Rol genes enhance content of artemisinin and other secondary metabolites in Shennong hybrid of Artemisia annua / 中草药·英文版

Objective: Artemisia annua is the chief source of artemisinin, a potent antimalarial agent, in which other bioactive phytochemicals are also present. Due to low levels of bioactive compounds including artemisinin and flavonoids, it is necessary to increase the level of the secondary metabolites by regulating the expression of rol genes in the plant. Methods: A hybrid variety of A. annua (Hyb1209r, Shennong)developed by the Centre for Novel Agricultural Products, University of York, UK, was selected to produce transgenics of rolB and rolC genes. Genetic transformation was carried out via Agrobacterium tumefaciens GV3101 harboring rolB and rolC genes of Agrobacterium rhizogenes cloned separately. HPLC was used for the qualitative and quantitative analysis of flavonoids and artemisinin. Furthermore, thin layer chromatography (TLC)was also used to analyze artemisinin content. Results: Comparative analysis via HPLC revealed considerable enhancement in the phytochemical content of transgenic A. annua plants as compared to the wild type plant. Transgenics of rolB gene showed an average increase of 321% in rutin, 97.2% in caffeic acid, and 218.4% in myricetin, respectively. In the case of rolC gene transgenics, an average increase of 197.5% in rutin, 76.3% in caffeic acid, and 209.3% in myricetin was observed. Transgenics of rolB and rolC genes showed a 14.3%–28.6% and 2.8%–12.7% increase in artemisinin content respectively by HPLC analysis. TLC analysis showed that an average 142.2% and 110.2% enhancement in artemisinin for rolB and rolC transgenics respectively, compared with the wild type. An enhanced production of total flavonoids (average 30.2% and 25.5% increase in rolB and rolC transgenics, respectively)and total phenolics (average 34.3% and 25.8% increase in rolB and rolC transgenics, respectively)was observed as a result of transformation. Transformed A. annua plants showed improved free radical scavenging activity (average 46.5% and 29.1% increase in rolB and rolC transgenics, respectively)and total reducing power (average 32.7% and 26.4% increase in rolB and rolC transgenics, respectively)compared with untransformed plant. Conclusion: rolB and rolC genes were effective for developing A. annua plants with an enhanced level of phytochemicals.

Effect of caffeine on anti-clotting activity of warfarin in healthy male albino rabbits

Drug-drug interactions are most commonly occurring phenomenon in clinical practice. Many physicians are afraid of being involved in an allegation of malpractices due to the occurrence of any severe interaction. These interactions not only occur between drugs but also between any kind of food, tobacco smoke, caffeine and alcohol etc. Therefore, the present study was directed to inspect the effect of caffeine on the anticoagulation activity of warfarin in healthy adult male albino rabbits. Blank blood samples were collected from each rabbit. Rabbits were given warfarin [0.5mg kg-1] orally via stomach tube and blood samples were collected in PT/INR vials at various intervals. After a washout period of 14 days, warfarin was orally administrated at same dose rate along with caffeine [5 mg kg-1 every twelve hours for three days] and same sampling schedule was repeated. Prothrombin time [PT] and the international normalized ratio [INR] of blood samples were determined to estimate changes in the anticoagulation activity of warfarin after its concurrent administration with caffeine. The PT data revealed that Rmax and AUC increased significantly [P

Ruptured second trimester ectopic tubal pregnancy

Ectopic pregnancy [EP] is one of the leading causes of maternal mortality. Its frequency in Pakistan is 1:13 pregnancy.1 Tubal ectopic often becomes symptomatic in first trimester by eroding the tubal wall and causing hemorrhage and shock. It is very rare for an ectopic to progress into second trimester and remain asymptomatic2. We are reporting a rare case of tubal ectopic pregnancy which ruptured at 16 weeks of gestation with non-viable fetus. We report a case of 27-year-old female who presented to the emergency of Dr. Zia Uddin Hospital, North Nazimabad, Karachi, with bleeding per vagina and abdominal pain with history of 4 months of amenorrhea. Ultrasound showed 16 weeks fetus in abdomen without cardiac activity and severe hemoperitoneum. She was diagnosed as a case of non-viable, ruptured, tubal ectopic pregnancy of 16-week gestational age. Diagnosis of ectopic pregnancy in first trimester can avert rupture and potential mortality and morbidity. Very few cases of second trimester ectopic pregnancy are reported. The purpose of this case report is to draw the attention for the potential of such incidence in this region and prompt management of such situation.