Acclimation to temperature and irradiance modulates PSII charge recombination.

Acclimation of wild type and the chlorina F2 mutant of barley to either high light or low temperature results in a 2- to 3-fold increase in non-photochemical quenching which occurred independently of either energy-dependent quenching (qE), xanthophyll cycle-mediated antenna quenching or state transitions. Results of in vivo thermoluminescence measurements used to address this conundrum indicated that excitation pressure regulates the temperature gap for S(2)Q(B)(-) and S(2)Q(A)(-) charge recombinations within photosystem II reaction centers. This is discussed in terms of photoprotection through non-radiative charge recombination.

Mitochondria/nuclear signaling of alternative oxidase gene expression occurs through distinct pathways involving organic acids and reactive oxygen species.

Cultured cells of tobacco (Nicotiana tabacum L. cv Petit Havana) were used to investigate signals regulating the expression of the "model" nuclear gene encoding the alternative oxidase (AOX) (AOX1), the terminal oxidase of the mitochondrial alternative respiratory pathway. Several conditions shown to induce AOX1 mRNA accumulation also result in an increase in cellular citrate concentrations, suggesting that citrate and/or other tricarboxylic acid (TCA) cycle intermediates may be important signal metabolites. In addition, mitochondrial reactive oxygen species (ROS) production has recently been shown to be a factor mediating mitochondria-to-nucleus signaling for the expression of AOX1. We found that the exogenously supplied TCA cycle organic acids citrate, malate and 2-oxoglutarate caused rapid and dramatic increases in the steady-state level of AOX1 mRNA at low, near physiological concentrations (0.1 mM). Furthermore, an increase in AOX1 induced by the addition of organic acids occurs independently of mitochondrial ROS formation. Our results demonstrate that two separate pathways for mitochondria-to-nucleus signaling of AOX1 may exist, one involving ROS and the other organic acids.

Authentic standards for the reductive-cleavage method. The positional isomers of partially methylated and acetylated or benzoylated methyl 2-(acetylmethylamino)-2-deoxy-beta-D-glucopyranoside.

Described herein is the synthesis of eight positional isomers of methylated and acetylated or benzoylated methyl 2-(acetylmethylamino)-2-deoxy-beta-D-glucopyranoside. The compounds were generated simultaneously from methyl 2-(acetylmethylamino)-2-deoxy-beta-D-glucopyranoside by sequential partial methylation and benzoylation and isolated in pure form by high-performance liquid chromatography (HPLC). The desired acetates were obtained by debenzoylation and acetylation of the pure isomers. Reported herein are the 1H NMR spectra of the benzoates and the electron-ionization mass spectra of the acetates and the tri-O-methyl derivative. Also reported for the acetates and the tri-O-methyl derivative are their linear temperature-programmed gas-liquid chromatography (GLC) retention indices on three different capillary columns.

Authentic standards for the reductive-cleavage method. The positional isomers of partially methylated and acetylated or benzoylated 1,5-anhydroribitol.

The eight positional isomers of methylated and benzoylated 1,5-anhydroribitol were generated simultaneously from 1,5-anhydroribitol by sequential partial methylation and benzoylation, and the four meso isomers and two enantiomeric pairs of isomers so-formed were isolated in chemically pure form by high-performance liquid chromatography. The corresponding acetates were obtained by debenzoylation of the pure isomers and acetylation. Reported herein are the 1H NMR spectra of the benzoates and the electron-ionization mass spectra of the acetates and the tri-O-methyl derivative. Also reported for the acetates and the tri-O-methyl derivative are their linear temperature-programmed gas-liquid chromatography retention indices on three different capillary columns.

Family practice career attrition, 1969 to 1993: a comparison of 336 civilian and military graduates of one medical school.

Instability of specialty choice and career after initial residency entry will affect health personnel projections. Military physicians may differ from civilian physicians in their specialty choice behaviors during their careers. A cohort of 336 graduates who chose family medicine residency training was identified from the graduating classes of 1969 through 1993 at a large private medical school. Current specialty identification was determined, and attrition from family medicine was computed. As of 1997, 275 graduates (82%) were still in family medicine careers, defined by American Academy of Family Physicians membership or current American Board of Family Practice certification. Of graduates who entered military programs, 22 of 77 (28%) had left family medicine careers; 39 of 259 (15%) of the graduates who entered civilian programs had left. This difference was statistically significant (p = 0.007). Family medicine career retention is lower for males in military programs compared with males in civilian programs or females in military programs.

Isolation and characterization of the cell-surface polysaccharides of Porphyromonas gingivalis ATCC 53978.

The cell-surface carbohydrates of Porphyromonas gingivalis strain ATCC 53978 were isolated and partially characterized. Three separate polysaccharides were found to be present: an extracellular polysaccharide, capsular polysaccharide and lipopolysaccharide. The capsular polysaccharide, which had peculiar, gel-like viscoelastic properties, was found to be comprised of mannuronic acid, glucuronic acid, galacturonic acid, galactose, and 2-acetamido-2-deoxy-D-glucose in relative molar ratios of 0.6:0.9:0.5:0.5:1.0, respectively. The extracellular polysaccharide was found to contain mannose, rhamnose, glucose, galactose, and 2-acetamido-2-deoxy-D-glucose in relative molar ratios of 13.5:1.4:1.0:2.0:1.0, respectively. The lipopolysaccharide was found to contain an O-antigen with a regular tetrasaccharide repeat unit comprised of 4-linked alpha-L-rhamnopyranosyl, 6-linked alpha-D-glucopyranosyl, 3-linked alpha-D-galactopyranosyl, and 4-linked 2-acetamido-2-deoxy-beta-D-glucopyranosyl residues in equimolar proportions.

Temperature/light dependent development of selective resistance to photoinhibition of photosystem I.

Exposure of winter rye leaves grown at 20 degrees C and an irradiance of either 50 or 250 micromol m(-2) s(-1) to high light stress (1600 micromol m(-2) s(-1), 4 h) at 5 degrees C resulted in photoinhibition of PSI measured in vivo as a 34% and 31% decrease in deltaA820/A820 (P700+). The same effect was registered in plants grown at 5 degrees C and 50 micromol m(-2) s(-1). This was accompanied by a parallel degradation of the PsaA/PsaB heterodimer, increase of the intersystem e- pool size as well as inhibition of PSII photochemistry measured as Fv/Fm. Surprisingly, plants acclimated to high light (800 micromol m(-2) s(-1)) or to 5 degrees C and moderate light (250 micromol m(-2) s(-1)) were fully resistant to photoinhibition of PSI and did not exhibit any measurable changes at the level of PSI heterodimer abundance and intersystem e- pool size, although PSII photochemistry was reduced to 66% and 64% respectively. Thus, we show for the first time that PSI, unlike PSII, becomes completely resistant to photoinhibition when plants are acclimated to either 20 degrees C/800 micromol m(-2) s(-1) or 5 degrees C/250 micromol m(-2) s(-1) as a response to growth at elevated excitation pressure. The role of temperature/light dependent acclimation in the induction of selective tolerance to PSI photoinactivation is discussed.

Authentic standards for the reductive-cleavage method: the positional isomers of partially methylated and acetylated or benzoylated 1,5-anhydro-L-arabinitol.

Described herein is the synthesis of the eight positional isomers of methylated and acetylated or benzoylated 1,5-anhydro-L-arabinitol. The compounds were generated simultaneously from 1,5-anhydro-L-arabinitol by sequential partial methylation and benzoylation and isolated in pure from by high-performance liquid chromatography. The desired acetates were obtained by debenzoylation and acetylation of the pure isomers. Reported herein are the 1H NMR spectra of the benzoates and the electron-ionization mass spectra of the acetates and the tri-O-methyl derivative. Also reported for the acetates and the tri-O-methyl derivative are their linear temperature-programmed gas-liquid chromatography retention indices on three different capillary columns.

Fiber-coupled high-power external-cavity semiconductor lasers for real-time Raman sensing.

High-power, external-cavity semiconductor lasers with narrow bandwidth and fiber-coupled output are designed and constructed. An output power of 540 mW is coupled out of a 100-mum multimode fiber with coupling efficiency of 72% when the laser is operated at 1.1 A. The emission linewidth is as narrow as 22 GHz, and the wavelength is tunable from 779.7 to 793.0 nm. Application of such lasers to remote real-time Raman sensing of materials is also demonstrated.

Cold Acclimation and Freezing Tolerance (A Complex Interaction of Light and Temperature).

By comparing growth under five different temperature and irradiance regimes (20[deg]C and 800, 250, and 50[mu]mol m-2 s-1 and 5[deg]C and 250 and 50 [mu]mul m-2 s-1), we have examined the effects of light, temperature, and the relative reduction state of photosystem II on plant morphology, freezing tolerance (lethal temperature at which freezing injury occurs [LT50]), transcript levels of Lhcb and two cold-stimulated genes (Wcs19 and Wcs120), and photosynthetic adjustment in winter rye (Secale cereale L. cv Musketeer). We show, for the first time to our knowledge, that in addition to adjustments in photosynthetic capacity, nonphotochemical quenching capacity and tolerance to photoinhibition, the accumulation of the cold-induced transcript Wcs19, and the compact plant morphology usually associated with cold-hardening are correlated with the relative reduction state of photosystem II rather than with growth temperature or growth irradiance per se. In contrast, the acquisition of maximal LT50, as well as Lhcb and Wcs120 mRNA accumulation, appears to be dependent on both growth temperature and growth irradiance but in an independent, additive manner. The results are discussed with respect to the possible role of the modulation of chloroplastic redox poise in photosynthetic acclimation to cold-hardening temperatures and the attainment of maximal LT50.

Reductive cleavage of permethylated polysaccharides with borane-methyl sulfide complex and butyltin trichloride.

Several per-O-methylated monosaccharides and polysaccharides were used as models in an attempt to identify more convenient reagents for accomplishing reductive cleavage of glycosidic linkages. Included in the model studies were methyl alpha- and beta-D-glucopyranoside, methyl alpha- and beta-D-mannopyranoside, and methyl alpha- and beta-D-ribofuranoside. These studies led to the identification of a new promoter, butyltin trichloride, for carrying out reductive cleavage when borane-methyl sulfide complex was used as the reducing agent. These reagents were found to accomplish the reductive cleavage of per-O-methylated amylose, cellulose, and pullulan to give only the expected derivatives of 1,5-anhydro-D-glucitol. These reagents also accomplished reductive cleavage of per-O-methylated insulin to give only the expected derivatives of 2,5-anhydro-D-mannitol and 2,5-anhydro-D-glucitol. Reductive cleavage using these reagents is easy to perform, and subsequent acetylation of the products is readily accomplished in situ.

Reductive cleavage of the positional isomers of benzoylated and methylated methyl alpha-D-mannopyranoside.

The four O-benzoyl-tri-O-methyl positional isomers, the six di-O-benzoyl-di-O-methyl positional isomers, the four tri-O-benzoyl-O-methyl positional isomers, and the tetra-O-benzoyl derivative of methyl alpha-D-mannopyranoside were synthesized, characterized, and subjected to reductive cleavage in the presence of triethylsilane and trimethylsilyl trifluoromethanesulfonate. The reactions were monitored by 1H NMR spectroscopy in order to establish the rates of the reactions and the identities of the products so formed. The tetra-O-benzoyl derivative, all tri-O-benzoyl-O-methyl positional isomers, and the 2.4-di-O-benzoyl-3.6-di-O-methyl positional isomer were fully stable to reductive-cleavage conditions, but the other di-O-benzoyl-di-O-methyl positional isomers and all O-benzoyl-tri-O-methyl positional isomers were converted to their respective 1.5-anhydro-D-mannitol derivatives via reductive cleavage of their anomeric carbon-oxygen bonds. For the mono-O-benzoyl positional isomers, the rate of reductive cleavage decreased in the order 2-O-benzoyl > 6-O-benzoyl > 3-O-benzoyl approximately 4-O-benzoyl. Moreover, all mono-O-benzoyl positional isomers were cleaved at a much faster rate than the di-O-benzoyl positional isomers. These results suggest that both anchimeric assistance and inductive effects of the ester groups contribute to the observed reaction rate. Based upon these results it is suggested that reductive cleavage of benzoylated polysaccharides might serve as a means to cleave selectively at branched residues.

Semiconductor laser dynamics subject to strong optical feedback.

An experimental investigation of a semiconductor laser subject to moderate to strong optical feedback shows that sporadic power discontinuities occur everywhere along the boundary separating feedback regimes IV and V. These low-frequency fluctuations (LFF's) do not occur just near the solitary-laser threshold as previously thought. Furthermore, although the LFF's appear as power dropouts near threshold, they manifest as power jump-ups well above threshold. Theoretical modeling shows reasonable agreement with experiments.

Structural identification of a major mitogenic lipid derived from Bacillus subtilis as a glycerophosphoglycolipid.

Lipids extracted from Bacillus subtilis using a 2:1 mixture of chloroform and methanol have been found to be very mitogenic. These lipids were fractionated on a silica column and eluted with chloroform, acetone, and 60% methanol in chloroform, and the mitogenic activity was recovered in the last fraction. Further purification of the mitogenic components was achieved by HPLC on an amino-isopropyl bonded-phase column using a linear gradient of 5-20 mM ammonium acetate in a mobile phase consisting of hexane, 2-propanol, methanol, and water (5.5:8:1.5:1). Two major and several minor mitogenic peaks were observed. One major mitogenic lipid was isolated in pure form and structurally characterized by chemical degradation analysis, NMR spectroscopy, and mass spectrometry. Mild acid hydrolysis of the lipid released glycerol phosphate and a neutral glycolipid. Saponification of the lipid released a water-soluble head group and C14-C17 branched fatty acids. Total acid hydrolysis of the head group revealed the presence of glycerol and glucose in a ratio of 1:1. Mild acid hydrolysis of the head group to remove the glycerol phosphate produced a neutral partial head group. The partial head group was methylated and then analyzed by GLC-CIMS and by the reductive-cleavage method, which revealed that it was composed of nonreducing terminal glucopyranosyl, 6-linked glucopyranosyl, and 3-linked glycerol residues in equimolar proportions. Finally, the molecular weight of the permethylated head group, obtained by fast atom bombardment mass spectrometry, was 724.3340, which is consistent with the composition of two glucose residues, one glycerol residue, and one glycerol phosphate residue. On the basis of all these results, the intact mitogenic lipid was identified as 1,2-di-O-acyl-3-O-[6-(sn-glycerol-phospho)-beta-D-glucopyranosyl-(1-->6) - beta-D-glucopyranosyl]glycerol. The purified glycolipid possessed very potent mitogenic activity in a murine splenocyte proliferation assay at a concentration of 0.01-0.1 microgram/mL.

Mechanism of anomerization of cyclohexyl 2-deoxy-3,4,6-tri-O-methyl-2-(N-methylacetamido)-alpha- and beta-D-hexopyranosides under reductive-cleavage conditions.

The fully methylated cyclohexyl glycosides of 2-acetamido-2-deoxy-alpha- and beta-D-hexopyranoses having the gluco, manno, and galacto configurations were each subjected to reductive-cleavage conditions using one of three promoters, namely trimethylsilyl trifluoromethanesulfonate, a mixture of trimethylsilyl methanesulfonate and boron trifluoride etherate, or boron trifluoride etherate alone. As expected, the fully methylated 1,2-trans-linked acetamido sugar derivatives were rapidly converted to their respective oxazolinium ions with all three promoters. Surprisingly, however, the fully methylated 1,2-cis-linked acetamido sugar derivatives were also converted to their respective oxazolinium ions, albeit at a much slower rate. In the latter case, evidence was obtained for anomerization to the 1,2-trans-linked isomers under reductive-cleavage conditions. Since the anomerization was relatively slow at room temperature in dichloromethane, a modified procedure was developed in which the reaction was carried out at 70 degrees C in 1,2-dichloroethane. Using the modified procedure, all 1,2-cis- and 1,2-trans-linked acetamido sugar derivatives were rapidly converted into their respective oxazolinium ions and subsequent quenching of the reactions with anhydrous methanol gave the respective 1,2-trans-linked methyl glycoside derivatives in quantitative yield. The modified procedure is recommended for the total reductive cleavage of polysaccharides comprised of acetamido sugar residues.

Photosystem II Excitation Pressure and Development of Resistance to Photoinhibition (II. Adjustment of Photosynthetic Capacity in Winter Wheat and Winter Rye).

Winter wheat (Triticum aestivum L. cv Monopol), spring wheat (Triticum aestivum L. cv Katepwa), and winter rye (Secale cereale L. cv Musketeer) grown at 5[deg]C and moderate irradiance (250 [mu]mol m-2 s-1) (5/250) exhibit an increased tolerance to photoinhibition at low temperature in comparison to plants grown at 20[deg]C and 250 [mu]mol m-2 s-1 (20/250). However, 5/250 plants exhibited a higher photosystem II (PSII) excitation pressure (0.32-0.63) than 20/250 plants (0.18-0.21), measured as 1 - qP, the coefficient of photochemical quenching. Plants grown at 20[deg]C and a high irradiance (800 [mu]mol m-2 s-1) (20/800) also exhibited a high PSII excitation pressure (0.32-0.48). Similarly, plants grown at 20/800 exhibited a comparable tolerance to photoinhibition relative to plants grown at 5/250. In contrast to a recent report for Chlorella vulgaris (D.P. Maxwell, S. Falk, N.P.A. Huner [1995] Plant Physiol 107: 687-694), this tolerance to photoinhibition occurs in winter rye with minimal adjustment to polypeptides of the PSII light-harvesting complex, chlorophyll a/b ratios, or xanthophyll cycle carotenoids. However, Monopol winter wheat exhibited a 2.5-fold stimulation of sucrosephosphate synthase activity upon growth at 5/250, in comparison to Katepwa spring wheat. We demonstrate that low-temperature-induced tolerance to photoinhibition is not a low-temperature-growth effect per se but, instead, reflects increased photosynthetic capacity in response to elevated PSII excitation pressure, which may be modulated by either temperature or irradiance.

Reductive cleavage of glycosides with borane complexes in the presence of boron trifluoride etherate.

Mixtures consisting of five equivalents each of borane.methyl sulfide and boron trifluoride etherate per equivalent of acetal or five equivalents of various amine-borane complexes and 10 equivalents of boron trifluoride etherate readily accomplished reductive cleavage of the glycosidic linkages of several per-O-methylated monosaccharides and polysaccharides. In all cases, the expected products were obtained in the expected molar proportions and no artifactual products were observed. Reductive-cleavage analysis using these reagents is particularly convenient because the reagents themselves are easy to handle and because subsequent acetylation of the products is accomplished in situ.

Hb Lulu Island (alpha 2 beta 2 107[G9]Gly-->Asp)-beta zero- thalassemia (codon 15; TGG-->TAG), a form of thalassemia intermedia.

Hb Lulu Island [beta 107(G9)Gly-->Asp] was discovered in an East Indian female who carried a common beta zero-thalassemia allele, i.e., codon 15, TGG-->TAG (is a stop codon) in trans. Both abnormalities were detected through sequencing of the amplified beta-globin genes and were confirmed by hybridization with 32P-labeled probes. Hb Lulu Island is mildly unstable with a borderline decrease in oxygen affinity; its instability is less severe than that of Hb Burke or beta 107(G9)Gly-->Arg. The compound heterozygosity expresses as a thalassemia intermedia with moderate anemia, a variable need for blood transfusions, Heinz body formation, and a red cell morphology which is typical for such a condition. The level of HbA2 was greatly increased (6.5-7.0%) as was the delta chain level (12% of total non-alpha) probably because of the instability of Hb Lulu Island and the decreased ability of the beta x chain to form dimers with the normal alpha chain.