Clonotypic VDJ rearrangements in Mixed Phenotype Acute Leukemia can be Successfully Utilized to Track Minimal Residual Disease (MRD).

INTRODUCTION: Multiplex polymerase chain reaction (PCR) and next-generation sequencing (NGS) can both be used to identify a neoplastic clonotype by targeting CDR3 and assessing rearrangements in IgH, IgK, IgL, TCR-ß, and TCR-gamma loci. The clonotypic sequence can be robustly used to track minimal residual disease (MRD). The ability to track MRD by NGS in mixed phenotype acute leukemia (MPAL) is unknown and warrants investigation. METHODS: Institutional Review Board (IRB) approval was obtained. Central Moffitt Cancer Center (MCC) database was searched to locate any patients with MPAL from over 600,000 entries. Patient charts were manually curated to identify those with clonoSEQ data, and clinical data was procured from the electronic medical record (EMR). RESULTS: Twenty-nine patients with MPAL were identified. Only 2 patients with clonoSEQ testing were found. Both demonstrated a B/myeloid phenotype, and both were bilineal. NGS (clonoSEQ) identified 4 dominant (IGH) (patient A; 8/2019) and 2 dominant sequences (patient B; 10/2019), respectively. In both patients, clonoSEQ testing successfully tracked minimal residual disease and mirrored clinical disease burden. CONCLUSIONS: This report is the first to confirm the utility of NGS-based MRD tracking in patients with MPAL and shows increased sensitivity of NGS over MRD flow cytometry.

Fit-for-purpose LC-MS/MS quantification of leukotoxin and leukotoxin diol in mouse plasma without sample pre-concentration.

LC/MS quantification of leukotoxin (LTX) and leukotoxin diol (LTXdiol) in plasma has been previously reported, however large sample volumes are required for achieving stated assay Lower Limit of Quantification (LLOQ). Reported here is a fit-for-purpose LC/MS method that reduces plasma volume from 700 to 25 µL and omits pre-concentration steps. These improvements make for a method with increased utility in mouse studies offering limited sample volumes. Additionally, omitting pre-concentration steps streamlines sample processing, which can now be completed in under 10 min. This method can be used to quickly answer if the ratio of LTX to LTXdiol changes with the dose of the therapeutic drug so this could be used as a potential biomarker for correlating PK/PD effects. No extensive assay characterization was performed before application to an exploratory in-life study. Basal levels of LTX and LTXdiol in plasma were quantified by LC-MRM across 10 individual mice, and the average signal-to-noise was 36 for LTX and 3039 for LTXdiol, with CVs of 29.4% and 15.2%, respectively. Addition of LTX and LTXdiol reference standard at 5, 25, and 75 ng/mL into pooled mouse plasma was quantifiable within 30% relative error using a surrogate matrix calibration curve ranging from 0.8 to 200 ng/mL. The average ratio of LTX to LTXdiol across the 10 mice was 0.32, consistent with previous reports. Finally, the method was applied to a mouse PK/PD study to monitor LTX/LTXdiol kinetics after a single oral dose of a soluble epoxide hydrolase inhibitor. The mean plasma ratio of LTX to LTXdiol increased up to 10-fold by 3 h post-dose followed by a decrease to near pre-dose levels by 24 h, consistent with transient inhibition of sEH-mediated conversion of LTX to LTXdiol. The method improvements described here will make subsequent quantification of LTX and LTXdiol in mouse studies significantly easier.

Application of a geographic information system-based fuzzy analytic hierarchy process model to locate potential municipal solid waste incineration plant sites: A case study of Izmir Metropolitan Municipality.

Locating candidate sites for municipal solid waste management facilities is a multi-criteria decision and involves spatial consideration that all municipalities have to deal with. This study aims to develop a holistic model for locating the most suitable facility sites in municipal solid waste management considering environmental and economic factors, and its application to an incineration facility in Izmir as a case study. A fuzzy analytic hierarchy process model was developed by integrating multi-criteria decision analysis methods and a geographic information system. A stepwise methodology was conducted, including generating a spatial database, exclusion analysis, preference analysis to determine spatial membership degrees and weights for each preference factor and generating the final land suitability map. A high-resolution land suitability map and the point vector format data of potential incineration plant sites were created as the model outputs. The case study results demonstrated that an incineration facility with a total capacity of 117 MWe could be established for energy recovery from 5649 tonnes day-1 municipal solid waste. Four locations were determined as potential incineration facility sites in the case study area. The model can be utilized for different study areas to aid decision-makers in the process of site selection for not only incineration facilities but also for other municipal solid waste management facilities.

Total Synthesis of Meayamycin B.

Meayamycin B is currently the most potent modulator of the splicing factor 3b subunit 1 and used by dozens of research groups. However, current supply for this natural product analogue is limited because of the lengthy synthetic scheme. Here, we report a more concise, more cost-effective, and greener synthesis of this compound by developing and employing a novel asymmetric reduction of a prochiral enone to afford an allylic alcohol with high enantioselectivity. In addition to this reaction, this synthesis highlights a scalable Mukaiyama aldol reaction, Nicolaou-type epoxide opening reaction, stereoselective Corey-Chaykovsky-type reaction, and a modified Horner-Wadsworth-Emmons Z-selective olefination. We also discuss a Z-E isomerization during the α,ß-unsaturated amide formation. The new synthesis of meayamycin B consists of 11 steps in the longest linear sequence and 24 total steps.

Covalent binders in drug discovery.

Covalent modulation of protein function can have multiple utilities including therapeutics, and probes to interrogate biology. While this field is still viewed with scepticism due to the potential for (idiosyncratic) toxicities, significant strides have been made in terms of understanding how to tune electrophilicity to selectively target specific residues. Progress has also been made in harnessing the potential of covalent binders to uncover novel biology and to provide an enhanced utility as payloads for Antibody Drug Conjugates. This perspective covers the tenets and applications of covalent binders.

HSP90 Inhibitor-SN-38 Conjugate Strategy for Targeted Delivery of Topoisomerase I Inhibitor to Tumors.

The clinical benefits of chemotherapy are commonly offset by insufficient drug exposures, narrow safety margins, and/or systemic toxicities. Over recent decades, a number of conjugate-based targeting approaches designed to overcome these limitations have been explored. Here, we report on an innovative strategy that utilizes HSP90 inhibitor-drug conjugates (HDC) for directed tumor targeting of chemotherapeutic agents. STA-12-8666 is an HDC that comprises an HSP90 inhibitor fused to SN-38, the active metabolite of irinotecan. Mechanistic analyses in vitro established that high-affinity HSP90 binding conferred by the inhibitor backbone could be exploited for conjugate accumulation within tumor cells. In vivo modeling showed that the HSP90 inhibitor moiety was required for selective retention of STA-12-8666, and this enrichment promoted extended release of active SN-38 within the tumor compartment. Indeed, controlled intratumoral payload release by STA-12-8666 contributed to a broad therapeutic window, sustained biomarker activity, and remarkable degree of efficacy and durability of response in multiple cell line and patient-derived xenograft models. Overall, STA-12-8666 has been developed as a unique HDC agent that employs a distinct mechanism of targeted drug delivery to achieve potent and sustained antitumor effects. These findings identify STA-12-8666 as a promising new candidate for evaluation as novel anticancer therapeutic.

Fabricating Complete Dentures with CAD/CAM and RP Technologies.

Two techological approaches for fabricating dentures; computer-aided design and computer-aided manufacturing (CAD/CAM) and rapid prototyping (RP), are combined with the conventional techniques of impression and jaw relation recording to determine their feasibility and applicability. Maxillary and mandibular edentulous jaw models were produced using silicone molds. After obtaining a gypsum working model, acrylic bases were crafted, and occlusal rims for each model were fabricated with previously determined standard vertical and centric relationships. The maxillary and mandibular relationships were recorded with guides. The occlusal rims were then scanned with a digital scanner. The alignment of the maxillary and mandibular teeth was verified. The teeth in each arch were fabricated in one piece, or set, either by CAM or RP. Conventional waxing and flasking was then performed for both methods. These techniques obviate a practitioner's need for technicians during design and provide the patient with an opportunity to participate in esthetic design with the dentist. In addition, CAD/CAM and RP reduce chair time; however, the materials and techniques need further improvements. Both CAD/CAM and RP techniques seem promising for reducing chair time and allowing the patient to participate in esthetics design. Furthermore, the one-set aligned artificial tooth design may increase the acrylic's durability.

Is it possible to diagnose caries under fixed partial dentures with cone beam computed tomography?

OBJECTIVE: The goal of this study was to determine the possibility of detecting/diagnosing caries under fixed partial dentures using cone beam computed tomography. STUDY DESIGN: A range of teeth with grade 3 to 6 caries, according to International Caries Detection and Assessment System criteria, were selected. All teeth were prepared, and three different specimens- zirconia full-ceramic, lithium disilicate full-ceramic, and metal-supported ceramic crowns-were built for each tooth. Each specimen underwent scanning by cone beam computed tomography. Opacity values were recorded and evaluated using two-way analysis of variance. The Tukey test was performed for material and slice differences, and the t test for mean gray value differences, between caries and noncaries regions of each material. RESULTS: Significant differences were detected with respect to horizontal location (anterior/posterior), restoration type (crown/bridge), material (zirconia/lithium disilicate/metal), and situation (caries/noncaries) (P < .001). Mean gray values of caries and noncaries regions were found to differ for each material. There were no significant differences with respect to vertical position. CONCLUSIONS: Cone beam computed tomography can be used as a posttreatment diagnostic technique for detecting caries under high-atomic-number fixed plate dentures. By combining high resolution and accuracy, cone beam computed tomography could provide the means for diagnosing caries without removing fixed plate dentures.

Is there a relationship between the presence of ponticulus posticus and elongated styloid process?

This study aimed to determine if there is a possible relationship concerning the presence of ponticulus posticus (PP) in patients with elongated styloid process (ESP) on three-dimensional cone beam computed tomography (CBCT) images. The presence or absence of the PP (whether partial or complete) was determined and noted as a positive or negative finding. Then, the patients with PP were evaluated for ESP. For this purpose, 3-D CBCT scanning digital images of 542 patients having ESP (247 males and 295 females) were examined retrospectively. There was a significant relationship between the presence of PP and ESP (P=.03). The results of this study suggest that there is a significant correlation between the presence of PP and ESP. To our knowledge, this is the first study investigating the relationship between the two disorders in the literature.

Evaluation of technical quality and periapical health of root-filled teeth by using cone-beam CT

Objective This study aimed to assess the quality of root fillings, coronal restorations, complications of all root-filled teeth and their association with apical periodontitis (AP) detected by cone-beam computed tomography (CBCT) images from an adult Turkish subpopulation. Material and Methods The sample for this study consisted of 242 patients (aging from 15 to 72 years) with 522 endodontically treated teeth that were assessed for technical quality of the root canal filling and periapical status of the teeth. Additionally, the apical status of each root-filled tooth was assessed according to the gender, dental arch, tooth type and age classification, undetected canals, instrument fracture, root fracture, apical resorption, apical lesion, furcation lesion and type and quality of the coronal structure. Statistical analysis was performed using percentages and chi-square test. Results The success rate of the root canal treatment was of 54.4%. The success rates of adequate and inadequate root canal treatment were not significantly different (p>0.05). Apical periodontitis was found in 228 (45.6%) teeth treated for root canals. Higher prevalence of AP was found in patients aging from 20 to 29 years [64 (27%) teeth] and in anterior (canines and incisors) teeth [97 (41%) teeth]. Conclusions The technical quality of root canal filling performed by dental practitioners in a Turkish subpopulation was consistent with a high prevalence of AP. The probable reasons for this failure are multifactorial, and there may be a need for improved undergraduate education and postgraduate courses to improve the clinical skills of dental practitioners in endodontics. .

Posttreatment diagnosis of caries under fixed restorations: a pilot study.

STATEMENT OF PROBLEM: Conventional radiographic techniques are insufficient for detecting caries under fixed restorations or for acquiring clear images of the disease. Advanced radiographic techniques such as cone beam computerized tomography may be a solution. PURPOSE: The purpose of this study was to assess the viability of detecting and characterizing caries under complete-ceramic and metal-supported crowns by using cone beam computerized tomography. MATERIAL AND METHODS: A grade 6 carious molar tooth according to the International Caries Detection and Assessment System criteria was selected. The tooth was prepared, and 3 different crowns were fabricated in a dental laboratory: yttria tetragonal zirconia polycrystal (Y-TZP) ceramic, lithium disilicate ceramic, and metal ceramic. The crowns were placed on the prepared tooth, and each specimen was subjected to cone beam computerized tomography. Opacity values were recorded and evaluated with 2-way ANOVA. Multivariate comparisons (Tukey was performed to assess the mean gray value differences of the materials between caries and dentin, and the Mann-Whitney U test was performed to assess the mean gray value differences between caries and dentin for each material (α=.05). RESULTS: Carious borders and cavities were detected under the 3 ceramic crown specimens with a cone beam computerized tomography radiographic technique. The statistical analysis of the radiopacity mean gray value was at the borderline of significance for the specimen radiographies. Significant differences were found with respect to materials, location, and the interaction between these 2 factors (P<.001). CONCLUSIONS: Cone beam computerized tomography can be used as a posttreatment diagnostic technique for detecting caries under high atomic numbered crowns and partial fixed dental prostheses.

A cone-beam computed tomography study of the root canal morphology of anterior teeth in a Turkish population.

OBJECTIVE: The aim of this study was to determine the root and canal morphology of the maxillary and mandibular anterior teeth in a Turkish population by analyzing cone-beam computed tomography (CBCT) images. MATERIALS AND METHODS: CBCT images of 417 females and 410 males with a mean age of ranging from 14 to 70 years were examined in this study. A total of 1453 maxillary central incisors, 1504 maxillary lateral incisors, 1523 maxillary canines, 1582 mandibular central incisors, 1603 mandibular lateral incisors, and 1604 mandibular canines were analyzed. The number of root canals and the canal configurations were investigated and then were classified according to Vertucci's classification. The data were analyzed with the Pearson Chi-square test. RESULTS: The Type 1 canal configuration was most prevalent in the maxillary anterior teeth (96.8-99.7%). The Types 2, 3, 4, and 5 canal configurations were observed within the range of 0-1.3%, 0-0.8%, 0-0.7%, and 0-1.8%, respectively. In the mandibular anterior teeth, the Type 1 canal configuration was most prevalent (77-95%). The Types 2, 3, 4, and 5 canal configurations were observed within a range of 0.2-2.7%, 0.2-1.6%, 0.9-5.9%, and 1.8-14.4%, respectively. The prevalence of two canals was higher in males than in females both in the maxillary and mandibular anterior teeth (P < 0.05). CONCLUSIONS: Type 1 was the most prevalent canal configuration of maxillary and mandibular anterior teeth in the Turkish population. Type 5 was the most frequently observed canal configuration of the two canalled teeth. The incidence of root canal numbers and configurations differed with sex.

Total Synthesis and Biological Studies of TMC-205 and Analogues as Anticancer Agents and Activators of SV40 Promoter.

TMC-205 is a natural fungal metabolite with antiproliferative activity against cancer cell lines. The light- and air-sensitivity prevented in-depth exploitation of this novel indole derivative. Herein, we report the first synthesis of TMC-205. On the basis of its reactivity with reactive oxygen species, we developed air-stable analogues of TMC-205. These analogues are 2-8-fold more cytotoxic than TMC-205 against HCT-116 colon cancer cell line. Importantly, at noncytotoxic dose levels, these analogues activated the transcription of luciferase reporter gene driven by simian virus 40 promoter (SV40). Further, these small molecules also inhibit firefly luciferase, presumably by direct interaction.

Evaluation of the root and canal morphology of mandibular permanent molars in a south-eastern Turkish population using cone-beam computed tomography.

OBJECTIVE: The aim of this retrospective study was to determine the root and canal morphology of the mandibular first and second permanent molars in a Turkish population using cone-beam computed tomography (CBCT). MATERIALS AND METHODS: CBCT images of mandibular first (n = 966) and second molar (n = 1165) teeth from 850 Turkish patients were evaluated. The root canal configurations were classified according to the method of Vertucci. The data were analyzed by Pearson's Chi-square test. RESULTS: The majority of mandibular molars were two rooted with three canals; however, three roots were identified in 0.05% of the first molars and 0.01% of the second molars, and 100% of the additional root canals were of type I configuration. Mesial roots had more complex canal systems with more than one canal, whereas most distal roots had a type I configuration. CONCLUSIONS: Within the limitations of this study, it can be concluded that CBCT scanning provides supplemental information about the root canal configurations of mandibular molars in a Turkish population. This study may help clinicians in the root canal treatment of mandibular molars.

Evaluation of the relationship between the maxillary posterior teeth and the sinus floor using cone-beam computed tomography.

AIM: The aim of this study was to evaluate the relationship between each root of maxillary premolars and molars and the maxillary sinus floor according to sex, sinus position, and age by decade in a Turkish population by using cone-beam computed tomography (CBCT) scanning. METHODOLOGY: We evaluated a database of 5,166 (2,680 maxillary premolars and 2,486 maxillary molars) CBCT scans obtained from 849 patients. The vertical relationship between each root of the molar and premolar teeth to the sinus floor was classified into three types: type 1, the roots penetrated into the sinus floor; type 2, the roots contacted the sinus floor; and type 3, the roots extended below the sinus floor. RESULTS: The results of the classification of each root in relationship to the sinus floor were as follows: type 3 occurred most frequently in the first (92.4 %) and second (71.6 %) premolar teeth, type 1 (34.2 %) occurred most frequently in the palatinal roots of the first molar teeth, type 3 occurred most frequently in the mesiobuccal (39.9 %) and distobuccal (39.7 %) roots of the first molar teeth, and type 2 (36.7 %) occurred most frequently in the mesiobuccal roots of the second molar teeth. No significant differences were found between the left and right sides, but several differences were found between males and females. The relationship between the posterior teeth and the sinus floor differed according to the age decade interval (p < 0.05). CONCLUSIONS: The maxillary first premolars have no relationship with the maxillary sinus floor, but the maxillary second molars are closer to the sinus floor. Also the second decade and males were most susceptible to undesirable results.

A cone-beam computed tomography study of root canal morphology of maxillary and mandibular premolars in a Turkish population.

AIM: The aim of this study was to evaluate the root canal number and configuration of maxillary and mandibular premolar teeth by gender, intervals for decades, tooth position and unilateral or bilateral occurrence in a Turkish population using Cone Beam Computed Tomography(CBCT) scanning. METHODOLOGY: CBCT images of 5496 maxillary and mandibular premolars from 849 patients were evaluated. The following was evaluated in all the images: numbers of roots and root canals, the morphology of the root canal configuration according to the Vertucci classification, male-female differences in the tooth position and male-female differences in unilateral or bilateral occurrence. The reliability data were analyzed with a chi-square test. RESULTS: The most prevalent root canal frequency was the two canals (86.2%) and type IV (76.9%) configuration for maxillary first premolar, one canal (59.7%) and type I (54.5%) canal configuration for second premolar. The incidence of one canal was higher in females and the occurrence of two or three canals was more common in males. The incidence of one canal was higher on the left side of maxillary premolars and the incidence of two canals was higher on the right side. Most mandibular first (93.5%) and second (98.5%) premolars had one canal. In general, females had one root canal of the mandibular premolar, whereas males had two or three canals. The type I configuration was most common and the incidence was higher on the right side. There were some differences found in the frequency distribution of the number of root canals and configuration of maxillary and mandibular premolar teeth according to intervals for decades. CONCLUSION: CBCT scanning provides comprehensive information about the root canal morphology of maxillary and mandibular premolar teeth. These data may help clinicians in root canal treatment of premolar teeth.

Evaluation of technical quality and periapical health of root-filled teeth by using cone-beam CT.

OBJECTIVE: This study aimed to assess the quality of root fillings, coronal restorations, complications of all root-filled teeth and their association with apical periodontitis (AP) detected by cone-beam computed tomography (CBCT) images from an adult Turkish subpopulation. MATERIAL AND METHODS: The sample for this study consisted of 242 patients (aging from 15 to 72 years) with 522 endodontically treated teeth that were assessed for technical quality of the root canal filling and periapical status of the teeth. Additionally, the apical status of each root-filled tooth was assessed according to the gender, dental arch, tooth type and age classification, undetected canals, instrument fracture, root fracture, apical resorption, apical lesion, furcation lesion and type and quality of the coronal structure. Statistical analysis was performed using percentages and chi-square test. RESULTS: The success rate of the root canal treatment was of 54.4%. The success rates of adequate and inadequate root canal treatment were not significantly different (p>0.05). Apical periodontitis was found in 228 (45.6%) teeth treated for root canals. Higher prevalence of AP was found in patients aging from 20 to 29 years [64 (27%) teeth] and in anterior (canines and incisors) teeth [97 (41%) teeth]. CONCLUSIONS: The technical quality of root canal filling performed by dental practitioners in a Turkish subpopulation was consistent with a high prevalence of AP. The probable reasons for this failure are multifactorial, and there may be a need for improved undergraduate education and postgraduate courses to improve the clinical skills of dental practitioners in endodontics.

Ruthenium-Catalyzed Olefin Metathesis after Tetra-n-butylammonium Fluoride-Mediated Desilylation.

One-pot procedures expedite organic synthesis but pose challenges in that many reagents must be compatible with each other. We discovered that the presence of (n)Bu(4)NF hindered rutheniumcatalyzed olefin metathesis when (n)Bu(4)NF-mediated desilylation and olefin metathesis were performed in one pot. This problem could be solved by the addition of (TMS)(2)O to remove fluoride anions in order to facilitate the ruthenium-catalyzed olefin metathesis.

Incidentally detected asymptomatic metal foreign bodies (two nails) in shoemaker's maxillary sinus.

The diagnosis metal foreign bodies of maxillary sinus cannot be usually made in an acute setting and patients may present with chronic symptoms due to drainage and chronic irritation of the mucosa years later. Metal foreign bodies can be generally displaced to the maxillary sinus at the time of dental intervention. In this article, we report a very interesting case of 23-year-old male shoemaker with two metal foreign bodies (nails) which were non-related with any surgical intervention or trauma and found incidentally in the right maxillary sinus.

SF3B1 haploinsufficiency leads to formation of ring sideroblasts in myelodysplastic syndromes.

Whole exome/genome sequencing has been fundamental in the identification of somatic mutations in the spliceosome machinery in myelodysplastic syndromes (MDSs) and other hematologic disorders. SF3B1, splicing factor 3b subunit 1 is mutated in 60%-80% of refractory anemia with ring sideroblasts (RARS) and RARS associated with thrombocytosis (RARS-T), 2 distinct subtypes of MDS and MDS/myeloproliferative neoplasms (MDSs/MPNs). An idiosyncratic feature of RARS/RARS-T is the presence of abnormal sideroblasts characterized by iron overload in the mitochondria, called RS. Based on the high frequency of mutations of SF3B1 in RARS/RARS-T, we investigated the consequences of SF3B1 alterations. Ultrastructurally, SF3B1 mutants showed altered iron distribution characterized by coarse iron deposits compared with wild-type RARS patients by transmission electron microscopy. SF3B1 knockdown experiments in K562 cells resulted in down-regulation of U2-type intron-splicing by RT-PCR. RNA-sequencing analysis of SF3B1 mutants showed differentially used genes relevant in MDS pathogenesis, such as ASXL1, CBL, EZH, and RUNX families. A SF3B pharmacologic inhibitor, meayamycin, induced the formation of RS in healthy BM cells. Further, BM aspirates of Sf3b1 heterozygous knockout mice showed RS by Prussian blue. In conclusion, we report the first experimental evidence of the association between SF3B1 and RS phenotype. Our data suggest that SF3B1 haploinsufficiency leads to RS formation.