Ultraviolet photochemistry of buta-1,3- and buta-1,2-dienes: laser spectroscopic absolute hydrogen atom quantum yield and translational energy distribution measurements.

Using pulsed H-atom Lyman-alpha laser-induced fluorescence spectroscopy along with a photolytic calibration approach, absolute H-atom product quantum yields of phi(H-b13d) = (0.32+/-0.04) and phi(H-b12d) = (0.36+/-0.04) were measured under collision-free conditions for the 193 nm gas-phase laser flash photolysis of buta-1,3- and buta-1,2-diene at room temperature, which demonstrate that nascent H-atom formation is of comparable importance for both parent molecules. Comparison of the available energy fraction, f(T-b13d) = (0.22+/-0.03) and f(T-b12d) = (0.13+/-0.01), released as H+C(4)H(5) product translational energy with results of impulsive and statistical energy partitioning modeling calculations indicates that for both, buta-1,3- and buta-1,2-diene, H-atom formation is preceded by internal conversion to the respective electronic ground state (S(0)) potential energy surfaces. In addition, values of sigma(b-1,3-d-L alpha) = (3.5+/-0.2)x10(-17) cm(2) and sigma(b-1,2-d-L alpha) = (4.4+/-0.2)x10(-17) cm(2) for the previously unknown Lyman-alpha (121.6 nm) radiation photoabsorption cross sections of buta-1,3- and buta-1,2-diene in the gas-phase were determined.

Sperm motion characteristics of GarolexMalpura sheep evolved in a semi-arid tropical environment through introgression of FecB gene.

The arid and semi-arid tropical climates of India are endowed with vast diversity of non-prolific sheep breeds. The GarolexMalpura sheep has been evolved in a semi-arid tropical environment by introgression of FecB gene via artificial insemination of Malpura ewes using diluted semen of prolific microsheep Garole and subsequently multiplied by inter se mating among GarolexMalpura halfbreds. The aim of the present study was to identify FecB mutation in sexually mature GarolexMalpura rams by forced RFLP-PCR of BMPR-1B gene and evaluate: (i) semen production and sperm motion characteristics of GM rams and (ii) influence of age and FecB genotype on their semen attributes. Semen was collected during autumn season from 12 donor rams by artificial vagina on 8 occasions at weekly interval. The overall means of traits which did not differed significantly with age or FecB genotyping were volume (0.72 ml), mass motility (4.44), sperm concentration (2721.56 x 10(6)ml(-1)), curvilinear velocity (134.51 microm/s), motility (81.3%), amplitude of lateral head displacement (6.24 microm), beat frequency (44.43 Hz), sperm head elongation (48.9%) and sperm head area (10.01 microm(2)). The FecB genotyping had a significant effect (P<0.05) on percent linearity and rapid motile sperms, which did not vary significantly with age. Although sperm concentration was higher in FecB(BB) and FecB(B+), compared to FecB(++) genotypes but the effect was non-significant. The age and FecB genotyping had significant effects (P<0.05) on straightness, average path velocity, straight-line velocity and percentage of medium or slow motile sperms. It is concluded that GarolexMalpura rams with introgressed FecB gene are capable of producing good quality semen in a semi-arid tropical climate.

Production of prolific microsheep by embryo transfer into large non-prolific sheep.

The Garole is a prolific breed of microsheep that possesses the FecB gene, which increases ovulation rate. The purpose of this study was to compare embryo production by multiple ovulation in seven Garole ewes with that in seven normal size, non-prolific Malpura ewes, and assess the influence of the large body size of Awassi crossbred recipient ewes on the birth-weight of Garole lambs. Oestrus was synchronised with two intramuscular injections of 7.5 mg prostaglandin F(2alpha) administered 10 days apart. The donor ewes were superovulated by the use of pregnant mare serum gonadotrophin and follicle-stimulating hormone. The onset and duration of oestrus were similar in both breeds. The Garole donors had higher total mean (se) ovarian responses (15.6 [3.6] v 9.1 [2.3]), ovulation rate (13.6 [3.1] v 8.4 [2.2]) and produced more transferable embryos (6.0 [3.5] v 4.0 [0.9]) than the Malpura donors, but the differences were not statistically significant. The Garole lambs produced by embryo transfer were on average 57.8 per cent heavier at birth than contemporary Garole lambs produced by natural mating.

EPR investigations on radiation induced chemical transformations in Pd(ClO4)2/i-PrOH/HClO4 system from 77 to 300 K.

The EPR studies have been carried out on gamma irradiated samples of Pd(ClO4)2 in 9 M/4M HClO4 in the presence and absence of isopropanol [i-PrOH (5%, v/v)], in the temperature range of 77-300 K. The EPR studies revealed the formation of Pd+ complex, Pd3+ complex and several radicals such as H., OH., ClO3., ClO4. and O3- on radiolysis of these systems. Reduction of Pd2+ to Pd clusters could not be observed in the absence of i-PrOH and warming of the irradiated sample finally resulted in recovery of Pd2+ complex. However, in the presence of 5% i-PrOH, strongly oxidizing radicals like OH., ClO3. and ClO4. are suppressed and (CH3)2C.OH radical was produced along with reducing species like H.. EPR spectra in these strongly reducing conditions showed formation of Pd+ complex, which on heating above 230 K, disproportionated into Pd0 and Pd2+ complex and finally lead to formation of Pd metal aggregates. Simultaneously, pulse radiolysis technique has been employed as an alternative method to verify the EPR observations.

Photophysical properties of 2-amino-9,10-anthraquinone: evidence for structural changes in the molecule with solvent polarity.

The photophysical properties of 2-amino-9,10-anthraquinone (2AAQ) have been investigated in different solvents and solvent mixtures and correlated with the Lippert-Mataga solvent polarity parameter, Deltaf. In the low solvent polarity region with Deltaf < ca. 0.1, the dye shows unusually high fluorescence quantum yields (Phif) and lifetimes (tauf) in comparison to those in other solvents of medium to high polarities. Similarly, the radiative rate constants (kf) are relatively lower and the non-radiative rate constants (knr) are relatively higher in the low polarity solvents in comparison to those in the medium to high polarity solvents. The current results have been rationalized assuming that the dye adopts different structural forms below and above the Deltaf value of approximately 0.1. It is inferred that in the low solvent polarity region the dye exists in a non-planar structure, with its 2-NH2 plane away from that of the 9,10-anthraquinone moiety in the ground state. In solvents of medium to high polarities, the dye exists in a polar intramolecular charge transfer (ICT) structure, where the amino lone pair of the 2-NH2 group is in strong resonance with the anthraquinone pi-cloud in the ground state. In all the solvents, however the dye is inferred to exist in the ICT structure in its excited (S1) state. Supportive evidence for the above hypothesis has been obtained from the solvent polarity effect on the Stokes' shifts for the dye. Quantum chemical studies on the structures of 2AAQ dye in the gas phase also give qualitative support for the inferences drawn from the photophysical properties of the dye in different solvents.

Evaluation of gross anatomical features of cervix of tropical sheep using cervical silicone moulds.

The lambing rate obtained following cervical artificial insemination (AI) with frozen semen in sheep is low mainly due to the inability of frozen-thawed sperm to traverse the tortuous nature of the cervical canal. Although acceptable fertility has been attained by circumventing the cervical barrier through laparoscope aided intrauterine AI, the emphasis is currently given on the development of alternate non-invasive transcervical AI procedures. The complex anatomy of the cervix does not facilitate easy transcervical passage for an insemination catheter. The aim of the present study was: (i) to examine the gross anatomy of the cervix in slaughtered ewe lambs and adult ewes of the native Malpura and Kheri breeds raised under semi-arid tropical environment; and (ii) to cast silicone moulds of the reproductive tracts for measuring the dimensions of the cervix. Eighty reproductive tracts were excised immediately from carcass of Malpura and Kheri ewes and the external os of each one was classified depending on their appearance as duckbill, spiral, rosette or flap. The cervical canal of each tract was filled with a silicone sealant for casting the mould. Fifty complete silicone moulds were obtained representing 25 from ewe lambs and 25 from adult ewes. The mean lengths of the cervical mould of ewe lambs and adult ewes were 3.8+/-0.12 and 5.3+/-0.15 cm, respectively. The average number of funnel shaped folds in the cervical mould of ewe lambs and adult ewes were 3.2+/-0.19 and 3.4+/-0.22. However, the second and third-folds from the os were observed to be accentric in both ewe lambs and adult ewes. The information generated in this study would be useful for increasing the success rate of penetration in ewes exhibiting estrus in order to improve the lambing rate of tropical ewes following transcervical AI.

Effect of post-thaw incubation on sperm kinematics and acrosomal integrity of ram spermatozoa cryopreserved in medium-sized French straws.

The objectives were to assess the effect of post-thaw in vitro incubation on motion characteristics and acrosomal integrity of ram spermatozoa of native Malpura and Bharat Merino breeds maintained under a semi-arid tropical environment. Good quality semen samples of both breeds were diluted, packaged in medium-sized straws, and frozen under controlled conditions. Straws were thawed at 60 degrees C for 10s and thawed samples were incubated at 37 degrees C for 4h. Post-thaw motion characteristics and acrosomal integrity of incubated spermatozoa were assessed (by computer-aided semen analysis and Giemsa staining, respectively) just prior to incubation and at hourly intervals thereafter. There was a significant effect of incubation time on motility characteristics and the proportion of spermatozoa with normal acrosomes; 81.4% (arcsin transformed value, 65.2) of spermatozoa were motile at the start of incubation, with 47.9% (arcsin transformed value, 44.4) motile after 4h. At the corresponding times, there were normal acrosomes in 65.8 (arcsin transformed value, 54.8) and 55.7% (arcsin transformed value, 48.9) of spermatozoa, respectively. The percentage straightness of spermatozoa varied during incubation (P < 0.01). However, there was no significant change in percentage linearity, curvilinear velocity, average path velocity, straight line velocity, lateral head displacement, and beat cross frequency of spermatozoa during incubation. There were no breed variations in any motility parameters during incubation, except percentage straightness (P < 0.05), lateral head displacement (P < 0.05) and beat cross frequency (P < 0.01). That sperm motility and acrosomal morphology were very acceptable immediately post-thaw and after 4h of incubation indicated the efficacy of cryopreserving ram spermatozoa under controlled conditions in medium-sized straws.

Sperm motion characteristics of Garole rams raised for a prolonged period in a semi-arid tropical environment.

The Garole is a prolific but less well known and rare breed of small sheep found in the hot and humid Sunderban region of West Bengal. An ability to breed throughout the year and to graze in knee-deep water, resistance to foot rot and a strong mothering instinct are some of the special features of this breed. Garole rams could provide germplasm to incorporate prolificacy traits by artificial insemination of the nonprolific sheep breeds found in abundance in the semi-arid and arid tropical climates of India. The aim of the present study was to evaluate the semen production by Garole rams maintained in a semi-arid climate for three years and to objectively assess their semen quality by a computer-assisted sperm analysis technique. The donor rams were randomly selected each year from the original flock procured from their natural habitat or from the offspring born at the Institute farm. Semen was collected weekly for three weeks each autumn for three consecutive years from 8 rams each year. The overall means (SD) of the traits that did not differ significantly with age or year were volume, concentration, curvilinear velocity, average path velocity, amplitude of lateral head displacement, beat frequency, motility and the percentages of rapid motile sperms and of slow motile sperms. The age of the rams had a significant effect (p<0.05) on the straight-line velocity but this was not significantly affected by the length of exposure to the semi-arid climate. However, the age and year had significant effects (p<0.05) on linearity, straightness and the percentage of medium motile sperms. It was concluded that Garole rams are capable of producing good-quality semen even after a prolonged period of exposure to a semi-arid tropical climate.

Effect of freezing temperature, at which straws were plunged into liquid nitrogen, on the post-thaw motility and acrosomal status of ram spermatozoa.

The present study was conducted to observe the effect of initial freezing temperature on subsequent survival and acrosomal integrity of Malpura and Bharat Merino ram spermatozoa during post-thawing incubation. Semen samples were diluted in TEST-yolk-glycerol extender, loaded in 0.25 ml straws and cooled down to -25, -75 or -125 degrees C freezing temperature using a programmable cell freezer. Computer assisted sperm analysis and acrosomal integrity of thawed samples were assessed after thawing and at hourly intervals during incubation at 37 degrees C for 4 h. The percentage of motile cells in samples frozen at -125 degrees C were 80.3 and 63.7 after post-thawing and -thawing incubation, compared to 75.9 and 39.7 at -25 degrees C or 73.9 and 51.8 at -75 degrees C temperatures, respectively. The spermatozoa with normal acrosome were also significantly, respectively, higher in samples frozen at -125 degrees C, compared to -25 and -75 degrees C temperatures. There were no significant breed variations on percentage of motile, percentage of rapidly motile cells, percentage of normal acrosomes, curvilinear velocity and lateral head displacement except straight line velocity and average path velocity of spermatozoa. The results indicated that -125 degrees C initial freezing temperature conferred the best cryopreserving ability to ram spermatozoa for post-thawing thermoresistance test compared to -25 or -75 degrees C freezing temperature.

Development and application of ovine reproductive technologies: an Indian experience.

Sheep play an important role in the Indian economy by providing employment to a large population of marginal and landless farmers. The production from native breeds is relatively low due to their poor reproductive efficiency. Embryo transfer technology can be utilised for faster multiplication of elite animal to increase the genetic gain. A great deal of research is involved to overcome the constraints in the technology, i.e. expensive and complicated nature of the technology and low success rate. In order to avoid surgical involvement, procedures of laparoscope aided embryo collection and transfer have been developed. Although the use of FSH of ovine origin has given more consistent superovulatory response, but it is still too low to get sufficient numbers of progeny from a donor ewe. The progress made in cryopreservation of ram semen has opened the possibility for conservation and utilisation of frozen semen of elite rams in sheep improvement programme. The lambing rate obtained after laparoscope aided intrauterine artificial insemination with frozen semen is encouraging but the impetus is now to develop the non-invasive transcervical insemination technique.

Oxidative degradation of triazine derivatives in aqueous medium: a radiation and photochemical study.

Pulse and steady state radiolysis techniques have been used to determine the bimolecular rate constants and to investigate the spectral nature of the intermediates and the degradation induced by hydroxyl radicals ((*)OH) with 1,3,5-triazine (T), 2,4, 6-trimethoxy-1,3,5-triazine (TMT), and 2,4-dioxohexahydro-1,3, 5-triazine (DHT) in aqueous medium. A competitive kinetic method with KSCN as the (*)OH scavenger was used to determine the rate constants for the reaction of (*)OH with T, TMT, and DHT. The bimolecular rate constants are 3.4 x 10(9), 2.06 x 10(8), and 1.61 x 10(9) dm(3) mol(-)(1) s(-)(1) respectively, for T, TMT, and DHT at pH approximately 6. The transient absorption spectra obtained from the reaction of (*)OH with T, TMT, and DHT have single absorption maxima at 320, 300, and 300 nm, respectively, and were found to undergo a second-order decay. The formation of TOH(*) [C(6)OH-N(5)-yl radical], TMTOH(*) [N(5)OH-C(6)-yl radical], and DHT(*) [C(6)-yl radical] is proposed from the initial attack of (*)OH with T, TMT, and DHT, respectively. A complete degradation of TMT (10(-3) mol dm(-3)) was obtained after absorbed doses of 5 kGy in N(2)O-saturated solutions and 16 kGy in aerated solutions. A similar degradation pattern was obtained with DHT in N(2)O-saturated solutions. Complete degradation was observed with an absorbed dose of 7 kGy. On the basis of the results from both pulse and steady state radiolysis, a possible reaction mechanism involving (*)OH-mediated oxidative degradation is proposed. A complete photodecomposition of DHT was also observed in the presence of ferric perchlorate using ultraviolet light at low pH. Photoinduced electron transfer between Fe(III) and DHT in the Fe(III)-DHT complex and subsequent formation of DHT(*) are proposed to be the major processes that lead to the complete degradation of DHT at pH 3.

Mechanism of biochemical action of substituted 4-methylbenzopyran-2-ones. Part 5: Pulse radiolysis studies on the antioxidant action of 7,8-diacetoxy-4-methylcoumarin.

7,8-Dihydroxy-4-methylcoumarin (1, DHMC) and 7,8-diacetoxy-4-methylcoumarin (2, DAMC) were shown to possess radical scavenging property and strongly inhibit membrane lipid peroxidation. Although free polyphenolic compounds are known to be antioxidants, the antioxidant action of the acetoxy compound DAMC was intriguing. Hence, pulse radiolysis studies were undertaken to explain the antioxidant action of DAMC. Accordingly, DAMC and DHMC were separately reacted with the system generating azide radicals and the resulting transient spectra were recorded. The spectra so obtained in both the cases demonstrated peak at 410 nm, characteristic of phenoxyl radical. The rate constants for the formation of phenoxyl radical from DHMC and DAMC were 34 x 10(8) M(-1) s(-1) and 6.2 x 10(8) M(-1) s(-1), respectively. We propose that the free radical mediated oxidation of DAMC initially produces a radical cation that loses an acetyl carbocation to yield the phenoxyl radical. It is possible to conclude that the mechanism of the antioxidant action of DAMC follows the pathway similar to that of DHMC involving the formation of a stable phenoxyl radical.

Oxidative damage induced by the fullerene C60 on photosensitization in rat liver microsomes.

We have examined the ability of a commonly used fullerene, C60, to induce oxidative damage on photosensitization using rat liver microsomes as model membranes. When C60 was incorporated into rat liver microsomes in the form of its cyclodextrin complex and exposed to UV or visible light, it induced significant oxidative damage in terms of (1) lipid peroxidation as assayed by thiobarbituric acid reactive substances (TBARS), lipid hydroperoxides and conjugated dienes, and (2) damage to proteins as assessed by protein carbonyls and loss of the membrane-bound enzymes. The oxidative damage induced was both time- and concentration-dependent. C60 plus light-induced lipid peroxidation was significantly inhibited by the quenchers of singlet oxygen ((1)O2), beta-carotene and sodium azide, and deuteration of the buffer-enhanced peroxidation. These observations indicate that C60 is an efficient inducer of peroxidation and is predominantly due to (1)O2. Biological antioxidants such as glutathione, ascorbic acid and alpha-tocopherol significantly differ in their ability to inhibit peroxidation induced by C60. Our studies, hence, indicate that C60, on photosensitization, can induce significant lipid peroxidation and other forms of oxidative damage in biological membranes and that this phenomenon can be greatly modulated by endogenous antioxidants and scavengers of reactive oxygen species.

Mechanisms of protection by buthionine sulphoximine against gamma-ray-induced micronuclei in polychromatic erythrocytes of mouse bone marrow.

The effect of pretreatment with buthionine sulphoximine (BSO) on the radiosensitivity of mouse bone marrow cells was studied using the in vivo micronucleus test. Varying concentrations of BSO were injected into mice by intraperitoneal injection 2 h before irradiation, and the frequency of micronuclei in polychromatic erythrocytes (MnPCEs) of bone marrow were scored. Treatment with BSO resulted in a significant reduction (41% at 20 mg/kg body weight) in the frequency of micronuclei induced by 1 Gy gamma-rays. Reduction was observed in cells sampled at 24, 30 and 48 h postirradiation with no apparent effect on the ratio of poly- to normo-chromatic erythrocytes in BSO-treated versus control groups. Glutathione levels in the bone marrow of BSO-treated animals 2 h after a single injection were found to be unaltered. The protective effect of BSO was not observed if it was given either immediately or 2 h after irradiation. Based on these and earlier findings it seemed as if BSO molecules may be involved in physicochemical reactions with reactive species generated in the system by irradiation. BSO showed relatively high reaction rate constants with hydroxyl radical (.OH, 2.5 x 10(9) dm3 mol-1s1, calculated on the basis of competition kinetics) and with singlet oxygen (1O2, 4.3 x 10(7) dm3 mol-1s-1 but a lower rate constant with hydrated electrons (< or = 5.0 x 10(6) dm3 mol-1s1). Based on half-life estimates, transients formed and potential for damage to biomolecules, .OH and 1O2 seemed to be the possible species responsible. In vitro studies reveal that BSO has significant abilities to protect DNA against single-strand breaks and lipid peroxidation induced by 1O2 in microsomal membranes. This supports our hypothesis that BSO may be involved in scavenging the reactive species generated and that besides .OH, 1O2 may also be a major player in radiation damage.

Addition of e-aq and H atoms to hypoxanthine and inosine and the reactions of alpha-hydroxyalkyl radicals with purines. A pulse radiolysis and product analysis study.

The reactions of hydrated electrons e-aq with hypoxanthine and inosine were followed using pulse radiolysis methods. In a neutral solution the electron adduct of inosine is immediately protonated at the heteroatoms of the purine ring by water (k >> 2.5 x 10(6)s-1) to give In(N,O-H).. These N,O-protonated intermediates have a single absorption maximum at 300 nm. In basic solution the protonation of the electron adduct of inosine by water leads to other intermediate products with an absorption maximum at 350 nm. These intermediates are believed to be the C-protonated electron adducts of inosine (In(N,O-H).). In (N,O-H). and In(C-H). differ strongly in their ability to reduce p-nitroacetophenone (PNAP). In(N,O-H). are strong reductants and reduce PNAP quantitatively to PNAP.-. Based on the pH dependence of PNAP.- yields, two types of tautomers of In(C-H). could be distinguished. One of the tautomers can reduce PNAP, albeit with slower rate than In(N,O-H)., the other tautomer has no reducing properties. The latter is the one with the higher pKa and therefore thermodynamically more stable. The absorption spectrum of the intermediates produced in the reaction of e-aq with hypoxanthine at neutral pH is very similar to that of In(N,O-H). with a maximum at 300 nm. However, no build-up at 350 nm was observed in basic solution as in the case of the electron adduct of inosine. The reaction of H atoms with inosine produces in basic solution intermediate radicals with the same absorption spectrum as the C-protonated electron adducts of inosine. It is suggested that both the reactions of e-aq and H. with inosine in basic solution produce the same radical, namely the H-adduct of inosine (In(C-H)) with the highest pKa. alpha-Hydroxyalkyl radicals were found to react very slowly with purine bases and nucleosides in neutral to basic solutions. In acidic solution their reactivity increases and a number of rate constants were determined by pulse radiolysis measurements at pH 0.4. The intermediates from the reaction of 2-hydroxy-2-propyl radicals with inosine could be observed pulse spectrometrically in neutral and in basic solutions. In basic solution this reaction leads to intermediates with the same absorption maximum at 350 nm as that of the H-adduct of inosine. Furthermore, the yield of acetone was found to increase strongly in basic pH.(ABSTRACT TRUNCATED AT 400 WORDS)

Solution photochemistry of thymine and uracil.

With dienes as specific triplet quenchers, it has been shown that the photodimerization of thymine in acetonitrile proceeds entirely through the triplet state and the photodimerization of uracil in acetonitrile or in water proceeds in part through the triplet state. The photohydration of uracil probably does not involve the triplet state. Efficiencies of intersystem crossing of thymine and uracil in acetonitrile were determined.