Optimization of nutrient parameters for lovastatin production by <Emphasis Type="Italic">Monascus purpureus</Emphasis> MTCC 369 under submerged fermentation using response surface methodology

Lovastatin, an inhibitor of HMG-CoA reductase, was produced by submerged fermentation using Monascus purpureus MTCC 369. Five nutritional parameters screened using Plackett–Burman experimental design were optimized by Box–Behnken factorial design of response surface methodology for lovastatin production in shake flask cultures. Maximum lovastatin production of 351 mg/l were predicted in medium containing 29.59 g/l dextrose, 3.86 g/l NH₄Cl, 1.73 g/l KH₂PO₄, 0.86 g/l MgSO₄·7H₂O, and 0.19 g/l MnSO₄·H₂O using response surface plots and point prediction tool of DESIGN EXPERT 7.0 (Statease, USA) software.     

Large Scale Spatial Variability of Accumulated Cadmium in the Wheat Farm Grains

Cadmium (Cd) accumulation in edible crops is undesirable due to its hazardous influences on human health. The objectives of this study were: i) to evaluate the spatial variability of grain Cd and its relationships with soil properties in 4000 km² wheat farms; ii) to evaluate the effect of wheat cultivar on the soil properties vs. grain Cd relationships. A number of 255 soil (0–20 cm) and grain samples were taken and Cd concentrations in grain samples and some soil properties were measured. Grain Cd concentrations in 95 percent of the samples exceeded the threshold of 0.2 mg kg^?1. Durum wheat had more potential to accumulate Cd in grain (0.76 mg kg^?1) than bread (0.69 mg kg^?1). There was significant (p < 0.01) correlation between grain Cd and organic carbon (r = 0.66), CEC (r = 0.77) and DTPA-extractable Cd (p < 0.05) (r = 0.57) of the soils. Greater Pearson coefficient values for durum wheat showed that, in the studied calcareous soils, organic carbon, CEC, Cd-DTPA had more effects on durum wheat than bread wheat cultivar. The obtained Kriging map of grain Cd identified three hotspots at the east (durum wheat cultivation), the west (intensive irrigated wheat farms), and south (wheat farms around petrochemical industries) of the region. Agricultural mismanagement due to overusing P-fertilizers increased Cd concentration in the topsoils and grains of wheat farms in the study area.     

Effects of Cadmium,Copper, Lead,and Zinc Contamination on Metal Accumulation by Safflower and Wheat

Accumulation of heavy metals (HMs) in cultivated soils is a continuing environmental problem in many parts of the world. An increase in HM concentration can enhance uptake of toxic metals by crops and enter the human food chain. In this study, the uptake behavior of wheat and safflower was evaluated in a calcareous soil by using 12 undisturbed columns in which half were artificially contaminated. Heavy metals in the form of CdCl₂ (15 mg Cd kg^{? 1}), CuSO₄ (585 mg Cu kg^{? 1}), Pb(NO₃)₂ (117 mg Pb kg^{? 1}), and ZnCl₂ (1094 mg Zn kg^{? 1}) were sprayed on the soil surface and completely mixed in the top 10 cm. The background total concentrations of Cd, Cu, Pb and Zn were 1.6, 29.5, 17.5 and 61.2 mg kg^{? 1}, respectively. After metal application, half of the columns (3 contaminated and 3 uncontaminated) were sown with wheat (Triticum aestivum) and the other half with safflower (Carthamus tinctorious) and grown for 74 days until maturity. After harvesting, soil columns were cut into 10-cm sections and analyzed for HNO₃- and DTPA-extractable metal concentrations. Metal concentrations were also measured in different plant tissues. The results showed that artificial contamination of topsoil decreased the transpiration rate of wheat by 12% and that of safflower by 6%. In contaminated columns, Cd, Cu, Pb, and Zn accumulation in wheat shoot was greater by 8.0-, 1.9-, 3.0-, and 2.1-fold than the control, respectively. Accordingly, these numbers were 46.0-, 1.3-, 1.7-, and 1.6-fold in safflower shoot. Soil contamination with HMs resulted in a 55% decrease in shoot dry matter yield of wheat while it had no significant effect on shoot dry matter of safflower. The normalized water consumption for safflower was therefore not affected by metal contamination (≈ 13 mm H₂O g^{? 1} of dry weight for all safflower and uncontaminated wheat treatments), while contaminated wheat was much less water efficient at about 27 mm H₂O g^{? 1} dry weight. It was concluded that although artificial contamination had a negative effect on wheat growth, it did not affect safflower's normal growth and water efficiency.     

The Role of Rac1 in the Growth Cone Dynamics and Force Generation of DRG Neurons

We used optical tweezers, video imaging, immunocytochemistry and a variety of inhibitors to analyze the role of Rac1 in the motility and force generation of lamellipodia and filopodia from developing growth cones of isolated Dorsal Root Ganglia neurons. When the activity of Rac1 was inhibited by the drug EHop-016, the period of lamellipodia protrusion/retraction cycles increased and the lamellipodia retrograde flow rate decreased; moreover, the axial force exerted by lamellipodia was reduced dramatically. Inhibition of Arp2/3 by a moderate amount of the drug CK-548 caused a transient retraction of lamellipodia followed by a complete recovery of their usual motility. This recovery was abolished by the concomitant inhibition of Rac1. The filopodia length increased upon inhibition of both Rac1 and Arp2/3, but the speed of filopodia protrusion increased when Rac1 was inhibited and decreased instead when Arp2/3 was inhibited. These results suggest that Rac1 acts as a switch that activates upon inhibition of Arp2/3. Rac1 also controls the filopodia dynamics necessary to explore the environment.     

pH-Dependent Solubility and Dissolution Behavior of Carvedilol—Case Example of a Weakly Basic BCS Class II Drug

The objective of this study was to investigate the pH-dependent solubility and dissolution of weakly basic Biopharmaceutical Classification Systems (BCS) class II drugs, characterized by low solubility and high permeability, using carvedilol, a weak base with a pK _a value of 7.8, as a model drug. A series of solubility and in vitro dissolution studies was carried out using media that simulate the gastric and intestinal fluids and cover the physiological pH range of the GI from 1.2 to 7.8. The effect of ionic strength, buffer capacity, and buffer species of the dissolution media on the solubility and dissolution behavior of carvedilol was also investigated. The study revealed that carvedilol exhibited a typical weak base pH-dependent solubility profile with a high solubility at low pH (545.1–2591.4 μg/mL within the pH range 1.2–5.0) and low solubility at high pH (5.8–51.9 μg/mL within the pH range 6.5–7.8). The dissolution behavior of carvedilol was consistent with the solubility results, where carvedilol release was complete (95.8–98.2% released within 60 min) in media simulating the gastric fluid (pH 1.2–5.0) and relatively low (15.9–86.2% released within 240 min) in media simulating the intestinal fluid (pH 6.5–7.8). It was found that the buffer species of the dissolution media may influence the solubility and consequently the percentage of carvedilol released by forming carvedilol salts of varying solubilities. Carvedilol solubility and dissolution decreased with increasing ionic strength, while lowering the buffer capacity resulted in a decrease in carvedilol solubility and dissolution rate.     

Plant-derived VLP: a worthy platform to produce vaccine against SARS-CoV-2

Biotechnology Letters - After its emergence in late 2019 SARS-CoV-2 was declared a pandemic by the World Health Organization on 11 March 2020 and has claimed more than 2.8 million lives. There has...     

Side reactions in the SPPS of Cys-containing peptides

Alkylation of sensitive amino acids during synthesis of biologically important peptides is a common and well-documented problem in Fmoc-based strategy. Herein, we probed for the first time an unexpected S-alkylation of Cys-containing peptides that occur during the final TFA cleavage of peptides from the Wang solid support. Through a battery of approaches (NMR, UV and LC–MS) the formed by-product was assigned as the alkylation of the cysteine sulfydryl group by the p-hydroxyl benzyl group derived from the acidic Wang linker decomposition. Factors affecting this side reaction were monitored and a protocol that minimizes the presence of the by-product is reported.     

An appraisal on synthetic and pharmaceutical perspectives of pyrazolo[4,3-d]pyrimidine scaffold

Pyrazolo[4,3-d]pyrimidine, a fused heterocycle bearing pyrazole and pyrimidine portions has gained a significant attention in the field of bioorganic and medicinal chemistry. Pyrazolo[4,3-d]pyrimidine derivatives have demonstrated numerous pharmacological activities particularly, anti-cancer, anti-infectious, phosphodiesterase inhibitors, adenosine antagonists and cytokinin antagonists etc. This review extensively unveils the synthetic and pharmacological diversity with special emphasis on structural variations around pyrazolo[4,3-d]pyrimidine scaffold. This endeavour has thus uncovered the medicinal worthiness of pyrazolo[4,3-d]pyrimidine framework. To the best of our knowledge this review is the first compilation on synthetic, medicinal and structure activity relationship (SAR) aspects of pyrazolo[4,3-d]pyrimidines since 1956.     

Design and synthesis of novel thiadiazole-thiazolone hybrids as potential inhibitors of the human mitotic kinesin Eg5

A novel series of 1,3,4-thiadiazole-thiazolone hybrids 5a–v were designed, synthesized, characterized, and evaluated against the basal and the microtubule (MT)-stimulated ATPase activity of Eg5. From the evaluated derivatives, 5h displayed the highest inhibition with an IC₅₀ value of 13.2?µM against the MT-stimulated Eg5 ATPase activity. Similarly, compounds 5f and 5i also presented encouraging inhibition with IC₅₀ of 17.2?µM and 20.2?µM, respectively. A brief structure–activity relationship (SAR) analysis indicated that 2-chloro and 4-nitro substituents on the phenyl ring of the thiazolone motif contributed significantly to enzyme inhibition. An in silico molecular docking study using the crystal structure of Eg5 further supported the SAR and reasoned the importance of crucial molecular protein–ligand interactions in influencing the inhibition of the ATPase activity of Eg5. The magnitude of the electron-withdrawing functionalities over the hybrids and the critical molecular interactions contributed towards higher in vitro potency of the compounds. The drug-like properties of the synthesized compounds 5a–v were also calculated based on the Lipinski’s rule of five and in silico computation of key pharmacokinetic parameters (ADME). Thus, the present work unveils these hybrid molecules as novel Eg5 inhibitors with promising drug-like properties for future development.