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PubMed2004

Field experiences in the control of avian influenza outbreaks in densely populated poultry areas.

From 1997 to 2003, Italy has been affected by two epidemics of highly pathogenic avian influenza (HPAI) and by several outbreaks of low pathogenic avian influenza (LPAI). In 1999-2000 a severe HPAI epidemic affected the country, causing 413 outbreaks: a total of about 16 million birds died or were stamped out. From August 2000 to March 2001, a H7N1 LPAI strain infected 78 poultry farms. The last affected flock was stamped out on the 26th of March 2001. In October 2002, another LPAI virus of the H7N3 subtype emerged and infected a total of 388 poultry holdings. Eradication measures were based on stamping out or controlled marketing of slaughtered birds on infected farms and on the prohibition of restocking. Restriction measures on the movement of live poultry, vehicles and staff were also imposed. To supplement these disease control measures, two emergency vaccination programmes, based on the "DIVA" (Differentiating Infected from Vaccinated Animals) strategy were implemented. The two vaccination campaigns (2000-2002 and 2002-2003) both resulted in the eradication of infection. However, the first campaign appeared to be more successful that the second and possible explanations are discussed.

Animals↗

PubMed1990

[Transcutaneous transport of trinitrin in children after application of nitroglycerin patches].

Plasma levels of nitroglycerin were measured 13 to 24 hours after the application of nitroglycerin stamps in 7 children aged from 1 month to 7 years and suffering from heart failure. The plasma levels varied considerably from one patient to another, confirming that the bioavailability of transdermal nitroglycerin is extremely variable in children. Only 3 patients had a nitraemia higher than 1 ng/ml, a figure compatible with a haemodynamic effect of the drug. No side-effect associated with the nitroglycerin stamps was observed. The dosage we use (a quarter of a stamp for 5 kg bodyweight) seems to be adequate at the beginning of treatment, to be progressively increased according to the way the drug is tolerated.

Administration, Cutaneous↗

PubMed1995

Including smoking status as a new vital sign: it works!

BACKGROUND: Despite the adverse health consequences of smoking, many physicians still neglect to counsel smokers to quit. This study evaluated the effect of including smoking status as a vital sign on the frequency of physician discussions with patients about smoking and physician advice to quit smoking. METHODS: A consecutive sample of adult ambulatory patients in our metropolitan family practice residency program completed exit surveys on physician and nurse counseling about smoking. Control group data were collected for 1 month before the change was made to include smoking status as a vital sign on patient charts. Charts were then marked with a stamp as a chart prompt in the vital signs section. Data were collected for 2 months after smoking status was added to the stamp. RESULTS: There were 637 individuals surveyed, of whom 179 were current smokers; 95 in the "prestamp" group and 84 in the "poststamp" group. The percentage of patient-physician encounters during which smoking was discussed increased from 47% to 86% (P < .001). Physician advice to quit increased from 50% to 80% (P < .001). Physician discussion of smoking with patients increased across all of the five stages of change but most dramatically (53% to 95%) in the "preparation" stage. Physicians were much less likely to counsel patients in the "precontemplation" stage to quit smoking. CONCLUSIONS: Including smoking as a new vital sign significantly increased the likelihood of smoking-related discussions between patients and their physicians. The stamp is inexpensive and easy to use, and because it is a one-time office system change, it is more likely to be implemented and maintained in busy practices.

Adult↗

PubMed1994

[3 different methods for evaluating Streptococcus mutans in the saliva].

Three different methods for detection and count of Streptococcus mutans in saliva have been compared with a microbiological method in order to estimate their efficacy and practicality. The study has been carried out on fifty children of six-eleven years old. The first method (stamp method) derives from that of Kohler and Bratthall, improved on the sampling, without saliva stimulation and, hence, more rapid. Furthermore, using a tongue depressor with a shovel-like extremity, we can either restrict the area where the grown colonies must be counted, avoiding any mistake, and stamp the spatula in the centre of the plate, not in the border, as the original method described, pressing it with the same force in every part, so that almost all the saliva collected can be plated on the Streptococcus mutans selective medium. As stated by this method, a tongue depressor is pressed first on the subject's tongue, then on a plate containing the medium. After incubation, the Streptococcus mutans colonies are counted on the plate's area limited by the stamp of the extremity of the spatula. According to Emilson we can also discriminate Streptococcus mutans and Streptococcus sobrinus colonies. The second method is a "strip mutans test" (Dentocult): saliva is stimulated making the subject chew a piece of paraffin and then it is collected passing a plastic spatula for ten times on the child's tongue. The spatula is introduced into a tube containing a liquid medium selective for Streptococcus mutans and then incubated.(ABSTRACT TRUNCATED AT 250 WORDS)

Bacteriological Techniques↗

PubMed1996

Experience with the modified Meek technique.

In 1958 Meek described the so called Meek-Wall dermatome to cut postage stamp skin grafts. This method was eclipsed by the introduction of mesh skin grafts. In 1993 Kreis and colleagues reintroduced a modified Meek technique using a dermatome running on compressed air. This technique has been used in our burn unit since August 1994. The aim of this paper is to compare the modified Meek technique with the mesh graft technique. Within a period of 20 months 41 patients were grafted using the modified Meek technique. The mean TBSAB was 54.4% with 50.0% full thickness burns. All patients were excised early. The expansion ratio was 1:4 and 1:6. In 20 patients the Meek technique was used exclusively for grafting of the trunk and the extremities with the exception of face, neck and hands. In 3 patients with a mean TBSAB of 68.3% a combination of postage stamp autologous skin grafts and cultured epithelial autografts (CEA) was applied. Compared with the mesh graft technique the Meek technique showed the following advantages: 1. The Meek method provides the true expansion ratio. 2. Small graft remnants can be utilized. 3. Grafting of full thickness burns up to 70 to 75% TBSAB becomes possible with one harvest of the donor sites. 4. The reliability of graft take is equal or better. 5. Epithelialization is achieved within 3 to 4 weeks depending on the expansion ratio. 6. The combination of widely expanded postage stamp split thickness grafts and CEA provides an excellent take rate and durable wound closure within a short time and avoids the problems associated with the engraftment of CEA on fascia. The method is simple but more demanding than the mesh technique. Compared with the mesh graft technique the preparation of Meek grafts is more time consuming and requires more staff than the Mesh technique. The cost of materials is higher. In our experience complete coverage of the Meek grafts with an overlay of meshed allografts after removal of the gauze as recommended by Kreis is not necessary using the 1:4 expansion ratio. Greater expansion ratios necessitate an overlay with meshed allografts. Regarding the scar formation no significant differences were observed compared with the mesh graft technique. In conclusion the modified Meek technique is reliable and simple to perform. This technique provides a sufficient expansion ratio enabling to graft patients with burns up to 75% TBSA with only one harvest of donor sides and without the necessity of CEA. In our opinion the Meek technique is reliable and simple to perform. This technique provides a sufficient expansion ratio enabling to graft patients with burns up to 75% TBSA with only one harvest of donor sides and without the necessity of CEA. In our opinion the Meek technique is advantageous in patients with burns greater than 45% TBSAB. In smaller burns mesh grafts should be used because of lower material cost and staff requirements. Especially in extensively burned patients the Meek technique may be cost effective avoiding the need of CEA.

Adolescent↗

PubMed2002

Microcontact printing on human tissue for retinal cell transplantation.

OBJECTIVES: To demonstrate that microcontact printing, a modern materials fabrication technique, can be used to engineer the surface of human tissue and to show that inhibitory molecules can be used to pattern the growth of retinal pigment epithelial cells or iris pigment epithelial cells on human lens capsule for transplantation. METHODS: Photolithographic techniques were used to fabricate photoresist-coated silicon substrates into molds. Poly(dimethylsiloxane)stamps for microcontact printing were made from these molds. The poly(dimethylsiloxane) stamps were then used to "wet-transfer" growth inhibitory molecules to the surface of prepared human lens capsules that were obtained during cataract surgery. Human retinal pigment epithelial and rabbit iris pigment epithelial cells were grown on a lens capsule substrate in the presence and absence of a patterned array of inhibitory factors. RESULTS: We found that human lens capsule could be microprinted with a precision similar to that obtained on glass or synthetic polymers. Retinal pigment epithelial cells and iris pigment epithelial cells cultured onto an untreated lens capsule showed spreading and formed into fusiform-appearing cells. In contrast, cells cultured on a lens capsule with a hexagonal micropattern of growth inhibitory molecules retained an epithelioid form within the inhibitory hexagons. CONCLUSION: Inhibitory growth molecules can be micropatterned onto human lens capsule, and these micropatterns can control the organization of retinal pigment epithelial cells or iris pigment epithelial cells cultured onto the lens capsule surface. CLINICAL RELEVANCE: Microprinting on autologous human tissue may facilitate efforts to effectively organize cell cultures and transplantations for the replacement of vital ocular tissues such as the retinal pigment epithelium in age-related macular degeneration.

Animals↗

PubMed2000

Enduring effects of nurse home visitation on maternal life course: a 3-year follow-up of a randomized trial.

CONTEXT: A home visitation program using nurses to improve maternal and child outcomes had favorable results in a randomized trial with a primarily white, semirural population. Many of the short-term findings have been replicated with urban blacks, but whether the program will continue to demonstrate effectiveness after its conclusion is uncertain. OBJECTIVE: To determine the effectiveness of a prenatal and infancy home visitation program on the maternal life course of women in an urban environment 3 years after the program ended. DESIGN AND SETTING: Three-year follow-up of a randomized controlled trial of women seen consecutively between June 1990 and August 1991 at an obstetrical clinic in Memphis, Tenn, who were enrolled in a visitation program for 2 years after the birth of their first child. PARTICIPANTS: A cohort of 743 women who were primarily black, were pregnant for less than 29 weeks, had no previous live births, and had at least 2 sociodemographic risk factors (unmarried, <12 years of education, or unemployed). INTERVENTION: An average of 7 (range, 0-18) home visits during pregnancy and 26 (range, 0-71) from birth to the child's second birthday. MAIN OUTCOME MEASURES: Rate of subsequent pregnancy, mean interval between first and second birth, and mean number of months of welfare use. RESULTS: Compared with the control group, women who received home visits by nurses had fewer subsequent pregnancies (1.15 vs 1.34; P=.03), fewer closely spaced subsequent pregnancies (0.22 vs 0.32; P=.03), longer intervals between the birth of the first and second child (30.25 vs 26.60 months; P=.004), and fewer months of using Aid to Families with Dependent Children (32.55 vs 36.19; P=.01) and food stamps (41.57 vs 45.04; P=.005). Compared with the effect of the program while the program was in operation, the effect after it ended was essentially equal for Aid to Families with Dependent Children, greater for food stamps, greater for rates of closely spaced subsequent pregnancies, and smaller for rates of subsequent pregnancy overall. CONCLUSIONS: We found enduring effects of a home visitation program on the lives of black women living in an urban setting. While these results were smaller in magnitude than those achieved in a previous trial with white women living in a semirural setting, the direction of the effects was consistent across the 2 studies.

Black or African American↗

PubMed2007

Functionalized hydrogel surfaces for the patterning of multiple biomolecules.

Patterning of multiple proteins and enzymes onto biocompatible surfaces can provide multiple signals to control cell attachment and growth. Acrylamide-based hydrogels were photo-polymerized in the presence of streptavidin-acrylamide, resulting in planar gel surfaces functionalized with the streptavidin protein. This surface was capable of binding biotin-labeled biomolecules. The proteins fibronectin and laminin, the enzyme alkaline phosphatase, and the photo-protein R-phycoerythrin were patterned using soft lithographic techniques. Polydimethylsiloxane stamps were used to transfer biotinylated proteins onto streptavidin-conjugated hydrogel surfaces. Stamped biomolecules were spatially resolved to feature sizes of 10 mum. Fluorescence measurements were used to assess protein transfer and enzyme functionality on modified surfaces. Our results demonstrate that hydrogel surfaces can be patterned with multiple proteins and enzymes, with retention of biological and catalytic activity. These surfaces are biocompatible and provide cues for cell attachment and growth. (c) 2006 Wiley Periodicals, Inc. J Biomed Mater Res 2007.

Animals↗

PubMed2005

Printing biomacromolecules on a bovine serum albumin precursor layer.

Various biomacromolecules including proteins and polysaccharides are printed on a substrate capped with a bovine serum albumin (BSA) precursor layer to create clear co-patterns of these molecules. Characterizations by confocal laser scanning microscopy (CLSM) and atomic force microscopy (AFM) demonstrate the successful production and clear boundaries of the co-patterns. Rinsing the BSA-adsorbed substrate and the biomacromolecules-inked stamp before microcontact printing (microCP) is crucial for the creation of clear and stable co-patterns. The patterns are mainly stabilized by electrostatic interactions and van der Waals forces. Characterizations by ellipsometry, UV-Vis and fluorescence spectroscopy reveal that printing by a flat PDMS stamp yields a denser layered structure of proteins with a higher amount than that of adsorbed proteins. By printing, however, a lower enzymatic catalytic activity for horseradish peroxidase (HRP) or binding capability for avidin (both normalized to amount) is determined. A conformational transition from alpha-helix to beta-sheet of HRP is observed by ATR-IR. By contrast, a BSA precursor layer can effectively improve the functionality of the printed HRP or avidin and preserve the original conformation of the proteins, although the absolute transferred amount of these proteins is decreased.

Animals↗

PubMed2006

Comparing programs for rigid-body multiple structural superposition of proteins.

Different programs and methods were employed to superimpose protein structures, using members of four very different protein families as test subjects, and the results of these efforts were compared. Algorithms based on human identification of key amino acid residues on which to base the superpositions were nearly always more successful than programs that used automated techniques to identify key residues. Among those programs automatically identifying key residues, MASS could not superimpose all members of some families, but was very efficient with other families. MODELLER, MultiProt, and STAMP had varying levels of success. A genetic algorithm program written for this project did not improve superpositions when results from neighbor-joining and pseudostar algorithms were used as its starting cases, but it always improved superpositions obained by MODELLER and STAMP. A program entitled PyMSS is presented that includes three superposition algorithms featuring human interaction.

Amino Acid Sequence↗

PubMed1992

Multiple protein sequence alignment from tertiary structure comparison: assignment of global and residue confidence levels.

An algorithm is presented for the accurate and rapid generation of multiple protein sequence alignments from tertiary structure comparisons. A preliminary multiple sequence alignment is performed using sequence information, which then determines an initial superposition of the structures. A structure comparison algorithm is applied to all pairs of proteins in the superimposed set and a similarity tree calculated. Multiple sequence alignments are then generated by following the tree from the branches to the root. At each branchpoint of the tree, a structure-based sequence alignment and coordinate transformations are output, with the multiple alignment of all structures output at the root. The algorithm encoded in STAMP (STructural Alignment of Multiple Proteins) is shown to give alignments in good agreement with published structural accounts within the dehydrogenase fold domains, globins, and serine proteinases. In order to reduce the need for visual verification, two similarity indices are introduced to determine the quality of each generated structural alignment. Sc quantifies the global structural similarity between pairs or groups of proteins, whereas Pij' provides a normalized measure of the confidence in the alignment of each residue. STAMP alignments have the quality of each alignment characterized by Sc and Pij' values and thus provide a reproducible resource for studies of residue conservation within structural motifs.

Algorithms↗

PubMed1976

[Some mechanical properties of the pig aorta (author's transl)].

In a machine for tensile tests the ultimate strength, stress and strain of the aorta ascendens of 47 female pigs (German landrace) at three different regions of the vessel wall were determined. The animals were fattened under identical conditions in a progeny testing station and slaughtered at the same living weight (102 +/- 2 kg). Weak regressions of body weight and age on parameters measured were nevertheless accounted for. The average elastin content of the aorta ascendens diminished with advancing age but was not significantly correlated to biomechanical properties of the three single samples. Maximum load, stress and modulus of vessel wall segment I (stamped out nearer to the heart) was greater than in sample II. These circumferential segments however exhibited highly stronger biomechanical resistance than sample III, which was stamped longitudinally. In samples I and II there was a negative correlation between thickness of the wall and ultimate tensile stress (N/mm2). Freezing of the material prior to test at -18degrees C for one week resulted in a significant rise in ultimate tensile strength and stress. The meaning of these findings for comparative angiology is discussed.

Age Factors↗

PubMed1994

Effects of morphine and naloxone on behaviour in the hot plate test: an ethopharmacological study in the rat.

The objectives of this study were: i) to analyse the effects of morphine and naloxone on the rat's behaviour in the hot plate test using an ethological approach, and ii) to compare the effectiveness of repeated versus single test paradigms. Animals received either morphine (0, 3, 6 or 9 mg/kg SC) or naloxone (0, 0.01, 0.1 or 1 mg/kg SC). For repeated hot plate measures, rats were tested before and 60, 120, 180 and 240 min following morphine treatment, as well as 30, 60, 90 and 120 min after naloxone injection. For the single test schedule, rats were tested only once 60 min after morphine or 30 min after naloxone administration, or at 60, 120, 180, 240 and 300 min after 9 mg/kg morphine treatment. Behaviour was videotaped and analysed by an ethogram and ethological techniques. A cluster analysis revealed that the most frequently displayed patterns could be categorised into exploratory sniffing reactions (walk-sniff, immobile-sniff) and noxious-evoked elements, including primary (paw-licking, stamping), escape (jumping, leaning posture) and independent (hindleg-withdrawal) patterns. During repeated tests, morphine treatment induced: i) a maximum hypoalgesic effect 60 min post-injection (noxious-evoked patterns were significantly reduced), and ii) an unexpected "thermal hyperreactivity rebound effect" after 120 min (paw-licking and hindleg-withdrawal were enhanced), although changes in hindpaw-licking are more indicative of a hyperalgesic rebound effect. Most changes were quite similar during the single test schedule at 60 and 120 min after morphine injection. With regard to naloxone treatment, jumping latency was significantly decreased during the repeated test schedule, but not on single exposure to the plate. Other elements were facilitated, however, in the single test (stamping, leaning posture, hindleg-withdrawal). The results indicated that both repeated and single tests paradigms are of value for testing the effects of morphine and naloxone on rats. However, under our conditions the single test paradigm gave a better picture of the overall effects of the drug. Learning as well as habituation and sensitization may mask certain effects during repeated tests. In conclusion, an ethological analysis of the rat's behaviour in the hot plate test following administration of morphine and naloxone has been validated in this study.

Animals↗

PubMed1979

A microtechnique for the rapid analysis of macromolecule synthesis in minicultures of mammalian cells.

A new micromethod, called the Stanzen technique, is described for the rapid determination of DNA and protein content as well as the incorporation rates of radioactively labeled precursors into macromolecules in cells growing in replica minicultures on plastic petri dishes. The procedure yielded reproducible results assaying only minimal cell numbers per sample and was applied for studying both primary or early passaged cell cultures (mouse epidermal cells and fibroblasts) and a malignantly transformed epidermal cell line. In four well defined circular areas (called Stanzen) marked on the bottom of tissue-culture plastic petri dishes (by heated stamps), 0.2 to 4 x 10(5) cells per area were plated and grown as four individual cultures in one dish. Both treatment and labeling with radioactive precursors of these Stanzen cultures were performed as with normal petri dishes. After fixation and extraction of the cultures, the singular Stanzen areas (with the cells fixed onto them) were sawed out and transferred into vials for liquid-scintillation counting or determination of DNA and protein. The obtained values of specific activity corresponded well whether the samples compared were derived from the minicultures of the same dish or from several dishes. By modifications of the known colorimetric methods for DNA and protein determination, the sensitivity of these procedures was improved down to values of 1 microgram DNA or 5 micrograms protein per individual culture. These micromodifications yielded the same values as the standard methods whether applied to cell suspensions or to cell cultures. Finally, cell proliferation was not influenced by the growth conditions in the small Stanzen areas and proceeded as in normal dishes or larger areas similarly stamped on the bottom of petri dishes. Since this method proved valuable for biochemical studies using primary cultures of mouse epidermal cells (saving cell material by a factor of 10, test substances and time), it might also be advantageous for other purposes as well where the availability of cells or test substances are limiting factors for large test series.

Animals↗

PubMed2005

Rapid fabrication and chemical patterning of polymer microstructures and their applications as a platform for cell cultures.

Much of the current knowledge regarding biological processes has been obtained through in-vitro studies in bulk aqueous solutions or in conventional Petri-dishes, with neither methodology accurately duplicating the actual in-vivo biological processes. Recently, a number of innovative approaches have attempted to address these shortcomings by providing substrates with controlled features. In particular, tunable surface chemistries and topographical micro and nanostructures have been used as model systems to study the complex biological processes. We herein report a versatile and rapid fabrication method to produce a variety of microstructured polymer substrates with precise control and tailoring of their surface chemistries. A poly(dimethylsiloxane) (PDMS) substrate, produced by replication over a master mold with specific microstructures, is modified by a fluoro siloxane derivative to enhance its anti-adhesion characteristics and used as a secondary replication mold. A curable material, deposited by spin coating on various substrates, is stamped with the secondary mold and crosslinked. The removal of the secondary mold produces a microstructured surface with the same topographical features as the initial master mold. The facile chemical patterning of the microstructured substrates is demonstrated through the use of microcontact printing methods and these materials are tested as a platform to guide cell attachment, growth and proliferation. The master mold and flexible fluorinated PDMS stamps can be used in a repeated manner without any degradation of the anti-adhesion characteristics opening the way to the development of high-throughput fabrication methods that can yield reliable and inexpensive microstructured and chemically patterned substrates.

Animals↗

PubMed1993

Structure of the rat's behaviour in the hot plate test.

The objectives of the study were: (i) to elaborate an ethogram of the rat's behaviour during the hot plate test, (ii) to analyse the rat's responses to the nociceptive heat stimulus by using ethological techniques, and (iii) to discern the suitable pattern or patterns to evaluate the rat's reaction to the noxious stimulus. Male Wistar rats (n = 30) were employed, and one hot plate test was carried out for each rat. The platform (Socrel DS37) was maintained at 55.0 +/- 0.5 degrees C, and the exposure time was 45 s. Behaviour was videotaped and analysed by an ethogram and a software package. Every pattern parameter (frequency, duration and latency for the first occurrence) was quantified. Cluster and Markovian analyses were also employed, whereby a dendrogram and a Markov chain were obtained, respectively. The ethogram was composed of twelve patterns: immobile-sniff, walk-sniff, rearing, forepaw-licking, hindpaw-licking, face-washing, body-cleaning, stamping, hindleg-withdrawal, leaning posture, jumping off, and freezing. Cluster analysis indicated that the most frequent patterns could be classified into three categories: sniffing responses (immobile-sniff, walk-sniff), primary noxious-evoked patterns (forepaw-licking, hindpaw-licking, stamping), and escape noxious-evoked responses (leaning posture, jumping off). Hindleg-withdrawal would be an independent noxious-evoked pattern, and freezing an alerting behaviour. The ethological information could be useful to better elucidate effects on rats' behaviour of drug treatment or other phenomena during the hot plate test.

Animals↗

PubMed2024

Identification of RBP binding sites using RNA deaminases.

RNA-binding proteins (RBPs) are critical regulators of gene expression and RNA processing. Identification of their binding sites has important implications for their physiological and disease-related functions. Crosslinking and immunoprecipitation, followed by sequencing (CLIP-seq) and its derivatives, are the most commonly used methods to identify RBP binding sites, but are laborious and require a large amount of starting material. Recent advancements harnessing RNA deaminases in fusion to any RBP of interest, allow for the profiling of RBP binding sites from low-input samples in simpler procedures. Among these efforts, we developed STAMP (Surveying Targets by APOBEC-Mediated Profiling), which efficiently detects RBP-RNA interactions. This chapter describes the detailed protocol for the STAMP method, including plasmid construction, delivery and sorting, library preparation and bioinformatic data analysis.

RNA-Binding Proteins↗

PubMed2004

A randomized trial of amifostine and carmustine-containing chemotherapy to assess lung-protective effects.

We conducted a randomized, double blind, placebo-controlled multi-institutional trial to assess the ability of amifostine to protect patients against acute lung injury associated with cyclophosphamide/cisplatin/carmustine (BCNU) (STAMP I), a BCNU-containing high dose chemotherapy regimen used with hematopoietic cell transplantation. Amifostine was administered in a dose of 740 mg/m(2) for 2 doses preceding administration of BCNU, the presumed pulmonary-toxic component of the regimen. The trial was stopped after 79 patients were randomized and a planned interim analysis demonstrated that it was unlikely that pulmonary cytoprotection would be detected with further accrual. We conclude that amifostine, used in the dose and schedule we tested, does not reduce the incidence of acute lung injury produced by STAMP I. Further, we suggest that amifostine use with BCNU in other contexts and with clinically achievable doses is unlikely to protect the lung from BCNU-associated acute injury.

Adult↗