Vitellogenin genes in fish: differential expression on exposure to estradiol

Three types of vitellogenins (Vgs) namely vitellogenin A (VgA), vitellogenin B (VgB) and vitellogenin C (VgC) have been identified in fishes. The existence of VgA and VgB is reported in the Indian freshwater murrel Channa punctatus. Gene-specific primers were designed using available nucleotide sequences in National Centre for Biotechnology Information (NCBI), for amplification of VgA and VgB cDNA. Differential processing of Vgs is evident in many fishes. Adult male murrel expressed both the VgA and VgB genes when estradiol-17β (E₂) is injected in vivo and Vg levels in blood quantified by Enzyme linked immunosorbent assay (ELISA) showed a dose-related response in such treatments. Cultured hepatocytes on treatment with E₂, however, expressed only VgB as detected by RT-PCR, suggesting different regulatory mechanism for the VgA and VgB genes.     

Laser control of chemical reactions

Experiments show how product pathways can be controlled by irradiation with one or more laser beams during individual bimolecular collisions or during unimolecular decompositions. For bimolecular collisions, control has been achieved by selective excitation of reagent vibrational modes, by control of reagent approach geometry, and by control of orbital alignment. For unimolecular reactions, control has been achieved by quantum interference between different reaction pathways connecting the same initial and final states and by adjusting the temporal shape and spectral content of ultrashort, chirped pulses of radiation. These collision-control experiments deeply enrich the understanding of how chemical reactions occur.     

Composition and mineralogy of iron-manganese concretions from some soils of the Indo-Gangetic plain in Northwest India

Iron-manganese concretions from three reference soil series of the Indo-Gangetic plain of Punjab have been investigated for their composition, micromorphology and mineralogy. The data show that amounts of Fe, Mn, Co, Ni, Cu and Zn in the concretions are higher than in the surrounding soil matrix. The contents of Mn, Co and Ni increase and that of Fe decreases with the increasing size of concretions. Cu or Zn contents do not show any systematic differences with concretion size. A major portion of these elements is oxalate- and dithionite-extractable. A minor fraction is present in the silicate structure as well as in the pyrophosphate-extractable forms. The Fe and Mn compounds present in the concretions are x-ray amorphous. The concretions appear to have developed in situ.     

Differential heat sensitivity of two cool-season legumes,chickpea and lentil,at the reproductive stage,is associated with responses in pollen function,photosynthetic ability and oxidative damage

Increasing temperatures are adversely affecting various food crops, including legumes, and this issue requires attention. The growth of two cool-season food legumes, chickpea and lentil, is inhibited by high temperatures but their relative sensitivity to heat stress and the underlying reasons have not been investigated. Moreover, the high-temperature thresholds for these two legumes have not been well-characterised. In the present study, three chickpea (ICCVO7110, ICC5912 and ICCV92944) and two lentil (LL699 and LL931) genotypes, having nearly similar phenology with respect to flowering, were grown at 30/20°C (day/night; control) until the onset of flowering and subsequently exposed to varying high temperatures (35/25, 38/28, 40/30 and 42/32°C; day/night) in a controlled environment (growth chamber; 12 hr/12 hr; light intensity 750 µmol m⁻² s⁻¹; RH-70%) at 108 days after sowing for both the species. Phenology (podding, maturity) was accelerated in both the species; the days to podding declined more in lentil at 35/25 (2.8 days) and 38/28°C (11.3 days) than in chickpea (1.7 and 7.1 days, respectively). Heat stress decreased flowering–podding and podding–maturity intervals considerably in both the species. At higher temperatures, no podding was observed in lentil, while chickpea showed reduction of 14.9 and 16.1 days at 40/30 and 42/32°C, respectively. Maturity was accelerated on 15.3 and 12.5 days at 38/28°C, 33.6 and 34 days at 40/30°C and 45.6 and 47 days at 42/32°C, in chickpea and lentil, respectively. Consequently, biomass decreased considerably at 38/28°C in both the species to limit the yield-related traits. Lentil was significantly more sensitive to heat stress, with the damage—assessed as reduction in biomass, reproductive function-related traits (pollen viability, germination, pollen tube growth and stigma receptivity), leaf traits such as membrane injury, leaf water status, photochemical efficiency, chlorophyll concentration, carbon fixation and assimilation, and oxidative stress, appearing even at 35/25°C, compared with 38/28°C, in chickpea. The expression of enzymatic antioxidants such as superoxide dismutase, catalase, ascorbate peroxidase, glutathione reductase and non-enzymatic antioxidants declined remarkably with heat stress, more so in lentil than in chickpea. Carbon fixation (assessed as Rubisco activity) and assimilation (assessed as sucrose concentration, sucrose synthase activity) were also reduced more in lentil than in chickpea, at all the stressful temperatures, resulting in more inhibition of plant biomass (shoot + roots), damage to reproductive function and severe reduction in pods and seeds. At 38/28°C, lentil showed 43% reduction in biomass, while it declined by 17.2% in chickpea at the same time, over the control temperature (30/20°C). At this temperature, lentil showed 53% and 46% reduction in pods and seed yield, compared to 13.4% and 22% decrease in chickpea at the same temperature. At 40/30°C, lentil did not produce any pods, while chickpea was able to produce few pods at this temperature. This study identified that lentil is considerably more sensitive to heat stress than chickpea, as a result of more damage to leaves (photosynthetic ability; oxidative injury) and reproductive components (pollen function, etc.) at 35/25°C and above, at controlled conditions.     

Genotyping safflower (Carthamus tinctorius) cultivars by DNA fingerprints

Summary Carthamus tinctorius (2n = 2x = 24) (family Asteraceae), commonly known as safflower, is widely cultivated in agricultural production systems of Asia, Europe, Australia and the Americas as a source of high-quality vegetable and industrial oil. India ranks first in the production of safflower oil. Fourteen cultivars, widely cultivated in various agro-climatic regions of India, have been fingerprinted by RAPD, ISSR, and AFLP markers utilizing 36, 21 primers, and 4 primer combinations, respectively. On an individual assay basis, AFLP has proven to be the best marker system as compared with the other two markers applied as assessed by high discriminating power (0.98), assay efficiency index (33.2), marker index (18.2), resolving power (40.62), and genotype index (0.856). Thirty-six RAPD and 21 SSR primers could differentiate a maximum of eight and four cultivars, respectively, whereas, two AFLP primer combinations could fingerprint all the 14 cultivars. To understand genetic relationships among these cultivars, Jaccard's similarity coefficient and UPGMA clustering algorithm were applied to the three marker data sets. Mean genetic similarities ranged from 0.689 (AFLP) to 0.952 (ISSR). Correlation coefficient comparisons between similarity matrices and co-phenetic matrices obtained with the three markers revealed that AFLP displayed no congruence vis-a-vis RAPD and ISSR data. However, strong correlation was observed between RAPD and ISSR marker systems. This paper reports the start of molecular biology programme targeting nuclear genome of safflower, a major world oilseed crop about whose genetics very little is known.     

Diversity and relationships of multipurpose seabuckthorn (Hippophae L.) germplasm from the Indian Himalayas as assessed by AFLP and SAMPL markers

Seabuckthorn, a non-leguminous nodule bearing dioecious shrub, is a storehouse of neutraceutical, pharmaceutical and cosmetic usage. The 348 genotypes of Hippophae rhamnoides ssp. turkestanica, H. salicifolia and H. tibetana were collected from 194 locations at 46 major sites across?~1,500?km from north-east to north-west Himalayas, harboring one of the most harsh, highly variable climatic and ecological conditions and rugged rocky terrain in the range of 3,000–5,000?m altitude. Amplified fragment length polymorphism (AFLP) and selective amplification of microsatellite polymorphic loci (SAMPL) DNA markers were utilized to assess, the genetic diversity of total spectrum of Himalayan germplasm resources and interrelationships among Hippophae taxa. A total of 151, 50, and 41 AFLP loci were detected in Hippophae rhamnoides ssp. turkestanica, H. salicifolia and H. tibetana, respectively; of these, 92.6, 30.6 and 25.1% were polymorphic. For SAMPL, the number of markers and polymorphism for these species were, respectively, 77 (77.7%), 41 (41.4%), and 23 (23.2%). Nei’s genetic diversity and Shannon’s information index values revealed that populations of H. rhamnoides ssp. turkestanica in Sumur and Raling in Ladakh and Lahaul-Spiti, respectively, were the most diverse. In H. salicifolia and H. tibetana, the populations in Changu, and Takcha and Guling in Spiti valley, were the most diverse. In H. rhamnoides ssp. turkestanica, high level of interpopulation genetic diversity with the little intra-population diversity was accompanied by very low gene flow (Nm) range, estimated by AFLP (0.571–0.943) and SAMPL (0.321–0.726) markers. In H. rhamnoides ssp. turkestanica, the genotypes collected from Uttaranchal were found to be the least diverse. The exclusive characteristics of the nuclear genome in the Uttaranchal genotypes warrant new species rank more closer to H. salicifolia rather than to H. rhamnoides ssp. turkestanica. The present results also provide explicit evidence to suggest that both H. salicifolia and H. tibetana deserve species rank.     

SCINTIGRAPHY OF THE SACROILIAC JOINT REGION IN ASYMPTOMATIC RIDING HORSES: SCINTIGRAPHIC APPEARANCE AND EVALUATION OF METHOD

The sacroiliac region in 33 clinically normal riding horses was examined with scintigraphy to determine the normal scintigraphic appearance, to evaluate a semiquantitative method used to calculate a sacroiliac joint (SI joint) ratio, and to identify factors that affect the scintigraphic appearance of the pelvis. The scintigraphic examination included dorsal views of each SI joint region and one lateral view of the urinary bladder. Two predefined areas (the SI joint and the area between the tuber sacrale and the SI joint) were evaluated subjectively by comparing the intensity in each area to the intensity in the tuber sacrale, and a semiquantitative method was applied on the images to calculate an SI joint ratio. The thickness of the muscle mass overlying each SI joint was measured by using ultrasound. A corrected ratio was calculated based on a corrected pixel count for each SI joint. Of 29 evaluated horses, 28 had normal radiotracer uptake in the SI joints and 10 horses had symmetric normal radiotracer uptake in the area between the tuber sacrale and the SI joint. The mean SI joint ratio was 0.53 on both the left and right sides, and the mean corrected SI joint ratio was 2.15 on the left side and 2.13 on the right side. Factors that affect the scintigraphic appearance of the pelvis were identified and included attenuation, radioactive urine, and muscle symmetry. The muscle thickness ranged from 8-11 cm, causing 71-82% attenuation. Radioactive urine in the urinary bladder was located ventral to the SI joint region in 16 horses, and four of these were excluded from further evaluation because of risk of misinterpretation. A visual assessment of symmetry of the horses' gluteal muscle mass was compared with the ultrasound measurements. Fourteen horses (14/33) were regarded symmetric by both techniques. Soft tissue attenuation was found to severely compromise the result and indicated that only lesions in the SI joint with severely increased radiotracer uptake can be detected with scintigraphy. Knowledge about presence of radioactive urine ventral to the SI joint region and assessment of muscle symmetry is essential for a correct subjective evaluation. Any situation with difference in muscle mass between the left and right side of the pelvis will give a false impression of increased radiotracer uptake on the side with lesser muscle mass.     

Further heterogeneity of human alpha interferon mRNA species

Translationally active (in Xenopus oocytes) human alpha interferon (IFN) messenger RNA's (mRNA's) derived from Sendai virus--induced leukocyte cultures display a bimodal distribution of RNA lengths on electrophoresis through agarose-CH3HgOH gels. The major population (alpha s) consists of mRNA of length 0.7 to 1.4 kilobases, while the minor population (alpha L) consists of RNA of length 1.6 to 3.5 kilobases. Induction of human leukocytes in the presence of 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB; 100 micromolar) appears to inhibit the accumulation of IFN-alpha s and to enhance that of IFN-alpha L mRNA's (average length about 1.8 kilobases in preparations from DRB-treated cells). Interferons derived from the alpha s mRNA's represent the group of previously recognized alpha interferons while the alpha L interferons are distinguishable from this group by their lower heterospecific activity on bovine cells compared to human cells, their apparent slower mobility in sodium dodecyl sulfate--polyacrylamide gels, and their apparent heteroclitic response toward an antiserum to IFN-alpha.     

Results of Transsphenoidal Hypophysectomy in 52 Dogs With Pituitary-Dependent Hyperadrenocorticism

Objective —To evaluate microsurgical transsphenoidal hypophysectomy in dogs with pituitary-dependent hyperadrenocorticism (PDH). Study Design —Prospective study to evaluate the results (survival and disease-free interval, remission, recurrence) and complications of microsurgical transsphenoidal hypophysectomy by clinical follow-up, computed tomography (CT), and urinary corticoid-to-creatinine (C/C) ratios in dogs with PDH. The effect of surgical experience was investigated by comparing results of hypophysectomy cases 1 through 26 and 27 through 52. Animals or Sample Population—52 dogs with PDH. Results —Preoperative CT enabled accurate assessment of pituitary size (24 nonenlarged and 28 enlarged) and localization relative to intraoperative anatomic landmarks. Treatment failures included procedure-related mortalities (five dogs) and incomplete hypophysectomies (four dogs). The 1-year estimated survival rate was 84% (95% confidence interval [CI], 71% to 92%). The 2-year estimated survival rate was 80% (95% CI, 65% to 90%). In 43 dogs, the hyperadrenocorticism went into remission. Hyperadrenocorticism recurred in five dogs. The 1-year estimated relapse-free fraction was 92% (95% CI, 76% to 97%). The main complications were transient, mild, postoperative hypernatremia; transient reduction or cessation of tear production (25 eyes in 18 dogs); permanent (five dogs) or prolonged (nine dogs) diabetes insipidus; and secondary hypothyroidism. Normal tear production had resumed in all but one case after a median period of 10 weeks. In the second case series (27 through 52), the hospitalization period was shorter, the number of dry eyes fewer, the survival fraction greater, and the postoperative mortality lower than in the first series. In 15 dogs in which remission was obtained, postoperative CT images suggested the presence of small pituitary remnants; in 1 of these, hyperadrenocorticism recurred. In 46 dogs, the histological diagnosis was pituitary adenoma. Conclusions —Microsurgical transsphenoidal hypophysectomy in dogs with PDH is an effective method of treatment in specialized veterinary institutions having access to advanced pituitary imaging techniques. Postoperative CT findings do not correlate well with remission or subsequent recurrence of hyperadrenocorticism. Clinical Relevance —The neurosurgeon performing hypophysectomies must master a learning curve and must be familiar with the most frequent complications of the operation to recognize them as early as possible and to treat them immediately and effectively. Urinary C/C ratios are sensitive indicators for the assessment of remission and recurrence of hyperadrenocorticism.