A method for determining the apoplastic water volume of conifer needles

A method for direct estimation of percentage apoplastic water volume (% APO) in conifer needles is described. The method presented here, and designated the pressure-needle (P-N) method, measures the relative water content of the needles to develop a curve similar to the pressure-volume (P-V) curve. P-V and P-N curves were developed for Picea pungens Engelm. cv. Hoopsi, Pinus sylvestris L., Abies gradis (Dougl.) L., and Pseudotsuga menziesii (Mirb) Franco. The % APO estimated by the two procedures varied as much as 2-fold, while other parameters were similar. The P-V method generated consistently higher and more variable % APO than the P-N method, due to the inclusion of the apoplastic water of the stem in the P-V method. For conifers, the P-N method offers a more accurate and precise method for determining % APO.     

Using matrix summation method for three dimensional dose calculation in brachytherapy

AimThe purpose of this study is to calculate radiation dose around a brachytherapy source in a water phantom for different seed locations or rotation the sources by the matrix summation method.BackgroundMonte Carlo based codes like MCNP are widely used for performing radiation transport calculations and dose evaluation in brachytherapy. But for complicated situations, like using more than one source, moving or rotating the source, the routine Monte Carlo method for dose calculation needs a long time running.Materials and methodsThe MCNPX code has been used to calculate radiation dose around a ¹⁹²Ir brachytherapy source and saved in a 3D matrix. Then, we used this matrix to evaluate the absorbed dose in any point due to some sources or a source which shifted or rotated in some places by the matrix summation method.ResultsThree dimensional (3D) dose results and isodose curves were presented for ¹⁹²Ir source in a water cube phantom shifted for 10 steps and rotated for 45 and 90° based on the matrix summation method. Also, we applied this method for some arrays of sources.ConclusionThe matrix summation method can be used for 3D dose calculations for any brachytherapy source which has moved or rotated. This simple method is very fast compared to routine Monte Carlo based methods. In addition, it can be applied for dose optimization study.     

Multi-parametric analysis and modeling of relationships between mitochondrial morphology and apoptosis

Mitochondria exist as a network of interconnected organelles undergoing constant fission and fusion. Current approaches to study mitochondrial morphology are limited by low data sampling coupled with manual identification and classification of complex morphological phenotypes. Here we propose an integrated mechanistic and data-driven modeling approach to analyze heterogeneous, quantified datasets and infer relations between mitochondrial morphology and apoptotic events. We initially performed high-content, multi-parametric measurements of mitochondrial morphological, apoptotic, and energetic states by high-resolution imaging of human breast carcinoma MCF-7 cells. Subsequently, decision tree-based analysis was used to automatically classify networked, fragmented, and swollen mitochondrial subpopulations, at the single-cell level and within cell populations. Our results revealed subtle but significant differences in morphology class distributions in response to various apoptotic stimuli. Furthermore, key mitochondrial functional parameters including mitochondrial membrane potential and Bax activation, were measured under matched conditions. Data-driven fuzzy logic modeling was used to explore the non-linear relationships between mitochondrial morphology and apoptotic signaling, combining morphological and functional data as a single model. Modeling results are in accordance with previous studies, where Bax regulates mitochondrial fragmentation, and mitochondrial morphology influences mitochondrial membrane potential. In summary, we established and validated a platform for mitochondrial morphological and functional analysis that can be readily extended with additional datasets. We further discuss the benefits of a flexible systematic approach for elucidating specific and general relationships between mitochondrial morphology and apoptosis.     

Advances in brachytherapy

During the past 15 years, permanent seed brachytherapy for prostate cancer has advanced significantly in all areas, including patient selection, treatment planning, technique, and technology (eg, seeds stranded in Vicryl suture). These improvements have made transperineal seed implantation an accurate and practical treatment option for men with low-, intermediate-, and high-risk disease. Because of the evidence that the various treatment options for prostate cancer are likely to be equally successful in terms of long-term cancer control, continuing efforts focus on enhancing the quality of life of implant patients.     

Real-time PCR, a method fit for detection and quantification of Erwinia amylovora

Fire blight, a devastating disease of pome fruit trees continues to pose threat to agricultural production. Detection of its causative agent, bacterium Erwinia amylovora, is usually straightforward in symptomatic samples. Methods with increased sensitivity however, are sometimes needed for detection of E. amylovora and real-time PCR assays have been shown to have required sensitivity and reliability. Here we summarize our previous results on real-time PCR detection of fire blight and present new, fast and sensitive real-time PCR assay based on amsC gene performed on SmartCycler^? instrument. The setting is optimal for analysis of small number of samples in the laboratory or for on-site detection. Many advantages of real-time PCR assays warrant their use in detection and diagnosis of E. amylovora, particularly in detection of low concentrations of target bacteria e.g. in testing for latent infections. It is to be expected that the use of real-time PCR will increase in both diagnostics and in research, as a tool for target detection and quantification as well as for gene expression analysis.     

An improved method for high-quality RNA isolation from needles of adult maritime pine trees

When conventional RNA isolation methods optimized for pine seedlings are applied to needles of adult pine trees, poor-quality RNA results. Here we describe a modified procedure to isolate high-quality RNA from needles of 30-year-old maritime pines, exhibiting high levels of phenolics, polysaccharides, and RNases. Major changes are the inclusion of proteinase K in the extraction medium followed by incubation at 42°C. Integrity and purity were evaluated by using denaturing gel electrophoresis and spectrophotometry (A₂₆₀/A₂₃₀ and A₂₆₀/A₂₈₀). The total RNA could be successfully used for poly(A)⁺-RNA isolation and cDNA library construction.     

A new method of methyl methacrylate fixation in skull reconstruction

Fifty-two methyl methacrylate cranioplasties were performed in 47 patients over a 5-year period. The indications for the original craniectomy fell into four categories. Nine patients had a craniectomy for tumor, 32 for trauma, 5 for infection, and 1 for aneurysm. A new approach, which eliminates the need for suture or wire fixation and which facilitates intraoperative calvarial contouring, is presented. An excellent cosmetic result is the norm with this technique.     

A simple method for forehead unit reconstruction

The supratrochlear and supraorbital vessel-based island flap was used to avoid excessive scarring during reconstruction of the unilateral forehead defects after surgical excision of a malignant tumor. This method had a number of advantages: operation time was short, it was a one-stage procedure, the technique was relatively simple, a good match of the texture and color could be achieved, and scars were inconspicuous.     

A single-stage two-flap method of total ear reconstruction

A single-stage two-flap method of total ear reconstruction in congenital microtia is reported. This method was derived from the one-stage reconstruction described by Song and Song. Two flaps defined by vascular basis were elevated on the mastoid area: the superficial skin flap supplied mainly by subcutaneous pedicled arteriole perforators from the posterior auricular artery and the deeper axial-pattern fascial flap including the posterior auricular artery itself. The ear framework, exaggeratedly carved using autologous rib cartilage, could be inserted easily between the two flaps, simultaneously producing the auriculocephalic angle and the conchal wall. Intraoperative expansion of the skin flap and postoperative external ear molding also were performed to create aesthetically pleasing ears.     

A simple method for alar rim reconstruction

A one-stage repair procedure is described for the reconstruction of small to medium-sized full-thickness alar rim defects with a hinged medially based nasolabial island flap. The operation is performed under local anesthesia as an office procedure and is indicated particularly in older patients. The hinged flap provides both the inner and outer layers of the alar rim. The donor site is closed primarily with no need to mobilize a large skin flap. The procedure leaves no conspicuous scars on the face. This method was found to be simple and safe, providing excellent tissue viability and yielding good color and texture match.     

Induced fit in arginine kinase

Creatine kinase (CK) and arginine kinase (AK) are related enzymes that reversibly transfer a phosphoryl group between a guanidino compound and ADP. In the buffering of ATP energy levels, they are central to energy metabolism and have been paradigms of classical enzymology. Comparison of the open substrate-free structure of CK and the closed substrate-bound structure of AK reveals differences that are consistent with prior biophysical evidence of substrate-induced conformational changes. Large and small domains undergo a hinged 13 degrees rotation. Several loops become ordered and adopt different positions in the presence of substrate, including one (residues 309-319) that moves 15 A to fold over the substrates. The conformational changes appear to be necessary in aligning the two substrates for catalysis, in configuring the active site only when productive phosphoryl transfer is possible, and excluding water from the active site to avoid wasteful ATP hydrolysis.