Uncertainty in positioning ion chamber at reference depth for various water phantoms

Background

Uncertainty in the calibration of high-energy radiation sources is dependent on user and equipment type.

Numerical investigation of thermal response of laser-irradiated biological tissue phantoms embedded with gold nanoshells

The work presented in this paper focuses on numerically investigating the thermal response of gold nanoshells-embedded biological tissue phantoms with potential applications into photo-thermal therapy wherein the interest is in destroying the cancerous cells with minimum damage to the surrounding healthy cells. The tissue phantom has been irradiated with a pico-second laser. Radiative transfer equation (RTE) has been employed to model the light-tissue interaction using discrete ordinate method (DOM). For determining the temperature distribution inside the tissue phantom, the RTE has been solved in combination with a generalized non-Fourier heat conduction model namely the dual phase lag bio-heat transfer model. The numerical code comprising the coupled RTE-bio-heat transfer equation, developed as a part of the current work, has been benchmarked against the experimental as well as the numerical results available in the literature. It has been demonstrated that the temperature of the optical inhomogeneity inside the biological tissue phantom embedded with gold nanoshells is relatively higher than that of the baseline case (no nanoshells) for the same laser power and operation time. The study clearly underlines the impact of nanoshell concentration and its size on the thermal response of the biological tissue sample. The comparative study concerned with the size and concentration of nanoshells showed that 60 nm nanoshells with concentration of 5×10¹⁵ mm⁻³ result into the temperature levels that are optimum for the irreversible destruction of cancer infected cells in the context of photo-thermal therapy. To the best of the knowledge of the authors, the present study is one of the first attempts to quantify the influence of gold nanoshells on the temperature distributions inside the biological tissue phantoms upon laser irradiation using the dual phase lag heat conduction model.     

Nanoparticle-aided external beam radiotherapy leveraging the Čerenkov effect

This study investigates the feasibility of exploiting the Čerenkov radiation (CR) present during external beam radiotherapy (EBRT) for significant therapeutic gain, using titanium dioxide (titania) nanoparticles (NPs) delivered via newly designed radiotherapy biomaterials. Using Monte Carlo radiation transport simulations, we calculated the total CR yield inside a tumor volume during EBRT compared to that of the radionuclides. We also considered a novel approach for intratumoral titania delivery using radiotherapy biomaterials (e.g. fiducials) loaded with NPs. The intratumoral distribution/diffusion of titania released from the fiducials was calculated. To confirm the CR induced enhancement in EBRT experimentally, we used 6 MV radiation to irradiate human lung cancer cells with or without titania NPs and performed clonogenic assays. For a radiotherapy biomaterial loaded with 20 μg/g of 2-nm titania NPs, at least 1 μg/g could be delivered throughout a tumor sub-volume of 2-cm diameter after 14 days. This concentration level could inflict substantial damage to cancer cells during EBRT. The Monte Carlo results showed the CR yield by 6 MV radiation was higher than by the radionuclides of interest and hence greater damage might be obtained during EBRT. In vitro study showed significant enhancement with 6 MV radiation and titania NPs. These preliminary findings demonstrate a potential new approach that can be used to take advantage of the CR present during megavoltage EBRT to boost damage to cancer cells. The results provide significant impetus for further experimental studies towards the development of nanoparticle-aided EBRT powered by the Čerenkov effect.     

Shape complexity of space used by American black bears influenced by sex and intensity of use

Disentangling the complexities that influence animal space use poses substantial challenges based on decision trade-offs and constraints imposed on animals. Optimal decisions suggest that the spatial complexity of home-range shapes should be inversely related to energy conservation and fitness. Hence, the most beneficial shape should be the circle. We evaluated whether shape complexity (i.e., an index of circularity) of home ranges was influenced by two extrinsic (spatial heterogeneity, preferred habitat [i.e., deciduous forest]), and three intrinsic (sex, season [breeding, non-breeding], intensity of use) factors, with intensity of use indexed as contours containing core and peripheral areas. We estimated utilization distributions of 39 radiomarked adult American black bears (Ursus americanus), a habitat generalist, using fixed-kernel techniques and estimated 50% (core area) and 95% (peripheral area) contours. We fit a set of 47 models using linear modeling and ranked models using small-sample Akaike Information Criterion. Coefficients for the best model were the intrinsic factors intensity of use (reference category = core; 0.118; 95% CL = 0.064–0.173), sex (reference = female; 0.105; 95% CL = 0.043–0.167), and intercept (0.229; 95% CL = 0.186–0.272). Shape complexity was less for core areas than peripheral areas and less for females than males. Considering complex resource selection patterns within a fragmented landscape, both sexes appeared to use energy-maximizing strategies, although the increase in shape complexity for males may be an allometric relationship based on size dimorphism. Our approach supported the phenomenon of optimality as manifested through home-range shape complexity, but we suggest that assessment of this phenomenon for habitat specialists may yield different results, including the potential importance of intrinsic factors based on more restrictive limiting factors.     

CT dose reduction using Automatic Exposure Control and iterative reconstruction: A chest paediatric phantoms study

PurposeTo evaluate the impact of Automatic Exposure Control (AEC) on radiation dose and image quality in paediatric chest scans (MDCT), with or without iterative reconstruction (IR).MethodsThree anthropomorphic phantoms representing children aged one, five and 10-year-old were explored using AEC system (CARE Dose 4D) with five modulation strength options. For each phantom, six acquisitions were carried out: one with fixed mAs (without AEC) and five each with different modulation strength. Raw data were reconstructed with Filtered Back Projection (FBP) and with two distinct levels of IR using soft and strong kernels. Dose reduction and image quality indices (Noise, SNR, CNR) were measured in lung and soft tissues. Noise Power Spectrum (NPS) was evaluated with a Catphan 600 phantom.ResultsThe use of AEC produced a significant dose reduction (p < 0.01) for all anthropomorphic sizes employed. According to the modulation strength applied, dose delivered was reduced from 43% to 91%. This pattern led to significantly increased noise (p < 0.01) and reduced SNR and CNR (p < 0.01). However, IR was able to improve these indices. The use of AEC/IR preserved image quality indices with a lower dose delivered. Doses were reduced from 39% to 58% for the one-year-old phantom, from 46% to 63% for the five-year-old phantom, and from 58% to 74% for the 10-year-old phantom. In addition, AEC/IR changed the patterns of NPS curves in amplitude and in spatial frequency.ConclusionsIn chest paediatric MDCT, the use of AEC with IR allows one to obtain a significant dose reduction while maintaining constant image quality indices.     

Development of a high resolution voxelised head phantom for medical physics applications

Computational anthropomorphic phantoms have become an important investigation tool for medical imaging and dosimetry for radiotherapy and radiation protection. The development of computational phantoms with realistic anatomical features contribute significantly to the development of novel methods in medical physics. For many applications, it is desirable that such computational phantoms have a real-world physical counterpart in order to verify the obtained results.In this work, we report the development of a voxelised phantom, the HIGH_RES_HEAD, modelling a paediatric head based on the commercial phantom 715-HN (CIRS). HIGH_RES_HEAD is unique for its anatomical details and high spatial resolution (0.18 × 0.18 mm² pixel size). The development of such a phantom was required to investigate the performance of a new proton computed tomography (pCT) system, in terms of detector technology and image reconstruction algorithms.The HIGH_RES_HEAD was used in an ad-hoc Geant4 simulation modelling the pCT system. The simulation application was previously validated with respect to experimental results. When compared to a standard spatial resolution voxelised phantom of the same paediatric head, it was shown that in pCT reconstruction studies, the use of the HIGH_RES_HEAD translates into a reduction from 2% to 0.7% of the average relative stopping power difference between experimental and simulated results thus improving the overall quality of the head phantom simulation.The HIGH_RES_HEAD can also be used for other medical physics applications such as treatment planning studies.A second version of the voxelised phantom was created that contains a prototypic base of skull tumour and surrounding organs at risk.     

Effect of long-term exposure to mobile phone radiation on alpha-Int1 gene sequence of Candida albicans

Over the last decade, communication industries have witnessed a tremendous expansion, while, the biological effects of electromagnetic waves have not been fully elucidated. Current study aimed at evaluating the mutagenic effect of long-term exposure to 900-MHz radiation on alpha-Int1 gene sequences of Candida albicans. A standard 900 MHz radiation generator was used for radiation. 10 ml volumes from a stock suspension of C. albicans were transferred into 10 polystyrene tubes. Five tubes were exposed at 4 °C to a fixed magnitude of radiation with different time periods of 10, 70, 210, 350 and 490 h. The other 5 tubes were kept far enough from radiation. The samples underwent genomic DNA extraction. PCR amplification of alpha-Int1 gene sequence was done using one set of primers. PCR products were resolved using agarose gel electrophoresis and the nucleotide sequences were determined. All samples showed a clear electrophoretic band around 441 bp and further sequencing revealed the amplified DNA segments are related to alpha-Int1 gene of the yeast. No mutations in the gene were seen in radiation exposed samples. Long-term exposure of the yeast to mobile phone radiation under the above mentioned conditions had no mutagenic effect on alpha-Int1 gene sequence.     

Visualization of air and metal inhomogeneities in phantoms irradiated by carbon ion beams using prompt secondary ions

PurposeNon-invasive methods for monitoring of the therapeutic ion beam extension in the patient are desired in order to handle deteriorations of the dose distribution related to changes of the patient geometry. In carbon ion radiotherapy, secondary light ions represent one of potential sources of information about the dose distribution in the irradiated target. The capability to detect range-changing inhomogeneities inside of an otherwise homogeneous phantom, based on single track measurements, is addressed in this paper.MethodsAir and stainless steel inhomogeneities, with PMMA equivalent thickness of 10 mm and 4.8 mm respectively, were inserted into a PMMA-phantom at different positions in depth. Irradiations of the phantom with therapeutic carbon ion pencil beams were performed at the Heidelberg Ion Beam Therapy Center. Tracks of single secondary ions escaping the phantom under irradiation were detected with a pixelized semiconductor detector Timepix. The statistical relevance of the found differences between the track distributions with and without inhomogeneities was evaluated.ResultsMeasured shifts of the distal edge and changes in the fragmentation probability make the presence of inhomogeneities inserted into the traversed medium detectable for both, 10 mm air cavities and 1 mm thick stainless steel. Moreover, the method was shown to be sensitive also on their position in the observed body, even when localized behind the Bragg-peak.ConclusionsThe presented results demonstrate experimentally, that the method using distributions of single secondary ion tracks is sensitive to the changes of homogeneity of the traversed material for the studied geometries of the target.     

Predicted ionisation in mitochondria and observed acute changes in the mitochondrial transcriptome after gamma irradiation: A Monte Carlo simulation and quantitative PCR study

It is a widely accepted that the cell nucleus is the primary site of radiation damage while extra-nuclear radiation effects are not yet systematically included into models of radiation damage.We performed Monte Carlo simulations assuming a spherical cell (diameter 11.5 μm) modelled after JURKAT cells with the inclusion of realistic elemental composition data based on published literature. The cell model consists of cytoplasm (density 1 g/cm³), nucleus (diameter 8.5 μm; 40% of cell volume) as well as cylindrical mitochondria (diameter 1 μm; volume 0.5 μm³) of three different densities (1, 2 and 10 g/cm³) and total mitochondrial volume relative to the cell volume (10, 20, 30%). Our simulation predicts that if mitochondria take up more than 20% of a cell's volume, ionisation events will be the preferentially located in mitochondria rather than in the cell nucleus.Using quantitative polymerase chain reaction, we substantiate in JURKAT cells that human mitochondria respond to gamma radiation with early (within 30 min) differential changes in the expression levels of 18 mitochondrially encoded genes, whereby the number of regulated genes varies in a dose-dependent but non-linear pattern (10 Gy: 1 gene; 50 Gy: 5 genes; 100 Gy: 12 genes).The simulation data as well as the experimental observations suggest that current models of acute radiation effects, which largely focus on nuclear effects, might benefit from more systematic considerations of the early mitochondrial responses and how these may subsequently determine cell response to ionising radiation.     

Application of the phase-space distribution approach of Monte Carlo for radiation contamination dose estimation from the (n,γ), (γ,n) nuclear reactions and linac leakage photons in the megavoltage radiotherapy facility

AimThe aim of this study was to characterize the radiation contamination inside and outside the megavoltage radiotherapy room.BackgroundRadiation contamination components in the 18 MV linac room are the secondary neutron, prompt gamma ray, electron and linac leakage radiation.Materials and MethodsAn 18 MV linac modeled in a typical bunker employing the MCNPX code of Monte Carlo. For fast calculation, phase-space distribution (PSD) file modeling was applied and the calculations were conducted for the radiation contamination components dose and spectra at 6 locations inside and outside the bunker.ResultsThe results showed that the difference of measured and calculated percent depth-dose (PDD) and photo beam-profile (PBP) datasets were lower than acceptable values. At isocenter, the obtained photon dose and neutron fluence were 2.4 × 10⁻¹⁴ Gy/initial e° and 2.22 × 10^-8 n°/cm², respectively. Then, neutron apparent source strength (Q_N) value was found as 1.34 × 10¹² n°/Gy X at isocenter and the model verified to photon and neutron calculations. A surface at 2 cm below the flattening filter was modeled as phase-space (PS) file for PDD and PBP calculations. Then by use of a spherical cell in the center of the linac target as a PS surface, contaminant radiations dose, fluence and spectra were estimated at 6 locations in a considerably short time, using the registered history of all particles and photons in the 13GB PSD file as primary source in the second step.ConclusionDesigning the PSD file in MC modeling helps user to solve the problems with complex geometry and physics precisely in a shorter run-time.     

Cancer risk estimation in Digital Breast Tomosynthesis using GEANT4 Monte Carlo simulations and voxel phantoms

The aim of this work was to estimate the risk of radiation induced cancer following the Portuguese breast screening recommendations for Digital Mammography (DM) when applied to Digital Breast Tomosynthesis (DBT) and to evaluate how the risk to induce cancer could influence the energy used in breast diagnostic exams. The organ doses were calculated by Monte Carlo simulations using a female voxel phantom and considering the acquisition of 25 projection images. Single organ cancer incidence risks were calculated in order to assess the total effective radiation induced cancer risk. The screening strategy techniques considered were: DBT in Cranio-Caudal (CC) view and two-view DM (CC and Mediolateral Oblique (MLO)).The risk of cancer incidence following the Portuguese screening guidelines (screening every two years in the age range of 50–80 years) was calculated by assuming a single CC DBT acquisition view as standalone screening strategy and compared with two-view DM. The difference in the total effective risk between DBT and DM is quite low. Nevertheless in DBT an increase of risk for the lung is observed with respect to DM. The lung is also the organ that is mainly affected when non-optimal beam energy (in terms of image quality and absorbed dose) is used instead of an optimal one. The use of non-optimal energies could increase the risk of lung cancer incidence by a factor of about 2.     

The consideration of biological effectiveness of low energy protons using biophysical modeling of the effects induced by exposure of V79 cells

We have applied Monte Carlo track structure simulations to estimate relative biological effectiveness (RBE) of low-energy protons using biophysical modelling of radiation effects induced by exposure of V79 cells growing in mono-layer. The microscopic energy deposition in cell nucleus and sub-nucleus volumes was investigated in order to understand the reasons of enhanced biological effectiveness near Bragg peak. Theoretical estimations of RBE based on frequency/dose average lineal energy and calculated yields of initial DNA breaks were collated with experimental RBE_M data. It was found: (1) dose average lineal energy for whole cell nucleus as a function of proton energy shows a distinct peak at 550 keV; (2) the peak values for subnucleus volumes are large compared with the whole cell nucleus; (3) the yield of complex DNA breaks correlates with experimental RBE_M data.     

Incident air kerma to absorbed organ dose conversion factors for breast and lung in PA thorax radiography: The effect of patient thickness and radiation quality

PurposeConverting the measurable quantities to patient organ doses in projection radiography is usually based on a standard-sized patient model and a specific radiation quality, which are likely to differ from the real situation. Large inaccuracies can therefore be obtained in organ doses, because organ doses are dependent on the exposure parameters, exposure geometry and patient anatomy. In this study, the effect of radiation quality and patient thickness on the organ dose conversion factors were determined.MethodsIn this study, the posterior–anterior projection radiograph of the thorax was selected in order to determine the effect of radiation quality (tube voltages of 70–130 kV and total filtrations of 3 mmAl to 4 mmAl + 0.2 mmCu) and patient thickness (anterior–posterior thicknesses of 19.4–30.8 cm) on the breast and lung dose conversion factors. For this purpose, Monte Carlo simulation programs ImpactMC and PCXMC were used with computed tomography examination data of adult male and female patients and mathematical hermaphrodite phantoms, respectively.ResultsCompared to the reference beam quality and patient thickness, the relative variation range in organ dose conversion factors was up to 74% for different radiation qualities and 122% for different patient thicknesses.ConclusionsConversion factors should only be used with comprehensive understanding of the exposure conditions, considering the exposure parameters, exposure geometry and patient anatomy they are valid for. This study demonstrates that patient thickness-specific and radiation quality-specific conversion factors are needed in projection radiography.     

Drag force mechanical power during an actual propulsion cycle on a manual wheelchair

The object of this study was to compute the mechanical power of the resultant braking force during an actual propulsion cycle with a manual wheelchair on the field. The resultant braking force was calculated from a mechanical model taking into account the rolling resistances of the front and rear wheels. Both the resultant braking force and the wheelchair velocity were not constant during the propulsion cycle and varied according to the subject's fore-and-aft and vertical movements in the wheelchair. These variations had logical repercussions on the braking force mechanical power, which ranged from 20.6 to 34.5 W (mean = 29.6 W) during the propulsion cycle. The mechanical power was also calculated from the conditions of a classical drag test, by the product of the cycle mean velocity and a constant braking force corresponding to a 60% rear wheels distribution of the subject-and-wheelchair's weight. This second mechanical power (32.4 W) was 10% higher than the average of the instantaneous power. Beyond the need of a clear definition of the two phases of the propulsion cycle, this study showed that the assumption on wheelchair locomotion usually admitted on laboratory ergometers cannot be applied in field studies, and that the kinetic energy variations during the cycle propulsive phase should be considered for evaluating the subject's mechanical work and power.     

Impaired NHEJ function in multiple myeloma

Multiple myeloma (MM) is characterized by multiple chromosomal aberrations. To assess the contribution of DNA repair to this phenotype, ionizing radiation was used to induce DNA double strand breaks in three MM cell lines. Clonogenic survival assays showed U266 (SF4 = 15.3 + 6.4%) and RPMI 8226 (SF4 = 12.6.0 + 1.7%) were radiation sensitive while OPM2 was resistant (SF4 = 78.9 + 4.1%). Addition of the DNA-PK inhibitor NU7026 showed the expected suppression in radiation survival in OPM2 but increased survival in both radiation sensitive cell lines. To examine non-homologous end joining (NHEJ) repair in these lines, the ability of protein extracts to support in vitro DNA repair was measured. Among the three MM cell lines analyzed, RPMI 8226 demonstrated impaired blunt ended DNA ligation using a ligation-mediated PCR technique. In a bacterial based functional assay to rejoin a DNA break within the β-galactosidase gene, RPMI 8226 demonstrated a 4-fold reduction in rejoining fidelity compared to U266, with OPM2 showing an intermediate capacity. Ionizing radiation induced a robust γ-H2AX response in OPM2 but only a modest increase in each radiation sensitive cell line perhaps related to the high level of γ-H2AX in freshly plated cells. Examination of γ-H2AX foci in RPMI 8226 cells confirmed data from Western blots where a significant number of foci were present in freshly plated untreated cells which diminished over 24 h of culture. Based on the clonogenic survival and functional repair assays, all three cell lines exhibited corrupt NHEJ repair. We conclude that suppression of aberrant NHEJ function using the DNA-PK inhibitor NU7026 may facilitate access of DNA ends to an intact homologous recombination repair pathway, paradoxically increasing survival after irradiation. These data provide insight into the deregulation of DNA repair at the site of DNA breaks in MM that may underpin the characteristic genomic instability of this disease.     

Quantification of nortriptyline in plasma by HPLC and fluorescence detection

A simple, sensitive and specific high-performance liquid chromatography method has been developed for the determination of nortriptyline (NT) in plasma samples. The assay involved derivatization with 9H-fluoren-9-ylmethyl chloroformate (Fmoc-Cl) and isocratic reversed-phase (C₁₈) chromatography with fluorescence detection. The developed method required only 100 μl of plasma sample, deproteinized and derivatized in one step. Calibration curves were lineal over the concentration range of 5–5000 ng/ml. The derivatization reaction was performed at room temperature in 20 min and the obtained NT derivative was stable for at least 48 h at room temperature. The within-day and between-day relative standard deviation was below 8%. The limit of detection (LOD) was 2 ng/ml, and the lower limit of quantification (LLOQ) was established at 10 ng/ml. The method was applied on plasma collected from rats, at different time intervals, after intravenous administration of 0.5 mg of NT.     

Dosimetric impact on changes in target volumes during intensity-modulated radiotherapy for nasopharyngeal carcinoma

Background and purposeTo assess anatomic changes during intensity modulated radiotherapy (IMRT) for nasopharyngeal carcinoma (NPC) and to determine its dosimetric impact.Patients and methodsTwenty patients treated with IMRT for NPC were enrolled in this study. A second CT was performed at 38 Gy. Manual contouring of the macroscopic tumor volumes (GTV) and the planning target volumes (PTV) were done on the second CT. We recorded the volumes of the different structures, D98 %, the conformity, and the homogeneity indexes for each PTV. Volume percent changes were calculated.ResultsWe observed a significant reduction in tumor volumes (58.56 % for the GTV N and 29.52 % for the GTV T). It was accompanied by a significant decrease in the D98 % for the 3 PTV (1.4 Gy for PTV H, p = 0.007; 0.3 Gy for PTV I, p = 0.03 and 1.15 Gy for PTV L, p = 0 0.0066). In addition, we observed a significant reduction in the conformity index in the order of 0.02 (p = 0.001) and 0.01 (p = 0.007) for PTV H and PTV I, respectively. The conformity variation was not significant for PTV L. Moreover, results showed a significant increase of the homogeneity index for PTV H (+ 0.03, p = 0.04) and PTV L (+ 0.04, p = 0.01).ConclusionTumor volume reduction during the IMRT of NPC was accompanied by deterioration of the dosimetric coverage for the different target volumes. It is essential that a careful adaptation of the treatment plan be considered during therapy for selected patients.     

Acute effects of incremental inspiratory loads on compartmental chest wall volume and predominant activity frequency of inspiratory muscle

AimThis research aims to analyze the acute effect of incremental inspiratory loads on respiratory pattern and on the predominant activity frequency of inspiratory muscle, taking into account differences in gender responses. Optoelectronic Plethysmography was performed during loads in 39 healthy subjects (20 women), placing 89 markers on the thoracic-abdominal wall to obtain total and regional volumes. Surface electromyography (SEMG) was taken simultaneously on the Sternocleidomastoid and Diaphragm muscles, to calculate the predominant muscle activity frequency through wavelet analysis. Inspiratory loads were performed using Threshold® with 2 min of breathing at different levels, ranging from a load of 10 cmH₂O plus 5 cmH₂O to 40 cmH₂O or fatigue.ResultsInspiratory Time increased during loads. Total and compartmental volumes increased with different regions, changing at different loads. These changes in volume occur earlier in women (20 cmH₂O) than in men (30 cmH₂O). The predominant activity frequency of Sternocleidmastoid muscle decreased at 30 cmH₂O, while Diaphragm activity decreased at 40 cmH₂O.ConclusionThe acute effects of incremental inspiratory loads are increases of total and regional volumes and inspiratory time. As for muscle activity, the predominant activity frequency declined in Sternocleidomastoid and Diaphragm muscles, but at different loads. Such respiratory and SEMG patterns and gender differences should be considered when clinical interventions are performed.     

Polymer gel dosimeters for pretreatment radiotherapy verification using the three-dimensional gamma evaluation and pass rate maps

Polymer gel dosimeters (PGDs) have been widely studied for use in the pretreatment verification of clinical radiation therapy. However, the readability of PGDs in three-dimensional (3D) dosimetry remain unclear. In this study, the pretreatment verifications of clinical radiation therapy were performed using an N-isopropyl-acrylamide (NIPAM) PGD, and the results were used to evaluate the performance of the NIPAM PGD on 3D dose measurement. A gel phantom was used to measure the dose distribution of a clinical case of intensity-modulated radiation therapy. Magnetic resonance imaging scans were performed for dose readouts. The measured dose volumes were compared with the planned dose volume. The relative volume histograms showed that relative volumes with a negative percent dose difference decreased as time elapsed. Furthermore, the histograms revealed few changes after 24 h postirradiation. For the 3%/3 mm and 2%/2 mm criteria, the pass rates of the 12- and 24-h dose volumes were higher than 95%, respectively. This study thus concludes that the pass rate map can be used to evaluate the dose-temporal readability of PGDs and that the NIPAM PGD can be used for clinical pretreatment verifications.     

Ontogeny of the Spitzenkörper in germlings of Neurospora crassa

The Spitzenkörper (Spk) is a highly dynamic and pleomorphic complex located at the hyphal apex of filamentous fungi. Most studies revealing the structure and behavior of the Spk have been conducted on mature vegetative hyphae of filamentous fungi, including both main leading hyphae and branches. However, these reports do not address whether the observations can be extended to germ tubes. By enhanced phase-contrast video-microscopy and laser scanning confocal microscopy we have analyzed the intracellular changes prior to the appearance of a Spk in germlings of Neurospora crassa. Observations began at the early stages of spore germination and were carried out until a conspicuous Spk could be observed at the apex of germ tubes. Before a Spk could be observed, young germ tubes (<150 μm) displayed a uniform distribution of organelles such as nuclei, mitochondria, and cytoplasmic granules along the length of the cells. Once the germlings started reaching lengths of more than approximately 150 μm, visible organelles experienced a displacement towards the subapical region of the cell and a small exclusion zone free of organelles (0.6 ± 0.3 μm) formed at the apex. The position of this exclusion zone within the apex seemed to determine the germling growth direction, which was highly erratic. Few minutes after it first appeared, upon growth of the germling, the exclusion zone started to become occupied by an accumulation of material that gradually concentrated into a light gray body that we describe as an immature Spk. During this phase the presence of a Spk in the apical dome was not constant. Approximately 30 min later, the immature Spk became more robust and gradually acquired its typical phase-dark appearance, while the growth direction of the germ tube became less wavering. The formation of a mature phase-dark Spk coincided with the stabilization of the growth direction of the germling, therefore suggesting that it is at this stage when the transition from germling to vegetative hypha occurs.