Identification and expression analyses of poly [I:C]-stimulated genes in channel catfish (Ictalurus punctatus)

Channel catfish (Ictalurus punctatus) have proven to be an excellent model with which to study immune responses of lower vertebrates. Identification of anti-viral antibodies and cytotoxic cells, as well as both type I and II interferon (IFN), demonstrates that catfish likely mount a vigorous anti-viral immune response. In this report, we focus on other elements of the anti-viral response, and identify more than two dozen genes that are induced following treatment of catfish cells with poly [I:C]. We showed that poly [I:C] induced type I interferon within 2 h of treatment, and that characteristic interferon stimulated genes (ISGs) appeared 6–12 h after exposure. Among the ISGs detected by RT-PCR assay were homologs of ISG15, Mx1, IFN regulatory factor 1 (IRF-1), inhibitor of apoptosis protein-1 (IAP-1) and the chemokine CXCL10. Microarray analyses showed that 13 and 24 cellular genes, respectively, were upregulated in poly [I:C]-treated B cell and fibroblast cultures. Although many of these genes were novel and did not fit the profile of mammalian ISGs, there were several (ISG-15, ubiquitin-conjugating enzyme E2G1, integrin-linked kinase, and clathrin-associated protein 47) that were identified as ISGs in mammalian systems. Taken together, these results suggest that dsRNA, either directly or through the prior induction of IFN, upregulates catfish gene products that function individually and/or collectively to inhibit virus replication.     

Response of barley seedlings to UV-B radiation as affected by NaCl

The response of barley seedlings, subjected to 150 mmol/L NaCl for 4 days at different light regimes (4 d in the light, 4 d in darkness and a 12 h light/dark cycle) before UV-B radiation was investigated. NaCl treatment resulted in a decrease of total chlorophyll content and an increase in H2O2, free proline and lipid peroxidation, as quantified by measurement of malondialdehyde. Significantly more proline was accumulated in the light than in darkness. The combination of UV-B and NaCl treatment produced an additive effect on most of the parameters studied. UV-B radiation reduced the chlorophyll/carotenoids ratio and photochemical efficiency of PSII as estimated by chlorophyll fluorescence. NaCl pre-exposure decreased H2O2 generation and lipid peroxidation and alleviated the inhibitory effect of UV-B on PSII activity. Proline accumulated under salt stress conditions might be one of the reasons for the observed tolerance of barley seedlings to UV-B radiation.     

Localization of ligand-binding sites on human C1q globular head region using recombinant globular head fragments and single-chain antibodies

As a charge pattern recognition molecule, human C1q can bind a range of immunoglobulin and non-immunoglobulin ligands via its carboxy-terminal globular domain and activate the classical complement pathway. Each globular domain has a heterotrimeric organization, composed of the carboxy-terminal halves of one A (ghA), one B (ghB), and one C (ghC) chain. Recently, we have found that the recombinant forms of individual ghA, ghB and ghC bind differentially to IgG, IgM, gp41 peptide 601-613 of human immunodeficiency virus-1 (HIV-1), gp21 peptide 400-429 of human T cell lymphotrophic virus-I (HTLV-I), beta-amyloid peptide, and apoptotic cells, suggesting a modular organization of the globular domain. This paper examines the interaction of ghA, ghB and ghC with two known C1q ligands: Klebsiella pneumoniae porin OmpK36 and salivary agglutinin. In addition, we have used a panel of recombinant single-chain antibodies (scFv) specific for ghA, ghB and ghC in order to map sites on the heterotrimeric globular domain which are likely to interact with IgG1, IgG3, IgM, OmpK36, salivary agglutinin and gp41 loop peptide. The combined use of recombinant ghA, ghB, ghC and single-chain antibodies has revealed at least three ligand-binding sites on the globular domain of C1q: one is IgG- and OmpK36-specific, the second (IgM-binding site) is most likely overlapping with IgG/OmpK36 binding site, and the third (the gp41-binding site) seems to be located at the junction between the collagen and globular domains.     

Phacoemulsification and silicone oil removal through the planned posterior capsulorhexis

The purpose of the study was to present operative technique and results of a passive hydrodynamic expression of silicone oil through planned posterior capsulorhexis during cataract surgery in patients after pars plana vitrectomy. The retrospective analysis was done on 57 eyes with cataract after a previous pars plana vitrectomy, operated on between 2001 and 2004 at the Clinical hospital "Sestre milosrdnice" Zagreb. Preoperative and postoperative best corrected visual acuity (BCVA), preoperative and postoperative intraocular pressure (IOP), and postoperative complications were reviewed. Visual acuity improved or stabilized in all patients with an attached retina. Retinal detachment occurred in 11 eyes. Transient vitreous hemorrhage, that resolved within 1 week of surgery without treatment, was observed in 4 eyes. Asymptomatic intraocular lens (IOL) decentration occurred in 2 eyes. Our findings suggest that silicone oil removal and cataract surgery can be performed as a single procedure in selected patients in the absence of macular pucker and retinal reproliferation, and in a presence of a stable retina.     

Postoperative outcomes after implantation of intraocular lenses in eyes with cataract and uveitis

Despite advances in surgical technique and implant materials, cataract surgery in patients with uveitis is still a challenging procedure. We retrospectively evaluated postoperative outcomes of cataract surgery in 35 eyes of 29 patients with uveitis. Phacoemulsification with posterior chamber intraocular lens implantation was performed in all eyes. Postoperative evaluations were performed at day 2, and then at 7 days, 1, 3, and 6 months respectively. There were 16 males, and 13 females, aged 31 to 68 years. Follow-up ranged from 4 to 35 months. At final follow-up 33 eyes (94%) had an improvement in visual acuity compared with preoperative levels (p < 0,05). Giant cells were observed in the intraocular lens optic in 7 eyes (20%). Posterior capsule opacification occurred in 10 eyes (29%). Clinical cystoid macular edema was observed in 4 eyes, and 2 eyes required trabeculectomy with mitomycin C due to secondary glaucoma. Cataract surgery in patients with uveitis leads to successful visual results after correct surgical timing, and adequate anti-inflammatory therapy. There were no significant differences in the degree of inflammation after implantation of various types of intraocular lenses.     

Spread and competitiveness of Plum Pox Virus: Rec and ‐D strains in experimental plum orchard

Sharka disease caused by Plum pox virus (PPV) is endemic in Bulgaria. The speed and mode of spread of PPV‐Rec and PPV‐D isolates, molecularly characterised in advance and introduced in an experimental plum orchard to serve as permanent sources of infection was studied from 2008 to 2015. The trees were visually inspected for symptoms development and tested by DASI‐ELISA twice each year. The positive samples were serologically (‐M and ‐D specific MAbs) and molecularly (IC‐RT‐PCR) strain‐typed, and sequenced in (Cter)NIb‐(Nter)CP coding region. Artificially inoculated trees were found infected in the spring of 2009. After 7 years 43 trees (11.2%) became naturally infected. The majority of the trees were infected by PPV‐Rec (65%), while only one tree (2.5%) by PPV‐D. Nevertheless PPV‐M was not introduced in the orchard, this strain was found in single and mixed infections with PPV‐Rec strain. None of the isolates in naturally infected trees was identical to PPV‐Rec and PPV‐D inoculums, while several of them were a 100% identical to the respective PPV‐Rec, PPV‐D or PPV‐M isolates from neighbourhood‐infected trees. After the initial random pattern of PPV spread, aggregation of the new infections around the trees infected in previous years was observed. The presence of only Phorodon humuli and Hyalopterus pruni complex, known as PPV vectors and their poor abundance (5.0% from all caught aphids), together with some specific features of the landscape influenced the slow natural development of PPV in the experimental orchard.     

GC-mS analysis and anti-microbial activity of acidic fractions obtained from Paeonia peregrina and Paeonia tenuifolia roots

Fourteen aromatic and 24 aliphatic acids were determined by GC-MS analysis of acidic fractions obtained from Paeonia peregrina and Paeonia tenuifolia roots. Benzoic acid and its monohydroxy-, dihydroxy- and trihydroxy-derivatives are the main acid components of both Paeonia species. Some fractions could serve as a source of benzoic, 4-hydroxybenzoic, vanillic and gallic acids, as well as of ethyl gallate. The fractions inhibited the growth of Staphylococcus aureus, Escherichia coli and Candida albicans.     

Synthesis and anti-enterovirus activity of new analogues of MDL-860

A series of twelve novel compounds, analogues of antiviral agent MDL-860 were synthesized and their antiviral activity was evaluated in vitro against enteroviruses poliovirus 1 (PV1), Coxsackieviruses B1 (CVB1) and Coxsackieviruses B3 (CVB3). Compounds 14, 24 and 25 manifested strong antiviral effects against CVB1 and PV1 (SI values of 405 and 118 for CVB1 and PV1 respectively). In contrast to the wide anti-enteroviral activity of MDL-860, these three compounds were inactive against CVB3. Compounds 14, 24 and 25 along with MDL-860 were tested in vivo in mice infected with CVB1. Marked protective effects of compounds 14 and 24 were established, PI values of 50% and 33.3%, respectively. In addition, almost all of the tested compounds manifested very low toxicity.