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[Day-to-day variation in cell counts in milk (author's transl)].

The article presents results from two series of milk samples where samples from individual cows were collected at each milking over a period of two weeks. Fat content, protein content and cell count were determined for each sample. The cell counts (in thousands per ml) were transformed to logaritmic values and statistically analysed. As much as 75 to 80% of the variation (expressed as proportion of the total sums of squares) was due to differences "among cows" and about 3% due to differences "among days" during the sampling period. Samples from morning milkings had about 20% lower cell counts than samples from afternoon milkings. The standard deviation among samples from two consecutive milkings of the same cow (after removal of systematic differences between days and between morning and afternoon milkings) was computed as about 30% of the mean (i.e. coefficient of variation approximately 0.3). By combining samples from two milkings the random "error" in cell count can be reduced to about 20% of the mean. The samples exhibited a highly significant, but nummerically small, positive within-cow correlation between fat content and cell count, and between protein content and cell count, while the corresponding correlation between milk yield and cell count was slightly negative (non-significant).

Animals

[The cell count in milk of goats].

Weekly milk samples were taken from mammary halves from 30 goats from 8 different herds each with from 2 to 5 experimental animals throughout their lactation periods. The physiological variations in the cell counts of goats milk during lactation were investigated by a projection microscope. Only small variations were found in the average cell counts in milk samples from goats in the same herd, but big variations from one herd to another (Table I and Fig. 3.). The curves of the average cell counts in milk samples of 5 herds showed a rise at the beginning of the season at pasture (Fig. 1 and Fig. 2). Also in the housed periods the cell count showed transitory rises. The herd with the lowest average cell count (220,000/ml) was milked by an ordinary bucket milking system, while the herd with the highest average cell count (1,490,000/ml) was milked by a pipeline milking system. Five herds milked by machine had an average cell count of 720,000/ml and three herds milked by hand 540,000/ml. All the herds in the material had an average cell count of 680,000/ml.

Animals

Protein, casein, and noncasein protein percentages in milk with high somatic cell counts.

More than 1000 milk samples of individual cows with somatic cell counts ranging from 20,000 to 20,000,000 per ml were analyzed by amido black dye binding for total protein, whey protein, and casein. Ten classes of varying somatic cell counts were established. The mean total protein percentage for normal samples (counts less than 500,000/ml) was 3.2% and different from all the other protein class means, ranging from 3.4 to 3.9%. Total protein content increased with somatic cell count. There was no significant difference among any of the ten classes for casein values. Whey protein percentages for normal samples were different from the rest. Whey protein content increased with somatic cell count.

Animals

[Effect of mouse genotype on the hematopoietic stem cell count. II. The number of hematopietic stem cells in BALB/c and CC57BR strain mice differing by the level of endogenous colony formation].

The number of stem hematopoietic cells in the hematopoietic organs of mice of BALB/c and CC57BR strains and (CC57BRXBALB/c)F1 hybrids was studied by the method of exogenous colony-forming units. The assay of migration of stem cells from the bone marrow to the spleen was carried out. It was found that the spleen and the bone marrow of mice of the studied genotypes contain approximately the same relative number of hematopoietic stem cells. The number of stem cells which migrate from the bone marrow to the spleen is greater in the mice of BALB/c strain than in the CC57BR mice.

Animals

Relationship between herpes simplex virus ulceration and CD4+ cell counts in patients with HIV infection.

OBJECTIVE: To establish the incidence of herpes simplex virus (HSV) ulceration in relation to CD4+ cell counts in HIV-infected patients. DESIGN: Swabs were taken from all ulcerated lesions in HIV-infected patients and cultured for HSV. CD4+ cell counts were performed at regular intervals. SETTING: The HIV unit at a London teaching hospital (the Royal Free Hospital, London, UK). PATIENTS: All HIV-infected patients (n = 500) attending the HIV unit. RESULTS: Two hundred and twenty-three swabs were obtained from 118 patients; 83 (37.2%) swabs from 62 (52.5%) patients were positive for HSV. Of 96 swabs taken from patients with CD4+ cell counts < 50 x 10(6)/l, 56 (58.3%) were positive for HSV, compared with 27 of 127 (21.2%) swabs from patients with higher CD4+ cell counts (P < 0.0001). Of patients with CD4+ cell counts < 50 x 10(6)/l, 37 of 47 (78.7%) had positive cultures compared with 25 of 71 (35.2%) of patients with higher counts (P < 0.0001). This trend was observed with swabs from all body sites; sufficient samples were available from oral and perianal lesions to demonstrate statistical significance (P < 0.0001 and P = 0.007, respectively). CONCLUSIONS: These results show a sharp rise in the incidence of HSV with CD4+ cell counts < 50 x 10(6)/l and thus provide important data for the design of studies of anti-HSV prophylaxis. Furthermore, since nearly 60% of all ulcers in patients with such low CD4+ counts are HSV-positive, we suggest appropriate empirical therapy on presentation.

AIDS-Related Opportunistic Infections

Combined blood cell counting and classification with fluorochrome stains and flow instrumentation.

A multiparameter flow cytophotometer was used to count and classify fixed human blood cells fluorochromed with a mixture of ethidium bromide (EB), brilliant sulfaflavine and a blue fluorescent stilbene disulfonic acid derivative (LN). The system measures light scattered by the cells and absorption at 420 nm for all cells. In addition, nuclear EB fluorescence (540 leads to 610 nm) and cytoplasmic fluorescence from LN (366 leads to 470 nm), brilliant sulfaflavine (420 leads to 520 nm) and EB exicted by energy transfer from LN (366 leads to 610 nm) are measured for all nucleated cells. This information is sufficient to perform red and white blood cell counts and to classify leukocytes as lymphocytes, monocytes, basophils, eosinophils or neutrophils. Light scattering and/or nuclear and cytoplasmic fluorescence values may be further analyzed to obtain the ratio of immature to mature neutrophils. Counts produced by the system are in reasonable agreement with those obtained by electronic cells counting and examination of Wright's-stained blood smears; some discrepancies appear to be due to systematic errors in the manual counting method.

Autoanalysis

Bacteriuria in men infected with HIV-1 is related to their immune status (CD4+ cell count).

OBJECTIVE: Reports from the United States that urinary tract infections (UTI) are more common in homosexual than in heterosexual men have not been confirmed in Europe. The occurrence of several UTI in men infected with HIV-1 has been recorded in The Netherlands. We therefore analysed the relationship between the presence of bacteriuria and the immune status (CD4+ cell count) in these HIV-1-infected patients. DESIGN: Urinary cultures were obtained prospectively for 2 years, during the first visit and every 6 months thereafter, when signs and symptoms of UTI occurred and when patients had fever of unknown origin. CD4+ cell counts were measured at the same time. SETTING: The study was performed at the University Hospital, Utrecht, The Netherlands. PATIENTS, PARTICIPANTS: One hundred and thirty HIV-1-infected men attended our hospital. Data from 98 were analysed. Eighty-nine (91%) of these men were either homo- or bisexual. MAIN OUTCOME MEASURES: Positive urinary culture. RESULTS: Group 1 (CD4+ cell count less than 200 x 10(6)/l) consisted of 47 patients; 30% had at least one period of bacteriuria, with 21 episodes. Group 2 (CD4+ cell count 200-500 x 10(6)/l) consisted of 27 patients; 11% had at least one period of bacteriuria, with five episodes. We did not find bacteriuria in the 24 patients in group 3 (CD4+ cell count greater than 500 x 10(6)/l). The rate of bacteriuria per patient-month, 4 (group 1) versus 2 (group 2), differed significantly (P less than 0.001). A significant relationship between CD4+ cell count and bacteriuria was found (P = 0.00003); no relationship, however, was found with anal intercourse, hospitalization, Karnofsky score, follow-up, or age. CONCLUSION: We conclude that men infected with HIV and presenting with a CD4+ cell count less than 200 x 10(6)/l are at increased risk for bacteriuria.

Adult

Use of monitored CD4 cell counts: predictions of the AIDS epidemic in Scotland: CD4 Collaborative Group.

OBJECTIVE: We describe the CD4 database of the Scottish Immunology Laboratories, and its uses and limitations for making short-term predictions of a CD4 cell count less than or equal to 200 x 10(6)/l (CD4(200)) and of adult AIDS cases in Scotland. DESIGN: The date of the earlier of two consecutive samples (typically 3 months apart) both with CD4 cell counts less than or equal to 200 x 10(6)/l was taken to define when a patient had passed the CD4(200) threshold (referred to as a CD4(200) case). The CD4 database comprises HIV-1-seropositive adults in the four main risk groups [homosexual/bisexual (1), injecting drug users (IDU; 2), heterosexual contact (3), and undetermined (9)] from Scotland's three principal areas of population (Lothian, Tayside and Strathclyde) who have had a CD4 cell count of less than or equal to 500 x 10(6)/l. SETTING: Three hospitals in Scotland, the Communicable Diseases (Scotland Unit) and the Medical Research Council Biostatistics Unit, Cambridge, UK. PATIENTS, PARTICIPANTS: The CD4 database at 31 December 1990 listed 813 patients (of whom 52% were IDU): 390 were CD4(200)/AIDS cases (of whom 44% were IDU) and 192 were AIDS cases (of whom 32% were IDU). RESULTS: Individuals in risk groups 1, 2 and 3 were nearly equally represented among newly diagnosed HIV-1 infections in 1990. However, among patients with moderate immunodeficiency, IDU accounted for 50% of the total number. Co-incidence of first CD4 cell count with CD4(200) diagnosis was recorded for only 28% of IDU, but in over 50% of cases for each of the other exposure groups (57%). There was a highly significant decrease of around 80 x 10(6)/l per calendar-year-of-referral in first CD4 cell counts for patients on the CD4 database; and decreases of around 40 x 10(6)/lper decade of age at referral. Since 1988, median time from CD4(200) to AIDS diagnosis in Scotland has been approximately 2 years. Back-projection was applied to annual CD4(200)/AIDS diagnoses before 31 December 1990 and to AIDS diagnoses. From AIDS diagnoses, the central epidemic scenario underestimated past HIV-1-antibody-positive reports (up to the end of 1985). More dramatic underestimation was occasioned by back-projection from CD4(200)/AIDS diagnoses [319 inferred HIV infections compared with 445 HIV-1-antibody-positive reports to Communicable Diseases (Scotland) Unit]. CONCLUSIONS: First CD4 cell counts should complement new HIV-1 diagnoses. Past referrals for immunological monitoring were not uniform between risk groups in Scotland. Underascertainment of CD4(200) cases is a problem when CD4(200) cases are used as a basis for back-projection. More information concerning the incubation distribution from HIV seroconversion to CD4(200) diagnosis is required. It is likely that there are twice as many CD4(200)/AIDS as there are diagnosed cases of AIDS.

Acquired Immunodeficiency Syndrome

The association between the fat cell count of amniotic fluid and tests of fetal lung maturity.

The fat cell count in amniotic fluid samples from 100 patients was compared with the lecithin-sphingomyelin (L/S) ratio and total phospholipid content. When the fat cell count was over 10%, the L/S ratio indicated maturity in all cases and the total phospholipids indicated maturity in all but 1 case. When the presence of blood or meconium in the amniotic fluid renders lung surfactant tests unreliable, an indication for the fat cell count exists.

Amniotic Fluid

Cell volume to aid analysis and technique of somatic cell counts in milk.

In conjunction with a Coulter Counter, somatic cells in milk were sized by electronic analysis. Quarter milk from cows with mastitis had a cell volume peak with a modal cell volume of 102 mu3 while milk from healthy quarters had no peak. Bulk milks with a peak had higher cell counts than milks where there was no peak. Dimensions of peak from bulk milks were the same as from quarter milks. Modal cell volume and cell count varied after milks were fixed with varying concentrations and types of fixative. The concentration of fixative recommended by the International Dairy Federation was sufficient, but only marginally so, and it should be increased.

Animals

The amniotic fluid foam test and fat cell count in malnourished and well-nourished fetuses.

A malnourished fetus has a higher concentration of pulmonary surfactant in the amniotic fluid than its well-nourished contemporary. The clinical value of this observation is discussed. Malnourished fetuses have higher "fat cell" counts in their amniotic fluid than well-nourished contemporaries. Well-nourished fetuses, at the same gestational age have high fat cell counts when the surfactant concentration is high, and low fat cell counts when the surfactant concentration is low. The relationship between surfactant concentration and the fat cell count is discussed.

Adipose Tissue

Enzymic methods for estimation of the somatic cell count in bovine milk. 1. Development of assay techniques and a study of their usefulness in evaluating the somatic cell content of milk.

Assay procedures were developed for a number of enzymes in milk which apparently originate from leucocytes. The enzymes studied were acid phosphatase, N-acetyl-beta-D-glucosaminidase, beta-glucuronidase, arylsulphatase, alpha-mannosidase, and catalase. Quarter-milk samples were analysed for enzyme activity and results compared with the electronic cell count and the Wisconsin Mastitis Test. All enzymes measured except acid phosphatase and alpha-mannosidase showed good correlation with the electronic cell count. Of the other 4 enzymes tested, beta-glucuronidase and arylsulphatase were unsuitable as diagnostic aids owing to the lengthy incubation periods required in their assay procedures. The assay of catalase, which involved the measurement of the initial rate of release of O2 using an O2 analyser apparatus, was rapid, sensitive and reasonably reliable, if fresh milk samples were used. The assay procedure for N-acetyl-beta-D-glucosaminidase was considered to be the most reliable, simple and rapid enzymic method for estimating the number of somatic cells in milk.

Acetylglucosaminidase

Improved automated optical somatic cell counting method for raw milk: collaborative study.

A collaborative study was conducted to compare the improved automated optical somatic cell counting procedure (OSCC II) with the direct microscopic somatic cell counting method (DMSCC) in raw milk. Samples were prefixed with formaldehyde and introduced into an Auto-Analyzer system. Dilution, clarification, and cell counting were performed automatically. Five collaborators participated in the study; they analyzed 48 samples in duplicate, using 2 different sampling rates. The results were compared with DMSCC counts reported by 3 different analysts. Statistical results show that the standard deviation for the DMSCC method is 0.0825 and for the OSCC II, at a sampling rate of 80/hr, 0.0434. When results are compared, the OSCC II procedure is as accurate as, and is significantly more precise than, the DMSCC method. The faster sampling rate of the OSCC II at 120/hr has some effect on precision but little effect on accuracy. The method has been adopted as official first action.

Animals

Mastitis history, California Mastitis Test, and somatic cell counts for identifying cows for treatment in a selective dry cow therapy program.

Mastitis history, California Mastitis Test scores, and filter-deoxyribonucleic acid cell counts from 232 cows were used to project the effectiveness of criteria which could be used to determine which cows to treat in a selective dry cow therapy program. Bacteriological analyses of quarter milk samples were used to identify infected cows. Effects of month, lactation parity, stage of lactation, and interaction of stage of lactation with lactation parity on somatic cell numbers in milk were determined. The various criteria singly or in combinations correctly identified from 50 to over 92% of the cows with one or more infected quarters at drying off. However, from 25 to 80% of the cows free of infection would have been treated as well. The most discriminating criteria were two monthly cell counts exceeding 500,000 cells per milliliter where 53% of the cows with infected quarters were identified correctly and only 25% of the uninfected cows would be treated, or the two monthly cell counts exceeding 500,000 cells per milliliter plus a California Mastitis Test score of +2 or +3 at drying off. Here 89% of the cows with infected quarters would be treated, and 56% of the uninfected cows would be treated.

Animals

Cell counts in the human cerebral cortex using a traditional and an automatic method.

This study compares the results of cell counting in the human cerebral cortex by a traditional and an automatic procedure. Four blocks were taken from standard areas of each of 10 brains from individuals aged 18-95 years and showing no neurological abnormality. Sections from each block were counted by traditional and automatic methods, both of which are described. The traditional method used photomicrographs and the automatic procedure used the quantimet 720 Image Analysing Computer. The degree of reproducibility of each method was measured and the results are discussed and tabulated. Comparison are made of the total cell counts, and the counts of cells measuring greater than 20 um across. The results show a high level of correlation between the two procedures but the manual count yields higher number for total cells and smaller numbers for large cells than the automatic method. The reasons for these discrepancies are discussed and examples of the arterfacts which produce them are giver. Using the traditional method described the amount of meterial examined took about 4 months to study. The sane tissue can be assessed in 1 day using the automatic apparatus. The high speed and close correlation achieved by the automatic procedure makes it a valuable aid in quantitative cell studies in the central nervous system.

Cell Count

A comparative study for the enumeration of peripheral blood white cell counts below 2.0 x 10(9)/1 using counting chambers and the Coulter Counter model "S".

Low white cell counts are performed in many laboratories by a visual method as the method of choice. In this study, the visual count has been compared to that obtained by the Coulter Model "S". The results obtained by the two methods show very close correlation, thus allowing the Coulter Model "S" to be used with confidence for all low white cell counts provided certain precautions are taken.

Humans