Thyroid function in a lizard, a tortoise and a crocodile, compared with mammals

1. Thyroid activity was examined in the lizard, Trachydosaurus rugosus, the tortoise Chelodina longicollis and the crocodile, Crocodylus johnstoni, acclimated to 20-22 degrees C and 30-32 degrees C. Thyroidal uptake and release of 125I, plasma concentrations of T3 and T4 were measured as was resting oxygen consumption (at 30 degrees C) before and after both thyroidectomy and thyroxine injections. 2. All three species showed 125I uptake at both temperatures and showed no thyroidal release of 125I at 20-22 degrees C but exhibited thyroidal release of 125I (and presumably hormone secretion) at 30-32 degrees C. 3. Plasma concentrations of thyroxine ranged from 0.55 nM to 3.24 nM and triiodothyronine from 0.14 nM to 0.51 nM. 4. Neither thyroidectomy nor thyroxine injections had any effect on metabolic rate in 20-22 degrees C acclimated lizards. Thyroidectomy resulted in a significant decrease in metabolic rate in 30-32 degrees C acclimated lizards and tortoises and thyroxine injections resulted in significant increases in metabolism in 30-32 degrees C acclimated lizards, tortoises and crocodiles. 5. A comparison of thyroid parameters in reptiles and mammals concluded that although the reptilian thyroid is active at high temperatures it is still considerably less active than it is in mammals.     

Extensive pollen dispersal in a bird-pollinated shrub, Calothamnus quadrifidus, in a fragmented landscape

Pollen dispersal was investigated in six populations of Calothamnus quadrifidus, a bird-pollinated shrub in the fragmented agricultural region of southern Western Australia. Paternity analysis using six microsatellite loci identified a pollen source within populations for 67% of seedlings, and the remainder were assumed to have arisen from pollen sources outside the populations. Outcrossing was variable, ranging from 5% to 82%, and long-distance pollen dispersal was observed in all populations with up to 43% of pollen sourced from outside the populations over distances of up to 5 km. This extensive pollen immigration was positively associated with population size but not isolation. Comparison of two populations of similar size but different density showed greater internal pollination and less selfing in the denser population, suggesting an influence of density on pollinator behaviour. The study revealed extensive long-distance pollen dispersal for C. quadrifidus within this fragmented agricultural landscape and highlighted the interaction between reserve populations and isolated road verge remnants in maintaining genetic connectivity at the landscape scale.     

Production of a high viscosity polysaccharide, methylan, in a novel bioreactor

The effect of shear stress on the production of a high viscosity polysaccharide, methylan, from methanol by Methylobacterium organophilum was investigated by using a multidisk mixer. It was observed in the multidisk mixer with defined shear stresses that the specific production rate of methylan increased gradually with increasing shear stress up to 30 Pa, and the production rate was constant beyond 30 Pa. This result suggested that the limited mass transfer from the medium into cells reduced methylan production. A novel bioreactor that provided the large volume of a high shear region was used to increase methylan production. Fed-batch cultures in the novel bioreactor were performed by the dissolved oxygen-stat method of methanol. When 1.13 g/L ammonium ion was added, the concentrations of cells of methylan were 31 and 20.6 g/L, respectively. The productions of cells and methylan in our designed bioreactor were 20 and 50% higher than those obtained in a conventional fermentor. The methylan content reached a maximum of 20.7 g/L in the bioreactor and the viscosity of the fermentation broth was 127 Pa . s, which corresponds to 68 g/L as a xanthan. (c) 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 54: 115-121, 1997.     

Adelmidrol, a palmitoylethanolamide analogue, reduces chronic inflammation in a carrageenin-granuloma model in rats

Palmitoylethanolamide (PEA) and some of its analogues have shown great efficacy in the treatment of pain and inflammation. Adelmidrol – the International Nonproprietary Name (INN) of the di-amide derivative of azelaic acid – is one of these analogues. The anti-inflammatory and analgesic effects of PEA and adelmidrol are hypothesized to be mediated, at least in part, by mast cell down-modulation. Mast cell mediators released at early stage of the inflammatory process drive the inflammatory reaction to chronicity as it happens in X-carrageenin-induced granulomatous tissue formation. In the present study, the choice of testing adelmidrol depends upon the physicochemical properties of the compound, i.e. the amphipatic feature, that make it more easily soluble than PEA. In this study, we investigated the effect of adelmidrol on granuloma formation induced by λ-carrageenin-soaked sponge implant in rats. Our results show that the local administration of the compound under study significantly decreases weight and neo-angiogenesis in granulomatous tissue. The anti-inflammatory effect was due to the modulation of mast cells degranulation, as shown by histological analysis and by the inhibition of the release of several pro-inflammatory and pro-angiogenic enzymes ( e.g. iNOS, chymase and metalloproteinase MMP-9), and mediators ( e.g. nitric oxide and TNF-α). The results indicate that adelmidrol, given locally, may represent a potential therapeutic tool in controlling chronic inflammation.     

Teloplasm formation in a leech, Helobdella triserialis, is a microtubule-dependent process

Fertilized eggs of the leech Helobdella triserialis undergo a cytoplasmic reorganization which generates domains of nonyolky cytoplasm, called teloplasm, at the animal and vegetal poles. The segregation of teloplasm to one cell of the eight-cell embryo is responsible for a unique developmental fate of that cell, i.e., to give rise to segmental ectoderm and mesoderm. We have studied the cytoplasmic movements that generate teloplasm using time-lapse video microscopy; the formation and migration of rings of nonyolky cytoplasm were visualized using transmitted light, while the movements of mitochondria into these rings were monitored with epifluorescence after labeling embryos with rhodamine 123, a fluorescent mitochondrial dye. To examine the likelihood that cytoskeletal elements play a role in the mechanism of teloplasm formation in Helobdella, we examined the distribution of microtubules and microfilaments during the first cell cycle by indirect immunofluorescence and rhodamine-phalloidin labeling, respectively. The cortex of the early embryo contained a network of microtubules many of which were oriented parallel to the cell surface. As teloplasm formation ensued, microtubule networks became concentrated in the animal and the vegetal cortex relative to the equatorial cortex. More extensive microtubule arrays were found within the rings of teloplasm. Actin filaments appeared in the form of narrow rings in the cortex, but these varied apparently randomly from embryo to embryo in terms of number, size, and position. The role of microtubules and microfilaments in teloplasm formation was tested using depolymerizing agents. Teloplasm formation was blocked by microtubule inhibitors, but not by microfilament inhibitors. These results differ significantly from those obtained in embryos of the oligochaete Tubifex hattai, suggesting that the presumably homologous cytoplasmic reorganizations seen in these two annelids have different cytoskeletal dependencies.     

Excitation energy relaxation in a symbiotic cyanobacterium, Prochloron didemni, occurring in coral-reef ascidians, and in a free-living cyanobacterium, Prochlorothrix hollandica

The marine cyanobacterium Prochloron is a unique photosynthetic organism that lives in obligate symbiosis with colonial ascidians. We compared Prochloron harbored in four different host species and cultured Prochlorothrix by means of spectroscopic measurements, including time-resolved fluorescence, to investigate host-induced differences in light-harvesting strategies between the cyanobacteria. The light-harvesting efficiency of photosystems including antenna Pcb, PS II-PS I connection, and pigment status, especially that of PS I Red Chls, were different among the four samples. We also discuss relationships between these observed characteristics and the light conditions, to which Prochloron cells are exposed, influenced by distribution pattern in the host colonies, presence or absence of tunic spicules, and microenvironments within the ascidians' habitat.     

An allotypic specificity presumably of the a series in rabbit immunoglobulins, different from a1, a2, and a3

From the serum of a wild rabbit lacking all the known allotypic specificities of the a series, IgG showing an allotypic specificity named. A100 has been isolated and antisera against it prepared in domestic rabbits. The determinants responsible for the A100 allotypic specificity are present both on IgG and IgM. They are located on the heavy chain and the Fab fragment of IgG.Evidence for the genetic determinism of A100 suggests that it is the product of a new allele at the $\text{a}$ locus.     

Spargana in a Weasel,Mustela sibirica manchurica,and a Wild Boar,Sus scrofa,from Gangwon-do,Korea

To know the status of sparganum (plerocercoid of Spirometra erinacei) infection in the Korean wild life, several species of wild animals were captured in Gangwon-do and examined for their status of infection with spargana. From February to December 2011, a total of 62 wild boars, 5 badgers, 1 weasel, 1 Siberian chipmunk, and 53 wild rodents were captured, and their whole muscles were examined with naked eyes for the presence of spargana worms. From the weasel and 1 wild boar, a total of 5 spargana specimens were extracted. The weasel was for the first time recorded as an intermediate or paratenic/transport host of S. erinacei in Korea, and both the weasel (Mustela sibirica manchurica) and wild boar (Sus scrofa) were added to the list of wild animals carrying spargana.     

Interactions between a braconid,Microplitis croceipes,and a fungus,Nomuraea rileyi,in laboratory-reared bollworm larvae

The parasite Microplitis croceipes required 1.1 days longer at 26°C to complete development in Heliothis zea larvae than was required for the fungus Nomuraea rileyi to kill the host larvae and sporulate. Host larvae parasitized by M. croceipes or infected with N. rileyi failed to complete a fifth larval molt or pupate. Of the remaining healthy larvae, one-half completed six larval stadia before popation. Larvae parasitized by M. croceipes were predisposed to infection by N. rileyi, but the fungus inhibited development of M. croceipes if host larvae were infected with N. rileyi within 1 day after parasitization.     

CARPEL FACTORY,a Dicer homolog,and HEN1, a novel protein,act in microRNA metabolism in Arabidopsis thaliana

BACKGROUND: In metazoans, microRNAs, or miRNAs, constitute a growing family of small regulatory RNAs that are usually 19-25 nucleotides in length. They are processed from longer precursor RNAs that fold into stem-loop structures by the ribonuclease Dicer and are thought to regulate gene expression by base pairing with RNAs of protein-coding genes. In Arabidopsis thaliana, mutations in CARPEL FACTORY (CAF), a Dicer homolog, and those in a novel gene, HEN1, result in similar, multifaceted developmental defects, suggesting a similar function of the two genes, possibly in miRNA metabolism.RESULTS: To investigate the potential functions of CAF and HEN1 in miRNA metabolism, we aimed to isolate miRNAs from Arabidopsis and examine their accumulation during plant development in wild-type plants and in hen1-1 and caf-1 mutant plants. We have isolated 11 miRNAs, some of which have potential homologs in tobacco, rice, and maize. The putative precursors of these miRNAs have the capacity to form stable stem-loop structures. The accumulation of these miRNAs appears to be spatially or temporally controlled in plant development, and their abundance is greatly reduced in caf-1 and hen1-1 mutants. HEN1 homologs are found in bacterial, fungal, and metazoan genomes.CONCLUSIONS: miRNAs are present in both plant and animal kingdoms. An evolutionarily conserved mechanism involving a protein, known as Dicer in animals and CAF in Arabidopsis, operates in miRNA metabolism. HEN1 is a new player in miRNA accumulation in Arabidopsis, and HEN1 homologs in metazoans may have a similar function. The developmental defects associated with caf-1 and hen1-1 mutations and the patterns of miRNA accumulation suggest that miRNAs play fundamental roles in plant development.     

Protein W, a spore-specific protein in Myxococcus xanthus, formation of a large electron-dense particle in a spore

The gene for the major spore-specific protein, termed protein W, was cloned, and it was found that protein W is composed of 426 amino acid residues including 31% charged (133 residues) and 39% hydrophobic (166 residues) amino acids. In the protein, a motif consisting of five amino acid residues [(V/L/I)–R–E–R–(V/L/I)] is repeated 28 times, and another motif [M–M–(P/G)–Q–G] five times. Protein W is synthesized during a very late stage of development, forming a single, large electron-dense particle (200–400 nm in diameter) inside a spore. X-ray microanalysis of the particle revealed that it contains a high amount of phosphate in addition to calcium and magnesium. It is proposed that protein W consisting of highly charged repetitive sequences is a polyphosphate storage protein to store energy in spores. The disruption of the gene for protein W resulted in delayed fruiting body formation and a lower spore yield.     

Production,in Pichia pastoris,of a recombinant monomeric mapacalcine,a protein with anti-ischemic properties

Mapacalcine is a small homodimeric protein of 19 kDa with 9 disulfide bridges extracted from the Cliona vastifica sponge (Red Sea). It selectively blocks a calcium current insensitive to most calcium blockers. Specific receptors for mapacalcine have been described in a variety of tissues such as brain, smooth muscle, liver, and kidney. Previous works achieved on hepatocytes and nervous cells demonstrated that this protein selectively blocks a calcium influx triggered by an ischemia/reperfusion (I/R) shock and efficiently protects cells from death after I/R. The aim of this work was to produce the recombinant mapacalcine in the yeast Pichia pastoris. Mass spectrometry, light scattering analysis and biological characterization demonstrated that the recombinant mapacalcine obtained was a monomeric form with 4 disulfide bridges which retains the biological activity of the natural protein.     

Glucuronyl glycine,a novel N-terminus in a glycoprotein

Treatment of cutinase, an extracellular glycoprotein produced by Fusarium solani f. pisi, with NaB³H₄ at pH 7.0 generated labeled enzyme. Acid hydrolysis showed that all of the label was in an acidic carbohydrate which was identified as gulonic acid. The N-terminal amino group of the enzyme is blocked; the precursor of gulonic acid has a free reducing group and it is attached via a linkage resistant to β-elimination. Furthermore, pronase digestion of NaB³H₄-treated cutinase gave rise to a ninhydrin negative compound which contained the bulk of the ³H and this compound was identified as N-gulonyl glycine. These results strongly suggest that the amino group of glycine, the N-terminal amino acid of this enzyme, is in amide linkage with glucuronic acid.     

Granzyme a, a stealth killer in the mitochondrion

The induction of caspase-independent cell death by killer lymphocytes involves the serine protease granzyme A (GzmA). In this issue, Martinvalet et al. (2008) show that GzmA penetrates the mitochondrial matrix without perturbing normal mitochondrial functions. In the mitochondrial matrix, GzmA cleaves NDUFS3 (a component of the electron transport chain) leading to production of reactive oxygen species and ultimately to cell death.     

Phenotype of a mechanosensitive channel mutant, mid-1, in a filamentous fungus, Neurospora crassa

In the yeast Saccharomyces cerevisiae, the MID1 (mating-induced death) gene encodes a stretch-activated channel which is required for successful mating; the mutant phenotype is rescued by elevated extracellular calcium. Homologs of the MID1 gene are found in fungi that are morphologically complex compared to yeast, both Basidiomycetes and Ascomycetes. We explored the phenotype of a mid-1 knockout mutant in the filamentous ascomycete Neurospora crassa. The mutant exhibits lower growth vigor than the wild type (which is not rescued by replete calcium) and mates successfully. Thus, the role of the MID-1 protein differs from that of the homologous gene product in yeast. Hyphal cytology, growth on diverse carbon sources, turgor regulation, and circadian rhythms of the mid-1 mutant are all similar to those of the wild type. However, basal turgor is lower than wild type, as is the activity of the plasma membrane H(+)-ATPase (measured by cyanide [CN(-)]-induced depolarization of the energy-dependent component of the membrane potential). In addition, the mutant is unable to grow at low extracellular Ca(2+) levels or when cytoplasmic Ca(2+) is elevated with the Ca(2+) ionophore A23187. We conclude that the MID-1 protein plays a role in regulation of ion transport via Ca(2+) homeostasis and signaling. In the absence of normal ion transport activity, the mutant exhibits poorer growth.     

Hemps, a novel EGF-like protein, plays a central role in ascidian metamorphosis

All chordates share several characteristic features including a dorsal hollow neural tube, a notochord, a pharynx and an endostyle. Unlike other chordate taxa, ascidians have a biphasic life-history with two distinct body plans. During metamorphosis, the larval nerve cord and notochord degenerate and the pharyngeal gill slits and endostyle form. While ascidians, like other marine invertebrates, metamorphose in response to specific environmental cues, it remains unclear how these cues trigger metamorphosis. We have identified a novel gene (Hemps) which encodes a protein with a putative secretion signal sequence and four epidermal growth factor (EGF)-like repeats which is a key regulator of metamorphosis in the ascidian Herdmania curvata. Expression of Hemps increases markedly when the swimming tadpole larva becomes competent to undergo metamorphosis and then during the first 24 hours of metamorphosis. The Hemps protein is localised to the larval papillae and anterior epidermis of the larva in the region known to be required for metamorphosis. When the larva contacts an inductive cue the protein is released, spreading posteriorly and into the tunic as metamorphosis progresses. Metamorphosis is blocked by incubating larvae in anti-Hemps antibodies prior to the addition of the cue. Addition of recombinant Hemps protein to competent larvae induces metamorphosis in a concentration-dependent manner. A subgroup of genes are specifically induced during this process. These results demonstrate that the Hemps protein is a key regulator of ascidian metamorphosis and is distinct from previously described inducers of this process in terrestrial arthropods and aquatic vertebrates.     

PEP A9, a new,unstable variant in the peptidase a system

Summary A rare peptidase A variant, tentatively designated PEP A⁹, was observed in six members of a German family, indicating autosomal codominant inheritance. The electrophoretic mobility is similar to that of PEP A 3-1, but it has very low in vivo stability. There is no apparent association with a disease state. A simple and sensitive staining reagent for PEP A was found in o-phthalaldehyde.