The N6-methyladenosine Epitranscriptomic Landscape of Lung Adenocarcinoma.

Comprehensive m6A epitranscriptome profiling of primary tumors remains largely uncharted. Here, we profiled the m6A epitranscriptome of 10 non-neoplastic lung (NL) tissues and 51 lung adenocarcinoma (LUAD) tumors, integrating the corresponding transcriptome, proteome and extensive clinical annotations. We identified distinct clusters and genes that were exclusively linked to disease progression through m6A modifications. In comparison with NL tissues, we identified 430 transcripts to be hypo-methylated and 222 to be hyper-methylated in tumors. Among these genes, EML4 emerged as a novel metastatic driver, displaying significant hyper-methylation in tumors. m6A modification promoted the translation of EML4, leading to its widespread overexpression in primary tumors. Functionally, EML4 modulated cytoskeleton dynamics through interacting with ARPC1A, enhancing lamellipodia formation, cellular motility, local invasion, and metastasis. Clinically, high EML4 protein abundance correlated with features of metastasis. METTL3 small molecule inhibitor markedly diminished both EML4 m6A and protein abundance, and efficiently suppressed lung metastases in vivo.

Induction of IFIT1/IFIT3 and inhibition of Bcl-2 orchestrate the treatment of myeloma and leukemia via pyroptosis.

Induction of pyroptosis is proposed as a promising strategy for the treatment of hematological malignancies, but little is known. In the present study, we find clioquinol (CLQ), an anti-parasitic drug, induces striking myeloma and leukemia cell pyroptosis on a drug screen. RNA sequencing reveals that the interferon-inducible genes IFIT1 and IFIT3 are markedly upregulated and are essential for CLQ-induced GSDME activation and cell pyroptosis. Specifically, IFIT1 and IFIT3 form a complex with BAX and N-GSDME therefore directing N-GSDME translocalization to mitochondria and increasing mitochondrial membrane permeabilization and triggering pyroptosis. Furthermore, venetoclax, an activator of BAX and an inhibitor of Bcl-2, displays strikingly synergistic effects with CLQ against leukemia and myeloma via pyroptosis. This study thus reveals a novel mechanism for mitochondrial GSDME in pyroptosis and it also illustrates that induction of IFIT1/T3 and inhibition of Bcl-2 orchestrate the treatment of leukemia and myeloma via pyroptosis.

Six-year follow-up and subgroup analyses of a phase 2 trial of venetoclax for del(17p) chronic lymphocytic leukemia.

ABSTRACT: Chromosome 17p deletion (del[17p]) is associated with poor prognosis in patients with chronic lymphocytic leukemia (CLL). Venetoclax is approved for treatment of previously untreated and relapsed/refractory (R/R) CLL, including patients with del(17p), based on the open-label, multicenter, phase 2 M13-982 trial (NCT01889186). Here, we detail the 6-year follow-up analysis for M13-982. A total of 158 patients with previously untreated (n = 5) or R/R (n = 153) del(17p) CLL received 400 mg venetoclax daily after initial ramp-up until progressive disease. After a median follow-up of 70 months, the best objective response rate (ORR) was 77% (21% complete remission [CR] and 49% partial remission [PR]), with a median duration of response (DOR) of 39.3 months (95% confidence interval [CI], 31.1-50.5). The median progression-free survival (PFS) was 28.2 months (95% CI, 23.4-37.6), and median overall survival (OS) was 62.5 months (95% CI, 51.7-not reached), with 16% of patients remaining on treatment after 6 years. Multivariable analysis did not identify statistically significant correlation between patient subgroups defined by clinical or laboratory variables and ORR or PFS. The most common grade ≥3 adverse events were neutropenia (42%), infections (33%), anemia (16%), and thrombocytopenia (16%). Post hoc comparative analyses of PFS and OS from treatment initiation, from a 24-month landmark, and by minimal residual disease status were performed between patients with del(17p) in the M13-982 and MURANO studies in the interest of understanding these data in another context. These long-term data show the continued benefits of venetoclax in patients with del(17p) CLL. The trial was registered at www.clinicaltrials.gov as #NCT01889186.

Activation of mitogen-activated protein kinase signaling and development of papillary thyroid carcinoma in thyroid-stimulating hormone receptor D633H knockin mice.

Objective: Nonautoimmune hyperthyroidism (NAH) is rare and occurs due to a constitutively activating thyroid stimulating hormone receptor (TSHR) mutation. In contrast to other thyroid nodules, no further evaluation for malignancy is recommended for hot thyroid nodules. In the first model for NAH in mice nearly all homozygous mice had developed papillary thyroid cancer by 12 months of age. Methods: To further evaluate these mice, whole exome sequencing and phosphoproteome analysis were employed in a further generation of mice to identify any other mutations potentially responsible and to identify the pathways involved in thyroid carcinoma development. Results: Only three genes (Nrg1, Rrs1, Rasal2) were mutated in all mice examined, none of which were known primary drivers of papillary thyroid cancer development. Wild-type and homozygous TSHR D633H knockin mice showed distinct phosphoproteome profiles with an enrichment of altered phosphosites found in ERK/mitogen-activated protein kinase (MAPK) signaling. Most importantly, phosphosites with known downstream effects included BRAF p.S766, which forms an inhibitory site: a decrease of phosphorylation at this site suggests an increase in MEK/ERK pathway activation. The decreased phosphorylation at BRAF p.S766 would suggest decreased AMP-activated protein kinase (AMPK) signaling, which is supported by the decreased phosphorylation of STIM1 p.S257, a downstream AMPK target. Conclusion: The modified phosphoproteome profile of the homozygous mice in combination with human literature suggests a potential signaling pathway from constitutive TSHR signaling and cAMP activation to the activation of ERK/MAPK signaling. This is the first time that a specific mechanism has been identified for a possible involvement of TSH signaling in thyroid carcinoma development.

Activation of mitogen-activated protein kinase signaling and development of papillary thyroid carcinoma in thyroid-stimulating hormone receptor D633H knockin mice.

Objective: Nonautoimmune hyperthyroidism (NAH) is rare and occurs due to a constitutively activating thyroid stimulating hormone receptor (TSHR) mutation. In contrast to other thyroid nodules, no further evaluation for malignancy is recommended for hot thyroid nodules. In the first model for NAH in mice nearly all homozygous mice had developed papillary thyroid cancer by 12 months of age. Methods: To further evaluate these mice, whole exome sequencing and phosphoproteome analysis were employed in a further generation of mice to identify any other mutations potentially responsible and to identify the pathways involved in thyroid carcinoma development. Results: Only three genes (Nrg1, Rrs1, Rasal2) were mutated in all mice examined, none of which were known primary drivers of papillary thyroid cancer development. Wild-type and homozygous TSHR D633H knockin mice showed distinct phosphoproteome profiles with an enrichment of altered phosphosites found in ERK/mitogen-activated protein kinase (MAPK) signaling. Most importantly, phosphosites with known downstream effects included BRAF p.S766, which forms an inhibitory site: a decrease of phosphorylation at this site suggests an increase in MEK/ERK pathway activation. The decreased phosphorylation at BRAF p.S766 would suggest decreased AMP-activated protein kinase (AMPK) signaling, which is supported by the decreased phosphorylation of STIM1 p.S257, a downstream AMPK target. Conclusion: The modified phosphoproteome profile of the homozygous mice in combination with human literature suggests a potential signaling pathway from constitutive TSHR signaling and cAMP activation to the activation of ERK/MAPK signaling. This is the first time that a specific mechanism has been identified for a possible involvement of TSH signaling in thyroid carcinoma development.

An inverse agonist of orphan receptor GPR61 acts by a G protein-competitive allosteric mechanism.

GPR61 is an orphan GPCR related to biogenic amine receptors. Its association with phenotypes relating to appetite makes it of interest as a druggable target to treat disorders of metabolism and body weight, such as obesity and cachexia. To date, the lack of structural information or a known biological ligand or tool compound has hindered comprehensive efforts to study GPR61 structure and function. Here, we report a structural characterization of GPR61, in both its active-like complex with heterotrimeric G protein and in its inactive state. Moreover, we report the discovery of a potent and selective small-molecule inverse agonist against GPR61 and structural elucidation of its allosteric binding site and mode of action. These findings offer mechanistic insights into an orphan GPCR while providing both a structural framework and tool compound to support further studies of GPR61 function and modulation.

Left Ventricular Ejection Fraction in Patients With Ovarian Cancer Treated With Avelumab, Pegylated Liposomal Doxorubicin, or Both.

In the phase III JAVELIN Ovarian 200 trial, 566 patients with platinum-resistant/refractory ovarian cancer were randomized 1:1:1 to receive avelumab alone, avelumab plus pegylated liposomal doxorubicin (PLD), or PLD alone. Cardiac monitoring was included for all patients. We report left ventricular ejection fraction (LVEF) data from the trial. Grade ≥3 cardiac adverse events (AEs) occurred in 4 (2.1%), 1 (0.5%), and 0 patients in the avelumab, combination, and PLD arms, respectively. LVEF decreases of ≥10% to below institutional lower limit of normal at any time during treatment were observed in 1 (0.8%), 3 (1.9%), and 2 (1.5%) patients, respectively; 4 had subsequent assessments, and these showed transient decreases. No patient had a cardiovascular AE related to LVEF decrease. This analysis is, to our knowledge, the first analysis of LVEF in patients receiving immune checkpoint inhibitors. CLINICALTRIALS.GOV IDENTIFIER: NCT02580058.

Paddle Lead Spinal Cord Stimulation in the Morbidly Obese Patient Population: Outcomes and Complications in a Single-Surgeon Cohort.

BACKGROUND: Paddle lead spinal cord stimulation (SCS) is used to treat refractory chronic pain. Morbidly obese patients seek SCS to reduce chronic pain. However, these patients face worse surgical outcomes, and the SCS literature has not evaluated safety and efficacy in this patient population. This study is the largest single-surgeon case series to date on morbidly obese patients with paddle lead SCS implantations. The primary objective is to report postoperative complication rates in morbidly obese patients receiving SCS implants. The secondary objective is to report patient-reported pain scores and Patient-Reported Outcomes Measurement Information System (PROMIS) pain interference and physical function scores in these patients. METHODS: A retrospective chart review was conducted. The patient charts were reviewed from the day of procedure consent to 6 months postop. Demographic information, pain scores, PROMIS scores, neurological complications, infections, and wound complications were documented. RESULTS: Sixty-seven patients were included. The mean preoperative BMI was 44.47 ± 4.02 kg/m2. The mean age was 58.9 ± 11.4 years old. There were no neurological complications. 3/67 (4%) developed culture-positive infections. Nine out of sixty-seven (13%) patients developed superficial wound dehiscence without underlying infection. The mean postop PROMIS physical function score was 31.6 ± 6.2 (n = 16) and the mean post-op PROMIS pain interference score was 64.0 ± 6.4 (n = 16). There was a reduction in pain scores, from 7.9 ± 1.7 preop to 5.7 ± 2.5 postop (n = 22, P = 0.004). CONCLUSIONS: Paddle lead SCS implantation is safe for morbidly obese patients. The only minimal-risk complications present were postoperative infections and wound dehiscence. Surgical care can be modified to further reduce the rates of infection and dehiscence.

The ubiquitin ligase HERC4 suppresses MafA transcriptional activity triggered by GSK3ß in myeloma by atypical K63-linked polyubiquitination.

MafA and c-Maf are close members of the Maf transcription factor family and indicators of poor prognosis of multiple myeloma (MM). Our previous study finds that the ubiquitin ligase HERC4 induces c-Maf degradation but stabilizes MafA, and the mechanism is elusive. In the present study, we find that HERC4 interacts with MafA and mediates its K63-linked polyubiquitination at K33. Moreover, HERC4 inhibits MafA phosphorylation and its transcriptional activity triggered by glycogen synthase kinase 3ß (GSK3ß). The K33R MafA variant prevents HERC4 from inhibiting MafA phosphorylation and increases MafA transcriptional activity. Further analyses reveal that MafA can also activate the STAT3 signaling, but it is suppressed by HERC4. Lastly, we demonstrate that lithium chloride, a GSK3ß inhibitor, can upregulate HERC4 and synergizes dexamethasone, a typical anti-MM drug, in inhibiting MM cell proliferation and xenograft growth in nude mice. These findings thus highlight a novel regulation of MafA oncogenic activity in MM and provide the rationale by targeting HERC4/GSK3ß/MafA for the treatment of MM.

Spinal cord stimulation: A novel approach to pain management in Dercum's disease.

Background: Dercum's Disease (DD) is a rare chronic pain syndrome in which patients experience extreme burning pain associated with subcutaneous lipomatous tissue deposits. These patients may also present with; weakness, psychiatric symptoms, metabolic derangements, sleep disturbance, impaired memory, and easy bruising. Common risk factors for DD include: obesity, Caucasian race, and female sex. The etiology of DD remains under debate while it has proven highly resistant to treatment (i.e., requiring high doses of opioids for adequate pain management). Case Description: A 48-year-old female with DD and a prior spinal cord stimulator (SCS) placed for chronic back pain, presented with recurrent back pain, and increased falling. Surgery to replace her SCS resulted in improvement in her back pain and a decreased incidence of falls. Furthermore, she noticed significant improvement in the burning pain attributed to her subcutaneous nodules; this most markedly occurred at and below the level of stimulator placement. Conclusion: A 48-year-old female with the extremely rare condition, DD experienced dramatic reduction in her pain following the successful revision of her SCS.

Adaptation of an evidence-based parenting intervention for integration into maternal-child home-visiting programs: Challenges and solutions.

Background: The objective of this study was to assess potential challenges, prioritize adaptations, and develop an implementation and research approach to integrate and study a parenting intervention for mothers in recovery from substance use disorders in community-based home-visiting programs. Method: An explanatory mixed-methods design, guided by process mapping with Failure Modes and Effects Analysis tools, and an Advisory Panel of 15 community members, identified potential implementation challenges and recommended solutions for the proposed intervention within five pre-specified domains. Thematic content analysis identified themes from detailed field notes. Results: The Advisory Panel identified 44 potential challenges across all domains. They determined that the recruitment domain was most likely to create challenges. Regarding the potential challenges, two cross-domain themes emerged: (1) development of mistrust in the community and (2) difficulty initiating and sustaining engagement. Potential solutions and adaptations to protocols are reported. Conclusion: Mistrust in the community was cited as a potentially important challenge for the delivery and study of an evidence-based parenting intervention for mothers in recovery through home-visiting programs. Adaptations to research protocols and intervention delivery strategies are needed to prioritize the psychological safety of families, particularly for groups that have been historically stigmatized.

Training home visitors in mentalization-based practice: A qualitative case study of clinical supervision in mothering from the inside out.

Attachment-based interventions are important for improving parent-child outcomes. These interventions must be scaled and made available to under-resourced communities. An important part of scaling these interventions is delineating and reproducing high-quality training, including clinical training which often requires the completion of a supervised case. However, descriptions and guidelines for clinical training are frequently broad or not available in the literature. A detailed description of clinical training could lead to further research to improve the effectiveness and dissemination of evidence-based interventions. Mothering from the Inside Out (MIO) is an attachment-based parenting intervention effective at reducing substance use and depression, improving caregiving, and enhancing child attachment. It is now being brought from research to community settings. This paper outlines the didactic and clinical training components of MIO. We then present a qualitative case study of one community-based counselor participating in the clinical training of MIO and employ qualitative methods to describe the main themes that arose during the training. We aim to illustrate how the trainer assisted the counselor in implementing the core components of MIO, which included (a) refining the language used in MIO sessions, (b) making space to explore mental states, and (c) addressing trauma. We conclude by presenting the implications of these findings.

Las intervenciones con base en la afectividad son importantes para mejorar los resultados de relación progenitor-niño. Estas intervenciones deben ser adaptadas y estar disponibles para comunidades sin recursos suficientes. Una parte importante de la adaptación de estas intervenciones es delinear y reproducir el entrenamiento de alta calidad, incluyendo entrenamiento clínico que a menudo requiere completar un caso supervisado. Sin embargo, las descripciones y los parámetros de guía para entrenamiento clínico son frecuentemente generales o no están disponibles en el material escrito. Una detallada descripción del entrenamiento clínico pudiera llevar a una posterior investigación para mejorar la eficacia y diseminación de intervenciones con base en la afectividad. Cuidados Maternales de Adentro Hacia Afuera (MIO) es una intervención de crianza con base en la afectividad que es eficaz para reducir el uso de sustancias y la depresión, mejorar la prestación de cuidado y fortalecer la afectividad del niño. Ahora se le lleva de la investigación a los escenarios comunitarios. Este artículo subraya los componentes de didáctica y entrenamiento clínico de MIO. Presentamos entonces un caso de estudio cualitativo de un consejero con base en la comunidad que participó en el entrenamiento clínico de MIO y empleamos métodos cualitativos para describir los temas principales que surgieron durante el entrenamiento. Nos propusimos ilustrar cómo el entrenador ayudó al consejero a implementar los componentes centrales de MIO, los cuales incluyen (a) refinar el lenguaje usado en la terapia, (b) abrir un espacio para explorar estados mentales, y (c) ocuparse del trauma. Concluimos con la presentación de las implicaciones que conllevan estos resultados.

Les interventions basées sur l'attachement sont importantes quand il s'agit d'améliorer les résultats parent-enfant. Ces interventions doivent être mises à l'échelle et doivent être disponibles pour toutes les communautés ayant peu de moyens. Un côté important de la mise à l'échelle de ces interventions consiste à délinéer et à reproduire une formation de haute qualité, y compris une formation clinique qui souvent exige la réalisation d'un cas supervisé. Cependant les descriptions et les lignes directrices de la formation clinique sont fréquemment larges ou ne sont pas disponibles dans des publications. Une description détaillée de formation Clinique pourrait conduire à des recherches approfondies sur la manière d'améliorer l'efficacité et la dissémination d'interventions fondées sur des données probantes. Le maternage de l'intérieur (abrégé ici MIO pour reprendre l'anglais Mothering from the Inside Out) est une intervention de parentage basée sur l'attachement qui est efficace pour ce qui concerne la réduction de toxicomanie et de dépression, l'amélioration des soins ainsi que de l'attachement de l'enfant. On le fait en ce moment passer des recherches aux contextes communautaires. Cet article décrit les composantes de formation didactique et clinique du MIO. Nous présentons ensuite une étude de cas qualitative d'un thérapeute communautaire participant à une formation clinique du MOI et employons des méthodes qualitatives pour décrire les thèmes principaux qui sont apparus durant la formation. Nous nous donnons pour but d'illustrer la manière dont le formateur a aidé le thérapeute à mettre en place les composantes essentielles du MIO, y compris (a) l'affinage du langage utilisé en thérapie, (b) la nécessité de faire de la place afin d'explorer les états mentaux, et (c) la nécessité d'aborder le trauma. Nous concluons en présentant les implications de ces résultats.

Rare case of extracranial chordoid meningioma adjacent to the carotid sheath: illustrative case.

BACKGROUND: Chordoid meningioma is a rare World Health Organization (WHO) grade 2 variant of meningioma with histological features resembling those of a chordoma. This tumor type is known for having an aggressive clinical course with a propensity for local recurrence. Most cases occur within the cranium, more specifically around the cerebral convexities. Although extracranial meningiomas of various subtypes have been documented, extracranial meningioma with a chordoid subtype is an extremely rare entity. OBSERVATIONS: The authors herein report the case of a 51-year-old female who presented with a chief complaint of dysphagia and was found to have a neck mass abutting the carotid sheath. The patient underwent resection and final pathology results revealed a WHO grade 2 chordoid meningioma. LESSONS: This case report demonstrates an atypical case of an extracranial chordoid meningioma adjacent to the carotid sheath. To the authors' knowledge, this is the first reported case of a chordoid meningioma occurring within the soft tissue of the neck.

Mitochondrial Aconitase ACO2 Links Iron Homeostasis with Tumorigenicity in Non-Small Cell Lung Cancer.

The ability of a patient tumor to engraft an immunodeficient mouse is the strongest known independent indicator of poor prognosis in early-stage non-small cell lung cancer (NSCLC). Analysis of primary NSCLC proteomes revealed low-level expression of mitochondrial aconitase (ACO2) in the more aggressive, engrafting tumors. Knockdown of ACO2 protein expression transformed immortalized lung epithelial cells, whereas upregulation of ACO2 in transformed NSCLC cells inhibited cell proliferation in vitro and tumor growth in vivo. High level ACO2 increased iron response element binding protein 1 (IRP1) and the intracellular labile iron pool. Impaired cellular proliferation associated with high level ACO2 was reversed by treatment of cells with an iron chelator, whereas increased cell proliferation associated with low level ACO2 was suppressed by treatment of cells with iron. Expression of CDGSH iron-sulfur (FeS) domain-containing protein 1 [CISD1; also known as mitoNEET (mNT)] was modulated by ACO2 expression level and inhibition of mNT by RNA interference or by treatment of cells with pioglitazone also increased iron and cell death. Hence, ACO2 is identified as a regulator of iron homeostasis and mNT is implicated as a target in aggressive NSCLC. IMPLICATIONS: FeS cluster-associated proteins including ACO2, mNT (encoded by CISD1), and IRP1 (encoded by ACO1) are part of an "ACO2-Iron Axis" that regulates iron homeostasis and is a determinant of a particularly aggressive subset of NSCLC.

Local Anesthetic Efficacy in Marijuana Users and Nonusers: A Pilot Study.

OBJECTIVE: Despite the common clinical impression that patients with a history of drug use are challenging to anesthetize with local anesthesia, literature on this clinical phenomenon is sparse. The objective of this pilot study was to assess if differences in local anesthetic efficacy for dental treatment exist between marijuana users and nonusers. METHODS: Subjects were healthy adult males and females who qualified as either chronic marijuana users or nonusers. All subjects had an asymptomatic, vital maxillary lateral incisor that responded to an electric pulp test (EPT). A standard maxillary infiltration injection technique was employed using 1.7 mL 2% lidocaine with 1:100,000 epinephrine over the test tooth, and the tooth was tested with an EPT at 3-minute intervals. RESULTS: A total of 88% of nonusers (15/17) and 61% of users (11/18) were successfully anesthetized, defined as anesthesia onset within 10 minutes and lasting at least 15 minutes. The difference in the proportion of anesthetized subjects was not statistically significant (P = .073). For subjects with successful anesthesia, there was no significant difference between nonusers and users in the onset or duration of anesthesia. CONCLUSION: No significant differences in local anesthetic efficacy with respect to local anesthetic success, onset, or duration of action were found between chronic marijuana users and nonusers. However, larger studies are likely needed to provide more definitive evidence.

Lumbar pedicle screw pseudoarthrosis salvage technique with moldable, bioabsorbable, calcium phosphate-based putty: illustrative case.

BACKGROUND: Pseudoarthrosis is a complication of spinal fixation. Risk factors include infection, larger constructs, significant medical comorbidities, and diabetes. The authors present a case report of dilated pedicle screw pseudoarthrosis salvaged with moldable, settable calcium phosphate-based putty. OBSERVATIONS: The patient presented with back pain and radiculopathy in the setting of poorly controlled diabetes. He was taken to the operating room for laminectomy and fusion complicated by postoperative infection requiring incision and drainage. He returned to the clinic 6 months later with pseudoarthrosis of the L4 screws and adjacent segment degeneration. He was taken for revision with extension of fusion. The L4 tracts were significantly dilated. A moldable, bioabsorbable polymer-based putty containing calcium phosphate was used to augment the dilated tract after decortication back to bleeding bone, allowing good purchase of screws. The patient did well postoperatively. LESSONS: There are several salvage options for clinically significant pseudoarthrosis after spinal fixation, including anterior or lateral constructs, extension, and revision of fusion. The authors were able to obtain good screw purchase with dilated screw tracts after addition of moldable, bioabsorbable polymer-based putty containing calcium phosphate. It appears that this may represent an effective salvage strategy for dilated pseudoarthropathy in select settings to support extension of fusion.

Oncohistone interactome profiling uncovers contrasting oncogenic mechanisms and identifies potential therapeutic targets in high grade glioma.

Histone H3 mutations at amino acids 27 (H3K27M) and 34 (H3G34R) are recurrent drivers of pediatric-type high-grade glioma (pHGG). H3K27M mutations lead to global disruption of H3K27me3 through dominant negative PRC2 inhibition, while H3G34R mutations lead to local losses of H3K36me3 through inhibition of SETD2. However, their broader oncogenic mechanisms remain unclear. We characterized the H3.1K27M, H3.3K27M and H3.3G34R interactomes, finding that H3K27M is associated with epigenetic and transcription factor changes; in contrast H3G34R removes a break on cryptic transcription, limits DNA methyltransferase access, and alters mitochondrial metabolism. All 3 mutants had altered interactions with DNA repair proteins and H3K9 methyltransferases. H3K9me3 was reduced in H3K27M-containing nucleosomes, and cis-H3K9 methylation was required for H3K27M to exert its effect on global H3K27me3. H3K9 methyltransferase inhibition was lethal to H3.1K27M, H3.3K27M and H3.3G34R pHGG cells, underscoring the importance of H3K9 methylation for oncohistone-mutant gliomas and suggesting it as an attractive therapeutic target.

The Wales Endoscope: The First American Cystoscope.

INTRODUCTION: Philipp Bozzini, a German army surgeon, in 1807 invented the Lichtleiter, the predecessor of the modern cystoscope. By the mid-1800s, several new instruments were created including one, a variation on Bozzini's instrument by Antoine Desormeaux in Paris. The William P. Didusch Museum of Urologic History acquired the Wales endoscope, a rare and unique cystoscope that was invented around the same time in the United States. METHODS: We researched the life of Philip Wales and the description of his cystoscope as well as Horatio Kern, the instrument maker that produced Wales' instrument. We examined the Wales cystoscope acquired by the William P. Didusch Museum. RESULTS: Philip Skinner Wales (1837-1906) was a surgeon who entered the United States Navy in 1856 and served throughout the Civil War. He organized and held charge of the Naval Hospital at New Orleans during the operations of Admiral Farragut's fleet in the Mississippi River. He was one of the first surgeons to attend President Garfield when he was shot. He was Surgeon General of the Navy (1879-1884) and founded the Museum of Naval Hygiene in Washington D.C. which later, combined with the naval laboratory and Department of Instruction, became the prototype of the Naval Medical School. In 1868 he published a series of papers in the Philadelphia Medical and Surgical Reporter on "Instrumental Diagnosis," with a paper entitled "Description of a New Endoscope." The instrument contained a metal shaft with an acute beak and used an ophthalmologic mirror to reflect light down the channel. The surgeon peered through the center hole to look into the bladder. Wales used his instrument multiple times in his private practice. Wales writes that the advantages of his cystoscope were that it was simple to produce and cheap compared to Desormeaux's endoscope. Furthermore it was light, weighing approximately 2 pounds. The main drawbacks of Wales' cystoscope were the inadequate illumination, as the light source was external and projected from the outside through a narrow channel into the bladder, and that without an optical system the image appeared relatively small. Horatio Kern, a well-known instrument maker in Philadelphia, that also supplied surgical sets and instruments for the U.S. Army during the Civil War, produced Wales' cystoscope. While he was Chief of the Bureau of Medicine, a subordinate embezzled Navy funds and Dr, Wales was court-martialed. Though he was eventually exonerated, he lived the rest of his life in disgrace in France. CONCLUSION: The Wales endoscope is unique in that it had an American inventor, was simple in design and cheap to produce. It is an important historical artifact and is one of the earliest and rarest cystoscopes developed.

Engineered SH2 Domains for Targeted Phosphoproteomics.

A comprehensive analysis of the phosphoproteome is essential for understanding molecular mechanisms of human diseases. However, current tools used to enrich phosphotyrosine (pTyr) are limited in their applicability and scope. Here, we engineered new superbinder Src-Homology 2 (SH2) domains that enrich diverse sets of pTyr-peptides. We used phage display to select a Fes-SH2 domain variant (superFes; sFes1) with high affinity for pTyr and solved its structure bound to a pTyr-peptide. We performed systematic structure-function analyses of the superbinding mechanisms of sFes1 and superSrc-SH2 (sSrc1), another SH2 superbinder. We grafted the superbinder motifs from sFes1 and sSrc1 into 17 additional SH2 domains and confirmed increased binding affinity for specific pTyr-peptides. Using mass spectrometry (MS), we demonstrated that SH2 superbinders have distinct specificity profiles and superior capabilities to enrich pTyr-peptides. Finally, using combinations of SH2 superbinders as affinity purification (AP) tools we showed that unique subsets of pTyr-peptides can be enriched with unparalleled depth and coverage.