Induction of apoptosis by the ginsenoside Rh2 by internalization of lipid rafts and caveolae and inactivation of Akt

Background and purpose:

Lipid rafts and caveolae are membrane microdomains with important roles in cell survival signalling involving the Akt pathway. Cholesterol is important for the structure and function of these microdomains. The ginsenoside Rh2 exhibits anti-tumour activity. Because Rh2 is structurally similar to cholesterol, we investigated the possibility that Rh2 exerted its anti-tumour effect by modulating rafts and caveolae.

Experimental approach:

A431 cells (human epidermoid carcinoma cell line) were treated with Rh2 and the effects on cell apoptosis, raft localization and Akt activation measured. We also examined the effects of over-expression of Akt and active-Akt on Rh2-induced cell death.

Key results:

Rh2 induced apoptosis concentration- and time-dependently. Rh2 reduced the levels of rafts and caveolae in the plasma membrane and increased their internalization. Furthermore, Akt activity was decreased and consequently, Akt-dependent phosphorylation of Bad, a pro-survival protein, was decreased whereas the pro-apoptotic proteins, Bim and Bax, were increased upon Rh2 treatment. Unlike microdomain internalization induce by cholesterol depletion, Rh2-mediated internalization of rafts and caveolae was not reversed by cholesterol addition. Also, cholesterol addition did not restore Akt activation or rescue cells from Rh2-induced cell death. Rh2-induced cell death was attenuated in MDA-MB-231 cells over-expressing either wild-type or dominant-active Akt.

Conclusions and implications:

Rh2 induced internalization of rafts and caveolae, leading to Akt inactivation, and ultimately apoptosis. Because elevated levels of membrane rafts and caveolae, and Akt activation have been correlated with cancer development, internalization of these microdomains by Rh2 could potentially be used as an anti-cancer therapy.     

In-Vitro Assessment of the Acaricidal Properties of Artemisia annua and Zataria multiflora Essential Oils to Control Cattle Ticks

Background

The aim of this study was to investigate the ‘acaricidal effect’ of Zataria multiflora and Artemisia annua essential oils on Rhipicephalus (Boophilus) annulatus.

Methods

This study was carried out in 2009 in the Laboratory of Parasitology of the Faculty of Veterinary Medicine of Shahrekord University, west central Iran. Six dilutions (5, 10, 20, 40, 60 and 80 µL/cm³) of both essential oils were used against engorged female R. (Boophilus) annulatus ticks using an in vitro immersion method. The mortality rates for each treatment were recorded 6, 15 and 24 hours post inoculation (hpi). Mortality rate was analyzed using Repeated Measures Analysis of Variance, and comparison of means was carried out using General Linear Models Procedure.

Results

The mortality rate caused by different dilutions of Z. multiflora essential oil ranged from 26.6% (using 10 µL/cm³) to 100% (using 40 µL/cm³) and for A. annua essential oil it was 33.2 to 100% (using 20 and 80 µL/cm³, respectively) by the end of the experiment (36 hpi). No mortality was recorded for the non-treated control group or for dilutions less than 5 and 10 µL/cm³ using Zataria and Artemisia essential oils, respectively. For Z. multiflora mortality peaked at 15 hpi for all concentrations other than 20 µL/cm³ and took 24 h to achieve its maximum effect while for A. annua the two highest concentrations needed 24 hpi to reach their full effect. In addition, essential oils applied at more than 20 and 60 µL/cm³ caused 100% egg-laying failure in engorged female ticks by Zataria and Artemisia, respectively while no failure was observed for the non-treated control group. The mortality rate in both botanical acaricides was dose-dependent.

Conclusion

Both these medicinal plants have high potential acaricidal effects on the engorged stage of R. (Boophilus) annulatus in vitro.     

Possible effect of secretor locus on plasma concentration of factor VIII and von Willebrand factor

A significant fraction (30%) of the genetically determined variance in plasma concentration of the von Willebrand factor antigen (vWf:Ag) has been shown to be related to ABH determinants. Individuals with blood group O, who have the highest amounts of blood group H substance, have the lowest concentration of vWf:Ag. The Lewis substances, Le(a) and Le(b), are biochemically closely related to the ABH substances as both can be produced from the same precursor substance. We studied the effect of the presence of the Lewis antigens on the plasma concentration of vWf:Ag and factor VIII antigen (VIII:Ag) in 323 individuals of different ABO groups from a series of twins and in 58 blood donors of blood group O. Among persons belonging to blood group O, those with the Le(a) antigen had a higher concentration of both vWf:Ag and VIII:Ag than individuals lacking Le(a). Le(a+b-) people are nonsecretors and Le(a-b+) people are secretors of ABH substance. Thus, the lowest concentration of vWf:Ag and VIII:Ag was found in group O secretors. The effect is most likely due to an effect of the secretor locus. This finding may be of importance for the detection of carriers of hemophilia A and for the diagnosis of type I von Willebrand disease.     

Abnormal liver function tests associated with severe rhabdomyolysis

Rhabdomyolysis is a syndrome of skeletal muscle injury with release of cellular constituents such as potassium,phosphate,urate and intracellular proteins such as myoglobin into the circulation,which may cause complications including acute kidney injury,electrolyte disturbance and cardiac instability.Abnormal liver function tests are frequently observed in cases of severe rhabdomyolysis.Typically,there is an increase in serum aminotransferases,namely aspartate aminotransferase and alanine aminotransferase.This raises the question of liver injury and often triggers a pathway of investigation which may lead to a liver biopsy.However,muscle can also be a source of the increased aminotransferase activity.This review discusses the dilemma of finding abnormal liver function tests in the setting of muscle injury and the potential implications of such an association.It delves into some of the clinical and experimental evidence for correlating muscle injury to raised aminotransferases,and discusses pathophysiological mechanisms such as oxidative stress which may cause actual liver injury.Serum aminotransferases lack tissue specificity to allow clinicians to distinguish primary liver injury from muscle injury.This review also explores potential approaches to improve the accuracy of our diagnostic tools,so that excessive or unnecessary liver investigations can be avoided.