Clostridium septicum gas gangrene in the orbit: a case report.

Our report presents a case of Clostridium septicum gas gangrene in an unusual, orbital localization. The predisposing factors are typical: colon tumour and lymphatic malignancy. Most probably bacteria from the intestinal flora entered the bloodstream through the compromised intestinal wall and settled in the orbit resulting in the development of an abscess containing gas. At the site of the gas gangrene, an indolent B cell lymphoma was present. After surgery and antibiotic treatment, the patient healed from the C. septicum infection; but subsequently died as a consequence of the tumour.

Neuroprotective and anticonvulsant effects of EGIS-8332, a non-competitive AMPA receptor antagonist, in a range of animal models.

BACKGROUND AND PURPOSE: Blockade of AMPA (alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid) receptors is a good treatment option for a variety of central nervous system disorders. The present study evaluated the neuroprotective and anticonvulsant effects of EGIS-8332, a non-competitive AMPA receptor antagonist, as a potential drug candidate. EXPERIMENTAL APPROACH: AMPA antagonist effects of EGIS-8332 were measured using patch-clamp techniques. Neuroprotective and anticonvulsant effects of EGIS-8332 were evaluated in various experimental models, relative to those of GYKI 53405. KEY RESULTS: EGIS-8332 inhibited AMPA currents in rat cerebellar Purkinje cells and inhibited the AMPA- and quisqualate-induced excitotoxicity in primary cultures of telencephalon neurons (IC(50)=5.1-9.0 microM), in vitro. Good anticonvulsant actions were obtained in maximal electroshock-, sound- and chemically-induced seizures (range of ED(50)=1.4-14.0 mg kg(-1) i.p.) in mice. Four days after transient global cerebral ischaemia, EGIS-8332 decreased neuronal loss in the hippocampal CA1 area in gerbils and rats. EGIS-8332 dose-dependently reduced cerebral infarct size after permanent middle cerebral artery occlusion in mice and rats (minimum effective dose=3 mg kg(-1) i.p.). Side effects of EGIS-8332 emerged much above its pharmacologically active doses. A tendency for better efficacy of GYKI 53405 than that of EGIS-8332 was observed in anticonvulsant tests that reached statistical significance in few cases, while the contrary was perceived in cerebral ischaemia tests. CONCLUSIONS AND IMPLICATIONS: EGIS-8332 seems suitable for further development for the treatment of epilepsy, ischaemia and stroke based on its efficacy in a variety of experimental disease models, and on its low side effect potential.

Prevention of congenital toxoplasmosis in Szeged, Hungary.

BACKGROUND: Toxoplasma gondii infection of the fetus can only be discovered or prevented by the appropriate serological screening and subsequent treatment of the mother and her offspring. In Hungary, there is no obligatory toxoplasma screening for pregnant women and both the reporting and follow-up of congenital toxoplasmosis cases is limited. In 1987 we started a systematic study in the Szeged region of Hungary, in which all pregnant women were screened and appropriate treatment given to all mothers and their offspring where congenital toxoplasmosis was suspected. METHODS: All pregnant women were routinely screened within the first 16 weeks of gestation for toxoplasma antibodies by complement fixation test (CFT). Seronegative cases were retested for possible seroconversion every second month. Patients with CFT titres > or = 1:256 were retested for anti-P30 immunoglobulin A (IgA), IgM and IgG antibodies by ELISA and/or SDS-PAGE-Western immunoblot in order to distinguish the acute and chronic phases of the infection. RESULTS: Up to the end of 1994, the sera of 17,735 gravidae were screened. Ten women were found to have seroconverted during pregnancy and 78 had high initial antibody levels accompanied by anti-P30 IgA antibodies at the very first screening. These two groups together were considered as definitely (10) or possibly (78) infected with Toxoplasma during pregnancy and were treated with Spiramycin. All of their offspring were also treated for one month and followed-up by systematic serological and clinical screening for 2 years. No congenital toxoplasmosis was found in any of the offspring. CONCLUSIONS: Antenatal, early diagnosis and treatment of toxoplasmosis in mothers, together with treatment and follow-up of their offspring, may considerably reduce the incidence of the disease in the offspring.

Phosphorylation of gastrin-related peptides: physiological casein kinase like enzyme in Golgi membranes from bovine adrenal chromaffin cells and GH3 cells.

Prohormones such as the gastrin precursor can be phosphorylated at Ser residues, on passage along the secretory pathway. The phosphorylation site occurs in a sequence (-Ser-Ala-Glu-) that suggests these peptides are substrates for physiological casein kinase, but the presence of this enzyme in endocrine cells is unknown. We have examined the specificity of Golgi membrane kinases from lactating rat mammary gland, bovine adrenal medulla and the GH3 cell line, for phosphorylation of progastrin fragments and analogues. The kinetics of phosphorylation of peptides with the native sequence, -Arg-Arg-Ser-Ala-Glu- were similar to those of tryptic cleavage fragments (Ser-Ala-Glu-) in both mammary and endocrine cell preparations. The product of in vitro phosphorylation was chromatographically indistinguishable from native peptide. Peptides with the sequence Ser-Ala-Ala (i.e., substitution of Glu to Ala) were not phosphorylated. We conclude that a physiological casein kinase like enzyme can act on both the gastrin precursor and its COOH-terminal cleavage product, and occurs in the Golgi complex of both mammary gland and peptide-producing endocrine cells.

[Serodiagnosis of toxoplasmosis]. / A toxoplasmosis szerodiagnosztikája.

Generally, toxoplasmosis has mild symptoms, or is asymptomatic, in patients with intact immune system. The infection, however, may have serious consequences in immunodeficient or immunosuppressed patients, as well as in the off-springs of pregnant women. If the mother has acute toxoplasmosis during the pregnancy, the passage of parasites through the placenta may result in the death of the fetus, or, in the severe damage of the fetus or neonate. All these consequences can be prevented by the early detection of the disease followed by the immediate therapy of the mother. Contrary to the most infectious diseases, however, the high specific IgM level has not proved to be a reliable marker of the acute infection in the case of toxoplasmosis. Therefore, in the case of infections discovered in the "plateau" period [i.e. with persistent IgM ("residual" IgM) and/or persistent high level of IgG antibody), the "acute" and the "chronic" phases can be distinguished more reliably by the detection with ELISA of the IgA antibody response to the so called P30 protein of Toxoplasma gondii. The anti-P30 IgA antibody response appears very early and generally disappears in 3-9 months. Thus, it is possible to discriminate the acute phase of the disease from the harmless chronic phase. Between 1987 and 1996, practically all pregnant women in Szeged and its region (altogether 21,952 women), underwent serologic toxoplasma screening. Among them, 124 pregnant women were found highly suspicious for having acute toxoplasmosis. Appropriate counselling, followed by spiramycin therapy during pregnancy and regular ultrasound examination were their antenatal management. No clinically manifested fetal or neonatal infection was observed. The screening and treatment schedule seems to be promising in the prevention of fetal and neonatal toxoplasmosis.

Apoptotic cell death induced by inhibitors of energy conservation--Bcl-2 inhibits apoptosis downstream of a fall of ATP level.

Energy charge controls intermediary metabolism and cellular regulation. Here we show that inhibition of energy conservation at the level of glucose uptake, glycolysis, citric acid cycle, and oxidative phosphorylation induces cell death, leading to fragmentation of DNA into an oligonucleosomal ladder and morphological changes typical for apoptosis. Bcl-2, the prototype of oncogenes that suppress cell death, efficiently inhibits apoptosis induced by metabolic inhibitors. Bcl-2 does not antagonize the inhibitory potential of mitochondrial inhibitors, and cannot prevent or delay the decrease of the cellular ATP level subsequent to metabolic inhibition. Thus, we propose that Bcl-2 blocks apoptosis at a point downstream of the collapse of the cellular-energy homeostasis.

Phosphorylation of human preprogastrin 93-101 by a Golgi membrane kinase from rat mammary gland.

The precursor of the acid-stimulating hormone gastrin contains a phosphorylation site which is immediately adjacent to a functionally important cleavage site, and which occurs in a sequence resembling the phosphorylation sites in casein. We have examined phosphorylation of human preprogastrin 93-101 with [gamma-32P]ATP by a Triton-solubilized Golgi membrane preparation from mammary glands of lactating rats. The activity of solubilized Golgi membranes was approx. an order of magnitude greater than that of intact vesicles suggesting a luminal orientation of the kinase. Incorporation of 32P was linear for up to 12 min at 30 degrees C, and the half-maximal rate of phosphorylation at 1 mM ATP was observed at peptide concentrations of 0.2 mM. The Km for ATP was 0.12 mM and the maximal velocity was 2.17 nmol of peptide per min per mg Golgi protein. Proteinase inhibitors (leupeptin, pepstatin, benzamidine) and p-nitrophenyl phosphate did not influence phosphorylation. The incorporation of 32P was inhibited by poly-L-lysine but not by heparin. We conclude that the phosphorylation site in progastrin is a substrate for a Golgi membrane kinase and that a similar enzyme might act on endogenous progastrin in vivo.

[Examination of the orbit using a non-ophthalmologic ultrasound equipment]. / A szemgolyó és az orbita vizsgálata nem szemészeti ultrahang készülékkel.

Ophthalmic examinations were performed for determining the suitability of seven different types of non-ophthalmic ultrasound equipments for ophthalmological purposes. According to the results, the instruments with probes of 5 MHz or higher frequency are appropriate for certain screening ophthalmological purposes in hospitals where ophthalmic ultrasound instrument is still missing. The possible indications are: approximate estimation of axial eye length for intraocular lens calculation, and visualization of gross intraocular and orbital diseases, such as retinal detachment, tumours, foreign bodies etc. In case of pathological result or in case of axial eye length different from the average value, the high resolution ophthalmological ultrasound examination is mandatory. The modern colour-coded Doppler ultrasound equipments were found suitable for the examination of the circulation of the eye and orbit.

Simplified microsurgical method of therapy for recurrent corneal erosion.

Recurrent corneal erosion can develop after superficial trauma and can prove unresponsive to traditional forms of conservative treatment. Over the past 5 years, we have treated 17 unilateral cases with our own surgical technique. The following method was applied for treatment: under an operating microscope the separated corneal epithelium was excised with microforceps and Vannas microscissors. After this, the underlying hypertrophic basement membrane was peeled off Bowman's membrane with a foreign body needle and excised at the rim with Vannas microscissors. The site of hyalin-like formation adhering on the area of Bowman's membrane was next carefully scraped with the foreign body needle. Following this procedure, a dressing lens was fitted to the eye. In 2 cases recurrences occurred, but the healing process was successful in these cases after treatment. With this simplified microsurgical procedure, the recurrent corneal erosion was curable. Through the application of dressing lenses, the danger of recurrence was eliminated, and the healing process was considerably shortened.

Unusual echographic form of retinoblastoma.

A case of an 8-year-old girl with unilateral retinoblastoma is presented. The clinical picture was dominated by inflammatory signs and retinal detachment. Ultrasonographic A/B-scan showed a funnel-shaped total retinal detachment and a thick solid mass with similar shape just behind it. The main characteristics of the mass were: irregular inside echo pattern, medium high internal reflectivity, no sign of vascularity, no after movements following a sudden ocular movement, and no shadowing of the scleral or orbital tissues behind it. The vitreous cavity in front of the retina and behind the mass was ultrasonographically clear. The described funnel-shaped echographic pattern might be typical of the diffuse infiltrating retinoblastoma.

Use of ultrasound diagnostics in lacrimal sac diseases.

Ultrasonographic examinations of the lacrimal drainage system demonstrated that the Ultrascan System IV apparatus used was not able to represent the normal-size lacrimal canaliculi, the common canaliculus, the lower part of the nasolacrimal duct and the lacrimal ostium. It can be used routinely, however, in the diagnostics of diseases of the lacrimal sac and its surrounding tissue, for surgical indication and in order to examine the postoperative condition, in most cases without other expensive diagnostic methods. The advantages over dacryocystography are that it is simple, quick, and inexpensive, it can be used post-operatively immediately, the patient is not exposed to radiation, it can be used without any contrast medium, and it is useful when the common canaliculus is occluded.

Vanadate-catalyzed, conformationally specific photocleavage of the Ca2(+)-ATPase of sarcoplasmic reticulum.

Vanadate-sensitized photocleavage of the Ca2(+)-ATPase of rabbit sarcoplasmic reticulum was observed upon illumination of sarcoplasmic reticulum vesicles or the purified Ca2(+)-ATPase by ultraviolet light in the presence of 1 mM monovanadate or decavanadate. The site of the photocleavage is influenced by the Ca2+ concentration of the medium. When the [Ca2+] is maintained below 10 nM by EGTA, the vanadate-catalyzed photocleavage yields fragments of approximately equal to 87 and approximately equal to 22 kDa, while in the presence of 2-20 mM Ca, polypeptides of 71 and 38 kDa are obtained as the principal cleavage products. These observations indicate that the site of the vanadate-catalyzed photocleavage is altered by changes in the conformation of Ca2(+)-ATPase. Selective tryptic proteolysis, at Arg-505-Ala-506, combined with covalent labeling of Lys-515 by fluorescein 5'-isothiocyanate and with the use of anti-ATPase antibodies of defined specificity, permitted the tentative allocation of the sites of photocleavage to the A fragment near the T2 cleavage site in the absence of Ca2+, and to the B fragment between Lys-515 and Asp-659 in the presence of 2-20 mM Ca2+. The loss of ATPase activity during illumination is accelerated by calcium in the presence of vanadate. The vanadate-catalyzed photocleavage in the presence of Ca2+ is consistent with the existence of an ATPase-Ca2(+)-vanadate complex (Markus et al. (1989) Biochemistry 28, 793-799).

[Ultrasound diagnosis of the lacrimal sack]. / Ultraschalldiagnostik des Tränensackes.

A- und B-scan echography is a useful method for the examination of certain parts of the lacrimal drainage system. In most cases it can substitute for dacryocystography and other expensive methods in the diagnosis of lacrimal sac diseases. Older methods of echography for examination of the lacrimal sac are direct contact, water bath, and rarely transocular contact methods. We have developed a new method, in which a rubber tube filled with fluid is placed between the transducer and the skin surface. Comparison of these methods demonstrates that this is the best method for ultrasound diagnostics of the lacrimal sac because it can be carried out as easily as the direct contact method, and the quality of the echograms is as good as in the water bath method. A special advantage is that causing an infection in the patient with the ultrasound probe can be avoided.

Cyclopiazonic acid is a specific inhibitor of the Ca2+-ATPase of sarcoplasmic reticulum.

The mycotoxin, cyclopiazonic acid (CPA), inhibits the Ca2+-stimulated ATPase (EC 3.6.1.38) and Ca2+ transport activity of sarcoplasmic reticulum (Goeger, D. E., Riley, R. T., Dorner, J. W., and Cole, R. J. (1988) Biochem. Pharmacol. 37, 978-981). We found that at low ATP concentrations (0.5-2 microM) the inhibition of ATPase activity was essentially complete at a CPA concentration of 6-8 nmol/mg protein, indicating stoichiometric reaction of CPA with the Ca2+-ATPase. Cyclopiazonic acid caused similar inhibition of the Ca2+-stimulated ATP hydrolysis in intact sarcoplasmic reticulum and in a purified preparation of Ca2+-ATPase. Cyclopiazonic acid also inhibited the Ca2+-dependent acetylphosphate, p-nitrophenylphosphate and carbamylphosphate hydrolysis by sarcoplasmic reticulum. ATP protected the enzyme in a competitive manner against inhibition by CPA, while a 10(5)-fold change in free Ca2+ concentration had only moderate effect on the extent of inhibition. CPA did not influence the crystallization of Ca2+-ATPase by vanadate or the reaction of fluorescein-5'-isothiocyanate with the Ca2+-ATPase, but it completely blocked at concentrations as low as 1-2 mol of CPA/mol of ATPase the fluorescence changes induced by Ca2+ and [ethylenebis(oxyethylenenitrilo)]tetraacetic acid (EGTA) in FITC-labeled sarcoplasmic reticulum and inhibited the cleavage of Ca2+-ATPase by trypsin at the T2 cleavage site in the presence of EGTA. These observations suggest that CPA interferes with the ATP-induced conformational changes related to Ca2+ transport. The effect of CPA on the sarcoplasmic reticulum Ca2+-ATPase appears to be fairly specific, since the kidney and brain Na+,K+-ATPase (EC 3.6.1.37), the gastric H+,K+-ATPase (EC 3.6.1.36), the mitochondrial F1-ATPase (EC 3.6.1.34), the Ca2+-ATPase of erythrocytes, and the Mg2+-activated ATPase of T-tubules and surface membranes of rat skeletal muscle were not inhibited by CPA, even at concentrations as high as 1000 nmol/mg protein.

Effect of calcium on the interactions between Ca2+-ATPase molecules in sarcoplasmic reticulum.

The interaction between Ca2+-ATPase molecules in the native sarcoplasmic reticulum membrane and in detergent solutions was analyzed by chemical crosslinking, high performance liquid chromatography (HPLC), and by the polarization of fluorescence of fluorescein 5'-isothiocyanate (FITC) covalently attached to the Ca2+-ATPase. Reaction of sarcoplasmic reticulum vesicles with glutaraldehyde causes the crosslinking of Ca2+-ATPase molecules with the formation of dimers, tetramers and higher oligomers. At moderate concentrations of glutaraldehyde solubilization of sarcoplasmic reticulum by C12 E8 or Brij 36T (approximately equal to 4 mg/mg protein) decreased the formation of higher oligomers without significant interference with the appearance of crosslinked ATPase dimers. These observations are consistent with the existence of Ca2+-ATPase dimers in detergent-solubilized sarcoplasmic reticulum. Ca2+ (2-20 mM) and glycerol (10-20%) increased the degree of crosslinking at pH 6.0 both in vesicular and in solubilized sarcoplasmic reticulum, presumably by promoting interactions between ATPase molecules; at pH 7.5 the effect of Ca2+ was less pronounced. In agreement with these observations, high performance liquid chromatography of sarcoplasmic reticulum proteins solubilized by Brij 36T or C12 E10 revealed the presence of components with the expected elution characteristics of Ca2+-ATPase oligomers. The polarization of fluorescence of FITC covalently attached to the Ca2+-ATPase is low in the native sarcoplasmic reticulum due to energy transfer, consistent with the existence of ATPase oligomers (Highsmith, S. and Cohen, J.A. (1987) Biochemistry 26, 154-161); upon solubilization of the sarcoplasmic reticulum by detergents, the polarization of fluorescence increased due to dissociation of ATPase oligomers. Based on its effects on the fluorescence of FITC-ATPase, Ca2+ promoted the interaction between ATPase molecules, both in the native membrane and in detergent solutions.

Reduction of Fe(III)ADP complex by liver microsomes.

An NADPH-driven enzymatic reduction of an Fe(III)ADP complex by rat liver microsomes has been demonstrated directly for the first time during the initial phase of lipid peroxidation by using two different analytical methods. The reduction rate increased upon increasing the ratio of ADP to ferric iron. Fe(III)ADP reducing activity of both detergent-solubilized microsomes and purified NADPH:cytochrome-P-450 (cytochrome-c) reductase decreased to about 20% compared to that of the native microsomes. Superoxide dismutase and KCN did not inhibit the reduction.

[The disease picture and treatment of Toxocara canis uveitis in childhood]. / Das Krankheitsbild und die Behandlung der Toxocara-canis-Uveitis im Kindesalter.

The disease process of uveitis due to Toxocara canis infection has a much more severe course in children than in adults. It can be cured by special drugs (Mintezol) and corticosteroids, which at the same time also reduce the inflammatory symptoms. In patients with endophthalmitis, vitrectomy may be indicated. The improvement in visual acuity depends both on the localization of the degenerations and on the duration of the disease. The authors consider sonographic examination indispensable for differential diagnosis.

Interaction of heme nonapeptide derived from cytochrome c with microsomal reductases.

The interaction of heme nonapeptide (a proteolytic product of cytochrome c) with purified NADH:cytochrome b5 (EC 1.6.2.2) and NADPH:cytochrome P-450 (EC 1.6.2.4) reductases was investigated. In the presence of heme nonapeptide, NADH or NADPH were enzymatically oxidized to NAD+ and NADP+, respectively. NAD(P)H consumption was coupled to oxygen uptake in both enzyme reactions. In the presence of carbon monoxide the spectrum of a carboxyheme complex was observed during NAD(P)H oxidation, indicating the existence of a transient ferroheme peptide. NAD(P)H oxidation could be partially inhibited by cyanide, superoxide dismutase and catalase. Superoxide and peroxide ions (generated by enzymic xanthine oxidation) only oxidized NAD(P)H in the presence of heme nonapeptide. Oxidation of NAD(P)H was more rapid with O2- than O2-2. We suggest that a ferroheme-O2 and various heme-oxy radical complexes (mainly ferroheme-O-2 complex) play a crucial role in NAD(P)H oxidation.