The binding of host-selective toxin analogs to mitochondria from normal and ;texas' male sterile cytoplasm maize.

Tritium-labeled toxin analogs were prepared by reduction with NaB(3)H(4) of either the toxin from Helminthosporium maydis race T or a toxin component from Phyllosticta maydis. These reduced analogs had high radiochemical specific activities, high biological activities, and plant specificities identical to the native toxins. A filtration assay was developed to test the binding of these labeled analogs to isolated mitochondria. Binding was not energy dependent nor was there measurable matrical uptake. The analogs were shown to be lipophilic, a characteristic which gave rise to considerable nondisplaceable binding. Under conditions limiting nondisplaceable binding, the displaceable binding was shown to be linear with respect to toxin concentration and unsaturable. No significant differences were observed in the binding characteristics between the mitochondria from normal and male-sterile (Texas) cytoplasm maize. The findings suggest that, at physiologically relevant concentrations, these toxin analogs permeate the membranes of susceptible and resistant mitochondria alike. The lack of demonstrable specific binding does not rule out the involvement of a classical receptor site but does indicate that other kinds of molecular interactions may be involved in the mechanisms for toxicity and specificity.

Comparison of Activities of the Host-Specific Toxin of Helminthosporium maydis, Race T, and a Synthetic C(41) Analog.

It previously had been proposed that the host-selective toxin of Helminthosporium maydis race T consists of a series of unusual linear (C(35) to C(45))polyketols, of equal toxicity on a weight or molar (10(-8)-10(-9)) basis. Previous laboratory synthesis of T-toxin analogs was limited to shorter (C(15) to C(26)) versions which possessed the requisite specificity for susceptible corn (Zea mays) but were less toxic on a weight or molar (10(-6)-10(-7)) basis. In the present study, a C(41) analog with four beta-ketol units spaced by CH(2) bridges as in native toxin has been synthesized. On a weight or molar basis, it is as effective as native toxin or its purified components in stimulating NADH oxidation of mitochondria from susceptible corn, thus providing firm evidence for the correctness of the proposed structures of T-toxin. Additional support derives from the observation that C(24) and C(26) analogs with -(CH(2))(4)- and -(CH(2))(6)- bridges between ketol groups are not as effective in stimulating NADH oxidation as are C(23) and C(25) analogs with the -(CH(2))(3)- and -(CH(2))(5)- bridges of native T-toxin.It was calculated that a single molecule of the C(41) analog is at least 300 times more effective in stimulating mitochondrial oxidation than a molecule of the C(23) or C(25) analogs. This emphasizes the importance of chain length for toxicity, perhaps through perturbation of membrane functions of mitochondria and/or chloroplasts.

The backbone and side chain conformations of the cyclic tetrapeptide HC-toxin.

A study of the conformational parameters of HC-toxin and its diacetyl derivative in chloroform solution has been carried out. Two-dimensional NMR spectroscopy and the nuclear Overhauser effect have been used in order to determine connectivities (assignments and sequence) and approximate torsion angles and interproton distances. The results are consistent with a bis-gamma-turn conformation previously reported for dihydrochlamydocin. Model building based upon NMR data supports a D configuration for Ala2 and Pro4 residues.

Oxidation rates of major fatty acids in fasting neonatal pigs.

Thirty-two pigs were used to compare the oxidation rates of uniformly labeled (U-14C) palmitic (16:0), stearic (18:0), oleic (18:1) and linoleic (18:2) acids in fasting neonatal pigs. The pigs were allowed to nurse the sow for 24 to 48 h following birth. Subsequently, they were removed, an indwelling catheter was surgically placed in the external iliac vein and the pigs were fasted for 12 h to attain a postabsorptive state. The 14C fatty acids were administered as a single infusion (10 microCi) via the catheter, and recovery of the label as expired 14CO2 was determined at 45-min intervals for a 6-h period. Blood samples were taken following the infusion (15, 60, 120, 240, 360 min) to monitor activity maintained within the free fatty acid (FFA) fraction of the plasma pool. The oxidation rate of each fatty acid was corrected for the difference in dose dilution using a uniform factor based on plasma concentration of 18:1. The cumulative 6-h 14CO2 recovery rates (percentage of dose) were 19.1, 6.6, 30.1 and 13.1% for 16:0, 18:0, 18:1 and 18:2, respectively. Oleic acid was oxidized at a more (P less than .05) rapid rate than the other fatty acids. Palmitic acid and 18:2 were oxidized more rapidly than 18:0, although the difference between 18:0 and 18:2 was not significant. Plasma FFA pools differed with respect to the proportion of infused activity remaining at various times after administration. At 60 and 120 min postinfusion, the greatest (P less than .05) proportion of activity was maintained in the 18:1 pool (11.9 and 6.6%, respectively, vs 7.7 and 4.3% for 16:0, 6.9 and 3.9% for 18:2 and 3.6 and 2.2% for 18:0). Palmitic acid and 18:2 had a greater (P less than .05) level of activity in the plasma FFA pool at 60 min than did 18:0. This same pattern was observed through 2 h, but by 240 min postinfusion, the proportion of activity remaining in each of the plasma pools was similar. Rate of oxidation appeared to corrrespond with plasma concentration and proportion of activity remaining in the plasma FFA pool.

Ultrastructure of baroreceptors in the carotid sinus of the rabbit.

In the rabbit, baroreceptive intercalated and terminal receptor segments of the sinus nerve extend in the border region of adventitia and media of the carotid sinus wall. these receptors measuring 600-6,500 nm harbour a great number of mitochondria. They are either round, avoid or elongated. Round receptors are present over the whole area of the carotid sinus. Large receptors appear to be missing in the upper region, where only very elongated receptors are present, which are arranged in circular array in relation to the vascular axis. The baroreceptors are enveloped by special organelle-rich terminal cells of Schwann cell origin. The receptors have a special orientation in regard to the muscle-elastic system and to collagen. Receptors contain osmiophilic and laminated bodies, signs of degeneration and vesicles of different sizes. Many tubuli as well as glycogen granules are apparent. Some of these axoplasmatic changes appear to be related to metabolic processes (turnover) occurring in young and old animals.

Thermolysis chemical ionization of a complex polar lipid.

Vaporization of a ornithine-containing polar lipid from Thiobacillus thiooxidans has been accomplished by thermolysis in a chemical ionization source. The thermolysis has been shown to be more extensive than previously thought. It occurs in at least two steps, the first being dehydration of the ornithine to produce a substituted piperidone. This fragment undergoes a facile elimination to produce two neutral lipid components: a long chain fatty acid and piperidone-containing fatty amide. The results demonstrate the utility of chemical ionization for developing an understanding of a thermolysis process.

Sympathetic innervation of the pulmonary artery, ascending aorta, and coronar glomera of the rabbit. A fluorescence microscopic study.

Adrenergic nerve fibres were demonstrated in the connective tissue of the rabbit coronar glomera by means of the formaldehyde-induced fluorescence technique for catecholamines. This type of innervation is similar to the adrenergic nerve supply to the rabbit and cat carotid body. Adrenergic fibres terminate subendothelially and only a few can be traced to type I cells in the glomera coronaria. The sympathetic innervation of the ascending aorta is exceedingly sparse in contrast to the pulmonary trunk, while vasa vasorum of the ascending aorta exhibit a dense sympathetic innervation.

Sympathetic innervation of the carotid bifurcation in the rabbit and cat: blood vessels, carotid body and carotid sinus. A fluorescence and electron microscopic study.

Two postganglionic branches of the superior cervical ganglion enter the area of the carotid bifurcation in the rabbit and the cat. The common and external carotid arteries receive a rich adrenergic nerve supply, which can be demonstrated by fluorophores of biogenic amines appearing after formaldehyde treatment. The internal carotid artery is only sparsely innervated; however, it shows a dense sympathetic supply at the site of pressor receptors. Following removal of the superior cervical ganglion, a total loss of fluorescent adrenergic nerves occurs and degeneration of nerve endings possessing dense core vesicles is conspicuous. These nerve terminals are situated mainly subendothelially in the carotid body sinusoids; they only rarely terminate on type I cells.

[The fine structure of baroreceptor's nerve endings in the wall of carotid sinus in cats]. / Die Feinstruktur der barorezeptorischen Nervenendigungen in der Wand des Sinus caroticus der Katze

The baroreceptor endings of the sinus nerve extend into the region of the external media, to the media-adventitia border, and into the adventitia of the wall of the carotid sinus. The size of the receptors, tightly filled with mitochondria, varies between 600 and 5,700 nm in diameter. The endings are accompanied by densely structured Schwann's cells, come into contact with the muscular-elastic system or are surrounded by the collagenous material of the vessel wall. This functional junction makes the reception of the tension in the vessel wall possible. Glycogen granules could be demonstrated in some axon endings. In kittens and older cats, numerous receptors were regularly found with various degrees of structural change: homogeneous and lamellar transformation of mitochondria, lamellar aggregates, lysis of axoplasm matrix, irregularities of tubules and vesicles. These modifications may be evaluated as the morphological expression of degeneration and regeneration processes. Physiological detrition is considered to be the cause for this.

Electron microscopic studies of the pressoreceptor fields of the carotid sinus of the dog.

In the dog, pressosensitive endings of the sinus nerve extend along the border between the adventitia and media of the carotid sinus wall. The axon endings, containing a great number of mitochondria, can be divided into small (600-2,000 nm) and large (6,000-8,000 nm) end swellings. In the terminal region the pressosensitive fibers are surrounded by ramified and highly structured Schwann "terminal cells". The topographic location in relation to elastic and collagenous tissue indicates a functional connection between receptors and efferent nerve endings in the immediate surroundings has been discussed in this report. Several axon endings contain variable amounts of glycogen which is regarded as an indication for the inactive metabolic state of the ending. Axonal swellings demonstrate considerable modification in structure, such as loss of structural integrity in mitochondria, the formation of lamellar fields, vesicular irregularities and disintegration of axoplasm, all of which are considered as the morphological expression of "wearing out", degeneration and possibly regeneration.