An alternative σ factor σL sl regulates lincomycin production in Streptomyces lincolnensis.

Lincomycin produced by Streptomyces lincolnensis is a critical antibacterial antibiotic in the clinical. To further understand the regulatory mechanism of lincomycin biosynthesis, we identified an alternative σ factor, σL sl , in Streptomyces lincolnensis NRRL 2936. Deletion of sigLsl resulted in an increase in cell growth but a decrease in lincomycin production. σL sl boosted lincomycin biosynthesis by directly stimulating the transcription of four genes (lmbD, lmbV, lmrC, and lmbU) within the lincomycin biosynthetic lmb gene cluster. Besides, σL sl participated in lincomycin biosynthesis by directly stimulating the transcription of mshC, a gene responsible for MSH synthesis. In conclusion, our findings demonstrated that σL sl plays a direct regulatory role in lincomycin biosynthesis. This study extends the understanding of molecular mechanisms of lincomycin biosynthetic regulation.

LSLOpt: An open-source implementation of the step-length controlled LSL-BFGS algorithm.

Numerical optimization is a common technique in various areas of computational chemistry, molecular modeling and drug design. It is a key element of 3D techniques, for example, the optimization of protein-ligand poses and small-molecule conformers. Here, often the BFGS algorithm or variants thereof are used. However, the BFGS algorithm tends to make unreasonable large changes to the optimized system under certain circumstances. This behavior has been known for a long time and different solutions have been suggested. Recently, we have analyzed the optimization behavior of our novel JAMDA scoring function in detail and proposed the limited step length (LSL)-BFGS algorithm as a new solution to the problem of excessively large steps during optimization. The LSL-BFGS algorithm allows to control the step sizes during optimization. Its unique feature is the inclusion of arbitrary domain knowledge into the selection of the step sizes. Here, we introduce the open-source LSLOpt C++ library that implements this LSL-BFGS algorithm and demonstrate its usage.

Generation of a novel mouse strain with conditional, cell-type specific, expression of DND1.

Dead-End 1 (DND1) encodes an RNA binding protein critical for viable primordial germ cells in vertebrates. When introduced into cancer cell lines, DND1 suppresses cell proliferation and enhances apoptosis. However, the molecular function of mammalian wild-type DND1 has mostly been studied in cell lines and not verified in the organism. To facilitate study of wild-type DND1 function in mammalian systems, we generated a novel transgenic mouse line, LSL-FM-DND1 flox/+ , which conditionally expresses genetically engineered, FLAG-tagged and myc-tagged DND1 in a cell type-specific manner. We report that FLAG-myc-DND1 is indeed expressed in specific tissues of the mouse when LSL-FM-DND1 flox/+ is combined with mouse strains expressing Cre-recombinase. LSL-FM-DND1 flox/+ mice are fertile with no overt health effects. We expressed FLAG-myc-DND1 in the pancreas and found that chronic, ectopic expression of FLAG-myc-DND1 led to increase in fasting glucose levels in older mice. Thus, this novel LSL-FM-DND1 flox/+ mouse strain will facilitate studies on the biological and molecular function of wild-type DND1.

Molecular cloning, phylogenetic analysis, and expression patterns of LATERAL SUPPRESSOR-LIKE and REGULATOR OF AXILLARY MERISTEM FORMATION-LIKE genes in sunflower (Helianthus annuus L.).

The wild sunflower (Helianthus annuus) plants develop a highly branched form with numerous small flowering heads. The origin of a no branched sunflower, producing a single large head, has been a key event in the domestication process of this species. The interaction between hormonal factors and several genes organizes the initiation and outgrowth of axillary meristems (AMs). From sunflower, we have isolated two genes putatively involved in this process, LATERAL SUPPRESSOR (LS)-LIKE (Ha-LSL) and REGULATOR OF AXILLARY MERISTEM FORMATION (ROX)-LIKE (Ha-ROXL), encoding for a GRAS and a bHLH transcription factor (TF), respectively. Typical amino acid residues and phylogenetic analyses suggest that Ha-LSL and Ha-ROXL are the orthologs of the branching regulator LS and ROX/LAX1, involved in the growth habit of both dicot and monocot species. qRT-PCR analyses revealed a high accumulation of Ha-LSL transcripts in roots, vegetative shoots, and inflorescence shoots. By contrast, in internodal stems and young leaves, a lower amount of Ha-LSL transcripts was observed. A comparison of transcription patterns between Ha-LSL and Ha-ROXL revealed some analogies but also remarkable differences; in fact, the gene Ha-ROXL displayed a low expression level in all organs analyzed. In situ hybridization (ISH) analysis showed that Ha-ROXL transcription was strongly restricted to a small domain within the boundary zone separating the shoot apical meristem (SAM) and the leaf primordia and in restricted regions of the inflorescence meristem, beforehand the separation of floral bracts from disc flower primordia. These results suggested that Ha-ROXL may be involved to establish a cell niche for the initiation of AMs as well as flower primordia. The accumulation of Ha-LSL transcripts was not restricted to the boundary zones in vegetative and inflorescence shoots, but the mRNA activity was expanded in other cellular domains of primary shoot apical meristem as well as AMs. In addition, Ha-LSL transcript accumulation was also detected in leaves and floral primordia at early stages of development. These results were corroborated by qRT-PCR analyses that evidenced high levels of Ha-LSL transcripts in very young leaves and disc flowers, suggesting a role of Ha-LSL for the early outgrowth of lateral primordia.

How does visible light flicker impact laying hen pullet behavior, fear, and stress levels?

Many characteristics of artificial light have been evaluated; however, light-flicker frequency (F) has not been assessed extensively in poultry. Pullets (1,344 per strain [S]; Lohmann Brown-Lite [LB] and LSL-Lite [LW]) were placed into 8 light-tight rooms, containing 6 floor pens (15 pen replicates per F × S for 30 and 250 Hz; 18 pen replicates per F x S for 90 Hz), and assigned 1 of 3 F treatments (30, 90, 250 Hz). The experiment took place over 2 trials (blocks). To evaluate long-term effects of F during rearing, birds were followed through the hen phase. Data were analyzed using Proc Mixed (SAS 9.4). Differences were considered significant when P ≤ 0.05, and behaviors are expressed as percentage of time. Pullets reared under 30 Hz spent more time performing nutritive behaviors (P < 0.01) and as "unidentified" (P = 0.02) than other treatments. Active behavior demonstrated an age x F interaction, with pullets being more active at 16 wk, regardless of F (P < 0.01). Comfort behaviors were higher at 16 wk compared to other ages, regardless of F (P < 0.01). Exploratory behaviors were lowest at 4 wk in pullets under 30 Hz (P < 0.01). Aggressive behaviors (12 wk) were higher in pullets reared under 250 Hz than those under 90 Hz (P < 0.01). Comb score was unaffected by F (P = 0.79) and all birds scored had a full plumage. Heterophil-to-lymphocyte ratio was unaffected by F at 7 or 15 wk (P = 0.85 and P = 0.54, respectively). In trial 1, pullets reared under 90 Hz had higher corticosterone concentrations than those reared under 250 Hz (P = 0.02) and trial 2 there were no effects of F (P = 0.97). For novel object test, LW pullets reared under 90 Hz had a higher latency to peck than LW pullets under 30 Hz or 250 Hz (P = 0.03). Hen behavior (wk 39) and fear tests (36 wk; novel object test (P = 0.86) and tonic immobility (P = 0.37)) were unaffected by F. Overall, minimal effects of F were seen on pullet and hen behavior and stress.

Do flickering LED lights reduce productivity of layer pullets and hens?

Most characteristics of artificial light sources are well studied, however light-flicker frequency (F) has been overlooked. The purpose of this study was to determine the effect of F on performance of Lohmann LSL-Lite (LW) pullets and Lohmann Brown-Lite (LB) pullets. In addition, pullets were followed through to the laying phase to evaluate long-term effects of F during rearing on productivity. Two trials were conducted with 3 F (30, 90, or 250 Hz) treatments. LW and LB pullets (n = 2,688 per strain [S]) were randomly assigned to floor pens within 8 light-tight rooms (15 pen replicates per F × S for 30 and 250 Hz; 18 pen replicates per F × S for 90 Hz). At 16 wk, pullets were transferred to conventional layer cages, with no flicker treatment applied. Pullet data collected included BW, feed disappearance, flock uniformity, and overall mortality. Hen data collected included BW, feed intake (feed efficiency calculated), mortality, egg production, and egg quality. Data were analyzed using Proc Mixed (SAS 9.4) and differences were considered significant when P ≤ 0.05. Frequency did not affect pullet uniformity or feed disappearance (0-8 wk and 0-16 wk). Pullets reared under 30 Hz had higher mortality (caused by "other") than those reared under 250 Hz. Lohmann Brown-Lite pullets reared under 30 Hz had the highest feed disappearance. Overall mortality was higher for LW pullets reared under 30 Hz compared to LB reared under 30 Hz or 250 Hz. Lohmann Brown-Lite hens reared under 30 Hz were heavier at the beginning of the hen phase (17 wk), however differences related to F were not seen at 40 or 48 wk. Hen day production (%) was higher for hens reared under 30 compared to 90 Hz (P = 0.03), however no other egg parameters were affected by F. Hen feed efficiency and mortality were unaffected by F. These results indicate minor effects of F, during either the pullet or hen phases. The data also suggest that S (LW vs. LB) may affect response to F.

Atorvastatin inhibits pancreatic carcinogenesis and increases survival in LSL-KrasG12D-LSL-Trp53R172H-Pdx1-Cre mice.

There are several studies supporting the role of HMG-CoA reductase inhibitors such as atorvastatin against carcinogenesis, in which inhibiting the generation of prenyl intermediates involved in protein prenylation plays the crucial role. Mutation of Kras gene is the most common genetic alteration in pancreatic cancer and the Ras protein requires prenylation for its membrane localization and activity. In the present study, the effectiveness of atorvastatin against pancreatic carcinogenesis and its effect on protein prenylation were determined using the LSL-KrasG12D-LSL-Trp53R172H-Pdx1-Cre mouse model (called Pankras/p53 mice). Five-week-old Pankras/p53 mice were fed either an AIN93M diet or a diet supplemented with 100 ppm atorvastatin. Kaplan-Meier survival analysis with Log-Rank test revealed a significant increase in survival in mice fed 100 ppm atorvastatin (171.9 ± 6.2 d) compared to the control mice (144.9 ± 8.4 d, P < 0.05). Histologic and immunohistochemical analysis showed that atorvastatin treatment resulted in a significant reduction in tumor volume and Ki-67-labeled cell proliferation. Mechanistic studies on primary pancreatic tumors and the cultured murine pancreatic carcinoma cells revealed that atorvastatin inhibited prenylation in several key proteins, including Kras protein and its activities, and similar effect was observed in pancreatic carcinoma cells treated with farnesyltransferase inhibitor R115777. Microarray assay on the global gene expression profile demonstrated that a total of 132 genes were significantly modulated by atorvastatin; and Waf1p21, cyp51A1, and soluble epoxide hydrolase were crucial atorvastatin-targeted genes which involve in inflammation and carcinogenesis. This study indicates that atorvastatin has the potential to serve as a chemopreventive agent against pancreatic carcinogenesis.

Effect of Dietary Supplementation of Zinc Nanoparticles Prepared by Different Green Methods on Egg Production, Egg Quality, Bone Mineralization, and Antioxidant Capacity in Caged Layers.

The present study was planned to evaluate the effect of dietary zinc-oxide (ZnO) nanoparticles synthesized by different plant extracts on egg production, egg quality, bone mineralization, and antioxidant capacity in caged layers. Nanoparticles of ZnO were synthesized by using extracts of Allium sativum (AS), Aloe vera (AV), Curcuma longa (CL), and Zingiber officinale (ZO). Different sources of nano ZnO (AS, AV, CL, and ZO) with varying levels (35, 70, or 105 ppm) were tested on 288 caged LSL layers of 25 weeks of age. Each diet was offered to 4 replicates of 6 birds each level and the duration of trial was 8 weeks. Daily egg production, feed consumption, and fortnightly egg quality parameters were recorded. Egg quality parameters (egg weight, egg mass, shape index, yolk index, albumen index, Haugh unit score, specific gravity, and eggshell thickness) were determined fortnightly by taking 2 eggs from each replicate randomly. Antioxidant capacity and bone mineralization were determined at the end of the trial. Results showed that the nano ZnO preparations were not effective (P < 0.05) on laying performance but additional levels (70 ppm) improved egg production, feed conversion ratio, egg mass, Haugh unit score, and antioxidant capacity of chickens. An interaction was found among nanoparticles prepared by Allium sativum and Zingiber officianale extracts with 70 ppm level regarding total antioxidant capacity and egg production (P > 0.05). Interaction among source and level was not found regarding feed intake, feed conversion ratio, egg quality, bone characteristics, and concentration of Zn. Results of the present study suggest that nano ZnO sources may not be a factor that affects performance, but level affects the birds' physiology. Thus, it is concluded that nano ZnO with 70 ppm concentration is sufficient to optimize the laying performance.

Molecular Interaction Mechanism and Preservative Effect of Lactone Sophorolipid and Lactoferrin/ß-Lactoglobulin Systems.

Multispectral and molecular docking methods were used to study the interaction mode and mechanism of two important components of whey proteins, lactoferrin (LF) and ß-lactoglobulin (ß-LG), and of a lactone sophorolipid (LSL) mixed system. The preservation effect of the mixed system on milk was also studied and compared. The results showed that the quenching mechanism of LSL on both ß-LG and LF was static, but that the non-covalent complexes formed were the result of the different interacting forces: hydrogen bonds and the van der Waals force for the LSL-ß-LG system, and electrostatic force for the LSL-LF system. The binding constants of LSL-ß-LG and LSL-LF were all relatively small, and the interaction of LSL with ß-LG was stronger than its interaction with LF. After adding ß-LG, LF, or the mixed system with LSL to the milk, the stability of milk emulsion was effectively improved in all cases, while the preservative ability was effectively enhanced only by the addition of LF or LSL-LF. These results provide supportive data and a theoretical basis for enhancing the production of dairy products and other byproducts.

One-step generation of mice with gene editing by Tol2 transposon-dependent gRNA delivery.

Conditional knockout mice are valuable tools for examining the functions of targeted genes in a time- and space-specific manner. Here, we generated gene-edited mice by using the Tol2 transposon to introduce guide RNA (gRNA) into fertilized eggs obtained by crossing LSL (loxP-stop-loxP)-CRISPR-associated 9 (Cas9) mice, which express Cas9 in a Cre-dependent manner, with CAG-CreER mice. Transposase mRNA and plasmid DNA, which contained a gRNA sequence for the gene encoding tyrosinase flanked by the transposase recognition sequence, were injected together into fertilized eggs. As a result, the transcribed gRNA cleaved the target genome in a Cas9-dependent manner. Using this method, it is possible to generate conditional genome-edited mice more easily in a shorter period of time.

Micro-straw: An efficient cryopreservation carrier for rare human spermatozoa.

BACKGROUND: Many cryopreservation carriers have been introduced to freeze rare human spermatozoa, however, these carriers relative attributes and comparative effectivenesses have not yet been systematically studied. OBJECTIVES: Is the micro-straw cryopreservation carrier more effective for cryopreserving rare human spermatozoa compared with the Cryoplus and a new micro-straw (LSL straw) carriers? MATERIALS AND METHODS: This study involves 93 samples from healthy sperm donors and 40 samples from patients diagnosed with oligospermia, asthenospermia, oligoasthenospermia, or obstructive azoospermia. We determined the optimal freeze-thaw protocol for the Micro-straw carrier. The post-thaw survival rate, normal sperm morphology, acrosome integrity, and DNA fragmentation for Micro-straw, Cryoplus, and LSL carriers were then determined. Finally, we verified the effects of freezing using these carriers by comparing the qualities of post-thaw spermatozoa from patients. RESULTS: The highest total motility (TM) and progressive motility (PR) survival rates were obtained by placing the Micro-straw at 1 cm above the LN2 surface for 70 s during freezing and in a 42°C water bath for 40 s during thawing. No differences were observed in the PR survival rate, acrosome integrity, and DNA fragmentation of the post-thaw spermatozoa from the three carriers. However, the normal morphology rate of spermatozoa frozen using the Micro-straw carrier was higher than for the Cryoplus carrier (p < 0.05), and the TM survival rate of spermatozoa frozen with the Micro-straw was higher than that for the LSL carrier (p < 0.01). In verification tests, there were no significant differences in the quality of post-thaw spermatozoa cryopreserved using these carriers for both rare spermatozoa or epididymal sperm. DISCUSSION AND CONCLUSION: Micro-straw, Cryoplus, and LSL carriers are all efficient means of freezing rare human spermatozoa. However, the Micro-straw carrier is more economical, safe, and user-friendly.

LSL1 controls cell death and grain production by stabilizing chloroplast in rice.

Lesion mutants can be valuable tools to reveal the interactions between genetic factors and environmental signals and to improve grain production. Here we identified a rice (Oryza sativa) mutant, lesion spotleaf1 (lsl1), which produces necrotic leaf lesions throughout its life cycle. LSL1 encodes a protein of unknown function and belongs to a grass-specific clade. The lesion phenotype of the lsl1 mutant was sharply induced by shading, and its detached leaves incubated in 6-benzylamino purine similarly formed lesions in the dark. In addition, the lsl1 mutant exhibited reactive oxygen species accumulation and cell death. The terminal deoxynucleotidyl transferase dUTP nick end-labeling (TUNEL) and comet assays revealed that the lsl1 mutant contained severe DNA damage, resulting in reduced grain yield and quality. RNA sequencing, gene expression, and protein activity analyses indicate that LSL1 is required for chloroplast function. Furthermore, LSL1 interacts with PsaD and PAP10 to form a regulatory module that functions in chlorophyll synthesis and chloroplast development to maintain redox balance. Our results reveal that LSL1 maintains chloroplast structure, redox homeostasis, and DNA stability, and plays important roles in the interaction between genetic factors and environmental signals and in regulating grain size and quality.

The Effect of Light Intensity, Strain, and Age on the Behavior, Jumping Frequency and Success, and Welfare of Egg-Strain Pullets Reared in Perchery Systems.

The effects of light intensity (L) are not well studied in pullets. Our research objective was to study the effect of L on navigational success, behavior, and welfare of two pullet strains (S). In two repeated trials, a 3 × 2 × 4 factorial arrangement tested three L (10, 30, 50 lux) and two S (Lohmann Brown-Lite (LB), LSL-Lite (LW)) at four ages. One thousand eight hundred pullets/S (0-16 wk) were randomly assigned to floor pens within light-tight rooms (three pens/S/room, four rooms/L) containing four parallel perches and a ramp. Data collection included jumping frequency and success (24h continuous sampling), novel object tests (fear), heterophil to lymphocyte (H/L) ratios (stress), and behavior (instantaneous scan sampling) during photoperiods. L did not affect injurious behavior, fear, or H/L. Pullets reared at 50 lux spent more time preening than at 10 lux. Pullets reared at 10 lux spent more time wall pecking than at 50 lux. Time spent standing and preening and total number and accuracy of jumping increased with age. Pullets reared at 30 lux had higher jumping frequency than at 10 lux; accuracy was not affected. LW jumped more than LB, but with similar success. LB spent more time exploring and scored higher in the fear and stress assessments, suggesting S differences.

The effect of light intensity on the body weight, keel bone quality, tibia bone strength, and mortality of brown and white feathered egg-strain pullets reared in perchery systems.

The development of the musculoskeletal system is influenced by bird activity, which can be impacted by light intensity (L). The objective of this study was to determine the effect of L on the growth and bone health of Lohmann Brown-Lite (LB) and Lohmann LSL-Lite (LW) pullets. Three L treatments (10, 30 or 50 lux, provided by white LED lights) were used in a Randomized Complete Block Design in 2 repeated trials. LB and LW (n = 1,800 per strain [S]) were randomly assigned to floor pens (50 pullets per pen; 12 pen replicates per L × S) within 6 light-tight rooms from 0 to 16 wk. Each pen contained 4 parallel perches and a ramp. Data collected include cumulative mortality, BW at 0, 8, and 16 wk, and uniformity, keel bone damage (KBD; deviations, fractures), breast muscle weight, and tibiae bone strength at 16 wk. Tibiae bone resistance to mechanical stress was assessed using a three-point-bending test. The effect of L, S, and their interactions were analyzed using Proc Mixed (SAS 9.4) and differences were considered significant when P < 0.05. L did not affect BW, KBD, or mortality. An interaction between L and S was observed for bone stress (bone strength relative to bone size), however, in general, LW pullets had greater resistance to bone stress (peak noted at 30 lux) than LB (peak at 50 lux). LB pullets were heavier than LW at 8 and 16 wk. There were no S differences on KBD from palpated or dissected keel bones. LB pullets had higher breast muscle weight and heavier tibiae than LW, however relative to BW, LW had a higher percentage of breast muscle and a longer and thicker tibiae than LB. LW had higher mortality during the first wk but there was no relationship to L. Conclusively, the results suggest that L, within a range of 10 to 50 lux, does not affect pullet BW or KBD, however S may affect both parameters, as well as bone strength.

Microarray Analysis Revealed Inflammatory Transcriptomic Changes after LSL60101 Treatment in 5XFAD Mice Model.

I2-IR have been found dysregulated in patients with neurodegenerative diseases, such as Alzheimer's disease (AD), in which the importance of neuroinflammation in the establishment and maintenance of cognitive decline is well-documented. To research the implication of I2-IR in neuroinflammatory pathways altered in AD, we determined the expression profile of genes associated with inflammation in the 5XFAD model treated with LSL60101, a well-established I2-IR ligand. Thus, we performed a qPCR array containing 84 inflammation-related genes. Hierarchical clustering analysis revealed three gene clusters, suggesting that treatment with LSL60101 affects the gene expression associated with inflammation in the 5XFAD model. Furthermore, we evaluated the functions of the three clusters; thereby performing a pathway enrichment analysis using the GO database. As we expected, clusters 2 and 3 showed alterations in the inflammatory response, chemotaxis and the chemokine-mediated signaling pathway, among others. To validate previous results from the gene profiling analysis, the expression levels of a representative subset of mRNAs were selected according to the intensity of the observed changes and their biological relevance. Interestingly, changes induced by LSL60101 in the 5XFAD model were validated for several genes. These results suggest that treatment with LSL60101 in the 5XFAD model reverses the inflammatory process during the development of AD.

Retrospective study on the incidence of pseudoexfoliatio lentis and related complications in a cohort of patients from the Island of Ischia: medico-legal and ergophthalmology considerations.

The population of the island has lived for centuries almost isolated from the rest of the continental population. In the seventh century BC, it was the first Greek colony in the southern Italy and was colonized by the Eubei, then it was the turn of the Roman rule. In the Middle Ages, the island was the victim of many raids of the Barbary pirates. Only in the last decades of the 800, did the island begin to open to tourism. However, it had a strong setback with the terrible earthquake of Casamicciola, in 1883. Tourism had gradually resumed in the second post-war period. These episodes have contributed to bringing people extraneous to the autochthonous community to the island. As in most of the coastal populations of the Mediterranean basin, there is a very high incidence of pseudoexfoliation lentis in the population of Ischia, which is a syndrome that often complicates in a challenging way the surgical intervention for the removal of cataracts.

Black raspberries suppress pancreatic cancer through modulation of NKp46+, CD8+, and CD11b+ immune cells.

Pancreatic ductal adenocarcinoma (PDAC) is a fatal disease with a low survival rate (9%). Epidemiologic studies show that healthy dietary patterns enriched of fruits and vegetables lower the risk of PDAC. We previously showed that supplementing black raspberries (BRBs) to patients with colorectal cancer increased tumor-infiltrating NK cells and their cytotoxicity. We aimed to determine whether BRBs combat PDAC by modulating cancer immunity. NOD.SCID mice lacking T and B cells were injected with human Panc-1-Luc cells orthotopically, and immunocompetent Kras LSL.G12D/+ -Trp53 LSL.R172H/+ -Pdx-1-Cre mice were fed BRBs. Peripheral blood mononuclear cells (PBMCs) from PDAC patients were treated with butyrate, a microbial metabolite of BRBs. The absence of T and B cells did not dampen BRBs' anti-tumor effects in the NOD.SCID mice. In the Kras LSL.G12D/+ -Trp53 LSL.R172H/+ -Pdx-1-Cre mice, BRBs significantly prolonged survival (189 days versus 154 days). In both models, BRBs decreased tumor-infiltrating CD11b+ cells and the expression of IL-1ß, sEH, and Ki67. BRBs also increased tumor-infiltrating NKp46+ cells and the expression of CD107a, a functional marker of cytolytic NK and CD8+ T cells. In Kras LSL.G12D/+ -Trp53 LSL.R172H/+ -Pdx-1-Cre mice, tumor infiltration of CD8+ T cells was increased by BRBs. Further using the PBMCs from PDAC patients, we show that butyrate decreased the population of myeloid-derived suppressor cells (MDSCs). Butyrate also reversed CD11b+ cell-mediated suppression on CD8+ T cells. Interestingly, there is a negative association between MDSC changes and patients' survival, suggesting that the more decrease in MDSC population induced by butyrate treatment, the longer the patient had survived. Our study suggests the immune-modulating potentials of BRBs in PDAC.

Capacity Coaching: A Focused Ethnographic Evaluation in Clinical Practice.

OBJECTIVE: To qualitatively evaluate the implementation of Capacity Coaching, an intervention to address the work patients must undertake to manage their conditions, implemented as a quality improvement pilot in 1 of 2 implementing US Department of Veterans Affairs medical centers. PARTICIPANTS AND METHODS: Two Veterans Affairs medical centers in the Midwest sought to implement Capacity Coaching as a quality improvement pilot in their Patient-Aligned Care Teams for 6 months (April 1, 2017, through October 31, 2017). Following the pilot, we conducted a focused ethnographic evaluation (on-site data collection, January 2-4, 2018), including interviews, a focus group, and observations with staff at one site to assess the implementation of capacity coaching. Data were analyzed inductively and findings were cross-referenced with implementation theory. RESULTS: We found that implementation was feasible and achieved changes that were aligned with reducing patient work and increasing capacity. We found that the key facilitators for the implementation of this program were in participants making sense of the intervention (coherence) and working collectively to enact the program (collective action). The main challenges for the program were in planning the work of implementation and enrolling a diverse coalition of staff to expand referrals to the program (cognitive participation) and in evaluating the impact of the program on outcomes that upper leadership was interested in (reflexive monitoring). CONCLUSION: Implementation of Capacity Coaching is feasible in clinical practice and may be a promising intervention for the care of chronic conditions. Further research should focus on testing capacity coaching using these lessons learned.

Neuron-Specific Genome Modification in the Adult Rat Brain Using CRISPR-Cas9 Transgenic Rats.

Historically, the rat has been the preferred animal model for behavioral studies. Limitations in genome modification have, however, caused a lag in their use compared to the bevy of available transgenic mice. Here, we have developed several transgenic tools, including viral vectors and transgenic rats, for targeted genome modification in specific adult rat neurons using CRISPR-Cas9 technology. Starting from wild-type rats, knockout of tyrosine hydroxylase was achieved with adeno-associated viral (AAV) vectors expressing Cas9 or guide RNAs (gRNAs). We subsequently created an AAV vector for Cre-dependent gRNA expression as well as three new transgenic rat lines to specifically target CRISPR-Cas9 components to dopaminergic neurons. One rat represents the first knockin rat model made by germline gene targeting in spermatogonial stem cells. The rats described herein serve as a versatile platform for making cell-specific and sequence-specific genome modifications in the adult brain and potentially other Cre-expressing tissues of the rat.

Mouse-Derived Allografts: A Complementary Model to the KPC Mice on Researching Pancreatic Cancer In Vivo.

Pancreatic ductal adenocarcinoma (PDAC) is one of the most malignant cancers and has an extremely undesirable prognosis because little is known about the initiation and progression mechanisms of pancreatic cancer. The lack of an appropriate research model may have hindered this process. Using LSL-Kras G12D/+ ; Trp53 fl/+ ; Pdx1-Cre (KPC) mice and the tumor tissue fragment transplantation technique, we constructed the mouse-derived subcutaneous/orthotopic allograft tumor models (MDAs-ST/OT). H&E staining, Masson staining and immunohistochemical staining were adopted to describe the histopathology and biomarkers of the MDAs and the recruitment of immune cells. The intervention of gemcitabine was applied to measure the chemotherapeutic response of MDAs tumors. MDAs could mimic the pathological histology and the high proliferation characteristics of PDAC. Indeed, the fibrosis, epithelial-mesenchyme transition (EMT) and invasion/metastasis related markers of MDAs were similar to those observed in pancreatic cancer. Further, the recruitment of immune cells in PDAC was precisely simulated by MDAs. In addition, gemcitabine suppressed the tumor growth of MDAs-ST significantly. MDAs are an effective model for investigating the progression and treatment of pancreatic cancer.