Evaluation of caspase 1 and sFas serum levels in patients with systemic sclerosis: correlation with lung dysfunction, joint and bone involvement

Recent studies point out at the role of apoptosis disturbances in the development of systemic sclerosis(SSc). The aim of our study was to examine caspase 1 and sFas serum levels in scleroderma patients and correlate the obtained results with skin involvement and internal organ changes. We studied 29 patients (14 with limited and 15 with diffuse SSc). The extension of skin involvement was measured using Total Skin Score (TSS). Internal organ involvement was assessed by specialist procedures. Serum caspase 1 and sFas levels were measured by enzyme-linked immunosorbent assay. We found correlation between sFas serum level and duration of Raynaud's phenomenon and TSS; caspase 1 serum level correlated only with TSS. Correlations between caspase 1 and lung dysfunction and sFas levels with joint and bone involvement in SSc patients were also observed. The obtained results revealed that disturbances of apoptosis might play a role in SSc pathogenesis. Caspase 1 and sFas serum levels correlate with the skin involvement severity, lung dysfunction, joint and bone changes.     

Biotransformation of waste lignin products by the soil-inhabiting yeast Trichosporon pullulans

In this study the biotransformation of lignin by-products of beechwood pulping with a soil-inhabiting yeast strain of Trichosporon pullulans was examined. The structural and molecular changes in the lignin during a cultivation process were determined by 13C NMR spectroscopy and gel permeation chromatography analysis, which confirmed the ability of the yeast strain tested to biodegrade lignin. Enzymatic analysis showed the presence of lignin peroxidase and Mn(II) peroxidase in the culture supernatant. The ligninolytic activity of both enzymes increased under carbon-depleted conditions. This observation is particularly important in the biodegradation of recalcitrant lignins in soil.     

Observation of Two Inorganic Phosphate NMR Resonances in the Perfused Hypothermic Rat Heart

The effect of hypothermia on isolated perfused rat hearts was studied with³¹P NMR. Hearts were continuously perfused with phosphate-free Krebs–Henseleit buffer while the perfusate temperature was adjusted. Perfusate pH was kept at 7.40 ± 0.02 throughout the experiments. Using the chemical shift difference between PCr and P_ithe intracellular pH was estimated. At 36, 20, and 10°C a cytosolic alkalinization at a pH of 7.05 ± 0.04, 7.21 ± 0.05, and 7.40 ± 0.03 was observed, respectively. At 10°C two P_iresonances were observed with a separation of 0.25 ppm. This resonance corresponded to a P_iresonance of a cellular compartment with a local pH of 7.78 ± 0.06, likely mitochondrial. This additional resonance disappeared upon warming of the hearts back to 36°C.     

The effect of chemical treatment of stainless steel wire surfaces on Zymomonas mobilis cell attachment and product synthesis

The attachment, growth and product synthesis of non-flocculating Zymomonas mobilis cell, fixed in stainless steel wire spheres (WS), were investigated. The carrier surface was activated by treatment with titanium (IV) chloride (TiCl₄) and γ-aminopropyltriethoxysilane (AS) in an attempt to raise the efficiency in the immobilization of the cells. System productivity for ethanol and levan production, using cells immobilized on a modified stainless steel in the batch fermentation of a sucrose medium, rose as a result of increased biomass compared to the productivity of cells fixed on untreated (control) metal surfaces. Stabilized ethanol synthesis was demonstrated in the course of four cycles (each cycle 48 h) of repeated fermentations with a stainless steel carrier treated with AS, and three cycles when TiCl₄ was used. Levan synthesis decreased after three cycles with cells immobilized on a silanized surface. System productivity for ethanol and levan production after the fourth cycle in experiments with TiCl₄-activated, silanized and unmodified carriers were Q_eth = 1.01, 1.06 and 0.27 g/l × h; Q_lev = 0.32, 0.29 and 0.12 g/l × h, respectively. However, the specific productivity of biomass for product synthesis was higher in fermentation systems with untreated stainless steel surfaces, probably due to some loss of physiological activity of cells attached to a modified carrier. Investigations of throughly washed activated stainless steel wire surfaces, by scanning electron microscopy after immobilization, showed significant attachment of cells to the carriers. A polymer layer covered the wire surface treated with TiCl₄ after fermentations. This may be explained as the binding of extracellular polysaccharide, such as the fructose-polymer levan and yeast extract components, to the modified support via chelation. After four fermentations, craters and holes in the polymer layer were evident, probably as a result of CO₂ formation. A small number of cells appeared on this layer. In view of the good ethanol formation during all fermentation cycles, it is probably that active Z. mobilis cells remained under the polymer layer. Wire treatment with AS resulted in the formation of long filamentous cells during fermentation and some disturbance of cellular fission. This may be partly explained by strong electrostatic interactions between the positively charged carrier surface and the predominately negatively charged surface of Z mobilis cells. However, this did not significantly affect other cellular functions. The surface of the wire treated with AG was practically without a polymer layer.     

Allergenic pollen and pollinosis in Warsaw

Summary Our study of fifty two hay fever patients included twenty six solely allergic to grass pollen and twenty six exhibiting allergy to various pollen species, such as hazel, birch, oak, poplar, andArtemisia. Their total and specific IgE response was evalutated by the immunoenzymatic method, while clinical reactivity was assessed by recording nasal and bronchial symptom scores between mid-March and mid-July. Simultaneously pollen counts were made. Polysensitized patients showed significantly higher levels of both total and specific IgE, which testifies to the enhanced quantitative and qualitative IgE. Multisensitized patients reacted earlier than patients sensitized to grass pollen only, which confirms that non-grass plants flowering only in the spring cause the priming effect on the nasal and bronchial mucosa. The early symptoms may be attributable to tree pollen sensitivity or may refletct higher grass pollen IgE levels in the polysensitized group. Characteristically, nasal symptoms preceded bronchial symptoms of several weeks.On comparing nasal washing from the polysensitized patients to washing from patients with grass pollen, we found much cytological material with the predominance of eosinophils.     

Protein a of quinolinate synthetase is the site of oxygen poisoning of pyridine nucleotide coenzyme synthesis in Escherichia coli

De novo biosynthesis of pyridine nucleotide coenzymes in Escherichia coli is initiated by an enzyme complex (quinolinate synthetase) containing protein B which converts l-aspartate into iminoaspartate protein A, which then generates quinolinate on the pathway to the coenzymes. This complex has been shown to be poisoned by hyperbaric oxygen. ^7,8 We performed assays made dependent on both proteins B and A versus only protein A, using cell-free extracts of hyperbaric-oxygen poisoned and aerobically grown cells. The specific activities were produced by a similar amounts of 68% and 60%, respectively, when measured in assays made dependent on enzymes B and A versus only protein A that was derived from oxygen-poisoned extract. Thus, protein A is the oxygen-sensitive component.     

Porin threading drives receptor disengagement and establishes active colicin transport through Escherichia coli OmpF

Bacteria deploy weapons to kill their neighbours during competition for resources and to aid survival within microbiomes. Colicins were the first such antibacterial system identified, yet how these bacteriocins cross the outer membrane (OM) of Escherichia coli is unknown. Here, by solving the structures of translocation intermediates via cryo‐EM and by imaging toxin import, we uncover the mechanism by which the Tol‐dependent nuclease colicin E9 (ColE9) crosses the bacterial OM. We show that threading of ColE9’s disordered N‐terminal domain through two pores of the trimeric porin OmpF causes the colicin to disengage from its primary receptor, BtuB, and reorganises the translocon either side of the membrane. Subsequent import of ColE9 through the lumen of a single OmpF subunit is driven by the proton‐motive force, which is delivered by the TolQ‐TolR‐TolA‐TolB assembly. Our study answers longstanding questions, such as why OmpF is a better translocator than OmpC, and reconciles the mechanisms by which both Tol‐ and Ton‐dependent bacteriocins cross the bacterial outer membrane.     

Multiple forms of Pregnancy-Associated Glycoproteins released in vitro by porcine chorion or placentomal and interplacentomal explants of wild and domestic ruminants

Characterization of the Pregnancy-Associated Glycoproteins (PAG) is important for studies of reproduction of various eutherian domestic, wild and endangered mammals. Distinct chorionic PAG genes are expressed in embryo-origin cells: pre-placental trophoblast (TR) and in placental trophectoderm (TRD) of various entherians. This study demonstrates in vitro production of the PAG proteins during long-term cultures of various chorionic explants: porcine TR or TRD, cotyledonary (CT) of European bison (Eb), and CT or intercotyledonary (intCT)-TRD of the cattle. Chorionic proteins isolated from media were analyzed by homologous or heterologous Western immunoblotting with anti-PAG sera, raised against cellular bovine or secretory porcine antigens. Used anti-PAG sera identified diverse molecular forms of released PAG proteins: 43-69 kDa for EbPAG proteins, 40-85 kDa for bovine PAG (bPAG), and 43-73 kDa for porcine PAG (pPAG). Immunoblotting revealed also that both CT and intCT-TRD explants secreted equivalent amounts of bPAG proteins. This useful system of in vitro protein production can provide native chorionic PAG proteins with placental unique carbohydrate chains. The PAG proteins are required as standard markers for diagnostic tests of pregnancy in domestic and wild mammals, in which seasonal reproductive processes are relatively difficult to control.