Matching and reconstruction of brachytherapy seeds using the Hungarian algorithm (MARSHAL)

Intraoperative dosimetric quality assurance in prostate brachytherapy critically depends on discerning the three-dimensional (3D) locations of implanted seeds. The ability to reconstruct the implanted seeds intraoperatively will allow us to make immediate provisions for dosimetric deviations from the optimal implant plan. A method for seed reconstruction from segmented C-arm fluoroscopy images is proposed. The 3D coordinates of the implanted seeds can be calculated upon resolving the correspondence of seeds in multiple x-ray images. We formalize seed-matching as a combinatorial optimization problem, which has salient features: (a) extensively studied solutions by the computer science community; (b) proof for the nonexistence of any polynomial time exact algorithm; and (c) a practical pseudo-polynomial algorithm that mostly runs in O(N3) time using any number of images. We prove that two images are insufficient to correctly match the seeds, while a third image renders the matching problem to be of nonpolynomial complexity. We utilize the special structure of the problem and propose a pseudopolynomial time algorithm. Using three presegmented images, matching and reconstruction of brachytherapy seeds using the Hungarian algorithm achieved complete matching in simulation experiments; and 98.5% in phantom experiments. 3D reconstruction error for correctly matched seeds has a mean of 0.63 mm, and 0.9 mm for incorrectly matched seeds. The maximum seed reconstruction error in each implant was typically around 1.32 mm. Both on synthetic data and in phantom experiments, matching rate and reconstruction error achieved using presegmented images was found to be sufficient for prostate brachytherapy. The algorithm is extendable to deal with arbitrary number of images without any loss in speed or accuracy. The algorithm is sufficiently generic to provide a practical solution to any correspondence problem, across different imaging modalities and features.     

Umbilical cord prostaglandins in term and preterm parturition

Objective: Prostaglandins (PGs) are considered the universal mediators of parturition. Amniotic fluid PGE₂ and PGF_2α concentrations increase before the onset of spontaneous labor at term, as well as during labor. This study was conducted to determine if the concentrations of umbilical cord PGE₂ and PGF₂α change with advancing gestational age, spontaneous labor at term, and preterm labor (with and without funisitis).

Methods: Umbilical cord (UC) tissue samples were obtained from women (N?=?158) with singleton pregnancies in the following groups: (1) term deliveries without labor (TNL; n?=?20); (2) term deliveries with labor (TIL; n?=?20); (3) spontaneous preterm deliveries (sPTD) with (n?=?20) and without acute funisitis (n?=?20); and (4) preeclampsia without labor (n?=?78). The concentrations of PGs were determined in different locations of the UC. PGE₂ and PGF_2α were measured by specific immunoassays. Non-parametric statistics were used for analysis.

Results: (1) In spontaneous preterm deliveries, the median UC PGE₂ concentration was higher in cases with funisitis than in those without funisitis (233.7?pg/µg versus 87.4?pg/µg of total protein, p?=?0.001); (2) the median UC PGE₂ concentration in sPTD with funisitis was also higher than that obtained from samples who had undergone labor at term (233.7?pg/µg versus 116.1?pg/µg of total protein, p?=?0.03); (3) the UC PGE₂ and PGF_2α concentration increased as a function of advancing gestational age before 36 weeks (PGE₂: ρ?=?0.59, p?<?0.001; PGF_2α: ρ?=?0.39, p?=?0.01), but not after 36 weeks (PGE₂: ρ?=??0.1, p?=?0.5; PGF_2α: ρ?=??0.2, p?=?0.2); (4) the median UC concentrations of PGE₂ and PGF_2α at term was similar in samples obtained from women with and without labor (PGE₂: TNL 133.7?pg/µg versus TIL 116.1?pg/µg of total protein, p?=?0.9; PGF_2α: TNL 8.4?pg/µg versus TIL 8.1?pg/µg of total protein, p?=?0.7); and (5) there was no correlation between UC PG concentration and gestational age at term pregnancy (PGE₂: ρ?=?0.01, p?=?0.9; PGF_2α: ρ?=?0.07, p?=?0.7).

Conclusions: (1) PGE₂ concentrations in the UC are higher in the presence of acute funisitis than in the absence of this lesion; (2) spontaneous labor at term was not associated with a change in the UC concentration of PGE₂ and PGF_2α; and (3) the UC concentrations of PGE₂ and PGF_2α increased as a function of gestational age. We propose that UC PGs act as inflammatory mediators generated in the context of fetal systemic inflammation.     

Overnight changes of chemoreflex control in obstructive sleep apnoea patients

We hypothesized that the numerous episodes of hypoxia, hypercapnia and arousal experienced by obstructive sleep apnoea (OSA) patients induce overnight changes in respiratory chemoreflexes. A modification of the Read rebreathing technique assessed chemoreflex characteristics in the evening and the morning of patients undergoing diagnostic assessment for OSA in a clinical sleep laboratory. Two groups were studied: those with apnoea-hypopnoea indices (AHI) greater than 30 composed the OSA group (n = 12), and those with AHI indices less than 10 composed the non-OSA group (n = 12). There was a significant (approximately 30%) overnight increase in chemoreflex sensitivities, without changes in thresholds, in the OSA group. In the non-OSA group there was a significant overnight reduction in chemoreflex thresholds (approximately 5%), without changes in sensitivities. We suggest that these changes affect the stability of the chemoreflex control system in opposite ways as the night proceeds: destabilizing breathing for patients in the OSA group, and stabilising breathing for patients in the non-OSA group.     

Essential differences in ligand presentation and T cell epitope recognition among HLA molecules of the HLA-B44 supertype

Human leukocyte antigens (HLA) have long been grouped into supertypes to facilitate peptide-based immunotherapy. Analysis of several hundreds of peptides presented by all nine antigens of the HLA-B44 supertype (HLA-B*18, B*37, B*40, B*41, B*44, B*45, B*47, B*49 and B*50) revealed unique peptide motifs for each of them. Taking all supertype members into consideration only 25 out of 670 natural ligands were found on more than one HLA molecule. Further direct comparisons by two mass spectrometric methods--isotope labeling as well as a label-free approach--consistently demonstrated only minute overlaps of below 3% between the ligandomes of different HLA antigens. In addition, T cell reactions of healthy donors against immunodominant HLA-B*44 and HLA-B*40 epitopes from EBV lacked promiscuous T-cell recognition within the HLA-B44 supertype. Taken together, these results challenge the common paradigm of broadly presented epitopes within this supertype.     

Biotransport Phenomena in Freezing Mammalian Oocytes

Water transport across the cell plasma membrane and intracellular ice formation (IIF)—the two biophysical events that may cause cell injury during cryopreservation—were studied by cryomicroscopy and modeling using mammalian (Peromyscus) oocytes. Unusually high activation energy for water transport across the cell plasma membrane was identified indicating that the water transport process is unusually sensitive to temperature (and cooling rate). Although literally all studies on IIF were conducted using protocols with ice-seeding (seeding extracellular ice usually at ≥−7 °C), it is not used for cell cryopreservation by vitrification that is becoming increasingly popular today. In this article, we show that ice-seeding has a significant impact on IIF. With ice-seeding and cooling at 60 °C/min, IIF was observed to occur over a wide range from approximately −8 to −48 °C with a clear change of the ice nucleation mechanism (from surface- to volume-catalyzed nucleation) at approximately −43 °C. On the contrary, without ice-seeding, IIF occurred over a much narrower range from approximately −19 to −27 °C without a noticeable change of the nucleation mechanism. Moreover, the kinetics of IIF without ice-seeding was found to be strongly temperature (and cooling rate) dependent. These findings indicate the importance of quantifying the IIF kinetics in the absence of ice-seeding during cooling for development of optimal vitrification protocols of cell cryopreservation.     

Adventitia-derived hydrogen peroxide impairs relaxation of the rat carotid artery via smooth muscle cell p38 mitogen-activated protein kinase

The role of adventitia-derived reactive oxygen species (ROS) in vascular disease and impaired vascular relaxation is not clear. Based on robust adventitial ROS generation and effects on MAPK involvement in vascular dysfunction, we hypothesized that adventitia-derived ROS hydrogen peroxide (H(2)O(2)) impairs vascular relaxation through activation of medial smooth muscle p38 MAPK. By using a novel in vivo model, the adventitial surface of rat carotid arteries was bathed in situ for 90 min with vehicle, angiotensin II (AngII; 500 nM), AngII+H(2)O(2)-scavenger catalase (3,000 U/ml), AngII+p38 MAPK inhibitor SB203580 (10 μM), or AngII+superoxide dismutase (SOD; 150 U/ml). After these in vivo treatments, ex vivo tone measurements on isolated vessels revealed that periadventitial application of AngII impaired both acetylcholine-induced (endothelium-dependent) and sodium nitroprusside-induced (endothelium-independent) relaxations. In vivo coincubation with catalase or SB203580 significantly improved, but SOD exacerbated AngII-induced impairment of in vitro endothelium-dependent and -independent vascular relaxations. Western blots of vascular media, separated from the adventitia, demonstrated increased medial p38 MAPK activation and decreased medial phosphatase SHP-2 activity in AngII-treated vessels. These effects were reversed by in vivo periadventitial addition of catalase. These findings provide the first evidence that adventitia-derived H(2)O(2) participates in vascular dysfunction through p38 MAPK activation and SHP-2 inhibition.     

A novel temporal‐predominant neuro‐astroglial tauopathy associated with TMEM106B gene polymorphism in FTLD/ALS‐TDP

Polymorphisms in TMEM106B, a gene on chromosome 7p21.3 involved in lysosomal trafficking, correlates to worse neuropathological, and clinical outcomes in frontotemporal lobar degeneration (FTLD) and amyotrophic lateral sclerosis (ALS) with TDP‐43 inclusions. In a small cohort of C9orf72 expansion carriers, we previously found an atypical, neuroglial tauopathy in cases harboring a TMEM106B rs1990622 A/A genotype. To test whether TMEM106B genotype affects the risk of developing atypical tauopathy under a recessive genotype model (presence versus absence of two major alleles: A/A vs. A/G and G/G). We characterized the atypical tauopathy neuropathologically and determined its frequency by TMEM106B rs1990622 genotypes in 90 postmortem cases with a primary diagnosis of FTLD/ALS‐TDP [mean age at death 65.5 years (±8.1), 40% female]. We investigated the effect of this new atypical tauopathy on demographics and clinical and neuropsychological metrics. We also genotyped TMEM106B in an independent series with phenotypically similar cases. Sixteen cases (16/90, 17.7 %) showed the temporal‐predominant neuro‐astroglial tauopathy, and 93.7% of them carried an A/A genotype (vs. ~35% in a population cohort). The odds ratio of FTLD/ALS‐TDP individuals with the A/A genotype showing neuro‐astroglial tauopathy was 13.9. Individuals with this tauopathy were older at onset (p = 0.01). The validation cohort had a similarly high proportion of rs1990622 A/A genotype. TDP‐43 and tau changes co‐occur in a subset of neurons. Our data add to the growing body of evidence that TMEM106B polymorphisms may modulate neurodegeneration. A distinctive medial temporal predominant, 4‐repeat, neuro‐astroglial tauopathy strongly correlates to TMEM106B A/A genotype in FTLD/ALS‐TDP cases.     

Increased thymic B cells but maintenance of thymic structure, T cell differentiation and negative selection in lymphotoxin-alpha and TNF gene-targeted mice

TNF, lymphotoxin (LT) and their receptors are expressed constitutively in the thymus. It remains unclear whether these cytokines play a role in normal thymic structure or function. We have investigated thymocyte differentiation, selection and thymic organogenesis in gene targeted mice lacking LTalpha, TNF, or both (TNF/LTalpha-/-). The thymus was normal in TNF/LTalpha-/- mice with regard to cell yields and stromal architecture. Detailed analysis of alphabeta and gammadelta T cell-lineage thymocyte subsets revealed no abnormalities, implying that neither TNF nor LT play an essential role in T cell differentiation or positive selection. The number and distribution of thymic CD11c+ dendritic cells was also normal in the absence of both TNF and LTalpha. A three-fold increase in B cell numbers was observed consistently in the TNF/LTalpha-/- thymus. This phenotype was due entirely to the LTalpha deficiency and associated with changes in the hemopoietic compartment, rather than the thymic stromal compartment of LTalpha-/- mice. Finally, specific Vbeta8+ T cell deletion within the thymus following intrathymic injection of staphylococcal enterotoxin B (SEB) was TNF/LT independent. Thus, despite the presence of these cytokines and their receptors in the normal thymus, there appears no essential role for either TNF or LT in development of organ structure or for those processes associated with T cell repertoire selection.     

Healthcare worker competencies for disaster training

Background  

Although training and education have long been accepted as integral to disaster preparedness, many currently taught practices are neither evidence-based nor standardized. The need for effective evidence-based disaster training of healthcare staff at all levels, including the development of standards and guidelines for training in the multi-disciplinary health response to major events, has been designated by the disaster response community as a high priority. We describe the application of systematic evidence-based consensus building methods to derive educational competencies and objectives in criteria-based preparedness and response relevant to all hospital healthcare workers.