Allergic contact conjunctivitis without eyelid involvement.
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Biomedical subjects
Publications and source records attributed to R Bassi.
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The developmental profiles of the gangliosides and those of the fatty acids and long-chain bases of the total ganglioside mixture of the brain of chicken were followed from the 10th day of incubation to the 63rd posthatching day. One O-acetylated polysialoganglioside that seems specific of the earlier embryonic stage and up to 21 alkali-stable components could be recognized by high-resolution two-dimensional TLC procedures and quantified by computer-assisted two-dimensional TLC densitometry. Besides a number of gangliosides identified by co-chromatography with reference standards, 10 were of unidentified structure, and within these 4 did not belong to the gangliotetrahexosyl series. Throughout embryonic life, the ceramide portion of gangliosides was found to contain the long-chain base species with 18 carbons. Those with 20 carbons, approximately 10% of the total, were found to be present only after hatching.
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Laser and neutron scattering experiments showed that in mixed micelles of ganglioside GM2 and GT1b, a membrane mimicking system, the segregation of gangliosides may occur spontaneously. Photolabeling experiments using nitrophenylazide containing ganglioside GM1 proved that gangliosides added to cells in culture enter the cell and bind to its membrane as components of microdomains, which specifically interact with a protein of about 30 kDa. This suggests that ganglioside segregation may be a natural phenomenon. Gangliosides when added to granule cells in culture led to increase in protein phosphorylation, the effect exerted being related to the amount of ganglioside molecules inserted stably into the cell lipid layer and an increase of 0.7% of the cell original ganglioside content promoted an increase of 57% in the incorporation of 32P into cell membrane proteins. From the above results a possible relationship between ganglioside segregation and involvement of ganglioside in enzyme activity control is suggested.
Gangliosides that contain the disialosyl residue alpha-Neu5Ac-(2--8)-alpha-Neu5Ac-(2--3)- can lactonize in the presence of traces of acid and this reaction has been studied in detail on GD1b [beta-Gal-(1--3)-beta-GalNAc-(1 --4)-[alpha-Neu5Ac-(2--8)-alpha-Neu5Ac-(2 --3)]-beta-Gal-(1--4)-beta-Glc-1--1)-Cer]. Lactonization occurs rapidly at a proton-ganglioside molar ratio of less than 1. At equilibrium, the ratio of GD1b to its lactone is 3:7. The data suggest the possibility that a proton-driven lactonization of gangliosides may occur in vivo.
Plasma ACTH and cortisol levels were studied in smokers and non smokers, (exposed or not to smoke of the environment), after passive exposure to cigarette smoking. Non smokers, usually not exposed to smoke, show a rise in both hormones, whereas smokers and non smokers commonly exposed to smoke don't show any change in ACTH and cortisol levels. These data suggest that nicotine acts as an acute stimulus on the hypophysis-adrenal axis even passively inhaled.
Plasma ACTH and cortisol levels after oral administration of nicotine (chewing gum containing nicotine 2 mg) in short and long time (10 and 45 min) were studied in smokers and non smokers. Non smokers after short time administration showed significant rise in ACTH and cortisol. No modifications were seen in the other groups of subjects. These data confirm that nicotine stimulates hypophysis-adrenal axis in non smokers and that this effect is connected with nicotine contents of cigarettes rather than other volatile substances.
In this research the effect of nicotine, (smoke of cigarette), was studied in smokers and non smokers during dexamethasone inhibitory test (1 mg h 23.00). ACTH and cortisol plasma levels, physiologically suppressed at 08.00, increased, after 30 min from smoking, only in group of non smokers. These data suggest that nicotine, in non smokers, could induce a maximum stimulus on diencephalic structures, so to overcome the inhibition of dexamethasone.
By detergent treatment of isolated photosynthetic membranes from maize chloroplasts, we have prepared two-dimensional crystals of the photosystem II complex. Two distinct crystal forms are produced by this treatment. Analysis of Fourier transforms of the crystals shows that each crystal type is formed from two inverted layers. Within the rectangular 17.8 x 26.7 nm unit cell of each layer is a tetrameric structure enclosing a two-fold symmetry axis, a result implying that the basic structural unit of photosystem II is dimeric. Tris-washing, which removes proteins associated with the oxygen-evolving apparatus from the inner surface of the photosynthetic membrane, causes a distinct change in the structure of these tetramers and reveals a dimeric core complex which may be directly associated with the photosystem II machinery.
Treatment of GD1a [alpha-Neu5Ac-(2----3)-beta-GalNAc-(1----4)-[alpha- Neu5Ac-(2----3)]-beta-Gal-(1----4)-beta-Glc-(1----1)-Cer] with dicyclohexylcarbodi-imide in anhydrous methyl sulfoxide affords 94-98% of GD1a-dilactone. The involvement of the carboxyl groups of the two sialic acid residues in the lactone rings was proved by ammoniolysis and reduction experiments, which gave ganglioside derivatives containing the amide of sialic acid and N-acetylneuraminulose, respectively. 1H-N.m.r. spectroscopy showed that the lactone rings involved position 2 of each galactose residue in the ester linkages.
The polypeptides of the barley light-harvesting protein of photosystem I (LHCI) share certain epitopes. At least two of these common epitopes are present in chlorophyll a/b-protein 1 (Chla/b-P1 = CP29), as shown by cross-reacting monoclonal antibodies (14). These antibodies were employed for immunological identification of polypeptides translated in vitro in an mRNA-dependent cell-free rabbit reticulocyte lysate. The monoclonal antibody CMpLHCI:2 precipitated only one polypeptide of molecular weight 28 kD from in vitro translates primed with polyA+ RNA. No 28 kD precipitation band was found, if this antibody was mixed with a PSI-200 preparation before it was added to the translate. The translational capacity of the LHCI transcripts isolated from 12 hours greened barley was much higher than those isolated from 6 hours greened barley. Transcripts for LHCI polypeptides were also found among the polyA+ RNA of the mutant viridis-k23, which is devoid of LHCI polypeptides in its thylakoid membranes. The monoclonal antibody CMpCh1a/b-P1:1 precipitated a polypeptide of molecular weight 31 kD from in vitro translates primed with polyA+ RNA. Thus, the cross-reactivity the two antibodies show with the mature proteins is not found when the antibodies are reacted with the precursor proteins.
The chlorophyll-protein complexes of purified maize photosystem II membranes were separated by a new mild gel electrophoresis system under conditions which maintained all of the major chlorophyll a/b-protein complex (LHCII) in the oligomeric form. This enabled the resolution of three chlorophyll a/b-proteins in the 26-31-kDa region which are normally obscured by monomeric LHCII. All chlorophyll a/b-proteins had unique polypeptide compositions and characteristic spectral properties. One of them (CP26) has not previously been described, and another (CP24) appeared to be identical to the connecting antenna of photosystem I (LHCI-680). Both CP24 and CP29 from maize had at least one epitope in common with the light-harvesting antennae of photosystem I, as shown by cross-reactivity with a monoclonal antibody raised against LHCI from barley thylakoids. A complex designated Chla.P2, which was capable of electron transport from diphenylcarbazide to 2,6-dichlorophenolindophenol, was isolated by nondenaturing gel electrophoresis. It lacked CP43, which therefore can be excluded as an essential component of the photosystem II reaction center core. Fractionation of octyl glucoside-solubilized photosystem II membranes in the presence and absence of Mg2+ enabled the isolation of the Chla . P2 complex and revealed the existence of a light-harvesting complex consisting of CP29, CP26, and CP24. This complex and the major light-harvesting system (LHCII) are postulated to transfer excitation energy independently to the photosystem II reaction center via CP43.
Photosystem I (PSI) preparations with a chlorophyll a/b ratio of 6.0 were isolated from barley thylakoids using two different methods. The high-molecular-mass complex (CP1a) which is resolved by non-denaturing gel electrophoresis had the same properties as a PSI preparation (PSI-200) isolated by Triton X-100 solubilisation of thylakoids followed by sucrose gradient ultracentrifugation. This material had a chlorophyll:P700 ratio of 208:1 and was composed of three different chlorophyll-protein complexes which could be separated from each other by solubilising the PSI preparation in dodecyl maltoside followed by sucrose gradient ultracentrifugation. Approximately half of the chlorophyll, including all the chlorophyll b, was located in two antenna complexes designated LHCI-680 and LHCI-730, which were identified by their characteristic low-temperature fluorescence emission spectra. The rest of the chlorophyll a was associated with the PSI reaction centre, P700 Chla-P1, which fluoresced at 720 nm. Each chlorophyll-protein complex had a unique polypeptide composition and characteristic circular dichroic and absorption spectra. The use of dodecyl maltoside instead of dodecyl sulphate resulted in a less denatured form of LHCI-680, which fluoresced at 690 nm at 77 K. One of the sucrose gradient fractions contained a complex consisting of only LHCI-730 and P700 Chla-P1 which fluoresced at 731 nm, indicating that LHCI-730 is structurally associated with P700 Chla-P1 and quenches its fluorescence. Approximately three-quarters of the light-harvesting antenna chlorophyll was in LHCI-730, but only about one-quarter of the normal complement of LHCI-730 was required to quench the reaction centre. By reducing the amount of Triton relative to the chlorophyll concentration, a PSI preparation (chlorophyll a/b ratio of 3.5) with a chlorophyll:P700 ratio of 300:1 was isolated. It contained no photosystem II, but a significant amount of LHCII which was functionally connected to the PSI reaction centre. Reconstitution studies demonstrated that excitation energy transfer from LHCII to PSI requires the presence of LHCI-680, and we propose that, in PSI, the following linear excitation energy transfer sequence occurs: LHCII----LHCI-680----LHCI-730----P700 Chla-P1.
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Thylakoids from enzymatically separated bundle sheath and mesophyll tissue chloroplasts were examined for their chlorophyll-proteins by tube sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS-PAGE). Differences were found in distribution of chlorophyll among peaks. The chlorophyll-protein a peak (CPa), considered to be the photosystem II (PSII) reaction centre by many authors, was seen to be absent in bundle sheath thylakoid samples. The slab SDS-PAGE revealed the absence of the polypeptides present in PSII preparations of chloroplast subfractions having only PSII activity. This finding confirms Anderson's hypothesis of the structure of grana and stroma thylakoids.