Evaluation of a real-time PCR assay quantifying the ruminal pool size and duodenal flow of protozoal nitrogen

We have recently developed a real-time polymerase chain reaction (PCR) assay to quantify copies of the genes encoding protozoal 18S rRNA. The assay includes procedures for isolating and concentrating protozoal cells from the rumen for use as a standard to convert 18S rRNA gene copies to a biomass basis. The current objectives were to 1) determine the degree of reduction of bacterial contamination in the protozoal standard, 2) determine if protozoal standards derived from ruminal fluid are appropriate for predicting duodenal flows, and 3) evaluate the assay's determined values for protozoal N in the rumen and flowing to the duodenum compared with independent measurements. Our protozoal collection method reduced non-associated bacterial contamination by 33-fold, the contamination of which could otherwise significantly bias RNA (microbial marker) and N percentages of concentrated protozoal fractions. Based on denaturing gradient gel electrophoresis, the use of protozoal cells isolated from ruminal fluid appears appropriate for use in quantitative assays determining protozoal N flow postruminally. Using real-time PCR, protozoal N was determined to be 4.8 and 12.7% of the rumen microbial N pool and 5.9 and 11.9% of the duodenal flow of microbial N on diets containing low (16%) or high (21%) forage neutral detergent fiber, respectively, which were comparable with independent measures and expectations.     

Assessment of iron bioavailability from twenty elite late‐maturing tropical maize varieties using an in vitro digestion/Caco‐2 cell model

An in vitro digestion/Caco‐2 cell model was used to assess iron bioavailability of twenty elite late‐maturing tropical maize varieties grown in three diverse agroecologies in West and Central Africa (WCA). Kernel‐iron concentration of the varieties, averaged across locations, varied from 19.2 to 24.4 mg kg^?1, while mean kernel‐zinc concentration ranged between 19.4 and 24.6 mg kg^?1. Significant differences in iron bioavailability were observed among varieties, but the environment had no significant effect. Mean bioavailable iron ranged between 14% below and 43% above the reference control variety, TZB‐SR. Variety DMR‐LSR‐Y with the highest concentrations of kernel‐iron and ‐zinc of 24–25 mg kg^?1 across the three locations had a similar quantity of bioavailable iron as the reference control, TZB‐SR. In the long run this variety could be potentially effective in reducing iron deficiency because of its high kernel‐iron. The most promising varieties were Mid‐altitude STR synthetic and ACR91SUWAN‐1‐SRC1. They had kernel‐iron and ‐zinc levels of 22–24 mg kg^?1 and bioavailable iron 24–36% higher than the reference control, TZB‐SR. Additional research is necessary to determine if the increases in kernel‐iron concentration and bioavailable iron observed in this study can significantly improve the iron status of individuals in WCA at risk for iron deficiency. Copyright © 2004 Society of Chemical Industry     

A simple method for on-chip, sub-femto Farad interconnectcapacitance measurement

In this letter, a sensitive and simple technique for parasitic interconnect capacitance measurement and extraction is presented. This on-chip technique is based upon an efficient test structure design that utilizes only two transistors in addition to the unknown interconnect capacitance to be characterized. No reference capacitor is needed. The measurement itself is also simple; only a dc current meter is required. Furthermore, the extraction methodology employs a self-checking algorithm to verify that the extracted capacitance value is consistent and accurate. The technique is demonstrated by extracting the capacitance of a single crossover between a Metal 1 line and a Metal 2 of 0.44 fF. The resolution limit is dominated by the matching of the minimum sized transistors used for the test structure. We estimate this resolution limit to be about 0.03 fF     

Variability in nanometer CMOS: Impact, analysis, and minimization

Variation is a significant concern in nanometer-scale CMOS due to manufacturing equipment being pushed to fundamental limits, particularly in lithography. In this paper, we review recent work in coping with variation, through both improved analysis and optimization. We describe techniques based on integrated circuit manufacturing, circuit design strategies, and mathematics and statistics. We then go on to discuss trends in this area, and a future technology outlook with an eye towards circuit and CAD-solutions to growing levels of variation in underlying device technologies.     

Long-term follow-up of an EORTC randomized prospective trial comparing intravesical bacille Calmette-Guérin-RIVM and mitomycin C in superficial bladder cancer. EORTC GU Group and the Dutch South East Cooperative Urological Group. European Organisation for Research and Treatment of Cancer Genito-Urinary Tract Cancer Collaborative Group

Posterior laryngeal clefts (PLCs) are described in the literature as rare laryngeal abnormalities. The authors believe type I clefts are much more common than previously reported. In two busy pediatric tertiary care centers, such clefts are the second most common congenital laryngeal finding at rigid endoscopy, second only to laryngomalacia. PLCs frequently present with symptomatology that can be attributed to other common disease processes and are often undiagnosed unless the surgeon maintains a high index of suspicion and specifically examines the posterior glottis by palpation during microlaryngoscopy. This report presents a series of 41 patients with type I PLCs, reviews their subtle and often confusing presenting signs and symptoms, and describes a simple yet reliable method of diagnosis.     

Surveillance for geographic and secular trends in congenital syphilis--United States, 1983-1991

PROBLEM/CONDITION: CDC monitors trends in the occurrence of congenital syphilis (CS) in the United States by using surveillance data sent from state and local health departments. Comparisons of data from this surveillance system with data from the Division of Sexually Transmitted Diseases/HIV Prevention and the Birth Defects Monitoring Program (BDMP) can be used to assess the potential effects of changes in case finding and reporting practices on these trends. REPORTING PERIOD COVERED: This report covers CS surveillance in the United States for the years 1983-1991. DESCRIPTION OF SYSTEM: Cases of CS among infants < 1 year of age and primary and secondary (P&S) syphilis among women are reported quarterly to CDC. The BDMP is a CDC national surveillance system that samples hospital discharge data on U.S. births. RESULTS: During the period 1983-1991, 12,151 CS cases were reported. Before 1988, regional CS incidence increased 35%-131% annually. Larger increases occurred in the Northeast (578%) in 1989 and in the South (178%), Midwest (244%), and West (777%) in 1990. Within regions, these larger increases were temporally related to increases in P&S syphilis in women and changes to a more sensitive CS case definition. INTERPRETATION: CS incidence has increased since 1983 in all regions of the United States. Increases since 1988 reflect both changes in surveillance reporting practices--the surveillance case definition for CS was changed in 1988 and further revised in 1989--and a true increase in incidence. ACTIONS TAKEN: These data indicate where CS prevention efforts need to be targeted. To facilitate reporting of CS cases, CDC has developed a) a shorter form for reporting cases of CS after 1991 and b) a software package for use by state and local health departments to enter and analyze CS data.     

Determination of rumen degradability and ruminal effects of three sources of methionine in lactating cows

Objectives were to quantify the ruminal effects and flows to the omasum of Met provided as 2-hydroxy-4-(methylthio)-butanoic acid (HMB), the isopropyl ester of HMB (HMBi), and DL-Met. Eight ruminally cannulated cows were used in a replicated 4 x 4 Latin square design. Treatments were 1) no Met (control), 2) HMB at 0.10% of DM, 3) HMBi at 0.13% of DM, and 4) DL-Met at 0.088% of DM. Diets were identical except for type of Met supplement and were based on corn silage and alfalfa hay at 30 and 13% of dietary DM, respectively. Samples of omasal fluid were used to determine the proportion of Met supplements passing out of the reticulorumen. Dry matter intake (20.1 kg/d) was restricted during the week of sampling to a maximum of 95% of ad libitum DMI determined during the first 2 wk of the period. Milk yields (37.7 +/- 0.8 kg/d) and fat concentration (3.42 +/- 0.15%) were not significantly different for control, HMB, HMBi, and DL-Met. Milk protein concentration (2.91, 2.95, 3.02, 2.96 +/- 0.07%, respectively) was significantly increased by the HMBi treatment. Rumen volatile fatty acids profile and NH3 concentrations were similar across treatments. Apparent ruminal digestibility of organic matter and neutral detergent fiber were higher for the three diets supplemented with Met sources than for the control diet. In situ rate of digestibility of CP from alfalfa hay, TMR, and corn silage was affected by Met sources. Passage rates of small particles (0.071/h) and fluid (0.167/h) were not affected by treatments. Protozoal counts in the rumen and omasum were not significantly affected by Met sources. Proportion of omasal N from bacterial N was not different (0.54 +/- 0.03), and bacterial N flow (305 +/- 24.4 g/d) was similar across treatments. The proportion of HMB that passed into the omasum was 5.3 +/- 1.5% of the amount consumed. Only a small amount (2.3%) of HMBi was found as HMB in the omasum. These results indicate that little HMB escapes ruminal degradation through passage to the omasum and that the site of HMBi absorption must be preomasal.     

Drag reduction in cocurrent horizontal natural gas-hexane pipe flow

Drag reduction in two-phase cocurrent horizontal natural gas-hexane pipe flow was studied experimentally. The effects of pipe diameter, gas flow rate, liquid flow rate, and additive concentration were investigated using the Dowell APE drag reducer. In the annular-mist flow regime studied, it was found that drag reduction increased with decreasing gas rate for a given liquid rate, drag reduction decreased with increasing friction velocity and it decreased with decreasing gas-liquid ratio for a given friction velocity. Although the drag reduction decreased rapidly as the liquid-gas ratio approached zero, drag reductions as high as 34 percent were obtained. Attempts to predict two-phase drag reduction were unsuccessful as it was found to depend upon friction velocity, liquid and gas rates, liquid-gas rate ratio, pipe diameter and additive concentration.