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Effect of dietary n-6-to-n-3 fatty acid ratio on complete blood and total white blood cell counts, and T-cell subpopulations in aged dogs.

OBJECTIVE: To determine effect of diets with variable n-6-to-n-3 fatty acid (FA) ratio on CD4+ and CD8+ T-lymphocyte subpopulations, and on results of routine laboratory analyses (CBC and total WBC count, serum biochemical analyses, and urinalysis). ANIMALS: 20 healthy, aged (9.5 to 11.5 years old) female Beagles. PROCEDURE: Dogs were fed 1 of 3 diets that contained 6% fat by weight but differed in amounts of n-6 and n-3 FA. For 11 weeks, 6 dogs were fed a low concentration of n-3 FA (ratio, 31:1), 7 were fed a medium concentration (5.4:1), and 7 were fed a high concentration (1.4:1). Preprandial blood and urine samples were collected before beginning the study and at 8 weeks for evaluation of laboratory variables. Before and at 3, 6, and 8 weeks during the study, blood was drawn for total WBC and lymphocyte counts and for characterization of T-cell subpopulations. At 8 and 10 weeks, dogs were vaccinated with keyhole limpet hemocyanin suspension. Blood was drawn 4 days after each vaccination, and lymphocytes were isolated for flow cytometry. Effects of diet and vaccination on each variable were determined. RESULTS: After vaccination, total lymphocyte count increased and CD4+ T lymphocyte count and the CD4(+)-to-CD8+ ratio decreased in dogs consuming the diet with n-6-to-n-3 FA ratio of 1.4:1. CONCLUSION: Feeding a diet with n-6-to-n-3 FA ratio of 1.4:1 had significant effects on CD4+ T lymphocytes in healthy, aged Beagles after vaccination.

Aging↗

Impact of visceral involvements and blood cell count abnormalities on survival in adult T-cell leukemia/lymphoma (ATLL).

Multiple visceral involvements and various blood cell count abnormalities are frequently manifested in adult T-cell leukemia/lymphoma (ATLL) at diagnosis. We evaluated the effects of four visceral involvement (bone marrow (BM), skin, liver, spleen) and six blood cell count abnormalities (anemia, neutrophilia, thrombocytopenia, monocytosis, eosinophilia, basophilia) on the overall survival of 168 ATLL patients. In the aggressive type, BM involvement, skin involvement and monocytosis were significantly poor prognostic factors. Furthermore, concomitant involvement of BM and additional visceral organs worsened the prognosis. These data support that multiple organ involvements represent a poor prognostic factor for ATLL and provide clinical significance for BM examinations.

Adult↗

Somatic cell counts in bovine milk: relationships to production and clinical episodes of mastitis.

The relationships between somatic cell counts, milk production and episodes of clinical mastitis were evaluated using data collected between 1979 and 1981 in 32 southern Ontario Holstein herds. Somatic cell counts were logarithmically transformed and the distribution of the resulting counts is presented. The seasonal pattern in cell counts was evaluated using a formal statistical procedure. Counts were lowest in the winter and spring and highest in the early fall but the differences amongst monthly geometric mean cell counts were small. Assuming a linear relationship between log somatic cell counts and test day milk production it was found that a unit increase in the log count resulted in a loss of 1.44 kg of milk. Regression analyses within specific log cell count ranges indicated that the previous estimate may underestimate losses at low cell counts and overestimate losses at higher cell counts. The relationships between cell counts and episodes of mild or acute clinical mastitis were evaluated by comparing counts preceding and following the clinical episodes to comparable counts in matched control cows. Mild cases of mastitis were preceded by higher cell counts than were found in control cows but the same phenomenon was not observed in acute cases of mastitis. Both mild and acute cases were followed by higher cell counts than were found in control cows.

Animals↗

Correlation between circulating stromal cell-derived factor 1 levels and CD4+ cell count in human immunodeficiency virus type 1-infected individuals.

Stromal cell-derived factor 1 (SDF-1) is the natural ligand that recognizes CXCR4, which also serves as a coreceptor for some strains of HIV-1. In this study, we explored SDF-1 blood levels among HIV-1-infected individuals exhibiting a wide range of CD4+ cell counts. Plasma or serum concentrations of SDF-1 protein were measured by ELISA in samples from 31 HIV-1-seronegative individuals and 79 HIV-1-infected subjects. Although SDF-1 protein levels were stable for months among seronegative individuals (mean intrasubject variation, 17%), the absolute values varied widely (0.28 to 106.5 ng/ml; mean, 25.6 ng/ml). In HIV-1-infected subjects, there was a direct correlation between SDF-1 level and CD4+ cell count. Subjects with fewer than 50 CD4+ cells per cubic microliter of blood had significantly lower mean SDF-1 levels (+/-SD) than did either HIV-1-infected subjects with higher CD4+ cell counts or uninfected controls: CD4+ cell count <50, mean SDF-1 level of 10.7+/-33.7, 50 < CD4+ cell count <200, mean SDF-1 level of 12.9+/-19.0, 200 < CD4+ cell count <500, mean SDF-1 level of 19.3+/-36.8; CD4+ cell count >500, mean SDF-1 level of 18.5+/-25.2; uninfected control mean SDF-1 level, 25.6+/-34.7. No significant change in SDF-1 level was detected after administration of antiretroviral therapy in nine subjects with advanced disease (mean intrasubject variation, 43%). Analysis of SDF-1 mRNA expression in lymph nodes from HIV-1-infected subjects at different disease stages revealed that the medullary cords contained stromal cells that express SDF-1 mRNA. This preliminary analysis suggests a possible link between lower SDF-1 levels and disease progression.

Adult↗

CMVR diagnoses and progression of CD4 cell counts and HIV viral load measurements in HIV patients on HAART.

AIM: To assess the impact of highly active antiretroviral therapy (HAART) on the prevalence and progression of CMV retinitis (CMVR) among AIDS patients with baseline CD4 cell counts <100 cells x 10(6)/l. METHODS: A longitudinal cohort study of 1292 patients. CD4 cell counts and HIV viral load measurements were obtained before commencing therapy, at 3 months, 1 year, 2 years, and at last follow up. The CMVR prevalence rate was measured for the subgroup with baseline CD4 cell counts <100 cells x 10(6)/l. CMVR adverse event (AE) rates per 100 person days at risk were calculated for the subgroup with CMVR and baseline CD4 cell counts <100 cells x 10(6)/l. RESULTS: 1292 patients were started on HAART. 8% of patients had CD4 counts <50 cells x 10(6)/l and 40% had detectable HIV viral load at last follow up. The prevalence of CMVR for the subgroup with baseline CD4 <100 cells x 10(6)/l was 10%. For those with baseline CD4 <100 cells x 10(6)/l, the mean CMVR AE rate was greatest during the first 6 months of follow up after HAART commencement (p <0.003). The mean AE rate per 100 person days at risk was 0.36 (95% CI 0.167 to 0.551) before starting HAART, and 0.14 (95% CI 0.085 to 0.199) after starting HAART (p = 0.03). CONCLUSIONS: HAART significantly prolongs the disease-free intervals in patients with pre-existing disease but recurrences persist within the first 6 months of starting therapy. AE were absent beyond 18 months of follow up in all patients including those with persistently low CD4 counts and detectable HIV viral load indicating clinical immunorestoration. New methods for monitoring the response to therapy are needed to identify those at risk.

Adult↗

A precise group size in Dictyostelium is generated by a cell-counting factor modulating cell-cell adhesion.

A remarkable aspect of Dictyostelium development is that cells form evenly sized groups of approximately 2 x 10(4) cells. A secreted 450 kDa protein complex called counting factor (CF) regulates the number of cells per group. We find that CF regulates group size by repressing cell-cell adhesion. In both experiments and computer simulations, high levels of CF (and thus low adhesion) result in aggregation streams breaking up into small groups, while no CF (and thus high adhesion) results in no stream breakup and large groups. These results suggest that in Dictyostelium and possibly other systems a secreted factor regulating cell-cell adhesion can regulate the size of a group of cells.

Animals↗

Changes of activated macrophages and apoptotic cell count in the organs of rats during experimental sepsis.

OBJECTIVE: Leukocyte migration plays an important role in inflammation. The aim of our study was to detect the activated macrophage count and to define the apoptotic cell count in the rat's liver and lungs during different stages of sepsis and to clarify whether the activity of macrophages is associated with the apoptotic cell count in the course of the septic process. MATERIAL AND METHODS: Experimental sepsis was induced by inoculating Wistar rats intraperitoneally with E. coli cells. The liver and lung tissue was obtained 2, 6, 24, 48 and 120 hours after inoculation, and blood smears to detect the leukocyte volume were prepared at the same time. Macrophage activity was studied by immunohistochemical staining, using the ABC method. Apoptotic cell count was detected with the "In Situ Cell Death Detection Kit". RESULTS: Decrease in the total count of leukocyte and lymphocyte percentage and a rise in the percentage of neutrophils in the blood were evidences of a strong inflammation process in an organism. The count of activated macrophages in the liver was high, showing the maximum level at the end of the 2(nd) h after inoculation, after which it began to fall. Apoptotic cell count rose after a decrease in macrophage activity. In the lungs both changes - a decrease in activated macrophage level and a rise in the apoptotic cell count took place later on. CONCLUSION: Our investigations indicate that a decrease in the activated macrophage level is connected with an increased rise in the apoptotic cell count in both organs depending on the stage of the disease and occurs in the liver earlier than in the lungs but the process of cell apoptosis was more intensive in the lungs.

Animals↗

Seasonal variation of bulk milk somatic cell counts in UK dairy herds: investigations of the summer rise.

Individual cow somatic cell count (SCC) patterns were explored over a one year period in 33 dairy herds to investigate the reason for a summer rise in bulk milk somatic cell counts (BMSCC). Cow test day somatic cell counts were categorised according to the magnitude of change since the previous test day reading, to examine which categories were responsible for the summer increase. Multilevel models using Markov chain Monte Carlo methods were specified to estimate the number of somatic cells/ml produced by different cell count categories. Stage of lactation and parity were accounted for in the models. There was an increase in the proportion of cows that remained above 200,000 cells/ml for two consecutive recordings in summer and this group of cows were responsible for 70.8% of the increase in somatic cells/ml produced from May to September compared with October to March. There was no evidence that a greater new infection rate (somatic cell counts moving from below 100,000 cells/ml to over 200,000 cells/ml) contributed to the increased summer bulk milk somatic cell counts. There was no indication that a general small increase in all somatic cell counts played an important role in the increased summer somatic cell counts. Markov chain Monte Carlo methods provided a valuable and flexible platform for parameter estimation in reasonably complex multilevel models.

Animals↗

Monitoring udder health and milk quality using somatic cell counts.

In this article the use of somatic cell counts for monitoring udder health and milk quality is discussed. Somatic cell count dynamics at quarter, cow, herd and population level are discussed and illustrated with examples. Quarter and cow somatic cell counts directly represent the inflammatory status of the mammary gland. Herd and population somatic cell count are related to the inflammatory process in individual cows but much more reflect the udder health status of the herd and the quality of the raw milk in the herd and the population. Application of monitoring tools in herd health management are illustrated using a case study. Understanding infection dynamics requires precise longitudinal data. Monitoring tools are required to find the areas of risk in the herd. It is inevitable that more complete udder health programs and monitoring systems are to be developed and implemented. These programs are necessarily dynamic and complex. Implementation of complete udder health programs should be accompanied by research efforts to further fine-tune these complete udder health control and monitoring programs.

Animals↗

Correlation study of two methods for evaluating corneal endothelial damage in vitro: the Janus green photometry technique versus cell counting.

Counting of endothelial cells after vital staining is a time-consuming method which, in addition, requires considerable experience and does not normally allow for the study of the entire endothelial cell surface. However, the Janus green photometry technique (JGPT) is an objective, time- and personnel-saving procedure which also has the advantage of allowing for the evaluation of the whole endothelial cell surface. A comparative simultaneous experimental study of the two methods had not been reported. Freshly excised pig corneas (n = 58) were preserved in MK storage medium for 4, 7, 14 and 20 days at 4 degrees C. All corneas were stained with Janus green and alizarin red. A 7-mm corneal button corresponding to an endothelial surface of 38.46 mm2 was punched out. The percentage of damaged cells was determined with cell counting (CC) and with JGPT, and the results were statistically compared. Two methods for CC were used. Cells were counted either directly under the microscope or in photomicrographs. The overall correlation between the results of CC and JGPT was R = 0.98. However, when endothelial cell damage was higher than 30%, CC became an estimation by approximation, since necrotic areas were confluent, whereas with JGPT the results were usable even when the damage was 100%. Thus it appears that although the two methods can be used interchangeably, JGPT may in some instances be the method of choice.

Animals↗

Genetic and phenotypic relationships between lactation cell counts and milk yield and composition of Holstein cows.

Monthly somatic cell count data were collected between February 1977 and February 1982 for Holstein cows in 928 herds enrolled on the Quebec Dairy Herd Analysis Service. The geometric mean of the log monthly cell counts was calculated for each lactation. Official lactation records for 305-day milk, fat, and protein yields, and fat and protein percents were obtained for same cows. There were 18,189 cows in first lactation representing 257 sires, 13,225 in second lactation representing 206 sires, and 8,683 in third lactation representing 151 sires. Heritabilities of yield traits and protein percent increased across three lactations. Heritability of fat percent was similar in first and third lactations but decreased slightly in second lactations. Heritability of lactation cell count was small, being least in second lactations. Genetic correlations between lactation cell count and yield traits were positive in first lactations, small and negative in second lactations, and small and positive in third lactations. Genetic correlations between lactation cell count and fat and protein contents were small in the three lactations. Phenotypic correlations between lactation cell count and production traits were small in each of the three lactations. Genetic correlations between yield traits in early lactation and lactation cell count in a subsequent lactation were positive. The genetic correlation between protein percent in an early lactation and cell count in a later lactation was large between first and second lactations, decreased between second and third lactation, and small between first and third lactations. Genetic correlations were small and negative for fat percent.

Animals↗

Mononuclear cells in Japanese encephalitis virus infection: changes in cells counts and specific fluorescence.

Progressive reduction in blood cell counts was observed in mice inoculated intracerebrally (ic) with Japanese encephalitis (JE) virus. No changes were observed in the blood cell counts of mice inoculated intraperitoneally (ip). Reduction in cell counts after a transient rise was noticed in lymph nodes of mice inoculated by either route but the cell counts returned to normal in lymph nodes of ip inoculated mice by the 8th day post inoculation (p.i.). JE virus antigen was demonstrated by immunofluorescence in mononuclear cells from the blood, spleen and lymph nodes starting from the 3rd and 4th day p.i. in mice inoculated ic and ip, respectively. The number of fluorescent cells increased as the infection progressed. The number of fluorescent spleen cells uas higher in ip than in ic inoculated mice. Live virus could only occasionally be demonstrated in the cells.

Animals↗

The repeatability of nonbronchoscopic bronchoalveolar lavage differential cell counts.

Airway inflammation in children can be assessed by nonbronchoscopic bronchoalveolar lavage (BAL). Little is known about the repeatability of cell counts in the BAL obtained. Children (n=43) attending for elective surgery were studied. Cell counts were obtained following a nonbronchoscopic lavage. Two samples were obtained with either: 1) the catheter wedged in the same position (n=21) or 2) the catheter reinserted and wedged again (n=22). Slides (n=30) from nonbronchoscopic lavage samples were selected at random and two independent observers counted 500 cells on each slide on two occasions. The repeatability of the lavage sampling and cell counting was assessed for different cell types. The inter- and intra-observer repeatability for the differential cell counting demonstrated that there was good repeatability for all cell types except lymphocytes (interobserver: Lin's concordance coefficient 0.42; repeatability coefficient 0.66). Quantification of eosinophil (%) was highly repeatable using either method (Lin's concordance coefficient 1) 0.99, 2) 0.95; repeatability coefficient 1) 0.58, 2) 1.36). Nonbronchoscopic lavage is a repeatable technique for the quantification of eosinophils. Variation in the sampling method can be reduced by taking two separate samples and averaging the differential cell counts. Furthermore, increasing the number of cells counted should ensure accurate quantification of lymphocytes.

Bronchoalveolar Lavage Fluid↗

HIV infection, antiretroviral therapy, and CD4+ cell count distributions in African populations.

BACKGROUND: The variability in CD4+ cell counts within and among human immunodeficiency virus (HIV)-positive and -negative African populations has not been explained but has important implications for understanding the incidence of HIV-related opportunistic infections, especially tuberculosis, in both individuals and populations. METHODS: In HIV-negative African adults, CD4+ cell counts vary within populations (interquartile ranges [IQRs], 169-603 cells/microL) and among populations (means vary from 699 to 1244 cells/microL), with similarly wide variations in HIV-positive adults. We developed dynamic mathematical models to predict the distribution of CD4+ cell counts in HIV-positive adults using the distribution in HIV-negative adults. RESULTS: Under the assumption that survival is independent of the CD4+ cell count before seroconversion, we fitted the observed distributions in HIV-positive adults. At a CD4+ cell count of 200 cells/microL, the median life expectancy of HIV-positive Zambians (4.0 years) was predicted to be 1.7 times that of HIV-positive South Africans (2.3 years). CONCLUSIONS: The model provides a way to estimate the changing distribution of CD4+ cell counts and, hence, the changing incidence of HIV-related opportunistic infections as the epidemic matures. This could substantially improve the planning of health services, including the need and demand for antiretroviral therapy. Better data are needed to test the model and its assumptions more rigorously and to fully understand the variability in CD4+ cell counts within and among populations.

Adolescent↗

Potential of differential somatic cell counts as indicators of mastitis in quarter milk samples from dairy cows.

Bacteriological status, somatic cell counts and proportions of lymphocytes, granulocytes and monocytes were determined in 1,659 quarter milk samples from 39 dairy cows. Discriminant analysis was performed in order to assess the ability of total and differential somatic cell counts and combinations of total somatic cell count and each of the differential cell counts, to discriminate between infected and pathogen-free quarters, as well as between quarters infected with minor pathogens and quarters infected with major pathogens. Total somatic cell count classified 82.9% of all quarters correctly with respect to bacteriological status. Differential somatic cell count was less effective than total somatic cell count in discriminating between infected and pathogen-free quarters, as well as between quarters infected with minor vs. major pathogens. Combination of total and differential somatic cell counts did not improve the rate of correctly classified quarters. Inclusion of demographic data into the discriminant function increased the number of quarters correctly classified, mainly through an increase in the proportion of correctly classified infected quarters.

Animals↗

[Factors affecting cell counts in milk from individual cows (author's transl)].

The number of somatic cells was determined in samples from approx. 700 cows in 70 herds, altogether 2570 samples. The average cell count was 250 thousands per ml with range from 7 thousands to 5 mill. The data were analysed with respect to effect of age of the cow, stage of lactation, and test day yield. The analyses showed that the cell count was strongly affected by age and yield. The effect of yield is removed by multiplying the recorded cell count by milk yield (cell count in thousands per ml times milk yield in kg congruent to total cell count in mill.). The cell count increases approximately linearly by increasing age of the cow up to 9--10 years, when it is about five times as large as for two-year-old cows. This increase is partly caused by a higher frequency of infection in older cows, but there was a distinct trend in the same direction even for cows which were noninfected when the sample was collected. After elimination of the effect of yield there was a slight drop in the cell count by advancing lactation.

Age Factors↗

Multiple relapses of human cytomegalovirus retinitis during HAART in an AIDS patient with reconstitution of CD4+ T cell count in the absence of HCMV-specific CD4+ T cell response.

BACKGROUND: While in the past human cytomegalovirus (HCMV) represented the major viral opportunistic pathogen in patients with AIDS, incidence of HCMV disease in HIV-infected patients drastically dropped after introduction of highly active antiretroviral therapy (HAART). However, cases of HCMV disease in HIV-infected patients treated with HAART have been reported. OBJECTIVE: A 38-year-old HIV-infected patient developed HCMV retinitis in May 1999 after reaching a nadir of 69 CD4(+) T cells/microl. HAART and anti-HCMV treatments with parenteral ganciclovir (GCV) were started, resulting in HIV viremia suppression, rise in CD4(+) T cell count to >300 cells/microl and recovery from retinitis. Notwithstanding the apparent immune reconstitution, every attempt to discontinue GCV maintenance treatment was followed by a relapse of retinal lesions. Thus, HCMV-specific CD4(+) cellular immune response was investigated. RESULTS: Lymphoproliferation assay and cytokine flow cytometry analysis were performed repeatedly from November 1999 showing absolute lack of HCMV specific CD4(+) T cell response, in the presence of an efficient lymphoprolipherative response against another pathogen (Candida) or a mitogen (Phytohemoagglutinin). CONCLUSION: In some patients, immune reconstitution after HAART may be only partial, since lack of pathogen-specific CD4(+) T cell response may persist even in the case of a significant rise in the absolute CD4(+) T cell count. This case suggests that immunologic assays investigating specific immune response against HCMV in HIV infected patients may be more useful than the CD4(+) T cell count alone in assessing immune function reconstitution after HAART and in deciding interruption of anti-HCMV secondary prophylaxis.

AIDS-Related Opportunistic Infections↗

Efficiently estimated histologic cell counts.

Three different estimates were compared with complete nuclear counts made over 200 fields (20 sets of 10) selected from histologic sections of 18 breast carcinomas. All counting used a 10 x 10 square grid eyepiece graticule at x 400. Two area sampling techniques agreed well with overall counts (59,210 cells counted in all 200 fields; estimates were 58,500 and 59,805 cells), but an estimate based on linear intersection of nuclei yielded a lower estimate (23,382 cells). The most efficient estimate was 10 times the number of carcinoma cell nuclei in a random sample of 10 small graticule squares; summation over 10 fields yielded a mean ratio of estimated to counted total of 98.2% with limits of agreement of 89.9% to 107% for individual-case 10-field estimates. To improve efficiency further a simplified version of this estimate was devised and validated on carcinoma cell nuclei in 200 fields (20 sets of 10) selected from 20 invasive cervical carcinomas. An invariant sampling set was randomized to one of four rotations (0 degree, 90 degrees, 180 degrees, or 270 degrees). A total of 102,799 cells was counted in all 200 fields; the estimate was 101,280 cells. Summation over 10 fields yielded a mean ratio of estimated to counted total of 98.3% with limits of agreement of 89.8% to 108%. This simplified procedure facilitated estimation of cell numbers without detriment to agreement with denumerated counts and could be implemented as a custom-made counting graticule.

Breast Neoplasms↗