Response of particle-associated bacteria to long-term heavy metal contamination in a tropical estuary.

Estuaries being the connecting link between terrestrial and marine environment, experience spatial variations in the hydrographic variables as well as concentrations of pollutants. The present study reports a contrasting difference in the metal tolerance and enzyme activity of particle-associated bacteria (PAB) isolated from the upstream and downstream reaches of a tropical estuary [Cochin Estuary (CE) in the southwest coast of India], exposed to different levels of heavy metal contamination. The upstream of the estuary has been overloaded with heavy metals in the last few decades, while the downstream is less polluted. There were only 25% of culturable PAB phylogenetically common in both upstream and downstream. The PAB isolated from the upstream were dominated by γ-proteobacteria (48.1%) followed by α-proteobacteria (25.0%), while it was in the reverse order of α-proteobacteria (45.9%) and γ-proteobacteria (36.1%) in the downstream. More number of PAB from the upstream showed tolerance to higher concentrations of Zn and Cd. The Acinetobacter sp. MMRF1051 isolated from the upstream showed tolerance up to 250 mM Zn, 100 mM Cd, and 250 mM Ni. The enzyme expression profile of PAB from downstream was in the order of lipase > phosphatase > ß-glucosidase > aminopeptidase, while it was in the order of ß-glucosidase > lipase > aminopeptidase > phosphatase in the upstream of the estuary. The present study shows the selective pressure exerted by heavy metal pollution on the diversity of culturable bacteria associated with particulate matter in a tropical estuary. Also, the variation in their enzyme activities may impinge the remineralization of particulate organic matter (POM) in the system and may impart adverse impacts on ecosystem functioning.

Role of heavy metals in structuring the microbial community associated with particulate matter in a tropical estuary.

Particulate matter (PM), which are chemically and biochemically complicated particles, accommodate a plethora of microorganisms. In the present study, we report the influence of heavy metal pollution on the abundance and community structure of archaea and bacteria associated with PM samples collected from polluted and non-polluted regions of Cochin Estuary (CE), Southwest coast of India. We observed an accumulation of heavy metals in PM collected from CE, and their concentrations were in the order Fe > Zn > Mn > Cr > Pb > Cu > Cd > Co > Ni. Zinc was a major pollutant in the water (4.36-130.50 µgL-1) and in the particulate matter (765.5-8451.28 µgg-1). Heavy metals, Cd, Co, and Pb were recorded in the particulate matter, although they were below detectable limits in the water column. Statistical analysis showed a positive influence of particulate organic carbon, nitrogen, PM-Pb, PM-Zn and PM-Fe on the abundance of PM-archaea and PM-bacteria. The abundance of archaea and bacteria were ten times less in PM compared with planktonic ones. The abundance of PM-archaea ranged between 4.27 and 9.50 × 107and 2.73 to 3.85 × 107 cellsL-1 respectively for the wet and dry season, while that of PM-bacteria was between 1.14 and 6.72 × 108 cellsL-1 for both seasons. Community structure of PM-bacteria varied between polluted and non-polluted stations, while their abundance does not show a drastic difference. This could be attributed to the selective enrichment of bacteria by heavy metals in PM. Such enrichment may only promote the growth of metal resistant archaea and bacteria, which may not participate in the processing of PM. In such cases, the PM may remain without remineralization in the system arresting the food web dynamics and biogeochemical cycles.

Amlamax in the management of dyslipidemia in humans.

Hypercholesterolemia is the major cause of cardiovascular diseases leading to myocardial infarctions leading to considerable morbidity and mortality. During the past decade a group of molecules referred to as statins such as simvastatin, atrovastatin have been tried with great success in reducing total cholesterol. These molecules act by inhibiting the HMG CoA reductase enzyme thereby interfering with the synthesis of cholesterol. But statins reduce all the cholesterol including HDL cholesterol. Long term drug vigilance activity has revealed serious side effects of tendinopathy and related musculoskeletal disorders in some of the subjects. In an effort to manage hypercholesterolemia without serious side effects in a natural way we had tried the use of Amlamax a reconstituted, purified, standardized dried extract of amla (Emblica officinalis) containing 30% ellagitannins with other hydrolysable tannins on humans. We report the hitherto unobserved significant elevation of HDL cholesterol by the administration of Amlamax.

Locomotor activity rhythm in Drosophila melanogaster after 600 generations in an aperiodic environment.

The locomotor activity rhythm of flies from four populations of Drosophila melanogaster, maintained under constant light for more than 600 generations, was recorded in continuous light (LL) and continuous darkness (DD) using four different protocols. The main objective behind these experiments was to estimate the proportion of flies exhibiting circadian rhythm of locomotor activity in LL, and to investigate whether this could be increased by subjecting the flies to various light regimes. About 26% of the flies exhibited a circadian rhythm of locomotor activity in LL, and the proportion increased to about 48% after an exposure to 12 h of darkness. About 77% of the flies exhibited a circadian locomotor activity rhythm in DD. Persistence of circadian locomotor activity rhythm in a considerable proportion of these flies suggests an intrinsic adaptive value to possessing circadian rhythmicity, derived, perhaps, from the need to synchronise various processes within the organism.

Developmental plasticity of the locomotor activity rhythm of Drosophila melanogaster.

We used four replicate outbred populations of Drosophila melanogaster to investigate whether the light regimes experienced during the pre-adult (larval and pupal) and early adult stages influence the free-running period (tau(DD)) of the circadian locomotor activity rhythm of adult flies. In a series of two experiments four different populations of flies were raised from egg to eclosion in constant light (LL), in light/dark (LD) 12:12h cycle, and in constant darkness (DD). In the first experiment the adult male and female flies were directly transferred into DD and their locomotor activity was monitored, while in the second experiment the locomotor activity of the emerging adult flies was first assayed in LD 12:12h for 15 days and then in DD for another 15 days. The tau(DD) of the locomotor activity rhythm of flies that were raised in all the three light regimes, LL, LD 12:12h and in DD was significantly different from each other. The tau(DD) of the locomotor activity rhythm of the flies, which were raised in DD during their pre-adult stages, was significantly shorter than that of flies that were raised as pre-adults in LL regime, which in turn was significantly shorter than that of flies raised in LD 12:12h regime. This pattern was consistent across both the experiments. The results of our experiments serve to emphasise the fact that in order to draw meaningful inferences about circadian rhythm parameters in insects, adequate attention should be paid to control and specify the environment in which pre-adult rearing takes place. The pattern of pre-adult and early adult light regime effects that we see differs from that previously observed in studies of mutant strains of D. melanogaster, and therefore, also points to the potential importance of inter-strain differences in the response of circadian organisation to external influences.

Does the difference in the timing of eclosion of the fruit fly Drosophila melanogaster reflect differences in the circadian organization?

The eclosion rhythm of a laboratory population of Drosophila melanogaster was studied under 12h light, 12h dark (LD 12:12) cycles. Although most of the flies were found to eclose just after "lights on" in LD 12:12, termed within gate (WG) flies, a few flies were found to eclose nearly 10h after peak eclosion, termed outside gate (OG) flies. The circadian parameters of the clocks controlling oviposition rhythms in the WG and the OG flies were estimated to understand the cause of such differences in the timing of eclosion. The distribution of the fraction of individual flies exhibiting single, multiple, and no significant period in the WG flies was significantly different from distribution in the OG flies. Compared to the WG flies, more OG flies were found to exhibit oviposition rhythm with multiple periodicity, whereas more WG flies exhibited an oviposition rhythm with a single significant period. The fraction of flies with arrhythmic oviposition was similar in both the WG and the OG flies. Free-running period tau in constant darkness (DD) and the phase angle difference psi in LD 12:12 for the oviposition rhythm of WG and OG flies were significantly different. These results suggest that the differences in the time of eclosion between the flies eclosing within the gate and outside the gate of eclosion are probably due to differences in the circadian system controlling eclosion, which is reflected by the differences in their oviposition rhythm.

Persistence of oviposition rhythm in individuals of Drosophila melanogaster reared in an aperiodic environment for several hundred generations.

The oviposition rhythm of individual flies of Drosophila melanogaster from a population maintained in an aperiodic environment (with light, temperature, humidity, and other factors which could provide time cues, kept constant) for several hundred generations was assayed in constant light (LL), in light/dark (LD 12:12 hr) cycle, and in constant darkness (DD). More than 50% of the flies assayed exhibited rhythmicity in oviposition in all three light regimes. The results indicate that the phenomenon of egg laying is rhythmic in individual D. melanogaster females and is controlled by an endogenous time keeping mechanism. The persistence of the oviposition rhythm in a large proportion of individuals in the population after several hundred generations of rearing in a constant environment strengthens the view that possessing biological clocks may confer some intrinsic fitness advantage even to organisms living in aperiodic environments. J. Exp. Zool. 290:541-549, 2001.

Oxidative cyclisation of cinnamyl ethers mediated by CAN: a stereoselective synthesis of 3,4-trans disubstituted tetrahydrofuran derivatives.

The oxidative cyclisation of cinnamyl ethers mediated by cerium(IV) ammonium nitrate results in the stereospecific formation of 3,4-trans disubstituted tetrahydrofuran derivatives in moderate to good yields.

A case for multiple oscillators controlling different circadian rhythms in Drosophila melanogaster.

A population of the fruit fly Drosophila melanogaster was raised in periodic light/dark (LD) cycles of 12:12 h for about 35 generations. Eclosion, locomotor activity, and oviposition were found to be rhythmic in these flies, when assayed in constant laboratory conditions where the light intensity, temperature, humidity and other factors which could possibly act as time cue for these flies, were kept constant. These rhythms also entrained to a LD cycle of 12:12 h in the laboratory with each of them adopting a different temporal niche. The free-running periods (tau) of the eclosion, locomotor activity and oviposition rhythms were significantly different from each other. The peak of eclosion and the onset of locomotor activity occurred during the light phase of the LD cycle, whereas the peak of oviposition was found to occur during the dark phase of the LD cycle. Based on these results, we conclude that different circadian oscillators control the eclosion, locomotor activity and oviposition rhythms in the fruit fly D. melanogaster.

The effect of different light regimes on adult life span in Drosophila melanogaster is partly mediated through reproductive output.

The effects of different light regimes on the fitness of organisms have typically been studied using mean or median adult life span as the sole index of physiological well-being. It is, however, known that life span is inversely related to reproductive output in many species. Moreover, the effects of a given environmental treatment on life span can be due to effects on either age-independent mortality or the "rate of aging," or a combination of both. Drawing evolutionary inferences from the effects of light regime on mean or median adult life span alone is difficult and, at best, speculative. We examined the effects of constant light (LL), alternating light-dark cycles (LD 12:12 h), and constant darkness (DD) on the life span of reproducing and virgin flies in four populations of Drosophila melanogaster and also estimated lifetime fecundity in the three light regimes. The light regime effects on life span were further dissected by examining the age-independent mortality and the Gompertz rate of aging under the three light regimes. While mean adult life span of reproducing males and females and virgin females was significantly shorter in LL compared to LD 12:12 h and DD, life-time egg production was highest in LL. Life span of virgin males was not significantly affected by light regime. The rate of aging in reproducing females was higher in LL as compared to DD, whereas age-independent mortality was higher in DD. As reproductive output, especially early in life, is a far more significant contributor to fitness than is life span, our results suggest that the earlier reported deleterious effects of LL on fitness are partly an artifact of examining life span alone, without considering other components of adult fitness that trade off with life span. Our results suggest that detailed investigation of the effects of light regime on the physiological and behavioral processes that accompany reproduction is necessary to fully understand the effects of different light regimes on adult fitness in Drosophila.

Evolution of reduced pre-adult viability and larval growth rate in laboratory populations of Drosophila melanogaster selected for shorter development time.

Four large (n > 1000) populations of Drosophila melanogaster, derived from control populations maintained on a 3 week discrete generation cycle, were subjected to selection for fast development and early reproduction. Egg to eclosion survivorship and development time and dry weight at eclosion were monitored every 10 generations. Over 70 generations of selection, development time in the selected populations decreased by approximately 36 h relative to controls, a 20% decline. The difference in male and female development time was also reduced in the selected populations. Flies from the selected populations were increasingly lighter at eclosion than controls, with the reduction in dry weight at eclosion over 70 generations of selection being approximately 45% in males and 39% in females. Larval growth rate (dry weight at eclosion/development time) was also reduced in the selected lines over 70 generations, relative to controls, by approximately 32% in males and 24% in females. However, part of this relative reduction was due to an increase in growth rate of the controls populations, presumably an expression of adaptation to conditions in our laboratory. After 50 generations of selection had elapsed, a considerable and increasing pre-adult viability cost to faster development became apparent, with viability in the selected populations being about 22% less than that of controls at generation 70 of selection.

Persistence of eclosion rhythm in Drosophila melanogaster after 600 generations in an aperiodic environment.

The ubiquity of circadian rhythms suggests that they have an intrinsic adaptive value (Ouyang et al. 1998; Ronneberg and Foster 1997). Some experiments have shown that organisms have enhanced longevity, development time or growth rates when maintained in environments whose periodicity closely matches their endogenous period (Aschoff et al. 1971; Highkin and Hanson 1954; Hillman 1956; Pittendrigh and Minis 1972; Went 1960). So far there has been no experimental evidence to show that circadian rhythms per se (i.e. periodicity itself, as opposed to phasing properties of a rhythm) confer a fitness advantage. We show that the circadian eclosion rhythm persists in a population of the fruitfly Drosophila melanogaster maintained in constant conditions of light, temperature, and humidity for over 600 generations. The results suggest that even in the absence of any environmental cycle there exists some intrinsic fitness value of circadian rhythms.

Differential scanning calorimetric studies on the thermotropic phase transitions of dry and hydrated forms of N-acylethanolamines of even chainlengths.

N-acylethanolamines (NAEs) have attracted the attention of researchers in the last two decades due to their occurrence in biological membranes under conditions of stress as well as under normal conditions. Differential scanning calorimetric studies have been carried out on dry and hydrated samples of a homologous series of N-acylethanolamines containing saturated acyl chains of even number of carbon atoms (n = 8-20). In both cases a major sharp endothermic transition was observed which occurs at the melting point for the dry NAEs whereas for the hydrated samples it occurs at considerably lower temperatures. The enthalpies and entropies corresponding to this transition could be fitted, in each case, to a straight line suggesting that the transition enthalpy and transition entropy consist of a fixed component from the polar head group and the terminal methyl group, whereas the contribution of the methylene groups, (CH2)n, is linearly proportional to the number of carbon atoms in it. The contributions of each methylene unit to the transition enthalpy and transition entropy of NAEs were found to be deltaH(inc) = 0.82 (+/-0.02) and 0.96 (+/-0.06) kcal mol(-1), and deltaS(inc) = 2.01 (+/- 0.06) and 2.37 (+/-0.17) cal mol(-1) K(-1), respectively, for the dry and hydrated samples of NAEs, whereas the end contributions arising from the head group and the terminal methyl group were determined to be deltaH(o) = -0.10 (+/-0.26) and -0.52 (+/-0.82) kcal mol(-1) and deltaS(o) = 2.12 (+/-0.71) and 3.1 (+/-2.3) cal mol(-1) K(-1), respectively, for the dry and hydrated samples of NAEs. These results are relevant to an understanding of the thermodynamics of the phase properties of NAEs in membranes.