Reactions of the hydrated electron with pyrimidine nucleosides halogenated at the sugar moiety: e.s.r. and spin-trapping with 2-methyl-2-nitrosopropane

Free radicals produced by the reactions of hydrated electrons with pyrimidine nucleosides halogenated at the sugar moiety (2'-chloro-2'-deoxyuridine and 2'-chlorothymidine) were studied by e.s.r. and spin-trapping. 2-Methyl-2-nitrosopropane was used as the spin-trap. The usual spin-trapping technique was extended to frozen and deoxygenated systems to avoid contamination of the trapped radicals with side-products by spin-trapping 2-methyl-2-nitrosopropane itself. When this method was applied to 2'-chloro-2'-deoxyuridine, a free radical at the C-2' position of the sugar moiety was spin-trapped together with a free radical at the C-5 position of the base moiety. This indicates that hydrated electrons both add to the base moiety and eliminate halogen anions from the halogenated sugar moiety. In the case of 2'-chlorothymidine, however, only a free radical attributed to H-addition at the C-6 position of the thymine base was observed. No radicals produced by the reaction of hydrated electrons with the halogenated sugar could be spin-trapped.     

Quantitative Analysis of the Inactivation of Photosynthetic Oxygen Evolution and the Release of Polypeptides and Manganese in the Photosystem II Particles of Spinach Chloroplasts

Photosynthetic oxygen evolution by photosystem II particleswas inactivated by treatment with NaCl, NH₂OH or high pH. Whenthe degree of inactivation was compared with the degree of releasefrom the particles of Mn and three polypeptides having molecularmasses of 33, 24 and 18kdaltons, two types of inactivation werefound: one, brought about with 960 mM NaCl, was related to therelease of the 24 kdalton polypeptide, and the other, broughtabout with 1.5 mM NH₂OH or high pH, seemed to be related tothe release of Mn. ¹Present address: Department of Chemistry, Faculty of Science,Toho University, Miyama 2-2-1, Funabashi 274, Japan. (Received January 31, 1983; Accepted March 28, 1983)     

Fine structure of the dorsal ocellus of the worker honeybee

The three dorsal ocelli of worker honeybees have been studied by light and electron microscopy. Each ocellus has a single flattened spheroidal lens and about 800 elongated retinular cells. Retinular cells are paired and form a two-part plate-like rhabdom between their distal processes. Each rhabdomere comprises parallel microvilli projecting laterally from the apposed retinular cells. Primary receptor cell axons synapse within the ocellus with ocellar nerve fibers of two different calibers. Each ocellus has eight thick fibers ca 10 m?m in diameter and several thinner ones less than 3 m?m in diameter. Fine structural evidence suggests that retinular axons end presynaptically on both types of ocellar nerve fibers. Since all retinular cells apparently synapse repeatedly with the thick fibers this involves a convergence of about 100:1. Thick fibers always terminate postsynaptically within the ocellus while thin fibers terminate presynaptically on other thin fibers, thick fibers or retinular axons. Structural evidence for synaptic polarization indicates that retinular cells and thick fibers are afferent, thin fibers efferent. Thus complex processing of the ocellar visual input can occur before the secondary neurons of the three ocelli converge to form the single short ocellar nerve which runs to the posterior forebrain.     

The Development of the Rhabdom and the Appearance of the Electrical Response in the Insect Eye

Electron microscopic studies on the development of the rhabdom in the compound eye of the silkworm moth and pupa (Bombyx mori) were carried out in parallel with the recording of the electrical response to photic stimulation. No electrical response to photic stimulation was recorded from the pupal compound eye which had no trace of differentiation of the rhabdom. With the differentiation of development of the rhabdom in the pupal compound eye, electrical responses could be recorded, and the amplitude of such electrical responses increased with the progress of development of the rhabdom. These observations suggest that the rhabdom is probably the site of the photochemical reaction which leads to the generation of the slow retinal action potentials.     

Stain Technology: A Simplified Aluminum Stain in Paraffin Sections of Bone from Hemodialysis Patients

A new simplified method has been devised for staining aluminum and has been tested in paraffin sections of bone from 60 patients who have undergone hemodialysis. Iliac crest bone fragments were fixed in 20% phosphate-buffered formalin for less than a day and demineralized at room temperature in 10% phosphate-buffered formalin containing 5% formic acid for only 2 to 3 hr. Four-micron paraffin sections, accompanied by positive controls, were stained with Maloney's aluminum stain, the Berlin blue reaction for iron, dylon or Congo red for amyloid and von Kossa's reaction for calcium. Aluminum and iron were demonstrated particularly at the mineralizing front of bony tissues; aluminum in 52 cases, iron in 45. Dylon staining also gave positive results in 52 cases. It is important in determining whether aluminum deposition is present that the von Kossa reaction remains positive even after demineralization. This method may be more useful for demonstrating aluminum in bony tissues than the complicated and time-consuming resin-embedding method currently used.     

Striatal L-DOPA Decarboxylase Activity in Parkinson's Disease In Vivo: Implications for the Regulation of Dopamine Synthesis

Abstract: L-DOPA is a large neutral amino acid subject to transport out of, as well as into, brain tissue. Competition between dopamine synthesis and L-DOPA egress from striatum must favor L-DOPA egress if decarboxylation declines relatively more than transport in Parkinson's disease. To test this hypothesis, we injected patients with Parkinson's disease with a radidabeled analogue of L-DOPA and recorded regional brain radioactivity as a function of time by means of positron emission tomography. We simultaneously estimated the activity of the decarboxylating enzyme and the amino acid transport. In the striatum of patients, we found the L-DOPA decarboxylase activity to be reduced in the head of the caudate nucleus and the putamen. However, the rate of egress of the DOPA analogue was unaffected by the disease and thus inhibited dopamine synthesis more than predicted in the absence of L-DOPA egress.     

Regulation of DOPA Decarboxylase Activity in Brain of Living Rat

Abstract: To test the hypothesis that l -DOPA decarboxylase (DDC) is a regulated enzyme in the synthesis of dopamine (DA), we developed a model of the cerebral uptake and metabolism of [³H]DOPA. The unidirectional blood-brain clearance of [³H]DOPA ( K ^D₁) was 0.049 ml g⁻¹ min⁻¹. The relative DDC activity ( k ^D₃) was 0.26 min⁻¹ in striatum, 0.04 min⁻¹ in hypothalamus, and 0.02 min⁻¹ in hippocampus. In striatum, 3,4-[³H]dihydroxyphenylacetic acid ([³H]DOPAC) was formed from [³H]DA with a rate constant of 0.013 min⁻¹, [³H]homovanillic acid ([³H]HVA) was formed from [³H]DOPAC at a rate constant of 0.020 min⁻¹, and [³H]HVA was eliminated from brain at a rate constant of 0.037 min⁻¹. Together, these rate constants predicted the ratios of endogenous DOPAC and HVA to DA in rat striatum. Pargyline, an inhibitor of DA catabolism, substantially reduced the contrast between striatum and cortex, in comparison with the contrast seen in autoradiograms of control rats. At 30 min and at 4 h after pargyline, k ^D₃ was reduced by 50% in striatum and olfactory tubercle but was unaffected in hypothalamus, indicating that DDC activity is reduced in specific brain regions after monoamine oxidase inhibition. Thus, DDC activity may be a regulated step in the synthesis of DA.     

Phototransformation of the Far-red Light-absorbing Form of Large Pea Phytochrome by Laser Flash Excitation

Phototransformation of the far-red light absorbing form (P_FR)of large pea phytochrome to the red-light absorbing form (P_R)was examined at 2?C after a 715 nm laser flash excitation usinga custom-built multichannel transient spectra analyzer. Themaximum amount of phototransformation intermediates was producedby a pulse of about 50 mJ, which resulted in ca. 65% of P_R obtainedat the photostationary equilibrium. Some flash-induced intermediateswere assumed to return to P_FR in the dark. A difference spectrummeasured at 10 µsec after the flash showed an absorbanceincrease at 651 nm and a decrease at 724 nm. When the samplewas left in darkness after the flash light irradiation, absorbancein the red and far-red region gradually increased, but thatin the green region rapidly decreased. The decay curve of intermediatesmeasured at 554 nm could be resolved into three reaction componentshaving rate constants of 2,500, 590 and 48 sec^–1, respectively.Difference spectra also indicated that a small but significantincrease in absorbance between 370 and 380 nm and a decreasearound 415 nm took place 10–310 µsec after a flash. (Received February 13, 1982; Accepted April 21, 1982)