Chemical composition, intake by sheep, and in situ disappearance in cannulated cows of bermudagrass hayed at two moisture concentrations and treated with a non-viable Lactobacillus-lactic acid preservative

Bermudagrass [Cynodon dactylon (L.) Pers.] is commonly used for grazing and haying in the southern USA, but hay curing can be challenging due to frequent rainfall events during spring and early summer. An existing stand of ‘Greenfield’ bermudagrass was divided into 12 plots using a randomized complete block design with a 2×2 factorial treatment arrangement to evaluate the influence of a non-viable Lactobacillus-lactic acid preservative and moisture concentration at baling on chemical composition, intake by sheep, and in situ disappearance in cattle. At time of mowing, half of the plots in each block were either spray-treated (T) or not treated (U) with 81 mL/t forage dry matter (DM) of the preservative solution. Hay was then baled at target moisture concentrations of either 174 g/kg DM (L) or 267 g/kg DM (H). Maximum temperature and heating degree days were greater (P<0.05) from H compared with L during the 42-d storage period. An interaction between spray and moisture treatments tended (P<0.10) to affect recovery of DM; recoveries for LT (0.992) differed (P<0.10) from HT (0.913), but LU and HU were intermediate between the spray-treated hays, and did not differ from either (P>0.10). Post-storage nutritive value was largely influenced by moisture treatments only. Intake and digestibility, and in situ DM disappearance of these same hays were determined using 16 wether lambs (43 ± 3.7 kg initial BW), or six ruminally cannulated cows (617 ± 3.5 kg initial BW), respectively. Dry matter intake by sheep was not affected by either treatment factor (P>0.05), but DM digestibility and digestible DM intake were greater (P<0.05) from U compared with T. The in situ immediately soluble DM portion was greater from (P<0.05) L compared with H, but the reverse was true for the potentially degradable DM fraction. The lag time tended (P<0.10) to be greater from H compared with L. Treating bermudagrass with a non-viable Lactobacillus acidophilus-lactic acid spray product at time of baling may not offset the negative effects on forage quality and digestibility of baling bermudagrass hay at excessive moisture concentrations.     

Factors associated with the development of cross-reactive neutralizing antibodies during human immunodeficiency virus type 1 infection

The characterization of the cross-reactive, or heterologous, neutralizing antibody responses developed during human immunodeficiency virus type 1 (HIV-1) infection and the identification of factors associated with their generation are relevant to the development of an HIV vaccine. We report that in healthy HIV-positive, antiretroviral-naïve subjects, the breadth of plasma heterologous neutralizing antibody responses correlates with the time since infection, plasma viremia levels, and the binding avidity of anti-Env antibodies. Anti-CD4-binding site antibodies are responsible for the exceptionally broad cross-neutralizing antibody responses recorded only in rare plasma samples. However, in most cases examined, antibodies to the variable regions and to the CD4-binding site of Env modestly contributed in defining the overall breadth of these responses. Plasmas with broad cross-neutralizing antibody responses were identified that targeted the gp120 subunit, but their precise epitopes mapped outside the variable regions and the CD4-binding site. Finally, although several plasmas were identified with cross-neutralizing antibody responses that were not directed against gp120, only one plasma with a moderate breadth of heterologous neutralizing antibody responses contained cross-reactive neutralizing antibodies against the 4E10 epitope, which is within the gp41 transmembrane subunit. Overall, our study indicates that more than one pathway leads to the development of broad cross-reactive neutralizing antibodies during HIV infection and that the virus continuously escapes their action.     

Bacterial ecology of an old-growth douglas fir canopy

Microbial populations associated with the major substrates of the canopy of a single 70 m old-growth Douglas fir were studied to determine potential activities. Seasonal samples from bark, foliage, epiphytic moss, lichens, and litter accumulations were collected to: (a) obtain population data, (b) isolate the major groups of microorganisms present, (c) measure enzymatic activities associated with cellulose and xylan degradation, and (d) examine the potential for nitrogen fixation. We tested 562 bacterial isolates for utilization of 25 compounds associated with the canopy substrates, and for activities in nitrogen and sulfur cycle transformations. Total bacterial populations, reflecting seasonal temperature and moisture conditions, were lowest on bark and foliage [21–266×10³ colony-forming units (CFU/g)] and highest on moss and lodged litter (19–610×10⁵ CFU/g). Lichens contained intermediate numbers of bacteria (3.3–270×10⁵ CFU/g). The majority of the bacteria were classified as species ofArthrobacter, Bacillus, Flavobacterium, andXanthomonas. Isolates ofAlcaligenes (Achromobacter), Aeromonas, Chromobacterium, Micrococcus, andPseudomonas were less common. No measurable rates of nitrogen fixation attributable to free-living bacteria were detected by acetylene reduction. Eleven species in six genera of lichens containing a blue-green algal phycobiont showed positive acetylene reduction. One species,Lobaria oregana, accounted for 51% of the total lichen biomass of the canopy. Cellulase and xylanase activity was routinely detected in moss and litter samples, and less frequently in lichens. There was a strong correlation between the two activities for moss (r=0.94) and litter (r=0.81).     

Temperature-Induced Protein Conformational Changes in Barley Root Plasma Membrane-Enriched Microsomes: II. Intrinsic Protein Fluorescence

The membrane-bound proteins of barley (Hordeum vulgare L. cv Conquest) root plasma membrane-enriched microsomes displayed fluorescence typical of protein-associated trytophan residues. The protein fluorescence intensity was sensitive to variations in sample temperature. The temperature-induced decline in protein fluorescence intensity was nonlinear with slope discontinuities at about 12 and 32°C. Detergents at levels above their critical micelle concentration enhanced protein fluorescence. Glutaraldehyde reduced protein fluorescence. Protein fluorescence polarization increased at temperatures above 30°C. Both the rate of tryptophan photoionization and the fluorescence intensity of the photoionization products suggested alterations in membrane protein conformation between 12 and 32°C. The quenching of the intrinsic protein fluorescence by acrylamide and potassium iodide indicated changes in accessibility of the extrinsic agents to the protein tryptophan residues beginning at about 14°C. The results indicate thermally induced changes in the dynamics of the membrane proteins over the temperature range of 12 to 32°C which could account for the complex temperature dependence of the barley root plasma membrane ATPase.     

心脏特异表达LMNA^E82K转基因小鼠的建立

目的建立心脏特异表达LMNAE82K转基因小鼠,为研究LMNAE82K与心肌病发病机制的关系提供工具动物。方法把LMNAE82K基因插入α-MHC启动子下游,构建转基因表达载体,显微注射法建立C57BL/6JLMNAE82K转基因小鼠,PCR鉴定转基因小鼠的基因型,采用Western Blot鉴定LMNAE82K在心脏组织中的表达,H＆E染色和超声检测转基因小鼠心脏的病理改变。结果建立了2个心脏组织特异表达LMNAE82K的转基因小鼠品系。超声检查显示转基因小鼠心室壁变薄,收缩期容积和舒张期容积增加,射血分数及短轴缩短率降低。结论LMNAE82K转基因小鼠具有LMNAE82K引起的家族性扩心病有类似的病理变化,为研究LMNAE82K与心肌病发病机制的关系的研究提供了有价值的疾病动物模型。     

A molecular mechanical study of netropsin-DNA interactions

We present molecular mechanical calculations on the complexes of netropsin with dA₆·dT₆, d(TATATA)₂, d(CGCGCG)₂, and d(CGCGAATTCGCG)₂. The complexes were model built using computer graphics and then completely energy refined. Our calculations are consistent with the observed AT preference for netropsin and suggest that mixed sugar pucker geometries should be more stable than uniform in netropsin complexes with poly[d(A-T)]·poly[d(A-T)] and poly(dA)·poly(dt). The netropsin·d(TATATA) and netropsin·dA₆·dT₆ complexes are significantly different in structure, leading to a possible reason why the observed thermodynamics of netropsin-association with poly[d(A-T)]·poly[d(A-T)] and with poly(dA)·poly(dT) are so different. We also model built and energy refined a structure of netropsin-d(CGCGAATTCGCG)₂ using as a guide the nmr data of Patel [(1982) Proc. Natl. Acad. Sci. USA, 79 , 6424–6428] and found a three-dimensional structure qualitatively consistent with the NOE enhancements observed by him. After our calculations were completed, we learned of an x-ray structure of a netropsin:d(CGCGAATTCGCG)₂ complex, and we compared the structure found in our calculation with the x-ray structure.