Mitigation of N2O emission from granular organic fertilizer with alkali- and salt-resistant plant growth-promoting rhizobacteria.

AIM: Organic fertilizer application significantly stimulates nitrous oxide (N2O) emissions from agricultural soils. Plant growth-promoting rhizobacteria (PGPR) strains are the core of bio-fertilizer or bio-organic fertilizer, while their beneficial effects are inhibited by environmental conditions, such as alkali and salt stress observed in organic manure or soil. This study aims to screen alkali- and salt-resistant PGPR that could mitigate N2O emission after applying strain-inoculated organic fertilizer. METHODS AND RESULTS: Among the 29 candidate strains, 11 (7 Bacillus spp., 2 Achromobacter spp., 1 Paenibacillus sp., and 1 Pseudomonas sp.) significantly mitigated N2O emissions from the organic fertilizer after inoculation. Seven strains were alkali tolerant (pH 10) and five were salt tolerant (4% salinity) in pure culture. Seven strains were selected for further evaluation in two agricultural soils. Five of these seven strains could significantly decrease the cumulative N2O emissions from Anthrosol, while six could significantly decrease the cumulative N2O emissions from Cambisol after the inoculation into the granular organic fertilizer compared with the non-inoculated control. CONCLUSIONS: Inoculating alkali- and salt-resistant PGPR into organic fertilizer can reduce N2O emissions from soils under microcosm conditions. Further studies are needed to investigate whether these strains will work under field conditions, under higher salinity, or at different soil pH.

Genome Sequence of Arthrobacter sp. UKPF54-2, a Plant Growth-Promoting Rhizobacterial Strain Isolated from Paddy Soil.

Arthrobacter sp. strain UKPF54-2, a plant growth-promoting rhizobacterium having the potential ability to control fungal and bacterial pathogens, was isolated from paddy soil in Kumamoto, Japan. We report here the whole-genome sequence of this strain.

Developmental exposure to mercury chloride impairs social behavior in male offspring dependent on genetic background and maternal autoimmune environment.

To date, the connection between inorganic mercury (Hg) and social behavior remains incompletely understood. The aim of this study was to investigate the influence of maternal autoimmunity by inorganic Hg (Hg2+) exposure on social behavior of offspring. Wild-type (WT) and immunoglobulin deficient (Ig-/-) B10.S dams fertilized by male WT B10.S or SJL mice were treated with 50 µM Hg chloride (HgCl2). Non-pregnant female WT B10.S mice were used to investigate factors regulating HgCl2-induced autoimmunity to brain. HgCl2 selectively impaired social behavior in male offspring, but not female offspring from WT B10.S dams × male SJL, in that only male offspring displayed reduced time distribution with the stranger mouse, decreased sniffing to the stranger mouse and increased self-grooming. HgCl2 did not disrupt social behavior of male or female offspring from WT B10.S dams × male WT B10.S or Ig-/- B10.S dams × male SJL. The offspring from WT and Ig-/- B10.S dams × male SJL had equivalent autoimmunity to brain antigens during HgCl2 exposure, indicating that maternal, but not offspring-derived anti-brain antibodies (Ab) impaired social behavior of the offspring. Non-pregnant WT B10.S mice treated with HgCl2 had increased anti-brain Ab dependent on increase in CD4 T cell activation and IFNγ signaling to macrophages. IFNγ interaction with macrophages drove B cells and plasma cells to produce IgG. Therefore, HgCl2 selectively impaired social behavior in males with certain genetic background via maternally derived anti-brain Ab production, thus providing a novel insight into our current understanding of Hg toxicity.

Mercury impact on hematopoietic stem cells is regulated by IFNγ-dependent bone marrow-resident macrophages in mice.

Although immunotoxic effects of mercury (Hg) have been extensively investigated, the influence of Hg on hematopoietic stem cells (HSC) remains elusive. The aim of this study was to investigate the effects of Hg on HSC. B10.S (H-2s) and DBA/2 mice (H-2d) were treated with Hg chloride (25, 50 or 100 µM HgCl2) or methyl Hg (1.25, 3.75 or 6.25 µM MeHg) via drinking water for 4 weeks, and thereafter, HSC in the bone marrow (BM) were evaluated. The number of HSC in B10.S mice was increased after treatment with 50 µM HgCl2 and decreased after treatment with 100 µM HgCl2; the number of HSC in DBA/2 mice was reduced after treatment with 50 µM HgCl2 and unaffected after treatment with 25 µM HgCl2. These effects from the HgCl2 treatments were associated with alterations of HSC proliferation, IFNγ expression and BM-resident macrophages. In vivo neutralization of IFNγ diminished the HgCl2-driven HSC proliferation, and in vivo replenishment of recombinant IFNγ eliminated the HgCl2 suppression of HSC proliferation and allowed HgCl2 enhancement of proliferation, suggesting a pivotal role of IFNγ in HSC proliferation regulated by HgCl2. In vivo depletion of macrophages and an in vitro co-culture assay indicated that BM-resident macrophages promoted HSC proliferation during HgCl2 exposure. Furthermore, the induction of BM-resident macrophages was critically dependent on IFNγ. In contrast, MeHg did not influence HSC in B10.S or DBA/2 mice. Collectively, HgCl2, but not MeHg, affects HSC through regulating IFNγ-dependent BM-resident macrophages in mice. These findings reveal a previously unknown toxicity of Hg.

Excess pneumonia and influenza mortality attributable to seasonal influenza in subtropical Shanghai, China.

BACKGROUND: Disease burden attributable to influenza is substantial in subtropical regions. Our study aims to estimate excess pneumonia and influenza (P&I) mortality associated with influenza by subtypes/lineages in Shanghai, China, 2010-2015. METHODS: Quasi-Poisson regression models were fitted to weekly numbers of deaths from causes coded as P&I for Shanghai general and registered population. Three proxies for influenza activity were respectively used as an explanatory variable. Long-term trend, seasonal trend and absolute humidity were adjusted for as confounding factors. The outcome measurements of excess P&I mortality associated with influenza subtypes/lineages were derived by subtracting the baseline mortality from fitted mortality. RESULTS: Excess P&I mortality associated with influenza were 0.22, 0.30, and 0.23 per 100,000 population for three different proxies in Shanghai general population, lower than those in registered population (0.34, 0.48, and 0.36 per 100,000 population). Influenza B (Victoria) lineage did not contribute to excess P&I mortality (P = 0.206) while influenza B (Yamagata) lineage did (P = 0.044). Influenza-associated P&I mortality was high in the elderly population. CONCLUSIONS: Seasonal influenza A virus had a higher P&I mortality than influenza B virus, while B (Yamagata) lineage is the dominant lineage attributable to P&I mortality.

Lead Transiently Promotes Granulocyte-Macrophage Progenitor Differentiation and Subsequently Suppresses Common Myeloid Progenitor Differentiation.

Lead (Pb) is a toxic heavy metal affecting human health; it is known to be harmful to various organs or systems, yet the mechanisms by which Pb influences immune cell development remain to be defined. In this study, we show that Pb exposure (1250 ppm via drinking water) selectively impacted the development of myeloid cells (myelopoiesis). After Pb treatment of adult C57BL/6 mice, the numbers of granulocyte-macrophage progenitors (GMP) were consistently reduced, whereas the numbers of myeloid cells were increased at week (wk) 1 and decreased at wk8 after initiating the Pb exposure. Functional assays indicate that Pb accelerated GMP differentiation in a reactive oxygen species-dependent manner after treatment for 1 week and inhibited common myeloid progenitor differentiation by upregulating interferon regulatory factor 8 (IRF8) expression after treatment for 8 weeks. Consistent with the distinct Pb influences on myeloid cells observed at wk1 and wk8, Pb caused an inflammatory environment in vivo at wk8, but not at wk1. Furthermore, like the observations in mice during the Pb exposure, bloods from humans occupationally exposed to Pb had their numbers of monocytes, neutrophils and GMP negatively associated with the Pb concentration, whereas IRF8 expression in common myeloid progenitor, but not GMP, was positively correlated with the Pb concentration. These data suggest an occupationally relevant level of Pb exposure preferentially influences myelopoiesis involving reactive oxygen species and IRF8, which may contribute to the current understanding of the hematopoietic toxicology of Pb.

Cadmium modulates hematopoietic stem and progenitor cells and skews toward myelopoiesis in mice.

The heavy metal cadmium (Cd) is known to modulate immunity and cause osteoporosis. However, how Cd influences on hematopoiesis remain largely unknown. Herein, we show that wild-type C57BL/6 (B6) mice exposed to Cd for 3months had expanded bone marrow (BM) populations of long-term hematopoietic stem cells (LT-HSCs), common myeloid progenitors (CMPs) and granulocyte-macrophage progenitors (GMPs), while having reduced populations of multipotent progenitors (MPPs) and common lymphoid progenitors (CLPs). A competitive mixed BM transplantation assay indicates that BM from Cd-treated mice had impaired LT-HSC ability to differentiate into mature cells. In accordance with increased myeloid progenitors and decreased lymphoid progenitors, the BM and spleens of Cd-treated mice had more monocytes and/or neutrophils and fewer B cells and T cells. Cd impaired the ability of the non-hematopoietic system to support LT-HSCs, in that lethally irradiated Cd-treated recipients transplanted with normal BM cells had reduced LT-HSCs after the hematopoietic system was fully reconstituted. This is consistent with reduced osteoblasts, a known critical component for HSC niche, observed in Cd-treated mice. Conversely, lethally irradiated control recipients transplanted with BM cells from Cd-treated mice had normal LT-HSC reconstitution. Furthermore, both control mice and Cd-treated mice that received Alendronate, a clinical drug used for treating osteoporosis, had BM increases of LT-HSCs. Thus, the results suggest Cd increase of LT-HSCs is due to effects on HSCs and not on osteoblasts, although, Cd causes osteoblast reduction and impaired niche function for maintaining HSCs. Furthermore, Cd skews HSCs toward myelopoiesis.

Microwave heating of long-term formalin-fixed surgical pathology specimens improves quality of extracted DNA.

A large amount of DNA of high quality is essential for molecular analysis. The amount of DNA in routine paraffin sections is small. Surgical specimens retained in formalin for the long-term (several months) left over from the sampling required for wax embedding can be referred to as "long-term formalin-fixed" specimens, and clearly this material is a rich source of DNA; however, it is difficult to extract. In the current study, we designed a microwave-heating method for DNA extraction from these specimens. We found that the heating procedure achieved greater DNA yields than a common nonheating method used for comparison (DNA contents mean±SD, heating 2.16±0.95 µg/µL vs. common 1.75±0.90 µg/µL, P<0.05). Fluorescence multiplex polymerase chain reaction-capillary electrophoresis successfully detected microsatellite instability (MSI) in the DNA samples derived from this heating procedure (98.4%, 689 of 700 sample tests), at significantly higher levels than from the conventional method (82.3%, 247 of 300 sample tests, P<0.05). We identified 10 (14.3%) MSI with high frequency and 6 (8.6%) MSI with low frequency colorectal cancers. MSI with high frequency cancers showed distinct clinicopathologic features including higher incidence of right-sided location, high histologic grade, mucin-production, and prominent intraepithelial lymphocyte infiltration. We concluded that the microwave-heating method was efficient for DNA isolation from long-term formalin-fixed tissue samples. The successful fluorescence multiplex polymerase chain reaction-capillary electrophoresis analysis in these samples might facilitate MSI detection in clinical practice.

The association between metabolic abnormality and endometrial cancer: a large case-control study in China.

OBJECTIVES: To investigate the association between insulin-resistance-related conditions (comprising the metabolic syndrome) and endometrial cancer risk in Chinese women. METHODS: We conducted a large case-control study including 942 endometrial cancers and 1721 controls on a Chinese population. The relative endometrial cancer risks from various factors were calculated by the x(2) test. Menopausal status and BMI were applied as potential confounders to analyze the joint effects with other factors. RESULTS: We found that overweight/obesity, hypertension, diabetes mellitus and glucose metabolic disturbance were all associated with endometrial cancer risk. Effective medication on diabetes can significantly decrease cancer risk (uncontrolled diabetes: RR=5.563, 95% CI=2.406-12.859, p<0.001; controlled diabetes: RR=1.331, 95% CI=0.529-3.352, p>0.05) as compared with normal controls. Serum lipids were also found to be linked to endometrial cancer risk: positive correlations were present with total serum cholesterol, triglycerides, low-density lipoprotein cholesterol and dyslipidaemia, while a negative correlation was found with high-density lipoprotein cholesterol. We also observed an elevated risk for type I endometrial cancer (OR=1.839, 95% CI=1.539-2.197, p<0.001) in women with BMI>or=24.58 versus those with BMI<24.58, but not for type II cancer (OR=1.092, 95% CI=0.969-1.231, p>0.05). CONCLUSIONS: Our findings support the hypothesis that metabolic abnormalities or insulin resistance may promote the initiation and progression of endometrial cancer. The effective treatment of type 2 diabetes might contribute to endometrial cancer prevention.

Cloning of a novel ovalbumin gene from quail oviduct and its heterologous expression in Pichia pastoris.

An ovalbumin gene was cloned from Chinese quail (Coturnix coturnix) oviduct by RT-PCR and then inserted into the P. pastoris genome under the control of the methanol inducible 5' alcohol oxidase (AOX) promoter. The recombinant P. pastoris strain was demonstrated to be able to efficiently secrete quail ovalbumin by ELISA analysis using a polyclonal antibody raised against quail ovalbumin. The results showed that induction by 0.75% methanol for 48 h led to the synthesis of secreted quail ovalbumin up to a yield of 5.45 g l(-1). The recombinant ovalbumin was further purified into homogeneity through ion exchange and gel filtration chromatography and SDS-PAGE analysis revealed that, compared to natural ovalbumin, the recombinant ovalbumin could have been glycosylated to the similar extent by P. pastoris.

[Platelet count of peripheral blood is associated with invasion and metastasis of esophageal carcinoma].

OBJECTIVE: To analyze the association between the platelet count of peripheral blood and clincopathologic parameters of esophageal carcinoma. METHODS: Platelets of peripheral blood were measured in 415 cases of esophageal carcinoma and 325 healthy subjects as control. The correlation of platelet counts and clinicopathological features of cancer was analyzed. RESULT: Platelet count in patients with esophageal carcinomas (286+/-88)x10(9)/L was significantly higher than that in the control subjects [(204+/-114)x10(9)/L, P<0.05 ]. Increased platelet counts (>300 x 10(9)/L) was significantly associated with tumor infiltration and lymph node metastasis in patients with esophageal cancer (P<0.05). CONCLUSION: Platelet count of peripheral blood might be associated with the development and progression of esophageal cancer.

A method for filling in the cohesive ends of double-stranded DNA using Pfu DNA polymerase.

The present paper reports a highly efficient method of making blunt ends from cohesive ends of double-stranded DNA. Klenow fragment and Pfu DNA polymerases were used to fill in the cohesive ends. Since the transformation efficiency can directly reflect the filling-in efficiency, similar ligation and transformation conditions were used, and the filling-in efficiency was compared with the corresponding transformation efficiency. The results indicate that the filling-in efficiency of Pfu DNA polymerase was 1.96 times that of Klenow fragment and its efficiency was markedly higher than that of Klenow fragment (P<0.01). The optimization experiments on reaction conditions indicate, when the pH is 8.5 and the temperature is 74 degrees C, that the filling-in efficiency was highest upon using a buffer containing 3 mM MgSO4 and 300 microM dNTP.