Protein modification by a Maillard reaction intermediate methylglyoxal. Immunochemical detection of fluorescent 5-methylimidazolone derivatives in vivo

Methylglyoxal (MG), an endogenous metabolite that increases in diabetes, is a common intermediate in nonenzymatic glycation (Maillard reaction) in vivo. Here we describe the immunochemical approach to the detection of MG adducts in proteins in vitro and in atherosclerotic lesions of human aorta in vivo. The reaction of protein (bovine serum albumin) with MG led to selective loss of arginine and lysine residues, accompanied by the formation of 5-methylimidazolone (N delta-(5-methylimidazolon-2-yl)ornithine) and imidazolysine (1,3-di-lysino-4-methylimidazole) derivatives, respectively. The anti-5-methylimidazolone antibody was prepared by immunizing rabbits with a MG-keyhole limpet hemocyanin conjugate and purifying the serum on an affinity gel prepared by covalent attachment of the 5-methylimidazolone derivative. The antibody cross-reacted with the proteins treated with not only MG but trioses, such as hydroxyacetone, dihydroxyacetone, and glyceraldehyde. The immunohistochemical analysis revealed that atherosclerotic lesions of human aorta contained 5-methylimidazolone derivatives whose distributions were identical to those of advanced glycation end products (AGEs) detected by the anti-AGE antibody.     

Maximum likelihood analysis of algorithms and data structures

We present a new approach for an average-case analysis of algorithms and data structures that supports a non-uniform distribution of the inputs and is based on the maximum likelihood training of stochastic grammars. The approach is exemplified by an analysis of the expected size of binary tries as well as by three sorting algorithms and it is compared to the known results that were obtained by traditional techniques. Investigating traditional settings like the random permutation model, we rediscover well-known results formerly derived by pure analytic methods; changing to biased data yields original results. All but one step of our analysis can be automated on top of a computer-algebra system. Thus our new approach can reduce the effort required for an average-case analysis, allowing for the consideration of realistic input distributions with unknown distribution functions at the same time. As a by-product, our approach yields an easy way to generate random combinatorial objects according to various probability distributions.     

System specification experiments on a common benchmark

To contribute to the current effort toward the definition of a system-level design language, this article proposes a benchmark for system specification. The authors analyze the results of experiments using the benchmark with several languages     

Percutaneous bile duct drainage; experiences with a new type of endoprosthesis]

Self expandable stents were placed percutaneously in 105 patients with malignant biliary obstruction. Stent diameter was 1 cm; length, 3.5-10.5 cm. Of the 60 patients with common bile duct obstruction, 50 died 0.2-12 months (median 3 months) after stent insertion. Two patients developed recurrent jaundice and cholangitis after 6 and 12 months, respectively. One patient underwent reintervention. Ten patients, one after a successful reintervention, were alive without jaundice 1-8 months (median 5 months) after stent placement. Of the 45 patients with hilar lesions, 26 died 0.7-18 months (median 5 months) after stent placement, five of them with signs of cholangitis. Nineteen are alive 1-21 months (median 7 months) afterwards. Reinterventions were carried out in 13 patients (29%). The most common cause of stent malfunction was tumour overgrowth. Stent-related complications were seen in three patients.     

Symbolic pole/zero calculation using SANTAFE

The aim of symbolic analysis is to gain insight into circuit behavior. To study the behavior of analog circuits, the locations of the poles and zeros have to be known. Unfortunately, no general method exists to calculate the poles and zeros symbolically for polynomials of degree greater than four from transfer functions in coefficient form. The CAD tool SANTAFE (Symbolic Analysis of Transfer Functions) applies the signal-flow graph method, which permits to keep the result in a factorized or partially factorized form. The graphic view provided by a signal-flow graph offers insight into the internal interactions between the circuit elements and, as will be demonstrated, enables the user to perform circuit knowledge-based approximations. A novel procedure based on symbolic Newton-iteration, accurately calculates high-order transfer functions in the desired pole/zero form. Another special routine, based on element weight ratios rather than numerical values, enables the simplification of large symbolic expressions without numerical values for each parameter. With the program SANTAFE, even large networks can be analyzed symbolically. This will be shown with an example of a wide band BiCMOS operational amplifier     

Primary lung cancer in patients under 40 years of age

There seems to be a resistance of patients and physicians towards aggressive diagnostic evaluation of the symptoms of lung cancer in young people. We here review nine series of young patients with primary lung cancer. Patients below 40 years of age represent between 1.2 and 5% of the total lung cancer population. The distribution of sex and histopathologic findings is different, there being more women, fewer cases of squamous cell and more cases of small anaplastic and adenocarcinoma in the young group. Between 87 and 96% are smokers. There is a delay from the debut of symptoms to the first contact with a general physician of 2.4 to 10.8 months. There is a wide variation concerning tendency to operate with a frequency of curative resection of between 15 and 57%. Based on the survival of young patients who are treated by curative surgical resection, the outcome of surgical treatment for young patients does not differ from the general experience concerning resection in patients of all ages. Young patients who are found inoperable have worse survival than the older patients. Seventy to 90%, more than in the group of patients of all ages, have stadium II or III at the time of diagnosis. In conclusion, physicians should be aggressive with respect to the diagnostic evaluation even of young patients with symptoms suggestive of lung cancer.     

Pregnancy rates after timed AI of heifers following removal of intravaginal progesterone inserts

Reproductive performance of dairy heifers was compared for each of 2 synchronization protocols: The first group of 54 heifers was synchronized using intravaginal progesterone inserts (CIDR) plus estradiol cypionate (ECP) on d 0, PGF(2alpha) on d 7, and ECP again on d 8 (CIDR-ECP); a second group of 56 heifers was synchronized using CIDR and ECP on d 0, PGF(2alpha) on d 7, and GnRH on d 9 (CIDR-GnRH). All heifers received timed artificial insemination (TAI) at 48, 56, or 72 h after CIDR removal on d 7. Pregnancy diagnosis was conducted by ultrasonography 32 +/- 1 d post AI to confirm pregnancy and at 60 +/- 1 d post AI to determine embryo survival. Ovaries were monitored by ultrasonography daily from d 0 to 7 and twice daily from d 8 to ovulation to examine emergence of a new wave of follicles, size of the ovulatory follicle, and timing of ovulation on 15 heifers per protocol. New follicular development was detected 3.7 +/- 0.2 d after CIDR insertion. Heifers receiving CIDR-ECP had a shorter interval from CIDR removal to ovulation than heifers receiving CIDR-GnRH (63.8 +/- 3.0 vs. 71.6 +/- 2.3 h, respectively); however, ovulation occurred 39.8 +/- 3.0 h after ECP or 23.6 +/- 2.3 h after GnRH. Diameters of ovulatory follicles did not differ between treatments. Overall pregnancy rate for synchronized heifers was 60.1%, and embryo survival was 98%. Pregnancy rate for heifers synchronized with CIDR-ECP was 63.0% and similar to that in heifers synchronized with CIDR-GnRH (57.1%). Pregnancy rate was affected by time of AI for heifers synchronized using CIDR-ECP but not for those synchronized with CIDR-GnRH. Heifers in the CIDR-ECP group that were inseminated 56 h after CIDR removal had a higher pregnancy rate (81.0%) compared with heifers inseminated 48 (66.7%) or 72 h (50.0%) after CIDR removal. Either ECP or GnRH used in a CIDR-based TAI program in dairy heifers can achieve acceptable reproductive performance.     

Comparison of on-farm laboratory milk progesterone assays for identifying errors in detection of estrus and diagnosis of pregnancy

A field study was conducted in 10 commercial dairy herds to test the efficacy of a rapid enzyme-linked immunosorbent assay to identify errors in detection of estrus and diagnosis of pregnancy. Milk samples were collected at the milking following detection of estrus and on d 21 postinsemination. Progesterone was measured on-farm by an enzyme-linked immunosorbent assay and in the laboratory by radioimmunoassay. Agreement between the two methods of progesterone analysis (n = 820) was 89.8%. There were 93 cows (18.7%) by laboratory analysis and 126 cows (25.4%) by on-farm analysis for which concentration of progesterone in the sample collected at suspected estrus (n = 497) was higher than normal estrual values. Exclusion of these cases increased the overall accuracy of pregnancy diagnosis by 6.8 and 6.4% for laboratory and on-farm methods. Overall agreement between pregnancy diagnosis by milk progesterone at d 21 postinsemination and diagnosis by return to estrus or palpation of reproductive organs was 80.5 and 75.8% for laboratory and on-farm methods (75.0% and 69.7% for pregnant cows and 94.5 and 93.9% for nonpregnant cows). These results indicate that an enzyme-linked immunosorbent assay for milk progesterone performed on-farm was equal in accuracy to a laboratory radioimmunoassay for identifying errors in detection of estrus and diagnosis of pregnancy.     

A GC–MS-based metabonomic investigation of blood serum from irritable bowel syndrome patients undergoing intervention with acidified milk products

In the present study, the use of gas chromatography mass spectrometry (GC–MS)-based metabonomics to characterize blood serum in an intervention study of patients suffering from the common gastrointestinal disorder irritable bowel syndrome (IBS) was investigated. The patients included in the study consumed an acidified milk product with (n = 30) or without probiotics (n = 31) (Lactobacillus paracasei F19, Lactobacillus acidophilus LA-5 and Bifidobacterium lactis BB-12) for an 8-week period, and blood serum samples were collected before and after the intervention. Acidified milk is commonly used as a delivering vector for probiotics in commercial consumer settings. The serum samples were extracted and derivatized using N-Methyl-N-(trimethylsilyl) trifluoroacetamide (MSTFA), and GC–MS analysis was carried out. Multivariate data analysis including principal component analysis (PCA), orthogonal partial least squares-discriminant analysis (OPLS-DA), and S-plot was applied on the obtained GC–MS data, which revealed higher serum lactate, glutamine, proline creatinine/creatine, and aspartic acid levels and lower serum glucose levels after the intervention period for both treatment groups. Consequently, the present study indicated an effect of acidified milk consumption on the plasma metabolite profile, which was independent of a concomitant intake of probiotics. In addition, the present study demonstrates that GC–MS is a useful analytical technique for metabonomics studies of blood serum.