Actomyosin contractility drives bile regurgitation as an early response during obstructive cholestasis.

BACKGROUND & AIMS: A wide range of liver diseases manifest as biliary obstruction, or cholestasis. However, the sequence of molecular events triggered as part of the early hepatocellular homeostatic response in obstructive cholestasis is poorly elucidated. Pericanalicular actin is known to accumulate during obstructive cholestasis. Therefore, we hypothesized that the pericanalicular actin cortex undergoes significant remodeling as a regulatory response to obstructive cholestasis. METHODS: In vivo investigations were performed in a bile duct-ligated mouse model. Actomyosin contractility was assessed using sandwich-cultured rat hepatocytes transfected with various fluorescently labeled proteins and pharmacological inhibitors of actomyosin contractility. RESULTS: Actomyosin contractility induces transient deformations along the canalicular membrane, a process we have termed inward blebbing. We show that these membrane intrusions are initiated by local ruptures in the pericanalicular actin cortex; and they typically retract following repair by actin polymerization and actomyosin contraction. However, above a certain osmotic pressure threshold, these inward blebs pinch away from the canalicular membrane into the hepatocyte cytoplasm as large vesicles (2-8µm). Importantly, we show that these vesicles aid in the regurgitation of bile from the bile canaliculi. CONCLUSION: Actomyosin contractility induces the formation of bile-regurgitative vesicles, thus serving as an early homeostatic mechanism against increased biliary pressure during cholestasis. LAY SUMMARY: Bile canaliculi expand and contract in response to the amount of secreted bile, and resistance from the surrounding actin bundles. Further expansion due to bile duct blockade leads to the formation of inward blebs, which carry away excess bile to prevent bile build up in the canaliculi.

Linewidth suppression mechanism of self-injection locked single-frequency fiber laser.

Linewidth suppression mechanism of the self-injection locked single-frequency fiber laser (SFFL) is investigated theoretically and experimentally. An analytical model based on the semi-phenomenological approach is built up to characterize the optical feedback in SFFL. According to the theoretical prediction, the linewidth tends to be reduced with longer external cavity photon lifetime. Experimentally, a 200-Hz linewidth self-injection locked SFFL is achieved with 101 m long delay fiber, which agrees well with the theoretical simulation. The model provides a new perspective to understand the mechanism of linewidth reduction of self-injection locked SFFL.

High OSNR watt-level single-frequency one-stage PM-MOPA fiber laser at 1083 nm.

A 1.03 W optical signal-to-noise ratio (OSNR) of > 70 dB single-frequency polarization-maintained master-oscillator power amplifier (PM-MOPA) laser at 1083 nm was demonstrated. The seed laser of this laser system was a distributed Bragg reflector short cavity Yb-doped phosphate fiber oscillator. A one-stage core-pumped amplification configuration was employed, in which the typical gain is 9.7 dB and the optical-to-optical conversion efficiency is 68.7%. The estimated laser linewidth is less than 3.5 kHz, the measured polarization-extinction ratio is greater than 25 dB, and the obtained relative intensity noise of fiber laser for frequencies of over 2 MHz is less than -130 dB/Hz.

600-Hz linewidth short-linear-cavity fiber laser.

We proposed a short-linear-cavity (SLC) fiber laser based on a virtual-folded-ring (VFR) resonator and a fiber Bragg grating Fabry-Perot filter. Spatial hole burning effect was reduced by retarding the polarization state of the counter-propagating light waves utilizing the VFR structure. The photon lifetime of the resonator was extended due to the multi-reflection inside the FBG FP, which increased the intra-cavity power and relatively suppressed the contribution of phase diffusion from spontaneous emission. The relaxation oscillation frequency is around 160 kHz due to the slow light effect. The linewidth of the SLC fiber laser was measured to be less than 600 Hz.

1.95 µm kHz-linewidth single-frequency fiber laser using self-developed heavily Tm3+-doped germanate glass fiber.

We demonstrated a kHz-linewidth single-frequency laser at 1.95 µm using the self-developed heavily Tm(3+)-doped single-mode germanate glass fiber with the net gain coefficient of 2.3 dB per centimeter. The maximum output power of the stable single longitudinal mode continuous wave laser is over 200 mW. The slope efficiency measured versus the absorbed pump power is 34.8%, the signal-to-noise ratio is higher than 68 dB and laser linewidth is less than 7 kHz. A wavelength-tuning from 1949.55 to 1951.23 nm was also demonstrated based on changing the tension on the fiber Bragg grating outside the cavity.

A 1014 nm linearly polarized low noise narrow-linewidth single-frequency fiber laser.

We present the demonstration of a compact linearly polarized low noise narrow-linewidth single-frequency fiber laser at 1014 nm. The compact fiber laser is based on a 5-mm-long homemade Yb(3+)-doped phosphate fiber. Over 164 mW stable continuous-wave single transverse and longitudinal mode lasing at 1014 nm has been achieved. The measured relative intensity noise is less than -135 dB/Hz at frequencies of over 2.5 MHz. The signal-to-noise ratio of the laser is larger than 70 dB, and the linewidth is less than 7 kHz, while the obtained linear polarization extinction ratio is higher than 30 dB.

10.9 W kHz-linewidth one-stage all-fiber linearly-polarized MOPA laser at 1560 nm.

An all-fiber 10.9 W single-frequency one-stage linearly-polarized master-oscillator power amplifier (MOPA) laser at 1560 nm has been demonstrated. The laser linewidth is less than 3.5 kHz and the polarization-extinction ratio (PER) is greater than 24 dB. The measured signal-to-noise ratio (SNR) is higher than 70 dB and the optical-to-optical conversion efficiency is 29.5%. No obvious stimulated Brillouin scattering and the devastating effects of unwanted coupling light were observed.

Low noise single-frequency single-polarization ytterbium-doped phosphate fiber laser at 1083 nm.

We present a low noise single-frequency and single-polarization distributed Bragg reflector fiber laser at 1083 nm by using a 1.8 cm long newly developed ytterbium-doped phosphate single mode glass fiber. The maximum output power is more than 100 mW with a slope efficiency of >29.6%. The signal to noise ratio is higher than 61 dB and the laser linewidth of less than 2 kHz is estimated. The obtained relative intensity noise for frequencies of over 4.0 MHz is less than -150 dB/Hz, which approaches the shot noise limit. The achieved linear polarization extinction ratio is more than 30 dB.

Generation of mature kupffer cells from human induced pluripotent stem cells.

Liver macrophages, Kupffer cells (KCs), play a critical role in drug-induced liver injury (DILI) and liver diseases including cholestasis, liver fibrosis and viral hepatitis. Application of KCs in in vitro models of DILI and liver diseases is hindered due to limited source of human KCs. In vivo, KCs originate from MYB-independent macrophage progenitors, which differentiate into liver-specific macrophages in response to hepatic cues in the liver. Here, we recapitulated KCs ontogeny by differentiation of MYB-independent iPSCs to macrophage-precursors and exposing them to hepatic cues to generate iPSC-derived KCs (iKCs). iKCs expressed macrophage markers (CD11/CD14/CD68/CD163/CD32) at 0.3-5 folds of primary adult human KCs (pKCs) and KC-specific CLEC-4F, ID1 and ID3. iKCs phagocytosed and secreted IL-6 and TNFα upon stimulation at levels similar to pKCs but different from non-liver macrophages. Hepatocyte-iKCs co-culture model was more sensitive in detecting hepatotoxicity induced by inflammation-associated drugs, Acetaminophen and Trovafloxacin, and Chlorpromazine-induced cholestasis when compared to hepatocytes alone. Overall, iKCs were mature, liver-specific and functional. Furthermore, donor-matched iKCs and iPSC-hepatocyte co-culture exhibited minimal non-specific background response compared to donor-mismatched counterpart. iKCs offer a mature renewable human cell source for liver-specific macrophages, useful in developing in vitro model to study DILI and liver diseases such as cholestasis.

Deep learning enables automated scoring of liver fibrosis stages.

Current liver fibrosis scoring by computer-assisted image analytics is not fully automated as it requires manual preprocessing (segmentation and feature extraction) typically based on domain knowledge in liver pathology. Deep learning-based algorithms can potentially classify these images without the need for preprocessing through learning from a large dataset of images. We investigated the performance of classification models built using a deep learning-based algorithm pre-trained using multiple sources of images to score liver fibrosis and compared them against conventional non-deep learning-based algorithms - artificial neural networks (ANN), multinomial logistic regression (MLR), support vector machines (SVM) and random forests (RF). Automated feature classification and fibrosis scoring were achieved by using a transfer learning-based deep learning network, AlexNet-Convolutional Neural Networks (CNN), with balanced area under receiver operating characteristic (AUROC) values of up to 0.85-0.95 versus ANN (AUROC of up to 0.87-1.00), MLR (AUROC of up to 0.73-1.00), SVM (AUROC of up to 0.69-0.99) and RF (AUROC of up to 0.94-0.99). Results indicate that a deep learning-based algorithm with transfer learning enables the construction of a fully automated and accurate prediction model for scoring liver fibrosis stages that is comparable to other conventional non-deep learning-based algorithms that are not fully automated.