Infantile Myofibromatosis of the Femoral Neck: A Case Report.

CASE: A 15-month-old boy who was being followed for developmental dysplasia of the hip because of breech presentation was discovered to have a solitary infantile myofibroma in the left femoral neck. The patient was avoiding weight-bearing on the affected extremity; thus, stabilization of the femoral neck was performed using a proximal femur locking plate. Postoperatively, he achieved all gross motor developmental milestones. CONCLUSION: This report is the first to describe a solitary infantile myofibroma in the femoral neck and demonstrates the utility of operative stabilization of these lesions.

Full-Quantum Treatment of Molecular Systems Confirms Novel Supracence Photonic Properties.

Our understanding of molecules has stagnated at a single quantum system, with atoms as Newtonian particles and electrons as quantum particles. Here, however, we reveal that both atoms and electrons in a molecule are quantum particles, and their quantum-quantum interactions create a previously unknown, newfangled molecular property-supracence. Molecular supracence is a phenomenon in which the molecule transfers its potential energy from quantum atoms to photo-excited electrons so that the emitted photon has more energy than that of the absorbed one. Importantly, experiments reveal such quantum energy exchanges are independent of temperature. When quantum fluctuation results in absorbing low-energy photons, yet still emitting high-energy photons, supracence occurs. This report, therefore, reveals novel principles governing molecular supracence via experiments that were rationalized by full quantum (FQ) theory. This advancement in understanding predicts the super-spectral resolution of supracence, and molecular imaging confirms such innovative forecasts using closely emitting rhodamine 123 and rhodamine B in living cell imaging of mitochondria and endosomes.

Cooperatively assembled liquid crystals enable temperature-controlled Förster resonance energy transfer.

Balancing the rigidity of a π-conjugated structure for strong emission and the flexibility of liquid crystals for self-assembly is the key to realizing highly emissive liquid crystals (HELCs). Here we show that (1) integrating organization-induced emission into dual molecular cooperatively-assembled liquid crystals, (2) amplifying mesogens, and (3) elongating the spacer linking the emitter and the mesogen create advanced materials with desired thermal-optical properties. Impressively, assembling the fluorescent acceptor Nile red into its host donor designed according to the aforementioned strategies results in a temperature-controlled Förster resonance energy transfer (FRET) system. Indeed, FRET exhibits strong S-curve dependence as temperature sweeps through the liquid crystal phase transformation. Such thermochromic materials, suitable for dynamic thermo-optical sensing and modulation, are anticipated to unlock new and smart approaches for controlling and directing light in stimuli-responsive devices.

Creating an Assessment Tool for Clinical Musculoskeletal Knowledge.

ABSTRACT: Studies show that medical school curriculums do not prepare graduates to manage the most common musculoskeletal (MSK) injuries they will encounter in the outpatient setting. The authors proposed a new multiple-choice assessment to identify individual and program deficiencies for curriculum improvement. A multiple-choice MSK assessment tool was administered to learners at various stages of training. Students took the examination after completing their core clerkship year; residents took the examination near the beginning of their respective academic year. Outcome measures included average examination score, percent correct for each question, and overall examination reliability. Average examinees scores were 75.6% with higher scores based on training years. No statistical differences were found between MD/DO, male/female, or military/civilian examinees. The tool was found to be a statistically valid method of determining cognitive knowledge in basic MSK topics, identifying individual deficiencies, and highlighting gaps in training programs' MSK curriculums.

Molecular Supracence Resolving Eight Colors in 300-nm Width: Unprecedented Spectral Resolution.

Monitoring multiple molecular probes simultaneously establishes their correlations and reveals the holistic mechanism. Current fluorescence imaging, however, is limited to about four colors because of typically circa 100-nm spectral width. Herein, we show that molecular supracence imparts superior spectral resolution, resolving eight colors in 300-nm width, about 37.5-nm per color. A recently discovered light-molecule interacting phenomenon, supracence only measures molecular emission above its excitation energy due to entanglement between atomic quantum system and electronic quantum system. As such, supracence takes advantage of sharp spectral edge of a single pathway and excitation specificity to produce narrow bands, whereas fluorescence has to deal with multiple pathways spilling out low-energy long tail, that causes poor resolution. Thus, supracence enables myriad innovative molecular spectroscopy and microscopic imaging with profound impact broadly.

Discovery of a New Light-Molecule Interaction: Supracence Reveals What Is Missing in Fluorescence Imaging.

The currently understood principles about light-molecule interactions are limited, and thus scientific scope beyond current theories is rarely harvested. Herein we demonstrate supracence phenomena, in which the emitted photons have more energy than the absorbed photons. The extra energy comes from couplings of the absorbed and emitted photon to molecular phonons, whose potentials are constantly exchanging with molecular quantum energy and the environment. Thus, supracence is a linear optical process rather than a nonlinear optical process, such as second harmonic generation. Because supracence results in cooled molecular phonons and thus cooled molecules, behavior opposite to that of hot fluorescing emitters is expected. This report reveals certain supracence principles while contrasting fluorescence with supracence in high-resolution imaging.

Chemomechanical-force-induced folding-unfolding directly controls distinct fluorescence dual-color switching.

Folding-unfolding imparts fluorescence dual color switching, thus a novel concept to switch fluorescence between two distinct colors while avoiding traditional bond rupturing and bond forming in photoswitching. Because folding and unfolding minimize the wear and tear on molecular structures, the new systems have excellent reversibility and fatigue resistance.

Photoswitching Near-Infrared Fluorescence from Polymer Nanoparticles Catapults Signals over the Region of Noises and Interferences for Enhanced Sensitivity.

As a very sensitive technique, photoswitchable fluorescence not only gains ultrasensitivity but also imparts many novel and unexpected applications. Applications of near-infrared (NIR) fluorescence have demonstrated low background noises, high tissue-penetrating ability, and an ability to reduce photodamage to live cells. Because of these desired features, NIR-fluorescent dyes have been the premium among fluorescent dyes, and probes with photoswitchable NIR fluorescence are even more desirable for enhanced signal quality in the emerging optical imaging modalities but rarely used because they are extremely challenging to design and construct. Using a spiropyran derivative functioning as both a photoswitch and a fluorophore to launch its periodically modulated red fluorescence excitation energy into a NIR acceptor, we fabricated core-shell polymer nanoparticles exhibiting a photoswitchable fluorescence signal within the biological window (∼700-1000 nm) with a peak maximum of 776 nm. Live cells constantly synthesize new molecules, including fluorescent molecules, and also endocytose exogenous particles, including fluorescent particles. Upon excitation at different wavelengths, these fluorescent species bring about background noises and interferences covering nearly the whole visible region and therefore render many intracellular targets unaddressable. The oscillating NIR fluorescence signal with an on/off ratio of up to 67 that the polymer nanoparticles display is beyond the typical background noises and interferences, thus producing superior sharpness, reliability, and signal-to-noise ratios in cellular imaging. Taking these salient features, we anticipate that these types of nanoparticles will be useful for in vivo imaging of biological tissue and other complex specimens, where two-photon activation and excitation are used in combination with NIR-fluorescence photoswitching.

Photoswitchable fluorescent nanoparticles and their emerging applications.

Although fluorescence offers ultrasensitivity, real-world applications of fluorescence techniques encounter many practical problems. As a noninvasive means to investigate biomolecular mechanisms, pathways, and regulations in living cells, the intrinsic heterogeneity and inherent complexity of biological samples always generates optical interferences such as autofluorescence. Therefore, innovative fluorescence technologies are needed to enhance measurement reliability while not compromising sensitivity. In this review, we present current strategies that use photoswitchable nanoparticles to address these real-world challenges. The unique feature in these photoswitchable nanoparticles is that fundamental molecular photoswitches are playing the critical role of fluorescence modulation rather than traditional methods like modulating the light source. As a result, new innovative technologies that have recently emerged include super-resolution imaging, frequency-domain imaging, antiphase dual-color correlation, etc. Some of these methods improve imaging resolution down to the nanometer level, while others boost the detection sensitivity by orders of magnitude and confirm the nanoparticle probes unambiguously. These enhancements, which are not possible with non-photoswitching molecular probes, are the central topics of this review.

Antiphase dual-color correlation in a reactant-product pair imparts ultrasensitivity in reaction-linked double-photoswitching fluorescence imaging.

A pair of reversible photochemical reactions correlates their reactant and product specifically, and such a correlation uniquely distinguishes their correlated signal from others that are not linked by this reversible reaction. Here a nanoparticle-shielded fluorophore is photodriven to undergo structural dynamics, alternating between a green-fluorescence state and a red-fluorescence state. As time elapses, the fluorophore can be in either state but not both at the same time. Thus, the red fluorescence is maximized while the green fluorescence is minimized and vice versa. Such an antiphase dual-color (AD) corelationship between the red and green fluorescence maxima as well as between their minima can be exploited to greatly improve the signal-to-noise ratio, thus enhancing the ultimate detection limit. Potential benefits of this correlation include elimination of all interferences originating from single-color dyes and signal amplification of AD photoswitching molecules by orders of magnitude.

Reversible photoswitching specifically responds to mercury(II) ions: the gated photochromism of bis(dithiazole)ethene.

Photoswitching of bis(dithiazole)ethene can be regulated by Hg(II) ions and EDTA in a "lock-and-unlock" manner. The molecular photoswitch provides an enzyme-like binding pocket that selectively binds specifically to mercury ions, thus modulating the degree of photoswitching in its presence.

Chromophoric and dendritic phosphoramidites enable construction of functional dendrimers with exceptional brightness and water solubility.

The fluorescence brightness of a molecular probe determines whether it can be effectively measured and its water solubility dictates if it can be applied in real-world biological systems. However, molecules brighter than the most efficient fluorescent dyes or particles brighter than quantum dots are hard to come by, especially when they must also be soluble in water. In this report, chromophoric phosphoramidites are used in a solid-state synthesis to construct functional dendrimers. When highly twisted chromophores are chosen and the proper spacers and dendrons are introduced, the resultant dendrimers emit exceptionally bright fluorescence. Chromophores, spacers, and dendrons are stitched together by efficient phosphoramidite reagents, which afford high-yield water-soluble phosphodiester linkages after deprotection. The resulting water-soluble dendrimers are exceptionally bright.

Quantitative photoswitching in bis(dithiazole)ethene enables modulation of light for encoding optical signals.

Using one ray of light to encode another ray of light is highly desirable because information in optical format can be directly transferred from one beam to another without converting back to the electronic format. One key medium to accomplish such an amazing task is photoswitchable molecules. Using bis(dithiazole)ethene that can be photoswitched between its ring-open and ring-closed states quantitatively with excellent fatigue resistance and high thermal stability, it is shown that quantitative photoreversibility allowed the photoswitching light to control other light travelling through the photoswitchable medium, a phenomenon of transferring information encoded in one light ray to others, thus imparting photo-optical modulation on the orthogonal light beam.

Understanding super-resolution nanoscopy and its biological applications in cell imaging.

Optical microscopy has been an ideal tool for studying phenomena in live cells because visible light at reasonable intensity does not perturb much of the normal biological functions. However, optical resolution using visible light is significantly limited by the wavelength. Overcoming this diffraction-limit barrier will reveal biological mechanisms, cellular structures, and physiological processes at a nanometer scale, orders of magnitude lower than current optical microscopy. Although this appears to be a daunting task, recently developed photoswitchable probes enable reconstruction of individual images into a super-resolution image, thus the emergence of nanoscopy. Harnessing the resolution power of nanoscopy, we report here nano-resolution fluorescence imaging of microtubules and their network structures in biological cells. The super-resolution nanoscopy successfully resolved nanostructures of a microtubule network-a daunting task that cannot be completed using conventional wide-field microscopy.

Photoswitching-enabled novel optical imaging: innovative solutions for real-world challenges in fluorescence detections.

Because of its ultrasensitivity, fluorescence offers a noninvasive means to investigate biomolecular mechanisms, pathways, and regulations in living cells, tissues, and animals. However, real-world applications of fluorescence technologies encounter many practical challenges. For example, the intrinsic heterogeneity of biological samples always generates optical interferences. High background such as autofluorescence can often obscure the desired signals. Finally, the wave properties of light limit the spatial resolution of optical microscopy. The key to solving these problems involves using chemical structures that can modulate the fluorescence output. Photoswitchable fluorescent molecules that alternate their emissions between two colors or between bright-and-dark states in response to external light stimulation form the core of these technologies. For example, molecular fluorescence modulation can switch fluorophores on and off. This feature supports super-resolution, which enhances resolution by an order of magnitude greater than the longstanding diffraction-limit barrier. The reversible modulation of such probes at a particular frequency significantly amplifies the frequency-bearing target signal while suppressing interferences and autofluorescence. In this Account, we outline the fundamental connection between constant excitation and oscillating fluorescence. To create molecules that will convert a constant excitation into oscillating emission, we have synthesized photoswitchable probes and demonstrated them as proofs of concept in super-resolution imaging and frequency-domain imaging. First, we introduce the design of molecules that can convert constant excitation into oscillating emission, the key step in fluorescence modulation. Then we discuss various technologies that use fluorescence modulation: super-resolution imaging, dual-color imaging, phase-sensitive lock-in detection, and frequency-domain imaging. Finally, we present two biological applications to demonstrate the power of photoswitching-enabled fluorescence imaging. Because synthetic photoswitchable probes can be much smaller, more versatile, and more efficient at high-performance modulation experiments, they provide a complement to photoswitchable fluorescent proteins. Although new challenges remain, we foresee a bright future for photoswitching-enabled imaging and detection.

A self-assembly phase diagram from amphiphilic perylene diimides.

Supramolecular forces govern self-assembly and further determine the final morphologies of self-assemblies. However, how they control the morphology remains hitherto largely unknown. In this paper, we have discovered that the self-assembled nanostructures of rigid organic semiconductor chromophores can be finely controlled by the secondary forces by fine-tuning the surrounding environments. In particular, we used water/methanol/hydrochloric acid to tune the environment and observed five different phases that resulted from versatile molecular self-assemblies. The representative self-assembled nanostructures were nanotapes, nanoparticles and their 1D assemblies, rigid microplates, soft nanoplates, and hollow nanospheres and their 1D assemblies, respectively. The specific nanostructure formation is governed by the water fraction, R(w), and the concentration of hydrochloric acid, [HCl]. For instance, nanotapes formed at low [HCl] and R(w) values, whereas hollow nanospheres formed when either the HCl concentration is high, or the water fraction is low, or both. The significance of this paper is that it provides a useful phase diagram by using R(w) and [HCl] as two variables. Such a self-assembly phase diagram maps out the fine control that the secondary forces have on the self-assembled morphology, and thus allows one to guide the formation toward a desired nanostructure self-assembled from rigid organic semiconductor chromophores by simply adjusting the two key parameters of R(w) and [HCl].

Photoswitching-induced frequency-locked donor-acceptor fluorescence double modulations identify the target analyte in complex environments.

Precisely identifying biological targets and accurately extracting their relatively weak signals from complicated physiological environments represent daunting challenges in biological detection and biomedical diagnosis. Fluorescence techniques have become the method of choice and offer minimally invasive and ultrasensitive detections, thus, providing a wealth of information regarding the biological mechanisms in living systems. Despite fluorescence analysis has advanced remarkably, conventional detections still encounter considerable limitations. This stems from the fact that the fluorescence intensity signal (I) is sensitive and liable to numerous external factors including temperature, light source, medium characteristics, and dye concentration. The interferences exasperatingly undermine the precision of measurements, and frequently render the signal undetectable. For example, fluorescence from single-molecule emitters can be measured on glass substrates under optimum conditions, but single-molecule events in complicated physiological environments such as live cells can hardly be detected because of autofluorescence interference and other factors. Furthermore, traditional intensity (I) and wavelength (λ) measurements do not reveal the interactive nature between the donor and the acceptor. Thus, innovative detection strategies to circumvent these aforementioned limitations of the conventional techniques are critically needed. With the use of photoswitching-induced donor-acceptor-fluorescence double modulations, we present a novel strategy that introduces three additional physical parameters: modulation amplitude (A), phase shift (ΔΦ), and lock-in frequency (ω), and demonstrate that such a strategy can circumvent the limitation of the conventional fluorescence detection techniques. Together, these five physical quantities (I, λ, A, ΔΦ, ω) reveal insightful information regarding molecular interactive strength between the probe and the analyte and enable extracting weak-fluorescence spectra from large interfering noises in complex environments.

Sequence-controlled oligomers fold into nanosolenoids and impart unusual optical properties.

Controlled syntheses give unique block oligomers with alternating flexible ethylene glycol and rigid perylenetetracarboxylic diimide (PDI) units. The number of rigid units vary from n=1 to 10. PDI units were stitched together by using efficient phosphoramidite chemistry. The resulting oligomers undergo folding in most solvents, including chloroform. In their ground state, these folded oligomers were characterized by using Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS), NMR spectroscopy, and electronic absorption spectroscopy. FTICR-MS revealed the exact masses of these sequence-controlled oligomers, which confirmed the chemical composition and validated the synthetic strategy. The NMR neighboring ring-current effect (NRE) indicates the formation of cofacial π stacks; the stacked aromatic rings have nearly coaxial alignment akin to a nanosoleniod. Nanosolenoidal shielding in π stacks causes all aromatic protons to shift upfield, whereas NOE in a cyclic hetero-chromophoric dimer supports a rotated, cofacial π-stacking orientation separated by about 3.5 Å. Electron-phonon coupling is much stronger than excitonic coupling in these self-folded PDI oligomers; thus, Franck-Condon factors dictate the observed spectral features in visible spectra. The absorbance spectrum exhibits weak hypochromism due to π stacking with increasing stacking units n. Finally, ab initio calculations support the experimental observations, indicating 3.5 Å cofacial spacing in which one molecule is rotated 30° from the eclipsed orientation and higher oligomers can adopt, without a compensating energy penalty, either the right/left-handed helices or the 1,3-eclipsed structures. Both theory and experiments validate the nano-π-solenoids and their novel photophysical properties.

Self-assembled hollow nanospheres strongly enhance photoluminescence.

We report that two molecular building blocks differ only by two protons, yet they form totally different nanostructures. The protonated one self-organized into hollow nanospheres (~200 nm), whereas the one without the protons self-assembled into rectangular plates. Consequently, the geometrically defined nanoassemblies exhibit radically different properties. As self-assembly directing units, protons impart ion-pairing and hydrogen-bonding probabilities. The plate-forming nanosystem fluoresces weakly, probably due to energy transfer among chromophores (Φ < 0.2), but the nanospheres emit strong yellow fluorescence (Φ ≈ 0.58-0.85).

Photoswitchable nanoprobes offer unlimited brightness in frequency-domain imaging.

A single probe has limited brightness in time-domain imaging and such limitation frequently renders individual molecules undetectable in the presence of interference or complex cellular structures. However, a single photoswitchable probe produces a signal, which can be separated from interference or noise using photoswitching-enabled Fourier transformation (PFT). As a result, the light-modulated probes can be made super bright in the frequency domain simply by acquiring more cycles in the time domain.