An auristatin-based antibody-drug conjugate targeting EphA2 in pancreatic cancer treatment.

Pancreatic adenocarcinoma, a highly aggressive form of cancer with a poor prognosis, necessitates the development of innovative treatment strategies. Our prior research showcased the growth-inhibiting effects of the anti-EphA2 antibody drug hSD5 on pancreatic cancer tumors. This antibody targets and induces the degradation of the EphA2 receptor while also prompting the antibody's internalization. A deeper dive into the hSD5 Fab crystallographic structure and docking studies revealed that hSD5's CDRH3 drives the primary interaction between hSD5 and the EphA2 active site. In this study, we developed a novel antibody-drug conjugate (ADC)-the auristatin-based hSD5-vedotin specifically targeting EphA2 in pancreatic cancer cells. This ADC aims at the tumor-specific antigen EphA2, triggering endocytosis and releasing the conjugated payload molecule Monomethyl auristatin E (MMAE), amplifying the tumor-killing effect. Upon cellular entry, hSD5-vedotin demonstrated an impressive tumor-killing response, inhibiting tumor cell growth and promoting apoptosis even at lower antibody concentrations. In a pancreatic cancer xenograft animal model, hSD5-vedotin showcased the potential to suppress tumor growth entirely. Notably, potential immune resistance responses were also observed in recurrent pancreatic cancer tumors. Our empirical results underscore the possibility of developing hSD5-vedotin further, which we anticipate will have a broader and more potent therapeutic impact on pancreatic cancer and other EphA2-related cancers.

Spin-Enhanced Lateral Flow Immunoassay for High-Sensitivity Detection of Nonstructural Protein NS1 Serotypes of the Dengue Virus.

Dengue fever is a global mosquito-borne viral infectious disease that has, in recent years, rapidly spread to almost all regions of the world. Lack of vaccination and directed treatment makes detection at the infection's early stages extremely important for disease prevention and clinical care. In this paper, we developed a rapid and highly sensitive dengue detection tool using a novel platform of diagnosis, called spin-enhanced lateral flow immunoassay (SELFIA) with a fluorescent nanodiamond (FND) as a reporter. Taking advantage of the unique magneto-optical properties of negatively charged nitrogen-vacancy centers in the FND, the SELFIA platform utilizes alternating electromagnetic fields to modulate signals from FND's fluorescence to provide sensitive and specific results. With sandwich SELFIA, we could efficiently detect all four dengue non-structural protein (NS1) serotypes (DV1, DV2, DV3, and DV4). The lowest detection concentration of the dengue NS1 antigens varied from 0.1 to 1.3 ng/mL, which is among the lowest limits of detection to date. The FND-based SELFIA technique is up to 500 and 5000 times more sensitive than carbon black and conventional gold nanoparticles, respectively. By using different anti-NS1 antibodies, we could differentiate the NS1 antigen serotypes contained in the tested samples via three simultaneous assays. Proposed SELFIA allows for both qualitative and quantitative differentiation between different NS1 protein serotypes, which will assist in the development of a highly sensitive and specific detection platform for dengue screening that has the potential to detect the disease at its early stages, especially in high-risk and limited-resource areas.

Crystal Structure of TCPTP Unravels an Allosteric Regulatory Role of Helix α7 in Phosphatase Activity.

The T-cell protein tyrosine phosphatase (TCPTP/PTPN2) targets a broad variety of substrates across different subcellular compartments. In spite of that, the structural basis for the regulation of TCPTP's activity remains elusive. Here, we investigated whether the activity of TCPTP is regulated by a potential allosteric site in a comparable manner to its most similar PTP family member (PTP1B/PTPN1). We determined two crystal structures of TCPTP at 1.7 and 1.9 Å resolutions that include helix α7 at the TCPTP C-terminus. Helix α7 has been functionally characterized in PTP1B and was identified as its allosteric switch. However, its function is unknown in TCPTP. Here, we demonstrate that truncation or deletion of helix α7 reduced the catalytic efficiency of TCPTP by ∼4-fold. Collectively, our data supports an allosteric role of helix α7 in regulation of TCPTP's activity, similar to its function in PTP1B, and highlights that the coordination of helix α7 with the core catalytic domain is essential for the efficient catalytic function of TCPTP.

Vibrio cholerae biofilm scaffolding protein RbmA shows an intrinsic, phosphate-dependent autoproteolysis activity.

Vibrio cholerae is the causative agent of the diarrheal disease cholera, for which biofilm communities are considered to be environmental reservoirs. In endemic regions, and after algal blooms, which may result from phosphate enrichment following agricultural runoff, the bacterium is released from biofilms resulting in seasonal disease outbreaks. However, the molecular mechanism by which V. cholerae senses its environment and switches lifestyles from the biofilm-bound state to the planktonic state is largely unknown. Here, we report that the major biofilm scaffolding protein RbmA undergoes autocatalytic proteolysis via a phosphate-dependent induced proximity activation mechanism. Furthermore, we show that RbmA mutants that are defective in autoproteolysis cause V. cholerae biofilms to grow larger and mechanically stronger, correlating well with the observation that RbmA stability directly affects microbial community homeostasis and rheological properties. In conclusion, our biophysical study characterizes a novel phosphate-dependent breakdown pathway of RbmA, while microbiological data suggest a new, sensory role of this biofilm scaffolding element.

Development of a Versatile and Modular Linker for Antibody-Drug Conjugates Based on Oligonucleotide Strand Pairing.

Linker design is crucial to the success of antibody-drug conjugates (ADCs). In this work, we developed a modular linker format for attaching molecular cargos to antibodies based on strand pairing between complementary oligonucleotides. We prepared antibody-oligonucleotide conjugates (AOCs) by attaching 18-mer oligonucleotides to an anti-HER2 antibody through thiol-maleimide chemistry, a method generally applicable to any immunoglobulin with interchain disulfide bridges. The hybridization of drug-bearing complementary oligonucleotides to our AOCs was rapid, stoichiometric, and sequence-specific. AOCs loaded with cytotoxic payloads were able to selectively target HER2-overexpressing cell lines such as SK-BR-3 and N87, with in vitro potencies similar to that of the marketed ADC Kadcyla (T-DM1). Our results demonstrated the potential of utilizing AOCs as a highly versatile and modular platform, where a panel of well-characterized AOCs bearing DNA, RNA, or various nucleic acid analogs, such as peptide nucleic acids, could be easily paired with any cargo of choice for a wide range of diagnostic or therapeutic applications.

Development of a highly efficient, strongly coupled organic light-emitting diode based on intracavity pumping architecture.

We report a highly efficient polariton organic light-emitting diode (POLED) based on an intracavity pumping architecture, where an absorbing J-aggregate dye film is used to generate polariton modes and a red fluorescent OLED is used for radiative pumping of emission from the lower polariton (LP) branch. To realize the device with large-area uniformity and adjustable coupling strength, we develop a spin-coating method to achieve high-quality J-aggregate thin films with controlled thickness and absorption. From systematic studies of the devices with different J-aggregate film thicknesses and OLED injection layers, we show that the J-aggregate film and the pump OLED play separate roles in determining the coupling strength and electroluminescence efficiency, and can be simultaneously optimized under a cavity design with a good LP-OLED emission overlap for effective radiative pumping. By increasing the absorption with thick J-aggregate film and improving the electron injection of pump OLED with Li2CO3 interlayer, we demonstrate the POLED with a large Rabi splitting energy of 192 meV and a maximum external quantum efficiency of 1.2%, a record efficiency of POLEDs reported so far. This POLED architecture can be generally applied for exploration of various organic materials to realize novel polariton devices and electrically pumped lasers.

Structural basis of polyethylene glycol recognition by antibody.

BACKGROUND: Polyethylene glycol (PEG) is widely used in industry and medicine. Anti-PEG antibodies have been developed for characterizing PEGylated drugs and other applications. However, the underlying mechanism for specific PEG binding has not been elucidated. METHODS: The Fab of two cognate anti-PEG antibodies 3.3 and 2B5 were each crystallized in complex with PEG, and their structures were determined by X-ray diffraction. The PEG-Fab interactions in these two crystals were analyzed and compared with those in a PEG-containing crystal of an unrelated anti-hemagglutinin 32D6-Fab. The PEG-binding stoichiometry was examined by using analytical ultracentrifuge (AUC). RESULTS: A common PEG-binding mode to 3.3 and 2B5 is seen with an S-shaped core PEG fragment bound to two dyad-related Fab molecules. A nearby satellite binding site may accommodate parts of a longer PEG molecule. The core PEG fragment mainly interacts with the heavy-chain residues D31, W33, L102, Y103 and Y104, making extensive contacts with the aromatic side chains. At the center of each half-circle of the S-shaped PEG, a water molecule makes alternating hydrogen bonds to the ether oxygen atoms, in a similar configuration to that of a crown ether-bound lysine. Each satellite fragment is clamped between two arginine residues, R52 from the heavy chain and R29 from the light chain, and also interacts with several aromatic side chains. In contrast, the non-specifically bound PEG fragments in the 32D6-Fab crystal are located in the elbow region or at lattice contacts. The AUC data suggest that 3.3-Fab exists as a monomer in PEG-free solution but forms a dimer in the presence of PEG-550-MME, which is about the size of the S-shaped core PEG fragment. CONCLUSIONS: The differing amino acids in 3.3 and 2B5 are not involved in PEG binding but engaged in dimer formation. In particular, the light-chain residue K53 of 2B5-Fab makes significant contacts with the other Fab in a dimer, whereas the corresponding N53 of 3.3-Fab does not. This difference in the protein-protein interaction between two Fab molecules in a dimer may explain the temperature dependence of 2B5 in PEG binding, as well as its inhibition by crown ether.

Application of a cubic phase plate to a reflecting telescope for extension of depth of field.

Wavefront coding is an accepted technique to extend the depth of field in incoherent optical systems. We present the design of a reflecting telescope with wavefront coding by employing a cubic phase plate (CPP). We propose a method to reduce the CPP size by changing its position from the aperture stop to a secondary mirror without varying the point-spread function, which is insensitive to defocusing. This change in position reduces the diameter by almost three orders and increases the third-order coefficients of the xy polynomial surface by two orders, thus easing manufacturing.

Admittance analysis of broadband omnidirectional near-perfect absorber in epsilon-near-zero mode.

In this paper, we propose a broadband omnidirectional near-perfect absorber that transforms light energy into heat. In contrast to previous research on structural metamaterials, this study focuses on light absorption in the epsilon-near-zero (ENZ) layers without any structural patterns. Chromium (Cr) thin films were applied as ENZ layers. Using the admittance method, we found the proper thicknesses of SiO2 layers to match the incident medium and achieve perfect absorption. Also, the absorber is angular insensitive up to 60°. The temperature of the absorber increases from room temperature to 42°C, which is 4°C higher than the uncoated substrate at 38°C, after exposure to sunlight for 20 min.

Crystal Structure of PigA: A Prolyl Thioester-Oxidizing Enzyme in Prodigiosin Biosynthesis.

Prodigiosin is an intensely red pigment comprising three pyrroles. The biosynthetic pathway includes a two-step proline oxidation catalyzed by phosphatidylinositol N-acetylglucosaminyltransferase subunit A (PigA), with flavin adenine dinucleotide (FAD) as its cofactor. The enzyme is crystallized in the apo form and in complex with FAD and proline. As an acyl coenzyme A dehydrogenase (ACAD) family member, the protein folds into a ß-sheet flanked by two α-helical domains. PigA forms a tetramer, which is consistent with analytical ultracentrifugation results. FAD binds to PigA in a similar way to that in the other enzymes of the ACAD family. The variable conformations of loop ß4-ß5 and helix αG correlate well with the structural flexibility required for substrate entrance to the Re side of FAD. Modeling with PigG, the acyl carrier protein, suggests a reasonable mode of interaction with PigA. The structure helps to explain the proline oxidation mechanism, in which Glu244 plays a central role by abstracting the substrate protons. It also reveals a plausible pocket for oxygen binding to the Si side of FAD.

Miniaturized laser readout head for application in a double-wedge prism scanner.

Double-wedge prism scanners are typically used for one-dimensional (1D) scanning in imaging systems. This type of scanner operates on the principle of two identical wedge prisms counter-rotating around the optical axis with identical angular velocity in order to move the image formed by the optical system in the plane of detector arrays. However, the relationship between the deviation and rotation angles is not linear. The proposed solution to this problem involves using a metal ring inscribed with line markers. The interval between each of the line markers is unequal, and each line marker corresponds with the unequal angular orientation of the wedge prisms. The line markers are detected by a laser readout head (LRH), which converts the line marker signals to electric clock pulses to correct for the nonlinear distortion of the double-wedge prism scanner. This study presents a miniaturized LRH, electric driver circuit, and photodetector amplifier for this purpose.

Baseline cardiac output and its alterations during ibuprofen treatment for patent ductus arteriosus in preterm infants.

BACKGROUND: Infants with hemodynamically significant patent ductus arteriosus (PDA) may physiologically compensate with a supranormal cardiac output (CO). As such, a supranormal CO may be a surrogate marker for a significant PDA or indicate a failed response to PDA closure by ibuprofen. Electrical cardiometry (EC) is an impedance-based monitor that can continuously and non-invasively assess CO (COEC). We aimed to trend COEC through ibuprofen treatment for PDA in preterm infants. METHODS: We reviewed our database of preterm infants receiving ibuprofen for PDA closure. Response to ibuprofen was defined as no ductal flow in echocardiography ≤24 h after treatment. Responders were compared with gestational age (GA) and postnatal age matched non-responders and their trends of COEC were compared. Both groups' baseline COEC were further compared to the reference infants without PDA. RESULTS: Eighteen infants (9 responders and 9 non-responders) with median (interquatile range) GA 27.5 (26.6-28.6) weeks, birthweight 1038 (854-1218) g and age 3.5 (3.0-4.0) days were studied. There were positive correlations between COEC and ductal diameter and left atrium/ aortic root ratio (r = 0.521 and 0.374, p < 0.001, respectively). Both responders and non-responders had significantly higher baseline COEC than the reference. Although there was no significant within-subject alteration of COEC during ibuprofen treatment, there was a between-subject difference indicating non-responders had generally higher COEC. CONCLUSIONS: The changes of COEC during pharmacological closure of PDA is less drastic compared to surgical closure. Infants with PDA had higher baseline COEC compared to those without PDA, and non-responders had higher COEC especially at baseline compared to responders.

Double-wedge prism scanner for application in thermal imaging systems.

In this paper, double-wedge prisms, also known as Risley prisms, and composed of two wedge prisms, are presented with assemblies in four configurations. In the double-wedge prism scanner, the center of the two wedge prisms is aligned with the optical axis; thus, the optical system and mechanical structure are simple. In contrast to conventional scanners, its assembly is compact, robust, and insensitive to vibrations and wobbles. However, the relationship between the deviated and rotated angles is not linear, and the trajectory of the scan pattern is nonrectilinear; therefore, double-wedge prisms have rarely been used in imaging systems as a one-dimensional scanner until now. This study presents the optical characteristics of double-wedge prisms and proposes methods to solve the problems mentioned above. The experimental results demonstrate that the double-wedge prisms can be used in an imaging system as a one-dimensional scanner.

Analytic inverse solutions for Risley prisms in four different configurations for positing and tracking systems.

The conventional ray deviation formula and analytical inverse solution for Risley prisms are traditionally derived by the first-order paraxial method, which is simplified and does not provide sufficient accuracy. When a light ray enters the wedge prism, the component that is perpendicular to the base side is deviated by the wedge prism and the component parallel to the base side is uninfluenced. In this study, the problem is presented and analyzed from the scalar form of Snell's law, and 2D vector algebra is used to solve the problem. The rotation and refracting angles of a single wedge prism are obtained by analyzing the propagation through the prism of incident rays from different directions, and ray deviation formulas are derived. Four configurations are described with ray deviation formulas and analytical inverse solutions deduced from the corresponding formulas for a single wedge prism. In the final section, two examples are considered for the four configurations. The numeric results demonstrate the accuracy of the inverse solution provided by the proposed method.

Varying stress of SiOxCy thin films deposited by plasma polymerization.

SiOxCy thin films were deposited by plasma polymerization. The stress of the deposited SiOxCy thin films can be modified by adjusting the beam current, the anode voltage, and the flow rate of hexamethyldisiloxane (HMDSO) gas and oxygen. Reducing the beam current or increasing the flow rate of HMDSO gas increased the linear/cage structure ratio and turned the stress of the SiOxCy thin films from compressive to tensile. The linear/cage structure ratio can be adjusted by changing the composite parameter, W[FM]c/[FM]m, to control the stress of the deposited plasma polymer films. Multilayers of TiO2/SiO2/TiO2 were coated on a SiOxCy plasma polymer film herein, reducing their stress by 70% from 0.06 to 0.018 GPa. The refractive index is 1.55, and the absorption coefficient is less than 10-4 at 550 nm of the SiOxCy films. Superior optical performances of SiOxCy thin films make their use in optical thin films.

A single amino acid in VP2 is critical for the attachment of infectious bursal disease subviral particles to immobilized metal ions and DF-1 cells.

VP2 protein is the primary host-protective immunogen of infectious bursal disease virus (IBDV). His249 and His253 are two surface histidine residues in IBDV subviral particles (SVP), which is formed by twenty VP2 trimers when the VP2 protein of a local isolate is expressed. Here, a systemic study was performed to investigate His249 or/and His253 on self-assembly, cell attachment and immunogenicity of SVP. Point-mutagenesis of either or both histidine residues to alanine did not affect self-assembly of the SVP, but the SVP lost its Ni-NTA binding affinity when the His253 was mutated. Indirect immunofluorescence assays and inhibitory experiments also showed that His253 is essential for SVP to attach onto the DF-1 cells and to inhibit IBDV infection of DF-1 cells. Finally, enzyme-linked immunosorbent assays and chicken protection assays demonstrated that SVP with a mutation of His253 to alanine induced comparable neutralizing antibody titers in chickens as the wild-type SVP did. It was concluded that VP2's His253, a site not significant for the overall immunogenicity induced by SVP, is crucial for the binding affinity of SVP to Ni-NTA and the attachment of an IBDV host cell line. This is the first paper to decipher the role of His253 played in receptor interaction and immunogenicity.

Fabry-Perot based metal-dielectric multilayered filters and metamaterials.

The traditional three-layered metal-dielectric-metal Fabry-Perot filter is developed as a new metal-dielectric multilayered band-pass filter. Our design method allows metal and dielectric films to be alternatively arranged to achieve a narrow and high transmission peak and the peak height remains unchanged for any number of metal films arranged in the multilayer. Furthermore, the equivalent refractive index of a subwavelength metal-dielectric multilayer could be negative real at the passband of the filter and such metamaterial exhibits stronger figure of merit than a previous result. By choosing a material with high refractive index as the dielectric film, such metamaterial exhibits a pass band that depends weakly on the angle of incidence.

Reduced threshold of optically pumped amplified spontaneous emission and narrow line-width electroluminescence at cutoff wavelength from bilayer organic waveguide devices.

We present a detailed study of the optically and electrically pumped emission in the BSB-Cz/PVK bilayer waveguide devices. By optical pumping we demonstrate that PVK as a spacer between fluorescent BSB-Cz and ITO electrode allows the significant reduction of the threshold for amplified spontaneous emission (ASE) of BSB-Cz. The simulation provides a better understanding of how the PVK thickness affects the waveguide mode field distribution and hence the ASE threshold of BSB-Cz. On the other hand, the BSB-Cz/PVK bilayer OLED exhibits the external quantum efficiency of >1% and anisotropic electroluminescence with spectrally narrowed edge emission at the cutoff wavelength controlled by the BSB-Cz thickness. When tuning the cutoff wavelength to match the peak gain of BSB-Cz, we demonstrate an intense, particularly narrow edge emission (~5 nm) without obvious degradation of efficiency at a high current density of 1000 mA/cm2, suggesting a reliable device performance for high-power applications and further exploration of electrically-pumped ASE.

Measuring and modeling the inconspicuous iridescence of Formosan blue magpie's feather (Urocissacaerulea).

The iridescence of the blue feathers of the Formosan blue magpie (Urocissacaerulea) is not conspicuous when the viewing angle is less than 40°. The spongy medullary keratin inside the feather barbs is investigated by two-dimensional Fourier analysis of transmission electron microscopic images of various positions on a barb to explain this unique characteristic. The orientation of the quasi-ordered nanostructure varies depending on its position of the feather barb. The predicted reflectance increases with the distance of the nanostructures from the vertex of the feather barb, and this result agrees closely with measurements.

Biopsy Diagnosis of Oral Carcinoma by the Combination of Morphological and Spectral Methods Based on Embedded Relay Lens Microscopic Hyperspectral Imaging System.

Cytopathological examination through biopsy is very important for carcinoma detection. The embedded relay lens microscopic hyperspectral imaging system (ERL-MHIS) provides a morphological image of a biopsy sample and the spectrum of each pixel in the image simultaneously. Based on the ERL-MHIS, this work develops morphological and spectral methods to diagnose oral carcinoma biopsy. In morphological discrimination, the fractal dimension method is applied to differentiate between normal and abnormal tissues. In spectral identification, normal and cancerous cells are distinguished using five methods. However, the spectra of normal and cancerous cells vary with patient. The diagnostic performances of the five methods are thus not ideal. Hence, the proposed cocktail approach is used to determine the effectiveness of the spectral methods in correlating with the sampling conditions. And then we use a combination of effective spectral methods according to the sample conditions for diagnosing a sample. A total of 68 biopsies from 34 patients are analyzed using the ERL-MHIS. The results demonstrate a sensitivity of 90 ± 4.53 % and a specificity of 87.8 ± 5.21 %. Furthermore, in our survey, this system is the first time utilized to study oral carcinoma biopsies.