Impact of lymph node dissection on surgical and oncological outcomes in patients undergoing robot-assisted radical cystectomy for bladder cancer: a multicenter retrospective study.

This study was performed to clarify the therapeutic and diagnostic roles of lymph node dissection (LND) by examining the impact of LND and lymph node yield (LNY) on oncological outcomes in patients undergoing robot-assisted radical cystectomy (RARC). Between 2014 and 2021, 216 patients underwent LND during RARC at Tokushima University Hospital and affiliated hospitals. Among the 216 patients, we compared 115 patients with an LNY of ≥ 20 and 101 with an LNY of < 20 to investigate the impact of LNY on surgical and oncological outcomes. Furthermore, we investigated the impact of LNY and the extent of LND on oncological outcomes by dividing the extent of LND into two groups (standard and extended). The 3-year rates of overall survival (OS) (p = 0.256), cancer-specific survival (CSS) (p = 0.791), and recurrence-free survival (RFS) (p = 0.953) did not differ between the two groups divided by the LNY. A higher LNY was associated with a significantly higher lymph node positivity rate (p = 0.020). The incidence of LND-related major complications was not significantly different between the two groups (p = 0.910). The 3-year survival rates did not differ between the two groups divided by the extent of LND: OS (p = 0.366), CSS (p = 0.814), and RFS (p = 0.689). The LNY and extent of LND were not associated with oncological outcomes in patients undergoing LND during RARC, whereas a higher LNY was associated with lymph node positivity. In the era of adjuvant therapy with immune checkpoint inhibitors, LND during RARC has an important diagnostic role in the detection of pathological node positivity.

Changes in Prevalence and Health Checkup Coverage Rate of Chronic Kidney Disease (CKD) after Introduction of Prefecture-Wide CKD Initiative: Results of the Kagawa Association of CKD Initiatives.

The National Health Insurance (NHI) special health checkup system in Japan targets the NHI population aged 40-74 years. Since 2015, the Kagawa NHI special health checkup was initiated in a prefecture-wide chronic kidney disease (CKD) initiative, including renal examination as an essential item in NHI health checkups. Here, we aimed to investigate the effects of the prefecture-wide CKD initiative. We conducted a retrospective cohort survey using the Kagawa National Health Insurance database created by the Kagawa National Health Insurance Organization. Results of the NHI health checkup (2015-2019) and prefecture-wide outcomes (2013-2019) were analyzed. The prevalence of CKD among examinees who underwent the NHI health checkup increased from 17.7% in 2015 to 23.2% in 2019. The percentage of examinees who completed a medical visit was 29.4% in 2015. After initiation of the initiative, the NHI health checkup coverage rate increased significantly, from a mean (standard deviation) of 40.8% (0.4%) to 43.2% (1.1%) (p = 0.04). After the start of the CKD initiative, we found an increase in the prevalence of CKD and the NHI health checkup coverage rate.

Signal-to-background ratio and lateral resolution in deep tissue imaging by optical coherence microscopy in the 1700 nm spectral band.

We quantitatively investigated the image quality in deep tissue imaging with optical coherence microscopy (OCM) in the 1700 nm spectral band, in terms of the signal-to-background ratio (SBR) and lateral resolution. In this work, to demonstrate the benefits of using the 1700 nm spectral band for OCM imaging of brain samples, we compared the imaging quality of OCM en-face images obtained at the same position by using a hybrid 1300 nm/1700 nm spectral domain (SD) OCM system with shared sample and reference arms. By observing a reflective resolution test target through a 1.5 mm-thick tissue phantom, which had a similar scattering coefficient to brain cortex tissue, we confirmed that 1700 nm OCM achieved an SBR about 6-times higher than 1300 nm OCM, although the lateral resolution of the both OCMs was similarly degraded with the increase of the imaging depth. Finally, we also demonstrated high-contrast deep tissue imaging of a mouse brain at a depth up to 1.8 mm by using high-resolution 1700 nm SD-OCM.

Visible-wavelength two-photon excitation microscopy with multifocus scanning for volumetric live-cell imaging.

Two-photon excitation microscopy is one of the key techniques used to observe three-dimensional (3-D) structures in biological samples. We utilized a visible-wavelength laser beam for two-photon excitation in a multifocus confocal scanning system to improve the spatial resolution and image contrast in 3-D live-cell imaging. Experimental and numerical analyses revealed that the axial resolution has improved for a wide range of pinhole sizes used for confocal detection. We observed the 3-D movements of the Golgi bodies in living HeLa cells with an imaging speed of 2 s per volume. We also confirmed that the time-lapse observation up to 8 min did not cause significant cell damage in two-photon excitation experiments using wavelengths in the visible light range. These results demonstrate that multifocus, two-photon excitation microscopy with the use of a visible wavelength can constitute a simple technique for 3-D visualization of living cells with high spatial resolution and image contrast.

Multi-Purpose Nanovoid Array Plasmonic Sensor Produced by Direct Laser Patterning.

We demonstrate a multi-purpose plasmonic sensor based on a nanovoid array fabricated via inexpensive and highly-reproducible direct femtosecond laser patterning of thin glass-supported Au films. The proposed nanovoid array exhibits near-IR surface plasmon (SP) resonances, which can be excited under normal incidence and optimised for specific applications by tailoring the array periodicity, as well as the nanovoid geometric shape. The fabricated SP sensor offers competitive sensitivity of ≈ 1600 nm/RIU at a figure of merit of 12 in bulk refractive index tests, as well as allows for identification of gases and ultra-thin analyte layers, making the sensor particularly useful for common bioassay experiments. Moreover, isolated nanovoids support strong electromagnetic field enhancement at lattice SP resonance wavelength, allowing for label-free molecular identification via surface-enhanced vibration spectroscopy.

Investigation of dispersion-managed, polarization-maintaining Er-doped figure-nine ultrashort-pulse fiber laser.

Figure-nine fiber lasers can realize all-polarization-maintaining, self-started, highly stable mode-locked laser sources, and are very attractive for applications such as optical frequency combs, metrology, etc. In this work, we investigated a dispersion-managed, polarization-maintaining, Er-doped, ultrashort-pulse figure-nine fiber laser both experimentally and numerically. Stable, self-started, passive mode-locking operation was achieved in a wide net cavity dispersion region, covering the soliton, stretched pulse, and dissipative soliton mode-locking regimes. A 132 fs ultrashort pulse with spectral width of 46 nm was obtained in the stretched pulse mode-locking regime. The initial mode-locking process and dynamics inside the cavity, in addition to the fundamental characteristics of the output pulses, were examined via numerical analysis. Owing to the asymmetric configuration, the propagation behaviors were different between the two counter-propagation directions. It was found that a large breathing had already started before the passive mode-locking point in stretched pulse mode-locking operation. Intense overshoots were also observed at the beginning of passive mode-locking. Numerical results were almost in agreement with the experimental ones.

High-spatial-resolution deep tissue imaging with spectral-domain optical coherence microscopy in the 1700-nm spectral band.

We present three-dimensional (3-D) high-resolution spectral-domain optical coherence microscopy (SD-OCM) by using a supercontinuum (SC) fiber laser source with 300-nm spectral bandwidth (full-width at half-maximum) in the 1700-nm spectral band. By using low-coherence interferometry with SC light and a confocal detection scheme, we realized lateral and axial resolutions of 3.4 and 3.8 µm in tissue (n = 1.38), respectively. This is, to the best of our knowledge, the highest 3-D spatial resolution reported among those of Fourier-domain optical coherence imaging techniques in the 1700-nm spectral band. In our SD-OCM, to enhance the imaging depth, a full-range method was implemented, which suppressed the formation of a coherent ghost image and allowed us to set the zero-delay position inside the samples. We demonstrated the 3-D high-resolution imaging capability of 1700-nm SD-OCM through the measurement of an interference signal from a mirror surface and imaging of a single 200-nm polystyrene bead and a pig thyroid gland. Deep tissue imaging at a depth of up to 1.8 mm was also demonstrated. This is the first demonstration of 3-D high-resolution SD-OCM in the 1700-nm spectral band.

Excitation of erbium-doped nanoparticles in 1550-nm wavelength region for deep tissue imaging with reduced degradation of spatial resolution.

Rare-earth-doped nanoparticles are one of the emerging probes for bioimaging due to their visible-to-near-infrared (NIR) upconversion emission via sequential single-photon absorption at NIR wavelengths. The NIR-excited upconversion property and high photostability make this probe appealing for deep tissue imaging. So far, upconversion nanoparticles include ytterbium ions (Yb3 + ) codoped with other rare earth ions, such as erbium (Er3 + ) and thulium (Tm3 + ). In these types of upconversion nanoparticles, through energy transfer from Yb3 + excited with continuous wave light at a wavelength of 980 nm, upconversion emission of the other rare earth dopants is induced. We have found that the use of the excitation of Er3 + in the 1550-nm wavelength region allows us to perform deep tissue imaging with reduced degradation of spatial resolution. In this excitation­emission process, three and four photons of 1550-nm light are sequentially absorbed, and Er3 + emits photons in the 550- and 660-nm wavelength regions. We demonstrate that, compared with the case using 980-nm wavelength excitation, the use of 1550-nm light enables us to moderate degradation of spatial resolution in deep tissue imaging due to the lower light scattering coefficient compared with 980-nm light. We also demonstrate that live cell imaging is feasible with this 1550 nm excitation.

The effects of a participatory structured group educational program on the development of CKD: a population-based study.

BACKGROUND: The type of lifestyle guidance that is effective for preventing development of chronic kidney disease (CKD) is unknown. Here, we aim to investigate the effects of a participatory structured group education (SGE) program on the development of CKD in a population-based study. METHODS: We retrospectively analyzed 1060 adult special health check-up examinees with CKD. Examinees with an estimated glomerular filtration rate (eGFR) from 50 to 60 mL/min/1.73 m2 and/or proteinuria 1+ were encouraged to attend an SGE program. The SGE program included participatory small group discussions on the attendees' remaining risk factors. The primary outcome of this study was the change in eGFR per year. RESULTS: The changes in eGFR in examinees who attended the SGE program (n = 209, + 2.9 mL/min/1.73 m2 [95% confidence interval (CI) + 1.9 to + 3.9]) significantly improved compared with control (n = 383, + 1.2 mL/min/1.73 m2 [95% CI + 0.5 to + 1.9], p = 0.006). Attending an SGE program was independently and positively related to the changes in eGFR at 1 year after attendance, after adjusting for classical covariates (ß = 1.55 [95% CI 0.37-2.73], p = 0.01). Attending an SGE program was effective for the examinees with a lower eGFR compared with those with only proteinuria. CONCLUSIONS: Our SGE program showed the beneficial effects of preventing the development of CKD, independent of classical factors. The type of SGE program that is more effective for preventing development of CKD should be investigated in a long-term analysis.

Axial resolution and signal-to-noise ratio in deep-tissue imaging with 1.7-µm high-resolution optical coherence tomography with an ultrabroadband laser source.

We investigated the axial resolution and signal-to-noise ratio (SNR) characteristics in deep-tissue imaging by 1.7-µm optical coherence tomography (OCT) with the axial resolution of 4.3 µm in tissue. Because 1.7-µm OCT requires a light source with a spectral width of more than 300 nm full-width at half maximum to achieve such high resolution, the axial resolution in the tissue might be degraded by spectral distortion and chromatic dispersion mismatching between the sample and reference arms. In addition, degradation of the axial resolution would also lead to reduced SNR. Here, we quantitatively evaluated the degradation of the axial resolution and the resulting decrease in SNR by measuring interference signals through a lipid mixture serving as a turbid tissue phantom with large scattering and absorption coefficients. Although the axial resolution was reduced by a factor of ∼6 after passing through a 2-mm-thick tissue phantom, our result clearly showed that compensation of the dispersion mismatching allowed us to achieve an axial resolution of 4.3 µm in tissue and improve the SNR by ∼5 dB compared with the case where dispersion mismatching was not compensated. This improvement was also confirmed in the observation of a hamster's cheek pouch in a buffer solution.

[MICROCYSTIC VARIANT OF UROTHELIAL CARCINOMA IN URINARY BLADDER: A CASE REPORT].

In the present report, we describe a patient with microcystic variant of urothelial carcinoma in urinary bladder. In March 2016, a 71-year-old man presented with bladder tumors found incidentally by ultrasonography. Cystoscopy and contrast-enhanced computed tomography (CT) revealed multiple invasive tumor of posterior wall, with a maximum diameter of 33 mm. Transurethral resection (TUR) of bladder tumors was performed. Pathological diagnosis was urothelial carcinoma, high grade, T2 or more. Invasive urothelial carcinoma was diagnosed and laparoscopic radical cystectomy with orthotopic neobladder was performed accordingly in April 2016. Pathological findings indicated a diagnosis of microcystic variant of urothelial carcinoma. At present, six months after surgery, the patient remains free of recurrence and metastasis. Here we review the characteristics of 4 microcystic variant of urothelial carcinoma cases reported in Japan.

[ROBOT-ASSISTED RADICAL PROSTATECTOMY FOR MEN AGE 75 AND OLDER].

(Objectives) Surgical treatment prostate cancer in elderly patients is controversial. However, robot-assisted radical prostatectomy (RARP) is a less invasive procedure than conventional surgery. Therefore, we perform RARP for elderly patients whose general condition is good (Performance status ≤1). The aim of this study is to evaluate surgical, oncological and functional outcomes for RARP in men age 75 and older. (Patients and methods) From July 2013 to April 2016, 300 patients underwent RARP at our institution. They were divided into two groups: an older patient group (≥75 years) and a younger patient group (<75 years). Treatment outcomes for each group, including surgical, oncological and functional outcomes, were compared. (Results) There were no statistically significant differences in surgical outcomes with the exception of nerve sparing rates (older patients: 5.9% vs. younger patients: 17.7%, P=0.0192). Importantly, intra- and postoperative complication rates were similar in both groups (minor complication: 7.4% vs. 3.9%, P=0.322, major complication: 0.0% vs. 2.2%, P=0.592). Regarding oncological outcomes, including positive surgical margin rate and PSA failure (PSA>0.2 ng/ml) at 12 months after surgery, no significant differences existed. Lastly, functional outcomes between the groups, including continence (≤1 pads/day) at 12 months after surgery, had no significant differences. (Conclusions) Our data suggests that RARP can be performed safely for men age 75 and older, and can become a good option for older patients with prostate cancer.

Optical coherence microscopy in 1700 nm spectral band for high-resolution label-free deep-tissue imaging.

Optical coherence microscopy (OCM) is a label-free, high-resolution, three-dimensional (3D) imaging technique based on optical coherence tomography (OCT) and confocal microscopy. Here, we report that the 1700-nm spectral band has the great potential to improve the imaging depth in high-resolution OCM imaging of animal tissues. Recent studies to improve the imaging depth in OCT revealed that the 1700-nm spectral band is a promising choice for imaging turbid scattering tissues due to the low attenuation of light in the wavelength region. In this study, we developed high-resolution OCM by using a high-power supercontinuum source in the 1700-nm spectral band, and compared the attenuation of signal-to-noise ratio between the 1700-nm and 1300-nm OCM imaging of a mouse brain under the condition of the same sensitivity. The comparison clearly showed that the 1700-nm OCM provides larger imaging depth than the 1300-nm OCM. In this 1700-nm OCM, the lateral resolution of 1.3 µm and the axial resolution of 2.8 µm, when a refractive index was assumed to be 1.38, was achieved.

Ultrasmall all-optical plasmonic switch and its application to superresolution imaging.

Because of their exceptional local-field enhancement and ultrasmall mode volume, plasmonic components can integrate photonics and electronics at nanoscale, and active control of plasmons is the key. However, all-optical modulation of plasmonic response with nanometer mode volume and unity modulation depth is still lacking. Here we show that scattering from a plasmonic nanoparticle, whose volume is smaller than 0.001 µm(3), can be optically switched off with less than 100 µW power. Over 80% modulation depth is observed, and shows no degradation after repetitive switching. The spectral bandwidth approaches 100 nm. The underlying mechanism is suggested to be photothermal effects, and the effective single-particle nonlinearity reaches nearly 10(-9) m(2)/W, which is to our knowledge the largest record of metallic materials to date. As a novel application, the non-bleaching and unlimitedly switchable scattering is used to enhance optical resolution to λ/5 (λ/9 after deconvolution), with 100-fold less intensity requirement compared to similar superresolution techniques. Our work not only opens up a new field of ultrasmall all-optical control based on scattering from a single nanoparticle, but also facilitates superresolution imaging for long-term observation.

Measurement of Scattering Nonlinearities from a Single Plasmonic Nanoparticle.

Plasmonics, which are based on the collective oscillation of electrons due to light excitation, involve strongly enhanced local electric fields and thus have potential applications in nonlinear optics, which requires extraordinary optical intensity. One of the most studied nonlinearities in plasmonics is nonlinear absorption, including saturation and reverse saturation behaviors. Although scattering and absorption in nanoparticles are closely correlated by the Mie theory, there has been no report of nonlinearities in plasmonic scattering until very recently. Last year, not only saturation, but also reverse saturation of scattering in an isolated plasmonic particle was demonstrated for the first time. The results showed that saturable scattering exhibits clear wavelength dependence, which seems to be directly linked to the localized surface plasmon resonance (LSPR). Combined with the intensity-dependent measurements, the results suggest the possibility of a common mechanism underlying the nonlinear behaviors of scattering and absorption. These nonlinearities of scattering from a single gold nanosphere (GNS) are widely applicable, including in super-resolution microscopy and optical switches. In this paper, it is described in detail how to measure nonlinearity of scattering in a single GNP and how to employ the super-resolution technique to enhance the optical imaging resolution based on saturable scattering. This discovery features the first super-resolution microscopy based on nonlinear scattering, which is a novel non-bleaching contrast method that can achieve a resolution as low as l/8 and will potentially be useful in biomedicine and material studies.

Visible-wavelength two-photon excitation microscopy for fluorescent protein imaging.

The simultaneous observation of multiple fluorescent proteins (FPs) by optical microscopy is revealing mechanisms by which proteins and organelles control a variety of cellular functions. Here we show the use of visible-light based two-photon excitation for simultaneously imaging multiple FPs. We demonstrated that multiple fluorescent targets can be concurrently excited by the absorption of two photons from the visible wavelength range and can be applied in multicolor fluorescence imaging. The technique also allows simultaneous single-photon excitation to offer simultaneous excitation of FPs across the entire range of visible wavelengths from a single excitation source. The calculation of point spread functions shows that the visible-wavelength two-photon excitation provides the fundamental improvement of spatial resolution compared to conventional confocal microscopy.

A fast- and positively photoswitchable fluorescent protein for ultralow-laser-power RESOLFT nanoscopy.

Fluorescence nanoscopy has revolutionized our ability to visualize biological structures not resolvable by conventional microscopy. However, photodamage induced by intense light exposure has limited its use in live specimens. Here we describe Kohinoor, a fast-switching, positively photoswitchable fluorescent protein, and show that it has high photostability over many switching repeats. With Kohinoor, we achieved super-resolution imaging of live HeLa cells using biocompatible, ultralow laser intensity (0.004 J/cm(2)) in reversible saturable optical fluorescence transition (RESOLFT) nanoscopy.

Point spread function analysis with saturable and reverse saturable scattering.

Nonlinear plasmonics has attracted a lot of interests due to its wide applications. Recently, we demonstrated saturation and reverse saturation of scattering from a single plasmonic nanoparticle, which exhibits extremely narrow side lobes and central peaks in scattering images [ACS Photonics 1(1), 32 (2014)]. It is desirable to extract the reversed saturated part to further enhance optical resolution. However, such separation is not possible with conventional confocal microscope. Here we combine reverse saturable scattering and saturated excitation (SAX) microscopy. With quantitative analyses of amplitude and phase of SAX signals, unexpectedly high-order nonlinearities are revealed. Our result provides greatly reduced width in point spread function of scattering-based optical microscopy. It will find applications in not only nonlinear material analysis, but also high-resolution biomedical microscopy.

Introduction to super-resolution microscopy.

In this review, we introduce the principles of spatial resolution improvement in super-resolution microscopies that were recently developed. These super-resolution techniques utilize the interaction of light and fluorescent probes in order to break the diffraction barrier that limits spatial resolution. The imaging property of each super-resolution technique is also compared with the corresponding conventional one. Typical applications of the super-resolution techniques in biological research are also introduced.

Saturated excitation microscopy with optimized excitation modulation.

Saturated excitation (SAX) microscopy utilizes the nonlinear relation between fluorescence emission and excitation under saturated excitation to improve the spatial resolution of confocal microscopy. In this study, we theoretically and experimentally investigate the saturation of fluorescence excitation under modulated excitation to optimize the excitation conditions for SAX microscopy. Calculation of the relationships between fluorescence and excitation intensity with different modulation frequencies reveals that the lifetime of the triplet state of the fluorescent probe strongly affects the strength of the demodulated fluorescence signals. We also find that photobleaching shows little dependence on the modulation frequency. These investigations allow us to determine the optimum excitation conditions, that is, the conditions providing sufficient fluorescence saturation without strong photobleaching. For a sample stained with ATTO Rho6G phalloidin, we estimate the optimal excitation conditions, which are produced with 50 kHz excitation modulation and a 50 µsec pixel dwell time, and successfully perform three-dimensional imaging with sub-diffraction resolution.