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PubMedDate not supplied

Umbilical vein white blood cell count as a marker of acidemia in term neonates.

OBJECTIVE: White blood cells are mobilized under both hypoxic and infectious conditions. Intrauterine hypoxia is linked to increased risk of cerebral palsy and is potentiated by the presence of infection. We hypothesized that umbilical vein white blood cell elevation in term neonates is associated with intrauterine acidemia. METHODS: We prospectively evaluated all liveborn neonates delivered at our institution for a 6-month period. Umbilical arterial blood was analyzed for pH and blood gas and venous blood for hematologic indices. Medical records of cases greater than or equal to 37 weeks' gestation were reviewed for correlative data. Student's t-test was used to determine difference of means and Chi-square test for goodness of fit. Pearson coefficients of correlation were applied where appropriate. RESULTS: A total of 1,948 liveborn, term neonates were delivered during the study period; 1,561 cases had white blood cell analysis and arterial blood gas data available. Acidemic cases had higher white blood cell (15.0 vs. 12.4 cells x 10(3)/mm3, P < 0.001), lymphocyte (4.43 vs. 3.59 cells X 10(3)/mm3, P < 0.0001), and neutrophil counts (9.08 vs. 7.71 cells x 10(3)/mm3, P < 0.01). As umbilical artery pH decreased, white blood cells became more prevalent. Likewise, as base deficit deepened, white blood cell counts increased. CONCLUSIONS: This study demonstrates an association between deepening acidemia and increasing white blood cell, lymphocyte, and neutrophil counts. Although statistically different, mean white blood cell counts for acidemic and nonacidemic cases are fairly close, limiting the clinical applicability in determining whether pathology is present in an individual case.

Acidosis↗
PubMed1993

Recombinant human interleukin-8 restores function in neutrophils from patients with myelodysplastic syndromes without stimulating myeloid progenitor cells.

Prognosis in myelodysplastic syndrome (MDS) is not only correlated closely with blast cell count in bone marrow and chromosomal abnormalities but also correlated with decreased leucocyte count and function leading to acquisition of lethal infections. Recently, clinical trials in MDS have focused on the application of haemopoietic growth factors such as G-CSF or GM-CSF, which have proven to increase neutrophil count and function. However, these cytokines carry the risk of stimulating the malignant clone, particularly in patients with increased blast cell count. Therefore, investigation of cytokines which are able to stimulate neutrophil function without the potential risk of stimulating haemopoietic progenitor cells may be relevant for MDS. As the stimulatory effect of interleukin-8 on neutrophil function is well known, we investigated whether recombinant human IL-8 is also able to improve the function of neutrophils gained from patients with MDS. Using three different techniques--the E. coli killing assay (8 patients), the production of reactive oxygen as determined by cytochrome c reduction (7 patients) and chemiluminescence (8 patients)--a significant stimulation of neutrophil function at a concentration of 10 nm IL-8 was found in all test systems. No correlation with FAB classification was evident. On the other hand, IL-8 only mildly stimulated growth of myeloid progenitor cells in bone marrow culture of healthy individuals and MDS patients. This minimal stimulation was blocked by a neutralizing antibody directed against GM-CSF, suggesting an indirect effect of IL-8 via secondary GM-CSF release. Thus, IL-8 is able in vitro to repair the functional abnormalities of neutrophils from patients with MDS but has only a marginal influence on myeloid progenitor cells.

Blood Bactericidal Activity↗
PubMed2006

Evaluation of ALOA plating medium for its suitability to recover high pressure-injured Listeria monocytogenes from ground chicken meat.

AIMS: To evaluate a chromogenic plating medium for the isolation of sublethally injured cells of Listeria monocytogenes from processed foods. METHODS AND RESULTS: The inactivation of L. monocytogenes at pressures up to 400 MPa and 12 degrees C in ground chicken meat was employed to examine the recovery of high-pressure injured cells. Before and after different repair incubation periods at 30 degrees C in a nonselective broth, samples were plated onto a selective and differential agar [Agar Listeria according to Ottaviani and Agosti (ALOA)] and in the same medium supplemented with 4% sodium chloride (ALOA-S), and incubated at 37 degrees C. Sublethally injured cells were able to grow when directly plated onto the ALOA medium, without a previous repair incubation period. However, only uninjured cells grew on the ALOA-S medium. CONCLUSIONS: Sublethally injured cells of L. monocytogenes can be quantified by subtracting counts on ALOA-S medium from counts on ALOA medium. SIGNIFICANCE AND IMPACT OF THE STUDY: Possible applications include direct enumeration on ALOA of stressed cells of L. monocytogenes in foods with more than 100 colony forming units per gram.

Animals↗
PubMed2002

Application of flow cytometric techniques to veterinary clinical hematology.

Flow cytometry has emerged as a major new technology in veterinary clinical laboratories. Flow cytometers in current use include stand-alone instruments and cytometers incorporated into hematology analyzers. Flow cytometers offer rapid and quantitative analysis of a variety of cell types based on cell size, molecular complexity, and antigenic composition. Therefore, flow cytometry complements and extends knowledge that can be obtained by light microscopy. Stand-alone instruments are very flexible, however, this flexibility opens the instrument to obtaining invalid or misleading results. The recent development of monoclonal antibodies specific for epitopes on blood cells of food and companion animals has greatly expanded the spectrum of tests with potential clinical application. Tests that appear to have the greatest potential for routine application include reticulocyte and reticulated platelet enumeration, detection of erythrocyte-bound immunoglobulin, immunophenotyping of leukemias and lymphomas, and bone marrow differential cell counting. This report will briefly review the technical aspects of flow cytometry and then focus on techniques with present or potential application to the veterinary clinical laboratory.

Animals↗
PubMed1993

Automated counting of nuclei in series of conventional liver sections to differentiate hypertrophy and hyperplasia.

A method based on image analysis and applicable in pharmacology and toxicology is described that has been designed to meet the statistical requirements for the determination of liver hypertrophy or hyperplasia. An algorithm has been developed to detect and count fluorescent nuclei in Feulgen-stained liver sections (12,500 to 25,000 nuclei or 170 to 350 fields of observation per section within 2-4 hr) by means of fully automatic image analysis with the Leitz Texture Analysis System (TAS) (up to 23 sections in series on the microscope stage). The applicability of this method to conventionally derived, formaldehyde-fixed, tissue sections has been tested on 3-microns sections cut from archived paraplast blocks of already diagnosed materials. Alterations in the liver induced by eight different compounds administered perorally for 28 days to rats have been studied. The morphometric results were found to be specific for a given compound and revealed reproducible and statistically significant dose dependencies of hypertrophy or hyperplasia, or both. Increased frequencies of mitotic figures observed by eye correlated reasonably well with the morphometrically determined hyperplasia, but many hyperplastic livers revealed no mitotic figures. By contrast, the histological diagnoses of an increased degree of hypertrophy were only poorly correlated with the morphometric determinations and, in many instances, were made in livers showing no decrease in the frequency of nuclei. On the other hand, the results from electron microscopy agreed well with the corresponding morphometric analyses: in cases with predominant hypertrophy the proliferation of the smooth endoplasmic reticulum was distinct or striking, or the peroxisomes were altered and increased in number, whereas no obvious ultrastructural changes were seen in cases of morphometrically exclusive hyperplasia.

Animals↗
PubMed1999

Clinical trial of transdermal testosterone and oral levonorgestrel for male contraception.

Approaches to hormonal male contraception are predominantly based on injectable testosterone (T) application. As most users would prefer an injection-independent modality, this study was designed to develop a self-applicable hormonal male contraceptive regimen by combining transdermal T with an oral gestagen. Eleven healthy men (23-40 yr old) were treated with oral levonorgestrel and transdermal T for 24 weeks. T was applied daily as a transdermal patch to be worn on the trunk. Levonorgestrel was taken orally at a dose of 250 microg daily up to week 12, followed by 500 microg to week 24 in those volunteers who had not become azoospermic by that time. Within 24 weeks, 2 of 11 volunteers had become azoospermic, and 3 of 11 showed sperm concentrations below 3 million/mL. The sperm concentrations of the remaining volunteers declined, but failed to reach the limit considered compatible with contraception by WHO. Treatment resulted in suppression of LH, FSH, and sex hormone-binding globulin, whereby the volunteers with lower sperm concentrations showed more pronounced suppression than the others. Mean T concentrations remained within the lower limit of normal and on occasions were below this level. There were no complaints of hypoandrogenism. Although mean levels of low density lipoprotein cholesterol, apolipoprotein B, as well as basal and postprandial insulin increased, high density lipoprotein cholesterol and apolipoprotein A-I decreased during the treatment phase. Changes in lipid parameters were normalized within 3 weeks after cessation of medication. Although only 5 of 11 volunteers reached the target sperm counts (<3 million/mL), the study shows that a self-applicable hormonal male contraceptive could be developed.

Administration, Cutaneous↗
PubMed1999

The effect of intranasal and inhaled corticosteroids in healthy volunteers on the number of circulating lymphocytes and lymphocyte subsets.

BACKGROUND: There has been an increasing interest in the potential systemic effects of inhaled corticosteroids. METHODS: The effect of locally inhaled corticosteroids in the nose and lung on blood lymphocytes was measured in two studies. In the first study, budesonide (BUD) (200 and 800 microg), fluticasone propionate (FP) (200 and 800 microg), and placebo were administered in the nose, and BUD (1600 microg) and FP (1500 microg) were inhaled into the lungs in a blinded, randomized fashion by 12 healthy volunteers. Blood samples were taken before and 4 h after the administration of the drug, and total lymphocyte count and different subpopulations were determined. In the second study, 15 healthy volunteers were randomized to BUD (1600 microg), FP (1600 microg), or placebo inhaled into the lungs. Blood samples were taken before and 4, 8, 24, 48, and 148 h (=7 days) after inhalation of the medication. RESULTS: Neither the nasal applications nor the inhalation of FP (1500 microg/1600 microg) showed significant differences in total lymphocyte count or different subpopulations between baseline and 4 h after the administration. In both studies, a significant reduction was found in the total lymphocyte count, B cells, T cells, and the CD4+ and the CD8+ fractions 4 h after application of BUD 1600 microg. CONCLUSIONS: Nasal application of BUD or FP in doses up to 800 microg do not induce lymphopenia. BUD 1600 microg inhalation in the lung reduces lymphocytes and their subfractions. Further studies have to be done to determine whether the results obtained in this study in healthy volunteers will also be found in patients with diseased mucosa and whether there is any correlation with adverse effects such as growth inhibition or osteoporosis.

Administration, Inhalation↗
PubMed1980

Uses of the UICC samples.

The UICC samples are now used in a wider variety of applications than was originally envisaged. They have been employed in inhalation, ingestion, inoculation and in vitro experiments. Other applications are in identification of asbestos type in aerosols and tissues; standardization of conventional fibre counting; development and calibration of techniques and instruments for fibre assessment; and studies on influence of fibre size and shape on inhalation and retention. The paper outlines applications and gives literature references.

Air Pollutants, Occupational↗
PubMed1998

The antral follicle count predicts the outcome of pregnancy in a controlled ovarian hyperstimulation/intrauterine insemination program.

PURPOSE: Our purpose was to test whether age-related changes in antral follicle counts can predict the pregnancy outcome in the early follicular phase of a controlled ovarian hyperstimulation/intrauterine insemination (COH/IUI) program. METHODS: A selected group of 107 women (36 healthy women requesting child sex preselection, 52 women with unexplained infertility, and 19 with minimal endometriosis) who underwent controlled ovarian hyperstimulation with clomiphene citrate (CC) plus human menopausal gonadotrophin (hMG) and subsequent intrauterine insemination were enrolled in the study. Transvaginal ultrasonography (7.0 MHz) was used to determine the total number of antral follicles (2-8 mm) in the right and left ovaries. The association among the antral follicle count, age, dominant follicle, and estradiol (E2) level on the day of human chorionic gonadotropin (hCG) was analyzed. The association of the pregnancy rate and OHSS with the antral follicle count, dominant follicle count, and age was also examined. RESULTS: The total antral follicle number decreased with age (P < 0.0001). Dominant follicle number increased with total antral follicle number in women who received CC plus hMG/ IUI(P < 0.0001). The pregnant group had a higher number of antral follicle and dominant follicles in comparison with the nonpregnant group (P < 0.01 and P < 0.02, respectively). The E2 level on the day of hCG injection increased positively with the total number of antral follicles (P < 0.0001) and the total number of dominant follicles (P < 0.0001). In women aged younger than 35 years, the pregnancy rate and dominant follicle number rose as the number of antral follicles increased (P < 0.03 and P < 0.0001, respectively). The pregnancy rate was low (2/39) in women aged older than 35 years regardless of the number of antral follicles (P < 0.05) and the extent of hMG administration (P < 0.02). Women aged older than 35 also produced fewer dominant follicles (P < 0.001). No pregnancy was achieved in a patient with an antral follicle number of less than five (17 cases). CONCLUSIONS: Age-related changes in antral follicle count significantly predicted the dominant follicle count and the pregnancy outcome. In women with antral follicle counts of less than five or who are older than 35 years, the application of COH/IUI may not be indicated.

Age Factors↗
PubMed2004

Evaluation of the anthelmintic activity of Khaya senegalensis extract against gastrointestinal nematodes of sheep: in vitro and in vivo studies.

The anthelmintic effect of Khaya senegalensis is described. In vitro and in vivo studies were conducted to determine possible direct anthelmintic effects of ethanolic and aqueous extracts of K. senegalensis towards different ovine gastrointestinal nematode. A larval development assay was used to investigate in vitro, the effect of aqueous and ethanolic extracts towards larvae of strongyles. The LC50 values of the effects of both the aqueous and ethanolic extracts were calculated. Another study was conducted in vivo to evaluate the therapeutic efficacy of the extracts administered orally at a dose rate of 125, 250 and 500 mg/kg of sheep harbouring naturally acquired infection of gastrointestinal nematodes. The presence of K. senegalensis extracts in the cultures decreased the viability of larvae. The LC50 of the aqueous extract (0.69 mg/ml) is not significantly different (P >0.05, t-test) from the ethanolic extract (0.51 mg/ml). The activity of the extract is concentration dependent in vivo. Sheep drenched with 500 mg/kg K. senegalensis ethanolic extract had a mean faecal egg count (FEC) reduction of 88.82%. The extract of K. senegalensis could find application in anthelmintic therapy in veterinary practice.

Animals↗
PubMed1987

Long-term follow-up of men with male pattern baldness treated with topical minoxidil.

Forty-one men with male pattern baldness completed 132 study weeks (2 years 9 months) with topical minoxidil and had follow-up 1-inch target-area vertex scalp hair counts. Initially these men were treated with either twice-daily 2% topical minoxidil for 12 months or 3% topical minoxidil for 8 to 12 months (one third of the subjects received placebo for the first 4 months). After 12 months all subjects continued to apply 3% topical minoxidil twice daily for 1 more year, after which they were randomized to once- versus twice-daily topical minoxidil for an additional 9 months. Those subjects who changed to once-daily application of topical minoxidil at 2 years had a mean change from baseline nonvellus hair count at 1 year of 291.2 (range of hairs four to 553) and at 2 years 9 months of 235 (two to 592 hairs). Those subjects who continued with twice-daily application of topical minoxidil throughout the study had a mean change from baseline nonvellus hair count at 1 year of 323 (15 to 589 hairs) and 335 (13 to 808 hairs) at 2 years 9 months with maintenance topical minoxidil. There were subjects on both maintenance schedules of topical minoxidil who lost some of the nonvellus hair they had initially gained with topical minoxidil; however, there was a greater mean loss in those patients following the once-daily versus twice-daily topical minoxidil regimen (p = 0.05). No subject lost nonvellus target hair as compared with baseline.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Topical↗
PubMed1992

A review of recent advances in stereology for quantifying neural structure.

The science of stereology has undergone a revolution over the past decade with the introduction of design-based (assumption- or model-free) methods which are highly efficient and generally unbiased. No other morphometric approach currently offers these twin benefits. Stereology is ideal for extrapolating 3-D structural quantities (real volumes, surface areas, lengths and numbers) from simple counts made on 2-D slice images. The images may take various forms (e.g. physical or optical sections, MRI slices, CT scans) but they must be sampled so as to be random in orientation and/or position if valid estimates are to be made. All the recent developments in stereology are applicable to problems in neuromorphometry. This review provides an account of major developments and the state of the art, emphasizes the importance of properly randomized sampling and identifies some applications to neural structure at different levels of organization. These include the counting and sizing of synapses, neurites, cells and whole brains.

Animals↗
PubMed1997

Effects of four analgesic treatments on the behavioural and cortisol responses of 3-week-old lambs to tail docking.

The behavioural and cortisol responses of groups of seven or eight lambs were used to determine which of three methods of tail docking (rubber ring, Burdizzo and rubber ring combined, or heated docking iron) produced the least signs of pain in the first 3 h after use and which of four analgesic treatments (1.0 ml bupivacaine subcutaneously, 0.5 ml bupivacaine epidurally, a topical cold analgesic spray or diclofenac 1.5 mg kg-1) was most effective in reducing these signs. Amputation with a heated docking iron produced levels of behaviour and cortisol responses which did not differ markedly from those of handled controls. The rubber ring method produced the greatest increase in all parameters (total active behaviour 110 +/- 91 counts; 51 +/- 23 min spent in abnormal postures; peak cortisol 93 +/- 51 nmol l-1). Subcutaneous bupivacaine, administered immediately prior to application of the ring, appeared to be the analgesic treatment most effective at reducing these responses (23 +/- 15 counts; 24 +/- 22 min.; 44 +/- 20 nmol l-1).

Administration, Topical↗
PubMed2006

Automated tool for the detection of cell nuclei in digital microscopic images: application to retinal images.

PURPOSE: To develop an automated tool that provides reliable, consistent, and accurate results for counting cell nuclei in tissue sections. METHODS: We propose a novel method based on an image processing algorithm to analyze large sets of digital micrographs. The nucleus detector design is based on a Laplacian of Gaussian filter. We use the leave-one-out cross validation method for estimating the generalization error, which is then used to choose the model and parameters of the proposed nucleus detector with both fluorescent and dye stained images. We also evaluate the performance of a nucleus detector by comparing the results with manual counts. RESULTS: When our nucleus detector is applied to previously unanalyzed images of feline retina, it correctly counts nuclei within the outer nuclear layer (ONL) with an average error of 3.67% ranging from 0 to 6.07%, and nuclei within the inner nuclear layer (INL) with an average error of 8.55% ranging from 0 to 13.76%. Our approach accurately identifies the location of cell bodies. Even though we have a relatively large error in the INL due to the large intra-observer variation, both manual counting and nucleus detector result in the same conclusion. This is the first time that cell death in the INL in response to retinal detachment is analyzed quantitatively. We also test the proposed tool with various images and show that it is applicable to a wide range of image types with nuclei varying in size and staining intensity. CONCLUSIONS: The proposed method is simple and reliable. It also has widespread applicability to a variety of sample preparation and imaging methods. Our approach will be immediately useful in quantifying cell number in large sets of digital micrographs and from high-throughput imaging. The tool is available as a plug-in for Image J.

Algorithms↗
PubMedDate not supplied

A new coincidence model for single particle counters, Part II: Advances and applications.

Accuracy, acceptance limits and methods for U.S.P. (788) contaminating particle assays published in the XXII Revision are refined in U.S.P. XXIII. In both Revisions, although different numerical values and methods are employed, particle contamination limits remain constants for all S.V.I. container volumes. The effect of this quality standard is high particle concentration acceptance limits in the smallest S.V.I. container sizes. The effect of these high concentrations is to introduce both undercount errors and false counts into U.S.P. (788) SVI contaminating particle assays. There is general agreement that the count of high concentrations of particles by a single particle light extinction counter result in an increase of the average size of the distribution of particles reported and a decrease in their total number. The error mechanism is termed "signal coincidence." Understanding and control of both these problems is unified with the introduction of the count efficiency parameter. Part I of this paper makes available two core concepts with which evaluation and control of coincidence error in single particle counters can be accurately quantified. These two core concepts are the "Particle Triggered Poisson Model," a new more accurate statistical model of the particle counting process and a concentration measure that includes the effect of particle size on the counting capability of a detector. Use of these concepts make it possible to evaluate particle detector count efficiency capability from experimental data of the coincidence effect. This is an application paper. It combines the theory in the Part I paper with the replicability of particle counters into a simple test protocol. The test results can be used to calculate a contour of particle size and count within which both undercount errors and the introduction of false counts into U.S.P. (788) particle assays are controlled. From the data analyzed it can be seen that any single particle size test cannot effectively evaluate detector performance. The use of the theory and methodology described can help realize the intent of the U.S.P. (788) SVI particle contamination assay.

Calibration↗