Correlated libration in liquid water.

The libration spectrum of liquid H2O is resolved into an octupolar twisting libration band at 485 cm-1 and dipolar rocking-wagging libration bands at 707 and 743 cm-1 using polarization analysis of the hyper-Raman scattering (HRS) spectrum. Dipole interactions and orientation correlation over distances less than 2 nm account for the 36 cm-1 splitting of the longitudinal and transverse polarized bands of the dipolar rocking-wagging libration mode, while the intensity difference observed for the bands is the result of libration correlation over distances larger than 200 nm. The coupled rock and wag libration in water is similar to libration modes in ice. The libration relaxation time determined from the width of the spectrum is 36-54 fs. Polarization analysis of the HRS spectrum also shows long range correlation for molecular orientation and hindered translation, bending and stretching vibrations in water.

Fast investigative lung cancer pathway and endobronchial ultrasound procedure supported by rapid on-site evaluation.

INTRODUCTION: EBUS is a well-established tool for diagnosis and staging of lung cancer in a fast track investigative pathway. However, impact of ROSE in conjunction with EBUS on reduction of time to treatment decision (TTD) for cancer patients is less well known. AIMS: Our aim was to determine TTD which was defined as the number of working days from EBUS procedure to the discussion at sector lung multidisciplinary team meeting (MDT). Moreover, concordance of ROSE with final diagnosis was evaluated. METHODS: A retrospective analysis was performed of a prospective data collection in a busy teaching hospital over a four months study period (September to December 2018). RESULTS: Data from 112 patients was analyzed. There were 61 (54%) males. Mean age was of 70 years (range 43-91). WHO performance status was 0 in 20 (23%), 1 in 57 (51%), 2 in 22 (20%) and 3 in 7 (6%) patients. In total 522 needle passes were performed from 242 sampling sites. Average working days to discuss at MDT after optimal EBUS sampling was 2.087 (range 0-13 working days). ROSE concordance with final cytological diagnosis was 98.4%. The number of needle passes per site for adequate sample and diagnosis in malignant (4.929) vs non-malignant (2.776) involvement was significantly different (p value <0.0001). There was 100% sample adequacy for preliminary diagnosis, immunohistochemistry and predictive molecular testing. CONCLUSION: ROSE supported fast-investigative pathway by reducing the time to treatment decision (TTD) making at MDT. High concordance with final cytological diagnosis makes it an effective tool to inform meaningful decision making.

Synchronous uterine and bladder cancers detected in urine and vaginal samples by cytology.

Novel diagnostics for uterine cancer are urgently needed to reduce the burden of invasive testing for the majority of healthy women with postmenopausal bleeding. We have previously shown that uterine cancer cells can be detected by cytology in urine and vaginal samples with high diagnostic accuracy. Here, we demonstrate its potential to distinguish malignant cells of different aetiologies in the same urogenital biofluid sample according to their distinctive morphology and immunoprofiles. Synchronous tumours of the urogenital tract are uncommon but can cause diagnostic confusion, delays and poor outcomes. A 79-year-old woman presented to accident and emergency with postmenopausal bleeding. Voided urine and Delphi screener-collected vaginal samples were assessed by cytology and immunocytochemistry. Two malignant cell populations with distinct morphology and immunophenotypes consistent with synchronous uterine and urothelial tumours were identified. Subsequent routine diagnostics confirmed concurrent uterine carcinosarcoma and high-grade urothelial carcinoma of the bladder. This case demonstrates that cytology and adjunctive immunocytochemistry can simultaneously identify and phenotype cancers of different aetiologies from a single urogenital biofluid sample. This can help rationalise diagnostic pathways in complex, unusual cases of dual urogenital primaries.

DEveloping Tests for Endometrial Cancer deTection (DETECT): protocol for a diagnostic accuracy study of urine and vaginal samples for the detection of endometrial cancer by cytology in women with postmenopausal bleeding.

INTRODUCTION: Postmenopausal bleeding (PMB), the red flag symptom for endometrial cancer, triggers urgent investigation by transvaginal ultrasound scan, hysteroscopy and/or endometrial biopsy. These investigations are costly, invasive and often painful or distressing for women. In a pilot study, we found that voided urine and non-invasive vaginal samples from women with endometrial cancer contain malignant cells that can be identified by cytology. The aim of the DEveloping Tests for Endometrial Cancer deTection (DETECT) Study is to determine the diagnostic test accuracy of urine and vaginal cytology for endometrial cancer detection in women with PMB. METHODS AND ANALYSIS: This is a multicentre diagnostic accuracy study of women referred to secondary care with PMB. Eligible women will be asked to provide a self-collected voided urine sample and a vaginal sample collected with a Delphi screener before routine clinical procedures. Pairs of specialist cytologists, blinded to participant cancer status, will assess and classify samples independently, with differences settled by consensus review or involving a third cytologist. Results will be compared with clinical outcomes from standard diagnostic tests. A sample size of 2000 women will have 80% power to establish a sensitivity of vaginal samples for endometrial cancer detection by cytology of ≥85%±7%, assuming 5% endometrial cancer prevalence. The primary objective is to determine the diagnostic accuracy of urogenital samples for endometrial cancer detection by cytology. Secondary objectives include the acceptability of urine and vaginal sampling to women. ETHICS AND DISSEMINATION: This study has been approved by the North West-Greater Manchester West Research Ethics Committee (16/NW/0660) and the Health Research Authority. Results will be disseminated through publication in peer-reviewed scientific journals, presentation at conferences and via charity websites. TRIAL REGISTRATION NUMBER: ISRCTN58863784.

Validation of ROS1 by immunohistochemistry against fluorescent in situ hybridisation on cytology and small biopsy samples in a large teaching hospital.

OBJECTIVE: Rearranged ROS1, present in 1%-2% of non-small cell lung cancer (NSCLC) patients, usually young, never or light smokers, is assessed by fluorescence in situ hybridization (FISH) to determine eligibility for tyrosine kinase inhibitors (TKI). Immunohistochemistry (IHC) for the protein product of ROS1 rearrangement, a cost-effective alternative, is validated on cytology and small biopsy samples. METHODS: From 1 March to 31 December 2019, cytology cell blocks and small biopsy samples from a selected cohort of NSCLC patients were concurrently tested for ROS1 gene rearrangement by Vysis 6q22 Break Apart FISH probe and IHC using Cell Signalling D4D6 antibody. Mismatch cases were tested by an RNA fusion next generation sequencing (NGS) panel. RESULTS: In a prospective population of 95 cases, 91 were negative and two were positive by both FISH and IHC. Both dual positive cases were female never smokers and benefited from TKI treatment. Another two cases were positive by FISH but negative by IHC and repeat by NGS showed one to be negative but one failed. Turnaround time for IHC was 0 to 8 days from request to authorisation, whilst that of FISH was 9 to 42 days at a cost of £51 and £159 respectively. CONCLUSION: IHC to assess for the protein product of ROS1 gene rearrangement on cytology cell blocks and small biopsy samples in a routine setting is a promising screening method to assess eligibility for TKI treatment with positive and indeterminate cases confirmed by FISH or NGS as it has good negative predictive value, faster turnaround time and is cost effective, with proven technical and clinical validation.

Diagnostic accuracy of cytology for the detection of endometrial cancer in urine and vaginal samples.

Postmenopausal bleeding triggers urgent investigation by sequential invasive tests that are avoidable for the 90-95% of women who do not have endometrial cancer. A simple, non-invasive tool that accurately identifies cancer and safely reassures healthy women could transform patient care. Here we report, in a cross-sectional diagnostic accuracy study of 103 women with known cancer and 113 with unexplained postmenopausal bleeding, that urine and vaginal cytology has a combined sensitivity of 91.7% (95% CI 85.0%, 96.1%) and specificity of 88.8% (81.2%, 94.1%) for gynecological cancer detection. Cytology identifies 91 endometrial, two fallopian tube and one cervical cancer from 103 known cancer cases. In women with unexplained postmenopausal bleeding, cytology identifies all four endometrial cancers and three others (cervical, ovarian and bladder), for a 12/107 (11.2%) false positive rate. We show proof-of-principle that endometrial cancer can be detected in urine and vaginal fluid. Prospective validation of these findings will support incorporation of this non-invasive test into clinical practice.

Long-range correlation of intra-molecular and inter-molecular vibration in liquid CCl4.

Experiments measuring the polarization dependence of hyper-Raman light scattering reveal long-range correlation of molecular vibrations in liquid CCl4. The ν3 and ν1 + ν4 intra-molecular vibrations at about 770 cm-1 are strongly polarized transverse to the scattering wavevector. Weaker transverse polarization is exhibited by the ν1, ν2, and ν4 intra-molecular vibrations and by the inter-molecular collision-induced band around 0 cm-1. The observed polarization dependence is due to the correlation of the vibrations on molecules separated by about 200 nm. The strength of the observed correlation increases with the transition dipole moment for the vibration mode and is consistent with dipole-dipole coupling.

Optical and electronic solutions for power stabilization of CO2 lasers.

High pressure-temperature conditions can be readily achieved through the laser-heated diamond anvil cell (LH-DAC). A stable laser source is required for reliable in situ measurements of the sample, as the sample is small with a thermal time constant of the order of microseconds. Here, we show that the power instabilities typical of CO2 gas lasers used in LH-DAC's are ±5% at the second timescale and ∼±50% at the microsecond timescale. We also demonstrate that the pointing instability of the laser requires either a diffuser or an integrating sphere for reliable total power measurements with small sized detectors. We present a simple solution for stabilizing the power of a CO2 gas laser on the second timescale by the direct modulation of the current across the tube and another solution that stabilizes the power to the microsecond timescale by externally modulating the CO2 laser beam. Both solutions can achieve a ±0.3% power stability.

Vibration overtone hyperpolarizability measured for H2.

The second hyperpolarizability (γ) of the H2 molecule was measured by gas-phase electric field induced second harmonic generation at the frequencies of the one-photon resonance for the 3-0 Q(J) overtone transitions (v, J = 0, J → 3, J for J = 0, 1, 2, and 3). The magnitude of the resonant contribution to γ was measured with 2% accuracy using the previously determined non-resonant γ for calibration. Pressure broadening and frequency shift for the transitions were also measured. A theoretical expression for the resonant vibrational γ contribution in terms of transition polarizabilities is compared to the observations. The measured γ resonance strength is 4%-14% larger than the results obtained from this theoretical expression evaluated using ab initio transition polarizabilities.

CT Volumetry and Basic Texture Analysis as Surrogate Markers in Advanced Non-small-cell Lung Cancer.

INTRODUCTION: We evaluated volumetric tumor measurements and computed tomography texture analysis as prognostic indicators in patients with advanced non-small-cell lung cancer when compared with the unidimensional tumor size measurements used in Response Evaluation Criteria in Solid Tumors (RECIST). PATIENTS AND METHODS: In a retrospective review, computed tomography examinations in 77 patients with advanced non-small-cell lung cancer were evaluated before and after 2 cycles of chemotherapy. Baseline and changes in tumor diameter, volume, and texture were analyzed. Survival was analyzed with Cox regression analysis and Kaplan-Meier survival statistics. RESULTS: Cox regression analysis demonstrated that only change in tumor volume (exp(B) = 1.006; P = .02) and the initial sum of the largest target lesion diameters predicted survival (exp(B) = 1.013; P = .02). Kaplan-Meier statistics demonstrated that patients with an initial sum of the largest target lesion diameters less than 88 mm had median survival time of 587 days (95% confidence interval [CI], 269-905 days), compared with the survival of those with larger tumor burden of 407 days (95% CI, 235-579 days). Patients in whom tumor volume decreased by more than 29% had a median survival time of 622 days (95% CI, 448-796 days), compared with 305 days for those with less decrease (95% CI, 34-240 days). CONCLUSION: This study demonstrates that change in lung tumor volume is a better marker of patient survival than change of unidimensional diameter measurements in our cohort. If confirmed in larger studies, this suggests that volumetry might improve clinical decision-making for individual patients and allow for faster assessment of new treatments.

Preclinical Efficacy and Safety Comparison of CD3 Bispecific and ADC Modalities Targeting BCMA for the Treatment of Multiple Myeloma.

The restricted expression pattern of B-cell maturation antigen (BCMA) makes it an ideal tumor-associated antigen (TAA) for the treatment of myeloma. BCMA has been targeted by both CD3 bispecific antibody and antibody-drug conjugate (ADC) modalities, but a true comparison of modalities has yet to be performed. Here we utilized a single BCMA antibody to develop and characterize both a CD3 bispecific and 2 ADC formats (cleavable and noncleavable) and compared activity both in vitro and in vivo with the aim of generating an optimal therapeutic. Antibody affinity, but not epitope was influential in drug activity and hence a high-affinity BCMA antibody was selected. Both the bispecific and ADCs were potent in vitro and in vivo, causing dose-dependent cell killing of myeloma cell lines and tumor regression in orthotopic myeloma xenograft models. Primary patient cells were effectively lysed by both CD3 bispecific and ADCs, with the bispecific demonstrating improved potency, maximal cell killing, and consistency across patients. Safety was evaluated in cynomolgus monkey toxicity studies and both modalities were active based on on-target elimination of B lineage cells. Distinct nonclinical toxicity profiles were seen for the bispecific and ADC modalities. When taken together, results from this comparison of BCMA CD3 bispecific and ADC modalities suggest better efficacy and an improved toxicity profile might be achieved with the bispecific modality. This led to the advancement of a bispecific candidate into phase I clinical trials.

Optimal design, anti-tumour efficacy and tolerability of anti-CXCR4 antibody drug conjugates.

Antibody-drug conjugates (ADCs) are promising therapies for haematological cancers. Historically, their therapeutic benefit is due to ADC targeting of lineage-restricted antigens. The C-X-C motif chemokine receptor 4 (CXCR4) is attractive for targeted therapy of haematological cancers, given its expression in multiple tumour types and role in cancer "homing" to bone marrow. However, CXCR4 is also expressed in haematopoietic cells and other normal tissues, raising safety challenges to the development of anti-CXCR4 ADCs for cancer treatment. Here, we designed the first anti-CXCR4 ADC with favourable therapeutic index, effective in xenografts of haematopoietic cancers resistant to standard of care and anti-CXCR4 antibodies. We screened multiple ADC configurations, by varying type of linker-payload, drug-to-antibody ratio (DAR), affinity and Fc format. The optimal ADC bears a non-cleavable linker, auristatin as payload at DAR = 4 and a low affinity antibody with effector-reduced Fc. Contrary to other drugs targeting CXCR4, anti-CXCR4 ADCs effectively eliminated cancer cells as monotherapy, while minimizing leucocytosis. The optimal ADC selectively eliminated CXCR4+ cancer cells in solid tumours, but showed limited toxicity to normal CXCR4+ tissues, sparing haematopoietic stem cells and progenitors. Our work provides proof-of-concept that through empirical ADC design, it is possible to target proteins with broad normal tissue expression.

Third harmonic scattering in liquids.

Third harmonic scattering (THS) from liquids has been observed and analyzed in several recent papers. It is considered to be analogous to second harmonic scattering (hyper-Rayleigh scattering) and to provide a means for measuring the second hyperpolarizability tensor of molecules in a liquid. However, the observed signal for a pure solvent is in fact mainly due to coherent third harmonic generation followed by Rayleigh scattering and direct incoherent THS (direct THS) makes only a small contribution (<2% for parallel polarized THS). This invalidates the internal reference method and the polarization analysis that has been applied for pure liquids. Theoretical comparison for the two processes, extensive experimental measurements for CCl4 liquid, SiO2 glass, and CCl2F2 gas and survey measurements for D2O, CDCl3, CD3CN, and (CD3)2SO liquid, are presented.

Response to "Comment on 'Water-water correlations in electrolyte solutions probed by hyper-Rayleigh scattering'" [J. Chem. Phys. 149, 167101 (2018)].

The Comment proposes that the discrepancy between two second harmonic scattering (SHS) experiments for D2O electrolyte solutions with 100 ns and 190 fs laser pulses is due to unexplained processes for 100 ns pulses that give non-quadratic power dependence for the second harmonic signal. However, the different power dependence of the second harmonic scattering signals measured with 100 ns and 190 fs laser pulses is due to changes in laser beam propagation and focal intensity caused by the combined effect of thermal defocusing and Kerr lens self-focusing. Non-quadratic power dependence does not explain the discrepancy in the second harmonic scattering results.

Simple imaging for the diamond anvil cell: Applications to hard-to-reach places.

The employment of high-pressure gases as a pressure-transmitting medium, sample, or reactant for diamond anvil cell experiments is widespread. As a pressure transmitter, high-pressure gases are crucial to forming quasi-hydrostatic compression atmospheres for samples inside the uniaxially driven cell. We describe an optical design for forming high-resolution images of the gasket and sample chamber of the diamond anvil cell under high gas pressures in a gas loading apparatus. Our design is simple, is of low-cost, and may be easily adapted to suit gas loading apparatus of any design, as well as other common hard-to-reach environments in diamond anvil cell experiments, i.e., those with large stand-off distances, such as cryostats.

Clinical utility of tumor genomic profiling in patients with high plasma circulating tumor DNA burden or metabolically active tumors.

BACKGROUND: This retrospective study was undertaken to determine if the plasma circulating tumor DNA (ctDNA) level and tumor biological features in patients with advanced solid tumors affected the detection of genomic alterations (GAs) by a plasma ctDNA assay. METHOD: Cell-free DNA (cfDNA) extracted from frozen plasma (N = 35) or fresh whole blood (N = 90) samples were subjected to a 62-gene hybrid capture-based next-generation sequencing assay FoundationACT. Concordance was analyzed for 51 matched FoundationACT and FoundationOne (tissue) cases. The maximum somatic allele frequency (MSAF) was used to estimate the amount of tumor fraction of cfDNA in each sample. The detection of GAs was correlated with the amount of cfDNA, MSAF, total tumor anatomic burden (dimensional sum), and total tumor metabolic burden (SUVmax sum) of the largest ten tumor lesions on PET/CT scans. RESULTS: FoundationACT detected GAs in 69 of 81 (85%) cases with MSAF > 0. Forty-two of 51 (82%) cases had ≥ 1 concordance GAs matched with FoundationOne, and 22 (52%) matched to the National Comprehensive Cancer Network (NCCN)-recommended molecular targets. FoundationACT also detected 8 unique molecular targets, which changed the therapy in 7 (88%) patients who did not have tumor rebiopsy or sufficient tumor DNA for genomic profiling assay. In all samples (N = 81), GAs were detected in plasma cfDNA from cancer patients with high MSAF quantity (P = 0.0006) or high tumor metabolic burden (P = 0.0006) regardless of cfDNA quantity (P = 0.2362). CONCLUSION: This study supports the utility of using plasma-based genomic assays in cancer patients with high plasma MSAF level or high tumor metabolic burden.

RN765C, a low affinity EGFR antibody drug conjugate with potent anti-tumor activity in preclinical solid tumor models.

Epidermal growth factor receptor (EGFR) is a clinically validated target and often overexpressed in some solid tumors. Both EGFR tyrosine kinase inhibitors and ligand-blocking antibodies have been approved for treatment of NSCLC, head and neck cancers and colorectal cancers. However, clinical response is limited and often accompanied by significant toxicities due to normal tissue expression. To improve the effectiveness of targeting EGFR while minimizing the toxicities on normal tissues, we developed a low-affinity anti-EGFR antibody drug conjugate (ADC), RN765C. Potent in vitro cytotoxicity of RN765C, with nanomolar to subnanomolar EC50, was observed on a panel of cancer cell lines expressing moderate to high level of EGFR. In contrast, RN765C was less effective in killing normal human keratinocytes, presumably due to its lower receptor expression. Mechanistically, RN765C has multiple modes of action: inducing payload mediated mitotic arrest and cell death, blocking EGFR pathway signal and mediating antibody dependent cell cytotoxicity. In preclinical studies, a single dose of RN765C at 1.5-3 mg/kg was generally sufficient to induce tumor regression in multiple cell line and patient-derived xenograft models, including those that are resistant to EGFR-directed tyrosine kinase inhibitors. Our data support further investigation of RN765C in the clinic to treat EGFR expressing solid tumors.

First-in-human randomized clinical trials of the safety and efficacy of tanezumab for treatment of chronic knee osteoarthritis pain or acute bunionectomy pain.

INTRODUCTION: The neurotrophin nerve growth factor has a demonstrated role in pain transduction and pathophysiology. OBJECTIVES: Two randomized, double-blind, placebo-controlled, phase 1 studies were conducted to evaluate safety, tolerability, and analgesic efficacy of single doses of tanezumab, a humanized anti-nerve growth factor monoclonal antibody, in chronic or acute pain. METHODS: In the first study (CL001), patients with moderate to severe pain from osteoarthritis (OA) of the knee received a single intravenous infusion of tanezumab (3-1000 µg/kg) or placebo in a dose-escalation (part 1; N = 42) or parallel-arm (part 2; N = 79) study design. The second study (CL002) was a placebo-controlled dose-escalation (tanezumab 10-1000 µg/kg; N = 50) study in patients undergoing bunionectomy surgery. RESULTS: Adverse event rates were generally similar across treatments. Most adverse events were generally mild to moderate in severity and no patients discontinued as a result of adverse events. Adverse events of abnormal peripheral sensation were more common with higher doses of tanezumab (≥100 µg/kg) than with placebo. These were generally mild to moderate in severity. Tanezumab provided up to 12 weeks of effective analgesia for OA knee pain, with statistically significant improvements at doses ≥100 µg/kg (P < 0.05). By contrast, no trend for analgesic activity was found when tanezumab was administered 8 to 16 hours before bunionectomy. CONCLUSIONS: The demonstration of a favorable safety profile and clinical efficacy in OA pain supports clinical development of tanezumab as a potential treatment for chronic pain conditions.

Effects of Monoclonal Antibodies against Nerve Growth Factor on Healthy Bone and Joint Tissues in Mice, Rats, and Monkeys: Histopathologic, Biomarker, and Microcomputed Tomographic Assessments.

Tanezumab, an anti-nerve growth factor (NGF) antibody, is in development for management of chronic pain. During clinical trials of anti-NGF antibodies, some patients reported unexpected adverse events requiring total joint replacements, resulting in a partial clinical hold on all NGF inhibitors. Three nonclinical toxicology studies were conducted to evaluate the effects of tanezumab or the murine precursor muMab911 on selected bone and joint endpoints and biomarkers in cynomolgus monkeys, Sprague-Dawley rats, and C57BL/6 mice. Joint and bone endpoints included histology, immunohistochemistry, microcomputed tomography (mCT) imaging, and serum biomarkers of bone physiology. Responses of bone endpoints to tanezumab were evaluated in monkeys at 4 to 30 mg/kg/week for 26 weeks and in rats at 0.2 to 10 mg/kg twice weekly for 28 days. The effects of muMab911 at 10 mg/kg/week for 12 weeks on selected bone endpoints were determined in mice. Tanezumab and muMab911 had no adverse effects on any bone or joint parameter. There were no test article-related effects on bone or joint histology, immunohistochemistry, or structure. Reversible, higher osteocalcin concentrations occurred only in the rat study. No deleterious effects were observed in joints or bones in monkeys, rats, or mice administered high doses of tanezumab or muMab911.