Wax as a Mechanism for Protection against Photoinhibition — A Study of Cotyledon orbiculata

The leaves of the Crassulacean acid metabolism plant Cotyledon orbiculata have a waxy coating which is highly reflective but can be easily removed by brushing. This provided an ideal system in which to investigate the role of epidermal wax as a possible photoprotectant. Removal of the wax, prior to exposure to natural sunlight, resulted in substantial decreases in F_v/F_m and in severe cases evidence of photoinhibitory damage, as indicated by a rise in F_o. Leaves from which wax had been removed also showed higher conversion of violaxanthin to zeaxanthin than waxed leaves. Recovery of brushed leaves over a 12 day period was correlated with an increase in the total pool of xanthophyll cycle components. This study suggests that the presence of highly reflective wax on the epidermis may confer significant photoprotection to plants exposed to high solar radiation environments.     

Occurrence and Carbon Metabolism of Green Nonsulfur-like Bacteria in Californian and Nevada Hot Spring Microbial mats as Revealed by Wax Ester Lipid Analysis

Green nonsulfur-like bacteria (GNSLB) in Yellowstone hot spring microbial mats have been extensively studied and are thought to operate both as photoheterotrophs and photoautotrophs. Here we studied the occurrence and carbon metabolisms of GNSLB by analyzing the distribution and isotopic composition of their characteristic wax ester lipids in four Californian and Nevada hot spring microbial mats at a range of temperatures (37–96°C). The distribution of wax esters varied strongly with temperature. At temperatures between 50–60°C the wax ester composition in each of the four hot spring microbial mats was dominated by C₃₀ to C₃₆ wax esters, consisting of mixtures of C₁₅-C₁₈ n-alkyl and branched fatty acids and alcohols, typical for GNSLB. Stable carbon isotopic analysis showed that these wax esters were only depleted by 5 to 10‰ compared to dissolved inorganic carbon in the overlying water, suggesting that these GNSLB were mainly autotrophic. However, analysis of different depth layers of one microbial mat showed that these GNSLB wax esters were increasingly depleted in ¹³C with depth, suggesting that photoautotrophy mainly occurred in the top layer of the mat. ¹³C-depleted C₃₆-C₄₄ wax esters were found in one hot spring at high temperatures (77–96°C) and are likely derived from allochtonous plant waxes. At several lower temperature sites (35–40°C) the wax esters were predominantly composed of C₂₈, C₃₀ and C₃₂ wax esters consisting of mixtures of C₁₄-C₁₆ fatty acids and n-alkanols and were depleted in ¹³C by 15–20‰ relative to dissolved inorganic carbon, suggesting they may be derived from heterotrophic organisms. Our results indicate that autotrophic GNSLB occur widely in hot springs and that diverse groups of organisms contribute to the pool of wax ester lipids in hot spring environments.     

Trace Metal Concentrations in Marsh Profiles Under the Influence of an Emerging Delta (Atchafalaya River and Wax Lake Delta) Overlying a Several Thousand Year Old (Former)Mississippi River Delta Lobe

There are concerns over the increasing concentrations of trace metals being found in the environment. Deltas are essentially integrators of watershed contamination as they are the repositories of sediment transported from and through the watershed. In order to assess changes in trace metal concentrations transported by the Mississippi River–Atchafalaya River systems, vibracores were collected from three coastal freshwater marsh sites under the influence of the Atchafalaya River and emerging Wax Lake Delta (WLD). The cores extended to a depth of 4 m which included deposits of an earlier Mississippi River Delta lobe. C-14 dating showed an age at the lower depth corresponding to approximately 3500 years ago. Vertical profile distribution of metals and metalloids were measured and comparisons between older deposits and concentrations in recent deposits were made. Concentrations of As, Cr, Cu, Ni, Zn, Mo, V, Co, and Hg metals were measured in the profiles along with Fe, Mn, and Al. There was no significant increase in heavy metals or metalloids in recent years as compared to more than 3000-year-old sediment associated with an earlier Mississippi River Delta lobe. Results show that sediment diversion through the Wax Lake Outlet did not increase concentrations of these metals in surface marsh soils. The metal concentrations in the marsh profile were compared to ERL (effects range low) and ERM (effects range medium) values to allow an ecotoxicological assessment. Arsenic was below the ERM values but was greater than the ERL which indicate a potential toxicity under certain conditions. All other metal/metalloids measured were below ERL limits. This study suggests that planned Mississippi River sediment diversions designed to slow the rate of coastal land loss are not likely to lead to trace metals contamination.     

Plants as a Source of Bacterial Resistance Modulators and Anti-Infective Agents

The spread of multidrug-resistant (MDR) strains of bacteria necessitates the discovery of new classes of antibacterials and compounds that inhibit these resistance mechanisms. At present, there are no single chemical entity plant-derived antibacterials used clinically, and this chemically diverse group deserves consideration as a source for two major reasons. First, plants have exceptional ability to produce cytotoxic agents and second there is an ecological rationale that antimicrobial natural products should be present or synthesised de novo in plants following microbial attack to protect the producer from pathogenic microbes in its environment. We have been characterising plant-derived products that are either antibacterial in their own right, or modulators of resistance in bacterial strains possessing multidrug efflux mechanisms. These efflux transporters are responsible for resistance to certain antibiotics and antiseptics and occur in strains of methicillin-resistant Staphylococcus aureus (MRSA), a major clinical problem at present. We are also investigating plant sources for compounds with activity against mycobacteria with a view to discovering drug leads with potential activity toward tuberculosis (TB) producing species. This paper will briefly review the literature on plant derived bacterial resistance modifying agents and antibacterials. Examples in this area from our own work will be given. The activities of plant-derived antibacterials show that there are many potential new classes of antibacterial agents which should undergo further cytotoxicity, microbial specificity and preclinical studies.     

The Transgenic Rabbit as Model for Human Diseases and as a Source of Biologically Active Recombinant Proteins

Until recently, transgenic rabbits were produced exclusively by pronuclear microinjection which results in additive random insertional transgenesis; however, progress in somatic cell cloning based on nuclear transfer will soon make it possible to produce rabbits with modifications to specific genes by the combination of homologous recombination and subsequent prescreening of nuclear donor cells. Transgenic rabbits have been found to be excellent animal models for inherited and acquired human diseases including hypertrophic cardiomyopathy, perturbed lipoprotein metabolism and atherosclerosis. Transgenic rabbits have also proved to be suitable bioreactors for the production of recombinant protein both on an experimental and a commercial scale. This review summarizes recent research based on the transgenic rabbit model.     

Total synthesis and biological evaluation of viscolin, a 1,3-diphenylpropane as a novel potent anti-inflammatory agent

Total synthesis of viscolin, an anti-inflammatory 1,3-diphenylpropane isolated from Viscum coloratum, employing the Wittig reaction is reported. Key steps in the synthesis of viscolin depend on the selection of protecting groups to maintain the para hydroxyl group that is the most critical chemical structure influencing the biological activity of viscolin and the utilization of microwave-assisted Wittig olefination reaction. Anti-inflammatory potency of the synthetic viscolin, its precursor product 16, and its analogue 17, through their effects on reactive oxygen species (ROS), nitric oxide (NO), and pro-inflammatory cytokine production in leukocytes and microglial cells were evaluated. Excellent inhibition of ROS and NO production in inflammatory cells could confer the synthetic viscolin to be a potent anti-inflammatory agent for the treatment of oxidative stress-induced diseases.     

The importance of leaf surface wax as short-range attractant and oviposition stimulant in a generalist Lepidoptera

Green gram, Vigna radiata (L.) Wilczek, is an important pulse crop of Asia. Severe attack by the larvae of Spilosoma obliqua Walker (Lepidoptera: Arctiidae) causes defoliation of green gram and reduces seed yield. Females lay eggs on the leaf surface, and therefore, surface wax plays an important role as short-range attractant and oviposition stimulant. So, we have attempted to find out whether leaf surface wax compounds (alkanes and free fatty acids) from three green gram cultivars (PDM 54, PUSA BAISAKHI and SAMRAT) could act as short-range attractant and oviposition stimulant in females. The TLC, GC-MS and GC-FID analyses of n-hexane extracts revealed 20 n-alkanes from n-C₁₅ to n-C₃₆ and 13 free fatty acids from C12:0 to C21:0, whilst linoleic acid was unique in SAMRAT. Pentacosane was the predominant amongst n-alkanes in the leaf surface waxes of three cultivars. Heneicosanoic acid and palmitoleic acid were the predominant free fatty acids in the leaf surface waxes of PDM 54, and PUSA BAISAKHI and SAMRAT, respectively. Females were attracted towards one leaf equivalent surface wax of three green gram cultivars against solvent controls (n-hexane) in Y-tube olfactometer bioassays. A synthetic blend of pentacosane, heptacosane, nonacosane, hexatriacontane, palmitoleic acid, linolenic acid and stearic acid, a synthetic blend of pentacosane, hexatriacontane and stearic acid, and a synthetic blend of hexatriacontane, linolenic acid and stearic acid resembling in amounts present in one leaf equivalent surface wax of PDM 54, PUSA BAISAKHI and SAMRAT, respectively, served as short-range attractant and oviposition stimulant in females. Females showed equal preference for egg laying towards the above three synthetic blends when these blends were tested against each other, and hence, these blends could be employed in development of baited traps in pest management strategies.     

Red Algae of Peter the Great Bay as a Source of Arachidonic and Eicosapentaenoic Acids

In order to find new sources of arachidonic (AA) and eicosapentaenoic (EPA) acids, the composition of fatty acids was studied and lipid concentrations were determined in the thalluses of 32 species of red algae from Peter the Great Bay, Sea of Japan. The greatest level of EPA and a small concentration of AA were registered in the thalluses of Corallina pilulifera, Palmaria stenogona, Halosaccion yendoi, and Laurencia nipponica. Taking into consideration the level of the lipid concentrations in the algae, as well as their biomass and frequency of occurrence, the algae C. pilulifera, P. stenogona, L. nipponica, and Polysiphonia morrowii may be of interest as potential sources of EPA. Among the examined algae, only Gracilaria verrucosa showed a high level of AA.     

Cerebral Alanine Transport and Alanine Aminotransferase Reaction: Alanine as a Source of Neuronal Glutamate

Abstract: Alanine transport and the role of alanine amino-transferase in the synthesis and consumption of glutamate were investigated in the preparation of rat brain synaptosomes. Alanine was accumulated rapidly via both the high-and low-affinity uptake systems. The high-affinity transport was dependent on the sodium concentration gradient and membrane electrical potential, which suggests a cotransport with Na⁺. Rapid accumulation of the Na⁺-alanine complex by synaptosomes stimulated activity of the Na⁺/K⁺ pump and increased energy utilization; this, in turn, activated the ATP-producing pathways, glycolysis and oxidative phosphorylation. Accumulation of Na⁺ also caused a small depolarization of the plasma membrane, a rise in [Ca²⁺]₁, and a release of glutamate. Intra-synaptosomal metabolism of alanine via alanine aminotransferase, as estimated from measurements of N fluxes from labeled precursors, was much slower than the rate of alanine uptake, even in the presence of added oxoacids. The velocity of [¹⁵N]alanine formation from [¹⁵N]glutamine was seven to eight times higher than the rate of [¹⁵N]glutamate generation from [¹⁵N]alanine. It is concluded that (a) overloading of nerve endings with alanine could be deleterious to neuronal function because it increases release of glutamate; (b) the activity of synaptosomal alanine aminotransferase is much slower than that of glutaminase and hence unlikely to play a major role in maintaining [glutamate] during neuronal activity; and (c) alanine aminotransferase might serve as a source of glutamate during recovery from ischemia/hypoxia when the alanine concentration rises and that of glutamate falls.     

Sorbus aucuparia and Sorbus aria as a Source of Antioxidant Phenolics,Tocopherols, and Pigments

Due to its nutritive and medicinal properties, berries of some Sorbus species are used for the preparation of jams and jelly as well as in traditional medicine. On the other hand, their chemical composition is not much studied especially of those grown in Balkan Peninsula. We have analyzed individual phenolics, tocopherols, carotenoids and chlorophylls using HPLC in berries from Sorbus aucuparia and Sorbus aria collected in different localities in Serbia and Montenegro together with the amounts of total phenolics and proanthocyanidins as well as their radical scavenging activity against DPPH radical. Berries of S. aucuparia were richer source of polyphenolics in comparision with S. aria and, regardless the species and locality, caffeoylquinic acids such as neochlorogenic and chlorogenic acid were the most abundant compounds. Among analyzed tocopherols the most abundant in all samples was α‐tocopherol (0.48 – 19.85 μg/g dw) as it was β‐carotene among carotenoids (mean concentration of 0.98 μg/g dw in S. aucuparia and 0.40 μg/g dw in S. aria, respectively). Correlation between total phenolics and DPPH radical scavenging activity was noticed. Our study represents comprehensive report on chemical composition of S. aucuparia and S. aria which could contribute to a better understanding of their quality.     

Identification of the border between fibronectin type III homologous repeats 2 and 3 of the neural cell adhesion molecule L1 as a neurite outgrowth promoting and signal transducing domain

To determine the domains of the neural cell adhesion molecule L1 involved in neurite outgrowth, we have generated monoclonal antibodies against L1 and investigated their effects on neurite outgrowth of small cerebellar neurons in culture. When the 10 antibodies were coated as substrate, only antibody 557.B6, which recognizes an epitope represented by a synthetic peptide comprising amino acids 818 to 832 at the border between the fibronectin type III homologous repeats 2 and 3, was as efficacious as L1 in promoting neurite outgrowth, increasing intracellular levels of Ca²⁺, and stimulating the turnover of inositol phosphates. These findings suggest that neurite outgrowth and changes in these second messengers are correlated. Such a correlation was confirmed by the ability of Ca²⁺ channel antagonists and pertussis toxin to inhibit neurite outgrowth on L1 and antibody 557.B6. These observations indicate for the first time a distinct site on cell surface-bound-L1 as a prominent signal-transducing domain through which the recognition events appear to be funneled to trigger neurite outgrowth, increase turnover of inositol phosphates, and elevate intracellular levels of Ca²⁺. © 1995 John Wiley & Sons, Inc.     

Cassava as a Cheap Source of Carbon for Rhizobial Inoculant Production using an Amylase-producing Fungus and a Glycerol-producing Yeast

Summary The aim of this research was to develop methods to use low-cost carbon compounds for rhizobial inoculant production. Five raw starch materials; steamed cassava, sticky rice, fresh corn, dry corn and sorghum were tested for sugar production by an amylase-producing fungus. Streamed cassava produced the highest amount of reducing sugar after fermentation. Bradyrhizobium japonicum USDA110, Azorhizobium caulinodans IRBG23, Rhizobium phaseoli TAL1383, Sinorhizobium fredii HH103, and Mesorhizobium ciceri USDA2429 were tested on minimal medium supplemented with reducing sugar obtained from cassava fermentation. All strains, except B. japonicum USDA110, could grow in medium containing cassava sugar derived from 100 g steamed cassava per litre, and the growth rates for these strains were similar to those in medium containing 0.5 (w/v) mannitol. The sugar derived from steamed cassava was further used for production of glycerol using yeast. After 1 day of yeast fermentation, the culture containing glycerol and heat-killed yeast cells, was used to formulate media for culturing bradyrhizobia. A formulation medium, FM4, with a glycerol concentration of 0.6 g/l and yeast cells (OD₆₀₀ = 0.1) supported growth of B. japonicum USDA110 up to 3.61 × 10⁹ c.f.u./ml in 7 days. These results demonstrate that steamed cassava could be used to provide cheap and effective carbon sources for rhizobial inoculant production.     

The Effects of Using Trehalose as a Carbon Source on the Proliferation of Phalaenopsis and Doritaenopsis Protocorm-Like-Bodies

In this communication, the effects of trehalose and culture system on protocorm-like body (PLB) growth were investigated. PLB derived from Phalaenopsis and Doritaenopsis cultivars, which were grown on solidified trehalose amended NP medium showed higher proliferation rate than on NP medium containing sucrose. For P. ‘Hwa Feng Red Jewel’ and Dtp. ‘Mount Beauty×Su’s Red Lip’, the proliferation rates on solidified trehalose media were almost two times higher than those on sucrose media after 8-week culture. However, Knudson C (KC) medium did not reveal the similar results between trehalose and sucrose. In liquid culture system, both trehalose amended NP and KC media brought about better results than sucrose containing media for PLB proliferation. For culture system test, solidified media showed higher proliferation rate than liquid media under the same medium composition. Roller bottles were more suitable than flask-shaking cultures in liquid systems for PLB proliferation.     

The development of 1,3-diphenylisobenzofuran as a highly selective probe for the detection and quantitative determination of hydrogen peroxide

1,3-Diphenylisobenzofuran (DPBF) has been developed as a selective probe for the detection and quantitative determination of hydrogen peroxide in samples containing different reactive nitrogen and oxygen species (RNOS). DPBF is a fluorescent probe which, for almost 20 years, was believed to react in a highly specific manner toward some reactive oxygen species (ROS) such as singlet oxygen and hydroxy, alkyloxy or alkylperoxy radicals. Under the action of these individuals DPBF has been rapidly transformed to 1,2-dibenzoylbenzene (DBB). In order to check if DPBF can act as a unique indicator of the total amount of different RNOS, as well as oxidative stress caused by an overproduction of these individuals, a series of experiments was carried out, in which DPBF reacted with peroxynitrite anion, superoxide anion, hydrogen peroxide, hypochlorite anion, and anions commonly present under biological conditions, namely nitrite and nitrate. In all cases, except for hydrogen peroxide, the product of the reaction is DBB. Only under the action of H₂O₂ 9-hydroxyanthracen-10(9H)-one (oxanthrone) is formed. This product has been identified with the use of fluorescence spectroscopy, NMR spectroscopy, high performance liquid chromatography coupled with mass spectrometry, infrared spectroscopy, elemental analysis, and cyclic voltammetry (CV). A linear relationship was found between a decrease in the fluorescence intensity of DPBF and the concentration of hydrogen peroxide in the range of concentrations of 0.196–3.941?mM. DPBF responds to hydrogen peroxide in a very specific way with the limits of detection and quantitation of 88 and 122.8?μM, respectively. The kinetics of the reaction between DBBF and H₂O₂ was also studied.     

Surfacing activity and food utilisation in the obligatory air-breathing fish ophiocephalus striatus as a function of body weight

Reared in cylindrical aquaria containing different depths of water (2.5 to 70 cm) the obligatory air-breathing fish Ophiocephalus striatus, belonging to different weight classes (0.1, 0.75, 10, 20 and 41 g), was forced to swim vertically a longer or shorter distance per surfacing. Surfacing frequency was a depth-dependent, activity in individuals weighing less than 20 g in all weight classes, the frequency was nearly 2 times more in the series fed ad libitum on fish muscle, than in the one where the fish were starved. Owing to the sustained surfacing activity and the consequent fatigue, the test individuals hung to the surface for a definite period. Neither frequency nor duration of hanging was depth-dependent. Mean hanging durations for the feeding series were 1.1, 3.8, 5.9, 7.9 and 8.8 hr/day in the 0.1, 0.75, 10, 20 and 41 g weight classes, respectively; the corresponding values for the starving series were 1.3, 15.0, 13.0, 12.7 and 12.8 hr/day. The distance swum by the feeding fish increased from 57 to 681 m/day and from 61 to 507 m/day in the 0.1 and 41 g individuals exposed to the minimum and maximum aquarium depths. Feeding rate, which was a depth-dependent activity, decreased from 280 to 113 g cal/g live fish/day with increasing weight. Rate and efficiency of conversion also decreased with increasing body weight; in larger fish conversion was dependent on volume rather than on depth of the aquarium. O₂ uptake of feeding fish was about 6 times higher than the starving ones of the tested weight classes at different aquarium depths.This paper is part of a thesis submitted by the author (from A.P.A. College, PALNI) to Madurai University in partial fulfillment of the requirement of the Ph. D. degree. Contribution No. 12 under a research scheme granted to Dr. T. J. Pandian by the UGC (New Delhi). The author is grateful to Dr. T. J. Pandian for valuable guidance, financial support and encouragements.     

Fetal and Newborn Calf Thymus as a Source of Chromatin Proteins: Purification of HMG-1 and HMG-2

The high mobility group (HMG) non-histone chromosomal proteins were first isolated from calf thymus' but were later found in numerous organs of many vertebrates.' The proteins can be extracted from calf thymus 1 with 0.35 M NaCl and they are quite soluble in 2% trichloroacetic acid. We have shown that members of the HMG-1 family (i.e., HMG-1, HMG-2, and HMG-E) exhibit a preferential affinity for single-stranded DNA at roughly physiological ionic trength. Members of this family have other intriguing properties (see references 6 and 7 for recent reviews), including the ability to assemble nucleosomes in vitroe8 The architecture of the proteins strongly suggests that they are designed to interact simultaneously with histones and with DNA through physically distinct domains^{6, 9}.     

Methane as an Organic Matter Source and the Trophic Basis of a Laptev Sea Cold Seep Microbial Community

An area of cold methane seeps at the bottom of the Laptev Sea was investigated. High rates of methane oxidation were revealed in the sediments and in the water column. Anaerobic methane oxidation carried out by the ANME-2 a/b consortium was coupled to sulfate reduction. Bacteria of the genera Sulfurovum and Arcobacter were the agents of the sulfur cycle. Methane unconsumed in the sediments diffused into the near-bottom water, where it was oxidized by methanotrophic bacteria. Methanotrophic activity was essential for development of symbiotrophic tubeworms of the upper sediment layers, which were responsible for the process of bioturbation.     

Balkans’ Asteraceae Species as a Source of Biologically Active Compounds for the Pharmaceutical and Food Industry

Herbal drugs are a useful source of different bioactive compounds. Asteraceae species, as the most widespread vascular plants, can be used both as food and as medicine due to the great diversity of recorded chemical components – different phenolic compounds, terpenes, carotenoids, vitamins, alkaloids, etc. The Balkan Peninsula is characterized by great diversity of plants from Asteraceae family, including presence of rare and endemic species. In this review, results of the survey of chemical composition and biological activity, mainly focusing on antioxidant, antimicrobial and anticancer effects of selected Balkans’ Asteraceae species were provided. In addition, information on edible plants from Asteraceae family is presented, due to growing interest for the so‐called ‘healthy diet’ and possible application of Balkans’ Asteraceae species as food of high nutritional value or as a source of functional food ingredients.