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PubMed1985

[Biometric parameters of ovaries in the Tsigai strain of sheep and their changes after administration of cloprostenol].

The weight, length, width and volume of ovaries were determined as well as the number of follicles visible on the surface in relation to the presence of corpus luteum on the ovary in autumn period after Cloprostenol application. The ovaries of 43 sheep of the Tsigai breed were examined at the age of 2--4 years. The animals were divided into five groups. First group (I) was the control (n = 6). In the luteal phase of sexual cycle the animals of groups II to V were applied i. m. 125 micrograms of cloprostenol in Oestrophan inf. Spofa preparation. According to groups II (n = 8), III (n = 10), IV (n = 9), V (n = 10) the animals were killed at intervals of 24, 48, 72 and 120 hours (h) from the preparation application. The removed ovaries were fixed in 10% neutral formalin; after a 48-hour fixation they were weighed and their length and width were measured. Tertiary follicles visible on the surface were counted and measured. In group I we demonstrated the significantly higher weight of ovaries with corpus luteum (CL) as compared with ovaries without CL (P less than 0.001). The weight of ovaries with CL dropped significantly within 48 hours after cloprostenol application as compared with control (P less than 0.001) and the difference in the ovary weight according to CL incidence disappeared almost completely. In comparison with groups III and IV, the weight of ovaries in ewes of group V increased statistically significantly (P less than 0.01) on 5th day from the cloprostenol application. This finding is a result of development of new CL after a passed ovulation. The alterations in length, width and volume of ovaries were not so significant as those in weight. The number of surface follicles was very variable and the changes were not significant. The changes in the average size of follicles larger than 1 mm and of the largest follicles have shown that both parameters achieved significantly higher values in ovaries of the control group with present CL in comparison with ovaries without CL. The reduction of surface follicles after cloprostenol application seems to be connected with a possible operation on contractile structures of the external theca folliculi.

Animals↗
PubMed1984

An electrostatic precipitator for the study of airborne radioactivity.

An system has been developed to measure airborne radioactivity using electrostatic precipitation for collection and alpha-particle spectroscopy for detection. Features include good energy resolution (e.g. 170 and 300 KeV for full-width half maximum and full-width tenth maximum for 7.7-MeV alpha particles using a 7-cm2 area detector; and 52- and 122-KeV, respectively, using 1.2-cm2 area detector) and versatile computer control for collection, counting and data reduction. Aerosols bearing the radioactive atoms are deposited on a foil tape by electrostatic precipitation for a predetermined time after which the foil is moved under a solid-state detector to count the alpha-particle emissions. Activities are determined at the same frequency as samples are collected. Helium gas can be introduced at the detector to reduce energy loss and improve resolution. Although in principle certain aerosol sizes could be difficult to collect, in practice no difficulties were observed for typical environmental conditions, provided sufficiently low air-sampling rates were used. One important application is the measurement of 222Rn daughters. The sensitivity is such that detection of individual daughter concentrations less than 0.1 pCi/l. with only a 10% counting error is possible.

Air Pollution, Radioactive↗
PubMed1999

Adjusting for measurement error to assess health effects of variability in biomarkers. Multicenter AIDS Cohort Study.

Longitudinal studies of health effects often relate individuals' biomarker levels to disease progression. Repeated measurements also provide an opportunity to assess within-individual biomarker variability, and it is reasonable to postulate that this measure might provide additional information about a particular outcome variable. Given the existing precedent for application of adjustment methods to account for measurement error in subject-specific average levels of a covariate, this concept motivates the application of such methods to incorporate variability as well. In this paper, we investigate the nature of the relationship between the decline of CD4 cell count induced by infection with human immunodeficiency virus, and CD4 level and variability prior to infection. We first describe the distribution of repeated CD4 measurements prior to infection using a model that accounts both for random average levels and random subject-specific variance components. Based on this model, we define true unobservable random variables that correspond to prior level and stability. We perform a linear regression analysis, using these latent variables as covariates, by means of a full maximum likelihood approach. We compare the resulting parameter estimates with those based on regressions employing sample-based estimates of pre-infection levels and variances, and empirical Bayes estimates of these quantities. Although the final inferences are similar to those based on the unadjusted analysis, we find that the magnitude of association with prior level decreases, while that with prior stability increases. Stratified analyses indicate that smoking status affects the relationship between prior CD4 level and initial CD4 decline. We point out advantages associated with the maximum likelihood approach in this particular application.

Biomarkers↗
PubMed1999

Comparison of several digital and stereological methods for estimating surface area and volume of cells studied by confocal microscopy.

BACKGROUND: The implementation of different methods for estimating the surface area and volume of cells studied by confocal microscopy was developed. The methods were compared from the point of view of their precision, applicability and efficiency. METHODS: Interactive stereological methods (spatial grid method, fakir method, Cavalieri principle) as well as automatic digital methods (digital Crofton method, voxel counting, triangulation method, iso-intensity contouring method) were considered. The methods were tested on model geometrical solids and on real volume images consisting of a stack of serial sections encompassing entire tobacco BY-2 cells or cell chains. RESULTS: It is shown that many of the studied methods are very precise when applied to cells of simple or moderately complex shapes. The automatic digital methods are fast and precise but their applicability is limited by the necessity to segment automatically the object surface and to find an optimal resolution. This limitation is not present in stereological methods which are applied interactively and thus are more time-consuming. CONCLUSIONS: The presented implementations of the fakir method and the Cavalieri principle enable interactive, unbiased and efficient estimation of the cell surface area and volume. The recommended steps for measuring the surface area and/or volume of objects studied by confocal microscopy are described.

Cell Line↗
PubMed1983

General definition of valence delta-values for molecular connectivity.

The molecular connectivity valence delta-values have been defined in terms of the count of nonhydrogen valence electrons on a valence-state atom as screened from the nucleus by the core electrons. The core is defined as the nonvalence electrons minus 1. This general definition expresses the valence delta-values for second and third quantum level atoms and halogens. Valence delta-values have been derived for higher oxidation states of sulfur and phosphorus. The internal consistency of these delta-values is tested by their ability to closely correlate molar refraction values with 1 chi v. It is found that a second variable, the count of the number of alpha hydrogen atoms, greatly increases the quality of the correlation. Some biological SAR applications reveal the general utility of these findings.

Chemical Phenomena↗
PubMed1992

Electrofusion-induced intracellular Ca2+ flux and its effect on murine oocyte activation.

These experiments were designed to monitor influx of extracellular Ca2+ into the murine ooplasm following a 1.56 kV.cm-1 direct current (DC) electrofusion pulse and subsequently to determine its effect on rate of activation. Pulse media consisted of non-electrolyte (0.3 M mannitol) and electrolyte (phosphate-buffered saline; PBS) media each containing 0.0, 0.05, or 0.9 mM Ca2+ (groups T1-T3 and T4-T6, respectively). Cumulus-free oocytes were incubated in 100 microliters drops of PBS containing 2 microM of the calcium indicator fluo-3/AM for 60 min at 37 degrees C. Fluo-3/AM-loaded oocytes were equilibrated for 7 min in assigned treatment media (T1-T6) prior to application of DC pulse. Change in fluorescent intensity was monitored for 6.5 min after DC pulse by photon counting spectrofluorometry. Fluorometric measurements demonstrate a dramatic rise in intracellular free Ca2+ (Ca2+i) following DC pulse is associated with Ca2+ ion concentration in the pulse medium. Significantly (P less than 0.01) higher Ca2+i levels were observed when 0.9 mM Ca2+ was added to the pulse medium (T3 and T6) compared with pulse medium containing lower Ca2+ ion concentrations (T1, T2, T4, and T5; P greater than 0.05). Differences (P less than 0.01) were observed in peak Ca2+i levels 18 sec after pulse with mean percent change in fluorescence of 5.1%a, 33.9%b, 112.7%c, 1.2%a, 9.3%a, and 99.9%c for T1-T6, respectively (values with different superscripts are significantly different at P less than 0.01). Increased oocyte membrane permeability to Ca2+ ion after DC pulse was observed for a minimum of 5 min after delivery of the 1.56 kV.cm-1 pulse.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1982

Topical and systemic glucocorticoid potencies of budesonide and beclomethasone dipropionate in man.

Topical anti-inflammatory (cutaneous "vasoconstriction") and systemic glucocorticoid (depression of plasma cortisol and changes in differential WBC count) potencies of the two glucocorticoids budesonide and beclomethasone dipropionate (BDP) were compared in human volunteers. After topical application, budesonide was 2-3 times more potent than BDP in inducing "vasoconstriction". After oral administration, on the other hand, budesonide was 2-4 times less potent than BDP in depressing plasma cortisol and changing the total or differential WBC. After inhalation, too, significant differences in favour of budesonide were noted, but the divergence between the drugs was less pronounced. The improved relationship between the topical and systemic glucocorticoid effects of budesonide makes it a promising alternative for aerosol treatment in asthma.

Administration, Oral↗
PubMed2002

[Intracoronary radiation therapy in controlled and open clinical trials with afterloading systems and "hot" balloon catheters. Analysis of 6,692 patients].

The prevention and treatment of a restenosis, which occurs in ca. 30% of the cases following balloon dilatation of coronary stenoses, using intravascular radiation relies on the inhibition of proliferation that is inherent in every radiation therapy. The analysis is based on 6,692 patients assigned to either a control group (1,717 patients) or to radiation therapy (4,975 patients) in 41 studies. A total of 14 placebo-controlled, randomized and 27 open trials have been completed: 22 regarding in-stent restenosis, ten regarding de-novo stenosis (or restenosis without a stent) as strict inclusion criteria, and nine with all types of stenoses. For in-stent restenoses, vessel size as defined for inclusion was between 2.0 mm and 5.5 mm, stenosis length between 10 mm and 80 mm. In all trials with in-stent restenosis, the primary endpoints were reached; the restenosis rate in the longest coronary segment analyzed was between 45% and 100% in the control groups and between 7.7% and 53.5% in the brachytherapy groups. The respective values for the TVR were between 24.1% and 80% in the control groups and between 2.0% and 41.7% in the brachytherapy groups. In the control groups, MACE was between 25.9% and 80%; it was between 2.0% and 41.7% in the brachytherapy groups. Attaining results for de-novo stenoses was problematic due to "geographic miss" apparently playing a larger role in these cases; but when taking this into consideration, good results were also attained. The known limitations due to late stent thromboses (4-15% in older trials) were to the most part eliminated by administration of clopidogrel for 1 year and the limitations due to the "edge effect" by the application of longer radiation sources. With antiproliferative coated stents as treatment for de-novo stenoses, we can count on intracoronary brachytherapy losing significance in this area. But for in-stent restenoses, intracoronary brachytherapy is the only evidence-based interventional form of therapy.

Aged↗
PubMed1981

Radionuclide evaluation of peripheral circulatory dynamics: new clinical application of blood pool scintigraphy for measuring limb venous volume, capacity, and blood flow.

A new application of clinical blood pool imaging is described for measurements of limb venous volume and blood flow. Forearm count rate changes determined using blood pool imaging of in vivo Tc-99m labelled red blood cell (RBC) were compared in the same limb to forearm volume changes measured by Whitney mercury-in-silicone rubber strain gauge in eight normal subjects. Intravascular count rate changes closely correlated with volume changes following sudden (mean r = 0.98) and stepwise (mean r = 0.96) limb venous congestion for assessment of venous compliance, as well as for measurement of limb blood flow up to 20 ml/min/100 ml (r = 0.95). These results indicate the accuracy and utility of the radionuclide technique in the study of peripheral circulatory dynamics in clinical settings. In addition, limb tissue fluid accumultion may also be quantified by simultaneous use of radionuclide-strain gauge plethysmography.

Blood Circulation↗
PubMed1992

Mathematical assessment of the spatial distribution of Langerhans cells in guinea pig epidermis.

Two mathematical indexes, Hopkins-Skellam index (HSI) and Morisita index (MI), were applied to assess the distribution of ATPase-stained epidermal Langerhans cells (ELC) in the guinea pig skin. To our knowledge, this is the first report in which the degree of regularity has been expressed numerically by computation based on theoretical equations. The regularity of ELC in the normal skin was confirmed by the value of HSI (P < 0.0001). In the topical steroid applied skin, the number of ELC decreased significantly but the value of HSI was similar to that of the normal skin; while in the ultraviolet B (UVB) exposed skin, both number of ELC and regularity of ELC distribution decreased significantly. The graph of MI for the UVB exposed skin clearly showed that the distribution of ELC had local clumps. The two indexes, HSI and MI have been quite useful for determining the regularity of ELC. These indexes may have a wide application to other cells.

Animals↗
PubMed2005

Neutrophil activation occurs in the lower-limbs of patients undergoing elective repair of abdominal aortic aneurysm.

OBJECTIVES: AAA repair is associated with a systemic inflammatory response, mediated in part by neutrophils. The aim of this study was to determine where neutrophil activation occurs. METHODS: Blood was sampled from the femoral vein, portal vein and radial artery of 10 patients undergoing elective AAA repair at four time-points [induction of anaesthesia (systemic sample only), pre-aortic clamp application, pre-clamp removal and after 30min of reperfusion]. Whole blood was analysed for the white cell count, neutrophil count, and for neutrophil CD11b expression. RESULTS: The white cell count and neutrophil counts increased after aortic clamp release. Neutrophil expression of CD11b was significantly higher in the femoral vein than the portal vein and systemic circulation during ischaemia [P=0.001 (FV vs. PV), P=0.017 (FV vs. systemic)] and reperfusion [P=0.001 (FV vs. PV), P=0.013 (FV vs. systemic)]. There were no significant differences in neutrophil CD11b expression between the systemic and portal vein samples at any time. CONCLUSIONS: Ischaemia and reperfusion during abdominal aortic aneurysm repair are associated with a global increase in the white cell count and neutrophil count, but with increased neutrophil CD11b expression only in the femoral vein. This suggests the lower-limbs are sensitive to aortic clamp-related reperfusion injury and may fuel the inflammatory response.

Aged↗
PubMed2005

Objective monitoring of nasal airway inflammation in rhinitis.

Allergic rhinitis is an inflammatory nasal disorder in which a range of different cells participates. A variety of approaches has been used to monitor nasal inflammation objectively to investigate disease processes and to evaluate the effect of therapeutic intervention. These approaches include nasal lavage, nasal cytology, and nasal biopsy, together with the more recently established measurement of nasal nitric oxide (NO) concentration. Although all provide information about nasal mucosal inflammation, the extent of information that can be obtained by each approach, the ease of sampling, and the complexity of sample handling differ. Such considerations influence the choice of approach when measurement of nasal inflammation is to be an objective outcome parameter in a clinical trial. In addition, the choice of approach is also determined by the questions or hypotheses that are to be addressed. Nasal lavage is simple and rapid to perform, is well tolerated, and provides a sample that can provide information about luminal cell recruitment, cell activation, and plasma protein extravasation. Nasal cytology involves sampling and recovering mucosal surface cells. It is also easy to perform and is well tolerated in general, although some find that the procedure causes a transient unpleasant sensation. A differential cell count from the sample provides information about relative cell populations. Both nasal lavage and nasal cytology are readily applicable to clinical trials. Nasal cytology sample handling is easier, but nasal lavage offers the advantage of providing considerably greater information from the sample. Nasal biopsy is a considerably more invasive procedure and requires expertise not only in tissue sampling but also in biopsy processing. Therefore, it is applicable only in specialist centers. However, nasal biopsy is the only sampling technique that directly informs about tissue cellular events, although these may be implied, in part from the other sampling approaches. Tissue specimens can be used to evaluate both protein and gene expression. Measurement of nasal NO involves expensive equipment but provides an instantaneous result, unlike the other approaches, all of which require sample processing and analysis. Recommendations for standardization of measurement have been made, and measures are considered in part to reflect allergic inflammation within the nasal mucosa. The limitations of nasal NO are that it reflects only a certain aspect of allergic mucosal inflammation, and that because a proportion of nasally measured NO is derived from the sinuses under normal circumstances, nasal NO is not specific for nasal disease. The high contribution from the sinus mucosa limits the discriminatory ability of nasal NO to reflect nasal tissue-specific alterations. The incorporation of measures of nasal inflammation in clinical trials has distinguished anti-inflammatory therapy from symptomatic therapy and has the potential to provide information about the efficacy of novel therapies for allergic rhinitis.

Biomarkers↗
PubMed2002

Detection of single molecules: solution-phase single-molecule fluorescence correlation spectroscopy as an ultrasensitive, rapid and reliable system for immunological investigation.

More sensitive techniques in molecular and clinical immunology are essential for the development of reproducible profiles. We have developed a novel methodology named solution-phase single-molecule fluorescence correlation spectroscopy (SPSM-FCS) that fulfils these demands. It is based on the quantification of the probability density of single molecule events in solution is necessary. For example, the Brownian motion of the fluorophore rhodamine-green is detected. Counting about 100,000 photon counts per second and per molecule permits the identification of one single compound. In order to study the applicability of SPSM-FCS in immunology, we have detected and identified a larger nonfluorescent substance in a very complex mixture. The 'unknown' molecules studied were the autoantibodies in serum samples directed against the antigen alpha 3 chain of type IV collagen. In both systems, we were able to characterize the probability density of single fluorescent molecules by means of the averaged absolute molecule number without any calibration. The specific molecules exhibited a Poisson distribution in solution in terms of their 'critical' bulk concentration below about 1 nM. This proof of principle indicates how the SPSM-FCS methodology could be used in immunoassays.

Animals↗
PubMed1985

Treatment of burns in children.

Decreases in mortality from major thermal injury over the last 20 years have been due to advances in resuscitation, control of infection, support of the hypermetabolic response, and early closure of the burn wound. Of these advances in burn care, early wound closure has progressed the most in the last five years. The restoration of the protective functions of the skin is of primary importance to the recovery of the burn patient. Biologic dressings (pigskin, amnion, human skin allograft) when applied to fully debrided, relatively uncontaminated wounds have been shown to adhere to the wound surface, reduce the wound colony counts, limit fluid and protein loss, reduce pain, and increase the rate of epithelialization over that obtained with application of topical antimicrobial agents.

Administration, Topical↗
PubMed2000

Effects of combined exposure of micrococcus luteus to nisin and pulsed electric fields.

Death and injury following exposure of Micrococcus luteus to nisin and pulsed electric field (PEF) treatment were investigated in phosphate buffer (pH 6.8, sigma = 4.8 ms/cm at 20 degrees C). Four types of experiment were carried out, a single treatment with nisin (100 IU/ml at 20 degrees C for 2 h), a single PEF treatment, a PEF treatment followed by incubation with nisin (as before) and addition of nisin to the bacterial suspension prior to the PEF treatment. The application of nisin clearly enhanced the lethal effect of PEF treatment. The bactericidal effect of nisin reduced viable counts by 1.4 log10 units. Treatment with PEF (50 pulses at 33 kV/cm) resulted in a reduction of 2.4 log10 units. PEF treatment followed by nisin caused a reduction of 5.2 log10 units in comparison with a 4.9 log10 units reduction obtained with nisin followed by PEF. Injury of surviving cells was investigated using media with different concentrations of salt. Sublethally damaged cells of M. luteus could not be detected by this means, following PEF treatment.

Anti-Bacterial Agents↗
PubMed1998

Synaptic changes in rat maculae in space and medical imaging: the link.

Two different space life sciences missions (SLS-1 and SLS-2) have demonstrated that the synapses of the hair cells of rat vestibular maculae increase significantly in microgravity. The results also indicate that macular synapses are sensitive to stress. These findings argue that vestibular maculae exhibit neuroplasticity to macroenvironmental and microenvironmental changes. This capability should be clinically relevant to rehabilitative training and/or pharmacological treatments for vestibular disease. The results of this ultrastructural research also demonstrated that type I and type II hair cells are integrated into the same neuronal circuitry. The findings were the basis for development of three-dimensional reconstruction software to learn details of macular wiring. This software, produced for scientific research, has now been adapted to reconstruct the face and skull directly from computerized tomography scans. In collaboration with craniofacial reconstructive surgeons at Stanford University Medical Center, an effort is under way to produce a virtual environment workbench for complex craniofacial surgery. When completed, the workbench will help surgeons train for and simulate surgery. The methods are patient specific. This research illustrates the value of basic research in leading to unanticipated medical applications.

Acoustic Maculae↗
PubMed2001

A new model of visceral pain and referred hyperalgesia in the mouse.

The generation of transgenic mice that lack or overexpress genes relevant to pain is becoming increasing common. However, only one visceral pain model, the writhing test, is widely used in mice. Here we describe a novel model, chemical stimulation of the colon, which we have developed in mice. Mice of either sex were injected i.v. with 30 mg/kg Evan's Blue for subsequent determination of plasma extravasation. For behavioural testing, they were placed on a raised grid and 50 microl of saline, mustard oil (0.25-2.5%) or capsaicin (0.03-0.3%) was administered by inserting a fine cannula into the colon via the anus. Visceral pain-related behaviours (licking abdomen, stretching, contractions of abdomen etc) were counted for 20 min. Before intracolonic administration, and 20 min after, the frequency of withdrawal responses to the application of von Frey probes to the abdomen was tested. The colon was removed post-mortem and the Evan's Blue content measured. Mustard oil and capsaicin administration evoked dose-dependent visceral pain behaviours, referred hyperalgesia (significant increase in responses to von Frey hairs) and colon plasma extravasation. The peak behavioural responses were evoked by 0.1% capsaicin and by 1% mustard oil respectively. The nociceptive behavioural responses were dose-dependently reversed by morphine (ED50 = 1.9 +/- 1 mg/kg s.c.). We conclude that this model represents a useful tool both for phenotyping mutant mice and for classical pharmacology since information on visceral pain, referred hyperalgesia and colon inflammation can all obtained from the same animal.

Abdomen↗