Antitumor Effects of Recombinant Antivascular Protein ABRaA-VEGF121 Combined with IL-12 Gene Therapy

Development and neoplastic progression strongly rely on tumor microenvironment cells. Various kinds of cells that form such tumor milieu play substantial roles in angiogenesis and immunosuppression. Attempts to inhibit tumor vascularization alter tumor milieu and enhance immune response against the tumor. Anticancer therapeutic strategy bringing together antiangiogenic and immunostimulating agents has emerged as a promising approach. We here investigated whether therapy directed against preexisting vessels, combined with an immunomodulatory factor would be equally effective in arresting tumor growth. To this goal, we investigated the effectiveness of ABRaA-vascular endothelial growth factor isoform 121 (VEGF₁₂₁), an antivascular drug constructed by us. It is a fusion protein composed of VEGF₁₂₁, and abrin A chain (translation-inhibiting toxin). We used it in combination with interleukin (IL-12) gene therapy and tried to inhibit B16-F10 melanoma tumor growth. ABRaA-VEGF₁₂₁ is a chimeric recombinant protein capable of destroying tumor vasculature and triggering necrosis in the vicinity of damaged vessels. IL-12 cytokine, in turn, activates both specific and non-specific immune responses. Our results demonstrate that combination of ABRaA-VEGF₁₂₁ antivascular agent with immunostimulatory cytokine IL-12 indeed inhibits tumor growth more effectively than either agent alone, leading to complete cure of ca. 20 % mice. Post-therapeutic analysis of tumors excised from mice treated with combination therapy showed decreased numbers of blood microvessels in the tumor microenvironment, lowered numbers of regulatory T lymphocytes, as well as showed higher levels of CD4⁺ and CD8⁺ as compared to control mice. It seems that bringing together antivascular strategy and the action of immunostimulating agents indeed inhibits growth of tumors.     

Diagnostic validity Polish language version of the questionnaire MINI-KID (Mini International Neuropsychiatry Interview for Children and Adolescent)

Objective

Since over forty years structuralized interviews for clinical and epidemiological research in child and adolescent psychiatry are being developed that should increase validity and reliability of diagnoses according to classification systems (DSM and ICD). The aim of the study is to assess the validity of the Polish version of MINI-KID (Mini International Neuropsychiatric Interview for Children and Adolescents) in comparison to clinical diagnosis made by a specialist in the field of child and adolescent psychiatry.

Materials and methods

There were 140 patients included in the study (93 boys, 66.4%, mean age 11.8 ± 3.0 and 47 girls 33.5%, mean age 14.0 ± 2.9). All the patients were diagnosed by the specialist in the field of child and adolescent psychiatry according to ICD-10 criteria and by the independent interviewer with the Polish version of MINI-KID (version 2.0, 2001).

Results

There was higher agreement between clinical diagnoses and diagnoses based on MINI-KID interview with respect to eating disorders and externalizing disorders (κ 0.43–0.56) and lower in internalizing disorders (κ 0.13–0.45). In the clinical interview, there was smaller number of diagnostic categories (maximum 3 diagnoses per one patient) in comparison to MINI-KID (maximum 10 diagnoses per one patient), and the smaller percentage of patients with one diagnosis (65,7%) in comparison to MINI-KID interview (72%).

Conclusion

Our study has shown satisfactory validity parameters of MINI-KID questionnaire, promoting its use for clinical and epidemiological settings.

Implications and contribution

The Mini International Neuropsychiatry Interview for Children and Adolescent (MINI-KID) is the first structuralized diagnostic interview for assessing mental status in children and adolescents, which has been translated into Polish language. Our validation study demonstrated satisfactory psychometric properties of the questionnaire, enabling its use in clinical practice and in research projects.     

Acute coronary syndrome during dissection of left main as a complication of radiofrequency ablation

We present a case of 44 year-old female who was admitted to the hospital due to performed radio frequency ablation because of VF during WPW syndrome, which was complicated by dissection of left main. The dissection was treated with success by primary percutaneous coronary intervention with two metal stents.     

Rotational angiography in monitoring of covered CP stent implantation in patient with critical aortic coarctation and patent ductus arteriosus

In presented case rotational angiography with three dimensional reconstruction (3DRA) was used for diagnostic and control angiograms during covered CP stent implantation in patient with critical aortic coarctation and patent ductus arteriosus. Administering less contrast then for standard two perpendicular projections, good quality images were obtained in at least seven projections.     

Monomeric site-specific nucleases for genome editing

Targeted manipulation of complex genomes often requires the introduction of a double-strand break at defined locations by site-specific DNA endonucleases. Here, we describe a monomeric nuclease domain derived from GIY-YIG homing endonucleases for genome-editing applications. Fusion of the GIY-YIG nuclease domain to three-member zinc-finger DNA binding domains generated chimeric GIY-zinc finger endonucleases (GIY-ZFEs). Significantly, the I-TevI-derived fusions (Tev-ZFEs) function in vitro as monomers to introduce a double-strand break, and discriminate in vitro and in bacterial and yeast assays against substrates lacking a preferred 5'-CNNNG-3' cleavage motif. The Tev-ZFEs function to induce recombination in a yeast-based assay with activity on par with a homodimeric Zif268 zinc-finger nuclease. We also fused the I-TevI nuclease domain to a catalytically inactive LADGLIDADG homing endonuclease (LHE) scaffold. The monomeric Tev-LHEs are active in vivo and similarly discriminate against substrates lacking the 5'-CNNNG-3' motif. The monomeric Tev-ZFEs and Tev-LHEs are distinct from the FokI-derived zinc-finger nuclease and TAL effector nuclease platforms as the GIY-YIG domain alleviates the requirement to design two nuclease fusions to target a given sequence, highlighting the diversity of nuclease domains with distinctive biochemical properties suitable for genome-editing applications.