How does a re-colonizing population of Asian elephants affect the forest habitat?

The Asian elephant Elephas maximus is currently re-colonizing the Bardia National Park in lowland Nepal. We studied their impact on woody vegetation in the nutrient-rich floodplain and in the relatively nutrient-poor sal forest. The types and extent of tree impact were recorded along fixed-width transects (335 km). Species composition, density and size classes ≥8 cm diameter breast height (dbh) were recorded in 15-m radius random plots ( n =95). Impact was higher in the floodplain complex than in the sal-dominated forest. Our hypothesis that elephants were more selective on species in the nutrient-poor sal forest was only partly supported; the niche breadth of impacted trees was slightly higher in the floodplain complex. Pushed-over trees accounted for the highest proportion of impact (55%), followed by killed trees (39%). Of the pushed trees, 10% were not used for food. Among food trees, elephants selectively impacted size class 12–16 cm dbh, whereas non-food trees were impacted independently of size. A large proportion of the freshly browsed trees had been felled previously, indicating that most felled trees survived, enabling elephants to feed on them again. This may reflect an evolutionary adaptation among long-lived species with high site fidelity. Owing to preferential use but low abundance, two species in sal forest, Grewia spp. and Desmodium oojeinense , were found to be particularly vulnerable to local extinction due to elephants. Although the elephants had impacted a large number of species (62, 73% of all), 56.4% of the impacted trees consisted of Mallotus phillippinensis . A recently observed increase in the density of M. phillippinensis and the concurrent reduction of the hardly utilized Shorea robusta indicates that the rapidly growing elephant population may modify the composition of the forest by increasing its preferred food species.     

Dephosphorization of LD slag by phosphorus solubilising bacteria

Phosphorus is one of the major nutrients, and microbial solubilisation of insoluble mineral phosphate in soil is an important process in natural ecosystem and in agricultural soil. Many soil microorganisms display the ability to solubilize many insoluble inorganic phosphates. They are generally referred as phosphorus solubilising microorganisms (PSM). In this study an attempt was made to look into the phosphorus solubilisation efficiency of some commonly available soil bacteria and their possible application in bio-beneficiation of metallurgical waste like LD Slag. Linz -Donawitz (LD) slag is produced in large quantities (200 kg LD slag per ton of hot metal) and poses a substantial disposal problem in the iron and steel making industry. LD slag contains around 29% Ca, 21% Fe, and 5% Mg. Its phosphorus content is about 1.5-6%. Due to presence of high amount of Ca, it can be used as flux in blast furnace, but presence of high amount of phosphorus in the LD slag makes them unsuitable for industrial application. Removal of phosphorus with the help of phosphorus solubilising microorganisms may be a great advantage in biotechnological applications. Two gram positive bacteria belonging to genus Bacillus and two gram negative bacteria belonging to genus Pseudomonas were selected in this study. Phosphorus solubilisation efficiency was studied initially with tricalcium phosphate as model insoluble phosphate compound at different sugar concentration, NaCl concentration and at different initial pH of the medium. About 35% of ‘P’ could be solubilized from LD slag by Pseudomonas aeruginosa at 2% solid content.     

Efficient plant regeneration from cell suspension-derived callus of East Indian rosewood (Dalbergia latifolia Roxb.)

A procedure is outlined for the establishment of a proliferating cell suspension culture of East Indian rosewood (Dalbergia latifolia Roxb.) and efficient plant regeneration from callus derived from such cultures. Callus was induced from hypocotyl segments derived from 1-week-old axenic seedlings on Murashige and Skoog (1962) medium (MS) containing 10.8 μM naphthaleneacetic acid (NAA) and 2.2 μM benzyladenine (BA). Calli were increased by subculturing on MS supplemented with same growth regulators and 10% coconut water (CW). Friable calli were used to initiate cell suspension cultures. Optimum cell proliferation occurred in MS containing 10.8 μM NAA, 2.2 μM BA and 10% CW, using an initial inoculum cell density of 2%. Cell clumps composed of 20–25 cells harvested from suspension cultures at the exponential growth phase readily formed callus within 3 weeks following plating on the semi-solid MS as above. High-frequency shoot-bud differentiation was induced in these calli on MS containing 2.7 μM NAA and 13.3 μM BA. The regeneration frequency declined at higher BA concentrations. The organogenic potential of the cell suspensions was influenced by the age of the culture. Regenerated shoots were rooted on half-strength MS containing 5.7 μM indole-3-acetic acid, 4.9 μM indole-3-butyric acid and 5.3 μM indole-3-propionic acid. The plantlets were acclimatized and established in soil. Received: 22 August 1997 / Revision received: 21 May 1998 / Accepted: 1 June 1998     

Biogenic synthesis of floral-shaped gold nanoparticles using a novel strain,Talaromyces flavus

A biogenic route was adopted towards the synthesis of gold nanoparticles using the extract of a novel strain, Talaromyces flavus. Reduction of chloroauric acid by the fungal extract resulted in the production of gold nanoparticle, which was further confirmed by the concordant results obtained from UV–visible spectroscopy, energy dispersive spectroscopy (EDS), and dynamic light scattering (DLS) analysis. Morphology and the crystal nature of the synthesized nanoparticles were characterized using transmission electron microscopy (TEM), X-ray diffraction (XRD) and selected area electron diffraction (SAED). A direct correlation was observed between nanoparticle formation and the concentration of reducing agent present in the fungal extract. The time-dependent kinetic study revealed that the bioreduction process follows an autocatalytic reaction. Crystalline, irregular, and mostly flower-shaped gold nanoparticles with a mean hydrodynamic radius of 38.54?±?10.34 nm were obtained. pH played a significant role on production of mono-dispersed nanoparticle. FTIR analysis partially deciphered the involvement of –NH₂, ?SH, and –CO groups as the probable molecules in the bio-reduction and stabilization process. Compared to the conventional methods, a time-resolved, green, and economically viable method for floral-shaped nanoparticle synthesis was developed.     

Substrate specificity and kinetic mechanism of mammalian G9a histone H3 methyltransferase

Lysine-specific murine histone H3 methyltransferase, G9a, was expressed and purified in a baculovirus expression system. The primary structure of the recombinant enzyme is identical to the native enzyme. Enzymatic activity was favorable at alkaline conditions (>pH 8) and low salt concentration and virtually unchanged between 25 and 42 degrees C. Purified G9a was used for substrate specificity and steady-state kinetic analysis with peptides representing un- or dimethylated lysine 9 histone H3 tails with native lysine 4 or with lysine 4 changed to alanine (K4AK9). In vitro methylation of the H3 tail peptide resulted in trimethylation of Lys-9 and the reaction is processive. The turnover number (k(cat)) for methylation was 88 and 32 h(-1) on the wild type and K4AK9 histone H3 tail, respectively. The Michaelis constants for wild type and K4AK9 ((K(m)(pep))) were 0.9 and 1.0 microM and for S-adenosyl-L-methionine (K(m)(AdoMet)) were 1.8 and 0.6 microM, respectively. Comparable kinetic constants were obtained for recombinant histone H3. The conversion of K4AK9 di- to trimethyl-lysine was 7-fold slower than methyl group addition to unmethylated peptide. Preincubation studies showed that G9a-AdoMet and G9a-peptide complexes are catalytically active. Initial velocity data with peptide and S-adenosyl-L-methionine (AdoMet) and product inhibition studies with S-adenosyl-L-homocysteine were performed to assess the kinetic mechanism of the reaction. Double reciprocal plots and preincubation studies revealed S-adenosyl-L-homocysteine as a competitive inhibitor to AdoMet and mixed inhibitor to peptide. Trimethylated peptides acted as a competitive inhibitor to substrate peptide and mixed inhibitor to AdoMet suggesting a random mechanism in a Bi Bi reaction for recombinant G9a where either substrate can bind first to the enzyme, and either product can release first.     

GI side-effects of a possible therapeutic GRF analogue in monkeys are likely due to VIP receptor agonist activity.

Growth hormone (GH) is used or is being evaluated for efficacy in treatment of short stature, aspects of aging, cardiac disorders, Crohn's disease, and short bowel syndrome. Therefore, we synthesized several stable growth hormone-releasing factor (GRF) analogues that could be therapeutically useful. One potent analog, [D-Ala(2),Aib(8, 18,)Ala(9, 15, 16, 22, 24-26,)Gab(27)]hGRF(1-27)NH(2) (GRF-6), with prolonged infusion caused severe diarrhea in monkeys; however, it had no side-effects in rats. Because GRF has similarity to VIP/PACAP and VIPomas cause diarrhea, this study investigated the ability of this and other GRF analogues to interact with the VIP/PACAP receptors. Rat VPAC(1)-R (rVPAC(1)-R), human VPAC(1)-R (hVPAC(1)-R), rVPAC(2)-R and hVPAC(2)-R stably transfected CHO and PANC 1 cells were made and T47D breast cancer cells containing native human VPAC(1)-R and AR4-2J cells containing PAC(1)-R were used. hGRF(1-29)NH(2) had low affinity for both rVPAC(1)-R and rVPAC(2)-R while VIP had a high affinity for both receptors. GRF-6 had a low affinity for both rVPAC(1)-R and rVPAC(2)-R and very low affinity for the rPAC(1)-R. VIP had a high affinity, whereas hGRF(1-29)NH(2) had a low affinity for both hVPAC(1)-R and hVPAC(2)-R. In contrast GRF-6, while having a low affinity for hVPAC(2)-R, had relatively higher affinity for the hVPAC(1)-R. In guinea pig pancreatic acini, all GRF analogues were full agonists at the VPAC(1)-R causing enzyme secretion. These results demonstrate that in contrast to native hGRF(1-29)NH(2,) GRF-6 has a relatively high affinity for the human VPAC(1)-R but not for the human VPAC(2)-R, rat VPAC(1)-R, rat VPAC(2)-R or rat PAC(1)-R. These results suggest that the substituted GRF analog, GRF-6, likely causes the diarrheal side-effects in monkeys by interacting with the VPAC(1)-R. Furthermore, they demonstrate significant species differences can exist for possible therapeutic peptide agonists of the VIP/PACAP/GRF receptor family and that it is essential that receptor affinity assessments be performed in human cells or from a closely related species.     

Correlation of behavioral and neurochemical effects of acute administration of cocaine in rats.

Effects of cocaine were investigated on spontaneous motor activity (SMA) and stereotypy as well as on the concentrations of norepinephrine (NE), dopamine (DA), serotonin (5-HT) and acetylcholine (ACh) in the discrete brain areas, such as the caudate nucleus (CN), diencephalon-midbrain (DM) and pons-medulla (PM) in rats up to 90–120 min following its injection in single doses (15–20 mg/kg, i.p.). After cocaine administration, the SMA was increased usually reaching its peak between 10–20 min, and then decreased gradually. Stereotypy and its components gradually increased to their maximum at about 50–60 min and remained at that level during rest of 120 min sessions. NE levels slightly increased in the DM and PM at 10 min post-drug after which they were decreased at 20 min. DA levels in the CN and DM were increased markedly at 20 min post-drug and decreased at 40 min. 5-HT levels in DM and PM decreased gradually up to 20 min, then began to increase. ACh level in the CN was gradually increased at 40 min and then decreased. It appears that cocaine-induced hyperactivity and stereotypy followed release of NE and DA after their accumulation in the respective brain areas.     

Three major forms of adenylate kinase from adult and fetal rat tissues.

The major enzymatic forms of adenylate kinase have been purified to homogeneity from fetal liver and adult brain of the rat. The two enzymes differ with respect to isoelectric points, Km (ATP), Km (AMP), and Ka (citrate). Antibody to adult liver adenylate kinase does not inhibit either enzyme, while entibody to adult skeletal muscle enzyme inhibits the brain enzyme but not the fetal liver enzyme. It is therefore probable that there are three major forms of adenylate kinases in fetal and adult rat tissues.     

Evidence for excision repair in promitochondrial DNA of anaerobic cells of Saccharomyces cerevisiae.

The respiratory adaptation (i.e., essentially mitochondrial biogenesis) in the excision repair-defective rad3-type mutants of Saccharomyces cerevisiae undergoing transition from the anaerobic to the aerobic state is found to be far more sensitive to 254-nm ultraviolet radiation (UV) than that of the RAD wild-type strain. We confirm that mitochondria of aerobic cells of a RAD strain lack the excision repair capacity of UV-induced pyrimidine dimers at all doses tested (1-15 J/m2). In contrast, in promitochondria of anaerobic cells of the wild-type strain excision repair appears to take place. This process is very efficient at low doses (at 0.5-5 J/m2 100% of the UV endonuclease-sensitive sites disappear), whereas at high doses its efficiency is reduced by about 50%. The promitochondrial excision repair of pyrimidine dimers appears to be under nuclear control since it is blocked in the rad2 mutant. Finally photoreactivation is found to be operating in nuclei, mitochondria and promitochondria.