A comparison between the homocyclic aromatic metabolic pathways from plant-derived compounds by bacteria and fungi

Aromatic compounds derived from lignin are of great interest for renewable biotechnical applications. They can serve in many industries e.g. as biochemical building blocks for bioplastics or biofuels, or as antioxidants, flavor agents or food preservatives. In nature, lignin is degraded by microorganisms, which results in the release of homocyclic aromatic compounds. Homocyclic aromatic compounds can also be linked to polysaccharides, tannins and even found freely in plant biomass. As these compounds are often toxic to microbes already at low concentrations, they need to be degraded or converted to less toxic forms. Prior to ring cleavage, the plant- and lignin-derived aromatic compounds are converted to seven central ring-fission intermediates, i.e. catechol, protocatechuic acid, hydroxyquinol, hydroquinone, gentisic acid, gallic acid and pyrogallol through complex aromatic metabolic pathways and used as energy source in the tricarboxylic acid cycle. Over the decades, bacterial aromatic metabolism has been described in great detail. However, the studies on fungal aromatic pathways are scattered over different pathways and species, complicating a comprehensive view of fungal aromatic metabolism. In this review, we depicted the similarities and differences of the reported aromatic metabolic pathways in fungi and bacteria. Although both microorganisms share the main conversion routes, many alternative pathways are observed in fungi. Understanding the microbial aromatic metabolic pathways could lead to metabolic engineering for strain improvement and promote valorization of lignin and related aromatic compounds.     

Osteopathy of the pectoral and pelvic limbs including pentadactyly in a young Kestrel (Falco t. tinnunculus)

Summary A five and a half weeks old female Kestrel exhibiting osteopathy of the pectoral and pelvic limbs, including symmetrical hyperdactyly, was investigated in order to clarify the pattern of the involved anatomical alterations and the possible causes of this developmental malformation. In the pectoral limb it consisted of a triplication of the alular digit, in the pelvic limb of a duplication of digit I. The live young Kestrel was observed for a period of two weeks to ascertain that it was unable to fly or procure prey on its own. After its death radiographs were taken and, apart from an eidonomic inspection including the wing claws, a detailed macroscopic dissection of the musculature of the pectoral and pelvic limbs was carried out using the in-water-method. Consecutive dissection steps were documented by a series of photographic slides. The relevant musculature, particularly that of the supernumerary digits, was recorded in proportional drawings. Subsequent to maceration of the limbs the isolated bones were reassembled according to the radiographs and also documented by means of photographs and drawings. This anatomical approach produced a reliable reconstruction of the skeletomuscular apparatus of the hyperdactylous limb parts. The eidonomic inspection revealed that at least young Kestrels may have two (alular and major digit) or even three wing claws per side. The proximal skeletal elements of both pectoral and pelvic limb were more sturdily built than in a typical Kestrel of comparable age. The proximal elements of the pelvic limb, the tarsometatarsus in particular, were shorter than in a typical Kestrel. In addition, the long axis of the tarsometatarsus was laterally bent in the transverse plane so that its proximal articular surfaces were medially inclined. Duplication of the cutaneous and osseous elements in the foot was accompanied by a duplication of some of the muscular and/or tendinous elements supplying digit I proper and the accessory digit I'. There were left-to-right asymmetries of the pedal musculature concerned. In contrast, the two accessory alular digits of each wing were almost completely devoid of musculature. Apart from atypical points of origin or insertion of the remaining distal musculture, left-to-right asymmetries and the two accessory alulae per wing, presumably, affected aerodynamic properties and resulted in flightlessness.A juvenile Kestrel of similar age and without hyperdactyly was dissected for comparison. In addition, the external appearance of the carpometacarpal region of two female Silkies, an obligatory pentadactylous breed of domestic fowl, was inspected and the skeletal parts of their pectoral and pelvic limbs compared with those of the hyperdactylous Kestrel. Our results and a literature review suggest that the symmetrical hyperdactyly in the Kestrel bears striking similarities to the hereditary hyperdactyly observed in certain breeds of domestic fowl. In addition, there is a striking resemblance between the hyperdactyly of the young Kestrel and certain forms of hyperdactyly induced by molecular genetical experiments of other authors on chicks. Comparison with these results taken from the literature suggest that the symmetrical hyperdactyly in the young Kestrel, including the alterations of the proximal skeletal elements, is caused by an unusually early expression of the Hoxd-11 gene group during embryological development. Most likely, this gene group is situated on the 2^nd chromosome in birds just as it is in mammals.

---

Osteopathie der Vorder - und Hinterextremitäten, verbunden mit einer symmetrischen Hyperdactylie bei einem jungen Turmfalken (Falco tinnunculus)

Zusammenfassung Ein fünfeinhalb Wochen alter weiblicher Turmfalke mit einer Osteopathie der Vorder- und Hinterextremitäten, verbunden mit einer symmetrischen Hyperdactylie, wurde untersucht, um das Muster der beteiligten anatomischen Veränderungen und die möglichen Ursachen dieser Mißbildung zu erkennen. An der Vorderextremität bestand sie aus einer Verdreifachung des Alula-Fingers, an der Hinterextremität aus einer Verdoppelung der Zehe I. Die Beobachtung des lebenden jungen Turmfalken während eines Zeitraumes von zwei Wochen ergab, dass er flugunfähig war und keine Beute schlagen konnte.Nach seinem Tod und einer Inspektion der Eidonomie, einschließlich der Flügelkrallen, wurden Röntgenaufnahmen angefertigt. Danach folgte eine detaillierte makroskopische Präparation der Flügel- und Beinmuskulatur unter Verwendung der In-Wasser-Methode. Die einzelnen Präparationsschritte wurden anhand von Dia-Serien dokumentiert. Die relevante Muskulatur, insbesondere die der überzähligen Digiti, wurde in proportionsgetreuen Zeichnungen festgehalten. Nach Mazeration der Extremitäten wurden die Einzelknochen, entsprechend den Röntgenbildern, wieder zusammengesetzt und ebenfalls mit Fotografien und Zeichnungen dokumentiert. Dieser anatomische Ansatz lieferte eine zuverlässige Rekonstruktion des Skelett-Muskel-Apparates der hyperdactylen Extremitätenanteile.Die eidonomische Inspektion ergab, dass zumindest junge Turmfalken zwei (Digitus alularis und majoris) oder sogar drei Flügelkrallen haben können. Die proximalen Skelettelemente der Vorder- und Hinterextremität waren deutlich robuster gebaut als bei einem typischen Turmfalken vergleichbaren Alters. Die proximalen Elemente der Hinterextremität, insbesondere der Tarsometatarsus, waren kürzer als bei einem typischen Turmfalken. Darüberhinaus war die Längsachse des Tarsometatarsus in der Transversalebene laterad gekrümmt, so dass sich seine proximalen Gelenkflächen schräg mediad richteten. Entsprechend der kutanen und knöchernen Doppelbildungen des Fußes waren auch einige der Muskeln und Sehnen doppelt vorhanden, welche die eigentliche erste Zehe und die akzessorische erste Zehe versorgten. Es traten Rechts-/Links-Asymmetrien der betreffenden Muskulatur auf. Im Gegensatz dazu waren die beiden akzessorischen Alula-Finger jedes Flügels fast vollständig ohne Muskulatur. Abgesehen von atypischen Ursprungs- und Insertionspunkten der verbleibenden distalen Muskulatur, beeinträchtigten Rechts-/Links-Asymmetrien und die beiden akzessorischen Alulae pro Flügel vermutlich die aerodynamischen Eigenschaften und führten zur Flugunfähigkeit.Ein junger Turmfalke ähnlichen Alters ohne Hyperdactylie wurde zum Vergleich präpariert. Zusätzlich wurde die äußere Erscheinung der Carpometacarpal-Region zweier Seidenhühner, einer obligatorisch pentadactylen Hühnerrasse, inspiziert und die Skelettelemente ihrer Vorder- und Hinterextremitäten mit denen des hyperdactylen Turmfalken verglichen. Unsere Ergebnisse und ein Überblick der Literatur lassen auffallende Übereinstimmungen zwischen der symmetrischen Hyperdactylie des jungen Turmfalken und der erblichen Hyperdactylie bestimmter Hühnerrassen erkennen. Darüberhinaus besteht eine auffallende übereinstimmung zwischen der Hyperdactylie des jungen Turmfalken und bestimmten Formen der Hyperdactylie, welche von anderen Autoren durch molekulargenetische Experimente an Hühnerküken induziert wurden. Ein Vergleich mit diesen Ergebnissen aus der Literatur legt nahe, dass die symmetrische Hyperdactylie des jungen Turmfalken, einschließlich der Veränderungen der proximalen Skelettelemente, durch eine ungewöhnlich frühe Expression der Hoxd-11 Gengruppe im Laufe der Embryonalentwicklung verursacht wurde. Sehr wahrscheinlich ist diese Gengruppe bei Vögeln auf dem zweiten Chromosom lokalisiert — ebenso wie bei Säugetieren.

     

Acute toxoplasmosis leads to lethal overproduction of Th1 cytokines

Virulence in Toxoplasma gondii is strongly influenced by the genotype of the parasite. Type I strains uniformly cause rapid death in mice regardless of the host genotype or the challenge dose. In contrast, the outcome of infections with type II strains is highly dependent on the challenge dose and the genotype of the host. To understand the basis of acute virulence in toxoplasmosis, we compared low and high doses of the RH strain (type I) and the ME49/PTG strain (type II) of T. gondii in outbred mice. Differences in virulence were reflected in only modestly different growth rates in vivo, and both strains disseminated widely to different tissues. The key difference in the virulent RH strain was the ability to reach high tissue burdens rapidly following a low dose challenge. Lethal infections caused by type I (RH) or type II (PTG) strain infections were accompanied by extremely elevated levels of Th1 cytokines in the serum, including IFN-gamma, TNF-alpha, IL-12, and IL-18. Extensive liver damage and lymphoid degeneration accompanied the elevated levels of cytokines produced during lethal infection. Increased time of survival following lethal infection with the RH strain was provided by neutralization of IL-18, but not TNF-alpha or IFN-gamma. Nonlethal infections with a low dose of type II PTG strain parasites were characterized by a modest induction of Th1 cytokines that led to control of infection and minimal damage to host tissues. Our findings establish that overstimulation of immune responses that are normally necessary for protection is an important feature of acute toxoplasmosis.     

Beta-carotene storage, conversion to retinoic acid, and induction of the lipocyte phenotype in hepatic stellate cells

Hepatic stellate cells (HSCs) are the major site of retinol (ROH) metabolism and storage. GRX is a permanent murine myofibroblastic cell line, derived from HSCs, which can be induced to display the fat-storing phenotype by treatment with retinoids. Little is known about hepatic or serum homeostasis of beta-carotene and retinoic acid (RA), although the direct biogenesis of RA from beta-carotene has been described in enterocytes. The aim of this study was to identify the uptake, metabolism, storage, and release of beta-carotene in HSCs. GRX cells were plated in 25 cm(2) tissue culture flasks, treated during 10 days with 3 micromol/L beta-carotene and subsequently transferred into the standard culture medium. beta-Carotene induced a full cell conversion into the fat-storing phenotype after 10 days. The total cell extracts, cell fractions, and culture medium were analyzed by reverse phase high-performance liquid chromatography for beta-carotene and retinoids. Cells accumulated 27.48 +/- 6.5 pmol/L beta-carotene/10(6) cells, but could not convert it to ROH nor produced retinyl esters (RE). beta-Carotene was directly converted to RA, which was found in total cell extracts and in the nuclear fraction (10.15 +/- 1.23 pmol/L/10(6) cells), promoting the phenotype conversion. After 24-h chase, cells contained 20.15 +/- 1.12 pmol/L beta-carotene/10(6) cells and steadily released beta-carotene into the medium (6.69 +/- 1.75 pmol/ml). We conclude that HSC are the site of the liver beta-carotene storage and release, which can be used for RA production as well as for maintenance of the homeostasis of circulating carotenoids in periods of low dietary uptake.     

Disuniting uniformity: a pied cladistic canvas of mtDNA haplogroup H in Eurasia

It has been often stated that the overall pattern of human maternal lineages in Europe is largely uniform. Yet this uniformity may also result from an insufficient depth and width of the phylogenetic analysis, in particular of the predominant western Eurasian haplogroup (Hg) H that comprises nearly a half of the European mitochondrial DNA (mtDNA) pool. Making use of the coding sequence information from 267 mtDNA Hg H sequences, we have analyzed 830 mtDNA genomes, from 11 European, Near and Middle Eastern, Central Asian, and Altaian populations. In addition to the seven previously specified subhaplogroups, we define fifteen novel subclades of Hg H present in the extant human populations of western Eurasia. The refinement of the phylogenetic resolution has allowed us to resolve a large number of homoplasies in phylogenetic trees of Hg H based on the first hypervariable segment (HVS-I) of mtDNA. As many as 50 out of 125 polymorphic positions in HVS-I were found to be mutated in more than one subcluster of Hg H. The phylogeographic analysis revealed that sub-Hgs H1*, H1b, H1f, H2a, H3, H6a, H6b, and H8 demonstrate distinct phylogeographic patterns. The monophyletic subhaplogroups of Hg H provide means for further progress in the understanding of the (pre)historic movements of women in Eurasia and for the understanding of the present-day genetic diversity of western Eurasians in general.     

Down-regulation by nuclear factor kappaB of human 25-hydroxyvitamin D3 1alpha-hydroxylase promoter

1,25-(OH)(2) vitamin D(3) is important for calcium homeostasis and cell differentiation. The key enzyme for the activation of liver-derived 25(OH) vitamin D(3) is 25-hydroxyvitamin D(3) 1alpha-hydroxylase. It is expressed mainly in the kidney but also in peripheral tissues. A 1413-bp fragment of the 1alpha-hydroxylase promoter was cloned into luciferase vectors pGL2basic and pGL3basic. Sequence analyses revealed four base exchanges and three base deletions compared with the published sequence which were identically found in five control persons. In silico promoter analyses revealed 17 putative nuclear factor (NF)kappaB sites, 10 of which were found to bind NFkappaB in EMSA experiments. Cotransfection of NFkappaB p50 and p65 subunits resulted in dramatic reduction of the promoter activity of the full-length construct as well as a series of 5'-deletion constructs. Deletion of the farmost 3'-situated NFkappaB-responsive element almost abolished NFkappaB responsiveness. Treatment of human embryonic kidney 293 cells with sulfasalazine, a NFkappaB inhibitor, resulted in enhanced 1alpha-hydroxylase mRNA production. Down-regulation of 1alpha-hydroxylase promoter through NFkappaB signaling may contribute to the pathogenesis of inflammation-associated osteopenia/osteoporosis.