For the purpose of this study, pure culture of Aspergillus fumigatus (model organism), sterilized soil seeded with a known amount ofA. fumigatus (model soil system), and woodland and grassland soil samples (environmental samples) were chosen to extract DNA by the above three different protocols. The extracted DNA was then quantified by spectroscopy and a RTQ-PCR system. 18S rDNA specific universal fungal primers were used to quantify the target part and then amplification products were verified by agarose gel electrophoresis. Standard curves used for the quantification by RTQ-PCR revealed strong linear relationships (R2=0.9994 for the primer pair NS1 and NS2 and 0.9938 for the primer pair nu-SSU-0817 and nu-SSU-1196) with a higher amplification efficiency, e=0.983 for the primer pair NS1 and NS2 and 0.956 for the primer pair nu-SSU-0817 and nu-SSU-1196. Although for pure culture the hot detergent SDS based enzymatic lysis combined with bead beating method showed the highest target DNA copy number (1.5 x 10(9) copies/microl), for the model soil system and both environmental samples the bead beating method was found to be suitable on the basis of the high target DNA copy numbers (6.16 x 10(8) and 2.7 x 10(8) copies/microl for woodland and grassland, respectively), high DNA yield (6.4 microg/g and 1.8 microg/g of soil for woodland and grassland, respectively), and high recovery on the basis of the target DNA copy number (39.2%), suggesting an overall high extraction efficiency.