Erratum

Thidiazuron-induced regeneration and genetic transformation of grapevine rootstock varietiesVitis 42 (3), 133- 136 (2003

Thidiazuron-induced regeneration and genetic transformation of grapevine rootstock varieties

In vitro regeneration from cell to plant, a technique considered to be an important precondition of gene transfer, was attempted in 12 grapevine rootstocks starting with anthers of the plants as inocula. Embryogenic callus was induced in solid Murashige-Skoog medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) and thidiazuron (TDZ). Five of the tested rootstock cultivars (Börner, Richter 110, SO 4, St. George and Teleki 5 C) yielded embryogenic callus lines that regenerated entire plants. From these cultures Richter 110 and St. George varieties produced transgenic plants following Agrobacterium-mediated transformation with an nptII/gusA gene construct using a simple, but effective cocultivation protocol. The putative transgenic lines expressed the gusA gene. The T-DNA transfer and integration was confirmed by PCR analysis.

Ultrasound and dynamic functional imaging in vascular cognitive impairment and Alzheimer's disease

The vascular contributions to neurodegeneration and neuroinflammation may be assessed by magnetic resonance imaging (MRI) and ultrasonography (US). This review summarises the methodology for these widely available, safe and relatively low cost tools and analyses recent work highlighting their potential utility as biomarkers for differentiating subtypes of cognitive impairment and dementia, tracking disease progression and evaluating response to treatment in various neurocognitive disorders. METHODS: At the 9th International Congress on Vascular Dementia (Ljubljana, Slovenia, October 2015) a writing group of experts was formed to review the evidence on the utility of US and arterial spin labelling (ASL) as neurophysiological markers of normal ageing, vascular cognitive impairment (VCI) and Alzheimer's disease (AD). Original articles, systematic literature reviews, guidelines and expert opinions published until September 2016 were critically analysed to summarise existing evidence, indicate gaps in current knowledge and, when appropriate, suggest standards of use for the most widely used US and ASL applications. RESULTS: Cerebral hypoperfusion has been linked to cognitive decline either as a risk or an aggravating factor. Hypoperfusion as a consequence of microangiopathy, macroangiopathy or cardiac dysfunction can promote or accelerate neurodegeneration, blood-brain barrier disruption and neuroinflammation. US can evaluate the cerebrovascular tree for pathological structure and functional changes contributing to cerebral hypoperfusion. Microvascular pathology and hypoperfusion at the level of capillaries and small arterioles can also be assessed by ASL, an MRI signal. Despite increasing evidence supporting the utility of these methods in detection of microvascular pathology, cerebral hypoperfusion, neurovascular unit dysfunction and, most importantly, disease progression, incomplete standardisation and missing validated cut-off values limit their use in daily routine. CONCLUSIONS: US and ASL are promising tools with excellent temporal resolution, which will have a significant impact on our understanding of the vascular contributions to VCI and AD and may also be relevant for assessing future prevention and therapeutic strategies for these conditions. Our work provides recommendations regarding the use of non-invasive imaging techniques to investigate the functional consequences of vascular burden in dementia

Evaluation of intron containing potential reference gene-specific primers to validate grapevine nucleic acid samples prepared for conventional PCR and RT-PCR

Previously we proved the usefulness of an intron containing reference gene, phosphoenolpyruvate carboxylase (PEP) to validate cDNA synthesis for detection of grapevine viruses by conventional RT-PCR from crude nucleic acid preparations. Thus amplicons derived from residual genomic DNA (gDNA) and cDNA can be clearly distinguished by their sizes. Here we designed novel sets of primers which encompass one or two intron containing sequences of grapevine housekeeping genes such as actin, tubulin and elongation factor 1-α. Using these primers the expected sequences were amplified from gDNAs of the tested 24 grapevine cultivars. Thereafter they were challenged using cDNAs prepared from total nucleic acid samples isolated from cambial scrapings of dormant canes, leaf laminas, petioles and in vitro leaves of 12 grapevine cultivars. All of these novel, and the previously published PEP gene-specific primers generated the amplification of the expected shorter DNA fragments without introns. Thus they are suitable to check the quality of nucleic acid preparations and to validate subsequent cDNA synthesis prior to pathogen detection assays

Cytotoxic drug sensitivity of Epstein-Barr virus transformed lymphoblastoid B-cells.

BACKGROUND: Epstein-Barr virus (EBV) is the causative agent of immunosuppression associated lymphoproliferations such as post-transplant lymphoproliferative disorder (PTLD), AIDS related immunoblastic lymphomas (ARL) and immunoblastic lymphomas in X-linked lymphoproliferative syndrome (XLP). The reported overall mortality for PTLD often exceeds 50%. Reducing the immunosuppression in recipients of solid organ transplants (SOT) or using highly active antiretroviral therapy in AIDS patients leads to complete remission in 23-50% of the PTLD/ARL cases but will not suffice for recipients of bone marrow grafts. An additional therapeutic alternative is the treatment with anti-CD20 antibodies (Rituximab) or EBV-specific cytotoxic T-cells. Chemotherapy is used for the non-responding cases only as the second or third line of treatment. The most frequently used chemotherapy regimens originate from the non-Hodgkin lymphoma protocols and there are no cytotoxic drugs that have been specifically selected against EBV induced lymphoproliferative disorders. METHODS: As lymphoblastoid cell lines (LCLs) are well established in vitro models for PTLD, we have assessed 17 LCLs for cytotoxic drug sensitivity. After three days of incubation, live and dead cells were differentially stained using fluorescent dyes. The precise numbers of live and dead cells were determined using a custom designed automated laser confocal fluorescent microscope. RESULTS: Independently of their origin, LCLs showed very similar drug sensitivity patterns against 29 frequently used cytostatic drugs. LCLs were highly sensitive for vincristine, methotrexate, epirubicin and paclitaxel. CONCLUSION: Our data shows that the inclusion of epirubicin and paclitaxel into chemotherapy protocols against PTLD may be justified

Spectroscopic and photometric confirmation of chromospheric activity in four stars

We present analysis of medium resolution optical spectra and long term V band photometry of four cool stars, BD+13 5000, BD+11 3024, TYC 3557-919-1 and TYC 5163-1764-1. Our spectroscopic analysis reveals that the stars are giant or sub-giant from K0 or K1 spectral type, and all of them exhibit emission features in their Ca ii H& K lines. These features appear to be modulated with the rotation of the stars. Except BD+11 3024, we observe that the radial velocities of the target stars are not stable, which suggests that each of them might be a member of a binary system. Global analysis of photometric data indicates clear cyclic variation for BD+13 5000 and TYC 5163-1764-1 with a period of 8.0$\pm$0.3 and 5.04$\pm$0.04 year, respectively. Besides that, we observe a dramatic increase ($\sim$0\fm7) in the mean brightness of BD+11 3024, accompanied with a 2.87$\pm$0.12 cyclic variation, embedded into the global brightening trend, which indicates possible multiple cycles on this star.Comment: 10 pages, 6 figures, 5 tables, accepted for publication in A

An improved method for embryogenic suspension cultures of 'Richter 110' rootstock

Research Not