Telocytes (TCs), a distinct interstitial cell population, are increasingly being reported in the male reproductive system. This study aimed to confirm TC presence in goat testes, to characterize their immunophenotype and age-related differences. Testes from one-, two-, and twelve-month-old goats were examined by using transmission electron microscopy (TEM) and double immunofluorescence (DIF) staining. TEM revealed that TCs were widely distributed among peritubular myoid cells (PMC) in testes from 12-month-old goats and exhibited reduced cell bodies and cytoplasm compared to TCs in juvenile goats. Their telopodes (TPs) displayed the characteristic bead-like structure of alternating podomers (Pms) and podoms (Pds). The telopodes were slender and curved, intertwining near collagen bundles and blood vessels and releasing abundant vesicles. DIF staining reliably identified goat testicular TCs as CD34+/vimentin+ cells with elongated processes and spindle- or triangular-shaped bodies. TCs in 12-month-old goats were significantly longer than those in juvenile goats and had more elongated nuclei. They also exhibited markedly more protrusions and were arranged in four or five parallel stromal layers. The DIF findings corroborated the TEM observations. This study provides evidence of age-related morphological and distributional changes in goat testicular TCs, establishing a foundation for understanding their role in maintaining the microenvironmental homeostasis of the testes.

Fluxapyroxad, a broad-spectrum and high-efficiency succinate dehydrogenase inhibitor (SDHI) fungicide, can control plant pathogens on many crops. However, no data about the fungicidal activity of fluxapyroxad against Stemphylium solani and its resistance risk are available. In this study, the mycelial growth inhibition method was used to determine the sensitivity of 99 S. solani strains to fluxapyroxad. The EC50 values ranged from 0.0183 to 0.3017 mu g/mL, with an average EC50 value of 0.0839 +/- 0.0051 mu g/mL. The results indicated that fluxapyroxad exhibited an excellent inhibitory efficacy against S. solani. Six resistant mutants were obtained by laboratory fungicide taming. The resistance ratio of each mutant was greater than 50 and the resistance of the mutants to fluxapyroxad was steadily inheritable through 10 successive culture transfers to fungicide-free PSA plates. We found that fluxapyroxad resistance not only affected mycelial growth in S. solani, but was also temperature dependent. Moreover, fluxapyroxad resistance caused markedly decreased virulence of S. solani on detached tomato leaves, indicating that the resistant mutants suffered a fitness penalty. Cross-resistance analysis showed that no cross-resistance was observed between fluxapyroxad and fludioxonil, iprodione, azoxystrobin or fluazinam, while a positive cross-resistance was observed between fluxapyroxad and pydiflumetofen. We concluded that the resistance of S. solani to fluxapyroxad was moderate risk. Therefore, fluxapyroxad is recommended as a potential candidate for controlling tomato gray leaf spot caused by S. solani.

The esophagus is a tubular organ which act as a passage for food from oral cavity to stomach. Telocytes (TCs) are a unique type of interstitial cell whose existence in many organs of various species still remains unknown. In the present study, we used transmission electron microscopy (TEM) and immunohistochemistry (CD34, Vimentin, PDGFR-α) to identify subepithelial TCs in the esophageal wall of chickens. TEM micrographs confirmed the presence of TCs in the lamina propria, tunica submucosa, and tunica muscularis muscular layer of the esophageal wall. A large population of TCs were observed just beneath the epithelial layer of the esophageal wall, and the TCs demonstrated structural heterogenicity, featuring various cell body shapes of cell bodies and telopodes (Tps) with podoms, podomeres, and dichotomous branching. Furthermore, a large number of extracellular vesicles were found to be associated with TCs/Tps. Cellular extensions from TCs were observed in close proximity to blood vessels, immune cells, and mucosal glands. In the submucosa, Tps and immune cells were in very close contact. Immunohistochemical results showed that there were CD34+ cells, vimentin+ cells, and PDGFR-α+ cells in the subepithelium, lamina propria, and mucosal glands of the chicken esophageal wall, which was consistent with the TEM results. Overall, our data confirmed the existence of TCs in the chicken esophagus and suggested that TCs might contribute to epithelial regeneration and tissue homeostasis.

Telocytes (TCs) are a new type of mesenchymal cells that have been discovered recently in many organs and tissues. However, studies of TCs in the avian reproductive system are still at the beginning. Chickens are one of the world's most popular domesticated animals, providing inexpensive but valuable proteins and nutrients from chickens and eggs to nourish the human bodies. Chickens have important scientific value; thus, understanding the reproductive system regulations seems to be important. The utero-vaginal junction is involved in the regulation of sperm storage. The sperm storage tube (SST) in the utero-vaginal junction stores sperm. The purpose of this study was to investigate the existence of TCs in the utero-vaginal junction of the chicken, and their structural relationships with the sperm storage tube and surrounding cell types. We studied the morphology, ultrastructure, and immune characterization of TCs. Methods The utero-vaginal junction of 4-month-old healthy adult chickens (n = 10) were used for Masson's staining, fluorescent in situ hybridization technique (FISH), and transmission electron microscopy (TEM) analysis. The results showed that TCs were present in the utero-vaginal junction. TCs appear as CD34 immunopositive and C-kit immunopositive. They were identified especially via small-body and long-protrusion telopodes (Tps) containing Podomers (Pm) and Podoms (Pd). The Tps were bent, folded, and intertwined with each other, sometimes in the shape of a labyrinth. The Tps were embedded between collagen fiber bundles, smooth muscle bundles, and around blood vessels and releasing vesicles. TCs surround these glands, forming heteromorphic cell connections with surrounding lymphocytes and plasma cells, smooth muscle cells, blood vessels, collagen fibers, and fibroblast-formed homotypic or allotypic connections in a complex three-dimensional network structure. This study provides a morphological basis for the possible role of TCs in regulating the utero-vaginal junction physiological role and in intercellular communication.

Abstract Telocyte (TC)—a new type of interstitial cell with long telopodes, can form cellular junctions with various tissues or cells to participate in the regulation of multitudes of physiological activities and diseases. This study aimed to characterize the morphology, molecular features, and potential functions of hormone regulation in Chinese soft-shelled turtle (Pelodiscus sinensis) testis TCs at different reproductive stages by histological evaluation, immunohistochemistry (IHC), immunofluorescence (IF), and transmission electron microscopy. During hibernation, TCs were widely distributed in the interstitial tissue. In contrast, during reproductive activity, TCs were noted to be in close proximity with peritubular myoid cells surrounding the seminiferous tubule. Moreover, formed cell–cell junctions were observed between TCs and PTMs. The results of IHC and IF showed that the immunophenotype of testicular TCs in hibernating Chinese soft-shelled turtles is CD34+Vimentin−, while the reproductive telopodes (Tps) show low expression of vimentin. The androgen receptor is expressed in Tps of TCs of testis during hibernation. Our results showed also that TCs in seasonal breeding animals regulate the activity of neighboring cells by releasing extracellular microvesicles (EXMVs), thus influencing the activity of spermatogenesis and steroidogenesis. Consideration of our novel and interesting results indicate that the whole area warrants further research.

The dermal microvascular unit (DMU) is a perivascular functional unit in the dermis. It is composed of microvascular and capillary lymphatics surrounded by immune cells. In this study, jet needle-free injection system was used to injected biocompatible carbon nanoparticles into the cervical skin of domestic pigs (Sus scrofa domestica) and assessed the morphological distribution of DMUs by hematoxylin erythrosine staining, immunohistochemistry (IHC), and transmission electron microscopy (TEM), and TEM was also used to observe the ultrastructural changes of DMUs after jet needle-free injection. Following our study, we identified DMUs in the dermis stratum papillare and similar structures in the dermis stratum reticulare, but the aggregation of CD68+ and CD1a+ cells in the dermis stratum papillare of DMUs by IHC confirmed that DMUs act as reservoirs of dermal immune cells, while similar structures in the dermis stratum reticulare should not be considered as DMUs. Ultrastructure of DMUs was revealed by TEM. Marvelous changes were found following xenobiotics attack, including the rearrangement of endothelial cells and pericytes, and the reactivity of immune cells. Novel interstitial cell telocyte (TC) was also identified around the microvasculature, which may have been previously known as the veil cell. Our results successfully identified the distribution of DMUs in the skin of domestic pigs, which might act as reservoirs of immune cells in the skin and play a role in immune surveillance and immune defense.

We have previously reported that bovine papillomavirus type 1 (BPV1) can replicate its genome and produces infectious virus-like particles in short-term BPV1 virion-infected Sacharomyces cerevisiae (Zhao and Frazer, 2002). Here, we report viral RNA transcription and L1 capsid protein expression in long-term BPV1 virion-infected S. cerevisiae culture. Northern blot hybridization showed that viral RNA was detected in long-term BPV1-infected S. cerevisiae cultures (82-108 days). The levels of the viral RNA transcription varied significantly over the long time period, which showed active transcription at an early stage (Day 3 to Day 16), weak transcription at a middle stage (Day 23 to Day 45) and stable transcription at the late stage of culture (Day 55 to Day 82/85/95). Three major BPV1 transcripts of 4.3, 2.6 and 1.8 Kb were identified, with 4.3 Kb a minor transcript and the 1.8 Kb the most prominent transcript compared with the 2.6 Kb species. Immunoblotting showed that L1 capsid protein was expressed, with its variable amounts corresponding to the levels of RNA transcription over the time period. 35S-methionine/cysteine labeling and immunoprecipitation proved that the detected L1 protein was newly synthesized in BPV1-infected S. cerevisiae cultures. 33.3-54.2% of the cell colonies expressed L1 protein. Thus, the S. cerevisiae system, as a promising model, may be used not only for the study of virus like particle formation of BPV1 in vitro, but also for further functional analysis of individual viral genes in BPV1 life cycle. Keywords: BPV1; viral RNA transcription; expression of L1 capsid protein; virion-infected Saccharomyces cerevisiae.

A preferable approach of a combination of a multiwavelength fusion HPLC fingerprint and chemometrics for the quality control of Xiaoer Chiqiao Qingre granules (XCQG) was established in this study. A single-wavelength HPLC fingerprint was performed to identify 18 peaks as common peaks in the beginning, and 12 of them were recognized by HPLC-Q/TOF-MS. To overcome the limitation of the single-wavelength HPLC fingerprint, a three-wavelength (230 nm, 250 nm, and 330 nm) fusion fingerprint was established for a more thorough quality assessment. Six main active ingredients (geniposide, paeoniflorin, forsythin, forsythoside A, baicalin, and wogonoside) were selected as chemical markers for simultaneous quantitative analysis, while the results indicated that the content of other five ingredients except forsythoside A presented comparatively stable. Chemometrics including hierarchical cluster analysis (HCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were performed to evaluate the homogeneity and heterogeneity of sixteen batches of XCQG. The results of the multiwavelength fingerprint were clearly classified into two clusters by HCA, whereas the single-wavelength fingerprint showed no distinct difference between them. OPLS-DA was further employed to prove that the above six main active ingredients made great contributions to clustering. In summary, this integrated analysis provided a better promoted and more comprehensive method to control the quality of XCQG.

Fresh blueberries were processed into three powder forms through freeze drying, namely juice powders containing 8% whey protein isolate (WPI), fruit powders and pomace powders. The water binding behaviors of these blueberry powders were studied through thermodynamic sorption analysis, quantification of water plasticizing effect and characterization of particle properties. The GAB model was successful in fitting the water sorption isotherms of blueberry powders at 20, 35 and 50°C. Thermodynamic studies revealed that the sorption processes of all the samples were enthalpy-driven and only the sorption process of blueberry juice powders containing WPI was spontaneous (ΔG<0). Meanwhile, the highest absolute integral entropy values for blueberry juice powders containing 8% WPI, fruit powders and pomace powders were obtained at the moisture contents of 0.011, 0.013 and 0.024kgH2O/kg dried matter, respectively. Furthermore, the plasticizing effect of water on blueberry pomace powders was the weakest, since its k value of Gordon-Taylor model was lower than that of other samples. These results implied that blueberry pomace powders had the weakest affinity with water molecules. Thus, blueberry juice powders containing 8% WPI and fruit powders were suggested to be stored at a lower relative humidity than pomace powders. Besides, it was temporarily found that the difference in water binding capacity among blueberry powders was associated with the distribution of functional groups derived from sugars and acids, as well as the particle size. All the results can be a reference for drying, packaging, storage and distribution of blueberry powders in practical applications.

The aim of this article was to study the influence of Saccharomyces cerevisiae strains on the chemical composition and sensory properties of mulberry wines. For this purpose, three Saccharomyces cerevisiae strains, YJM 681 (isolated from raspberry), ySR 127 (commercial yeast) and Y1 (isolated from kefir fermented milk) were used for the alcoholic fermentation of mulberry juice. Yeast strains had deep influences on oenological, phenolic, amino acid, volatile and sensory profiles of resulting mulberry wines. Specifically, the beverages fermented with YJM 681 were characterised by relatively high volatile acidity and high contents of total flavonols, amino acids and some phenolic acids, including protocatechuic acid, p-hydroxybenzoic acid, caffeic acid and veratric acid. Samples fermented with ySR 127 possessed higher cyanidin-3-O-glucoside content, whereas, total amino acid content was markedly lower than that of other samples. Furthermore, mulberry wines fermented with Y1 showed the lowest volatile acidity and the lowest contents of total anthocyanins and tartaric esters. The amounts of major aromatic compounds, including 1-propanol, 2-methyl-1-propanol and ethyl acetate inside were also smaller than that of samples fermented with YJM 681 and ySR 127. Besides, the sensory results revealed that mulberry wines fermented with Y1 exhibited the lowest intensity of fruity aroma, while samples fermented with ySR 127 had the strongest sour and sweet tastes and aftertaste. These findings provide fundamental knowledge about the influence of yeast strains on the quality of mulberry wine.

ABSTRACTGrasses exhibit diurnal variation in chemical composition and epiphytic microorganisms, which may influence the natural fermentation attributes of silage. The objective was to determine the effect of diurnal changes on chemical composition and epiphytic microorganisms and these effects on fermentation quality of unwilted Italian ryegrass (Lolium multiflorum Lam.) silage. Primary growth of Italian ryegrass was harvested in the heading stage at 0800 (AM), 1300 (M) and 1800 h (PM) on three sunny days. Each cutting time was assayed in triplicate in laboratory silos and sampled after 30 d (cutting Day 1) or serial days until 30 d (other cutting days) to analyze the fermentation attributes and microbial counts. Bacterial community was determined by denaturing gradient gel electrophoresis (DGGE) on pre‐ and post‐ensiled forage samples, which were harvested on cutting Day 2. The results showed that fermentation instability for the silage made with M‐cut forage, which had fewer lactic acid bacteria (LAB) species in the early phase of ensiling and where Clostridium sp. was present but no Lactobacillus plantarum at the end of ensiling, resulted in higher (P < 0.05) pH value, NH3–N, and butyric acid for 30‐d fermented silages than those made with AM‐ and PM‐cut forages. The concentrations of dry matter (DM) and lactic acid were higher (P < 0.05) and the silage pH and NH3–N were lower (P < 0.05) for silage made with PM‐cut forage as compared to those made with AM‐cut forages. Italian ryegrass cut at 1800 h was associated with good silage fermentation attributes.