A short version of the reflective functioning questionnaire: Validation in a greek sample.

This study aims to validate the Greek version of the 54-item Reflective Functioning Questionnaire (RFQ), a measure designed to assess an individual's capacity for understanding themselves and others based on internal mental states. This capacity, also known as Reflective Functioning (RF) or mentalizing, is believed to play a significant role in both typical and atypical development. The validation process examined the factor structure of the RFQ and its relationship with a variety of psychosocial and clinical constructs that have theoretical and empirical links to RF. Additionally, this research investigated the factor structure's invariance across gender and age groups to determine the robustness of the instrument. A unique contribution of this work lies in examining the application of the RFQ to attachment classifications through the use of cluster analysis. The sample consisted of 875 Greek adults from the general community with a mean age of 28.5 and a median age of 22. Participants completed the Greek RFQ along with a series of self-report questionnaires assessing psychosocial constructs, including attachment, epistemic trust, emotion regulation, and psychological mindedness, as well as clinical variables such as anxiety, depression, and borderline personality traits. Our findings suggest that a shorter, 31-item version of the questionnaire provides a robust three-factor structure across a non-clinical Greek adult population. The three identified subscales are (a) excessive certainty, (b) interest/curiosity, and (c) uncertainty/confusion, all demonstrating satisfactory reliability and construct validity. The uncertainty subscale was found to be associated with insecure attachment styles, epistemic mistrust and credulity, emotional suppression, and low psychological mindedness. In contrast, the certainty and curiosity subscales were linked to secure attachment, epistemic trust, emotion reappraisal, and psychological mindedness. Uncertainty was further shown to differ significantly across probable clinical and non-clinical groups, as distinguished by cut-off scores for anxiety, depression, and borderline personality disorder (BPD). However, the certainty and interest/curiosity subscales only varied between the two BPD groups. Our results provide the first evidence supporting the use of a 31-item version of the RFQ with three validated subscales to reliably assess reflective functioning in the Greek population, demonstrating stronger psychometric properties compared to other RFQ versions reported in previous studies. Findings suggest that impaired mentalizing capacity, as measured by the RFQ, is linked to insecure attachment, epistemic mistrust and credulity, poor emotion regulation, and low psychological mindedness, and potentially plays a role in adult mental health symptoms.

Solid-Phase Synthesis of 2-Benzothiazolyl and 2-(Aminophenyl)benzothiazolyl Amino Acids and Peptides.

2-benzothiazoles and 2-(aminophenyl)benzothiazoles represent biologically interesting heterocycles with high pharmacological activity. The combination of these heterocycles with amino acids and peptides is of special interest, as such structures combine the advantages of amino acids and peptides with the advantages of the 2-benzothiazolyl and 2-(aminophenyl)benzothiazolyl pharmacophore group. In this work, we developed an easy and efficient method for the solid-phase synthesis of 2-benzothiazolyl (BTH) and 2-(aminophenyl)benzothiazolyl (AP-BTH) C-terminal modified amino acids and peptides with high chiral purity.

Ion exchange chromatographic conditions for obtaining individual subunits of soybean beta-conglycinin.

Soybean beta-conglycinin is a complex protein possessing health-promoting properties. beta-Conglycinin is a trimeric glycoprotein. Little information related to methods for separation of the individual chains forming beta-conglycinin has been so far published and it is of great interest. As a consequence, less data on the bioactivities of alpha, alpha' and beta subunits of this glycoprotein have been published. The present research aimed to find out new alternative chromatographic conditions to obtain beta-conglycinin subunits that are free of contaminating proteins. In the present short communication, we propose the use of a two-step ion exchange chromatographic protocol to achieve this goal. Firstly, beta subunit was separated by means of anionic exchange fast protein liquid chromatography. Secondly, alpha and alpha' chains were separated from each other by cationic exchange. Our data indicated the feasibility of proposed fractionation protocol to separate soybean beta-conglycinin alpha and alpha' subunits from other contaminating proteins and to obtain enough amounts of the three individual chains forming this glycoprotein for further characterization and application. The procedure may be easily up-scaled.

The production, purification and characterisation of two novel alpha-D-mannosidases from Aspergillus phoenicis.

1,6-alpha-D-Mannosidase from Aspergillus phoenicis was purified by anion-exchange chromatography, chromatofocussing and size-exclusion chromatography. The apparent molecular weight was 74 kDa by SDS-PAGE and 81 kDa by native-PAGE. The isoelectric point was 4.6. 1,6-alpha-D-Mannosidase had a temperature optimum of 60 degrees C, a pH optimum of 4.0-4.5, a K(m) of 14 mM with alpha-D-Manp-(1-->6)-D-Manp as substrate. It was strongly inhibited by Mn(2+) and did not need Ca(2+) or any other metal cofactor of those tested. The enzyme cleaves specifically (1-->6)-linked mannobiose and has no activity towards any other linkages, p-nitrophenyl-alpha-D-mannopyranoside or baker's yeast mannan. 1,3(1,6)-alpha-D-Mannosidase from A. phoenicis was purified by anion-exchange chromatography, chromatofocussing and size-exclusion chromatography. The apparent molecular weight was 97 kDa by SDS-PAGE and 110 kDa by native-PAGE. The 1,3(1,6)-alpha-D-mannosidase enzyme existed as two charge isomers or isoforms. The isoelectric points of these were 4.3 and 4.8 by isoelectric focussing. It cleaves alpha-D-Manp-(1-->3)-D-Manp 10 times faster than alpha-D-Manp-(1-->6)-D-Manp, has very low activity towards p-nitrophenyl-alpha-D-mannopyranoside and baker's yeast mannan, and no activity towards alpha-D-Manp-(1-->2)-D-Manp. The activity towards (1-->3)-linked mannobiose is strongly activated by 1mM Ca(2+) and inhibited by 10mM EDTA, while (1-->6)-activity is unaffected, indicating that the two activities may be associated with different polypeptides. It is also possible that one polypeptide may have two active sites catalysing distinct activities.

Synthesis of galacto-oligosaccharide from lactose using beta-galactosidase from Kluyveromyces lactis: Studies on batch and continuous UF membrane-fitted bioreactors.

A study of galacto-oligosaccharides (GOS) synthesis from lactose with beta-galactosidase from Kluyveromyces lactis (Maxilact L2000) was carried out. The synthesis was performed using various initial lactose concentrations ranging from 220 to 400 mg/mL and enzyme concentrations ranging from 3 to 9 U/mL, and was investigated at 40 degrees C and pH 7, in a stirred-tank reactor. In the experimental range examined, the results showed the amount of GOS formed depended on lactose concentration but not on enzyme concentration. Galactose was a competitive inhibitor, while glucose was a non-competitive inhibitor. In a further study, a laboratory-scale reactor system, fitted with a 10-kDa NMWCO composite regenerated cellulose membrane, was used in a continuous process. The reactor was operated in cross-flow mode. The effect of operating pressures on flux and productivity was investigated by applying different transmembrane pressures to the system. The continuous process showed better production performance compared to the batch synthesis with the same lactose and enzyme concentrations at 40 degrees C, pH 7. Comparison of product structures from batch and continuous processes, analyzed by HPAE-PAD and methylation analysis, showed similarities but differed from the structures found in a commercial GOS product (Vivinal GOS).