On the nature of the horny scales of the pangolin

The pangolin scale is a horny derivative of the epidermis. It is complex in structure and is divisible into three distinct regions. The dorsal plate forms approximately one-sixth of the scale thickness. It is composed of flattened solid keratinized cells without basophilic nuclear remnants. This region tends to fray easily. The dorsal plate contains bound phospholipids and sulphydryl groups but is weak in disulphide bonds.
The bulk of the scale is made up of the intermediate plate formed of less flattened cells without basophilic nuclei. This region is rich in disulphide bonds but contains no appreciable bound phospholipids or sulphydryl groups.
The ventral plate is only a few cells thick and is rich in bound phospholipids, which also occur in the underlying scale bed epidermis.
These three regions of the scale are formed from separate epidermal germinal areas which do not develop a granular layer. Keratohyalin granules are, however, formed in the epidermis between the scales.
It is suggested on the basis of histological structure and dishribution of chemical constituents that pangolin scales are probably homologous with primate nails.
Evidence against the views that they are homologous with reptilian scales or are derived from compressed hairs is presented.     

Increased synaptosomal dopamine content and brain concentration of paraquat produced by selective dithiocarbamates

Exposure to pesticides may be a risk factor for Parkinson's disease based on epidemiologic data in humans, animal models and in vitro studies. Different dithiocarbamate pesticides potentiate the toxicity of both 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine and paraquat in mouse models of Parkinsonism by an unknown mechanism. This study examined the effects of commercially used dithiocarbamates on [3H]dopamine transport in striatal synaptosomal vesicles and on the concentration of [14C]paraquat in vivo in mice. Different ethylenebis-dithiocarbamates and diethyl-dithiocarbamate increased dopamine accumulation in synaptosomes, whereas dimethyl-dithiocarbamate and methyl-dithiocarbamate did not. Increased dopamine accumulation in synaptosomes was dose dependent and was related to the carbon backbone of these molecules. The dithiocarbamates that increased accumulation of dopamine did not alter the influx of dopamine, but rather delayed the efflux out of synaptosomes. These same dithiocarbamates also increased the tissue content of [14C]paraquat in vivo by a mechanism that appeared to be distinct from the dopamine transporter. There was a consistent relationship between the dithiocarbamates that increased synaptosomal accumulation of dopamine and tissue content of paraquat, with those previously demonstrated to enhance paraquat toxicity in vivo. These results suggest that selective dithiocarbamates may alter the kinetics of different endogenous and exogenous compounds to enhance their neurotoxicity.     

The Behaviour of a Food Poisoning Strain of Clostridium welchii in Beef

S ummary : An inoculum of 10⁵ spores of Clostridium welchii F2985/50 in meat survived steaming at 100° for 5 h, the number being reduced sevenfold for every hour of steaming. They also survived for at least 6 months in frozen meat stored at -5° and -20°, whereas vegetative cells died more rapidly at -5° than at -20°. In beef stored for 13 days at 1°, 5°, 10° and 15° there was no multiplication but a slow destruction of vegetative cells, but there was little change in the spore count. Slow multiplication occurred at 20° but at 25° and 37° growth was rapid. Only about 3% of the spores germinated without prior heat shock, so the majority failed to germinate in raw meat stored at any temperature, but did so once the meat had been heated. In meat which had been heated and allowed to cool almost all of the spores had lost their heat resistance.
It was found that the minimal growth temperature was related to pH and medium, so that meat with a pH higher than that used in these experiments (pH 5°7–5°8) would probably have a lower minimal growth temperature for these organisms and would thus be more susceptible to spoilage.     

Role of Disulphide Bonds in the Resistance of Bacillus cereus Spores to Gamma Irradiation and Heat

S ummary . Spores of Bacillus cereus were treated with thioglycollic acid which ruptures at least 10–30% of the spore disulphide bonds by reducing them to thiol groups. The treated spores were still viable and were sensitive to lysozyme but remained as resistant to γ-irradiation and to heat as untreated spores. Neither treated nor untreated spores were sensitized to irradiation by reagents which block thiol groups. The results did not indicate that the high content of disulphide bonds in spore coat protein protects spores against inactivation by irradiation or heat.     

Evidence for lysosomal reduction of cystine residues.

Previously reported evidence for the existence of a thiol: protein disulphide oxidoreductase in rat liver lysosomes has been re-examined and ambiguous results obtained. However, incubation of purified rat liver lysosomes with ¹²⁵I-labelled insulin at pH 5.5 shows that cathepsin D and a thiol-dependent enzyme other than cathepsin B or L are important in its digestion. The latter enzyme is most probably a thiol: protein disulphide oxidoreductase.     

A histochemical study of keratinization in the domestic fowl (Gallus gallus)

The microanatomy of the epidermis of the domestic fowl is described and related to the distribution of various histochemical constituents involved in keratinization.
The avian horny layer over the back is composed of a loose network of structurally solid horny cells. This is in contrast to most mammalian epidermal horny cells in which structural keratin is found only in the peripheral cytoplasm, and the interior of the keratinocyte contains soluble products of cytolysis with possibly some free keratin filaments dispersed in the fluid material.
The avian tarsal epidermal horny scales show similarities to both the scales of lizards and snakes and to mammalian tail scales which appear to be homologous structures.
It is suggested that a thin layer of cells containing no detectable disulphide bonds, found in the tarsal scale region of the young chick, is probably mechanically weak and may function as a fission plane for sloughing of the horny layer. A specialized epidermis and thickened horny layer is developed in the fowl on the plantar underside of the toes, but this is quite different in structure from the mammalian plantar epidermis.
The overlapping of zones rich in ribonucleic acid (RNA) and bound cysteine (SH) in the growing feather suggests that protein synthesis and the preparatory stages to keratin disulphide bonding normally occur concurrently in feather formation. This is in contrast to the growing hair which has a region rich in RNA followed immediately before it becomes keratinized by a discrete keratogenous zone weak in RNA but rich in bound cysteine.     

When dinner is dangerous: toxic frogs elicit species-specific responses from a generalist snake predator

In arms races between predators and prey, some evolved tactics are unbeatable by the other player. For example, many types of prey are inedible because they have evolved chemical defenses. In this case, prey death removes any selective advantage of toxicity to the prey but not the selective advantage to a predator of being able to consume the prey. In the absence of effective selection for postmortem persistence of the toxicity then, some chemical defenses probably break down rapidly after prey death. If so, predators can overcome the toxic defense simply by waiting for that breakdown before consuming the prey. Floodplain death adders (Acanthophis praelongus) are highly venomous frog-eating elapid snakes native to northern Australia. Some of the frogs they eat are nontoxic (Litoria nasuta), others produce gluelike mucus when seized by a predator (Limnodynastes convexiusculus), and one species (Litoria dahlii) is dangerously toxic to snakes. Both the glue and the toxin degrade within about 20 min of prey death. Adders deal with these prey types in different and highly stereotyped ways: they consume nontoxic frogs directly but envenomate and release the other taxa, waiting until the chemical defense loses its potency before consuming the prey.     

Structural studies on the inactivation of gamma-glutamylcysteine synthetase by the disulphide analogues of radioprotective cysteamine derivatives. Effects of aminoalkyl and hydroxyalkyl chain length and beta beta-bis-dimethylation.

Disulphide compounds have been shown to inactivate gamma-glutamylcysteine synthetase, the rate-limiting enzyme for GSH synthesis. Such compounds bind to a cysteine residue at or near the glutamate-binding site of the enzyme. This phenomenon is thought to be responsible for the synergistic toxicity of the thiophosphate radio- and chemo-protective agent WR2721 and the oxygen-radical generator 6-hydroxydopamine (2,4,6-trihydroxyphenethylamine). 6-Hydroxy-dopamine enhances conversion of WR2721 into its disulphide metabolite NN'-bis-(3-aminopropyl)cystamine, which, in turn, paralyses the synthetase. In an effort to identify radio- and chemo-protective thiols and thiol derivatives that do not have this toxicity, we have begun to define the structure-activity relationship that governs inactivation of the enzyme by analogues of WR2721 disulphide. NN'-Bis(aminoalkyl)cystamines and bis(hydroxyalkyl)cystamines with an alkyl chain length of C5 or greater are not inactivators of the synthetase. That this is not due solely to the size of these compounds is shown by the potent inactivation of the enzyme by SAPH3 disulphide, an extremely bulky cystamine analogue. beta beta-Bis-dimethylation of the cystamine portion of the molecule also obviates inactivation. This is almost certainly due to steric interference with disulphide interchange. These findings may facilitate the safe adjunctive use of the thiol counterparts of such compounds with oxygen-radical-generating chemotherapeutic agents, and may shed light on the structure of the region of the synthetase adjacent to the glutamate-binding site.     

The action of nabam, metham-sodium and other sulphur compounds on Heterodera schachtii cysts

When used in tests against eggs of Heterodera schachtii, sodium ethylenebisdithiocarbamate (nabam) solutions break down to give a number of compounds, some being hatch-inhibiting and others hatch-stimulating. The compound mainly responsible for the hatch-stimulating activity of nabam solutions is ethylenethiuram monosulphide. Stored solutions of sodium ethylenebisdithiocarbamate lose activity because the ethylenethiuram monosulphide disappears. Zinc and manganese ethylenebisdithiocarbamates were more active hatching agents than equimolar mixtures of zinc or manganese sulphates with nabam. Hatch stimulation was least with a zinc sulphate/nabam mixture of mole ratio 1.5:1, in which the dithiocarbamate ion became largely replaced by the sparingly soluble zinc dithiocarbamate. Increasing the zinc concentration in the mixture increased hatching because of the hatching activity of the zinc ion. Nabam solutions containing manganese sulphate were inactivated because the manganese ion catalysed decomposition to the hatch-inhibiting carbon disulphide. Sodium N-methyldithiocarbamate (metham-sodium, Vapam), which readily decomposed in aqueous solution, was toxic to, and prevented the hatching of, H. schachtii. Methylisothiocyanate, the major decomposition product, was toxic to H. schachtii at concentrations greater than 0.1 mM.     

Viruses occurring in white clover (Trifolium repens L.) from permanent pastures in Britain

The uptake of captan from aqueous solution by conidia of Neurospora crassa can be markedly reduced by pretreatment of the cells with thiol reagents such as iodoacetic acid (IOA). The nature of this reaction has been investigated and it is shown that IOA largely reacts with the soluble thiol pool of the spores. Captan, however, reacts with both soluble and insoluble thiols and it is suggested that whilst the former is a detoxication process the latter may provide the key to its toxic action. Using glutathione as a model soluble thiol the molar ratio and pH dependence of the captan detoxication process has been determined and compared with the cellular reactions. Assay of ³⁵S-labelled captan has shown that captan is almost completely decomposed by the spores and that one-third of the accumulated captan is converted to sulphur compounds fixed to insoluble cell entities.     

On some commensal decapod crustaceans from Singapore (Palaemonidae and Porcellanidae)

Accounts are given of the structure and some aspects of the biology of three commensal crustaceans from Singapore. One of these represents a new genus of prawns of the subfamily Pontoniinae; the others are little known members of the family Porcellanidae.
It is suggested that the apparent rarity of these and other tropical commensal decapods is probably a spurious phenomenon resulting from inadequate collecting methods.
The functional taxonomy of these commensals is discussed and it is suggested that many of their structural peculiarities can be interpreted as adaptations to their mode of life.     

Engineering microbes to produce biofuels

The current biofuels landscape is chaotic. It is controlled by the rules imposed by economic forces and driven by the necessity of finding new sources of energy, particularly motor fuels. The need is bringing forth great creativity in uncovering new candidate fuel molecules that can be made via metabolic engineering. These next generation fuels include long-chain alcohols, terpenoid hydrocarbons, and diesel-length alkanes. Renewable fuels contain carbon derived from carbon dioxide. The carbon dioxide is derived directly by a photosynthetic fuel-producing organism(s) or via intermediary biomass polymers that were previously derived from carbon dioxide. To use the latter economically, biomass depolymerization processes must improve and this is a very active area of research. There are competitive approaches with some groups using enzyme based methods and others using chemical catalysts. With the former, feedstock and end-product toxicity loom as major problems. Advances chiefly rest on the ability to manipulate biological systems. Computational and modular construction approaches are key. For example, novel metabolic networks have been constructed to make long-chain alcohols and hydrocarbons that have superior fuel properties over ethanol. A particularly exciting approach is to implement a direct utilization of solar energy to make a usable fuel. A number of approaches use the components of current biological systems, but re-engineer them for more direct, efficient production of fuels.     

Ultrastructure of Trypanosoma conorhini in the Crithidial Phase*

SYNOPSIS. The ultrastructure of the crithidial phase of T. conorhini culture has been studied. The structure of the plasma membrane is not easy to make out; only at a few points in highly magnified picit is seen as a double osmiophilic membrane with an intermedilayer of low density. Sub-pellicular tubules are present also around the flagellar pocket. The nucleus usually has a single large nucleolus, but sometimes this may be double. The flagellum has the sual 2 central and 9 double peripheral fibers, the latter having lateral arms. The structure of the kinetoplast is similar to that of other rypanosomatids but the division of the organelle seems to be more complex than has been described: the central, DNA-bearing lamellae duplicate, forming a double transverse band; 1 of these bands probably migrates toward the side before invagination of the membrane completes the division. Mitochondria are long tubular structures with few cristae, disposed chiefly along the periphery of the cell. A communication between the kinetoplast and tubular mitochondria is very frequent. The endoplasmic reticulum is poorly developed, represented chiefly by smooth membranes surrounding vesicles, rough-surfaced membranes being scanty in most cells. Several inclusions may be found, some probably lipids, others being of unknown nature.     

Effect of oxidative stress on protein thiols in the blue mussel Mytilus edulis: proteomic identification of target proteins

Protein thiols are targets of oxidative stress. Their modification was analysed in gill extracts of the mussel Mytilus edulis, exposed to menadione. Diagonal gel electrophoresis revealed two clusters of carbonylated proteins involved in interchain disulphide linkages. Immunoblotting identified these as being associated with protein disulphide isomerase (PDI) and actin and this was confirmed by immunoprecipitation. Protein free thiols (-SH) were identified in 2-DE separations by labelling with 5-iodoacetamidofluorescein (IAF). Cysteines involved in disulphide bridges were identified by blocking free -SH with N-ethylmaleimide, reducing disulphides with DTT and IAF labelling. Several protein spots containing free thiols disappeared on exposure to menadione. Conversely, new protein spots containing disulphides appeared in response to menadione which may be protective against oxidative stress. In-gel tryptic digestion followed by LC/MS-MS and database searching identified some of the free thiol targets: PDI; hsp gp96; calreticulin; heavy metal binding protein. Tubulin, PDI, enolase and gelsolin contained new disulphide bridges in response to menadione. Our findings indicate a protein level response to oxidative stress principally involving PDI, chaperone-like and cytoskeletal proteins. Since many environmental pollutants cause oxidative stress, studies on PDI and structural proteins may be particularly relevant to understanding toxicity in this popular sentinel species.     

An electron microscopical study of the development of peroxisomes during formation and germination of ascospores in the methylotrophic yeast Hansenula polymorpha

Ascospore formation was studied in liquid cultures of the yeast Hansenula polymorpha, previously grown under conditions in which the synthesis of alcohol oxidase was repressed (glucose as growth substrate) or derepressed (methanol, glycerol and dihydroxyacetone as growth substrates and after growth on malt agar plates). In ascospores obtained from repressed cells, generally one small peroxisome was present. The organelle probably originated from the small peroxisome, originally present in the vegetative cells. They had no crystalline inclusions and cytochemical experiments indicated the presence of catalase, urate oxidase and amino acid oxidase activities in these organelles. In ascospores obtained from derepressed cells, generally 1–3 crystalline peroxisomes were observed. These organelles also originated from the peroxisomes originally present in the vegetative cells by means of fragmentation or division. They contained, in addition to the enzymes characteristic for peroxisomes in spores from repressed cells, also alcohol oxidase. The latter enzyme is probably responsible for the crystalline substructure of these peroxisomes.Peroxisomes had no apparent physiological function in the process of ascosporogenesis. A glyoxysomal function of the organelles during germination of the ascospores was also not observed. Germination of mature ascospores in media containing different sources of carbon and nitrogen showed that the function of the peroxisomes present in ascospores of Hansenula polymorpha is probably identical to that in vegetative haploid cells. They are involved in the oxidative metabolism of different carbon and nitrogen sources. Their enzyme profile is a reflection of that of peroxisomes of vegetative cells and their presence may enable the formation of cells which are optimally adapted to environmental conditions extant during spore germination.     

Toxicological evaluation of zidovudine and novel chalcogen derivatives in Drosophila melanogaster

Zidovudine (AZT) is the most commonly prescribed antiviral drug for the treatment of human immunodeficiency virus (HIV) infection. However, its chronic administration causes toxic side effects limiting its use. This study aimed to evaluate the toxicity of different concentrations of AZT and novel chalcogen derivatives (7A, 7D, 7G, 7K, 7M) on locomotion, mitochondrial dysfunction, acetylcholinesterase (AChE) activity, and production of reactive oxygen species (ROS) in adult Drosophila melanogaster. Our results show that AZT and its derivative 7K at a concentration of 10 μM impaired flies' locomotor behavior. Furthermore, AZT and the derivatives 7K, 7A, and 7M induced mitochondrial dysfunction observed by a decrease in oxygen flux through mitochondrial complexes I and II. Neither of the compounds tested affected AChE activity or ROS production in flies. According to these data, AZT derivatives presented the following decreasing order of toxicity: 7K > AZT > 7G > 7A > 7M > 7D. Based on the chemical structure, it is possible to infer that the presence of the seleno-phenyl group in 7A and 7G increases their toxicity compared to compounds 7D and 7M. In addition, compounds 7G, 7M, and 7K with three carbon atoms as spacer were more toxic than analogs containing one carbon atom (7A and 7D). Finally, the insertion of a p-methoxyl group enhances toxicity (7K). Based on these results, excepting 7K, all other chalcogen derivatives presented lower toxicity than AZT and are potential drug candidates.     

An Ultrastructural Study of Vairimorpha necatrix (Microspora, Microsporida) with Particular Reference to Episporontal Inclusions During Octosporogony

ABSTRACT. The life cycle of Vairimorpha necatrix was studied by electron microscopy. Disporous development has two distinct stages: 1) diplokaryotic meronts which are actively mitotic, and 2) diplokaryotic sporonts which are distinguished by reduced ribosome density and a thickened plasmalemma. After final division of the sporont, sporoblasts form spores which are ovocylindrical and measure 4.4 ± 0.08 × 2.3 ± 0.05 μ m (mean ± SE). Octosporous development results in eight haploid spores being formed in a sporophorous vesicle. The uninucleate octospores were smaller than the binucleate dispores and the exospore was thicker but less crenulate in outline. Early in octosporogony, tubules are produced from the sporont plasmalemma and electron-dense material accumulates in the episporontal space. The latter may be amorphous, vesiculated, or vacuolated in appearance and in later stages may take a stacked, lamellar form. At sporoblast formation, exospore material coats the plasmalemma and attached tubules; all inclusions in the episporontal space gradually disappear as spores are formed. These secretory products may have application to taxonomic distinction at the species level.     

Amino acid sequence of atrial natriuretic peptides in human coronary sinus plasma

Two atrial natriuretic peptides were purified from pooled human coronary sinus plasma by Sep-Pak extraction, immunoaffinity chromatography and reverse phase HPLC. The amino acid sequences of the two peptides were homologous with 99-126 human atrial natriuretic peptide (hANP) and 106-126 hANP, the latter being most probably linked to 99-105 ANP by the disulphide bond. The molar ratio of the peptides in plasma, as assessed by radioimmunoassay was 10:3.     

THE INSECTICIDAL ACTIVITY OF SUBSTANCES RELATED TO THE PYRETHRINS

The toxicity as contact insecticides of the esters of (±)-3-methyl-2-allyl-cycfopent-3-en-4-ol-i-one with the natural (+ )-trans and with synthetic (±) -cis-trans-chrysaii- themum monocarboxylic acids has been compared with that of the natural pyrethrins. The comparison was carried out on five species of test-insects.
It was found that the figure for both the absolute and the relative toxicities of these compounds varied with the species used as test-subject.
The factors involved in this variation are discussed. The extremes of variation of relative toxicity for the compounds with the natural acid were from about one-eighth as toxic as the pyrethrum standard when the aphid Macrosiphum solanifolii was used as test subject to nearly four times as toxic as the standard with the larvae of the moth Plutella maculipennis. The fully synthetic material varied from about one-sixteenth as toxic as the standard to Macrosiphum solanifolii to nearly twice as toxic to the larvae of Plutella maculipennis. The compound with the natural acid was approximately twice as toxic as the fully synthetic material to three of the insect species, but the fully synthetic material was considerably more than half as toxic as the partially synthetic to the fourth test-species. It is pointed out that while it is widely recognized that large differences in relative toxicity may occur when the effect of chemicals of widely different structure and mechanism of action are compared on a number of different test-species, the fact that these differences may also occur with related chemicals with, presumably, a similar mechanism of action, has not been clearly stated.
Even when the differences in relative toxicity are taken into account, the two synthetic pyrethrin-like esters still show high insecticidal activity.
It seems reasonable to suppose from the results that economic commercial synthesis of pyrethrin-like insecticides is not impossible.