bluuebunny/crossref_metadata_2025 huggingface dataset:
Relapsing polychondritis
Authors: Liu Clive M. , Hata Tissa R. , Swinyer Leonard , Petersen Marta J.
Published: September 2003

A 43‐year‐old Chilean man presented with a 5‐month history of progressive hypertrophy of the ears bilaterally. He was seen initially by a dermatologist in Chile for complaints of erythema and swelling of the ears, and had been treated unsuccessfully with topical steroids and antimicrobial ointments. On presentation to our clinic, the hypertrophy had stabilized and the erythema had resolved, but he complained of decreased hearing due to narrowing of the external auditory canal. Associated symptoms included occasional pruritus, but he denied any pain. He also denied a history of sinus problems, respiratory symptoms, ocular pain, chest pain, and arthralgias. Physical examination revealed firm hypertrophy of the collagenous areas of both ears, sparing the ear lobes (Fig. 1). No pain was elicited on palpation. No conjunctivitis was noted and the nasal passages were clear. His chest was clear to auscultation.

Hypertrophy of the cartilaginous area of the ear with sparing of the ear lobes

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Histologic examination revealed a minimal perivascular infiltrate of lymphocytes and plasma cells in the dermis with fibrosis of the subcutis (Fig. 2).

Low‐power view showing inflammation and fibrosis of the subcutis

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Blood tests showed a normal complete blood count, antinuclear antibody, and rheumatoid factor. Anti‐collagen II antibodies were elevated at 29.2 Eu/ml (normal, 0–20 Eu/ml; borderline, 20–25 Eu/ml; elevated, > 25 Eu/ml).
Seasonal effect upon excision of acquired melanocytic proliferations
Authors: Hall Glenda , Hull Cheryl , Siegel Eric R. , Smoller Bruce R. , Johnson Sandra Marchese
Published: September 2003

Background   Acquired melanocytic proliferations (AMPs) are excised throughout the year. Questions exist concerning seasonal variation in these excisions.

Objective   To investigate whether AMPs are removed more often during the spring–summer seasons than in the fall–winter and whether the percentage of benign nevi, dysplastic nevi and melanoma varies according to season.

Design   Retrospective analysis of 4794 histologically confirmed biopsies of AMPs was performed.

Results   A total of 2802 of the 4794 lesions (58%) were removed during the spring–summer season. These results confirm that seasonal variation in removal of AMPs does occur, and that removal peaks during the warmer months. In addition, the increased numbers of removals were accounted for by increases in benign pigmented lesion removals.

Conclusions   Many etiologic factors can be suggested for increased removal of AMPs during the warmer months. Reasons may include the cosmetic desires of the patient and clothing choices resulting in increased awareness of pigmented lesions. Increases in the number of skin cancer prevention programs offered by dermatologists in the spring also have a large impact on the awareness of the general population and potentially contribute to the seasonal variation.
Along these lines …
Authors: Wolf Ronni , Matz Hagit , Orion Edith
Published: September 2003

Non‐melanotic discoloration of the skin can result from the accumulation of various internal agents (ingestion, inhalation, or injection), chemicals or drugs, such as heavy metals, amiodarone, tetracycline, clofazimine and others, or exogenous (topical) substances.

Identifying the causative agent frequently requires a thorough investigation and clever detective work, including visits to the home and workplace. It is, however, rare that the patient presents at a clinic with the corpus delicti, and it is even more rare when neither the patient nor the treating physician are initially aware of it.

“What is the hardest of all? That which you hold the most simple; seeing with your own eyes what is spread out before you.”[J.W. Goethe quoted by J.P. Eckermann in Converstions with Goethe, May 18, 1824.]
Cronartium flaccidum fruitbody production on Melampyrum spp. and some important alternate hosts to pine
Authors: Kaitera By J.
Published: August 2003

Four Vincetoxicum species, Paeonia anomala and Melampyrum sylvaticum known to act as alternate hosts for Cronartium flaccidum, and a potential host Melampyrum pratense were inoculated either in vitro or in vivo with various aeciospore sources of C. flaccidum. All fresh C. flaccidum aeciospore sources variably produced uredia or telia on Vincetoxicum hirundinaria, Vincetoxicum mongolicum, Vincetoxicum fuscatum, Vincetoxicum nigrum, P. anomala, nine geographic provenances of M. sylvaticum, and two provenances of M. pratense. This is the first time that C. flaccidum telia were artificially produced on M. sylvaticum and the first time that M. pratense was shown to act as alternate host for C. flaccidum. The results suggest that M. sylvaticum is relatively susceptible to C. flaccidum, whereas M. pratense is almost resistant to this rust. The low host‐specificity of the Finnish population of C. flaccidum was demonstrated, as V. hirundinaria from Italy was susceptible to the rust.
Asymmetric formation and possible function of the primary pore canal in plutei of Temnopleurus hardwicki
Authors: Hara Yoshinobu , Kuraishi Ritsu , Uemura Isao , Katow Hideki
Published: September 2003

The development and possible function of the primary pore canal (PPC) in plutei of the sea urchin Temnopleurus hardwicki was examined by immunochemistry, electron microscopy and microsurgery. Left and right PPC that extended from coelomic sacs in plutei contained a bundle of cilia with a 9 + 2 structure that was initially detected as a group of anti‐acetylated tubulin antibody‐binding granules in the epithelium of coelomic sacs in 28 h postfertilization (PF) prism larvae. The granules extended to be a bundle of fibers toward the larval dorsal surface, concurrent with formation of the PPC on both sides, over the next 4 h. The cilia in both PPC beat actively. However, the PPC on the right side disappeared by approximately 55 h PF, establishing left–right asymmetry by 60 h PF (the four‐arm pluteus stage). The numbers of cilia in the left and right PPC in 56 h PF plutei were five and eight, respectively. Microsurgical removal of the coelomic sac from both sides or the left side only from 26 h PF prism larvae decreased body width to 64 and 91% of normal width by 50 h PF pluteus stage, respectively, whereas that of the right PPC did not. These observations suggest that PPC contribute to the maintenance of normal body width, and that there is asymmetrical activity between the left and right PPC.
Tissue sensitivity to thyroid hormone in athyroid Xenopus laevis larvae
Authors: Rot‐Nikcevic Irena , Wassersug Richard J.
Published: September 2003

Tadpoles that spontaneously arrest development and remain as larvae occur occasionally in Xenopus laevis populations. These non‐metamorphosing tadpoles continue to grow, and they develop into grossly deformed giant individuals which come as close as any anurans to being truly neotenic. Giant X. laevis tadpoles that fail to metamorphose lack thyroid glands. In this study, the hypothesis that the tissues of these tadpoles nevertheless remain thyroid hormone sensitive was tested, by exposing isolated tadpole tail tips to exogenous thyroid hormone in tissue culture. The tail tips from giant tadpoles significantly shrank in response to the thyroid hormone treatment, showing that their tissue was still capable of metamorphosis. However, the amount of shrinkage was less than that observed in tail tissue from normal tadpoles. It was hypothesized that complete induction of metamorphosis may not be possible in the giant tadpoles due to a disproportionate growth and development of tissues and organs.
Characterization of bacterial pectinolytic strains involved in the water retting process
Authors: Tamburini Elena , León Alicia Gordillo , Perito Brunella , Mastromei Giorgio
Published: August 2003

Pectinolytic microorganisms involved in the water retting process were characterized. Cultivable mesophilic anaerobic and aerobic bacteria were isolated from unretted and water‐retted material. A total of 104 anaerobic and 23 aerobic pectinolytic strains were identified. Polygalacturonase activity was measured in the supernatant of cell cultures; 24 anaerobic and nine aerobic isolates showed an enzymatic activity higher than the reference strains Clostridium felsineum and Bacillus subtilis respectively. We performed the first genotypic characterization of the retting microflora by a 16S amplified ribosomal DNA restriction analysis (ARDRA). Anaerobic isolates were divided into five different groups, and the aerobic isolates were clustered into three groups. 84.6% of the anaerobic and 82.6% of the aerobic isolates consisted of two main haplotypes. Partial 16S rRNA gene sequences were determined for 12 strains, representative of each haplotype. All anaerobic strains were assigned to the Clostridium genus, whereas the aerobic isolates were assigned to either the Bacillus or the Paenibacillus genus. Anaerobic isolates with high polygalacturonase (PG) activity belong to two clearly distinct phylogenetic clusters related to C. acetobutylicum–C. felsineum and C. saccharobutylicum species. Aerobic isolates with high PG activity belong to two clearly distinct phylogenetic clusters related to B. subtilisT and B. pumilusT.
Diversity and significance of Burkholderia species occupying diverse ecological niches
Authors: Coenye Tom , Vandamme Peter
Published: August 2003

Members of the genus Burkholderia are versatile organisms that occupy a surprisingly wide range of ecological niches. These bacteria are exploited for biocontrol, bioremediation and plant growth promotion purposes, but safety issues regarding human infections, especially in cystic fibrosis patients, have not been solved. This minireview gives an overview of the taxonomic and ecological diversity of the genus with particular emphasis on strains belonging to the Burkholderia cepacia complex and addresses the important question whether ‘good’ and ‘bad’ strains are actually the same.
Microbial community dynamics during start‐up operation of flowerpot‐using fed‐batch reactors for composting of household biowaste
Authors: Hiraishi Akira , Narihiro Takashi , Yamanaka Yosuke
Published: August 2003

Microbial community changes during start‐up operation of flowerpot‐using fed‐batch reactors for composting of household biowaste were studied by quinone profiling, rRNA‐targeted fluorescence in situ hybridization (FISH) and cultivation methods. Total and plate counts of bacteria and quinone contents in the reactors increased sharply with time during the start‐up period. These increase patterns had two phases; the first increase occurred during 3–4 weeks from the start of waste loading and the second increase was found during the subsequent 4 weeks. The microbial biomass was temporally reduced between the two succession phases. Ubiquinones predominated at the beginning of operation but decreased sharply with time, whereas partially saturated menaquinones became predominant at the fully acclimated stage. These data indicated that the major constituents of microbial populations changed from ubiquinone‐containing Proteobacteria to Actinobacteria during the period of operation. Neighbour‐joining dendrograms constructed based on the quinone profile data suggested that at least one month is required to establish a stable community structure with the Actinobacteria predominating. The characteristic population shift in the start‐up process was also demonstrated by FISH probing and 16S rDNA sequence comparisons of bacterial strains isolated.
Acidovorax‐like symbionts in the nephridia of earthworms
Authors: Schramm Andreas , Davidson Seana K. , Dodsworth Jeremy A. , Drake Harold L. , Stahl David A. , Dubilier Nicole
Published: August 2003

Dense accumulations of bacteria in the excretory organs, nephridia, were first described more than 75 years ago in members of the annelid family Lumbricidae (earthworms). These nephridial symbionts were assumed to play a role in the degradation of proteins in the excretory fluid for nitrogen recycling. In the present study, the phylogenetic affiliation of the nephridial bacteria of the earthworms Lumbricus terrestris, Aporrectodea tuberculata, Octolasion lacteum and Eisenia foetida was resolved. The 16S rRNA gene sequences of the symbionts formed a monophyletic cluster within the genus Acidovorax. Similarity between symbiont sequences from different host species was 95.5–97.6%, whereas similarity was> 99% between symbiont sequences from individuals of the same species. Densely packed bacteria were detected in the ampulla of the nephridia by fluorescence in situ hybridization (FISH) using Acidovorax‐specific oligonucleotide probes. No other bacterial cells could be found by FISH, although a few sequences other than Acidovorax had been found by PCR and cloning. These results suggest that the Acidovorax‐earthworm symbiosis is a stable, host‐specific association that has evolved from a common bacterial ancestor. Given the close phylogenetic relationship of the symbionts to proteolytic, free‐living Acidovorax species, they may indeed play a role in protein degradation during nitrogen excretion by earthworms.
Iron‐responsive genes of Phanerochaete chrysosporium isolated by differential display reverse transcription polymerase chain reaction
Authors: Assmann Eliana Maria , Ottoboni Laura Maria Mariscal , Ferraz André , Rodríguez Jaime , De Mello Maricilda Palandi
Published: August 2003

White‐rot fungus Phanerochaete chrysosporium, a ligninolytic basidiomycete, was studied to identify iron‐responsive genes. Using the differential display reverse transcription PCR technique (DDRT‐PCR), a total of 97 differentially expressed cDNA fragments were identified by comparing band intensities among fingerprints obtained from mycelia cultivated in iron‐deficient and iron‐replete media. Transcripts induced under iron‐starvation exhibited homologies to: a modular polyketide synthase, a TonB protein, a probable transmembrane protein, a putative ABC transporter permease and a HSP70‐related heat‐shock protein. Modular polyketide synthase and TonB proteins are normally expressed under iron‐starvation and are known to be involved in biosynthesis and transport of siderophores respectively. Also, a deduced protein with 96% similarity to a precursor of the well‐known P. chrysosporium lignin peroxidase was identified under iron‐deficiency. Two DDRT‐PCR products confirmed their iron‐induced expression. One was homologue to the CNOT3, which is a global regulator of RNA polymerase II transcription and has been implicated in multiple roles in the control of mRNA metabolism. The other was similar to the Schizosaccharomyces pombe putative proteasome maturation factor upm1. In conclusion, the majority of iron‐responsive P. chrysosporium transcripts isolated in the DDRT‐PCR encode proteins involved in iron acquisition, especially members of biosynthesis and transport of iron chelators.
Diversity and abundance of Crenarchaeota in terrestrial habitats studied by 16S RNA surveys and real time PCR
Authors: Ochsenreiter Torsten , Selezi Drazenka , Quaiser Achim , Bonch‐Osmolovskaya Liza , Schleper Christa
Published: August 2003

Novel phylogenetic lineages of as yet uncultivated crenarchaeota have been frequently detected in low to moderate‐temperature, marine and terrestrial environments. In order to gain a more comprehensive view on the distribution and diversity of Crenarchaeota in moderate habitats, we have studied 18 different terrestrial and freshwater samples by 16S rDNA‐based phylogenetic surveys. In seven different soil samples of diverse geographic areas in Europe (forest, grassland, ruderal) and Asia (permafrost, ruderal) as well as in two microbial mats, we have consistently found one particular lineage of crenarchaeota. The diversity of Crenarchaeota in freshwater sediments was considerably higher with respresentative 16S rDNA sequences distributed over four different groups within the moderate crenarchaeota. Systematic analysis of a 16S rDNA universal library from a sandy ecosystem containing 800 clones exclusively revealed the presence of the soil‐specific crenarchaeotal cluster. With primers specific for non‐thermophilic crenarchaeota we established a rapid method to quantify archaeal 16S rDNA in real time PCR. The relative abundance of crenarchaeotal rDNA was 0.5–3% in the bulk soil sample and only 0.16% in the rhizosphere of the sandy ecosystem. A nearby agricultural setting yielded a relative abundance of 0.17% crenarchaeotal rDNA. In total our data suggest that soil crenarchaeota represent a stable and specific component of the microbiota in terrestrial habitats.
Phylogeny of the lice (Insecta, Phthiraptera) inferred from small subunit rRNA
Authors: Barker Stephen C. , Whiting Michael , Johnson Kevin P. , Murrell Anna
Published: August 2003

There has been much argument about the phylogenetic relationships of the four suborders of lice (Insecta: Phthiraptera). Lyal's study of the morphology of lice indicated that chewing/biting lice (Mallophaga) are paraphyletic with respect to sucking lice (Anoplura). To test this hypothesis we inferred the phylogeny of 33 species of lice from small subunit (SSU) rRNA sequences (18S rRNA). Liposcelis sp. from the Liposcelididae (Psocoptera) was used for outgroup reference. Phylogenetic relationships among the four suborders of lice inferred from these sequences were the same as those inferred from morphology. The Amblycera is apparently the sister‐group to all other lice whereas the Rhynchophthirina is apparently sister to the Anoplura; these two suborders are sister to the Ischnocera, i.e. (Amblycera (Ischnocera (Anoplura, Rhynchophthirina))). Thus, the Mallophaga (Amblycera, Ischnocera, Rhynchophthirina) is apparently paraphyletic with respect to the Anoplura. Our analyses also provide evidence that: (i) each of the three suborders of lice that are well represented in our study (the Amblycera, Ischnocera, and Anoplura) are monophyletic; (ii) the Boopiidae is monophyletic; (iii) the genera Heterodoxus and Latumcephalum (Boopiidae) are more closely related to one another than either is to the genus Boopia (also Boopiidae); (iv) the Ricinidae and Laemobothridae may be sister‐taxa; (v) the Philopteridae may be paraphyletic with respect to the Trichodectidae; (vi) the genera Pediculus and Pthirus are more closely related to each other than either is to the genus Pedicinus; and (vii) in contrast to published data for mitochondrial genes, the rates of nucleotide substitution in the SSU rRNA of lice are not higher than those of other insects, nor do substitution rates in the suborders differ substantially from one another.
Proposing a solution to the Articulata–Ecdysozoa controversy
Authors: Nielsen Claus
Published: August 2003

Recent studies of animal radiation agree on monophyly of the Bilateria, but there is no consensus about the early radiation of the group. Protostomia and Deuterostomia are usually recognized, with two competing theories regarding the division of the Protostomia: one divides them into Spiralia and Cycloneuralia, the other into Lophotrochozoa and Ecdysozoa. The main discrepancy concerns the Arthropoda, which are placed with the Articulata within the Spiralia by the first group, and with the Cycloneuralia within the Ecdysozoa by the second. Here I propose that this discrepancy can be resolved by regarding the Ecdysozoa as the sister group of the Annelida within the Articulata. This implies that segmentation has been lost in phyla such as Nematoda and Priapula, but the Kinorhyncha may show a ‘reduced segmentation’ with serially arranged muscles associated with a ringed cuticle. Morphological, palaeontological and molecular implications of this theory are discussed. While many morphological and palaeontological data can be interpreted in accordance with the theory, the molecular data remain inconclusive.
COII: a useful tool for inferring phylogenetic relationships among New World monkeys (Primates, Platyrrhini)
Authors: Ascunce Marina S. , Hasson Esteban , Mudry Marta D.
Published: August 2003

In this study we evaluated the performance of the cytochromecoxidase subunit II (COII) mitochondrial gene as a tool for inferring phylogenetic relationships among platyrrhines. Twenty‐nine COII sequences were examined in seven platyrrhine genera (Alouatta,Ateles,Lagothrix,Brachyteles,Cebus,Saimiri, andAotus) employing parsimony and distance methods. Phylogenetic signal (g1) was present in all codon positions, despite the transitional saturation detected at the third position. In tree reconstructions bootstrap support values decreased abruptly above the generic level. Parsimony trees based on weighted transversions (tv : ts, 10 : 1) at the third position showed similar topologies. The utility of COII in phylogenetic studies among platyrrhines seems to be limited, due to its low rate of replacement substitutions and a relatively fast saturation of silent substitutions at third codon positions. Our data suggest that its main utility in platyrrhine systematics lies at the intrageneric level.
Testing adaptation with phylogeny: how to account for phylogenetic pattern and selective value together
Authors: Grandcolas Philippe , D’Haese Cyrille
Published: August 2003

If most agree with the dual pattern/process definition of adaptation, an apomorphy promoted by natural selection, few indeed identify an adaptation using the same methodological procedure. Most differences between procedures rest with the way the role of natural selection is analysed in a phylogenetic perspective. We argue that the selective value of a character cannot be phylogenetically reconstructed, but must be considered independently from the phylogenetic analysis in the framework of population biology. The phylogenetic pattern for the character of interest can only be used to refute or corroborate circumstantially the macro‐evolutionary predictions issuing from population studies.
A new species of Gieysztoria (Platyhelminthes, Rhabdocoela) from Argentina and a kinship analysis of South American species of the genus
Authors: Brusa Francisco , Damborenea M. Cristina , Noreña Carolina
Published: August 2003

This study describes a new species of Gieysztoria found in lentic environments in Argentina. Gieysztoria falx sp. n. appears to be closely related to G. therapaina and G. intricata, and is differentiated principally by the presence of two strongly developed pincer‐shaped spines of the cuticular stylet. A preliminary analysis of the phylogeny among the South American species of this genus is undertaken, and the monophyly verified. This study constitutes the first approach to interpreting these relationships.
Systematics, molecular genetics and historical zoogeography of the viviparous freshwater gastropodPseudopotamis(Cerithioidea, Pachychilidae): a relic on the Torres Strait Islands, Australia
Authors: Glaubrecht Matthias , Von Rintelen Thomas
Published: August 2003

Freshwater snails of the genusPseudopotamisMartens, 1894 exemplify one of the most enigmatic distributional patterns among limnic organisms in Australasia. These viviparous cerithioidean gastropods are endemic to the Torres Strait Islands, located on the Sahul shelf corridor between Australia and New Guinea. The occurrence of the two constituent species is highly restricted:P. supralirata(Smith, 1887) on Prince of Wales Island, andP. semoniMartens, 1894 on Hammond Island. Long recognized erroneously as members of the heterogeneous, polyphyletic and widespread Thiaridae, morphological and molecular data presented here reveal that the taxon has to be placed within Pachychilidae, a family otherwise absent from Australia. Comparison with other South East Asian pachychilids, in particularBrotiaH. Adams, 1866,TylomelaniaSarasin & Sarasin, 1897, andJagoraKöhler & Glaubrecht, 2003 suggests thatPseudopotamisis most closely related to taxa endemic to Sulawesi (Celebes), thus occurring disjunctly about 2000 km to the west of the Torres Strait. It is only with the Sulawesi pachychilids thatPseudopotamisshares its peculiar uterine brood pouch. Consequently, all pachychilid taxa east of the much‐debated Wallace's line (east of Bali, Borneo and the Philippines) possess this uterine incubatory structure. In this paper the historical background of the discovery, the taxonomic history, and an evaluation of the systematic position ofPseudopotamisis provided, including molecular genetic data (genes of cytochrome c oxidase subunit I and 16S ribosomal DNA). Finally, based on the phylogenetic assignment and the recent distribution, implications for a historical biogeography are discussed utilizing a recently developed model for the palaeogeography in Australasia. According to these available data, we favour early vicariant event(s) over recent dispersal.
Context of diversification of the viviparous Gyrodactylidae (Platyhelminthes, Monogenoidea)
Authors: Boeger Walter A. , Kritsky Delane C. , Pie Marcio R.
Published: August 2003

Using four criteria proposed a decade ago by Brooks & McLennan to identify a case of adaptive radiation indicates that the evolutionary history of the viviparous clade of the Gyrodactylidae is dominated by nonvicariant processes. The viviparous clade, with 446 species, has significantly more species than its sister clade (one species), and high species richness was shown to be an apomorphic trait of only the viviparous gyrodactylids within the Gyrodactylidae. Reconciliation of the phylogenetic tree of the viviparous Gyrodactylidae with that of its hosts showed a low probability for cospeciation suggesting that adaptive modes of speciation and not vicariance were predominant during the historical diversification of the clade. The proposed hypothesis suggests that the Gyrodactylidae originated on the South American continent about 60 Mya after geographical dispersal and host switching of its common ancestor to demersal freshwater catfishes by a marine ancestor. Development of hyperviviparity and the consequent loss of ‘sticky’ eggs in conjunction with other symplesiomorphic and apomorphic features allowed rapid diversification coupled with high dispersal to new host groups and geographical areas by viviparous members of the Gyrodactylidae.
Patterns of septal biomineralization in Scleractinia compared with their 28S rRNA phylogeny: a dual approach for a new taxonomic framework
Authors: Cuif Jean‐Pierre , Lecointre Guillaume , Perrin Christine , Tillier Annie , Tillier Simon
Published: August 2003

A molecular phylogeny of the Scleractinia is reconstructed from approximately 700 nucleotides of the 5′end of the 28S rDNA obtained from 40 species. A comparison of molecular phylogenic trees with biomineralization patterns of coral septa suggests that at least five clades are corroborated by both types of data. Agaricidae and Dendrophylliidae are found to be monophyletic, that is supported by microstructural data. Conversely, Faviidae and Caryophylliidae are found to be paraphyletic: Cladocora should be excluded from the faviids, whereas Eusmilia should be excluded from the caryophylliids. The conclusion is also supported by the positions, sizes and shapes of centres of calcification. The traditional Guyniidae are diphyletic, corroborating Stolarski's hypothesis ‘A’. Some results from our most parsimonious trees are not strongly statistically supported but corroborated by other molecular studies and microstructural observations. For example, in the scleractinian phylogenetic tree, there are several lines of evidence (including those from our data) to distinguish a Faviidae–Mussidae lineage and a Dendrophylliidae–Agaricidae–Poritidae–Siderastreidae lineage. From a methodological standpoint, our results suggest that co‐ordinated studies creating links between biomineralization patterns and molecular phylogeny may provide an efficient working approach for a re‐examination of scleractinian classification. This goal is important because in the evolutionary scheme proposed by Wells that presently remains the basic framework in coral studies, patterns of septal microstructures are involved. Validating from molecular phylogenies a given microstructural character state as a potential synapomorphy for a clade is the only way to include fossils in the coral classification, an approach that should allow the unity of coral classification to be maintained up to the origin of the phylum in the Triassic times.
pH‐induced alterations in stratum corneum properties
Authors: Ananthapadmanabhan K. P. , Lips A. , Vincent C. , Meyer F. , Caso S. , Johnson A. , Subramanyan K. , Vethamuthu M. , Rattinger G. , Moore D. J.
Published: August 2003

Synopsis

Skin‐cleansing compositions based on alkyl carboxylates (soaps) have a higher irritation potential than those based on syndet surfactants such as alkyl isethionates or alkyl ether sulphates. Contributing factors include inherent differences in the irritation potential of soaps and syndet surfactants, pH‐induced changes in surfactant solution chemistry, and the direct effects of pH on the physical properties of the stratum corneum (SC). Past work has not directly addressed the effect of solution pH on the SC itself and its potential role in cleanser‐induced skin irritation. In the current work, alterations to SC properties induced by buffered pH solutions and two strongly ionizable surfactants, sodium dodecyl sulphate and sodium lauryl ether sulphate, at different pH values are measured. By utilizing optical coherence tomography (OCT) and infrared (IR) spectroscopy we have directly measured physical changes in SC proteins and lipids. Our results indicate that SC swelling, which reflects alterations to SC structural proteins, is increased significantly at pH 10, compared to pH 4 and 6.5. The transition temperature (Tm) of SC lipids is found to increase at pH 10, compared to pH 4 and 6.5, suggesting a more rigid SC lipid matrix. Surfactants cause a further increase in swelling and lipid rigidity. Some aspects of what these results mean for SC physical properties as well as their implications to potential mechanisms of surfactant‐induced skin irritation are discussed.
Formulation and development of a patch containing tamarind fruit extract by using the blended chitosan–starch as a rate‐controlling matrix
Authors: Viyoch J. , Patcharaworakulchai P. , Songmek R. , Pimsan V. , Wittaya‐Areekul S.
Published: August 2003

Synopsis

A cosmetic patch containing tamarind fruit extract was formulated and developed by blending two types of natural polymers: chitosan with molecular weight of 100 000 and starch such as corn, potato or tapioca starch. The physicochemical characteristics, i.e. flexibility, colour, transparency, integrity, gloss, water sorption and bioadhesion property and the stability of the patch without tamarind content were investigated. Stability test was performed by keeping the prepared patches at 4 °C, at room temperature or at 45 °C for 2 weeks. The results showed that the formulations composed of chitosan:corn starch ratio of 4.5 : 0.5 (CC4.5 : 0.5) and chitosan:tapioca starch ratios of 4.5 : 0.5 (CT4.5 : 0.5) and 4.0 : 1.0 (CT4 : 1) provide patches with favourable physical characteristics, high water sorption, good bioadhesion ability and good stability. After the lyophilized tamarind extract in an amount corresponding to 5% of tartaric acid was incorporated into the formulations of CC4.5 : 0.5, CT4.5 : 0.5 and CT4 : 1, the ability of the patches to adhere to skin was improved. However, after keeping the test patches at room temperature or at 45 °C for 6 weeks, their colours were intensified while their flexibilities and skin adhesion properties decreased. A 12‐h in vitro permeation was investigated by studying the cumulative amount of tartaric acid permeated through the Silastic® membrane (Dow‐Coming, Midland, MI, USA). The CC4.5 : 0.5 patch tended to give the highest amount of tartaric acid released. The release pattern of all the blended polymeric matrices was exhibited in two distinct phases: the rapid phase, where the flux averaged 3.61 µg min−1 mm−2; and the slow phase, where the flux averaged 1.89 µg min−1 mm−2.
Determination of butyl methoxydibenzoylmethane, benzophenone‐3, octyl dimethyl PABA and octyl methoxycinnamate in lipsticks
Authors: Salvador A. , De La Ossa M. D. , Chisvert A.
Published: August 2003

Synopsis

The complex composition of lipstick formulations usually needs the use of organic solvents for sample dissolution. A treatment based on dissolution of cosmetic samples in ethanol‐water (70 : 30, v/v) by use of ultrasonic irradiation is proposed. A C18 stationary phase and an isocratic mobile phase of ethanol:water:acetic acid (70 : 29.5 : 0.5, v/v/v) with a flow rate of 1 mL min−1 and an injection volume of 20 µL is proposed for the high‐pressure liquid chromatography (HPLC) determination of four UV‐filters, and detection was carried out at 309 nm. The limit of the chromatographic detection was 7.0 µg mL−1 for butyl methoxydibenzoylmethane, 1.5 µg mL−1 for benzophenone‐3, 1.4 µg mL−1 for octyl dimethyl p‐aminobenzoic acid (octyl dimethyl PABA) and 2.7 µg mL−1 for octyl methoxycinnamate. The relative standard deviations of the results were between 1.8 and 2.4%. The results were comparable to those obtained by a procedure based on the use of tetrahydrofuran. The method can be considered as a green analytical procedure because it only requires the use of small quantities of ethanol.
Challenge testing: principles and practice
Authors: Russell A. D.
Published: August 2003

Synopsis

It is difficult to predict accurately the ultimate effectiveness of a preservative in any but the simplest cosmetic or pharmaceutical formulation. It is thus necessary to obtain some assurance of its likely in‐use performance in a formulated product. A challenge test is a procedure in which a product is challenged by exposure to specified types of bacteria and fungi to determine whether it is adequately preserved. Assessment of preservative efficacy is needed over the intended shelf‐life of that product. Test organisms should be representative of those likely to occur as contaminants during use and should consist of Gram‐positive and Gram‐negative bacteria, mould and yeast. In‐house factory isolates obtained as a result of contamination of earlier batches of a product may also be included. The organisms, as single or mixed inocula, are inoculated (usually as a single challenge, although some advocate multiple challenges) into samples of the product and aliquots removed at appropriate intervals for the determination of survivors. Interpretation of data is normally based on pharmacopoeial or other official protocols. Challenge testing should be undertaken at the beginning, during and at the end of the shelf‐life of the product. An alternative, the D‐value, approach is open to criticism and further studies are required that utilize rapid methods, e.g. impedance, for the detection of survivors before these can be considered to be a suitable replacement for the more traditional but very time‐consuming, viable counting procedures.
Iron sequestration on skin: a new route to improved deodorancy
Authors: Landa A. S. , Makin S. A.
Published: August 2003

Synopsis

Axillary malodour is caused by the biotransformation of non‐odorous precursors present in apocrine sweat and sebum by the axillary microflora. To counter this, underarm products typically contain high levels of bactericides. However, after an initial decrease in bacterial numbers, the surviving cells grow, producing a concomitant rise in axillary odour. A sustained deodorant effect might be achieved without recourse to bactericidal action if this bacterial growth could be inhibited for extended periods. The current study attempted to inhibit axillary bacterial growth by nutrient deprivation, primarily that of iron (Fe(III)). In vitro analyses identified iron (Fe(III)) as the trace metal whose deprivation had the most profound effect on bacterial growth. Further in vitro investigations with Fe‐chelating agents demonstrated that a number of compounds with high binding constants for Fe(III) showed optimal activity. One candidate molecule, diethylenetriaminepentaacetic acid (DTPA), was capable of effectively inhibiting bacterial growth in vitro and on the skin of the lower back. Some bacterial species could additionally utilize iron bound to the iron carrier protein transferrin present in eccrine sweat. This was minimized by the use of an agent, butylated hydroxytoluene (BHT), capable of liberating iron from transferrin via reduction of transferrin‐bound ferric ions, allowing subsequent sequestration of Fe(II). Deodorant efficacy evaluation of the combination of DTPA and BHT showed deodorancy benefits over and above that afforded by DTPA alone. This mixture of DTPA and BHT supplemented to a standard ethanolic deodorant, used on 50 people for 2 weeks, was highly effective in limiting bacterial growth in the axilla. Total aerobic bacteria in the axillae were reduced from a mean of log 5.75 (±0.73) to log 4.50 (±0.90) colony‐forming units (cfu) cm−2 (n = 27, P < 0.01) compared with a non‐fortified standard ethanolic deodorant. This was reflected in significant decreases in axillary malodour production, as determined by malodour assessments (P < 0.01). The profile of the axillary microflora was maintained, and all populations were rapidly returned to preuse levels after cessation of product use. This new deodorant technology was benchmarked against leading antimicrobial‐based deodorant systems. In three separate deodorant efficacy evaluations, the combination of DTPA and BHT was tested against Triethyl citrate, Triclosan and Farnesol in standard unfragranced ethanolic formulations. The combination of DTPA and BHT showed highly significant deodorancy benefits over and above all these antimicrobial‐based deodorant technologies. The combination of an efficient iron chelator with an agent capable of liberating iron from transferrin offers significant benefits in terms of bacterial growth inhibition on the skin and provides a new route to axillary deodorancy.
Characterization of the microflora of the human axilla
Authors: Taylor D. , Daulby A. , Grimshaw S. , James G. , Mercer J. , Vaziri S.
Published: August 2003

Synopsis

It is widely accepted that axillary malodour is attributable to the microbial biotransformation of odourless, natural secretions into volatile odorous products. Consequently, there is a need to understand the microbial ecology of the axilla in order that deodorant products, which control microbial action in this region, can be developed in the appropriate manner. A detailed characterization of the axillary microflora of a group of human volunteers has been performed. The axillary microflora is composed of four principal groups of bacteria (staphylococci, aerobic coryneforms, micrococci and propionibacteria), and the yeast genus Malassezia. Results indicated that the axillary microflora was dominated by either staphylococcal or aerobic coryneform species. Comparisons between axillary bacterial numbers and levels of axillary odour demonstrated the greatest association between odour levels and the presence of aerobic coryneforms in the under‐arm.

As the taxonomy of cutaneous aerobic coryneforms is poorly understood, a further study was conducted to characterize selected axillary aerobic coryneform isolates. Using the molecular technique of 16S rDNA sequencing, selected genomic sequences of a number of axillary aerobic coryneform isolates were obtained. Comparisons with sequence databases indicated the likely presence of a range of Corynebacterium species on axillary skin, although the majority of isolates were most similar to either Corynebacterium G‐2 CDC G5840 or C. mucifaciens DMMZ 2278. Although for a panel of individuals differences in the carriage of Corynebacterium species were noted, similar species were carried by a number of panellists. All isolates examined in this limited evaluation failed to demonstrate the capability to metabolize long‐chain fatty acids (LCFAs) to shorter chain, more volatile products. The application of this modern molecular phylogenetic technique has increased understanding of the diversity of aerobic coryneform carriage in the axilla, and on human skin. The application of this technique in other studies to assess the ethnic differences in cutaneous bacterial ecology, or the effects on the microflora of specific product use, will assist in the future development of novel deodorant systems.
Isolation, characterization and comparison of bacteria from swine faeces and manure storage pits
Authors: Cotta Michael A. , Whitehead Terence R. , Zeltwanger Rhonda L.
Published: August 2003

Storage of swine manure is associated with the microbiological production of a variety of odorous chemicals including ammonia, organic acids and alcohols, and sulphides. Although largely the product of microbiological activity, little is known about the microorganisms present in swine manure. In order to gain a better understanding of the types and activities of the microorganisms present, representative strains of microorganisms were isolated from faeces and stored manure slurry, identified, and physiologically characterized. For swine manure slurry samples, total anaerobe colony counts were greatest when a non‐selective, habitat simulating medium containing clarified swine manure slurry was used whereas the highest counts for faecal anaerobes were obtained on rumen fluid containing medium. Faecal and slurry samples were also plated onto the appropriate medium containing the antibiotics tetracycline, erythromycin and tylosin (10 µg ml−1, individually) and the proportional counts of organisms capable of growing in the presence of these antibiotics determined. Randomly selected isolates from the highest dilutions were identified by 16 s rDNA sequence analysis, and selected physiological characteristics were determined. The results of these examinations indicate that the predominant culturable microorganisms from these environments are obligately anaerobic, low mol percentage G + C Gram positive bacteria (Firmicutes) who are members of Clostridial, Eubacterial, and Lactobacillus/Streptococcus phylogenetic groups. Isolates similar to Sporomusa and Flexibacter/Cytophaga/Bacteroides (CFB or Bacteroidetes) groups were also obtained. Although similar overall, faecal and slurry samples differed in bacterial composition. Manure slurry samples were dominated by organisms similar to Clostridium coccoides and Enterococcus species whereas the distribution of species present in faeces appeared much broader. Whereas most of the pure cultures could be assigned to known phylogenetic groupings, few could be identified as known species. Examination of some growth and physiological characteristics of faecal and slurry isolates showed these to be primarily carbohydrate fermenters, although some were able to ferment lactate and amino acids. When the ability of manure and faecal isolates to ferment protein, peptides and amino acids was examined, a relatively small percentage of these were able to do so and most of these fermented carbohydrates in addition to the amino acid sources provided. The predominant amino acid fermenters were most closely related to C. coccoides and C. botulinum, but representatives of the Bacteroides, Staphylococcus, Enterococcus and other phylogenetic groups were also found. The results reported here are compared with those obtained from clone libraries prepared from the same environmental samples.
Alcanivorax which prevails in oil‐contaminated seawater exhibits broad substrate specificity for alkane degradation
Authors: Hara Akihiro , Syutsubo Kazuaki , Harayama Shigeaki
Published: August 2003

Alcanivorax is an alkane‐degrading marine bacterium which propagates and becomes predominant in crude‐oil‐containing seawater when nitrogen and phosphorus nutrients are supplemented. In order to understand why Alcanivorax overcomes other bacteria under such cultural conditions, competition experiments between Alcanivorax indigenous to seawater and the exogenous alkane‐degrading marine bacterium, Acinetobacter venetianus strain T4, were conducted. When oil‐containing seawater supplemented with nitrogen and phosphorus nutrients was inoculated with A. venetianus strain T4, this bacterium was the dominant population at the early stage of culture. However, its density began to decrease after day 6, and Alcanivorax predominated in the culture after day 20. The crude‐oil‐degrading profiles of both bacteria were therefore investigated. Alcanivorax borkumensis strain ST‐T1 isolated from the Sea of Japan exhibited higher ability to degrade branched alkanes (pristane and phytane) than A. venetianus strain T4. It seems that this higher ability of Alcanivorax to degrade branched alkanes allowed this bacterium to predominate in oil‐containing seawater. It is known that some marine zooplanktons produce pristane and Alcanivorax may play a major role in the biodegradation of pristane in seawater.
In situ distribution and activity of nitrifying bacteria in freshwater sediment
Authors: Altmann Dörte , Stief Peter , Amann Rudolf , De Beer Dirk , Schramm Andreas
Published: August 2003

Nitrification was investigated in a model freshwater sediment by the combined use of microsensors and fluorescence in situ hybridization with rRNA‐targeted oligonucleotide probes. In situ nitrification activity was restricted mainly to the upper 2 mm of the sediment and coincided with the maximum abundance of nitrifying bacteria, i.e. 1.5 × 107 cells cm−3 for ammonia‐oxidizing Beta‐proteobacteria (AOB) and 8.6 × 107 cells cm−3 for Nitrospira‐like nitrite‐oxidizing bacteria (NOB). Cell numbers of AOB decreased more rapidly with depth than numbers of NOB. For the first time, Nitrospira‐like bacteria could be quantified and correlated with in situ nitrite oxidation rates in a sediment. Estimated cell‐specific nitrite oxidation rates were 1.2–2.7 fmol NO2– cell−1 h−1.
Diversity of Bacteria and Archaea in sulphate‐reducing enrichment cultures inoculated from serial dilution of Zostera noltii rhizosphere samples
Authors: Cifuentes Ana , Antón Josefa , De Wit Rutger , Rodríguez‐Valera Francisco
Published: August 2003

We have analysed the diversity of culturable sulphate‐reducing bacteria (SRB) in Zostera noltii colonized sediments from Bassin d’Arcachon (France). Four organic substrates have been tested as well as the combination of H2 and CO2 to select for lithotrophic SRB. All energy sources were supplied in parallel cultures that were amended with yeast extract plus NH4+ and prepared without a source of combined nitrogen, the latter to select for diazotrophic SRB. The 10 different enrichment media were inoculated from serial dilution of rhizosphere samples. The highest dilution cultures yielding positive growth (i.e. 10−7) were studied by molecular techniques (16S rDNA clone libraries, RISA and ARDRA). Lactate as a single organic substrate in combination with a source of combined nitrogen resulted in selection of members of the Desulfovibrionaceae. Surprisingly, when lactate was added without a source of combined nitrogen, Desulfobacteriaceae were selected. A strong influence of the presence or absence of combined nitrogen was also observed for the substrates sucrose and fructose. Whereas the liquid culture growing on sucrose and NH4+ systematically yielded 16S rDNA clones related to an environmental unidentified green sulphur bacterium (OPS185), on plates we were able to isolate a SRB related to Desulfovibrio dechloracetivorans, which likely represents a non‐described species. Under diazotrophic conditions, sucrose selected for SRB clones related to the cluster formed by Desulfovibrio zosterae, Desulfovibrio salexigens and Desulfovibrio bastinii. The corresponding isolate obtained on plates showed only low sequence similarity with this closest neighbour (93.8%), and we suggest that it also represents a non‐described species. Surprisingly, a 16S rDNA sequence corresponding to an archaeon, i.e. a non‐extremophile Crenoarchaeota, was retrieved from several of the SRB enrichment cultures even after subsequent transfers.