Intermediate-Scale, Semicontinuous Solid-Phase Fermentation Process for Production of Fuel Ethanol from Sweet Sorghum

A novel, semicontinuous solid-phase fermentation system was used to produce fuel ethanol from sweet sorghum. The process was at an intermediate scale. In the process, dried and shredded sweet sorghum was rehydrated to 70% moisture, acidified to pH 2.0 to 3.0, and either pasteurized (12 h at 70 to 80°C) or not pasteurized before spray inoculation with a broth culture of Saccharomyces cerevisiae. Fermented pulp exited the semicontinuous fermentor after a retention time of 72 h and contained approximately 6% (vol/vol) ethanol. Ethanol yields from dry sweet sorghum were 176 to 179 liters/10³ kg (85% of theoretical). Production costs for a greatly scaled-up (×1,400) conceptual version of this system were projected by calculation to average $0.47/liter for 95% ethanol. The calculated energy balance (energy output/energy input ratio) was estimated to be 1.05 when pasteurization was included and 1.31 when pasteurization was omitted. In calculating the energy balances, the output energy of the protein feed byproduct and the input energy for growing the sweet sorghum were not considered. A design for the scaled-up plant (farm scale) is provided.     

Purification and Properties of In Vitro–produced Anthrax Toxin Components

The three components of the toxin of Bacillus anthracis, edema factor (EF), protective antigen (PA), and lethal factor (LF), were purified 197-, 156-, and 1,025- fold, with 38, 78, and 11% recovery, respectively. Each purified component was serologically active, distinct, and free from the other components. The purified EF produced edema when mixed with PA, and the purified PA was an active immunogen. The components did not appear to be simple proteins by spectrophotometric analysis. As they were purified, the pH range in which they were most stable narrowed, centering between pH 7.4 and 7.8. Heat readily destroyed the biological activity of the components but not their serological activity. The rat lethality test showed that, with a constant amount of LF and an increasing amount of PA, the time to death reached a minimum and then was extended. When an increasing amount of LF was added to a constant amount of PA, the time to death became shorter as more LF was added. The biological, immunological, and serological properties of the components were shown to vary independently with storage and extent of purification so that serological activity was not always directly correlated with biological activity. Evidence is presented that the components can exist in different molecular configurations or as aggregates, and that this property is influenced by the state of component purity and by the environment.     

Antibiotic Treatment of Anthrax Infection in Mice

Quantitative measurements of mean time to death, percentage of survivors, and viable cell populations in the whole body were employed to determine the effects of penicillin, dihydrostreptomycin, chlortetracycline, oxytetracycline, chloramphenicol, and antiserum on the course of anthrax infection in mice. By all parameters tested, penicillin and dihydrostreptomycin were most effective in the treatment of the disease. Therapy initiated in the later stages of the disease was more effective than that initiated in the earlier stages. Quantitative studies indicated that it was more difficult to eliminate organisms from the kidney than from any other organ or tissue. These measurements for the evaluation of antibiotic therapy are suggested for the study of other bacterial diseases.     

A continuous, farm-scale, solid-phase fermentation process for fuel ethanol and protein feed production from fodder beets

Fuel ethanol (95%) was produced from fodder beets in two farm-scale processes. In the first process, involving conventional submerged fermentation of the fodder beets in a mash, ethanol and a feed (PF) rich in protein, fat, and fiber were produced. Ethanol yields of 70 L/metric ton (7 gal/ton) were obtained; however, resulting beers had low ethanol concentrations [3-5% (v/v)]. The high viscosity of medium and low sugar, beet mashes caused mixing problems which prevented any further increase of beet sugar in the mash. The severely limited the maximum attainable ethanol concentration during fermentation, thereby making the beer costly to distill into fuel ethanol and the process energy inefficient. In order to achieve distillably worthwhile ethanol concentrations of 8-10% (v/v), we developed and tested a solid-phase fermentation process (continuous). In preliminary trials, this system produced fermented pulp with over 8% (v/v) ethanol corresponding to an ethanol yield of 87 L/metric ton (21 gal/ton). Production costs with this novel process are $0.47/L ($1.77/gal) and the energy balance is 2.11. These preliminary cost estimates indicate that fodder beets are potentially competitive with corn as an ethanol feedstock. Additional research, however, is warranted to more precisely refine individual costs, energy balances and the actual value of the PF.     

Molecular analysis of a case of meiotic recombination leading to cri-du-chat syndrome

This paper describes a molecular investigation of a woman with an apparent large pericentric inversion of chromosome 5, inv(5)(p14;q35), and one normal chromosome 5 and her child, who was born with cri-du-chat syndrome. The four chromosome 5 homologs from the proband and his mother were isolated in somatic cell hybrids, and their haplotypes were determined at nine loci polymorphic for restriction enzyme sites. The deleted chromosome in the proband was shown to carry alleles from both maternal homologs, verifying molecularly that a meiotic recombination event in the mother gave rise to her son's deleted chromosome 5. The single crossover was presumably near the centromere.     

Preservation of ATP in Hypersaline Environments

High concentrations of particulate ATP were found in the anoxic brines of the Orca Basin and East Flower Garden, Gulf of Mexico. Other measurements indicative of growth and respiration suggested that the microbial community in the brines was inactive, but somehow the ATP associated with the cells persisted. Conceivably, when cells growing just above the interface sank into the brine, the increased osmotic stress could elicit an osmoregulatory response resulting in increased ATP. It was also possible that hydrolytic enzymes were inactivated, resulting in the preservation of ATP. Experiments in which a culture of marine bacteria was suspended in menstrua of different salinities comparable to those found across the Orca Basin interface revealed that as salinity increased, ATP increased three- to sixfold. Within 24 h the ATP fell to its initial level and remained at that concentration for 3 days, at which time the experiment was terminated. In contrast, the control suspensions, at a salinity of 28% (grams per liter) had 1/10th of the initial ATP concentration when the experiment was ended. Cells were also exposed to killing UV irradiation, enabling us to demonstrate with absolute certainty that cellular ATP could be preserved. At the end of the experiment, the viable component of the population was reduced by orders of magnitude by UV irradiation, but the ATP levels of the cells suspended in brine did not decrease. In certain environments it appears that the conventional analytical tools of the microbial ecologist must be interpreted with caution.     

Monoclonal antibodies specific to apical surfaces of rat alveolar type I cells bind to surfaces of cultured, but not freshly isolated, type II cells

The alveolar surface of the lung is lined by two classes of epithelial cells, type I and type II cells. Type I cells cover more than 97% of the alveolar surface. Although this cell type is felt to be essential for normal gas exchange, neither unique identifying characteristics nor functions have been described for the type I cell. We have produced monoclonal antibodies to (a) component(s) of molecular weight 40,000 and 42,000 of the apical surface of rat alveolar type I cells. The antibodies are specific to the lung in Western blots of organ homogenates. In immunocytochemical studies of frozen lung at the level of both light and electron microscopy, the monoclonal antibodies appear to react specifically with the apical plasma membrane of type I cells. Airway, vascular, interstitial cells, type II cells and macrophages are not immunoreactive. Western blots of isolated type I cells (approx. 70% pure) also demonstrate immunoreactivity at molecular weights of 40,000 and 42,000. When the lung is injured, type I cells may be damaged and sloughed from the alveolar surface. Alveolar repair occurs when the second type of alveolar cell, the type II cell, divides. Cell progeny may retain type II cell morphology or may differentiate into type I cells. Western blots of freshly isolated type II cells (approx. 85% pure) do not display immunoreactivity with our monoclonal antibodies. However, type II cells maintained in culture acquire immunoreactivity to monoclonal antibodies, demonstrating that type II cells in vitro have the capacity to develop a characteristic associated with type I cells in situ. The availability of markers for a specific membrane component of type I cells should facilitate the study of many questions on alveolar functions, development and response to injury.     

A two-stage classifier for identification of protein-protein interface residues

MOTIVATION: The ability to identify protein-protein interaction sites and to detect specific amino acid residues that contribute to the specificity and affinity of protein interactions has important implications for problems ranging from rational drug design to analysis of metabolic and signal transduction networks. RESULTS: We have developed a two-stage method consisting of a support vector machine (SVM) and a Bayesian classifier for predicting surface residues of a protein that participate in protein-protein interactions. This approach exploits the fact that interface residues tend to form clusters in the primary amino acid sequence. Our results show that the proposed two-stage classifier outperforms previously published sequence-based methods for predicting interface residues. We also present results obtained using the two-stage classifier on an independent test set of seven CAPRI (Critical Assessment of PRedicted Interactions) targets. The success of the predictions is validated by examining the predictions in the context of the three-dimensional structures of protein complexes.     

Arrivals of Hitchhiking Insect Pests on International Cargo Aircraft at Miami International Airport

In a study of hitchhiking or contaminating insect pests on international cargo aircraft at Miami International Airport from 1998 to 1999, it was found that contamination rates were greatest, 23%, on cargo flights from Central America and much lower, near 5%, on flights from all other regions. We reanalyzed the study data to test for associations between contaminated flights and factors such as season, cargo type, and time of departure (night or day), and developed probabilistic models for predicting insect pest arrivals by region and pest risk levels. Significant (P < 0.05) associations were detected between contaminated flights and (1) wet season flights from Central America, (2) flights carrying plant products and clothing or fabrics, and (3) flights departing at night from the country of origin. In Monte Carlo simulations, numbers of arriving mated insect pests were greatest for cargo flights from Central America, because of great contamination rates, and South America, because of the large volume of flights from there. Few insects arrived on flights from the Caribbean, and few high-risk insects arrived from anywhere. Although the likelihood of establishment in South Florida via this pathway could not be estimated, based upon arrivals the greatest threats were posed by moderate-risk insect pests on flights from Central and South America. Simulations indicated that switching to daytime departures only reduced pest arrivals by one-third. The simplest mechanism for pathway entry that explains the associations found is that insects entered aircraft randomly but sometimes remained because of the presence of certain cargo types. Hence, contamination rates were greater during the wet season because of greater abundance locally, and on nighttime flights because of greater abundance around lighted loading operations. Empty planes probably had no pests because pests had no access to holds. Thus, the best mitigation strategies for this pathway will likely be those that exclude insects from holds or reduce the attractiveness of night loading operations. Optimizing inspections based on associations is also possible but will be less effective for regions such as South America, with high flight volumes and low contamination rates. Comparisons to other pathways indicates the potential importance of hitchhikers on cargo aircraft at MIA.     

基于最大熵生态位模型的中华穿山甲潜在适宜生境预测

中华穿山甲（Manis pentadactyla）属于全球极度濒危物种,也是我国一级保护动物。对中华穿山甲的非法捕杀曾导致其种群数量锐减。但是,近年来相关研究报道较少,穿山甲分布状况不明,极大地制约了对该物种的有效保护。搜集了近年来国内中华穿山甲的救护记录和救护新闻,甄别出67个记录分布点,利用最大熵模型软件（MaxEnt）进行因子筛选,结果表明最冷季度降水量、人口密度、年降水量、坡度、坡向、海拔等6个环境变量是与中华穿山甲分布显著相关的影响因子。基于6个主导环境变量构建的MaxEnt模型AUC平均值为0.961±0.014,预测结果达到极好标准。刀切法（Jackknife）表明,其中最冷季度降水量、年降水量、人口密度和海拔是影响中华穿山甲分布的主要因素。中华穿山甲适宜生境（出现概率大于0.498）具有以下特点：最冷季度降水量141.22-439.46 mm,年降水量1471.67-2386.56 mm,人口密度≥390人/km²,海拔<316.98 m。该模型预测中华穿山甲在我国的潜在分布适宜区主要位于我国长江以南地区,总面积约为74.27×10⁴ km²,占国土面积的7.73%,主要集中在江西、广东、湖南和广西省,面积分别占该区域的97.58%,89.65%,76.90%和73.08%;其次是浙江、福建、台湾和安徽省。湖北、江苏、四川、云南、贵州等省份也有中华穿山甲的零星分布。湖北东南部、江苏南部、浙江西南部和福建西北部等与江西接壤的区域也是中华穿山甲的重要潜在分布适宜区。明确中华穿山甲的潜在分布适宜区,可为该物种的种群保护和栖息地管理提供科技支撑。