Polarized Th2 cells attenuate high-fat-diet induced obesity through the suppression of lipogenesis.

Immune cells, such as macrophages, B cells, neutrophils and T cell subsets, have been implicated in the context of obesity. However, the specific role of Th2 cells in adipose tissue function has remained elusive. Eight-week-old male CD3ε─/─ mice were randomly divided into two groups (≥ 5 mice per group): one received intravenous injection of Th2 cells isolated from LATY136F mice, while the other receiving PBS as a control. Both of groups were subjected to a high-fat diet (HFD). The adoptive transfer of polarized Th2 cells led to a significant reduction in obesity following a HFD. This reduction was accompanied by improvements in hepatic steatosis, glucose intolerance, and insulin resistance. Mechanistically, Th2 cell treatment promoted oxidative phosphorylation of adipocytes, thereby contributing to a reduction of lipid droplet accumulation. These findings suggest that Th2 cell therapy represents a novel approach for treating diet-induced obesity and other diseases involving lipid droplet accumulation disorders.

Comparative genomics provides insights into the aquatic adaptations of mammals.

The ancestors of marine mammals once roamed the land and independently committed to an aquatic lifestyle. These macroevolutionary transitions have intrigued scientists for centuries. Here, we generated high-quality genome assemblies of 17 marine mammals (11 cetaceans and six pinnipeds), including eight assemblies at the chromosome level. Incorporating previously published data, we reconstructed the marine mammal phylogeny and population histories and identified numerous idiosyncratic and convergent genomic variations that possibly contributed to the transition from land to water in marine mammal lineages. Genes associated with the formation of blubber (NFIA), vascular development (SEMA3E), and heat production by brown adipose tissue (UCP1) had unique changes that may contribute to marine mammal thermoregulation. We also observed many lineage-specific changes in the marine mammals, including genes associated with deep diving and navigation. Our study advances understanding of the timing, pattern, and molecular changes associated with the evolution of mammalian lineages adapting to aquatic life.

The transcription factor GhWRKY70 from gossypium hirsutum enhances resistance to verticillium wilt via the jasmonic acid pathway.

BACKGROUND: The WRKY transcription factors play significant roles in plant growth, development, and defense responses. However, in cotton, the molecular mechanism of most WRKY proteins and their involvement in Verticillium wilt tolerance are not well understood. RESULTS: GhWRKY70 is greatly up-regulated in cotton by Verticillium dahliae. Subcellular localization suggests that GhWRKY70 is only located in the nucleus. Transcriptional activation of GhWRKY70 further demonstrates that GhWRKY70 function as a transcriptional activator. Transgenic Arabidopsis plants overexpressing GhWRKY70 exhibited better growth performance and higher lignin content, antioxidant enzyme activities and jasmonic acid (JA) levels than wild-type plants after infection with V. dahliae. In addition, the transgenic Arabidopsis resulted in an enhanced expression level of AtAOS1, a gene related to JA synthesis, further leading to a higher JA accumulation compared to the wild type. However, the disease index (DI) values of the VIGS-treated cotton plants with TRV:WRKY70 were also significantly higher than those of the VIGS-treated cotton plants with TRV:00. The chlorophyll and lignin contents of TRV:WRKY70 plants were significantly lower than those of TRV:00 plants. GhAOS1 expression and JA abundance in TRV:WRKY70 plants were decreased. The GhWRKY70 protein was confirmed to bind to the W-box element in the promoter region of GhAOS by yeast one-hybrid assay and transient expression. CONCLUSION: These results indicate that the GhWRKY70 transcription factor is a positive regulator in Verticillium wilt tolerance of cotton, and may promote the production of JA via regulation of GhAOS1 expression.

Glycolate oxidase gene family identification and functional analyses in cotton resistance to Verticillium wilt.

Glycolate oxidase (GOX) plays an important role in the regulation of photorespiration and photosynthesis in plants. However, as one of the main enzymes to produce the second messenger hydrogen peroxide (H2O2), its functions in response to pathogens are still poorly understood. In this study, we carried out genome-wide identification, and 14 GOX genes were identified in Gossypium hirsutum. These GOX genes are located on 10 chromosomes and divided into hydroxyacid-oxidases (HAOX) and GOX groups. After infection with Verticillium dahliae Kleb., six GOX gene expression levels were changed. Moreover, H2O2, salicylic acid (SA), and the content and activity of GOX increased in cotton. GhHAOX2-D-silenced plants showed more wilting than control plants after infection with V. dahliae. Additionally, H2O2 accumulation and SA content were reduced in GhHAOX2-D-silenced plants. The expression levels of GhPAL, GhPAD4, and GhPR1 and the lignin content of the silenced plants were significantly lower than those of the control plants. These results indicate that GhHAOX2-D is a positive regulator of Verticillium wilt tolerance in cotton and may promote H2O2 accumulation via the synergistic effects of GOX genes and SA. Collectively, GOX genes play an important role in cotton resistance to Verticillium wilt.

Echolocation clicks of free-ranging Indo-Pacific finless porpoises (Neophocaena phocaenoides) in Hainan waters.

The echolocation clicks of free-ranging Indo-Pacific finless porpoises (IPFPs, Neophocaena phocaenoides) have been rarely studied in the wild. This paper aims at describing the echolocation-click characteristics of IPFPs and examining whether IPFPs adapt their sonar system to the habitats in Hainan waters, China. The echolocation clicks were recorded using a 13 elements star-shaped array of hydrophones. A total of 65 on-axis clicks were identified and analyzed. IPFPs use echolocation clicks with a source level (SL) of 158 ± 9 dB re: 1 µPa peak-peak, mean peak, and centroid frequency of 134 ± 3 kHz, -3 dB bandwidth of 14 ± 2 kHz and produce at inter-click intervals of 104 ± 51 ms. The results relative to other porpoises show that finless porpoises in Hainan waters produce clicks with moderate SLs and high peak frequency. These results could be useful in detecting the presence and estimating the density of IPFPs during passive acoustic monitoring in the study area and serve to shed light on the interpopulation variation of click characteristics of finless porpoises as well.

Idelalisib inhibits experimental proliferative vitroretinopathy.

Proliferative vitreoretinopathy (PVR) is a fibrotic eye disease that develops after rhegmatogenous retinal detachment surgery and open-globe traumatic injury. Idelalisib is a specific inhibitor of phosphoinositide 3-kinase (PI3K) δ. While PI3Kδ is primarily expressed in leukocytes, its expression is also considerably high in retinal pigment epithelial (RPE) cells, which play a crucial part in the PVR pathogenesis. Herein we show that GeoMx Digital Spatial Profiling uncovered strong expression of fibronectin in RPE cells within epiretinal membranes from patients with PVR, and that idelalisib (10 µM) inhibited Akt activation, fibronectin expression and collagen gel contraction induced by transforming growth factor (TGF)-ß2 in human RPE cells. Furthermore, we discovered that idelalisib at a vitreal concentration of 10 µM, a non-toxic dose to the retina, prevented experimental PVR induced by intravitreally injected RPE cells in rabbits assessed by experienced ophthalmologists using an indirect ophthalmoscope plus a + 30 D fundus lens, electroretinography, optical coherence tomography and histological analysis. These data suggested idelalisib could be harnessed for preventing patients from PVR.

Low-cost polymer-film spiral inertial microfluidic device for label-free separation of malignant tumor cells.

We developed a low-cost polymer-film spiral inertial microfluidic device for the effective size-dependent separation of malignant tumor cells. The device was fabricated in polymer films by rapid laser cutting and chemical bonding. After fabricating the prototype device, the separation performance of our device was evaluated using particles and cells. The effects of operational flow rate, cell diameter, and cell concentration on the separation performance were explored. Our device successfully separated tumor cells from polydisperse white blood cells according to their different migration modes and lateral positions. Then, the separation of rare cells was carried out using the high-concentration lysed blood spiked with 200 tumor cells. Experimental results showed that 83.90% of the tumor cells could be recovered, while 99.87% of white blood cells could be removed. We successfully employed our device for processing clinical pleural effusion samples from patients with advanced metastatic breast cancer. Malignant tumor cells with an average purity of 2.37% could be effectively enriched, improving downstream diagnostic accuracy. Our device offers the advantages of label-free operation, low cost, and fast fabrication, thus being a potential tool for effective cell separation.

A robust method for protein depletion based on gene editing.

The technology of clustered regularly interspaced short palindromic repeats (CRISPR)-associated nuclease Cas9 (CRISPR-Cas9) is a powerful system for protein depletion resulting from insertions and deletions following Cas9 cleavage of genome at specific site in vitro and in vivo. We herein present a relatively standard protocol for protein depletion in a step-by-step procedure, including guide RNA designation and vector construction, lentivirus production, cell selection, and experimentally validate the function of targeted protein. We exemplified this approach by editing PDGFRß in human epithelial cells, and expected that this simplified and detailed protocol will be more broadly applied on specific genes to aid understanding gene functions.

AAV-mediated in vivo genome editing in vascular endothelial cells.

In vivo genome editing meets numerous challenges including efficiency and safety. Here we describe an efficient in vivo genome editing method of delivering CRISPR-Cas9 into vascular endothelial cells with adeno-associated viruses (AAVs). In this system, expression of SpCas9 is driven by a specific endothelial promoter of intercellular adhesion molecule 2 (pICAM2) to restrict this foreign enzyme in vascular endothelial cells, which can be efficiently infected by AAV1. We exemplify this approach by editing VEGFR2 in retinal vascular endothelial cells in a mouse model of oxygen-induced retinopathy, and expect that this simplified protocol can be expanded to other researches on editing endothelial genome in vivo.

Chalcomoracin prevents vitreous-induced activation of AKT and migration of retinal pigment epithelial cells.

Retinal pigment epithelial (RPE) cells are the major cell type in the epi- or sub-retinal membranes in the pathogenesis of proliferative vitreoretinopathy (PVR), which is a blinding fibrotic eye disease and still short of effective medicine. The purpose of this study is to demonstrate whether Chalocomoracin (CMR), a novel purified compound from fungus-infected mulberry leaves, is able to inhibit vitreous-induced signalling events and cellular responses intrinsic to PVR. Our studies have revealed that the CMR IC50 for ARPE-19 cells is 35.5 µmol/L at 72 hours, and that 5 µmol/L CMR inhibits vitreous-induced Akt activation and p53 suppression; in addition, we have discovered that this chemical effectively blocks vitreous-stimulated proliferation, migration and contraction of ARPE-19 cells, suggesting that CMR is a promising PVR prophylactic.

Numerical-modeling-based investigation of sound transmission and reception in the short-finned pilot whale (Globicephala macrorhynchus).

The sound-transmission, beam-formation, and sound-reception processes of a short-finned pilot whale (Globicephala macrorhynchus) were investigated using computed tomography (CT) scanning and numerical simulation. The results showed that sound propagations in the forehead were modulated by the upper jaw, air components, and soft tissues, which attributed to the beam formation in the external acoustic field. These structures owned different acoustic impedance and formed a multiphasic sound transmission system that can modulate sounds into a beam. The reception pathways composed of the solid mandible and acoustic fats in the lower head conducted sounds into the tympano-periotic complex. In the simulations, sounds were emitted in the forehead transmission system and propagated into water to interrogate a steel cylinder. The resulting echoes can be interpreted from multiple perspectives, including amplitude, waveform, and spectrum, to obtain the acoustic cues of the steel cylinder. By taking the short-finned pilot whale as an example, this study provides meaningful information to further deepen our understanding of biosonar system operations, and may expand sound-reception theory in odontocetes.

Mannan-binding lectin attenuates acetaminophen-induced hepatotoxicity by regulating CYP2E1 expression via ROS-dependent JNK/SP1 pathway.

Mannan-binding lectin (MBL) acts as a soluble pattern recognition molecule in the innate immune system, which is primarily produced by the liver. MBL deficiency occurs with high frequency in the population and is reported to be associated with susceptibility to several liver diseases. In the present study, we investigated the pathophysiological role of MBL in acetaminophen (APAP)-induced hepatotoxicity. After APAP treatment, MBL-deficient (MBL-/- ) mice had significantly higher mortality and aggravated hepatic necrosis as well as elevated serum lactate dehydrogenase and alanine aminotransferase levels compared to control mice. The enhanced hepatotoxicity in MBL-/- mice was associated with increased concentration of APAP toxic metabolisms. Furthermore, we demonstrated here that genetic ablation of MBL resulted in excessive reactive oxygen species (ROS) production and enhanced c-Jun N-terminal kinase (JNK) activation, leading to up-regulated specificity protein 1 (SP1) nuclear expression, thus promoted CYP2E1 hepatic expression and consequently exacerbated APAP-induced liver injury in mice. Importantly, we have validated that MBL protected against APAP toxicity in human HepaRG cells in vitro with the same mechanism. Our study revealed an unexpected function of MBL in drug metabolism, thus providing new insight into the drug-induced liver injury in patients with MBL deficiency.

Comparative analysis of two brine shrimps revealed differential expression pattern and functional characterization of CK2α under bacterial stimulation from different geographical distribution.

Understanding how the brine shrimp responds to different geographical populations can provide novel insights on response to bacterial stimulation. In the paper, Artemia sinica from lower altitudes and Artemia parthenogenetica from higher altitudes of the Tibetan Plateau, were used to illustrate different defense against bacteria mechanisms that these organisms used to adapt to different geographical environments. Protein kinase CK2 is a serine/threonine kinase with a multitude of protein substrates. It is a ubiquitous enzyme essential for the viability of eukaryotic cells, where its functions in a variety of cellular processes, including cell cycle progression, apoptosis, transcription, and viral infection. The gene encodes the same mRNA sequence in A. sinica and A. parthenogenetica, named AsCK2α and ApCK2α, respectively. The open reading frame was obtained, a 1047-bp sequence encoding a predicted protein of 349 amino acids. To systematically analyze the expression of AsCK2α and ApCK2α during embryonic development and bacterial challenge, real-time PCR, Western blotting and immunohistochemistry were performed. The results showed that AsCK2α was higher than ApCK2α at different developmental stages. Under bacterial challenge, the expression of ApCK2α was significantly higher than AsCK2α. Protein localization analysis showed that AsCK2α and ApCK2α were mainly distributed in the head and chest. Our research revealed that CK2α plays a vital role in the growth, development and bacterial stimulation of the brine shrimp.

Gut Microbiota and Immune Responses.

The gut microbiota consists of a dynamic multispecies community living within a particular niche in a mutual synergy with the host organism. Recent findings have revealed roles for the gut microbiota in the modulation of host immunity and the development and progression of immune-mediated diseases. Besides, growing evidence supports the concept that some metabolites mainly originated from gut microbiota are linked to the immune regulation implicated in systemic inflammatory and autoimmune disorders. In this chapter, we describe the recent advances in our understanding of how host-microbiota interactions shape the immune system, how they affect the pathogenesis of immune-associated diseases and the impact of these mechanisms in the efficacy of disease therapy.

Influence of acoustic habitat variation on Indo-Pacific humpback dolphin (Sousa chinensis) in shallow waters of Hainan Island, China.

The Indo-Pacific humpback dolphin (IPHD, Sousa chinensis) is a coastal species inhabiting tropical and warm-temperate waters. The presence of this vulnerable dolphin was recently discovered in shallow waters southwest of Hainan Island, China. The influence of the acoustic habitat on the distribution and behavior of IPHD was investigated using an array of passive acoustic platforms (n = 6) that spanned more than 100 km of coastline during a 75-day monitoring period. Its presence was assessed within 19 215 five-min recordings by classifying echolocation clicks using machine learning techniques. Spectrogram analysis was applied to further investigate the acoustic behavior of IPHD and to identify other prominent sound sources. The variation in the ambient noise levels was also measured to describe the spatiotemporal patterns of the acoustic habitat among the different sampling sites. Social and feeding sounds of IPHD (whistles and click-series of pulsed sounds) were identified together with other biological sources (finless porpoise, soniferous fishes, and snapping shrimps) and anthropogenic activities (ship noise, explosions, and sonars). Distribution, acoustic behavior, and habitat use of this nearshore dolphin species were strongly influenced by the abundance of soniferous fishes, and under similar conditions, the species was more acoustically active in locations with lower noise levels.

Echolocation signals of free-ranging pantropical spotted dolphins (Stenella attenuata) in the South China Sea.

Echolocation signals of free-ranging pantropical spotted dolphins (Stenella attenuata) in the western Pacific Ocean have not been studied much. This paper aims to describe the characteristics of echolocation signals of S. attenuata in the northern South China Sea. A six-arm star array with 13 hydrophones was used and a total of 131 on-axis clicks were identified to analyze the acoustic features of the echolocation signals of dolphins. The mean center frequency was 89 ± 13 kHz, with mean peak-to-peak sound source levels of 190 ± 6 dB re: 1 µPa @ 1 m. The mean -3 dB bandwidth and root-mean-square bandwidth were 62 ± 15 kHz and 26 ± 3 kHz, respectively, with mean -10 dB duration of 18 ± 4 µs and root-mean-square duration of 6 ± 2 µs. The results showed that click parameters of S. attenuata in the northern South China Sea are different from those of clicks of the species in Hawaii waters. The differences in click parameters may be due to both behavioral context and/or environmental adaptation of S. attenuata in different habitats.

Whistles emitted by Indo-Pacific humpback dolphins (Sousa chinensis) in Zhanjiang waters, China.

Whistles emitted by Indo-Pacific humpback dolphins in Zhanjiang waters, China, were collected by using autonomous acoustic recorders. A total of 529 whistles with clear contours and signal-to-noise ratio higher than 10 dB were extracted for analysis. The fundamental frequencies and durations of analyzed whistles were in ranges of 1785-21 675 Hz and 30-1973 ms, respectively. Six tonal types were identified: constant, downsweep, upsweep, concave, convex, and sine whistles. Constant type was the most dominant tonal type, accounting for 32.51% of all whistles, followed by sine type, accounting for 19.66% of all whistles. This paper examined 17 whistle parameters, which showed significant differences among the six tonal types. Whistles without inflections, gaps, and stairs accounted for 62.6%, 80.6%, and 68.6% of all whistles, respectively. Significant intraspecific differences in all duration and frequency parameters of dolphin whistles were found between this study and the study in Malaysia. Except for start frequency, maximum frequency and the number of harmonics, all whistle parameters showed significant differences between this study and the study conducted in Sanniang Bay, China. The intraspecific differences in vocalizations for this species may be related to macro-geographic and/or environmental variations among waters, suggesting a potential geographic isolation among populations of Indo-Pacific humpback dolphins.

Physiological responses of three soybean species (Glycine soja, G. gracilis, and G. max cv. Melrose) to salinity stress.

Understanding the mechanism for salt tolerance in wild soybean (Glycine soja) can help researchers improve that trait in cultivated soybean lines. We analyzed the effects of excess NaCl on the growth, physiology, and ion distribution in three soybean species: wild G. soja (W8), semi-wild G. gracilis (SW18), and the cultivated salt-sensitive G. max (cv. Melrose). These comparisons revealed that, under salt stress, shoot and root lengths and biomass (either shoot or root dry weights) were significantly higher for the W8 genotype than for the other two. Most of the morphological parameters for roots from the W8 plants were also increased, including total length, specific root length, and surface area. However, the average root diameter for W8 was significantly lower than that of either SW18 or 'Melrose' soybeans. In response to salinity, photosynthesis was suppressed to a greater extent in 'Melrose' than in W8. The relatively higher tolerance shown by W8 plants was also associated with lower levels of malondialdehyde, hydrogen peroxide, and relative electrical conductivity, but higher activities by superoxide dismutase and peroxidase, as well as more free proline and glycine betaine. In addition, the W8 plants contained less Na+ and Cl-, but more K+, and they had a higher K+/Na+ ratio in their leaves and roots when compared with either SW18 or 'Melrose' plants. Therefore, the W8 genotype performs better in terms of seedling growth, photosynthetic characteristics, and physiological indexes. These findings provide guidance for developing new soybean cultivars with improved tolerance to salt stress. Our data also contribute to the knowledge base for plant salt tolerance as a tool for increasing the yields of other crops in high-salinity soils.

Broadband ship noise and its potential impacts on Indo-Pacific humpback dolphins: Implications for conservation and management.

Ship noise pollution has raised considerable concerns among regulatory agencies and cetacean researchers worldwide. There is an urgent need to quantify ship noise in coastal areas and assess its potential biological impacts. In this study, underwater broadband noise from commercial ships in a critical habitat of Indo-Pacific humpback dolphins was recorded and analyzed. Data analysis indicated that the ship noise caused by the investigated commercial ships with an average length of 134 ± 81 m, traveling at 18.8 ± 2.5 km/h [mean ± standard deviation (SD), n = 21] comprises mid-to-high components with frequencies approaching and exceeding 100 kHz, and the ship noise could be sensed auditorily by Indo-Pacific humpback dolphins within most of their sensitive frequency range. The contributions of ship noise to ambient noise were highest in two third-octave bands with center frequencies of 8 and 50 kHz, which are within the sensitive hearing range of Indo-Pacific humpback dolphins and overlap the frequency of sounds that are biologically significant to the dolphins. It is estimated that ship noise in these third-octave bands can be auditorily sensed by and potentially affect the dolphins within 2290 ± 1172 m and 848 ± 358 m (mean ± SD, n = 21), respectively.

Acoustic occurrence detection of a newly recorded Indo-Pacific humpback dolphin population in waters southwest of Hainan Island, China.

In 2014, Indo-Pacific humpback dolphins were recorded for the first time in waters southwest of Hainan Island, China. In this paper, the temporal occurrence of Indo-Pacific humpback dolphins in this region was detected by stationary passive acoustic monitoring. During the 130-day observation period (from January to July 2016), 1969 click trains produced by Indo-Pacific humpback dolphins were identified, and 262 ten-minute recording bins contained echolocation click trains of dolphins, of which 70.9% were at night and 29.1% were during the day. A diurnal rhythm with a nighttime peak in acoustic detections was found. Passive acoustic detections indicated that the Indo-Pacific humpback dolphins frequently occurred in this area and were detected mainly at night. This information may be relevant to conservation efforts for these dolphins in the near future.