Apparent Fusion Research Articles

Differentiation of workers into soldiers is associated with a size reduction of higher-order brain centers in the neotropical termite Procornitermes araujoi

Comparing the size of functionally distinct brain regions across individuals with remarkable differences in sensory processing and cognitive demands provides important insights into the selective forces shaping animal nervous systems. We took advantage of the complex system of worker-to-soldier differentiation in the termitid Procornitermes araujoi, to investigate how a profound modification of body morphology followed by an irreversible shift in task performance are translated in terms of brain structure and size. This behavioural shift is characterised by a reduction of the once wide and complex behavioural repertoire of workers to one exclusively dedicated to nest defence (soldiers). In accordance with soldier’s reduced cognitive and sensory demands, we show here that differentiation of workers into soldiers is associated with a size reduction of the mushroom body (MB) compartments, higher-order brain regions responsible for multimodal processing and integration of sensory information, as well as learning, memory, and decision-making. Moreover, in soldiers, we found an apparent fusion of the medial and lateral MB calyces likely associated with its volume reduction. These results illustrate a functional neuroplasticity of the MB associated with division of labour, supporting the link between MB size and behavioural flexibility in social insect workers.

Fusion-fission analysis of C12+Cm248 and O16+Pu244 nuclear reactions across the Coulomb barrier

By using the symmetric-asymmetric Gaussian barrier distribution (SAGBD) model, Wong formula, and coupled channel approach, heavy ion fusion dynamics is investigated for $^{12}\mathrm{C}+^{248}\mathrm{Cm}$ and $^{16}\mathrm{O}+^{244}\mathrm{Pu}$ reactions at energies lying below and near the Coulomb barrier. Coupling of various channels linked with the structure of participants to the relative motion of the collision partners is done by considering a Gaussian type of weight function in the SAGBD model and cross sections are found to be enhanced relative to the calculations obtained by the simple barrier penetration model. In the SAGBD model, the channel coupling effects are calculated in terms of channel coupling parameter ($\ensuremath{\lambda}$) and percentage reduction in the height of apparent fusion barrier with respect to the Coulomb barrier (${V}_{\mathrm{CBRED}}$). The channel coupling parameter estimates the cumulative influence of dominant intrinsic channels, which are responsible for the sub-barrier fusion enhancement. The SAGBD calculations appropriately explain the dynamics of $^{12}\mathrm{C}+^{248}\mathrm{Cm}$ and $^{16}\mathrm{O}+^{244}\mathrm{Pu}$ reactions at energies lying around the Coulomb barrier. The coupled channel analysis of the present reactions is done by using the code ccfull and the coupled channel calculations unambiguously identify the dominant influences of the rotational states up to ${10}^{+}$ spin states of the ground state rotational band of target isotopes in both reactions. In addition, the couplings to higher order deformation, such as ${\ensuremath{\beta}}_{4}$ for target and low lying quantum states of the projectile, are necessarily required to reproduce the experimental data of $^{12}\mathrm{C}+^{248}\mathrm{Cm}$ and $^{16}\mathrm{O}+^{244}\mathrm{Pu}$ reactions. Apart from the fusion analysis, the dynamical cluster-decay model (DCM) is applied to understand the fission dynamics of the $^{260}\mathrm{No}^{*}$ nucleus formed via the above-mentioned reactions. The decay study is carried out at the center-of-mass energies spread, ${E}_{\mathrm{c}.\mathrm{m}.}$ ($\ensuremath{\approx}79$ to 109 MeV) by including the quadrupole deformations (${\ensuremath{\beta}}_{2}$) and optimum orientations (${\ensuremath{\theta}}_{i}^{\mathrm{opt}}$) of the decaying fragments. According to the experimental observation, the noncompound nucleus (nCN) fission component competes with the compound nucleus (CN) fission processes. Consequently, the possibility of nCN contribution is also explored in the decay of the $^{260}\mathrm{No}^{*}$ compound nucleus. With an aim to have a comprehensive analysis of CN and nCN fission mechanisms, the role of the center-of-mass energy (${E}_{\mathrm{c}.\mathrm{m}.}$) and angular momentum ($\ensuremath{\ell}$) is explored in terms of various parameters of DCM such as fragmentation potential, preformation probability, barrier modification, etc.

A higher plant FAD synthetase is fused to an inactivated FAD pyrophosphatase

The riboflavin derivatives FMN and flavin adenine dinucleotide (FAD) are critical cofactors for wide-ranging biological processes across all kingdoms of life. Although it is well established that these flavins can be readily interconverted, in plants, the responsible catalysts and regulatory mechanisms remain poorly understood. Here, we report the cloning and biochemical characterization of an FAD synthetase encoded by the gene At5g03430, which we have designated AtFADS1 (A.thaliana FADS1). The catalytic properties of the FAD synthetase activity are similar to those reported for other FAD synthetases, except that we observed maximum activity with Zn2+ as the associated divalent metal cation. Like human FAD synthetase, AtFADS1 exists as an apparent fusion with an ancestral FAD pyrophosphatase, a feature that is conserved across plants. However, we detected no pyrophosphatase activity with AtFADS1, consistent with an observed loss of a key catalytic residue in higher plant evolutionary history. In contrast, we determined that algal FADS1 retains both FAD synthetase and pyrophosphatase activity. We discuss the implications, including the potential for yet-unstudied biologically relevant noncatalytic functions, and possible evolutionary pressures that have led to the loss of FAD pyrophosphatase activity, yet universal retention of an apparently nonfunctional domain in FADS of land plants.

Cryo-electron tomography with mixed-scale dense neural networks reveals key steps in deployment of Toxoplasma invasion machinery.

Host cell invasion by intracellular, eukaryotic parasites within the phylum Apicomplexa is a remarkable and active process involving the coordinated action of apical organelles and other structures. To date, capturing how these structures interact during invasion has been difficult to observe in detail. Here, we used cryogenic electron tomography to image the apical complex of Toxoplasma gondii tachyzoites under conditions that mimic resting parasites and those primed to invade through stimulation with calcium ionophore. Through the application of mixed-scale dense networks for image processing, we developed a highly efficient pipeline for annotation of tomograms, enabling us to identify and extract densities of relevant subcellular organelles and accurately analyze features in 3-D. The results reveal a dramatic change in the shape of the anteriorly located apical vesicle upon its apparent fusion with a rhoptry that occurs only in the stimulated parasites. We also present information indicating that this vesicle originates from the vesicles that parallel the intraconoidal microtubules and that the latter two structures are linked by a novel tether. We show that a rosette structure previously proposed to be involved in rhoptry secretion is associated with apical vesicles beyond just the most anterior one. This result, suggesting multiple vesicles are primed to enable rhoptry secretion, may shed light on the mechanisms Toxoplasma employs to enable repeated invasion attempts. Using the same approach, we examine Plasmodium falciparum merozoites and show that they too possess an apical vesicle just beneath a rosette, demonstrating evolutionary conservation of this overall subcellular organization.

Enterococcus faecalis alters endo-lysosomal trafficking to replicate and persist within mammalian cells.

Enterococcus faecalis is a frequent opportunistic pathogen of wounds, whose infections are associated with biofilm formation, persistence, and recalcitrance toward treatment. We have previously shown that E. faecalis wound infection persists for at least 7 days. Here we report that viable E. faecalis are present within both immune and non-immune cells at the wound site up to 5 days after infection, raising the prospect that intracellular persistence contributes to chronic E. faecalis infection. Using in vitro keratinocyte and macrophage infection models, we show that E. faecalis becomes internalized and a subpopulation of bacteria can survive and replicate intracellularly. E. faecalis are internalized into keratinocytes primarily via macropinocytosis into single membrane-bound compartments and can persist in late endosomes up to 24 h after infection in the absence of colocalization with the lysosomal protease Cathepsin D or apparent fusion with the lysosome, suggesting that E. faecalis blocks endosomal maturation. Indeed, intracellular E. faecalis infection results in heterotypic intracellular trafficking with partial or absent labelling of E. faecalis-containing compartments with Rab5 and Rab7, small GTPases required for the endosome-lysosome trafficking. In addition, E. faecalis infection results in marked reduction of Rab5 and Rab7 protein levels which may also contribute to attenuated Rab incorporation into E. faecalis-containing compartments. Finally, we demonstrate that intracellular E. faecalis derived from infected keratinocytes are significantly more efficient in reinfecting new keratinocytes. Together, these data suggest that intracellular proliferation of E. faecalis may contribute to its persistence in the face of a robust immune response, providing a primed reservoir of bacteria for subsequent reinfection.

Early outcomes of growth friendly instrumentation in children with Williams syndrome.

Although scoliosis and kyphosis have been associated with Williams Syndrome (WS), no previous literature has reported on surgical treatment for early onset scoliosis (EOS) in WS. The aim of this case series is to report on the outcomes of spine deformity surgery in patients with EOS and WS and any perioperative anesthetic or cardiovascular complications. One multicenter database was queried for all patients with WS who underwent growth-friendly (GF) treatment before age 12 between 2000 and 2017. Demographics, surgical, and growth-friendly data were queried. Radiographs were measured for curve magnitude, T1-T12 length, and T1-S1 length. Seven patients were analyzed (3 males, 4 females). Patients were at a median age of 2.8years at initial surgery with median follow-up 3.6years (range 2.0-12years) after index surgery. The initial surgical treatments were as follows: 2 traditional growing rods (TGR), 2 magnetically controlled growing rods (MCGR), and 3 vertical expandable prosthetic titanium ribs (VEPTR). The median duration of growth-friendly treatment was 5.0years (range, 2.6-10.4years) with a median number of 9 device lengthenings. The median improvement in coronal curve magnitude from preoperative to most recent follow-up was 19° (range, 54°-9°). Three patients have completed GF treatment: one underwent definitive fusion, and two are under observation with apparent spontaneous fusion and retain the original GF implants. No peri-operative anesthetic or cardiovascular complications occurred. Few studies have reported on surgical outcomes in WS patients with EOS. In this case series, 6/7 patients experienced curve improvement with growth-friendly spine instrumentation. This study suggests that growth-friendly instrumentation for severe EOS in WS can be used for control of spinal deformity while allowing for further growth. Associated complications were typical of distraction-based EOS surgical treatment. There were 62 total procedures with general anesthesia, but no perioperative cardiac complications occurred.

T(4;12)(q12;p13) ETV6-rearranged AML without eosinophilia does not involve PDGFRA: relevance for imatinib insensitivity

AbstractAcute myeloid leukemia (AML) with t(4;12)(q12;p13) translocation is rare and often associated with an aggressive clinical course and poor prognosis. Previous reports based on fluorescence in situ hybridization (FISH) analysis have suggested that ETV6::PDGFRA fusions are present in these patients, despite the absence of eosinophilia, which is typically found in other hematopoietic malignancies with PDGFRA-containing fusions. We first detected an ETV6-SCFD2 fusion by targeted RNA sequencing in a patient with t(4;12)(q12;p13) who had been diagnosed with an ETV6-PDGFRA fusion by FISH analysis but failed to respond to imatinib. We then retrospectively identified 4 additional patients with AML and t(4;12)(q12;p13) with apparent ETV6-PDGFRA fusions using chromosome and FISH analysis and applied targeted RNA sequencing to archival material. We again detected rearrangements between ETV6 and non-PDGFRA 4q12 genes, including SCFD2, CHIC2, and GSX2. None of the 3 patients who received imatinib based on the incorrect assumption of an ETV6-PDGFRA fusion responded. Our findings highlight the importance of using a sequencing-based assay to confirm the presence of targetable gene fusions, particularly in genomic regions, such as 4q12, with many clinically relevant genes that are too close to resolve by chromosome or FISH analysis. Finally, combining our data and review of the literature, we show that sequence-confirmed ETV6-PDGFRA fusions are typically found in eosinophilic disorders (3/3 cases), and patients with t(4;12)(q12;p13) without eosinophilia are found to have other 4q12 partners on sequencing (17/17 cases).

Bioabsorbable metal zinc differentially affects mitochondria in vascular endothelial and smooth muscle cells

Zinc is an essential trace element having various structural, catalytic and regulatory interactions with an estimated 3000 proteins. Zinc has drawn recent attention for its use, both as pure metal and alloyed, in arterial stents due to its biodegradability, biocompatibility, and low corrosion rates. Previous studies have demonstrated that zinc metal implants prevent the development of neointimal hyperplasia, which is a common cause of restenosis following coronary intervention. This suppression appears to be smooth muscle cell-specific, as reendothelization of the neointima is not inhibited. To better understand the basis of zinc's differential effects on rat aortic smooth muscle (RASMC) versus endothelial (RAENDO) cells, we conducted a transcriptomic analysis of both cell types following one-week continuous treatment with 5 µM or 50 µM zinc. This analysis indicated that genes whose protein products regulate mitochondrial functions, including oxidative phosphorylation and fusion/fission, are differentially affected by zinc in the two cell types. To better understand this, we performed Seahorse metabolic flux assays and quantitative imaging of mitochondrial networks in both cell types. Zinc treatment differently affected energy metabolism and mitochondrial structure/function in the two cell types. For example, both basal and maximal oxygen consumption rates were increased by zinc in RASMC but not in RAENDO. Zinc treatment increased apparent mitochondrial fusion in RASMC cells but increased mitochondrial fission in RAENDO cells. These results provide some insight into the mechanisms by which zinc treatment differently affects the two cell types and this information is important for understanding the role of zinc treatment in vascular cells and improving its use in biodegradable metal implants.

Color-Coded Imaging of Cancer and Stromal-Cell Interaction in the Pancreatic-Cancer Tumor Microenvironment (TME).

The tumor microenvironment (TME) contains stromal cells in a complex interaction with cancer cells. This relationship has become better understood with the use of fluorescent proteins for in vivo imaging, originally developed by our laboratories. Spectrally-distinct fluorescent proteins can be used for color-coded imaging of the complex interaction of the tumor microenvironment in the living state using cancer cells expressing a fluorescent protein of one color and host mice expressing another-color fluorescent protein. Cancer cells engineered in vitro to express a fluorescent protein were orthotopically implanted into transgenic mice expressing a fluorescent protein of a different color. Confocal microscopy was then used for color-coded imaging of the TME. Color-coded imaging of the TME has enabled us to discover that stromal cells are necessary for metastasis. Patient-derived orthotopic xenograft (PDOX) tumors were labeled by first passaging them orthotopically through transgenic nude mice expressing either green, red, or cyan fluorescent protein in order to label the stromal cells of the tumor (Yang et al., Cancer Res 64:8651-8656, 2004; Yang et al. J Cell Biochem 106: 279-284, 2009). The colored stromal cells become stably associated with the PDOX tumors through multiple passages in transgenic colored nude mice or non-colored nude mice. The fluorescent protein-expressing stromal cells included cancer-associated fibroblasts and tumor-associated macrophages. Color-coded imaging enabled the visualization of apparent fusion of cancer and stromal cells. Color-coded imaging is a powerful tool visualizing the interaction of cancer and stromal cells during cancer progression and treatment.

Color-Coded Imaging of the Tumor Microenvironment (TME) in Human Patient-Derived Orthotopic Xenograft (PDOX) Mouse Models.

The tumor microenvironment (TME) contains stromal cells in a complex interaction with cancer cells. This relationship has become better understood with the use of fluorescent proteins for in vivo imaging, originally developed by our laboratories. Spectrally distinct fluorescent proteins can be used for color-coded imaging of the complex interaction of the tumor microenvironment in the living state using cancer cells expressing a fluorescent protein of one color and host mice expressing another color fluorescent protein. Cancer cells engineered in vitro to express a fluorescent protein were orthotopically implanted into transgenic mice expressing a fluorescent protein of a different color. Confocal microscopy was then used for color-coded imaging of the TME. Color-coded imaging of the TME has enabled us to discover that stromal cells are necessary for metastasis. Patient-derived orthotopic xenograft (PDOX) tumors were labeled by first passaging them orthotopically through transgenic nude mice expressing either green, red, or cyan fluorescent protein in order to label the stromal cells of the tumor. The colored stromal cells become stably associated with the PDOX tumors through multiple passages in transgenic colored nude mice or noncolored nude mice. The fluorescent protein-expressing stromal cells included cancer-associated fibroblasts (CAFs) and tumor-associated macrophages (TAMs). Using this model, specific cancer cell or stromal cell targeting by potential therapeutics can be visualized. Color-coded imaging enabled the visualization of apparent fusion of cancer and stromal cells. Color-coded imaging is a powerful tool visualizing the interaction of cancer and stromal cells during cancer progression and treatment.

Sneaking up on ‘enemies’: alleviating inherent disadvantages in competitive outcomes in a nearly 3-million-year-old palaeocommunity from Florida, USA

Bryozoans offer one of the few systems in which competitive interactions for living space can be studied in the fossil record. Here, we describe the outcome of competitive overgrowths in a 3-million-year-old bryozoan palaeocommunity encrusting shells of the bivalve Anomia simplex from the lower Tamiami Formation in Florida (upper Pliocene, Piacenzian). We found that win–lose overgrowths are more common than stand-offs in interspecific encounters, while stand-offs are more common than win–lose overgrowths in intraspecific encounters. We observed more intraspecific encounters and fewer interspecific interactions than expected under a null hypothesis, suggesting that bryozoans of the same species are likely to be clustered. For some species, intraspecific encounters are more likely to result in the apparent fusion of the two colonies, with the development of rows of kenozooids along the contact edge, probably reflecting relatively low dispersal. We also identified some clear winners and some clear losers. A negative correlation was found between the number of colonies observed and the probability of winning for a species, resulting in a dominance of loser species in the assemblage, a pattern previously described as typical for early colonizers of hard substrates. Our results also confirm the finding of earlier studies that having large zooids and, subordinately, multilayered growth are key traits for success in overgrowth competition, with angle of encounter also having an effect for both poor and good competitors that take advantage of ‘attacking’ colonies of other species from the rear and the flank.

A rare case report of sirenomelia

Mermaid syndrome is a rare congenital fetal anomaly with characteristic feature of complete or partial fusion of lower limbs. Although, this syndrome is incompatible with life due to the association of several congenital defect and malformation. In our case Patient was a 31.3 week. A 1.1 kg, preterm infant of unidentified sex was born, a live born, normally delivered at preterm by a 25-year-old primigravida of lower socioeconomic status. Examination of the baby revealed caudal dysgenesis having apparent fusion of the legs into a single lower limb the most severe form, only a single bone is present,with no indication of legs or feet. There was no identifiable external genitalia and anus. Incidentally the infant died after 45 minutes post birth and we report this case due to their rarity and preterm live birth.

Nematode Autotomy Requires Molting and Entails Tissue Healing without Obvious Regeneration.

Autotomy in C. elegans, which results in the severing of the body into two fragments, has been observed as a response to late larval worm-star formation after exposure to a bacterial surface pathogen. It was found that autotomy can occur in both hermaphroditic and gonochoristic nematode species, and during either the L3 or the L4 molt. Severing was hypothesized to be driven by a ‘balloon-twisting’ mechanism during molting but was found to be independent of lethargus-associated flipping. Extensive healing and apparent tissue fusion were seen at the site of scission. No obvious regeneration of lost body parts was seen in either L4 or adult truncated worms. A variety of mutants defective in processes of cell death, healing, regeneration, responses to damage, stress or pathogens were found to be competent to autotomize. Mutants specifically defective in autotomy have yet to be found. Autotomy may represent a modification of the essential normal process of molting.

Ancestral Eukaryotes Reproduced Asexually, Facilitated by Polyploidy: A Hypothesis.

The notion that eukaryotes are ancestrally sexual has been gaining attention. This idea comes in part from the discovery of sets of "meiosis-specific genes" in the genomes of protists. The existence of these genes has persuaded many that these organisms may be engaging in sex, even though this has gone undetected. The involvement of sex in protists is supported by the view that asexual reproduction results in the accumulation of mutations that would inevitably result in the decline and extinction of such lineages. It is argued that this phenomenon can be obviated by polyploidy and that the "meiosis-specific genes" are used in other processes, including polyploidy control and homologous recombination, independent of meiosis. These phenomena account for the finding that these genes are expressed in cultures devoid of apparent cell fusion events. Hence, it is also proposed that asexual, and not sexual, reproduction is the ancestral condition.

Morphometric evaluation of bone regeneration in segmental mandibular bone defects filled with bovine bone xenografts in a split-mouth rabbit model

BackgroundBovine bone grafts have been widely used in dentistry for guided tissue regeneration and can support new bone formation in direct contact with the graft. The aim of this study was to compare the morphometric and bone density changes after using two different bovine bone graft blocks in segmental osseous defects in the mandible of rabbits following different postoperative periods.Material and methodsCritical size segmental defects were surgically created bilaterally in the jaw of 18 rabbits. The defects were filled with either deproteinized bovine bone mineral with 10% collagen (DBBM-C; BioOss Collagen®), lyophilized bovine medullary bone (LBMB; Orthogen®), or left untreated according to a split-mouth design. Animals were sacrificed after 3 or 6 months of healing. The hemimandibles were scanned ex vivo using a high-resolution (19 μm) microcomputed tomography. Morphometric and bone density parameters were calculated in the region of the defect using CT-Analyser (Bruker). Initial graft blocks were used as baseline.ResultsDBBM-C presented a denser microarchitecture, in comparison to LBMB at baseline. DBBM-C and LBMB grafted regions showed a similar progressive remodeling, with a significant decrease in structure complexity and maintenance of bone volume fraction during the postoperative follow-up periods. Both graft materials showed an enhanced bone replacement and more complex structure compared to untreated defects. The apparent fusion between the graft and host bone was observed only in the defects filled with LBMB.ConclusionLBMB grafts showed a similar behavior as DBBM-C regarding structural remodeling. In LBMB samples, apparent integration between the host bone and the graft was present.

Condensin II subunit NCAPH2 associates with shelterin protein TRF1 and is required for telomere stability.

Condensin II subunits are known to be expressed and localized to interphase nuclei of eukaryotic cells. Although some studies have shown that condensin II likely exerts axial compaction forces, organizes chromosome territories, and has possible transcriptional modulatory functions, the full range of condensin II interphase activities are not known. In particular, it is not known if condensin II interphase activities are generally genome‐wide or if they have additional local activities unique to specific chromosomal structures such as telomeres. Here, we find that NCAPH2 interacts with TRF1 and these two proteins co‐localize at telomeres. Depletion of NCAPH2 leads to ATR‐dependent accumulation of 53BP1 and γH2AX DNA damage foci, including damage specific to telomeres. Furthermore, depletion of NCAPH2 results in a fragile telomere phenotype and apparent sister‐telomere fusions only days after NCAPH2 depletion. Taken together these observations suggest that NCAPH2 promotes telomere stability, possibly through a direct interaction with the TRF1 shelterin component, and prevents telomere dysfunction resulting from impaired DNA replication. Because proper telomere function is essential for chromosome integrity these observations reveal a previously unappreciated function for NCAPH2 in ensuring genome and telomere stability.

Rod Fracture After Apparently Solid Radiographic Fusion in Adult Spinal Deformity Patients

Rod fracture occurs with delayed fusion or pseudarthrosis after adult spinal deformity (ASD) surgery. Rod fracture after apparent radiographic fusion has not been previously investigated. Patients with ASD in a multicenter database were assessed for radiographic fusion by a committee of 3 spinal deformity surgeons. Fusions were rated as bilaterally fused (A), unilaterally fused (B), partially fused (C), or not fused (D). Patients with grade A or B fusion and 2-year follow-up were included. Patients with radiographic fusion were evaluated for subsequent rod fracture. Adjusted analyses were conducted with multiple logistic regression, using backwards-variable selection to a threshold of P<0.2, to assess for associated factors. Of 402 patients with radiographically apparent solid fusion, 9.5% (38) subsequently suffered a broken rod. On multivariate analysis, greater rates of rod fracture were seen among patients of age group 60-69 years (vs. 18-49), body mass index 30-34 and 35+ (vs. <25), stainless-steel rods (vs. titanium), patients with rods ≤5.5 mm (vs. 6.35 mm), and patients with Charlson score 0 (vs. 3+). Of the 38 patients with rod fractures, 18 (47.4%) presented with worsened pain, and 8 (21.1%) required revision at minimum 2-year follow-up. Rod fracture occurred in 9.5% of patients with apparently solid radiographic fusion after ASD surgery. Advanced age, obesity, small diameter rods (5.5 mm), osteotomy, and lower comorbidity burden were significantly associated with rod fracture. Nearly one-half of these patients noted worsening pain, and 21.1% required revision surgery. Instrumentation failure may occur and may be symptomatic even in the setting of apparent fusion on plain radiographs.

Fractures After Removal of Spinal Instrumentation: Revisiting the Stress-Shielding Effect of Instrumentation in Spine Fusion

Many practicing spine surgeons believe that instrumentation can be removed during revision surgery in successful posterolateral or anterior spinal fusions, confirmed by computed tomography and intraoperative exploration. The stress-shielding effect of spinal instrumentation was well described in the late 1980s and 1990s but has not received recent attention. Despite the paucity of recent literature, concepts underlying the biology and biomechanics of the spinal fusion mass remain particularly salient given the increasing incidence of revision spinal fusion surgery. The aim of this study was to highlight a potential complication of instrumentation removal owing to stress shielding of instrumentation on the spinal column and fusion mass. A retrospective review was performed, and a small case series was described. In 3 cases, despite apparent solid fusion demonstrated on preoperative computed tomography and confirmed by intraoperative exploration, new fractures developed after removal of instrumentation. In these cases, fracture occurred at the transition zone between the newly rigid instrumented area and previous fusion. This highlights the relative weakness of the fusion and may be explained by the stress-shielding effect of instrumentation within the fusion mass. Spinal instrumentation revision requires careful consideration, and routine implant removal should not be performed. The presence of a solid fusion on computed tomography and/or at intraoperative exploration may not justify implant removal in these cases. In cases of extension of a fusion, use of a bridging connection to the new implants should be considered. The cases presented demonstrate the consequences of the stress-shielding effect of implants on the spine and fusion mass.

Visualizing the Tumor Microenvironment by Color-coded Imaging in Orthotopic Mouse Models of Cancer.

The tumor microenvironment (TME) contains stromal cells in a complex interaction with cancer cells. This relationship has become better understood with the use of fluorescent proteins for in vivo imaging, originally developed by our laboratories. Spectrally-distinct fluorescent proteins can used for color-coded imaging of the complex interaction of the tumor microenvironment in the living state using cancer cells expressing a fluorescent protein of one color and host mice expressing another-color fluorescent protein. Cancer cells engineered in vitro to express a fluorescent protein were orthotopically implanted into transgenic mice expressing a fluorescent protein of a different color. Confocal microscopy was then used for color-coded imaging of the TME. Color-coded imaging of the TME has enabled us to discover that stromal cells are necessary for metastasis. Patient-derived orthotopic xenograft (PDOX) tumors were labeled by first passaging them orthotopically through transgenic nude mice expressing either green, red, or cyan fluorescent protein in order to label the stromal cells of the tumor. The colored stromal cells become stably associated with the PDOX tumors through multiple passages in transgenic colored mice or non-colored mice. The fluorescent protein-expressing stromal cells included cancer-associated fibroblasts and tumor-associated macrophages. The cancer cells in PDOX models can also be labeled with a telomerase-dependent adenovirus containing the gene for green fluorescent protein. Using this model, specific cancer-cell or stromal-cell targeting by potential therapeutics can be visualized. Color-coded imaging enabled the visualization of apparent fusion of cancer and stromal cells. Color-coded imaging is a powerful tool visualizing the interaction of cancer and stromal cells during cancer progression and treatment.

ViFi: accurate detection of viral integration and mRNA fusion reveals indiscriminate and unregulated transcription in proximal genomic regions in cervical cancer.

The integration of viral sequences into the host genome is an important driver of tumorigenesis in many viral mediated cancers, notably cervical cancer and hepatocellular carcinoma. We present ViFi, a computational method that combines phylogenetic methods with reference-based read mapping to detect viral integrations. In contrast with read-based reference mapping approaches, ViFi is faster, and shows high precision and sensitivity on both simulated and biological data, even when the integrated virus is a novel strain or highly mutated. We applied ViFi to matched genomic and mRNA data from 68 cervical cancer samples from TCGA and found high concordance between the two. Surprisingly, viral integration resulted in a dramatic transcriptional upregulation in all proximal elements, including LINEs and LTRs that are not normally transcribed. This upregulation is highly correlated with the presence of a viral gene fused with a downstream human element. Moreover, genomic rearrangements suggest the formation of apparent circular extrachromosomal (ecDNA) human-viral structures. Our results suggest the presence of apparent small circular fusion viral/human ecDNA, which correlates with indiscriminate and unregulated expression of proximal genomic elements, potentially contributing to the pathogenesis of HPV-associated cervical cancers. ViFi is available at https://github.com/namphuon/ViFi.