Genetic and protein sequence variation of CBF1-4 in cold hardy wild grapevine germplasm

Grapevine (Vitis vinifera) production is limited by a wide array of abiotic stresses including cold, drought, and salinity stresses. The regulation of stress response genes under these conditions is very complex but a common and essential component of the gene network is the C-Repeat Binding Factor (CBF) genes. Four unique CBF genes have been described in grapevine to date and gene or protein level variation may help explain differences in stress resistance phenotypes in grapevine. This study was conducted to evaluate the level of genetic diversity found at these genes in different wild grapevine species as well as in a panel of cultivated grapevine varieties. Examination of the genetic variation at the four CBF genes in grapevine revealed high sequence variation with many different alleles in wild and cultivated grape. However, predicted protein sequence variation of the different genes revealed a different pattern. Vitis CBF genes 1, 2, and 3, which play roles in the stress response to many different stresses, were seen to have high levels of protein sequence variation. In contrast, VCBF4, which is induced by cold, was seen to have very little variation across all wild and cultivated grapevine samples. This contrasting pattern between different gene family members suggests an essential role of VCBF4 in grapevine as the protein sequence is highly conserved between wild North American grapevine species and cultivated varieties.

Influences of obese (ob/ob) and diabetes (db/db) genotype mutations on lumber vertebral radiological and morphometric indices: Skeletal deformation associated with dysregulated systemic glucometabolism

BACKGROUND: Both diabetes and obesity syndromes are recognized to promote lumbar vertebral instability, premature osteodegeneration, exacerbate progressive osteoporosis and increase the propensity towards vertebral degeneration, instability and deformation in humans. METHODS: The influences of single-gene missense mutations, expressing either diabetes (db/db) or obese (ob/ob) metabolic syndromes on vertebral maturation and development in C57BL/KsJ mice were evaluated by radiological and macro-morphometric analysis of the resulting variances in osteodevelopment indices relative to control parameters between 8 and 16 weeks of age (syndrome onset @ 4 weeks), and the influences of low-dose 17-B-estradiol therapy on vertebral growth expression evaluated. RESULTS: Associated with the indicative genotypic obesity and hyper-glycemic/-insulinemic states, both db/db and ob/ob mutants demonstrated a significant (P ≤ 0.05) elongation of total lumbar vertebrae column (VC) regional length, and individual lumbar vertebrae (LV1-5) lengths, relative to control VC and LV parameters. In contrast, LV1-5 width indices were suppressed in db/db and ob/ob mutants relative to control LV growth rates. Between 8 and 16 weeks of age, the suppressed LV1-5 width indices were sustained in both genotype mutant groups relative to control osteomaturation rates. The severity of LV1-5 width osteosuppression correlated with the severe systemic hyperglycemic and hypertriglyceridemic conditions sustained in ob/ob and db/db mutants. Low-dose 17-B-estradiol therapy (E2-HRx: 1.0 ug/ 0.1 ml oil s.c/3.5 days), initiated at 4 weeks of age (i.e., initial onset phase of db/db and ob/ob expressions) re-established control LV 1–5 width indices without influencing VC or LV lengths in db/db groups. CONCLUSION: These data demonstrate that the abnormal systemic endometabolic states associated with the expression of db/db and ob/ob genomutation syndromes suppress LV 1–5 width osteomaturation rates, but enhanced development related VC and LV length expression, relative to control indices in a progressive manner similar to recognized human metabolic syndrome conditions. Therapeutic E2 modulation of the hyperglycemic component of diabetes-obesity syndrome protected the regional LV from the mutation-induced osteopenic width-growth suppression. These data suggest that these genotype mutation models may prove valuable for the evaluation of therapeutic methodologies suitable for the treatment of human diabetes- or obesity-influenced, LV degeneration-linked human conditions, which demonstrate amelioration from conventional replacement therapies following diagnosis of systemic syndrome-induced LV osteomaturation-associated deformations

PII: S0166-2236(99)01425-3

Recent experimental findings show that fast synaptic transmission can extend its actions beyond the immediate synaptic cleft.Whether this phenomenon results in significant crosstalk between typical neighbouring synapses remains unclear. This article considers two areas of the hippocampus, the CA1 and dentate gyrus, where important neural processing occurs.The results discussed do not provide a simple answer to the question of whether synapses can 'talk' to their neighbours, but they do reveal crucial physiological constraints that determine the significance of synaptic crosstalk, thus adding considerably to our understanding of chemical synaptic transmission

Phase II Trial of IL-12 Plasmid Transfection and PD-1 Blockade in Immunologically Quiescent Melanoma.

PurposeTumors with low frequencies of checkpoint positive tumor-infiltrating lymphocytes (cpTIL) have a low likelihood of response to PD-1 blockade. We conducted a prospective multicenter phase II trial of intratumoral plasmid IL-12 (tavokinogene telseplasmid; "tavo") electroporation combined with pembrolizumab in patients with advanced melanoma with low frequencies of checkpoint positive cytotoxic lymphocytes (cpCTL).Patients and methodsTavo was administered intratumorally days 1, 5, and 8 every 6 weeks while pembrolizumab (200 mg, i.v.) was administered every 3 weeks. The primary endpoint was objective response rate (ORR) by RECIST, secondary endpoints included duration of response, overall survival and progression-free survival. Toxicity was evaluated by the CTCAE v4. Extensive correlative analysis was done.ResultsThe combination of tavo and pembrolizumab was well tolerated with adverse events similar to those previously reported with pembrolizumab alone. Patients had a 41% ORR (n = 22, RECIST 1.1) with 36% complete responses. Correlative analysis showed that the combination enhanced immune infiltration and sustained the IL-12/IFNγ feed-forward cycle, driving intratumoral cross-presenting dendritic cell subsets with increased TILs, emerging T cell receptor clones and, ultimately, systemic cellular immune responses.ConclusionsThe combination of tavo and pembrolizumab was associated with a higher than expected response rate in this poorly immunogenic population. No new or unexpected toxicities were observed. Correlative analysis showed T cell infiltration with enhanced immunity paralleling the clinical activity in low cpCTL tumors

An Emerging Model of Creative Game-based Learning

We consider the integration of creative approaches to problem solving into pervasive games is a natural extension of play for creative thinking – one that can innovatively drive technology-led changes to the facilitation of creative thinking and pose a new genre in serious gaming for learning. This paper presents an initial proposal of a new model of creative game-base learning (CGBL), which emerged through mapping of established characteristics of climates that encourage creativity and innovation to characteristics of effective serious games

Chilling requirements and dormancy evolution in grapevine buds.

Fluctuations in winter chilling availability impact bud dormancy and budburst. The objective of this work was to determine chilling requirements to induce and overcome endodormancy (dormancy controlled by chilling) of buds in different grape cultivars. "Chardonnay", "Merlot" and "Cabernet Sauvignon" shoots were collected in Veranópolis-RS vineyards in 2010, and submitted to a constant 3 °C temperature or daily cycles of 3/15 °C for 12/12h or 18/6h, until reaching 1120 chilling hours (CH, sum of hours with temperature ≤ 7.2 °C). Periodically, part of the samples in each treatment was transferred to 25 °C for budburst evaluation (green tip). Chilling requirements to induce and overcome endodormancy vary among cultivars, reaching a total of 136 CH for "Chardonnay", 298 CH for "Merlot" and 392 CH for "Cabernet Sauvignon". Of these, approximately 39, 53 and 91 CH are required for induction of endodormancy in the three cultivars, respectively. The thermal regimes tested (constant or alternating) do not influence the response pattern of each cultivar to cold, with 15 °C being inert in the CH accumulation process. In addition, time required to start budburst reduces with the increase in CH, at a rate of one day per 62 CH, without significant impacts on budburst uniformity. Index terms: Chilling hours; endodormancy; budburst; Vitis vinifera