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Bacterial contamination of nylon corneal sutures.

We report the findings of a prospective study into the bacterial contamination of monofilament nylon sutures removed from corneal wounds following cataract surgery. Sutures were classified as tight, loose or broken at the time of removal. Loose and broken sutures showed significantly more bacterial contamination than tight sutures (p < 0.001, chi squared). Positive cultures were obtained from 2 (6.2%) of 32 tight sutures, 14 (38.9%) of 36 loose sutures and 11 (37.9%) of 29 broken sutures. Staphylococcus epidermidis was the most commonly isolated organism (isolated in pure growth from 22 (81.5%) of 27 positive cultures). These findings may explain the occasional association of biodegraded corneal monofilament nylon sutures and suppurative keratitis and highlight the potential risk of seeding a suture track infection at the time of suture removal. They also emphasise the need for prophylactic topical antibiotic when removing biodegraded sutures.

Bacteria↗

Large scale purification of human blood CD34+ cells from cryopreserved peripheral blood stem cells, using a nylon-fiber syringe system and immunomagnetic microspheres.

Isolation of large numbers of human peripheral blood CD34+ cells could lead to therapeutic applications, including purging of malignant cells from blood cell transplantations, purging of T cells from allogeneic bone marrow, and even blood cell transplantation. This procedure has limitations if there are not sufficient numbers of progenitor cells in the leukapheresis concentrates available for selection after detection of tumor cells in apheresis products. Use of frozen/thawed peripheral blood mononuclear cell (PBMC) samples would make feasible pooling of two or even more stem cell harvests collected at different time points and the total number of CD34+ progenitor cells available would increase. We established an efficient method for purification of CD34+ cells from cryopreserved apheresis products, using a nylon-fiber syringe system and immunomagnetic microspheres. We compared purity, recovery rate and clonogenicity of CD34+ cells purified from fresh (n = 22) and cryopreserved apheresis products (n = 14), using a nylon-fiber syringe system and immunomagnetic microspheres. The purity of CD34+ cells from cryopreserved products was less than that from fresh products (85.9 +/- 14.4% vs 94.6 +/- 10.0%), but the recovery rate of CD34+ cells and colony-forming cells was comparable between fresh and cryopreserved products. One patient underwent grafting with peripheral blood CD34+ cells selected after freezing, with good success. Therefore, these cells are capable of rapidly reconstituting hematopoiesis after high-dose chemotherapy. Bone Marrow Transplantation (2000) 26, 787-793.

Antigens, CD34↗

No stop codons in the antisense strands of the genes for nylon oligomer degradation.

Genes for nylon oligomer-degrading enzymes are unique in the sense that the enzymes encoded by them are found not to have any appropriate substrates during most of the period of their evolution. Furthermore, these nylB genes form a family not related to any other known gene families. The base sequences of these genes were examined and a common characteristic was found: a long stretch of sequence without chain-terminating base triplets, defined as a nonstop frame (NSF), is being maintained on the antisense strand. Moreover, a certain coding frame is open for both the sense and the antisense sequences, while the other frames have many stop codons. The probability of the presence of these NSFs on the antisense strand of a gene is very small (0.0001-0.0018). In addition, another gene for nylon oligomer degradation was found to have a NSF on its antisense strand, and this gene is phylogenically independent of the nylB genes. Therefore, the presence of these NSFs is very rare and improbable. Even if the common ancestral gene of the nylB family was originally endowed with a NSF on its antisense strand, the probability of this original NSF persisting in one of its descendants of today is only 0.007. Unless an unknown force was maintaining the NSF, it would have quickly disappeared by random emergences of chain terminators. Therefore, the presence of such rare NSFs on all three antisense strands of the nylB gene family suggests that there is some special mechanism for protecting these NSFs from mutations that generate the stop codons. Such a mechanism may enable NSFs to evolve into new functional genes and hence seems to be a basic mechanism for the birth of new enzymes.

Amidohydrolases↗

Microbially corrected amino acid composition of rumen-undegraded feed protein and amino acid degradability in the rumen of feeds enclosed in nylon bags.

1. In the previous work (Varvikko & Lindberg, 1985), 15N-labelled rapeseed (Brassica napus), barley, ryegrass (Lolium perenne) and barley straw were incubated in the rumen in nylon bags for 5, 12 and 24 h and microbial nitrogen in the residues was quantified using the feed 15N-dilution method. In the present study, residual amino acids (AA) of these feeds were analysed, and microbially corrected AA of feed origin (feed AA) were estimated as the difference between total residual AA and respective microbial AA, assuming a constant AA composition for the microbial protein. 2. In barley and barley-straw residues, and also in ryegrass incubated in the rumen for 24 h, very large enrichment by microbial N and AA-N was found. The microbial enrichment was rather small in rapeseed residues and ryegrass incubated for 5 or 12 h. During the rumen incubation, feed N and AA-N (g/kg feed dry matter (DM] decreased very clearly in all the feeds, and feed and incubation time effects were always statistically significant (P less than 0.001). 3. The slow degradation of essential (E) feed AA compared with the respective non-essential (NE) AA degradation increased the proportion of feed EAA (g/kg determined feed AA) in barley and barley-straw residues. In rapeseed and ryegrass, residual feed EAA:NEAA remained very similar to the original. Branched-chain (Br) AA tended to increase proportionally in all the feed residues, suggesting these AA to be, on average, more resistant against microbial degradation in the rumen than other AA. Similarly, lysine was clearly increased in barley residues. A rumen degradation faster than the average rate caused decreased residual feed glutamic acid in rapeseed; methionine, alanine and glycine in barley; arginine and alanine in ryegrass; and methionine, asparagine and tyrosine in barley straw. Feed and incubation time effects were significant (P less than 0.05-0.001) for feed AA (g/kg determined feed AA) grouped as EAA, BrAA or NEAA, and for most individual AA, as well as for feed AA disappearance (%) and relative amounts (%) of feed AA in the respective residual AA. 4. According to present findings, AA composition of the rumen-degraded vegetable feed residues may markedly differ, either quantitatively or qualitatively (or both), from their original AA composition. When determining the feed AA composition of nylon-bag residues, the microbial error may be very large with starchy or fibrous feeds of low protein content. The microbial AA do not, however, considerably confuse the AA determination of protein-rich feeds.

Amino Acids↗

Analysis of migrants from nylon 6 packaging films into boiling water.

Ultra-violet spectrophotometry (UV), high performance liquid chromatography (HPLC) and liquid chromatography coupled to mass spectroscopy (LC-MS) were used to identify and quantify oligomers extracted with boiling water from two different nylon 6 films used in boil-in-bag food packaging. The results indicated the loss of up to 1.5% of the original nylon film weight, into the boiling water, as caprolactam and cyclic oligomers up to the nonamer. Extraction time, thickness and type of film used, were found to be parameters which affected the levels of these migrants. These results will be relevant to situations in which food is cooked in the water used to heat the pouch contents.

Caprolactam↗

Nylon adherent antigen-specific rosette-forming T cells.

Shortly after intravenous immunization of mice with heterologous erythrocytes (RBC) antigen-specific Thy 1+ cells which form rosettes with the immunizing RBC (thymic-derived lymphocytes-forming rosettes [T-RFC]) appear in the spleen. These T-RFC are much less stable than Thy 1- RFC (non-thymic-derived [B-RFC]) although most if not all of both classes of RFC adhere to nylon. T-RFC are induced with low doses of antigen (which fail to induce B-RFC) and are inhibited by higher antigen doses which are optimal for induction of B-RFC. Pretreatment of mice with cyclophosphamide prevents the high dose inhibition of T-RFC. Although there are many parallels between the production of T-RFC and delayed-type hypersensitivity (DTH) it is unlikely that the T-RFC are essential for DTH reactions since DTH can be transferred with cells which pass through nylon, and such cells are almost totally depleted of T-RFC. Thus immunization can lead to the production of large numbers of antigen-specific T-RFC whose functional role in the immune response is unknown. However, the characteristics of the T-RFC suggest that they may play an important role in amplification of suppressor cell activity.

Animals↗

Induction of human interferon gamma in nylon-column nonadherent human lymphocyte cells by heat-treated poly I:poly C.

Human peripheral blood mononuclear leukocytes (PBML) were separated by Ficoll-hypaque gradient from human peripheral whole blood cells. The PBML were further separated by nylon-column. When nylon-column nonadherent cells (NNA cells) were induced by poly I:poly C which was prepared by heat-treatment, high levels of acid- and heat-labile interferon (IFN) were obtained. The IFN was not neutralized with human IFN-alpha or -beta anti-serum, but was completely neutralized with human IFN-gamma anti-serum, indicating that the IFN was human IFN-gamma.

Cell Adhesion↗

A Simple Purification of Indole-3-Acetic Acid and Abscisic Acid for GC-SIM-MS Analysis by Microfiltration of Aqueous Samples through Nylon.

A simple procedure was developed for the partial purification of plant tissue samples to be analyzed simultaneously for indole-3-acetic acid (IAA) and abscisic acid (ABA). The procedure relies on removal of contaminants by filtration through nylon and partitioning into dichloromethane. This procedure successfully purified both IAA and ABA from muskmelon, cotton, and broccoli tissue. Twenty individual samples can be purified and methylated in 8 h for analysis of free IAA and ABA with gas chromatography-selected ion monitoring-mass spectrometry. The use of microfiltration of aqueous samples through nylon offers new opportunities for improving the efficiency of existing sample purification procedures.

Journal Article↗

Cross-species hybridisation of pig RNA to human nylon microarrays.

BACKGROUND: The objective of this research was to investigate the reproducibility of cross-species microarray hybridisation. Comparisons between same- and cross-species hybridisations were also made. Nine hybridisations between a single pig skeletal muscle RNA sample and three human cDNA nylon microarrays were completed. Three replicate hybridisations of two different amounts of pig RNA, and of human skeletal muscle RNA were completed on three additional microarrays. RESULTS: Reproducibility of microarray hybridisations of pig cDNA to human microarrays was high, as determined by Spearman and Pearson correlation coefficients and a Kappa statistic. Variability among replicate hybridisations was similar for human and pig data, indicating the reproducibility of results were not compromised in cross-species hybridisations. The concordance between data generated from hybridisations using pig and human skeletal muscle RNA was high, further supporting the use of human microarrays for the analysis of gene expression in the pig. No systematic effect of stripping and re-using nylon microarrays was found, and variability across microarrays was minimal. CONCLUSION: The majority of genes generated highly reproducible data in cross-species microarray hybridisations, although approximately 6% were identified as highly variable. Experimental designs that include at least three replicate hybridisations for each experimental treatment will enable the variability of individual genes to be considered appropriately. The use of cross-species microarray analysis looks promising. However, additional validation is needed to determine the specificity of cross-species hybridisations, and the validity of results.

Journal Article↗

Phosphorescence properties of p-aminobenzoic acid immobilized on a nylon membrane.

The properties of nylon as a new material for obtaining room-temperature phosphorescence from p-aminobenzoic acid (PABA) have been investigated. Although PABA shows native phosphorescence when adsorbed on a nylon membrane, a significant improvement in both sensitivity and limit of detection is achieved in the presence of alpha-cyclodextrin and heavy atoms. An additional enhancement of the phosphorescence signal is attained when the measurements are carried out under a nitrogen atmosphere. The analytical figures of merit obtained under the best experimental conditions are: linear calibration range from 0.6 to 6 ng/spot (the lowest value corresponds to the quantitation limit); correlation coefficient, 0.998 for 18 data points; relative standard deviation, 2.2% (n = 5) at a level of 2.4 ng/spot; and limit of detection, 0.2 ng/spot (calculated according to Clayton's definition).

Journal Article↗

Methods for gene expression profiling in dermatology research using DermArray nylon filter DNA microarrays.

Here we present methods of gene expression profiling using nylon filter deoxyribonucleic acid (DNA) microarrays and radiolabeled and nonradiolabeled hybridization probes. DermArray(R) nylon filter DNA microarrays were designed specifically for use in dermatology research. A patent-pending method was used to select approx 4400 highly informative, sequence-verified human cDNA clones for this DNA micro array. Using DermArray(R) filters, biomarkers have been discovered for normal and pathologic cells from skin, and for responses to dermatologic drugs. As an example, gene expression profiling was performed with hydroquinone-treated SKMel-28 cells, a melanoma cell line. Also included are the methods for bioinformatic analysis using Pathwaystrade mark software.

Computational Biology↗

Robust and sensitive nylon hybridization membrane suitable for high-throughput robotic arraying applications.

An important aspect of automated macroarraying is the suitability of the nylon membrane selected on which samples are to be arrayed. PerForma is a positively charged nylon membrane that has been developed specificallyfor automated macroarraying. Tests usingfluorescent hybridization detection methods have shown that immobilized DNA amounts as low as 0.25 pg can be detected and that positive signals are obtainable after 21 stripping cycles. This report describes the improved colony growth, improved handling characteristics, increased hybridization detection sensitivity, and increased stripping and reprobing capability obtained using PerForma.

Cells, Cultured↗

Immobilon-Ny+ nucleic acid blotting membrane: an advanced nylon membrane optimized for superior fixation and reprobing.

The use of charged nylon membranes in nucleic acid blotting applications has become an important factor in the success of hybridization-based assays. Retention of nucleic acids on these membranes is promoted by baking at 80 degrees C under vacuum or by exposure to short wavelength UV light, with the latter method preferred. Immobilon-Ny+ is an advanced, positively charged nylon membrane that has been optimized to show superior retention of target DNA and RNA under hybridization conditions. Higher signal levels are obtained in these assays compared to competitive membranes, even after 13 cycles of probing. This report illustrates the superior performance of Immobilon-Ny+ in 32P and chemiluminescent hybridization assays on blotted DNA and RNA.

Blotting, Northern↗

Evaluation of mobile nylon bag technique for determining apparent ileal digestibilities of protein and amino acids in growing pigs.

The mobile nylon bag technique (MNBT) may offer a simple, rapid means for assessing ileal AA digestibility of pig feed ingredients. In the present study, the effects of washing bags recovered from digesta, the amount and fineness of feeds, and feed trypsin inhibitor activity on apparent ileal digestibilities (AID) of CP and AA were determined with the MNBT. Twenty-four ileorectal anastomosed pigs (Yorkshire x Chinese Black barrows, 30 kg initial BW), of which 12 were fitted with duodenal T-cannulas, were used. Not washing the bags recovered from ileal digesta resulted in a reduction (P < 0.05) in apparent ileal digestibilities of CP and AA determined by MNBT. Washing the bags for more than 4 min overestimated (P < 0.05) the apparent ileal digestibilities of CP and AA compared with those determined with the anastomosed pigs. Sample size and fineness of grinding also affected (P < 0.05) apparent ileal digestibilities of CP determined by MNBT. The apparent ileal digestibilities of CP determined by MNBT were reduced (P < 0.05) when sample size exceeded 0.75 g and when feed was ground through screens with a mesh size of more than 1.0 mm. The closest agreement between results obtained by MNBT and a conventional ileal digestibility assay occurred when 0.75 g of feed ground through a 1.0-mm mesh screen was used per bag and bags were washed for 2 min after retrieval from digesta. Further studies are warranted to investigate the use of the mobile nylon bag technique for predicting the ileal digestibilty of AA for feeds containing antinutritional factors.

Amino Acids↗

The evaluation of the mobile nylon bag technique for determining the apparent protein digestibility in a wide variety of feedstuffs for pigs.

Studies were conducted with four (50 to 70 kg) barrows to evaluate the mobile nylon bag technique (MNBT) for determining the apparent protein digestibility (AD) in a variety of feedstuffs. One-gram samples of each feedstuff, ground through a 1.0-mm mesh screen, were enclosed in nylon bags (25 X 40 mm; 48 microns mesh). Following predigestion (.01 N HCl; 4,000 Folin Intestinal Pepsin Units/liter), four bags of each sample were inserted into each pig via a duodenal T-cannula. The indigested contents were pooled within pig and feedstuff prior to crude protein analysis. The AD of each feedstuff was determined previously with four barrows according to the conventional method (CM), based on a 5-d total collection of feces following a 14-d adaptation period. The following results, AD (%), were obtained with the MNBT (n = 4) and CM (n = 4), respectively. Barley, 70.1 vs 81.2 (P less than .05); corn, 72.3 vs 82.1 (P less than .05); wheat, 79.6 vs 86.4 (P less than .05); corn gluten feed, 73.9 vs 70.9; rice bran, 57.3 vs 57.4; wheat middlings, 70.8 vs 79.4 (P less than .05); soybean meal, 89.9 vs 93.0; a mixture of soybean meal and hulls, 86.9 vs 88.8; coconut expeller, 65.0 vs 69.8; fababeans, 79.5 vs 78.5; peas, 87.6 vs 88.1; meat meal, 83.2 vs 84.0; complete diet (4.3% crude fiber), 76.3 vs 83.6 (P less than .05); and complete diet (13.6% crude fiber), 56.5 vs 55.0.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Technical note: assessment of recovery site of mobile nylon bags for measuring ileal digestibility of starch in dairy cows.

The objective of this study was to evaluate recovery site of mobile nylon bags for measuring ileal digestibility of ruminally undegraded starch in dairy cows. Eight feed samples of untreated and treated concentrates were examined. Three lactating cows equipped with rumen fistula and duodenal and ileal cannulas were used in the experiment. The mobile nylon bags containing intact feeds or residues after a 12-h ruminal incubation were pretreated using a 2-step procedure to simulate abomasal digestion before insertion through the duodenal cannula. To assess the effect of hindgut fermentation on starch digestibility, approximately half of the bags were collected from the ileum and half from the feces. The results indicate that feed samples should be preincubated in rumen before insertion into duodenum, and that samples with relatively high fractions of rumen-undigestible starch should be collected from the ileum instead of from feces.

Animal Feed↗

The Spot Fitting and Expression Quantification for the Nylon Filter cDNA Microarray.

High-density cDNA microarray is important in monitoring large scalegenomic expression profile. With radiolabeled cDNA sample, nylon filter cDNA microarrays have high sensitivity and wide linear range of hybridization signals, so it can be employed to detect many important low-abundance cDNAs. However, the nylon filter tends to be randomly contaminated, and the array spots are prone to dispersion and saturation, resulting in inaccurate expression value. Based on aphysical model, we address a novel treatment with accurate positioning, complement of saturation and correction of interference. Thus the accurate quantificati on of expression can be extracted with remarkably improved reproducibility.

Journal Article↗

The involvement of polymorphonuclear leukocytes in the pathogenesis of bronchopneumonia in calves. V. Adherence to nylon fibres.

The adherence of neutrophils from bronchopneumonic calves to nylon fibres and the influence of this adherence on O2- and H2O2 production were studied. Polymorphonuclear leukocytes from bronchopneumonic calves were found to produce much more superoxide anion and hydrogen peroxide than neutrophils from healthy animals. A higher production of these compounds was observed in granulocytes adhering to nylon fibres than in cells in suspension. It is suggested that oxygen radicals production induced by the contact of granulocytes with a solid surface plays a role in the destruction of endothelium in vivo.

Animals↗