Changes in extracellular acid-base homeostasis in cerebral ischemia

The purpose of this study was to examine the changes in extracellular CO ₃ ²⁻ and lactate concentration produced by ischemia, especially in relation to the occurrence of anoxic depolarization, and how some of these changes are altered by the inhibition of organic acid transport systems with probenecid. These data demonstrate that (i) the transmembrane mechanisms contributing to intracellular acid-base regulation (Na⁺/H⁺ and HCO ₃ ⁻ /Cl⁻ exchanges, and lactate/H⁺ cotransport) are markedly activated during ischemia; (ii) the efficacy of these mechanisms is abolished as the cellular membrane permeability to ions, including H⁺ and pH-changing anions, suddenly increases with anoxic depolarization; and (iii) efflux of intracellular lactate during ischemia, and its reuptake with reperfusion, mainly occur via a transporter. These findings imply that residual cellular acid-base homeostasis persists as long as cell depolarization does not occur, and strengthen the concept that anoxic depolarization is a critical event for cell survival during ischemia. Special issue dedicated to Dr. Herman Bachelard.     

Production and Characterization of Monoclonal Antibodies against Vegetative Cells of Bacillus cereus

Two monoclonal antibodies (MAbs) against Bacillus cereus were produced. The MAbs (8D3 and 9B7) were selected by enzyme-linked immunosorbent assay for their reactivity with B. cereus vegetative cells. They reacted with B. cereus vegetative cells while failing to recognize B. cereus spores. Immunoblotting revealed that MAb 8D3 recognized a 22-kDa antigen, while MAb 9B7 recognized two antigens with molecular masses of approximately 58 and 62 kDa. The use of MAbs 8D3 and 9B7 in combination to develop an immunological method for the detection of B. cereus vegetative cells in foods was investigated.     

Sex-dependent antioxidant enzyme activities and lipid peroxidation in ageing mouse brain

We investigated whether oxidant status and antioxidant enzyme activities during ageing of mouse brain are regulated in sex-dependent manner. In the homogenate from the brain of 1, 4, 10 and 18 months old male and female CBA mice, lipid peroxidation (LPO), total superoxide dismutase (tSOD), catalase (CAT) and glutathione peroxidase (Gpx) were determined. LPO was age- and sex-related, favoring males over females throughout the lifespan with the peak in both sexes at 10 months of age. Throughout ageing, no difference in tSOD activity between male and female brains was observed, except in immature 1 month old mice. Gender-related difference in Gpx activity was observed, with higher level in females comparing to males, reaching statistical significance in senescent (18 months old) animals. CAT activity was drastically changed with ageing in both the male and female brain. We found different age associated trends in CAT activity in males and females: decreased with age in males and increased with age in females. Taken together, the present findings indicate that brains of female mice have lower oxidant and higher antioxidant capacity mostly related to CAT and to a lesser extent to Gpx activity.     

Genetic structure and expansion of golden jackals (Canis aureus) in the north-western distribution range (Croatia and eastern Italian Alps)

The golden jackal, widely distributed in Europe, Asia and Africa, is one of the less studied carnivores in the world and the genetic structure of the European populations is unknown. In the last century jackals strongly declined mainly due to human persecution, but recently they expanded again in eastern Europe. With the aim to determine the genetic structure and the origin of expanding jackals, we analyzed population samples obtained from Bulgaria, Serbia, Croatia (Dalmatia and Slavonia) and individuals sampled in north-eastern Italy. Samples were typed at the hypervariable part of the mitochondrial DNA control-region (mtDNA CR1) and at 15 canine autosomal microsatellite loci (STR), and analyzed using multivariate, Bayesian and landscape genetic methods. The mtDNA CR1 was monomorphic, showing a single haplotype shared among all the populations. The STR loci were variable, with 2–14 alleles and intermediate values of heterozygosity (Ho = 0.47; He = 0.51). Genetic diversity was significantly partitioned (θ_ST = 0.07; P < 0.001) and the populations were partially distinct, perhaps in consequence of recent fragmentations. Jackals from Dalmatia were the most genetically differentiated. Assignment testing and gene flow analyses suggested that jackals colonizing Italy have admixed origins from Dalmatian and Slavonian populations. They are not first generation migrants, suggesting that dispersal towards north-eastern Italy is a stepping-stone process. Golden jackal and wolf colonization patterns might be different, with prevalent short-distance dispersal in jackals versus prevalent long distance dispersal in wolves. The admixed origin of jackals in the Alps ensures abundant genetic variability, which may enhance adaptive fitness and expectancy of population growth. The intersections between Dinaric–Balkan and Eastern Alps are areas of population expansion and admixture, highlighting their conservation, ecological and evolutionary values.     

Production and characterization of monoclonal antibodies against vegetative cells of Bacillus cereus

Two monoclonal antibodies (MAbs) against Bacillus cereus were produced. The MAbs (8D3 and 9B7) were selected by enzyme-linked immunosorbent assay for their reactivity with B. cereus vegetative cells. They reacted with B. cereus vegetative cells while failing to recognize B. cereus spores. Immunoblotting revealed that MAb 8D3 recognized a 22-kDa antigen, while MAb 9B7 recognized two antigens with molecular masses of approximately 58 and 62 kDa. The use of MAbs 8D3 and 9B7 in combination to develop an immunological method for the detection of B. cereus vegetative cells in foods was investigated.     

Production of extracellular lipases by Penicillium cyclopium purification and characterization of a partial acylglycerol lipase

Penicillium cyclopium, grown in stationary culture, produces a type I lipase specific for triacylglycerols while, in shaken culture, it produces a type II lipase only active on partial acylglycerols. Lipase II has been purified by ammonium sulfate precipitation and chromatographies on Sephadex G-75 and DEAE-Sephadex. The enzyme exists in several glycosylated forms of 40-43 kDa, which can be converted to a single protein of 37 kDa by enzymatic deglycosylation. Activity of lipase II is maximal at pH 7.0 and 40 degrees C. The enzyme is stable from pH 4.5 to 7.0. Activity is rapidly lost at temperatures above 50 degrees C. The enzyme specifically hydrolyzes monoacylglycerols and diacylglycerols, especially of medium chain fatty acids. The sequence of the 20 first amino acid residues is similar to the N-terminal region of P. camembertii lipase and partially similar to lipases from Humicola lanuginosa and Aspergillus oryzae, but is different from Penicillium cyclopium lipase I. However, it can be observed that residues of valine and serine at positions 2 and 5 in Penicillium cyclopium lipase II are conserved in Penicillium expansum lipase, of which 16 out of the 20 first amino acid residues are similar to Penicillium cyclopium lipase I.     

Treatment of pseudophakic cystoid macular oedema

Cystoid oedema of macula is the most frequent problem following non-complicated cataract surgery, which leads to weakening of visual acutity. We have studied the treatment outcomes of the topical therapy with corticosteroids and nonsteroid antirheumatics during prospective randomized study in comparison with orally applied acethazolamide. However, 8 weeks later in treatment was observed a positive effect in both groups, on the eyes of patients, treated with acetazolamide, resorption of the oedema was faster, more intensive and ultimate visual acutity was statistically significantly better. Results demonstrated a benefit of combined therapy with antiinflamative drugs and orally prescribed acetazolamide in pseudophakic macular oedema.     

Diet spectrum of great cormorants (<Emphasis Type="Italic">Phalacrocorax carbo sinensis</Emphasis> L.) at the Donji Miholjac carp fishponds in eastern Croatia

This paper reports the results of a diet analysis from the digestive tracts of 203 great cormorants shot at the Donji Miholjac fishponds in eastern Croatia, in the period 2000–2002. Eight fish species were determined. The dominant species was common carp (Cyprinus carpio) with a relative frequency of 73.4%, followed by grass carp (Ctenopharingodon idella) (11.6%), bighead carp (Aristichthys nobilis) (7.2%), pikeperch (Sander lucioperca) (2.0%), wels catfish (Silurus glanis) (1.7%), pike (Esox lucius) (1.5%), topmouth gudgeon (Pseudorasbora parva) (1.5%) and Prussian carp (Carassius auratus gibelio) (1.1%). The diet spectrum of the great cormorants from the Donji Miholjac fishponds was in accordance with the structure of the fish population in the fishponds. The average stomach weight per bird was 244 g. When birds shot with an empty stomach were excluded, the average stomach weight increased to 286 g. The length of consumed fish ranged from 40–335 mm, with 47% of the fish belonging to the length category 100–149 mm. The study revealed no significant relationship between the weight of the consumed fish and the body weight of male and female cormorants.