Somatic and intergenerational G4C2 hexanucleotide repeat instability in a human C9orf72 knock-in mouse model.

Expansion of a G4C2 repeat in the C9orf72 gene is associated with familial Amyotrophic Lateral Sclerosis (ALS) and Frontotemporal Dementia (FTD). To investigate the underlying mechanisms of repeat instability, which occurs both somatically and intergenerationally, we created a novel mouse model of familial ALS/FTD that harbors 96 copies of G4C2 repeats at a humanized C9orf72 locus. In mouse embryonic stem cells, we observed two modes of repeat expansion. First, we noted minor increases in repeat length per expansion event, which was dependent on a mismatch repair pathway protein Msh2. Second, we found major increases in repeat length per event when a DNA double- or single-strand break (DSB/SSB) was artificially introduced proximal to the repeats, and which was dependent on the homology-directed repair (HDR) pathway. In mice, the first mode primarily drove somatic repeat expansion. Major changes in repeat length, including expansion, were observed when SSB was introduced in one-cell embryos, or intergenerationally without DSB/SSB introduction if G4C2 repeats exceeded 400 copies, although spontaneous HDR-mediated expansion has yet to be identified. These findings provide a novel strategy to model repeat expansion in a non-human genome and offer insights into the mechanism behind C9orf72 G4C2 repeat instability.

Influence of eight debridement techniques on three different titanium surfaces: A laboratory study.

OBJECTIVES: Debridement methods may damage implant surfaces. This in vitro study investigated eight debridement protocols across three implant surfaces to assess both biofilm removal and surface alterations. MATERIAL AND METHODS: One hundred sixty commercially pure titanium discs were treated to simulate commercially available titanium implant surfaces-smooth, abraded and abraded and etched. Following inoculation with whole human saliva to create a mixed species biofilm, the surfaces were treated with eight debridement methods currently used for clinical peri-implantitis (n = 10). This included air abrasion using powders of glycine, sodium bicarbonate and calcium carbonate; conventional mechanical methods-piezoelectric scaler, carbon and stainless steel scalers; and a chemical protocol using 40% citric acid. Following treatment, remaining biofilm was analysed using scanning electron microscopy and crystal violet assays. For statistical analysis, ANOVA was applied (p < 0.05). RESULTS: All debridement techniques resulted in greater than 80% reduction in biofilm compared with baseline, irrespective of the surface type. Glycine powder delivered through an air polishing system eliminated the most biofilm. Mechanical instruments were the least effective at eliminating biofilm across all surfaces and caused the greatest surface alterations. Citric acid was comparable with mechanical debridement instruments in terms of biofilm removal efficacy. Titanium surfaces were least affected by air abrasion protocols and most affected by mechanical methods. CONCLUSIONS: Mechanical protocols for non-surgical debridement should be approached with caution. Glycine powder in an air polisher and 40% citric acid application both gave minimal alterations across all implant surfaces, with glycine the superior method in terms of biofilm removal.

Fetal liver and lung volume index of neonatal survival with congenital diaphragmatic hernia.

BACKGROUND: Magnetic resonance imaging (MRI) assesses pulmonary hypoplasia in fetal congenital diaphragmatic hernia (CDH). Neonatal mortality may occur with CDH. OBJECTIVE: To quantify MRI parameters associated with neonatal survival in fetuses with isolated CDH. MATERIALS AND METHODS: Fetal MRI for assessing CDH included region of interest (ROI) measurements for total lung volume (TLV), herniated liver volume, herniated other organ volume and predicted lung volume. Ratios of observed lung volume and liver up volume to predicted lung volume (observed to predicted TLV, percentage of the thorax occupied by liver) were calculated and compared to neonatal outcomes. Analyses included Wilcoxon rank sum test, multivariate logistic regression and receiver operating characteristic (ROC) curves. RESULTS: Of 61 studies, the median observed to predicted TLV was 0.25 in survivors and 0.16 in non-survivors (P=0.001) with CDH. The median percentage of the thorax occupied by liver was 0.02 in survivors and 0.22 in non-survivors (P<0.001). The association of observed to predicted TLV and percentage of the thorax occupied by liver with survival for gestational age (GA) >28 weeks was greater compared to GA ≤28 weeks. The ROC analysis demonstrated an area under the curve of 0.96 (95% confidence interval 0.91-1.00) for the combined observed to predicted TLV, percentage of the thorax occupied by liver and GA. CONCLUSION: The percentage of the thorax occupied by liver and observed to predicted TLV was predictive of neonatal survival in fetuses with CDH.

MRI of the Placenta Accreta Spectrum (PAS) Disorder: Radiomics Analysis Correlates With Surgical and Pathological Outcome.

BACKGROUND: Placenta accreta spectrum (PAS) in women with previous cesarean delivery has become increasingly prevalent. Depending on the severity, patient management may involve cesarean hysterectomy. PURPOSE: To investigate textural analyses as the radiomics in MRI of the placenta in predicting the PAS requiring cesarean hysterectomy in a high-risk population. STUDY TYPE: Retrospective. POPULATION: Sixty-two women with prior cesarean delivery referred for MRI because of sonographic suspicion for PAS. FIELD STRENGTH/SEQUENCE: 1.5T with T1 W, T2 W, and diffusion-weighted imaging (DWI). ASSESSMENT: Two reviewers independently evaluated MR images based on five established PAS variables. Placental regions of interest (ROIs) were generated on T2 W, DWI, and an apparent diffusion coefficient (ADC) map, based on definitions of areas of placenta in proximity to and remote from previous surgical incision sites. STATISTICAL TESTS: Reader agreement was assessed by simple kappa and prevalence adjusted bias adjusted kappa (PABAK). T-tests and chi-square analyses between the primary outcome (hysterectomy vs. no hysterectomy) were performed. Thirteen Haralick texture features calculated from gray-level co-occurrence matrixes were extracted from manually drawn placental ROIs within each of three MR acquisitions. Univariate and multivariable logistic regression were used to assess the association with cesarean hysterectomy. RESULTS: Of 62 pregnancies at risk for PAS, 40 required cesarean hysterectomy (65%), with excellent correlation between need for hysterectomy and pathology confirmation of PAS in the hysterectomy specimen [κ = 0.82 (0.62, 1)]. Reader agreement was fair to moderate. Of the 13 Haralick variables within each of three acquisition groups, significant differences (P < 0.05) were seen in 22 of 39 parameters comparing placental ROIs in proximity to incision scar(s) to those ROIs remote from scar. A stepwise selection algorithm indicated that the combination of T2 W Fcm.sum.var , ADC Fcm.diff.entr , and DWI Fcm.energy gave the highest leave-one-out-AUC of 0.80 (0.68, 0.91). DATA CONCLUSION: Assessment of PAS severity is subjective and dependent on radiologist expertise. We identified textural features on placental MR images in the region of the prior uterine scar that differentiated pregnancies requiring cesarean hysterectomy based on clinical suspicion of PAS from those that did not, suggesting predictive capabilities of these objective radiomics features. LEVEL OF EVIDENCE: 3 Technical Efficacy Stage: 1 J. Magn. Reson. Imaging 2020;51:936-946.

Placenta Accreta Spectrum: Correlation of MRI Parameters With Pathologic and Surgical Outcomes of High-Risk Pregnancies.

OBJECTIVE. The purpose of this study is to determine whether MRI parameters of placenta accreta spectrum correlate with pathologic and surgical outcomes in high-risk pregnancies. MATERIALS AND METHODS. This retrospective study evaluated second- and third-trimester pregnancies assessed by MRI from 2007 to the present. Women were included in the study if placental invasion was suspected on the basis of a clinical history of cesarean delivery, ultrasound findings, or both. MR images were reviewed by an experienced radiologist who was blinded to the clinical outcomes. Eighteen MRI parameters were assessed and compared with four clinical outcomes: surgical impression of invasion, need for cesarean hysterectomy, pathologic findings, and need for blood transfusion. RESULTS. Of 64 women, 43 required cesarean hysterectomy, 20 underwent cesarean delivery, and one delivered vaginally. There was no statistical difference among the women in terms of maternal age, gestational age, or the number of prior cesarean deliveries. Eight of the 18 MRI parameters assessed showed statistical significance. The five variables with the highest odds ratios were bulge (7.432), placenta previa (7.283), low-attenuation T2 linear bands (5.985), placental heterogeneity near the scar (4.384), and fibrin deposition (4.322), with additional significant variables including interruption of the bladder-serosa interface, the radiologist's interpretation of invasion, and the largest dimension of invasion. Some previously described parameters, such as the degree of maternal pelvic vascularity, were not statistically significant. CONCLUSION. MRI parameters are associated with placental invasion and correlate with the need for cesarean hysterectomy, as well as pathologic and surgical impressions of invasion. From these parameters, an organized template can be created to standardize reporting of placental invasion.

Liver Injury in Hemolysis, Elevated Liver Enzymes, and Low Platelets Syndrome Measured by Diffusion-Weighted Magnetic Resonance Imaging.

OBJECTIVE: The objective of this study was to evaluate acute liver injury (ALI) detected by diffusion-weighted magnetic resonance imaging (MRI) and the associated laboratory findings in women with hemolysis, elevated liver enzymes, and low platelet (HELLP) syndrome. STUDY DESIGN: This was a retrospective, observational study of women with HELLP syndrome defined by serum aspartate aminotransferase (AST) levels ≥100 U/L and thrombocytopenia with platelets ≤100,000/µL. All women underwent MRI postpartum including diffusion-weighted imaging to estimate the volume of ALI with reconstructed apparent diffusion coefficient (ADC) maps. The ADC map and the volume of ALI were compared with laboratory abnormalities by Spearman's correlation analysis. RESULTS: From March 2013 through August 2015, 16 women with HELLP syndrome underwent MRI, and of these, 14 (88%) women had areas of increased signal intensity suggestive of ALI. Their median (range) maximum AST level was 262 (140-1,958) IU/L, and at the time of MRI, AST was 103 (36-1,426) IU/L. Both of these AST levels significantly correlated with ADC map as well as the volume of ALI (both p-values <0.001). CONCLUSION: Women with HELLP syndrome frequently exhibited areas of abnormal diffusion in the liver on diffusion-weighted MRI, suggestive of ALI. The extent of liver injury was significantly correlated with serum AST.

The usefulness of gestational sac and placental sonographic morphology in differentiating between second-trimester tubal and abdominal pregnancy: case report and a review of literature.

Second-trimester tubal pregnancy is rarely encountered and published cases are limited. There are no established sonographic criteria to differentiate it from abdominal pregnancy; however, differences in their medical management make this distinction important. We report a case of a 14-week 5-day tubal pregnancy that presented near rupture. Sonography demonstrated many overlapping features with abdominal pregnancy. In addition, the extrauterine gestational sac was rounded and well defined and the placenta was crescent-shaped. We propose that these sonographic features represent a second-trimester tubal pregnancy rather than an abdominal pregnancy.

Interdisciplinary diagnosis and treatment planning for Class III malocclusion and determining the appropriate anterior tooth positions for individual patients.

Patients presenting with Class III malocclusion often present significant challenges for the orthodontist and restorative clinician. The characteristic anterior crossbite is commonly associated with a maxillo-mandibular skeletal base discrepancy leading to both functional and esthetic issues. Three potential incisal tooth positions are discussed using clinical examples including implants.

Multimodal depiction of a rare immature gastric teratoma from fetus to infant.

Immature gastric teratomas are rare gastrointestinal tumors. Presented here are imaging findings of this neoplasm which was first seen via antenatal ultrasound. Subsequent fetal magnetic resonance imaging demonstrated a partially calcified mass that contained areas of diffusion restriction. Meconium pseudocyst was originally entertained as a differential consideration. Follow-up computed tomography and upper gastrointestinal fluoroscopy after delivery revealed a bilobed mass that was at least partially endogastric. Resection was performed, and the diagnosis was uncovered via histologic analysis.

Diagnostic Evaluation of Chronic Pelvic Pain.

Chronic pelvic pain can result from various intra- and extra-pelvic etiologies. Although patient history and physical examination may narrow the differential diagnosis, frequently, the different etiologies have overlapping presentations. Imaging examinations such as US and/or MR imaging may help delineate the cause of pain, particularly when related to intra-pelvic organs, pelvic floor dysfunction or prolapse, synthetic material such as pelvic mesh or slings, and in some cases of neuropathic pain. Etiologies of neuropathic pain can also be assessed with non-imaging tests such as nerve conduction studies, electromyography, and testing of sacral reflexes.

Biochemical and biomechanical properties of the pacemaking sinoatrial node extracellular matrix are distinct from contractile left ventricular matrix.

Extracellular matrix plays a role in differentiation and phenotype development of its resident cells. Although cardiac extracellular matrix from the contractile tissues has been studied and utilized in tissue engineering, extracellular matrix properties of the pacemaking sinoatrial node are largely unknown. In this study, the biomechanical properties and biochemical composition and distribution of extracellular matrix in the sinoatrial node were investigated relative to the left ventricle. Extracellular matrix of the sinoatrial node was found to be overall stiffer than that of the left ventricle and highly heterogeneous with interstitial regions composed of predominantly fibrillar collagens and rich in elastin. The extracellular matrix protein distribution suggests that resident pacemaking cardiomyocytes are enclosed in fibrillar collagens that can withstand greater tensile strength while the surrounding elastin-rich regions may undergo deformation to reduce the mechanical strain in these cells. Moreover, basement membrane-associated adhesion proteins that are ligands for integrins were of low abundance in the sinoatrial node, which may decrease force transduction in the pacemaking cardiomyocytes. In contrast to extracellular matrix of the left ventricle, extracellular matrix of the sinoatrial node may reduce mechanical strain and force transduction in pacemaking cardiomyocytes. These findings provide the criteria for a suitable matrix scaffold for engineering biopacemakers.

A novel osteotropic biomaterial OG-PLG: Synthesis and in vitro release.

Statins (e.g., simvastatin) have shown to induce expression of the bone morphogenic protein-2 gene in bone cells, but they are not used clinically because of a lack of a suitable delivery device. The overall objective is to develop optimized statin delivery devices for bone regeneration. The specific objective was to determine the effect of grafting statins to biodegradable poly[lactide-co-glycolide] (PLG) on release kinetics. Simvastatin was grafted to PLG (OG-PLG) and characterized using contact-angle measurements, attenuated total reflectance-Fourier transform infrared, and ultraviolet-visible spectroscopy to determine success of the synthesis. An ultraviolet-visible assay for measuring release of statins and degraded OG-PLG in media was also developed. In vitro release studies using films and scaffolds made with PLG, PLG blended with simvastatin (PLG + Sim), and OG-PLG (simvastatin grafted to PLG) blended into PLG at different concentrations showed that release rate of OG-PLG from films was significantly greater than that of PLG + Sim. However, release rate from scaffolds showed PLG + Sim to be significantly higher than that of OG-PLG. The diffusion-controlled release kinetics of simvastatin from PLG + Sim seems to be more heavily affected by device morphology, whereas the degradation-controlled release kinetics seem to be less affected. In short, release kinetics can be modulated by grafting statins to PLG.

A novel osteotropic biomaterial OG-PLG: in vitro efficacy.

Previously, a novel osteotropic biomaterial, OG-PLG [simvastatin grafted to poly(lactide-co-glycolide), PLG], was synthesized and shown to have degradation-controlled release kinetics. The objective here was to determine the effect of grafting statins to PLG on bone regeneration in vitro. Rat bone marrow cells were stimulated in vitro with simvastatin dissolved in media, saponified simvastatin dissolved in media, simvastatin released through diffusion from emulsion freeze-dried scaffolds, and OG-PLG. Unstimulated cultures and cultures stimulated with dexamethasone were used as negative and positive controls, respectively. In vitro bone formation was assessed using the alkaline phosphatase (ALP) and von Kossa assays at different times up to 16 days. ALP analysis revealed that saponified simvastatin at 10(-7)M and OG-PLG significantly increased ALP expression at various time points. von Kossa assay showed that simvastatin, saponified simvastatin, and OG-PLG significantly enhanced mineralization, with the effect from OG-PLG being the most significant. In short, OG-PLG significantly enhanced in vitro bone cell mineralization beyond the effect of simvastatin or saponified simvastatin dissolved in media and simvastatin released via diffusion from scaffolds.

Th1 cytokines in oral lichen planus.

BACKGROUND: Cell-mediated immune responses in oral lichen planus (OLP) may be regulated by cytokines and their receptors. METHODS: In situ cytokine expression and in vitro cytokine secretion in OLP were determined by immunohistochemistry and ELISA. RESULTS: The majority of subepithelial and intraepithelial mononuclear cells in OLP were CD8+. In some cases, intraepithelial CD8+ cells were adjacent to degenerating keratinocytes. CD4+ cells were observed mainly in the deep lamina propria with occasional CD4+ cells close to basal keratinocytes. Mononuclear cells expressed IFN-gamma in the superficial lamina propria and TNF-alpha adjacent to basal keratinocytes. Basal keratinocytes expressed TNF-alpha as a continuous band. TNF R1 was expressed by mononuclear cells and basal and suprabasal keratinocytes. There was variable expression of TGF-beta1 in the subepithelial infiltrate while all intraepithelial mononuclear cells were TGF-beta1-. Keratinocytes in OLP stained weakly for TGF-beta1. Unstimulated OLP lesional T cells secreted IFN-gamma in vitro. TNF-alpha stimulation down-regulated IFN-gamma secretion and up-regulated TNF-alpha secretion. IL-4, IL-10 and TGF-beta1 secretion were not detected. CONCLUSIONS: These data suggest the development of a T helper 1 immune response that may promote CD8+ cytotoxic T-cell activity in OLP.