Virulent Mycobacterium bovis Beijing Strain Activates the NLRP7 Inflammasome in THP-1 Macrophages

Mycobacterium bovis is the causative agent of tuberculosis in a wide range of mammals, including humans. Macrophages are the first line of host defense. They secrete proinflammatory cytokines, such as interleukin-1 beta (IL-1β), in response to mycobacterial infection, but the underlying mechanisms by which human macrophages are activated and release IL-1β following M. bovis infection are poorly understood. Here we show that the ‘nucleotide binding and oligomerization of domain-like receptor (NLR) family pyrin domain containing 7 protein’ (NLRP7) inflammasome is involved in IL-1β secretion and caspase-1 activation induced by M. bovis infection in THP-1 macrophages. NLRP7 inflammasome activation promotes the induction of pyroptosis as well as the expression of tumor necrosis factor alpha (TNF-α), Chemokine (C-C motif) ligand 3 (CCL3) and IL-1β mRNAs. Thus, the NLRP7 inflammasome contributes to IL-1β secretion and induction of pyroptosis in response to M. bovis infection in THP-1 macrophages.     

Double-inducible gene activation system for caspase 3 and 9 in epidermis

Expression of genes with tight and precise temporal and spatial control is desired in a wide variety of applications ranging from cultured cells and transgenic animals to gene therapy. While current inducible systems, such as RU486 and chemical inducers of dimerization (CID), have improved earlier inducible models (Gossen et al., 1995, Science. 268:1766-1769; Wang et al., 1994, Proc Natl Acad Sci USA 91:8180-8184), no single system is perfect at present. One potential drawback of these systems is leakage of transgene expression, causing limitations of each system. We have developed an inducible model containing both RU486 and CID systems, which in addition to inducing caspase activation, has potential applicability specifically to other genes encoding proteins that require a dimerization event for activation. This Double-Inducible Gene Activation System generates two barriers for the target gene expression and protein activation thereby minimizing leakage.     

Enzymatic Dehalogenation of Pentachlorophenol by <Emphasis Type="Italic">Pseudomonas fluorescens</Emphasis> of the Microbial Community from Tannery Effluent

Four different bacterial isolates obtained from a stable bacterial consortium were capable of utilizing pentachlorophenol (PCP) as sole carbon and energy source. The consortium was developed by continuous enrichment in the chemostat. The degradation of PCP by bacterial strain was preceded through an oxidative route as indicated by accumulation of tetrachloro-ρ-hydroquinone and dichlorohydroquinone as determined by high performance liquid chromatography (HPLC). Among the four isolates, Pseudomonas fluorescens exhibited maximum degradation capability and enzyme production. PCP-monooxygenase enzyme was extracted from culture extract and fractionated by DEAE-cellulose ion exchange chromatography. The molecular weight of the enzyme, purified from Pseudomonas fluorescens, determined by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and gel filtration chromatography was found to be 24,000 Da. Received: 22 July 2002 / Accepted: 23 September 2002     

Variation in the ribosomal internal transcribed spacers and 5.8S rDNA among five species of Acropora (Cnidaria; Scleractinia): patterns of variation consistent with reticulate evolution

The ITS sequences of Acropora spp. are the shortest so far identified in any metazoan and are among the shortest seen in eukaryotes; ITS1 was 70-80 bases, and ITS2 was 100-112 bases. The ITS sequences were also highly variable, but base composition and secondary structure prediction indicate that divergent sequence variants are unlikely to be pseudogenes. The pattern of variation was unusual in several other respects: (1) two distinct ITS2 types were detected in both A. hyacinthus and A. cytherea, species known to hybridize in vitro with high success rates, and a putative intermediate ITS2 form was also detected in A. cytherea; (2) A. valida was found to contain highly (29%) diverged ITS1 variants; and (3) A. longicyathus contained two distinct 5.8S rDNA types. These data are consistent with a reticulate evolutionary history for the genus Acropora.      

Cardiac Specific Expression of Threonine 5 to Alanine Mutant Sarcolipin Results in Structural Remodeling and Diastolic Dysfunction

The functional importance of threonine 5 (T5) in modulating the activity of sarcolipin (SLN), a key regulator of sarco/endoplasmic reticulum (SR) Ca²⁺ ATPase (SERCA) pump was studied using a transgenic mouse model with cardiac specific expression of threonine 5 to alanine mutant SLN (SLNT5A). In these transgenic mice, the SLNT5A protein replaces the endogenous SLN in atria, while maintaining the total SLN content. The cardiac specific expression of SLNT5A results in severe cardiac structural remodeling accompanied by bi-atrial enlargement. Biochemical analyses reveal a selective downregulation of SR Ca²⁺ handling proteins and a reduced SR Ca²⁺ uptake both in atria and in the ventricles. Optical mapping analysis shows slower action potential propagation in the transgenic mice atria. Doppler echocardiography and hemodynamic measurements demonstrate a reduced atrial contractility and an impaired diastolic function. Together, these findings suggest that threonine 5 plays an important role in modulating SLN function in the heart. Furthermore, our studies suggest that alteration in SLN function can cause abnormal Ca²⁺ handling and subsequent cardiac remodeling and dysfunction.     

Global synthesis of the temperature sensitivity of leaf litter breakdown in streams and rivers

Streams and rivers are important conduits of terrestrially derived carbon (C) to atmospheric and marine reservoirs. Leaf litter breakdown rates are expected to increase as water temperatures rise in response to climate change. The magnitude of increase in breakdown rates is uncertain, given differences in litter quality and microbial and detritivore community responses to temperature, factors that can influence the apparent temperature sensitivity of breakdown and the relative proportion of C lost to the atmosphere vs. stored or transported downstream. Here, we synthesized 1025 records of litter breakdown in streams and rivers to quantify its temperature sensitivity, as measured by the activation energy (E_a, in eV). Temperature sensitivity of litter breakdown varied among twelve plant genera for which E_a could be calculated. Higher values of E_a were correlated with lower‐quality litter, but these correlations were influenced by a single, N‐fixing genus (Alnus). E_a values converged when genera were classified into three breakdown rate categories, potentially due to continual water availability in streams and rivers modulating the influence of leaf chemistry on breakdown. Across all data representing 85 plant genera, the E_a was 0.34 ± 0.04 eV, or approximately half the value (0.65 eV) predicted by metabolic theory. Our results indicate that average breakdown rates may increase by 5–21% with a 1–4 °C rise in water temperature, rather than a 10–45% increase expected, according to metabolic theory. Differential warming of tropical and temperate biomes could result in a similar proportional increase in breakdown rates, despite variation in E_a values for these regions (0.75 ± 0.13 eV and 0.27 ± 0.05 eV, respectively). The relative proportions of gaseous C loss and organic matter transport downstream should not change with rising temperature given that E_a values for breakdown mediated by microbes alone and microbes plus detritivores were similar at the global scale.     

The Enhancement of Soil Fertility,Dry Matter Transport and Accumulation,Nitrogen Uptake and Yield in Rice via Green Manuring

Readily available chemical fertilizers have resulted in a decline in the use of organic manure (e.g., green manures), a traditionally sustainable source of nutrients. Based on this, we applied urea at the rate of 270 kg ha⁻¹ with and without green manure in order to assess nitrogen (N) productivity in a double rice cropping system in 2017. In particular, treatment combinations were as follows: winter fallow rice-rice (WF-R-R), milk vetch rice-rice (MV-R-R), oil-seed rape rice-rice (R-R-R) and potato crop rice-rice (P-R-R). Results revealed that green manure significantly (p ≤ 0.05) improved the soil chemical properties and net soil organic carbon content increased by an average 117.47%, total nitrogen (N) by 28.41%, available N by 26.64%, total phosphorus (P) by 37.77%, available P by 20.48% and available potassium (K) by 33.10% than WF-R-R, however pH was reduced by 3.30% across the seasons. Similarly, net dry matter accumulation rate enhanced in green manure applied treatments and ranked in order: P-R-R > R-R-R > MV-R-R > WF-R-R. Furthermore, the total leaf dry matter transport (t ha⁻¹ ) for the P-R-R in both seasons was significantly higher by an average 11.2%, 7.2% and 36 % than MV-R-R, R-R-R, and WF-R-R, respectively. In addition, net total nitrogen accumulation (kg ha⁻¹ ) was found higher in green manure applied plots compared to the control. Yield and yield attributed traits were observed maximum in green manure applied plots, with treatments ranking as follows: P-R-R > R-R-R > MV-R-R > WF-R-R. Thus, results obtained highlight ability of green manure to sustainably improve soil quality and rice yield.     

Hepatic protein kinase C: translocation stimulated by prolactin and partial hepatectomy

Prolactin stimulates a hepatotrophic response similar to that caused by phorbol esters or partial hepatectomy in rats. Since phorbol esters, which activate protein kinase C, mimic prolactin action in liver, the relationship between prolactin administration and subsequent hepatic protein kinase C translocation was assessed. Prolactin administration rapidly stimulated a 4-fold elevation of membrane protein kinase C activity. The effect of prolactin on hepatic protein kinase C was specific for lactogenic hormones but could be duplicated by phorbol esters. Further, an increase in serum prolactin was demonstrated subsequent to partial hepatectomy and preceding hepatic protein kinase C translocation. Therefore, translocation of hepatic protein kinase C appears important for hepatic proliferation in response to prolactin administration and to partial hepatectomy.