An improved method for preparing Agrobacterium cells that simplifies the Arabidopsis transformation protocol

BACKGROUND: The Agrobacterium vacuum (Bechtold et al 1993) and floral-dip (Clough and Bent 1998) are very efficient methods for generating transgenic Arabidopsis plants. These methods allow plant transformation without the need for tissue culture. Large volumes of bacterial cultures grown in liquid media are necessary for both of these transformation methods. This limits the number of transformations that can be done at a given time due to the need for expensive large shakers and limited space on them. Additionally, the bacterial colonies derived from solid media necessary for starting these liquid cultures often fail to grow in such large volumes. Therefore the optimum stage of plant material for transformation is often missed and new plant material needs to be grown. RESULTS: To avoid problems associated with large bacterial liquid cultures, we investigated whether bacteria grown on plates are also suitable for plant transformation. We demonstrate here that bacteria grown on plates can be used with similar efficiency for transforming plants even after one week of storage at 4°C. This makes it much easier to synchronize Agrobacterium and plants for transformation. DNA gel blot analysis was carried out on the T(1 )plants surviving the herbicide selection and demonstrated that the surviving plants are indeed transgenic. CONCLUSION: The simplified method works as efficiently as the previously reported protocols and significantly reduces the workload, cost and time. Additionally, the protocol reduces the risk of large scale contaminations involving GMOs. Most importantly, many more independent transformations per day can be performed using this modified protocol

Straight versus S-shaped sternotomy: a histologic study in the sheep model

INTRODUCTION: Straight sternotomy is the most common access for open heart surgery. Techniques have been proposed for maximizing sternal stability in high-risk patients. This trend implies a growing need for newer surgical techniques. The aim of this experimental study in the sheep model is to evaluate median vs. S shaped sternotomy the feasibility of using a special device to accelerate the sternal instability and bone healing. MATERIALS AND METHODS: We enrolled 31 sheep, weighing 18–30 kg. For all animals a midline skin incision was made. In group I (n = 16 animals), straight median sternotomy and in group II (n = 15 animals), S-shaped incision was marked on the periosteum of the sternum by new created device for standard S-shaped sternotomy. Sternum biopsies were performed on second surgery month for all survived animals from the sternum and the surrounding soft tissue. RESULTS: No early superficial wound complications were observed. Overall mortality in the initial approach group was 19.3% (6 sheep). In group I; 3 sheep had died on first surgery day, the reason may be hemorrhage and in group II; 3 sheep developed intractable VF during surgery procedure or immediately afterwards so died. There were statistically significant differences in the scores of bone healing between group 1 and group 2 (4.2 vs.7.3, ANOVA, p < 0.001). DISCUSSION: Our work is based on the use of a standard S-shaped sternotomy procedure on sheep sternum. In our experience with the sternal healing in the sheep model, the process of new bone formation was accelerated with S- shaped cut than straight sternotomy procedure. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13019-014-0173-x) contains supplementary material, which is available to authorized users

Editorial: Green chemistry biocatalysis

Editorial on the Research Topic: Green chemistry biocatalysi

Extractive Dearomatization of Naphthalane Oil Fraction with Ionic Liquid and -N-Methyl-2-Pyrrolidone

In the article are given an analysis of the results of researches carried out for the purpose of selective treatment of the Naphthalane oil fraction boiling at 260-3400C with ionic liquid (IL) -morfolinphormiate synthesized on the basis formic acid + morpholine and - N-methyl-2-pyrrolidon (N-MP) and  as an extractant. The aim is to remove poisonous components -  sulfur compounds and toxic carcinogenic polynuclear aromatic hydrocarbons in order to improve therapeutic properties of Naphthalane oil cut. For this purpose we have used extraction method. On the basis of the conducted researches the role of -N-methyl-2-pyrrolidone and IL-morpholinephormiatee in extraction conditions have been determined in selective treatment of Nafthalan oil, according to the results of spectral analysis residual amount of aromatic hydrocarbons decreased from 18.5% wt accordance to 2%, 1.5%. As well as the amount of sulphur decreased from 0.0354% to 0.010%,  0.011% and 0.019%.In the next stage the treated raffinates with N-methyl-2-pyrrolidone has been purified on silica. In a result amount of aromatic hydrocarbons drastically decrease to be 0.07% and 0.02%. As a result we aquire transparency and at the same time the percentage amount of aromatic hydrocarbons drastically decreased from 2% to 0,07%, 1.5%, to 0.02%. Raffinates obtained in a 2-4 stage dearomatization with N-MP and IL. Raffinate are measured on NMR, UV and IR spectral analysis

The Effects of Two Different Stretching Programs on Balance Control and Motor Neuron Excitability

We examined the effects of training (4d/wk for 6 wks) with static stretching (SS) or contract-relax proprioceptive neuromuscular facilitation (PNF) on static balance time and motor neuron excitability. Static balance time, Hmax/Mmax ratios and H-reflex recovery curves (HRRC) were measured in 28 healthy subjects (SS: n=10, PNF: n=9, control: n=9) before and after training. SS improved static balance time with a trend observed for PNF. Post training, during 150-200-250 msec interstimulus intervals, we observed a reduction in facilitation, but during 500-700-900 msec interstimulus interval; there was an increase in H2/H1 ratio in the PNF group only. Both stretching techniques improved static balance. The Ia afferent inhibitions during the acute exercises were not found after the SS and PNF training programmes. It was concluded that training with contract-relax proprioceptive neuromuscular facilitation may cause some augmentation in supraspinal and postsynaptic inhibition on the motoneuron pool

Evaluation of heart rate variability in patients with coronary artery ectasia and coronary artery disease

Objective: The present study compared heart rate variability (HRV) parameters in patients with coronary artery ectasia (CAE) and coronary artery disease (CAD). Methods: The study population consisted of 60 consecutive patients with CAE (14 women; mean age 51.63±7.44 years), 60 consecutive patients with CA (15 women; mean age 53.67±9.31 years), and 59 healthy individuals (13 women; mean age 52.85±8.19 years). Electrocardiograms, 24-hour Holter analyses, and routine biochemical tests were performed, and clinical characteristics were evaluated. Coronary angiography images were analyzed. Time-domain HRV parameters, including the standard deviation (SD) of normal-to-normal intervals (SDNN) and the root mean square of difference in successive normal-tonormal intervals (RMSSD) were evaluated, as were frequencydomain HRV parameters including low-frequency (LF), very lowfrequency (VLF), high-frequency (HF), the proportion derived by dividing low- and high-frequency (LF/HF), and total power (TP). Results: SDNN was lower in both the CAE and CAD groups, compared to the healthy group (140.85±44.21, 96.51±31.28, and 181.05±48.67, respectively). A significant difference in RMSSD values among the groups was determined (p=0.004). Significantly decreased VLF and HF values were found in the CAE group, compared with the healthy group (VLF p<0.001; HF, p=0.007). TP, VLF, and HF values were significantly lower (p<0.001, p<0.001, and p<0.001, respectively), but LF and LF/ HF values were significantly higher (p<0.001 for both) in the CAD group than in the healthy group. TP values were significantly higher (p<0.001), and LF and LF/HF values were lower in the CAE group, compared with the CAD group (p<0.001 for both). Conclusion: A decrease in vagal modulation or an increase in sympathetic activity of cardiac function, assessed by HRV analysis, is worse in patients with CAD than in patients with CAE. © 2016 Turkish Society of Cardiology

Central defect type partial ACL injury model on goat knees: the effect of infrapatellar fat pad excision

BACKGROUND: The mid-substance central defect injury has been used to investigate the primary healing capacity of the anterior cruciate ligament (ACL) in a goat model. The sagittal plane stability on this model has not been confirmed, and possible effects of fat pad excision on healing have not been evaluated. We hypothesize that excising the fat pad tissue results in poorer ligament healing as assessed histologically and decreased tensile strength of the healing ligament. We further hypothesize that the creation of a central defect does not affect sagittal plane knee stability. METHODS: A mid-substance central defect was created with a 4-mm arthroscopic punch in the ACLs of right knees of all the subjects through a medial mini-arthrotomy. Goats were assigned to groups based on whether the fat pad was preserved (group 1, n = 5) or excised completely (group 2, n = 5). The left knees served as controls in each goat. Histopathology of the defect area along with measurement of type I collagen in one goat from each group were performed at 10th week postoperatively. The remaining knees were evaluated biomechanically at the 12th week, by measuring anterior tibial translation (ATT) of the knee joints at 90° of flexion and testing tensile properties (ultimate tensile load (UTL), ultimate elongation (UE), stiffness (S), failure mode (FM)) of the femur-ACL-tibia complex. RESULTS AND DISCUSSION: Histopathology analysis revealed that the central defect area was fully filled macroscopically and microscopically. However, myxoid degeneration and fibrosis were observed in group 2 and increased collagen type I content was noted in group 2. There were no significant differences within and between groups in terms of ATT values (p = 0.715 and p = 0.149, respectively). There were no significance between or within groups in terms of ultimate tensile load and ultimate elongation; however, group 2 demonstrated greater stiffness than group 1 that was correlated with the fibrotic changes detected microscopically (p = 0.043). CONCLUSIONS: The central defect type injury model was confirmed to be biomechanically stable in a goat model. Resection of the fat pad was noted to negatively affect defect healing and increase ligament stiffness in the central defect injury model

The antimicrobial effects of the alginate oligomer OligoG CF-5/20 are independent of direct bacterial cell membrane disruption

Concerns about acquisition of antibiotic resistance have led to increasing demand for new antimicrobial therapies. OligoG CF-5/20 is an alginate oligosaccharide previously shown to have antimicrobial and antibiotic potentiating activity. We investigated the structural modification of the bacterial cell wall by OligoG CF-5/20 and its effect on membrane permeability. Binding of OligoG CF-5/20 to the bacterial cell surface was demonstrated in Gram-negative bacteria. Permeability assays revealed that OligoG CF-5/20 had virtually no membrane-perturbing effects. Lipopolysaccharide (LPS) surface charge and aggregation were unaltered in the presence of OligoG CF-5/20. Small angle neutron scattering and circular dichroism spectroscopy showed no substantial change to the structure of LPS in the presence of OligoG CF-5/20, however, isothermal titration calorimetry demonstrated a weak calcium-mediated interaction. Metabolomic analysis confirmed no change in cellular metabolic response to a range of osmolytes when treated with OligoG CF-5/20. This data shows that, although weak interactions occur between LPS and OligoG CF-5/20 in the presence of calcium, the antimicrobial effects of OligoG CF-5/20 are not related to the induction of structural alterations in the LPS or cell permeability. These results suggest a novel mechanism of action that may avoid the common route in acquisition of resistance via LPS structural modification