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Comparison of waterless hand antisepsis agents at short application times: raising the flag of concern.

OBJECTIVE: Although alcohol-based hand rinses and gels have recommended application times of 30 to 60 seconds, healthcare workers usually take much less time for hand hygiene. We compared the efficacies of four alcohol-based hand rubs produced in Europe (hand rinses A, B, and C and one gel formulation) with the efficacy of the European Norm 1500 (EN 1500) reference waterless hand antisepsis agent (60% 2-propanol) at short application times. DESIGN: Comparative crossover study. SETTING: Infection Control Program laboratory of a large tertiary-care teaching hospital. PARTICIPANTS: Twelve healthy volunteers. INTERVENTION: Measurement of residual bacterial counts and log reduction factors following inoculation of fingertips with Staphylococcus aurens American Type Culture Collection (ATCC) 6538, Pseudomonas aeruginosa ATCC 15442, and a clinical isolate of Enterococcus faecalis. RESULTS: All hand rinses satisfied EN 1500 standards following a single application for 15 and 30 seconds, but reduction factors for the gel formulation were significantly lower for all tested organisms (all P < .025). CONCLUSIONS: Under stringent conditions similar to clinical practice, all three hand rinses proved to be more efficacious than the marketed alcohol-based gel in reducing bacterial counts on hands. Further studies are necessary to determine the in vivo efficacy of alcohol-based gels and whether they are as efficacious as alcohol-based rinses in reducing the transmission of nosocomial infections.

Anti-Infective Agents, Local↗

Performance evaluation of microPET: a high-resolution lutetium oxyorthosilicate PET scanner for animal imaging.

UNLABELLED: A new dedicated PET scanner, microPET, was designed and developed at the University of California, Los Angeles, for imaging small laboratory animals. The goal was to provide a compact system with superior spatial resolution at a fraction of the cost of a clinical PET scanner. METHODS: The system uses fiberoptic readout of individually cut lutetium oxyorthosilicate (LSO) crystals to achieve high spatial resolution. Each microPET detector consists of an 8 x 8 array of 2 x 2 x 10-mm LSO scintillation crystals that are coupled to a 64-channel photomultiplier tube by optical fibers. The tomograph consists of 30 detectors in a continuous ring with a 17.2-cm diameter and fields of view (FOVs) of 11.25 cm in the transaxial direction and 1.8 cm in the axial direction. The system has eight crystal rings and no interplane septa. It operates exclusively in the three-dimensional mode and has an electronically controlled bed that is capable of wobbling with a radius of 300 microm. We describe the performance of the tomograph in terms of its spatial, energy and timing resolution, as well as its sensitivity and counting-rate performance. We also illustrate its overall imaging performance with phantom and animal studies that demonstrate the potential applications of this device to biomedical research. RESULTS: Images reconstructed with three-dimensional filtered backprojection show a spatial resolution of 1.8 mm at the center of the FOV (CFOV), which remains <2.5 mm for the central 5 cm of the transaxial FOV. The resulting volumetric resolution of the system is <8 microL. The absolute system sensitivity measured with a 0.74 MBq (20 microCi) 68Ge point source at the CFOV is 5.62 Hz/kBq. The maximum noise equivalent counting rate obtained with a 6.4-cm diameter cylinder spanning the central 56% of the FOV is 10 kcps, whereas the scatter fraction is 37% at the CFOV for an energy window of 250-650 keV and the same diameter cylinder. CONCLUSION: This is the first PET scanner to use the new scintillator LSO and uses a novel detector design to achieve high volumetric spatial resolution. The combination of imaging characteristics of this prototype system (resolution, sensitivity, counting-rate performance and scatter fraction) opens up new possibilities in the study of animal models with PET.

Animals↗

Changes in some factors of the innate immunity and serum zinc and iron concentrations in pigs following intravenous administration of Escherichia coli lipopolysaccharide.

The changes in some factors of the innate immunity (phagocytosis, complement, lysozyme); haematological parameters-leukocytes, erythrocytes, differential white blood cell counts, haemoglobin, haematocrit and the serum concentrations of the microelements zinc and iron in six 2- to 3-months-old female piglets after the intravenous administration of lipopolysaccharide from Escherichia coli 0111:B4 were determined. It was found out that 1 h after the administration of lipopolysaccharide at the dosage rate of 10 microg/kg body weight resulted in a decrease in the phagocytic parameters, i.e. the phagocytic number and the index of phagocytic activity, which was followed by an increase in their values between post treatment hours 2 and 4. The leukocyte counts had decreased by hour 2 after the injection, but thereafter increased, and at post treatment hour 72, a leukocytosis was observed. The differential white blood cell counts were characterized by a shift to the left between hours 2 and 4 and a statistically significant increase in lymphocyte counts at hour 48 of the experiment. The serum zinc concentrations were increased an hour after the lipopolysaccharide application; after which their average values were lower. The haemolytic activities (CH50) of the classical and the alternative pathways of complement activation decreased. The haemolytic activity (CH50) for the classical pathway began to increase at hour 48 following the treatment. Significant changes were not observed in lysozyme activity, serum iron concentrations or the related haematological parameters (erythrocytes and haemoglobin).

Animals↗

Automatic cell culture quantitation with TRAKCELL: application to cell toxicology and differentiation.

An automated system, TRAKCELL, was developed for the quantitation of cells in culture. It enabled cell counting, classification according to morphological cell characteristics and measurement of cell proliferation and differentiation. The system was tested on the toxic effect of ascorbic acid on rat brain catecholaminergic neurons in primary culture. In parallel, the effects of nerve growth factor, dexamethasone and forskolin on cell differentiation were studied using rat pheochromocytoma PC12 cells. The results show that the system permits rapid and reproducible measurements of cell density and of the morphological changes observed following various drug treatments.

Animals↗

Liquid scintillation radioassay in disposable microcentrifuge tubes: radioimmune precipitates and other applications.

A simple, economical radioassay system employing disposable polypropylene microcentrifuge tubes was developed. Plastic adapters permitted automatic operation in liquid scintillation spectrometers. Counting efficiencies of (3)H, (14)C, (32)P, and (125)I in liquid scintillation cocktails and of (32)P by Cerenkov radiation (at lower efficiency in absence of added scintillator) were comparable to those in standard vials. Multipurpose use of the microtubes made the system versatile and expedient, e.g., collection of precipitates and radioassay in the same container. Collection of radioimmune precipitates was aided by a carrier inorganic precipitate, Mg(2)P(2)O(7).

Animals↗

Assay of neutralizing antibody against variola virus by the degree of focus reduction on HeLa cell cultures and its application to revaccination with smallpox vaccines of various potencies.

A method for assaying neutralizing antibody against variola virus was established by focus counting on HeLa cell cultures. The ND(50) titre, i.e., the serum dilution endpoint to give a 50% reduction in the number of foci, was determined with excellent reproducibility.Groups of students 19-20 years of age were revaccinated by the multiple pressure method with serial 10-fold dilutions of a smallpox vaccine and their neutralizing antibody response was assayed by the focus counting assay system and was related to the local skin reactions on the seventh day after inoculation and to the potency of the vaccine administered. There was a significant rise in the antibody level even after inoculation with a vaccine whose potency was as low as 1.3 x 10(5) pock-forming units/ml. In general, the rise in the log antibody level was proportional to the diameter of the reddening, but a significant rise was found among individuals who had no detectable skin reaction. The skin reaction was greater among individuals with a lower initial antibody level when the vaccine administered had a potency lower than 1.3 x 10(6) pock-forming units/ml.

Adult↗

[Determination of the cell count in sheep's milk].

A prospective study was done to examine the extent to which the 'regulation concerning the method of examining milk using the Coulter counter' was also applicable to sheep's milk. This was found to be the case, provided the milk is previously mixed with a Vortex mixer for 0.5 minutes. In view of the studies done by others, the threshold value was reduced to 4.5 mu. The levels recorded by this method were readily reproducible. Any statement regarding the relationship between the cell counts recorded and the cells actually present in the milk cannot be based on these studies.

Animals↗

Exploiting the effect of dietary supplementation of small ruminants on resilience and resistance against gastrointestinal nematodes.

This paper focuses on targeted nutritional supplementation as a means to reduce the requirement for chemotherapeutic control of gastrointestinal nematode infection of small ruminants and considers the limitations to practical application. Supplementary feeding, particularly with additional dietary protein, can assist resilience to infection during times when metabolic resources are being directed towards dealing with the pathophysiological effects of infection and away from production of meat, milk and fibre. Substantial experimental evidence from studies of both sheep and goats supports this hypothesis particularly in relation to young lambs and kids after weaning and in ewes around parturition. In addition, nutritional supplementation frequently increases resistance to infection, as indicated by decreased faecal worm egg counts and worm burdens. As a result, supplementation has the potential to reduce the requirement for anthelmintic treatment. Practical application of this knowledge can, however, be quite complex in many small ruminant production systems. In general, strategic supplementation should target those times when nutrient requirements are greatest and provide those nutrients which are deficient whether protein, energy, minerals or trace elements. Complexity arises when we consider that nutrient requirements will differ between localities for different species and breed of host, at different stages of growth and reproduction, with differing seasonal availability of forage, with different species of nematodes and different levels of established infections and exposure to infective stages. As a starting point, the provision of nutrients to optimize rumen function and animal performance in the particular production system should assist in maintaining resilience to nematode infection. Provision of nutrients in excess of this requirement, if economically feasible, may yield further benefits in some situations and reduce the need for alternative control measures for gastrointestinal nematode parasites.

Animal Feed↗

Relationship between CD4 count and CD4% in HIV-infected people.

OBJECTIVE: To describe the relationship between absolute CD4 count and CD4%, and the influence on this of gender, risk group, age, a diagnosis of AIDS, use of zidovudine (ZDV) therapy and PCP prophylaxis. METHODS: 9203 paired serial measurements of CD4 count and CD4% on 1017 initially AIDS-free and ZDV-naive HIV positive patients from a London-based cohort were available for analysis. Multi-level regression procedures were used on log-transformed data to relate values of CD4 count to a given level of CD4%. We estimated the effect of selected covariates on this relationship from the exponent of the covariate coefficient. RESULTS: A strong linear relationship was found between log CD4 and log CD4%, CD4 = e 1.78(CD4%)1.26 or 5.93 (CD4%)1.26 (excluding covariates). Based on this model, a CD4% of 5%, 15%, and 30% corresponded to an estimated CD4 count (95% confidence interval [CI]) of 45 cells/mm3 (17-117 cells/mm3), 182 cells/mm3 (64-499 cells/mm3) and 438 cells/mm3 (132-1395 cells/mm3), respectively. However, after adjustment for selected covariates, the predicted CD4 count for a given CD4% was found to be lower among heterosexuals and injecting drug users as compared with homosexual men by 30% and 17% respectively; following an AIDS diagnosis by 21%; and after initiation of ZDV therapy and PCP prophylaxis by 19% and 10%, respectively. CONCLUSION: This analysis should be useful to clinicians and researchers in relating values of CD4 count to CD4%, although we have demonstrated that this is not a simple relationship. The wide CI observed in the estimated CD4 count particularly at high CD4% values, and the adjustments necessary according to risk group, following an AIDS diagnosis and use of ZDV and PCP therapy limit its application in the clinical setting.

Adolescent↗

A functional role of cholinergic innervation to neurons in the cat visual cortex.

1. Effects of microionophoretic application of acetylcholine (ACh) and its antagonists on neuronal responses to visual stimuli and to electrical stimulation of the lateral geniculate nucleus were studied in the cat striate cortex. 2. Responses elicited visually and electrically were facilitated by ACh in 74% of the cells tested, whereas the responses were suppressed in 16%. These ACh effects were blocked by a muscarinic antagonist, atropine, but not by a nicotinic antagonist, hexamethonium, indicating that the ACh effects are mediated through muscarinic receptors. A single application of atropine suppressed visual responses of cells facilitated by ACh, whereas it enhanced those of cells inhibited by ACh, suggesting that endogenous ACh may tonically modulate visual responsivity of cortical neurons. 3. In most cells with the facilitatory ACh effect, responses with single spikes to the electrical stimulation became more consistent, often with double spikes, during the ACh application. The suppressive effects of ACh were noted most often in cells with a longer response latency to electrical stimulation of lateral geniculate nucleus. 4. In most of the facilitated cells the spontaneous activity remained null or very low during ACh application, in spite of marked enhancement of visual responses, suggesting that ACh may improve the signal-to-noise ratio (S/N) of cortical neuron activity. To confirm this suggestion, we calculated a S/S + N index by counting the total number of spikes in the responses (S) and that in peristimulus time histogram (S + N) and found that it was improved during the ACh application in about a half of the cells, whereas it became worse in about one-fifth. 5. In most of the facilitated cells, ACh enhanced visual responses not only to optimal but also to nonoptimal stimuli, resulting in no improvement or even worsening of the orientation selectivity. This was also the case in the selectivity of direction of stimulus movement. 6. The laminar location of the facilitated cells was biased toward layers V and VI of the cortex, although they also made up the majority in layers II + III and about half the tested cells in layers IVab and IVc. 7. In the light of recent understanding of cortical circuitry, these results suggest that the cholinergic innervation to cortical neurons may play a role in improvement of the S/N ratio of information processing in the striate cortex and in facilitation of sending processed informations to other visual centers.

Acetylcholine↗

A successful herd mastitis control scheme in Natal.

Somatic cell counting combined with bacterial identification was successfully used as a method of reducing dairy-herd mastitis problems in Nata. Application of the recommended control measures resulted in an average increase in daily production of up to 2,8 l of milk per cow per day, improvement in quality and reduction in the prevalence of mastitis.

Animals↗

Idiopathic thrombocytopenic purpura in pregnancy and neonatal period.

In pregnancy and neonatal period both mother and child are endangered by bleeding complications due to maternal idiopathic thrombocytopenic purpura. Obstetrical and perinatal management therefore must aim at increasing maternal and fetal platelet count. In our paper six patients in nine pregnancies are reported. Two of them (five pregnancies) were treated with corticosteroids, four of the patients were successfully treated with i.v. immunoglobulins (IgG). Longterm steroid application and splenectomy during pregnancy may be hazardous for mother and fetus. IgG i.v. administration in contrast offers a new and safe way to control maternal and fetal platelet counts during pregnancy, delivery and the neonatal period.

Adult↗

Bacterial penetration and proliferation in root canal dentinal tubules after applying dentin adhesives in vitro.

Endodontic treatment is aimed at eliminating infection and preventing bacterial regrowth in the root canal and dentinal tubules. In the present study the ability of two dentin adhesives to prevent bacterial penetration and subsequent proliferation in dentinal tubules was evaluated. Cylindrical root specimens prepared from freshly extracted bovine teeth were used in an in vitro model of dentinal tubule infection. After removal of the smear layer the intracanal dentinal tubules of the specimens were acid-etched and treated with either Gluma or EBS. Untreated specimens served as controls. Specimens were infected with Enterococcus faecalis and incubated in Brain Heart Infusion for 21 days. Powder dentin samples obtained from within the canal lumina, using ISO 025 to 033 burs, were examined for the presence of vital bacteria by inoculating on agar plates and counting colony-forming units. A significant difference was found between the experimental groups and the untreated group. After application with Gluma specimens showed the least viable bacteria in dentinal tubules. Data suggested that dentin adhesives reduced bacterial invasion into dentin and therefore have a potential role in endodontic treatment.

Acid Etching, Dental↗

Ocular penetration of topical Delta9-tetrahydrocannabinol from rabbit corneal or cul-de-sac application site.

PURPOSE: To determine the relative effects of low-volume drop placement either on the corneal surface or in the lower cul-de-sac on Delta9-tetrahydrocannabinol (THC) penetration into ocular fluids and tissues. METHODS: Pigmented rabbits received 10 microl drops in one eye at either application site. Care was taken to ensure that the drop remained at the site of drug placement. At different intervals after drop administration containing ( 3)H-Delta9-tetrahydrocannabinol in combination with either alpha-cyclodextrin or hydroxypropylmethylcellulose as excipients in different vehicles, eye fluids and tissues were harvested, digested, sampled and counted. RESULTS: Greater quantities of THC were found in the corneal epithelium, stroma/endothelium and aqueous humor of treated eyes after corneal drug application relative to values obtained after cul-de-sac drug application. Cul-de-sac applications gave higher values for inferior bulbar conjunctiva compared with corneal application. These results occurred for each vehicle despite a quantitative difference in drug penetration between the two vehicles. No THC was found in untreated contralateral eyes. CONCLUSIONS: Application of a lipophilic drug to the rabbit corneal surface exposed in the interpalpebral space produced higher tissue and fluid drug concentrations in the anterior segment of the eye compared with drug application into the lower cul-de-sac. This observation has relevance to pre-clinical drug toxicity testing as well as drop placement in patients for the treatment of ocular diseases.

Administration, Topical↗

Human hyperimmune globulin protects against the cytotoxic action of staphylococcal alpha-toxin in vitro and in vivo.

Alpha-toxin, the major cytolysin of Staphylococcus aureus, preferentially attacks human platelets and cultured monocytes, thereby promoting coagulation and the release of interleukin-1 and tumor necrosis factor. Titers of naturally occurring antibodies in human blood are not high enough to substantially inhibit these pathological reactions. In the present study, F(ab')2 fragment preparations from hyperimmune globulin obtained from immunized volunteers were tested for their capacity to inhibit the cytotoxic action of alpha-toxin in vitro and in vivo. These antibody preparations exhibited neutralizing anti-alpha-toxin titers of 80 to 120 IU/ml, whereas titers in commercial immunoglobulin preparations were 1 to 4 IU/ml. In vitro, the presence of 2 to 4 mg of hyperimmune globulin per ml protected human platelets against the action of 1 to 2 micrograms of alpha-toxin per ml. Similarly, these antibodies fully protected human monocytes against the ATP-depleting and cytokine-liberating effects of 0.1 to 1 microgram of alpha-toxin per ml. Intravenous application of 0.5 mg (85 to 120 micrograms/kg of body weight) of alpha-toxin in cynomolgus monkeys elicited acute pathophysiological reactions which were heralded by a selective drop in blood platelet counts. Toxin doses of 1 to 2 mg (170 to 425 micrograms/kg) had a rapid lethal effect, the animals presenting with signs of cardiovascular collapse and pulmonary edema. Prior intravenous application of 4 ml of hyperimmune globulins per kg inhibited the systemic toxic and lethal effects of 1 mg (200 micrograms/kg) of alpha-toxin. In contrast, normal human immunoglobulins exhibited no substantial protective efficacy in vitro and only marginal effects in vivo. It is concluded that high-titered anti-alpha-toxin antibodies effectively protect against the cytotoxic actions of alpha-toxin.

Animals↗

In situ production of a synthetic barrier dressing for burn wounds in rats.

We describe the in situ production of a burn wound dressing applied to eschar that completely isolates burned tissue from contamination with Pseudomonas aeruginosa. Anesthetized, adult rats were subjected to a scald burn over 10% of their body surfaces. One-half hour later a test dressing presumed to be a barrier to bacterial contamination of the wound was applied to the burned surface. Tannic acid, vaseline, ethyl linoleate, collodion, and polyhydroxethylmethacrylate (PHEMA) were evaluated. Each agent was applied directly to the burned surface. A solid film of the PHEMA was produced on the eschar by addition of solvent and a powdered form of the polymer. The surface of each synthetic dressing was contaminated 30 min after application by the addition of 10(8) P. aeruginosa. Also, a control set of rats was burned and their eschars were directly contaminated without application of the test dressing. Seven days later the contaminated muscle under the burned area in 10 control rats had P. aeruginosa counts of 10(7) to 10(8) per g of muscle (wet weight). Of the materials tested, only PHEMA consistently acted as an effective barrier dressing, reducing bacteria in the muscle to undetectable levels in 11 of 14 tests. It was also possible to treat contaminated eschar through this synthetic dressing by topical application of antibiotics to the barrier surface. The results suggest a novel clinical approach in which a barrier dressing could be used to isolate a burn eschar from environmental and subject contamination until the wound site is ready for grafting.

Acrylic Resins↗

The use of vinca alkaloids in adult patients with refractory chronic idiopathic thrombocytopenia.

The aim of the study was to evaluate the application of vincristin or vinblastin in patients with chronic idiopathic thrombocytopenic purpura (ITP), resistant to corticosteroids or with partial contraindication to their application. Twenty-two patients were treated with vincristin or vinblastin in doses of 2 and 10 mg, respectively. Eight of these patients were additionally administered prednisone in an oral dose of 0.5 mg/kg body mass (bm). Two-hour intravenous infusions of drugs were made once a week, at least three times. In every patient, the platelet count was evaluated before and after the three infusions. A rise of the platelet count of at least 100 x 10(9)/l was assumed to signify improvement. Statistically significant improvement (P <0.01) was obtained in nine (41%) patients (including five (35%) patients treated with vinca alkaloids only and in four (50%) patients treated with vincristin and corticosteroids). On the average, 8 weeks after the termination of the treatment there was a 40% drop in the platelet. Patients with a shorter duration of the disease and without detectable platelet antibodies responded well to the treatment more frequently. Minor complications were observed in five patients (23%), notably in the form of paresthesia. Leukopenia was not present. Vinca alkaloids could find their application in clinical situations requiring short-term increase of the platelet count in chronically ill patients with ITP, resistant to corticosteroids or with counterindication to their application.

Adrenal Cortex Hormones↗

Combined high pressure and temperature induced lethal and sublethal injury of Lactococcus lactis--application of multivariate statistical analysis.

It was the aim of this work to determine the combined effects of pressure, temperature, and co-solutes on Lactococcus lactis, and to detect correlations between culture-dependent and culture-independent methods for assessment of cellular viability and sublethal injury. Therefore, the pressure induced inactivation of L. lactis MG 1363 was investigated in buffer and in buffer with 1.5 M sucrose or 4 M NaCl at a pressure range of 0.1 to 500 MPa and a temperature range of 5 to 50 degrees C. The inactivation was characterised by viable cell counts, stress resistant cell counts, membrane integrity, metabolic activity, and the activity of the multi-drug-resistance transport enzyme LmrP. L. lactis was most resistant to pressure application at 20-30 degrees C. Sucrose protected towards inactivation at any temperature, NaCl provided protection at high temperatures only. By using Principal Component Analysis, correlations were detected between viable cell counts and metabolic activity as well as stress resistant cell counts and LmrP activity. In conclusion, the pressure-inactivation of L. lactis is strongly temperature dependent, baroprotection by sucrose occurs at any temperature but the baroprotective effects of NaCl is temperature dependent. Further on, a combination of two experimental methods fully describe lethal and sublethal injury of pressure treated cells. These simplification of data acquisition and model development facilitates the establishment of pressure processes in food technology.

Bacterial Proteins↗