Quantification of Parasite Load in Clinical Samples of Leishmaniasis Patients: IL-10 Level Correlates with Parasite Load in Visceral Leishmaniasis

A rapid and accurate method to detect and quantify Leishmania parasite is urgently needed to facilitate early diagnosis of Leishmaniasis and monitoring of antileishmania therapy. In this study, real-time assay was applied to estimate parasite load in clinical samples of visceral leishmaniasis (VL) and post kala-azar dermal leishmaniasis (PKDL) patients. The mean parasite load in blood of VL patients (n = 31) was 8,372 parasites/ml, while the mean parasite load in bone marrow aspirate (BMA) was 194,962 parasites/million nucleated cells (n = 12). Parasite load was undetectable after treatment with amphotericin B (n = 9) in VL, while a residual parasite burden was detected in 2 of 6 patients following treatment with sodium antimony gluconate. Further, circulating levels of IFN-γ, TNF-α, IL-10, IL-6, IL-4 and IL-2 were analysed in VL patients (n = 29) by Cytometric Bead Array to evaluate correlation with parasitic load. Interestingly, IL-10 levels correlated significantly with parasite load (r = 0.82, P<0.0001). The mean parasite load in dermal lesions of PKDL patients was 9,502 parasites/µg tissue DNA at pre-treatment stage (n = 25), with no detectable parasites after therapy (n = 5). Parasite burden was distinctly higher (P<0.0001) in nodular lesions (n = 12) (19,586 parasites/µg tissue DNA) compared to papular/macular lesions (n = 13, 193 parasites/µg tissue DNA). Further, chronic PKDL lesions showed significantly (P = 0.0166) higher parasite load in comparison with acute lesions. Results indicate that chronic, nodular cases constitute the major parasite reservoir for anthroponotic transmission. Our results establish that the high parasite load in VL is strongly correlated with a high level of IL-10, implicating IL-10 as a marker of disease severity. The assay is applicable for diagnosis as well as prognosis of both VL and PKDL, providing a simple molecular tool to monitor the efficacy of antileishmanial drugs or vaccines.     

Establishment and growth of Quercus floribunda seedlings after a mast year

Abstract. Population dynamics, mortality factors, growth and dry weight changes in Quercus floribunda seedlings were studied to determine the implication of sun and shade microsites and the role of mast years on regeneration and maintenance of oak forests in Kumaun Himalaya. The number of seedlings was significantly greater at the sun microsites, but seedling survival was not, as ground herbage clearance at sun microsites accounted for 53.5% mortality. Insect herbivores (e.g. acorn worm) were also one of the major mortality factors. At both microsites, most mortality was encountered between June and August. The initial seedling height at sun microsites was significantly lower compared to shade microsites. Seedlings at sun microsites were superior in terms of number of leaves and peak dry weight. Dry mass allocation in seedling components and root:shoot ratio was similar at both microsites. The study shows the importance of sun microsites and of masting in the potential for regeneration and expansion of Q. floribunda forests.     

Daily Increments of Light Hours Near Vernal Equinox Synchronize Circannual Testicular Cycle of Tropical Spotted Munia

In some long-lived organisms, particularly in tropical birds and migrants that spend part of the year close to the equator, endogenous circannual rhythms have been demonstrated in seasonal events like reproduction, molt, and migration. These, like the circadian rhythms, are expressed only in constant conditions of illumination with a periodicity deviating from 1 yr. If birds followed this periodicity, they would soon be out of phase with the annual calendar and perish and, therefore, they would need to be synchronized. However, almost nothing is known as to how synchronization is achieved in birds. Herein, with the help of a suitable model, viz., the tropical spotted munia and long-term experiments conducted in series over a 5-yr period, we provide direct evidence for the first time indicating that the segment of annual photocycle with maximal rate of increase prior to vernal equinox (approximately between mid-February and mid-March) synchronizes the circannual reproductive cycle with the monsoon period of ample food supply through a phase delay. Data also indicate, contrary to the prevalent view, that birds in the tropics can perceive minor changes in day-length, that birds respond to progressive changes in day-length as distinct from responding to fixed photoperiods of particular durations, and that birds can actually distinguish the quality of the environmental signal, viz., vernal equinox from early spring, or increasing days of spring from decreasing days of autumn. The underlying mechanisms, although yet to be identified, appear to involve a gonado-inhibitory rather than the conventional gonado-stimulatory response to increasing day-length. The photoperiodic responses of spotted munia are distinctly different from that of any “long-day” birds described thus far and do not conform to the prevalent “circadian coincidence” hypothesis of photoperiodism.     

Amide derivatives of 9,11-seco-estra-1,3,5(10)-trien-11-oic acid as modified orally active estrogen agonists with moderate antagonistic activity

Synthesis of amide derivatives of 9,11-seco-estra-1,3,5(10)-trien-11-oic acid containing alkyl and aromatic amine residues has been carried out with an aim to prepare orally active estrogen antagonists. Modification of the estradiol molecule in the form of C-seco-amide derivatives has led to their high oral absorption. Compounds 7 an n-propyl amide, 8 an n-butyl amide, and 16 a p-anisidyl amide of C-seco-estrane showed significant estrogen antagonistic activity (>20%), while, the majority of compounds possessed high estrogen agonistic activity on oral administration at 10mg/kg dose in rats.     

Synthesis of a novel plant growth promoter from gallic acid

Gallic acid has been modified to naphthophenone derivatives with esterified fatty acid side chain. Compound 12, an ethyl crotonate ester of naphthophenone derivative has shown potent auxin like growth promoter activity. This is the first example of naphthophenone derivatives with plant growth promoting activity.     

Screening interspecific hybrids of<Emphasis Type="Italic"> Populus</Emphasis> (<Emphasis Type="Italic">P. ciliata</Emphasis> ×<Emphasis Type="Italic"> maximowiczii</Emphasis>) using AFLP markers

Hybrids of Populus ciliata × maximowiczii are very vigorous and outperform both the parents in growth performance and yield. Genetic evaluation of 24 of these interspecific hybrids along with the two mother trees (Populus ciliata), and five male-parent (Populus maximowiczii) genotypes was carried out using the AFLP marker assay. Eight AFLP primer combinations detected 428 markers, of which 280 (66%) were polymorphic. Genetic relationships within the samples were evaluated by generating the similarity matrix based on Jaccards coefficient. The phenetic dendrograms, as well as the PCO plots, separated the hybrids and the two parent species into three distinct clusters. The hybrids grouped closer to the P. ciliata (female parent) cluster as compared to the P. maximowiczii (male parent) cluster. The hybrid cluster contained internal groupings, which correlated to some extent with growth performance. The four best performing hybrids (42m1, 65m1, 23m2, Cm2-5-20/91) formed a distinct sub-cluster. Data from a single primer combination was sufficient for distinguishing the hybrids from the parents and assigning paternity. The hybrids showed 22 markers that were absent in P. ciliata but were monomorphically present in all the hybrids, suggesting outcrossing and common paternity. Further, these 22 markers were found in all the P. maximowiczii genotypes confirming it as the male parent. These male-specific markers can be converted to SCAR markers and used for rapid screening of the P.ciliata × maximowiczii hybrids. The primer combination E-AAC × M-CAA was identified as most suitable for ascertaining true hybridity. AFLP proves to be a useful tool for screening of P. ciliata × maximowiczii hybrids at the early stages of development.Communicated by H.F. Liskens