Genetic Profiling, Chemical Characterization and Biological Evaluation of Two Conyza Species Growing in Egypt.

The present report is a comparative investigation of two Conyza species growing wild in Egypt namely, Conyza dioscoridis (L.) Desf. and Conyza bonariensis (L.) Cronquist. It comprises a genetic and chemical characterization of the plants, as well as an evaluation of their biological activities. The DNA fingerprints of the two species were established based on a polymerase chain reaction (PCR) procedure using ten decamer primers. Further characterization of the plants was performed via determination of pharmacopoeial constants, phytochemical screening and estimation of phenolic content (total phenolics, tannins and flavonoids). The ethanol (70%) extracts of C. dioscoridis (EECD) and C. bonariensis (EECB) were subjected to acute toxicity study to determine their LD50; the anti-inflammatory, antimicrobial and cytotoxic activities were then evaluated. Screening for potential cytotoxicity was carried out both by Brine Shrimp Lethality Test and Sulphorodamine-B assay on three human cell lines viz., breast carcinoma (MCF7), colorectal carcinoma (HCT116) and cervical carcinoma (HELA) cell lines. The DNA profiling revealed a similarity index of 88.89% between the investigated species. The variability observed among the pharmacopoeial constants constitute a valuable differential criterion; the total ash, acid insoluble ash, water soluble ash and crude fiber values obtained for C. bonariensis exceeded (17, 5, 10 and 3.5%, respectively) those for C. dioscoridis; meanwhile, the moisture content was higher (10%) in the latter. The phytochemical screening of EECD and EECB revealed the presence of flavonoids, steroids, terpenoids and tannins in both species. Estimation of phenolic contents (total phenolics, tannins and flavonoids expressed as gallic acid, tannic acid and rutin equivalents, respectively) showed that EECD contains higher amounts of all these constituents when compared to EECB (1.17 vs. 0.96 mg/g, total phenolics; 2.43 vs. 1.83 mg/g, tannins and 0.62 vs. 0.29 mg/g, flavonoids). EECD and EECB were found to be safe (LD50 upto 0.5g/kg). Throughout evaluation of the antimicrobial activity against a set of microbial strains and potential cytotoxicity against MCF7, EECD appeared more efficient (MIC: 200-400 μg/ml and IC50: 2.97 μg/ml, respectively); meanwhile, the effect of EECB was more significant on HCT116 and HELA (IC50: 21 and 5.4 μg/ml, respectively). Results of in-vivo assessment of the anti-inflammatory activity showed that the inhibitory effect of EECD was more prominent than that of EECB (74.20% vs. 59.0%). However, the effect of the extracts was inversed in the Brine Shrimp Lethality test (30% vs. 40% lethality, respectively).

Estrogenic activity of Citrus medica L. leaves growing in Egypt.

The estrogenic activity of petroleum ether extract of Citrus medica L. leaves as well as the chemical constituents responsible for the biological activity was studied. The petroleum ether extract proved to retain high estrogenic activity in immature female rats. The extract was saponified and its components (saponifiable part 23% and the unsaponifiable matter 77%) were identified using GC/MS technique. The extract proved to be safe (LD50< 2g/kg.bw). Oral administration of petroleum ether extract of C. medica in ovariectomized immature female Wistar rats for 7 days in a dose of 400 mg/kg resulted in significant increase in the uterine weight (g) (1.7±0.11) when compared with ovariectomized control rats (1.3±0.07). GC/MS analysis of both saponifiable and unsaponifiable matters revealed the presence of thirty three components (28 hydrocarbons and 5 sterols) in the unsaponifiable fraction, the major hydrocarbon was n- Heneicosane (16.7%) while the major sterol was β-sitosterol (4.03%) and 15 components in the saponifiable matter it's major component was hexadecanoic acid (19.93%). As a conclusion petroleum ether extract of Citrus medica L. leaves possess a significant estrogenic activity.

Antiviral activity of freshwater algae.

Five freshwater algal species were isolated from Nile River and studied for their biological (cytotoxic and antiviral activity) in order to test their benefit in the Egyptian drinking water source. The algal species were isolated and identified as: Anabaena sphaerica, Chroococcus turgidus, Oscillatoria limnetica, and Spirulina platensis (blue – green algae, Cyanobacteria) and Cosmarium leave (green algae). They were cultivated using a Photobioreactor and purified using BG11 media. Twenty five grams of each of the five powdered algal species were extracted with MeOH till exhaustion to give five methanolic extracts for Anabaena sphaerica, Chroococcus turgidus, Oscillatoria limnetica, Spirulina platensis and Cosmarium leave respectively. The residues left were extracted with distilled H2O at 50oC to give five aqueous extracts respectively. The cytotoxicity of all the extracts was tested on Hep-2 cell line and their antiviral assays were tested on Adenovirus Type 40 as a preliminary testing. Nested PCR was carried out for confirmation of Adenovirus. Antialgal inhibitory effect on algal community was carried out. Results revealed that the non toxic concentrations for all the extracts were 2mg/ml and Spirulina platensis methanol and water extracts were active alga as antiviral (50% and 23.3% of reduction respectively).

Efficacy of Citrus reticulata and Mirazid in treatment of Schistosoma mansoni

This work has been carried out to investigate the effect of Schistosoma mansoni infection on mice livers after treatment with the ethanolic extract of Citrus reticulata root or the oleo-resin extract from Myrrh of Commiphora molmol tree (Mirazid), as a new antishistosomal drug. Marker enzymes for different cell organelles were measured; succinate dehydrogenase (SDH); lactate dehydrogenase (LDH) and its isoenzymes; glucose-6-phosphatase (G-6-Pase); acid phosphatase (AP) and 5'- nucleotidase. Liver function enzymes; aspartate aminotransferase (AST); alanine aminotransferase (ALT), and alkaline phosphatase (ALP) were also estimated. Parasitological studies through ova count and worm burden will also be taken into consideration. The results showed a marked reduction in SDH, LDH, AST, and ALT enzyme activities and a significant increase in G-6-Pase, AP, 5'- nucleotidase, and ALP after S. mansoni infection. A noticeable alteration in LDH subunits were also noticed. Treatment with C. reticulata or Mirazid improved all the previous enzyme activities with a noticeable reduction in ova count and worm burden.