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PubMedDate not supplied

Prestimulation of head and neck cancer cells with growth factors enhances treatment efficacy.

OBJECTIVES: In recent years, new chemotherapy regimens with promising activity, especially in first-line therapy (induction chemotherapy) of head and neck cancer (SCCHN), have been developed. Nevertheless, a major problem concerning the response of SCCHN to chemotherapy is the high percentage of resting cells (G0-phase cells) being resistant to chemotherapy. To overcome this phenomenon, the capacity of several cytokines to switch on cells into the division cycle and progress to the chemosensitive phases (S-, M-phases) was investigated. MATERIALS AND METHODS: Interleukin-6, serotonin, granulocyte colony stimulating factor (G-CSF) and epidermal growth factor (EGF) were used to stimulate G0-phase squamous cell cancer cells (Detroit 562, A431, UM-SCC 10B) for re-entry into the cell cycle to enhance the response to cisplatin. The proportion of G0-phase cells was detected through multicolor FACS analysis and Ki-67 staining. RESULTS: Cell cycle re-entry was most effective after combination treatment with serotonin + EGF. The proportion of G0-phase cells was significantly reduced after stimulation with serotonin + EGF (p < 0.05). Corresponding to cell cycle re-entry, the cytotoxic effect of cisplatin was significantly (p < 0.04) enhanced in the prestimulated compared to the control cells (cisplatin mono-treatment). CONCLUSION: Our investigations demonstrated for the first time that sensitizing G0-phase squamous cell carcinoma cells for chemotherapy is possible by prestimulation with target cytokines. Considering that up to 95% of tumor cells are in the resting (G0) phase of the cell cycle at the initiation of chemotherapy, prestimulation with EGF and serotonin could contribute to a synchronization of cancer cells. This would clearly enhance the cytotoxic effect.

Antineoplastic Agents↗
PubMed2006

A cytogenetic analysis in two cases of malignant peripheral nerve sheath tumor showing hypodiploid karyotype.

In this study, we report cytogenetic findings in two cases of malignant peripheral nerve sheath tumor (MPNST) with hypodiploid karyotypes. A G-band technique, multicolor fluorescence in situ hybridization (m-FISH) and comparative genomic hybridization (CGH) were used and compared in this investigation. In both tumors, the G-band and m-FISH analysis demonstrated multiple rearrangements on chromosomes 1-5, 8-12, 15-17, 20 and 21, whereas CGH exhibited gains at 8q and 4q. Both of the structural aberrations and the genomic imbalances of the chromosomes may play an important role in the pathogenesis and development of MPNST. No cytogenetic abnormalities specific for MPNST were found in the present cases or in other previously reported cases. This may reflect the diversity or heterogeneity of MPNST that exhibit various clinical and histological features. However, there are few cases described in detail on a morphologic pattern of MPNST, a correlation between the cytogenetic aberrations and the histologic patterns are still uncertain.

Aged↗
PubMed2003

[Molecular cytopathology of lung cancer--prevalence of genetic alterations and their role in the development of molecular biomarkers].

Carcinogenesis of lung cancer proceeds via a complex process that involves multiple genetic abnormalities, which do not necessarily have a linear progression. Genetic alterations include aneuploidy, deletions and amplifications of chromosomal regions, loss of heterozygosity (LOH), microsatellite alterations, point mutations and aberrant promoter methylation. There is considerable effort to use these genetic alterations as molecular biomarkers for early cancer diagnosis applying different approaches. An ideal tumor marker should be highly sensitive, tumor specific, easily to handle and non-cost intensive. While previous studies used screening for mutations, LOH and microsatellite alterations, more recent strategies concentrate on multicolor fluorescence in situ hybridization (FISH) and aberrant promoter methylation. Since in general the genetic alterations are prone to be more extensive in tumor cells as compared to non-tumor cells, methods that provide quantitative data (e.g., methylation specific real-time PCR) are likely to improve specificity. Consequently, molecular biomarkers could constribute to a more accurate risk assessment in carcinogen exposed individuals and early molecular cytologic diagnosis of precancerous lesions and cancers of the lung.

Allelic Imbalance↗
PubMed1990

Colony formation of clone-sorted human hematopoietic progenitors.

To characterize human hematopoietic progenitors, we performed methylcellulose cultures of single cells isolated from a population of CD34+ cells by fluorescence-activated cell-sorting (FACS) clone-sorting system. CD34+ cells were detected in bone marrow (BM) and peripheral blood (PB) cells at incidences of 1.0% and 0.01% of total mononuclear cells, respectively. Single cell cultures revealed that approximately 37% of BM CD34+ cells formed colonies in the presence of phytohemagglutinin-leukocyte conditioned medium and erythropoietin. Erythroid bursts-, granulocyte-macrophage (GM) colony-, and pure macrophage (Mac) colony-forming cells were 10% each in CD34+ cells. Approximately 15% of PB CD34+ cells formed colonies in which erythroid bursts were predominant. CD34+ cells were heterogeneous and fractionated by several antibodies in FACS multicolor analysis. In these fractionated cells, CD34+, CD33+ cells formed GM and Mac colonies 7 to 10 times as often as CD34+, CD33- cells. Most of the erythroid bursts and colonies were observed in the fraction of CD34+, CD13- cells or CD34+, CD33- cells. The expression of HLA-DR on CD34+ cells was not related to the incidence, size, or type of colonies. There was no difference in the phenotypical heterogeneity of CD34+ cells between BM and PB. About 10% of CD34+ cells were able to form G colonies in response to granulocyte colony-stimulating factor (G-CSF) and to form Mac colonies in GM-CSF or interleukin-3 (IL-3). Progenitors capable of generating colonies by stimulation of G-CSF were more enriched in CD34+, CD33+ fraction than in CD34+, CD33- fraction. Thus, single cell cultures using the FACS clone-sorting system provide an accurate estimation of hematopoietic progenitors and an assay system for direct action of colony-stimulating factors.

Antigens, CD34↗
PubMed2006

[Effects of rhG-CSF mobilization on immunological properties of grafts from peripheral blood and bone marrow].

This study was aimed to investigate the difference of immunological properties between recombination human granulocyte colony-stimulating factor (rhG-CSF) mobilized peripheral blood grafts (G-PB) and rhG-CSF primed bone marrow grafts (G-BM). The lymphocyte proliferation ability and the quantities of interleukin-4 (IL-4) and interferon-gamma (IFN-gamma) secreted by T cells were determined by using MTT assays and sandwich ELISA; T cell subgroups, dendritic cells (DC), monocytes and the expression of CD28 costimulatory molecules on T cells were determined by multicolor flow cytometry. The results showed that the absolute numbers of lymphocytes, monocytes, CD3+, CD4+ and CD8+ T cells as well as DC1 and DC2, the ratios of CD4/CD8 in G-PB were significantly higher than those in G-BM, respectively (P < 0.001). T cell proliferation ability was significantly higher in G-PB than that in G-BM (P < 0.05). The quantities of IFN-gamma and IL-4 secreted by T cells per microliter of G-PB was significantly higher than those of G-BM, the ratios of IL-4/IFN-gamma were significantly lower in G-PB than that in G-BM (P < 0.001). As compared with G-BM, the ratio between DC2 and T-lymphocyte was significantly low in G-PB (P < 0.01), whereas the percentage and overall expression of CD28 on CD4+ and CD8+ cells were significantly high in G-PB (P < 0.05). It is concluded that T cell hyporesponsiveness of G-PB and G-BM induced by rhG-CSF in vivo were confirmed to be different, and the difference of immunological properties between G-PB and G-BM may explain the lower incidence of GVHD and lower relapse after G-BM and G-PB transplantation respectively.

Adolescent↗
PubMed2006

[Cytogenetic disorders in chronic B-cell lymphoid leukemia and their relations with clinicobiological features and prognosis of the disease].

AIM: To study a relationship between cytogenetic disorders, clinicobiological characteristics and prognosis in chronic B-cell lymphoid leukemia (B-CLL). MATERIAL AND METHODS: Cytogenetic examination of blood, bone marrow and lymph node cells from 135 patients (90 males and 45 females aged 23-84 years) with chronic B-CLL was made. The patients were followed up from 1 month to 25 years. Before the cytogenetic examination specific therapy was not given. B-CLL was staged by K. Rai, forms--by A.L. Vorobyev and M.D. Brilliant. All the patients have undergone standard cytogenetic examination, FISH with multicolor probe to loci with possible frequent aberrations (del3q14, del11q23, del17p13, trisomia 12), determination of CD38 antigen expression on circulating tumor cells. Mutation status of the genes of immunoglobulins variable region (IgVH) was defined in 61 patients. RESULTS: Del13q14 was detected in 34 cases, del11q23--in 26, trisomia of chromosome 12--in 17 cases, del 17p13--in 8, absence of q-arm of chromosome 13--in 3 cases. 61 patients had no karyotype defects. Three prognostic groups of the patients were identified: favourable prognosis--patients without disorders of karyotype and one chromosomal aberration--del13q14; intermediate prognosis patients with dell1q23 and trisomia of chromosome 12; poor prognosis--patients with del17p13 and complex disorders of karyotype. CONCLUSION. Cytogenetic study help determine prognosis of B-CLL and detect patients in need of early therapy.

ADP-ribosyl Cyclase 1↗
PubMedDate not supplied

[Characterization of marker chromosomes using molecular cytogenetic methods in patients with mental retardation and congenital malformations].

INTRODUCTION: Until recently, great variety of marker chromosomes and difficulties with their identification have presented a problem for cytogenetic and clinical interpretation of the karyotype. At present, molecular cytogenetic methods of chromosome analysis enable precise characterization of such abnormalities providing knowledge necessary for estimation of their genetic risk. AIM: The aim of the study was molecular cytogenetic characterization of marker chromosomes recognized in three patients, an analysis of clinical features in relation to the abnormality and estimation of genetic risk of identified markers. PATIENTS AND METHODS: Karyotypes of three phenotypically abnormal patients were estimated in lymphocytes from peripheral blood by G banding analysis. Marker chromosomes were identified by fluorescence in situ hybridization (FISH), multiplex FISH, multicolor band and high resolution comparative genomic hybridization methods. RESULTS: Marker chromosomes were identified as inv dup(22)(pter->q11.2::q11.2->pter), der(8)(:p22->q11.2:), der(2l)(:pter->q21.3:) and der(19)(:p11->q13.1). All of them contained euchromatic sequences. First marker, an inverted duplication of chromosome 22q11.2 corresponding to tetrasomy of this chromosome region was recognized in a child with partial cat eye syndrome. Two further markers derived from chromosomes 8 and 21 were found in a child with mosaic karyotype and clinical features of trisomy 8p. In the third case additional chromosome material was derived from chromosome 19 and it was found in a patient with mild mental retardation and clinical features of ovary dysgenesis. Genetic risk of identified marker chromosomes except for mar(19) was estimated as high. CONCLUSIONS: Our results provide further evidence for diagnostic value of molecular cytogenetic methods. They also confirmed the general opinion of the high risk of phenotypic abnormalities in the carriers of marker chromosomes containing euchromatic sequences.

Abnormalities, Multiple↗
PubMed1991

Phenotypic complexity of intraepithelial lymphocytes of the small intestine.

A detailed phenotypic analysis of intraepithelial lymphocytes (IEL) of the small intestine was performed using multicolor fluorescence flow cytometry. CD4+8+ IEL (double positives; DP) could be detected in significant numbers in preparations from several mouse strains. DP IEL expressed Tcr alpha, beta and Thy1. Comparison of Tcr alpha, beta levels of thymocytes and IEL revealed that whereas the majority of DP thymocytes expressed low Tcr levels, DP IEL expressed high, mature T cell levels of Tcr. In addition, DP IEL were generally Ly3- (CD8 beta), unlike their thymic counterparts, which are Ly3+. Ly3 was not present on Tcr gamma, delta IEL, whereas CD4-8+ Tcr alpha, beta IEL contained Ly3- and Ly3+ subsets. The Ly3- population in either Tcr-bearing subset could be further subdivided by Thy1 expression. Ly1 (CD5) expression was also examined, and none of the Tcr gamma, delta IEL were Ly1+. Based on Thy1, Ly3, and Ly1 expression, four CD4-8+ Tcr alpha, beta IEL subsets were detected. The results indicate the cellular complexity of the IEL compartment rivals that found in the thymus. These findings are discussed in light of recent data suggesting an extrathymic origin of some IEL.

Animals↗
PubMed1996

The ecological characteristics of Anopheles mosquitoes associated with low malaria transmission in Siwa oasis, Egypt.

The ecology of Anopheles sergentii and An. multicolor was investigated over eight months to examine the entomological factors maintaining the low Plasmodium vivax transmission in Siwa oasis. The two species were encountered as larvae while the dominant species, Anopheles sergentii was also collected as adults. Breeding sites were characterized for the two species in six localities. Larvae of both species were common during June and November. In Bahi El Din as an indicator village with a higher endemicity, An. sergentii females were endophilic but more common inside animal sheds than in houses, attracted more to donkey-baited traps than to human hosts, more endophagic as human biters and were more common in May/June and November. Female survivorship was higher in November than in the other months. Vector potential of An. sergentii was predicted and revealed that the species characteristics (low human contact, short survival and zoophilic feeding behavior) are responsible for maintaining such low and unstable malaria in Siwa oasis.

Animals↗
PubMed1999

Bionomics of the anopheline vectors in an endemic area in Fayoum Governorate, Egypt.

A one-year longitudinal entomological study was carried out at Kafr Fazara village, Sinnuris District, Faiyoum Governorate from January to December 1996. The results revealed that three species of anophelines were found. A. sergenti was the most prevalent species followed by A. multicolor and the least prevalent one was A. pharoensis. The influence of climatic conditions on anopheline abundance was also studied. The mean monthly temperature has a significant effect on larvae abundance, while no significant correlation was found concerning relative humidity or wind speed for both larvae and adult anophelines. The transmission season of P. falciparum in Faiyoum Governorate extended to more than eight months a year which could explain the persistence of malaria up there. The bionomics of the recorded vectors were discussed.

Analysis of Variance↗
PubMed2006

Genetic polymorphisms of human ABC transporter ABCG2: development of the standard method for functional validation of SNPs by using the Flp recombinase system.

The vector-mediated introduction of cDNA into mammalian cells by calcium phosphate co-precipitation or permeation with lipofectamine is widely used for the integration of cDNA into genomic DNA. However, integration of cDNA into the host's chromosomal DNA occurs randomly at unpredictable sites, and the number of integrated recombinant DNAs is not controllable. To investigate the effect of genetic polymorphisms of ABCG2 on the protein expression and the drug resistance profile, we developed the Flp-In method to integrate one single copy of ABCG2 variant-cDNA into FRT-tagged genomic DNA. More than 20 metaphase spreads were examined for both fluorescence in situ hybridization (FISH) mapping and multicolor-FISH analysis, and it has been revealed that ABCG2 cDNA was incorporated into the telomeric region of the short arm on one of chromosomes 12 in Flp-In-293 cells. Based on the currently available SNP data for human ABCG2, we have created a total of seven variants by site-directed mutagenesis and stably expressed them in Flp-In-293 cells. While mRNAs of those integrated ABCG2 variants and wild type were evenly expressed in Flp-In-293 cells, the protein expression levels of F208S and S441N variants were found to be markedly low. It is suggested that the protein instability due to enhanced degradation resulted in the low levels of their protein expression. Thus, the Flp recombinase system would provide a useful tool to validate the effect of nonsynonymous SNPs on the protein stability and post-translational modification of ABCG2.

ATP Binding Cassette Transporter, Subfamily G, Mem↗
PubMed1991

Clinical features of dysplastic nevi.

The clinical features of 100 dysplastic nevi were tabulated. Although certain characteristics were present in most or all of these melanocytic nevi, there was a marked heterogeneity of other clinical features. The preponderant type of large (greater than or equal to 8 mm) melanocytic nevus in patients with classic dysplastic nevi is a papule or plaque with the following characteristics: multicoloration (various shades of tans, browns, reds, or black); slightly raised height for its broad diameter; mamillated surface; and lack of hypertrichosis. An atlas illustrates some of the clinical varieties of melanocytic nevi in this syndrome.

Dysplastic Nevus Syndrome↗
PubMed1990

Differences in the expression profiles of CD45RB, Pgp-1, and 3G11 membrane antigens and in the patterns of lymphokine secretion by splenic CD4+ T cells from young and aged mice.

Previous studies indicate that the 3G11, CD45RB, and Pgp-1 determinants are differentially expressed on CD4+ T cell subsets in the mouse. We used multicolor immunofluorescence staining and flow cytofluorometric analysis to examine the expression of each of these determinants on splenic CD4+ cells from young (age 3 to 6 mo) and aged (age 24 to 26 mo) C57BL/6 mice. The CD4+ pool from aged mice contained significantly reduced numbers of 3G11+ and CD45RBhi cells, but increased numbers of Pgp-1hi cells, in comparison with the young group. Analysis of the simultaneous expression of all three subset determinants on CD4+ cells revealed that, in young mice, the major fraction (greater than 50%) was 3G11+CD45RBhiPgp-1lo. Among the less prevalent cell phenotypes, reductions in 3G11 expression correlated with decreases in CD45RB levels and increases in Pgp-1 levels. The phenotype that dominated the young group (3G11+CD45RBhiPgp-1lo) was approximately fivefold less represented in the aged group. The CD4+ pool from aged mice was characterized by increases in the 3G11-CD45RBvariablePgp-1hi and the 3G11+CD45RBloPgp-1hi phenotypes. To evaluate possible age-associated differences in cytokine secretion patterns by splenic CD4+ cells, purified CD4+ cells from each age group were stimulated in vitro with immobilized anti-CD3 epsilon mAb and accessory cells. At various times thereafter, supernatants from cultures were tested for IL-2 and IL-4 content by using the CTLL.6 and 11.6 bioassays, respectively, and the CD4+ cells were assayed for [3H]TdR uptake. Cell cultures from the aged group exhibited similar peak IL-2 accumulation and lower peak [3H]TdR uptake, but greatly increased peak IL-4 accumulation, as compared with cell cultures from the young group. The expression patterns of subset determinants, in conjunction with cytokine secretion profiles, indicate that, in aged mice, marked alterations occur in the subset composition of the splenic CD4+ cell pool. These findings are discussed in the context of previous findings on changes in T cell reactivity with advancing donor age.

Aging↗
PubMed1991

A direct demonstration of functional specialization in human visual cortex.

We have used positron emission tomography (PET), which measures regional cerebral blood flow (rCBF), to demonstrate directly the specialization of function in the normal human visual cortex. A novel technique, statistical parametric mapping, was used to detect foci of significant change in cerebral blood flow within the prestriate cortex, in order to localize those parts involved in the perception of color and visual motion. For color, we stimulated the subjects with a multicolored abstract display containing no recognizable objects (Land color Mondrian) and contrasted the resulting blood flow maps with those obtained when subjects viewed an identical display consisting of equiluminous shades of gray. The comparison identified a unique area (area V4) located in the lingual and fusiform gyri of the prestriate cortex. For motion, blood flow maps when subjects viewed moving or stationary black and white random-square patterns were contrasted. The comparison identified a unique area located in the region of the temporo-parieto-occipital junction (area V5). We thus provide direct evidence to show that, just as in the macaque monkey, different areas of the human prestriate visual cortex are specialized for different attributes of vision. The striate cortex (V1) and the contiguous visual area (V2), which in the monkey brain feed both the homologous areas, were active in all 4 conditions. This pattern of activity allowed us to use an extension of the approach to assess the functional relationship between the 3 areas during color and motion stimulation. This is based on an hypothesis-led analysis of the covariance structure of the blood flow maps and promises to be a powerful tool for inferring anatomical pathways in the normal human brain.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1990

Factors affecting the human-feeding behavior of anopheline mosquitoes in Egyptian oases.

Blood meals were tested by a direct enzyme-linked immunosorbent assay (ELISA) for 424 Anopheles sergentii and for 63 An. multicolor collected in Siwa, Farafra and Bahariya oases in the Western Desert of Egypt. Both species were highly zoophilic. Human blood-feeding by An. sergentii was less common in Bahariya (2.3%) and Farafra (1.3%) than in Siwa (15.3%). A likely explanation is that large domestic animals are held at night inside houses in Bahariya and in Farafra whereas in Siwa, animals are usually housed outdoors in sheds. These patterns of An. sergentii human-feeding behavior may contribute to the persistence of low-level Plasmodium vivax transmission in Siwa in contrast to negligible or no transmission in Bahariya and Farafra.

Animal Husbandry↗
PubMed1989

Expression of B cell surface receptors. II. IL-4 can accelerate the developmental expression of the murine B cell IgE Fc receptor.

The ability of IL-4 to influence the developmental expression of the murine B cell IgE Fc receptor (Fc epsilon R) was examined. Spleen cells from neonatal mice of increasing age were incubated overnight with IL-4 and subsequently examined with multicolor flow cytometry. The results demonstrate that IL-4 can significantly increase the number of maturing B cells which express the Fc epsilon R. This effect was only seen however, on those neonatal B cells which already displayed surface IgD. Splenic B cells which were IgM+, IgD- failed to express the Fc epsilon R when treated with IL-4, even though they responded by increasing their level of class II Ag expression. Further experiments showed that the inability of IgD- immature B cells to express the Fc epsilon R could not be entirely explained by their assignment to the Ly-1 lineage. Taken together, these results indicate that IL-4 can accelerate the developmental expression of the B cell Fc epsilon R, but only on those B cells that are mature enough to express IgD.

Animals↗
PubMed1989

Bagolini lenses vs. PFL filters in the diagnosis of correspondence.

This study introduces a new product, the PFL-7501 filter (prism linear filter), as a test for correspondence. Thirty consecutive strabismic subjects were asked to verbally describe, then draw, the images seen from a small light source through the Bagolini #4 striated lenses and the PFL-7501 filters. A 97% concordance was found between the two materials. The bright, multicolored streaks seen through the PFL filters were easier for all subjects to see and describe than the images seen through the Bagolini striated lenses.

Adolescent↗
PubMed1986

Anopheles population dynamics in two malaria endemic villages in Faiyum Governorate, Egypt.

Anopheles populations were monitored for one year in 2 neighboring villages in Faiyum Governorate, Egypt, to study factors causing differences in malaria prevalence. Both villages contained the following species: Anopheles pharoensis, An. sergentii, An. multicolor and An. tenebrosus. Abundant larval breeding sites in Abheet, the village with the higher malaria rate, accounted for the higher adult densities observed from human and animal biting collections and from indoor resting collections. Anopheles pharoensis and An. sergentii were the dominant species in Abheet with seasonal biting activity extending from May to December, reaching a peak in November. In El Zawya, the village with the lower malaria rate, An. pharoensis dominated, reaching seasonal peaks in June and August. Inside houses, An. sergentii was common from May to January in Abheet, but rare in El Zawya. Anopheles pharoensis and An. sergentii were both incriminated as malaria vectors based upon their seasonal abundance and the finding of sporozoite positive specimens during the peak malaria season.

Animals↗