Local dynamic stability in turning and straight-line gait

Successful community and household ambulation require the ability to navigate corners and maneuver around obstacles, posing unique challenges compared to straight-line walking. The challenges associated with turning may contribute to an increased incidence of falling and the occurrence of fall-related injuries. A measure of stability applied to turning gait may be able to quantify a system's response to naturally occurring disturbances associated with turning and identify subjects at greater risk of falling. An index of stability has been used previously to assess the rate of kinematic separation (local dynamic stability) during straight-line gait. The purpose of this study was to determine if local dynamic stability during constant speed turning is reduced compared to straight-line treadmill walking. Maximum finite-time Lyapunov exponents (λ) were used to estimate the local stability of able-bodied subjects’ (n=19) sagittal plane hip, knee, and ankle trajectories for turning compared to straight-line walking at two different walking speeds. Turning λ was greater than straight λ for the hip, right knee, and ankle (p<0.05). Turning λ for the left knee angle was similar to straight λ. There were no differences in λ between left and right limbs for the hip and ankle and also no differences between the inside and outside limbs during turning for all joints. These findings indicate able-bodied subjects’ hip, right knee, and ankle kinematics are less locally stable while turning than walking in a straight line and may be used as a comparative tool for determining the efficacy of therapeutic interventions for mobility-impaired populations.     

Spin-crossover iron(II) complexes [Fe(Medpq)(py)2(NCS)2] and [Fe(Medpq)(py)2(NCSe)2]: syntheses, characterization and magnetic properties

Two new spin-crossover complexes, [Fe(Medpq)(py)₂(NCS)₂] · py · 0.5H₂O (1) and [Fe(Medpq)(py)₂(NCSe)₂] · py (2) (Medpq = 2-methyldipyrido[3,2-f:2′,3′-h]-quinoxaline, py = pyridine), have been synthesized. The crystal structures were determined at both room temperature (298 K) and low temperature (110 K). Complexes 1 and 2 crystallize in the orthorhombic space group Pbca and monoclinic space group P2₁/n, respectively. In both complexes, the distorted [FeN₆] octahedron is formed by six nitrogen atoms from Medpq, the trans pyridine molecules and the cis NCX⁻ groups. The thermal spin transition is accompanied by the shortening of the mean Fe–N distances by 0.194 Å for 2. The mononuclear [Fe(Medpq)(py)₂(NCS)₂] and [Fe(Medpq)(py)₂(NCSe)₂] neutral species interact each other via π-stacking, resulting in a one-dimensional extended structure for both 1 and 2. There exist C–HX (X = S, Se) hydrogen bonds for both complexes. Variable-temperature magnetic susceptibility measurements and Mössbauer spectroscopy reveal the occurrence of a gradual spin transition. The transitions are centered at T_1/2 = 120 K for 1 and T_1/2 = 180 K for 2, respectively.     

Structural determinations, magnetic and EPR studies of complexes involving the Cr(OH)2Cr unit

Five heterometallic compounds with formulae [Ba(H₂O)₄Cr₂(μ-OH)₂(nta)₂] · 3H₂O (I), [M(bpy)₂(H₂O)₂] [Cr₂(OH)₂(nta)₂] · 7H₂O, where M²⁺ = Zn, (II); Ni, (III); Co, (IV) and [Mn(H₂O)₃(bpy)Cr₂(OH)₂(nta)₂] · (bpy) · 5H₂O (V); bpy = 2,2′-bipyridine, (nta = nitrilotriacetate ion) have been prepared by reaction of I with the corresponding M^II-sulfates in the presence of 2,2′-bipyridine. Substances I–V have been characterized by magnetic susceptibility measurements, EPR and X-ray determinations. I represents a 2D coordination polymer formed by coordination of centrosymmetrical dimeric chromium(III) units and Barium cations. The 10-coordinate Ba polyhedron is completed by four water molecules. Compounds II–IV are isostructural and consist of non-centrosymmetric dimeric anions [Cr₂(μ-OH)₂(nta)₂]²⁻, complex cations [M^II(bpy)₂(H₂O)₂]²⁺ and solvate water molecules. The octahedral coordination of chromium atoms implies four donor atoms of the nta³⁻ ligands and two bridging OH groups. Multiple hydrogen bonds of coordinated and solvate water molecules link anions and cations in a 3D network. A similar [Cr₂(μ-OH)₂(nta)₂]²⁻ unit is found in V. The bridging function is performed by a carboxylate oxygen atom of the nta ligand that leads to the formation of a trinuclear complex [Mn(bpy)(H₂O)₂Cr₂(μ-OH)₂(nta)₂]. Experimental and calculated frequency and temperature dependences of EPR spectra of these compounds are presented. The fine structure appearing on the EPR spectra of compound V is analyzed in detail at different temperatures. It is established that the main part of the EPR signals is due to the transitions in the spin states of a spin multiplet with S = 2. Analyses of experimental and calculated spectra confirm the absence of interaction between metal ions (M^II) and Cr-dimers in complexes III and IV and the presence of weak Mn–Cr interactions in V. The temperature dependence of magnetic susceptibilities for I–V was fitted on the basis of the expression derived from isotropic Hamiltonian including a bi-quadratic exchange term.     

Static magnetic fields: animal studies

Various experimental studies carried out over the last 30–40 years have examined the effects of the chronic or acute exposure of laboratory animals to static magnetic fields. Many of the earlier studies have been adequately reviewed elsewhere; few adverse effects were identified. This review focuses on studies carried out more recently, mostly those using vertebrates, particularly mammals. Four main areas of investigation have been covered, viz., nervous system and behavioural studies, cardiovascular system responses, reproduction and development, and genotoxicity and cancer. Work on the role of the natural geomagnetic field in animal orientation and migration has been omitted.

Generally, the acute responses found during exposure to static fields above about 4 T are consistent with those found in volunteer studies, namely the induction of flow potentials around the heart and the development of aversive/avoidance behaviour resulting from body movement in such fields. No consistently demonstrable effects of exposure to fields of 1 T and above have been seen on other behavioural or cardiovascular endpoints. In addition, no adverse effects of such fields on reproduction and development or on the growth and development of tumours have been firmly established. Overall, however, far too few animal studies have been carried out to reach any firm conclusions.     

Haplotypes of nine single nucleotide polymorphisms on chromosome 19q13.2-3 associated with susceptibility of lung cancer in a Chinese population

To evaluate the joint effect of nine single nucleotide polymorphisms for three DNA repair genes in the region of chromosome 19q13.2-3 on susceptibility of lung cancer in a Chinese population, we conducted a hospital-based case–control study consisting of 247 lung cancer cases and 253 cancer-free controls matched on age, gender and ethnicity. Associations between the haplotypes and susceptibility of lung cancer were tested. The global test of haplotype association revealed a statistically significant difference in the haplotype distribution between cases and controls (global test: χ² = 60.45, d.f. = 15, P = 2.11E−07). The two haplotypes were underrepresented among cases (Hap5 defined by ERCC1118^A–ERCC2156^C–ERCC2312^G–ERCC2751^A–XRCC1194^T–XRCC1206^A–XRCC1280^G–XRCC1399^G–XRCC1632^G and Hap12 defined by ERCC1118^G–ERCC2156^C–ERCC2312^G–ERCC2751^A–XRCC1194^C–XRCC1206^A–XRCC1280^G–XRCC1399^A–XRCC1632^G). Three of the haplotypes were overrepresented among cases (Hap3 defined by ERCC1118^A–ERCC2156^C–ERCC2312^G–ERCC2751^A–XRCC1194^C–XRCC1206^A–XRCC1280^G–XRCC1399^G–XRCC1632^G, Hap4 defined by ERCC1118^A–ERCC2156^C–ERCC2312^G–ERCC2751^A–XRCC1194^C–XRCC1206^G–XRCC1280^G–XRCC1399^G–XRCC1632^A, and Hap10 defined by ERCC1118^G–ERCC2156^A–ERCC2312^G–ERCC2751^A–XRCC1194^T–XRCC1206^A–XRCC1280^G–XRCC1399^G–XRCC1632^G). Haplotypes 3 and 10 (cases = 5.7%, controls = 1.0%, OR = 6.56, 95%CI = 1.83–23.54, P = 0.001; cases = 13.3%, controls = 5.6%, OR = 2.73, 95%CI = 1.51–4.94, P = 0.0006) were the most strongly associated with increased lung cancer risk. There was considerable linkage disequilibrium exists between SNPs both within genes and between genes in the region. The two blocks for solid spine of LD and six htSNPs were found. The haplotype analysis suggested that the biologically effective polymorphisms co-segregate with some of the haplotypes. This result supports the hypothesis that the sub-region is important for lung cancer susceptibility. Haplotype studies using larger study groups will be required to obtain conclusive results.     

Preparation and characterization of immobilized lipase on magnetic hydrophobic microspheres

A novel magnetic poly(vinyl acetate (VAc)–divinyl benzene (DVB)) material (8–34 μm) was synthesized by copolymerization of vinyl acetate and divinyl benzene using oleic acid-stabilized magnetic colloids as magnetic cores. The magnetic colloids and the copolymer microspheres were characterized with transmission and scanning electron microscopes, respectively. Magnetization of the microspheres could be described by the Langevin function. All the observations indicated that the microspheres were superparamagnetic. Magnetic sedimentation of the microspheres was achieved within 3 min, over 300 times faster than the gravitational sedimentation. Candida cylindracea lipase (CCL) was immobilized to the porous carrier at up to 6750 IU/g carrier, remarkably higher than the previous studies. The pH and temperature dependencies of the immobilized CCL were investigated and the optimum temperature and pH for the immobilized CCL were determined. Activity amelioration of the immobilized CCL for the hydrolysis of olive oil was observed, indicating an interfacial activation of the enzyme after immobilization. Moreover, the immobilized CCL showed enhanced thermal stability and good durability in the repeated use after recovered by magnetic separations.     

Critical exponents for sol–gel transition in aqueous alginate solutions induced by cupric cations

The sol–gel transition in aqueous alginate solutions of four alginate samples having different molecular weights (M_W) and M/G ratios induced by cupric cations was monitored by rheology measurements. The gel point f_gel and the relaxation critical exponent n were determined using the Winter’s criterion over the alginate concentration C_Alg of 1–4 wt%. The scaling for the zero shear viscosity η₀ before the gel point and the equilibrium modulus G_e after the gel point was established against the relative distance ε from the gel point at the concentration of C_Alg = 1 wt%, giving the critical exponents k and z. The results indicated that f_gel was almost independent of the alginate concentration and became higher for the sample with lower molecular weight. The critical exponent n decreased with the increase in C_Alg for these four Cu-alginate samples and the fractal dimension d_f estimated from n suggested a denser structure in the critical gel with high G content. The critical exponent n evaluated from k and z agreed well with n determined from the Winter’s criterion.     

The site of action of magnetic stimulation of human motor cortex in a patient with motor neuron disease

Cortical and spinal root (C₈-T₁) magnetic stimulation evoked single motor unit potentials in the abductor pollicis brevis and abductor digiti minimi muscles in a patient with chronic motor neuron disease. This patient was unique in that there were few surviving motoneurons in these muscles making it possible to record single motor units. Central motor conduction time was within normal limits. Cortical mapping of motor evoked potentials (MEPs) was carried out using a twin magnetic coil stimulating over the motor cortex at intervals of 1 cm along the coronal axis and 1–2 cm along the sagittal axis. As the site of cortical stimulation was moved from the centre, the latency of the MEPs increased by 0.7–0.8 ms suggesting one synaptic delay. This study provides further data that magnetic stimulation of the human cortex indirectly activates pyramidal cells via interneurons.     

Suppression of the endoplasmic reticulum calcium pump during zebrafish gastrulation affects left-right asymmetry of the heart and brain

Vertebrate embryos generate striking Ca²⁺ patterns, which are unique regulators of dynamic developmental events. In the present study, we used zebrafish embryos as a model system to examine the developmental roles of Ca²⁺ during gastrulation. We found that gastrula stage embryos maintain a distinct pattern of cytosolic Ca²⁺ along the dorsal–ventral axis, with higher Ca²⁺ concentrations in the ventral margin and lower Ca²⁺ concentrations in the dorsal margin and dorsal forerunner cells. Suppression of the endoplasmic reticulum Ca²⁺ pump with 0.5 μM thapsigargin elevates cytosolic Ca²⁺ in all embryonic regions and induces a randomization of laterality in the heart and brain. Affected hearts, visualized in living embryos by a subtractive imaging technique, displayed either a reversal or loss of left–right asymmetry. Brain defects include a left–right reversal of pitx2 expression in the dorsal diencephalon and a left–right reversal of the prominent habenular nucleus in the brain. Embryos are sensitive to inhibition of the endoplasmic reticulum Ca²⁺ pump during early and mid gastrulation and lose their sensitivity during late gastrulation and early segmentation. Suppression of the endoplasmic reticulum Ca²⁺ pump during gastrulation inhibits expression of no tail (ntl) and left–right dynein related (lrdr) in the dorsal forerunner cells and affects development of Kupffer’s vesicle, a ciliated organ that generates a counter-clockwise flow of fluid. Previous studies have shown that Ca²⁺ plays a role in Kupffer’s vesicle function, influencing ciliary motility and translating the vesicle’s counter-clockwise flow into asymmetric patterns of gene expression. The present results suggest that Ca²⁺ plays an additional role in the formation of Kupffer’s vesicle.     

A new floating sensor array to detect electric near fields of beating heart preparations

A new flexible sensor for in vitro experiments was developed to measure the surface potential, Φ, and its gradient, E (electric near field), at given sites of the heart. During depolarisation, E describes a vector loop from which direction and magnitude of local conduction velocity θ can be computed. Four recording silver electrodes (14 μm × 14 μm) separated by 50 μm, conducting leads, and solderable pads were patterned on a 50 μm thick polyimide film. The conductive structures, except the electrodes, were isolated with polyimide, and electrodes were chlorided. Spacer pillars mounted on the tip fulfil two functions: they keep the electrodes 70 μm from the tissue allowing non-contact recording of Φ and prevent lateral slipping. The low mass (9.1 mg) and flexibility (6.33 N/m) of the sensor let it easily follow the movement of the beating heart without notable displacement. We examined the electrodes on criteria like rms-noise of Φ, signal-to-noise ratio of Φ and E, maximum peak-slope recording dΦ/dt, and deviation of local activation time (LAT) from a common signal and obtained values of 24–28 μV, 46 and 41 dB, 497–561 V/s and no differences, respectively. With appropriate data acquisition (sampling rate 100 kHz, 24-bit), we were able to record Φ and to monitor E and θ on-line from beat-to-beat even at heart rates of 600 beats/min. Moreover, this technique can discriminate between uncoupled cardiac activations (as occur in fibrotic tissue) separated by less than 1 mm and 1 ms.     

The effect of two piglet teeth resection procedures on the welfare of sows in farrowing crates. Part 2

Recent EU legislation discourages the practice of resecting piglets’ needle teeth. However, the effect of leaving piglets’ teeth intact on the welfare of sows in farrowing crates is poorly understood. Therefore, the aim of the study was to compare the effects of clipping and grinding piglets’ needle teeth, compared to leaving them intact, on the welfare of sows in farrowing crates.

Six days pre-partum 60 multiparous sows were assigned to one of three treatments. Litters had their teeth clipped (C), ground (G) or left intact (I) at birth. Sows’ teats were inspected for lesions pre-partum (day −3) and on days 1, 4, 11, 18 and 27 post-partum. Instantaneous scan samples (5 min intervals) of sow behaviour were carried out during three 2 h periods on days 1, 4, 8, 14, 21 and 26. On days 1, 4 and 11 all piglets were removed from the crate for 60 min. On re-introduction of the piglets, sow maternal behaviour was recorded continuously for 20 min.

The number of sows with teat lesions tended to differ between treatments on days 11 (P = 0.06) and 18 (P = 0.10). There was an interactive effect between treatment and day on sow dog-sitting behaviour throughout lactation (P < 0.001) and a tendency for an interactive effect on posture-changing behaviour (P = 0.08). On day 21, I sows were dog-sitting in more observations than C sows (P < 0.05) and on day 26 in more observations than C and G sows (P < 0.001). There was an interaction between treatment and day in the latency of sows to suckle their piglets following 60 min separation (P < 0.05). On day 4, I sows had a shorter latency to suckle than C and G sows (P < 0.05). There was an effect of treatment on the number of sows that terminated bouts of post-suckling udder massage (P < 0.05). On day 4, more I than C sows terminated post-suckling udder massage (P = 0.01). Finally, there was an effect of treatment on the time spent lying in the ventral posture in the observations of maternal behaviour (P < 0.05). C sows spent less time lying in the ventral posture than G and I sows (P < 0.05).

There were indications that leaving the teeth intact and to a lesser extent grinding caused injury and disturbance to sows. In farrowing crates, leaving piglets’ teeth intact cannot be recommended.     

Synthesis, spectral studies and in vitro assessment for antiamoebic activity of new cyclooctadiene ruthenium(II) complexes with 5-nitrothiophene-2-carboxaldehyde thiosemicarbazones

We report here the synthesis, characterization and in vitro antiamoebic activity of 5-nitrothiophene-2-carboxaldehyde thiosemicarbazones (TSC), 1–5, and their bidentate complexes [Ru(η⁴-C₈H₁₂)(TSC)Cl₂] 1a–5a. The biological studies of these compounds were investigated against HK-9 strain of Entamoeba histolytica and the concentration causing 50% cell growth inhibition (IC₅₀) was calculated in the micromolar range. The ligands exhibited antiamoebic activity in the range (2.05–5.29 μM). Screening results indicated that the potencies of the compounds increased by the incorporation of ruthenium(II) in the thiosemicarbazones. The complexes 1a–5a showed antiamoebic activity with an IC₅₀ of 0.61–1.43 μM and were better inhibitors of growth of E. histolytica, based on IC₅₀ values. The most promising among them is Ru(II) complex 2a having 1,2,3,4-tetrahydroquinoline as N⁴ substitution.     

Chronic exposure to sublethal doses of radiation mimetic Zeocin selects for clones deficient in homologous recombination

Research indicates that exposure to hypoxia is associated with oxidative stress. In this investigation, healthy subjects were exposed to hypoxia by inhalation of 10% oxygen for 2 h (corresponding to 5500 m above sea level). The levels of strand breaks and oxidatively damaged purine bases, measured by the comet assay, and the expression of genes involved in DNA repair of oxidatively damaged DNA were investigated in mononuclear blood cells (MNBC) at baseline, after 2 h of hypoxia, 2 h of reoxygenation, and 1 day and 8 days after the exposure. The level of strand breaks and oxidized purine bases in MNBC increased following both the 2 h of hypoxia and the 2 h reoxygenation period, whereas this effect was not observed in unexposed subjects. The expressions of oxoguanine DNA glycosylase 1 (OGG1), nucleoside diphosphate linked moiety X-type motif 1 (NUDT1), nei endonuclease VIII-like 1 (NEIL1), and mutY homolog (MUTYH) were unaltered throughout the experiment in both groups of subjects, indicating that DNA repair genes are not up-regulated by the hypoxia and reoxygenation treatment. Taken together, this report shows that inhalation of 10% oxygen for 2 h is associated with increased number of oxidized DNA lesions in MNBC, but acute hypoxia may not inflict upon the regulation of genes involved in repair of oxidized DNA.     

Endothelial nitric oxide synthase gene haplotypes and circulating nitric oxide levels significantly associate with risk of essential hypertension

Nitric oxide (NO), a potent vasodilator, plays a pivotal role in blood pressure regulation. Endothelial NO synthase gene (NOS3) polymorphisms influence NO levels. Here, we investigated the role of the – 922A/G, – 786T/C, 4b/4a, and 894G/T polymorphisms of the NOS3 and NO_x levels in 800 consecutive unrelated subjects comprising 455 patients of essential hypertension and 345 controls. The polymorphisms were investigated independently and as haplotypes. Plasma NO_x levels (nitrate and nitrite) were estimated by the Griess method. Genotype frequencies for the –786T/C, 4b/4a, and 894G/T polymorphisms differed significantly (P < 0.001) between patients and controls and were associated with an increased risk of hypertension (OR = 2.0, OR = 3.8, OR = 1.6, respectively). The 4-locus haplotypes ATaG (H1), ATaT (H2), and GCaG (H3) were significantly associated with essential hypertension and served as susceptible haplotypes (P ≤ 0.0001). On the other hand, haplotypes ATbG (H4) and GTbG (H5) were negatively associated with hypertension and served as protective haplotypes (P < 0.0001). NO_x levels were significantly lower in patients than controls (P < 0.0001). The individual polymorphisms showed marginal association with NO_x level; however, the susceptible haplotype H2 associated significantly with lower NO_x levels in patients (P < 0.001) and conversely the haplotype H4 with higher NO_x levels in controls (P < 0.001). In conclusion, the 4b/4a and likely – 786T/C polymorphisms were identified as the determinants modifying the risk of hypertension. This study identifies the NOS3 variants and haplotypes as genetic risk factors and as useful markers of increased susceptibility to the risk of essential hypertension.     

DNA damage and repair in gastric cancer--a correlation with the hOGG1 and RAD51 genes polymorphisms

Aristolochic acid (AA) is a potent nephrotoxin and carcinogen and is the causative factor for Chinese herb nephropathy. AA has been associated with the development of urothelial cancer in humans, and kidney and forestomach tumors in rodents. To investigate the molecular mechanisms responsible for the tumorigenicity of AA, we determined the DNA adduct formation and mutagenicity of AA in the liver (nontarget tissue) and kidney (target tissue) of Big Blue rats. Groups of six male rats were gavaged with 0, 0.1, 1.0 and 10.0 mg AA/kg body weight five times/week for 3 months. The rats were sacrificed 1 day after the final treatment, and the livers and kidneys were isolated. DNA adduct formation was analyzed by ³²P-postlabeling and mutant frequency (MF) was determined using the λ Select-cII Mutation Detection System. Three major adducts (7-[deoxyadenosin-N⁶-yl]-aristolactam I, 7-[deoxyadenosin-N⁶-yl]-aristolactam II and 7-[deoxyguanosin-N²-yl]-aristolactam I) were identified. There were strong linear dose-responses for AA-induced DNA adducts in treated rats, ranging from 25 to 1967 adducts/10⁸ nucleotides in liver and 95–4598 adducts/10⁸ nucleotides in kidney. A similar trend of dose-responses for mutation induction also was found, the MFs ranging from 37 to 666 × 10⁻⁶ in liver compared with the MFs of 78–1319 × 10⁻⁶ that we previously reported for the kidneys of AA-treated rats. Overall, kidneys had at least two-fold higher levels of DNA adducts and MF than livers. Sequence analysis of the cII mutants revealed that there was a statistically significant difference between the mutation spectra in both kidney and liver of AA-treated and control rats, but there was no significant difference between the mutation spectra in AA-treated livers and kidneys. A:T → T:A transversion was the predominant mutation in AA-treated rats; whereas G:C → A:T transition was the main type of mutation in control rats. These results indicate that the AA treatment that eventually results in kidney tumors in rats also results in significant increases in DNA adduct formation and cII MF in kidney. Although the same treatment does not produce tumors in rat liver, it does induce DNA adducts and mutations in this tissue, albeit at lower levels than in kidney.     

Metal–salt co-tolerance and metal removal by indigenous cyanobacterial strains

Chromium and salt tolerance in five indigenous cyanobacterial strains isolated from contaminated sites was investigated along with their metal bioaccumulative potential. All the five species showed significantly better growth when the medium was spiked with salt or chromium. As compared to single metal or salt treatment, the binary metal–salt (MS) treatments had more favorable effect on cyanobacterial growth as indicated by significantly higher concentration of the primary photosynthetic pigment chlorophyll at M₂₀S₂₀₀₀ (9.9–25.3 μg/mL) as compared to that at M₀S₀ (4.0–12.3 μg/mL). Similarly biomass was much higher at M₂₀S₁₀₀₀ and M₂₀S₂₀₀₀ (41.8–86.2 mg/10 mL) as compared to that at control, M₀S₀ (21.5–36.3 mg/10 mL). Accessory pigments like carotenoids and phycobilinproteins too tended to increase significantly in response to both metal and salts in the two species of Lyngbya (L. putealis and L. ceylanica var. constricta) and Gloeocapsa. These species also showed greater potential of chromium bioaccumulation, which increased further as both salt and metal concentration increased. In the two species of Nostoc however, bioaccumulative potential improve at higher metal concentration, but not affected significantly by salt concentration.     

Thermo-alkali-stable catalases from newly isolated Bacillus sp. for the treatment and recycling of textile bleaching effluents

Three thermoalkaliphilic bacteria, which were grown at pH 9.3–10 and 60–65 °C were isolated out of a textile wastewater drain. The unknown micro-organisms were identified as thermoalkaliphilic Bacillus sp. Growth conditions were studied and catalase activities and stabilities compared. Catalases from Bacillus SF showed high stabilities at 60 °C and pH 9 (t_1/2=38 h) and thus this strain was chosen for further investigations, such as electron microscopy, immobilization of catalase and hydrogen peroxide degradation studies. Degradation of hydrogen peroxide with an immobilized catalase from Bacillus SF enabled the reuse of the water for the dyeing process. In contrast, application of the free enzyme for treatment of bleaching effluents, caused interaction between the denaturated protein and the dye, resulting in reduced dye uptake, and a higher color difference of 1.3 ΔE* of dyed fabrics compared to 0.9 ΔE* when using the immobilized enzyme.     

Synthesis and in vitro anti-hepatitis B and C virus activities of ring-expanded ('fat') nucleobase analogues containing the imidazo[4,5-e][1,3]diazepine-4,8-dione ring system

As part of our structure–activity relationship studies, we report here the synthesis and in vitro anti-HBV and anti-HCV activities of a number of ring-expanded (‘fat’) nucleobases containing the imidazo[4,5-e][1,3]diazepine-4,8-dione ring system. One of the compounds, ZP-88, exhibited a good activity/toxicity profile against HBV by inhibition of the synthesis of extracellular virion release (EC₅₀ = 1.7 μM, CC₅₀ = 286 μM, SI = 168) and intracellular HBV replication intermediates (EC₅₀ = 8.4 μM, CC₅₀ = 286 μM, SI = 34) in cultured human hepatoblastoma 2.2.15 cells. By contrast, most of the compounds tested against HCV had only marginal activity/toxicity profile, although that was still better than that of the reference compound ribavirin.     

Pullulans produced by strains of Cryphonectria parasitica—II. Nuclear magnetic resonance evidence

The structure of pullullan-like polysaccharides produced as exocellular material by different strains of Cryphonectria parasitica, the fungus responsible for chestnut tree cankers, was investigated with nuclear magnetic resonance (NMR) techniques. ¹³C, mono- and bidimensional ¹H, and ¹H–¹³C heteronuclear correlated NMR spectra (HSQC and HMBC) were recorded. Advanced analysis of the NMR spectra allowed the main resonance of the atoms in the maltotriose and in the maltotetraose repeat units of pullulan-like polysaccharides from C. parasitica to be recognised with confidence. In all cases investigated, the presence of large amounts of -(1→6) maltotetraose subunits was evidenced, in addition to the -(1→6) maltotriose subunits, corresponding to the repeating unit of pullulan produced by Aureobasidium pullulans and other fungi. The results were in agreement with other data from this laboratory, obtained with independent techniques. The belief that in ‘pullulans’ the maximum amount of -(1→6) maltotetraose subunits is about 7% can thus be considered as definitely outdated.     

Hsp70 response in pigs is affected by their Halothane genotypes after heat stress

 Heat shock protein ([Hsp70]) and ionized calcium ([Ca²⁺]) concentrations were examined in pigs’ blood at different times after heat stress (HS). Pigs were from different halothane genotypes (HAL) (NN, n=7; Nn, n=5; nn, n=6). [Hsp70] increased significantly from 1–2 h after HS and [Ca²⁺] decreased significantly from HS-end in NN and Nn. No significant differences in [Hsp70] and [Ca²⁺] were detected among NN and Nn. [Hsp70] and [Ca²⁺] in nn did not vary significantly from baseline values. [Hsp70] was significantly higher in NN and Nn than in nn. Briefly, HAL affected [Hsp70] after HS in pigs probably via a disordered [Ca²⁺] regulation.