Effect of mitogenic and high, nonmitogenic concentrations of phytohemagglutinin and concanavalin A on the number of human lymphocytes in culture

A number of different investigators have observed a significant decrease in the number of lymphocytes in culture within 24 to 48 hr after the addition of mitogenic concentrations of phytohemagglutinin (PHA) and concanavalin A (Con A). In order to investigate the mechanism of this apparent decrease in cell number, we compared the number of cells in culture, determined by the pronase-cetrimide technique, to the amount of cell DNA and cell protein. Although cell number appeared to decrease following the addition of PHA or Con A in a dose-related fashion, cell DNA and cell protein did not change. When lymphocytes that were incubated with Con A were treated with alpha-methyl-D-mannoside prior to counting, a decrease in cell number or viability was not observed and there was close agreement between the number of cells and amount of cell DNA in culture. We conclude that the number of lymphocytes in culture does not decrease during the first 24 hr after the addition of mitogenic concentrations of PHA and Con A, and that previous reports to this effect may be accounted for by problems associated with counting lymphocytes that are agglutinated by mitogens.     

Effects of two types of inactivity on the number of white blood cells in rats

Prolonged inactivity is known to induce changes in responses of many physiological defense systems such as the hypothalamo-hypophyseal-adrenocortical axis, the sympathetic nervous system, and immuno-responsive systems. However, effects of various types of inactivity on immuno-responsive systems are still unknown. Therefore, the effects of two types of inactivity (immobilization: IMM and whole body suspension: WBS) on the number of white blood cells were studied in rats. Rats were divided into the control group and each inactivity group to compare the number of total white blood cells, lymphocytes, monocyte, neutrophil, eosinophil, and basophil during the experimental periods. Both IMM and WBS were maintained for 11 days. IMM markedly increased the number of total white blood cells, monocyte, neutrophil, and eosinophil in the 1st to 10th day. However, the number of total white blood cells, monocyte, neutrophil, and eosinophil during the experiment of WBS were characterized by the presence of a lag phase followed by the significant increased actions. IMM did not change the number of basophil during the experimental period. However, WBS increased the number of basophil in the 1st to 8th day to 2.8–4.8 times, compared with the values of the control. Both IMM and WBS did not change the number of lymphocytes. From these results, WBS increases the number of natural immunity cells without changing acquired immunity cells, and there are different responses in the number of total white blood cells, monocyte, neutrophil, eosinophil, and basophil between IMM and WBS.     

American College of Medical Genetics standards and guidelines for interpretation and reporting of postnatal constitutional copy number variants

Genomic microarrays used to assess DNA copy number are now recommended as first-tier tests for the postnatal evaluation of individuals with intellectual disability, autism spectrum disorders, and/or multiple congenital anomalies. Application of this technology has resulted in the discovery of widespread copy number variation in the human genome, both polymorphic variation in healthy individuals and novel pathogenic copy number imbalances. To assist clinical laboratories in the evaluation of copy number variants and to promote consistency in interpretation and reporting of genomic microarray results, the American College of Medical Genetics has developed the following professional guidelines for the interpretation and reporting of copy number variation. These guidelines apply primarily to evaluation of constitutional copy number variants detected in the postnatal setting.     

Principles of signal coding by the discharge pattern of a neuron population

The principles of information coding by rings of stochastic dependence, formed by neuron populations, are described. Such a population was shown to be capable of existing only in strictly definite stochastic states, determined by a certain number of rings of stochastic dependence, A system consisting of a fixed number of neurons was shown to be able to code and transmit a number of different messages equal to the square of the number of stochastic states permitted for that system. The number of messages differing from each other either in the number of letters or their order in the word was equal to the number of permitted stochastic states, whereas the number of messages containing the same number of letters in the word was equal to the difference between the number of permitted stochastic states and the number of neurons in the system. The alphabet consists of two letters: A for assembling of the ring of stochastic dependence, and B for breaking of this ring, together with punctuation signs indicating neither assembling nor breaking (compared with the initial state) of stochastic dependence rings.Translated from Fiziologicheskii Zhurnal SSSR imeni I. M. Sechenova, Vol. 70, No. 4, pp. 492–500, April, 1984.     

Quantitation of leukocytes in endomyocardial tissue from 100 normal human hearts at autopsy. Implications for diagnosis of myocarditis from biopsy specimens of living patients

From 100 normal human hearts, evenly distributed by age and sex, 5 endomyocardial samples were obtained from the septal surface of each ventricle with a cardiac bioptome. In each case, from both ventricles, the number of lymphocytes, eosinophils, plasma cells, and neutrophils was counted in 10 high-power (x400) microscopic fields, and the mean number of each type of leukocyte was calculated. In 95% of the 2,000 high-power fields, the number of interstitial lymphocytes was less than 5.0. Moreover, in all but one heart, the mean number of lymphocytes was less than 4.0. The mean number of eosinophils was 0.0-0.1 and of plasma cells was 0.0-0.3. The median number of neutrophils was 0.6. Recognition of the normal number of leukocytes in the heart may help to minimize false positive interpretations of myocarditis in biopsy specimens of endomyocardial tissue from living patients.     

Amplification of different ColE1 plasmids in an Escherichia coli relA strain

Amino acid starved cells of an E. coli relA strain accumulate a large amount of pBR322 plasmid DNA. In this study ColE1 related plasmids of different copy number and size including a high copy number plasmid mutant of pBR322 were amplified in a relA strain of E. coli K-12 under amino acid limitation in order to determine the upper plasmid level in amino acid starved cells. In all cases we measured a 4 to 6 fold increase of the plasmid copy number in comparison to log-phase cells independent of the size, the number of origins per plasmid molecule or the copy number in log-phase cells. The plasmid copy number in amino acid starved cells varies from about 200 (pBR322-dimer) to about 2000 (high copy number plasmid pERIII-BPL4, see Boros et al. 1986). Rop+ and rop- plasmids show the same amplification rate under the used conditions.     

Determination of nucleolar organizer activity in the differential diagnosis of severe dysplasia and cancer of the large intestine]

The method is based on the selective nucleolar staining with silver salts, the number of silver granules serves as an index of the nucleolar organizer activity which corresponds to the number of RNA-polymerases acting in the cell. The number of silver granules is 5.7, 6.3 and 6.8 per cell in mild, moderate and severe dysplasia, respectively. The histogram of the cell distribution, with regard to the silver grain number, in severe dysplasia is characterized by a shift to the left, and by one high peak. The average number of silver grains is significantly increased in non-invasive and invasive carcinoma and reaches the values of 18.8 and 20.4. A histogram in carcinoma is pulled to the right due to the cells with high content of silver grains and has many low peaks this reflecting variability of cells as to the silver grain content.     

A Dissociation of Number Meanings

We report a study of a patient with selectively preserved sequence meaning of numbers, but impaired access to cardinal number meaning. This limited understanding of number meaning was associated with a severe impairment in calculation tasks, with the inability to apply arithmetic principles and with an inverse distance effect in a number comparison task. In the remission process, better numerical understanding was reflected by better arithmetical competence and a normal distance effect in number comparison. The dissociation of different aspects of number meaning and the gradual development of arithmetical skills is well known in developmental psychology. The present case study suggests that a similar pattern can occur after cerebral lesions.     

Effects of extended cocaine conditioning in the reinstatement of place preference

Rats allowed extended access to cocaine self-administration develop a number of symptoms of addiction, such as greater susceptibility to drug-induced relapse. Using the conditioned place preference (CPP), the number of conditioning training sessions was increased in order to augment exposure to contextual cues associated with the effects of a drug. Mice were conditioned with a steady dose of 6 or 25 mg/kg of cocaine for 4, 8, 12, 16, 20 or 40 days. Weekly sessions of extinction followed the establishment of preference, after which a priming dose of cocaine was administered to reinstate the extinguished preference. The magnitude of the place preference effect was equal in all groups, independently of the number of conditioning sessions. The persistence of the place preference was not related with the number of sessions. Higher responsiveness to reinstatement of the extinguished preference occurred only with an intermediate number of conditioning sessions. In this way, the relation between the number of training sessions and vulnerability to relapse appeared to follow an inverted U-shaped function. Our results suggest that increasing the number of conditioning sessions from 12 to up to 16, without increasing the amount of drug administered, can be of great use in the study of vulnerability to relapse.     

Increase in the number of splenic plaque forming cells with length of gestation in untreated pregnant mice

The number of plaque forming cells (PFC) in spleens from untreated virgin and syngeneically pregnant CBA mice has been evaluated by use of two different hemolytic plaque assays. The total number of splenic IgM- and IgG-PFC in virgin and 16 day pregnant mice was determined by protein A-sheep red blood cell (SRBC) plaque assay. The number of both IgM- and IgG-PFC was significantly increased in spleens from pregnant mice, but the total number of IgM-PFC was several times higher than that of the IgG-PFC. In the same experiment the number of splenic anti-trinitrophenyl (TNP) PFC was determined simultaneously. The values obtained for anti-TNP PFC in the different animals were found to correlate with the values for IgM-PFC in the protein A-SRBC plaque assay. Finally, the number of splenic anti-TNP PFC was determined at different occasions during pregnancy (day 4, 8, 12, and 16). It was found that the number of anti-TNP PFC was elevated as early as the 4th day of pregnancy, and that the number of PFC per spleen increased with the length of gestation.     

Ploidy independence of Ag-NOR number in neuroblastoma.

BACKGROUND: Ag-NOR number and ploidy have potential use in predicting prognosis in malignancy. The issue of independence has not been conclusively studied. In contrast to other malignancies in neuroblastoma poor outcome is linked to diploidy, allowing independent analysis of the reaction of Ag-NOR status to outcome. EXPERIMENTAL DESIGN: A consecutive unselected series of 20 pediatric neuroblastoma patients was used to study the relation between ploidy and mean Ag-NOR number per nucleus. Ploidy was established by flow cytometric analysis of nuclear suspensions prepared from Formalin fixed and paraffin embedded tissue. The mean number of Ag-NORs per nucleus was determined using 3-microns paraffin sections, silver staining methods and quantification procedures detailed in the literature. RESULTS: The seven diploid lesions were found to have a mean Ag-NOR number per nucleus of 4.38 (range 0.86-7.5). The 13 aneuploid lesions were found to have a mean Ag-NOR number per nucleus of 2.31 (range 1.17-4.44). Probability of difference (Wilcoxon's two-tailed rank sum test) was greater than 99%. Nonsurvivors (6 of 10 diploid) had mean Ag-NOR number per nucleus of 4.34 (range 2.24-7.5). Survivors (9 of 10 aneuploid) had mean Ag-NOR number per nucleus of 1.73 (range 0.86-2.56). Probability of difference (Wilcoxon's two-tailed rank sum test) was greater than 99%. In aneuploid lesions (n = 13) a negative correlation exists between DNA index and mean number of Ag-NORs per nucleus (correlation coefficient 0.84, slope -0.74, probability of noncorrelation less than 0.010). CONCLUSIONS: The present observations offer the first independent support to the theory that Ag-NOR number is an independent variable to DNA ploidy based on the unique feature of neuroblastoma that diploid lesions have an established poorer prognosis than aneuploid lesion. The observations, although the number of cases is too limited for independent analysis, confirm previously published, strong negative relations between mean Ag-NOR number per cell and survival in childhood neuroblastoma. This relation is similar to findings in other solid tumor lesions.     

Mast cells and eosinophils are involved in activation of ulcerative colitis

PurposeThe intestinal mucosal immune cells such as the mast cells and eosinophils play an important role in the pathogenesis of ulcerative colitis (UC). The aim of present study was to compare the number of mast cells and eosinophils in patients with active and non-active ulcerative colitis. Another purpose was to found whether the number of eosinophils could correlate with number of mast cells in both tested groups.Material and MethodsThe twenty-five of formalin-fixed, paraffin-embedded tissue specimens of active ulcerative colitis, the twenty of non-active ulcerative colitis and the ten of controls were retrieved from archival material. Tryptase and chymase immunopositive cells were detected using immunohistochemical method. Additionally, the number of mast cells and eosinophils were detected using the most common histochemical methods.ResultsThe number of eosinophils and toluidine blue stained and tryptase immunopositive mast cells was significantly increased in active UC compared to non-active UC. In active stage of UC positive correlation between the number of mast cells stained with toluidine blue and the number of chymase and tryptase immunopositive mast cells were observed. Moreover, the number of eosinophils was significantly correlated with number of mast cells stained with toluidine blue and number of tryptase- and chymase immunopositive mast cells. In non-active stage of UC positive correlation was observed only between the number of mast cells stained with toluidine blue and chymase immunopositive cells and eosinophils.ConclusionsIn conclusion, our findings confirmed that mast cells and eosinophils are functionally involved in the course of UC.     

四君子汤复方总多糖对小鼠肠道粘膜相关淋巴组织的影响

目的：通过环磷酰胺所致小鼠肠道粘膜相关淋巴组织损伤模型研究四君子汤复方总多糖对肠道粘膜免疫的影响。方法：观察口服1g/kg四君子汤复方总多糖对100mg/kg的环磷酰胺处理24h后小鼠Peyer‘s结的数目、Peyer‘s结内细胞总数、CD3^ 、IgA^ 细胞数量和细胞凋亡的情况。结果：四君子汤复方总多糖可对抗环磷酰胺诱导的小鼠肠道粘膜相关淋巴组织中Peyer‘s结数目、Peyer‘s结细胞总数、肠粘膜内CD3^ 、IgA^ 细胞量的减少。细胞凋亡程度也明显减轻。结论：四君子汤复方总多糖可对抗环磷酰胺诱导的小鼠肠粘膜相关淋巴组织损伤,提示多糖可能对肠道粘膜免疫具有改善作用。     

Suppression of apoptosis in hepatocytes by fructose-modified dendrimers.

By immobilizing fructose-modified dendrimers on a polystyrene culture plate, the number of initially adhered hepatocytes on it was increased. Moreover, increasing the number of generations of fructose-modified dendrimer (fructose-dendrimer) increased the number. Urea synthesis per unit area also was increased, corresponding to the increase in the number of initially adhered hepatocytes. This result suggests that the fructose-dendrimers do not cause a decline in cell function. On the other hand, apoptosis of hepatocytes occurs during cultivation, and results in a decrease in the number of adhered cells and a decline in cell function. Fructose-dendrimers were found to suppress apoptosis of hepatocytes. This characteristic is considered to be responsible for the increase in the number of initially adhered hepatocytes without a decline in cell function. Fructose-dendrimers are shown to be very suitable scaffolds for use in a high-performance bioartificial liver support system.     

Correlation of organism burden and alveolar macrophage counts during infection with Pneumocystis carinii and recovery

Changes in the number of alveolar macrophages were correlated with organism burden during Pneumocystis carinii infection. The lungs of healthy, dexamethasone-treated, and dexamethasone-treated and P. carinii-infected rats were lavaged with phosphate-buffered saline. Counting of alveolar macrophages in the lavage fluids revealed that P. carinii infection caused a 58% decrease in the number of alveolar macrophages and that higher P. carinii organism burdens caused a more rapid decrease in alveolar macrophage number. As a control, healthy rats were challenged with the same number of organisms as that normally used to generate P. carinii infections in dexamethasone-treated rats. Thirteen days after challenge, these rats had a profound (54%) increase in alveolar macrophage number in response to the challenge, while the number of alveolar macrophages in immunosuppressed and P. carinii-infected rats had decreased significantly by this time point. These experiments created the first animal model to mimic human pneumocystis pneumonia in alveolar macrophage number alterations. Reduction of P. carinii organism numbers by treatment of rats with trimethoprim and sulfamethoxazole brought a slow rebound in alveolar macrophage number, while recovery from P. carinii infection by cessation of immunosuppression brought a rapid rebound in alveolar macrophage number. These results suggest that both the immune state of the host and P. carinii burden affect alveolar macrophage number.     

Estimated number of off‐target candidate sites for antisense oligonucleotides in human mRNA sequences

Antisense oligonucleotide (ASO) therapeutics are single‐stranded oligonucleotides which bind to RNA through sequence‐specific Watson–Crick base pairings. A unique mechanism of toxicity for ASOs is hybridization‐dependent off‐target effects that can potentially occur due to the binding of ASOs to complementary regions of unintended RNAs. To reduce the off‐target effects of ASOs, it would be useful to know the approximate number of complementary regions of ASOs, or off‐target candidate sites of ASOs, of a given oligonucleotide length and complementarity with their target RNAs. However, the theoretical number of complementary regions with mismatches has not been reported to date. In this study, we estimated the general number of complementary regions of ASOs with mismatches in human mRNA sequences by mathematical calculation and in silico analysis using several thousand hypothetical ASOs. By comparing the theoretical number of complementary regions estimated by mathematical calculation to the actual number obtained by in silico analysis, we found that the number of complementary regions of ASOs could be broadly estimated by the theoretical number calculated mathematically. Our analysis showed that the number of complementary regions increases dramatically as the number of tolerated mismatches increases, highlighting the need for expression analysis of such genes to assess the safety of ASOs.     

Characterization of copy number-stable regions in the human genome

In the past few years the number of copy number variants (CNVs) identified in the human genome has increased significantly, but our understanding of the functional impact of CNVs is still limited. Clinically significant variations cannot easily be distinguished from benign, complicating interpretation of patient data. Multiple studies have focused on analysis of regions that vary in copy number in specific disorders. Here we use the opposite strategy and focus our analysis on regions that never seem to vary in the general population, hypothesizing that these are copy number stable because variations within them are deleterious. Our results show that copy number stable regions are characterized by correlation with a number of genomic features, allowing us to define a list of genomic regions that are dosage sensitive in humans. We find that these dosage-sensitive regions show significant overlap with de novo CNVs identified in patients with intellectual disability or autism. There is also a significant association between copy number stable regions and rare inherited variants in autism patients, but not in controls. Based on this predictive power, we propose that copy number stable regions can be used to complement maps of known CNVs to facilitate interpretation of patient data.     

Effect of irradiation,cyclophosphamide, and etoposide (VP-16) on number of peripheral blood and peritoneal leukocytes in mice under normal conditions and during acute inflammatory reaction

In order to develop a suitable model for studying the role of granulocytes and monocytes in resistance against pathogenic microorganisms, we investigated the effect of irradiation and cytostatic treatment (cyclophosphamide and VP-16) on the number of both peripheral blood and peritoneal leukocytes in male Swiss mice. Irradiation and cyclophosphamide treatment severely decreased the number of both granulocytes and monocytes in peripheral blood, whereas VP-16 only lowered the number of blood monocytes to a significant degree and had little effect on the number of blood granulocytes or lymphocytes. When normal mice were injected intraperitoneally with newborn calf serum (NBCS) the number of peritoneal granulocytes rose about 100-fold within 6 h. In irradiated and cyclophosphamide-treated mice, this influx of granulocytes into the peritoneal cavity was virtually eliminated, as was the concomitant increase in the number of blood granulocytes; in VP-16-treated mice, on the other hand, the number of peripheral blood and peritoneal granulocytes increased to the same degree as in normal mice. An increase in the number of peripheral blood monocytes and peritoneal macrophages occurred 24–48 h after injection of NBCS in normal mice. This increase was significantly impaired by irradiation as well as by treatment with cyclophosphamide or VP-16.