GC-MS ANALYSIS PROFILE AND BIOACTIVE COMPONENTS OF FLOWERS OF Bergenia ciliata, A WEED OF ROCK CREVICES

Bergenia ciliata is a weed of hilly areas of Pakistan that generally grows in rock crevices. In the present study, flowers of this weeds were collected from Murree. The dried flowers were extracted in methanol and subjected to GC-MS analysis that showed 7 compounds in it. The predominant compound was hexanedioic acid, bis(2-ethylhexyl) ester (48.88%) followed by γ-sitosterol (22.56%). Moderately occurring compound was cyclohexane, 1,3,5-triphenyl- (12.87%). The remaining compounds namely n-hexadecanoic acid (4.97%), pentadecanoic acid, 14-methyl-, methyl ester (3.77%), 9,12-octadecadienoic acid, methyl ester, (Z,Z)- (2.94%) and 9,12,15-octadecatrienoic acid, methyl ester, (Z,Z,Z)- (2.92%) were categorized as less abundant compounds. Literature survey indicated that these compounds possess antimicrobial, larvicidal, anti-inflammatory, anticancer and/or antidiabetic properties.

Tunnel engineering for modulating the substrate preference in cytochrome P450BsβHI

An active site is normally located inside enzymes, hence substrates should go through a tunnel to access the active site. Tunnel engineering is a powerful strategy for refining the catalytic properties of enzymes. Here, P450BsβHI (Q85H/V170I) derived from hydroxylase P450Bsβ from Bacillus subtilis was chosen as the study model, which is reported as a potential decarboxylase. However, this enzyme showed low decarboxylase activity towards long-chain fatty acids. Here, a tunnel engineering campaign was performed for modulating the substrate preference and improving the decarboxylation activity of P450BsβHI. The finally obtained BsβHI-F79A variant had a 15.2-fold improved conversion for palmitic acid; BsβHI-F173V variant had a 3.9-fold improved conversion for pentadecanoic acid. The study demonstrates how the substrate preference can be modulated by tunnel engineering strategy.

Plasma Free Fatty Acids and Metabolic Effect in Type 2 Diabetes, an Ancillary Study from a Randomized Clinical Trial

Most nutrition studies looking at the association of food with cardiometabolic markers rely on food frequency questionnaires, which are prone to recall bias. Pentadecanoic acid, heptadecanoic acid and trans-palmitoleic acid are fatty acids that are not synthesized endogenously but are obtained from the diet, particularly dairy, making them reasonable biomarkers of dairy consumption. We investigated the association of dairy fatty acid biomarkers with glycated hemoglobin (HbA1c) and cardiovascular risk factors in type 2 diabetes (T2D). In a clinical trial, 111 participants with T2D (age 58.5 ± 8.9 years, HbA1c 8.09 ± 0.96%) were randomized into three groups: a control group that maintained baseline dairy intake, a low-fat (LF) group that incorporated ≥3 servings/day of LF dairy and a high-fat (HF) group that incorporated ≥3 servings/day of HF dairy. We compared the fatty acids (FA) composition between the three groups at 24 weeks. Pentadecanoic acid and trans-palmitoleic acid increased in the HF group by 14.1% ± 5.4% and 17.5% ± 5.1%, respectively, but not in the control and LF groups (p = 0.0474 and p= 0.0025 for group-by-time interaction, respectively). Those increases were positively associated with changes in total cholesterol, very-low-density lipoprotein cholesterol VLDL-C and triglycerides but were not associated with changes in HbA1c from baseline to 24 weeks. These results suggest that the intervention was successful and that participants were likely compliant, which supports the validity of the main trial.

Behavioural and antennal responses of Aedes aegypti (l.) (Diptera: Culicidae) gravid females to chemical cues from conspecific larvae

Mass trapping of gravid females represents one promising strategy for the development of sustainable tools against Aedes aegypti. However, this technique requires the development of effective odorant lures that can compete with natural breeding sites. The presence of conspecific larvae has been shown to stimulate oviposition. Hence, we evaluated the role of four major molecules previously identified from Ae. aegypti larvae (isovaleric, myristoleic, myristic [i.e. tetradecanoic], and pentadecanoic acids) on the oviposition of conspecific females, as well as their olfactory perception to evaluate their range of detection. Using flight cage assays, the preference of gravid females to oviposit in water that previously contained larvae (LHW) or containing the four larval compounds was evaluated. Then, compounds and doses inducing the highest stimulation were challenged for their efficacy against LHW. Only isovaleric acid elicited antennal response, suggesting that the other compounds may act as taste cues. Pentadecanoic acid induced significant oviposition stimulation, especially when dosed at 10 ppm. Myristoleic acid and isovaleric acid deterred oviposition at 10 and 100 ppm, while no effect on oviposition was observed with myristic acid irrespectively of the dose tested. When the four compounds were pooled to mimic larvae’s chemical signature, they favored oviposition at 1 ppm but negatively affected egg-laying at higher concentrations. When properly dosed, pentadecanoic acid and the blend of compounds may be promising lures for ovitraps as they could compete with LHW. Due to their low volatility, their effect should be further evaluated under field conditions, in addition with long-range attractants for developing effective tools against gravid females.

Tunnel Engineering for Modulating the Substrate Preference in Decarbonylase P450BsβHI

An active site normally locates inside of enzymes, substrates should go through the tunnel to access the active site. Tunnel engineering is a powerful strategy for refining the catalytic properties of enzymes. Here, P450 Bsβ HI (Q85H/ V170I) derived from hydroxylase P450 Bsβ from Bacillus subtilis was chosen as study model, which is reported as a potential decarbonylase. However, this enzyme showed low decarboxylase activity towards long-chain fatty acids. Here, a tunnel engineering campaign was performed for modulating the substrate preference and improving the decarbonylase activity of P450 Bsβ HI. The finally obtained BsβHI-F79A variant had a 15.2-fold improved conversion for palmitic acid; BsβHI-F173V variant had a 3.9-fold improved conversion for pentadecanoic acid. The study demonstrates how the substrate preference can be modulated by tunnel engineering strategy.

Tunnel Engineering for Modulating The Substrate Preference in Decarbonylase P450BsβHI

An active site normally locates inside of enzymes, substrates should go through the tunnel to access the active site. Tunnel engineering is a powerful strategy for refining the catalytic properties of enzymes. Here, P450BsβHI (Q85H/ V170I) derived from hydroxylase P450Bsβ from Bacillus subtilis was chosen as study model, which is reported as a potential decarbonylase. However, this enzyme showed low decarboxylase activity towards long-chain fatty acids. Here, a tunnel engineering campaign was performed for modulating the substrate preference and improving the decarbonylase activity of P450BsβHI. The finally obtained BsβHI-F79A variant had a 15.2-fold improved conversion for palmitic acid; BsβHI-F173V variant had a 3.9-fold improved conversion for pentadecanoic acid. The study demonstrates how the substrate preference can be modulated by tunnel engineering strategy.

Proximate composition and GC-MS analysis of ethanol extract of Solanum spirale Roxb.

Solanum spirale Roxb. is abundantly grown in eastern Himalayan regions. The shoot is used as a vegetable as well as medicine to control high blood pressure. The present study was carried out to profile the phytochemical compositions, mineral contents and proximate composition of the Solanum spirale shoot. GC-MS was used for the identification of phytochemicals and methods described in standard protocols were used to study minerals and proximate composition. A total of 40 phytochemicals were recorded from the ethanol extract. Mineral contents of the sample 0.115 (Fe), 0.07 (Mn), 0.015 (Cu), 0.040 (Zn), 2.25 (Mg), 3.08 (Na) and 16.7 (K) mg/g while proximate composition were 76.25± 0.093 % (moisture)3.82±0.26 % (Carbohydrate), 12.54±0.08% (Total ash), 0.39±0.98% (Crude protein), 6.12±1.07 % (Crude fibre), 0.37±0.07 % (Crude fat). (Z,Z)-6,9-Cis-3,4-epoxy-nonadecadiene occupied highest area percentage in TIC peak report with 24.55%, followed by Pentadecanoic acid with 18.81 percent, with Pentadecane with 9.99%, Hexadecanoicacid, Ethyl ester with 8.64%, (2E)(7R,11R)-3,7,11,15-Tetramethylhexadec-2-en-1-ol with 7.75%. Useful phytocompounds related to health problems including anticancer, anti-inflammatory, antioxidant, antitumor, cardioprotective, hypocholesterolemic, increase zinc bioavailability, inhibit uric acid formation, antibacterial, anti-inflamatory, antioxidant etc. are major components and such findings advocate Solanum spirale as a nutraceutical herb.

Chemical profiling of two aromatic weeds, Cyathocline purpurea and Blumea lacera

In the present investigation chemical constituents of Cyathocline purpurea (Buch.-Ham. ex D.Don) Kuntze and Blumea lacera (Burm.f.) DC. (Family-Compositae) were studied by using gas chromatography coupled with mass spectrometry (GC/MS). These two weeds are small herbs and well known for its potent medicinal properties. Total 17 and 27 compounds were identified from C. purpurea and B. lacera respectively. The major constituents in both the extracts were pentadecanoic acid, 14-methyl-, methyl ester (30.56 %), cis-phytol (21.26 %), α- cadinol (7.87 %), γ-cadinene (7.13 %), neophytadiene (3.81 %) and α-cubebene (1.82 %). GC/MS analysis revealed the presence of various bioactive compounds such as fatty acids, sesquiterpenoids, phenols, etc. in the acetone extracts of both the plants. The identified compounds have various biological activities.