Novel methodology comparing two assay formats for the assessment of immunogenicity from clinical trial samples

Aim: We present a novel methodology to compare results between distinct immunogenicity assays, performed by two laboratories, for the same biotherapeutic. Materials & methods: Human serum pools from clinical trials were generated to provide representative immunogenicity titers. Pools were evaluated at two laboratories in a blinded fashion to assess the effect of assay format and laboratory change on clinical interpretation of immunogenicity results. Results: The laboratories validated two different assay formats and demonstrated comparable sensitivity and drug tolerance. Overall, the comparisons in assay format and laboratory ensured a comparable ability to detect treatment-emergent antidrug antibodies for a biotherapeutic. Conclusion: We have established an approach, using pooling of patient samples, that allows for the interlaboratory comparisons without creating duplicative results.

Prospective cohort study of voice outcomes following secondary tracheoesophageal puncture in gastric pull-up reconstruction after total laryngopharyngoesophagectomy

Background Gastric pull-up is a reconstructive option for circumferential defects after resection of advanced laryngopharyngeal malignancy. Voice loss is expected and vocal rehabilitation remains a challenge. Our study objectives were to investigate the feasibility of secondary tracheoesophageal puncture following gastric pull-up and to analyze voice outcomes. Methods This was a prospective cohort study of patients with advanced laryngopharyngeal malignancies who underwent gastric pull-up and secondary tracheoesophageal puncture between 1988 and 2017 at a tertiary-care academic institution. Objective acoustic measures included fundamental frequency and vocal intensity. Perceptual analysis was performed using voice recordings (“Rainbow Passage”) randomly presented in a blinded fashion to four clinicians using the validated GRBAS scale. Speech intelligibility was assessed in a blinded fashion using a validated 7-point scale. Additionally, the Voice Handicap Index-10 was administered as a validated patient self-reporting tool. Results Ten patients (7 male, 3 female) were included, all of whom preferentially used tracheoesophageal puncture for communication. These patients had abnormal median fundamental frequency of 250 (interquartile range (IQR) 214–265) Hz and a limited median vocal intensity of 65.8 (IQR 64.1–68.3) dB. Perceptual analysis (GRBAS) revealed a median ‘moderate’ degree of impairment [grade 2 (IQR 2–3), roughness 2 (IQR 2–3), breathiness 3 (IQR 2–3), asthenia 2 (IQR 1–2), strain 2 (IQR 1–2)] as did median intelligibility scores [median 5 (IQR 4–7)]. Most patients self-reported an abnormal voice handicap-10 [median 26.5 (IQR 22.8–35.0)]. Conclusion Secondary tracheoesophageal puncture is a safe and feasible option for voice rehabilitation after gastric pull-up. Although analyses demonstrated moderate subjective and objective impairment, tracheoesophageal puncture provided patients with a self-reported means of functional verbal communication and was their preferred method of communication. Graphical abstract

OR19-02 Luteinizing Hormone/Human Chorionic Gonadotropin Receptor Protein Expression in Adrenocortical Progenitor Cells, Aldosterone Producing Cell Clusters and Adrenal Adenomas Derived from Postmenopausal Women

Objective/Background Adrenal pathologies are more common in women than men. Embryologically the adrenals and gonads develop from the adrenogenital ridge with differential migration and differentiation. We hypothesized that in adult females there are adrenocortical progenitor cells that express the LH/hCG-R and proliferate in response to elevated LH. Indeed, several case reports demonstrated LH/hCG-R expression in adrenal secretory tumors in postmenopausal and pregnant females. In aging adults, nests of cells known as aldosterone-producing cell clusters (APCCs) that may be precursors to aldosterone producing adenomas are frequently detected. We retrospectively studied the immunohistochemical expression of LH/hCG-R in normal adrenals, adrenal adenomas and APCCs in archival specimens derived from post-menopausal women. Methods Archival specimens from adrenal adenomas derived from 23 women >55 years of age were examined. Clinical data was obtained in a blinded fashion and hormonal data was available in 9/23 cases; 6/9 were secreting cortisol and 3/9 adenomas were secreting aldosterone. In addition, 6 samples derived from a repository of normal adrenal tissues from deceased kidney donors (1 male, and 5 postmenopausal females) were studied. All specimens were immunostained for LH/hCG-R and the adrenal stem cell marker DLK1 that facilitates the maintenance of an undifferentiated phenotype. The normal adrenal tissues were also stained for aldosterone synthase (CYP11B2) to detect APCCs. The slides were reviewed and graded by a pathologist in a blinded fashion. Results Expression of LH/hCG-R was demonstrated in both normal and adenomatous tissues in all 23 specimens. The staining in adenomas was heterogeneous, with clusters of densely stained LH/hCG-R positive cells in all specimens. There were less densely stained clusters in normal adjacent adrenocortical tissue that was most prominent in the subcapsular, zona glomerulosa region, an area where the putative adrenal cortical stem cells are found as well as the zona reticularis. Double staining for the stem cell marker DLK1 and LH/hCG-R confirmed that these cells represent adrenocortical progenitor cells. CYP 11B2 immunohistochemistry of normal adrenals demonstrated cell foci dipping from the capsule into the zona fasciculata classified as APCCs that co-expressed cytoplasmic LH/hCGR. Conclusion Adrenal adenomas and APCCs derived from postmenopausal women exhibited heterogeneous but strong immunohistochemical expression of LH/hCG-R in all samples. Interestingly, DLK1-positive adrenocortical stem cells in the subcapsular zone also expressed LH/hCG-R. These data may provide insights into the female predominance of adrenal pathologies, particularly in postmenopausal women with high LH levels. The LH/hCG-R may be a viable target for treatment of adrenal adenomas in postmenopausal women.

Intraoperative Tilted Posteroanterior View for the Measurement of Distal Radius Articular Step-Off

The purpose of this study is to more accurately determine distal radius articular step-off in the posteroanterior (PA) view. A cadaveric forearm was osteotomized with varying amounts of articular displacement. A second osteotomy was made through the distal radius metaphysis to create four positions of tilt in the lateral plane (5° and 15° dorsal tilt; 5° and 15° volar tilt). Using fluoroscopy, the beam was positioned in the lateral plane from 25° volar to 20° dorsal, separated by 5° increments, obtaining modified PA images of the distal radius in its various configurations. The images were randomly evaluated for step-off by three hand surgeons in a blinded fashion. Statistical analysis was performed to determine the accuracy between estimated and actual step-off and was demonstrated to be greater when the PA view was parallel to the distal radius tilt in the lateral plane, for all four configurations of distal radius tilt. Data pertaining to the distal radius with 0 mm of step-off did not demonstrate the PA view, parallel to the distal radius tilt, to be superior than the PA views not parallel to the tilt; reaffirming that with anatomic reduction, any fluoroscopic image exhibits good alignment. This study confirms that the most accurate method of accessing PA step-off is to first determine the tilt of the radius on a lateral film and then align the beam in the PA plane to match this tilt.

Abstract WP305: Uniting Hospital Leaders in a Competitive Region to Improve Intravenous Thrombolysis Rates: Analysis of the SETRAC Stroke Registry

Uniting Hospital Leaders in a Competitive Region To Improve Intravenous Thrombolysis Treatment Rates: Analysis of the SETRAC Stroke Registry Background: The Southeast Texas Regional Advisory Council (SETRAC) convenes representatives from area hospitals belonging to different health systems to discuss how to improve the delivery of stroke care in a nine county area that includes Houston, Texas. In 2014, area hospitals agreed to focus upon improving tissue plasminogen activator (tPA) administration rates in the area. Hypothesis: By sharing stroke metrics in a blinded fashion on a consistent basis, tPA administration rates will improve. The theory was tested by tracking the following metrics: a. percent of ischemic stroke patients receiving tPA. b. percent of treated patients receiving tPA within 60 minutes of arrival at the hospital. Methods: SETRAC aligned regional metrics with stroke treatment and timing metrics in Get With The Guidelines®-Stroke beginning in Q3 2014. SETRAC compiled and shared this data with stroke coordinators and hospital administrators on a quarterly basis. Bar graphs were utilized for benchmarking hospitals in a blinded fashion. Linear tests were performed to validate increases in the percent of patients treated with tPA and the percent of patients treated within 60 minutes of arrival at the hospital. Results from hospitals that consistently reported data for each of the six quarters were included in the analysis. Results: Twenty-six area hospitals reported 10,396 ischemic stroke patients between Q3 2014 through Q4 2015. In that time, tPA administration increased from 10.3% in Q3 2014 to 12.0% in Q4 2015 (p<0.001). Further, tPA administration within 60 minutes of arrival in emergency room increased from 55.3% in Q3 2014 to 66.7% in Q4 2015 (p<0.001). In conclusion, following the uniting of hospital leaders and utilizing nationally aligned stroke metrics, a higher percentage of ischemic stroke patients received tPA. Of those that received tPA, an improvement in the timeliness of administration was also realized.

Changes in Nitric Oxide Releases of the Contralateral Acupoint during and after Laser Acupuncture at Bilateral Same-Name Acupoints in Human

Objective.The purpose of the study was to examine the effects of laser acupuncture (LA) at right Neiguan (RPC6)/left Neiguan (LPC6) acupoints on the releases of nitric oxide (NO) in the treated and contralateral/nontreated PC6, compared to the nonacupoint control area.Methods. 24 mW LA at RPC6, LPC6, and nonacupoint in 22 healthy subjects for 40 min: sterilized dialysis tube was taped to the nontreated PC6/nonacupoint during the treatment and immediately taped to the treated and nontreated PC6/nonacupoint after LA removal. NO-scavenging compound was injected into the tube for 40 min to absorb the molecular which was tested by spectrophotometry in a blinded fashion.Results. LA-induced NO releases over PC6 acupoints for the nontreated and treated sides all significantly increased after LA removal, but for the nontreated acupoints they did not change during LA stimulation. LA at RPC6 induced the more release of the NO at contralateral side than stimulating LPC6, but not on nonacupoints. The results suggest that LA-induced NO release over contralateral acupoint and NO release resulting from the lateralized specificity all are different and specific to the acupoint within different time course.Conclusions. LA-evoked NO release over acupoints could improve the neurogenic, endothelial activity of the vessel wall to further facilitate microcirculation.

Response of Local Nitric Oxide Release to Manual Acupuncture and Electrical Heat in Humans: Effects of Reinforcement Methods

This study was to examine the influences of manual acupuncture (MA) and electrical heat corresponding to reinforcing methods on nitric oxide (NO) release over the skin regions in humans. A device with collecting solution was taped to the skin surface along pericardium (PC) or lung (LU) meridian. Acupuncture needles were gently inserted into PC 4 with reinforcing stimulation (low force/rate) for 20 minutes in the MA group. LU11 on the finger was heated (43-44°C) by electrical heat for 20 minutes. Biocapture was consecutively conducted for two 20-minute intervals during and after each treatment. Total nitrite and nitrate (NOx-) in the collecting samples were quantified using chemiluminescence in blinded fashion. BaselineNOx-levels are higher and tended to be higher over PC and LU acupoints during the 1st biocapture.NOx-levels over PC regions were consistently increased by MA during both intervals.NOx-concentrations over LU acupoints were increased and tended to be increased by electrical heat in the 1st and 2nd biocapture. The results suggest that reinforcing MA and electrical heat induce NO released from the local skin regions with higher levels at acupoints, which improve local circulation and contribute to the beneficial effects of the therapies.

Taste of Clindamycin and Acetaminophen

This study evaluated the taste palatability of liquid clindamycin and acetaminophen products on the market. Subjects rated the palatability of 3 clindamycin suspensions, 1 amoxicillin suspension (tasted twice), an acetaminophen elixir, and an acetaminophen suspension in a randomized blinded fashion on a 0 to 5 scale. Forty-six adults aged 20 to 82 years volunteered for this study. Means (and 95% confidence intervals) were as follows: amoxicillin-first taste 3.6 (3.3-3.9), amoxicillin-second taste 3.5 (3.2-3.7). Clindamycin Rising, Perrigo, Greenstone; 2.0 (1.6-2.5), 3.0 (2.7-3.3), and 2.2 (1.8-2.6), respectively. Acetaminophen elixir 0.6 (0.4-0.8) and acetaminophen suspension 3.4 (3.1-3.6). One clindamycin tasted significantly better than the others. Additionally, although 2 acetaminophen formulations are currently available over-the-counter, the suspension is more palatable and less costly. Medicaid drug programs that perpetuate the use of elixir should change their coverage to save money and provide patients access to better tasting acetaminophen.

Response to Intravenous Lidocaine in a Patient with SUNCT Syndrome

Short-lasting unilateral neuralgiform headache with conjunctival injection and tearing (SUNCT) syndrome represents a serious therapeutic and diagnostic challenge, since it is usually refractory to most drugs and lacks biological markers. Response to intravenous lidocaine administration has been reported in some patients while it has failed in others. We report a patient with SUNCT syndrome who showed a clear-cut and consistent response to intravenous lidocaine therapy, which proved superior to placebo in a single-blinded fashion. Intravenous lidocaine should be considered in all patients with SUNCT syndrome. Response to this therapy could represent a diagnostic tool.

Predictive Value of the Pre-Operative CT Scan for Splenectomy in Patients with Primary, and Post Essential Thrombocythemia/Post Polycythemia Vera Myelofibrosis.

BACKGROUND: Splenectomy is performed for palliating symptoms in patients with primary myelofibrosis (PMF; as well as post essential thrombocythemia myelofibrosis/ post polycythemia myelofibrosis (Mesa et. al. Cancer2006;107(2):361–70). Pre-operative computed tomograms (CTs) of the spleen are frequently performed for estimating spleen size and anatomy, but sometimes raise concern over radiographic splenic masses. We performed a retrospective analysis of splenectomized PMF patients to determine the predictive value of CT in these patients. METHODS: Splenectomized patients, with confirmed PMF (and POST ET/PV MF), with an available pre-operative CT scan were included. Pre-operative CT scans were reviewed in a blinded fashion by a radiologist (B.B.), and histopathology was concurrently reviewed in a blinded fashion by a hematopathologist (C.Y.L.), with attention for radiographic evidence of splenic masses, splenic infarction and hemorrhage, evidence of extra-splenic masses. Additionally splenic volume was calculated by CT through an algorithm using maximal splenic length, width, and slice size (prorated ellipsoid where volume = (30+.58(XxYxZ)). Results from each review were compared and analyzed against clinical outcomes and documented splenic mass. RESULTS: Thirty-eight patients had concurrent CT and splenic pathology available underwent splenectomy for myelofibrosis (20 PMF; median age at diagnosis 64.6 years (range (46–79), 52% males). Splenic volume estimations by CT correlated well with splenic mass measurements. Splenic volume estimation by CT was 2365 CC (range 736–11355 CC), compared with a measured median splenic mass of 2000 mg (range 730–11750 mg) (bivariate analysis p<0.001). Splenic lesions were identified in a total of 33 patients (87%), 10 only by histopathology (30%), 3 only by CT (9%), and 20 by both (61%). Histopathologically 15 patients had localized hemorrhage, 13 had splenic infractions, and 8 had nodular foci of extramedullary hematopoiesis. Only 2 patients had evidence of an alternative malignancy (concurrent chronic lymphocytic leukemia (CLL, n =1) or systemic mast cell disease (n = 1). CT was very sensitive for detecting focal low density lesions (LDLs), which are roughly spherical regions of decreased contrast enhancement in the spleen, and detected the majority of macroscopic lesions. The presence of splenic infarctions histologically was associated with decreased post splenectomy survival (p=0.02; median 7.2 months vs. 15.0 months), however the sensitive (but not specific) presence of LDL lesions by CT did not have prognostic significance. The CT appearance of splenic hemorrhage was nonspecific, and since these areas of parenchymal hemorrhage were likely subacute/chronic, they could not be reliably distinguished from other LDLs. CONCLUSIONS: Pre-operative CT in patients whom undergo palliative splenectomy for myelofibrosis is a sensitive tool for determining the presence of small splenic lesions (hemorrhage, infarction and EMH). Additionally splenic volume estimation by CT, correlates well with measured splenic mass and may assist in operative planning. Splenic lesions in PMF (post ET/PV MF) by CT rarely represented an alternative malignant process, and should likely not be the sole indication for splenectomy. Splenic infarctions were associated with decreased survival after splenectomy, however CT is not always able to distinguish these lesions from EMH and hemorrhage.