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.     

Review of strategies for MRI based reconstruction of endocavitary and interstitial applicators in brachytherapy of cervical cancer

Brachytherapy plays an essential role in the curative intent management of locally advanced cervical cancer. The introduction of the magnetic resonance (MR) as a preferred image modality and the development of new type of applicators with interstitial components have further improved its benefits.The aim of this work is to review the current status of one important aspect in the cervix cancer brachytherapy procedure, namely catheter reconstruction.MR compatible intracavitary and interstitial applicators are described. Considerations about the use of MR imaging (MRI) regarding appropriate strategies for applicator reconstruction, technical requirements, MR sequences, patient preparation and applicator commissioning are included.It is recommendable to perform the reconstruction process in the same image study employed by the physician for contouring, that is, T2 weighted (T2W) sequences. Nevertheless, a clear identification of the source path inside the catheters and the applicators is a challenge when using exclusively T2W sequences. For the intracavitary component of the implant, sometimes the catheters may be filled with some substance that produces a high intensity signal on MRI. However, this strategy is not feasible for plastic tubes or titanium needles, which, moreover, induce magnetic susceptibility artifacts. In these situations, the use of applicator libraries available in the treatment planning system (TPS) is useful, since they not only include accurate geometrical models of the intracavitary applicators, but also recent developments have made possible the implementation of the interstitial component. Another strategy to improve the reconstruction process is based on the incorporation of MR markers, such as small pellets, to be used as anchor points.Many institutions employ computed tomography (CT) as a supporting image modality. The registration of CT and MR image sets should be carefully performed, and its uncertainty previously assessed. Besides, an important research work is being carried out regarding the use of ultrasound and electromagnetic tracking technologies.     

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.     

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.     

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.     

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.     

Multi-parametric neuroimaging evaluation of cerebrotendinous xanthomatosis and its correlation with neuropsychological presentations

Background

Cerebrotendinous xanthomatosis (CTX) is a rare genetic disorder. Recent studies show that brain damage in CTX patients extends beyond the abnormalities observed on conventional magnetic resonance imaging (MRI). We studied the MRI and ^{99 m}Tc-ethyl cysteinate dimer single photon emission computed tomography (SPECT) findings of CTX patients and made a correlation with the neuropsychological presentations.

Methods

Diffusion tensor imaging (DTI) and 3D T1-weighted images of five CTX patients were compared with 15 age-matched controls. Voxel-based morphometry (VBM) was use to delineate gray matter (GM) and white matter (WM) volume loss. Fractional anisotropy (FA), mean diffusivity (MD), and eigenvalues derived from DTI were used to detect WM changes and correlate with neuropsychological results. SPECT functional studies were used to correlate with GM changes.

Results

Cognitive results showed that aside from moderate mental retardation, the patient group performed worse in all cognitive domains. Despite the extensive GM atrophy pattern, the cerebellum, peri-Sylvian regions and parietal-occipital regions were correlated with SPECT results. WM atrophy located in the peri-dentate and left cerebral peduncle areas corresponded with changes in diffusion measures, while axial and radial diffusivity suggested both demyelinating and axonal changes. Changes in FA and MD were preceded by VBM in the corpus callosum and corona radiata. Cognitive results correlated with FA changes.

Conclusion

In CTX, GM atrophy affected the perfusion patterns. Changes in WM included atrophy, and axonal changes with demyelination. Disconnection of major fiber tracts among different cortical regions may contribute to cognitive impairment.

     

剪叶损伤与昆虫取食对兴安落叶松(Larix gmelinii)针叶中缩合单宁诱导作用的差异

单宁是植物内与抗虫性相关的重要次生代谢物质.为了研究昆虫取食及剪叶诱导对兴安落叶松(Larix gmelinii)针叶内缩合单宁含量的影响,用剪叶法和落叶松毛虫(Dendrolimus superans)幼虫取食处理5年生兴安落叶松苗,以香草醛-盐酸法测定其剩余健康针叶内缩合单宁含量的变化.结果表明:(1)处理后1～10d,剪叶4枝75%,8枝50%、75%,12枝25%、50%、75%及虫伤4枝75%的兴安落叶松健康针叶内的缩合单宁含量均明显高于对照(P<0.01),15d时,剪叶8枝75%,12枝50%的兴安落叶松针叶内的缩合单宁含量仍高于对照(P<0.05),说明剪叶与虫伤处理均能诱导缩合单宁含量增加.(2)剪叶4枝75%,8枝50%,12枝25%、75%的处理之间,诱导的缩合单宁含量差异不显著(P>0.05).剪叶8枝75%,12枝50%诱导缩合单宁含量增加幅度比其它剪叶处理大,且诱导持续时间长.说明剪叶程度虽能影响缩合单宁含量,但二者并不呈线性关系.(3)剪叶4枝75%在5d时诱导缩合单宁含量最高,虫伤4枝75%在10d时诱导缩合单宁含量最高,说明剪叶诱导的缩合单宁含量高峰早于虫伤处理.但处理后1～10d,剪叶及虫伤4枝75%的落叶松针叶内缩合单宁含量均明显高于对照(P<0.01或P<0.05),之后与对照差异不显著(P>0.05),说明在受到诱导处理后,兴安落叶松产生应激反应,使其针叶内缩合单宁含量在一定时间内先增加,后逐渐恢复到正常水平.由上可见,可以采取适当的损伤处理取得与昆虫取食相似的兴安落叶松抗性反应.     

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 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.