Release of Rhizobium spp. from Tropical Soils and Recovery for Immunofluorescence Enumeration

Limitations associated with immunofluorescence enumeration of bacteria in soil derive largely from the efficiency with which cells can be separated from soil particles and collected on membrane filters for staining. Many tropical soils fix added bacteria tightly, resulting in low recoveries. Eight soils, representative of three of the major soil orders found in the tropics (oxisols, vertisols, and inceptisols), were tested for recovery of added Rhizobium strains. All except one Hawaiian andept (Typic Eutrandept) yielded recoveries ranging from <1 to 13%. Recovery from the andept was 100%. In soil-sand mixtures, addition of only a small amount of soil caused a dramatic decrease in recovery of added rhizobia. Increasing the soil content of the mixture from 0% (10 g of sand) to 50% (5 g of soil-5 g of sand) reduced recoveries from >90 to <1%. Varying the ionic strength and pH of the extracting solution did not cause marked increases in recovery. Protein solutions, ethylenediaminetetraacetate, and NaHCO₃, on the other hand, improved release of bacteria. We report a modification to the usual membrane filter immunofluorescence procedure which yielded consistently high and reproducible recovery (coefficient of variation, 30%) of rhizobia from several tropical soils. In the modified procedure, partially hydrolyzed gelatin, diluted in ammonium phosphate, was used to suspend the soil. This caused dispersion of the soil and release of the bacteria from soil flocs. The efficiency of recovery of Rhizobium spp. from several tropical and two temperate soils remained high as the content of these soils in soil-sand mixtures was increased from 0 to 100%. The modified membrane filter immunofluorescence procedure was used to follow the growth of a strain of chickpea (Cicer arietinum) Rhizobium in a sterilized oxisol. The results showed a close agreement with viable counts at different stages during the growth cycle. Diluent for the hydrolyzed gelatin also had a marked effect on recovery. The efficiency of release of Rhizobium spp. from an oxisol was in the following order for the diluents used: 0.1 M (NH₄)₂HPO₄ > 0.1 M Na₂HPO₄ = 0.1 M sodium-phosphate-buffered saline (pH 7.2) > 0.2 M NH₄Cl > 0.2 KCl > NaCl = LiCl > water.     

Methionine degradation inCandida utilis

Incubation of cell-free extracts ofC. utilis and its ethionine-resistant mutants with methionine, aspartate and alanine, and with various acceptor oxo-acids showed the presence of several transamination activities (Glu-Asp, Ala-Glu, Met-Glu, Met-Ala, Ala-Asp, but not Met-Asp). The lack of utilization of methionine by mec mutants appears to be due to a transport lesion.     

Sucrose:sucrose fructosyltransferase and fructan:fructan fructosyltransferase from allium cepa

Sucrose: sucrose fructosyltransferase and fructan:fructan fructosyltransferase were isolated from the inner leaf bases of bulbing onion plants (Allium cepa) and separated by gel filtration on Bio-Gel P-150. Sucrose:sucrose fructosyltransferase produced only one trisaccharide, 1^F-fructosylsucrose, from sucrose. Fructan:fructan fructosyltransferase produced tetrasaccharide and higher polymers from trisaccharide. The trisaccharide found in the greatest concentration in onion, 6^G-fructosylsucrose, was produced from 1^F-fructosylsucrose by fructan:fructan fructosyltransferase and was not a product of sucrose:sucrose fructosyltransferase.     

On the effect of glucono-δ-lactone on the yeast Pichia polymorpha

Investigation has been made into the action of glucono--lactone on living cells of Pichia polymorpha in relation to the uptake of D-(U-¹⁴C) glucose, and the incorporation of (2-¹⁴C) uracil and L-(U-¹⁴C)-threonine into RNA and protein respectively. Other factors such as the action of glucono--lactone on cell morphology and on enzymic synthesis have also been studied. The action of this compound on -glucanase has been found to take place in the hydrolytic power and not in the synthesis.     

Synthesis of β-glucanase and other extracellular proteins by cells and protoplasts of the yeast Pichia polymorpha

The synthesis of β-glucanase either by cells or by protoplasts of the yeast Pichia polymorpha has been found to occur in the presence of 2-deoxy-d-glucose in the growth medium. On the other hand, the synthesis of typical extracellular proteins such as invertase and acid phosphatase is strongly affected by the presence of the drug. The degree of inhibition is, however, directly related to the 2-deoxy-d-glucose concentration.     

Laying low: Rugged lowland rainforest preferred by feral cats in the Australian Wet Tropics

Invasive mesopredators are responsible for the decline of many species of native mammals worldwide. Feral cats have been causally linked to multiple extinctions of Australian mammals since European colonization. While feral cats are found throughout Australia, most research has been undertaken in arid habitats, thus there is a limited understanding of feral cat distribution, abundance, and ecology in Australian tropical rainforests. We carried out camera‐trapping surveys at 108 locations across seven study sites, spanning 200 km in the Australian Wet Tropics. Single‐species occupancy analysis was implemented to investigate how environmental factors influence feral cat distribution. Feral cats were detected at a rate of 5.09 photographs/100 days, 11 times higher than previously recorded in the Australian Wet Tropics. The main environmental factors influencing feral cat occupancy were a positive association with terrain ruggedness, a negative association with elevation, and a higher affinity for rainforest than eucalypt forest. These findings were consistent with other studies on feral cat ecology but differed from similar surveys in Australia. Increasingly harsh and consistently wet weather conditions at higher elevations, and improved shelter in topographically complex habitats may drive cat preference for lowland rainforest. Feral cats were positively associated with roads, supporting the theory that roads facilitate access and colonization of feral cats within more remote parts of the rainforest. Higher elevation rainforests with no roads could act as refugia for native prey species within the critical weight range. Regular monitoring of existing roads should be implemented to monitor feral cats, and new linear infrastructure should be limited to prevent encroachment into these areas. This is pertinent as climate change modeling suggests that habitats at higher elevations will become similar to lower elevations, potentially making the environment more suitable for feral cat populations.     

Division of labor within the DNA damage tolerance system reveals non-epistatic and clinically actionable targets for precision cancer medicine

Crosslink repair depends on the Fanconi anemia pathway and translesion synthesis polymerases that replicate over unhooked crosslinks. Translesion synthesis is regulated via ubiquitination of PCNA, and independently via translesion synthesis polymerase REV1. The division of labor between PCNA-ubiquitination and REV1 in interstrand crosslink repair is unclear. Inhibition of either of these pathways has been proposed as a strategy to increase cytotoxicity of platinating agents in cancer treatment. Here, we defined the importance of PCNA-ubiquitination and REV1 for DNA in mammalian ICL repair. In mice, loss of PCNA-ubiquitination, but not REV1, resulted in germ cell defects and hypersensitivity to cisplatin. Loss of PCNA-ubiquitination, but not REV1 sensitized mammalian cancer cell lines to cisplatin. We identify polymerase Kappa as essential in tolerating DNA damage-induced lesions, in particular cisplatin lesions. Polk-deficient tumors were controlled by cisplatin treatment and it significantly delayed tumor outgrowth and increased overall survival of tumor bearing mice. Our results indicate that PCNA-ubiquitination and REV1 play distinct roles in DNA damage tolerance. Moreover, our results highlight POLK as a critical TLS polymerase in tolerating multiple genotoxic lesions, including cisplatin lesions. The relative frequent loss of Polk in cancers indicates an exploitable vulnerability for precision cancer medicine.     

四川省大熊猫保护地生态安全及其时空演变

大熊猫是生态保护的旗舰物种,保护大熊猫及其栖息地具有重要的示范作用和科学价值。中国致力于建设以国家公园为主体的自然保护地体系,分析大熊猫保护地生态安全,为建立大熊猫国家公园提供理论依据,具有重要意义。以四川省为例,选取被划入或将来可能被划入大熊猫保护地的典型县域,构建大熊猫保护地生态安全评价指标体系,采用熵权法、层次分析法进行综合评价,并结合空间相关性分析,从时间和空间两个维度对大熊猫保护地生态安全状况进行时空动态分析。结果表明：（1）2003-2017年四川省大熊猫保护地生态安全综合指数呈波动性上升趋势,生态环境改善显著,但仍有部分县域出现不同程度的环境恶化现象。（2）各县域生态安全状况表现出较强的差异性,并且相邻县域生态安全指数存在显著的空间正相关性和空间集聚性。（3）大熊猫保护地生态安全与周边社会经济发展之间矛盾依然尖锐,相邻县域之间的经济发展和生态保护状况相互影响的作用明显,需联合治理,共同促进生态平衡,提高大熊猫及其栖息地保护效果。     

用猴艾滋病D型逆转录病毒筛选抗艾滋病药物实验模型的建立及应用

以药物对合胞体形成的抑制和对细胞的毒性作为检测指标,应用猴艾滋病D型逆转录病毒（SRV）／Raji细胞系统建立了体外筛选抗艾滋病药物的实验模型。AZT和天花粉蛋白可显著地抑制SRV诱导的合胞体形成,它们的选择指数（SI）分别为数13500和8812。用该模型筛选了46种来源于天然资源的化合物,结果表明有11种天然化合物有明显的抗病毒活性。     

Using electroporation and lipid-mediated transfection of GFP-expressing plasmids to label embryonic avian cells for vital confocal and two-photon microscopy

Fluorescent proteins have emerged as an ideal fluorescent marker for studying cell morphologies in vital systems. These proteins were first applied in whole organisms with established germ-line transformation protocols, but now it is possible to label cells with fluorescent proteins in other organisms. Here we present two ways to introduce GFP expressing plasmids into avian embryos for vital confocal and two-photon imaging. First, electroporation is a powerful approach to introduce GFP into the developing neural tube, offering several advantages over dye labeling. Second, we introduce a new lipid-based transfection system for introducing plasmid DNA directly to a small group of injected cells within live, whole embryos. These complementary approaches make it possible to transfect a wide-range of cell types in the avian embryo and the bright, stable, uniform expression of GFP offers great advantages for vital fluorescence imaging.