Comprehensive transcriptional profiling and functional study of lymphangiogenic growth factor in placentome of early-stage pregnant water buffalo

The aim of the present study was to evaluate the expression and localization of lymphangiogenic factors (VEGF-C and VEGF-D), their receptor (VEGFR3) and lymphatic endothelial marker (LYVE1) in buffalo placenta during early pregnancy [EP], and to investigate the functional role of lymphangiogenic growth factors in placental lymphangiogenesis. The mRNA and protein expression of VEGF-C, VEGF-D, their receptor VEGFR3 and LYVE1 showed significant expression in EP1 (29–42 days) and EP2 stages (51–82 days) both in caruncle (maternal part) and cotyledon (foetal part) of the buffalo placenta. Immunoreactivity of VEGF-C, VEGF-D and LYVE1 was observed around the endometrial gland, in lymphatics and trophoblast cells, whereas VEGFR3 mainly localized in lymphatics of the caruncle and cotyledons. Cultured trophoblast cells were treated with VEGF-C/VEGF-D (50, 100 and 150 ng/ml) and combined doses of VEGF-C and VEGF-D (150 ng/ml) each for different time durations (24, 48 and 72 h). The mRNA expression of LYVE1 and PCNA was significantly (p < .001) upregulated with VEGF-C and VEGF-D and combined treatment (@150 ng/ml), as well as significantly downregulating Caspase-3 at 48 and 72 h. Thus, the present study provides evidence that lymphangiogenic factors are expressed in buffalo placental compartments and they may play a significant role in the regulation of placental function in water buffaloes.     

Hydrogen cyanide production by soil bacteria: Biological control of pests and promotion of plant growth in sustainable agriculture

Currently, plant diseases and insect infestations are mainly controlled by the extraneous application of pesticides. Unfortunately, the indiscriminate use of such agrochemicals can cause ecological and environmental problems, as well as human health hazards. To obviate the potential pollution arising from the application of agrochemicals, biological control of soilborne pathogens or insect pests using antagonistic microorganisms may be employed. Certain soil bacteria, algae, fungi, plants and insects possess the unique ability to produce hydrogen cyanide(HCN), which plays an important role in the biotic interactions of those organisms. In particular, cyanogenic bacteria have been found to inhibit the growth of various pathogenic fungi, weeds, insects, termites and nematodes. Thus, the use of HCN-producing bacteria as biopesticides offers an ecofriendly approach for sustainable agriculture. The enzyme, HCN synthase,involved in the synthesis of HCN, is encoded by the hcnABC gene cluster. The biosynthetic regulation of HCN, antibiotics and fluorescent insecticidal toxins through the conserved global regulatory GacS/GacA system is elaborated in this review, including approaches that may optimize cyanogenesis for enhanced pest control. In addition, the effects of bacterially synthesized HCN on the production of indole acetic acid, antibiotics and fluorescent insecticidal toxins, 1-aminocyclopropane-1-carboxylate deaminase utilization and phosphate solubilization may result in the stimulation of plant growth. A more detailed understanding of HCN biosynthesis and regulation may help to elaborate the precise role of this compound in biotic interactions and sustainable agriculture.     

Bovine toll-like receptor 9: a comparative analysis of molecular structure, function and expression

Non-methylated CpG motifs, present in viral and bacterial DNA, are one of many pathogen-associated molecular patterns (PAMP) recognized by the mammalian innate immune system. Recognition of this PAMP occurs through a specific interaction with toll-like receptor 9 (TLR9) and this interaction can induce cytokine responses that influence both innate and adaptive immune responses. Previous investigations determined that both the flanking sequences in synthetic CpG oligodeoxynucleotides (CpG ODN) and the cellular pattern of TLR9 expression can influence species-specific responses to CpG ODN. Therefore, the structure, function and cellular distribution of bovine TLR9 were compared with what is known for mice and human. Analysis of the bovine TLR9 gene revealed greater sequence homology between cattle and humans than cattle and mice Similar CpG motifs induced optimal activation of both human and bovine leukocytes and these motifs were distinct from those which activated mouse leukocytes. Functional analyses with CpG ODN stimulated bovine blood leukocytes revealed that class A CpG ODN were more potent inducers of interferon-alpha (IFN-alpha) than class B CpG ODN. Furthermore, magnetic activated cell sorting of bovine blood leukocyte subpopulations implicated dendritic cells but not monocytes in the regulation of CpG ODN-induced IFN secretion. Thus, the cellular pattern of CpG ODN-induced responses in cattle shared many similarities with human leukocytes. Collectively, these analyses revealed substantial conservation of TLR9 structure and TLR9 function in blood leukocytes of humans, cattle and other domestic species.     

Quantitative analysis of pro-inflammatory cytokine mRNA expression in Theileria annulata-infected cell lines derived from resistant and susceptible cattle

The pathogenic mechanisms involved in tropical theileriosis, caused by the tick-borne protozoan parasite Theileria annulata, are unclear. Pathology is associated with the schizont stage of the parasite, which resides within bovine macrophages. Breed-specific differences in pathology have been observed in cattle, several Bos indicus breeds are relatively resistant to tropical theileriosis whilst Bos taurus cattle are highly susceptible. Infected cells express pro-inflammatory cytokines and it has been hypothesized that these cytokines play a major role in the pathology of the disease. Therefore, using quantitative RT-PCR we investigated the expression of the key candidates, interleukin 1 beta (IL-1beta), IL-6 and tumour necrosis factor alpha (TNF-alpha), in T. annulata low passage infected cell lines derived ex vivo from experimental infection of resistant and susceptible cattle. mRNA for each cytokine was detected in all cell lines investigated at levels higher than those observed in resting monocytes. However, the analyses did not identify any breed-specific differences. Therefore, these results are not consistent with the hypothesis that differential regulation of infected cell derived pro-inflammatory cytokines (IL-1beta, IL-6 and TNF-alpha) accounts for the breed-related differences in resistance and susceptibility to T. annulata infection. Other, currently unknown mechanisms may be of greater importance.     

Pollination-induced oxidative stress in floral organs of Cymbidium pendulum (Roxb.) Sw. and Cymbidium aloifolium (L.) Sw. (Orchidaceae): A biochemical investigation

A perusal of literature showed that a little is known about the metabolic changes related to senescence in orchid flowers. It was observed that unpollinated flowers of Cymbidium pendulum (Roxb.) Sw. remained fresh for 20 days and senesced within 8 days after pollination (DAP), while that of Cymbidium aloifolium (L.) Sw. took 18 days when unpollinated but showed senescence in 7 DAP. A higher level of electrolyte leakage was recorded in all the floral organs of pollinated flowers in both the species. There was a concomitant increase in levels of malondialdehyde (MDA) and hydrogen peroxide (H₂O₂); indicators of oxidative damage, in all the organs for both the species. Ascorbic acid, on the other hand, decreased significantly. Higher amount of electrolyte leakage, MDA and H₂O₂ content were recorded in C. pendulum as compared of the other species while the ascorbic acid, on the other hand, was observed to be decreased and this decrease was more in C. pendulum than C. aloifolium suggesting a higher oxidative damage to the floral organs in the former species than the latter. TIBA, i.e. tri-iodobenzoic acid (an auxin inhibitor; 0.25 μM) and silver nitrate (ethylene inhibitor; 0.25 μM) application to pollinated flowers partially prevented the elevation of oxidative damage and consequently senescence suggesting the involvement of these hormones in governing these changes in orchid flowers. Comparatively, AgNO₃ was more effective than TIBA in delaying senescence.     

1,2,4-Triazolyl functionalized allyl sulfide with antifungal potential for the control of Fusarium fujikuroi causing foot rot of rice

European Journal of Plant Pathology - The present study focuses on antifungal potential of 1,2,4-triazole derivatives of allyl sulfides. The antifungal potential of synthesized...     

Development and validation of a SYBR green real-time PCR for the quantification of porcine circovirus type 2 in serum, buffy coat, feces, and multiple tissues

The emergence of multiple genotypes of PCV2, as demonstrated by phylogenetic analysis of whole genome or capsid sequences, makes it necessary to have quantitative diagnostic assays that perform equally well on all strains. The objectives of this study were to develop and validate a novel real-time polymerase chain reaction (PCR) assay targeting the highly conserved rep gene (ORF1) and investigate the effects of diagnostic specimen choice on its performance. The assay was tested in naturally infected conventional pigs, experimentally infected gnotobiotic pigs, and plasmid-spiked negative serum, lung tissue, and feces and found to have a linear detection range of 2.2x10(3) to 2.2x10(10) copies of PCV2 per mL. The assay successfully detected and quantified PCV2 DNA in serum, buffy coat, feces, and multiple lymphoid (bronchial, mesenteric, and superficial inguinal lymph nodes; thymus; tonsil; ileal Peyer's patches; and spleen), and non-lymphoid (myocardium; lung; kidney; liver; and gluteal muscle) tissues from naturally infected pigs. Across all tissues and sera of naturally infected pigs, the mean PCV2 concentration was 3.0logs higher in wasting versus non-wasting pigs. PCV2 concentration measured by tissue culture and immunohistochemical staining in homogenized liver samples of experimentally infected gnotobiotic pigs were compared to the concentrations estimated by quantitative PCR. Similar trends were noted with increasing PCV2 concentration detected in subclinically infected to severely PMWS-affected pigs across all assays. Our diagnostic assay was developed with a conserved target sequence, and performed efficiently in quantification of PCV2 in a variety of tissues from naturally and experimentally infected pigs.     

Phytotoxic effects of Parthenium hysterophorus residues on three Brassica species

A study was conducted to assess the phytotoxicity of residues of Parthenium hysterophorus , an exotic invasive weed, towards the growth of three Brassica species ( Brassica campestris, Brassica oleracea and Brassica rapa ). The early growth of crops, measured in terms of seedling length and dry weight, was significantly reduced when grown in soil amended with varying amounts of Parthenium residues. A direct relationship was observed between the amount of residue incorporated in the soil and growth reduction. This adverse effect on Brassica crops indicates the presence of some growth-retardatory substances which are possibly released by the residues into the soil medium. In order to test this, aqueous extracts (1, 2, and 4%) of residues were prepared. It was observed that, in a laboratory bioassay, these extracts severely reduced the early growth of Brassica species, thereby indicating the presence of some water-soluble, inhibitory principles in Parthenium residues. A significant amount of the phenolics, the largest group of secondary metabolites usually implicated in allelopathy, was estimated in residue extracts, as well as in residue-incorporated soil. The phenolic content increased with increasing residue concentration, thereby showing their direct involvement in the observed growth inhibitions. Therefore, the study establishes that Parthenium residues exert an allelopathic influence on the early growth of Brassica crops by releasing water-soluble phenolics into the soil.     

大熊猫卵巢中“连体卵”的分离及其电镜观察

用切割法和切割加酶消化法对两对大熊猫卵巢进行了处理，结果在两对卵巢中别分离得到了一对典型的“连体卵”，并对其进行了电镜观察，表明大熊猫卵巢中存在“连体卵”，并初步探讨了其与大熊猫一胎产二仔的关系。