Impact of subunit and oligomeric structure on the thermal and conformational stability of chlorite dismutases

Chlorite dismutases (Cld) are unique heme b containing oxidoreductases that convert chlorite to chloride and dioxygen. Recent phylogenetic and structural analyses demonstrated that these metalloproteins significantly differ in oligomeric and subunit structure. Here we have analyzed two representatives of two phylogenetically separated lineages, namely pentameric Cld from Candidatus "Nitrospira defluvii" and dimeric Cld from Nitrobacter winogradskyi having a similar enzymatic activity at room temperature. By application of a broad set of techniques including differential scanning calorimetry, electronic circular dichroism, UV-vis and fluorescence spectroscopy the temperature-mediated and chemical unfolding of both recombinant proteins were analyzed. Significant differences in thermal and conformational stability are reported. The pentameric enzyme is very stable between pH 3 and 10 (T(m)=92°C at pH 7.0) and active at high temperatures thus being an interesting candidate for bioremediation of chlorite. By contrast the dimeric protein starts to unfold already at 53°C. The observed unfolding pathways are discussed with respect to the known subunit structure and subunit interaction.     

N9-benzyl-substituted 1,3-dimethyl- and 1,3-dipropyl-pyrimido[2,1-f]purinediones: synthesis and structure-activity relationships at adenosine A1 and A2A receptors

Synthesis and physicochemical properties of N-benzyl pyrimido[2,1-f]purinediones are described. These derivatives were synthesized by the cyclization of 7-chloropropylo-8-bromo-1,3-dimethyl- or 1,3-dipropyl xanthine derivatives with corresponding (un)substituted benzylamines. Dipropyl derivatives were obtained under microwave irradiation conditions either. The obtained compounds (1-20) were evaluated for their affinity to adenosine A1 and A2A receptors, selected compounds were additionally investigated for affinity to the A3 receptor subtype. The results of the radioligand binding assays to A1 and A2A adenosine receptors showed that most of the 1,3-dimethyl-9-benzylpyrimidopurinediones exhibited selective affinity to A2A receptors at micromolar or submicromolar concentrations (for example, derivative 9 with o-methoxy substituent displayed a Ki value of 0.699 microM at rat A2A receptor with more than 36-fold selectivity). Contrary to previously described arylpyrimido[2,1-f]purinediones dipropyl derivatives (compounds 15-20) showed affinity to both kinds of receptors increased, however A1 affinity increased to a larger extent, with the result that A2A selectivity was abolished. The best adenosine A1 receptor ligand was m-chlorobenzyl derivative 18 (Ki=0.089 microM and 5-fold A1 selectivity). Structure-activity relationships were discussed with the analysis of lipophilic and spatial properties of the investigated compounds. Pharmacophore model of adenosine A1 receptor antagonist was adopted for this purpose.     

Study of the reactions between platinum(II) complexes and L-methionine in the presence and absence of 5'-GMP

The reactions of the platinum(II) complexes, [Pt(dien)(H(2)O)](2+), [PtCl(dien)](+) and [PtBr(dien)](+) (dien is diethylenetriamine) with some biologically relevant ligands such as inosine (INO), inosine-5'-monophosphate (5'-IMP), guanosine-5'-monophosphate (5'-GMP), glutathione (GSH) and l-methionine (S-meth), have been studied by UV-Visible spectrophotometry and (1)H NMR spectroscopy. Kinetic and thermodynamic parameters of these reactions were determined. Competitive reactions of [PtCl(dien)](+) with l-methionine and 5'-GMP demonstrated initially rapid formation of [Pt(dien)(S-meth)](2+) followed by displacement of l-methionine by 5'-GMP. In the later stages the concentration of [Pt(dien)(N7-GMP)](2+) is predominant. The results are analyzed in reference to the anti-tumour activity of Pt(II) complexes.     

Serum cholesterol concentrations in suicidal and non-suicidal male patients suffering from persistent delusional disorder

Suicidal behavior is a major health risk in psychiatric disorders, especially in affective and psychotic disorders. The neurobiology of suicidal behavior is still unclear. Suicidality has been related to a reduced cholesterol levels. The aim of this study was to evaluate serum cholesterol concentrations in suicidal and non-suicidal men suffering from persistent delusional disorder and in healthy volunteers. Results showed that serum cholesterol concentrations were significantly lower in suicidal than in non-suicidal patients and healthy controls. Also, level of psychopathology (measured by Brief Psychiatric Rating Scale) is significantly marked in the group of suicidal patients, which indicates the importance of detecting some clinical symptoms in patients with persistent delusional disorder in order to prevent suicidal behavior.     

Variability in blue whale acoustic behavior off southern California

To evaluate the acoustic behavior of blue whales (Balaenoptera musculus) located inshore and offshore of southern California, singular A and B calls, D calls, and AB phrases were analyzed from 12 mo of passive acoustic data collected at four locations within the Southern California Bight. The relative proportions of singular calls and phrases were used to evaluate spatial and temporal patterns in sound and song type usage, and singular call and phrase production rates were calculated to investigate spatial and temporal variability in call abundance. Blue whale sounds were recorded from spring through early winter, with the majority of all detections occurring between September and December. The proportions and production rates of singular calls and phrases varied between the inshore and offshore sites. In addition, the percentage of A units within repetitive song phrases was greater inshore than offshore, resulting from a higher proportion of AB song type inshore, in which A and B phrase units were alternating. The ABB song type, in which a single A unit was followed by multiple B units, was more common offshore. The observed differences in calling and singing behaviors may identify distinct and variable acoustic behavioral settings for blue whales off southern California.     

Localization of carboxypeptidase A-like enzyme in rat kidney

In this study we demonstrate that carboxypeptidase A (CPA)-like enzyme is expressed in rat kidney. The major metabolites of angiotensin (Ang) I by the rat renal mesangial cell extract at 37 degrees C, pH 7.4, were Ang 1-9 and Ang II. Quinaprilat did not influence the formation of Ang 1-9, but it inhibited formation of Ang II. The formation of Ang 1-9 was inhibited by potato carboxypeptidase inhibitor, 1,10-phenanthroline or EDTA. Lowering the pH from 7.4 to 4.0 also inhibited the formation of this nonapeptide. These findings suggest that a metallocarboxypeptidase is responsible for Ang 1-9 production. Using monoclonal antibodies to CPA, Western blot showed the presence of CPA-like enzyme in the extracts prepared from the mesangial cells or kidney cortex of the rat. Immunohistochemistry showed that CPA-like enzyme is localized in the mesangial glomerular cells and adventitia of kidney blood vessels, whereas it was absent in the renal tubules. Our data suggest that a CPA-like enzyme could be added to a repertoire of enzymes present in the rat mesangial cells and adventitia of renal blood vessels.     

Implant failure: regional versus cumulative evaluation

In this paper the success rate of implant therapy in various bone regions is discussed. The objective is to determine whether differences existed in success rates of cylinder implants placed in different areas in the both maxilla and mandible. Forty four patients have been treated and reviewed five years after the placement of the fixed prosthetic restoration. The patients were provided with a total of 92 implants. Results from this study show very low survival rate for implants placed in anterior region of maxilla (55.6%) after five years. It is concluded that simple cumulative follow up studies do not entirely correspond to actual situations, positioning the implants has an important role in the planning of the implant therapy and that important factor for force compensation is not only the surrounding bone density, but also the region of the jaw where the implants are placed.