STUDIES ON HUMAN LENSES: II. DISTRIBUTION AND SOLUBILITY OF FLUORESCENT PIGMENTS IN CATARACTOUS AND NON-CATARACTOUS LENSES OF INDIAN ORIGIN

Fluorometric studies of cataractous and non-cataractous human lenses were carried out to study the emission characteristics and the distribution and solubility of lenticular pigments. Most of the detected fluorophores were well distributed over the cortical and nuclear portion of the lens. The decrease in solubility of proteins with aging and cataract formation is concomitant with increasing photolysis of tryptophan. However, this is likely a phenomenon independent of the photochemical transformations of the lens proteins. The number of emitting species in the diseased lenses are higher than in the normal mature lenses. A species emitting around 375 or 388 nm is of particular interest (lambda cx 330 nm) in that the emission characteristics of this fluorophore resemble kynurenic acid which has a high photosensitizing efficiency. The concentration of fluorescent pigments in the lenses of Indian origin is significantly high. The intense pigmentation could be attributed largely to the formation of photoproducts in the absence of normal endogenous antioxidant accumulation that is dependent on nutrition standard. If, indeed, any of these fluorescent pigments, because of their photosensitizing ability, are responsible for lenticular opacity, it is not the abundance of sunlight alone but also malnutrition that could account for the high incidence of cataract in India.     

STUDIES ON HUMAN LENS: I. ORIGIN AND DEVELOPMENT OF FLUORESCENT PIGMENTS

Fluorescence spectra of normal mature human lenses have been measured and at least eight species with distinct emission characteristics identified. To determine the specific photochemical and photophysical processes responsible for the origin and development of these fluorophores, emission behavior of the products generated by successive irradiation of young human lenses (3-6 y old) as well as of L-tryptophan solution have been systematically monitored. Fluorescent products that resulted from this irradiation were comparable to many of the fluorophores detected in aged lenses, indicating that light plays a major role in the development of these pigments. In addition to photogenerated species, there are other compounds in human lenses, presumably advanced glycosylated end products, with marked fluorescence properties. Several oxidation products of tryptophan including N-formylkynurenine or its derivatives, beta-carboline or its derivatives, and anthranilic acid have been identified in the mature human lens. The development of several photoproducts also was attributed to endogenous ascorbate-mediated Maillard reaction products, which undergo photoconversion by the visible light. Although some of these chromophores could act as photosensitizers, the sensitizing efficiency of many are low. Conversely, the near-UV filtering capability of these colored compounds conceivably could protect the vitreous and retina from development of any photochemical lesion.     

STUDIES ON HUMAN LENS: I. ORIGIN AND DEVELOPMENT OF FLUORESCENT PIGMENTS

Abstract
Fluorescence spectra of normal mature human lenses have been measured and at least eight species with distinct emission characteristics identified. To determine the specific photochemical and photophysical processes responsible for the origin and development of these fluorophores, emission behavior of the products generated by successive irradiation of young human lenses (3–6 y old) as well as of L-tryptophan solution have been systematically monitored. Fluorescent products that resulted from this irradiation were comparable to many of the fluorophores detected in aged lenses, indicating that light plays a major role in the development of these pigments. In addition to photogenerated species, there are other compounds in human lenses, presumably advanced glycosylated end products, with marked fluorescence properties.
Several oxidation products of tryptophan including N -formylkynurenine or its derivatives, β-carboline or its derivatives, and anthranilic acid have been identified in the mature human lens. The development of several photoproducts also was attributed to endogenous ascorbate-mediated Maillard reaction products, which undergo photoconversion by the visible light. Although some of these chromophores could act as photosensitaizers, the sensitizing efficiency of many are low. Conversely, the near-UV filtering capability of these colored compounds conceivably could protect the vitreous and retina from development of any photochemical lesion.     

MICROSPECTROPHOTOMETRIC STUDIES ON THE PIGMENTS IN VIVO OF SINGLE ALGAL CELLS-I. PIGMENTS OF CHLORELLA PYRENOIDOSA

Abstract— The pigments in vivo of single cells of Chlorella pyrenoidosa were studied by the microspectrophotometric technique. An accessory recording the first derivative of absorption was used to obtain fine resolution and enhanced accuracy. The results suggest that there are several long-wavelength components of Chl a in vivo. In addition, there seem to be four short-wave forms of Chl a. It is also likely that Chl b exists in vivo in two different forms. The existence of all these forms was demonstrated at room temperature.     

蚕丝丝素中色氨酸含量及其在丝素纤维中的径向分布研究

通过双波长分光光度法和对二甲氨基苯甲醛显色法研究桑蚕春茧内、中、外各茧层丝素中的色氨酸含量和色氨酸在丝素纤维中的径向分布状况 ,并探讨了光、热处理后蚕丝丝素中色氨酸含量的变化 .结果表明 ,外、中、内各茧层丝素的色氨酸含量不一致 ,由外至内各茧层丝素中色氨酸含量依次降低 ;色氨酸在丝素纤维的径向分布不均一 ,呈现色氨酸在易溶落的最外表面层含量最多 ,次表层相对较少 ,中间层较多 ,而内层最少的分布状态 ;光或热处理均会破坏丝素中色氨酸 ,使色氨酸含量明显下降 .     

STUDIES ON THE ORIGIN OF B CHROMOSOMES IN RYE

The morphological structure, classification and meiotic behaviour as well as the possible origin of the extra centric fragment produced after intercellular chromatin migration (cytomixis) in pollen mother cells during synizesis in rye have been studied by the squash and Giemsa·C-banding techniques. The results showed that the structures of extra centric fragments are metacentric, submetacentric, telocentric or subtelocentric, being quite different from each other. Their originations are stochastic, because any chromosome can be formed by the fragment. The meiotic behaviour of extra centric fragments is identical with that of B chromosomes. We consider that extra centric fragments are B chromosomes. The fragments originated from the fragmentation of A chromosomes at synizesis during cytomixis. Thus we consider that cytomixis could be one of the origins of B chromosomes.     

INFRARED AND RAMAN STUDIES ON SOME IMINES AND THEIR PICRATES. RELATION TO THE PROBLEM OF PROTONATION IN VISUAL PIGMENTS

Abstract— Picrates of conjugated Schiff bases have C=N stretching frequencies similar to those of the respective free bases. This raises the question if the frequency of this vibration can provide proof for the protonation of the retinylidene Schiff base occurring in visual pigments.     

ON THE ORIGIN OF LOW TEMPERATURE THERMOLUMINESCENCE IN NUCLEIC ACID BASES: EXCITATION AND EMISSION SPECTRAL STUDIES

Abstract. Thermoluminescence excitation spectra of adenine, guanine, thymine and 1,3-dimethyluracil were observed to be similar to their phosphorescence excitation spectra. Intensity dependence studies of thermoluminescence induced by ultraviolet light suggest that thermoluminescence could not be due to biphotonic ionisation of the molecule by UV. Evidence is presented which demonstrates that the fluorescence component observed in thermoluminescence of some compounds may not be due to triplet-triplet fusion. Further, phosphorescence and thermoluminescence measurements following excitation with monochromatic light at very low intensities and short durations have shown for both a linear dependence on excitation intensity. It thus appears that direct entry of the electrons from the traps into the singlet manifold is necessary for explaining the fluorescence component of thermoluminescence.     

蚕丝丝素纤维中氨基酸在丝素纤维的径向分布研究

通过显微镜观察、红外光谱、氨基酸组成分析、福林试剂分析等方法研究蚕丝丝素纤维的微结构、丝素表层和内层的结构差异 ,研究氨基酸尤其是对丝素纤维的结构和性能起重要影响的主要芳香族氨基酸———酪氨酸在丝素由表及里各层中的含量和分布 ,以期最终揭示丝素结构与蚕丝织物应用中的固有缺陷 (如泛黄、易皱、色牢度等 )的关系 .结果表明蚕丝丝素存在多层次结构 ,表层无定形区的比例较高 ,里层结晶区的比例较高 ;侧基较大的氨基酸和赖氨酸、组氨酸、胱氨酸、精氨酸、天门冬氨酸、苯丙氨酸、亮氨酸、谷氨酸、酪氨酸等在丝素的表层分布相对较多 ,而乙氨酸、丙氨酸、丝氨酸等小侧基氨基酸则在中间层至里层较多     

STUDIES ON PLANT BILE PIGMENTS. II. REGIOSELECTIVE PHOTOCHEMICAL AND ACID CATALYZED Z,E ISOMERIZATION OF DIHYDROBILINDIONE AS PHYTOCHROME MODEL

The (4 Z, 10 Z , 15 E )-2,3-dihydrobilindione 4 , along with the fully unsaturated ( E, Z, Z )-analogue 8 , has been prepared from the corresponding ( Z, Z, Z )-isomer by a variation of Falk's method (Falk et aL, 1980). The photochemical and acid-catalyzed back-reactions have been studied by UV-vis and ¹H-NMR spectroscopy.     

DISTRIBUTION AND ELIMINATION OF PHOTOFRIN II IN MICE

The distribution and elimination of [14C]PII, the radioisotopically-labeled equivalent of the mixture of porphyrins known as Photofrin II used in the photodynamic treatment of solid tumors, were determined in tumor-free and SMT-F tumor-bearing DBA/2 Ha-DD mice. Following i.p. injection, drug was absorbed from the peritoneum with a half-life of about 1 h; elimination from plasma was rapid, declining about 1.4 logs in concentration over 48 h following i.v. administration. However, some [14C]-activity was still detectable after 75 days. Normal tissues take up the drug within about 7.5 h after administration, with peak concentrations distributed as follows: liver, adrenal gland, urinary bladder greater than pancreas, kidney, spleen greater than stomach, bone, lung, heart greater than muscle much greater than brain. Only skeletal muscle, brain, and skin located contralaterally to subcutaneously implanted SMT-F tumors had peak [14C]-activities lower than tumor tissue; skin overlying SMT-F tumors showed concentrations not significantly different (P greater than 0.3) from tumor. After 75 days all tissues examined retained some fraction of [14C]-activity, ranging from 16% for kidney to 61% for spleen, of the initial peak tissue levels. The primary route of elimination of Photofrin II was through the bile-gut pathway, with greater than 59% of the administered [14C]-activity recovered in the feces, and only about 6% in the urine, over 192 h. HPLC analyses of fecal extracts showed that mostly monomeric and other low molecular weight porphyrin components of Photofrin II were eliminated. The higher molecular weight oligomeric fractions of Photofrin II were retained in liver and spleen up to 14 days after injection.     

激光诱导荧光光谱分析法研究：Ⅴ.Dy—TFA—TOPO—Tween—20的荧光特性及应用

激光诱导荧光光谱在确定与证实光谱的归属和能量传递研究以及提高检测灵敏度方面,具有很大的潜力。前人有关稀土配合物的激光诱导荧光分析虽能提高测定灵敏度,但很少涉及发光机理和能量传递方面的研究。     

STUDIES ON THE LOCALIZATION, FUNCTION, AND FORMATION OF THE CAROTENOID PIGMENTS OF A STRAIN OF MYCOBACTBRIUM MARINUM

Abstract— Aninvestigation was made of the location, function and characteristics of production of carotenoid pigments of a strain of Mycobacterium marinum , an organism which produces its pigments only when exposed to light.
It was found that light-grown cells of this organism contain only one pigment. The intracellular location of this pigment was found to be the cell envelope. The pigment was found to be capable of protecting the cells against lethal photo-oxidations mediated by the dye, toluidine blue.
These studies also revealed that in dark-grown cells exposed to light, pigment production occurred after a short lag and then remained constant during logarithmic growth. Pigment was produced by starved cells, but to a lesser degree than son-starved cells; the addition of a carbon or nitrogen source to starved cells increased pigmentation to the level in non-starved cells. Pigmentation did not occur in the absence of oxygen. Cells exposed to light at 4°C were not pigmented immediately after exposure, but became so if incubated at 30°C. Production of pigment was found to obey the Bunsen-Roscoe reciprocity law. The determination of an action spectrum for the production of pigment indicates that maximum pigment productiion occurs in the vicinity of 410 and 439 mµ, with smaller maxima in the vicinity of 502 , 528 and 560 mµ.     

FORMATION OF PHOTOPRODUCTS LETHAL FOR HUMAN CELLS IN CULTURE BY DAYLIGHT FLUORESCENT LIGHT AND BILIRUBIN LIGHT

Abstract. Irradiation of Dulbecco's modified Eagle's tissue culture medium with "Daylight,""Special Blue," or "Bilirubin" fluorescent light produces photoproducts lethal to human cells. Killing is abolished when (1) riboflavin, (2) tryptophan and tyrosine, or (3) riboflavin, tryptophan and tyrosine are deleted from medium prior to irradiation with any of the above fluorescent lamps. Toxic photoproducts are also formed when buffered salt solutions containing (a) riboflavin and tryptophan, (b) riboflavin and tyrosine, or (c) riboflavin, tryptophan and tyrosine are exposed to any of these light sources.     

CONFIGURATIONAL PHOTOISOMERIZATION OF BILIRUBIN IN VITRO–II. A COMPARATIVE STUDY OF PHOTOTHERAPY FLUORESCENT LAMPS AND LASERS

Abstract— A comparative investigation of configurational photoisomerization of bilirubin (Z.Z-BR) bound to human serum albumin (HSA) in vitro produced by fluorescent lamps currently employed in phototherapy of neonatal jaundice has been carried out by using a fast absorbance technique. Photoequilibrium Z, E-BR concentrations and rise-times are reported. Similar measurements effected with monochromatic laser lines in the blue-green spectral region are presented for comparison purposes.
Narrow-spectrum fluorescent lamps (violet, special-blue, filtered-green) produce Z,E-BR concentrations at photoequilibrium almost equal to those obtained with monochromatic lines. The photoequilibrium rise-times, however, strongly depend on the spectral bandwidth of the excitation light. Special-blue, daylight and green lamps produce Z,E-BR concentrations = 34,31, and 12%, respectively. For green lamps, the intense UV-blue-Hg lines are responsible for more than 50% of Z,E-BR concentration. Green light is found to be quite ineffective in forming Z,E isomers and very efficient in causing Z,E-BR to revert to native Z.Z-BR. Moreover, simultaneous irradiation of BR solution with special-blue and green fluorescent lamps produces almost complete inhibition of the Z→E reaction at suitable green/blue intensity ratio.
On the basis of these results a possible mechanism is presented to explain the clinical success of fluorescent green lamp phototherapy and the greater production of structural BR isomers recently reported for green than that for white lamps.     

ROLE OF HYDROXYL-BEARING AMINO ACIDS IN DIFFERENTIALLY TUNING THE ABSORPTION SPECTRA OF THE HUMAN RED AND GREEN CONE PIGMENTS

The human red and green cone pigments differ at either 15 or 16 amino acids, depending upon which polymorphic variants are compared. Seven of these amino acid differences involve the introduction or removal of a hydroxyl group. One of these differences, a substitution of alanine for senne at position 80, was found previously to produce a 5 nm blue shift. To determine the role of the remaining six hydroxyl group differences in tuning the absorption spectra of the human red and green pigments, we have studied six site-directed mutants in which single amino acids from the green pigment have been substituted for the corresponding residues in the red pigment. Blue shifts of 7 and 14 nm were observed upon substitution of phenylalanine for tyrosine at position 277 and alanine for threonine at position 285, respectively. Single substitutions at positions 65, 230, 233, and 309 produced spectral shifts of 1 nm or less. These data are in good agreement with a model based upon sequence comparisons among primate pigments and with the properties of site-directed mutants of bovine rhodopsin. Nonadditive effects observed in comparing the absorption spectra of red-green hybrid pigments remain to be explained.     

2,6—二乙酰吡啶类双希夫碱配合物的合成和研究 Ⅱ.Cu(Ⅱ)配合物的合成和研究

合成了五个2,6-二乙酰吡啶缩肼基硫代甲酸酯双希夫碱合铜(Ⅱ)配合物,用元素分析、磁化率、红外光谱及紫外-可见光谱等对它们进行了表征,研究了配合物的电化学性质和Cu_3L_2~2X_2(X=Cl~-、SCN~-)的光电子能谱。结果表明:五个Cu(Ⅱ)配合物均为三核配合物,在Cu_3L_2~2(SCN)_2中存在Cu(Ⅱ)间的自旋—交换作用。     

MMA-EA-AA无皂共聚胶乳中羧基分布的研究

用无皂乳液聚合法合成了粒径窄分布的MMA EA AA三元共聚胶乳 ,用电导滴定法测定了羧基在胶乳中的分布 ,系统研究了聚合体系中各组分对胶乳中羧基分布的影响 .结果表明随着NH4 HCO3用量的增加 ,表面羧基含量 (Sa)、表面羧基密度 (Sd)和水相羧基含量 (Fa)逐渐增大 ,包埋羧基含量 (Ea)明显降低 ;随着AA用量的增加 ,Sa、Sd 、Fa 和Ea 均逐渐增大 ,但包埋羧基百分比也呈增大趋势 ;随着引发剂用量的增加 ,Sd 和Ea 逐渐减小 ,而Fa 先增大然后趋于恒定 .     

2,6-二乙酰吡啶类双希夫碱配合物的合成和研究 Ⅱ.Cu(Ⅱ)配合物的合成和研究

合成了五个2,6-二乙酰吡啶缩肼基硫代甲酸酯双希夫碱合铜(Ⅱ)配合物,用元素分析、磁化率、红外光谱及紫外-可见光谱等对它们进行了表征,研究了配合物的电化学性质和Cu₃L₂²X₂(X=Cl^-、SCN^-)的光电子能谱。结果表明:五个Cu(Ⅱ)配合物均为三核配合物,在Cu₃L₂²(SCN)₂中存在Cu(Ⅱ)间的自旋-交换作用。