Structure of coenzyme F420H2 oxidase (FprA), a di-iron flavoprotein from methanogenic Archaea catalyzing the reduction of O2 to H2O

The di-iron flavoprotein F(420)H(2) oxidase found in methanogenic Archaea catalyzes the four-electron reduction of O(2) to 2H(2)O with 2 mol of reduced coenzyme F(420)(7,8-dimethyl-8-hydroxy-5-deazariboflavin). We report here on crystal structures of the homotetrameric F(420)H(2) oxidase from Methanothermobacter marburgensis at resolutions of 2.25 A, 2.25 A and 1.7 A, respectively, from which an active reduced state, an inactive oxidized state and an active oxidized state could be extracted. As found in structurally related A-type flavoproteins, the active site is formed at the dimer interface, where the di-iron center of one monomer is juxtaposed to FMN of the other. In the active reduced state [Fe(II)Fe(II)FMNH(2)], the two irons are surrounded by four histidines, one aspartate, one glutamate and one bridging aspartate. The so-called switch loop is in a closed conformation, thus preventing F(420) binding. In the inactive oxidized state [Fe(III)FMN], the iron nearest to FMN has moved to two remote binding sites, and the switch loop is changed to an open conformation. In the active oxidized state [Fe(III)Fe(III)FMN], both irons are positioned as in the reduced state but the switch loop is found in the open conformation as in the inactive oxidized state. It is proposed that the redox-dependent conformational change of the switch loop ensures alternate complete four-electron O(2) reduction and redox center re-reduction. On the basis of the known Si-Si stereospecific hydride transfer, F(420)H(2) was modeled into the solvent-accessible pocket in front of FMN. The inactive oxidized state might provide the molecular basis for enzyme inactivation by long-term O(2) exposure observed in some members of the FprA family.     

Human Semaphorin 3 Variants Link Melanocortin Circuit Development and Energy Balance

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Strain-specific Influence of Microcystis Aeruginosa on Food Ingestion and Assimilation of some Cladocerans and Copepods

Ingestion rates where estimated for daphnids, Cyclops spp. and Bosmina (Eubosmina) coregoni thersites fed hepatotoxic and non-toxic M. aeruginosa either separate or mixed with the readily available food alga Ankistrodesmus falcatus. The ingestion rates of hepatotoxic strains of M. aeruginosa are very low compared with those of A. falcatus or non-toxic M. aeruginosa HUB 5-3 fed to Daphnia magna or D. longispina. However, a close relationship between ingestion rate of different M. aeruginosa strains and their toxicity could not be observed. Addition of the toxic strain M. aeruginosa HUB 5-2-4 reduces the ingestion rates of A. falcatus progressively due to increased food rejection by D. magna. Additionally, the assimilation efficiency of M. aeruginosa HUB 5-2-4 is two times lower compared with A. falcatus and M. aeruginosa HUB 5-3 leading to strong starvation.     

Daphnia and Toxic Blooms of Microcystis aeruginosa in Bautzen Reservoir (GDR)

As a part of a whole-lake, long-term experiment in biomanipulation in. the hypertrophic Bautzen reservoir (G.D.R.), during three years (1984–1986) the dynamics of mouse-related LD 50 of Microcystis aeruginosa was compared with the biomass development of this blue-green and the grazing pressure exerted by Daphnia galeata. Since the three summer averages of the biomass of D. galeata revealed strong differences due to decreasing predation activity of fish from 1984 to 1986, the effects of different grazing pressure on Microcystis toxicity could be investigated under field conditions. Microcystis was nontoxic at the beginning of the growing season and developed high toxicity during its first strong biomass increase in summer in all three years. But this decrease of the LD 50 together with the first biomass increase of the season is found in quite different periods in different years (1984: August, 1985: July, 1986: June). It is obvious that the higher the mean effective filtration rate of D. galeata during summer is found the faster the toxicity of Microcystis is formed. If these observations are combined with findings of other authors, the conclusion can be drawn that the development of toxic Microcystis blooms seems to be promoted by a combination of five conditions: (1) Presence of a mixture of toxic and nontoxic Microcystis strains at the beginning of the growing season even if the portion of toxic strains is very low, (2) physical and chemical growth conditions which favour Microcystis over other phytoplankton, (3) high grazing pressure by zooplankton on edible food particles over a rather long period, (4) patchy distribution of the different Microcystis strains if nonselective filtrators such as Daphnia dominate the zooplankton, and (5) absence of defense mechanisms of Microcystis against grazing which are not coupled with toxicity (e.g. large colony size). These conclusions contribute to a better understanding of the possibilities and limits of in-lake eutrophication control by biomanipulation and emphasize the need to combine top-down and bottom-up control mechanisms in eutrophic and hypertrophic waters.     

Inhibition of major histocompatibility complex class II-dependent antigen presentation by neutralization of gamma interferon leads to breakdown of resistance against measles virus-induced encephalitis

BALB/c mice are resistant to measles virus (MV)-induced encephalitis due to their strong MV-specific CD4(+) T-cell response. Resistance is broken by neutralization of gamma interferon with monoclonal antibodies, indicating an important role for this pleiotropic cytokine. Here, we demonstrate that mouse gamma interferon has no direct antiviral effect in vitro and in vivo. The breakdown of resistance is due neither to a switch in the T-helper response nor to an impaired migration of CD4(+) T cells. Neutralization of gamma interferon interferes with the major histocompatibility complex class II-dependent antigen presentation and subsequent proliferation of CD4(+) T cells in vitro and in vivo. In consequence, the reduction in numbers of CD4(+) T cells below a protective threshold leads to susceptibility to MV-induced encephalitis.     

Targeting the immune system: novel therapeutic approaches in squamous cell carcinoma of the head and neck

Despite advances in surgery, radiotherapy, and chemotherapy, the overall survival rates for patients with squamous cell carcinoma of the head and neck (SCCHN) have not changed over the last decades. Clearly, novel therapeutic strategies are needed for this cancer, which is highly immunosuppressive. Therefore, biologic therapies able to induce and/or up-regulate antitumor immune responses could represent a complementary approach to conventional treatments. Because patients with SCCHN are frequently immunocompromised due to the elimination or dysfunction of critical effector cells of the immune system, it might be necessary to restore these immune functions to allow for the generation of more effective antitumor host responses. Simultaneously, to prevent tumor escape, it might be necessary to alter attributes of the malignant cells. The present review summarizes recent advances in the field of immunotherapy of SCCHN, including techniques of nonspecific immune stimulation, the use of monoclonal antibodies, advances in adoptive immunotherapy and genetic engineering, as well as anticancer vaccines. These biologic therapies, alone or in combination with conventional treatment, are likely to develop into useful future treatment options for patients with SCCHN.     

Distribution of Chitinases in the Entomopathogen <Emphasis Type="Italic">Metarhizium anisopliae</Emphasis> and Effect of <Emphasis Type="Italic">N</Emphasis>-Acetylglucosamine in Protein Secretion

For a long time, fungi have been characterized by their ability to secrete enzymes, mostly hydrolytic in function, and thus are defined as extracellular degraders. Chitin and chitinolytic enzymes are gaining importance for their biotechnological applications. Particularly, chitinases are used in agriculture to control plant pathogens. Metarhizium anisopliae produces an extracellular chitinase when grown on a medium containing chitin, indicating that synthesis is subject to induction by the substrate. Various sugar combinations were investigated for induction and repression of chitinase. N-acetylglucosamine (GlcNAc) shows a special dual regulation on chitinase production. M. anisopliae has at least two distinct, cell-bound, chitinolytic enzymes when cultured with GlcNAc as one of the carbon sources, and we suggest that this carbohydrate has an important role in protein secretion.     

M protein, a classical bacterial virulence determinant, forms complexes with fibrinogen that induce vascular leakage

Increased vascular permeability is a key feature of inflammatory conditions. In severe infections, leakage of plasma from the vasculature induces a life-threatening hypotension. Streptococcus pyogenes, a major human bacterial pathogen, causes a toxic shock syndrome (STSS) characterized by excessive plasma leakage and multi-organ failure. Here we find that M protein, released from the streptococcal surface, forms complexes with fibrinogen, which by binding to beta2 integrins of neutrophils, activate these cells. As a result, neutrophils release heparin binding protein, an inflammatory mediator inducing vascular leakage. In mice, injection of M protein or subcutaneous infection with S. pyogenes causes severe pulmonary damage characterized by leakage of plasma and blood cells. These lesions were prevented by treatment with a beta2 integrin antagonist. In addition, M protein/fibrinogen complexes were identified in tissue biopsies from a patient with necrotizing fasciitis and STSS, further underlining the pathogenic significance of such complexes in severe streptococcal infections.     

Growth and enzyme production by three Penicillium species on monosaccharides

The growth and preference for utilisation of various sugar by the Penicillium species Penicillium pinophilum IBT 4186, Penicillium persicinum IBT 13226 and Penicillium brasilianum IBT 20888 was studied in batch cultivations using various monosaccharides as carbon source, either alone or in mixtures. P. pinophilum IBT 4186 and P. persicinum IBT 13226 had a micro(max) around 0.08-0.09 h(-1) using either glucose or xylose as carbon source. The micro(max) of P. brasilianum IBT 20888 was 0.16 and 0.14 h(-1) on glucose and xylose, respectively. Glucose was found to exert repression on the catabolism of mannose, galactose, xylose and arabinose. The three species were able to utilise all the tested monosaccharides, but arabinose was only slowly metabolised. Glucose was also found to repress the production of endoglucanases, endoxylanases and beta-xylosidases. After glucose depletion, the fungi started producing beta-glucosidase and endoglucanases. Xylose did not repress the enzyme production and it induced the production of endoxylanases and beta-xylosidases.