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Fulltext Bioactivity-Guided Fractionation and Identification of Antidiabetic Compound of Syzygium polyanthum (Wight.)'s Leaf Extract in Streptozotocin-Induced Diabetic Rat Model

Widyawati T, Yusoff NA, Bello I, Asmawi MZ, Ahmad M

Molecules, 2022 Oct 12;27(20).
PMID: 36296407 DOI: 10.3390/molecules27206814

(1) Background: An earlier study on the hypoglycemic activity of S. polyanthum (Wight.) leaf methanol extract identified squalene as the major chemical compound. The present study was conducted to assess the hypoglycemic effect of fractions and subfractions of the methanol extract of S. polyanthum compared to the squalene using a bioassay-guided in vivo study. (2) Methods: The methanol extract was fractionated using the liquid−liquid fractionation method. Streptozotocin-induced type 1 diabetic rat was used to study the hypoglycemic effect. (3) Results: The findings showed that chloroform fraction significantly (p < 0.05) lowered blood glucose levels of diabetic rats as compared to the control. Further fractionation of chloroform fraction yielded subfraction-1 and -2, whereby subfraction-1 exhibited a higher blood-glucose-lowering effect. The lipid profile test showed that the total cholesterol level of subfraction-1 and squalene-treated groups decreased significantly (p < 0.05). An immunohistochemistry study revealed that none of the treatments regenerated pancreatic β-cells. Gas chromatography−mass spectrophotometer analysis identified the presence of squalene in the active methanol extract, chloroform fraction, and subfraction-1. In silico analysis revealed a higher affinity of squalene against protein receptors that control lipid metabolism than metformin. (4) Conclusions: Data obtained from the present work suggested the crude methanol extract exerted the highest hypoglycemic effect compared to fraction, subfraction, and squalene, confirming synergistic effect may be responsible for the hypoglycemic activity of S. polyanthum.

Matched MeSH terms: Chloroform
Fulltext Assessment of RNA extraction protocols from cladocerans

Razak MR, Aris AZ, Md Yusoff F, Yusof ZNB, Kim SD, Kim KW

PLoS One, 2022;17(4):e0264989.
PMID: 35472091 DOI: 10.1371/journal.pone.0264989

The usage of cladocerans as non-model organisms in ecotoxicological and risk assessment studies has intensified in recent years due to their ecological importance in aquatic ecosystems. The molecular assessment such as gene expression analysis has been introduced in ecotoxicological and risk assessment to link the expression of specific genes to a biological process in the cladocerans. The validity and accuracy of gene expression analysis depends on the quantity, quality and integrity of extracted ribonucleic acid (RNA) of the sample. However, the standard methods of RNA extraction from the cladocerans are still lacking. This study evaluates the extraction of RNA from tropical freshwater cladocerans Moina micrura using two methods: the phenol-chloroform extraction method (QIAzol) and a column-based kit (Qiagen Micro Kit). Glycogen was introduced in both approaches to enhance the recovery of extracted RNA and the extracted RNA was characterised using spectrophotometric analysis (NanoDrop), capillary electrophoresis (Bioanalyzer). Then, the extracted RNA was analysed with reverse transcription polymerase chain reaction (RT-PCR) to validate the RNA extraction method towards downstream gene expression analysis. The results indicate that the column-based kit is most suitable for the extraction of RNA from M. micrura, with the quantity (RNA concentration = 26.90 ± 6.89 ng/μl), quality (A260:230 = 1.95 ± 0.15, A280:230 = 1.85 ± 0.09) and integrity (RNA integrity number, RIN = 7.20 ± 0.16). The RT-PCR analysis shows that the method successfully amplified both alpha tubulin and actin gene at 33-35 cycles (i.e. Ct = 32.64 to 33.48). The results demonstrate that the addition of glycogen is only suitable for the phenol-chloroform extraction method. RNA extraction with high and comprehensive quality control assessment will increase the accuracy and reliability of downstream gene expression, thus providing more ecotoxicological data at the molecular biological level on other freshwater zooplankton species.

Matched MeSH terms: Chloroform*
Antibacterial and antibiotic potentiating capabilities of extracts isolated from Burkillanthus malaccensis, Diospyros hasseltii and Cleisthanthus bracteosus against human pathogenic bacteria

Kathirvalu G, Chandramathi S, S A A, Atiya N, Begum S, Christophe W, et al.

Trop Biomed, 2023 Jun 01;40(2):152-159.
PMID: 37650400 DOI: 10.47665/tb.40.2.004

Antibiotics which once a boon in medicine and saved millions of lives are now facing an ever-growing menace of antibacterial resistance, which desperately needs new antibacterial drugs which are innovative in chemistry and mode of action. For many years, the world has turned to natural plants with antibacterial properties to combat antibiotic resistance. On that basis, we aimed to identify plants with antibacterial and antibiotic potentiating properties. Seventeen different extracts of 3 plants namely Burkillanthus malaccensis, Diospyros hasseltii and Cleisthanthus bracteosus were tested against multi-drug resistant Acinetobacter baumannii, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Methicillinresistant Staphylococcus aureus (MRSA) and methicillin-susceptible Staphylococcus aureus (MSSA). Antibacterial activity of hexane, methanol and chloroform extracts of bark, seed, fruit, flesh and leaves from these plants were tested using, disk diffusion assay, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) assays. Antibiotic potentiating capabilities were tested using time-kill assay. B. malaccensis fruit chloroform extract showed the biggest zone of inhibition against MRSA (13.00±0.0 mm) but C. bracteosus bark methanol extract showed the biggest inhibition zone against MSSA (15.33±0.6 mm). Interestingly, bark methanol extract of C. bracteosus was active against MRSA (8.7±0.6 mm), MSSA (7.7±0.6 mm) (Gram-positive) and A. baumannii (7.7±0.6 mm) (Gram-negative). Overall, the leaf methanol and bark methanol extract of C. bracteosus warrants further investigation such as compound isolation and mechanism of action for validating its therapeutic use as antibiotic potentiator importantly against MRSA and A. baumannii.

Matched MeSH terms: Chloroform/pharmacology
Fulltext Liver protective effect of chloroform extract of Bauhinia purpurea leaves is attributed partly to its antioxidant action and the presence of flavonoids

Zakaria ZA, Sahmat A, Hizami Azmi A, Zainol ASN, Omar MH, Balan T, et al.

Pharm Biol, 2023 Dec;61(1):1152-1161.
PMID: 37559390 DOI: 10.1080/13880209.2023.2241510

CONTEXT: Bauhinia purpurea L. (Fabaceae) is used in the Ayurvedic system to treat various oxidative-related ailments (e.g., wounds, ulcers etc.). Therefore, it is believed that the plant also has the potential to alleviate oxidative-related liver damage.
OBJECTIVE: This study elucidates the hepatoprotective activity of chloroform extract of B. purpurea leaves (CEBP) in paracetamol (PCM)-induced liver injury (PILI) rats.
MATERIALS AND METHODS: Male Sprague-Dawley rats (n = 6) were pre-treated once daily (p.o.) with CEBP (50-500 mg/kg) for seven consecutive days before being administered (p.o.) a hepatotoxic agent, 3 g/kg PCM. Liver enzyme levels were determined from the collected blood, while the collected liver was used to determine the activity of endogenous antioxidant enzymes and for histopathological examination. CEBP was also subjected to radical scavenging assays and phytochemical analysis.
RESULTS: CEBP significantly (p

Matched MeSH terms: Chloroform/pharmacology
Encapsulation of Dicranopteris linearis extract using cellulose microparticles for antiulcer medication

Ahmad AA, Kasim KF, Gopinath SCB, Anbu P, Sofian-Seng NS

Int J Biol Macromol, 2023 Dec 31;253(Pt 2):126795.
PMID: 37689304 DOI: 10.1016/j.ijbiomac.2023.126795

Dicranopteris linearis (DL) is a fern in the Gleicheniaceae family, locally known as resam by the Malay community. It has numerous pharmacological benefits, with antiulcer and gastroprotective properties. Peptic ulcer is a chronic and recurring disease that significantly impacts morbidity and mortality, affecting nearly 20 % of the world's population. Despite the effectiveness of peptic ulcer drugs, there is no perfect treatment for the ailment. Encapsulation is an advanced technique that can treat peptic ulcers by incorporating natural sources. This work aims to encapsulate DL extract using different types of cellulose particles by the solvent displacement technique for peptic ulcer medication. The extract was encapsulated using methyl cellulose (MC), ethyl cellulose (EC), and a blend of ethyl methyl cellulose through a dialysis cellulose membrane tube and freeze-dried to yield a suspension of the encapsulated DL extracts. The microencapsulated methyl cellulose chloroform extract (MCCH) has a considerably greater level of total phenolic (84.53 ± 6.44 mg GAE/g), total flavonoid (84.53 ± 0.54 mg GAE/g), and antioxidant activity (86.40 ± 0.63 %). MCCH has the highest percentage of antimicrobial activity against Escherichia coli (2.42 ± 107 × 0.70 CFU/mL), Bacillus subtilis (5.21 ± 107 × 0.90 CFU/mL), and Shigella flexneri (1.25 ± 107 × 0.66 CFU/mL), as well as the highest urease inhibitory activity (50.0 ± 0.21 %). The MCCH particle size was estimated to be 3.347 ± 0.078 μm in diameter. It has been proven that DL elements were successfully encapsulated in the methyl cellulose polymer in the presence of calcium (Ca). Fourier transform infrared (FTIR) analysis indicated significant results, where the peak belonging to the CO stretch of the carbonyl groups of methyl cellulose (MC) shifted from 1638.46 cm-1 in the spectrum of pure MC to 1639.10 cm-1 in the spectrum of the MCCH extract. The shift in the wavenumbers was due to the interactions between the phytochemicals in the chloroform extract and the MC matrix in the microcapsules. Dissolution studies in simulated gastric fluid (SGF) and model fitting of encapsulated chloroform extracts showed that MCCH has the highest EC50 of 6.73 ± 0.27 mg/mL with R2 = 0.971 fitted by the Korsmeyer-Peppas model, indicating diffusion as the mechanism of release.

Matched MeSH terms: Chloroform