Differential Development of the Chordae Tendineae and Anterior Leaflet of the Bovine Mitral Valve.

There is increasing evidence that some adult mitral valve pathologies may have developmental origins involving errors in cell signaling and protein deposition during valvulogenesis. While early and late gestational stages are well-documented in zebrafish, chicks, and small mammalian models, longitudinal studies in large mammals with a similar gestational period to humans are lacking. Further, the mechanism of chordae tendineae formation and multiplication remains unclear. The current study presents a comprehensive examination of mitral anterior leaflet and chordae tendineae development in a bovine model (a large mammal with the same gestational period as humans). Remarkably distinct from small mammals, bovine development displayed early branched chordae, with increasing attachments only until birth, while the anterior leaflet grew both during gestation and postnatally. Chordae also exhibited accelerated collagen deposition, maturation, and crimp development during gestation. These findings suggest that the bovine anterior leaflet and chordae tendineae possess unique processes of development despite being a continuous collagenous structure and could provide greater insight into human valve development.

Ossicular Reconstruction With Various Degrees of Malleus Preservation in Patients With Chronic Otitis Media.

Objective: We examined the relationship between factors of middle ear conditions and the outcome of ossiculoplasty in chronic otitis media (COM) by measuring the improvement in the air-bone gap (ABG) and air conduction threshold (TAC). Methods: This retrospective study analyzed 76 patients (77 ears) who underwent ossiculoplasty from among 520 COM patients who underwent tympanoplasty based on the maximum preservation of the original ossicles. The reconstructed ossicular chain was performed by preserving or utilizing the remaining malleus in all cases with the presence of the malleus manubrium. Patients with eardrum adhesion, cholesteatoma, and cholesterol granuloma were defined as having a compromised middle ear condition (Group A), and those without as having an uncompromised middle ear condition (Group B). In each group, pure-tone audiometry was performed preoperatively and postoperatively, and improvements in the ABG and TAC were compared. The effects of the types of tympanoplasty and the method of ossiculoplasty (columella versus incus interposition) on postoperative ABG and TAC were also compared. Results: The postoperative ABG improvement in Group B was significantly higher than that in Group A [ß = 7.31, 95% confidence interval (CI) = 1.93-12.69, P < .05]. Type III minor columella tympanoplasty yielded significantly better results than type III major and type Vb tympanoplasty (ß = 11.42, 95% CI = 5.16-17.68, P < .01). There were no significant differences in the postoperative ABG or TAC between the reconstruction groups with and without preservation of malleus. Conclusions: Our results indicate that complex cases compromised by adhesions, cholesteatoma, and cholesterol granuloma have worse outcomes regarding hearing improvement and success rates, while those with intact stapes suprastructure have better outcomes. Malleus was maximally preserved in the patients of this study; however, this showed no significant prognostic benefit in hearing.

Isolated loss of the AUTS2 long isoform, brain-wide or targeted to Calbindin-lineage cells, generates a specific suite of brain, behavioral, and molecular pathologies.

Rearrangements within the AUTS2 region are associated with a rare syndromic disorder with intellectual disability, developmental delay, and behavioral abnormalities as core features. In addition, smaller regional variants are linked to wide range of neuropsychiatric disorders, underscoring the gene's essential role in brain development. Like many essential neurodevelopmental genes, AUTS2 is large and complex, generating distinct long (AUTS2-l) and short (AUTS2-s) protein isoforms from alternative promoters. Although evidence suggests unique isoform functions, the contributions of each isoform to specific AUTS2-linked phenotypes have not been clearly resolved. Furthermore, Auts2 is widely expressed across the developing brain, but cell populations most central to disease presentation have not been determined. In this study, we focused on the specific roles of AUTS2-l in brain development, behavior, and postnatal brain gene expression, showing that brain-wide AUTS2-l ablation leads to specific subsets of the recessive pathologies associated with mutations in 3' exons (exons 8-19) that disrupt both major isoforms. We identify downstream genes that could explain expressed phenotypes including hundreds of putative direct AUTS2-l target genes. Furthermore, in contrast to 3' Auts2 mutations which lead to dominant hypoactivity, AUTS2-l loss-of-function is associated with dominant hyperactivity and repetitive behaviors, phenotypes exhibited by many human patients. Finally, we show that AUTS2-l ablation in Calbindin 1-expressing cell lineages is sufficient to yield learning/memory deficits and hyperactivity with abnormal dentate gyrus granule cell maturation, but not other phenotypic effects. These data provide new clues to in vivo AUTS2-l functions and novel information relevant to genotype-phenotype correlations in the human AUTS2 region.

HLA-Homozygous iPSC-Derived Mesenchymal Stem Cells Rescue Rotenone-Induced Experimental Leber's Hereditary Optic Neuropathy-like Models In Vitro and In Vivo.

BACKGROUND: Mesenchymal stem cells (MSCs) hold promise for cell-based therapy, yet the sourcing, quality, and invasive methods of MSCs impede their mass production and quality control. Induced pluripotent stem cell (iPSC)-derived MSCs (iMSCs) can be infinitely expanded, providing advantages over conventional MSCs in terms of meeting unmet clinical demands. METHODS: The potential of MSC therapy for Leber's hereditary optic neuropathy (LHON) remains uncertain. In this study, we used HLA-homozygous induced pluripotent stem cells to generate iMSCs using a defined protocol, and we examined their therapeutic potential in rotenone-induced LHON-like models in vitro and in vivo. RESULTS: The iMSCs did not cause any tumorigenic incidence or inflammation-related lesions after intravitreal transplantation, and they remained viable for at least nine days in the mouse recipient's eyes. In addition, iMSCs exhibited significant efficacy in safeguarding retinal ganglion cells (RGCs) from rotenone-induced cytotoxicity in vitro, and they ameliorated CGL+IPL layer thinning and RGC loss in vivo. Optical coherence tomography (OCT) and an electroretinogram demonstrated that iMSCs not only prevented RGC loss and impairments to the retinal architecture, but they also improved retinal electrophysiology performance. CONCLUSION: The generation of iMSCs via the HLA homozygosity of iPSCs offers a compelling avenue for overcoming the current limitations of MSC-based therapies. The results underscore the potential of iMSCs when addressing retinal disorders, and they highlight their clinical significance, offering renewed hope for individuals affected by LHON and other inherited retinal conditions.

Retina-TransNet: A Gradient-Guided Few-Shot Retinal Vessel Segmentation Net.

Due to the high labor cost of physicians, it is difficult to collect a rich amount of manually-labeled medical images for developing learning-based computer-aided diagnosis (CADx) systems or segmentation algorithms. To tackle this issue, we reshape the image segmentation task as an image-to-image (I2I) translation problem and propose a retinal vascular segmentation network, which can achieve good cross-domain generalizability even with a small amount of training data. We devise primarily two components to facilitate this I2I-based segmentation method. The first is the constraints provided by the proposed gradient-vector-flow (GVF) loss, and, the second is a two-stage Unet (2Unet) generator with a skip connection. This configuration makes 2Unet's first-stage play a role similar to conventional Unet, but forces 2Unet's second stage to learn to be a refinement module. Extensive experiments show that by re-casting retinal vessel segmentation as an image-to-image translation problem, our I2I translator-based segmentation subnetwork achieves better cross-domain generalizability than existing segmentation methods. Our model, trained on one dataset, e.g., DRIVE, can produce segmentation results stably on datasets of other domains, e.g., CHASE-DB1, STARE, HRF, and DIARETDB1, even in low-shot circumstances.

Isolated loss of the AUTS2 long isoform, brain-wide or targeted to Calbindin -lineage cells, generates a specific suite of brain, behavioral and molecular pathologies.

Rearrangements within the AUTS2 region are associated with a rare syndromic disorder with intellectual disability, developmental delay and behavioral abnormalities as core features. In addition, smaller regional variants are linked to wide range of neuropsychiatric disorders, underscoring the gene's essential role in brain development. Like many essential neurodevelopmental genes, AUTS2 is large and complex, generating distinct long (AUTS2-l) and short (AUTS2-s) protein isoforms from alternative promoters. Although evidence suggests unique isoform functions, the contributions of each isoform to specific AUTS2- linked phenotypes have not been clearly resolved. Furthermore, Auts2 is widely expressed across the developing brain, but cell populations most central to disease presentation have not been determined. In this study, we focused on the specific roles of AUTS2-l in brain development, behavior, and postnatal brain gene expression, showing that brain-wide AUTS2-l ablation leads to specific subsets of the recessive pathologies associated with C-terminal mutations that disrupt both isoforms. We identify downstream genes that could explain expressed phenotypes including hundreds of putative direct AUTS2- l target genes. Furthermore, in contrast to C-terminal Auts2 mutations which lead to dominant hypoactivity, AUTS2-l loss-of-function is associated with dominant hyperactivity, a phenotype exhibited by many human patients. Finally, we show that AUTS2-l ablation in Calbindin 1 -expressing cell lineages is sufficient to yield learning/memory deficits and hyperactivity with abnormal dentate gyrus granule cell maturation, but not other phenotypic effects. These data provide new clues to in vivo AUTS2-l functions and novel information relevant to genotype-phenotype correlations in the human AUTS2 region.

Hepatic-Modulatory Effects of Chicken Liver Hydrolysate-Based Supplement on Autophagy Regulation against Liver Fibrogenesis.

Chicken-liver hydrolysates (CLHs) have been characterized as performing several biofunctions by our team. This study aimed to investigate if a CLH-based supplement (GBHP01TM) can ameliorate liver fibrogenesis induced by thioacetamide (TAA) treatment. Our results showed that the TAA treatment caused lower body weight gains and enlarged livers, as well as higher serum ALT, AST, and ALP levels (p < 0.05). This liver inflammatory and fibrotic evidence was ameliorated (p < 0.05) by supplementing with GBHP01TM; this partially resulted from its antioxidant abilities, including decreased TBARS values but increased TEAC levels, reduced GSH contents and catalase/GPx activities in the livers of TAA-treated rats (p < 0.05). Additionally, fewer nodules were observed in the appearance of the livers of TAA-treated rats after supplementing with GBHP01TM. Similarly, supplementing GBHP01TM decreased fibrotic scars and the fibrotic score in the livers of TAA-treated rats (p < 0.05). Moreover, the increased hepatic IL-6, IL-1ß, and TNF-α levels after TAA treatment were also alleviated by supplementing with GBHP01TM (p < 0.05). Meanwhile, GBHP01TM could decrease the ratio of LC3B II/LC3B I, but upregulated P62 and Rab7 in the livers of TAA-treated rats (p < 0.05). Taking these results together, the CLH-based supplement (GBHP01TM) can be characterized as a natural agent against liver fibrogenesis.

Galnt17 loss-of-function leads to developmental delay and abnormal coordination, activity, and social interactions with cerebellar vermis pathology.

GALNT17 encodes a N-acetylgalactosaminyltransferase (GalNAc-T) protein specifically involved in mucin-type O-linked glycosylation of target proteins, a process important for cell adhesion, cell signaling, neurotransmitter activity, neurite outgrowth, and neurite sensing. GALNT17, also known as WBSCR17, is located at the edge of the Williams-Beuren Syndrome (WBS) critical region and adjacent to the AUTS2 locus, genomic regions associated with neurodevelopmental phenotypes that are thought to be co-regulated. Although previous data have implicated Galnt17 in neurodevelopment, the in vivo functions of this gene have not been investigated. In this study, we have analyzed behavioral, brain pathology, and molecular phenotypes exhibited by Galnt17 knockout (Galnt17-/-) mice. We show that Galnt17-/- mutants exhibit developmental neuropathology within the cerebellar vermis, along with abnormal activity, coordination, and social interaction deficits. Transcriptomic and protein analysis revealed reductions in both mucin type O-glycosylation and heparan sulfate synthesis in the developing mutant cerebellum along with disruption of pathways central to neuron differentiation, axon pathfinding, and synaptic signaling, consistent with the mutant neuropathology. These brain and behavioral phenotypes and molecular data confirm a specific role for Galnt17 in brain development and suggest new clues to factors that could contribute to phenotypes in certain WBS and AUTS2 syndrome patients.

Impacts of Saharan Mineral Dust on Air-Sea Interaction over North Atlantic Ocean Using a Fully Coupled Regional Model.

This study examines the modifications of air-sea coupling processes by dust-radiation-cloud interactions over the North Atlantic Ocean using a high-resolution coupled atmosphere-wave-ocean-dust (AWOD) regional model. The dust-induced mechanisms that are responsible for changes of sea surface temperature (SST) and latent and sensible heat fluxes (LHF/SHF) are also examined. Two 3-month numerical experiments are conducted, and they differ only in the activation and deactivation of dust-radiation-cloud interactions. Model results show that the dust significantly reduces surface downward radiation fluxes (SDRF) over the ocean with the maximum change of 20-30 W m-2. Over the dust plume region, the dust effect creates a low-pressure anomaly and a cyclonic circulation anomaly, which drives a positive wind stress curl anomaly, thereby reducing sea surface height and mixed layer depth. However, the SST change by dust, ranging from -0.5 to 0.5 K, has a great spatial variation which differs from the dust plume shape. Dust cools SST around the West African coast, except under the maximum dust plume ridge, and extends westward asymmetrically along the northern and southern edges of the dust plume. Dust unexpectedly warms SST over a large area of the western tropical North Atlantic and north of the dust plume. These SST changes are controlled by different mechanisms. Unlike the SST change pattern, the LHF and SHF changes are mostly reduced underneath the dust plume region, though they are different in detail due to different dominant factors, and increased south of the dust plume over the tropic.

Evaluating the Impact of Assimilating Aerosol Optical Depth Observations on Dust Forecasts Over North Africa and the East Atlantic Using Different Data Assimilation Methods.

This study evaluates the impact of assimilating moderate resolution imaging spectroradiometer (MODIS) aerosol optical depth (AOD) data using different data assimilation (DA) methods on dust analyses and forecasts over North Africa and tropical North Atlantic. To do so, seven experiments are conducted using the Weather Research and Forecasting dust model and the Gridpoint Statistical Interpolation analysis system. Six of these experiments differ in whether or not AOD observations are assimilated and the DA method used, the latter of which includes the three-dimensional variational (3D-Var), ensemble square root filter (EnSRF), and hybrid methods. The seventh experiment, which allows us to assess the impact of assimilating deep blue AOD data, assimilates only dark target AOD data using the hybrid method. The assimilation of MODIS AOD data clearly improves AOD analyses and forecasts up to 48 hr in length. Results also show that assimilating deep blue data has a primarily positive effect on AOD analyses and forecasts over and downstream of the major North African source regions. Without assimilating deep blue data (assimilating dark target only), AOD assimilation only improves AOD forecasts for up to 30 hr. Of the three DA methods examined, the hybrid and EnSRF methods produce better AOD analyses and forecasts than the 3D-Var method does. Despite the clear benefit of AOD assimilation for AOD analyses and forecasts, the lack of information regarding the vertical distribution of aerosols in AOD data means that AOD assimilation has very little positive effect on analyzed or forecasted vertical profiles of backscatter.

Intra-auricular modification of facelift incision decreased the risk of Frey syndrome.

OBJECTIVE: Frey syndrome is a complication followed by parotidectomy which caused gustatory sweating and facial flush. There were several methods for the prevention of Frey syndrome, but most of them had no obvious effects. In this study, we compare the intra-auricular modification of facelift incision with the traditional lazy-S incision to see if it can decrease the risk of Frey syndrome. MATERIALS AND METHODS: This is a retrospective study. From 2003 to 2009, a total of 61 patients with benign parotid tumor who received parotidectomy at Hualien Tzu Chi Hospital and were followed at outpatient department for at least 5 years were enrolled. Patients were divided into two groups according to the type of incisions during operation: (1) Group M: intra-auricular modification of facelift incision or (2) Group S: traditional lazy-S incision. All patients received the partial thickness sternocleidomastoid muscle flap. Clinical data including age, gender, pathologic result, presentation of Frey syndrome, size of tumor, length of operation, blood loss from surgery, length of placement of drain, total amount of drainage, and length of stay were collected and analyzed. RESULTS: Sixty-one patients were enrolled. Eighteen patients were in Group M and forty-three were in Group S. There was no significant difference of age, gender, and size of tumor between the two groups. The pathologic results included parotitis, pleomorphic adenoma, Warthin's tumor, and others. No significant difference of pathologic results, blood loss from surgery, length of placement of drain, total amount of drainage, and length of stay between two groups was obtained. The length of operation was longer in Group M (P = 0.001) and the incidence of Frey syndrome was lower in Group M than Group S (P < 0.05). CONCLUSIONS: The use of intra-auricular modification of facelift incision can decrease the incidence of Frey syndrome.

A Mouse Mutation That Dysregulates Neighboring Galnt17 and Auts2 Genes Is Associated with Phenotypes Related to the Human AUTS2 Syndrome.

AUTS2 was originally discovered as the gene disrupted by a translocation in human twins with Autism spectrum disorder, intellectual disability, and epilepsy. Since that initial finding, AUTS2-linked mutations and variants have been associated with a very broad array of neuropsychiatric disorders, sugg esting that AUTS2 is required for fundamental steps of neurodevelopment. However, genotype-phenotype correlations in this region are complicated, because most mutations could also involve neighboring genes. Of particular interest is the nearest downstream neighbor of AUTS2, GALNT17, which encodes a brain-expressed N-acetylgalactosaminyltransferase of unknown brain function. Here we describe a mouse (Mus musculus) mutation, T(5G2;8A1)GSO (abbreviated 16Gso), a reciprocal translocation that breaks between Auts2 and Galnt17 and dysregulates both genes. Despite this complex regulatory effect, 16Gso homozygotes model certain human AUTS2-linked phenotypes very well. In addition to abnormalities in growth, craniofacial structure, learning and memory, and behavior, 16Gso homozygotes display distinct pathologies of the cerebellum and hippocampus that are similar to those associated with autism and other types of AUTS2-linked neurological disease. Analyzing mutant cerebellar and hippocampal transcriptomes to explain this pathology, we identified disturbances in pathways related to neuron and synapse maturation, neurotransmitter signaling, and cellular stress, suggesting possible cellular mechanisms. These pathways, coupled with the translocation's selective effects on Auts2 isoforms and coordinated dysregulation of Galnt17, suggest novel hypotheses regarding the etiology of the human "AUTS2 syndrome" and the wide array of neurodevelopmental disorders linked to variance in this genomic region.

Experimental generation and analysis of chaos-modulated pulses for pulsed chaos lidar applications based on gain-switched semiconductor lasers subject to optical feedback.

We experimentally generate and analyze chaos-modulated pulses for pulsed chaos lidar applications based on gain-switched semiconductor lasers subject to optical feedback. While conventional pulsed lidars emit repetitive short pulses without specificity making them vulnerable to interference and range ambiguity, chaos lidars possess the advantages of having no range ambiguity and being immune to interference and jamming, which are benefits of the aperiodic and uncorrelated waveforms we use. Compared to the cw chaos lidars originally proposed, the pulsed chaos lidars can have significantly higher peak power under the class-1 eye-safe regulation that is essential for long-range low-reflectivity target detection. We investigate the temporal, spectral, and cross-correlation characteristics of the modulated pulses obtained with different feedback strengths and modulation currents. Induced by the transient response and evolving with the delayed feedback, modulated pulses exhibiting periodic oscillations and complex dynamics such as chaos are observed. Under a weakly damped condition with large modulation current and moderate feedback strength, we successfully generate uncorrelated chaos-modulated pulses suitable for the pulsed chaos lidar applications. With the current configuration, for cross-correlations comparable to the benchmark of 0.19 set by the cross-correlation of the intensity fluctuation on the sole gain-switched pulses without feedback, uncorrelated waveforms with durations up to 218 ns in a 500 ns modulated pulse can be effectively utilized.

3D pulsed chaos lidar system.

We develop an unprecedented 3D pulsed chaos lidar system for potential intelligent machinery applications. Benefited from the random nature of the chaos, conventional CW chaos lidars already possess excellent anti-jamming and anti-interference capabilities and have no range ambiguity. In our system, we further employ self-homodyning and time gating to generate a pulsed homodyned chaos to boost the energy-utilization efficiency. Compared to the original chaos, we show that the pulsed homodyned chaos improves the detection SNR by more than 20 dB. With a sampling rate of just 1.25 GS/s that has a native sampling spacing of 12 cm, we successfully achieve millimeter-level accuracy and precision in ranging. Compared with two commercial lidars tested side-by-side, namely the pulsed Spectroscan and the random-modulation continuous-wave Lidar-lite, the pulsed chaos lidar that is in compliance with the class-1 eye-safe regulation shows significantly better precision and a much longer detection range up to 100 m. Moreover, by employing a 2-axis MEMS mirror for active laser scanning, we also demonstrate real-time 3D imaging with errors of less than 4 mm in depth.

Evaluation of the US Food and Drug Administration sentinel analysis tools in confirming previously observed drug-outcome associations: The case of clindamycin and Clostridium difficile infection.

PURPOSE: The Food and Drug Administration's Sentinel System developed parameterized, reusable analytic programs for evaluation of medical product safety. Research on outpatient antibiotic exposures, and Clostridium difficile infection (CDI) with non-user reference groups led us to expect a higher rate of CDI among outpatient clindamycin users vs penicillin users. We evaluated the ability of the Cohort Identification and Descriptive Analysis and Propensity Score Matching tools to identify a higher rate of CDI among clindamycin users. METHODS: We matched new users of outpatient dispensings of oral clindamycin or penicillin from 13 Data Partners 1:1 on propensity score and followed them for up to 60 days for development of CDI. We used Cox proportional hazards regression stratified by Data Partner and matched pair to compare CDI incidence. RESULTS: Propensity score models at 3 Data Partners had convergence warnings and a limited range of predicted values. We excluded these Data Partners despite adequate covariate balance after matching. From the 10 Data Partners where these models converged without warnings, we identified 807 919 new clindamycin users and 8 815 441 new penicillin users eligible for the analysis. The stratified analysis of 807 769 matched pairs included 840 events among clindamycin users and 290 among penicillin users (hazard ratio 2.90, 95% confidence interval 2.53, 3.31). CONCLUSIONS: This evaluation produced an expected result and identified several potential enhancements to the Propensity Score Matching tool. This study has important limitations. CDI risk may have been related to factors other than the inherent properties of the drugs, such as duration of use or subsequent exposures.

Multimode optical feedback dynamics in InAs/GaAs quantum dot lasers emitting exclusively on ground or excited states: transition from short- to long-delay regimes.

The optical feedback dynamics of two multimode InAs/GaAs quantum dot lasers emitting exclusively on sole ground or excited lasing states is investigated. The transition from long- to short-delay regimes is analyzed, while the boundaries associated to the birth of periodic and chaotic oscillations are unveiled to be a function of the external cavity length. The results show that depending on the initial lasing state, different routes to chaos are observed. These results are of importance for the development of isolator-free transmitters in short-reach networks.

Comparison of optical feedback dynamics of InAs/GaAs quantum-dot lasers emitting solely on ground or excited states.

We experimentally compare the dynamics of InAs/GaAs quantum dot lasers under optical feedback emitting exclusively on ground states (GSs) or excited states (ESs). By varying the feedback parameters and putting focus either on their short or long cavity regions, various periodic and chaotic oscillatory states are found. The GS laser is shown to be more resistant to feedback, benefiting from its strong relaxation oscillation damping. In contrast, the ES laser can easily be driven into complex dynamics. While the GS laser is of importance for the development of isolator-free transmitters, the ES laser is essential for applications taking advantages of chaos.

Acacetin from Traditionally Used Saussurea involucrata Kar. et Kir. Suppressed Adipogenesis in 3T3-L1 Adipocytes and Attenuated Lipid Accumulation in Obese Mice.

Acacetin, a flavone that can be isolated from the Saussurea involucrata plant, has anti-tumor and anti-inflammatory properties that ameliorate airway hyperresponsiveness in asthmatic mice. This study investigated whether acacetin has anti-adipogenic effects in 3T3-L1 adipocytes and whether it regulates the inflammatory response in adipocytes and macrophages. It also investigated whether acacetin ameliorates lipid accumulation in high-fat diet- (HFD) induced obese mice. Differentiated 3T3-L1 cells were treated with acacetin. The glycerol levels in the culture medium were measured, and the expression of proteins and genes involved in adipogenesis and lipolysis were assayed by Western blot and real-time PCR, respectively. Inflammatory cytokine signaling pathway activity was assessed in macrophages that were treated with acacetin and cultured with differentiated medium from 3T3-L1 cells. Intraperitoneal injections of acacetin were administered to HFD-induced obese mice twice a week for 10 weeks. Acacetin significantly increased the levels of glycerol in the culture medium and significantly inhibited lipid accumulation in adipocytes. Acacetin reduced the expression of adipogenesis-related transcription factors, including the expression of the CCAAT/enhancer-binding protein; it also increased sirtuin 1 expression and AMPK phosphorylation in adipocytes. In macrophages cultured with differentiated media from 3T3-L1 adipocytes, acacetin reduced the levels of inflammatory mediators and the activity of the mitogen-activated protein kinase and NF-κB pathways. In obese mice, acacetin reduced both body weight and visceral adipose tissue weight. These results demonstrate that acacetin inhibited adipogenesis in adipocytes and in obese mice. Acacetin also reduced the inflammatory response of macrophages that were stimulated with differentiated media from 3T3-L1 cells.

TALEN-Mediated Homologous Recombination Produces Site-Directed DNA Base Change and Herbicide-Resistant Rice.

Over the last decades, much endeavor has been made to advance genome editing technology due to its promising role in both basic and synthetic biology. The breakthrough has been made in recent years with the advent of sequence-specific endonucleases, especially zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeats (CRISPRs) guided nucleases (e.g., Cas9). In higher eukaryotic organisms, site-directed mutagenesis usually can be achieved through non-homologous end-joining (NHEJ) repair to the DNA double-strand breaks (DSBs) caused by the exogenously applied nucleases. However, site-specific gene replacement or genuine genome editing through homologous recombination (HR) repair to DSBs remains a challenge. As a proof of concept gene replacement through TALEN-based HR in rice (Oryza sativa), we successfully produced double point mutations in rice acetolactate synthase gene (OsALS) and generated herbicide resistant rice lines by using TALENs and donor DNA carrying the desired mutations. After ballistic delivery into rice calli of TALEN construct and donor DNA, nine HR events with different genotypes of OsALS were obtained in T0 generation at the efficiency of 1.4%-6.3% from three experiments. The HR-mediated gene edits were heritable to the progeny of T1 generation. The edited T1 plants were as morphologically normal as the control plants while displayed strong herbicide resistance. The results demonstrate the feasibility of TALEN-mediated genome editing in rice and provide useful information for further genome editing by other nuclease-based genome editing platforms.