Enlarging premaxillary mass: A unique manifestation of extranodal diffuse B-cell lymphoma.

Lymphomas are the most common nonepithelial malignancy in the head and neck region. Among these, non-Hodgkin Lymphoma (NHL) is the most prevalent, and diffuse large B-cell lymphoma (DLBCL) is the most common histologic subtype. NHL is known for its propensity for extranodal involvement, which can affect any anatomical location. The presence of perineural spread is frequently encountered in head and neck malignancies, including lymphomas. We report a case of a 40-year-old male with an enlarging infraorbital facial mass with associated erythema, pain, and paresthesia, which was subsequently found to be extranodal DLBCL with retrograde perineural spread along the infraorbital nerve.

Artificial intelligence-augmented histopathologic review using image analysis to optimize DNA yield from formalin-fixed paraffin-embedded slides.

To achieve minimum DNA input requirements for next-generation sequencing (NGS), pathologists visually estimate macrodissection and slide count decisions. Unfortunately, misestimation may cause tissue waste and increased laboratory costs. We developed an artificial intelligence (AI)-augmented smart pathology review system (SmartPath) to empower pathologists with quantitative metrics for accurately determining tissue extraction parameters. SmartPath uses two deep learning architectures, a U-Net based network for cell segmentation and a multi-field-of-view convolutional network for tumor area segmentation, to extract features from digitized H&E-stained formalin-fixed paraffin-embedded slides. From the segmented tumor area, SmartPath suggests a macrodissection area. To predict DNA yield per slide, the extracted features from within the macrodissection area are correlated with known DNA yields to fit a regularized linear model (R = 0.85). Then, a pathologist-defined target yield divided by the predicted DNA yield per slide gives the number of slides to scrape. Following model development, an internal validation trial was conducted within the Tempus Labs molecular sequencing laboratory. We evaluated our system on 501 clinical colorectal cancer slides, where half received SmartPath-augmented review and half traditional pathologist review. The SmartPath cohort had 25% more DNA yields within a desired target range of 100-2000 ng. The number of extraction attempts was statistically unchanged between cohorts. The SmartPath system recommended fewer slides to scrape for large tissue sections, saving tissue in these cases. Conversely, SmartPath recommended more slides to scrape for samples with scant tissue sections, especially those with degraded DNA, helping prevent costly re-extraction due to insufficient extraction yield. A statistical analysis was performed to measure the impact of covariates on the results, offering insights on how to improve future applications of SmartPath. With these improvements, AI-augmented histopathologic review has the potential to decrease tissue waste, sequencing time, and laboratory costs by optimizing DNA yields, especially for samples with scant tissue and/or degraded DNA.

Arthroscopic Treatment of Osteochondral Lesions of the Talus in a Pediatric Population: A Minimum 2-Year Follow-up.

BACKGROUND: Treatment of osteochondral lesions of the talus (OLTs) in children presents a difficult clinical challenge, with few large series reported. PURPOSE: To evaluate functional and radiographic outcomes for children and adolescents undergoing arthroscopic treatment of symptomatic OLT with a minimum follow-up of 2 years. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: Patients were identified who had symptomatic OLT treated arthroscopically with marrow stimulation techniques. Inclusion criteria were age ≤18 years, symptomatic chronic OLT as the surgical indication, failure of nonoperative treatment, and minimum follow-up of 24 months. Outcome measures included Foot Function Index, American Orthopaedic Foot and Ankle Society Hindfoot Score, Tegner Activity Scale, 36-Item Short Form Health Survey (Short Form-36, v 2), visual analog scale, ankle range of motion, and patient satisfaction survey. Weightbearing radiographs were compared with preoperative radiographs via an ankle arthritis classification system. Magnetic resonance imaging (MRI) was used to evaluate postoperative lesion characteristics per the MOCART scale (magnetic resonance observation of cartilage repair tissue). The size, location, lesion stability, traumatic etiology, skeletal maturity, and length of follow-up were recorded and analyzed through univariate logistic regression. RESULTS: The study group consisted of 22 patients (11 male, 11 female) with a mean age of 14.4 years (range, 8-18 years) and a mean follow-up of 8.3 years (range, 2-27 years). Of 22 patients, 20 were satisfied with the results from surgery and would recommend it to others. Mean follow-up visual analog scale for pain was reported as 2.2 on a 10-point scale, and mean American Orthopaedic Foot and Ankle Society score at follow-up was 86.6. Mean postoperative Foot Function Index scores for the study group were as follows: pain, 17.1; disability, 16.5; activity, 4.7; and overall, 38.7. Mean Short Form-36 physical component score was 50.7. Postoperative radiographs indicated a van Dijk osteoarthritis grade of 0 in 56%, I in 38%, II in 6%, and III in 0%. Postoperative MRI MOCART scores showed complete filling of the cartilage in 27% of cases, complete graft integration in 22%, and intact repair surface in 22%, with a mean MOCART score of 48.0. No correlation was found between radiographic and MRI findings and clinical outcomes. None of the prognostic factors were significantly associated with patient satisfaction, progression of arthritis, or MOCART scores. CONCLUSION: Arthroscopic treatment of symptomatic OLT in adolescent patients (≤18 years) demonstrated high functional outcomes, high clinical satisfaction rates, and minimal radiographic osteoarthritic progression despite low MOCART scores.

The Peptidisc, a simple method for stabilizing membrane proteins in detergent-free solution.

Membrane proteins are difficult to work with due to their insolubility in aqueous solution and quite often their poor stability in detergent micelles. Here, we present the peptidisc for their facile capture into water-soluble particles. Unlike the nanodisc, which requires scaffold proteins of different lengths and precise amounts of matching lipids, reconstitution of detergent solubilized proteins in peptidisc only requires a short amphipathic bi-helical peptide (NSPr) and no extra lipids. Multiple copies of the peptide wrap around to shield the membrane-exposed part of the target protein. We demonstrate the effectiveness of this 'one size fits all' method using five different membrane protein assemblies (MalFGK2, FhuA, SecYEG, OmpF, BRC) during 'on-column', 'in-gel', and 'on-bead' reconstitution embedded within the membrane protein purification protocol. The peptidisc method is rapid and cost-effective, and it may emerge as a universal tool for high-throughput stabilization of membrane proteins to advance modern biological studies.

Selectively targeting the dimerization interface of human androgen receptor with small-molecules to treat castration-resistant prostate cancer.

Prostate cancer (PCa) is a leading cause of death for men in North America. The androgen receptor (AR) - a hormone inducible transcription factor - drives expression of tumor promoting genes and represents an important therapeutic target in PCa. The AR is activated by steroid recruitment to its ligand binding domain (LBD), followed by receptor nuclear translocation and dimerization via the DNA binding domain (DBD). Clinically used small molecules interfere with steroid recruitment and prevent AR-driven tumor growth, but are rendered ineffective by emergence of LBD mutations or expression of constitutively active variants, such as ARV7, that lack the LBD. Both drug-resistance mechanisms confound treatment of this 'castration resistant' stage of PCa (CRPC), characterized by return of AR signalling. Here, we employ computer-aided drug-design to develop small molecules that block the AR-DBD dimerization interface, an attractive target given its role in AR activation and independence from the LBD. Virtual screening on the AR-DBD structure led to development of prototypical compounds that block AR dimerization, inhibiting AR-transcriptional activity through a LBD-independent mechanism. Such inhibitors may potentially circumvent AR-dependent resistance mechanisms and directly target CRPC tumor growth.

Comparing discrete distributions when the sample space is smal / Comparando las distribuciones discretas cuando el espacio muestral es pequeño

This paper describes two new methods for comparing two independent, discrete distributions, when the sample space is small, using an extension of the Storer-Kim method for comparing independent binomials. These methods are relevant, for example, when comparing groups based on a Likert scale, which was the motivation for the paper. In essence, the goal is to test the hypothesis that the cell probabilities associated with two independent multinomial distributions are equal. Both a global test and a multiple comparison procedure are proposed. The small-sample properties of both methods are compared to four other techniques via simulations: Cliff's generalization of the Wilcoxon-Mann-Whitney test that effectively deals with heteroscedasticity and tied values, Yuen's test based on trimmed means, Welch's test and Student's t test. For the simulations, data were generated from beta-binomial distributions. Both symmetric and skewed distributions were used. The sample space consisted of the integers 0(1)4 or 0(1)10. For the global test that is proposed, when testing at the 0.05 level, simulation estimates of the actual Type I error probability ranged between 0.043 and 0.059. For the new multiple comparison procedure, the estimated family wise error rate ranged between 0.031 and 0.054 for the sample space 0(1)4. But for 0(1)10, the estimates dropped as low as 0.016 in some situations. Given the goal of comparing means, Student's t is well known to have practical problems when distributions differ. Similar problems are found here among the situations considered. No single method dominates in terms of power, as would be expected, because different methods are sensitive to different features of the distributions being compared. But in general, one of the new methods tends to have relatively good power based on both simulations and experience with data from actual studies. If, however, there is explicit interest in comparing means, rather than comparing the cell probabilities, Welch's test was found to perform well. The new methods are illustrated using data from the Well-Elderly Study where the goal is to compare groups in terms of depression and the strategies used for dealing with stress.

En este artículo se describen dos nuevos métodos para comparar dos distribuciones discretas independientes, cuando el espacio muestral es pequeño, usando una extensión del método Storer-Kim para comparar binomios independientes. Estos métodos son relevantes, por ejemplo, cuando se comparan grupos basados en una escala Likert, la cual motivó la escritura del artículo. En esencia, el objetivo es evaluar la hipótesis de que las probabilidades de células asociadas con dos distribuciones multinominales independientes son iguales. Se propone una prueba global y un procedimiento de comparación múltiple. Las propiedades de las muestras pequeñas de ambos métodos fueron comparadas con otras cuatro técnicas a través de simulaciones: generalización de Cliff de la prueba de Wilcoxon-Mann-Whitney que trata eficazmente con heteroscedasticidad y valores vinculados, la prueba de Yuen basada en medias truncadas, la prueba de Welch y la prueba t de Student. Para las simulaciones, los datos se generaron a partir de distribuciones beta-binomiales. Se utilizaron distribuciones tanto simétricas como asimétricas. El espacio muestral consistió en los enteros 0(1)4 o 0(1)10. Para la prueba global que se propone, cuando se evaluó al nivel de 0.05, la simulación estimó la probabilidad del error tipo I osciló entre 0.043 y 0.059. Para el nuevo procedimiento de comparación múltiple, la tasa de error estimada oscilaba entre 0.031 y 0.054 para el espacio de la muestra 0(1)4. Pero para 0(1)10, las estimaciones fueron tan bajas como 0.016 en algunas situaciones. Teniendo en cuenta el objetivo de la comparación de medias, la prueba t de Student es bien conocida por tener problemas prácticos cuando distribuciones difieren. Problemas similares se encuentraron entre las situaciones consideradas. No existe un único método que domina en términos de poder, como sería de esperar, debido a que los diferentes métodos son sensibles a las diferentes características de las distribuciones que son comparadas. Pero en general, uno de los nuevos métodos tiende a tener relativamente buen poder basado tanto en simulaciones y la experiencia con los datos de estudios reales. Si, sin embargo, existe un interés explícito en comparar medias, en lugar de comparar las probabilidades de celda, la prueba de Welch se encuentra que tiene un buen desempeño. Los nuevos métodos se ilustran usando datos del estudio Well-Elderly donde el objetivo es comparar los grupos en cuanto a la depresión y las estrategias utilizadas para hacer frente al estrés.

Complications in ankle and foot arthroscopy.

Arthroscopy has many potential complications, whether it is done in the ankle, shoulder, knee, or other joints. Foot and ankle arthroscopy has progressed significantly since its beginning by Burman in 1931. Over the past 2 decades, arthroscopy equipment and instrumentation has improved and newer techniques have been developed. A heightened interest in foot and ankle arthroscopy has grown as diagnostic and imaging capabilities have improved. As the number of arthroscopic procedures of the foot and ankle has increased, so has the opportunity for significant complications.

Attenuation of acute graft-versus-host disease in the absence of the transcription factor RORγt.

Graft-versus-host disease (GVHD) remains the most significant complication after allogeneic stem cell transplantation. Previously, acute GVHD had been considered to be mediated predominantly by Th1-polarized T cells. Recently, investigators have identified a second proinflammatory lineage of T cells termed Th17 that is critically dependent on the transcription factor retinoic acid-related orphan receptor (ROR)γt. In this study, we have evaluated the role of Th17 cells in murine acute GVHD by infusing donor T cells lacking RORC and as a consequence the isoform RORγt. Recipients given donor CD4(+) and CD8(+) T cells lacking RORC had significantly attenuated acute GVHD and markedly decreased tissue pathology in the colon, liver, and lung. Using a clinically relevant haploidentical murine transplantation model, we showed that RORC(-/-) CD4(+) T cells alone diminished the severity and lethality of acute GVHD. This was not found when CD4(+) T cells from RORC(-/-) mice were given to completely mismatched BALB/c mice, and it was correlated with absolute differences in the generation of TNF in the colon after transplant. Thus, CD4(+) T cell expression of RORC is important in the pathogenesis of acute GVHD.

Separation of graft-versus-host disease from graft-versus-leukemia responses by targeting CC-chemokine receptor 7 on donor T cells.

CC-chemokine receptor 7 (CCR7) is expressed on the surface of naive T cells, and plays a critical role in their movement into secondary lymphoid tissue. Here, we show that murine T cells lacking CCR7 (CCR7(-/-)) generate attenuated graft-versus-host disease (GVHD) responses compared with wild-type (WT) cells, with the difference varying inversely with the degree of major histocompatibility complex (MHC) disparity between the donor and recipient. CCR7(-/-) T cells exhibited an impaired ability to traffic to recipient lymph nodes, with an increased capacity to home to the spleen. CCR7(-/-) T cells, however, demonstrated a reduced ability to undergo in vivo expansion in the spleen due to impaired interactions with splenic antigen-presenting cells. On a cellular level, CCR7(-/-) T cells were functionally competent, demonstrating a normal in vitro proliferative capacity and a preserved ability to produce inflammatory cytokines. Importantly, CCR7(-/-) T cells were capable of generating robust graft-versus-leukemia (GVL) responses in vivo, as well as complete donor T-cell reconstitution. CCR7(-/-) regulatory T cells were able to protect against lethal GVHD when administered before WT conventional T cells. Our data suggest that CCR7 inhibition in the early posttransplantation period may represent a feasible new therapeutic approach for acute GVHD attenuation without compromising GVL responses.

Healed cuff repairs impart normal shoulder scores in those 65 years of age and older.

BACKGROUND: It is unclear whether repaired rotator cuffs heal in older patients and whether the function in those shoulders compares with those of similarly aged patients with untreated tears. QUESTIONS/PURPOSE: We questioned whether, in patients 65 years of age and older, shoulders with rotator cuff repairs that remained intact would have Simple Shoulder Test (SST) scores and Constant scores similar to those of untreated individuals with intact rotator cuffs. METHODS: We retrospectively reviewed 39 patients (42 shoulders) 65 years of age and older in whom 42 full-thickness rotator cuff tears were repaired with a mini open technique. All patients completed SST and Constant scores 12 to 60 months postoperatively; all patients also had ultrasound at those times to assess the status of the repair. These findings were compared with 200 untreated similarly aged shoulders assessed in the same fashion. RESULTS: Shoulders with healed repairs (33 of 42) had similar mean SST scores and Constant scores to those in untreated shoulders with intact rotator cuffs. Those with healed repairs also had higher SST and Constant scores than those with unhealed repairs. Finally, shoulders with healed repairs had higher SST and Constant scores than those with untreated tears. CONCLUSIONS: When rotator cuffs healed the function was comparable to that of similarly-aged patients without tears and better than that of patients with untreated tears. LEVEL OF EVIDENCE: Level III, therapeutic study. See Guidelines for Authors for a complete description of levels of evidence.

In vitro-differentiated TH17 cells mediate lethal acute graft-versus-host disease with severe cutaneous and pulmonary pathologic manifestations.

The morbidity and mortality associated with graft-host-disease (GVHD) is a significant obstacle to the greater use of allogeneic stem cell transplantation. Donor T cells that predominantly differentiate into TH1/Tc1 T cells and generate pro-inflammatory cytokines such as interferon-gamma (IFN-gamma) mediate GVHD. Although numerous studies have described a pathogenic role for IFN-gamma, multiple reports have demonstrated that the lack of IFN-gamma paradoxically exacerbated GVHD lethality. This has led to speculation that another subset of T cells may significantly contribute to GVHD mortality. Several groups have demonstrated a new lineage of CD4+ T helper cell development distinct from TH1 or TH2 differentiation. This lineage is characterized by production of interleukin (IL)-17A, IL-17F, IL-22, and IL-21 and has been termed TH17 cells. Here, we demonstrate that a highly purified population of TH17 cells is capable of inducing lethal GVHD, hallmarked by extensive pathologic cutaneous and pulmonary lesions. Upon transfer, these cells migrate to and expand in GVHD target organs and secondary lymphoid tissues. Finally, we demonstrate differential roles for tumor necrosis factor-alpha (TNF-alpha) and IL-17A in the clinical manifestations of GVHD induced by TH17 cells. Our studies demonstrate that cells other than TH1/Tc1 can mediate acute GVHD.

The biology and therapeutic potential of natural regulatory T-cells in the bone marrow transplant setting.

Graft-versus-host disease (GVHD) remains a major barrier to the long-term success of allogeneic hematopoietic stem-cell transplantation for hematologic malignancies. This has prompted a great deal of interest in the development of alternative GVHD management strategies such as the adoptive transfer of lymphocytes with immunosuppressant activity. This review will focus on the biology of thymically derived regulatory T-cells (Tregs) and their potential therapeutic role in human bone marrow transplantation. A discussion of their basic suppressive mechanisms and in vivo trafficking patterns will be presented, along with an overview of important preclinical studies using murine GVHD/tumor models. Currently available clinical data will also be discussed, including the contribution of Tregs to various transplant outcome parameters such as GVHD incidence and malignancy relapse rates.

B-cell studies in chronic ethanol mice.

Chronic alcohol abuse leads to multiple defects in the immune system, leading to an increased risk of infectious disease and malignancy. Immune lesions encompass both the innate and adaptive arms and include deficiencies in the B-cell compartment. Long-term alcoholics exhibit loss of B cells in the periphery and diminished ability to generate protective antibodies. To better mimic the chronic alcoholic patient, our group has used an ethanol-in-drinking-water mouse model. Mice consuming alcohol in this manner progressively develop a range of immune abnormalities, including defects in humoral immunity. To document and explore B-cell lesions in ethanol-consuming mice, our laboratory has used a broad panel of technologies. These include protocols to define the physical state of B cells in the bone marrow and periphery, in vitro approaches to test B-cell activation potential and in vivo experiments to document the humoral competence of the host. These key techniques are detailed in the present chapter.