Elusive Toxin in Cleistanthus collinus Causing Vasoconstriction and Myocardial Depression: Detailed NMR Analyses and Biological Studies of Cleistanthoside A.

Cleistanthus collinus leaf extracts are consumed for suicidal purposes in southern India. The boiled decoction is known to be more toxic than the fresh leaf juice. Although several compounds have been isolated and their toxicity tested, controversy remains as to which compounds are responsible for the high level of toxicity of C. collinus. We report herein that cleistanthoside A is the major toxin in the boiled aqueous extract of fresh leaves and causes death in rats in small doses. The toxicity of the boiled extract prepared in the manner described can be attributed entirely to cleistanthoside A. Cleistanthin A could also be isolated from the boiled extract, albeit in trace amounts. As hypotension not responding to vasoconstrictors is the cause of death in patients who have consumed the boiled extract, effects of cleistanthoside A on the determinants of blood pressure, namely, force of cardiac contraction and vascular resistance, were tested in isolated organ experiments. Cleistanthoside A has a direct vasoconstrictor effect; however, it inhibits ventricular contractility. Therefore, the notion that the shock in C. collinus poisoning is of vascular origin must be considered carefully, and the possibility of cardiogenic shock must be studied. We present the crystal structure of cleistanthin A and show the potency of fast NMR methods (NOAH4-BSCN-NUS) in the full spectral assignment of cleistanthoside A as a real-world sample of a natural product. We also compare the results of the NOAH4-BSCN-NUS NMR experiments with conventional NMR methods.

Rat skeletal muscle-nerve preparation to teach skeletal muscle physiology.

This sourcebook update describes a variation of a previous sourcebook experiment that used isolated extensor digitorum longus muscle from mouse to teach skeletal muscle properties (Head SI, Arber MS. Adv Physiol Educ 37: 405-414, 2013). Gastrocnemius-sciatic nerve preparation in an anaesthetized rat was developed and muscle contractions were recorded in a computerized data acquisition system using an isometric force transducer. Teachers and students in physiology or biology can use this preparation to demonstrate skeletal muscle properties like simple muscle twitch, quantal summation, wave summation, superposition, incomplete tetanus, complete tetanus, treppe, fatigue, and length-tension relationship.