Effects of age, HIV, and HIV-associated clinical factors on neuropsychological functioning and brain regional volume in HIV+ patients on effective treatment.

It is yet unclear if people infected with human immunodeficiency virus (HIV+) on stable, combined antiretroviral therapies (cARTs) decline with age at the same or greater rate than healthy people. In this study, we examined independent and interactive effects of HIV, age, and HIV-related clinical parameters on neuropsychological functioning and brain regional volume in a sizable group of Polish HIV+ men receiving cART. We also estimated the impact of nadir CD4 cell count, CD4 cell count during participation in the study, duration of HIV infection, or duration of cART along with age. Ninety-one HIV+ and 95 control (HIV-) volunteers ages 23-75 completed a battery of neuropsychological tests, and 54 HIV+ and 62 HIV- of these volunteers participated in a brain imaging assessment. Regional brain volume in the cortical and subcortical regions was measured using voxel-based morphometry. We have found that HIV and older age were independently related to lower attention, working memory, nonverbal fluency, and visuomotor dexterity. Older age but not HIV was associated with less volume in several cortical and subcortical brain regions. In the oldest HIV+ participants, age had a moderating effect on the relationship between the duration of cART and visuomotor performance, such as that older age decreased speed of visuomotor performance along with every year on cART. Such results may reflect the efficacy of cART in preventing HIV-associated brain damage. They also highlight the importance of monitoring neuropsychological functioning and brain structure in HIV+ patients. This is particularly important in older patients with long adherence to cART.

Interlaboratory Comparison of Six Real-Time PCR Assays for Detection of Bovine Leukemia Virus Proviral DNA.

Quantitative real-time PCR (qPCR) is increasingly being used for the detection of bovine leukemia virus (BLV) proviral DNA. Nevertheless, quality control for the validation and standardization of such tests is currently lacking. Therefore, the present study was initiated by three Office International des Epizooties (OIE) reference laboratories and three collaborating laboratories to measure the interlaboratory variability of six already developed and available BLV qPCR assays. For that purpose, an international panel of 58 DNA samples reflecting the dynamic range of the majority of the assays was distributed to six testing centers. Based on qualitative results, the overall agreement among all six laboratories was moderate. However, significant variability in the measurement of the BLV proviral DNA copy number was observed among different laboratories. Quantitative PCR assays, even when performed by experienced staff, can yield large variability in BLV proviral DNA copy numbers without harmonization. Further standardization of different factors (i.e., utilization of unified protocols and unique calibrators) should increase interlaboratory agreement.

Determination of cytokine profiles in populations of dendritic cells from cattle infected with bovine leukaemia virus.

Dendritic cells (DCs) due to their ability to present antigens are essential during the immune response to infections. The aim of the study was to evaluate the influence of bovine leukaemia virus (BLV) infection on DC properties. Cytokine profiles of myeloid, plasmacytoid and mono- cyte derived DCs from BLV infected cattle were analysed. Concentrations of IL-6, IL-10, IL-12, IFN-γ, and TNF-α in DC cultures were measured by flow cytometry. Obtained results indicated activation of pDCs population, where a significant increase in production of the IFN-γ was shown. Meanwhile, a decrease in production of IFN-γ and increase in production of IL-10 were shown in mDCs; the main population responsible for antigens presentation. This may indicate a contribu- tory role of the population during the process of persistent infection. In MoDCs population a significant elevation in secretion of proinflammatory cytokines - IL-6 and TNF-α was noted.

Microbiological quality of selected spices and herbs including the presence of Cronobacter spp.

The cultivation of spices and herbs in parts of the world characterized by warm climate and high humidity provides excellent conditions for the development of microorganisms, including the undesirable ones. The aim of this study was to determine the microbiological quality of spices and herbs available on the Polish market, considering the occurrence of Cronobacter species bacteria. Analyses covered 60 samples of commercial spices and herbs, including 38 samples of dried herbs (basil, bay leaves, thyme, oregano, tarragon, marjoram, dill, parsley, rosemary, lovage) and 16 samples of seasoning blends as well as 6 samples of spices seeds and fruits (pimento, black pepper, coriander). All samples were tested for the total count of aerobic mesophilic bacteria (TAMB) and for the presence of Cronobacter spp. In most of the samples of spices and herbs (60.0%), the TAMB did not exceed 10(4) CFU/g, and the level regarded as unacceptable (>10(6) CFU/g) was not identified in any of the samples. The presence of Cronobacter spp. was demonstrated in 10 (16.7%) samples of the analyzed products, however these were mainly samples of herbs (basil, tarragon, parsley) and one sample of a seasoning blend (Provence herbs). The highest microbiological contamination (TAMB) was found in samples of herbs (oregano, tarragon, basil) and in ready seasoning blends, in 21.1% and 25.0% of which the total count of aerobic mesophiles was in the range of 10(5)-10(6) CFU/g. In all samples of spices seeds and fruits (coriander, black pepper and pimento), the total count of aerobic bacteria reached <10(4) CFU/g. Results achieved in the study indicate good hygienic conditions in the production process of spices and herbs available on the Polish market. The study demonstrated also that dried spices and herbs may be carriers of Cronobacter species bacteria, though their presence in not often detected in products of this type.

Study on application of biocellulose-based material for cheese packaging.

Bacterial cellulose (BC, biocellulose) is a natural polymer of microbiological origin that meets the criteria of a biomaterial for food packaging. The aim of the research was to obtain biocellulose and test its chemical as well as physical characterization as a potential packaging for Dutch-type cheeses. Four variants of biocellulose-based material were obtained: not grinded and grinded variants obtained from YPM medium (YPM-BCNG and YPM-BCG, respectively) and not grinded and grinded variants from acid whey (AW) (AW-BCNG and AW-BCG, respectively). It was demonstrated that AW-BCNG exhibited the highest thermostability and the highest degradation temperature (348 °C). YPM-BCG and YPM-BCNG demonstrated higher sorption properties (approx. 40 %) compared to AW-BCG and AW-BCNG (approx. 15 %). Cheese packaged in biocellulose (except for YPM-BCNG) did not differ in water, fat, or protein content compared to the control cheese. All of the biocellulose packaging variants provided the cheeses with protection against unfavourable microflora. It was demonstrated that cheeses packaged in biocellulose were characterized by lower hardness, fracturability, gumminess, and chewiness than the control cheese sample. The results obtained indicate that BC may be a suitable packaging material for ripening cheeses, which shows a positive impact on selected product features.

Correlation of Smac/DIABLO protein expression with the clinico-pathological features of breast cancer patients.

Smac/DIABLO protein promotes caspase-dependent apoptosis by inhibition of inhibitor of apoptosis protein (IAP) family members. The role of Smac/DIABLO in breast cancer has not been yet established. Therefore, the aim of the study was to assess the expression of this protein in tumor cells from breast cancer patients. The expression of Smac/DIABLO was analyzed in 62 breast cancer patients by flow cytometry. The obtained results were compared with expression of this protein in benign breast tumor tissue, which served as the control (11 patients with fibroadenoma). Expression of caspase-3 proteins in breast cancer was also evaluated. Smac/DIABLO expression in breast cancer was correlated with clinical and pathological data. Although the expression of Smac/DIABLO protein was found in all examined samples of both the breast cancer and fibroadenoma patients, the median expression of Smac/Diablo in breast cancer was significantly lower than in the control (39.1% vs. 48.1%; p=0.0047). Smac/DIABLO expression correlated with expression of caspase-3 (p=0.000008). In pT1 breast cancer patients, expression of Smac/DIABLO protein was higher than in those with pT2-3 (p=0.02). Diffuse cancer infiltration significantly correlated with lower expression of Smac/DIABLO protein (p=0.02). Moreover, there was a loose correlation between low expression of Smac/DIABLO protein and cancer embolus in minor blood and lymphatic vessels (p=0.08). Our results indicate that expression of Smac/DIABLO inversely correlates with the tumor stage, which may suggest that this protein may play an important role in the breast cancer development.

Identification of a subdomain of CENP-B that is necessary and sufficient for localization to the human centromere.

We have combined in vivo and in vitro approaches to investigate the function of CENP-B, a major protein of human centromeric heterochromatin. Expression of epitope-tagged deletion derivatives of CENP-B in HeLa cells revealed that a single domain less than 158 residues from the amino terminus of the protein is sufficient to localize CENP-B to centromeres. Centromere localization was abolished if as few as 28 amino acids were removed from the amino terminus of CENP-B. The centromere localization signal of CENP-B can function in an autonomous fashion, relocating a fused bacterial enzyme to centromeres. The centromere localization domain of CENP-B specifically binds in vitro to a subset of alpha-satellite DNA monomers. These results suggest that the primary mechanism for localization of CENP-B to centromeres involves the recognition of a DNA sequence found at centromeres. Analysis of the distribution of this sequence in alpha-satellite DNA suggests that CENP-B binding may have profound effects on chromatin structure at centromeres.

The centromere: hub of chromosomal activities.

Centromeres are the structures that direct eukaryotic chromosome segregation in mitosis and meiosis. There are two major classes of centromeres. Point centromeres, found in the budding yeasts, are compact loci whose constituent proteins are now beginning to yield to biochemical analysis. Regional centromeres, best described in the fission yeast Schizosaccharomyces pombe, encompass many kilobases of DNA and are packaged into heterochromatin. Their associated proteins are as yet poorly understood. In addition to providing the site for microtubule attachment, centromeres also have an important role in checkpoint regulation during mitosis.

Structure of the human centromere at metaphase.

Until recently the centromere was thought to be a relatively homogeneous region of densely packed heterochromatin with a single differentiated domain--the kinetochore--at its surface, representing the point of attachment of the mitotic spindle. We now know that the centromere of higher eukaryotes is composed of several domains that have been identified using antibody probes. Somewhere within the domains are located both the factor(s) that control the disjunction of sister chromatids and the molecular motor responsible for chromosome movement towards the spindle poles.

Determination of telomere length and telomerase activity in cattle infected with bovine leukaemia virus (BLV).

Telomeres are repetitive sequence structures at the ends of chromosomes. They consist of the double stranded DNA repeats followed by the short single stranded DNA. In humans and other verterbrates the telomeric sequence is composed of tandem of TTAGGG repeats. With each cells division telomeres shorten by up to 200 base pairs. Telomerase is an enzyme responsible for continuous cell growth and is repressed in most somatic cells, except proliferating progenitor cells, but in more than 85% of cancer cells telomerase expression is observed. Tumour cells with metastatic potential may demonstrate a high telomerase activity, allowing cells to escape from the inhibition of cell proliferation due to shortened telomeres. Determination of telomerase expression was performed with the use of PCR ELISA in samples isolated from bovine leukaemia virus (BLV) infected cows. Telomerase activity was found in almost all investigated samples. The relative telomerase activity (RTA) was higher in infected cows than in healthy animals and the differences were statistically significant (α=0.05). In blood lymphocytes of BLV-infected cows the mean values of telomerase expression determined in real-time PCR were 3534.12 copies, in the healthy group there were 1010.10 copies and these differences were also statistically significant. For telomere length evaluation the Telomere PNA/FITC FISH and Telomere PNA/FITC FISH for flow cytometry were used. The mean fluorescence intensity of telomere sequences calculated on the surface of interphase nuclei of leukaemic blood lymphocytes was lower than that in the control animals and the difference was statistically significant. The mean length of telomeres in BLV- infected and healthy cows was 31.63 ± 12.62 and 38.4 ± 4.03, (p=0.112), respectively.

Surprising deficiency of CENP-B binding sites in African green monkey alpha-satellite DNA: implications for CENP-B function at centromeres.

Centromeres of mammalian chromosomes are rich in repetitive DNAs that are packaged into specialized nucleoprotein structures called heterochromatin. In humans, the major centromeric repetitive DNA, alpha-satellite DNA, has been extensively sequenced and shown to contain binding sites for CENP-B, an 80-kDa centromeric autoantigen. The present report reveals that African green monkey (AGM) cells, which contain extensive alpha-satellite arrays at centromeres, appear to lack the well-characterized CENP-B binding site (the CENP-B box). We show that AGM cells express a functional CENP-B homolog that binds to the CENP-B box and is recognized by several independent anti-CENP-B antibodies. However, three independent assays fail to reveal CENP-B binding sites in AGM DNA. Methods used include a gel mobility shift competition assay using purified AGM alpha-satellite, a novel kinetic electrophoretic mobility shift assay competition protocol using bulk genomic DNA, and bulk sequencing of 76 AGM alpha-satellite monomers. Immunofluorescence studies reveal the presence of significant levels of CENP-B antigen dispersed diffusely throughout the nuclei of interphase cells. These experiments reveal a paradox. CENP-B is highly conserved among mammals, yet its DNA binding site is conserved in human and mouse genomes but not in the AGM genome. One interpretation of these findings is that the role of CENP-B may be in the maintenance and/or organization of centromeric satellite DNA arrays rather than a more direct involvement in centromere structure.

The role of p73 in hematological malignancies.

The P73 gene is a homologue of the P53 tumor suppressor. Owing to its structural similarity with p53, p73 was originally considered to have tumor suppressor function. However, the discovery of N-terminal truncated isoforms with oncogenic properties showed a 'two in one' structure of its product, p73 protein. The full-length variants are strong inducers of apoptosis, whereas the truncated isoforms inhibit proapoptotic activity of p53 and the full-length p73. Thus, p73 is involved in the regulation of cell cycle, cell death and development. Moreover, it plays a role in carcinogenesis and controls tumor sensitivity to treatment. p73 is commonly expressed in tumor cells in hematological malignancies. Overexpression of p73 protein and aberrant expression of its particular isoforms, with very low frequency of P73 hypermethylation or mutations, were found in malignant myeloproliferations, including acute myeloblastic leukemia. In contrast, hypermethylation and subsequent inactivation of the P73 gene are the most common findings in malignant lymphoproliferative disorders, especially acute lymphoblastic leukemia (ALL) and non-Hodgkin's lymphomas. Assessment of P73 methylation may provide important prognostic information, as was confirmed in patients with ALL. This review summarizes some aspects of p73 biology with particular reference to its possible pathogenetic role and prognostic significance in hematological malignancies.

Selected miRNA levels are associated with IKZF1 microdeletions in pediatric acute lymphoblastic leukemia.

The clinical outcome of children with high-risk relapsed B-cell precursor acute lymphoblastic leukemia (BCP-ALL) is poor. The present study assessed the utility and prognostic value of selected microRNA (miRNA/miR) in BCP-ALL. The changes in the expression levels of these miRNAs regarding known gene lesions affecting lymphoid development [early B-cell factor 1 (EBF1), ETS variant 6 (ETV6), IKAROS family zinc finger 1 (IKZF1), paired box 5 (PAX5), cyclin dependent kinase inhibitor (CDKN) 2A/CDKN2B, retinoblastoma 1 (RB1), pseudoautosomal region 1 (PAR1), B-cell translocation gene 1 protein (BTG1)] were analyzed. The following miRNAs were analyzed: miR-24, miR-31, miR-128, miR-542, and miR-708. The present study focused on patients with deletions of the IKAROS transcriptional factor gene IKZF1, which is currently considered to be an independent negative prognostic factor for ALL outcome. It was demonstrated that the expression level of miR-128 was significantly lower in patients with IKZF1 deletion compared with patients without IKZF1 deletion. Additionally, low expression of miR-542 was associated with CDKN2A/B and miR-31deletions, and low expression of miR-24 was associated with miR-31 deletion. Low expression of miR-31, miR-24, miR-708 and miR-128 was associated with PAX5 deletion, high expression of miR-24 and miR-542 was associated with PAR1 deletion and high expression of miR-708 was associated with ETV6 deletion. The expression of the selected miRNAs was not associated with deletions of BTG1, EBF1 and RB1. These data, by emphasizing the association of miRNAs expression level with microdeletions, may assist to elucidate ALL biology and contribute to future studies on the possible applications of the miRNA profile for diagnosis.

Addition of cladribine to daunorubicin and cytarabine increases complete remission rate after a single course of induction treatment in acute myeloid leukemia. Multicenter, phase III study.

To assess the efficacy of an original DAC-7 regimen: daunorubicine (DNR) 60 mg/m2/day, days 1-3; cytarabine (AraC) 200 mg/m2/day, days 1-7; cladribine (2-CdA) 5 mg/m2/day, days 1-5, 400 untreated adult acute myeloid leukemia patients (including 63 with preceding myelodysplastic syndrome), aged 45 (16-60) years were randomized to either DAC-7 (n=200) or DA-7 (without 2-CdA, n=200). The overall CR rate equaled 72% for DAC-7 and 69% for DA-7 arm (P=NS). After a single course of DAC-7 induction, the CR rate equaled 64% and was significantly higher compared to 47% in the DA-7 arm (P=0.0009). Median hospitalization time during the induction was 7 days shorter for DAC-7 compared to the DA-7 group (33 vs 40 days, P=0.002). Toxicity was comparable in both groups. The probability of 3-year leukemia-free survival (LFS) for DAC-7 and DA-7 group equaled 43 and 34%, respectively (P=NS). There was a trend toward higher LFS rate for patients aged >40 years receiving DAC-7 compared with DA-7 regimen (44 vs 28%, P=0.05). This study proves that addition of 2-CdA increases antileukemic potency of DNR+AraC regimen, thus resulting in a higher CR rate after one induction cycle when compared to DA-7, without additional toxicity. It shortens hospitalization time and may improve long-term survival in patients aged >40 years.

A thematic framework of illness narratives produced by stroke patients.

PURPOSE: The present study aims at elucidating the impact of stroke on psychosocial functioning of stroke survivors. METHODS: Data were investigated using interpretative thematic analysis of illness stories produced by 29 patients. RESULTS: Eight themes emerged from the data: Medical Information; Interpersonal Changes; Cognitive, Physical and Emotional Functioning; Strategies of Coping; Social Support; and Information Irrelevant to the Question. The most frequent organization of the themes followed the course of medical intervention and rehabilitation. Narrations of individual patients varied in terms of the presence of particular themes, the amount of information on each topic and organization. CONCLUSIONS: The results suggest that the analysis of non-guided illness narratives can be effectively used to identify the thematic areas important to individual stroke patients. The thematic content analysis of stroke stories can allow health professionals to better understand the patient's state of knowledge related to illness as well as his or her socio-psychological functioning which may be useful in the course of planning further assessment and rehabilitation of patients with stroke. Implications for Rehabilitation Experience of illness and life changes following stroke should be recognized as central to the provision of targeted rehabilitation. To understand the subjective perspective a content analysis of the content narrative is recommended. Our study highlights seven general thematic categories that may be regarded as key. The categories may be useful for clinicians to help individuals to clarify their main concerns following a stroke.

Effects of ionizing radiation and beta-adrenergic stimulation on the expression of early response genes in rat parotid glands.

There is little known about the regulation of gene expression in rat parotid glands after exposure to ionizing radiation. The present studies investigate the effects of in vivo ionizing radiation, with subsequent stimulation of beta-adrenergic receptors by isoproterenol, on parotid gland function and on the expression of the early response genes, c-fos, c-jun, and jun B. Ionizing radiation diminished parotid gland weight and saliva output. Treatment of irradiated rats with isoproterenol increased the gland weight to levels similar to those in nonirradiated rats. However, such treatment had no effect on saliva output as indicated by measurements of parotid salivary flow rate. Irradiation alone increased the expression of c-fos, c-jun, and jun B. The combination of irradiation and isoproterenol had an additional effect on the levels of c-fos and jun B mRNAs and proteins particularly at earlier experimental times (1 to 8 h). Isoproterenol alone induced high levels of c-fos and jun B mRNA but not of c-jun mRNA. However, c-jun mRNA was induced markedly by radiation and 8 h of isoproterenol treatment, indicating a combined effect on c-jun gene expression. These observations suggest that the expression of the proto-oncogenes c-fos, c-jun, and jun B is probably regulated through differential signal transduction pathways which may be activated by these external stimuli and may be associated with functional changes induced in the rat parotid gland by ionizing radiation and by ionizing radiation and isoproterenol.

Immunosuppressive drugs inhibit class II antigen (HLA-D) synthesis by B and T lymphocytes.

The influence of immunosuppressive drugs on class II (HLA-D) antigen synthesis and expression by monocytes and B and T cells was studied. The constitutive HLA-D expression on monocytes and malignant B cells was resistant, while the acquisition of those markers by stimulated T cells very sensitive to immunosuppression. Active derivative of cyclophosphamide (4HCy) and cyclosporine proved to be most efficacious inhibitors of class II antigens.

Large granular lymphocyte lymphocytosis with neutropenia: a case report.

A case of large granular lymphocyte lymphocytosis with neutropenia was studied as evidenced by peripheral lymphocytosis of cells having typical morphology and profound neutropenia. Surface markers analysis revealed that almost all cells were CD8+ and their subpopulation DR+. The cells had strong spontaneous and inducible suppressor functions in vitro. Moreover, their ADCC activity was strong but NK activity very weak.

Ciprofloxacin prophylaxis for patients undergoing high-dose chemotherapy and autologous stem cell transplantation (ASCT) - a single-center experience.

PURPOSE: The objective of the study was to evaluate the efficacy of ciprofloxacin prophylaxis for patients undergoing high-dose chemotherapy followed by autologous stem cell transplantation (ASCT). MATERIALS AND METHODS: The data of 104 patients transplanted at the Department of Hematology Medical University of Lodz between 2005 and 2008 were analyzed. The cohort was divided into two groups depending on the administered ciprofloxacin prophylaxis. Conditioning regimens did not differ significantly among the groups. Multiple myeloma was the main indication for ASCT in both groups. RESULTS: Ciprofloxacin prophylaxis resulted in a statistically significant reduction of duration of intravenous (IV) antibiotic treatment in the group with prophylaxis (p=0.01). The trend has been observed towards lower prevalence of infectious episodes in the prophylaxis group. Positive blood cultures were similar in both groups with no significant resistance to ciprofloxacin. CONCLUSION: These data demonstrate that ciprofloxacin prophylaxis is beneficial for patients treated with ASCT following high dose chemotherapy regimen, in terms of the intravenous antibiotics usage. This advantage directly translates into economic benefit and may also induce less bacterial resistance due to less exposure to antibiotics.

Novel and emerging drugs for acute myeloid leukemia: pharmacology and therapeutic activity.

For the last twenty years, significant progress in Molecular and Cellular Biology has resulted in a better characterization and understanding of the biology and prognosis of acute myeloid leukemia (AML). These achievements have provided new opportunities for the development of innovative, more effective therapies. Novel agents potentially useful in the treatment of patients with AML include new formulations of established drugs, newer nucleoside analogs, molecular target drugs, monoclonal antibodies and other agents. Three newer nucleoside analogs, clofarabine, troxacitabine and sapacitabine have been recently investigated in patients with AML. Two methylation inhibitors, 5-azacyticline and decitabine are pyrimidine nucleoside analogs of cytidine which can be incorporated into RNA and/or DNA. Lower doses of these agents are active in AML and have been extensively investigated, especially in secondary AML and AML in elderly patients. Tipifarnib and lonafarnib are orally available farnesyltransferase inhibitors with in vitro and in vivo activity against AML. In recent years, FLT3 inhibitors, lestaurinib, tandutinib and PKC 412 have been developed and tested in AML. The preclinical observations and clinical studies indicate that FLT3 inhibitors are promising agents in the treatment of FLT3 mutated AML patients, especially when used in combinations with chemotherapy. Several newer MDR inhibitors, including valspodar (PSC-833) and zosuquidar trihydrochloride have been also tested for the treatment of relapsed AML. This article reviews the various classes of AML targets and drugs that are under early phase clinical evaluation, especially those that are likely to enter clinical practice in the near future.