Genome Sequence and Characteristics of the Microbacterium foliorum Cluster EE Bacteriophage Burgy.

Burgy is a siphovirus that was isolated from compost soil near Fremont Township, Iowa, using Microbacterium foliorum NRRL B-24224. The genome has a length of 17,453 bp and contains 25 total protein-coding genes, 20 of which were assigned functions. Based on gene content, Burgy was assigned to actinobacteriophage cluster EE.

TSPO ligands stimulate ZnPPIX transport and ROS accumulation leading to the inhibition of P. falciparum growth in human blood.

After invading red blood cells (RBCs), Plasmodium falciparum (Pf) can export its own proteins to the host membrane and activate endogenous channels that are present in the membrane of RBCs. This transport pathway involves the Voltage Dependent Anion Channel (VDAC). Moreover, ligands of the VDAC partner TranSlocator PrOtein (TSPO) were demonstrated to inhibit the growth of the parasite. We studied the expression of TSPO and VDAC isoforms in late erythroid precursors, examined the presence of these proteins in membranes of non-infected and infected human RBCs, and evaluated the efficiency of TSPO ligands in inhibiting plasmodium growth, transporting the haem analogue Zn-protoporphyrin-IX (ZnPPIX) and enhancing the accumulation of reactive oxygen species (ROS). TSPO and VDAC isoforms are differentially expressed on erythroid cells in late differentiation states. TSPO2 and VDAC are present in the membranes of mature RBCs in a unique protein complex that changes the affinity of TSPO ligands after Pf infection. TSPO ligands dose-dependently inhibited parasite growth, and this inhibition was correlated to ZnPPIX uptake and ROS accumulation in the infected RBCs. Our results demonstrate that TSPO ligands can induce Pf death by increasing the uptake of porphyrins through a TSPO2-VDAC complex, which leads to an accumulation of ROS.

Effects of unilateral lesions of HVC on song patterns of male domesticated canaries.

Bird song is a model for studying neural control and lateralization of a learned behavior. Adult male canary develops large and varied song repertoires. Particular features of the male are well known to stimulate the reproductive activities of the female. We report here on the effect of lesions of either the left or right HVC, a key nucleus of the descending vocal control network of songbirds, on different song parameters of common domesticated male canaries of an European outbred strain. These canaries are useful to evaluate the question of central versus peripheral lateralization because they do not show syringeal dominance for syllable production compared to the previously studied canary strains. Right-sided lesions reduced the highest frequency and the widest frequency band. Left-sided lesion increased the lowest frequency. The size of the left-sided lesions correlated with the reduction of the repertoire of simple syllables, of the total repertoire and of the highest repetition rate, and with the increase of the lowest frequency. These results suggest a lateralized specialization of both left and right vocal pathways for particular features of the song, especially those that are known to elicit a great number of copulation solicitation displays (CSD). Lesions of both left and right pathways affected, however, sound amplitude of all syllables. Because this effect was more sever following left-sided lesions, and because the syrinx morphology of canaries has a left-right asymmetry, we suggest a peripheral mechanism for the observed lateralized specializations.

Recovery of song preferences after excitotoxic HVC lesion in female canaries.

The courtship solicitation display (CSD) of the female canary is a model to study estrogen dependent auditory preferences for male songs. The forebrain auditory-vocal nucleus, HVC, is part of the circuit that determines such preferences. To further develop this model we show that bilateral excitotoxic lesions of the medial part of HVC involving between 18-60% of the bilateral nucleus are behaviorally effective while complete unilateral lesions are not. Further, we show that animals recover their song preferences over a period of several months after the lesion. This functional recovery does not involve anatomical recovery of the HVC. Even 9 months after the lesion, the HVC size of these females was similar to that of females sacrificed 2 days after the lesion and thus was 40 +/- 8% smaller compared to normal females. Further, ipsilaterally, the lesion procedure transiently disturbed the neurochemistry, such as GAD-mRNA expression, in the part of HVC that did not undergo cell death. These results suggest that the integrity of the lateral part of at least one HVC is required to perform CSD in response to relevant auditory stimuli.

[Comparison of local and systemic inflammation after laparotomy or laparoscopy in the rat sepsis model]. / Vergleich der lokalen und systemischen Inflammation nach einer Laparotomie oder Laparoskopie im Sepsismodell der Ratte.

BACKGROUND: Laparoscopic techniques are frequently used in patients with peritonitis or intra-abdominal inflammatory diseases although increased intraperitoneal pressure may cause sepsis by promoting bacteraemia and systemic inflammatory response. METHODS: This experimental study investigates the influence of laparotomy and laparoscopy on bacteraemia, tumour necrosis factor (TNF)-alpha and endotoxin plasma levels. Standardized foecal inoculum was injected intraperitoneally and rats underwent either laparotomy (n = 20), laparoscopy (n = 20), or no further manipulation in the control group (n = 20). RESULTS: One hour after intervention, bacteraemia was significantly higher in both the laparotomy or laparoscopy groups than in the control group (P = 0.01). Foecal inoculum caused a significant increase in TNF-alpha and endotoxin plasma levels 1 h after intervention with the significantly highest levels after laparotomy (P < 0.05). In addition, the mean number of intraperitoneal abscesses were also significantly higher (P < 0.05) after laparatomy (n = 10) than after laparoscopy (n = 8) or in the control group (n = 5). CONCLUSIONS: Laparotomy and laparoscopy increased the incidence of bacteraemia and systemic inflammation compared to control group. However, inflammatory response and intraperitoneal abscess formation were significantly higher in the laparotomy group than in the laparoscopy group.

Juvenile chronic arthritis: imaging of the knees and hips before and after intraarticular steroid injection.

Intraarticular steroid therapy in juvenile chronic arthritis (JCA) is performed because of high local efficacy with few side effects. Imaging is used for initial evaluation and for monitoring of treatment response. The aim of this study was to compare imaging findings in diseased hips and knees before and after therapy. A prospective study was performed on 10 patients (15 joints) scheduled for intraarticular therapy. Pretherapeutic assessment included clinical work-up, radiographs, ultrasound (US), and magnetic resonance imaging (MRI) of affected joints. Following therapy, clinical and sonographic examinations were performed at 1 week and 1 month. MRI was repeated at 1 month. MRI and US demonstrated pannus formation and effusion, but differentiation was less distinct on US. Popliteal cysts and lymph nodes were visible in both modalities. MRI additionally revealed articular cartilage loss and subchondral cysts, not shown by US. Epiphyseal overgrowth and osteopenia were best seen radiographically. At present MRI is the best tool to assess the inflammatory changes of the joints in JCA. Initial staging of the joints may be done with plain films and MRI. US is useful to assess effusion and pannus and may be used to monitor treatment response.

Iron release from ferrisiderophores. A multi-step mechanism involving a NADH/FMN oxidoreductase and a chemical reduction by FMNH2.

Release of iron from various ferrisiderophores (ferripyoverdines, ferrioxamines B and E, ferricrocin, ferrichrome A, ferrienterobactin and its analog ferric N,N',N''-tri(1,3,5-Tris) 2,3-dihydroxybenzoylaminomethylbenzene) was obtained through an enzymic reduction of iron, involving NADH, FMN and the ferripyoverdine reductase of Pseudomonas aeruginosa PAO1. The iron released from the same complexes was also obtained through chemical reduction of iron involving FMNH2. Evidence is given that the enzymic process acts through a FMNH2 reduction; the P. aeruginosa enzyme, purified according to its ferripyoverdine-reductase activity [Hallé, F. & Meyer, J. M., Eur. J. Biochem. 209, 613-620], functions as a NADH:FMN oxidoreductase, the FMNH2 produced being able to chemically reduce the iron complexed by siderophores. The general occurrence of such a multi-step mechanism, which denies the existence of specific ferrisiderophore reductases, is discussed.

Ferrisiderophore reductases of Pseudomonas. Purification, properties and cellular location of the Pseudomonas aeruginosa ferripyoverdine reductase.

Purification of the ferripyoverdine reductase from Pseudomonas aeruginosa, strain PAO1, lead to the isolation of a soluble protein of M(r) 27,000-28,000, as determined by HPLC sieving filtration and by denaturating gel electrophoresis. In the presence of NADH as the reductant, ferripyoverdine as the iron substrate, ferrozine as an iron(II)-trapping agent and FMN, this protein displayed an iron-reductase activity which resulted in the formation of ferrozine-iron(II) complex, providing that the enzymic assay was run under strict anaerobiosis. FMN was absolutely required for the activity to occur, but the lack of a visible spectrum and the lack of fluorescence for the protein in solution suggested that ferripyoverdine reductase is not a flavin-containing protein and that covalently bound FMN is not a prerequisite for the enzymatic reaction. A search of ferripyoverdine reductase by immunological detection amongst the different cellular compartments of P. aeruginosa lead to the conclusion that the soluble enzyme, which represented more than 95% of the total cellular enzyme, is not located in the periplasm but specifically in the cytoplasm. A strongly immunoreacting material, corresponding to a protein with identical M(r) as the ferripyoverdine reductase of P. aeruginosa PAO1, was detected in all the eighteen fluorescent pseudomonad strains belonging to the P. aeruginosa, P. fluorescens, P. putida and P. chlororaphis species, as well as in P. stutzeri, a non-fluorescent species, suggesting that the enzyme acting as a ferripyoverdine reductase in P. aeruginosa PAO1 is ubiquitous among the Pseudomonas.

[Immunologic complications of visceral leishmaniasis]. / Immunologische Komplikationen bei viszeraler Leishmaniose.

This study concerns 3 cases of visceral leishmaniasis with various immunological complications. One patients showed a life-threatening activation of macrophages, characterised by haemophagocytosis in the bone marrow, pancytopenia, hypofibrinogenaemia and hypertriglycerideamia. A selective deficiency of IgG subclasses persisting for months was found in all 3 cases, the clinical relevance of which is unclear. One case manifested the already well known transient decrease of cell mediated immunity to antigens. If early detection and correct management is possible, these complications may be prevented and most cases of visceral leishmaniasis be cured.

Evaluation of methotrexate in the treatment of juvenile chronic arthritis according to the subtype.

Methotrexate therapy was evaluated in 30 children with juvenile chronic arthritis according to the type of onset. The systemic form seemed less responsive than the ANA positive form with a polyarticular course or polyarticular onset. The clinical improvement, particularly in the ANA positive polyarticular course was confirmed by a significant decrease in the values of the ESR. Side effects occurred in 12 patients and consisted of gastrointestinal upset, mouth ulcers, slight leucopenia and elevated transaminases. They led to discontinuation of the treatment in only one child. Concomitant therapy could be stopped in 50% of the patients with an ANA positive polyarticular course, but remained necessary in the two other groups. These results indicate a differential effect of MTX therapy according to the type of JCA.

Unknown syndrome: ischiadic hypoplasia, renal dysfunction, immunodeficiency, and a pattern of minor congenital anomalies.

We report a 6 year old male with a pattern of malformations and anomalies including intrauterine growth retardation, microcephaly, psychomotor retardation, a pattern of craniofacial anomalies (flat face, hypertelorism, epicanthic folds, strabismus, short nose, low set ears), hypospadias and cryptorchidism, bilateral partial cutaneous syndactyly between fingers 2 to 5 and toes 2 to 4, postaxial polydactyly of the fingers and toes, severe conductive hearing loss, hypoplasia of the ischiadic bones, complex renal dysfunction, hypogammaglobulinaemia with proneness to bacterial infections of the upper and lower respiratory tract, and recurrent pseudomembranous enterocolitis. The parents are cousins of Turkish origin.

Pharmacokinetics of pirprofen in children with juvenile chronic arthritis.

Pirprofen (100 or 200 mg; Rengasil) was administered to experimental groups of children (children with juvenile chronic arthritis, JCA) and to a control group of children (children without JCA) as a single dose or as repeated doses. The pharmacokinetics of pirprofen in these children were compared to the pharmacokinetic parameter values obtained in healthy volunteers and in elderly arthritic adults receiving 400 mg of pirprofen. The children were examined regularly and laboratory values were determined in order to detect possible side effects. The results demonstrated that the pharmacokinetics of pirprofen were similar for children and adults when taking into account the dose and the body weight. There was no drug accumulation after repeated administration of pirprofen. As already observed in rheumatic adults, pirprofen remains in synovial fluid longer than in plasma.

IgG2/IgG4 subclass deficiency in a patient with chronic mucocutaneous candidiasis and bronchiectases.

A 22-year-old man with chronic mucocutaneous candidiasis (CMC) and hypothyroidism developed severe bronchiectases following recurrent bronchopneumonia. Immunological investigations revealed IgG2/IgG4 subclass deficiency and absence of antibodies against pneumococcal and Haemophilus polysaccharides. Under regular immunoglobulin substitution every 3 weeks pulmonary symptoms improved markedly.

Cepabactin from Pseudomonas cepacia, a new type of siderophore.

In iron-deficient conditions of growth Pseudomonas cepacia ATCC 25416 excreted both pyochelin and a low-molecular-mass compound which strongly chelated iron(III), and facilitated iron translocation as demonstrated by growth and uptake experiments. The name cepabactin is proposed for this new siderophore. Comparisons of UV-visible spectra and chromatographic behaviour, together with 1H-NMR spectra, led to the conclusion that cepabactin is 1-hydroxy-5-methoxy-6-methyl-2(1H)-pyridinone, a compound which can be considered as a cyclic hydroxamate, but also as a heterocyclic analogue of catechol. This pyridinone has already been described by other workers as an antibiotic produced by Pseudomonas alcaligenes, and by a soil isolate closely related to Pseudomonas cepacia. Thus, cepabactin appears to act as a siderophore for more than one species of non-fluorescent pseudomonad.

Ferripyoverdine-reductase activity in Pseudomonas fluorescens.

Enzymatic release of iron from ferripyoverdine through a reductive mechanism was demonstrated in cell-free extracts of Pseudomonas fluorescens. Ferripyoverdine reductase activity was localized primarily in the cytoplasm and/or periplasm and appeared not to be affected by the iron status of the cells. The reaction required a strict anaerobic environment and was fully inhibited by oxygen, whereas NADH was the most effective reductant. Ferripyoverdines from other bacterial sources (P. aeruginosa ATCC 15692, P. fluorescens ATCC 13525, P. fluorescens ATCC 17400) were able to serve as iron sources as well as ferric citrate. However, the activity with ferric citrate was not strongly affected by oxygen and did not display the characteristic lag phase observed with ferripyoverdines, suggesting the occurrence of a specific ferric citrate iron reductase. FMN should play a critical role in the reductive mechanism since it was absolutely required for the activity to occur with an intensively dialyzed cell-free extract, whereas it greatly stimulated (50-fold) the NADH-mediated activity of a crude extract.