Effect of home processing on total and extractable calcium and zinc content of spinach (Spinach oleracia) and amaranth (Amaranthus tricolor) leaves

Spinach (Spinacia oleracia) and amaranth (Amaranthus tricolor) leaves were stored in polyethylene bags and without packing for 24 and 48 hours in a refrigerator at 5 and 30 °C in polyethylene bags. The fresh leaves were also dried (oven and sun); blanched (5, 10 and 15 min) and cooked in an open pan and a pressure cooker. The processed leaves were analysed for total and extractable calcium and zinc content. The Ca and Zn content of these leaves varied from 1320 to 2120 and 11.70 to 12.60 mg/100 g DM and the percentage HCl-extractability was 77.82 to 81.92 and 85.16 to 86.15, respectively. No significant effects of drying and storage were observed on total Ca and Zn content and HCl-extractability while blanching and cooking resulted in significant improvement of HCl-extractibility of these two minerals. Thus, cooking and blanching are good ways to improve the HCl-extractibility of Ca and Zn.     

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.     

Alcohol self-administration disrupts reproductive function in female macaque monkeys

Female macaque monkeys self-administered high doses of alcohol (2.9 to 4.4 grams per kilogram per day) for 3 to 6 1/2 months. Amenorrhea, atrophy of the uterus, decreased ovarian mass, and significant depression of luteinizing hormone levels were associated with chronic alcohol intoxication. Reproductive system failure in female primates following self-induced dependence on alcohol parallels the results of clinical studies of alcoholic women.     

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.     

Monocyte-derived human B-cell growth factor identified as interferon-beta 2 (BSF-2, IL-6)

Soluble products of either Epstein-Barr virus (EBV)-infected B cells or activated monocytes promote the proliferation of EBV-infected B cells and permit their growth at low cell densities. This suggests that growth factors are important for B-cell immortalization by EBV. In this study, a monocyte-derived factor that promotes the growth of EBV-infected b cells was purified and identified as interferon-beta 2 (IFN-beta 2), which is also known as 26-kilodalton protein, B-cell differentiation factor (BSF-2), and interleukin-6 (IL-6). The purified protein has a specific activity of approximately 4 X 10(7) units per milligram of protein in assays of B-cell growth. Thus, IFN-beta 2/BSF-2 is a B-cell growth factor that promotes the proliferation of human B cells infected with EBV.     

Receptor-mediated drug delivery to macrophages in chemotherapy of leishmaniasis

Methotrexate coupled to maleylated bovine serum albumin was taken up efficiently through the "scavenger" receptors present on macrophages and led to selective killing of intracellular Leishmania mexicana amazonensis amastigotes in cultured hamster peritoneal macrophages. The drug conjugate was nearly 100 times as effective as free methotrexate in eliminating the intracellular parasites. Furthermore, in a model of experimental cutaneous leishmaniasis in hamsters, the drug conjugate brought about more than 90% reduction in the size of footpad lesions within 11 days. In contrast, the free drug at a similar concentration did not significantly affect lesion size. These studies demonstrate the potential of receptor-mediated drug delivery in the therapy of macrophage-associated diseases.     

Isolation of T-cell tropic HTLV-III-like retrovirus from macaques

The isolation of a T-cell tropic retrovirus from three immunodeficient macaques and one macaque with lymphoma is described. The morphology, growth characteristics, and antigenic properties of this virus indicate that it is related to the causative agent of acquired immune deficiency syndrome in humans (HTLV-III or LAV). This virus is referred to as simian T-lymphotropic virus type III (STLV-III) of macaques. The existence of a cytopathic, T-cell tropic virus resembling HTLV-III in monkeys may facilitate study of disease induction and vaccine development in an animal model.     

In vitro induction of vitellogenin by estradiol 17 β in isolated hepatocytes of catfish, Clarias gariepinus

Vitellogenin is a female-specific calcium-binding glycolipophosphoprotein synthesized in the hepatocytes of fishes. Its synthesis can be induced in fishes of either sex by estradiol or by xenoestrogens. To study the in vitro synthesis of vitellogenin, different culture conditions were set up using the hepatocytes of Clarias gariepinus. The present study reports on a non-enzymatic procedure for isolation and culture of hepatocytes from the liver of the catfish Clarias gariepinus, in order to study the effects of estradiol on vitellogenin synthesis in vitro. The procedure employs chelating properties of ethylenediamine tetracetic acid to achieve cell viability in excess of 95%. Equal numbers of isolated cells were incubated in different culture media viz. RPMI F1640, Medium-199, and Williams’ Medium E. At 36 h, cell attachment and monolayer formation is faster in M-199 and Williams’ Medium E than in RPMI. In order to study the effects of estradiol on vitellogenin synthesis, the isolated hepatocytes were seeded in Williams’ Medium E in 24-well cell culture plates. 17 β-estradiol (E₂) was introduced in the culture plates at different concentrations and for different time periods. The media were assayed for vitellogenin using competitive ELISA. Vitellogenin appeared in the medium after 48 h of incubation with 10⁻⁵ M estradiol whereas after 72 h of incubation 5×10⁻⁷ M E₂ could elicit the synthesis.     

First glimpse into the origin and spread of the Asian longhorned tick,Haemaphysalis longicornis,in the United States

Established populations of Asian longhorned ticks (ALT), Haemaphysalis longicornis, were first identified in the United States (US) in 2017 by sequencing the mitochondrial cytochrome c oxidase subunit I (cox1) ‘barcoding’ locus followed by morphological confirmation. Subsequent investigations detected ALT infestations in 12, mostly eastern, US states. To gain information on the origin and spread of US ALT, we (1) sequenced cox1 from ALT populations across 9 US states and (2) obtained cox1 sequences from potential source populations [China, Japan and Republic of Korea (ROK) as well as Australia, New Zealand and the Kingdom of Tonga (KOT)] both by sequencing and by downloading publicly available sequences in NCBI GenBank. Additionally, we conducted epidemiological investigations of properties near its initial detection locale in Hunterdon County, NJ, as well as a broader risk analysis for importation of ectoparasites into the area. In eastern Asian populations (China/Japan/ROK), we detected 35 cox1 haplotypes that neatly clustered into two clades with known bisexual versus parthenogenetic phenotypes. In Australia/New Zealand/KOT, we detected 10 cox1 haplotypes all falling within the parthenogenetic cluster. In the United States, we detected three differentially distributed cox1 haplotypes from the parthenogenetic cluster, supporting phenotypic evidence that US ALT are parthenogenetic. While none of the source populations examined had all three US cox1 haplotypes, a phylogeographic network analysis supports a northeast Asian source for the US populations. Within the United States, epidemiological investigations indicate ALT can be moved long distances by human transport of animals, such as horses and dogs, with smaller scale movements on wildlife. These results have relevant implications for efforts aimed at minimizing the spread of ALT in the United States and preventing additional exotic tick introductions.     

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.