Photoinhibition and Its Recovery in Two Strains of the Cyanobacterium Spirulina platensis

Two strains of Spirulina platensis, marked Sp-G and Sp-RB, werestudied for their response to high photon flux densities (PFD).Sp-RB, a gas vacuolated strain, appeared more sensitive to thehigh PFD treatment as compared with Sp-G, a non-vacuolated strain.The loss of the photosynthetic activity due to the photoinhibitorytreatment was obtained at the level of whole cells as well asthe membrane level. Sp-RB was more sensitive than Sp-G at bothlevels. Experiments using chloramphenicol during the photoinhibitionprocess, and others in which the fate of radio-active labeledthylakoid proteins was followed, indicated that the differencebetween the strains lies in the rate of loss of the Dl polypeptidewith an electrophoretic mobility of 32–34 kDa. Both strainsrecovered from the photoinhibition when placed under low PFD.The recovery process started immediately after PFD was reduced,without any observed lag period, and was sensitive to chloramphenicol.Light was required for full recovery of activity. The rate ofrecovery of the two strains studied was very similar. ¹Contribution no. 29 of the Micro-Algal Biotechnology Lab. (Received January 11, 1988; Accepted March 31, 1988)     

Bone remodelling induced by physical stress is prostaglandin E2 mediated

In vitro cultured bone cells were found to be responsive to hormones and physical forces. A simple device has been developed which enables the direct application of physical forces to tissue culture dishes to which cells are firmly attached. The physical forces created a deformation of the dish. It was found that prostaglandin E₂ synthesis underwent a rapid increase, reaching a maximum after 20 min and then declined. Concurrent with the increase in prostaglandin E₂ was an increase in cyclic AMP production, having a maximum around 15 min. The increase in cyclic AMP was blocked by indomethacin, the prostaglandin E₂ synthesis inhibitor, indicating the dependence of cyclic AMP production on the de novo synthesis of prostaglandin E₂. Prostaglandin E₂ added to cells mimicked the effect of physical forces on the production of cyclic AMP. The increase in cyclic AMP resulted from an early rise in adenyl cyclase activity (within 5 min) and a later (10 min) increase in phosphodiesterase activity. The same physical forces also stimulatedthe incorporation of thymidine into DNA after 24 h. On addition of prostaglandin E₂ the increase in DNA synthesis was also mimicked. Pretreatment of the cells with indomethacin abolished the effect of physical forces on DNA synthesis.The results suggest a stimulus receptor mechanism for physical forces which, like hormonal effectors, are mediated by prostaglandins and stimulate cyclic AMP and DNA synthesis.We believe that physical forces stimulate bone remodelling through such a stimulus receptor system, mediated by prostaglandins.     

Neuronal Shot Noise and Brownian 1/f2 Behavior in the Local Field Potential

We demonstrate that human electrophysiological recordings of the local field potential (LFP) from intracranial electrodes, acquired from a variety of cerebral regions, show a ubiquitous 1/f² scaling within the power spectrum. We develop a quantitative model that treats the generation of these fields in an analogous way to that of electronic shot noise, and use this model to specifically address the cause of this 1/f² Brownian noise. The model gives way to two analytically tractable solutions, both displaying Brownian noise: 1) uncorrelated cells that display sharp initial activity, whose extracellular fields slowly decay in time and 2) rapidly firing, temporally correlated cells that generate UP-DOWN states.     

Inactivation of photosynthetic electron flow during desiccation of desert biological sand crusts and Microcoleus sp.-enriched isolates.

Filamentous cyanobacteria, the main primary producers in biological sand crusts, survive harsh environmental conditions including diurnal desiccation/rehydration cycles. Here we describe the inactivation of photosystem II during dehydration of native crusts (NC) and Microcoleus sp. isolates grown on nitrocellulose filters (NCF). The morphology of NCF cells, visualized by scanning-transmission and atomic-force microscopy, disclosed long bacterial filaments encapsulated in extracellular polysaccharides (EPS) tubes consisting of parallel fibrils (100-400 nm wide and 50-100 nm high) oriented mostly perpendicular to the tube length. Presence of empty EPS tubes indicated a gliding capability of the cells. Desiccation of NC resulted in a rapid decline of F(o) and complete loss of F(v). These changes were accompanied by a decrease of 77 K PSII fluorescence emission relative to that of PSI, when excited at 430 nm, and a significant decrease of energy transfer from phycobilisomes to PSII. Lowering the turgor pressure through the addition of 1.5 M trehalose to natural crusts, reduced F(v)/F(m) by over 50% and was accompanied by a decrease of 77 K PSI fluorescence induced by chlorophyll excitation. Excitation of phycobilisomes resulted in a downshift of the PSI emission wavelength by 8 nm, indicative of reduced energy transfer from LHCI to the core PSI. Decline of F(v)/F(m) in trehalose-incubated NCF cells did not induce significant changes in 77 K fluorescence emission. These results suggest that alterations in energy transfer from antennae to reaction centers may be part of the survival strategy of Microcoleus.     

Aging bone score and climatic factors

Hand radiograms for osseographic assessment of bone aging status were taken from more than 7,500 individuals residing in 31 different localities and belonging to 20 ethnic groups. Multiple regression analysis was used to evaluate possible associations between bone aging parameters and several climatic factors, to wit: hours of daylight in January and July, average monthly humidity and partial vapor pressure in January and July, and one climatic index pertaining to comfort conditions in life, namely, the Bioclimatic Index of Severity of Climatic Regime. Multiple regression analysis clearly pointed to significant correlations between climatic characteristics and indices defining the relative rate of bone aging in humans; it also evinced an independent contribution of July's humidity and January's mean temperature to earliest age at which first signs of bone aging can be found. In sum, there are grounds for concluding that temperature and humidity are key factors in triggering initial bone changes in individuals within the human populations prone to environmental effects. The combination of humidity and temperature with other factors which reflect the sharpness of the interseasonal differences in climatic conditions predispose the populations to early onset of bone changes. Am J Phys Anthropol 106:349–359, 1998. © 1998 Wiley-Liss, Inc.     

Tools for the study of protein quality control systems: use of truncated homoserine trans-succinylase as a model substrate for ATP-dependent proteolysis in Escherichia coli

Protein quality control, mediated by chaperones and ATP-dependent proteases, is essential for maintaining balanced growth and for regulating critical processes. To study these systems it is necessary to have model substrate proteins. However, most cellular proteins are stable and the few unstable proteins are usually regulatory and present in low concentrations, making them unsuitable for studies, especially in vivo. We present HTS(Delta1-6), a truncated homoserine trans-succinylase (HTS) which is unstable, can be expressed at high levels and has an enzymatic, measurable, activity. This protein can serve as a good model substrate for Escherichia coli ATP-dependent proteolysis.     

Analysis in vitro of the enzyme CRTISO establishes a poly-cis-carotenoid biosynthesis pathway in plants

Most enzymes in the central pathway of carotenoid biosynthesis in plants have been identified and studied at the molecular level. However, the specificity and role of cis-trans-isomerization of carotenoids, which occurs in vivo during carotene biosynthesis, remained unresolved. We have previously cloned from tomato (Solanum lycopersicum) the CrtISO gene, which encodes a carotene cis-trans-isomerase. To study the biochemical properties of the enzyme, we developed an enzymatic in vitro assay in which a purified tomato CRTISO polypeptide overexpressed in Escherichia coli cells is active in the presence of an E. coli lysate that includes membranes. We show that CRTISO is an authentic carotene isomerase. Its catalytic activity of cis-to-trans isomerization requires redox-active components, suggesting that isomerization is achieved by a reversible redox reaction acting at specific double bonds. Our data demonstrate that CRTISO isomerizes adjacent cis-double bonds at C7 and C9 pairwise into the trans-configuration, but is incapable of isomerizing single cis-double bonds at C9 and C9'. We conclude that CRTISO functions in the carotenoid biosynthesis pathway in parallel with zeta-carotene desaturation, by converting 7,9,9'-tri-cis-neurosporene to 9'-cis-neurosporene and 7'9'-di-cis-lycopene into all-trans-lycopene. These results establish that in plants carotene desaturation to lycopene proceeds via cis-carotene intermediates.     

Cardiac contractility modulation by electric currents applied during the refractory period

Inotropic effects of electric currents applied during the refractory period have been reported in cardiac muscle in vitro using voltage-clamp techniques. We investigated how electric currents modulate cardiac contractility in normal canine hearts in vivo. Six dogs were instrumented to measure regional segment length, ventricular volume (sonomicrometry), and ventricular pressure. Cardiac contractility modulating (CCM) electric currents (biphasic square pulses, amplitude +/-20 mA, total duration 30 ms) were delivered during the refractory period between pairs of electrodes placed on anterior and posterior walls. CCM significantly increased index of global contractility (E(es)) from 5.9 +/- 2.9 to 8.3 +/- 4.6 mmHg/ml with anterior CCM, from 5.3 +/- 1.8 to 8.9 +/- 4.0 mmHg/ml with posterior CCM, and from 6.1 +/- 2.6 to 11.0 +/- 7.0 mmHg/ml with combined CCM (P < 0.01, no significant change in volume axis intercept). End-systolic pressure-segment length relations showed contractility enhancement near CCM delivery sites, but not remotely. Relaxation was not influenced. CCM increased mean aortic pressure, but did not change peripheral resistance. Locally applied electrical currents enhanced global cardiac contractility via regional changes in myocardial contractility without impairing relaxation in situ.     

Fenfluramine-induced serotonergic neurotoxicity in mice: lack of neuroprotection by inhibition/ablation of nNOS

Previous studies have implicated a role for nitric oxide (NO) and peroxynitrite in methamphetamine-induced dopaminergic neurotoxicity. The present study was undertaken to investigate whether NO is involved in serotonergic neurotoxicity caused by fenfluramine. In the first experiment, the effect of the neuronal nitric oxide synthase (nNOS) inhibitor 7-nitroindazole (7-NI; 25 mg/kg x 4) on fenfluramine (25 mg/kg x 4)-induced serotonergic neurotoxicity in Swiss Webster mice was investigated. In the second experiment, the effect of fenfluramine (25 mg/kg x 4) on nNOS (-/-) and wild-type (WT) mice was investigated. Fenfluramine induced hypothermia in all three mouse strains, and 7-NI had no thermoregulatory effect. Selective depletion of 5-HT and 5-HT transporter binding sites in the striatum, frontal cortex and hippocampus in all three mouse strains was observed, with no evidence of dopaminergic neurotoxicity. In the first experiment, 7-NI did not attenuate serotonergic neurotoxicity in Swiss Webster mice. In the second experiment, nNOS(-/-) and WT mice were equally sensitive to serotonergic neurotoxicity. These findings suggest that NO and peroxynitrite do not mediate fenfluramine-induced serotonergic neurotoxicity, and that NO is a selective mediator of amphetamines-induced dopaminergic neurotoxicity.