Real-world performance of blood-based proteomic profiling in first-line immunotherapy treatment in advanced stage non-small cell lung cancer.

PURPOSE: Immune checkpoint inhibition (ICI) therapy has improved patient outcomes in advanced non-small cell lung cancer (NSCLC), but better biomarkers are needed. A clinically validated, blood-based proteomic test, or host immune classifier (HIC), was assessed for its ability to predict ICI therapy outcomes in this real-world, prospectively designed, observational study. MATERIALS AND METHODS: The prospectively designed, observational registry study INSIGHT (Clinical Effectiveness Assessment of VeriStrat® Testing and Validation of Immunotherapy Tests in NSCLC Subjects) (NCT03289780) includes 35 US sites having enrolled over 3570 NSCLC patients at any stage and line of therapy. After enrolment and prior to therapy initiation, all patients are tested and designated HIC-Hot (HIC-H) or HIC-Cold (HIC-C). A prespecified interim analysis was performed after 1-year follow-up with the first 2000 enrolled patients. We report the overall survival (OS) of patients with advanced stage (IIIB and IV) NSCLC treated in the first-line (ICI-containing therapies n=284; all first-line therapies n=877), by treatment type and in HIC-defined subgroups. RESULTS: OS for HIC-H patients was longer than OS for HIC-C patients across treatment regimens, including ICI. For patients treated with all ICI regimens, median OS was not reached (95% CI 15.4 to undefined months) for HIC-H (n=196) vs 5.0 months (95% CI 2.9 to 6.4) for HIC-C patients (n=88); HR=0.38 (95% CI 0.27 to 0.53), p<0.0001. For ICI monotherapy, OS was 16.8 vs 2.8 months (HR=0.36 (95% CI 0.22 to 0.58), p<0.0001) and for ICI with chemotherapy OS was unreached vs 6.4 months (HR=0.41 (95% CI 0.26 to 0.67), p=0.0003). HIC results were independent of programmed death ligand 1 (PD-L1). In a subgroup with PD-L1 ≥50% and performance status 0-1, HIC stratified survival significantly for ICI monotherapy but not ICI with chemotherapy. CONCLUSION: Blood-based HIC proteomic testing provides clinically meaningful information for immunotherapy treatment decision in NSCLC independent of PD-L1. The data suggest that HIC-C patients should not be treated with ICI alone regardless of their PD-L1 expression.

Factors influencing operative time in arthroscopic rotator cuff repair: a comparison of knotless single-row vs. transosseous equivalent dual-row techniques.

BACKGROUND: Despite the widespread use of arthroscopic rotator cuff repair (aRCR), there remains considerable debate on the benefits of a dual-row vs. a single-row (SR) repair technique. This study compares operative time of a knotless SR technique with transosseous equivalent (TOE) dual-row technique for aRCR and defines patient-specific factors that affect operative time. METHODS: Data from 118 patients who underwent aRCR with a knotless SR technique was compared with data from 95 patients who underwent aRCR with a TOE technique by a single surgeon between 2014 and 2018. Baseline patient demographic information and operative time were recorded and compared between the 2 groups. Subgroup analysis was performed to determine if demographic information or tear size influenced operative time. RESULTS: The average operative time in the SR group was 75.68 minutes and the average operative time in the TOE group was 89.24 minutes (P < .001). When controlling for all concomitant procedures, the operative time in the TOE group was 8.1 minutes longer than the SR group (P = .029). Average tear size in an anterior-posterior direction was larger in the TOE group vs. the SR group, 26.09 mm vs. 15.18 mm (P < .001). CONCLUSION: When controlling for concomitant procedures, a knotless, TOE dual-row technique for aRCR adds an average of 8 minutes' operative time compared with a knotless SR technique. This was despite a significantly larger tear size in the TOE group.

Desmoplastic Fibroma in the Distal Humerus of a 14-Year-Old Boy: A Case Report.

CASE: We present the case of a 14-year-old male who presented with a pathologic elbow fracture. Imaging demonstrated a lytic distal humerus metaphyseal lesion with a soap bubble appearance. After fracture healing, frozen sections obtained at open biopsy suggested fibrous tissue; thus, curettage and bone grafting was performed. Pathology confirmed desmoplastic fibroma. The patient experienced a recurrence 18 months later and had repeat curettage and allograft strutting. He had no evidence of re-recurrence 2 years after the second surgery. CONCLUSIONS: Desmoplastic fibroma may mimic the presentation of many bone lesions. Recurrence is common despite curettage and bone grafting. This patient performed well without wide resection.

Total Knee Arthroplasty After Knee Arthroscopy in Patients Older Than 50 Years.

Several orthopedic registries have described the incidence of total knee arthroplasty (TKA) in patients who have undergone knee arthroscopy. Patient risk factors may play a role in the conversion rate from knee arthroscopy to TKA. This study quantifies the incidence of conversion of knee arthroscopy to TKA from a US mixed-payer database and describes some common patient risk factors for conversion. The medical records of more than 50 million patients who were treated between 1998 and 2014 were mined with a commercially available software platform. During the study period, a total of 68,090 patients older than 50 years underwent knee arthroscopy for partial meniscectomy, chondroplasty, or debridement. Reported rates of TKA at 1, 2, and 3 years after arthroscopy were 10.1%, 13.7%, and 15.6%, respectively. Obesity, depressive disorder, rheumatoid arthritis, diabetes, and age 70 years and older were associated with increased relative risk of conversion to TKA at 2 years. When obesity was combined individually with the top 5 other risk factors, no combination produced a higher relative risk than that of obesity alone. Patients who were 50 to 54 years of age had the lowest incidence of conversion to TKA (8.3%, P<.001). Men had a lower incidence of conversion to TKA (11.3%) than women (15.8%, P<.001). This information can help surgeons to counsel patients on the incidence of TKA after knee arthroscopy and identify preoperative risk factors that increase risk. [Orthopedics. 2016; 39(6):e1041-e1044.].

A stitch in time: an unusual cause of enterovesical fistula.

A 64-year-old woman with a history of a Stamey procedure for stress incontinence 20 years previously, underwent a diagnostic flexible cystoscopy to investigate recurrent urinary sepsis. Cystoscopic examination demonstrated a large vesicular calculus. When traction was applied to the calculus it was shown to be attached to a 10 cm length of non-absorbable suture material. The suture was attached to two further calculi, apparently originating from outside the bladder wall. All visible calculi and the suture were removed. In the following weeks the patient reported passage of air and faeculent material per-urethra. A repeat cystoscopy revealed an opening in the posterolateral bladder wall and CT confirmed the presence of an enterovesical fistula. Laparoscopy demonstrated a fistula between the terminal ileum and bladder wall. A further 10 cm length of non-absorbable suture material was removed from the terminal ileum and omentum. Laparoscopic repair of the fistula was performed successfully and the patient has remained symptom free following the procedure.

Effect of azathioprine intolerance on outcomes of inflammatory bowel disease: a cross-sectional study.

BACKGROUND: Azathioprine is well established for the maintenance of remission in patients with inflammatory bowel disease (IBD). However, a significant proportion of patients are intolerant to azathioprine. It is not clear if intolerance of azathioprine is a marker of poor prognosis for patients who will have a more aggressive disease or be more likely to require surgery. OBJECTIVE: To determine if intolerance to azathioprine is a marker of poor prognosis, indicating patients who have a more aggressive disease course, and to analyse the risk factors and causes of intolerance. METHODS: A cross-sectional study using the Milton Keynes Hospital IBD database was performed to compare azathioprine-intolerant and azathioprine-tolerant patients. RESULTS: Two hundred and thirty-nine patients met the inclusion criteria comprising 141 patients with Crohn's disease (CD) and 98 patients with ulcerative colitis (UC). Overall, 28.0% of patients were intolerant to azathioprine. Risk factors for intolerance were female sex and age 50-70 years. Common reasons for intolerance were nausea and vomiting (34.3%), deranged liver function tests (28.4%) and headaches (11.9%). In patients with UC, there was no statistical difference in the disease activity scores between those who were azathioprine intolerant versus tolerant. In patients with CD, azathioprine intolerance was associated with significantly worse disease activity; 25% fewer patients were in clinical remission and 20% more had moderate/severe disease. Rates of surgery were similar between the groups for both UC/CD. CONCLUSIONS: We conclude that azathioprine intolerance acts as a surrogate marker for patients with CD who in future have poorer symptom control. Azathioprine intolerance marks out a group of patients with CD in whom increased vigilance of symptom control and early escalation of treatment is required.

Identification and characterization of interactions between abscisic acid and mitochondrial adenine nucleotide translocators.

ABA (abscisic acid) is a plant hormone involved in important processes including development and stress responses. Recent reports have identified a number of plant ABA receptors and transporters, highlighting novel mechanisms of ABA action. In the present paper we describe application of a chemical proteomics approach leading to the identification of mitochondrial ANTs (adenine nucleotide translocators) as ABA-interacting proteins. Initial in vitro studies confirmed inhibition of ANT-dependent ATP translocation by ABA. Further analysis demonstrated ANT-dependent uptake of ABA into both recombinant Arabidopsis thaliana ANT2-containing proteoliposomes and native isolated spinach mitochondria; the latter with a Km of 3.5 µM and a Vmax of 2.5 nmol/min per g of protein. ATP was found to inhibit ANT-dependent ABA translocation. Specificity profiles highlight the possibility of mechanistic differences in translocation of ABA and ATP. Finally, ABA was shown to stimulate ATPase activity in spinach mitochondrial extracts. ABA concentrations in plant cells are estimated to reach the low micromolar range during stress responses, supporting potential physiological relevance of these in vitro findings. Overall, the present in vitro work suggests the possibility of as yet uncharacterized mechanisms of ABA action in planta related to inhibition of mitochondrial ATP translocation and functional localization of ABA in the mitochondrial matrix.

Identification and characterization of a unique cysteine residue proximal to the catalytic site of Arabidopsis thaliana carotenoid cleavage enzyme 1.

AtCCD1 and AtNCED3 are related carotenoid cleavage enzymes from Arabidopsis thaliana that catalyze the oxidative cleavage of, respectively, the 9,10 (9',10') double bonds of carotenoid substrates such as beta-carotene, and the 11,12 double bond of 9-cis epoxycarotenoids. Although the cellular and cleavage functionalities of these enzymes have been reported, their mechanisms and related structural environments mediating these disparate specificities in homologous enzymes have not been well characterized. By relating the differences observed in UV and visible light absorption and Cu(II) electron paramagnetic signals to variations in sequence alignments and 3-D homology models of the two A. thaliana enzymes, we identified a putatively proximal cysteine residue (Cys352) in AtCCD1 that is not conserved in AtNCED3. Spectral analysis of the Cys to Ala mutant confirmed its uniqueness and proximity to the metal binding site, but precluded any role for the residue in the mediation of the observed metal binding affinity or associated steric constraint differences. Further analysis of kinetic substrate cleavage properties indicated a decrease in Vmax and a subtle increase in Km for the C352A mutant compared with those observed for the wild-type, thus confirming catalytic site proximity and suggesting possible roles for the unique cysteine in the modulation of substrate affinity and (or) the reaction rate of AtCCD1.

Elephantiasis nostrum verrucosa of the abdomen: clinical results with tazarotene.

Elephantiasis nostras verrucosa represents an uncommon yet distinct clinical entity resulting from chronic lymphedema of an extremity or body region. Characterized by profound non-pitting edema with cobblestone-like papules, plaques, and nodules, it typically occurs secondary to infections, surgeries, tumor obstruction, radiation, congestive heart failure, and obesity. This progressively deformative disorder has been treated with various medical and surgical measures. In the following case report, the history, clinical, and pathologic appearance of elephantiasis nostras verrucosa are discussed, as well as newer treatment options.

A coumermycin/novobiocin-regulated gene expression system.

Regulated expression of transgenes in mammals is an important technique in both functional genomic studies and clinical applications. Here we describe a regulated gene expression system for mammals, based on coumarin-switched dimerization of the bacterial DNA gyrase B subunit (GyrB). The transactivator was constructed by fusing the GyrB activator to the bacterial lambda repressor-binding domain. The antibiotic coumermycin in nanomolar concentrations activated the transgene through binding of the homodimerized chimeric transactivator to the lambda operator located upstream of a minipromoter. More significantly, addition of novobiocin, an antagonist of coumermycin, promptly switched off expression of the gene by abolishing coumermycin-induced dimerization of the transactivator. Site-directed mutagenesis of the lambda repressor-binding domain resulted in significant reduction of basal expression levels and an induction reaching four orders of magnitude in stably transfected 293A cells in response to coumermycin. The capability of this inducible system for tightly regulated gene expression was demonstrated by the ready generation of stable cell lines inducibly expressing the proapoptotic bax gene in mammalian cells. Hence, this novel coumarin switch-on/switch-off system should broaden the utility of regulated gene expression, particularly when rapid on/off interchange is required.

Characterization of selective estrogen receptor modulator (SERM) activity in two triarylethylene oxybutyric acids.

Previously we identified 4-[1-(4-hydroxyphenyl)-2-phenylbuten-1-yl]phenoxy-n-butyric acid (4HBA) and its des-hydroxy analog (BA) as potential selective estrogen receptor modulators (SERMs) in the ovariectomized (OVX) rat. The aim of the present study was to characterize comprehensively the effects of 4HBA and BA in both the OVX rat and in estrogen-responsive cells. Thus, 4HBA was found to be an estrogen antagonist with partial agonist efficacy in estrogen-responsive reporter gene and estrogen-dependent proliferation assays (MVLN cells and MCF-7 human breast cancer cells, respectively). In the OVX rat, 4HBA and BA were equally effective and comparable to other known SERMs regarding (a) serum cholesterol reduction and suppression of serum markers of excessive bone metabolism, and (b) partial agonist efficacy in reproductive tissue relative to steroidal estrogens. Like steroidal estrogens, both compounds increased serum triglyceride levels, with BA being more effective in this regard. The maximal effects of 4HBA on all of these parameters except cholesterol lowering were seen at oral doses of 0.4 micromol/kg/day; maximal cholesterol lowering required doses of 10 micromol/kg/day. In OVX rat liver 9S fraction, BA was found to be efficiently converted to a single hydroxylated metabolite, 4HBA. These results suggest that the effects of BA in the OVX rat might, in part, be a consequence of biotransformation to 4HBA, and that those of 4HBA and BA in the OVX rat and in estrogen-responsive cells are qualitatively similar to those of SERMs such as tamoxifen and raloxifene.